WO2018018010A1 - Fgf mutants and uses thereof - Google Patents

Fgf mutants and uses thereof Download PDF

Info

Publication number
WO2018018010A1
WO2018018010A1 PCT/US2017/043383 US2017043383W WO2018018010A1 WO 2018018010 A1 WO2018018010 A1 WO 2018018010A1 US 2017043383 W US2017043383 W US 2017043383W WO 2018018010 A1 WO2018018010 A1 WO 2018018010A1
Authority
WO
WIPO (PCT)
Prior art keywords
modification
polypeptide
amino acid
fgf
isolated
Prior art date
Application number
PCT/US2017/043383
Other languages
French (fr)
Inventor
Mack Flinspach
Ross FELLOWS
Andrew PIEKARZ
Matt LUNDBERG
James MARION
Original Assignee
Metacrine, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Metacrine, Inc. filed Critical Metacrine, Inc.
Publication of WO2018018010A1 publication Critical patent/WO2018018010A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C07K14/50—Fibroblast growth factors [FGF]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/427—Thiazoles not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • Metabolic diseases or conditions such as diabetes and obesity are growing health problems worldwide. In some instances, it is estimated that more than 250 million adults worldwide have diabetes and the figure is projected to increase to 380 million by 2025.
  • FGF mutants include FGF mutants and their use in the treatment of one or more metabolic diseases or conditions.
  • the FGF mutant is an FGF1 mutant.
  • an FGF1 mutant is used for the treatment of one or more metabolic diseases or conditions.
  • an isolated and purified FGF polypeptide comprising at least 80% sequence identity to SEQ ID NOs: 3-25917 and 25922-25974.
  • the FGF polypeptide comprises at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NOs: 3-25917 and 25922-25974.
  • the FGF polypeptide consists of SEQ ID NOs: 3-25917 and 25922-25974.
  • the FGF polypeptide comprises at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NOs: 130, 131, 132, 133, 134, 135, 139, 141, 142, 143, 144, 146, 147, 148, 149, 150, 151, 152, 153, 156, 157, 158, 159, 163, 165, 168, 170, 171, 174, 177, 284, 286, 290, 401, 484, 488, or 518.
  • the FGF polypeptide further comprises a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2.
  • the FGF polypeptide further comprises a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L89, T96, K128, or a combination thereof, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2.
  • the FGF polypeptide further comprises a modification at amino acid residue L84, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2.
  • the modification comprises a mutation to a non-polar residue or a charged residue.
  • the modification comprises a mutation to alanine.
  • the isolated and purified FGF polypeptide comprises a modification of T30A, T34A, A48G, E53A, K57A, K57E, Q63A, T78A, P79A, L84A, L89A, T96A, K128E, or a combination thereof.
  • the isolated and purified FGF polypeptide has enhanced stability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild- type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide.
  • the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide is an FGF1 mutant.
  • the FGF polypeptide comprises a modification at amino acid residue L84, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2.
  • the isolated and purified FGF polypeptide further comprises a modification at amino acid residue H41, 142, or Q43, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2.
  • the isolated and purified FGF polypeptide further comprises a modification at amino acid residue K10, Y15, S17, N18, H21, L26, P27, D28, R35, D36, R37, S38, D39, Q40, Q43, S47, Y55, E60, T61, Y64, D68, E81, E87, R88, E90, E91, N92, Y94, N95, K100, K101, H102, K105, Kl 13, S I 16, CI 17, Kl 18, P121, R122, H124, Q127, or L133, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2.
  • the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild- type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide is an FGF1 mutant.
  • an isolated and purified FGF polypeptide comprising: one or more modified amino acids selected from Group 1 amino acid residues: P27, R35, D36, R37, Y55, K57, T61, E81, E87, R88, N92, T96, K101, H102, and L133; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; wherein the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • an isolated and purified FGF polypeptide comprising two or more modifications, the polypeptide comprising: a first modification at an amino acid residue selected from: Y15, R35, R37, K57, R88, N92, T96, R122, and L133; and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S 17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, SI 16, Rl 19, H124, Y125, P136, V137, S138, S139, and D140; wherein the numbering of the amino acid residues corresponds to S
  • the first modification is at R88, and the second modification is at an amino acid residue selected from the group consisting of E91 and T96.
  • the first modification is at K57, and the second modification is at an amino acid residue selected from the group consisting of: K10, A48, E90, and E91.
  • the first modification is at K57, and the second modification is at an amino acid residue selected from the group consisting of: K10, A48, and E90.
  • the first modification is at T96, and the second modification is at Q43. In some embodiments, the first modification is at Y 15, and the second modification is at Q43.
  • the first modification is at R122
  • the second modification is at an amino acid residue selected from the group consisting of: R35, R37, and K57.
  • the FGF polypeptide further comprises a truncation.
  • the truncation is an N-terminal truncation.
  • the N-terminal truncation is a deletion of amino acid residues 1-9.
  • the modification comprises a mutation to a non- polar residue or a charged residue.
  • the modification comprises a mutation to alanine.
  • K57 is mutated to an amino acid selected from the group consisting of: alanine (A), asparagine (N), glutamic acid (E), and histidine (H).
  • R88 is mutated to leucine (L).
  • N92 is mutated to an amino acid selected from the group consisting of: alanine (A) and threonine (T).
  • T96 is mutated to alanine (A).
  • the FGF polypeptide is an FGFl polypeptide.
  • the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide.
  • the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide.
  • the modification comprises a mutation to a non-polar residue or a charged residue. In some embodiments, the modification comprises a mutation to alanine.
  • the FGF polypeptide is an FGFl polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide.
  • the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide.
  • an isolated and purified FGF polypeptide comprising at least 80% sequence identity to a sequence selected from SEQ ID NOs: 3-25917 and 25922-25974.
  • the FGF polypeptide comprises at least 85%, 90%, 95%, 96%), 97%), 98%o, 99%o, or more sequence identity to a sequence selected from SEQ ID NOs: 3- 25917 and 25922-25974.
  • the FGF polypeptide consists of a sequence selected from SEQ ID NOs: 3-25917 and 25922-25974.
  • the FGF polypeptide is an FGFl polypeptide.
  • the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide.
  • an isolated and purified FGF polypeptide having an FGFR interaction activity that is equivalent to the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2; and a mitogenicity that is at least 1-fold lower than the mitogenicity of the FGFl polypeptide of SEQ ID NO: 2.
  • the FGFR interaction activity of the isolated and purified FGF polypeptide is about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% to the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2.
  • the mitogenicity is at least 2-fold lower, 3 -fold lower, 4-fold lower, 5 -fold lower, 6-fold lower, 7-fold lower, 8-fold lower, 9-fold lower, or 10-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2.
  • the isolated and purified FGF polypeptide is an isolated and purified FGF polypeptide described supra.
  • the FGF polypeptide is an FGF1 polypeptide.
  • the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide.
  • the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild-type FGF polypeptide.
  • the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide.
  • the at least one additional FGF polypeptide comprises at least one modification, a truncation, or a combination thereof.
  • the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
  • the pharmaceutical composition is formulated for oral or parenteral administration.
  • a method of treating a metabolic disease or condition in a subject in need thereof comprising administering to the subject a therapeutically effective amount of an isolated and purified FGF polypeptide described above.
  • the metabolic disease or condition comprises FGF 1 -related disease or condition.
  • the metabolic disease or condition comprises diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), dyslipidemia, a cardiovascular disease, Hurler syndrome, Huntington's disease, Alzheimer's disease, Parkinson's disease, Crohn's disease, Fabry disease, or galactosemia.
  • the metabolic disease or condition comprises impaired insulin sensitivity or glucose intolerance.
  • the subject has diabetes.
  • the diabetes comprises type 1 diabetes, type 2 diabetes, type 3 diabetes, type 4 diabetes, double diabetes, latent autoimmune diabetes (LAD), gestational diabetes, neonatal diabetes mellitus (NDM), maturity onset diabetes of the young
  • the subject has type 1 diabetes. In some embodiments, the subject has type 2 diabetes. In some embodiments, the subject has type 4 diabetes. In some embodiments, the subject has prediabetes. In some embodiments, the subject has polycystic ovary syndrome. In some embodiments, the subject has metabolic syndrome. In some embodiments, the subject has obesity. In some embodiments, the subject has non-alcoholic fatty liver disease. In some embodiments, the subject has insulin resistance. In some embodiments, the subject has lipodystrophy. In some embodiments, the subject has a cardiovascular disease.
  • the cardiovascular disease comprises hypertension, coronary artery diseases (CAD), stroke, rheumatic heart disease, hypertensive heart disease, cardiomyopathy, heart arrhythmia, congenital heart disease, carditis, aortic aneurysms, renal artery stenosis, peripheral artery disease, venous thrombosis, heart valve disease, deep vein thrombosis, cerebrovascular disease, cardiac dysrhythmia, chronic obstructive pulmonary disease, or inflammatory heart disease.
  • the subject has hypertension.
  • the method further comprises administration of an additional therapeutic agent.
  • the additional therapeutic agent comprises thiazolidinedione.
  • thiazolidinedione comprises pioglitazone, rosiglitazone, lobeglitazone, and troglitazone.
  • the FGF polypeptide and the additional therapeutic agent are administered
  • kits comprising an isolated and purified FGF polypeptide described above.
  • kits comprising a polynucleic acid molecule encoding an FGF polypeptide described above.
  • Fig. lA-Fig. IB illustrate the location of the FGF1 Group 1 residues with respect to a FGFl-FGFRlc complex (PDB: 30JV.pdb) in a first orientation.
  • Fig. 1A shows the FGFl-FGFRlc complex and
  • Fig. IB shows a close-up of the positions of the Group 1A, Group IB, and Group 1C residues on FGF1.
  • Fig. 2A-Fig. 2B illustrate the location of the FGF1 Group 1 residues with respect to a FGFl-FGFRlc complex (PDB: 30JV.pdb) in a second orientation.
  • Fig. 2A shows the FGFl-FGFRlc complex and
  • Fig. 2B shows a close-up of the positions of the Group 1A, Group IB, and Group 1C residues on FGF1.
  • Fig. 3A-Fig. 3B illustrate the location of the FGF1 Group 1 and Group 2 residues with respect to a FGFl-FGFRlc complex (PDB: 30JV.pdb) in a first orientation.
  • Fig. 3A shows the FGFl-FGFRlc complex and
  • Fig. 3B shows a close-up of the positions of the Group 1 and Group 2 residues on FGF1.
  • Fig. 4A-Fig. 4B illustrate the location of the FGF1 Group 1 and Group 2 residues with respect to a FGFl-FGFRlc complex (PDB: 30JV.pdb) in a second orientation.
  • Fig. 4A shows the FGFl-FGFRlc complex and
  • Fig. 4B shows a close-up of the positions of the Group 1 and Group 2 residues on FGF1.
  • Fig. 6 illustrates the mitogenic activity of exemplary FGF1 polypeptides in zona glomerulosa.
  • Fig. 7 illustrates the mitogenic activity of exemplary FGF1 polypeptides in zona fasciculata/reticularis.
  • Fig. 9 illustrates the mitogenic activity of exemplary FGF1 polypeptides in hepatocytes.
  • Fig. 13A-Fig. 13C illustrate in-vivo mitogenicity and in-vivo efficacy of exemplary FGF1 polypeptides.
  • Fig. 13A shows the in-vivo mitogenicity of exemplary FGF1 polypeptides.
  • Fig. 13B and Fig. 13C show the in-vivo efficacy of exemplary FGF1 polypeptides.
  • Fibroblast growth factors are a family of growth factors that are involved in endocrine signaling pathways, wound healing, angiogenesis, and embryonic development. There are 22 structurally related members in the FGF family, which are further classified as intracrine, paracrine, and endocrine FGFs, based on their action mechanism. Paracrine and endocrine FGFs are extracellular signaling molecules that interact with fibroblast growth factor receptors (FGFRs).
  • FGFRs fibroblast growth factor receptors
  • FGF19/FGF15 FGF 15 is the mouse homolog of human FGF 19
  • FGF21 FGF23
  • Intracrine FGFs are FGFR-independent intracellular molecules that regulate the function of voltage gated sodium channels and the members include FGF11, FGF12, FGF13, and FGF14.
  • FGF1 interacts with all four FGFRs, fibroblast growth factor receptor 1 (FGFR1), fibroblast growth factor receptor 2 (FGFR2), fibroblast growth factor receptor 3 (FGFR3), and fibroblast growth factor receptor 4 (FGFR4), and in particular, isoforms FGFRlb (FGFRlIIIb), FGFRlc (FGFRlIIIc), FGFR2b (FGFR2IIIb), FGFR2c (FGFR2IIIc), FGFR3b (FGFR3IIIb), FGFR3c (FGFR3IIIc), and FGFR4.
  • FGFRlb fibroblast growth factor receptor 1
  • FGFR2b fibroblast growth factor receptor 2
  • FGFR3 fibroblast growth factor receptor 3
  • FGFR4 fibroblast growth factor receptor 4
  • the full length FGF1 (GenBank Accession Nos: BC032697.1 and AAH32697.1) comprises a non-glycosylated 155 amino acid residue polypeptide (SEQ ID NO: 1).
  • the mature FGF1 comprises 140 amino acid residues in length (SEQ ID NO: 2), which is resulted from the truncation of the first 15 residues from the N-terminus of the full length FGF1.
  • FGF1 is expressed in metabolic tissues such as liver, kidney, muscle, and white adipose tissue (WAT).
  • WAT white adipose tissue
  • FGF1 is upregulated in WAT following a high-fat food intake.
  • Each cell of the white adipose tissue, or white fat comprises a single large droplet of fat with a nucleus encircling the droplet as a thin rim.
  • WAT is further grouped into visceral or gonadal WAT (gWAT) or inguinal white adipose tissue (iWAT).
  • gWAT visceral or gonadal WAT
  • iWAT inguinal white adipose tissue
  • FGF1 has been observed in the adipocyte fraction but not in the stromal vascular fraction (SVF) of gWAT and in the adipocyte fraction of iWAT.
  • SVF stromal vascular fraction
  • FGF1 knockout mice has been characterized by aberrant WAT expansion, impaired WAT vascularization, and rapid development of severe diabetes upon high-fat diet (HFD) challenge and defective WAT reduction upon HFD withdrawal (Jonker, et al, "A PPARy-FGFl axis is required for adaptive adipose remodeling and metabolic homeostasis," Nature 485(7398): 391-394 (2012)).
  • Fibroblast growth factor 2 also known as basic fibroblast growth factor, bFGF, FGF2, or FGF- ⁇
  • FGF2 is a 288 amino acid polypeptide that comprises about 5 different isoforms. The C-terminal portion of FGF2 shares about 48% sequence identity to FGF 1. Similar to FGF 1 , FGF2 is involved in cellular developmental processes such as proliferation, migration, differentiation, and survival. In some instances, FGF2 has been implicated in the pathogenesis of vascular dysfunction induced by hyperglycemia, such as vascular
  • the mitogenic activity of FGF 1 is potentiated by heparan sulfate or a heparan sulfate analogue, leading to cellular developments such as proliferation, migration, and differentiation.
  • the mitogenic activity of FGF 1 has also been linked to increased tumorigenicity. Indeed, studies have shown that FGF1 stimulate the cell growth of breast cancer, prostate cancer, and bladder cancer. See, e.g., Fernig, et al., "Differential regulation of FGF-1 and -2 mitogenic activity is related to their kinetics of binding to heparan sulfate in MDA-MB-231 human breast cancer cells," Biochemical and Biophysical Research Comm. 267(3): 770-776 (2000);
  • FGF mutants and methods of use in the treatment of a metabolic disease exhibit low mitogenicity. In some cases, the FGF mutants further exhibit near wild-type FGFR interaction. In additional cases, the FGF mutants further induces and/or enhances insulin sensitivity in a subject. In some instances, the FGF mutant is an FGF1 mutant. In some instances, described herein are FGF1 mutants and methods of use in the treatment of a metabolic disease. In some cases, the FGF1 mutants exhibit low mitogenicity and near wild-type FGFR interaction, and further optionally induces and/or enhances insulin sensitivity in a subject.
  • an isolated and purified FGF polypeptide such as an isolated and purified FGF polypeptide which comprises at least 80% sequence identity to SEQ ID NOs: 3-25917 and 25922- 25974; an isolated and purified FGF polypeptide comprising a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof; or an isolated and purified FGF polypeptide that comprises a modification at amino acid residue K10, Y15, S 17, N18, H21, L26, P27, D28, R35, D36, R37, S38, D39, Q40, Q43, S47, Y55, E60, T61, Y64, D68, E81, E87, R88, E90, E91
  • FGF mutants and methods of use in the treatment of a metabolic disease.
  • the FGF mutant is an FGFl or FGF2 mutant.
  • the FGF mutant is an FGF 1 mutant.
  • the FGF mutant is an FGFl polypeptide that comprises 155 amino acid residues (or comprises the full length FGFl sequence; SEQ ID NO: 1). In some instances, the FGF mutant is an FGFl polypeptide that comprises 140 amino acid residues (or comprises the mature FGFl sequence; SEQ ID NO: 2). In some instances, the FGF mutant is a mature FGFl (140 amino acid residues) that further comprises a truncation at the N-terminus, the C-terminus, or truncations at both termini. In some instances, the truncation is a truncation of about 1 to about 30 amino acid residues.
  • the FGF mutant is an FGFl polypeptide that comprises about 14 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 15 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 16 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 17 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 18 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 19 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 20 modifications. In some cases, a modification comprises an insertion, a deletion, or a mutation.
  • the mutation is a missense mutation or a nonsense mutation.
  • the insertion comprises a duplication or one or more amino acid residues or a repeat expansion of one or more amino acid residues.
  • the FGF mutant is a truncated FGF 1 polypeptide that further comprises one or more modifications described herein.
  • the FGF mutant is a N- terminus truncated FGFl polypeptide that further comprises about 1 to about 20 modifications.
  • the FGF mutant is a C-terminus truncated FGFl polypeptide that further comprises about 1 to about 20 modifications.
  • dicyclohexyl ammonium salt L-cyclohexylglycine, L- phenylglycine, L-propargylglycine, L-norvaline, N-a-aminomethyl-L-alanine, D-a,y-diaminobutyric acid, L-a,y-diaminobutyric acid, ⁇ -cyclopropyl-L-alanine, ( ⁇ -(2,4-dinitrophenyl))-L-a ⁇ - diaminopropionic acid, ( ⁇ - ⁇ - 1 -(4,4-dimethyl-2,6-dioxocyclohex- 1 -ylidene)ethyl)-D-a ⁇ - diaminopropionic acid, ( ⁇ - ⁇ - 1 -(4,4-dimethyl-2,6-dioxocyclohex- 1 -ylidene)ethyl)-L-a ⁇ - diaminopropi
  • Exemplary amino acid analogs of aspartic and glutamic acids include, but are not limited to, a-methyl-D-aspartic acid, a-methyl-glutamic acid, a-methyl-L-aspartic acid, ⁇ -methylene-glutamic acid, (N-Y-ethyl)-L-glutamine, [N-a-(4-aminobenzoyl)]-L-glutamic acid, 2,6-diaminopimelic acid, L- a-aminosuberic acid, D-2-aminoadipic acid, D-a-aminosuberic acid, a-aminopimelic acid, iminodiacetic acid, L-2-aminoadipic acid, threo- -methyl-aspartic acid, ⁇ -carboxy-D-glutamic acid ⁇ , ⁇ -di-t-butyl ester, ⁇ -carboxy-L-glutamic acid ⁇ , ⁇ -di-t-
  • the modification comprises a mutation to a charged amino acid residue.
  • charged amino acids include lysine, arginine, histidine, aspartate, glutamate, or analog thereof.
  • a FGF mutant described herein comprises a mutation to a charged amino acid residue (e.g., a mutation to lysine, arginine, histidine, aspartate, glutamate, or analog thereof).
  • the FGF mutant is an FGFl polypeptide that comprises a modification at at least one amino acid position: 30, 34, 41, 42, 43, 48, 53, 57, 63, 78, 79, 84, 89, 96, or 128, according to the amino acid residue position of SEQ ID NO: 2.
  • the FGF mutant is an FGFl polypeptide that comprises a modification at at least one amino acid position: 30, 34, 48, 53, 57, 63, 78, 79, 84, 89, 96, or 128, according to the amino acid residue position of SEQ ID NO: 2.
  • the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 34, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 41, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 42, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 43, according to the amino acid residue position of SEQ ID NO: 2.
  • the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 48, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 53, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 57, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 63, according to the amino acid residue position of SEQ ID NO: 2.
  • the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 78, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 79, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 84, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 89, according to the amino acid residue position of SEQ ID NO: 2.
  • the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 96, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 128, according to the amino acid residue position of SEQ ID NO: 2.
  • the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 26, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 27, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 28, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid positions 35, according to the amino acid residue position of SEQ ID NO: 2.
  • the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 64, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 68, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 81, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 87, according to the amino acid residue position of SEQ ID NO: 2.
  • the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 93, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 94, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 95, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 100, according to the amino acid residue position of SEQ ID NO: 2.
  • the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 101, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 102, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 105, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 108, according to the amino acid residue position of SEQ ID NO: 2.
  • the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 133, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 134, according to the amino acid residue position of SEQ ID NO: 2.
  • an isolated and purified FGF polypeptide described herein comprises a modification at amino acid residue T30, T34, H41, 142, Q43, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof, according to the amino acid residue numbering of SEQ ID NO: 2.
  • an isolated and purified FGF polypeptide described herein comprises a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof, according to the amino acid residue numbering of SEQ ID NO: 2.
  • the isolated and purified FGF polypeptide comprising a modification at amino acid residue T30, T34, H41, 142, Q43, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof further comprises at least one additional modification.
  • the at least one additional modification comprises an insertion, a deletion, or a mutation.
  • the mutation comprises a mutation to a polar residue, a non-polar residue, or a charged residue.
  • the isolated and purified FGF polypeptide further comprises a truncation.
  • the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
  • the isolated and purified FGF polypeptide comprises a modification of T30A, T34A, A48G, E53A, K57A, K57E, Q63A, T78A, P79A, L84A, L89A, T96A, K128E, or a combination thereof.
  • the isolated and purified FGF polypeptide comprises T30A modification.
  • the isolated and purified FGF polypeptide comprises T34A modification.
  • the isolated and purified FGF polypeptide comprises A48G modification.
  • the isolated and purified FGF polypeptide comprises E53A modification.
  • the isolated and purified FGF polypeptide comprises K57A modification.
  • the mutation comprises a mutation to a polar residue, a non-polar residue, or a charged residue.
  • the isolated and purified FGF polypeptide further comprises a truncation.
  • the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
  • the mutation comprises a mutation to a polar residue, a non- polar residue, or a charged residue.
  • the isolated and purified FGF polypeptide further comprises a truncation.
  • the truncation is an N-terminal truncation, a C- terminal truncation, or a combination thereof.
  • an isolated and purified FGF polypeptide described herein comprises a modification at amino acid residue Kl 18, according to the amino acid residue numbering of SEQ ID NO: 2.
  • the modification comprises a mutation to a polar residue.
  • the modification comprises a mutation to a non-polar residue.
  • a FGF polypeptide comprising one or more modified amino acids at Group 1 amino acid positions: 27, 35, 36, 37, 55, 57, 61, 81, 87, 88, 92, 96, 101, 102, and 133; and one or more modified amino acids at Group 2 amino acid positions: 26, 30, 39, 43, 64, 68, 77, 80, 82, 84, 86, 105, 120, 127, and 128, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the FGF polypeptide comprises one or more modified amino acids at Group 1C amino acid positions: 27, 55, 57, 61, 81, 101, and 102; and one or more modified amino acids at Group 2 amino acid positions: 26, 30, 39, 43, 64, 68, 77, 80, 82, 84, 86, 105, 120, 127, and 128, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the FGF polypeptide is an FGF1 polypeptide.
  • a FGF polypeptide described herein comprises one or more modified amino acids selected from Group 1 amino acid residues: P27, R35, D36, R37, Y55, K57, T61, E81, E87, R88, N92, T96, K101, H102, and L133; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the Group 1 modified amino acids are further divided into Group 1A amino acids: R35, D36, and R37; Group IB amino acids: E87, R88, N92, T96, and L133; and Group IC amino acids: P27, Y55, K57, T61, E81, K101, and H102.
  • the FGF polypeptide comprises one or more modified amino acids selected from Group 1A amino acid residues: R35, D36, and R37; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the FGF polypeptide comprises a Group 1 modified amino acid at T96 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • Group 1 modified amino acids are further divided into Group 1A amino acids: R35, D36, and R37; Group IB amino acids: E87, R88, N92, T96, and L133; and Group IC amino acids: P27, Y55, K57, T61, E81, K101, and HI 02.
  • the FGF polypeptide comprises a first modification at R35, and a second modification at S38; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at Q40; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at D 140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at E81; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • R122 is mutated to an amino acid selected from: : aspartic acid (D), phenylalanine (F), glycine (G), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), glutamine (Q), serine (S), valine (V), and tyrosine (Y).
  • R122 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F).
  • R122 is mutated to glutamic acid (E).
  • the first modification is Y15.
  • the FGFl polypeptide comprises a first modification at Y15, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the FGFl polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: A48, E91, N92, H93, 198, K101, and V137; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the FGFl polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: G6, K9, and S I 7; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the FGFl polypeptide comprises a first modification at R35, and a second modification at E81; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the FGFl polypeptide comprises a first modification at R35, and a second modification at R37; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • R35 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F).
  • R35 is mutated to alanine (A) or glutamic acid (E).
  • the first modification is K57.
  • the FGF1 polypeptide comprises a first modification at K57, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S 138, S 139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • alanine A
  • aspartic acid D
  • glutamic acid E
  • phenylalanine F
  • G G
  • histidine H
  • isoleucine I
  • lysine K
  • leucine L
  • methionine M
  • asparagine N
  • glutamine Q
  • S serine
  • V valine
  • Y tyrosine
  • R122 is mutated to an amino acid selected from: : aspartic acid (D), phenylalanine (F), glycine (G), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), glutamine (Q), serine (S), valine (V), and tyrosine (Y).
  • R122 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F).
  • R122 is mutated to glutamic acid (E).
  • R122 is mutated to alanine (A). In some cases, R122 is mutated to aspartic acid (D). In some cases, R122 is mutated to phenylalanine (F). In some cases, R122 is mutated to glycine (G). In some cases, R122 is mutated to histidine (H). In some cases, R122 is mutated to isoleucine (I). In some cases, R122 is mutated to lysine (K). In some cases, R122 is mutated to leucine (L). In some cases, R122 is mutated to methionine (M). In some cases, R122 is mutated to asparagine (N).
  • the FGF polypeptide comprises a modification at amino acid position 72. In some cases, the FGF polypeptide comprises a modification at amino acid position 76. In some cases, the FGF polypeptide comprises a modification at amino acid position 77. In some cases, the FGF polypeptide comprises a modification at amino acid position 78. In some cases, the FGF polypeptide comprises a modification at amino acid position 79. In some cases, the FGF polypeptide comprises a modification at amino acid position 96. In some cases, the FGF polypeptide comprises a modification at amino acid position 98. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine.
  • the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N- terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
  • the FGF polypeptide comprises a first modification at R35 and a second modification at E53. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Y55. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at K57. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at E60. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at T61. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Q63. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at D68.
  • the FGFl polypeptide comprises a first modification at R35 and a second modification at H102. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at K 105. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at N 106. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at F 108. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Kl 12. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Kl 13.
  • a FGF polypeptide comprising a first modification at amino acid position 133 and a second modification at an amino acid position selected from: 34, 37, 43, 84, 88, 95, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the modification comprises a mutation to a non-polar residue or a charged residue.
  • the modification comprises a mutation to alanine.
  • the FGF polypeptide further comprises a truncation.
  • the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
  • the truncation is an N- terminal truncation.
  • the N-terminal truncation comprises a deletion of amino acid residues 1-9.
  • the FGF polypeptide comprises a first modification at K10 and a second modification at SI 16. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non- polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N- terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
  • the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
  • described herein is a FGF polypeptide comprising a first modification at S I 16 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the FGF polypeptide comprises a first modification at S I 16 and a second modification at Y 15.
  • the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
  • the FGF polypeptide comprises a first modification at R122 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at K101.
  • the FGF polypeptide comprises a first modification at R122 and a second modification at Kl 18.
  • the modification comprises a mutation to a non-polar residue or a charged residue.
  • the modification comprises a mutation to alanine.
  • the FGF polypeptide further comprises a truncation.
  • the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
  • the truncation is an N- terminal truncation.
  • the N-terminal truncation comprises a deletion of amino acid residues 1-9.
  • the FGFl polypeptide comprises a first modification at R122 and a second modification at R35. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at D36. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at L84. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at E87. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at R88. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at Y94.
  • the FGF polypeptide comprises a first modification at H124 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at HI 24 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at K101.
  • the FGF polypeptide comprises a first modification at Y64 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at N95.
  • a FGF polypeptide comprising a first modification at amino acid position 53 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the modification comprises a mutation to a non-polar residue or a charged residue.
  • the modification comprises a mutation to alanine.
  • the FGF polypeptide further comprises a truncation.
  • the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
  • the truncation is an N-terminal truncation.
  • the N-terminal truncation comprises a deletion of amino acid residues 1-9.
  • the FGF polypeptide comprises a first modification at E53 and a second modification at Kl 18.
  • the modification comprises a mutation to a non-polar residue or a charged residue.
  • the modification comprises a mutation to alanine.
  • the FGF polypeptide further comprises a truncation.
  • the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
  • the truncation is an N- terminal truncation.
  • the N-terminal truncation comprises a deletion of amino acid residues 1-9.
  • a FGF polypeptide comprising a first modification at amino acid position 47 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the modification comprises a mutation to a non-polar residue or a charged residue.
  • the modification comprises a mutation to alanine.
  • the FGF polypeptide further comprises a truncation.
  • the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
  • the truncation is an N-terminal truncation.
  • the N-terminal truncation comprises a deletion of amino acid residues 1-9.
  • the FGF polypeptide comprises a first modification at S47 and a second modification at Kl 18.
  • the modification comprises a mutation to a non-polar residue or a charged residue.
  • the modification comprises a mutation to alanine.
  • the FGF polypeptide further comprises a truncation.
  • the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
  • the truncation is an N-terminal truncation.
  • the N-terminal truncation comprises a deletion of amino acid residues 1-9.
  • the FGF1 polypeptide comprises a first modification at S47 and a second modification at R35. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at D36. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at L84. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at E87. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at R88. In some instances, the FGF 1 polypeptide comprises a first modification at S47 and a second modification at Y94.
  • the FGF1 polypeptide comprises a first modification at Q40 and a second modification at N95. In some instances, the FGF1 polypeptide comprises a first modification at Q40 and a second modification at K101. In some instances, the FGF1 polypeptide comprises a first modification at Q40 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
  • a FGF polypeptide comprising a first modification at S 17 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
  • the FGF polypeptide comprises a first modification at S 17 and a second modification at Y 15.
  • the FGF polypeptide comprises a first modification at S17 and a second modification at D28.
  • the FGF polypeptide comprises a first modification at S 17 and a second modification at R35.
  • the FGF mutant is an FGFl polypeptide comprising about 94% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 95% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 96% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 97% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
  • the FGF mutant is an FGFl polypeptide comprising about 98% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 99% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 100% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide consists of a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
  • the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 134. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 135.
  • the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 159. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 163.
  • the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 165. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 168.
  • the FGF mutant is an FGF1 polypeptide consists of a polypeptide of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 139, 141, 142, 143, 144, 146, 147, 148, 149, 150, 151, 152, 153, 156, 157, 158, 159, 163, 165, 168, 170, 171, 174, 177, 284, 286, 290, 401, 484, 488, or 518.
  • the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 80% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 85% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 90% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 91% or more sequence identity to SEQ ID NO: 25851.
  • the FGF mutant comprises about 92% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 93% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 94% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 95% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 96% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 97% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 98% or more sequence identity to SEQ ID NO: 25851.
  • the FGF mutant has a FGFR interaction activity that is about 20% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 30% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 40% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 50% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2.
  • FGF polypeptides described herein are generated recombinantly through a host cell system.
  • the host cell is a eukaryotic cell (e.g., mammalian cell, insect cells, yeast cells or plant cell) or a prokaryotic cell (e.g., gram -positive bacterium or a gram-negative bacterium).
  • a eukaryotic host cell is an insect host cell.
  • exemplary insect host cell include Drosophila S2 cells, Sf9 cells, Sf21 cells, High FiveTM cells, and expresSF+® cells.
  • Bacterial vectors include, for example, pACYC177, pASK75, pBAD vector series, pBADM vector series, pET vector series, pETM vector series, pGEX vector series, pHAT, pHAT2, pMal-c2, pMal-p2, pQE vector series, pRSET A, pRSET B, pRSET C, pTrcHis2 series, pZA31-Luc, pZE21-MCS-l, pFLAG ATS, pFLAG CTS, pFLAG MAC, pFLAG Shift- 12c, pTAC-MAT-1, pFLAG CTC, or pTAC-MAT-2.
  • Insect vectors include, for example, pFastBacl, pFastBac DUAL, pFastBac ET, pFastBac HTa, pFastBac HTb, pFastBac HTc, pFastBac M30a, pFastBact M30b, pFastBac, M30c, pVL1392, pVL1393, pVL1393 M10, pVL1393 Mi l, pVL1393 M12, FLAG vectors such as pPolh-FLAGl or pPolh-MAT 2, or MAT vectors such as pPolh-MATl, or pPolh-MAT2.
  • FLAG vectors such as pPolh-FLAGl or pPolh-MAT 2
  • MAT vectors such as pPolh-MATl, or pPolh-MAT
  • a FGF mutant described herein is used for the treatment of a metabolic disease or condition, such as an FGFl -related disease or condition or a FGF2-related disease or condition. In some instances, a FGF mutant described herein is used for the treatment of an FGF1- related disease or condition.
  • administration of a FGF mutant described herein reduces blood glucose, improves insulin sensitivity, improves glucose tolerance, reduces systemic chronic inflammation, ameliorate hepatic steatosis, reduces food intake, or combinations thereof.
  • administration of a FGF mutant described herein reduces blood glucose.
  • administration of a FGF mutant described herein improves insulin sensitivity.
  • administration of a FGF mutant described herein improves glucose tolerance.
  • administration of a FGF mutant described herein reduces systemic chronic inflammation.
  • administration of a FGF mutant described herein ameliorate hepatic steatosis.
  • administration of a FGF mutant described herein reduces food intake.
  • Type 1 diabetes also called insulin-dependent diabetes, comprises about 5% to 10% of all diabetes cases.
  • Type 1 diabetes is an autoimmune disease where T cells attack and destroy insulin- producing beta cells in the pancreas.
  • Type 1 diabetes is caused by genetic and environmental factors.
  • double diabetes is used to describe patients diagnosed with both type 1 and 2 diabetes.
  • Type 4 diabetes is a recently discovered type of diabetes affecting about 20% of diabetic patients age 65 and over. In some embodiments, type 4 diabetes is characterized by age-associated insulin resistance.
  • Diabetes insipidus is a rare disorder affecting 3 in about 100,000 people each year worldwide.
  • diabetes insipidus is further classified into neurogenic DI, nephrogenic DI, dipsogenic DI, and gestational DI.
  • neurogenic DI also known as central DI, is characterized by damage to the pituitary gland, which results in disruption of antidiuretic hormone (ADH) storage and release.
  • ADH antidiuretic hormone
  • nephrogenic DI is characterized by the inability of the kidneys to respond to ADH.
  • dipsogenic DI is characterized by a defect or damage in the thirst mechanism governed by the hypothalamus, which results in suppression of ADH and an increase in urine output.
  • gestational DI is characterized by degradation of vasopressin in the mother by placental vasopressinase.
  • an FGFl mutant described herein is used for the treatment of type 1 diabetes, type 2 diabetes, type 3 diabetes, type 4 diabetes, double diabetes, latent autoimmune diabetes (LAD), gestational diabetes, neonatal diabetes mellitus (NDM), maturity onset diabetes of the young (MODY), Wolfram syndrome, Alstrom syndrome, prediabetes, or diabetes insipidus.
  • LAD latent autoimmune diabetes
  • NDM neonatal diabetes mellitus
  • MODY maturity onset diabetes of the young
  • an FGFl mutant described herein is used for the treatment of type 1 diabetes.
  • an FGFl mutant described herein is used for the treatment of type 2 diabetes.
  • an FGFl mutant described herein is used for the treatment of type 3 diabetes.
  • an FGFl mutant described herein is used for the treatment of type 4 diabetes. In some cases, an FGFl mutant described herein is used for the treatment of double diabetes. In some cases, an FGFl mutant described herein is used for the treatment of latent autoimmune diabetes (LAD) (e.g., LADA, LADY, or LADC). In some cases, an FGFl mutant described herein is used for the treatment of gestational diabetes. In some cases, an FGFl mutant described herein is used for the treatment of neonatal diabetes mellitus (NDM) (e.g., PNDM or TNDM).
  • NDM neonatal diabetes mellitus
  • a FGF mutant described herein is used for the treatment of NAFLD.
  • the FGF mutant is an FGF 1 mutant.
  • an FGF 1 mutant described herein is used for the treatment of NAFLD.
  • a FGF mutant described herein is used for the treatment of lipodystrophy.
  • the FGF mutant is an FGF1 mutant.
  • an FGF1 mutant described herein is used for the treatment of lipodystrophy.
  • a metabolic disease or condition is a cardiovascular disease.
  • a FGF mutant e.g., FGF1 mutant
  • the additional therapeutic agent is a therapeutic agent for the treatment of a metabolic disease selected from diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic steatohepatitis (NASH), nonalcoholic fatty liver disease (NAFLD), dyslipidemia, a cardiovascular disease, Hurler syndrome, Huntington's disease, Alzheimer's disease, Parkinson's disease, Crohn's disease, Fabry disease, and galactosemia.
  • a metabolic disease selected from diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic steatohepatitis (NASH), nonalcoholic fatty liver disease (NAFLD), dyslipidemia, a cardiovascular disease, Hurler syndrome, Huntington's disease, Alzheimer's disease, Parkinson's disease, Crohn's disease, Fabry disease, and galactosemia.
  • a FGF mutant (e.g., FGF1 mutant) described herein is used in combination with a therapeutic agent for the treatment of a metabolic disease selected from diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic alcoholic fibrosis, diabetes, diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic fibrosis, diabetes, diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic ovary syndrome (PCOS), diabetes, diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic a metabolic disease, and others.
  • the additional therapeutic agent is a therapeutic agent for the treatment of Crohn's disease and includes: 5 -aminosalicylates such as sulfasalazine or mesalamine; corticosteroids such as prednisone, methylprednisolone, hydrocortisone, or budesonide; immune system suppressors such as azathioprine, mercaptopurine, infliximab, adalimumab, certolizumab pegol, methotrexate, cyclosporine, tracrolimus, natalizumab, vedolizumab, or ustekinumab;
  • 5 -aminosalicylates such as sulfasalazine or mesalamine
  • corticosteroids such as prednisone, methylprednisolone, hydrocortisone, or budesonide
  • immune system suppressors such as azathioprine, mercaptopur
  • the additional therapeutic agent is a therapeutic agent for the treatment of Fabry disease and includes: agalsidase beta or alpha-galactosidase.
  • the additional therapeutic agent is a glucagon-like peptide- 1 receptor
  • a FGF mutant described herein is modified by myristoylation, palmitoylation, isoprenylation (or prenylation) (e.g., farnesylation or geranylgeranylation), glypiation, acylation (e.g., O-acylation, N-acylation, S-acylation), alkylation (e.g., additional of alkyl groups such as methyl or ethyl groups), amidation, glycosylation, hydroxylation, iodination, nucleotide addition, oxidation, phosphorylation, succinylation, sulfation, glycation, carbamylation, glutamylation, or deamidation.
  • myristoylation, palmitoylation, isoprenylation (or prenylation) e.g., farnesylation or geranylgeranylation
  • acylation e.g., O-acylation, N-acylation, S-acylation
  • alkylation
  • a conjugating moiety comprises a polymer.
  • a polymer is a natural or synthetic polymer, consisting of, e.g., long chains of branched or unbranched monomers, and/or cross-linked network of monomers in two or three dimensions.
  • a polymer comprises a water soluble polymer or a water insoluble polymer.
  • a FGF polypeptide conjugate comprises a FGF-polymer conjugate.
  • a polymer conjugate alters one or more properties (e.g., biological properties and/or physicochemical properties) of a FGF polypeptide.
  • polyamination modification of a FGF polypeptide described herein by a polyamine is referred to as polyamination.
  • polyamination is an enzyme -mediated conjugation, in which transglutaminase (Tgase) catalyzes the transfer of a primary amino group to an acyl group.
  • Tgase transglutaminase
  • polyamination is as described in Zhang, et al, "Synthesis and applications of polyamine amino acid residues: improving the bioactivity of an analgesic neuropeptide, Neurotensin," J Med Chem 52(6): 1514-1517 (2009).
  • a FGF polypeptide described herein is modified by polyamination.
  • a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974 is modified by polyamination.
  • a polymer comprises a lipid moiety.
  • Lipid is a broad family of small molecules capable of forming structures such as vesicles, multilamellar/unilamellar liposomes, or membranes in an aqueous environment.
  • a lipid comprises a fatty acid.
  • a lipid comprises a short chain fatty acid (less than C6), a medium chain fatty acid (C6-C14), or a long chain fatty acid (C15 or longer).
  • a fatty acid comprises a saturated fatty acid or an unsaturated fatty acid.
  • a lipid comprises an unsaturated fatty acid.
  • unsaturated fatty acid comprises monounsaturated fatty acid such as palmitoleic acid, vaccenic acid, oleic acid, eicosenoic acid, erucic acid, gadoleic acid, myristoleic acid, or nervonic acid; and polyunsaturated fatty acid such as hexadecatrienoic acid, alpha-linolenic acid, stearidonic acid, eicosatrienoic acid, eicosatetraenoic acid, eicosapentaenoic acid, heneicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, tetracosapentaenoic acid, tetracosahexaenoic acid, linoleic acid, gamma-linolenic acid,
  • a FGF-polymer conjugate comprises a FGF-lipid conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-fatty acid conjugate.
  • a FGF polypeptide described herein is conjugated to a functional fragment of albumin.
  • a functional fragment of albumin comprises an albumin with a N-terminal truncation, a C-terminal truncation or a truncation of both termini.
  • a functional fragment of albumin comprises domain I, domain II, domain III, or a combination thereof.
  • a functional fragment of albumin comprises a polypeptide of at least 70%, 80%, 90%, or 95% sequence identity to the full-length wild type albumin (e.g., HSA).
  • a FGF polypeptide described herein is conjugated to a functional fragment of HSA.
  • a conjugating moiety comprises a conjugating peptide.
  • a conjugating peptide modulates one or more biological properties and/or physicochemical properties of a target molecule of interest (e.g., a FGF polypeptide).
  • a conjugating peptide enhances pharmacokinetics, pharmacodynamics, metabolic stability, biodistribution, water solubility, lipophilicity, or tissue targeting capabilities of a target molecule of interest (e.g., a FGF polypeptide).
  • a XTENTM peptide is selected based on properties such as expression, genetic stability, solubility, aggregation resistance, enhanced half- life, increased potency, and/or increased in vitro activity in combination with a polypeptide of interest.
  • a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974 is modified by XTENylation.
  • modification of a FGF polypeptide described herein by an elastin- like polypeptide (ELP) is referred to as ELPylation.
  • a FGF polypeptide described herein is conjugated to an antibody or its binding fragments thereof.
  • a FGF polypeptide described herein is conjugated to an antibody or its binding fragments thereof selected from a humanized antibody or binding fragment thereof, murine antibody or binding fragment thereof, chimeric antibody or binding fragment thereof, monoclonal antibody or binding fragment thereof, monovalent Fab', divalent Fab 2 , F(ab)'3 fragments, single-chain variable fragment (scFv), bis-scFv, (scFv) 2 , diabody, minibody, nanobody, triabody, tetrabody, humabody, disulfide stabilized Fv protein (dsFv), single-domain antibody (sdAb), Ig NAR, camelid antibody or binding fragment thereof, bispecific antibody or biding fragment thereof, or a chemically modified derivative thereof.
  • a FGF polypeptide selected from SEQ ID NOs: 3-25917 and 25922-25974 is conjugated to an antibody or its binding fragments thereof selected from a humanized antibody or binding fragment thereof, murine antibody or binding fragment thereof, chimeric antibody or binding fragment thereof, monoclonal antibody or binding fragment thereof, monovalent Fab', divalent Fab 2 , F(ab)'3 fragments, single-chain variable fragment (scFv), bis-scFv, (scFv) 2 , diabody, minibody, nanobody, triabody, tetrabody, humabody, disulfide stabilized Fv protein (dsFv), single-domain antibody (sdAb), Ig NAR, camelid antibody or binding fragment thereof, bispecific antibody or biding fragment thereof, or a chemically modified derivative thereof.
  • the conjugation is a covalent conjugation.
  • the conjugation is genetic linkage of an antibody or its binding fragments thereof to a FGF polypeptide.
  • Affimer is derived from cystatins, a family of cysteine protease inhibitors.
  • the Affimer structure comprises two peptide loops and an N-terminal sequence which in some cases are randomized to generate a target site of interest.
  • Alphabodies or cell-penetrating alphabodies or CPAB
  • Alphabodies are small proteins of about 10 kDa in size.
  • Alphabodies comprise coiled-coil structures and the sequences are modified, e.g., by phase display, to generate target site of interest.
  • Avimers comprise two or more peptide sequences of about 30 to about 35 amino acid residues in length.
  • each peptide sequence interacts with an epitope of a target protein of interest, and the combination of peptides to different epitopes of a target protein of interest increases affinity to the target protein.
  • a nanoparticle includes a core or a core and a shell, as in a core-shell nanoparticle.
  • a nanoparticle has at least one dimension of less than about 500nm, 400nm, 300nm, 200nm, or lOOnm.
  • a FGF mutant is encapsulated within a hydrogel or a scaffold.
  • the hydrogels or scaffolds are comprised of synthetic polymers such as, but not limited to, poly(a-hydroxy acids), poly(orthoesters), poly(anhydrides), poly(amino acids), dextrin, poly(glycoside) (PGA), poly(L-lactide) (PLA) and their copolymers poly(D,L-lactic-co-glycolic) acid (PLGA); naturally occurring polymers such as, but not limited to, silk, keratin, collagen, denatured collagen, atelocollagen, gelatin, fibrinogen, fibrin, laminin, elastin, chitosan, hyaluronic acid, starch, carrageenan, cellulose, alginate, perlecan (PLC), heparin or heparin sulfate, chondroitin sulfate, fibronectin, and
  • the pharmaceutical formulations include, but are not limited to, aqueous liquid dispersions, self-emulsifying dispersions, solid solutions, liposomal dispersions, aerosols, solid dosage forms, powders, immediate release formulations, controlled release formulations, fast melt formulations, tablets, capsules, pills, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations (e.g., nanoparticle formulations), and mixed immediate and controlled release formulations.
  • aqueous liquid dispersions self-emulsifying dispersions, solid solutions, liposomal dispersions, aerosols, solid dosage forms, powders, immediate release formulations, controlled release formulations, fast melt formulations, tablets, capsules, pills, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations (e.g., nanoparticle formulations), and mixed immediate and controlled release formulations.
  • Pharmaceutically compatible carrier materials include, but are not limited to, acacia, gelatin, colloidal silicon dioxide, calcium glycerophosphate, calcium lactate, maltodextrin, glycerine, magnesium silicate, polyvinylpyrrollidone (PVP), cholesterol, cholesterol esters, sodium caseinate, soy lecithin, taurocholic acid, phosphotidylcholine, sodium chloride, tricalcium phosphate, dipotassium phosphate, cellulose and cellulose conjugates, sugars sodium stearoyl lactylate, carrageenan, monoglyceride, diglyceride, pregelatinized starch, and the like. See, e.g., Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack
  • the pharmaceutical formulations include, but are not limited to, sugars like trehalose, sucrose, mannitol, maltose, glucose, or salts like potassium phosphate, sodium citrate, ammonium sulfate and/or other agents such as heparin to increase the solubility and in vivo stability of polypeptides.
  • Such compounds can include e.g., lactose, starch, mannitol, sorbitol, dextrose, microcrystalline cellulose such as Avicel ® , dibasic calcium phosphate, dicalcium phosphate dihydrate, tricalcium phosphate, calcium phosphate, anhydrous lactose, spray-dried lactose, pregelatinized starch, compressible sugar, such as Di-Pac ® (Amstar), mannitol,
  • hydroxypropylmethylcellulose hydroxypropylmethylcellulose acetate stearate, sucrose-based diluents, confectioner's sugar, monobasic calcium sulfate monohydrate, calcium sulfate dihydrate, calcium lactate trihydrate, dextrates, hydrolyzed cereal solids, amylose, powdered cellulose, calcium carbonate, glycine, kaolin, mannitol, sodium chloride, inositol, bentonite, and the like.
  • the pharmaceutical formulations include disintegration agents or disintegrants to facilitate the breakup or disintegration of a substance.
  • disintegrate include both the dissolution and dispersion of the dosage form when contacted with gastrointestinal fluid.
  • Lubricants and glidants are also optionally included in the pharmaceutical formulations described herein for preventing, reducing or inhibiting adhesion or friction of materials.
  • Exemplary lubricants include, e.g., stearic acid, calcium hydroxide, talc, sodium stearyl fumerate, a hydrocarbon such as mineral oil, or hydrogenated vegetable oil such as hydrogenated soybean oil (Sterotex ® ), higher fatty acids and their alkali-metal and alkaline earth metal salts, such as aluminum, calcium, magnesium, zinc, stearic acid, sodium stearates, glycerol, talc, waxes, Stearowet ® , boric acid, sodium benzoate, sodium acetate, sodium chloride, leucine, a polyethylene glycol (e.g., PEG-4000) or a methoxypolyethylene glycol such as CarbowaxTM, sodium oleate, sodium benzoate, glyceryl behenate, polyethylene
  • Cab-O-Sil ® a starch such as corn starch, silicone oil, a surfactant, and the like.
  • Plasticizers include compounds used to soften the microencapsulation material or film coatings to make them less brittle. Suitable plasticizers include, e.g., polyethylene glycols such as
  • Solubilizers include compounds such as triacetin, triethylcitrate, ethyl oleate, ethyl caprylate, sodium lauryl sulfate, sodium doccusate, vitamin E TPGS, dimethylacetamide, N- methylpyrrolidone, N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropylmethyl cellulose, hydroxypropyl cyclodextrins, ethanol, n-butanol, isopropyl alcohol, cholesterol, bile salts, polyethylene glycol 200-600, glycofurol, transcutol, propylene glycol, and dimethyl isosorbide and the like.
  • Suspending agents include compounds such as polyvinylpyrrolidone, e.g.,
  • kits and articles of manufacture for use with one or more methods and compositions described herein.
  • Such kits include a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method described herein.
  • Suitable containers include, for example, bottles, vials, syringes, and test tubes.
  • the containers are formed from a variety of materials such as glass or plastic.
  • a label is on or associated with the container.
  • a label is on a container when letters, numbers or other characters forming the label are attached, molded or etched into the container itself, a label is associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert.
  • the pharmaceutical compositions are presented in a pack or dispenser device which contains one or more unit dosage forms containing a compound provided herein.
  • the pack for example, contains metal or plastic foil, such as a blister pack.
  • the terms "individual(s)", “subject(s)” and “patient(s)” mean any mammal.
  • the mammal is a human.
  • the mammal is a non-human. None of the terms require or are limited to situations characterized by the supervision (e.g. constant or intermittent) of a health care worker (e.g. a doctor, a registered nurse, a nurse practitioner, a physician's assistant, an orderly or a hospice worker).
  • a health care worker e.g. a doctor, a registered nurse, a nurse practitioner, a physician's assistant, an orderly or a hospice worker.

Abstract

Disclosed herein are FGF compositions and methods of treating a metabolic disease or condition. Also described herein are FGF1 mutants, pharmaceutical formulations, and their use in the treatment of a metabolic disease or condition.

Description

FGF MUTANTS AND USES THEREOF
CROSS-REFERENCE
[0001] This application claims the benefit of US Provisional Application No. 62/365,296, filed on July 21, 2016, which is incorporated herein by reference in its entirety.
SEQUENCE LISTING
[0002] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on July 20, 2017, is named 48773-809_601_SL.txt and is 32,716,578 bytes in size.
BACKGROUND OF THE DISCLOSURE
[0003] Metabolic diseases or conditions such as diabetes and obesity are growing health problems worldwide. In some instances, it is estimated that more than 250 million adults worldwide have diabetes and the figure is projected to increase to 380 million by 2025. The burden of such metabolic diseases or conditions such as diabetes and obesity, in terms of the prevalence and number of individuals affected, as well as in terms of the complexity in disease etiology, present challenges in treatments to slow disease progression and achieve metabolic control.
SUMMARY OF THE DISCLOSURE
[0004] Described herein, in certain embodiments, include FGF mutants and their use in the treatment of one or more metabolic diseases or conditions. In some instances, the FGF mutant is an FGF1 mutant. In some cases, an FGF1 mutant is used for the treatment of one or more metabolic diseases or conditions.
[0005] Disclosed herein, in certain embodiments, is an isolated and purified FGF polypeptide comprising at least 80% sequence identity to SEQ ID NOs: 3-25917 and 25922-25974. In some embodiments, the FGF polypeptide comprises at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NOs: 3-25917 and 25922-25974. In some embodiments, the FGF polypeptide consists of SEQ ID NOs: 3-25917 and 25922-25974. In some embodiments, the FGF polypeptide comprises at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NOs: 130, 131, 132, 133, 134, 135, 139, 141, 142, 143, 144, 146, 147, 148, 149, 150, 151, 152, 153, 156, 157, 158, 159, 163, 165, 168, 170, 171, 174, 177, 284, 286, 290, 401, 484, 488, or 518. In some embodiments, the FGF polypeptide further comprises a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2. In some embodiments, the FGF polypeptide further comprises a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L89, T96, K128, or a combination thereof, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2. In some embodiments, the FGF polypeptide further comprises a modification at amino acid residue L84, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2. In some embodiments, the modification comprises a mutation to a non-polar residue or a charged residue. In some embodiments, the modification comprises a mutation to alanine. In some embodiments, the isolated and purified FGF polypeptide comprises a modification of T30A, T34A, A48G, E53A, K57A, K57E, Q63A, T78A, P79A, L84A, L89A, T96A, K128E, or a combination thereof. In some embodiments, the isolated and purified FGF polypeptide has enhanced stability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild- type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide is an FGF1 mutant.
[0006] Disclosed herein, in some embodiments, is an isolated and purified FGF polypeptide comprising a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2. In some embodiments, the FGF polypeptide comprises a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L89, T96, K128, or a combination thereof, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2. In some embodiments, the FGF polypeptide comprises a modification at amino acid residue L84, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2. In some embodiments, the isolated and purified FGF polypeptide further comprises a modification at amino acid residue H41, 142, or Q43, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2. In some embodiments, the isolated and purified FGF polypeptide further comprises a modification at amino acid residue K10, Y15, S17, N18, H21, L26, P27, D28, R35, D36, R37, S38, D39, Q40, Q43, S47, Y55, E60, T61, Y64, D68, E81, E87, R88, E90, E91, N92, Y94, N95, K100, K101, H102, K105, Kl 13, S I 16, CI 17, Kl 18, P121, R122, H124, Q127, or L133, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2. In some embodiments, the modification comprises a mutation to a non-polar residue or a charged residue. In some embodiments, the modification comprises a mutation to alanine. In some embodiments, the isolated and purified FGF polypeptide comprises a modification of T30A, T34A, A48G, E53A, K57A, K57E, Q63A, T78A, P79A, L84A, L89A, T96A, K128E, or a combination thereof. In some embodiments, the isolated and purified FGF polypeptide further comprises at least one additional modification. In some embodiments, the at least one additional modification comprises an insertion, a deletion, or a mutation. In some embodiments, the mutation comprises a mutation to a polar residue, a non-polar residue, or a charged residue. In some embodiments, the isolated and purified FGF polypeptide further comprises a truncation. In some embodiments, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some embodiments, the isolated and purified FGF polypeptide has enhanced stability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild- type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide is an FGF1 mutant.
[0007] Disclosed herein, in certain embodiments, is an isolated and purified FGF polypeptide comprising: one or more modified amino acids selected from Group 1 amino acid residues: P27, R35, D36, R37, Y55, K57, T61, E81, E87, R88, N92, T96, K101, H102, and L133; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; wherein the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some embodiments, the one or more modified amino acids from Group 1 is selected from Group 1A amino acid residues: R35, D36, and R37. In some embodiments, the one or more modified amino acids from Group 1 is selected from Group IB amino acid residues: E87, R88, N92, T96, and L133. In some embodiments, the one or more modified amino acids from Group 1 is selected from Group 1C amino acid residues: P27, Y55, K57, T61, E81, K101, and H102. In some embodiments, the one or more modified amino acids from Group 1 is R35, N92, or T96. In some embodiments, the one or more modified amino acids from Group 1 is R35, and the one or more modified amino acids from Group 2 is selected from Q43, Q77, N80, and E82. In some embodiments, the one or more modified amino acids from Group 1 is N92, and the one or more modified amino acids from Group 2 is Q43. In some embodiments, the one or more modified amino acids from Group 1 is T96, and the one or more modified amino acids from Group 2 is Q43. In some embodiments, the one or more modified amino acids comprise a mutation to a non-polar residue or a charged residue. In some embodiments, the one or more modified amino acids comprise a mutation to alanine. In some embodiments, the FGF polypeptide further comprises a truncation. In some embodiments, the truncation is an N-terminal truncation. In some embodiments, the N-terminal truncation is a deletion of amino acid residues 1-9. In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide.
[0008] Disclosed herein, in certain embodiments, is an isolated and purified FGF polypeptide comprising two or more modifications, the polypeptide comprising: a first modification at an amino acid residue selected from: Y15, R35, R37, K57, R88, N92, T96, R122, and L133; and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S 17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, SI 16, Rl 19, H124, Y125, P136, V137, S138, S139, and D140; wherein the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some embodiments, the second modification is at an amino acid residue selected from: N7, S38, Q43, Q45, A48, S58, T78, P79, N80, E81, E82, E91, H93, 198, K101, P136, and V137. In some embodiments, the first modification is at R35, and the second modification is at an amino acid residue selected from the group consisting of: Fl, N2, P5, G6, N7, K9, K10, S 17, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, and D140. In some embodiments, the first modification is at R35, and the second modification is at an amino acid residue selected from the group consisting of: A48, E91, N92, H93, 198, K101, and V137. In some embodiments, the first modification is at R35, and the second modification is at an amino acid residue selected from the group consisting of: Fl, N2, P5, K10, R37, Q45, S47, E49, S50, V51, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E104, SI 16, Y125, P136, and S 139. In some embodiments, the first modification is at R35, and the second modification is at an amino acid residue selected from the group consisting of: G6, K9, and S17. In some embodiments, the first modification is at N92, and the second modification is at an amino acid residue selected from the group consisting of R37 and Q43. In some embodiments, the first modification is at R37, and the second modification is at an amino acid residue selected from the group consisting of: Q43, E53, and T96. In some embodiments, the first modification is at R37, and the second modification is at an amino acid residue selected from the group consisting of N92 and T96. In some embodiments, the first modification is at R37, and the second modification is at an amino acid residue selected from the group consisting of: Q43 and E53. In some embodiments, the first modification is at R88, and the second modification is at an amino acid residue selected from the group consisting of E91 and T96. In some embodiments, the first modification is at K57, and the second modification is at an amino acid residue selected from the group consisting of: K10, A48, E90, and E91. In some embodiments, the first modification is at K57, and the second modification is at an amino acid residue selected from the group consisting of: K10, A48, and E90. In some embodiments, the first modification is at T96, and the second modification is at Q43. In some embodiments, the first modification is at Y 15, and the second modification is at Q43. In some embodiments, the first modification is at R122, and the second modification is at an amino acid residue selected from the group consisting of: R35, R37, and K57. In some embodiments, the FGF polypeptide further comprises a truncation. In some embodiments, the truncation is an N-terminal truncation. In some embodiments, the N-terminal truncation is a deletion of amino acid residues 1-9. In some embodiments, the modification comprises a mutation to a non- polar residue or a charged residue. In some embodiments, the modification comprises a mutation to alanine. In some embodiments, Y15, R35, R37, K57, R88, N92, T96, and R122 are independently mutated to an amino acid selected from the group consisting of: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some embodiments, Y15 is mutated to phenylalanine (F). In some embodiments, R35 is mutated to an amino acid selected from the group consisting of: alanine (A) and glutamic acid (E). In some embodiments, R37 is mutated to an amino acid selected from the group consisting of: alanine (A) and glutamic acid (E). In some embodiments, K57 is mutated to an amino acid selected from the group consisting of: alanine (A), asparagine (N), glutamic acid (E), and histidine (H). In some embodiments, R88 is mutated to leucine (L). In some embodiments, N92 is mutated to an amino acid selected from the group consisting of: alanine (A) and threonine (T). In some embodiments, T96 is mutated to alanine (A). In some embodiments, the FGF polypeptide is an FGFl polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide.
[0009] Disclosed herein, in certain embodiments, is an isolated and purified FGF polypeptide comprising a modification at amino acid residue A48, E53, K57, S58, L72, S76, Q77, T78, P79, T96, or 198, wherein the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some embodiments, the modification is at K57. In some embodiments, the modification is at Q77. In some embodiments, the modification is at T96. In some embodiments, the FGF polypeptide further comprises a truncation. In some embodiments, the truncation is an N-terminal truncation. In some embodiments, the N-terminal truncation is a deletion of amino acid residues 1-9. In some
embodiments, the modification comprises a mutation to a non-polar residue or a charged residue. In some embodiments, the modification comprises a mutation to alanine. In some embodiments, the FGF polypeptide is an FGFl polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide.
[0010] Disclosed herein, in certain embodiments, is an isolated and purified FGF polypeptide comprising at least 80% sequence identity to a sequence selected from SEQ ID NOs: 3-25917 and 25922-25974. In some embodiments, the FGF polypeptide comprises at least 85%, 90%, 95%, 96%), 97%), 98%o, 99%o, or more sequence identity to a sequence selected from SEQ ID NOs: 3- 25917 and 25922-25974. In some embodiments, the FGF polypeptide consists of a sequence selected from SEQ ID NOs: 3-25917 and 25922-25974. In some embodiments, the FGF polypeptide is an FGFl polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide.
[0011] Disclosed herein, in certain embodiments, is an isolated and purified FGF polypeptide having an FGFR interaction activity that is equivalent to the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2; and a mitogenicity that is at least 1-fold lower than the mitogenicity of the FGFl polypeptide of SEQ ID NO: 2. In some embodiments, the FGFR interaction activity of the isolated and purified FGF polypeptide is about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% to the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some embodiments, the mitogenicity is at least 2-fold lower, 3 -fold lower, 4-fold lower, 5 -fold lower, 6-fold lower, 7-fold lower, 8-fold lower, 9-fold lower, or 10-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2. In some embodiments, the isolated and purified FGF polypeptide is an isolated and purified FGF polypeptide described supra. In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide. In some embodiments, the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some embodiments, the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide.
[0012] Disclosed herein, in certain embodiments, is a pharmaceutical composition comprising an isolated and purified FGF polypeptide disclosed above; and an excipient. In some embodiments, the composition further comprises at least one additional FGF polypeptide, wherein the at least one additional FGF polypeptide is identical to the FGF polypeptide. In some embodiments, the composition further comprises at least one additional FGF polypeptide, wherein the at least one additional FGF polypeptide is not identical to the FGF polypeptide. In some embodiments, the at least one additional FGF polypeptide comprises at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NOs: 1-25917 and 25922-25974. In some embodiments, the at least one additional FGF polypeptide comprises at least one modification, a truncation, or a combination thereof. In some embodiments, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some embodiments, the pharmaceutical composition is formulated for oral or parenteral administration.
[0013] A method of treating a metabolic disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an isolated and purified FGF polypeptide described above. In some embodiments, the metabolic disease or condition comprises FGF 1 -related disease or condition. In some embodiments, the metabolic disease or condition comprises diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), dyslipidemia, a cardiovascular disease, Hurler syndrome, Huntington's disease, Alzheimer's disease, Parkinson's disease, Crohn's disease, Fabry disease, or galactosemia. In some embodiments, the metabolic disease or condition comprises impaired insulin sensitivity or glucose intolerance. In some embodiments, the subject has diabetes. In some embodiments, the diabetes comprises type 1 diabetes, type 2 diabetes, type 3 diabetes, type 4 diabetes, double diabetes, latent autoimmune diabetes (LAD), gestational diabetes, neonatal diabetes mellitus (NDM), maturity onset diabetes of the young
(MODY), Wolfram syndrome, Alstrom syndrome, prediabetes, or diabetes insipidus. In some embodiments, the subject has type 1 diabetes. In some embodiments, the subject has type 2 diabetes. In some embodiments, the subject has type 4 diabetes. In some embodiments, the subject has prediabetes. In some embodiments, the subject has polycystic ovary syndrome. In some embodiments, the subject has metabolic syndrome. In some embodiments, the subject has obesity. In some embodiments, the subject has non-alcoholic fatty liver disease. In some embodiments, the subject has insulin resistance. In some embodiments, the subject has lipodystrophy. In some embodiments, the subject has a cardiovascular disease. In some embodiments, the cardiovascular disease comprises hypertension, coronary artery diseases (CAD), stroke, rheumatic heart disease, hypertensive heart disease, cardiomyopathy, heart arrhythmia, congenital heart disease, carditis, aortic aneurysms, renal artery stenosis, peripheral artery disease, venous thrombosis, heart valve disease, deep vein thrombosis, cerebrovascular disease, cardiac dysrhythmia, chronic obstructive pulmonary disease, or inflammatory heart disease. In some embodiments, the subject has hypertension. In some
embodiments, the method further comprises administration of an additional therapeutic agent. In some embodiments, the additional therapeutic agent comprises thiazolidinedione. In some embodiments, thiazolidinedione comprises pioglitazone, rosiglitazone, lobeglitazone, and troglitazone. In some embodiments, the FGF polypeptide and the additional therapeutic agent are administered
simultaneously. In some embodiments, the FGF polypeptide and the additional therapeutic agent are administered sequentially. In some embodiments, the FGF polypeptide is administered prior to the administration of the additional therapeutic agent. In some embodiments, the FGF polypeptide is administered after the administration of the additional therapeutic agent. In some embodiments, the FGF polypeptide and the additional therapeutic agent are formulated in a unified dosage form. In some embodiments, the FGF polypeptide and the additional therapeutic agent are formulated in separate dosage forms.
[0014] Disclosed herein, in certain embodiments, is a polynucleic acid molecule encoding an FGF polypeptide described above.
[0015] Disclosed herein, in certain embodiments, is a kit comprising an isolated and purified FGF polypeptide described above.
[0016] Disclosed herein, in certain embodiments, is a kit comprising a polynucleic acid molecule encoding an FGF polypeptide described above.
BRIEF DESCRIPTION OF THE DRAWINGS [0017] Various aspects of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings of which:
[0018] Fig. lA-Fig. IB illustrate the location of the FGF1 Group 1 residues with respect to a FGFl-FGFRlc complex (PDB: 30JV.pdb) in a first orientation. Fig. 1A shows the FGFl-FGFRlc complex and Fig. IB shows a close-up of the positions of the Group 1A, Group IB, and Group 1C residues on FGF1.
[0019] Fig. 2A-Fig. 2B illustrate the location of the FGF1 Group 1 residues with respect to a FGFl-FGFRlc complex (PDB: 30JV.pdb) in a second orientation. Fig. 2A shows the FGFl-FGFRlc complex and Fig. 2B shows a close-up of the positions of the Group 1A, Group IB, and Group 1C residues on FGF1.
[0020] Fig. 3A-Fig. 3B illustrate the location of the FGF1 Group 1 and Group 2 residues with respect to a FGFl-FGFRlc complex (PDB: 30JV.pdb) in a first orientation. Fig. 3A shows the FGFl-FGFRlc complex and Fig. 3B shows a close-up of the positions of the Group 1 and Group 2 residues on FGF1.
[0021] Fig. 4A-Fig. 4B illustrate the location of the FGF1 Group 1 and Group 2 residues with respect to a FGFl-FGFRlc complex (PDB: 30JV.pdb) in a second orientation. Fig. 4A shows the FGFl-FGFRlc complex and Fig. 4B shows a close-up of the positions of the Group 1 and Group 2 residues on FGF1.
[0022] Fig. 5A - Fig. 5D illustrate the location of exemplary FGF1 site of mutation with respect to a FGFl-FGFRlc complex (PDB: 30JV.pdb). Fig. 5A and Fig. 5C show the FGFl-FGFRlc complex. Fig. 5B and Fig. 5D show close-ups of the positions of the exemplary FGF1 residues (Fig. 5B) and their position relative to Group 1 residues (Fig. 5D).
[0023] Fig. 6 illustrates the mitogenic activity of exemplary FGF1 polypeptides in zona glomerulosa.
[0024] Fig. 7 illustrates the mitogenic activity of exemplary FGF1 polypeptides in zona fasciculata/reticularis.
[0025] Fig. 8 illustrates the mitogenic activity of exemplary FGF1 polypeptides in medulla.
[0026] Fig. 9 illustrates the mitogenic activity of exemplary FGF1 polypeptides in hepatocytes.
[0027] Fig. 1 OA-Fig. 10B illustrate glucose level and insulin sensitivity of mice to exemplary FGF1 polypeptides. Fig. 10A illustrates blood glucose level at 0 hour, 4 hours, and 24 hours of exemplary FGF1 polypeptides: MP516, MP2744, MP2779, MP2786, MP2847, and MP2850. Fig. 10B shows the HOMA-IR measurement at Week 0, Week 2, and Week 3.
[0028] Fig. 1 lA-Fig. 1 IB illustrate glucose level and insulin sensitivity of mice to exemplary FGF1 polypeptides. Fig. 11A illustrates blood glucose level at 0 hour, 4 hours, and 24 hours of exemplary FGF1 polypeptides: MP607, MP222, MP242, MP518, and MP220. Fig. 11B shows the HOMA-IR measurement at Week 0, Week 2, and Week 3.
[0029] Fig. 12A-Fig. 12B illustrate glucose level and insulin sensitivity of mice to exemplary FGF1 polypeptides. Fig. 12A illustrates blood glucose level at 0 hour, 4 hours, and 24 hours of exemplary FGF1 polypeptides: MP200, MP266, and MP1348. Fig. 12B shows the HOMA-IR measurement at Week 0, Week 2, and Week 3.
[0030] Fig. 13A-Fig. 13C illustrate in-vivo mitogenicity and in-vivo efficacy of exemplary FGF1 polypeptides. Fig. 13A shows the in-vivo mitogenicity of exemplary FGF1 polypeptides. Fig. 13B and Fig. 13C show the in-vivo efficacy of exemplary FGF1 polypeptides.
DETAILED DESCRIPTION OF THE DISCLOSURE
[0031] Fibroblast growth factors (FGF) are a family of growth factors that are involved in endocrine signaling pathways, wound healing, angiogenesis, and embryonic development. There are 22 structurally related members in the FGF family, which are further classified as intracrine, paracrine, and endocrine FGFs, based on their action mechanism. Paracrine and endocrine FGFs are extracellular signaling molecules that interact with fibroblast growth factor receptors (FGFRs).
Paracrine FGF members include FGF1, FGF2, FGF3, FGF4, FGF5, FGF6, FGF7, FGF 8, FGF9, FGF 10, FGF 16, FGF 17, FGF 18, FGF20, and FGF22. Endocrine FGF members include
FGF19/FGF15 (FGF 15 is the mouse homolog of human FGF 19), FGF21, and FGF23. Intracrine FGFs are FGFR-independent intracellular molecules that regulate the function of voltage gated sodium channels and the members include FGF11, FGF12, FGF13, and FGF14.
[0032] In some cases, FGF1 and FGF2 are further grouped into the FGF1 subfamily. Fibroblast growth factor 1 (FGF1) is the founding member of the FGF1 subfamily and the FGF family and is involved in cell growth, proliferation, migration, differentiation, and survival. Also known as acidic fibroblast growth factor or acidic heparin binding growth factor, FGF 1 is an autocrine/paracrine regulator. Unlike the other members of the paracrine FGF subgroup, FGF1 interacts with all four FGFRs, fibroblast growth factor receptor 1 (FGFR1), fibroblast growth factor receptor 2 (FGFR2), fibroblast growth factor receptor 3 (FGFR3), and fibroblast growth factor receptor 4 (FGFR4), and in particular, isoforms FGFRlb (FGFRlIIIb), FGFRlc (FGFRlIIIc), FGFR2b (FGFR2IIIb), FGFR2c (FGFR2IIIc), FGFR3b (FGFR3IIIb), FGFR3c (FGFR3IIIc), and FGFR4.
[0033] The full length FGF1 (GenBank Accession Nos: BC032697.1 and AAH32697.1) comprises a non-glycosylated 155 amino acid residue polypeptide (SEQ ID NO: 1). The mature FGF1 comprises 140 amino acid residues in length (SEQ ID NO: 2), which is resulted from the truncation of the first 15 residues from the N-terminus of the full length FGF1.
[0034] In some instances, FGF1 is expressed in metabolic tissues such as liver, kidney, muscle, and white adipose tissue (WAT). In particular, FGF1 is upregulated in WAT following a high-fat food intake. Each cell of the white adipose tissue, or white fat, comprises a single large droplet of fat with a nucleus encircling the droplet as a thin rim. In some instances, WAT is further grouped into visceral or gonadal WAT (gWAT) or inguinal white adipose tissue (iWAT). In some cases, FGF1 has been observed in the adipocyte fraction but not in the stromal vascular fraction (SVF) of gWAT and in the adipocyte fraction of iWAT.
[0035] In some cases, downregulation of FGF 1 post high-fat diet challenge has been correlated with diabetic and/or diabetic-related phenotypes in mice. Indeed, FGF1 knockout mice has been characterized by aberrant WAT expansion, impaired WAT vascularization, and rapid development of severe diabetes upon high-fat diet (HFD) challenge and defective WAT reduction upon HFD withdrawal (Jonker, et al, "A PPARy-FGFl axis is required for adaptive adipose remodeling and metabolic homeostasis," Nature 485(7398): 391-394 (2012)).
[0036] The nuclear receptor peroxisome proliferator-activated receptor γ (PPARy), also known as glitazone receptor or NR1C3, is the master adipocyte regulator. In some instances, there are two isoforms of PPARy, PPAR-γΙ, which is expressed in all tissues except muscle, and PPAR-y2, which is expressed in adipose tissue and the intestine. FGF1 is downstream of PPARy, and in some instances, is proposed to mediate a subset of PPARy-activated genes, and thereby modulates metabolic profile. In some cases, administration of FGF 1 has been shown to normalize blood glucose levels in obese, diabetic rodents, while chronic administration of FGF 1 has been shown to induce normoglycemia, insulin sensitization, and reduced hepatosteatosis (fatty liver) (Nies, et al,
"Fibroblast growth factor signaling in metabolic regulation," Frontiers in Endocrinology 6(193): 1-15 (2016)).
[0037] Fibroblast growth factor 2 (also known as basic fibroblast growth factor, bFGF, FGF2, or FGF-β), along with FGF1 forms the FGF1 subgroup. FGF2 is a 288 amino acid polypeptide that comprises about 5 different isoforms. The C-terminal portion of FGF2 shares about 48% sequence identity to FGF 1. Similar to FGF 1 , FGF2 is involved in cellular developmental processes such as proliferation, migration, differentiation, and survival. In some instances, FGF2 has been implicated in the pathogenesis of vascular dysfunction induced by hyperglycemia, such as vascular
hyperpermeability and hemodynamic change, increased aortic smooth muscle growth response to vascular injury, and the development of diabetic retinopathy, (see Stephan, C.C., et al., "Role for heparin-binding growth factors in glucose-induced vascular dysfunction," Diabetes 47: 1771-1778 (1998); and Lindner, et al., "Proliferation of smooth muscle cells after vascular injury is inhibited by an antibody against basic fibroblast growth factor," Proc. Natl . Acad. Sci. USA 88: 3739-3743 (1991))
[0038] In some instances, the mitogenic activity of FGF 1 is potentiated by heparan sulfate or a heparan sulfate analogue, leading to cellular developments such as proliferation, migration, and differentiation. In some cases, the mitogenic activity of FGF 1 has also been linked to increased tumorigenicity. Indeed, studies have shown that FGF1 stimulate the cell growth of breast cancer, prostate cancer, and bladder cancer. See, e.g., Fernig, et al., "Differential regulation of FGF-1 and -2 mitogenic activity is related to their kinetics of binding to heparan sulfate in MDA-MB-231 human breast cancer cells," Biochemical and Biophysical Research Comm. 267(3): 770-776 (2000);
Levesque, et al., "Effect of fibroblastic growth factors (FGF) on steroid UDP-glucuronosyltransferase expression and activity in the LNCaP cell line," J Steroid Biochem Mol Biol 64(1-2): 43-48 (1998); and Tomlinson, et al., "Altered splicing of FGFR1 is associated with high tumor grade and stage and leads to increased sensitivity to FGF1 in bladder cancer," Am J Pathol. 177(5): 2379-2386 (2010).
[0039] In some embodiments, disclosed herein are FGF mutants and methods of use in the treatment of a metabolic disease. In some embodiments, the FGF mutants exhibit low mitogenicity. In some cases, the FGF mutants further exhibit near wild-type FGFR interaction. In additional cases, the FGF mutants further induces and/or enhances insulin sensitivity in a subject. In some instances, the FGF mutant is an FGF1 mutant. In some instances, described herein are FGF1 mutants and methods of use in the treatment of a metabolic disease. In some cases, the FGF1 mutants exhibit low mitogenicity and near wild-type FGFR interaction, and further optionally induces and/or enhances insulin sensitivity in a subject.
[0040] In some embodiments, disclosed herein is an isolated and purified FGF polypeptide which comprises at least 80% sequence identity to SEQ ID NOs: 3-25917 and 25922-25974. In some embodiments, also disclosed herein is an isolated and purified FGF polypeptide comprising a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2. In additional embodiments, disclosed herein is an isolated and purified FGF polypeptide that comprises a modification at amino acid residue K10, Y15, S17, N18, H21, L26, P27, D28, R35, D36, R37, S38, D39, Q40, Q43, S47, Y55, E60, T61, Y64, D68, E81, E87, R88, E90, E91, N92, Y94, N95, K100, K101, H102, K105, Kl 13, SI 16, CI 17, Kl 18, P121, R122, H124, Q 127, or L133, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2.
[0041] In some embodiments, disclosed herein is a pharmaceutical composition, which comprises an isolated and purified FGF polypeptide, such as an isolated and purified FGF polypeptide which comprises at least 80% sequence identity to SEQ ID NOs: 3-25917 and 25922-25974; an isolated and purified FGF polypeptide comprising a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof; or an isolated and purified FGF polypeptide that comprises a modification at amino acid residue K10, Y15, S17, N18, H21, L26, P27, D28, R35, D36, R37, S38, D39, Q40, Q43, S47, Y55, E60, T61, Y64, D68, E81, E87, R88, E90, E91, N92, Y94, N95, K100, K101, H102, K105, Kl 13, S I 16, CI 17, Kl 18, P121, R122, H124, Q127, or L133, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2; and an excipient. [0042] In some embodiments, also disclosed herein is method of treating a metabolic disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an isolated and purified FGF polypeptide, such as an isolated and purified FGF polypeptide which comprises at least 80% sequence identity to SEQ ID NOs: 3-25917 and 25922- 25974; an isolated and purified FGF polypeptide comprising a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof; or an isolated and purified FGF polypeptide that comprises a modification at amino acid residue K10, Y15, S 17, N18, H21, L26, P27, D28, R35, D36, R37, S38, D39, Q40, Q43, S47, Y55, E60, T61, Y64, D68, E81, E87, R88, E90, E91, N92, Y94, N95, K100, K101, H102, K105, Kl 13, S 116, CI 17, K118, P121, R122, H124, Q127, or L133, wherein the numbering of the amino acid residue corresponds to SEQ ID NO: 2.
[0043] In additional embodiments, disclosed herein include polynucleic acid molecules that encode one or more of a FGF polypeptide described herein, and kits comprising an isolated and purified FGF polypeptide described herein or polynucleic acid molecule that encodes a FGF polypeptide described herein.
FGF Mutants
[0044] In some embodiments, disclosed herein are FGF mutants and methods of use in the treatment of a metabolic disease. In some instances, the FGF mutant is an FGFl or FGF2 mutant. In some instances, the FGF mutant is an FGF 1 mutant.
[0045] In some instances, the FGF mutant is a FGF polypeptide that comprises a truncation at the N-terminus, the C-terminus, or truncations at both termini. In some instances, the truncation is a truncation of about 1 to about 30 amino acid residues. In some instances, the truncation is a truncation of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more amino acid residues. In some cases, the truncation is a truncation of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more amino acid residues at the N-terminus. In some cases, the truncation is a truncation of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more amino acid residues at the C-terminus.
[0046] In some embodiments, the FGF mutant is an FGFl polypeptide that comprises 155 amino acid residues (or comprises the full length FGFl sequence; SEQ ID NO: 1). In some instances, the FGF mutant is an FGFl polypeptide that comprises 140 amino acid residues (or comprises the mature FGFl sequence; SEQ ID NO: 2). In some instances, the FGF mutant is a mature FGFl (140 amino acid residues) that further comprises a truncation at the N-terminus, the C-terminus, or truncations at both termini. In some instances, the truncation is a truncation of about 1 to about 30 amino acid residues. In some instances, the truncation is a truncation of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more amino acid residues. In some cases, the truncation is a truncation of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more amino acid residues at the N-terminus based on the mature FGFl polypeptide (SEQ ID NO: 2). In some cases, the truncation is a truncation of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more amino acid residues at the C-terminus based on the mature FGFl polypeptide (SEQ ID NO: 2).
[0047] In some embodiments, the FGF mutant is a FGF polypeptide that comprises about 1 to about 20 modifications. In some instances, the FGF mutant is a FGF polypeptide that comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 modifications. In some instances, the FGF mutant is an FGFl polypeptide with about 1 to about 20 modifications. In some instances, the FGF mutant is a FGF2 polypeptide with about 1 to about 20 modifications.
[0048] In some embodiments, the FGF mutant is an FGFl polypeptide that comprises about 1 to about 20 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 1 modification. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 2 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 3 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 4 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 5 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 6 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 7 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 8 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 9 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 10 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 11 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 12 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 13 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 14 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 15 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 16 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 17 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 18 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 19 modifications. In some instances, the FGF mutant is an FGFl polypeptide that comprises about 20 modifications. In some cases, a modification comprises an insertion, a deletion, or a mutation. In some instances, the mutation is a missense mutation or a nonsense mutation. In some cases, the insertion comprises a duplication or one or more amino acid residues or a repeat expansion of one or more amino acid residues. [0049] In some embodiments, the FGF mutant is a truncated FGF 1 polypeptide that further comprises one or more modifications described herein. In some instances, the FGF mutant is a N- terminus truncated FGFl polypeptide that further comprises about 1 to about 20 modifications. In some cases, the FGF mutant is a C-terminus truncated FGFl polypeptide that further comprises about 1 to about 20 modifications. In additional cases, the FGF mutant is a N-terminus and a C-terminus truncated FGFl polypeptide that further comprises about 1 to about 20 modifications. In some instances, the truncation is a truncation of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more amino acid residues. In some cases, the FGFl polypeptide comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 modifications.
[0050] In some embodiments, the modification comprises a mutation to a natural or unnatural amino acid residue. For example, in some cases, the modification comprises a mutation of an amino acid residue to an alternative natural amino acid residue, for example a mutation of an arginine to an alanine. In some instances, the modification comprises a mutation of an amino acid residue to an unnatural amino acid residue, for example, a mutation of a cysteine to a seleno-L-cystine.
[0051] As used herein, an amino acid residue is a molecule containing both an amino group and a carboxyl group. Suitable amino acids for use in a FGF mutant described herein include, without limitation, both the D- and L-isomers of the naturally-occurring amino acids, as well as non-naturally occurring amino acids prepared by organic synthesis or other metabolic routes. In some instances, an amino acid is an a-amino acid, β-amino acid, natural amino acid, non-natural amino acid, or amino acid analog. A naturally occurring amino acid comprises any one of the twenty amino acids commonly found in peptides synthesized in nature, and known by the one letter abbreviations A, R, N, C, D, Q, E, G, H, I, L, K, M, F, P, S, T, W, Y and V.
[0052] Exemplary unnatural amino acid residues comprise, for example, amino acid analogs such as β-amino acid analogs; racemic analogs; or analogs of amino acid residue alanine, valine, glycine, leucine, arginine, lysine, aspartic acid, glutamic acid, cysteine, methionine, tyrosine, phenylalanine, tryptophane, serine, threonine, or proline. Exemplary β-amino acid analogs include, but are not limited to, cyclic β-amino acid analogs, β-alanine, (R)^-phenylalanine, (R)- 1,2,3, 4-tetrahydro- isoquinoline-3-acetic acid, (R)-3-amino-4-(l-naphthyl)-butyric acid, (R)-3-amino-4-(2,4- dichlorophenyl)butyric acid, (R)-3-amino-4-(2-chlorophenyl)-butyric acid, (R)-3-amino-4-(2- cyanophenyl)-butyric acid, (R)-3-amino-4-(2-fluorophenyl)-butyric acid, (R)-3-amino-4-(2-furyl)- butyric acid, (R)-3-amino-4-(2-methylphenyl)-butyric acid, (R)-3-amino-4-(2-naphthyl)-butyric acid, (R)-3-amino-4-(2-thienyl)-butyric acid, (R)-3-amino-4-(2-trifluoromethylphenyl)-butyric acid, (R)-3- amino-4-(3,4-dichlorophenyl)butyric acid, (R)-3-amino-4-(3,4-difluorophenyl)butyric acid, (R)-3- amino-4-(3-benzothienyl)-butyric acid, (R)-3-amino-4-(3-chlorophenyl)-butyric acid, (R)-3-amino-4- (3-cyanophenyl)-butyric acid, (R)-3-amino-4-(3-fluorophenyl)-butyric acid, (R)-3-amino-4-(3- methylphenyl) -butyric acid, (R)-3-amino-4-(3-pyridyl)-butyric acid, (R)-3-amino-4-(3-thienyl)- butyric acid, (R)-3-amino-4-(3-trifluoromethylphenyl)-butyric acid, (R)-3-amino-4-(4-bromophenyl)- butyric acid, (R)-3-amino-4-(4-chlorophenyl)-butyric acid, (R)-3-amino-4-(4-cyanophenyl)-butyric acid, (R)-3-amino-4-(4-fluorophenyl)-butyric acid, (R)-3-amino-4-(4-iodophenyl)-butyric acid, (R)-3- amino-4-(4-methylphenyl)-butyric acid, (R)-3-amino-4-(4-nitrophenyl)-butyric acid, (R)-3-amino-4- (4-pyridyl)-butyric acid, (R)-3-amino-4-(4-trifluoromethylphenyl)-butyric acid, (R)-3-amino-4- pentafluoro-phenylbutyric acid, (R)-3-amino-5-hexenoic acid, (R)-3-amino-5-hexynoic acid, (R)-3- amino-5-phenylpentanoic acid, (R)-3-amino-6-phenyl-5-hexenoic acid, (S)- 1,2,3, 4-tetrahydro- isoquinoline-3-acetic acid, (S)-3-amino-4-(l-naphthyl)-butyric acid, (S)-3-amino-4-(2,4- dichlorophenyl)butyric acid, (S)-3-amino-4-(2-chlorophenyl)-butyric acid, (S)-3-amino-4-(2- cyanophenyl)-butyric acid, (S)-3-amino-4-(2-fluorophenyl)-butyric acid, (S)-3-amino-4-(2-furyl)- butyric acid, (S)-3-amino-4-(2-methylphenyl)-butyric acid, (S)-3-amino-4-(2-naphthyl)-butyric acid, (S)-3-amino-4-(2-thienyl)-butyric acid, (S)-3-amino-4-(2-trifluoromethylphenyl)-butyric acid, (S)-3- amino-4-(3,4-dichlorophenyl)butyric acid, (S)-3-amino-4-(3,4-difluorophenyl)butyric acid, (S)-3- amino-4-(3-benzothienyl)-butyric acid, (S)-3-amino-4-(3-chlorophenyl)-butyric acid, (S)-3-amino-4- (3-cyanophenyl)-butyric acid, (S)-3-amino-4-(3-fluorophenyl)-butyric acid, (S)-3-amino-4-(3- methylphenyl)-butyric acid, (S)-3-amino-4-(3-pyridyl)-butyric acid, (S)-3-amino-4-(3-thienyl)-butyric acid, (S)-3-amino-4-(3-trifluoromethylphenyl)-butyric acid, (S)-3-amino-4-(4-bromophenyl)-butyric acid, (S)-3-amino-4-(4-chlorophenyl) butyric acid, (S)-3-amino-4-(4-cyanophenyl)-butyric acid, (S)- 3-amino-4-(4-fluorophenyl) butyric acid, (S)-3-amino-4-(4-iodophenyl)-butyric acid, (S)-3-amino-4- (4-methylphenyl)-butyric acid, (S)-3-amino-4-(4-nitrophenyl)-butyric acid, (S)-3-amino-4-(4- pyridyl)-butyric acid, (S)-3-amino-4-(4-trifluoromethylphenyl)-butyric acid, (S)-3-amino-4- pentafluoro-phenylbutyric acid, (S)-3-amino-5-hexenoic acid, (S)-3-amino-5-hexynoic acid, (S)-3- amino-5-phenylpentanoic acid, (S)-3-amino-6-phenyl-5-hexenoic acid, 1,2,5, 6-tetrahydropyridine-3- carboxylic acid, 1,2,5, 6-tetrahydropyridine-4-carboxylic acid, 3-amino-3-(2-chlorophenyl)-propionic acid, 3-amino-3-(2-thienyl)-propionic acid, 3-amino-3-(3-bromophenyl)-propionic acid, 3-amino-3- (4-chlorophenyl)-propionic acid, 3-amino-3-(4-methoxyphenyl)-propionic acid, 3-amino-4,4,4- trifluoro-butyric acid, 3-aminoadipic acid, D- -phenylalanine, β-leucine, L- -homoalanine, L-β- homoaspartic acid γ-benzyl ester, L- -homoglutamic acid δ-benzyl ester, L- -homoisoleucine, L-β- homoleucine, L- -homomethionine, L- -homophenylalanine, L- -homoproline, L- -homotryptophan, L- -homovaline, L-Nco-benzyloxycarbonyl- -homolysine, Nco-L- -homoarginine, O-benzyl-L-β- homohydroxyproline, O-benzyl-L- -homoserine, O-benzyl-L- -homothreonine, O-benzyl-L-β- homotyrosine, y-trityl-L- -homoasparagine, (R)- -phenylalanine, L- -homoaspartic acid γ-t-butyl ester, L- -homoglutamic acid δ-t-butyl ester, L-Nco- -homolysine, N5-trityl-L- -homoglutamine, Νω- 2,2,4,6,7-pentamethyl-dihydrobenzofuran-5-sulfonyl-L- -homoarginine, O-t-butyl-L-β- homohydroxy-proline, O-t-butyl-L- -homoserine, O-t-butyl-L- -homothreonine, O-t-butyl-L-β- homotyrosine, 2-aminocyclopentane carboxylic acid, and 2-aminocyclohexane carboxylic acid. [0053] In some instances, unnatural amino acid residues comprise a racemic mixture of amino acid analogs. For example, in some instances, the D isomer of the amino acid analog is used. In some cases, the L isomer of the amino acid analog is used. In some instances, the amino acid analog comprises chiral centers that are in the R or S configuration. Sometimes, the amino group(s) of a β-amino acid analog is substituted with a protecting group, e.g., tert-butyloxycarbonyl (BOC group), 9- fluorenylmethyloxycarbonyl (FMOC), tosyl, and the like. Sometimes, the carboxylic acid functional group of a β-amino acid analog is protected, e.g., as its ester derivative. In some cases, the salt of the amino acid analog is used.
[0054] In some cases, unnatural amino acid residues comprise analogs of amino acid residue alanine, valine, glycine, leucine, arginine, lysine, aspartic acid, glutamic acid, cysteine, methionine, tyrosine, phenylalanine, tryptophane, serine, threonine, or proline. Exemplary amino acid analogs of alanine, valine, glycine, and leucine include, but are not limited to, a-methoxyglycine, a-allyl-L- alanine, a-aminoisobutyric acid, a-methyl-leucine, -(l-naphthyl)-D-alanine, P-(l-naphthyl)-L- alanine, P-(2-naphthyl)-D-alanine, P-(2-naphthyl)-L-alanine, -(2-pyridyl)-D-alanine, -(2-pyridyl)-L- alanine, -(2-thienyl)-D-alanine, -(2-thienyl)-L-alanine, -(3-benzothienyl)-D-alanine, β-(3- benzothienyl)-L-alanine, -(3-pyridyl)-D-alanine, -(3-pyridyl)-L-alanine, -(4-pyridyl)-D-alanine, β- (4-pyridyl)-L-alanine, β-chloro-L-alanine, β-cyano-L-alanin, β-cyclohexyl-D-alanine, β-cyclohexyl- L-alanine, β-cyclopenten- l-yl -alanine, β-cyclopentyl-alanine, β-cyclopropyl-L-Ala- OH.dicyclohexylammonium salt, β-t-butyl-D-alanine, β-t-butyl-L-alanine, γ-aminobutyric acid, L- α,β-diaminopropionic acid, 2,4-dinitro-phenylglycine, 2,5-dihydro-D-phenylglycine, 2-amino-4,4,4- trifluorobutyric acid, 2-fluoro-phenylglycine, 3-amino-4,4,4-trifluoro-butyric acid, 3-fluoro-valine, 4,4,4-trifluoro-valine, 4,5-dehydro-L-leu-OH.dicyclohexylammonium salt, 4-fluoro-D-phenylglycine, 4-fluoro-L-phenylglycine, 4-hydroxy-D-phenylglycine, 5,5,5-trifluoro-leucine, 6-aminohexanoic acid, cyclopentyl-D-Gly-OH.dicyclohexylammonium salt, cyclopentyl-Gly-OH.dicyclohexylammonium salt, D-a^-diaminopropionic acid, D-a-aminobutyric acid, D-a-t-butylglycine, D-(2-thienyl)glycine, D-(3-thienyl)glycine, D-2-aminocaproic acid, D-2-indanylglycine, D-allylglycine- dicyclohexylammonium salt, D-cyclohexylglycine, D-norvaline, D-phenylglycine, β-aminobutyric acid, β-aminoisobutyric acid, (2-bromophenyl)glycine, (2-methoxyphenyl)glycine, (2- methylphenyl)glycine, (2-thiazoyl)glycine, (2-thienyl)glycine, 2-amino-3-(dimethylamino)-propionic acid, L-a^-diaminopropionic acid, L-a-aminobutyric acid, L-a-t-butylglycine, L-(3-thienyl)glycine, L-2-amino-3-(dimethylamino)-propionic acid, L-2-aminocaproic acid dicyclohexyl -ammonium salt, L-2-indanylglycine, L-allylglycine. dicyclohexyl ammonium salt, L-cyclohexylglycine, L- phenylglycine, L-propargylglycine, L-norvaline, N-a-aminomethyl-L-alanine, D-a,y-diaminobutyric acid, L-a,y-diaminobutyric acid, β-cyclopropyl-L-alanine, ( ^-(2,4-dinitrophenyl))-L-a^- diaminopropionic acid, (Ν-β- 1 -(4,4-dimethyl-2,6-dioxocyclohex- 1 -ylidene)ethyl)-D-a^- diaminopropionic acid, (Ν-β- 1 -(4,4-dimethyl-2,6-dioxocyclohex- 1 -ylidene)ethyl)-L-a^- diaminopropionic acid, (N-β-4-methyltrityl)-L-α,β-diaminopropionic acid, (N^-allyloxycarbonyl)-L- α,β-diaminopropionic acid, (N-Y-l-(4,4-dimethyl-2,6-dioxocyclohex- l-ylidene)ethyl)-D-a,y- diaminobutyric acid, (Ν-γ- 1 -(4,4-dimethyl-2,6-dioxocyclohex- 1 -ylidene)ethyl)-L-a,y-diaminobutyric acid, (N-Y-4-methyltrityl)-D-a,y-diaminobutyric acid, (N-Y-4-methyltrityl)-L-a,Y-diaminobutyric acid, (N-y-allyloxycarbonyl)-L-a,Y-diaminobutyric acid, D-a,y-diaminobutyric acid, 4,5-dehydro-L-leucine, cyclopentyl-D-Gly-OH, cyclopentyl-Gly-OH, D-allylglycine, D-homocyclohexylalanine, L-l- pyrenylalanine, L-2-aminocaproic acid, L-allylglycine, L-homocyclohexylalanine, and N-(2-hydroxy- 4-methoxy-Bzl)-Gly-OH.
[0055] Exemplary amino acid analogs of arginine and lysine include, but are not limited to, citrulline, L-2-amino-3-guanidinopropionic acid, L-2-amino-3-ureidopropionic acid, L-citrulline, Lys(Me)2-OH, Lys(N3)— OH, Νδ-benzyloxycarbonyl-L-ornithine, Νω-nitro-D-arginine, Nco-nitro-L- arginine, a-methyl -ornithine, 2,6-diaminoheptanedioic acid, L-ornithine, (N5- l-(4,4-dimethyl-2,6- dioxo-cyclohex- 1 -ylidene)ethyl)-D-ornithine, (Νδ- 1 -(4,4-dimethyl-2,6-dioxo-cyclohex- 1 - ylidene)ethyl)-L-ornithine, (N5-4-methyltrityl)-D-ornithine, (N5-4-methyltrityl)-L-ornithine, D- ornithine, L-ornithine, Arg(Me)(Pbf)-OH, Arg(Me)2-OH (asymmetrical), Arg(Me)2-OH
(symmetrical), Lys(ivDde)-OH, Lys(Me)2-OH.HCl, Lys(Me3)-OH chloride, Νω-nitro-D-arginine, and Νω-nitro-L-arginine.
[0056] Exemplary amino acid analogs of aspartic and glutamic acids include, but are not limited to, a-methyl-D-aspartic acid, a-methyl-glutamic acid, a-methyl-L-aspartic acid, γ-methylene-glutamic acid, (N-Y-ethyl)-L-glutamine, [N-a-(4-aminobenzoyl)]-L-glutamic acid, 2,6-diaminopimelic acid, L- a-aminosuberic acid, D-2-aminoadipic acid, D-a-aminosuberic acid, a-aminopimelic acid, iminodiacetic acid, L-2-aminoadipic acid, threo- -methyl-aspartic acid, γ-carboxy-D-glutamic acid γ,γ-di-t-butyl ester, γ-carboxy-L-glutamic acid γ,γ-di-t-butyl ester, Glu(OAll)-OH, L-Asu(OtBu)— OH, and pyroglutamic acid.
[0057] Exemplary amino acid analogs of cysteine and methionine include, but are not limited to, Cys(farnesyl)-OH, Cys(farnesyl)-OMe, a-methyl-methionine, Cys(2-hydroxyethyl)-OH, Cys(3- aminopropyl)-OH, 2-amino-4-(ethylthio)butyric acid, buthionine, buthioninesulfoximine, ethionine, methionine methylsulfonium chloride, selenomethionine, cysteic acid, [2-(4-pyridyl)ethyl]-DL- penicillamine, [2-(4-pyridyl)ethyl]-L-cysteine, 4-methoxybenzyl-D-penicillamine, 4-methoxybenzyl- L-penicillamine, 4-methylbenzyl-D-penicillamine, 4-methylbenzyl-L-penicillamine, benzyl-D- cysteine, benzyl-L-cysteine, benzyl-DL-homocysteine, carbamoyl-L-cysteine, carboxyethyl-L- cysteine, carboxymethyl-L-cysteine, diphenylmethyl-L-cysteine, ethyl-L-cysteine, methyl-L-cysteine, t-butyl-D-cysteine, trityl-L-homocysteine, trityl-D-penicillamine, cystathionine, homocystine, L- homocystine, (2-aminoethyl)-L-cysteine, seleno-L-cystine, cystathionine, Cys(StBu)— OH, and acetamidomethyl-D-penicillamine.
[0058] Exemplary amino acid analogs of phenylalanine and tyrosine include, but are not limited to, β-methyl-phenylalanine, β-hydroxyphenylalanine, a-methyl-3-methoxy-DL-phenylalanine, a-methyl- D-phenylalanine, α-methyl-L-phenylalanine, l,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, 2,4- dichloro-phenylalanine, 2-(trifluoromethyl)-D-phenylalanine, 2-(trifluoromethyl)-L-phenylalanine, 2- bromo-D-phenylalanine, 2-bromo-L-phenylalanine, 2-chloro-D-phenylalanine, 2-chloro-L- phenylalanine, 2-cyano-D-phenylalanine, 2-cyano-L-phenylalanine, 2-fluoro-D-phenylalanine, 2- fluoro-L-phenylalanine, 2-methyl-D-phenylalanine, 2-methyl-L-phenylalanine, 2-nitro-D- phenylalanine, 2-nitro-L-phenylalanine, 2,4,5-trihydroxy-phenylalanine, 3,4,5-trifluoro-D- phenylalanine, 3,4,5-trifluoro-L-phenylalanine, 3,4-dichloro-D-phenylalanine, 3,4-dichloro-L- phenylalanine, 3,4-difluoro-D-phenylalanine, 3,4-difluoro-L-phenylalanine, 3,4-dihydroxy-L- phenylalanine, 3,4-dimethoxy-L-phenylalanine, 3,5,3'-triiodo-L-thyronine, 3,5-diiodo-D-tyrosine, 3,5- diiodo-L-tyrosine, 3,5-diiodo-L-thyronine, 3-(trifluoromethyl)-D-phenylalanine, 3-(trifluoromethyl)- L-phenylalanine, 3-amino-L-tyrosine, 3-bromo-D-phenylalanine, 3-bromo-L-phenylalanine, 3-chloro- D-phenylalanine, 3-chloro-L-phenylalanine, 3-chloro-L-tyrosine, 3-cyano-D-phenylalanine, 3-cyano- L-phenylalanine, 3-fluoro-D-phenylalanine, 3-fluoro-L-phenylalanine, 3-fluoro-tyrosine, 3-iodo-D- phenylalanine, 3-iodo-L-phenylalanine, 3-iodo-L-tyrosine, 3-methoxy-L-tyrosine, 3-methyl-D- phenylalanine, 3 -methyl -L-phenylalanine, 3-nitro-D-phenylalanine, 3-nitro-L-phenylalanine, 3-nitro- L-tyrosine, 4-(trifluoromethyl)-D-phenylalanine, 4-(trifluoromethyl)-L-phenylalanine, 4-amino-D- phenylalanine, 4-amino-L-phenylalanine, 4-benzoyl-D-phenylalanine, 4-benzoyl-L-phenylalanine, 4- bis(2-chloroethyl)amino-L-phenylalanine, 4-bromo-D-phenylalanine, 4-bromo-L-phenylalanine, 4- chloro-D-phenylalanine, 4-chloro-L-phenylalanine, 4-cyano-D-phenylalanine, 4-cyano-L- phenylalanine, 4-fluoro-D-phenylalanine, 4-fluoro-L-phenylalanine, 4-iodo-D-phenylalanine, 4-iodo- L-phenylalanine, homophenylalanine, thyroxine, 3,3-diphenylalanine, thyronine, ethyl-tyrosine, and methyl -tyrosine.
[0059] Exemplary amino acid analogs of proline include 3,4-dehydro-proline, 4-fluoro-proline, cis- 4-hydroxy-proline, thiazolidine-2-carboxylic acid, and trans-4-fluoro-proline.
[0060] Exemplary amino acid analogs of serine and threonine include 3-amino-2-hydroxy-5- methylhexanoic acid, 2-amino-3 -hydroxy -4-methylpentanoic acid, 2-amino-3-ethoxybutanoic acid, 2- amino-3-methoxybutanoic acid, 4-amino-3-hydroxy-6-methylheptanoic acid, 2-amino-3- benzyloxypropionic acid, 2-amino-3-benzyloxypropionic acid, 2-amino-3-ethoxypropionic acid, 4- amino-3-hydroxybutanoic acid, and a-methylserine.
[0061] Exemplary amino acid analogs of tryptophan include, but are not limited to, a-methyl- tryptophan, -(3-benzothienyl)-D-alanine, -(3-benzothienyl)-L-alanine, 1 -methyl -tryptophan, 4- methyl-tryptophan, 5 -benzyloxy -tryptophan, 5 -bromo -tryptophan, 5-chloro-tryptophan, 5-fluoro- tryptophan, 5-hydroxy-tryptophan, 5-hydroxy-L-tryptophan, 5 -methoxy -tryptophan, 5-methoxy-L- tryptophan, 5-methyl-tryptophan, 6-bromo-tryptophan, 6-chloro-D-tryptophan, 6-chloro-tryptophan, 6-fluoro-tryptophan, 6-methyl-tryptophan, 7-benzyloxy-tryptophan, 7-bromo-tryptophan, 7-methyl- tryptophan, D-l,2,3,4-tetrahydro-norharman-3-carboxylic acid, 6-methoxy-l,2,3,4- tetrahydronorharman-l-carboxylic acid, 7-azatryptophan, L- 1,2,3, 4-tetrahydro-norharman-3- carboxylic acid, 5 -methoxy-2 -methyl -tryptophan, and 6-chloro-L-tryptophan.
[0062] In some instances, the modification comprises a mutation to a hydrophobic or nonpolar amino acid residue. In some cases, a hydrophobic or nonpolar amino acid includes small hydrophobic amino acids and large hydrophobic amino acids. Exemplary small hydrophobic amino acids include glycine, alanine, proline, and analogs thereof. Exemplary large hydrophobic amino acids include valine, leucine, isoleucine, phenylalanine, methionine, tryptophan, and analogs thereof. In some instances, a FGF mutant described herein comprises a mutation to a small hydrophobic amino acid (e.g., a mutation to glycine, alanine, proline, and analogs thereof). In some instances, a FGF mutant described herein comprises a mutation to a large hydrophobic amino acid (e.g., a mutation to valine, leucine, isoleucine, phenylalanine, methionine, tryptophan, and analogs thereof).
[0063] In some instances, the modification comprises a mutation to a polar amino acid residue. In some cases, the polar amino acid comprises serine, threonine, asparagine, glutamine, cysteine, tyrosine, and analogs thereof. In some instances, a FGF mutant described herein comprises a mutation to a polar amino acid residue (e.g., a mutation to serine, threonine, asparagine, glutamine, cysteine, tyrosine, and analogs thereof).
[0064] In additional instances, the modification comprises a mutation to a charged amino acid residue. In some cases, charged amino acids include lysine, arginine, histidine, aspartate, glutamate, or analog thereof. In some instances, a FGF mutant described herein comprises a mutation to a charged amino acid residue (e.g., a mutation to lysine, arginine, histidine, aspartate, glutamate, or analog thereof).
[0065] In some embodiments, a FGF mutant described herein comprise a modification to a nonessential amino acid. In some instances, a non-essential amino acid residue is a residue that is altered from the wild-type sequence of a polypeptide without abolishing or substantially altering its essential biological or biochemical activity (e.g., receptor binding or activation). In some cases, a FGF mutant provided herein comprises an essential amino acid. In some cases, an essential amino acid residue is a residue that, when altered from the wild-type sequence of the polypeptide, results in abolishing or substantially abolishing the polypeptide's essential biological or biochemical activity.
[0066] In some embodiments, a FGF mutant described herein comprises a conservative amino acid substitution. In some cases, a conservative amino acid substitution is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include, for example, amino acids with basic side chains (e.g., K, R, H), acidic side chains (e.g., D, E), uncharged polar side chains (e.g., G, N, Q, S, T, Y, C), nonpolar side chains (e.g., A, V, L, I, P, F, M, W), beta-branched side chains (e.g., T, V, I) and aromatic side chains (e.g., Y, F, W, H). Thus, a predicted nonessential amino acid residue in a polypeptide, for example, is replaced with another amino acid residue from the same side chain family. Other examples of acceptable substitutions include substitutions based on isosteric considerations (e.g., norleucine for methionine) or other properties (e.g., 2-thienylalanine for phenylalanine, or 6-Cl-tryptophan for tryptophan).
[0067] In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at at least one amino acid position: 30, 34, 41, 42, 43, 48, 53, 57, 63, 78, 79, 84, 89, 96, or 128, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at at least one amino acid position: 30, 34, 48, 53, 57, 63, 78, 79, 84, 89, 96, or 128, according to the amino acid residue position of SEQ ID NO: 2. In some instances, the FGF mutant is an FGFl polypeptide that comprises a modification at at least one amino acid position: 30, 34, 48, 53, 57, 63, 78, 79, 89, 96, or 128, according to the amino acid residue position of SEQ ID NO: 2. In some cases, the FGF mutant is an FGFl polypeptide that comprises a modification at at least amino acid position 84, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 30, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 34, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 41, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 42, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 43, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 48, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 53, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 57, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 63, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 78, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 79, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 84, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 89, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 96, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 128, according to the amino acid residue position of SEQ ID NO: 2.
[0068] In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at at least one amino acid position: 10, 11, 12, 13, 15, 17, 18, 21, 26, 27, 28, 35, 36, 37, 38, 39, 40, 43, 44, 46, 47, 55, 60, 61, 64, 68, 81, 87, 88, 90, 91, 92, 93, 94, 95, 100, 101, 102, 105, 108, 113, 116, 117, 118, 121, 122, 124, 127, 133, or 134, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 10, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 11, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 12, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid positions 13, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid positions 15, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid positions 17, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid positions 18, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 21, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 26, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 27, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 28, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid positions 35, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid positions 36, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid positions 37, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 38, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 39, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 40, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 43, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 44, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 46, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 47, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 55, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 60, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 61, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 64, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 68, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 81, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 87, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 88, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 90, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 91, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 92, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 93, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 94, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 95, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 100, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 101, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 102, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 105, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 108, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 113, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid positions 116, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid positions 117, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid positions 118, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 121, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 122, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 124, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 127, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 133, according to the amino acid residue position of SEQ ID NO: 2. In some embodiments, the FGF mutant is an FGFl polypeptide that comprises a modification at amino acid position 134, according to the amino acid residue position of SEQ ID NO: 2.
[0069] In some embodiments, an isolated and purified FGF polypeptide described herein comprises a modification at amino acid residue T30, T34, H41, 142, Q43, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof, according to the amino acid residue numbering of SEQ ID NO: 2. In some embodiments, an isolated and purified FGF polypeptide described herein comprises a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof, according to the amino acid residue numbering of SEQ ID NO: 2. In some instances, an isolated and purified FGF polypeptide described herein comprises a modification at amino acid residue T30, T34, A48, E53, K57, Q63, T78, P79, L89, T96, K128, or a combination thereof, according to the amino acid residue numbering of SEQ ID NO: 2. In some cases, an isolated and purified FGF polypeptide described herein comprises a modification at amino acid residue L84, according to the amino acid residue numbering of SEQ ID NO: 2.
[0070] In some instances, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to a non-polar residue. In other cases, the modification comprises a mutation to a charged residue. In some instances, the modification comprises a mutation to alanine.
[0071] In some instances, the isolated and purified FGF polypeptide comprising a modification at amino acid residue T30, T34, H41, 142, Q43, A48, E53, K57, Q63, T78, P79, L84, L89, T96, K128, or a combination thereof further comprises at least one additional modification. In some cases, the at least one additional modification comprises an insertion, a deletion, or a mutation. In some cases, the mutation comprises a mutation to a polar residue, a non-polar residue, or a charged residue. In additional cases, the isolated and purified FGF polypeptide further comprises a truncation. In additional cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
[0072] In some embodiments, the isolated and purified FGF polypeptide comprises a modification of T30A, T34A, A48G, E53A, K57A, K57E, Q63A, T78A, P79A, L84A, L89A, T96A, K128E, or a combination thereof. In some instances, the isolated and purified FGF polypeptide comprises T30A modification. In some instances, the isolated and purified FGF polypeptide comprises T34A modification. In some instances, the isolated and purified FGF polypeptide comprises A48G modification. In some instances, the isolated and purified FGF polypeptide comprises E53A modification. In some instances, the isolated and purified FGF polypeptide comprises K57A modification. In some instances, the isolated and purified FGF polypeptide comprises K57E modification. In some instances, the isolated and purified FGF polypeptide comprises Q63A modification. In some instances, the isolated and purified FGF polypeptide comprises T78A modification. In some instances, the isolated and purified FGF polypeptide comprises P79A modification. In some instances, the isolated and purified FGF polypeptide comprises L84A modification. In some instances, the isolated and purified FGF polypeptide comprises L89A modification. In some instances, the isolated and purified FGF polypeptide comprises T96A modification. In some instances, the isolated and purified FGF polypeptide comprises K128E modification.
[0073] In some embodiments, an isolated and purified FGF polypeptide described herein comprises a modification at amino acid residue K10, Y15, S17, N18, H21, L26, P27, D28, R35, D36, R37, S38, D39, Q40, Q43, S47, Y55, E60, T61, Y64, D68, E81, E87, R88, E90, E91, N92, Y94, N95, K100, K101, H102, K105, Kl 13, SI 16, CI 17, Kl 18, P121, R122, H124, Q127, L133, or a combination thereof, according to the amino acid residue numbering of SEQ ID NO: 2. In some cases, the mutation comprises a mutation to a polar residue, a non-polar residue, or a charged residue. In additional cases, the isolated and purified FGF polypeptide further comprises a truncation. In additional cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
[0074] In some instances, the isolated and purified FGF polypeptide comprising a modification at amino acid residue K10, Y15, S17, N18, H21, L26, P27, D28, R35, D36, R37, S38, D39, Q40, Q43, S47, Y55, E60, T61, Y64, D68, E81, E87, R88, E90, E91, N92, Y94, N95, K100, K101, H102, K105, K113, SI 16, CI 17, K118, P121, R122, H124, Q127, L133, or a combination thereof further comprises at least one additional modification. In some cases, the at least one additional modification comprises an insertion, a deletion, or a mutation. In some cases, the mutation comprises a mutation to a polar residue, a non-polar residue, or a charged residue. In additional cases, the isolated and purified FGF polypeptide further comprises a truncation. In additional cases, the truncation is an N- terminal truncation, a C-terminal truncation, or a combination thereof.
[0075] In some embodiments, an isolated and purified FGF polypeptide described herein comprises a modification at amino acid residue R35, according to the amino acid residue numbering of SEQ ID NO: 2. In some instances, the modification comprises a mutation to a polar residue. In some instances, the modification comprises a mutation to a non-polar residue. In some instances, the modification comprises a mutation to a different charged residue. In some cases, an isolated and purified FGF polypeptide described herein comprises modification: R35D, R35F, R35G, R35I, R35L, R35M, R35P, R35S, R35V, or R35E. In some instances, the isolated and purified FGF polypeptide further comprises at least one additional modification. In some cases, the at least one additional modification comprises an insertion, a deletion, or a mutation. In some cases, the mutation comprises a mutation to a polar residue, a non-polar residue, or a charged residue. In additional cases, the isolated and purified FGF polypeptide further comprises a truncation. In additional cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
[0076] In some embodiments, an isolated and purified FGF polypeptide described herein comprises a modification at amino acid residue SI 16, according to the amino acid residue numbering of SEQ ID NO: 2. In some instances, the modification comprises a mutation to a different polar residue. In some instances, the modification comprises a mutation to a non-polar residue. In some instances, the modification comprises a mutation to a charged residue. In some cases, an isolated and purified FGF polypeptide described herein comprises modification: SI 16F, S I 16G, S I 16N, SI 16W, or SI 16Y. In some instances, the isolated and purified FGF polypeptide further comprises at least one additional modification. In some cases, the at least one additional modification comprises an insertion, a deletion, or a mutation. In some cases, the mutation comprises a mutation to a polar residue, a non- polar residue, or a charged residue. In additional cases, the isolated and purified FGF polypeptide further comprises a truncation. In additional cases, the truncation is an N-terminal truncation, a C- terminal truncation, or a combination thereof. [0077] In some embodiments, an isolated and purified FGF polypeptide described herein comprises a modification at amino acid residue Kl 18, according to the amino acid residue numbering of SEQ ID NO: 2. In some instances, the modification comprises a mutation to a polar residue. In some instances, the modification comprises a mutation to a non-polar residue. In some instances, the modification comprises a mutation to a different charged residue. In some cases, an isolated and purified FGF polypeptide described herein comprises modification: Kl 18H, Kl 18R, Kl 18S, or Kl 18W. In some instances, the isolated and purified FGF polypeptide further comprises at least one additional modification. In some cases, the at least one additional modification comprises an insertion, a deletion, or a mutation. In some cases, the mutation comprises a mutation to a polar residue, a non-polar residue, or a charged residue. In additional cases, the isolated and purified FGF polypeptide further comprises a truncation. In additional cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
[0078] In some embodiments, described herein is a FGF polypeptide comprising one or more modified amino acids at Group 1 amino acid positions: 27, 35, 36, 37, 55, 57, 61, 81, 87, 88, 92, 96, 101, 102, and 133; and one or more modified amino acids at Group 2 amino acid positions: 26, 30, 39, 43, 64, 68, 77, 80, 82, 84, 86, 105, 120, 127, and 128, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the Group 1 amino acid positions are further subdivided into Group 1A: 35, 36, and 37; Group IB: 87, 88, 92, 96, and 133; and Group 1C: 27, 55, 57, 61, 81, 101, and 102. In some instances, the FGF polypeptide comprises one or more modified amino acids at Group 1A amino acid positions: 35, 36, and 37; and one or more modified amino acids at Group 2 amino acid positions: 26, 30, 39, 43, 64, 68, 77, 80, 82, 84, 86, 105, 120, 127, and 128, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises one or more modified amino acids at Group IB amino acid positions: 87, 88, 92, 96, and 133; and one or more modified amino acids at Group 2 amino acid positions: 26, 30, 39, 43, 64, 68, 77, 80, 82, 84, 86, 105, 120, 127, and 128, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises one or more modified amino acids at Group 1C amino acid positions: 27, 55, 57, 61, 81, 101, and 102; and one or more modified amino acids at Group 2 amino acid positions: 26, 30, 39, 43, 64, 68, 77, 80, 82, 84, 86, 105, 120, 127, and 128, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide is an FGF1 polypeptide.
[0079] In some embodiments, a FGF polypeptide described herein comprises one or more modified amino acids selected from Group 1 amino acid residues: P27, R35, D36, R37, Y55, K57, T61, E81, E87, R88, N92, T96, K101, H102, and L133; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the Group 1 modified amino acids are further divided into Group 1A amino acids: R35, D36, and R37; Group IB amino acids: E87, R88, N92, T96, and L133; and Group IC amino acids: P27, Y55, K57, T61, E81, K101, and H102. In some instances, the FGF polypeptide comprises one or more modified amino acids selected from Group 1A amino acid residues: R35, D36, and R37; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises one or more modified amino acids selected from Group IB amino acid residues: E87, R88, N92, T96, and L133; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q 127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises one or more modified amino acids selected from Group 1C amino acid residues: P27, Y55, K57, T61, E81, K101, and H102; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
[0080] In some cases, the Group 1 amino acids are selected from R35, N92, and T96. In some cases, the FGF polypeptide comprises one or more modified amino acids selected from Group 1 amino acid residues: R35, N92, and T96; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q 127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a Group 1 modified amino acid at R35 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a Group 1 modified amino acid at R35 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, and G120; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a Group 1 modified amino acid at R35 and one or more modified amino acids selected from Group 2 amino acid residues: Q43, Q77, N80, and E82; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a Group 1 modified amino acid at N92 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a Group 1 modified amino acid at N92 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, and G120; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a Group 1 modified amino acid at N92 and a Group 2 modified amino acid at Q43; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a Group 1 modified amino acid at T96 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a Group 1 modified amino acid at T96 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, and G120; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a Group 1 modified amino acid at T96 and a Group 2 modified amino acid at Q43; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
[0081] In some embodiments, the FGF polypeptide is an FGFl polypeptide. In some instances, the FGFl polypeptide comprises one or more modified amino acids selected from Group 1 amino acid residues: P27, R35, D36, R37, Y55, K57, T61, E81, E87, R88, N92, T96, K101, H102, and L133; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the Group 1 modified amino acids are further divided into Group 1A amino acids: R35, D36, and R37; Group IB amino acids: E87, R88, N92, T96, and L133; and Group IC amino acids: P27, Y55, K57, T61, E81, K101, and HI 02. In some instances, the FGFl polypeptide comprises one or more modified amino acids selected from Group 1A amino acid residues: R35, D36, and R37; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGFl polypeptide comprises one or more modified amino acids selected from Group IB amino acid residues: E87, R88, N92, T96, and L133; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGFl polypeptide comprises one or more modified amino acids selected from Group 1C amino acid residues: P27, Y55, K57, T61, E81, K101, and H102; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C- terminal truncation, or a combination thereof. [0082] In some cases, the Group 1 amino acids is selected from R35, N92, and T96. In some cases, the FGFl polypeptide comprises one or more modified amino acids selected from Group 1 amino acid residues: R35, N92, and T96; and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q 127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a Group 1 modified amino acid at R35 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a Group 1 modified amino acid at R35 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, and G120; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a Group 1 modified amino acid at R35 and one or more modified amino acids selected from Group 2 amino acid residues: Q43, Q77, N80, and E82; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a Group 1 modified amino acid at N92 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a Group 1 modified amino acid at N92 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, and G120; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a Group 1 modified amino acid at N92 and a Group 2 modified amino acid at Q43; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a Group 1 modified amino acid at T96 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q 127, and K128; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a Group 1 modified amino acid at T96 and one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, and G120; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a Group 1 modified amino acid at T96 and a Group 2 modified amino acid at Q43; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
[0083] In some embodiments, described herein is a FGF polypeptide comprising a first modification at an amino acid position selected from: 15, 35, 37, 57, 88, 92, 96, 122, and 133; and a second modification at an amino acid position selected from: 1, 2, 5, 6, 7, 9, 10, 17, 35, 37, 38, 40, 43, 45, 47, 48, 49, 50, 51, 53, 57, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 91, 92, 93, 96, 98, 101, 104, 112, 116, 119, 124, 125, 136, 137, 138, 139, and 140; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some instances, the second modification is at an amino acid position selected from: 7, 38, 43, 45, 48, 58, 78, 79, 80, 81, 82, 91, 93, 98, 101, 136, and 137. In some instances, the first modification is selected from an amino acid position: 15, 35, 37, 57, 88, 92, 96, and 122. In some cases, the FGF polypeptide comprises a first modification selected from an amino acid position selected from: 15, 35, 37, 57, 88, 92, 96, and 122; and a second modification at an amino acid position selected from: 1, 2, 5, 6, 7, 9, 10, 17, 35, 37, 38, 40, 43, 45, 47, 48, 49, 50, 51, 53, 57, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 91, 92, 93, 96, 98, 101, 104, 112, 116, 119, 124, 125, 136, 137, 138, 139, and 140; in which the position of the amino acids corresponds to SEQ ID NO: 2.
[0084] In some instances, the first modification is at amino acid position 15. In some cases, the FGF polypeptide comprises a first modification at amino acid position 15, and a second modification at an amino acid position selected from: 1, 2, 5, 6, 7, 9, 10, 17, 37, 38, 40, 43, 45, 47, 48, 49, 50, 51, 53, 57, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 91, 92, 93, 96, 98, 101, 104, 112, 116, 119, 124, 125, 136, 137, 138, 139, and 140; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 15 and a second modification at amino acid position 43, in which the position of the amino acids corresponds to SEQ ID NO: 2.
[0085] In some instances, the first modification is at amino acid position 35. In some cases, the FGF polypeptide comprises a first modification at amino acid position 35, and a second modification at an amino acid position selected from: 1, 2, 5, 6, 7, 9, 10, 17, 37, 38, 40, 43, 45, 47, 48, 49, 50, 51, 53, 57, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 91, 92, 93, 96, 98, 101, 104, 112, 116, 119, 124, 125, 136, 137, 138, 139, 140; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 35, and a second modification at an amino acid position selected from: 1, 2, 5, 6, 9, 10, 17, 37, 43, 45, 47, 48, 49, 50, 51, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 91, 92, 93, 98, 101, 104, 112, 116, 119, 124, 125, 136, 137, 138, and 139; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 35, and a second modification at an amino acid position selected from: 1, 2, 5, 10, 37, 45, 47, 49, 50, 51, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 104, 116, 125, 136, and 139; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 35, and a second modification at an amino acid position selected from: 48, 91, 92, 93, 98, 101, and 137; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 35, and a second modification at an amino acid position: 6, 9, and 17; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 35, and a second modification at an amino acid position: 7, 38, 40, and 140; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 35, and a second modification at an amino acid position 57; in which the position of the amino acids corresponds to SEQ ID NO: 2.
[0086] In some instances, the first modification is at amino acid position 37. In some cases, the FGF polypeptide comprises a first modification at amino acid position 37, and a second modification at an amino acid position selected from: 1, 2, 5, 6, 7, 9, 10, 17, 38, 40, 43, 45, 47, 48, 49, 50, 51, 53, 57, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 91, 92, 93, 96, 98, 101, 104, 112, 116, 119, 124, 125, 136, 137, 138, 139, and 140; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 37, and a second modification at an amino acid position selected from: 43, 53, and 96; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 37, and a second modification at an amino acid position: 43 and 53; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 37, and a second modification at an amino acid position: 92 and 96; in which the position of the amino acids corresponds to SEQ ID NO: 2.
[0087] In some instances, the first modification is at amino acid position 57. In some cases, the FGF polypeptide comprises a first modification at amino acid position 57, and a second modification at an amino acid position selected from: 1, 2, 5, 6, 7, 9, 10, 17, 35, 37, 38, 40, 43, 45, 47, 48, 49, 50, 51, 53, 57, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 91, 92, 93, 96, 98, 101, 104, 112, 116, 119, 124, 125, 136, 137, 138, 139, and 140; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 57, and a second modification at an amino acid position selected from: 10, 48, 90, and 91; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 57, and a second modification at an amino acid position: 10, 48, and 90; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 57, and a second modification at an amino acid position 37; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 57, and a second modification at an amino acid position 92; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 57, and a second modification at an amino acid position 96; in which the position of the amino acids corresponds to SEQ ID NO: 2. [0088] In some instances, the first modification is at amino acid position 88. In some cases, the FGF polypeptide comprises a first modification at amino acid position 88, and a second modification at an amino acid position selected from: 1, 2, 5, 6, 7, 9, 10, 17, 35, 37, 38, 40, 43, 45, 47, 48, 49, 50, 51, 53, 57, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 91, 92, 93, 96, 98, 101, 104, 112, 116, 119, 124, 125, 136, 137, 138, 139, and 140; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 88, and a second modification at an amino acid position: 91 and 96; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 88, and a second modification at an amino acid position 92; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 88, and a second modification at an amino acid position 57; in which the position of the amino acids corresponds to SEQ ID NO: 2.
[0089] In some instances, the first modification is at amino acid position 92. In some cases, the FGF polypeptide comprises a first modification at amino acid position 92, and a second modification at an amino acid position selected from: 1, 2, 5, 6, 7, 9, 10, 17, 35, 37, 38, 40, 43, 45, 47, 48, 49, 50, 51, 53, 57, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 91, 92, 93, 96, 98, 101, 104, 112, 116, 119, 124, 125, 136, 137, 138, 139, and 140; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 92, and a second modification at an amino acid position: 37 and 43; in which the position of the amino acids corresponds to SEQ ID NO: 2.
[0090] In some instances, the first modification is at amino acid position 96. In some cases, the FGF polypeptide comprises a first modification at amino acid position 96, and a second modification at an amino acid position selected from: 1, 2, 5, 6, 7, 9, 10, 17, 35, 37, 38, 40, 43, 45, 47, 48, 49, 50, 51, 53, 57, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 91, 92, 93, 96, 98, 101, 104, 112, 116, 119, 124, 125, 136, 137, 138, 139, and 140; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 96 and a second modification at amino acid position 43, in which the position of the amino acids corresponds to SEQ ID NO: 2.
[0091] In some instances, the first modification is at amino acid position 122. In some cases, the FGF polypeptide comprises a first modification at amino acid position 122, and a second modification at an amino acid position selected from: 1, 2, 5, 6, 7, 9, 10, 17, 35, 37, 38, 40, 43, 45, 47, 48, 49, 50, 51, 53, 57, 58, 59, 70, 72, 76, 77, 78, 79, 80, 81, 82, 90, 91, 92, 93, 96, 98, 101, 104, 112, 116, 119, 124, 125, 136, 137, 138, 139, and 140; in which the position of the amino acids corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at amino acid position 122 and a second modification at amino acid position 35, 37, and 57, in which the position of the amino acids corresponds to SEQ ID NO: 2. [0092] In some embodiments, the FGF polypeptide comprises a first modification at an amino acid residue selected from selected from: Y15, R35, R37, K57, R88, N92, T96, R122, and L133; and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S 116, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the second modification is at an amino acid residue selected from: N7, S38, Q43, Q45, A48, S58, T78, P79, N80, E81, E82, E91, H93, 198, K101, P136, and V137. In some instances, the first modification is selected from: Y15, R35, R37, K57, R88, N92, T96, and R122. In some instances, the FGF polypeptide comprises a first modification at an amino acid residue selected from selected from: Y15, R35, R37, K57, R88, N92, T96, and R122; and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, SI 16, Rl 19, H124, Y125, P136, V137, S138, S139, and D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, Y15, R35, R37, K57, R88, N92, T96, and R122 are independently mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, K112, SI 16, R119, H124, Y125, P136, V137, S 138, S 139, D140 are independently mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, the FGF polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N- terminal truncation comprises a deletion of amino acid residues 1-9.
[0093] In some instances, the first modification is Y 15. In some cases, the FGF polypeptide comprises a first modification at Y15, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at Y15, and a second modification at Q43, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, Y 15 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, Y15 is mutated to phenylalanine (F). In some cases, the FGF polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C- terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0094] In some instances, the first modification is R35. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S 138, S 139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, K9, K10, S 17, R37, Q43, Q45, S47, A48, E49, S50, V51, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, 198, K101, E104, Kl 12, SI 16, Rl 19, H124, Y125, P136, V137, S138, and S 139; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: Fl, N2, P5, K10, R37, Q45, S47, E49, S50, V51, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E104, SI 16, Y125, P136, and S139; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: A48, E91, N92, H93, 198, K101, and V137; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: Kl 12, Rl 19, and H124; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from selected from: G6, K9, and S17; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at K57; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at Kl 12; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at Rl 19; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at HI 24; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at N7; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at S38; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at Q40; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at D 140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at E81; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R35, and a second modification at R37; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, R35 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, R35 is mutated to alanine (A) or glutamic acid (E). In some cases, the FGF polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0095] In some instances, the first modification is R37. In some cases, the FGF polypeptide comprises a first modification at R37, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S 138, S 139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R37, and a second modification at an amino acid residue selected from: Q43, E53, and T96; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R37, and a second modification at an amino acid residue selected from: Q43 and E53; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R37, and a second modification at an amino acid residue selected from: N92 and T96; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, R37 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, R37 is mutated to alanine (A) or glutamic acid (E). In some cases, the FGF polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0096] In some instances, the first modification is K57. In some cases, the FGF polypeptide comprises a first modification at K57, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S 138, S 139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at K57, and a second modification at an amino acid residue selected from: K10, A48, E90, and E91; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at K57, and a second modification at an amino acid residue selected from: K10, A48, and E90; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at K57, and a second modification at R37; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at K57, and a second modification at N92; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at K57, and a second modification at T96; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, K57 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, K57 is mutated to alanine (A), asparagine (N), glutamic acid (E), or histidine (H). In some cases, the FGF polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0097] In some instances, the first modification is R88. In some cases, the FGF polypeptide comprises a first modification at R88, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R88, and a second modification at an amino acid residue selected from: E91 and T96; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R88, and a second modification at N92; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R88, and a second modification at K57; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, R88 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, R88 is mutated to leucine (L). In some cases, the FGF polypeptide comprises a truncation. In some cases, the truncation is an N- terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0098] In some instances, the first modification is N92. In some cases, the FGF polypeptide comprises a first modification at N92, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at N92, and a second modification at an amino acid residue selected from: R37 and Q43; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, N92 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, N92 is mutated to alanine (A) or threonine (T). In some cases, the FGF polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0099] In some instances, the first modification is T96. In some cases, the FGF polypeptide comprises a first modification at T96, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at T96, and a second modification at Q43, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, T96 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, T96 is mutated to alanine (A). In some cases, the FGF polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0100] In some instances, the first modification is R122. In some cases, the FGF polypeptide comprises a first modification at R122, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R122, and a second modification at an amino acid residue selected from: R35, R37, and K57, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R122, and a second modification at R35, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R122, and a second modification at R37, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide comprises a first modification at R122, and a second modification at K57, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, R122 is mutated to an amino acid selected from:
alanine (A), aspartic acid (D), glutamic acid (E), phenylalanine (F), glycine (G), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), glutamine (Q), serine (S), valine (V), and tyrosine (Y). In some cases, R122 is mutated to an amino acid selected from: : aspartic acid (D), phenylalanine (F), glycine (G), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), glutamine (Q), serine (S), valine (V), and tyrosine (Y). In some cases, R122 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, R122 is mutated to glutamic acid (E). In some cases, R122 is mutated to alanine (A). In some cases, R122 is mutated to aspartic acid (D). In some cases, R122 is mutated to phenylalanine (F). In some cases, R122 is mutated to glycine (G). In some cases, R122 is mutated to histidine (H). In some cases, R122 is mutated to isoleucine (I). In some cases, R122 is mutated to lysine (K). In some cases, R122 is mutated to leucine (L). In some cases, R122 is mutated to methionine (M). In some cases, R122 is mutated to asparagine (N). In some cases, R122 is mutated to glutamine (Q). In some cases, R122 is mutated to serine (S). In some cases, R122 is mutated to valine (V). In some cases, R122 is mutated to tyrosine (Y). In some cases, the FGF polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C- terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0101] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some embodiments, the FGF1 polypeptide comprises a first modification at an amino acid residue selected from: Y15, R35, R37, K57, R88, N92, T96, R122, and L133; and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S 17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, SI 16, Rl 19, H124, Y125, P136, V137, S 138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the second modification is at an amino acid residue selected from: N7, S38, Q43, Q45, A48, S58, T78, P79, N80, E81, E82, E91, H93, 198, K101, P136, and V137. In some instances, the first modification is selected from: Y15, R35, R37, K57, R88, N92, T96, and R122. In some instances, the FGF1 polypeptide comprises a first modification at an amino acid residue selected from: Y15, R35, R37, K57, R88, N92, T96, and R122; and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, SI 16, Rl 19, HI 24, Y125, PI 36, VI 37, S138, S 139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, Y15, R35, R37, K57, R88, N92, T96, and R122 are independently mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, Fl, N2, P5, G6, N7, K9, K10, S 17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, SI 16, Rl 19, H124, Y125, P136, V137, S 138, S139, D140are independently mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, the FGFl polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0102] In some instances, the first modification is Y15. In some cases, the FGFl polypeptide comprises a first modification at Y15, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at Y15, and a second modification at Q43, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, Y 15 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, Y15 is mutated to phenylalanine (F). In some cases, the FGFl polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C- terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0103] In some instances, the first modification is R35. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S 138, S 139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, K9, K10, S17, R37, Q43, Q45, S47, A48, E49, S50, V51, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, and S139; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: Fl, N2, P5, K10, R37, Q45, S47, E49, S50, V51, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E104, SI 16, Y125, P136, and S 139; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: A48, E91, N92, H93, 198, K101, and V137; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: G6, K9, and S I 7; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at an amino acid residue selected from: Kl 12, Rl 19, and H124; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at K57; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at Kl 12; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at Rl 19; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at HI 24; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at N7; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at S38; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at Q40; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at D 140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at E81; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R35, and a second modification at R37; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, R35 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, R35 is mutated to alanine (A) or glutamic acid (E). In some cases, the FGFl polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0104] In some instances, the first modification is R37. In some cases, the FGFl polypeptide comprises a first modification at R37, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S 138, S 139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R37, and a second modification at an amino acid residue selected from: Q43, E53, and T96; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF1 polypeptide comprises a first modification at R37, and a second modification at an amino acid residue selected from: Q43 and E53; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF1 polypeptide comprises a first modification at R37, and a second modification at an amino acid residue selected from: N92 and T96; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, R37 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, R37 is mutated to alanine (A) or glutamic acid (E). In some cases, the FGF1 polypeptide comprises a truncation. In some cases, the truncation is an N- terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0105] In some instances, the first modification is K57. In some cases, the FGF1 polypeptide comprises a first modification at K57, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S 138, S 139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF1 polypeptide comprises a first modification at K57, and a second modification at an amino acid residue selected from: K10, A48, E90, and E91; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF1 polypeptide comprises a first modification at K57, and a second modification at an amino acid residue selected from: K10, A48, and E90; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF1 polypeptide comprises a first modification at K57, and a second modification at R37; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF1 polypeptide comprises a first modification at K57, and a second modification at N92; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF1 polypeptide comprises a first modification at K57, and a second modification at T96; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, K57 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, K57 is mutated to alanine (A), asparagine (N), glutamic acid (E), or histidine (H). In some cases, the FGF1 polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9. [0106] In some instances, the first modification is R88. In some cases, the FGF1 polypeptide comprises a first modification at R88, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF1 polypeptide comprises a first modification at R88, and a second modification at an amino acid residue selected from: E91 and T96; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF1 polypeptide comprises a first modification at R88, and a second modification at N92; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF1 polypeptide comprises a first modification at R88, and a second modification at K57; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, R88 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, R88 is mutated to leucine (L). In some cases, the FGF1 polypeptide comprises a truncation. In some cases, the truncation is an N- terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0107] In some instances, the first modification is N92. In some cases, the FGF1 polypeptide comprises a first modification at N92, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF1 polypeptide comprises a first modification at N92, and a second modification at an amino acid residue selected from: R37 and Q43; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, N92 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, N92 is mutated to alanine (A) or threonine (T). In some cases, the FGF1 polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0108] In some instances, the first modification is T96. In some cases, the FGF1 polypeptide comprises a first modification at T96, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at T96, and a second modification at Q43, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, T96 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, T96 is mutated to alanine (A). In some cases, the FGFl polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0109] In some instances, the first modification is R122. In some cases, the FGFl polypeptide comprises a first modification at R122, and a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, Kl 12, S I 16, Rl 19, H124, Y125, P136, V137, S138, S139, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R122, and a second modification at an amino acid residue selected from: R35, R37, and K57, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R122, and a second modification at R35, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R122, and a second modification at R37, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGFl polypeptide comprises a first modification at R122, and a second modification at K57, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, R122 is mutated to an amino acid selected from:
alanine (A), aspartic acid (D), glutamic acid (E), phenylalanine (F), glycine (G), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), glutamine (Q), serine (S), valine (V), and tyrosine (Y). In some cases, R122 is mutated to an amino acid selected from: : aspartic acid (D), phenylalanine (F), glycine (G), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), glutamine (Q), serine (S), valine (V), and tyrosine (Y). In some cases, R122 is mutated to an amino acid selected from: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F). In some cases, R122 is mutated to glutamic acid (E). In some cases, R122 is mutated to alanine (A). In some cases, R122 is mutated to aspartic acid (D). In some cases, R122 is mutated to phenylalanine (F). In some cases, R122 is mutated to glycine (G). In some cases, R122 is mutated to histidine (H). In some cases, R122 is mutated to isoleucine (I). In some cases, R122 is mutated to lysine (K). In some cases, R122 is mutated to leucine (L). In some cases, R122 is mutated to methionine (M). In some cases, R122 is mutated to asparagine (N). In some cases, R122 is mutated to glutamine (Q). In some cases, R122 is mutated to serine (S). In some cases, R122 is mutated to valine (V). In some cases, R122 is mutated to tyrosine (Y). In some cases, the FGF1 polypeptide comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C- terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0110] In some embodiments, described herein is a FGF polypeptide comprising a modification at amino acid position 48, 53, 57, 58, 72, 76, 77, 78, 79, 96, or 98, wherein the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the modification is at amino acid position 57, 77, or 96. In some cases, the FGF polypeptide comprises a modification at amino acid position 48. In some cases, the FGF polypeptide comprises a modification at amino acid position 53. In some cases, the FGF polypeptide comprises a modification at amino acid position 57. In some cases, the FGF polypeptide comprises a modification at amino acid position 58. In some cases, the FGF polypeptide comprises a modification at amino acid position 72. In some cases, the FGF polypeptide comprises a modification at amino acid position 76. In some cases, the FGF polypeptide comprises a modification at amino acid position 77. In some cases, the FGF polypeptide comprises a modification at amino acid position 78. In some cases, the FGF polypeptide comprises a modification at amino acid position 79. In some cases, the FGF polypeptide comprises a modification at amino acid position 96. In some cases, the FGF polypeptide comprises a modification at amino acid position 98. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0111] In some embodiments, the FGF polypeptide comprises a modification at amino acid residue A48, E53, K57, S58, L72, S76, Q77, T78, P79, T96, or 198, wherein the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the modification is K57, Q77, or T97. In some instances, the FGF polypeptide comprises a modification at A48. In some instances, the FGF polypeptide comprises a modification at E53. In some instances, the FGF polypeptide comprises a modification at K57. In some instances, the FGF polypeptide comprises a modification at S58. In some instances, the FGF polypeptide comprises a modification at L72. In some instances, the FGF polypeptide comprises a modification at S76. In some instances, the FGF polypeptide comprises a modification at Q77. In some instances, the FGF polypeptide comprises a modification at T78. In some instances, the FGF polypeptide comprises a modification at P79. In some instances, the FGF polypeptide comprises a modification at T96. In some instances, the FGF polypeptide comprises a modification at 198. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0112] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some embodiments, the FGF1 polypeptide comprises a modification at amino acid residue A48, E53, K57, S58, L72, S76, Q77, T78, P79, T96, or 198, wherein the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the modification is K57, Q77, or T97. In some instances, the FGF1 polypeptide comprises a modification at A48. In some instances, the FGF1 polypeptide comprises a modification at E53. In some instances, the FGF1 polypeptide comprises a modification at K57. In some instances, the FGF1 polypeptide comprises a modification at S58. In some instances, the FGF1 polypeptide comprises a modification at L72. In some instances, the FGF1 polypeptide comprises a modification at S76. In some instances, the FGF1 polypeptide comprises a modification at Q77. In some instances, the FGF1 polypeptide comprises a modification at T78. In some instances, the FGF1 polypeptide comprises a modification at P79. In some instances, the FGF1 polypeptide comprises a modification at T96. In some instances, the FGF1 polypeptide comprises a modification at 198. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0113] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 35 and a second modification at an amino acid position selected from: 3, 4, 7, 8, 11, 15, 17, 18, 21, 26, 27, 28, 30, 32, 34, 36, 38, 39, 40, 41, 43, 53, 55, 57, 60, 61, 63, 68, 69, 82, 84, 86, 87, 88, 89, 90, 92, 94, 95, 96, 100, 101, 102, 105, 106, 108, 112, 113, 118, 119, 121, 127, 128, 131, 133, 134, 135, and 140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N- terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0114] In some embodiments, described herein is a FGF polypeptide comprising a first modification at R35 and a second modification at an amino acid residue selected from: L3, P4, N7, Y8, Pl l, Y15, S17, N18, H21, L26, P27, D28, T30, D32, T34, D36, S38, D39, Q40, H41, Q43, E53, Y55, K57, E60, T61, Q63, D68, T69, E82, L84, L86, E87, R88, L89, E90, N92, Y94, N95, T96, K100, K101, H102, K105, N106, F108, K112, K113, K118, R119, P121, Q127, K128, L131, L133, P134, L135, and D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at L3. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at P4. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at N7. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Y8. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at PI 1. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Y 15. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at S17. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at N18. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at H21. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at L26. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at P27. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at T30. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at D32. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at T34. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at S38. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at D39. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Q40. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at H41. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Q43. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at E53. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Y55. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at K57. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at E60. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at T61. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Q63. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at D68. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at T69. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at E82. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at L86. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at L89. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at E90. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at N92. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at T96. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at K 100. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Kl 01. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at HI 02. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at K105. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at N106. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at F108. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Kl 12. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Kl 13. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Kl 18. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Rl 19. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at P 121. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at Q127. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at K128. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at L131. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at LI 33. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at P134. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at L135. In some instances, the FGF polypeptide comprises a first modification at R35 and a second modification at D 140. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9. [0115] In some embodiments, the FGF polypeptide is an FGFl polypeptide. In some instances, the FGFl polypeptide comprising a first modification at R35 and a second modification at an amino acid residue selected from: L3, P4, N7, Y8, Pl l, Y15, S17, N18, H21, L26, P27, D28, T30, D32, T34, D36, S38, D39, Q40, H41, Q43, E53, Y55, K57, E60, T61, Q63, D68, T69, E82, L84, L86, E87, R88, L89, E90, N92, Y94, N95, T96, K100, K101, H102, K105, N106, F108, Kl 12, Kl 13, Kl 18, Rl 19, P121, Q127, K128, L131, L133, P134, L135, D140; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at L3. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at P4. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at N7. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Y8. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at PI 1. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Y15. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at S 17. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at N 18. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at H21. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at L26. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at P27. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at D28. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at T30. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at D32. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at T34. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at D36. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at S38. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at D39. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Q40. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at H41. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Q43. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at E53. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Y55. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at K57. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at E60. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at T61. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Q63. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at D68. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at T69. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at E82. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at L84. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at L86. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at E87. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at R88. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at L89. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at E90. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at N92. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Y94. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at N95. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at T96. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at K 100. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Kl 01. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at H102. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at K 105. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at N 106. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at F 108. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Kl 12. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Kl 13. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Kl 18. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Rl 19. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at P 121. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at Q127. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at K128. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at L131. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at LI 33. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at P134. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at L 135. In some instances, the FGFl polypeptide comprises a first modification at R35 and a second modification at D 140. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N- terminal truncation comprises a deletion of amino acid residues 1-9.
[0116] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 88 and a second modification at an amino acid position selected from: 43, 84, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0117] In some embodiments, described herein is a FGF polypeptide comprising a first modification at R88 and a second modification at an amino acid residue selected from: Q43, L84, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at R88 and a second modification at Q43. In some instances, the FGF polypeptide comprises a first modification at R88 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at R88 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non- polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N- terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0118] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some instances, the FGF1 polypeptide comprising a first modification at R88 and a second modification at an amino acid residue selected from: Q43, L84, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF1 polypeptide comprises a first
modification at R88 and a second modification at Q43. In some instances, the FGF1 polypeptide comprises a first modification at R88 and a second modification at L84. In some instances, the FGF1 polypeptide comprises a first modification at R88 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N- terminal truncation comprises a deletion of amino acid residues 1-9.
[0119] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 92 and a second modification at amino acid position 96, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprising a first modification at N92 and a second modification at T96, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide is an FGF1 polypeptide. In some cases, the FGF1 polypeptide comprises a first modification at N92 and a second modification at T96, in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0120] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 118 and a second modification at an amino acid position selected from: Q43 and L84; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some embodiments, the FGF polypeptide comprising a first modification at Kl 18 and a second modification at an amino acid residue selected from: Q43 and L84; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at Kl 18 and a second modification at Q43. In some instances, the FGF polypeptide comprises a first modification at Kl 18 and a second modification at L84. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N- terminal truncation comprises a deletion of amino acid residues 1-9.
[0121] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some cases, the FGF1 polypeptide comprises a first modification at Kl 18 and a second modification at an amino acid residue selected from: Q43 and L84; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF1 polypeptide comprises a first modification at Kl 18 and a second modification at Q43. In some instances, the FGF 1 polypeptide comprises a first modification at Kl 18 and a second modification at L84. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0122] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 133 and a second modification at an amino acid position selected from: 34, 37, 43, 84, 88, 95, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0123] In some embodiments, described herein is a FGF polypeptide comprising a first modification at L133 and a second modification at an amino acid residue selected from: T34, R37, Q43, L84, R88, N95, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at L133 and a second modification at T34. In some instances, the FGF polypeptide comprises a first modification at L133 and a second modification at R37. In some instances, the FGF polypeptide comprises a first modification at LI 33 and a second modification at Q43. In some instances, the FGF polypeptide comprises a first modification at L133 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at LI 33 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at LI 33 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at L133 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0124] In some embodiments, the FGF polypeptide is an FGFl polypeptide. In some instances, the FGFl polypeptide comprising a first modification at LI 33 and a second modification at an amino acid residue selected from: T34, R37, Q43, L84, R88, N95, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGFl polypeptide comprises a first modification at L133 and a second modification at T34. In some instances, the FGFl polypeptide comprises a first modification at LI 33 and a second modification at R37. In some instances, the FGFl polypeptide comprises a first modification at LI 33 and a second modification at Q43. In some instances, the FGFl polypeptide comprises a first modification at L133 and a second modification at L84. In some instances, the FGFl polypeptide comprises a first modification at L133 and a second modification at R88. In some instances, the FGFl polypeptide comprises a first modification at L133 and a second modification at N95. In some instances, the FGFl polypeptide comprises a first modification at LI 33 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGFl polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N- terminal truncation comprises a deletion of amino acid residues 1-9.
[0125] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 10 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0126] In some embodiments, described herein is a FGF polypeptide comprising a first modification at K10 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at Y 15. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at SI 16. In some instances, the FGF polypeptide comprises a first modification at K10 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non- polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N- terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0127] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some instances, the FGF1 polypeptide comprising a first modification at K10 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, SI 16, and K118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at Y 15. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at D28. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at R35. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at D36. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at L84. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at E87. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at R88. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at Y94. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at N95. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at K101. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at SI 16. In some instances, the FGF1 polypeptide comprises a first modification at K10 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N- terminal truncation comprises a deletion of amino acid residues 1-9.
[0128] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 116 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9. [0129] In some embodiments, described herein is a FGF polypeptide comprising a first modification at S I 16 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at S I 16 and a second modification at Y 15. In some instances, the FGF polypeptide comprises a first modification at S I 16 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at S I 16 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at SI 16 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at S I 16 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at S I 16 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at S I 16 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at S 116 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at S 116 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at SI 16 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at SI 16 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0130] In some embodiments, the FGF polypeptide is an FGFl polypeptide. In some instances, the FGFl polypeptide comprises a first modification at SI 16 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGFl polypeptide comprises a first modification at S 116 and a second modification at Y 15. In some instances, the FGFl polypeptide comprises a first modification at SI 16 and a second modification at D28. In some instances, the FGFl polypeptide comprises a first modification at SI 16 and a second modification at R35. In some instances, the FGFl polypeptide comprises a first modification at SI 16 and a second modification at D36. In some instances, the FGFl polypeptide comprises a first modification at S I 16 and a second modification at L84. In some instances, the FGFl polypeptide comprises a first modification at S I 16 and a second modification at E87. In some instances, the FGFl polypeptide comprises a first modification at S I 16 and a second modification at R88. In some instances, the FGFl polypeptide comprises a first modification at SI 16 and a second modification at Y94. In some instances, the FGFl polypeptide comprises a first modification at SI 16 and a second modification at N95. In some instances, the FGF 1 polypeptide comprises a first modification at S 116 and a second modification at K101. In some instances, the FGF1 polypeptide comprises a first modification at SI 16 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0131] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 122 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0132] In some embodiments, described herein is a FGF polypeptide comprising a first modification at R122 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at Y15. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at R122 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0133] In some embodiments, the FGF polypeptide is an FGFl polypeptide. In some instances, the FGFl polypeptide comprising a first modification at R122 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, SI 16, and K118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at Y 15. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at D28. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at R35. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at D36. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at L84. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at E87. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at R88. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at Y94. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at N95. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at K101. In some instances, the FGFl polypeptide comprises a first modification at R122 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGFl polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0134] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 124 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0135] In some embodiments, described herein is a FGF polypeptide comprising a first modification at H124 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at Y15. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at HI 24 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at HI 24 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at H124 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at HI 24 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0136] In some embodiments, the FGF polypeptide is an FGFl polypeptide. In some instances, the FGFl polypeptide comprising a first modification at HI 24 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, SI 16, and K118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGFl polypeptide comprises a first modification at H124 and a second modification at Y15. In some instances, the FGFl polypeptide comprises a first modification at HI 24 and a second modification at D28. In some instances, the FGFl polypeptide comprises a first modification at H124 and a second modification at R35. In some instances, the FGFl polypeptide comprises a first modification at HI 24 and a second modification at D36. In some instances, the FGFl polypeptide comprises a first modification at H124 and a second modification at L84. In some instances, the FGFl polypeptide comprises a first modification at H124 and a second modification at E87. In some instances, the FGFl polypeptide comprises a first modification at HI 24 and a second modification at R88. In some instances, the FGFl polypeptide comprises a first modification at HI 24 and a second modification at Y94. In some instances, the FGFl polypeptide comprises a first modification at H124 and a second modification at N95. In some instances, the FGFl polypeptide comprises a first modification at H124 and a second modification at K101. In some instances, the FGFl polypeptide comprises a first modification at H124 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0137] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 64 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0138] In some embodiments, described herein is a FGF polypeptide comprising a first modification at Y64 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at Y 15. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at Y64 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9. [0139] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some instances, the FGF1 polypeptide comprising a first modification at Y64 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, SI 16, and K118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF1 polypeptide comprises a first modification at Y64 and a second modification at Y 15. In some instances, the FGF 1 polypeptide comprises a first modification at Y64 and a second modification at D28. In some instances, the FGF1 polypeptide comprises a first modification at Y64 and a second modification at R35. In some instances, the FGF1 polypeptide comprises a first modification at Y64 and a second modification at D36. In some instances, the FGF1 polypeptide comprises a first modification at Y64 and a second modification at L84. In some instances, the FGF1 polypeptide comprises a first modification at Y64 and a second modification at E87. In some instances, the FGF1 polypeptide comprises a first modification at Y64 and a second modification at R88. In some instances, the FGF 1 polypeptide comprises a first modification at Y64 and a second modification at Y94. In some instances, the FGF1 polypeptide comprises a first modification at Y64 and a second modification at N95. In some instances, the FGF1 polypeptide comprises a first modification at Y64 and a second modification at K101. In some instances, the FGF1 polypeptide comprises a first modification at Y64 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0140] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 60 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0141] In some embodiments, described herein is a FGF polypeptide comprising a first modification at E60 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at E60 and a second modification at Y 15. In some instances, the FGF polypeptide comprises a first modification at E60 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at E60 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at E60 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at E60 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at E60 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at E60 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at E60 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at E60 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at E60 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at E60 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0142] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some instances, the FGF1 polypeptide comprising a first modification at E60 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, SI 16, and K118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF1 polypeptide comprises a first modification at E60 and a second modification at Y 15. In some instances, the FGF 1 polypeptide comprises a first modification at E60 and a second modification at D28. In some instances, the FGF1 polypeptide comprises a first modification at E60 and a second modification at R35. In some instances, the FGF1 polypeptide comprises a first modification at E60 and a second modification at D36. In some instances, the FGF1 polypeptide comprises a first modification at E60 and a second modification at L84. In some instances, the FGF1 polypeptide comprises a first modification at E60 and a second modification at E87. In some instances, the FGF1 polypeptide comprises a first modification at E60 and a second modification at R88. In some instances, the FGF 1 polypeptide comprises a first modification at E60 and a second modification at Y94. In some instances, the FGF1 polypeptide comprises a first modification at E60 and a second modification at N95. In some instances, the FGF1 polypeptide comprises a first modification at E60 and a second modification at K101. In some instances, the FGF1 polypeptide comprises a first modification at E60 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0143] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 53 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0144] In some embodiments, described herein is a FGF polypeptide comprising a first modification at E53 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at E53 and a second modification at Y 15. In some instances, the FGF polypeptide comprises a first modification at E53 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at E53 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at E53 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at E53 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at E53 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at E53 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at E53 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at E53 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at E53 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at E53 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0145] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some instances, the FGF1 polypeptide comprising a first modification at E53 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, SI 16, and K118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF1 polypeptide comprises a first modification at E53 and a second modification at Y 15. In some instances, the FGF1 polypeptide comprises a first modification at E53 and a second modification at D28. In some instances, the FGF1 polypeptide comprises a first modification at E53 and a second modification at R35. In some instances, the FGF1 polypeptide comprises a first modification at E53 and a second modification at D36. In some instances, the FGF1 polypeptide comprises a first modification at E53 and a second modification at L84. In some instances, the FGF1 polypeptide comprises a first modification at E53 and a second modification at E87. In some instances, the FGF1 polypeptide comprises a first modification at E53 and a second modification at R88. In some instances, the FGF1 polypeptide comprises a first modification at E53 and a second modification at Y94. In some instances, the FGF1 polypeptide comprises a first modification at E53 and a second modification at N95. In some instances, the FGF1 polypeptide comprises a first modification at E53 and a second modification at K101. In some instances, the FGF1 polypeptide comprises a first modification at E53 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0146] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 49 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0147] In some embodiments, described herein is a FGF polypeptide comprising a first modification at E49 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at E49 and a second modification at Y 15. In some instances, the FGF polypeptide comprises a first modification at E49 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at E49 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at E49 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at E49 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at E49 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at E49 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at E49 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at E49 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at E49 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at E49 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0148] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some instances, the FGF1 polypeptide comprising a first modification at E49 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, SI 16, and K118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF1 polypeptide comprises a first modification at E49 and a second modification at Y 15. In some instances, the FGF 1 polypeptide comprises a first modification at E49 and a second modification at D28. In some instances, the FGF1 polypeptide comprises a first modification at E49 and a second modification at R35. In some instances, the FGF1 polypeptide comprises a first modification at E49 and a second modification at D36. In some instances, the FGF1 polypeptide comprises a first modification at E49 and a second modification at L84. In some instances, the FGF1 polypeptide comprises a first modification at E49 and a second modification at E87. In some instances, the FGF1 polypeptide comprises a first modification at E49 and a second modification at R88. In some instances, the FGF 1 polypeptide comprises a first modification at E49 and a second modification at Y94. In some instances, the FGF1 polypeptide comprises a first modification at E49 and a second modification at N95. In some instances, the FGF1 polypeptide comprises a first modification at E49 and a second modification at K101. In some instances, the FGF1 polypeptide comprises a first modification at E49 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0149] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 47 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0150] In some embodiments, described herein is a FGF polypeptide comprising a first modification at S47 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at S47 and a second modification at Y 15. In some instances, the FGF polypeptide comprises a first modification at S47 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at S47 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at S47 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at S47 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at S47 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at S47 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at S47 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at S47 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at S47 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at S47 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0151] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some instances, the FGF1 polypeptide comprising a first modification at S47 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, SI 16, and K118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at Y 15. In some instances, the FGF 1 polypeptide comprises a first modification at S47 and a second modification at D28. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at R35. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at D36. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at L84. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at E87. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at R88. In some instances, the FGF 1 polypeptide comprises a first modification at S47 and a second modification at Y94. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at N95. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at K101. In some instances, the FGF1 polypeptide comprises a first modification at S47 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0152] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 43 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0153] In some embodiments, described herein is a FGF polypeptide comprising a first modification at Q43 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at Q43 and a second modification at Y 15. In some instances, the FGF polypeptide comprises a first modification at Q43 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at Q43 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at Q43 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at Q43 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at Q43 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at Q43 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at Q43 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at Q43 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at Q43 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at Q43 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0154] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some instances, the FGF1 polypeptide comprising a first modification at Q43 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, SI 16, and K118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF1 polypeptide comprises a first modification at Q43 and a second modification at Y 15. In some instances, the FGF 1 polypeptide comprises a first modification at Q43 and a second modification at D28. In some instances, the FGF1 polypeptide comprises a first modification at Q43 and a second modification at R35. In some instances, the FGF1 polypeptide comprises a first modification at Q43 and a second modification at D36. In some instances, the FGF1 polypeptide comprises a first modification at Q43 and a second modification at L84. In some instances, the FGF1 polypeptide comprises a first modification at Q43 and a second modification at E87. In some instances, the FGF1 polypeptide comprises a first modification at Q43 and a second modification at R88. In some instances, the FGF 1 polypeptide comprises a first modification at Q43 and a second modification at Y94. In some instances, the FGF1 polypeptide comprises a first modification at Q43 and a second modification at N95. In some instances, the FGF1 polypeptide comprises a first modification at Q43 and a second modification at K101. In some instances, the FGF1 polypeptide comprises a first modification at Q43 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0155] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 40 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0156] In some embodiments, described herein is a FGF polypeptide comprising a first modification at Q40 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at Q40 and a second modification at Y 15. In some instances, the FGF polypeptide comprises a first modification at Q40 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at Q40 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at Q40 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at Q40 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at Q40 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at Q40 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at Q40 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at Q40 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at Q40 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at Q40 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N- terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0157] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some instances, the FGF1 polypeptide comprising a first modification at Q40 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, SI 16, and K118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF1 polypeptide comprises a first modification at Q40 and a second modification at Y 15. In some instances, the FGF 1 polypeptide comprises a first modification at Q40 and a second modification at D28. In some instances, the FGF1 polypeptide comprises a first modification at Q40 and a second modification at R35. In some instances, the FGF1 polypeptide comprises a first modification at Q40 and a second modification at D36. In some instances, the FGF1 polypeptide comprises a first modification at Q40 and a second modification at L84. In some instances, the FGF1 polypeptide comprises a first modification at Q40 and a second modification at E87. In some instances, the FGF1 polypeptide comprises a first modification at Q40 and a second modification at R88. In some instances, the FGF 1 polypeptide comprises a first modification at Q40 and a second modification at Y94. In some instances, the FGF1 polypeptide comprises a first modification at Q40 and a second modification at N95. In some instances, the FGF1 polypeptide comprises a first modification at Q40 and a second modification at K101. In some instances, the FGF1 polypeptide comprises a first modification at Q40 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0158] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 17 and a second modification at an amino acid position selected from: 15, 28, 35, 36, 84, 87, 88, 94, 95, 101, 116, and 118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0159] In some embodiments, described herein is a FGF polypeptide comprising a first modification at S 17 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, S I 16, and Kl 18; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprises a first modification at S 17 and a second modification at Y 15. In some instances, the FGF polypeptide comprises a first modification at S17 and a second modification at D28. In some instances, the FGF polypeptide comprises a first modification at S 17 and a second modification at R35. In some instances, the FGF polypeptide comprises a first modification at S 17 and a second modification at D36. In some instances, the FGF polypeptide comprises a first modification at S 17 and a second modification at L84. In some instances, the FGF polypeptide comprises a first modification at S17 and a second modification at E87. In some instances, the FGF polypeptide comprises a first modification at S17 and a second modification at R88. In some instances, the FGF polypeptide comprises a first modification at S 17 and a second modification at Y94. In some instances, the FGF polypeptide comprises a first modification at S17 and a second modification at N95. In some instances, the FGF polypeptide comprises a first modification at S 17 and a second modification at K101. In some instances, the FGF polypeptide comprises a first modification at S 17 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0160] In some embodiments, the FGF polypeptide is an FGF1 polypeptide. In some instances, the FGF1 polypeptide comprising a first modification at S 17 and a second modification at an amino acid residue selected from: Y15, D28, R35, D36, L84, E87, R88, Y94, N95, K101, SI 16, and K118; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF1 polypeptide comprises a first modification at S 17 and a second modification at Y 15. In some instances, the FGF1 polypeptide comprises a first modification at S17 and a second modification at D28. In some instances, the FGF1 polypeptide comprises a first modification at S 17 and a second modification at R35. In some instances, the FGF1 polypeptide comprises a first modification at S17 and a second modification at D36. In some instances, the FGF1 polypeptide comprises a first modification at S 17 and a second modification at L84. In some instances, the FGF1 polypeptide comprises a first modification at S17 and a second modification at E87. In some instances, the FGF1 polypeptide comprises a first modification at S 17 and a second modification at R88. In some instances, the FGF1 polypeptide comprises a first modification at S17 and a second modification at Y94. In some instances, the FGF1 polypeptide comprises a first modification at S 17 and a second modification at N95. In some instances, the FGF1 polypeptide comprises a first modification at S17 and a second modification at K101. In some instances, the FGF1 polypeptide comprises a first modification at S 17 and a second modification at Kl 18. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0161] In some embodiments, described herein is a FGF polypeptide comprising a first modification at amino acid position 15, a second modification at amino acid position 133, and a third modification at amino acid position 92; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some instances, the FGF polypeptide comprising a first modification at Y 15, a second modification at LI 33, and a third modification at N92; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the FGF polypeptide is an FGF1 polypeptide. In some cases, the FGF1 polypeptide comprising a first modification at Y 15, a second modification at LI 33, and a third modification at N92; in which the numbering of the amino acid residues corresponds to SEQ ID NO: 2. In some cases, the modification comprises a mutation to a non-polar residue or a charged residue. In some cases, the modification comprises a mutation to alanine. In some cases, the FGF1 polypeptide further comprises a truncation. In some cases, the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof. In some cases, the truncation is an N-terminal truncation. In some cases, the N-terminal truncation comprises a deletion of amino acid residues 1-9.
[0162] In some embodiments, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Tables 2-12. In some instances, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Table 2. In some instances, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Table 3. In some instances, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Table 4. In some instances, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Table 5. In some instances, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Table 6. In some instances, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Table 7. In some instances, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Table 8. In some instances, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Table 9. In some instances, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Table 10. In some instances, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Table 11. In some instances, an isolated and purified FGF polypeptide described herein comprises an FGFl polypeptide disclosed in Table 12.
[0163] In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 80% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 85% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 90% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 91% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 92% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 93% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 94% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 95% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 96% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 97% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 98% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 99% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide comprising about 100% sequence identity to a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974. In some instances, the FGF mutant is an FGFl polypeptide consists of a polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
[0164] In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 139, 141, 142, 143, 144, 146, 147, 148, 149, 150, 151, 152, 153, 156, 157, 158, 159, 163, 165, 168, 170, 171, 174, 177, 284, 286, 290, 401, 484, 488, or 518. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 130. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 131. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 132. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 133. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 134. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 135. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 139. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 141. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 142. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 143. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 144. In some embodiments, the FGF mutant is an FGFl polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 146. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 147. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 148. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 149. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 150. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 151. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 152. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 153. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 156. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 157. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 158. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 159. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 163. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 165. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 168. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 170. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 171. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 174. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 177. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 284. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 286. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 290. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 401. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 484. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 488. In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a polypeptide of SEQ ID NO: 518.
[0165] In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 100% sequence identity to a polypeptide of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 139, 141, 142, 143, 144, 146, 147, 148, 149, 150, 151, 152, 153, 156, 157, 158, 159, 163, 165, 168, 170, 171, 174, 177, 284, 286, 290, 401, 484, 488, or 518.
[0166] In some embodiments, the FGF mutant is an FGF1 polypeptide consists of a polypeptide of SEQ ID NOs: 130, 131, 132, 133, 134, 135, 139, 141, 142, 143, 144, 146, 147, 148, 149, 150, 151, 152, 153, 156, 157, 158, 159, 163, 165, 168, 170, 171, 174, 177, 284, 286, 290, 401, 484, 488, or 518.
[0167] In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 80% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 85% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 90% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 91% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 92% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 93% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 94% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 95% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 96% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 97% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 98% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises about 99% or more sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant comprises 100% sequence identity to SEQ ID NO: 31. In some instances, the FGF mutant consists of SEQ ID NO: 31.
[0168] In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 80% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 85% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 90% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 91% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 92% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 93% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 94% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 95% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 96% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 97% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 98% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises about 99% or more sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant comprises 100% sequence identity to SEQ ID NO: 25785. In some instances, the FGF mutant consists of SEQ ID NO: 25785.
[0169] In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 80% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 85% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 90% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 91% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 92% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 93% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 94% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 95% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 96% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 97% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 98% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises about 99% or more sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant comprises 100% sequence identity to SEQ ID NO: 25851. In some instances, the FGF mutant consists of SEQ ID NO: 25851.
[0170] In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 80% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 85% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 90% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 91% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 92% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 93% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 94% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 95% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 96% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 97% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 98% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises about 99% or more sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant comprises 100% sequence identity to SEQ ID NO: 25880. In some instances, the FGF mutant consists of SEQ ID NO: 25880.
[0171] In some embodiments, the FGF mutant is an FGF1 polypeptide comprising about 80%, 85%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 80% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 85% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 90% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 91% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 92% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 93% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 94% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 95% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 96% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 97% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 98% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises about 99% or more sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant comprises 100% sequence identity to SEQ ID NO: 25923. In some instances, the FGF mutant consists of SEQ ID NO: 25923. [0172] In some embodiments, a FGF mutant described herein has a FGFR interaction activity that is equivalent to the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2 and a mitogenicity that is at least lower than the mitogenicity of the FGFl polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% to the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 70% to the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 75% to the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 80% to the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 85% to the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 90% to the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 95% to the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 96% to the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 97% to the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 98% to the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 99% to the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2. In some instances, the FGF mutant has a FGFR interaction activity that is about 100% to the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2.
[0173] In some instances, the FGF mutant has a FGFR interaction activity that is greater than the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 70%, 80%, 90%, or 100% greater than the FGFR interaction activity of FGFl polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 10% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 20% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 30% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 40% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 50% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 60% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 70% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 80% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a FGFR interaction activity that is about 90% greater than the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2.
[0174] In some instances, the FGF mutant has a mitogenicity that is at least 2-fold lower, 3-fold lower, 4-fold lower, 5-fold lower, 6-fold lower, 7-fold lower, 8-fold lower, 9-fold lower, or 10-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a mitogenicity that is at least 2-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a mitogenicity that is at least 3-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a mitogenicity that is at least 4-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a mitogenicity that is at least 5 -fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a mitogenicity that is at least 6-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a mitogenicity that is at least 7-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a mitogenicity that is at least 8-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a mitogenicity that is at least 9-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2. In some cases, the FGF mutant has a mitogenicity that is at least 10-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2.
[0175] In some embodiments, a FGF mutant described herein has a FGFR interaction activity that is equivalent to the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2 and a mitogenicity that is at least 1-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2.
[0176] In some embodiments, a FGF mutant described herein exhibits reduced or impaired mitogenic activity relative to a wild-type FGF polypeptide. In some instances, the FGF mutant is an FGF1 mutant. In some cases, an FGF1 mutant described herein exhibits reduced or impaired mitogenic activity relative to wild-type FGF1 polypeptide.
[0177] In some embodiments, a FGF mutant described herein exhibits enhanced stability relative to a wild-type FGF polypeptide. In some instances, the FGF mutant is an FGF1 mutant. In some cases, an FGF1 mutant described herein exhibits enhanced stability relative to wild-type FGF1 polypeptide.
[0178] In some embodiments, stability further comprises thermostability. In some embodiments, a FGF mutant described herein exhibits enhanced thermostability relative to a wild-type FGF polypeptide. In some instances, the FGF mutant is an FGFl mutant. In some cases, an FGFl mutant described herein exhibits enhanced thermostability relative to wild-type FGFl polypeptide.
[0179] In some embodiments, stability further comprises pH stability. In some embodiments, a FGF mutant described herein exhibits enhanced pH stability relative to a wild-type FGF polypeptide. In some instances, the FGF mutant is an FGF 1 mutant. In some cases, an FGF 1 mutant described herein exhibits enhanced pH stability relative to wild-type FGFl polypeptide.
[0180] In some embodiments, a FGF mutant described herein exhibits enhances FGF receptor interaction relative to a wild-type FGF polypeptide. In some instances, the FGF mutant is an FGFl mutant. In some cases, an FGF 1 mutant described herein exhibits enhanced FGF receptor interaction relative to wild-type FGFl polypeptide.
[0181] In some embodiments, a FGF mutant described herein promotes pre-adipocyte proliferation and differentiation. In some instances, the FGF mutant is an FGFl mutant. In some cases, an FGFl mutant described herein promotes pre-adipocyte proliferation and differentiation.
[0182] In some embodiments, a FGF mutant described herein promotes WAT remodeling and stimulates vascularization. In some instances, the FGF mutant is an FGFl mutant. In some cases, an FGFl mutant described herein promotes WAT remodeling and stimulates vascularization.
[0183] In some embodiments, a FGF mutant described herein has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof. In some instances, the FGF mutant is an FGFl mutant. In some cases, an FGFl mutant described herein has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof.
[0184] In some embodiments, a FGF mutant described herein has increased resistance to proteolysis relative to a wild-type FGF polypeptide. In some instances, the FGF mutant is an FGFl mutant. In some cases, an FGF 1 mutant described herein has increased resistance to proteolysis relative to a wild-type FGF polypeptide.
FGF Polypeptide Production
[0185] In some embodiments, FGF polypeptides described herein are generated recombinantly or are synthesized chemically. In some instances, FGF polypeptides described herein are generated recombinantly, for example, either by a host cell system, or in a cell-free system.
[0186] In some instances, FGF polypeptides described herein are generated recombinantly through a host cell system. In some cases, the host cell is a eukaryotic cell (e.g., mammalian cell, insect cells, yeast cells or plant cell) or a prokaryotic cell (e.g., gram -positive bacterium or a gram-negative bacterium).
[0187] In some embodiments, a eukaryotic host cell is a mammalian host cell. In some cases, a mammalian host cell is a stable cell line, or a cell line that has incorporated a genetic material of interest into its own genome and has the capability to express the product of the genetic material after many generations of cell division. In other cases, a mammalian host cell is a transient cell line, or a cell line that has not incorporated a genetic material of interest into its own genome and does not have the capability to express the product of the genetic material after many generations of cell division.
[0188] Exemplary mammalian host cells include 293T cell line, 293A cell line, 293FT cell line, 293F cells , 293 H cells, A549 cells, MDCK cells, CHO DG44 cells, CHO-S cells, CHO-K1 cells, Expi293F™ cells, Flp-In™ T-REx™ 293 cell line, Flp-In™-293 cell line, Flp-In™-3T3 cell line, Flp- In™-BHK cell line, Flp-In™-CHO cell line, Flp-In™-CV-l cell line, Flp-In™-Jurkat cell line, FreeStyle™ 293-F cells, FreeStyle™ CHO-S cells, GripTite™ 293 MSR cell line, GS-CHO cell line, HepaRG™ cells, T-REx™ Jurkat cell line, Per.C6 cells, T-REx™-293 cell line, T-REx™-CHO cell line, and T-REx™-HeLa cell line.
[0189] In some embodiments, a eukaryotic host cell is an insect host cell. Exemplary insect host cell include Drosophila S2 cells, Sf9 cells, Sf21 cells, High Five™ cells, and expresSF+® cells.
[0190] In some embodiments, a eukaryotic host cell is a yeast host cell. Exemplary yeast host cells include Pichia pastoris yeast strains such as GSl 15, KM71H, SMDl 168, SMDl 168H, and X-33, and Saccharomyces cerevisiae yeast strain such as INVScl .
[0191] In some embodiments, a eukaryotic host cell is a plant host cell. In some instances, the plant cells comprise a cell from algae. Exemplary plant cell lines include strains from
Chlamydomonas reinhardtii 137c, or Synechococcus elongatus PPC 7942.
[0192] In some embodiments, a host cell is a prokaryotic host cell. Exemplary prokaryotic host cells include BL21, Machl™, DH10B™, TOP10, DH5a, DHlOBac™, OmniMax™, MegaX™, DH12S™, INV110, TOP10F', INVaF, TOP10/P3, ccdB Survival, PIR1, PIR2, Stbl2™, Stbl3™, or Stbl4™.
[0193] In some instances, suitable polynucleic acid molecules or vectors for the production of a FGF polypeptide described herein include any suitable vectors derived from either a eukaryotic or prokaryotic sources. Exemplary polynucleic acid molecules or vectors include vectors from bacteria (e.g., E. coli), insects, yeast (e.g., Pichia pastoris), algae, or mammalian source. Bacterial vectors include, for example, pACYC177, pASK75, pBAD vector series, pBADM vector series, pET vector series, pETM vector series, pGEX vector series, pHAT, pHAT2, pMal-c2, pMal-p2, pQE vector series, pRSET A, pRSET B, pRSET C, pTrcHis2 series, pZA31-Luc, pZE21-MCS-l, pFLAG ATS, pFLAG CTS, pFLAG MAC, pFLAG Shift- 12c, pTAC-MAT-1, pFLAG CTC, or pTAC-MAT-2.
[0194] Insect vectors include, for example, pFastBacl, pFastBac DUAL, pFastBac ET, pFastBac HTa, pFastBac HTb, pFastBac HTc, pFastBac M30a, pFastBact M30b, pFastBac, M30c, pVL1392, pVL1393, pVL1393 M10, pVL1393 Mi l, pVL1393 M12, FLAG vectors such as pPolh-FLAGl or pPolh-MAT 2, or MAT vectors such as pPolh-MATl, or pPolh-MAT2. [0195] Yeast vectors include, for example, Gateway® pDEST™ 14 vector, Gateway® pDEST™ 15 vector, Gateway® pDEST™ 17 vector, Gateway® pDEST™ 24 vector, Gateway® pYES-DEST52 vector, pBAD-DEST49 Gateway® destination vector, pA0815 Pichia vector, pFLDl Pichi pastoris vector, pGAPZA, B, & C Pichia pastoris vector, p¥lC3.5K Pichia vector, pPIC6 A, B, & C Pichia vector, pPIC9K Pichia vector, pTEFl/Zeo, pYES2 yeast vector, pYES2/CT yeast vector, pYES2/NT A, B, & C yeast vector, or pYES3/CT yeast vector.
[0196] Algae vectors include, for example, pChlamy-4 vector or MCS vector.
[0197] Mammalian vectors include, for example, transient expression vectors or stable expression vectors. Exemplary mammalian transient expression vectors include p3xFLAG-CMV 8, pFLAG- Myc-CMV 19, pFLAG-Myc-CMV 23, pFLAG-CMV 2, pFLAG-CMV 6a,b,c, pFLAG-CMV 5.1, pFLAG-CMV 5a,b,c, p3xFLAG-CMV 7.1, pFLAG-CMV 20, p3xFLAG-Myc-CMV 24, pCMV- FLAG-MAT1, pCMV-FLAG-MAT2, pBICEP-CMV 3, or pBICEP-CMV 4. Exemplary mammalian stable expression vectors include pFLAG-CMV 3, p3xFLAG-CMV 9, p3xFLAG-CMV 13, pFLAG- Myc-CMV 21, p3xFLAG-Myc-CMV 25, pFLAG-CMV 4, p3xFLAG-CMV 10, p3xFLAG-CMV 14, pFLAG-Myc-CMV 22, p3xFLAG-Myc-CMV 26, pBICEP-CMV 1, or pBICEP-CMV 2.
[0198] In some instances, a cell -free system is used for the production of a FGF polypeptide described herein. In some cases, a cell-free system comprises a mixture of cytoplasmic and/or nuclear components from a cell and is suitable for in vitro nucleic acid synthesis. In some instances, a cell- free system utilizes prokaryotic cell components. In other instances, a cell -free system utilizes eukaryotic cell components. Nucleic acid synthesis is obtained in a cell-free system based on, for example, Drosophila cell, Xenopus egg, or HeLa cells. Exemplary cell-free systems include E. coli S30 Extract system, E. coli T7 S30 system, or PURExpress®.
Methods of Use
[0199] In some embodiments, a FGF mutant described herein is used for the treatment of a metabolic disease or condition. In some instances, the metabolic disease or condition is a FGF-related disease or condition. In some cases, the metabolic disease or condition is an FGFl -related disease or condition. In additional cases, the metabolic disease or condition is a FGF2 -related disease or condition. In some instances, the metabolic diseases or condition include, but are not limited to, diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), dyslipidemia, a cardiovascular disease, Hurler syndrome, Huntington's disease, Alzheimer's disease, Parkinson's disease, Crohn's disease, Fabry disease, or galactosemia. In additional instances, the metabolic disease or condition include impaired insulin sensitivity or glucose intolerance.
[0200] In some instances, a FGF mutant described herein is used for the treatment of a metabolic disease or condition, such as an FGFl -related disease or condition or a FGF2-related disease or condition. In some instances, a FGF mutant described herein is used for the treatment of an FGF1- related disease or condition.
[0201] In some instances, a FGF mutant described herein is used for the treatment of a metabolic disease or condition, such as diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), dyslipidemia, a cardiovascular disease, Hurler syndrome, Huntington's disease, Alzheimer's disease, Parkinson's disease, Crohn's disease, Fabry disease, or galactosemia.
[0202] In some instances, a FGF mutant described herein is used for the treatment of a metabolic disease or condition, such as impaired insulin sensitivity or glucose intolerance. In some cases, a FGF mutant described herein is used for the treatment of impaired insulin sensitivity. In some cases, a FGF mutant described herein is used for the treatment of glucose intolerance.
[0203] In some instances, administration of a FGF mutant described herein reduces blood glucose, improves insulin sensitivity, improves glucose tolerance, reduces systemic chronic inflammation, ameliorate hepatic steatosis, reduces food intake, or combinations thereof. In some cases, administration of a FGF mutant described herein reduces blood glucose. In some cases, administration of a FGF mutant described herein improves insulin sensitivity. In some cases, administration of a FGF mutant described herein improves glucose tolerance. In some cases, administration of a FGF mutant described herein reduces systemic chronic inflammation. In some cases, administration of a FGF mutant described herein ameliorate hepatic steatosis. In some cases, administration of a FGF mutant described herein reduces food intake.
Diabetes
[0204] In some embodiments, a metabolic disease or condition is diabetes (diabetes mellitus, DM). In some instances, diabetes is type 1 diabetes, type 2 diabetes, type 3 diabetes, type 4 diabetes, double diabetes, latent autoimmune diabetes (LAD), gestational diabetes, neonatal diabetes mellitus (NDM), maturity onset diabetes of the young (MODY), Wolfram syndrome, Alstrom syndrome, prediabetes, or diabetes insipidus. Type 2 diabetes, also called non-insulin dependent diabetes, is the most common type of diabetes accounting for 95% of all diabetes cases. In some instances, type 2 diabetes is caused by a combination of factors, including insulin resistance due to pancreatic beta cell dysfunction, which in turn leads to high blood glucose levels. In some cases, increased glucagon levels stimulate the liver to produce an abnormal amount of unneeded glucose, which contributes to high blood glucose levels.
[0205] Type 1 diabetes, also called insulin-dependent diabetes, comprises about 5% to 10% of all diabetes cases. Type 1 diabetes is an autoimmune disease where T cells attack and destroy insulin- producing beta cells in the pancreas. In some embodiments, Type 1 diabetes is caused by genetic and environmental factors. [0206] In some embodiments, the term double diabetes is used to describe patients diagnosed with both type 1 and 2 diabetes.
[0207] Type 4 diabetes is a recently discovered type of diabetes affecting about 20% of diabetic patients age 65 and over. In some embodiments, type 4 diabetes is characterized by age-associated insulin resistance.
[0208] In some embodiments, type 3 diabetes is used as a term for Alzheimer's disease resulting in insulin resistance in the brain.
[0209] LAD, also known as slow onset type 1 diabetes, is a slow developing form of type 1 diabetes where diagnosis frequently occurs after age 30. In some embodiments, LAD is further classified into latent autoimmune diabetes in adults (LAD A) or latent autoimmune diabetes in the young (LADY) or latent autoimmune diabetes in children (LADC).
[0210] Gestational diabetes is a type of diabetes that develops only during pregnancy and is characterized by high blood glucose levels due to insulin resistance. Gestational diabetes has a prevalence of about 9% and is frequently diagnosed during the third trimester of pregnancy. In some embodiments, gestational diabetes is further classified into type Al and A2. Gestational diabetes type Al is characterized by abnormal oral glucose tolerance test (OGTT) results but normal blood glucose levels during fasting and/or after meals; diet modification is sufficient to control blood glucose levels. Gestational diabetes type A2 is characterized by an abnormal OGTT results and abnormal blood glucose levels during fasting and/or after meals; insulin or other medications are required to control blood glucose levels.
[0211] Neonatal diabetes mellitus (NDM) is a rare type of diabetes affecting 1 in about 400,000 infants during the first 6 months of life. In some embodiments, NDM is further classified into permanent neonatal diabetes mellitus (PNDM) and transient neonatal diabetes mellitus (TNDM). PNDB is characterized by having insulin secretory failure during the late fetal or early post-natal period and does not go into remission. TNDM is characterized by having insulin secretory failure during the first few weeks of life but goes into remission in a few months. TNDM comprises about 50% to 60% of all NDM cases.
[0212] MODY, also known as monogenic diabetes, is a rare type of diabetes comprising about 1% to 2% of all diabetes cases. In some embodiments, MODY is caused by mutations in an autosomal dominant gene. In some instances, MODY if further classified into MODY1, MODY2, MODY3, MODY4, MODY5, MODY6, MODY7, MODY8, MODY9, MODY10, and MODY11.
[0213] Wolfram syndrome, also known as DIDMOAD (diabetes insipidus, diabetes mellitus, optic atrophy, and deafness) is a rare disorder affecting 1 in about 500,000 people. In some embodiments, Wolfram syndrome is further classified into Wolfram syndrome 1 (WFS 1) and Wolfram syndrome 2 (WFS2). In some instances, Wolfram syndrome is caused by mutations in the wolframin (WFS1) gene or CISD2 gene. [0214] Alstrom syndrome, also known as Alstrom-Halgren syndrome, is a rare disorder comprising about 500 to 900 reported cases worldwide. In some instances, Alstrom syndrome is caused by mutations in the ALMS1 gene. In some embodiments, Alstrom syndrome is characterized by type 2 diabetes occurring in early childhood, heart failure, vision problems, and obesity.
[0215] Prediabetes, also known as borderline diabetes, is a precursor stage to diabetes mellitus. In some cases, prediabetes is characterized by abnormal OGTT, fasting plasma glucose test, and hemoglobin AIC test results. In some embodiments, prediabetes is further classified into impaired fasting glycaemia or impaired fasting glucose (IFG) and impaired glucose tolerance (IGT). IFG is a condition in which blood glucose levels are higher than normal levels, but not elevated enough to be diagnosed as diabetes mellitus. IGT is a pre-diabetic state of abnormal blood glucose levels associated with insulin resistance and increased risk of cardiovascular pathology.
[0216] Diabetes insipidus (DI) is a rare disorder affecting 3 in about 100,000 people each year worldwide. In some embodiments, diabetes insipidus is further classified into neurogenic DI, nephrogenic DI, dipsogenic DI, and gestational DI. In some cases, neurogenic DI, also known as central DI, is characterized by damage to the pituitary gland, which results in disruption of antidiuretic hormone (ADH) storage and release. In some cases, nephrogenic DI is characterized by the inability of the kidneys to respond to ADH. In some cases, dipsogenic DI is characterized by a defect or damage in the thirst mechanism governed by the hypothalamus, which results in suppression of ADH and an increase in urine output. In some cases, gestational DI is characterized by degradation of vasopressin in the mother by placental vasopressinase.
[0217] In some embodiments, a FGF mutant described herein is used for the treatment of diabetes. In some instances, diabetes is type 1 diabetes, type 2 diabetes, type 3 diabetes, type 4 diabetes, double diabetes, latent autoimmune diabetes (LAD), gestational diabetes, neonatal diabetes mellitus (NDM), maturity onset diabetes of the young (MODY), Wolfram syndrome, Alstrom syndrome, prediabetes, or diabetes insipidus. In some cases, a FGF mutant described herein is used for the treatment of type 1 diabetes, type 2 diabetes, type 3 diabetes, type 4 diabetes, double diabetes, latent autoimmune diabetes (LAD), gestational diabetes, neonatal diabetes mellitus (NDM), maturity onset diabetes of the young (MODY), Wolfram syndrome, Alstrom syndrome, prediabetes, or diabetes insipidus. In some cases, a FGF mutant described herein is used for the treatment of type 1 diabetes. In some cases, a FGF mutant described herein is used for the treatment of type 2 diabetes. In some cases, a FGF mutant described herein is used for the treatment of type 3 diabetes. In some cases, a FGF mutant described herein is used for the treatment of type 4 diabetes. In some cases, a FGF mutant described herein is used for the treatment of double diabetes. In some cases, a FGF mutant described herein is used for the treatment of latent autoimmune diabetes (LAD) (e.g., LADA, LADY, or LADC). In some cases, a FGF mutant described herein is used for the treatment of gestational diabetes. In some cases, a FGF mutant described herein is used for the treatment of neonatal diabetes mellitus (NDM) (e.g., PNDM or TNDM). In some cases, a FGF mutant described herein is used for the treatment of maturity onset diabetes of the young (MODY) (e.g., MODY1, MODY2, MODY3, MODY4, MODY5, MODY6, MODY7, MODY8, MODY9, MODY10, or MODY11). In some cases, a FGF mutant described herein is used for the treatment of Wolfram syndrome (e.g., WFS 1 or WFS2). In some cases, a FGF mutant described herein is used for the treatment of Alstrom syndrome. In some cases, a FGF mutant described herein is used for the treatment of prediabetes. In some cases, a FGF mutant described herein is used for the treatment of diabetes insipidus (DI).
[0218] In some instances, the FGF mutant is an FGFl mutant. In some instances, an FGFl mutant described herein is used for the treatment of diabetes. In some instances, diabetes is type 1 diabetes, type 2 diabetes, type 3 diabetes, type 4 diabetes, double diabetes, latent autoimmune diabetes (LAD), gestational diabetes, neonatal diabetes mellitus (NDM), maturity onset diabetes of the young
(MODY), Wolfram syndrome, Alstrom syndrome, prediabetes, or diabetes insipidus. In some cases, an FGFl mutant described herein is used for the treatment of type 1 diabetes, type 2 diabetes, type 3 diabetes, type 4 diabetes, double diabetes, latent autoimmune diabetes (LAD), gestational diabetes, neonatal diabetes mellitus (NDM), maturity onset diabetes of the young (MODY), Wolfram syndrome, Alstrom syndrome, prediabetes, or diabetes insipidus. In some cases, an FGFl mutant described herein is used for the treatment of type 1 diabetes. In some cases, an FGFl mutant described herein is used for the treatment of type 2 diabetes. In some cases, an FGFl mutant described herein is used for the treatment of type 3 diabetes. In some cases, an FGFl mutant described herein is used for the treatment of type 4 diabetes. In some cases, an FGFl mutant described herein is used for the treatment of double diabetes. In some cases, an FGFl mutant described herein is used for the treatment of latent autoimmune diabetes (LAD) (e.g., LADA, LADY, or LADC). In some cases, an FGFl mutant described herein is used for the treatment of gestational diabetes. In some cases, an FGFl mutant described herein is used for the treatment of neonatal diabetes mellitus (NDM) (e.g., PNDM or TNDM). In some cases, an FGFl mutant described herein is used for the treatment of maturity onset diabetes of the young (MODY) (e.g., MODY1, MODY2, MODY3, MODY4, MODY5, MODY6, MODY7, MODY8, MODY9, MODY10, or MODY11). In some cases, an FGFl mutant described herein is used for the treatment of Wolfram syndrome (e.g., WFS 1 or WFS2). In some cases, an FGFl mutant described herein is used for the treatment of Alstrom syndrome. In some cases, an FGFl mutant described herein is used for the treatment of prediabetes. In some cases, an FGFl mutant described herein is used for the treatment of diabetes insipidus (DI).
Polycystic Ovary Syndrome
[0219] In some embodiments, a metabolic disease or condition is PCOS. PCOS is an endocrine disorder affecting about 5% to 10% of women in the United States. In some instances, PCOS is also known as hyperandrogenic anovulation (HA), Stein-Leventhal syndrome, or metabolic reproductive syndrome (MRS). In some cases, PCOS is characterized by enlarged ovaries that contain small collections of follicles, infrequent or prolonged menstrual periods, excess hair growth, acne, or obesity. In additional cases, PCOS is characterized by elevated levels of androgens such as testosterone. In some embodiments, PCOS is caused by a combination of genetic and environmental factors.
[0220] In some embodiments, a FGF mutant described herein is used for the treatment of PCOS. In some instances, the FGF mutant is an FGF 1 mutant. In some cases, an FGF 1 mutant described herein is used for the treatment of PCOS.
Metabolic Syndrome
[0221] In some embodiments, a metabolic disease or condition is metabolic syndrome (MetS). MetS is a group of conditions or risk factors comprising of abdominal obesity, high serum triglyceride levels, low high-density lipoprotein (HDL) levels, high fasting blood glucose levels, and high blood pressure. MetS affects about 34% of the population in the United States. In some embodiments, the risk associated with cardiovascular disease, diabetes, and stroke increases with MetS. In some cases, MetS is caused by obesity, insulin resistance, unhealthy lifestyle, or hormonal imbalance.
[0222] In some embodiments, a FGF mutant described herein is used for the treatment of metabolic syndrome. In some instances, the FGF mutant is an FGF1 mutant. In some cases, an FGF1 mutant described herein is used for the treatment of metabolic syndrome.
Obesity
[0223] In some embodiments, a metabolic disease or condition is obesity. Obesity is a medical condition involving excess body fat. Obesity affects about 34.9% of the adult population in the United States and about 17% of the child population in the United States. In some instances, obesity is further classified as class I obesity, class II obesity, and class III obesity. In some embodiments, a body mass index (BMI) or Quetelet index value is utilized to diagnose obesity. In some
embodiments, class I obesity is characterized by BMI values ranging between 30 kg/m2 and 35 kg/m2. In some embodiments, class II obesity is characterized by BMI values ranging between 35 kg/m2 and 40 kg/m2. In some embodiments, class I obesity is characterized by BMI values above 40 kg/m2.
[0224] In some embodiments, a FGF mutant described herein is used for the treatment of obesity. In some instances, the FGF mutant is an FGF 1 mutant. In some cases, an FGF 1 mutant described herein is used for the treatment of obesity.
Non-Alcoholic Fatty Liver Disease
[0225] In some embodiments, a metabolic disease or condition is non-alcoholic fatty liver disease (NAFLD). NAFLD is one of the most common causes of chronic liver disease affecting about 3% of the population in the United States. In some embodiments, NAFLD is further classified into hepatic steatosis and non-alcoholic steatohepatitis (NASH). Hepatic steatosis, also known as simple fatty liver, is characterized by a net retention of fat within hepatocytes resulting from an imbalance of uptake of fat and its oxidation and export. NASH is also characterized by a high fat retention in hepatocytes, but is also characterized by inflammation and hepatocyte damage. In some
embodiments, NASH is further classified into three stages: fatty liver, liver fibrosis, and cirrhosis. The fatty liver stage comprises a large accumulation of fat within hepatocytes, which leads to liver enlargement. Liver fibrosis is characterized by formation of scar tissue and continued hepatocyte injury. The final stage, cirrhosis, is characterized by late stage fibrosis that blocks the portal blood flow and thus, impairs normal function. In some embodiments, the risk associated with NASH increases with insulin resistance, release of cytokines by adipocytes, or oxidative stress in
hepatocytes.
[0226] In some embodiments, a FGF mutant described herein is used for the treatment of NAFLD. In some instances, the FGF mutant is an FGF 1 mutant. In some cases, an FGF 1 mutant described herein is used for the treatment of NAFLD.
Dyslipidemia
[0227] In some embodiments, a metabolic disease or condition is dyslipidemia. Dyslipidemia is characterized by abnormal levels of lipids, such as triglycerides and cholesterol, and lipoproteins or chylomicrons, in the blood. In some embodiments, a common type of dyslipidemia is hyperlipidemia, which is characterized by elevated levels of any or all lipids and/or lipoproteins in the blood. In some embodiments, hyperlipidemia is further classified into familial or primary hyperlipidemia and acquired or secondary hyperlipidemia. Familial hyperlipoproteinemia is caused by specific genetic abnormalities. In some instances, familial hyperlipidemia is further classified into
hyperlipoproteinemia type I, II, III, IV, and V. In some cases, hyperlipoproteinemia type II is further classified into type Ila and lib. Secondary hyperlipoproteinemia is caused by another underlying disorder that affects lipoprotein metabolism and plasma lipid levels. In some embodiments, secondary hyperlipoproteinemia is caused by diabetes mellitus, hypothyroidism, renal failure, nephrotic syndrome, alcohol usage, or endocrine or metabolic syndromes.
[0228] In some embodiments, a FGF mutant described herein is used for the treatment of dyslipidemia. In some instances, dyslipidemia is hyperlipidemia. In some instances, the FGF mutant is an FGF 1 mutant. In some cases, an FGF 1 mutant described herein is used for the treatment of dyslipidemia. In some cases, an FGF1 mutant described herein is used for the treatment of hyperlipidemia.
Insulin Resistance [0229] In some embodiments, a metabolic disease or condition is insulin resistance. Insulin resistance, also known as insulin insensitivity, is a medical condition in which cells fail to respond to the hormone insulin and are unable to properly absorb glucose from the bloodstream, elevating blood glucose levels, which in turn leads to an increased production of insulin by pancreatic beta cells. In some instances, the risk associated with insulin resistance increases with genetic factors, such as insulin receptor mutations or LMNA mutations, or physiological factors, such as obesity, hypertension, or hypertriglyceridemia. In some instances, insulin resistance increases the risk for developing type 2 diabetes and prediabetes.
[0230] In some embodiments, a FGF mutant described herein is used for the treatment of insulin resistance. In some instances, the FGF mutant is an FGF1 mutant. In some cases, an FGF1 mutant described herein is used for the treatment of insulin resistance.
Lipodystrophy
[0231] In some embodiments, a metabolic disease or condition is lipodystrophy. Lipodystrophy is a disorder of adipose tissue characterized by abnormal and selective loss of adipose tissue from different parts of the body. In some instances, lipodystrophy is further classified into congenital lipodystrophy and acquired lipodystrophy. In some cases, congenital lipodystrophy is caused by a genetic mutation and some types include congenital generalized lipodystrophy, also known as Beradinelli-Seip syndrome, and familial partial lipodystrophy. In some embodiments, acquired lipodystrophy includes acquired partial lipodystrophy, acquired generalized lipodystrophy, centrifugal abdominal lipodystrophy, also known as lipodystrophia centrifugalis abdominalis infantilis, lipoatrophia annularis, also known as Ferreira-Marques lipoatrophia, localized lipodystrophy, or HIV- associated lipodystrophy.
[0232] In some embodiments, a FGF mutant described herein is used for the treatment of lipodystrophy. In some instances, the FGF mutant is an FGF1 mutant. In some cases, an FGF1 mutant described herein is used for the treatment of lipodystrophy.
Cardiovascular Disease
[0233] In some embodiments, a metabolic disease or condition is a cardiovascular disease.
Cardiovascular disease (CVD) encompasses a range of conditions affecting the heart; about 31% of global deaths are caused by cardiovascular disease. In some cases, cardiovascular disease comprises hypertension, coronary artery diseases (CAD), stroke such as hemorrhagic stroke or ischemic stroke, rheumatic heart disease, hypertensive heart disease, cardiomyopathy, heart arrhythmia, congenital heart disease, carditis, aortic aneurysms, renal artery stenosis, peripheral artery disease, venous thrombosis, heart valve disease, deep vein thrombosis, cerebrovascular disease, cardiac dysrhythmia, chronic obstructive pulmonary disease, or inflammatory heart disease such as myocarditis. In some instances, the risk associated with CVD increases with behavioral factors such as unhealthy diet, physical inactivity, tobacco use, and harmful use of alcohol. In some cases, a major precursor of CVD is atherosclerosis.
[0234] In some embodiments, a FGF mutant described herein is used for the treatment of a cardiovascular disease. In some cases, cardiovascular disease comprises hypertension, coronary artery diseases (CAD), stroke such as hemorrhagic stroke or ischemic stroke, rheumatic heart disease, hypertensive heart disease, cardiomyopathy, heart arrhythmia, congenital heart disease, carditis, aortic aneurysms, renal artery stenosis, peripheral artery disease, venous thrombosis, heart valve disease, deep vein thrombosis, cerebrovascular disease, cardiac dysrhythmia, chronic obstructive pulmonary disease, or inflammatory heart disease such as myocarditis. In some cases, a FGF mutant described herein is used for the treatment of hypertension, coronary artery diseases (CAD), stroke such as hemorrhagic stroke or ischemic stroke, rheumatic heart disease, hypertensive heart disease, cardiomyopathy, heart arrhythmia, congenital heart disease, carditis, aortic aneurysms, renal artery stenosis, peripheral artery disease, venous thrombosis, heart valve disease, deep vein thrombosis, cerebrovascular disease, cardiac dysrhythmia, chronic obstructive pulmonary disease, or
inflammatory heart disease such as myocarditis. In some cases, a FGF mutant is used for the treatment of hypertension. In some cases, a FGF mutant is used for the treatment of coronary artery diseases (CAD).
[0235] In some instances, the FGF mutant is an FGF1 mutant. In some cases, an FGF1 mutant described herein is used for the treatment of a cardiovascular disease. In some cases, cardiovascular disease comprises hypertension, coronary artery diseases (CAD), stroke such as hemorrhagic stroke or ischemic stroke, rheumatic heart disease, hypertensive heart disease, cardiomyopathy, heart arrhythmia, congenital heart disease, carditis, aortic aneurysms, renal artery stenosis, peripheral artery disease, venous thrombosis, heart valve disease, deep vein thrombosis, cerebrovascular disease, cardiac dysrhythmia, chronic obstructive pulmonary disease, or inflammatory heart disease such as myocarditis. In some cases, an FGF1 mutant described herein is used for the treatment of hypertension, coronary artery diseases (CAD), stroke such as hemorrhagic stroke or ischemic stroke, rheumatic heart disease, hypertensive heart disease, cardiomyopathy, heart arrhythmia, congenital heart disease, carditis, aortic aneurysms, renal artery stenosis, peripheral artery disease, venous thrombosis, heart valve disease, deep vein thrombosis, cerebrovascular disease, cardiac dysrhythmia, chronic obstructive pulmonary disease, or inflammatory heart disease such as myocarditis. In some cases, an FGF1 mutant is used for the treatment of hypertension. In some cases, an FGF1 mutant is used for the treatment of coronary artery diseases (CAD).
Additional Therapeutic Agents [0236] In some embodiments, disclosed herein include use of a FGF mutant (e.g., FGF1 mutant) in combination with an additional therapeutic agent. In some instances, the additional therapeutic agent is a therapeutic agent for the treatment of a metabolic disease selected from diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic steatohepatitis (NASH), nonalcoholic fatty liver disease (NAFLD), dyslipidemia, a cardiovascular disease, Hurler syndrome, Huntington's disease, Alzheimer's disease, Parkinson's disease, Crohn's disease, Fabry disease, and galactosemia. In some cases, a FGF mutant (e.g., FGF1 mutant) described herein is used in combination with a therapeutic agent for the treatment of a metabolic disease selected from diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic
steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), dyslipidemia, a cardiovascular disease, Hurler syndrome, Huntington's disease, Alzheimer's disease, Parkinson's disease, Crohn's disease, Fabry disease, and galactosemia.
[0237] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of diabetes. In some instances, the therapeutic agent comprises a therapeutic agent from thiazolidinedione (or glitazones) class of therapeutic agents. In some instances, thiazolidinedione comprises pioglitazone, rosiglitazone, lobeglitazone, and troglitazone. In some instances, additional therapeutic agents for the treatment of diabetes includes: acarbose, acetohexamide, albiglutide, alogliptin or alogliptin benzoate, the combination of alogliptin and metformin, the combination of alogliptin and pioglitazone, anagliptin, bromocriptine mesylate, canaglifozin, the combination of canagliflozin and metformin, carbutamide, chlorpropamide, ciglitazone, colesevelam, dapagliflozin or dapaglifozin propanediol, the combination of dapagliflozin and metformin, darglitazone, dulaglutide, dutogliptin, empagliflozin, the combination of empagliflozin and linagliptin, the combination of empagliflozin and metformin, englitazone, exenatide, gemigliptin, glibenclamide, glibornuride, gliclazide, glimepiride, glipizide, the combination of glipizide and metformin, glyburide, gliquidone, glisoxepide, glyburide micronized, the combination of glyburide and metformin, glyclopyramide, insulin, insulin aspart, insulin aspart protamine, the combination of insulin aspart and insulin aspart protamine, insulin degludec, insulin detemir, insulin glargine, insulin glulisine, insulin human isophane or insulin isophane, insulin lispro, insulin lispro protamine, the combination of insulin lispro and lispro protamine, insulin neutral protamine Hagedorn (NPH), the combination of insulin NPH and insulin isophane, the combination of insulin NPH and insulin, linagliptin, the combination of linagliptin and metformin, liraglutide, lixisenatide, metformin or metformin HC1, mifepristone, miglitol, mitiglinide, nateglinide, netoglitazone, omarigliptin, the combination of pioglitazone and glimepiride, the combination of pioglitazone and metformin, pramlintide or pramlintide acetate, repaglinide, the combination of repaglinide and metformin, rivoglitazone, rosiglitazone or rosiglitazone maleate, the combination of rosiglitazone and glimepiride, the combination of rosiglitazone and metformin, saxagliptin, the combination of saxagliptin and metformin, or sitagliptin or sitagliptin phosphate, the combination of sitagliptin phosphate and metformin, taspoglutide, teneligliptin, tolazamide, tolbutamide, or trelagliptin.
[0238] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of polycystic ovarian syndrome (PCOS) and includes: estrogen, progestin, a combination of estrogen and progestin, spironolactone, clomiphene, eflornithine, or gonadotropin hormones such as luteinizing hormone (LH) or follicle stimulating hormone (FSH). In some embodiments, a therapeutic agent is used to treat obesity and includes: orlistat, lorcaserin, topiramate, phentermine, the combination of phentermine and topiramate, diethylpropion, phendimetrazine, benzphetamine, sibutramine, rimonabant, phenylpropanolamine, or the combination of fenfluramine and phentermine.
[0239] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of non-alcoholic steatohepatitis (NASH) and includes: obeticholic acid, or elafibranor or GFT505 (Genfit S.A.). In some embodiments, a therapeutic agent is used to treat non-alcoholic fatty liver disease (NAFLD) and includes: pentoxifylline or oxpentifylline.
[0240] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of dyslipidemia and includes: bile acid sequestrants such as cholestyramine resin or colestipol; cholesterol absorption inhibitors such as exetimibe; fibrates such as fenofibrate or fenofibric acid; lipid-regulating agents such as gemfibrozil, icosapent ethyl, or omega-3-acid ethyl esters; microsomal tryglyceride transfer protein (MTP) inhibitors such as lomitapide mesylate;
nicotinic acid derivatives such as niacin; oligonucleotide inhibitors such as mipomersen; proprotein convertase subtilisin kexin type 9 (pcsk9) inhibitors such as alirocumab or evolocumab; statins (3- hydroxy-3 -methyl -glutaryl-CoA (HMG-COA) reductase inhibitors) such as atorvastatin, fluvastatin, lovastatin, pitavastatin, pravastatin, rosuvastatin, or simvastatin; statins in combination with calcium channel blockers such as atorvastatin and amlopidine; statins in combination with cholesterol absorption inhibitors such as simvastatin and ezetimibe; or statins in combination with nicotinic acid derivatives such as lovastatin and niacin or simvastatin and niacin.
[0241] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of cardiovascular disease and includes: aldosterone inhibitors such as eplerenone, spironolactone, the combination of eplerenone and potassium-sparing diuretics; angiotensin converting ezyme (ACE) inhibitors such as quinapril, perindopril, Ramipril, captopril, benazepril, trandolapril, fosinopril, lisinopril, moexipril, or enalapril; angiotensin II receptor blocker (ARBs) such as candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, the combination of irbesartan and hydrochlorothiazide, or the combination of losartan and hydrochlorothiazide; beta- blockers such as acebutolol, atenolol, betaxolol, bisoprolol, carteolol, carvedilol, esmolol, labetalol, metoprolol, nadolol, nebivolol, pentutolol, pindalol, propranolol, sotalol, or timolol; calcium channel blockers such as amlopidine, the combination of amlodipine and atorvastatin, valsartan, telmisartan, olmesartan, olmesartan and hydrochlorothiazide, aliskiren, aliskiren and hydrochlorothiazide, or perindopril, diltiazem, felodipine, isradipine, nicardipine, nifedipine, nimodipine, nisolipine, verapamil, vidipine, dihydropyridine, phenylalkylamine, benzodiazepine, mibefradil, bepridil, flunarizine, fluspirilene, fendiline, gabapentin, pregabalin, or ziconotide; cholesterol lowering drugs such as statins (see therapeutic agents for dyslipidemia, supra), or niacin; digoxin; diuretics such as chlorothiazide, hydrochlorothiazide, indapamide, metolazone, chlorthalidone, bumetanide, ethacrynic acid, furosemide, torsemide, amiloride, eplerenone, spironolactone, or triamterene; supplements such as potassium or magnesium; or vasodilators such as nitroglycerin, alprostadil, doxazosin, prazosin, terazosin, clonidine, hydralazine, or minoxidil.
[0242] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of Hurler's syndrome and includes: laronidase, or delivery of the gene iduronidase.
[0243] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of Huntington's disease and includes: tetrabenazine, haloperidol, chlorpromazine, risperidone, amantadine, levetiracetam, citalopram, fluoxetine, sertraline, quetiapine, olanzapine, valproate, carbamazepine, or lamotrigine.
[0244] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of Alzheimer's disease and includes: galantamine, rivastigmine, donepezil, memantine, or the combination of donepezil and memantine.
[0245] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of Parkinson's disease and includes: levodopa and carbidopa; dopamine agonists such as ropinirole HCl, pramipexodole dihydrochloride, rotigotine; amantadine; anticholinergics such as trihexyphenidyl or benztropine mesylate; selegiline HCl or rasagiline mesylate; or catechol-O-methyl transferase (COMT) inhibitors such as entacapone, tolcapone, or nitecapone.
[0246] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of Crohn's disease and includes: 5 -aminosalicylates such as sulfasalazine or mesalamine; corticosteroids such as prednisone, methylprednisolone, hydrocortisone, or budesonide; immune system suppressors such as azathioprine, mercaptopurine, infliximab, adalimumab, certolizumab pegol, methotrexate, cyclosporine, tracrolimus, natalizumab, vedolizumab, or ustekinumab;
antibiotics such as metronidazole or ciprofloxacin; other medications such as psyllium powder, methylcellulose, acetaminophen, ibuprofen, or naproxen sodium.
[0247] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of Fabry disease and includes: agalsidase beta or alpha-galactosidase.
[0248] In some embodiments, the additional therapeutic agent is a therapeutic agent for the treatment of Fabry disease and includes: agalsidase beta or alpha-galactosidase.
[0249] In some embodiments, the additional therapeutic agent is a glucagon-like peptide- 1 receptor
(GLP-1) agonist (or incretin mimetics). In some instances, exemplary GLP-1 agonists include, but are not limited to, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, semaglutide, or taspoglutide.
[0250] In some embodiments, the additional therapeutic agent is an inhibitor of dipeptidyl peptidase 4 (DPP-4 inhibitor or gliptin). In some instances, exemplary DPP-4 inhibitors include, but are not limited to, alogliptin, anagliptin, berberine, dutogliptin, gemigliptin, linagliptin, lupeol, omarigliptin, saxagliptin, sitagliptin, teneligliptin, trelagliptin, or vildagliptin.
[0251] In some embodiments, the additional therapeutic agent is a sodium-glucose co -transporter 2 (SGLT2) inhibitor. In some instances, exemplary SGLT2 inhibitors include, but are not limited to, canagliflozin, dapagliflozin, or empagliflozin.
[0252] In some embodiments, the additional therapeutic agent is metformin. In some
embodiments, the additional therapeutic agent is metformin HC1. In some instances, a FGF mutant described herein is used in combination with metformin. In some instances, a FGF mutant described herein is used in combination with metformin HC1.
FGF Polypeptide Conjugation
[0253] In some embodiments, a FGF mutant described herein is further attached to a conjugating moiety. In some instances, the attachment is a covalent attachment. In other instances, the attachment is a non-covalent attachment. In additional instances, the attachment is an irreversible conjugation. In some cases, a conjugating moiety is attached to the N-terminus, the C-terminus, or at an internal site on FGF.
[0254] In some cases, a FGF mutant described herein is further modified (e.g., by a post- translational modification) and a conjugating moiety is attached to a modified site (e.g., a post- translationally modified site). In some instances, examples of post-translational modification include myristoylation, palmitoylation, isoprenylation (or prenylation) (e.g., farnesylation or
geranylgeranylation), glypiation, acylation (e.g., O-acylation, N-acylation, S-acylation), alkylation (e.g., additional of alkyl groups such as methyl or ethyl groups), amidation, glycosylation, hydroxylation, iodination, nucleotide addition, oxidation, phosphorylation, succinylation, sulfation, glycation, carbamylation, glutamylation, or deamidation. In some cases, a FGF mutant described herein is modified by a post-translational modification. In some cases, a FGF mutant described herein is modified by myristoylation, palmitoylation, isoprenylation (or prenylation) (e.g., farnesylation or geranylgeranylation), glypiation, acylation (e.g., O-acylation, N-acylation, S-acylation), alkylation (e.g., additional of alkyl groups such as methyl or ethyl groups), amidation, glycosylation, hydroxylation, iodination, nucleotide addition, oxidation, phosphorylation, succinylation, sulfation, glycation, carbamylation, glutamylation, or deamidation.
[0255] In some instances, a conjugating moiety comprises a polymer, a protein or its functional fragments thereof, a conjugating peptide, an antibody or its binding fragments thereof, or an antibody mimetic. In some instances, a FGF conjugate comprises a FGF-polymer conjugate. In some instances, a FGF conjugate comprises a FGF-protein conjugate. In some instances, a FGF conjugate comprises a FGF-conjugating peptide conjugate. In some instances, a FGF conjugate comprises a FGF-antibody conjugate. In some instances, a FGF conjugate comprises a FGF-antibody mimetic conjugate.
[0256] In some cases, a conjugating moiety (or conjugate) described below alters one or more properties of a FGF polypeptide. In some instances, the one or more properties include biological properties and/or physicochemical properties. Exemplary biological properties and/or
physicochemical properties include, but are not limited to, pharmacokinetics, pharmacodynamics, metabolic stability, biodistribution, water solubility, lipophilicity, or tissue targeting capabilities. In some instances, a conjugating moiety (or conjugate) alters pharmacokinetics, pharmacodynamics, metabolic stability, biodistribution, water solubility, lipophilicity, tissue targeting capabilities, or a combination thereof, of a FGF polypeptide. In some instances, a conjugating moiety (or conjugate) enhances pharmacokinetics, pharmacodynamics, metabolic stability, biodistribution, water solubility, lipophilicity, tissue targeting capabilities, or a combination thereof, of a FGF conjugate relative to an unconjugated FGF polypeptide. In some cases, a conjugating moiety (or conjugate) enhances the in vivo half-life, in vivo potency, activity, or a combination thereof, of a FGF conjugate relative to an unconjugated FGF polypeptide. In some cases, a conjugating moiety (or conjugate) facilitates and/or enables transport of a FGF polypeptide described herein across the blood-brain barrier (BBB).
Polymers
[0257] In some embodiments, a conjugating moiety comprises a polymer. In some instances, a polymer is a natural or synthetic polymer, consisting of, e.g., long chains of branched or unbranched monomers, and/or cross-linked network of monomers in two or three dimensions. In some cases, a polymer comprises a water soluble polymer or a water insoluble polymer. In some instances, a FGF polypeptide conjugate comprises a FGF-polymer conjugate. In some cases, a polymer conjugate alters one or more properties (e.g., biological properties and/or physicochemical properties) of a FGF polypeptide.
Water Soluble Polymers
[0258] In some embodiments, a polymer is a water soluble polymer. Exemplary water soluble polymers include polyalkylen oxide (e.g., polyethylene glycol), polysaccharide, polyoxazoline polymer, polyacrylic acid or polyamine. In some instances, a water soluble polymer comprises polyalkylen oxide (e.g., polyethylene glycol). In some embodiments, a polyalkylene oxide (e.g., PEG) is a polydisperse or monodisperse compound. In some instances, a polydispersed PEG comprises disperse distribution of different molecular weight of the material, characterized by mean weight (weight average) size and dispersity. In some instances, a monodisperse PEG comprises one size of PEG molecules.
[0259] In some embodiments, the molecular weight of the polyalkylene oxide (e.g., PEG) is about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1450, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3250, 3350, 3500, 3750, 4000, 4250, 4500, 4600, 4750, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 10,000, 12,000, 20,000, 35,000, 40,000, 50,000, 60,000, or 100,000 Da.
[0260] In some instances, a FGF polypeptide described herein is conjugated or bond to a PEG molecule. In some instances, a FGF polypeptide described herein is conjugated or bond to a PEG molecule of molecule weight of about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1450, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3250, 3350, 3500, 3750, 4000, 4250, 4500, 4600, 4750, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 10,000, 12,000, 20,000, 35,000, 40,000, 50,000, 60,000, or 100,000 Da. In some cases, a FGF polypeptide selected from SEQ ID NOs: 3-25917 and 25922-25974 is conjugated or bond to a PEG molecule, e.g., with a PEG molecule weight of about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1450, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3250, 3350, 3500, 3750, 4000, 4250, 4500, 4600, 4750, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 10,000, 12,000, 20,000, 35,000, 40,000, 50,000, 60,000, or 100,000 Da.
[0261] In some instances, modification of a target molecule of interest (e.g., a FGF polypeptide) by PEG is referred to as PEGylation. In reference to a FGF polypeptide described herein, PEG is sometimes covalently attached to a FGF polypeptide. In other cases, PEG is non-covalently attached a FGF polypeptide. In some instances, suitable functional groups involved in conjugation chemistry include reactive side chains of lysine, cysteine, histidine, arginine, aspartic acid, glutamic acid, serine, threonine, or tyrosine; N-terminal amino group; or the C-terminal carboxylic acid. In some instances, a PEG molecule is further modified with a bifunctional linker (e.g., homobifunctional or
heterobifunctional), which reacts with one or more of a reactive group of a FGF polypeptide.
[0262] In some instances, PEGylation is an enzyme-mediated PEGylation. In such cases, an enzyme-mediated PEGylation involves, for example, transglutaminase which catalyzes the transfer of an amino-PEG to the acyl group in the side chain of a glutamine; GlycoPEGylation, which utilizes O- GalNAc-transferase and a sialytransferase to couple PEG in a site-specific manner to specific serine or threonine residues, thereby mimicking O-glycosylation; or sortase A mediated PEGylation, which attaches a polymer to the C-terminus of a polypeptide through a LPXTG motif (SEQ ID NO: 25918). In some instances, an enzyme-mediated PEGylation is GlycoPEGylation. In some instances, an enzyme-mediated PEGylation is a GlycoPEGylation as described in U.S. Pub. Nos. 2008/0146782, and 2010/0286067; and U.S. Patent Nos. 8,063,015 and 8,911,967. In some instances, an enzyme- mediated PEGylation is a GlycoPEGylation as described in DeFrees et al, "GlycoPEGylation of recombinant therapeutic proteins produced in Escherichia coli," Glycobiology 16(9): 833-843, 2006.
[0263] In some embodiments, a FGF polypeptide described herein is modified by PEGylation. In some instances, a FGF polypeptide described herein is modified by an enzyme -mediated PEGylation. In some cases, a FGF polypeptide described herein is modified by GlycoPEGylation. In some instances, a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974 is modified by PEGylation. In some instances, a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974 is modified by GlycoPEGylation.
[0264] In some instances, a water soluble polymer comprises a polysaccharide. Exemplary polysaccharides include, but are not limited to, dextran, polysialic acid (PSA), hyaluronic acid (HA), amylose, heparin, heparan sulfate (HS), dextrin, or hydroxyethyl-starch (HES). In some instances, a FGF-polymer conjugate comprises a FGF-polysaccharide conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-dextran conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-PSA conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-HA conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-amylose conjugate. In some cases, a FGF- polymer conjugate comprises a FGF-heparin conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-heparan sulfate conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-dextrin conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-HES conjugate. In some cases, a FGF-polysaccharide conjugate comprises a FGF polypeptide of SEQ ID NOs: 3- 25917 and 25922-25974.
[0265] In some embodiments, modification of a FGF polypeptide described herein by
hydroxyethyl-starch (HES) is referred to as HESylation. In some instances, HESylation is an enzyme-mediated conjugation, in which transglutaminase (Tgase) catalyzes the transfer of a primary amino group to the acyl group of glutamine. In some instances, the conjugation occurs between the primary amino group (e.g., the primary group of a lysine residue) of the polypeptide of interest to the acyl group of glutamine in HES. In other instances, the HES group is modified to contain an amino- HES derivative and the conjugation occurs between the primary amino group of the amino-HES derivative and an acyl group of the polypeptide of interest. In some instances, a FGF polypeptide described herein is modified by HESylation. In some instances, a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974 is modified by HESylation.
[0266] In some embodiments, a water soluble polymer comprises a polyoxazoline polymer. A polyoxazoline polymer is a linear synthetic polymer, and similar to PEG, comprises a low
polydispersity. In some instances, a polyoxazoline polymer is a polydispersed polyoxazoline polymer, characterized with an average molecule weight. In some cases, the average molecule weight of a polyoxazoline polymer includes, for example, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 10,000, 12,000, 20,000, 35,000, 40,000, 50,000, 60,000, 100,000, 200,000, 300,000, 400,000, or 500,000 Da.
[0267] In some instances, a FGF-polymer conjugate comprises a FGF-polyoxazoline polymer conjugate. In some instances, a polyoxazoline polymer comprises poly(2-methyl 2-oxazoline) (PMOZ), poly(2-ethyl 2-oxazoline) (PEOZ), or poly(2 -propyl 2-oxazoline) (PPOZ). In some cases, a FGF-polymer conjugate comprises a FGF-PMOZ conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-PEOZ conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-PPOZ conjugate. In some cases, a FGF-polyoxazoline polymer conjugate comprises a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
[0268] In some instances, a water soluble polymer comprises a polyacrylic acid polymer. In some instances, a FGF-polymer conjugate comprises a FGF-polyacrylic acid polymer. In some cases, a FGF-polyacrylic acid polymer conjugate comprises a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
[0001] In some instances, a water soluble polymer comprises polyamine. Polyamine is an organic polymer comprising two or more primary amino groups. In some embodiments, a polyamine includes a branched polyamine, a linear polyamine, or cyclic polyamine. In some cases, a polyamine is a low- molecular-weight linear polyamine. Exemplary polyamines include putrescine, cadaverine, spermidine, spermine, ethylene diamine, 1,3-diaminopropane, hexamethylenediamine,
tetraethylmethylenediamine, and piperazine. In some instances, a FGF-polymer conjugate comprises a FGF-polyamine conjugate. In some instances, a FGF-polymer conjugate comprises a FGF- polyamine conjugate in which the polyamine comprises putrescine, cadaverine, spermidine, spermine, ethylene diamine, 1,3-diaminopropane, hexamethylenediamine, tetraethylmethylenediamine, or piperazine. In some instances, a FGF-polymer conjugate comprises a FGF-putre seine conjugate. In some instances, a FGF-polymer conjugate comprises a FGF-cadaverine conjugate. In some instances, a FGF-polymer conjugate comprises a FGF-spermidine conjugate. In some instances, a FGF-polymer conjugate comprises a FGF-spermine conjugate. In some instances, a FGF-polymer conjugate comprises a FGF-ethylene diamine conjugate. In some instances, a FGF-polymer conjugate comprises a FGF- 1,3-diaminopropane conjugate. In some instances, a FGF-polymer conjugate comprises a FGF- hexamethylenediamine conjugate. In some instances, a FGF-polymer conjugate comprises a FGF- tetraethylmethylenediamine conjugate. In some instances, a FGF-polymer conjugate comprises a FGF- piperazine conjugate. In some cases, a FGF-polysaccharide conjugate comprises a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
[0269] In some embodiments, modification of a FGF polypeptide described herein by a polyamine is referred to as polyamination. In some instances, polyamination is an enzyme -mediated conjugation, in which transglutaminase (Tgase) catalyzes the transfer of a primary amino group to an acyl group. In some instances, polyamination is as described in Zhang, et al, "Synthesis and applications of polyamine amino acid residues: improving the bioactivity of an analgesic neuropeptide, Neurotensin," J Med Chem 52(6): 1514-1517 (2009). In some instance, a FGF polypeptide described herein is modified by polyamination. In some instances, a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974 is modified by polyamination.
Water Insoluble Polymers
[0270] In some embodiments, a polymer is a water insoluble polymer. Exemplary water insoluble polymers include, but are not limited to, polyphosphazines, poly(vinyl alcohols), polyamides, polycarbonates, polyalkylenes, polyacrylamides, polyalkylene glycols, polyalkylene terephthalates, polyvinyl ethers, polyvinyl esters, polyvinyl halides, polyvinylpyrrolidone, polyglycolides, polysiloxanes, polyurethanes, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate) polyethylene, polypropylene, poly (ethylene terephthalate), poly(vinyl acetate), polyvinyl chloride, polystyrene, polyvinyl pyrrolidone, pluronics and polyvinylphenol and copolymers thereof.
[0271] In some instances, a water insoluble polymer further comprises a synthetically modified natural polymer, which includes, for example, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxybutyl methyl cellulose, cellulose acetate, cellulose propionate, cellulose acetate butyrate, cellulose acetate phthalate, carboxymethyl cellulose, cellulose triacetate, cellulose sulfate sodium salt, or polymers of acrylic and methacrylic esters and alginic acid.
[0272] In some instances, a water insoluble polymer also comprises a biodegradable polymer. In some instances, biodegradable polymers comprise polylactides, polyglycolides and copolymers thereof, poly(ethylene terephthalate), poly(butyric acid), poly(valeric acid), poly(lactide-co- caprolactone), poly(lactide-co-glycolide), polyanhydrides, polyorthoesters, blends and copolymers thereof.
[0273] In some instances, a water insoluble polymer additionally comprises a modified polymer. In some instances, a modified polymer as used herein is a polymer comprising a bioresorbable moiety. In some cases, the term "bioresorbable" refers to a moiety that is capable of being metabolized or broken down and resorbed and/or eliminated through normal excretory routes by the body. In some instances, the metabolites or break down products are non-toxic or substantially non-toxic to the body. In some instances, the bioresorbable moiety is hydrophobic. In other instances, the bioresorbable moiety is hydrophilic. In some cases, the bioresorbable moiety does not render the modified polymer as a whole water soluble.
[0274] Exemplary bioresorbable moieties include, poly(esters), poly(hydroxy acids),
poly(lactones), poly(amides), poly(ester-amides), poly(amino acids), poly(anhydrides), poly(orthoesters), poly(carbonates), poly(phosphazines), poly(phosphoesters), poly(thioesters), polysaccharides and mixtures thereof.
[0275] In some instances, a bioresorbable moiety comprises a resorbable polymer described in U.S. Pat. No. 4,826,945.
[0276] In some instances, a water insoluble polymer comprises components of hydrogels. In some instances, examples of hydrogel components include polyacrylic acids, sodium
carboxymethylcellulose, polyvinyl alcohol, polyvinyl pyrrolidine, gelatin, carrageenan and other polysaccharides, hydroxyethylenemethacrylic acid (HEMA), and derivatives thereof.
[0277] In some instances, a water insoluble polymer comprises a thermoreversible gel. In some cases, examples of thermoreversible gel include pluronics, collagen, gelatin, hyalouronic acid, polysaccharides, polyurethane hydrogel, polyurethane-urea hydrogel or combinations thereof.
[0278] As used herein, a "water-insoluble polymer" includes molecules that are substantially insoluble in water or water-containing environments. In some instances, although certain regions or segments of a polymer are hydrophilic, the polymer molecule, as a whole, does not to any substantial measure dissolve in water.
[0279] In some instances, a FGF-polymer conjugate comprises a FGF-water insoluble polymer conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-biodegradable polymer conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-modified polymer conjugate. In some cases, a FGF-polymer conjugate comprises a FGF -hydrogel conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-thermoreversible gel conjugate. In some cases, a FGF- water insoluble polymer conjugate comprises a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
[0280] In some instances, a polymer modified FGF polypeptide described herein facilitates and/or enables transport of the FGF polypeptide across the BBB. In some instances, a polymer modified FGF polypeptide described herein is further modified with an additional conjugating moiety (e.g., an additional polymer, a protein or its functional fragments thereof, a conjugating peptide, an antibody or its binding fragments thereof, or an antibody mimetic). In some cases, a polymer modified FGF polypeptide described herein with one or more additional conjugating moiety facilitates and/or enables transport of the FGF polypeptide across the BBB.
Lipid
[0281] In some embodiments, a polymer comprises a lipid moiety. Lipid is a broad family of small molecules capable of forming structures such as vesicles, multilamellar/unilamellar liposomes, or membranes in an aqueous environment. In some instances, a lipid comprises a fatty acid. In some cases, a lipid comprises a short chain fatty acid (less than C6), a medium chain fatty acid (C6-C14), or a long chain fatty acid (C15 or longer). In some cases, a fatty acid comprises a saturated fatty acid or an unsaturated fatty acid.
[0282] In some instances, a lipid comprises a saturated fatty acid. Exemplary saturated fatty acid comprises propanoic acid, butanoic acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tridecanoic acid, tetradecanoic acid, pentadecanoic acid, hexadecanoic acid, heptadecanoic acid, octadecanoic acid, nonadecanoic acid, eicosanoic acid, heneicosanoic acid, docosanoic acid, tricosanoic acid, tetracosanoic acid, pentacosanoic acid, hexacosanoic acid, heptacosanoic acid, octacosanoic acid, nonacosanoic acid, triacontanoic acid, henatriacontanoic acid, dotriacontanoic acid, tritriacontanoic acid,
tetratriacontanoic acid, pentatriacontanoic acid, or hexatriacontanoic acid.
[0283] In some cases, a lipid comprises an unsaturated fatty acid. Exemplary unsaturated fatty acid comprises monounsaturated fatty acid such as palmitoleic acid, vaccenic acid, oleic acid, eicosenoic acid, erucic acid, gadoleic acid, myristoleic acid, or nervonic acid; and polyunsaturated fatty acid such as hexadecatrienoic acid, alpha-linolenic acid, stearidonic acid, eicosatrienoic acid, eicosatetraenoic acid, eicosapentaenoic acid, heneicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, tetracosapentaenoic acid, tetracosahexaenoic acid, linoleic acid, gamma-linolenic acid, eicosadienoic acid, dihomo-gamma-linolenic acid, arachidonic acid, docosadienoic acid, adrenic acid,
docosapentaenoic acid, tetracosatetraenoic acid, tetracosapentaenoic acid, mead acid, rumenic acid, alpha-calendic acid, beta-calendic acid, jacaric acid, alpha-eleostearic acid, beta-eleostearic acid, catalpic acid, punicic acid, rumelenic acid, alpha-parinaric acid, beta-parinaric acid, bosseopentaenoic acid, pinolenic acid, or podocarpic acid.
[0284] In some instances, a lipid as described herein comprises components of a liposome, and includes, e.g., phospholipids and/or sterol lipids. In some instances, examples of phospholipids include glycerophospholipids such as glycerophosphocholines, glycerophosphoethanolamines, glycerophosphoserines, glycerophosphoglycerols, glycerophosphoglycerophosphate,
glycerophosphoinositols, glycerophosphoinositol monophosphates, glycerophosphoinositol bisphosphates, glycerophosphoinositol trisphosphates, glycerophosphates, glyceropyrophosphates, glycerophosphoglycerophosphoglycerols, CDP-glycerols, glycosylglycerophospholipids, glycerophosphoinositolglycans, glycerophosphonocholines, glycerophosphonoethanolamines, di- glycerol tetraether phospholipids (caldarchaeols), glycerol-nonitol tetraether phospholipids, or oxidized glycerophospholipids; phosphatidic acid (PA); phosphatidylethanolamie (PE);
phosphatidylcholine (PC); phosphatidylserine (PS); phosphatidylinositol (PI); phosphatidylinositol phosphate (PIP); or phosphosphingolipids such as ceramide phosphorylcholine (SPH), ceramide phosphorylethanolamine (Cer-PE), or ceramide phosphoryllipid. In some instances, examples of sterol lipid include cholesterol, ergosterol, C24-propyl sterols, or stanol. [0285] In some instances, a FGF-polymer conjugate comprises a FGF-lipid conjugate. In some cases, a FGF-polymer conjugate comprises a FGF-fatty acid conjugate. In some cases, the lipid forms a liposome. In some instances, a FGF polypeptide described herein is conjugated or bond to a liposome. In some cases, a FGF polypeptide described herein comprises a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
[0286] In some instances, a lipid modified FGF polypeptide described herein facilitates and/or enables transport of the FGF polypeptide across the BBB. In some instances, a lipid modified FGF polypeptide described herein is further modified with an additional conjugating moiety (e.g., an additional lipid, a polymer, a protein or its functional fragments thereof, a conjugating peptide, an antibody or its binding fragments thereof, or an antibody mimetic). In some cases, a lipid modified FGF polypeptide described herein with one or more additional conjugating moiety facilitates and/or enables transport of the FGF polypeptide across the BBB.
Protein and Functional Fragments
[0287] In some instances, a conjugating moiety comprises a protein or its functional fragments thereof. In some instances, a protein or its functional fragments thereof comprises albumin, transthyretin or thyroxine binding globulin. In some instances, a conjugating moiety is albumin. Albumin is a family of water-soluble globular proteins. It is commonly found in blood plasma, comprising about 55-60% of all plasma proteins. Human serum albumin (HSA) is a 585 amino acid residue polypeptide in which the tertiary structure is divided into three domains, domain I (amino acid residues 1-195), domain II (amino acid residues 196-383), and domain III (amino acid residues 384- 585). Each domain further comprises a binding site, which can interact either reversibly or irreversibly with endogenous ligands such as long- and medium-chain fatty acids, bilirubin, or hemin, or exogenous compounds such as heterocyclic or aromatic compounds.
[0288] In some embodiments, a FGF polypeptide described herein is conjugated to a serum albumin. In some cases, a FGF polypeptide described herein is conjugated to human serum albumin (HSA). In some cases, the conjugation is a covalent conjugation. In some instances, the conjugation is genetic linkage of albumin to a FGF polypeptide. In some cases, the conjugation method is referred to as HSAylation. In some cases, a FGF polypeptide described herein comprises a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
[0289] In some instances, a FGF polypeptide described herein is conjugated to a functional fragment of albumin. In some cases, a functional fragment of albumin comprises an albumin with a N-terminal truncation, a C-terminal truncation or a truncation of both termini. In some cases, a functional fragment of albumin comprises domain I, domain II, domain III, or a combination thereof. In other cases, a functional fragment of albumin comprises a polypeptide of at least 70%, 80%, 90%, or 95% sequence identity to the full-length wild type albumin (e.g., HSA). [0290] In some instances, a FGF polypeptide described herein is conjugated to a functional fragment of HSA. In some cases, a FGF polypeptide described herein is conjugated to a functional fragment of HSA comprising N-terminal truncation, C-terminal truncation or a truncation of both termini. In some cases, a FGF polypeptide described herein is conjugated to a functional fragment of HSA comprising domain I, domain II, domain III, or a combination thereof. In some cases, a FGF polypeptide described herein is conjugated to a functional fragment of HSA comprising at least 70%, 80%, 90%, or 95% sequence identity to the full-length wild type HSA. In some cases, a FGF polypeptide described herein comprises a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922- 25974.
[0291] In some instances, a conjugating moiety is transthyretin (TTR). Transthyretin is a transport protein located in the serum and cerebrospinal fluid which transports the thyroid hormone thyroxine (T4) and retinol-binding protein bound to retinol. In some embodiments, a FGF polypeptide described herein is conjugated to transthyretin (via one of its termini or via an internal hinge region) or its functional fragment thereof. In some cases, a functional fragment of transthyretin comprises a N- terminal truncation, a C-terminal truncation or a truncation of both termini. In some cases, a functional fragment of transthyretin comprises a polypeptide of at least 70%, 80%, 90%, or 95% sequence identity to the full-length wild type transthyretin. In some cases, the conjugation is a covalent conjugation. In some instances, the conjugation is genetic linkage of transthyretin to a FGF polypeptide. In some cases, a FGF polypeptide described herein comprises a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
[0292] In some instances, a conjugating moiety is thyroxine binding globulin (TBG). Thyroxine binding globulin is a globular protein that along with transthyretin transports thyroid hormone thyroxine (T4) and 3,4,3 '-triiodothyronine (T3) in the bloodstream. In some embodiments, a FGF polypeptide described herein is conjugated to TBG or its functional fragment thereof. In some cases, a functional fragment of TBG comprises a N-terminal truncation, a C-terminal truncation or a truncation of both termini. In some cases, a functional fragment of TBG comprises a polypeptide of at least 70%, 80%, 90%, or 95% sequence identity to the full-length wild type TBG. In some cases, the conjugation is a covalent conjugation. In some instances, the conjugation is genetic linkage of TBG to a FGF polypeptide. In some cases, a FGF polypeptide described herein comprises a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
[0293] In some instances, a FGF-protein conjugate described herein facilitates and/or enables transport of the FGF polypeptide across the BBB. In some instances, a FGF-protein conjugate described herein is further modified with an additional conjugating moiety (e.g., an additional protein or its functional fragments thereof, a polymer, a lipid, a conjugating peptide, an antibody or its binding fragments thereof, or an antibody mimetic). In some cases, a FGF-protein conjugate described herein with one or more additional conjugating moiety facilitates and/or enables transport of the FGF polypeptide across the BBB.
Peptide Conjugating Moiety
[0294] In some instances, a conjugating moiety comprises a conjugating peptide. In some instances, a conjugating peptide modulates one or more biological properties and/or physicochemical properties of a target molecule of interest (e.g., a FGF polypeptide). In some instances, a conjugating peptide enhances pharmacokinetics, pharmacodynamics, metabolic stability, biodistribution, water solubility, lipophilicity, or tissue targeting capabilities of a target molecule of interest (e.g., a FGF polypeptide).
[0295] In some instances, a conjugating peptide is derived from an endogenous protein and/or is a fragment of an endogenous protein. In other instances, a conjugating peptide is engineered de novo, such as for example, through a selection screen such as a phase or yeast display.
[0296] In some embodiments, modification of a FGF polypeptide described herein by a glycine- rich homoamino acid polymer (HAP) is referred to as HAPylation. HAPylation is the genetic fusion of a nucleic acid encoding a polypeptide of interest with a nucleic acid encoding a glycine-rich homoamino acid polymer (HAP). In some instances, the HAP polymer comprises a (Gly4Ser)n repeat motif (SEQ ID NO: 25919) and sometimes are about 50, 100, 150, 200, 250, 300, or more residues in length. In some instances, a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974 is modified by HAPylation.
[0297] In some embodiments, modification of a FGF polypeptide described herein by a PAS polypeptide is referred to as PASylation. PASylation is the genetic fusion of a nucleic acid encoding a polypeptide of interest with a nucleic acid encoding a PAS polypeptide. A PAS polypeptide is a hydrophilic uncharged polypeptide consisting of Pro, Ala and Ser residues. In some instances, the length of a PAS polypeptide is at least about 100, 200, 300, 400, 500, or 600 amino acids. In some instances, a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974 is modified by PASylation.
[0298] In some embodiments, modification of a FGF polypeptide described herein by a XTEN™ peptide (Amunix Operating Inc.) is referred to as XTENylation. XTENylation is the genetic fusion of a nucleic acid encoding a polypeptide of interest with a nucleic acid encoding a XTEN™ peptide (Amunix Operating Inc.), a long unstructured hydrophilic peptide comprising different percentage of six amino acids: Ala, Glu, Gly, Ser, and Thr. In some instances, a XTEN™ peptide is selected based on properties such as expression, genetic stability, solubility, aggregation resistance, enhanced half- life, increased potency, and/or increased in vitro activity in combination with a polypeptide of interest. In some instances, a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974 is modified by XTENylation. [0299] In some embodiments, modification of a FGF polypeptide described herein by an elastin- like polypeptide (ELP) is referred to as ELPylation. ELPylation is the genetic fusion of a nucleic acid encoding a polypeptide of interest with a nucleic acid encoding an elastin-like polypeptide (ELPs). An ELP comprises a VPGxG repeat motif (SEQ ID NO: 25920) in which x is any amino acid except proline. In some instances, a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974 is modified by ELPylation.
[0300] In some embodiments, a FGF polypeptide described herein is modified by a CTP peptide. A CTP peptide comprises a 31 amino acid residue peptide
FQSSSS*KAPPPS*LPSPS*RLPGPS*DTPILPQ (SEQ ID NO: 25921) in which the S* denotes O- glycosylation sites (OPKO). In some instances, a CTP peptide is genetically fused to a FGF polypeptide. In some instances, a CTP peptide is genetically fused to a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974.
[0301] In some embodiments, a FGF polypeptide described herein is modified by glutamylation. Glutamylation (or polyglutamylation) is a reversible posttranslational modification of glutamate, in which the γ-carboxy group of glutamate forms a peptide-like bond with the amino group of a free glutamate in which the a-carboxy group extends into a polyglutamate chain. In some instances, a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922-25974 is modified by glutamylation.
[0302] In some instances, a FGF-peptide conjugate described herein facilitates and/or enables transport of the FGF polypeptide across the BBB. In some instances, a FGF-peptide conjugate described herein is further modified with an additional conjugating moiety (e.g., an additional conjugating peptide, a polymer, a protein or its functional fragments thereof, an antibody or its binding fragments thereof, or an antibody mimetic). In some cases, a FGF-peptide conjugate described herein with one or more additional conjugating moiety facilitates and/or enables transport of the FGF polypeptide across the BBB.
Antibody and binding fragments
[0303] In some instances, a conjugating moiety comprises an antibody or its binding fragments thereof. In some instances, an antibody or its binding fragments thereof comprise a humanized antibody or binding fragment thereof, murine antibody or binding fragment thereof, chimeric antibody or binding fragment thereof, monoclonal antibody or binding fragment thereof, monovalent Fab', divalent Fab2, F(ab)'3 fragments, single-chain variable fragment (scFv), bis-scFv, (scFv)2, diabody, minibody, nanobody, triabody, tetrabody, humabody, disulfide stabilized Fv protein (dsFv), single- domain antibody (sdAb), Ig NAR, camelid antibody or binding fragment thereof, bispecific antibody or biding fragment thereof, or a chemically modified derivative thereof.
[0304] In some instances, a FGF polypeptide described herein is conjugated to an antibody or its binding fragments thereof. In some instances, a FGF polypeptide described herein is conjugated to an antibody or its binding fragments thereof selected from a humanized antibody or binding fragment thereof, murine antibody or binding fragment thereof, chimeric antibody or binding fragment thereof, monoclonal antibody or binding fragment thereof, monovalent Fab', divalent Fab2, F(ab)'3 fragments, single-chain variable fragment (scFv), bis-scFv, (scFv)2, diabody, minibody, nanobody, triabody, tetrabody, humabody, disulfide stabilized Fv protein (dsFv), single-domain antibody (sdAb), Ig NAR, camelid antibody or binding fragment thereof, bispecific antibody or biding fragment thereof, or a chemically modified derivative thereof. In some instances, a FGF polypeptide selected from SEQ ID NOs: 3-25917 and 25922-25974 is conjugated to an antibody or its binding fragments thereof selected from a humanized antibody or binding fragment thereof, murine antibody or binding fragment thereof, chimeric antibody or binding fragment thereof, monoclonal antibody or binding fragment thereof, monovalent Fab', divalent Fab2, F(ab)'3 fragments, single-chain variable fragment (scFv), bis-scFv, (scFv)2, diabody, minibody, nanobody, triabody, tetrabody, humabody, disulfide stabilized Fv protein (dsFv), single-domain antibody (sdAb), Ig NAR, camelid antibody or binding fragment thereof, bispecific antibody or biding fragment thereof, or a chemically modified derivative thereof. In some cases, the conjugation is a covalent conjugation. In some instances, the conjugation is genetic linkage of an antibody or its binding fragments thereof to a FGF polypeptide.
[0305] In some instances, an antibody or its binding fragment thereof comprise an antibody or its binding fragments thereof that recognizes transferrin receptor (TfR), insulin receptor (IR), leptin receptor, or lipoprotein-related protein-1 and -2 receptors. In some cases, a FGF polypeptide described herein is conjugated to an antibody or its binding fragments thereof that recognizes transferrin receptor (TfR), insulin receptor (IR), leptin receptor, or lipoprotein-related protein-1 and -2 receptors. In some cases, a FGF polypeptide described herein is conjugated to an antibody or its binding fragments thereof that recognizes transferrin receptor (TfR). In some cases, a FGF polypeptide described herein is conjugated to an antibody or its binding fragments thereof that recognizes insulin receptor (IR). In some cases, a FGF polypeptide described herein is conjugated to an antibody or its binding fragments thereof that recognizes leptin receptor. In some cases, a FGF polypeptide described herein is conjugated to an antibody or its binding fragments thereof that recognizes one or more lipoprotein-related protein-1 and -2 receptors.
[0306] In some embodiments, a FGF polypeptide described herein is conjugated to a transferrin receptor (TfR) interacting antibody or its binding fragments thereof. Exemplary transferrin receptor (TfR) interacting antibodies or their binding fragments thereof include, but are not limited to, 0X26, 8D3, and R17-217. In some embodiments, a FGF polypeptide described herein is conjugated to 0X26. In some embodiments, a FGF polypeptide described herein is conjugated to 8D3. In some embodiments, a FGF polypeptide described herein is conjugated to R17-217. In some instances, a FGF polypeptide described herein is conjugated to a transferrin receptor (TfR) interacting antibody described in Weidler, et al, "The Blood-brain barrier challenge for the treatment of brain cancer, secondary brain metastases, and neurological disease," Cancer Genomics & Proteomics 12: 167-178 (2015).
[0307] In some embodiments, a FGF polypeptide described herein is conjugated to a cell-division cycle 50A interacting antibody or its binding fragments thereof. Exemplary cell-division cycle 50A interacting antibody or its binding fragments thereof includes FC5. In some embodiments, a FGF polypeptide described herein is conjugated to FC5.
[0308] In some embodiments, a FGF polypeptide described herein is conjugated to a ligand of low- density receptor related protein- 1 and -2, receptor-related proteins. In some instances, the ligand is apolipoprotein E. In some cases, the ligand is melanotransferrin/p97. In some embodiments, a FGF polypeptide described herein is conjugated to apolipoprotein E. In some embodiments, a FGF polypeptide described herein is conjugated to melanotransferrin/p97.
[0309] In some instances, a FGF-antibody conjugate described herein facilitates and/or enables transport of the FGF polypeptide across the BBB. In some instances, a FGF-antibody conjugate described herein is further modified with an additional conjugating moiety (e.g., an additional antibody or its binding fragments thereof, a polymer, a protein or its functional fragments thereof, conjugating peptide, or an antibody mimetic). In some cases, a FGF-antibody conjugate described herein with one or more additional conjugating moiety facilitates and/or enables transport of the FGF polypeptide across the BBB.
Antibody Mimetics
[0310] In some embodiments, a conjugating moiety comprises an antibody mimetic. In some cases, an antibody mimetic is an artificial polypeptide with a molar mass of about 3 to about 20 kDa. In some cases, an antibody mimetic is similar to an antibody in that it recognizes and interacts with an antigen, but is not structurally related to an antibody. In some instances, examples of antibody mimetics include Affibody, Affilin, Affimer, Alphabodies, Avimers, DARPin, or Monobodies.
[0311] In some instances, Affibodies are small proteins that can be engineered to bind to a target protein of interest. The scaffold of Affibody is based on the Z domain (or the IgG binding domain) of protein A and comprises a three-helix bundle structure without disulfide bonds.
[0312] In some cases, Affilin (Scil Proteins GmbH) are small proteins derived from human ubiquitin or from gamma-B crystallin. Eight surface exposed residues from either ubiquitin or gamma-B crystalline are further modified for interaction with a target protein of interest.
[0313] In some instances, Affimer is derived from cystatins, a family of cysteine protease inhibitors. The Affimer structure comprises two peptide loops and an N-terminal sequence which in some cases are randomized to generate a target site of interest. [0314] In some instances, Alphabodies (or cell-penetrating alphabodies or CPAB), are small proteins of about 10 kDa in size. Alphabodies comprise coiled-coil structures and the sequences are modified, e.g., by phase display, to generate target site of interest.
[0315] In some cases, Avimers comprise two or more peptide sequences of about 30 to about 35 amino acid residues in length. In some cases, each peptide sequence interacts with an epitope of a target protein of interest, and the combination of peptides to different epitopes of a target protein of interest increases affinity to the target protein.
[0316] In some instances, DARPins (designed Ankyrin repeat proteins) are derived from Ankyrin proteins. In some cases, DARPins comprise of about three or more repeat motifs and the total molecule mass of DARPins is about 14 to about 18 kDa.
[0317] In some cases, Monobodies are derived from fibronectin type III domain (FN3).
Monobodies comprise seven beta sheets forming a beta-sandwich structure with three exposed lops which mimics the three CDRs of an antibody.
[0318] In some embodiments, a FGF polypeptide described herein is conjugated to an antibody mimetic. In some cases, a FGF polypeptide described herein is conjugated to an antibody mimetic selected from Affibody, Affilin, Affimer, Alphabodies, Avimers, DARPin, or Monobodies. In some cases, the conjugation is a covalent conjugation. In some instances, the conjugation is genetic linkage of an antibody or its binding fragments thereof to a FGF polypeptide. In some cases, a FGF polypeptide described herein comprises a FGF polypeptide of SEQ ID NOs: 3-25917 and 25922- 25974.
[0319] In some instances, a FGF-antibody mimetic conjugate described herein facilitates and/or enables transport of the FGF polypeptide across the BBB. In some instances, a FGF-antibody mimetic conjugate described herein is further modified with an additional conjugating moiety (e.g., an additional antibody mimetic, a polymer, a protein or its functional fragments thereof, conjugating peptide, or an antibody or its binding fragments thereof). In some cases, a FGF-antibody mimetic conjugate described herein with one or more additional conjugating moiety facilitates and/or enables transport of the FGF polypeptide across the BBB.
Functional Reactive Groups
[0320] In some embodiments, useful functional reactive groups for conjugating or binding a conjugating moiety to a FGF polypeptide described herein include, for example, zero or higher-order linkers. In some cases, higher-order linkers comprise bifunctional linkers, such as homobifunctional linkers or heterobifunctional linkers. Exemplary homobifuctional linkers include, but are not limited to, Lomant's reagent dithiobis (succinimidylpropionate) DSP, 3'3'-dithiobis(sulfosuccinimidyl proprionate (DTSSP), disuccinimidyl suberate (DSS), bis(sulfosuccinimidyl)suberate (BS), disuccinimidyl tartrate (DST), disulfosuccinimidyl tartrate (sulfo DST), ethylene glycobis(succinimidylsuccinate) (EGS), disuccinimidyl glutarate (DSG), N,N'-disuccinimidyl carbonate (DSC), dimethyl adipimidate (DMA), dimethyl pimelimidate (DMP), dimethyl suberimidate (DMS), dimethyl-3,3'-dithiobispropionimidate (DTBP), l,4-di-3'-(2'- pyridyldithio)propionamido)butane (DPDPB), bismaleimidohexane (BMH), aryl halide-containing compound (DFDNB), such as e.g. l,5-difluoro-2,4-dinitrobenzene or l,3-difluoro-4,6-dinitrobenzene, 4,4'-difluoro-3,3'-dinitrophenylsulfone (DFDNPS), bis-[ -(4-azidosalicylamido)ethyl]disulfide (BASED), formaldehyde, glutaraldehyde, 1,4-butanediol diglycidyl ether, adipic acid dihydrazide, carbohydrazide, o-toluidine, 3,3'-dimethylbenzidine, benzidine, α,α'-ρ-diaminodiphenyl, diiodo-p- xylene sulfonic acid, N,N'-ethylene-bis(iodoacetamide), or N,N'-hexamethylene-bis(iodoacetamide).
[0321] In some embodiments, the bifunctional linker comprises a heterobifunctional linker.
Exemplary heterobifunctional linker include, but are not limited to, amine-reactive and sulfhydryl cross-linkers such as N-succinimidyl 3-(2-pyridyldithio)propionate (sPDP), long -chain N- succinimidyl 3-(2-pyridyldithio)propionate (LC-sPDP), water-soluble-long-chain N-succinimidyl 3- (2-pyridyldithio) propionate (sulfo-LC-sPDP), succinimidyloxycarbonyl-a-methyl-a-(2- pyridyldithio)toluene (sMPT), sulfosuccinimidyl-6-[a-methyl-a-(2-pyridyldithio)toluamido]hexanoate (sulfo-LC-sMPT), succinimidyl-4-(N-maleimidomethyl)cyclohexane- 1 -carboxylate (sMCC), sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane- 1 -carboxylate (sulfo-sMCC), m- maleimidobenzoyl-N-hydroxysuccinimide ester (MBs), m-maleimidobenzoyl-N- hydroxysulfosuccinimide ester (sulfo-MBs), N-succinimidyl(4-iodoacteyl)aminobenzoate (sIAB), sulfosuccinimidyl(4-iodoacteyl)aminobenzoate (sulfo-sIAB), succinimidyl-4-(p- maleimidophenyl)butyrate (sMPB), sulfosuccinimidyl-4-(p-maleimidophenyl)butyrate (sulfo-sMPB), N-(Y-maleimidobutyryloxy)succinimide ester (GMBs), N-(Y-maleimidobutyryloxy)sulfosuccinimide ester (sulfo-GMBs), succinimidyl 6-((iodoacetyl)amino)hexanoate (sIAX), succinimidyl 6-[6- (((iodoacetyl)amino)hexanoyl)amino]hexanoate (sIAXX), succinimidyl 4- (((iodoacetyl)amino)methyl)cyclohexane-l -carboxylate (sIAC), succinimidyl 6-((((4- iodoacetyl)amino)methyl)cyclohexane-l-carbonyl)amino) hexanoate (sIACX), p-nitrophenyl iodoacetate (NPIA), carbonyl-reactive and sulfhydryl-reactive cross-linkers such as 4-(4-N- maleimidophenyl)butyric acid hydrazide (MPBH), 4-(N-maleimidomethyl)cyclohexane-l-carboxyl- hydrazide-8 (M2C2H), 3-(2-pyridyldithio)propionyl hydrazide (PDPH), amine-reactive and photoreactive cross-linkers such as N-hydroxysuccinimidyl-4-azidosalicylic acid (NHs-AsA), N- hydroxysulfosuccinimidyl-4-azidosalicylic acid (sulfo-NHs-AsA), sulfosuccinimidyl-(4- azidosalicylamido)hexanoate (sulfo-NHs-LC-AsA), sulfosuccinimidyl-2-(p-azidosalicylamido)ethyl- l,3'-dithiopropionate (sAsD), N-hydroxysuccinimidyl-4-azidobenzoate (HsAB), N- hydroxysulfosuccinimidyl-4-azidobenzoate (sulfo-HsAB), N-succinimidyl-6-(4'-azido-2'- nitrophenylamino)hexanoate (sANPAH), sulfosuccinimidyl-6-(4'-azido-2'- nitrophenylamino)hexanoate (sulfo-sANPAH), N-5-azido-2-nitrobenzoyloxysuccinimide (ANB- NOs), sulfosuccinimidyl-2-(m-azido-o-nitrobenzamido)-ethyl-l,3'-dithiopropionate (sAND), N- succinimidyl-4(4-azidophenyl) 1 ,3 '-dithiopropionate (sADP), N-sulfosuccinimidyl(4-azidophenyl)- l,3'-dithiopropionate (sulfo-sADP), sulfosuccinimidyl 4-(p-azidophenyl)butyrate (sulfo-sAPB), sulfosuccinimidyl 2-(7-azido-4-methylcoumarin-3-acetamide)ethyl-l,3'-dithiopropionate (sAED), sulfosuccinimidyl 7-azido-4-methylcoumain-3-acetate (sulfo-sAMCA), p-nitrophenyl diazopyruvate (pNPDP), p-nitrophenyl-2-diazo-3,3,3-trifluoropropionate (PNP-DTP), sulfhydryl-reactive and photoreactive cross-linkers such asl-(p-Azidosalicylamido)-4-(iodoacetamido)butane (AsIB), N-[4- (p-azidosalicylamido)butyl]-3'-(2'-pyridyldithio)propionamide (APDP), benzophenone-4- iodoacetamide, benzophenone-4-maleimide carbonyl-reactive and photoreactive cross-linkers such as p-azidobenzoyl hydrazide (ABH), carboxylate-reactive and photoreactive cross-linkers such as 4-(p- azidosalicylamido)butylamine (AsBA), and arginine-reactive and photoreactive cross-linkers such as p-azidophenyl glyoxal (APG).
[0322] In some instances, the reactive functional group comprises a nucleophilic group that is reactive to an electrophilic group present on a binding moiety (e.g., on a conjugating moiety or on a FGF polypeptide). Exemplary electrophilic groups include carbonyl groups— such as aldehyde, ketone, carboxylic acid, ester, amide, enone, acyl halide or acid anhydride. In some embodiments, the reactive functional group is aldehyde. Exemplary nucleophilic groups include hydrazide, oxime, amino, hydrazine, thiosemicarbazone, hydrazine carboxylate, and arylhydrazide.
Pharmaceutical Compositions and Formulations
[0323] In some embodiments, the pharmaceutical composition and formulations described herein are administered to a subject by multiple administration routes, including but not limited to, parenteral (e.g., intravenous, subcutaneous, intramuscular), intracerebral, oral, intranasal, buccal, rectal, or transdermal administration routes. In some instances, the pharmaceutical composition describe herein is formulated for parenteral (e.g., intravenous, subcutaneous, intramuscular) administration. In some instances, the pharmaceutical composition describe herein is formulated for intracerebral (e.g., intrathecal or intraventricular) administration. In other instances, the pharmaceutical composition describe herein is formulated for oral administration. In additional instances, the pharmaceutical composition describe herein is formulated for intranasal administration.
[0324] In some embodiments, the formulation comprises a drug delivery particle. In some cases, a FGF mutant is chemically modified in order to be linked to a drug delivery particle and such chemical modification includes biotinylation, cysteine or lysine conjugation, and the like. In some
embodiments, a FGF mutant is encapsulated within a drug delivery particle. In some instances, the drug delivery particles include liposomes, nanoparticles, micelles, nanoemulsions, microparticles, or microspheres and are comprised of materials including: lipids such as, but not limited to, cholesterol, l,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), l,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), l,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), or hydrogenated soy L-a- phosphatidylcholine (HSPC); synthetic polymers such as, but not limited to, poly(a-hydroxy acids), poly(orthoesters), poly(anhydrides), poly(amino acids), dextrin, poly(glycoside) (PGA), poly(L- lactide) (PLA) and their copolymers poly(D,L-lactic-co-glycolic) acid (PLGA); naturally occurring polymers such as, but not limited to, silk, keratin, collagen, denatured collagen, atelocollagen, gelatin, fibrinogen, fibrin, laminin, elastin, chitosan, hyaluronic acid, starch, carrageenan, cellulose, alginate, perlecan (PLC), heparin or heparin sulfate, chondroitin sulfate, fibronectin, and combinations thereof; polyvinylpyrrollidone (PVP); 2-hydroxyethyl methacrylate (HEMA), or polyamidoamine dendrimers; and nanoparticles such as: paramagnetic nanoparticles, superparamagnetic nanoparticles, metal nanoparticles, fullerene-like materials, inorganic nanotubes, dendrimers (such as with covalently attached metal chelates), nanofibers, nanohorns, nano-onions, nanorods, nanoropes and quantum dots.
[0325] In some instances, the drug delivery particle is a nanoparticle. In some instances, a nanoparticle is a metal nanoparticle, e.g., a nanoparticle of scandium, titanium, vanadium, chromium, manganese, iron, cobalt, nickel, copper, zinc, yttrium, zirconium, niobium, molybdenum, ruthenium, rhodium, palladium, silver, cadmium, hafnium, tantalum, tungsten, rhenium, osmium, iridium, platinum, gold, gadolinium, aluminum, gallium, indium, tin, thallium, lead, bismuth, magnesium, calcium, strontium, barium, lithium, sodium, potassium, boron, silicon, phosphorus, germanium, arsenic, antimony, and combinations, alloys or oxides thereof.
[0326] In some instances, a nanoparticle includes a core or a core and a shell, as in a core-shell nanoparticle.
[0327] In some instances, a nanoparticle is further coated with molecules for attachment of functional elements (e.g., with one or more of a FGF mutant described herein). In some instances, a coating comprises chondroitin sulfate, dextran sulfate, carboxymethyl dextran, alginic acid, pectin, carragheenan, fucoidan, agaropectin, porphyran, karaya gum, gellan gum, xanthan gum, hyaluronic acids, glucosamine, galactosamine, chitin (or chitosan), polyglutamic acid, polyaspartic acid, lysozyme, cytochrome C, ribonuclease, trypsinogen, chymotrypsinogen, a-chymotrypsin, polylysine, polyarginine, histone, protamine, ovalbumin or dextrin or cyclodextrin. In some instances, a nanoparticle comprises a graphene-coated nanoparticle.
[0328] In some cases, a nanoparticle has at least one dimension of less than about 500nm, 400nm, 300nm, 200nm, or lOOnm.
[0329] In some instances, the nanoparticle formulation comprises paramagnetic nanoparticles, superparamagnetic nanoparticles, metal nanoparticles, fullerene-like materials, inorganic nanotubes, dendrimers (such as with covalently attached metal chelates), nanofibers, nanohorns, nano-onions, nanorods, nanoropes or quantum dots. In some instances, a FGF mutant described herein is conjugated either directly or indirectly to the nanoparticle. In some instances, at least 1, 5, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more FGF mutants described herein are conjugated either directly or indirectly to a nanoparticle.
- I l l - [0330] In some instances, the drug delivery particles have pH-triggered release of FGF and are comprised of pH sensitive materials such as, but not limited to, sulphamethazine oligomers (SMOs), sulphonamide and methacrylic acid, poly( amino ester), N-(2-(dimethylamino) ethyl)- methacrylamide (DMAEMA), or pH-sensitive lipids such as dioleoylphosphatidylethanolamine (DOPE).
[0331] In some instances, the drug delivery particles have temperature-triggered release of FGF and are comprised of temperature-sensitive materials such as, but not limited to, poly(N- isopropylacrylamide) (PNIPAAm), poly(propylene glycol), poly(propylene fumarate), or
poly (organophosphazenes) .
[0332] In some embodiments, a FGF mutant is encapsulated within a hydrogel or a scaffold. In some instances, the hydrogels or scaffolds are comprised of synthetic polymers such as, but not limited to, poly(a-hydroxy acids), poly(orthoesters), poly(anhydrides), poly(amino acids), dextrin, poly(glycoside) (PGA), poly(L-lactide) (PLA) and their copolymers poly(D,L-lactic-co-glycolic) acid (PLGA); naturally occurring polymers such as, but not limited to, silk, keratin, collagen, denatured collagen, atelocollagen, gelatin, fibrinogen, fibrin, laminin, elastin, chitosan, hyaluronic acid, starch, carrageenan, cellulose, alginate, perlecan (PLC), heparin or heparin sulfate, chondroitin sulfate, fibronectin, and combinations thereof; polyethylene glycol (PEG); nitrocinnamate-PEG; 2- hydroxyethyl methacrylate (HEMA); polyethersulfone (PES); allogeneic demineralized bone matrix; hydroxy apatite; polycaprolactone; or poly(ethylene oxide) (PEO). In some instances, the hydrogels or scaffolds have pH-triggered release of FGF and are comprised of pH sensitive materials such as, but not limited to, sulphamethazine oligomers (SMOs), sulphonamide and methacrylic acid, poly( amino ester), or N-(2-(dimethylamino) ethyl)-methacrylamide (DMAEMA). In some instances, the hydrogels or scaffolds have temperature-triggered release of FGF and are comprised of temperature- sensitive materials such as, but not limited to, poly(N-isopropylacrylamide) (PNIPAAm), poly(propylene glycol), poly(propylene fumarate), or poly(organophosphazenes). In some embodiments, the hydrogels or scaffolds have protease or enzyme-triggered release of FGF and are comprised of catalytic matrix metalloproteinase (MMP) or ligase-cleavable materials.
[0333] In some embodiments, a viral or plasmid vector encoding a FGF mutant is used as part of the pharmaceutical formulation. In some cases, the type of viruses used to derive the viral vectors include adeno-associated virus (AAV), adenoviruses, herpes simplex viruses, retroviruses, or lentiviruses.
[0334] In some embodiments, the pharmaceutical formulations include, but are not limited to, aqueous liquid dispersions, self-emulsifying dispersions, solid solutions, liposomal dispersions, aerosols, solid dosage forms, powders, immediate release formulations, controlled release formulations, fast melt formulations, tablets, capsules, pills, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations (e.g., nanoparticle formulations), and mixed immediate and controlled release formulations.
[0335] In some embodiments, the pharmaceutical formulations include a carrier or carrier materials selected on the basis of compatibility with the composition disclosed herein, and the release profile properties of the desired dosage form. Exemplary carrier materials include, e.g., binders, suspending agents, disintegration agents, filling agents, surfactants, solubilizers, stabilizers, lubricants, wetting agents, diluents, and the like. Pharmaceutically compatible carrier materials include, but are not limited to, acacia, gelatin, colloidal silicon dioxide, calcium glycerophosphate, calcium lactate, maltodextrin, glycerine, magnesium silicate, polyvinylpyrrollidone (PVP), cholesterol, cholesterol esters, sodium caseinate, soy lecithin, taurocholic acid, phosphotidylcholine, sodium chloride, tricalcium phosphate, dipotassium phosphate, cellulose and cellulose conjugates, sugars sodium stearoyl lactylate, carrageenan, monoglyceride, diglyceride, pregelatinized starch, and the like. See, e.g., Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack
Publishing Company, 1995), Hoover, John E., Remington 's Pharmaceutical Sciences, Mack
Publishing Co., Easton, Pennsylvania 1975, Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY., 1980, and Pharmaceutical Dosage .Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkinsl999).
[0336] In some instances, the pharmaceutical formulations further include pH adjusting agents or buffering agents which include acids such as acetic, boric, citric, lactic, phosphoric and hydrochloric acids, bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate and tris-hydroxymethylaminomethane, and buffers such as citrate/dextrose, sodium bicarbonate and ammonium chloride. Such acids, bases and buffers are included in an amount required to maintain pH of the composition in an acceptable range.
[0337] In some instances, the pharmaceutical formulation includes one or more salts in an amount required to bring osmolality of the composition into an acceptable range. Such salts include those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfite anions, suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite and ammonium sulfate.
[0338] In some embodiments, the pharmaceutical formulations include, but are not limited to, sugars like trehalose, sucrose, mannitol, maltose, glucose, or salts like potassium phosphate, sodium citrate, ammonium sulfate and/or other agents such as heparin to increase the solubility and in vivo stability of polypeptides.
[0339] In some instances, the pharmaceutical formulations further include diluent which are used to stabilize compounds because they can provide a more stable environment. Salts dissolved in buffered solutions (which also can provide pH control or maintenance) are utilized as diluents in the art, including, but not limited to a phosphate buffered saline solution. In certain instances, diluents increase bulk of the composition to facilitate compression or create sufficient bulk for homogenous blend for capsule filling. Such compounds can include e.g., lactose, starch, mannitol, sorbitol, dextrose, microcrystalline cellulose such as Avicel®, dibasic calcium phosphate, dicalcium phosphate dihydrate, tricalcium phosphate, calcium phosphate, anhydrous lactose, spray-dried lactose, pregelatinized starch, compressible sugar, such as Di-Pac® (Amstar), mannitol,
hydroxypropylmethylcellulose, hydroxypropylmethylcellulose acetate stearate, sucrose-based diluents, confectioner's sugar, monobasic calcium sulfate monohydrate, calcium sulfate dihydrate, calcium lactate trihydrate, dextrates, hydrolyzed cereal solids, amylose, powdered cellulose, calcium carbonate, glycine, kaolin, mannitol, sodium chloride, inositol, bentonite, and the like.
[0340] In some cases, the pharmaceutical formulations include disintegration agents or disintegrants to facilitate the breakup or disintegration of a substance. The term "disintegrate" include both the dissolution and dispersion of the dosage form when contacted with gastrointestinal fluid. Examples of disintegration agents include a starch, e.g., a natural starch such as corn starch or potato starch, a pregelatinized starch such as National 1551 or Amijel®, or sodium starch glycolate such as Promogel® or Explotab®, a cellulose such as a wood product, methylcrystalline cellulose, e.g., Avicel®, Avicel® PHlOl, Avicel® PHI 02, Avicel® PHI 05, Elcema® PI 00, Emcocel®, Vivacel®, Ming Tia®, and Solka-Floc®, methylcellulose, croscarmellose, or a cross-linked cellulose, such as cross- linked sodium carboxymethylcellulose (Ac-Di-Sol®), cross-linked carboxymethylcellulose, or cross- linked croscarmellose, a cross-linked starch such as sodium starch glycolate, a cross-linked polymer such as crospovidone, a cross-linked polyvinylpyrrolidone, alginate such as alginic acid or a salt of alginic acid such as sodium alginate, a clay such as Veegum® HV (magnesium aluminum silicate), a gum such as agar, guar, locust bean, Karaya, pectin, or tragacanth, sodium starch glycolate, bentonite, a natural sponge, a surfactant, a resin such as a cation-exchange resin, citrus pulp, sodium lauryl sulfate, sodium lauryl sulfate in combination starch, and the like.
[0341] In some instances, the pharmaceutical formulations include filling agents such as lactose, calcium carbonate, calcium phosphate, dibasic calcium phosphate, calcium sulfate, microcrystalline cellulose, cellulose powder, dextrose, dextrates, dextran, starches, pregelatinized starch, sucrose, xylitol, lactitol, mannitol, sorbitol, sodium chloride, polyethylene glycol, and the like.
[0342] Lubricants and glidants are also optionally included in the pharmaceutical formulations described herein for preventing, reducing or inhibiting adhesion or friction of materials. Exemplary lubricants include, e.g., stearic acid, calcium hydroxide, talc, sodium stearyl fumerate, a hydrocarbon such as mineral oil, or hydrogenated vegetable oil such as hydrogenated soybean oil (Sterotex®), higher fatty acids and their alkali-metal and alkaline earth metal salts, such as aluminum, calcium, magnesium, zinc, stearic acid, sodium stearates, glycerol, talc, waxes, Stearowet®, boric acid, sodium benzoate, sodium acetate, sodium chloride, leucine, a polyethylene glycol (e.g., PEG-4000) or a methoxypolyethylene glycol such as Carbowax™, sodium oleate, sodium benzoate, glyceryl behenate, polyethylene glycol, magnesium or sodium lauryl sulfate, colloidal silica such as Syloid™,
Cab-O-Sil®, a starch such as corn starch, silicone oil, a surfactant, and the like.
[0343] Plasticizers include compounds used to soften the microencapsulation material or film coatings to make them less brittle. Suitable plasticizers include, e.g., polyethylene glycols such as
PEG 300, PEG 400, PEG 600, PEG 1450, PEG 3350, and PEG 800, stearic acid, propylene glycol, oleic acid, triethyl cellulose and triacetin. Plasticizers can also function as dispersing agents or wetting agents.
[0344] Solubilizers include compounds such as triacetin, triethylcitrate, ethyl oleate, ethyl caprylate, sodium lauryl sulfate, sodium doccusate, vitamin E TPGS, dimethylacetamide, N- methylpyrrolidone, N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropylmethyl cellulose, hydroxypropyl cyclodextrins, ethanol, n-butanol, isopropyl alcohol, cholesterol, bile salts, polyethylene glycol 200-600, glycofurol, transcutol, propylene glycol, and dimethyl isosorbide and the like.
[0345] Stabilizers include compounds such as any antioxidation agents, buffers, acids, preservatives and the like. Exemplary stabilizers include L-arginine hydrochloride, tromethamine, albumin (human), citric acid, benzyl alcohol, phenol, disodium biphosphate dehydrate, propylene glycol, metacresol or m-cresol, zinc acetate, polysorbate-20 or Tween® 20, or trometamol.
[0346] Suspending agents include compounds such as polyvinylpyrrolidone, e.g.,
polyvinylpyrrolidone K12, polyvinylpyrrolidone K17, polyvinylpyrrolidone K25, or
polyvinylpyrrolidone K30, vinyl pyrrolidone/vinyl acetate copolymer (S630), polyethylene glycol, e.g., the polyethylene glycol can have a molecular weight of about 300 to about 6000, or about 3350 to about 4000, or about 7000 to about 5400, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, hydroxymethylcellulose acetate stearate, polysorbate-80, hydroxyethylcellulose, sodium alginate, gums, such as, e.g., gum tragacanth and gum acacia, guar gum, xanthans, including xanthan gum, sugars, cellulosics, such as, e.g., sodium
carboxymethylcellulose, methylcellulose, sodium carboxymethylcellulose,
hydroxypropylmethylcellulose, hydroxyethylcellulose, polysorbate-80, sodium alginate,
polyethoxylated sorbitan monolaurate, polyethoxylated sorbitan monolaurate, povidone and the like.
[0347] Surfactants include compounds such as sodium lauryl sulfate, sodium docusate, Tween 60 or 80, triacetin, vitamin E TPGS, sorbitan monooleate, polyoxyethylene sorbitan monooleate, polysorbates, polaxomers, bile salts, glyceryl monostearate, copolymers of ethylene oxide and propylene oxide, e.g., Pluronic® (BASF), and the like. Additional surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, e.g., polyoxyethylene (60) hydrogenated castor oil, and polyoxyethylene alkylethers and alkylphenyl ethers, e.g. , octoxynol 10, octoxynol 40. Sometimes, surfactants is included to enhance physical stability or for other purposes. [0348] Viscosity enhancing agents include, e.g., methyl cellulose, xanthan gum, carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose acetate stearate, hydroxypropylmethyl cellulose phthalate, carbomer, polyvinyl alcohol, alginates, acacia, chitosans and combinations thereof.
[0349] Wetting agents include compounds such as oleic acid, glyceryl monostearate, sorbitan monooleate, sorbitan monolaurate, triethanolamine oleate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monolaurate, sodium docusate, sodium oleate, sodium lauryl sulfate, sodium doccusate, triacetin, Tween 80, vitamin E TPGS, ammonium salts and the like.
Therapeutic Regimens
[0350] In some embodiments, the pharmaceutical compositions described herein are administered for therapeutic applications. In some embodiments, the pharmaceutical composition is administered once per day, twice per day, three times per day or more. The pharmaceutical composition is administered daily, every day, every alternate day, five days a week, once a week, every other week, two weeks per month, three weeks per month, once a month, twice a month, three times per month, or more. The pharmaceutical composition is administered for at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 18 months, 2 years, 3 years, or more.
[0351] In the case wherein the patient's status does improve, upon the doctor's discretion the administration of the composition is given continuously, alternatively, the dose of the composition being administered is temporarily reduced or temporarily suspended for a certain length of time (i.e., a "drug holiday"). In some instances, the length of the drug holiday varies between 2 days and 1 year, including by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. The dose reduction during a drug holiday is from 10%- 100%, including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.
[0352] Once improvement of the patient's conditions has occurred, a maintenance dose is administered if necessary. Subsequently, the dosage or the frequency of administration, or both, can be reduced, as a function of the symptoms, to a level at which the improved disease, disorder or condition is retained.
[0353] In some embodiments, the amount of a given agent that correspond to such an amount varies depending upon factors such as the particular compound, the severity of the disease, the identity (e.g., weight) of the subject or host in need of treatment, but nevertheless is routinely determined in a manner known in the art according to the particular circumstances surrounding the case, including, e.g., the specific agent being administered, the route of administration, and the subject or host being treated. In some instances, the desired dose is conveniently presented in a single dose or as divided doses administered simultaneously (or over a short period of time) or at appropriate intervals, for example as two, three, four or more sub-doses per day.
[0354] The foregoing ranges are merely suggestive, as the number of variables in regard to an individual treatment regime is large, and considerable excursions from these recommended values are not uncommon. Such dosages is altered depending on a number of variables, not limited to the activity of the compound used, the disease or condition to be treated, the mode of administration, the requirements of the individual subject, the severity of the disease or condition being treated, and the judgment of the practitioner.
[0355] In some embodiments, toxicity and therapeutic efficacy of such therapeutic regimens are determined by standard pharmaceutical procedures in cell cultures or experimental animals, including, but not limited to, the determination of the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between the toxic and therapeutic effects is the therapeutic index and it is expressed as the ratio between LD50 and ED50. Compounds exhibiting high therapeutic indices are preferred. The data obtained from cell culture assays and animal studies are used in formulating a range of dosage for use in human. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with minimal toxicity. The dosage varies within this range depending upon the dosage form employed and the route of administration utilized.
Kits/Article of Manufacture
[0356] Disclosed herein, in certain embodiments, are kits and articles of manufacture for use with one or more methods and compositions described herein. Such kits include a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. In one embodiment, the containers are formed from a variety of materials such as glass or plastic.
[0357] The articles of manufacture provided herein contain packaging materials. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, bags, containers, bottles, and any packaging material suitable for a selected formulation and intended mode of administration and treatment.
[0358] For example, the container(s) include one or more of the FGF mutants (e.g., FGF1 mutants or FGF2 mutants) disclosed herein, and optionally one or more pharmaceutical excipients described herein to facilitate the delivery of FGF mutants. Such kits further optionally include an identifying description or label or instructions relating to its use in the methods described herein. [0359] A kit typically includes labels listing contents and/or instructions for use, and package inserts with instructions for use. A set of instructions will also typically be included.
[0360] In one embodiment, a label is on or associated with the container. In one embodiment, a label is on a container when letters, numbers or other characters forming the label are attached, molded or etched into the container itself, a label is associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert. In one
embodiment, a label is used to indicate that the contents are to be used for a specific therapeutic application. The label also indicates directions for use of the contents, such as in the methods described herein.
[0361] In certain embodiments, the pharmaceutical compositions are presented in a pack or dispenser device which contains one or more unit dosage forms containing a compound provided herein. The pack, for example, contains metal or plastic foil, such as a blister pack. In one
embodiment, the pack or dispenser device is accompanied by instructions for administration. In one embodiment, the pack or dispenser is also accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of
pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, is the labeling approved by the U.S. Food and Drug Administration for drugs, or the approved product insert. In one embodiment, compositions containing a compound provided herein formulated in a compatible pharmaceutical carrier are also prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.
Certain Terminology
[0362] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs. It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed. In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that, as used in the specification and the appended claims, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise. In this application, the use of "or" means "and/or" unless stated otherwise. Furthermore, use of the term "including" as well as other forms, such as "include", "includes," and "included," is not limiting.
[0363] As used herein, ranges and amounts can be expressed as "about" a particular value or range. About also includes the exact amount. Hence "about 5 μΙ ' means "about 5 μΙ ' and also "5 μυ" Generally, the term "about" includes an amount that would be expected to be within experimental error. [0364] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.
[0365] As used herein, the terms "individual(s)", "subject(s)" and "patient(s)" mean any mammal. In some embodiments, the mammal is a human. In some embodiments, the mammal is a non-human. None of the terms require or are limited to situations characterized by the supervision (e.g. constant or intermittent) of a health care worker (e.g. a doctor, a registered nurse, a nurse practitioner, a physician's assistant, an orderly or a hospice worker).
EXAMPLES
[0366] These examples are provided for illustrative purposes only and not to limit the scope of the claims provided herein.
Example 1
[0367] Table 1 illustrates the full-length and the mature form of FGFl . Tables 2-12 disclose the SEQ ID NOs associated with the FGF polypeptide sequences described herein. Table 2 is directed to FGFl polypeptides comprising different point mutation(s) and/or N-terminal modifications. Tables 3- 12 illustrate different mutational combinations and optimizations.
Table 1.
Figure imgf000121_0001
Table 2.
Figure imgf000121_0002
MP181 13 MP227 55 MP271 99
MP 182 14 MP228 56 MP272 100
MP 183 15 MP229 57 MP273 101
MP 185 16 MP230 58 MP274 102
MP 186 17 MP231 59 MP275 103
MP 187 18 MP232 60 MP276 104
MP188 19 MP233 61 MP277 105
MP 189 20 MP234 62 MP278 106
MP 190 21 MP235 63 MP279 107
MP191 22 MP236 64 MP280 108
MP 192 23 MP237 65 MP281 109
MP 193 24 MP238 66 MP282 110
MP 194 25 MP239 67 MP283 111
MP 195 26 MP240 68 MP284 112
MP 196 27 MP241 69 MP285 113
MP 197 28 MP242 70 MP286 114
MP 198 29 MP243 71 MP287 115
MP 199 30 MP244 72 MP288 116
MP200 31 MP245 73 MP289 117
MP201 32 MP246 74 MP290 118
MP202 33 MP247 75 MP291 119
MP203 34 MP248 76 MP292 120
MP204 35 MP249 77 MP293 25953
MP205 36 MP250 78 MP294 121
MP206 37 MP251 79 MP295 122
MP207 38 MP252 80 MP296 123
MP208 39 MP253 81 MP297 124
MP209 40 MP254 82 MP298 125
MP210 25951 MP255 83 MP299 126
MP211 41 MP256 84 MP300 127
MP213 42 MP257 85 MP301 128
MP214 43 MP258 86
Table 3.
Figure imgf000122_0001
MP633 Delta nt(10), K12V, H21Y, L44F, N95V, H102Y, F108Y, CI 17V 136
MP634 Delta nt(l l), K12V, H21Y, L44F, N95V, H102Y, F108Y, CI 17V 137
MP635 Delta nt( 11), H21Y, L44F, N95V, H102Y, F108Y, CI 17V 138
MP636 Delta nt(l l), K12V, H21Y, L44F, N95V, H102Y, F108Y, CI 17V, R35E 139
MP637 Delta nt(12), K12V, H21Y, L44F, N95V, H102Y, F108Y, CI 17V 140
MP638 Delta nt(13), K12V, H21Y, L44F, N95V, H102Y, F108Y, CI 17V 141
MP639 Delta nt(9), K12V, H21Y, L44F, N95V, H102Y, F108Y, CI 17V, R35E 142
MP640 Delta nt(9), K12V, H21Y, L44F, N95V, H102Y, F108Y, R35E 143
MP641 Delta nt(10), K12V, Q40P, S47I, H93G, N95V 144
MP642 Delta nt(10), K12V, Q40P, S47I, H93G, N95V, R35E 145
MP643 Delta nt( 11), K12V, Q40P, S47I, H93G, N95V 146
MP644 Delta nt(l 1), K12V, Q40P, S47I, H93G, N95V, R35E 147
MP645 K12V, Q40P, S47I, H93G, N95V, R35E 148
MP646 Delta nt(5), G6A, H21Y, L44F, S47I, H102Y, F108Y, R35E 149
MP647 Delta nt(l l), H21Y, L44F, S47I, H102Y, F108Y 150
MP648 Delta nt(l l), H21Y, L44F, S47I, H102Y, F108Y, R35E 151
MP649 Delta nt(5), G6A, H21Y, Q40P, L44F, H102Y, F108Y, R35E 152
MP650 Delta nt(l l), H21Y, Q40P, L44F, H102Y, F108Y 153
MP651 Delta nt(l l), H21Y, Q40P, L44F, H102Y, F108Y, R35E 154
MP652 Delta nt(l l), R35E 155
MP653 Delta nt(l l), R35E, S116R 156
MP654 Delta nt(l l), K12A, L13A 157
MP655 Delta nt(9), R35E 158
MP656 Delta nt(9), R35E, S 116R 159
MP657 Delta nt(9), K10A, P11A 160
MP658 Delta nt(9), K10A, K12A, L13A, PI 1A 161
MP659 Delta nt(9), K12A + L13A 162
MP690 K12V, R35E, R37E, N95V 163
MP691 K12V, R35E, D36K, R37E, N95V 164
MP692 K12V, R35E, D36R, R37E, N95V 165
MP693 Delta nt(l 1), Q40P, S47I, H93G 166
MP694 Q40P, S47I, H93G, R35E 167
MP695 Delta nt(l 1), Q40P, S47I, H93G, R35E 168
MP696 Delta nt(l l), H21Y, L44F, H102Y, F108Y 169
MP697 H21Y, L44F, H102Y, F108Y, R35E 170
MP698 Delta nt(l l), H21Y, L44F, H102Y, F108Y, R35E 171
MP623 N95V 172
MP624 E87V 173
MP699 Delta nt( 11), K12V, H21Y, L44F, N95V, H102Y, F108Y 174
MP700 Delta nt(9), K12V, Q40P, S47I, H93G, N95V, R35E 175
MP660 Delta nt (7) 176
MP661 Delta nt (8) 177
Figure imgf000124_0001
Table 4.
Figure imgf000124_0002
Table 5.
Figure imgf000124_0003
Table 6.
Figure imgf000125_0001
Table 7.
Figure imgf000125_0002
R88Y 347 E53A 369 Q127A 391
R88D 348 Y55A 370 K128E 392
Y15F 349 Y55F 371 L133V 393
L26A 350 K57A 372
*Base mutation includes Delta nt(l 1), K12V, H21Y, L44F, N95V, H102Y, F108Y
Table 8.
Figure imgf000126_0001
Figure imgf000127_0001
R35E R88D 466 E87K L133A 541 E53A R88Y 616
D36A Y15A 467 E87K Q127K 542 E53A R88D 617
D36A D28K 468 E87K R88L 543
Table 9.
Figure imgf000128_0001
E90K 654 Y8V T30A 716 P11AE53A 778
E91A 655 Y8V T34A 717 P11A Y55A 779
N92A 656 Y8V R35L 718 P11A Y55F 780
T96A 657 Y8V R35G 719 P11AK57A 781
K100E 658 Y8V R35I 720 P11AK57E 782
HI 02 A 659 Y8V R35S 721 P11A Q63A 783
K113A 660 Y8V R35F 722 P11AD68A 784
S116G 661 Y8V R35V 723 P11AT78A 785
S116F 662 Y8V R35P 724 P11AP79A 786
Q127A 663 Y8V R35M 725 P11AE81K 787
K128E 664 Y8V R36K 726 P11AE87V 788
L133V 665 Y8V R37A 727 P11AE87Q 789
Y8AP11A 666 Y8V S38 A 728 P11AL89A 790
Y8A Y15F 667 Y8V D39A 729 P11AE90K 791
Y8A L26A 668 Y8V Q43A 730 P11AE91A 792
Y8A P27A 669 Y8V S47A 731 P11AN92A 793
Y8A T30A 670 Y8V A48G 732 P11AT96A 794
Y8A T34A 671 Y8V E53A 733 P11AK100E 795
Y8A R35L 672 Y8V Y55A 734 P11AH102A 796
Y8A R35G 673 Y8V Y55F 735 P11AK113A 797
Y8A R35I 674 Y8V K57A 736 P11AS116G 798
Y8AR35S 675 Y8V K57E 737 P11AS116F 799
Y8A R35F 676 Y8V Q63A 738 P11AQ127A 800
Y8A R35V 677 Y8V D68A 739 P11AK128E 801
Y8A R35P 678 Y8V T78A 740 P11AL133V 802
Y8A R35M 679 Y8V P79A 741
Table 10.
Figure imgf000129_0001
A48G,K128E 816 K113A,Y64A 2145 T78A,Q43K 3474 Q127A,R88L 4803
A48G,L26A 817 K113A,K10E 2146 T78A,S116H 3475 Q127A,R88Y 4804
A48G,L84A 818 K113A,S17A 2147 T78A,S116Q 3476 Q127A,R88D 4805
A48G,L89A 819 K113A,R122A 2148 T78A,K118M 3477 Q127A,S116W 4806
A48G,N18R 820 K113A,H124A 2149 T78A,K118L 3478 Q127A,S116Y 4807
A48G,N92A 821 K113A,R122E 2150 T78A,C117S 3479 Q127A,S116F 4808
A48G,P121A 822 K113A,S47V 2151 T78A,C117Q 3480 Q127A,S116G 4809
A48G,P27A 823 K113A,S17K 2152 T78A,C117T 3481 Q127A,S116N 4810
A48G,P79A 824 K113A,S116K 2153 T78A,C117P 3482 Q127A,Y15A 4811
A48G,Q63A 825 K113A,N95V 2154 T78A,C117A 3483 Q127A,Y15F 4812
A48G,S38A 826 K113A,Q43K 2155 T78A,C117I 3484 Q127A,Y55A 4813
A48G,S47A 827 K113A,S116H 2156 T96A,C117K 3485 Q127A,Y55F 4814
A48G,T30A 828 K113A,S116Q 2157 T96A,C117R 3486 Q127A,Y94V 4815
A48G,T34A 829 K113A,K118M 2158 T96A,D36K 3487 Q127A,Y94A 4816
A48G,T61A 830 K113A,K118L 2159 T96A,D36A 3488 Q127A,E60A 4817
A48G,T78A 831 K113A,C117S 2160 T96A,E87K 3489 Q127A,Q40A 4818
A48G,T96A 832 K113A,C117Q 2161 T96A,E87Q 3490 Q127A,E60K 4819
A48G,C117K 833 K113A,C117T 2162 T96A,E87V 3491 Q127A,Y64A 4820
A48G,C117R 834 K113A,C117P 2163 T96A,E87D 3492 Q127A,K10E 4821
A48G,D36K 835 K113A,C117A 2164 T96A,K105A 3493 Q127A,S17A 4822
A48G,D36A 836 K113A,C117I 2165 T96A,K105E 3494 Q127A,R122A 4823
A48G,E87K 837 K128E,L26A 2166 T96A,K118W 3495 Q127A,H124A 4824
A48G,E87Q 838 K128E,L84A 2167 T96A,K118H 3496 Q127A,R122E 4825
A48G,E87V 839 K128E,L89A 2168 T96A,K118R 3497 Q127A,S47V 4826
A48G,E87D 840 K128E,N18R 2169 T96A,K118S 3498 Q127A,S17K 4827
A48G,K105A 841 K128E,N92A 2170 T96A,K57A 3499 Q127A,S116K 4828
A48G,K105E 842 K128E,P121A 2171 T96A,K57E 3500 Q127A,N95V 4829
A48G,K118
843 K128E,P27A 2172 T96A,L133A 3501 Q127A,Q43K 4830 W
A48G,K118H 844 K128E,P79A 2173 T96A,L133V 3502 Q127A,S116H 4831
A48G,K118R 845 K128E,Q63A 2174 T96A,N95A 3503 Q127A,S116Q 4832
A48G,K118S 846 K128E,S38A 2175 T96A,N95V 3504 Q127A,K118M 4833
A48G,K57A 847 K128E,S47A 2176 T96A,Q127A 3505 Q127A,K118L 4834
A48G,K57E 848 K128E,T30A 2177 T96A,Q127K 3506 Q127A,C117S 4835
A48G,L133A 849 K128E,T34A 2178 T96A,Q43A 3507 Q127A,C117Q 4836
A48G,L133V 850 K128E/T61A 2179 T96A,Q43E 3508 Q127A,C117T 4837
A48G,N95A 851 K128E,T78A 2180 T96A,R35A 3509 Q127A,C117P 4838
A48G,N95V 852 K128E,T96A 2181 T96A,R35E 3510 Q127A,C117A 4839
A48G,Q127A 853 K128E,C117K 2182 T96A,R35F 3511 Q127A,C117I 4840
A48G,Q127K 854 K128E,C117R 2183 T96A,R35G 3512 Q127K,Q43A 4841
A48G,Q43A 855 K128E,D36K 2184 T96A,R35I 3513 Q127K,Q43E 4842
A48G,Q43E 856 K128E,D36A 2185 T96A,R35L 3514 Q127K,R35A 4843
A48G,R35A 857 K128E,E87K 2186 T96A,R35M 3515 Q127K,R35E 4844
A48G,R35E 858 K128E,E87Q 2187 T96A,R35P 3516 Q127K,R35F 4845
A48G,R35F 859 K128E,E87V 2188 T96A,R35S 3517 Q127K,R35G 4846
A48G,R35G 860 K128E,E87D 2189 T96A,R35V 3518 Q127K,R35I 4847
A48G,R35I 861 K128E,K105A 2190 T96A,R37E 3519 Q127K,R35L 4848 A48G,R35L 862 K128E,K105E 2191 T96A,R37A 3520 Q127K,R35M 4849
A48G,R35M 863 K128E,K118W 2192 T96A,R88L 3521 Q127K,R35P 4850
A48G,R35P 864 K128E,K118H 2193 T96A,R88Y 3522 Q127K,R35S 4851
A48G,R35S 865 K128E,K118R 2194 T96A,R88D 3523 Q127K,R35V 4852
A48G,R35V 866 K128E,K118S 2195 T96A,S116W 3524 Q127K,R37E 4853
A48G,R37E 867 K128E,K57A 2196 T96A,S116Y 3525 Q127K,R37A 4854
A48G,R37A 868 K128E,K57E 2197 T96A,S116F 3526 Q127K,R88L 4855
A48G,R88L 869 K128E,L133A 2198 T96A,S116G 3527 Q127K,R88Y 4856
A48G,R88Y 870 K128E,L133V 2199 T96A,S116N 3528 Q127K,R88D 4857
A48G,R88D 871 K128E,N95A 2200 T96A,Y15A 3529 Q127K,S116W 4858
A48G,S116W 872 K128E,N95V 2201 T96A,Y15F 3530 Q127K,S116Y 4859
A48G,S116Y 873 K128E,Q127A 2202 T96A,Y55A 3531 Q127K,S116F 4860
A48G,S116F 874 K128E,Q127K 2203 T96A,Y55F 3532 Q127K,S116G 4861
A48G,S116G 875 K128E,Q43A 2204 T96A,Y94V 3533 Q127K,S116N 4862
A48G,S116N 876 K128E,Q43E 2205 T96A,Y94A 3534 Q127K,Y15A 4863
A48G,Y15A 877 K128E,R35A 2206 T96A,E60A 3535 Q127K,Y15F 4864
A48G,Y15F 878 K128E,R35E 2207 T96A,Q40A 3536 Q127K,Y55A 4865
A48G,Y55A 879 K128E,R35F 2208 T96A,E60K 3537 Q127K,Y55F 4866
A48G,Y55F 880 K128E,R35G 2209 T96A,Y64A 3538 Q127K,Y94V 4867
A48G,Y94V 881 K128E,R35I 2210 T96A,K10E 3539 Q127K,Y94A 4868
A48G,Y94A 882 K128E,R35L 2211 T96A,S17A 3540 Q127K,E60A 4869
A48G,E60A 883 K128E,R35M 2212 T96A,R122A 3541 Q127K,Q40A 4870
A48G,Q40A 884 K128E,R35P 2213 T96A,H124A 3542 Q127K,E60K 4871
A48G,E60K 885 K128E,R35S 2214 T96A,R122E 3543 Q127K,Y64A 4872
A48G,Y64A 886 K128E,R35V 2215 T96A,S47V 3544 Q127K,K10E 4873
A48G,K10E 887 K128E,R37E 2216 T96A,S17K 3545 Q127K,S17A 4874
A48G,S17A 888 K128E,R37A 2217 T96A,S116K 3546 Q127K,R122A 4875
A48G,R122A 889 K128E,R88L 2218 T96A,N95V 3547 Q127K,H124A 4876
A48G,H124A 890 K128E,R88Y 2219 T96A,Q43K 3548 Q127K,R122E 4877
A48G,R122E 891 K128E,R88D 2220 T96A,S116H 3549 Q127K,S47V 4878
A48G,S47V 892 K128E,S116W 2221 T96A,S116Q 3550 Q127K,S17K 4879
A48G,S17K 893 K128E,S116Y 2222 T96A,K118M 3551 Q127K,S116K 4880
A48G,S116K 894 K128E,S116F 2223 T96A,K118L 3552 Q127K,N95V 4881
A48G,N95V 895 K128E,S116G 2224 T96A,C117S 3553 Q127K,Q43K 4882
A48G,Q43K 896 K128E,S116N 2225 T96A,C117Q 3554 Q127K,S116H 4883
A48G,S116H 897 K128E,Y15A 2226 T96A,C117T 3555 Q127K,S116Q 4884
A48G,S116Q 898 K128E,Y15F 2227 T96A,C117P 3556 Q127K,K118M 4885
A48G,K118
899 K128E,Y55A 2228 T96A,C117A 3557 Q127K,K118L 4886 M
A48G,K118L 900 K128E,Y55F 2229 T96A,C117I 3558 Q127K,C117S 4887
A48G,C117S 901 K128E,Y94V 2230 C117K,D36K 3559 Q127K,C117Q 4888
A48G,C117Q 902 K128E,Y94A 2231 C117K,D36A 3560 Q127K,C117T 4889
A48G,C117T 903 K128E,E60A 2232 C117K,E87K 3561 Q127K,C117P 4890
A48G,C117P 904 K128E,Q40A 2233 C117K,E87Q 3562 Q127K,C117A 4891
A48G,C117A 905 K128E,E60K 2234 C117K,E87V 3563 Q127K,C117I 4892
A48G,C117I 906 K128E,Y64A 2235 C117K,E87D 3564 Q43A,R35A 4893
D28K,D36K 907 K128E,K10E 2236 C117K,K105A 3565 Q43A,R35E 4894 D28K,D39A 908 K128E,S17A 2237 C117K,K105E 3566 Q43A,R35F 4895
D28K,D68A 909 K128E,R122A 2238 C117K,K118W 3567 Q43A,R35G 4896
D28K,E53A 910 K128E,H124A 2239 C117K,K118H 3568 Q43A,R35I 4897
D28K,E81K 911 K128E,R122E 2240 C117K,K118R 3569 Q43A,R35L 4898
D28K,E90K 912 K128E,S47V 2241 C117K,K118S 3570 Q43A,R35M 4899
D28K,E91A 913 K128E,S17K 2242 C117K,K57A 3571 Q43A,R35P 4900
D28K,H102A 914 K128E,S116K 2243 C117K,K57E 3572 Q43A,R35S 4901
D28K,H21A 915 K128E,N95V 2244 C117K,L133A 3573 Q43A,R35V 4902
D28K,K100E 916 K128E,Q43K 2245 C117K,L133V 3574 Q43A,R37E 4903
D28K,K101E 917 K128E,S116H 2246 C117K,N95A 3575 Q43A,R37A 4904
D28K,K113A 918 K128E,S116Q 2247 C117K,N95V 3576 Q43A,R88L 4905
D28K,K128E 919 K128E,K118M 2248 C117K,Q127A 3577 Q43A,R88Y 4906
D28K,L26A 920 K128E,K118L 2249 C117K,Q127K 3578 Q43A,R88D 4907
D28K,L84A 921 K128E,C117S 2250 C117K,Q43A 3579 Q43A,S116W 4908
D28K,L89A 922 K128E,C117Q 2251 C117K,Q43E 3580 Q43A,S116Y 4909
D28K,N18R 923 K128E,C117T 2252 C117K,R35A 3581 Q43A,S116F 4910
D28K,N92A 924 K128E,C117P 2253 C117K,R35E 3582 Q43A,S116G 4911
D28K,P121A 925 K128E,C117A 2254 C117K,R35F 3583 Q43A,S116N 4912
D28K,P27A 926 K128E,C117I 2255 C117K,R35G 3584 Q43A,Y15A 4913
D28K,P79A 927 L26A,L84A 2256 C117K,R35I 3585 Q43A,Y15F 4914
D28K,Q63A 928 L26A,L89A 2257 C117K,R35L 3586 Q43A,Y55A 4915
D28K,S38A 929 L26A,N18R 2258 C117K,R35M 3587 Q43A,Y55F 4916
D28K,S47A 930 L26A,N92A 2259 C117K,R35P 3588 Q43A,Y94V 4917
D28K,T30A 931 L26A,P121A 2260 C117K,R35S 3589 Q43A,Y94A 4918
D28K,T34A 932 L26A,P27A 2261 C117K,R35V 3590 Q43A,E60A 4919
D28K,T61A 933 L26A,P79A 2262 C117K,R37E 3591 Q43A,Q40A 4920
D28K,T78A 934 L26A,Q63A 2263 C117K,R37A 3592 Q43A,E60K 4921
D28K,T96A 935 L26A,S38A 2264 C117K,R88L 3593 Q43A,Y64A 4922
D28K,C117K 936 L26A,S47A 2265 C117K,R88Y 3594 Q43A,K10E 4923
D28K,C117R 937 L26A,T30A 2266 C117K,R88D 3595 Q43A,S17A 4924
D28K,D36K 938 L26A,T34A 2267 C117K,S116W 3596 Q43A,R122A 4925
D28K,D36A 939 L26A,T61A 2268 C117K,S116Y 3597 Q43A,H124A 4926
D28K,E87K 940 L26A,T78A 2269 C117K,S116F 3598 Q43A,R122E 4927
D28K,E87Q 941 L26A,T96A 2270 C117K,S116G 3599 Q43A,S47V 4928
D28K,E87V 942 L26A,C117K 2271 C117K,S116N 3600 Q43A,S17K 4929
D28K,E87D 943 L26A,C117R 2272 C117K,Y15A 3601 Q43A,S116K 4930
D28K,K105A 944 L26A,D36K 2273 C117K,Y15F 3602 Q43A,N95V 4931
D28K,K105E 945 L26A,D36A 2274 C117K,Y55A 3603 Q43A,S116H 4932
D28K,K118
946 L26A,E87K 2275 C117K,Y55F 3604 Q43A,S116Q 4933 W
D28K,K118H 947 L26A,E87Q 2276 C117K,Y94V 3605 Q43A,K118M 4934
D28K,K118R 948 L26A,E87V 2277 C117K,Y94A 3606 Q43A,K118L 4935
D28K,K118S 949 L26A,E87D 2278 C117K,E60A 3607 Q43A,C117S 4936
D28K,K57A 950 L26A,K105A 2279 C117K,Q40A 3608 Q43A,C117Q 4937
D28K,K57E 951 L26A,K105E 2280 C117K,E60K 3609 Q43A,C117T 4938
D28K,L133A 952 L26A,K118W 2281 C117K,Y64A 3610 Q43A,C117P 4939
D28K,L133V 953 L26A,K118H 2282 C117K,K10E 3611 Q43A,C117A 4940 D28K,N95A 954 L26A,K118R 2283 C117K,S17A 3612 Q43A,C117I 4941
D28K,N95V 955 L26A,K118S 2284 C117K,R122A 3613 Q43E,R35A 4942
D28K,Q127A 956 L26A,K57A 2285 C117K,H124A 3614 Q43E,R35E 4943
D28K,Q127K 957 L26A,K57E 2286 C117K,R122E 3615 Q43E,R35F 4944
D28K,Q43A 958 L26A,L133A 2287 C117K,S47V 3616 Q43E,R35G 4945
D28K,Q43E 959 L26A,L133V 2288 C117K,S17K 3617 Q43E,R35I 4946
D28K,R35A 960 L26A,N95A 2289 C117K,S116K 3618 Q43E,R35L 4947
D28K,R35E 961 L26A,N95V 2290 C117K,N95V 3619 Q43E,R35M 4948
D28K,R35F 962 L26A,Q127A 2291 C117K,Q43K 3620 Q43E,R35P 4949
D28K,R35G 963 L26A,Q127K 2292 C117K,S116H 3621 Q43E,R35S 4950
D28K,R35I 964 L26A,Q43A 2293 C117K,S116Q 3622 Q43E,R35V 4951
D28K,R35L 965 L26A,Q43E 2294 C117K,K118M 3623 Q43E,R37E 4952
D28K,R35M 966 L26A,R35A 2295 C117K,K118L 3624 Q43E,R37A 4953
D28K,R35P 967 L26A,R35E 2296 C117R,D36K 3625 Q43E,R88L 4954
D28K,R35S 968 L26A,R35F 2297 C117R,D36A 3626 Q43E,R88Y 4955
D28K,R35V 969 L26A,R35G 2298 C117R,E87K 3627 Q43E,R88D 4956
D28K,R37E 970 L26A,R35I 2299 C117R,E87Q 3628 Q43E,S116W 4957
D28K,R37A 971 L26A,R35L 2300 C117R,E87V 3629 Q43E,S116Y 4958
D28K,R88L 972 L26A,R35M 2301 C117R,E87D 3630 Q43E,S116F 4959
D28K,R88Y 973 L26A,R35P 2302 C117R,K105A 3631 Q43E,S116G 4960
D28K,R88D 974 L26A,R35S 2303 C117R,K105E 3632 Q43E,S116N 4961
D28K,S116W 975 L26A,R35V 2304 C117R,K118W 3633 Q43E,Y15A 4962
D28K,S116Y 976 L26A,R37E 2305 C117R,K118H 3634 Q43E,Y15F 4963
D28K,S116F 977 L26A,R37A 2306 C117R,K118R 3635 Q43E,Y55A 4964
D28K,S116G 978 L26A,R88L 2307 C117R,K118S 3636 Q43E,Y55F 4965
D28K,S116N 979 L26A,R88Y 2308 C117R,K57A 3637 Q43E,Y94V 4966
D28K,Y15A 980 L26A,R88D 2309 C117R,K57E 3638 Q43E,Y94A 4967
D28K,Y15F 981 L26A,S116W 2310 C117R,L133A 3639 Q43E,E60A 4968
D28K,Y55A 982 L26A,S116Y 2311 C117R,L133V 3640 Q43E,Q40A 4969
D28K,Y55F 983 L26A,S116F 2312 C117R,N95A 3641 Q43E,E60K 4970
D28K,Y94V 984 L26A,S116G 2313 C117R,N95V 3642 Q43E,Y64A 4971
D28K,Y94A 985 L26A,S116N 2314 C117R,Q127A 3643 Q43E,K10E 4972
D28K,E60A 986 L26A,Y15A 2315 C117R,Q127K 3644 Q43E,S17A 4973
D28K,Q40A 987 L26A,Y15F 2316 C117R,Q43A 3645 Q43E,R122A 4974
D28K,E60K 988 L26A,Y55A 2317 C117R,Q43E 3646 Q43E,H124A 4975
D28K,Y64A 989 L26A,Y55F 2318 C117R,R35A 3647 Q43E,R122E 4976
D28K,K10E 990 L26A,Y94V 2319 C117R,R35E 3648 Q43E,S47V 4977
D28K,S17A 991 L26A,Y94A 2320 C117R,R35F 3649 Q43E,S17K 4978
D28K,R122A 992 L26A,E60A 2321 C117R,R35G 3650 Q43E,S116K 4979
D28K,H124A 993 L26A,Q40A 2322 C117R,R35I 3651 Q43E,N95V 4980
D28K,R122E 994 L26A,E60K 2323 C117R,R35L 3652 Q43E,S116H 4981
D28K,S47V 995 L26A,Y64A 2324 C117R,R35M 3653 Q43E,S116Q 4982
D28K,S17K 996 L26A,K10E 2325 C117R,R35P 3654 Q43E,K118M 4983
D28K,S116K 997 L26A,S17A 2326 C117R,R35S 3655 Q43E,K118L 4984
D28K,N95V 998 L26A,R122A 2327 C117R,R35V 3656 Q43E,C117S 4985
D28K,Q43K 999 L26A,H124A 2328 C117R,R37E 3657 Q43E,C117Q 4986 D28K,S116H 1000 L26A,R122E 2329 C117R,R37A 3658 Q43E,C117T 4987
D28K,S116Q 1001 L26A,S47V 2330 C117R,R88L 3659 Q43E,C117P 4988
D28K,K118
1002 L26A,S17K 2331 C117R,R88Y 3660 Q43E,C117A 4989 M
D28K,K118L 1003 L26A,S116K 2332 C117R,R88D 3661 Q43E,C117I 4990
D28K,C117S 1004 L26A,N95V 2333 C117R,S116W 3662 R35A,R37E 4991
D28K,C117Q 1005 L26A,Q43K 2334 C117R,S116Y 3663 R35A,R37A 4992
D28K,C117T 1006 L26A,S116H 2335 C117R,S116F 3664 R35A,R88L 4993
D28K,C117P 1007 L26A,S116Q 2336 C117R,S116G 3665 R35A,R88Y 4994
D28K,C117A 1008 L26A,K118M 2337 C117R,S116N 3666 R35A,R88D 4995
D28K,C117I 1009 L26A,K118L 2338 C117R,Y15A 3667 R35A,S116W 4996
D36K,D39A 1010 L26A,C117S 2339 C117R,Y15F 3668 R35A,S116Y 4997
D36K,D68A 1011 L26A,C117Q 2340 C117R,Y55A 3669 R35A,S116F 4998
D36K,E53A 1012 L26A,C117T 2341 C117R,Y55F 3670 R35A,S116G 4999
D36K,E81K 1013 L26A,C117P 2342 C117R,Y94V 3671 R35A,S116N 5000
D36K,E90K 1014 L26A,C117A 2343 C117R,Y94A 3672 R35A,Y15A 5001
D36K,E91A 1015 L26A,C117I 2344 C117R,E60A 3673 R35A,Y15F 5002
D36K,H102A 1016 L84A,L89A 2345 C117R,Q40A 3674 R35A,Y55A 5003
D36K,H21A 1017 L84A,N18R 2346 C117R,E60K 3675 R35A,Y55F 5004
D36K,K100E 1018 L84A,N92A 2347 C117R,Y64A 3676 R35A,Y94V 5005
D36K,K101E 1019 L84A,P121A 2348 C117R,K10E 3677 R35A,Y94A 5006
D36K,K113A 1020 L84A,P27A 2349 C117R,S17A 3678 R35A,E60A 5007
D36K,K128E 1021 L84A,P79A 2350 C117R,R122A 3679 R35A,Q40A 5008
D36K,L26A 1022 L84A,Q63A 2351 C117R,H124A 3680 R35A,E60K 5009
D36K,L84A 1023 L84A,S38A 2352 C117R,R122E 3681 R35A,Y64A 5010
D36K,L89A 1024 L84A,S47A 2353 C117R,S47V 3682 R35A,K10E 5011
D36K,N18R 1025 L84A,T30A 2354 C117R,S17K 3683 R35A,S17A 5012
D36K,N92A 1026 L84A,T34A 2355 C117R,S116K 3684 R35A,R122A 5013
D36K,P121A 1027 L84A,T61A 2356 C117R,N95V 3685 R35A,H124A 5014
D36K,P27A 1028 L84A,T78A 2357 C117R,Q43K 3686 R35A,R122E 5015
D36K,P79A 1029 L84A,T96A 2358 C117R,S116H 3687 R35A,S47V 5016
D36K,Q63A 1030 L84A,C117K 2359 C117R,S116Q 3688 R35A,S17K 5017
D36K,S38A 1031 L84A,C117R 2360 C117R,K118M 3689 R35A,S116K 5018
D36K,S47A 1032 L84A,D36K 2361 C117R,K118L 3690 R35A,N95V 5019
D36K,T30A 1033 L84A,D36A 2362 D36K,E87K 3691 R35A,Q43K 5020
D36K,T34A 1034 L84A,E87K 2363 D36K,E87Q 3692 R35A,S116H 5021
D36K,T61A 1035 L84A,E87Q 2364 D36K,E87V 3693 R35A,S116Q 5022
D36K,T78A 1036 L84A,E87V 2365 D36K,E87D 3694 R35A,K118M 5023
D36K,T96A 1037 L84A,E87D 2366 D36K,K105A 3695 R35A,K118L 5024
D36K,C117K 1038 L84A,K105A 2367 D36K,K105E 3696 R35A,C117S 5025
D36K,C117R 1039 L84A,K105E 2368 D36K,K118W 3697 R35A,C117Q 5026
D36K,E87K 1040 L84A,K118W 2369 D36K,K118H 3698 R35A,C117T 5027
D36K,E87Q 1041 L84A,K118H 2370 D36K,K118R 3699 R35A,C117P 5028
D36K,E87V 1042 L84A,K118R 2371 D36K,K118S 3700 R35A,C117A 5029
D36K,E87D 1043 L84A,K118S 2372 D36K,K57A 3701 R35A,C117I 5030
D36K,K105A 1044 L84A,K57A 2373 D36K,K57E 3702 R35E,R37E 5031
D36K,K105E 1045 L84A,K57E 2374 D36K,L133A 3703 R35E,R37A 5032 D36K,K118
1046 L84A,L133A 2375 D36K,L133V 3704 R35E,R88L 5033 W
D36K,K118H 1047 L84A,L133V 2376 D36K,N95A 3705 R35E,R88Y 5034
D36K,K118R 1048 L84A,N95A 2377 D36K,N95V 3706 R35E,R88D 5035
D36K,K118S 1049 L84A,N95V 2378 D36K,Q127A 3707 R35E,S116W 5036
D36K,K57A 1050 L84A,Q127A 2379 D36K,Q127K 3708 R35E,S116Y 5037
D36K,K57E 1051 L84A,Q127K 2380 D36K,Q43A 3709 R35E,S116F 5038
D36K,L133A 1052 L84A,Q43A 2381 D36K,Q43E 3710 R35E,S116G 5039
D36K,L133V 1053 L84A,Q43E 2382 D36K,R35A 3711 R35E,S116N 5040
D36K,N95A 1054 L84A,R35A 2383 D36K,R35E 3712 R35E,Y15A 5041
D36K,N95V 1055 L84A,R35E 2384 D36K,R35F 3713 R35E,Y15F 5042
D36K,Q127A 1056 L84A,R35F 2385 D36K,R35G 3714 R35E,Y55A 5043
D36K,Q127K 1057 L84A,R35G 2386 D36K,R35I 3715 R35E,Y55F 5044
D36K,Q43A 1058 L84A,R35I 2387 D36K,R35L 3716 R35E,Y94V 5045
D36K,Q43E 1059 L84A,R35L 2388 D36K,R35M 3717 R35E,Y94A 5046
D36K,R35A 1060 L84A,R35M 2389 D36K,R35P 3718 R35E,E60A 5047
D36K,R35E 1061 L84A,R35P 2390 D36K,R35S 3719 R35E,Q40A 5048
D36K,R35F 1062 L84A,R35S 2391 D36K,R35V 3720 R35E,E60K 5049
D36K,R35G 1063 L84A,R35V 2392 D36K,R37E 3721 R35E,Y64A 5050
D36K,R35I 1064 L84A,R37E 2393 D36K,R37A 3722 R35E,K10E 5051
D36K,R35L 1065 L84A,R37A 2394 D36K,R88L 3723 R35E,S17A 5052
D36K,R35M 1066 L84A,R88L 2395 D36K,R88Y 3724 R35E,R122A 5053
D36K,R35P 1067 L84A,R88Y 2396 D36K,R88D 3725 R35E,H124A 5054
D36K,R35S 1068 L84A,R88D 2397 D36K,S116W 3726 R35E,R122E 5055
D36K,R35V 1069 L84A,S116W 2398 D36K,S116Y 3727 R35E,S47V 5056
D36K,R37E 1070 L84A,S116Y 2399 D36K,S116F 3728 R35E,S17K 5057
D36K,R37A 1071 L84A,S116F 2400 D36K,S116G 3729 R35E,S116K 5058
D36K,R88L 1072 L84A,S116G 2401 D36K,S116N 3730 R35E,N95V 5059
D36K,R88Y 1073 L84A,S116N 2402 D36K,Y15A 3731 R35E,Q43K 5060
D36K,R88D 1074 L84A,Y15A 2403 D36K,Y15F 3732 R35E,S116H 5061
D36K,S116W 1075 L84A,Y15F 2404 D36K,Y55A 3733 R35E,S116Q 5062
D36K,S116Y 1076 L84A,Y55A 2405 D36K,Y55F 3734 R35E,K118M 5063
D36K,S116F 1077 L84A,Y55F 2406 D36K,Y94V 3735 R35E,K118L 5064
D36K,S116G 1078 L84A,Y94V 2407 D36K,Y94A 3736 R35E,C117S 5065
D36K,S116N 1079 L84A,Y94A 2408 D36K,E60A 3737 R35E,C117Q 5066
D36K,Y15A 1080 L84A,E60A 2409 D36K,Q40A 3738 R35E,C117T 5067
D36K,Y15F 1081 L84A,Q40A 2410 D36K,E60K 3739 R35E,C117P 5068
D36K,Y55A 1082 L84A,E60K 2411 D36K,Y64A 3740 R35E,C117A 5069
D36K,Y55F 1083 L84A,Y64A 2412 D36K,K10E 3741 R35E,C117I 5070
D36K,Y94V 1084 L84A,K10E 2413 D36K,S17A 3742 R35F,R37E 5071
D36K,Y94A 1085 L84A,S17A 2414 D36K,R122A 3743 R35F,R37A 5072
D36K,E60A 1086 L84A,R122A 2415 D36K,H124A 3744 R35F,R88L 5073
D36K,Q40A 1087 L84A,H124A 2416 D36K,R122E 3745 R35F,R88Y 5074
D36K,E60K 1088 L84A,R122E 2417 D36K,S47V 3746 R35F,R88D 5075
D36K,Y64A 1089 L84A,S47V 2418 D36K,S17K 3747 R35F,S116W 5076
D36K,K10E 1090 L84A,S17K 2419 D36K,S116K 3748 R35F,S116Y 5077
D36K,S17A 1091 L84A,S116K 2420 D36K,N95V 3749 R35F,S116F 5078 D36K,R122A 1092 L84A,N95V 2421 D36K,Q43K 3750 R35F,S116G 5079
D36K,H124A 1093 L84A,Q43K 2422 D36K,S116H 3751 R35F,S116N 5080
D36K,R122E 1094 L84A,S116H 2423 D36K,S116Q 3752 R35F,Y15A 5081
D36K,S47V 1095 L84A,S116Q 2424 D36K,K118M 3753 R35F,Y15F 5082
D36K,S17K 1096 L84A,K118M 2425 D36K,K118L 3754 R35F,Y55A 5083
D36K,S116K 1097 L84A,K118L 2426 D36K,C117S 3755 R35F,Y55F 5084
D36K,N95V 1098 L84A,C117S 2427 D36K,C117Q 3756 R35F,Y94V 5085
D36K,Q43K 1099 L84A,C117Q 2428 D36K,C117T 3757 R35F,Y94A 5086
D36K,S116H 1100 L84A,C117T 2429 D36K,C117P 3758 R35F,E60A 5087
D36K,S116Q 1101 L84A,C117P 2430 D36K,C117A 3759 R35F,Q40A 5088
D36K,K118
1102 L84A,C117A 2431 D36K,C117I 3760 R35F,E60K 5089 M
D36K,K118L 1103 L84A,C117I 2432 D36A,E87K 3761 R35F,Y64A 5090
D36K,C117S 1104 L89A,N18R 2433 D36A,E87Q 3762 R35F,K10E 5091
D36K,C117Q 1105 L89A,N92A 2434 D36A,E87V 3763 R35F,S17A 5092
D36K,C117T 1106 L89A,P121A 2435 D36A,E87D 3764 R35F,R122A 5093
D36K,C117P 1107 L89A,P27A 2436 D36A,K105A 3765 R35F,H124A 5094
D36K,C117A 1108 L89A,P79A 2437 D36A,K105E 3766 R35F,R122E 5095
D36K,C117I 1109 L89A,Q63A 2438 D36A,K118W 3767 R35F,S47V 5096
D39A,D68A 1110 L89A,S38A 2439 D36A,K118H 3768 R35F,S17K 5097
D39A,E53A 1111 L89A,S47A 2440 D36A,K118R 3769 R35F,S116K 5098
D39A,E81K 1112 L89A,T30A 2441 D36A,K118S 3770 R35F,N95V 5099
D39A,E90K 1113 L89A,T34A 2442 D36A,K57A 3771 R35F,Q43K 5100
D39A,E91A 1114 L89A,T61A 2443 D36A,K57E 3772 R35F,S116H 5101
D39A,H102A 1115 L89A,T78A 2444 D36A,L133A 3773 R35F,S116Q 5102
D39A,H21A 1116 L89A,T96A 2445 D36A,L133V 3774 R35F,K118M 5103
D39A,K100E 1117 L89A,C117K 2446 D36A,N95A 3775 R35F,K118L 5104
D39A,K101E 1118 L89A,C117R 2447 D36A,N95V 3776 R35F,C117S 5105
D39A,K113A 1119 L89A,D36K 2448 D36A,Q127A 3777 R35F,C117Q 5106
D39A,K128E 1120 L89A,D36A 2449 D36A,Q127K 3778 R35F,C117T 5107
D39A,L26A 1121 L89A,E87K 2450 D36A,Q43A 3779 R35F,C117P 5108
D39A,L84A 1122 L89A,E87Q 2451 D36A,Q43E 3780 R35F,C117A 5109
D39A,L89A 1123 L89A,E87V 2452 D36A,R35A 3781 R35F,C117I 5110
D39A,N18R 1124 L89A,E87D 2453 D36A,R35E 3782 R35G,R37E 5111
D39A,N92A 1125 L89A,K105A 2454 D36A,R35F 3783 R35G,R37A 5112
D39A,P121A 1126 L89A,K105E 2455 D36A,R35G 3784 R35G,R88L 5113
D39A,P27A 1127 L89A,K118W 2456 D36A,R35I 3785 R35G,R88Y 5114
D39A,P79A 1128 L89A,K118H 2457 D36A,R35L 3786 R35G,R88D 5115
D39A,Q63A 1129 L89A,K118R 2458 D36A,R35M 3787 R35G,S116W 5116
D39A,S38A 1130 L89A,K118S 2459 D36A,R35P 3788 R35G,S116Y 5117
D39A,S47A 1131 L89A,K57A 2460 D36A,R35S 3789 R35G,S116F 5118
D39A,T30A 1132 L89A,K57E 2461 D36A,R35V 3790 R35G,S116G 5119
D39A,T34A 1133 L89A,L133A 2462 D36A,R37E 3791 R35G,S116N 5120
D39A,T61A 1134 L89A,L133V 2463 D36A,R37A 3792 R35G,Y15A 5121
D39A,T78A 1135 L89A,N95A 2464 D36A,R88L 3793 R35G,Y15F 5122
D39A,T96A 1136 L89A,N95V 2465 D36A,R88Y 3794 R35G,Y55A 5123
D39A,C117K 1137 L89A,Q127A 2466 D36A,R88D 3795 R35G,Y55F 5124 D39A,C117R 1138 L89A,Q127K 2467 D36A,S116W 3796 R35G,Y94V 5125
D39A,D36K 1139 L89A,Q43A 2468 D36A,S116Y 3797 R35G,Y94A 5126
D39A,D36A 1140 L89A,Q43E 2469 D36A,S116F 3798 R35G,E60A 5127
D39A,E87K 1141 L89A,R35A 2470 D36A,S116G 3799 R35G,Q40A 5128
D39A,E87Q 1142 L89A,R35E 2471 D36A,S116N 3800 R35G,E60K 5129
D39A,E87V 1143 L89A,R35F 2472 D36A,Y15A 3801 R35G,Y64A 5130
D39A,E87D 1144 L89A,R35G 2473 D36A,Y15F 3802 R35G,K10E 5131
D39A,K105A 1145 L89A,R35I 2474 D36A,Y55A 3803 R35G,S17A 5132
D39A,K105E 1146 L89A,R35L 2475 D36A,Y55F 3804 R35G,R122A 5133
D39A,K118
1147 L89A,R35M 2476 D36A,Y94V 3805 R35G,H124A 5134 W
D39A,K118H 1148 L89A,R35P 2477 D36A,Y94A 3806 R35G,R122E 5135
D39A,K118R 1149 L89A,R35S 2478 D36A,E60A 3807 R35G,S47V 5136
D39A,K118S 1150 L89A,R35V 2479 D36A,Q40A 3808 R35G,S17K 5137
D39A,K57A 1151 L89A,R37E 2480 D36A,E60K 3809 R35G,S116K 5138
D39A,K57E 1152 L89A,R37A 2481 D36A,Y64A 3810 R35G,N95V 5139
D39A,L133A 1153 L89A,R88L 2482 D36A,K10E 3811 R35G,Q43K 5140
D39A,L133V 1154 L89A,R88Y 2483 D36A,S17A 3812 R35G,S116H 5141
D39A,N95A 1155 L89A,R88D 2484 D36A,R122A 3813 R35G,S116Q 5142
D39A,N95V 1156 L89A,S116W 2485 D36A,H124A 3814 R35G,K118M 5143
D39A,Q127A 1157 L89A,S116Y 2486 D36A,R122E 3815 R35G,K118L 5144
D39A,Q127K 1158 L89A,S116F 2487 D36A,S47V 3816 R35G,C117S 5145
D39A,Q43A 1159 L89A,S116G 2488 D36A,S17K 3817 R35G,C117Q 5146
D39A,Q43E 1160 L89A,S116N 2489 D36A,S116K 3818 R35G,C117T 5147
D39A,R35A 1161 L89A,Y15A 2490 D36A,N95V 3819 R35G,C117P 5148
D39A,R35E 1162 L89A,Y15F 2491 D36A,Q43K 3820 R35G,C117A 5149
D39A,R35F 1163 L89A,Y55A 2492 D36A,S116H 3821 R35G,C117I 5150
D39A,R35G 1164 L89A,Y55F 2493 D36A,S116Q 3822 R35I,R37E 5151
D39A,R35I 1165 L89A,Y94V 2494 D36A,K118M 3823 R35I,R37A 5152
D39A,R35L 1166 L89A,Y94A 2495 D36A,K118L 3824 R35I,R88L 5153
D39A,R35M 1167 L89A,E60A 2496 D36A,C117S 3825 R35I,R88Y 5154
D39A,R35P 1168 L89A,Q40A 2497 D36A,C117Q 3826 R35I,R88D 5155
D39A,R35S 1169 L89A,E60K 2498 D36A,C117T 3827 R35I,S116W 5156
D39A,R35V 1170 L89A,Y64A 2499 D36A,C117P 3828 R35I,S116Y 5157
D39A,R37E 1171 L89A,K10E 2500 D36A,C117A 3829 R35I,S116F 5158
D39A,R37A 1172 L89A,S17A 2501 D36A,C117I 3830 R35I,S116G 5159
D39A,R88L 1173 L89A,R122A 2502 E87K,K105A 3831 R35I,S116N 5160
D39A,R88Y 1174 L89A,H124A 2503 E87K,K105E 3832 R35I,Y15A 5161
D39A,R88D 1175 L89A,R122E 2504 E87K,K118W 3833 R35I,Y15F 5162
D39A,S116W 1176 L89A,S47V 2505 E87K,K118H 3834 R35I,Y55A 5163
D39A,S116Y 1177 L89A,S17K 2506 E87K,K118R 3835 R35I,Y55F 5164
D39A,S116F 1178 L89A,S116K 2507 E87K,K118S 3836 R35I,Y94V 5165
D39A,S116G 1179 L89A,N95V 2508 E87K,K57A 3837 R35I,Y94A 5166
D39A,S116N 1180 L89A,Q43K 2509 E87K,K57E 3838 R35I,E60A 5167
D39A,Y15A 1181 L89A,S116H 2510 E87K,L133A 3839 R35I,Q40A 5168
D39A,Y15F 1182 L89A,S116Q 2511 E87K,L133V 3840 R35I,E60K 5169
D39A,Y55A 1183 L89A,K118M 2512 E87K,N95A 3841 R35I,Y64A 5170 D39A,Y55F 1184 L89A,K118L 2513 E87K,N95V 3842 R35I,K10E 5171
D39A,Y94V 1185 L89A,C117S 2514 E87K,Q127A 3843 R35I,S17A 5172
D39A,Y94A 1186 L89A,C117Q 2515 E87K,Q127K 3844 R35I,R122A 5173
D39A,E60A 1187 L89A,C117T 2516 E87K,Q43A 3845 R35I,H124A 5174
D39A,Q40A 1188 L89A,C117P 2517 E87K,Q43E 3846 R35I,R122E 5175
D39A,E60K 1189 L89A,C117A 2518 E87K,R35A 3847 R35I,S47V 5176
D39A,Y64A 1190 L89A,C117I 2519 E87K,R35E 3848 R35I,S17K 5177
D39A,K10E 1191 N18R,N92A 2520 E87K,R35F 3849 R35I,S116K 5178
D39A,S17A 1192 N18R,P121A 2521 E87K,R35G 3850 R35I,N95V 5179
D39A,R122A 1193 N18R,P27A 2522 E87K,R35I 3851 R35I,Q43K 5180
D39A,H124A 1194 N18R,P79A 2523 E87K,R35L 3852 R35I,S116H 5181
D39A,R122E 1195 N18R,Q63A 2524 E87K,R35M 3853 R35I,S116Q 5182
D39A,S47V 1196 N18R,S38A 2525 E87K,R35P 3854 R35I,K118M 5183
D39A,S17K 1197 N18R,S47A 2526 E87K,R35S 3855 R35I,K118L 5184
D39A,S116K 1198 N18R,T30A 2527 E87K,R35V 3856 R35I,C117S 5185
D39A,N95V 1199 N18R,T34A 2528 E87K,R37E 3857 R35I,C117Q 5186
D39A,Q43K 1200 N18R,T61A 2529 E87K,R37A 3858 R35I,C117T 5187
D39A,S116H 1201 N18R,T78A 2530 E87K,R88L 3859 R35I,C117P 5188
D39A,S116Q 1202 N18R,T96A 2531 E87K,R88Y 3860 R35I,C117A 5189
D39A,K118
1203 N18R,C117K 2532 E87K,R88D 3861 R35I,C117I 5190 M
D39A,K118L 1204 N18R,C117R 2533 E87K,S116W 3862 R35L,R37E 5191
D39A,C117S 1205 N18R,D36K 2534 E87K,S116Y 3863 R35L,R37A 5192
D39A,C117Q 1206 N18R,D36A 2535 E87K,S116F 3864 R35L,R88L 5193
D39A,C117T 1207 N18R,E87K 2536 E87K,S116G 3865 R35L,R88Y 5194
D39A,C117P 1208 N18R,E87Q 2537 E87K,S116N 3866 R35L,R88D 5195
D39A,C117A 1209 N18R,E87V 2538 E87K,Y15A 3867 R35L,S116W 5196
D39A,C117I 1210 N18R,E87D 2539 E87K,Y15F 3868 R35L,S116Y 5197
D68A,E53A 1211 N18R,K105A 2540 E87K,Y55A 3869 R35L,S116F 5198
D68A,E81K 1212 N18R,K105E 2541 E87K,Y55F 3870 R35L,S116G 5199
D68A,E90K 1213 N18R,K118W 2542 E87K,Y94V 3871 R35L,S116N 5200
D68A,E91A 1214 N18R,K118H 2543 E87K,Y94A 3872 R35L,Y15A 5201
D68A,H102A 1215 N18R,K118R 2544 E87K,E60A 3873 R35L,Y15F 5202
D68A,H21A 1216 N18R,K118S 2545 E87K,Q40A 3874 R35L,Y55A 5203
D68A,K100E 1217 N18R,K57A 2546 E87K,E60K 3875 R35L,Y55F 5204
D68A,K101E 1218 N18R,K57E 2547 E87K,Y64A 3876 R35L,Y94V 5205
D68A,K113A 1219 N18R,L133A 2548 E87K,K10E 3877 R35L,Y94A 5206
D68A,K128E 1220 N18R,L133V 2549 E87K,S17A 3878 R35L,E60A 5207
D68A,L26A 1221 N18R,N95A 2550 E87K,R122A 3879 R35L,Q40A 5208
D68A,L84A 1222 N18R,N95V 2551 E87K,H124A 3880 R35L,E60K 5209
D68A,L89A 1223 N18R,Q127A 2552 E87K,R122E 3881 R35L,Y64A 5210
D68A,N18R 1224 N18R,Q127K 2553 E87K,S47V 3882 R35L,K10E 5211
D68A,N92A 1225 N18R,Q43A 2554 E87K,S17K 3883 R35L,S17A 5212
D68A,P121A 1226 N18R,Q43E 2555 E87K,S116K 3884 R35L,R122A 5213
D68A,P27A 1227 N18R,R35A 2556 E87K,N95V 3885 R35L,H124A 5214
D68A,P79A 1228 N18R,R35E 2557 E87K,Q43K 3886 R35L,R122E 5215
D68A,Q63A 1229 N18R,R35F 2558 E87K,S116H 3887 R35L,S47V 5216 D68A,S38A 1230 N18R,R35G 2559 E87K,S116Q 3888 R35L,S17K 5217
D68A,S47A 1231 N18R,R35I 2560 E87K,K118M 3889 R35L,S116K 5218
D68A,T30A 1232 N18R,R35L 2561 E87K,K118L 3890 R35L,N95V 5219
D68A,T34A 1233 N18R,R35M 2562 E87K,C117S 3891 R35L,Q43K 5220
D68A,T61A 1234 N18R,R35P 2563 E87K,C117Q 3892 R35L,S116H 5221
D68A,T78A 1235 N18R,R35S 2564 E87K,C117T 3893 R35L,S116Q 5222
D68A,T96A 1236 N18R,R35V 2565 E87K,C117P 3894 R35L,K118M 5223
D68A,C117K 1237 N18R,R37E 2566 E87K,C117A 3895 R35L,K118L 5224
D68A,C117R 1238 N18R,R37A 2567 E87K,C117I 3896 R35L,C117S 5225
D68A,D36K 1239 N18R,R88L 2568 E87Q,K105A 3897 R35L,C117Q 5226
D68A,D36A 1240 N18R,R88Y 2569 E87Q,K105E 3898 R35L,C117T 5227
D68A,E87K 1241 N18R,R88D 2570 E87Q,K118W 3899 R35L,C117P 5228
D68A,E87Q 1242 N18R,S116W 2571 E87Q,K118H 3900 R35L,C117A 5229
D68A,E87V 1243 N18R,S116Y 2572 E87Q,K118R 3901 R35L,C117I 5230
D68A,E87D 1244 N18R,S116F 2573 E87Q,K118S 3902 R35M,R37E 5231
D68A,K105A 1245 N18R,S116G 2574 E87Q,K57A 3903 R35M,R37A 5232
D68A,K105E 1246 N18R,S116N 2575 E87Q,K57E 3904 R35M,R88L 5233
D68A,K118
1247 N18R,Y15A 2576 E87Q,L133A 3905 R35M,R88Y 5234 W
D68A,K118H 1248 N18R,Y15F 2577 E87Q,L133V 3906 R35M,R88D 5235
D68A,K118R 1249 N18R,Y55A 2578 E87Q,N95A 3907 R35M,S116W 5236
D68A,K118S 1250 N18R,Y55F 2579 E87Q,N95V 3908 R35M,S116Y 5237
D68A,K57A 1251 N18R,Y94V 2580 E87Q,Q127A 3909 R35M,S116F 5238
D68A,K57E 1252 N18R,Y94A 2581 E87Q,Q127K 3910 R35M,S116G 5239
D68A,L133A 1253 N18R,E60A 2582 E87Q,Q43A 3911 R35M,S116N 5240
D68A,L133V 1254 N18R,Q40A 2583 E87Q,Q43E 3912 R35M,Y15A 5241
D68A,N95A 1255 N18R,E60K 2584 E87Q,R35A 3913 R35M,Y15F 5242
D68A,N95V 1256 N18R,Y64A 2585 E87Q,R35E 3914 R35M,Y55A 5243
D68A,Q127A 1257 N18R,K10E 2586 E87Q,R35F 3915 R35M,Y55F 5244
D68A,Q127K 1258 N18R,S17A 2587 E87Q,R35G 3916 R35M,Y94V 5245
D68A,Q43A 1259 N18R,R122A 2588 E87Q,R35I 3917 R35M,Y94A 5246
D68A,Q43E 1260 N18R,H124A 2589 E87Q,R35L 3918 R35M,E60A 5247
D68A,R35A 1261 N18R,R122E 2590 E87Q,R35M 3919 R35M,Q40A 5248
D68A,R35E 1262 N18R,S47V 2591 E87Q,R35P 3920 R35M,E60K 5249
D68A,R35F 1263 N18R,S17K 2592 E87Q,R35S 3921 R35M,Y64A 5250
D68A,R35G 1264 N18R,S116K 2593 E87Q,R35V 3922 R35M,K10E 5251
D68A,R35I 1265 N18R,N95V 2594 E87Q,R37E 3923 R35M,S17A 5252
D68A,R35L 1266 N18R,Q43K 2595 E87Q,R37A 3924 R35M,R122A 5253
D68A,R35M 1267 N18R,S116H 2596 E87Q,R88L 3925 R35M,H124A 5254
D68A,R35P 1268 N18R,S116Q 2597 E87Q,R88Y 3926 R35M,R122E 5255
D68A,R35S 1269 N18R,K118M 2598 E87Q,R88D 3927 R35M,S47V 5256
D68A,R35V 1270 N18R,K118L 2599 E87Q,S116W 3928 R35M,S17K 5257
D68A,R37E 1271 N18R,C117S 2600 E87Q,S116Y 3929 R35M,S116K 5258
D68A,R37A 1272 N18R,C117Q 2601 E87Q,S116F 3930 R35M,N95V 5259
D68A,R88L 1273 N18R,C117T 2602 E87Q,S116G 3931 R35M,Q43K 5260
D68A,R88Y 1274 N18R,C117P 2603 E87Q,S116N 3932 R35M,S116H 5261
D68A,R88D 1275 N18R,C117A 2604 E87Q,Y15A 3933 R35M,S116Q 5262 D68A,S116W 1276 N18R,C117I 2605 E87Q,Y15F 3934 R35M,K118M 5263
D68A,S116Y 1277 N92A,P121A 2606 E87Q,Y55A 3935 R35M,K118L 5264
D68A,S116F 1278 N92A,P27A 2607 E87Q,Y55F 3936 R35M,C117S 5265
D68A,S116G 1279 N92A,P79A 2608 E87Q,Y94V 3937 R35M,C117Q 5266
D68A,S116N 1280 N92A,Q63A 2609 E87Q,Y94A 3938 R35M,C117T 5267
D68A,Y15A 1281 N92A,S38A 2610 E87Q,E60A 3939 R35M,C117P 5268
D68A,Y15F 1282 N92A,S47A 2611 E87Q,Q40A 3940 R35M,C117A 5269
D68A,Y55A 1283 N92A,T30A 2612 E87Q,E60K 3941 R35M,C117I 5270
D68A,Y55F 1284 N92A,T34A 2613 E87Q,Y64A 3942 R35P,R37E 5271
D68A,Y94V 1285 N92A,T61A 2614 E87Q,K10E 3943 R35P,R37A 5272
D68A,Y94A 1286 N92A,T78A 2615 E87Q,S17A 3944 R35P,R88L 5273
D68A,E60A 1287 N92A,T96A 2616 E87Q,R122A 3945 R35P,R88Y 5274
D68A,Q40A 1288 N92A,C117K 2617 E87Q,H124A 3946 R35P,R88D 5275
D68A,E60K 1289 N92A,C117R 2618 E87Q,R122E 3947 R35P,S116W 5276
D68A,Y64A 1290 N92A,D36K 2619 E87Q,S47V 3948 R35P,S116Y 5277
D68A,K10E 1291 N92A,D36A 2620 E87Q,S17K 3949 R35P,S116F 5278
D68A,S17A 1292 N92A,E87K 2621 E87Q,S116K 3950 R35P,S116G 5279
D68A,R122A 1293 N92A,E87Q 2622 E87Q,N95V 3951 R35P,S116N 5280
D68A,H124A 1294 N92A,E87V 2623 E87Q,Q43K 3952 R35P,Y15A 5281
D68A,R122E 1295 N92A,E87D 2624 E87Q,S116H 3953 R35P,Y15F 5282
D68A,S47V 1296 N92A,K105A 2625 E87Q,S116Q 3954 R35P,Y55A 5283
D68A,S17K 1297 N92A,K105E 2626 E87Q,K118M 3955 R35P,Y55F 5284
D68A,S116K 1298 N92A,K118W 2627 E87Q,K118L 3956 R35P,Y94V 5285
D68A,N95V 1299 N92A,K118H 2628 E87Q,C117S 3957 R35P,Y94A 5286
D68A,Q43K 1300 N92A,K118R 2629 E87Q,C117Q 3958 R35P,E60A 5287
D68A,S116H 1301 N92A,K118S 2630 E87Q,C117T 3959 R35P,Q40A 5288
D68A,S116Q 1302 N92A,K57A 2631 E87Q,C117P 3960 R35P,E60K 5289
D68A,K118
1303 N92A,K57E 2632 E87Q,C117A 3961 R35P,Y64A 5290 M
D68A,K118L 1304 N92A,L133A 2633 E87Q,C117I 3962 R35P,K10E 5291
D68A,C117S 1305 N92A,L133V 2634 E87V,K105A 3963 R35P,S17A 5292
D68A,C117Q 1306 N92A,N95A 2635 E87V,K105E 3964 R35P,R122A 5293
D68A,C117T 1307 N92A,N95V 2636 E87V,K118W 3965 R35P,H124A 5294
D68A,C117P 1308 N92A,Q127A 2637 E87V,K118H 3966 R35P,R122E 5295
D68A,C117A 1309 N92A,Q127K 2638 E87V,K118R 3967 R35P,S47V 5296
D68A,C117I 1310 N92A,Q43A 2639 E87V,K118S 3968 R35P,S17K 5297
E53A,E81K 1311 N92A,Q43E 2640 E87V,K57A 3969 R35P,S116K 5298
E53A,E90K 1312 N92A,R35A 2641 E87V,K57E 3970 R35P,N95V 5299
E53A,E91A 1313 N92A,R35E 2642 E87V,L133A 3971 R35P,Q43K 5300
E53A,H102A 1314 N92A,R35F 2643 E87V,L133V 3972 R35P,S116H 5301
E53A,H21A 1315 N92A,R35G 2644 E87V,N95A 3973 R35P,S116Q 5302
E53A,K100E 1316 N92A,R35I 2645 E87V,N95V 3974 R35P,K118M 5303
E53A,K101E 1317 N92A,R35L 2646 E87V,Q127A 3975 R35P,K118L 5304
E53A,K113A 1318 N92A,R35M 2647 E87V,Q127K 3976 R35P,C117S 5305
E53A,K128E 1319 N92A,R35P 2648 E87V,Q43A 3977 R35P,C117Q 5306
E53A,L26A 1320 N92A,R35S 2649 E87V,Q43E 3978 R35P,C117T 5307
E53A,L84A 1321 N92A,R35V 2650 E87V,R35A 3979 R35P,C117P 5308 E53A,L89A 1322 N92A,R37E 2651 E87V,R35E 3980 R35P,C117A 5309
E53A,N18R 1323 N92A,R37A 2652 E87V,R35F 3981 R35P,C117I 5310
E53A,N92A 1324 N92A,R88L 2653 E87V,R35G 3982 R35S,R37E 5311
E53A,P121A 1325 N92A,R88Y 2654 E87V,R35I 3983 R35S,R37A 5312
E53A,P27A 1326 N92A,R88D 2655 E87V,R35L 3984 R35S,R88L 5313
E53A,P79A 1327 N92A,S116W 2656 E87V,R35M 3985 R35S,R88Y 5314
E53A,Q63A 1328 N92A,S116Y 2657 E87V,R35P 3986 R35S,R88D 5315
E53A,S38A 1329 N92A,S116F 2658 E87V,R35S 3987 R35S,S116W 5316
E53A,S47A 1330 N92A,S116G 2659 E87V,R35V 3988 R35S,S116Y 5317
E53A,T30A 1331 N92A,S116N 2660 E87V,R37E 3989 R35S,S116F 5318
E53A,T34A 1332 N92A,Y15A 2661 E87V,R37A 3990 R35S,S116G 5319
E53A,T61A 1333 N92A,Y15F 2662 E87V,R88L 3991 R35S,S116N 5320
E53A,T78A 1334 N92A,Y55A 2663 E87V,R88Y 3992 R35S,Y15A 5321
E53A,T96A 1335 N92A,Y55F 2664 E87V,R88D 3993 R35S,Y15F 5322
E53A,C117K 1336 N92A,Y94V 2665 E87V,S116W 3994 R35S,Y55A 5323
E53A,C117R 1337 N92A,Y94A 2666 E87V,S116Y 3995 R35S,Y55F 5324
E53A,D36K 1338 N92A,E60A 2667 E87V,S116F 3996 R35S,Y94V 5325
E53A,D36A 1339 N92A,Q40A 2668 E87V,S116G 3997 R35S,Y94A 5326
E53A,E87K 1340 N92A,E60K 2669 E87V,S116N 3998 R35S,E60A 5327
E53A,E87Q 1341 N92A,Y64A 2670 E87V,Y15A 3999 R35S,Q40A 5328
E53A,E87V 1342 N92A,K10E 2671 E87V,Y15F 4000 R35S,E60K 5329
E53A,E87D 1343 N92A,S17A 2672 E87V,Y55A 4001 R35S,Y64A 5330
E53A,K105A 1344 N92A,R122A 2673 E87V,Y55F 4002 R35S,K10E 5331
E53A,K105E 1345 N92A,H124A 2674 E87V,Y94V 4003 R35S,S17A 5332
E53A,K118W 1346 N92A,R122E 2675 E87V,Y94A 4004 R35S,R122A 5333
E53A,K118H 1347 N92A,S47V 2676 E87V,E60A 4005 R35S,H124A 5334
E53A,K118R 1348 N92A,S17K 2677 E87V,Q40A 4006 R35S,R122E 5335
E53A,K118S 1349 N92A,S116K 2678 E87V,E60K 4007 R35S,S47V 5336
E53A,K57A 1350 N92A,N95V 2679 E87V,Y64A 4008 R35S,S17K 5337
E53A,K57E 1351 N92A,Q43K 2680 E87V,K10E 4009 R35S,S116K 5338
E53A,L133A 1352 N92A,S116H 2681 E87V,S17A 4010 R35S,N95V 5339
E53A,L133V 1353 N92A,S116Q 2682 E87V,R122A 4011 R35S,Q43K 5340
E53A,N95A 1354 N92A,K118M 2683 E87V,H124A 4012 R35S,S116H 5341
E53A,N95V 1355 N92A,K118L 2684 E87V,R122E 4013 R35S,S116Q 5342
E53A,Q127A 1356 N92A,C117S 2685 E87V,S47V 4014 R35S,K118M 5343
E53A,Q127K 1357 N92A,C117Q 2686 E87V,S17K 4015 R35S,K118L 5344
E53A,Q43A 1358 N92A,C117T 2687 E87V,S116K 4016 R35S,C117S 5345
E53A,Q43E 1359 N92A,C117P 2688 E87V,N95V 4017 R35S,C117Q 5346
E53A,R35A 1360 N92A,C117A 2689 E87V,Q43K 4018 R35S,C117T 5347
E53A,R35E 1361 N92A,C117I 2690 E87V,S116H 4019 R35S,C117P 5348
E53A,R35F 1362 P121A,P27A 2691 E87V,S116Q 4020 R35S,C117A 5349
E53A,R35G 1363 P121A,P79A 2692 E87V,K118M 4021 R35S,C117I 5350
E53A,R35I 1364 P121A,Q63A 2693 E87V,K118L 4022 R35V,R37E 5351
E53A,R35L 1365 P121A,S38A 2694 E87V,C117S 4023 R35V,R37A 5352
E53A,R35M 1366 P121A,S47A 2695 E87V,C117Q 4024 R35V,R88L 5353
E53A,R35P 1367 P121A/T30A 2696 E87V,C117T 4025 R35V,R88Y 5354 E53A,R35S 1368 P121A/T34A 2697 E87V,C117P 4026 R35V,R88D 5355
E53A,R35V 1369 P121A/T61A 2698 E87V,C117A 4027 R35V,S116W 5356
E53A,R37E 1370 P121A/T78A 2699 E87V,C117I 4028 R35V,S116Y 5357
E53A,R37A 1371 P121A,T96A 2700 E87D,K105A 4029 R35V,S116F 5358
E53A,R88L 1372 P121A,C117K 2701 E87D,K105E 4030 R35V,S116G 5359
E53A,R88Y 1373 P121A,C117R 2702 E87D,K118W 4031 R35V,S116N 5360
E53A,R88D 1374 P121A,D36K 2703 E87D,K118H 4032 R35V,Y15A 5361
E53A,S116W 1375 P121A,D36A 2704 E87D,K118R 4033 R35V,Y15F 5362
E53A,S116Y 1376 P121A,E87K 2705 E87D,K118S 4034 R35V,Y55A 5363
E53A,S116F 1377 P121A,E87Q 2706 E87D,K57A 4035 R35V,Y55F 5364
E53A,S116G 1378 P121A,E87V 2707 E87D,K57E 4036 R35V,Y94V 5365
E53A,S116N 1379 P121A,E87D 2708 E87D,L133A 4037 R35V,Y94A 5366
E53A,Y15A 1380 P121A,K105A 2709 E87D,L133V 4038 R35V,E60A 5367
E53A,Y15F 1381 P121A,K105E 2710 E87D,N95A 4039 R35V,Q40A 5368
E53A,Y55A 1382 P121A,K118W 2711 E87D,N95V 4040 R35V,E60K 5369
E53A,Y55F 1383 P121A,K118H 2712 E87D,Q127A 4041 R35V,Y64A 5370
E53A,Y94V 1384 P121A,K118R 2713 E87D,Q127K 4042 R35V,K10E 5371
E53A,Y94A 1385 P121A,K118S 2714 E87D,Q43A 4043 R35V,S17A 5372
E53A,E60A 1386 P121A,K57A 2715 E87D,Q43E 4044 R35V,R122A 5373
E53A,Q40A 1387 P121A,K57E 2716 E87D,R35A 4045 R35V,H124A 5374
E53A,E60K 1388 P121A,L133A 2717 E87D,R35E 4046 R35V,R122E 5375
E53A,Y64A 1389 P121A,L133V 2718 E87D,R35F 4047 R35V,S47V 5376
E53A,K10E 1390 P121A,N95A 2719 E87D,R35G 4048 R35V,S17K 5377
E53A,S17A 1391 P121A,N95V 2720 E87D,R35I 4049 R35V,S116K 5378
E53A,R122A 1392 P121A,Q127A 2721 E87D,R35L 4050 R35V,N95V 5379
E53A,H124A 1393 P121A,Q127K 2722 E87D,R35M 4051 R35V,Q43K 5380
E53A,R122E 1394 P121A,Q43A 2723 E87D,R35P 4052 R35V,S116H 5381
E53A,S47V 1395 P121A,Q43E 2724 E87D,R35S 4053 R35V,S116Q 5382
E53A,S17K 1396 P121A,R35A 2725 E87D,R35V 4054 R35V,K118M 5383
E53A,S116K 1397 P121A,R35E 2726 E87D,R37E 4055 R35V,K118L 5384
E53A,N95V 1398 P121A,R35F 2727 E87D,R37A 4056 R35V,C117S 5385
E53A,Q43K 1399 P121A,R35G 2728 E87D,R88L 4057 R35V,C117Q 5386
E53A,S116H 1400 P121A,R35I 2729 E87D,R88Y 4058 R35V,C117T 5387
E53A,S116Q 1401 P121A,R35L 2730 E87D,R88D 4059 R35V,C117P 5388
E53A,K118M 1402 P121A,R35M 2731 E87D,S116W 4060 R35V,C117A 5389
E53A,K118L 1403 P121A,R35P 2732 E87D,S116Y 4061 R35V,C117I 5390
E53A,C117S 1404 P121A,R35S 2733 E87D,S116F 4062 R37E,R88L 5391
E53A,C117Q 1405 P121A,R35V 2734 E87D,S116G 4063 R37E,R88Y 5392
E53A,C117T 1406 P121A,R37E 2735 E87D,S116N 4064 R37E,R88D 5393
E53A,C117P 1407 P121A,R37A 2736 E87D,Y15A 4065 R37E,S116W 5394
E53A,C117A 1408 P121A,R88L 2737 E87D,Y15F 4066 R37E,S116Y 5395
E53A,C117I 1409 P121A,R88Y 2738 E87D,Y55A 4067 R37E,S116F 5396
E81K,E90K 1410 P121A,R88D 2739 E87D,Y55F 4068 R37E,S116G 5397
E81K,E91A 1411 P121A,S116W 2740 E87D,Y94V 4069 R37E,S116N 5398
E81K,H102A 1412 P121A,S116Y 2741 E87D,Y94A 4070 R37E,Y15A 5399
E81K,H21A 1413 P121A,S116F 2742 E87D,E60A 4071 R37E,Y15F 5400 E81K,K100E 1414 P121A,S116G 2743 E87D,Q40A 4072 R37E,Y55A 5401
E81K,K101E 1415 P121A,S116N 2744 E87D,E60K 4073 R37E,Y55F 5402
E81K,K113A 1416 P121A,Y15A 2745 E87D,Y64A 4074 R37E,Y94V 5403
E81K,K128E 1417 P121A,Y15F 2746 E87D,K10E 4075 R37E,Y94A 5404
E81K,L26A 1418 P121A,Y55A 2747 E87D,S17A 4076 R37E,E60A 5405
E81K,L84A 1419 P121A,Y55F 2748 E87D,R122A 4077 R37E,Q40A 5406
E81K,L89A 1420 P121A,Y94V 2749 E87D,H124A 4078 R37E,E60K 5407
E81K,N18R 1421 P121A,Y94A 2750 E87D,R122E 4079 R37E,Y64A 5408
E81K,N92A 1422 P121A,E60A 2751 E87D,S47V 4080 R37E,K10E 5409
E81K,P121A 1423 P121A,Q40A 2752 E87D,S17K 4081 R37E,S17A 5410
E81K,P27A 1424 P121A,E60K 2753 E87D,S116K 4082 R37E,R122A 5411
E81K,P79A 1425 P121A,Y64A 2754 E87D,N95V 4083 R37E,H124A 5412
E81K,Q63A 1426 P121A,K10E 2755 E87D,Q43K 4084 R37E,R122E 5413
E81K,S38A 1427 P121A,S17A 2756 E87D,S116H 4085 R37E,S47V 5414
E81K,S47A 1428 P121A,R122A 2757 E87D,S116Q 4086 R37E,S17K 5415
E81K,T30A 1429 P121A,H124A 2758 E87D,K118M 4087 R37E,S116K 5416
E81K,T34A 1430 P121A,R122E 2759 E87D,K118L 4088 R37E,N95V 5417
Ε81Κ/Γ61Α 1431 P121A,S47V 2760 E87D,C117S 4089 R37E,Q43K 5418
E81K,T78A 1432 P121A,S17K 2761 E87D,C117Q 4090 R37E,S116H 5419
E81K,T96A 1433 P121A,S116K 2762 E87D,C117T 4091 R37E,S116Q 5420
E81K,C117K 1434 P121A,N95V 2763 E87D,C117P 4092 R37E,K118M 5421
E81K,C117R 1435 P121A,Q43K 2764 E87D,C117A 4093 R37E,K118L 5422
E81K,D36K 1436 P121A,S116H 2765 E87D,C117I 4094 R37E,C117S 5423
E81K,D36A 1437 P121A,S116Q 2766 K105A,K118W 4095 R37E,C117Q 5424
E81K,E87K 1438 P121A,K118M 2767 K105A,K118H 4096 R37E,C117T 5425
E81K,E87Q 1439 P121A,K118L 2768 K105A,K118R 4097 R37E,C117P 5426
E81K,E87V 1440 P121A,C117S 2769 K105A,K118S 4098 R37E,C117A 5427
E81K,E87D 1441 P121A,C117Q 2770 K105A,K57A 4099 R37E,C117I 5428
E81K,K105A 1442 P121A,C117T 2771 K105A,K57E 4100 R37A,R88L 5429
E81K,K105E 1443 P121A,C117P 2772 K105A,L133A 4101 R37A,R88Y 5430
E81K,K118W 1444 P121A,C117A 2773 K105A,L133V 4102 R37A,R88D 5431
E81K,K118H 1445 P121A,C117I 2774 K105A,N95A 4103 R37A,S116W 5432
E81K,K118R 1446 P27A,P79A 2775 K105A,N95V 4104 R37A,S116Y 5433
E81K,K118S 1447 P27A,Q63A 2776 K105A,Q127A 4105 R37A,S116F 5434
E81K,K57A 1448 P27A,S38A 2777 K105A,Q127K 4106 R37A,S116G 5435
E81K,K57E 1449 P27A,S47A 2778 K105A,Q43A 4107 R37A,S116N 5436
E81K,L133A 1450 P27A,T30A 2779 K105A,Q43E 4108 R37A,Y15A 5437
E81K,L133V 1451 P27A,T34A 2780 K105A,R35A 4109 R37A,Y15F 5438
E81K,N95A 1452 P27A,T61A 2781 K105A,R35E 4110 R37A,Y55A 5439
E81K,N95V 1453 P27A,T78A 2782 K105A,R35F 4111 R37A,Y55F 5440
E81K,Q127A 1454 P27A,T96A 2783 K105A,R35G 4112 R37A,Y94V 5441
E81K,Q127K 1455 P27A,C117K 2784 K105A,R35I 4113 R37A,Y94A 5442
E81K,Q43A 1456 P27A,C117R 2785 K105A,R35L 4114 R37A,E60A 5443
E81K,Q43E 1457 P27A,D36K 2786 K105A,R35M 4115 R37A,Q40A 5444
E81K,R35A 1458 P27A,D36A 2787 K105A,R35P 4116 R37A,E60K 5445
E81K,R35E 1459 P27A,E87K 2788 K105A,R35S 4117 R37A,Y64A 5446 E81K,R35F 1460 P27A,E87Q 2789 K105A,R35V 4118 R37A,K10E 5447
E81K,R35G 1461 P27A,E87V 2790 K105A,R37E 4119 R37A,S17A 5448
E81K,R35I 1462 P27A,E87D 2791 K105A,R37A 4120 R37A,R122A 5449
E81K,R35L 1463 P27A,K105A 2792 K105A,R88L 4121 R37A,H124A 5450
E81K,R35M 1464 P27A,K105E 2793 K105A,R88Y 4122 R37A,R122E 5451
E81K,R35P 1465 P27A,K118W 2794 K105A,R88D 4123 R37A,S47V 5452
E81K,R35S 1466 P27A,K118H 2795 K105A,S116W 4124 R37A,S17K 5453
E81K,R35V 1467 P27A,K118R 2796 K105A,S116Y 4125 R37A,S116K 5454
E81K,R37E 1468 P27A,K118S 2797 K105A,S116F 4126 R37A,N95V 5455
E81K,R37A 1469 P27A,K57A 2798 K105A,S116G 4127 R37A,Q43K 5456
E81K,R88L 1470 P27A,K57E 2799 K105A,S116N 4128 R37A,S116H 5457
E81K,R88Y 1471 P27A,L133A 2800 K105A,Y15A 4129 R37A,S116Q 5458
E81K,R88D 1472 P27A,L133V 2801 K105A,Y15F 4130 R37A,K118M 5459
E81K,S116W 1473 P27A,N95A 2802 K105A,Y55A 4131 R37A,K118L 5460
E81K,S116Y 1474 P27A,N95V 2803 K105A,Y55F 4132 R37A,C117S 5461
E81K,S116F 1475 P27A,Q127A 2804 K105A,Y94V 4133 R37A,C117Q 5462
E81K,S116G 1476 P27A,Q127K 2805 K105A,Y94A 4134 R37A,C117T 5463
E81K,S116N 1477 P27A,Q43A 2806 K105A,E60A 4135 R37A,C117P 5464
E81K,Y15A 1478 P27A,Q43E 2807 K105A,Q40A 4136 R37A,C117A 5465
E81K,Y15F 1479 P27A,R35A 2808 K105A,E60K 4137 R37A,C117I 5466
E81K,Y55A 1480 P27A,R35E 2809 K105A,Y64A 4138 R88L,S116W 5467
E81K,Y55F 1481 P27A,R35F 2810 K105A,K10E 4139 R88L,S116Y 5468
E81K,Y94V 1482 P27A,R35G 2811 K105A,S17A 4140 R88L,S116F 5469
E81K,Y94A 1483 P27A,R35I 2812 K105A,R122A 4141 R88L,S116G 5470
E81K,E60A 1484 P27A,R35L 2813 K105A,H124A 4142 R88L,S116N 5471
E81K,Q40A 1485 P27A,R35M 2814 K105A,R122E 4143 R88L,Y15A 5472
E81K,E60K 1486 P27A,R35P 2815 K105A,S47V 4144 R88L,Y15F 5473
E81K,Y64A 1487 P27A,R35S 2816 K105A,S17K 4145 R88L,Y55A 5474
E81K,K10E 1488 P27A,R35V 2817 K105A,S116K 4146 R88L,Y55F 5475
E81K,S17A 1489 P27A,R37E 2818 K105A,N95V 4147 R88L,Y94V 5476
E81K,R122A 1490 P27A,R37A 2819 K105A,Q43K 4148 R88L,Y94A 5477
E81K,H124A 1491 P27A,R88L 2820 K105A,S116H 4149 R88L,E60A 5478
E81K,R122E 1492 P27A,R88Y 2821 K105A,S116Q 4150 R88L,Q40A 5479
E81K,S47V 1493 P27A,R88D 2822 K105A,K118M 4151 R88L,E60K 5480
E81K,S17K 1494 P27A,S116W 2823 K105A,K118L 4152 R88L,Y64A 5481
E81K,S116K 1495 P27A,S116Y 2824 K105A,C117S 4153 R88L,K10E 5482
E81K,N95V 1496 P27A,S116F 2825 K105A,C117Q 4154 R88L,S17A 5483
E81K,Q43K 1497 P27A,S116G 2826 K105A,C117T 4155 R88L,R122A 5484
E81K,S116H 1498 P27A,S116N 2827 K105A,C117P 4156 R88L,H124A 5485
E81K,S116Q 1499 P27A,Y15A 2828 K105A,C117A 4157 R88L,R122E 5486
E81K,K118M 1500 P27A,Y15F 2829 K105A,C117I 4158 R88L,S47V 5487
E81K,K118L 1501 P27A,Y55A 2830 K105E,K118W 4159 R88L,S17K 5488
E81K,C117S 1502 P27A,Y55F 2831 K105E,K118H 4160 R88L,S116K 5489
E81K,C117Q 1503 P27A,Y94V 2832 K105E,K118R 4161 R88L,N95V 5490
E81K,C117T 1504 P27A,Y94A 2833 K105E,K118S 4162 R88L,Q43K 5491
E81K,C117P 1505 P27A,E60A 2834 K105E,K57A 4163 R88L,S116H 5492 E81K,C117A 1506 P27A,Q40A 2835 K105E,K57E 4164 R88L,S116Q 5493
E81K,C117I 1507 P27A,E60K 2836 K105E,L133A 4165 R88L,K118M 5494
E90K,E91A 1508 P27A,Y64A 2837 K105E,L133V 4166 R88L,K118L 5495
E90K,H102A 1509 P27A,K10E 2838 K105E,N95A 4167 R88L,C117S 5496
E90K,H21A 1510 P27A,S17A 2839 K105E,N95V 4168 R88L,C117Q 5497
E90K,K100E 1511 P27A,R122A 2840 K105E,Q127A 4169 R88L,C117T 5498
E90K,K101E 1512 P27A,H124A 2841 K105E,Q127K 4170 R88L,C117P 5499
E90K,K113A 1513 P27A,R122E 2842 K105E,Q43A 4171 R88L,C117A 5500
E90K,K128E 1514 P27A,S47V 2843 K105E,Q43E 4172 R88L,C117I 5501
E90K,L26A 1515 P27A,S17K 2844 K105E,R35A 4173 R88Y,S116W 5502
E90K,L84A 1516 P27A,S116K 2845 K105E,R35E 4174 R88Y,S116Y 5503
E90K,L89A 1517 P27A,N95V 2846 K105E,R35F 4175 R88Y,S116F 5504
E90K,N18R 1518 P27A,Q43K 2847 K105E,R35G 4176 R88Y,S116G 5505
E90K,N92A 1519 P27A,S116H 2848 K105E,R35I 4177 R88Y,S116N 5506
E90K,P121A 1520 P27A,S116Q 2849 K105E,R35L 4178 R88Y,Y15A 5507
E90K,P27A 1521 P27A,K118M 2850 K105E,R35M 4179 R88Y,Y15F 5508
E90K,P79A 1522 P27A,K118L 2851 K105E,R35P 4180 R88Y,Y55A 5509
E90K,Q63A 1523 P27A,C117S 2852 K105E,R35S 4181 R88Y,Y55F 5510
E90K,S38A 1524 P27A,C117Q 2853 K105E,R35V 4182 R88Y,Y94V 5511
E90K,S47A 1525 P27A,C117T 2854 K105E,R37E 4183 R88Y,Y94A 5512
E90K,T30A 1526 P27A,C117P 2855 K105E,R37A 4184 R88Y,E60A 5513
E90K,T34A 1527 P27A,C117A 2856 K105E,R88L 4185 R88Y,Q40A 5514
E90K,T61A 1528 P27A,C117I 2857 K105E,R88Y 4186 R88Y,E60K 5515
E90K,T78A 1529 P79A,Q63A 2858 K105E,R88D 4187 R88Y,Y64A 5516
E90K,T96A 1530 P79A,S38A 2859 K105E,S116W 4188 R88Y,K10E 5517
E90K,C117K 1531 P79A,S47A 2860 K105E,S116Y 4189 R88Y,S17A 5518
E90K,C117R 1532 P79A,T30A 2861 K105E,S116F 4190 R88Y,R122A 5519
E90K,D36K 1533 P79A,T34A 2862 K105E,S116G 4191 R88Y,H124A 5520
E90K,D36A 1534 P79A,T61A 2863 K105E,S116N 4192 R88Y,R122E 5521
E90K,E87K 1535 P79A,T78A 2864 K105E,Y15A 4193 R88Y,S47V 5522
E90K,E87Q 1536 P79A,T96A 2865 K105E,Y15F 4194 R88Y,S17K 5523
E90K,E87V 1537 P79A,C117K 2866 K105E,Y55A 4195 R88Y,S116K 5524
E90K,E87D 1538 P79A,C117R 2867 K105E,Y55F 4196 R88Y,N95V 5525
E90K,K105A 1539 P79A,D36K 2868 K105E,Y94V 4197 R88Y,Q43K 5526
E90K,K105E 1540 P79A,D36A 2869 K105E,Y94A 4198 R88Y,S116H 5527
E90K,K118W 1541 P79A,E87K 2870 K105E,E60A 4199 R88Y,S116Q 5528
E90K,K118H 1542 P79A,E87Q 2871 K105E,Q40A 4200 R88Y,K118M 5529
E90K,K118R 1543 P79A,E87V 2872 K105E,E60K 4201 R88Y,K118L 5530
E90K,K118S 1544 P79A,E87D 2873 K105E,Y64A 4202 R88Y,C117S 5531
E90K,K57A 1545 P79A,K105A 2874 K105E,K10E 4203 R88Y,C117Q 5532
E90K,K57E 1546 P79A,K105E 2875 K105E,S17A 4204 R88Y,C117T 5533
E90K,L133A 1547 P79A,K118W 2876 K105E,R122A 4205 R88Y,C117P 5534
E90K,L133V 1548 P79A,K118H 2877 K105E,H124A 4206 R88Y,C117A 5535
E90K,N95A 1549 P79A,K118R 2878 K105E,R122E 4207 R88Y,C117I 5536
E90K,N95V 1550 P79A,K118S 2879 K105E,S47V 4208 R88D,S116W 5537
E90K,Q127A 1551 P79A,K57A 2880 K105E,S17K 4209 R88D,S116Y 5538 E90K,Q127K 1552 P79A,K57E 2881 K105E,S116K 4210 R88D,S116F 5539
E90K,Q43A 1553 P79A,L133A 2882 K105E,N95V 4211 R88D,S116G 5540
E90K,Q43E 1554 P79A,L133V 2883 K105E,Q43K 4212 R88D,S116N 5541
E90K,R35A 1555 P79A,N95A 2884 K105E,S116H 4213 R88D,Y15A 5542
E90K,R35E 1556 P79A,N95V 2885 K105E,S116Q 4214 R88D,Y15F 5543
E90K,R35F 1557 P79A,Q127A 2886 K105E,K118M 4215 R88D,Y55A 5544
E90K,R35G 1558 P79A,Q127K 2887 K105E,K118L 4216 R88D,Y55F 5545
E90K,R35I 1559 P79A,Q43A 2888 K105E,C117S 4217 R88D,Y94V 5546
E90K,R35L 1560 P79A,Q43E 2889 K105E,C117Q 4218 R88D,Y94A 5547
E90K,R35M 1561 P79A,R35A 2890 K105E,C117T 4219 R88D,E60A 5548
E90K,R35P 1562 P79A,R35E 2891 K105E,C117P 4220 R88D,Q40A 5549
E90K,R35S 1563 P79A,R35F 2892 K105E,C117A 4221 R88D,E60K 5550
E90K,R35V 1564 P79A,R35G 2893 K105E,C117I 4222 R88D,Y64A 5551
E90K,R37E 1565 P79A,R35I 2894 K118W,K57A 4223 R88D,K10E 5552
E90K,R37A 1566 P79A,R35L 2895 K118W,K57E 4224 R88D,S17A 5553
E90K,R88L 1567 P79A,R35M 2896 K118W,L133A 4225 R88D,R122A 5554
E90K,R88Y 1568 P79A,R35P 2897 K118W,L133V 4226 R88D,H124A 5555
E90K,R88D 1569 P79A,R35S 2898 K118W,N95A 4227 R88D,R122E 5556
E90K,S116W 1570 P79A,R35V 2899 K118W,N95V 4228 R88D,S47V 5557
E90K,S116Y 1571 P79A,R37E 2900 K118W,Q127A 4229 R88D,S17K 5558
E90K,S116F 1572 P79A,R37A 2901 K118W,Q127K 4230 R88D,S116K 5559
E90K,S116G 1573 P79A,R88L 2902 K118W,Q43A 4231 R88D,N95V 5560
E90K,S116N 1574 P79A,R88Y 2903 K118W,Q43E 4232 R88D,Q43K 5561
E90K,Y15A 1575 P79A,R88D 2904 K118W,R35A 4233 R88D,S116H 5562
E90K,Y15F 1576 P79A,S116W 2905 K118W,R35E 4234 R88D,S116Q 5563
E90K,Y55A 1577 P79A,S116Y 2906 K118W,R35F 4235 R88D,K118M 5564
E90K,Y55F 1578 P79A,S116F 2907 K118W,R35G 4236 R88D,K118L 5565
E90K,Y94V 1579 P79A,S116G 2908 K118W,R35I 4237 R88D,C117S 5566
E90K,Y94A 1580 P79A,S116N 2909 K118W,R35L 4238 R88D,C117Q 5567
E90K,E60A 1581 P79A,Y15A 2910 K118W,R35M 4239 R88D,C117T 5568
E90K,Q40A 1582 P79A,Y15F 2911 K118W,R35P 4240 R88D,C117P 5569
E90K,E60K 1583 P79A,Y55A 2912 K118W,R35S 4241 R88D,C117A 5570
E90K,Y64A 1584 P79A,Y55F 2913 K118W,R35V 4242 R88D,C117I 5571
E90K,K10E 1585 P79A,Y94V 2914 K118W,R37E 4243 S116W,Y15A 5572
E90K,S17A 1586 P79A,Y94A 2915 K118W,R37A 4244 S116W,Y15F 5573
E90K,R122A 1587 P79A,E60A 2916 K118W,R88L 4245 S116W,Y55A 5574
E90K,H124A 1588 P79A,Q40A 2917 K118W,R88Y 4246 S116W,Y55F 5575
E90K,R122E 1589 P79A,E60K 2918 K118W,R88D 4247 S116W,Y94V 5576
E90K,S47V 1590 P79A,Y64A 2919 K118W,S116W 4248 S116W,Y94A 5577
E90K,S17K 1591 P79A,K10E 2920 K118W,S116Y 4249 S116W,E60A 5578
E90K,S116K 1592 P79A,S17A 2921 K118W,S116F 4250 S116W,Q40A 5579
E90K,N95V 1593 P79A,R122A 2922 K118W,S116G 4251 S116W,E60K 5580
E90K,Q43K 1594 P79A,H124A 2923 K118W,S116N 4252 S116W,Y64A 5581
E90K,S116H 1595 P79A,R122E 2924 K118W,Y15A 4253 S116W,K10E 5582
E90K,S116Q 1596 P79A,S47V 2925 K118W,Y15F 4254 S116W,S17A 5583
E90K,K118M 1597 P79A,S17K 2926 K118W,Y55A 4255 S116W,R122A 5584 E90K,K118L 1598 P79A,S116K 2927 K118W,Y55F 4256 S116W,H124A 5585
E90K,C117S 1599 P79A,N95V 2928 K118W,Y94V 4257 S116W,R122E 5586
E90K,C117Q 1600 P79A,Q43K 2929 K118W,Y94A 4258 S116W,S47V 5587
E90K,C117T 1601 P79A,S116H 2930 K118W,E60A 4259 S116W,S17K 5588
E90K,C117P 1602 P79A,S116Q 2931 K118W,Q40A 4260 S116W,N95V 5589
E90K,C117A 1603 P79A,K118M 2932 K118W,E60K 4261 S116W,Q43K 5590
E90K,C117I 1604 P79A,K118L 2933 K118W,Y64A 4262 S116W,K118M 5591
E91A,H102A 1605 P79A,C117S 2934 K118W,K10E 4263 S116W,K118L 5592
E91A,H21A 1606 P79A,C117Q 2935 K118W,S17A 4264 S116W,C117S 5593
E91A,K100E 1607 P79A,C117T 2936 K118W,R122A 4265 S116W,C117Q 5594
E91A,K101E 1608 P79A,C117P 2937 K118W,H124A 4266 S116W,C117T 5595
E91A,K113A 1609 P79A,C117A 2938 K118W,R122E 4267 S116W,C117P 5596
E91A,K128E 1610 P79A,C117I 2939 K118W,S47V 4268 S116W,C117A 5597
E91A,L26A 1611 Q63A,S38A 2940 K118W,S17K 4269 S116W,C117I 5598
E91A,L84A 1612 Q63A,S47A 2941 K118W,S116K 4270 S116Y,Y15A 5599
E91A,L89A 1613 Q63A,T30A 2942 K118W,N95V 4271 S116Y,Y15F 5600
E91A,N18R 1614 Q63A,T34A 2943 K118W,Q43K 4272 S116Y,Y55A 5601
E91A,N92A 1615 Q63A,T61A 2944 K118W,S116H 4273 S116Y,Y55F 5602
E91A,P121A 1616 Q63A,T78A 2945 K118W,S116Q 4274 S116Y,Y94V 5603
E91A,P27A 1617 Q63A,T96A 2946 K118W,C117S 4275 S116Y,Y94A 5604
E91A,P79A 1618 Q63A,C117K 2947 K118W,C117Q 4276 S116Y,E60A 5605
E91A,Q63A 1619 Q63A,C117R 2948 K118W,C117T 4277 S116Y,Q40A 5606
E91A,S38A 1620 Q63A,D36K 2949 K118W,C117P 4278 S116Y,E60K 5607
E91A,S47A 1621 Q63A,D36A 2950 K118W,C117A 4279 S116Y,Y64A 5608
E91A,T30A 1622 Q63A,E87K 2951 K118W,C117I 4280 S116Y,K10E 5609
E91A,T34A 1623 Q63A,E87Q 2952 K118H,K57A 4281 S116Y,S17A 5610
E91A,T61A 1624 Q63A,E87V 2953 K118H,K57E 4282 S116Y,R122A 5611
E91A,T78A 1625 Q63A,E87D 2954 K118H,L133A 4283 S116Y,H124A 5612
E91A,T96A 1626 Q63A,K105A 2955 K118H,L133V 4284 S116Y,R122E 5613
E91A,C117K 1627 Q63A,K105E 2956 K118H,N95A 4285 S116Y,S47V 5614
E91A,C117R 1628 Q63A,K118W 2957 K118H,N95V 4286 S116Y,S17K 5615
E91A,D36K 1629 Q63A,K118H 2958 K118H,Q127A 4287 S116Y,N95V 5616
E91A,D36A 1630 Q63A,K118R 2959 K118H,Q127K 4288 S116Y,Q43K 5617
E91A,E87K 1631 Q63A,K118S 2960 K118H,Q43A 4289 S116Y,K118M 5618
E91A,E87Q 1632 Q63A,K57A 2961 K118H,Q43E 4290 S116Y,K118L 5619
E91A,E87V 1633 Q63A,K57E 2962 K118H,R35A 4291 S116Y,C117S 5620
E91A,E87D 1634 Q63A,L133A 2963 K118H,R35E 4292 S116Y,C117Q 5621
E91A,K105A 1635 Q63A,L133V 2964 K118H,R35F 4293 S116Y,C117T 5622
E91A,K105E 1636 Q63A,N95A 2965 K118H,R35G 4294 S116Y,C117P 5623
E91A,K118W 1637 Q63A,N95V 2966 K118H,R35I 4295 S116Y,C117A 5624
E91A,K118H 1638 Q63A,Q127A 2967 K118H,R35L 4296 S116Y,C117I 5625
E91A,K118R 1639 Q63A,Q127K 2968 K118H,R35M 4297 S116F,Y15A 5626
E91A,K118S 1640 Q63A,Q43A 2969 K118H,R35P 4298 S116F,Y15F 5627
E91A,K57A 1641 Q63A,Q43E 2970 K118H,R35S 4299 S116F,Y55A 5628
E91A,K57E 1642 Q63A,R35A 2971 K118H,R35V 4300 S116F,Y55F 5629
E91A,L133A 1643 Q63A,R35E 2972 K118H,R37E 4301 S116F,Y94V 5630 E91A,L133V 1644 Q63A,R35F 2973 K118H,R37A 4302 S116F,Y94A 5631
E91A,N95A 1645 Q63A,R35G 2974 K118H,R88L 4303 S116F,E60A 5632
E91A,N95V 1646 Q63A,R35I 2975 K118H,R88Y 4304 S116F,Q40A 5633
E91A,Q127A 1647 Q63A,R35L 2976 K118H,R88D 4305 S116F,E60K 5634
E91A,Q127K 1648 Q63A,R35M 2977 K118H,S116W 4306 S116F,Y64A 5635
E91A,Q43A 1649 Q63A,R35P 2978 K118H,S116Y 4307 S116F,K10E 5636
E91A,Q43E 1650 Q63A,R35S 2979 K118H,S116F 4308 S116F,S17A 5637
E91A,R35A 1651 Q63A,R35V 2980 K118H,S116G 4309 S116F,R122A 5638
E91A,R35E 1652 Q63A,R37E 2981 K118H,S116N 4310 S116F,H124A 5639
E91A,R35F 1653 Q63A,R37A 2982 K118H,Y15A 4311 S116F,R122E 5640
E91A,R35G 1654 Q63A,R88L 2983 K118H,Y15F 4312 S116F,S47V 5641
E91A,R35I 1655 Q63A,R88Y 2984 K118H,Y55A 4313 S116F,S17K 5642
E91A,R35L 1656 Q63A,R88D 2985 K118H,Y55F 4314 S116F,N95V 5643
E91A,R35M 1657 Q63A,S116W 2986 K118H,Y94V 4315 S116F,Q43K 5644
E91A,R35P 1658 Q63A,S116Y 2987 K118H,Y94A 4316 S116F,K118M 5645
E91A,R35S 1659 Q63A,S116F 2988 K118H,E60A 4317 S116F,K118L 5646
E91A,R35V 1660 Q63A,S116G 2989 K118H,Q40A 4318 S116F,C117S 5647
E91A,R37E 1661 Q63A,S116N 2990 K118H,E60K 4319 S116F,C117Q 5648
E91A,R37A 1662 Q63A,Y15A 2991 K118H,Y64A 4320 S116F,C117T 5649
E91A,R88L 1663 Q63A,Y15F 2992 K118H,K10E 4321 S116F,C117P 5650
E91A,R88Y 1664 Q63A,Y55A 2993 K118H,S17A 4322 S116F,C117A 5651
E91A,R88D 1665 Q63A,Y55F 2994 K118H,R122A 4323 S116F,C117I 5652
E91A,S116W 1666 Q63A,Y94V 2995 K118H,H124A 4324 S116G,Y15A 5653
E91A,S116Y 1667 Q63A,Y94A 2996 K118H,R122E 4325 S116G,Y15F 5654
E91A,S116F 1668 Q63A,E60A 2997 K118H,S47V 4326 S116G,Y55A 5655
E91A,S116G 1669 Q63A,Q40A 2998 K118H,S17K 4327 S116G,Y55F 5656
E91A,S116N 1670 Q63A,E60K 2999 K118H,S116K 4328 S116G,Y94V 5657
E91A,Y15A 1671 Q63A,Y64A 3000 K118H,N95V 4329 S116G,Y94A 5658
E91A,Y15F 1672 Q63A,K10E 3001 K118H,Q43K 4330 S116G,E60A 5659
E91A,Y55A 1673 Q63A,S17A 3002 K118H,S116H 4331 S116G,Q40A 5660
E91A,Y55F 1674 Q63A,R122A 3003 K118H,S116Q 4332 S116G,E60K 5661
E91A,Y94V 1675 Q63A,H124A 3004 K118H,C117S 4333 S116G,Y64A 5662
E91A,Y94A 1676 Q63A,R122E 3005 K118H,C117Q 4334 S116G,K10E 5663
E91A,E60A 1677 Q63A,S47V 3006 K118H,C117T 4335 S116G,S17A 5664
E91A,Q40A 1678 Q63A,S17K 3007 K118H,C117P 4336 S116G,R122A 5665
E91A,E60K 1679 Q63A,S116K 3008 K118H,C117A 4337 S116G,H124A 5666
E91A,Y64A 1680 Q63A,N95V 3009 K118H,C117I 4338 S116G,R122E 5667
E91A,K10E 1681 Q63A,Q43K 3010 K118R,K57A 4339 S116G,S47V 5668
E91A,S17A 1682 Q63A,S116H 3011 K118R,K57E 4340 S116G,S17K 5669
E91A,R122A 1683 Q63A,S116Q 3012 K118R,L133A 4341 S116G,N95V 5670
E91A,H124A 1684 Q63A,K118M 3013 K118R,L133V 4342 S116G,Q43K 5671
E91A,R122E 1685 Q63A,K118L 3014 K118R,N95A 4343 S116G,K118M 5672
E91A,S47V 1686 Q63A,C117S 3015 K118R,N95V 4344 S116G,K118L 5673
E91A,S17K 1687 Q63A,C117Q 3016 K118R,Q127A 4345 S116G,C117S 5674
E91A,S116K 1688 Q63A,C117T 3017 K118R,Q127K 4346 S116G,C117Q 5675
E91A,N95V 1689 Q63A,C117P 3018 K118R,Q43A 4347 S116G,C117T 5676 E91A,Q43K 1690 Q63A,C117A 3019 K118R,Q43E 4348 S116G,C117P 5677
E91A,S116H 1691 Q63A,C117I 3020 K118R,R35A 4349 S116G,C117A 5678
E91A,S116Q 1692 S38A,S47A 3021 K118R,R35E 4350 S116G,C117I 5679
E91A,K118M 1693 S38A,T30A 3022 K118R,R35F 4351 S116N,Y15A 5680
E91A,K118L 1694 S38A,T34A 3023 K118R,R35G 4352 S116N,Y15F 5681
E91A,C117S 1695 S38A,T61A 3024 K118R,R35I 4353 S116N,Y55A 5682
E91A,C117Q 1696 S38A,T78A 3025 K118R,R35L 4354 S116N,Y55F 5683
E91A,C117T 1697 S38A,T96A 3026 K118R,R35M 4355 S116N,Y94V 5684
E91A,C117P 1698 S38A,C117K 3027 K118R,R35P 4356 S116N,Y94A 5685
E91A,C117A 1699 S38A,C117R 3028 K118R,R35S 4357 S116N,E60A 5686
E91A,C117I 1700 S38A,D36K 3029 K118R,R35V 4358 S116N,Q40A 5687
H102A,H21A 1701 S38A,D36A 3030 K118R,R37E 4359 S116N,E60K 5688
H102A,K100
1702 S38A,E87K 3031 K118R,R37A 4360 S116N,Y64A 5689 E
H102A,K101
1703 S38A,E87Q 3032 K118R,R88L 4361 S116N,K10E 5690 E
H102A,K113
1704 S38A,E87V 3033 K118R,R88Y 4362 S116N,S17A 5691 A
H102A,K128
1705 S38A,E87D 3034 K118R,R88D 4363 S116N,R122A 5692 E
H102A,L26A 1706 S38A,K105A 3035 K118R,S116W 4364 S116N,H124A 5693
H102A,L84A 1707 S38A,K105E 3036 K118R,S116Y 4365 S116N,R122E 5694
H102A,L89A 1708 S38A,K118W 3037 K118R,S116F 4366 S116N,S47V 5695
H102A,N18R 1709 S38A,K118H 3038 K118R,S116G 4367 S116N,S17K 5696
H102A,N92A 1710 S38A,K118R 3039 K118R,S116N 4368 S116N,N95V 5697
H102A,P121
1711 S38A,K118S 3040 K118R,Y15A 4369 S116N,Q43K 5698 A
H102A,P27A 1712 S38A,K57A 3041 K118R,Y15F 4370 S116N,K118M 5699
H102A,P79A 1713 S38A,K57E 3042 K118R,Y55A 4371 S116N,K118L 5700
H102A,Q63A 1714 S38A,L133A 3043 K118R,Y55F 4372 S116N,C117S 5701
H102A,S38A 1715 S38A,L133V 3044 K118R,Y94V 4373 S116N,C117Q 5702
H102A,S47A 1716 S38A,N95A 3045 K118R,Y94A 4374 S116N,C117T 5703
Η102Α/Γ30Α 1717 S38A,N95V 3046 K118R,E60A 4375 S116N,C117P 5704
H102A,T34A 1718 S38A,Q127A 3047 K118R,Q40A 4376 S116N,C117A 5705
H102A,T61A 1719 S38A,Q127K 3048 K118R,E60K 4377 S116N,C117I 5706
H102A,T78A 1720 S38A,Q43A 3049 K118R,Y64A 4378 Y15A,Y55A 5707
H102A,T96A 1721 S38A,Q43E 3050 K118R,K10E 4379 Y15A,Y55F 5708
H102A,C117
1722 S38A,R35A 3051 K118R,S17A 4380 Y15A,Y94V 5709 K
H102A,C117
1723 S38A,R35E 3052 K118R,R122A 4381 Y15A,Y94A 5710 R
H102A,D36K 1724 S38A,R35F 3053 K118R,H124A 4382 Y15A,E60A 5711
H102A,D36A 1725 S38A,R35G 3054 K118R,R122E 4383 Y15A,Q40A 5712
H102A,E87K 1726 S38A,R35I 3055 K118R,S47V 4384 Y15A,E60K 5713
H102A,E87Q 1727 S38A,R35L 3056 K118R,S17K 4385 Y15A,Y64A 5714
H102A,E87V 1728 S38A,R35M 3057 K118R,S116K 4386 Y15A,K10E 5715
H102A,E87D 1729 S38A,R35P 3058 K118R,N95V 4387 Y15A,S17A 5716
H102A,K105
1730 S38A,R35S 3059 K118R,Q43K 4388 Y15A,R122A 5717 A
H102A,K105 1731 S38A,R35V 3060 K118R,S116H 4389 Y15A,H124A 5718 E
H102A,K118
1732 S38A,R37E 3061 K118R,S116Q 4390 Y15A,R122E 5719 W
H102A,K118
1733 S38A,R37A 3062 K118R,C117S 4391 Y15A,S47V 5720 H
H102A,K118
1734 S38A,R88L 3063 K118R,C117Q 4392 Y15A,S17K 5721 R
H102A,K118
1735 S38A,R88Y 3064 K118R,C117T 4393 Y15A,S116K 5722 S
H102A,K57A 1736 S38A,R88D 3065 K118R,C117P 4394 Y15A,N95V 5723
H102A,K57E 1737 S38A,S116W 3066 K118R,C117A 4395 Y15A,Q43K 5724
H102A,L133
1738 S38A,S116Y 3067 K118R,C117I 4396 Y15A,S116H 5725 A
H102A,L133
1739 S38A,S116F 3068 K118S,K57A 4397 Y15A,S116Q 5726 V
H102A,N95A 1740 S38A,S116G 3069 K118S,K57E 4398 Y15A,K118M 5727
H102A,N95V 1741 S38A,S116N 3070 K118S,L133A 4399 Y15A,K118L 5728
H102A,Q127
1742 S38A,Y15A 3071 K118S,L133V 4400 Y15A,C117S 5729 A
H102A,Q127
1743 S38A,Y15F 3072 K118S,N95A 4401 Y15A,C117Q 5730 K
H102A,Q43A 1744 S38A,Y55A 3073 K118S,N95V 4402 Y15A,C117T 5731
H102A,Q43E 1745 S38A,Y55F 3074 K118S,Q127A 4403 Y15A,C117P 5732
H102A,R35A 1746 S38A,Y94V 3075 K118S,Q127K 4404 Y15A,C117A 5733
H102A,R35E 1747 S38A,Y94A 3076 K118S,Q43A 4405 Y15A,C117I 5734
H102A,R35F 1748 S38A,E60A 3077 K118S,Q43E 4406 Y15F,Y55A 5735
H102A,R35G 1749 S38A,Q40A 3078 K118S,R35A 4407 Y15F,Y55F 5736
H102A,R35I 1750 S38A,E60K 3079 K118S,R35E 4408 Y15F,Y94V 5737
H102A,R35L 1751 S38A,Y64A 3080 K118S,R35F 4409 Y15F,Y94A 5738
H102A,R35M 1752 S38A,K10E 3081 K118S,R35G 4410 Y15F,E60A 5739
H102A,R35P 1753 S38A,S17A 3082 K118S,R35I 4411 Y15F,Q40A 5740
H102A,R35S 1754 S38A,R122A 3083 K118S,R35L 4412 Y15F,E60K 5741
H102A,R35V 1755 S38A,H124A 3084 K118S,R35M 4413 Y15F,Y64A 5742
H102A,R37E 1756 S38A,R122E 3085 K118S,R35P 4414 Y15F,K10E 5743
H102A,R37A 1757 S38A,S47V 3086 K118S,R35S 4415 Y15F,S17A 5744
H102A,R88L 1758 S38A,S17K 3087 K118S,R35V 4416 Y15F,R122A 5745
H102A,R88Y 1759 S38A,S116K 3088 K118S,R37E 4417 Y15F,H124A 5746
H102A,R88D 1760 S38A,N95V 3089 K118S,R37A 4418 Y15F,R122E 5747
H102A,S116
1761 S38A,Q43K 3090 K118S,R88L 4419 Y15F,S47V 5748 W
H102A,S116
1762 S38A,S116H 3091 K118S,R88Y 4420 Y15F,S17K 5749 Y
H102A,S116
1763 S38A,S116Q 3092 K118S,R88D 4421 Y15F,S116K 5750 F
H102A,S116
1764 S38A,K118M 3093 K118S,S116W 4422 Y15F,N95V 5751 G
H102A,S116
1765 S38A,K118L 3094 K118S,S116Y 4423 Y15F,Q43K 5752 N
H102A,Y15A 1766 S38A,C117S 3095 K118S,S116F 4424 Y15F,S116H 5753
H102A,Y15F 1767 S38A,C117Q 3096 K118S,S116G 4425 Y15F,S116Q 5754
H102A,Y55A 1768 S38A,C117T 3097 K118S,S116N 4426 Y15F,K118M 5755
H102A,Y55F 1769 S38A,C117P 3098 K118S,Y15A 4427 Y15F,K118L 5756
Figure imgf000151_0001
H21A,S38A 1809 S47A,R35S 3138 K57A,R35I 4467 Y55F,S17A 5796
H21A,S47A 1810 S47A,R35V 3139 K57A,R35L 4468 Y55F,R122A 5797
H21A,T30A 1811 S47A,R37E 3140 K57A,R35M 4469 Y55F,H124A 5798
H21A,T34A 1812 S47A,R37A 3141 K57A,R35P 4470 Y55F,R122E 5799
H21A,T61A 1813 S47A,R88L 3142 K57A,R35S 4471 Y55F,S47V 5800
H21A,T78A 1814 S47A,R88Y 3143 K57A,R35V 4472 Y55F,S17K 5801
H21A,T96A 1815 S47A,R88D 3144 K57A,R37E 4473 Y55F,S116K 5802
H21A,C117K 1816 S47A,S116W 3145 K57A,R37A 4474 Y55F,N95V 5803
H21A,C117R 1817 S47A,S116Y 3146 K57A,R88L 4475 Y55F,Q43K 5804
H21A,D36K 1818 S47A,S116F 3147 K57A,R88Y 4476 Y55F,S116H 5805
H21A,D36A 1819 S47A,S116G 3148 K57A,R88D 4477 Y55F,S116Q 5806
H21A,E87K 1820 S47A,S116N 3149 K57A,S116W 4478 Y55F,K118M 5807
H21A,E87Q 1821 S47A,Y15A 3150 K57A,S116Y 4479 Y55F,K118L 5808
H21A,E87V 1822 S47A,Y15F 3151 K57A,S116F 4480 Y55F,C117S 5809
H21A,E87D 1823 S47A,Y55A 3152 K57A,S116G 4481 Y55F,C117Q 5810
H21A,K105A 1824 S47A,Y55F 3153 K57A,S116N 4482 Y55F,C117T 5811
H21A,K105E 1825 S47A,Y94V 3154 K57A,Y15A 4483 Y55F,C117P 5812
H21A,K118
1826 S47A,Y94A 3155 K57A,Y15F 4484 Y55F,C117A 5813 W
H21A,K118H 1827 S47A,E60A 3156 K57A,Y55A 4485 Y55F,C117I 5814
H21A,K118R 1828 S47A,Q40A 3157 K57A,Y55F 4486 Y94V,E60A 5815
H21A,K118S 1829 S47A,E60K 3158 K57A,Y94V 4487 Y94V,Q40A 5816
H21A,K57A 1830 S47A,Y64A 3159 K57A,Y94A 4488 Y94V,E60K 5817
H21A,K57E 1831 S47A,K10E 3160 K57A,E60A 4489 Y94V,Y64A 5818
H21A,L133A 1832 S47A,S17A 3161 K57A,Q40A 4490 Y94V,K10E 5819
H21A,L133V 1833 S47A,R122A 3162 K57A,E60K 4491 Y94V,S17A 5820
H21A,N95A 1834 S47A,H124A 3163 K57A,Y64A 4492 Y94V,R122A 5821
H21A,N95V 1835 S47A,R122E 3164 K57A,K10E 4493 Y94V,H124A 5822
H21A,Q127A 1836 S47A,S17K 3165 K57A,S17A 4494 Y94V,R122E 5823
H21A,Q127K 1837 S47A,S116K 3166 K57A,R122A 4495 Y94V,S47V 5824
H21A,Q43A 1838 S47A,N95V 3167 K57A,H124A 4496 Y94V,S17K 5825
H21A,Q43E 1839 S47A,Q43K 3168 K57A,R122E 4497 Y94V,S116K 5826
H21A,R35A 1840 S47A,S116H 3169 K57A,S47V 4498 Y94V,N95V 5827
H21A,R35E 1841 S47A,S116Q 3170 K57A,S17K 4499 Y94V,Q43K 5828
H21A,R35F 1842 S47A,K118M 3171 K57A,S116K 4500 Y94V,S116H 5829
H21A,R35G 1843 S47A,K118L 3172 K57A,N95V 4501 Y94V,S116Q 5830
H21A,R35I 1844 S47A,C117S 3173 K57A,Q43K 4502 Y94V,K118M 5831
H21A,R35L 1845 S47A,C117Q 3174 K57A,S116H 4503 Y94V,K118L 5832
H21A,R35M 1846 S47A,C117T 3175 K57A,S116Q 4504 Y94V,C117S 5833
H21A,R35P 1847 S47A,C117P 3176 K57A,K118M 4505 Y94V,C117Q 5834
H21A,R35S 1848 S47A,C117A 3177 K57A,K118L 4506 Y94V,C117T 5835
H21A,R35V 1849 S47A,C117I 3178 K57A,C117S 4507 Y94V,C117P 5836
H21A,R37E 1850 T30A,T34A 3179 K57A,C117Q 4508 Y94V,C117A 5837
H21A,R37A 1851 T30A,T61A 3180 K57A,C117T 4509 Y94V,C117I 5838
H21A,R88L 1852 T30A,T78A 3181 K57A,C117P 4510 Y94A,E60A 5839
H21A,R88Y 1853 T30A,T96A 3182 K57A,C117A 4511 Y94A,Q40A 5840
H21A,R88D 1854 T30A,C117K 3183 K57A,C117I 4512 Y94A,E60K 5841 H21A,S116W 1855 T30A,C117R 3184 K57E,L133A 4513 Y94A,Y64A 5842
H21A,S116Y 1856 T30A,D36K 3185 K57E,L133V 4514 Y94A,K10E 5843
H21A,S116F 1857 T30A,D36A 3186 K57E,N95A 4515 Y94A,S17A 5844
H21A,S116G 1858 T30A,E87K 3187 K57E,N95V 4516 Y94A,R122A 5845
H21A,S116N 1859 T30A,E87Q 3188 K57E,Q127A 4517 Y94A,H124A 5846
H21A,Y15A 1860 T30A,E87V 3189 K57E,Q127K 4518 Y94A,R122E 5847
H21A,Y15F 1861 T30A,E87D 3190 K57E,Q43A 4519 Y94A,S47V 5848
H21A,Y55A 1862 T30A,K105A 3191 K57E,Q43E 4520 Y94A,S17K 5849
H21A,Y55F 1863 T30A,K105E 3192 K57E,R35A 4521 Y94A,S116K 5850
H21A,Y94V 1864 T30A,K118W 3193 K57E,R35E 4522 Y94A,N95V 5851
H21A,Y94A 1865 T30A,K118H 3194 K57E,R35F 4523 Y94A,Q43K 5852
H21A,E60A 1866 T30A,K118R 3195 K57E,R35G 4524 Y94A,S116H 5853
H21A,Q40A 1867 T30A,K118S 3196 K57E,R35I 4525 Y94A,S116Q 5854
H21A,E60K 1868 T30A,K57A 3197 K57E,R35L 4526 Y94A,K118M 5855
H21A,Y64A 1869 T30A,K57E 3198 K57E,R35M 4527 Y94A,K118L 5856
H21A,K10E 1870 T30A,L133A 3199 K57E,R35P 4528 Y94A,C117S 5857
H21A,S17A 1871 T30A,L133V 3200 K57E,R35S 4529 Y94A,C117Q 5858
H21A,R122A 1872 T30A,N95A 3201 K57E,R35V 4530 Y94A,C117T 5859
H21A,H124A 1873 T30A,N95V 3202 K57E,R37E 4531 Y94A,C117P 5860
H21A,R122E 1874 T30A,Q127A 3203 K57E,R37A 4532 Y94A,C117A 5861
H21A,S47V 1875 T30A,Q127K 3204 K57E,R88L 4533 Y94A,C117I 5862
H21A,S17K 1876 T30A,Q43A 3205 K57E,R88Y 4534 E60A,Q40A 5863
H21A,S116K 1877 T30A,Q43E 3206 K57E,R88D 4535 E60A,Y64A 5864
H21A,N95V 1878 T30A,R35A 3207 K57E,S116W 4536 E60A,K10E 5865
H21A,Q43K 1879 T30A,R35E 3208 K57E,S116Y 4537 E60A,S17A 5866
H21A,S116H 1880 T30A,R35F 3209 K57E,S116F 4538 E60A,R122A 5867
H21A,S116Q 1881 T30A,R35G 3210 K57E,S116G 4539 E60A,H124A 5868
H21A,K118
1882 T30A,R35I 3211 K57E,S116N 4540 E60A,R122E 5869 M
H21A,K118L 1883 T30A,R35L 3212 K57E,Y15A 4541 E60A,S47V 5870
H21A,C117S 1884 T30A,R35M 3213 K57E,Y15F 4542 E60A,S17K 5871
H21A,C117Q 1885 T30A,R35P 3214 K57E,Y55A 4543 E60A,S116K 5872
H21A,C117T 1886 T30A,R35S 3215 K57E,Y55F 4544 E60A,N95V 5873
H21A,C117P 1887 T30A,R35V 3216 K57E,Y94V 4545 E60A,Q43K 5874
H21A,C117A 1888 T30A,R37E 3217 K57E,Y94A 4546 E60A,S116H 5875
H21A,C117I 1889 T30A,R37A 3218 K57E,E60A 4547 E60A,S116Q 5876
K100E,K101
1890 T30A,R88L 3219 K57E,Q40A 4548 E60A,K118M 5877 E
K100E,K113
1891 T30A,R88Y 3220 K57E,E60K 4549 E60A,K118L 5878 A
K100E,K128
1892 T30A,R88D 3221 K57E,Y64A 4550 E60A,C117S 5879 E
K100E,L26A 1893 T30A,S116W 3222 K57E,K10E 4551 E60A,C117Q 5880
K100E,L84A 1894 T30A,S116Y 3223 K57E,S17A 4552 E60A,C117T 5881
K100E,L89A 1895 T30A,S116F 3224 K57E,R122A 4553 E60A,C117P 5882
K100E,N18R 1896 T30A,S116G 3225 K57E,H124A 4554 E60A,C117A 5883
K100E,N92A 1897 T30A,S116N 3226 K57E,R122E 4555 E60A,C117I 5884
K100E,P121 1898 T30A,Y15A 3227 K57E,S47V 4556 Q40A,E60K 5885 A
K100E,P27A 1899 T30A,Y15F 3228 K57E,S17K 4557 Q40A,Y64A 5886
K100E,P79A 1900 T30A,Y55A 3229 K57E,S116K 4558 Q40A,K10E 5887
K100E,Q63A 1901 T30A,Y55F 3230 K57E,N95V 4559 Q40A,S17A 5888
K100E,S38A 1902 T30A,Y94V 3231 K57E,Q43K 4560 Q40A,R122A 5889
K100E,S47A 1903 T30A,Y94A 3232 K57E,S116H 4561 Q40A,H124A 5890
K100E,T30A 1904 T30A,E60A 3233 K57E,S116Q 4562 Q40A,R122E 5891
K100E,T34A 1905 T30A,Q40A 3234 K57E,K118M 4563 Q40A,S47V 5892
K100E,T61A 1906 T30A,E60K 3235 K57E,K118L 4564 Q40A,S17K 5893
K100E,T78A 1907 T30A,Y64A 3236 K57E,C117S 4565 Q40A,S116K 5894
K100E,T96A 1908 T30A,K10E 3237 K57E,C117Q 4566 Q40A,N95V 5895
K100E,C117
1909 T30A,S17A 3238 K57E,C117T 4567 Q40A,Q43K 5896 K
K100E,C117
1910 T30A,R122A 3239 K57E,C117P 4568 Q40A,S116H 5897 R
K100E,D36K 1911 T30A,H124A 3240 K57E,C117A 4569 Q40A,S116Q 5898
K100E,D36A 1912 T30A,R122E 3241 K57E,C117I 4570 Q40A,K118M 5899
K100E,E87K 1913 T30A,S47V 3242 L133A,N95A 4571 Q40A,K118L 5900
K100E,E87Q 1914 T30A,S17K 3243 L133A,N95V 4572 Q40A,C117S 5901
K100E,E87V 1915 T30A,S116K 3244 L133A,Q127A 4573 Q40A,C117Q 5902
K100E,E87D 1916 T30A,N95V 3245 L133A,Q127K 4574 Q40A,C117T 5903
K100E,K105
1917 T30A,Q43K 3246 L133A,Q43A 4575 Q40A,C117P 5904 A
K100E,K105
1918 T30A,S116H 3247 L133A,Q43E 4576 Q40A,C117A 5905 E
K100E,K118
1919 T30A,S116Q 3248 L133A,R35A 4577 Q40A,C117I 5906 W
K100E,K118
1920 T30A,K118M 3249 L133A,R35E 4578 E60K,Y64A 5907 H
K100E,K118
1921 T30A,K118L 3250 L133A,R35F 4579 E60K,K10E 5908 R
K100E,K118
1922 T30A,C117S 3251 L133A,R35G 4580 E60K,S17A 5909 s
K100E,K57A 1923 T30A,C117Q 3252 L133A,R35I 4581 E60K,R122A 5910
K100E,K57E 1924 T30A,C117T 3253 L133A,R35L 4582 E60K,H124A 5911
K100E,L133
1925 T30A,C117P 3254 L133A,R35M 4583 E60K,R122E 5912 A
K100E,L133
1926 T30A,C117A 3255 L133A,R35P 4584 E60K,S47V 5913 V
K100E,N95A 1927 T30A,C117I 3256 L133A,R35S 4585 E60K,S17K 5914
K100E,N95V 1928 T34A,T61A 3257 L133A,R35V 4586 E60K,S116K 5915
K100E,Q127
1929 T34A,T78A 3258 L133A,R37E 4587 E60K,N95V 5916 A
K100E,Q127
1930 T34A,T96A 3259 L133A,R37A 4588 E60K,Q43K 5917 K
K100E,Q43A 1931 T34A,C117K 3260 L133A,R88L 4589 E60K,S116H 5918
K100E,Q43E 1932 T34A,C117R 3261 L133A,R88Y 4590 E60K,S116Q 5919
K100E,R35A 1933 T34A,D36K 3262 L133A,R88D 4591 E60K,K118M 5920
K100E,R35E 1934 T34A,D36A 3263 L133A,S116W 4592 E60K,K118L 5921
K100E,R35F 1935 T34A,E87K 3264 L133A,S116Y 4593 E60K,C117S 5922
K100E,R35G 1936 T34A,E87Q 3265 L133A,S116F 4594 E60K,C117Q 5923
Figure imgf000155_0001
Figure imgf000156_0001
K101E,K118
2014 T61A,E87D 3343 L133V,Q43K 4672 H124A,S47V 6001 s
K101E,K57A 2015 T61A,K105A 3344 L133V,S116H 4673 H124A,S17K 6002
K101E,K57E 2016 T61A,K105E 3345 L133V,S116Q 4674 H124A,S116K 6003
K101E,L133
2017 T61A,K118W 3346 L133V,K118M 4675 H124A,N95V 6004 A
K101E,L133
2018 T61A,K118H 3347 L133V,K118L 4676 H124A,Q43K 6005 V
K101E,N95A 2019 T61A,K118R 3348 L133V,C117S 4677 H124A,S116H 6006
K101E,N95V 2020 T61A,K118S 3349 L133V,C117Q 4678 H124A,S116Q 6007
K101E,Q127
2021 T61A,K57A 3350 L133V,C117T 4679 H124A,K118M 6008 A
K101E,Q127
2022 T61A,K57E 3351 L133V,C117P 4680 H124A,K118L 6009 K
K101E,Q43A 2023 T61A,L133A 3352 L133V,C117A 4681 H124A,C117S 6010
K101E,Q43E 2024 T61A,L133V 3353 L133V,C117I 4682 H124A,C117Q 6011
K101E,R35A 2025 T61A,N95A 3354 N95A,Q127A 4683 H124A,C117T 6012
K101E,R35E 2026 T61A,N95V 3355 N95A,Q127K 4684 H124A,C117P 6013
K101E,R35F 2027 T61A,Q127A 3356 N95A,Q43A 4685 H124A,C117A 6014
K101E,R35G 2028 T61A,Q127K 3357 N95A,Q43E 4686 H124A,C117I 6015
K101E,R35I 2029 T61A,Q43A 3358 N95A,R35A 4687 R122E,S47V 6016
K101E,R35L 2030 T61A,Q43E 3359 N95A,R35E 4688 R122E,S17K 6017
K101E,R35M 2031 T61A,R35A 3360 N95A,R35F 4689 R122E,S116K 6018
K101E,R35P 2032 T61A,R35E 3361 N95A,R35G 4690 R122E,N95V 6019
K101E,R35S 2033 T61A,R35F 3362 N95A,R35I 4691 R122E,Q43K 6020
K101E,R35V 2034 T61A,R35G 3363 N95A,R35L 4692 R122E,S116H 6021
K101E,R37E 2035 T61A,R35I 3364 N95A,R35M 4693 R122E,S116Q 6022
K101E,R37A 2036 T61A,R35L 3365 N95A,R35P 4694 R122E,K118M 6023
K101E,R88L 2037 T61A,R35M 3366 N95A,R35S 4695 R122E,K118L 6024
K101E,R88Y 2038 T61A,R35P 3367 N95A,R35V 4696 R122E,C117S 6025
K101E,R88D 2039 T61A,R35S 3368 N95A,R37E 4697 R122E,C117Q 6026
K101E,S116
2040 T61A,R35V 3369 N95A,R37A 4698 R122E,C117T 6027 W
K101E,S116
2041 T61A,R37E 3370 N95A,R88L 4699 R122E,C117P 6028 Y
K101E,S116F 2042 T61A,R37A 3371 N95A,R88Y 4700 R122E,C117A 6029
K101E,S116
2043 T61A,R88L 3372 N95A,R88D 4701 R122E,C117I 6030 G
K101E,S116
2044 T61A,R88Y 3373 N95A,S116W 4702 S47V,S17K 6031 N
K101E,Y15A 2045 T61A,R88D 3374 N95A,S116Y 4703 S47V,S116K 6032
K101E,Y15F 2046 T61A,S116W 3375 N95A,S116F 4704 S47V,N95V 6033
K101E,Y55A 2047 T61A,S116Y 3376 N95A,S116G 4705 S47V,Q43K 6034
K101E,Y55F 2048 T61A,S116F 3377 N95A,S116N 4706 S47V,S116H 6035
K101E,Y94V 2049 T61A,S116G 3378 N95A,Y15A 4707 S47V,S116Q 6036
K101E,Y94A 2050 T61A,S116N 3379 N95A,Y15F 4708 S47V,K118M 6037
K101E,E60A 2051 T61A,Y15A 3380 N95A,Y55A 4709 S47V,K118L 6038
K101E,Q40A 2052 T61A,Y15F 3381 N95A,Y55F 4710 S47V,C117S 6039
K101E,E60K 2053 T61A,Y55A 3382 N95A,Y94V 4711 S47V,C117Q 6040
K101E,Y64A 2054 T61A,Y55F 3383 N95A,Y94A 4712 S47V,C117T 6041
Figure imgf000158_0001
κ
K113A,C117
2093 T78A,K118H 3422 N95V,R37A 4751 Q43K,S116Q 6080 R
K113A,D36K 2094 T78A,K118R 3423 N95V,R88L 4752 Q43K,K118M 6081
K113A,D36A 2095 T78A,K118S 3424 N95V,R88Y 4753 Q43K,K118L 6082
K113A,E87K 2096 T78A,K57A 3425 N95V,R88D 4754 Q43K,C117S 6083
K113A,E87Q 2097 T78A,K57E 3426 N95V,S116W 4755 Q43K,C117Q 6084
K113A,E87V 2098 T78A,L133A 3427 N95V,S116Y 4756 Q43K,C117T 6085
K113A,E87D 2099 T78A,L133V 3428 N95V,S116F 4757 Q43K,C117P 6086
K113A,K105
2100 T78A,N95A 3429 N95V,S116G 4758 Q43K,C117A 6087 A
K113A,K105
2101 T78A,N95V 3430 N95V,S116N 4759 Q43K,C117I 6088 E
K113A,K118
2102 T78A,Q127A 3431 N95V,Y15A 4760 S116H,K118M 6089 W
K113A,K118
2103 T78A,Q127K 3432 N95V,Y15F 4761 S116H,K118L 6090 H
K113A,K118
2104 T78A,Q43A 3433 N95V,Y55A 4762 S116H,C117S 6091 R
K113A,K118
2105 T78A,Q43E 3434 N95V,Y55F 4763 S116H,C117Q 6092 S
K113A,K57A 2106 T78A,R35A 3435 N95V,Y94V 4764 S116H,C117T 6093
K113A,K57E 2107 T78A,R35E 3436 N95V,Y94A 4765 S116H,C117P 6094
K113A,L133
2108 T78A,R35F 3437 N95V,E60A 4766 S116H,C117A 6095 A
K113A,L133
2109 T78A,R35G 3438 N95V,Q40A 4767 S116H,C117I 6096 V
K113A,N95A 2110 T78A,R35I 3439 N95V,E60K 4768 S116Q,K118M 6097
K113A,N95V 2111 T78A,R35L 3440 N95V,Y64A 4769 S116Q,K118L 6098
K113A,Q127
2112 T78A,R35M 3441 N95V,K10E 4770
A S116Q,C117S 6099
K113A,Q127
2113 T78A,R35P 3442 N95V,S17A 4771 S116Q,C117Q 6100 K
K113A,Q43A 2114 T78A,R35S 3443 N95V,R122A 4772 S116Q,C117T 6101
K113A,Q43E 2115 T78A,R35V 3444 N95V,H124A 4773 S116Q,C117P 6102
K113A,R35A 2116 T78A,R37E 3445 N95V,R122E 4774 S116Q,C117A 6103
K113A,R35E 2117 T78A,R37A 3446 N95V,S47V 4775 S116Q,C117I 6104
K113A,R35F 2118 T78A,R88L 3447 N95V,S17K 4776 K118M,C117S 6105
K113A,R35G 2119 T78A,R88Y 3448 N95V,S116K 4777 K118M,C117Q 6106
K113A,R35I 2120 T78A,R88D 3449 N95V,Q43K 4778 K118M,C117T 6107
K113A,R35L 2121 T78A,S116W 3450 N95V,S116H 4779 K118M,C117P 6108
K113A,R35M 2122 T78A,S116Y 3451 N95V,S116Q 4780 K118M,C117A 6109
K113A,R35P 2123 T78A,S116F 3452 N95V,K118M 4781 K118M,C117I 6110
K113A,R35S 2124 T78A,S116G 3453 N95V,K118L 4782 K118L,C117S 6111
K113A,R35V 2125 T78A,S116N 3454 N95V,C117S 4783 K118L,C117Q 6112
K113A,R37E 2126 T78A,Y15A 3455 N95V,C117Q 4784 K118L,C117T 6113
K113A,R37A 2127 T78A,Y15F 3456 N95V,C117T 4785 K118L,C117P 6114
K113A,R88L 2128 T78A,Y55A 3457 N95V,C117P 4786 K118L,C117A 6115
K113A,R88Y 2129 T78A,Y55F 3458 N95V,C117A 4787 K118L,C117I 6116
K113A,R88D 2130 T78A,Y94V 3459 N95V,C117I 4788
K113A,S116 2131 T78A,Y94A 3460 Q127A,Q43A 4789 w
Table 11.
Figure imgf000160_0001
R37D,S38A 6158 Y15S,Y94V 11028 N95S,C117Q 15898 L84G,K57A 20768
R37D,D39A 6159 Y15S,Y94A 11029 N95S,C117T 15899 L84G,K57E 20769
R37D,Q40A 6160 Y15S,N95A 11030 N95S,C117P 15900 L84G,E60A 20770
R37D,Q43A 6161 Y15S,N95V 11031 N95S,C117A 15901 L84G,E60K 20771
R37D,Q43E 6162 Y15S,T96A 11032 N95S,C117I 15902 L84G,T61A 20772
N95S,K118
R37D,Q43K 6163 Y15S,K100E 11033 15903 L84G,Q63A 20773
W
R37D,S47A 6164 Y15S,K101E 11034 N95S,K118H 15904 L84G,Y64A 20774
R37D,S47V 6165 Y15S,H102A 11035 N95S,K118R 15905 L84G,D68A 20775
R37D,A48G 6166 Y15S,K105A 11036 N95S,K118S 15906 L84G,T78A 20776
N95S,K118
R37D,E53A 6167 Y15S,K105E 11037 15907 L84G,P79A 20777
M
R37D,Y55A 6168 Y15S,K113A 11038 N95S,K118L 15908 L84G,E81K 20778
R37D,Y55F 6169 Y15S,S116W 11039 N95S,P121A 15909 L84G,E87K 20779
R37D,K57A 6170 Y15S,S116Y 11040 N95S,R122A 15910 L84G,E87Q 20780
R37D,K57E 6171 Y15S,S116F 11041 N95S,R122E 15911 L84G,E87V 20781
R37D,E60A 6172 Y15S,S116G 11042 N95S,H124A 15912 L84G,E87D 20782
R37D,E60K 6173 Y15S,S116N 11043 N95S,Q127A 15913 L84G,R88L 20783
R37D,T61A 6174 Y15S,S116K 11044 N95S,Q127K 15914 L84G,R88Y 20784
R37D,Q63A 6175 Y15S,S116H 11045 N95S,K128E 15915 L84G,R88D 20785
R37D,Y64A 6176 Y15S,S116Q 11046 N95S,L133A 15916 L84G,L89A 20786
R37D,D68A 6177 Y15S,C117K 11047 N95S,L133V 15917 L84G,E90K 20787
R37D,T78A 6178 Y15S,C117R 11048 N95T,K10E 15918 L84G,E91A 20788
R37D,P79A 6179 Y15S,C117S 11049 N95T,Y15A 15919 L84G,N92A 20789
R37D,E81K 6180 Y15S,C117Q 11050 N95T,Y15F 15920 L84G,Y94V 20790
R37D,L84A 6181 Y15S,C117T 11051 N95T,S17A 15921 L84G,Y94A 20791
R37D,E87K 6182 Y15S,C117P 11052 N95T,S17K 15922 L84G,N95A 20792
R37D,E87Q 6183 Y15S,C117A 11053 N95T,N18R 15923 L84G,N95V 20793
R37D,E87V 6184 Y15S,C117I 11054 N95T,H21A 15924 L84G,T96A 20794
R37D,E87D 6185 Y15S,K118W 11055 N95T,L26A 15925 L84G,K100E 20795
R37D,R88L 6186 Y15S,K118H 11056 N95T,P27A 15926 L84G,K101E 20796
R37D,R88Y 6187 Y15S,K118R 11057 N95T,D28K 15927 L84G,H102A 20797
R37D,R88D 6188 Y15S,K118S 11058 N95T,T30A 15928 L84G,K105A 20798
R37D,L89A 6189 Y15S,K118M 11059 N95T,T34A 15929 L84G,K105E 20799
R37D,E90K 6190 Y15S,K118L 11060 N95T,R35A 15930 L84G,K113A 20800
R37D,E91A 6191 Y15S,P121A 11061 N95T,R35E 15931 L84G,S116W 20801
R37D,N92A 6192 Y15S,R122A 11062 N95T,R35F 15932 L84G,S116Y 20802
R37D,Y94V 6193 Y15S,R122E 11063 N95T,R35G 15933 L84G,S116F 20803
R37D,Y94A 6194 Y15S,H124A 11064 N95T,R35I 15934 L84G,S116G 20804
R37D,N95A 6195 Y15S,Q127A 11065 N95T,R35L 15935 L84G,S116N 20805
R37D,N95V 6196 Y15S,Q127K 11066 N95T,R35M 15936 L84G,S116K 20806
R37D,T96A 6197 Y15S,K128E 11067 N95T,R35P 15937 L84G,S116H 20807
R37D,K100E 6198 Y15S,L133A 11068 N95T,R35S 15938 L84G,S116Q 20808
R37D,K101E 6199 Y15S,L133V 11069 N95T,R35V 15939 L84G,C117K 20809
R37D,H102A 6200 Y15T,K10E 11070 N95T,D36A 15940 L84G,C117R 20810
R37D,K105A 6201 Y15T,S17A 11071 N95T,D36K 15941 L84G,C117S 20811
R37D,K105E 6202 Y15T,S17K 11072 N95T,R37E 15942 L84G,C117Q 20812 R37D,K113A 6203 Y15T,N18R 11073 N95T,R37A 15943 L84G,C117T 20813
R37D,S116W 6204 Y15T,H21A 11074 N95T,S38A 15944 L84G,C117P 20814
R37D,S116Y 6205 Y15T,L26A 11075 N95T,D39A 15945 L84G,C117A 20815
R37D,S116F 6206 Y15T,P27A 11076 N95T,Q40A 15946 L84G,C117I 20816
R37D,S116G 6207 Y15T,D28K 11077 N95T,Q43A 15947 L84G,K118W 20817
R37D,S116N 6208 Y15T,T30A 11078 N95T,Q43E 15948 L84G,K118H 20818
R37D,S116K 6209 Y15T,T34A 11079 N95T,Q43K 15949 L84G,K118R 20819
R37D,S116H 6210 Y15T,R35A 11080 N95T,S47A 15950 L84G,K118S 20820
R37D,S116Q 6211 Y15T,R35E 11081 N95T,S47V 15951 L84G,K118M 20821
R37D,C117K 6212 Y15T,R35F 11082 N95T,A48G 15952 L84G,K118L 20822
R37D,C117R 6213 Y15T,R35G 11083 N95T,E53A 15953 L84G,P121A 20823
R37D,C117S 6214 Y15T,R35I 11084 N95T,Y55A 15954 L84G,R122A 20824
R37D,C117Q 6215 Y15T,R35L 11085 N95T,Y55F 15955 L84G,R122E 20825
R37D,C117T 6216 Y15T,R35M 11086 N95T,K57A 15956 L84G,H124A 20826
R37D,C117P 6217 Y15T,R35P 11087 N95T,K57E 15957 L84G,Q127A 20827
R37D,C117A 6218 Y15T,R35S 11088 N95T,E60A 15958 L84G,Q127K 20828
R37D,C117I 6219 Y15T,R35V 11089 N95T,E60K 15959 L84G,K128E 20829
R37D,K118
6220 Y15T,D36A 11090 N95T,T61A 15960 L84G,L133A 20830 W
R37D,K118H 6221 Y15T,D36K 11091 N95T,Q63A 15961 L84G,L133V 20831
R37D,K118R 6222 Y15T,R37E 11092 N95T,Y64A 15962 L84H,K10E 20832
R37D,K118S 6223 Y15T,R37A 11093 N95T,D68A 15963 L84H,Y15A 20833
R37D,K118
6224 Y15T,S38A 11094 N95T,T78A 15964 L84H,Y15F 20834 M
R37D,K118L 6225 Y15T,D39A 11095 N95T,P79A 15965 L84H,S17A 20835
R37D,P121A 6226 Y15T,Q40A 11096 N95T,E81K 15966 L84H,S17K 20836
R37D,R122A 6227 Y15T,Q43A 11097 N95T,L84A 15967 L84H,N18R 20837
R37D,R122E 6228 Y15T,Q43E 11098 N95T,E87K 15968 L84H,H21A 20838
R37D,H124A 6229 Y15T,Q43K 11099 N95T,E87Q 15969 L84H,L26A 20839
R37D,Q127A 6230 Y15T,S47A 11100 N95T,E87V 15970 L84H,P27A 20840
R37D,Q127K 6231 Y15T,S47V 11101 N95T,E87D 15971 L84H,D28K 20841
R37D,K128E 6232 Y15T,A48G 11102 N95T,R88L 15972 L84H,T30A 20842
R37D,L133A 6233 Y15T,E53A 11103 N95T,R88Y 15973 L84H,T34A 20843
R37D,L133V 6234 Y15T,Y55A 11104 N95T,R88D 15974 L84H,R35A 20844
R37F,Q43D 6235 Y15T,Y55F 11105 N95T,L89A 15975 L84H,R35E 20845
R37F,Q43F 6236 Y15T,K57A 11106 N95T,E90K 15976 L84H,R35F 20846
R37F,Q43G 6237 Y15T,K57E 11107 N95T,E91A 15977 L84H,R35G 20847
R37F,Q43H 6238 Y15T,E60A 11108 N95T,N92A 15978 L84H,R35I 20848
R37F,Q43I 6239 Y15T,E60K 11109 N95T,Y94V 15979 L84H,R35L 20849
R37F,Q43L 6240 Y15T,T61A 11110 N95T,Y94A 15980 L84H,R35M 20850
R37F,Q43M 6241 Y15T,Q63A 11111 N95T,T96A 15981 L84H,R35P 20851
R37F,Q43N 6242 Y15T,Y64A 11112 N95T,K100E 15982 L84H,R35S 20852
R37F,Q43P 6243 Y15T,D68A 11113 N95T,K101E 15983 L84H,R35V 20853
R37F,Q43S 6244 Y15T,T78A 11114 N95T,H102A 15984 L84H,D36A 20854
R37F,Q43T 6245 Y15T,P79A 11115 N95T,K105A 15985 L84H,D36K 20855
R37F,Q43V 6246 Y15T,E81K 11116 N95T,K105E 15986 L84H,R37E 20856
R37F,Q43W 6247 Y15T,L84A 11117 N95T,K113A 15987 L84H,R37A 20857 R37F,Q43Y 6248 Y15T,E87K 11118 N95T,S116W 15988 L84H,S38A 20858
R37F,D28A 6249 Y15T,E87Q 11119 N95T,S116Y 15989 L84H,D39A 20859
R37F,E49A 6250 Y15T,E87V 11120 N95T,S116F 15990 L84H,Q40A 20860
R37F,P121A 6251 Y15T,E87D 11121 N95T,S116G 15991 L84H,Q43A 20861
R37F,K10E 6252 Y15T,R88L 11122 N95T,S116N 15992 L84H,Q43E 20862
R37F,Y15A 6253 Y15T,R88Y 11123 N95T,S116K 15993 L84H,Q43K 20863
R37F,Y15F 6254 Y15T,R88D 11124 N95T,S116H 15994 L84H,S47A 20864
R37F,S17A 6255 Y15T,L89A 11125 N95T,S116Q 15995 L84H,S47V 20865
R37F,S17K 6256 Y15T,E90K 11126 N95T,C117K 15996 L84H,A48G 20866
R37F,N18R 6257 Y15T,E91A 11127 N95T,C117R 15997 L84H,E53A 20867
R37F,H21A 6258 Y15T,N92A 11128 N95T,C117S 15998 L84H,Y55A 20868
R37F,L26A 6259 Y15T,Y94V 11129 N95T,C117Q 15999 L84H,Y55F 20869
R37F,P27A 6260 Y15T,Y94A 11130 N95T,C117T 16000 L84H,K57A 20870
R37F,D28K 6261 Y15T,N95A 11131 N95T,C117P 16001 L84H,K57E 20871
R37F,T30A 6262 Y15T,N95V 11132 N95T,C117A 16002 L84H,E60A 20872
R37F,T34A 6263 Y15T,T96A 11133 N95T,C117I 16003 L84H,E60K 20873
N95T,K118
R37F,R35A 6264 Y15T,K100E 11134 16004 L84H,T61A 20874
W
R37F,R35E 6265 Y15T,K101E 11135 N95T,K118H 16005 L84H,Q63A 20875
R37F,R35F 6266 Y15T,H102A 11136 N95T,K118R 16006 L84H,Y64A 20876
R37F,R35G 6267 Y15T,K105A 11137 N95T,K118S 16007 L84H,D68A 20877
N95T,K118
R37F,R35I 6268 Y15T,K105E 11138 16008 L84H,T78A 20878
M
R37F,R35L 6269 Y15T,K113A 11139 N95T,K118L 16009 L84H,P79A 20879
R37F,R35M 6270 Y15T,S116W 11140 N95T,P121A 16010 L84H,E81K 20880
R37F,R35P 6271 Y15T,S116Y 11141 N95T,R122A 16011 L84H,E87K 20881
R37F,R35S 6272 Y15T,S116F 11142 N95T,R122E 16012 L84H,E87Q 20882
R37F,R35V 6273 Y15T,S116G 11143 N95T,H124A 16013 L84H,E87V 20883
R37F,D36A 6274 Y15T,S116N 11144 N95T,Q127A 16014 L84H,E87D 20884
R37F,D36K 6275 Y15T,S116K 11145 N95T,Q127K 16015 L84H,R88L 20885
R37F,S38A 6276 Y15T,S116H 11146 N95T,K128E 16016 L84H,R88Y 20886
R37F,D39A 6277 Y15T,S116Q 11147 N95T,L133A 16017 L84H,R88D 20887
R37F,Q40A 6278 Y15T,C117K 11148 N95T,L133V 16018 L84H,L89A 20888
R37F,Q43A 6279 Y15T,C117R 11149 N95W,K10E 16019 L84H,E90K 20889
R37F,Q43E 6280 Y15T,C117S 11150 N95W,Y15A 16020 L84H,E91A 20890
R37F,Q43K 6281 Y15T,C117Q 11151 N95W,Y15F 16021 L84H,N92A 20891
R37F,S47A 6282 Y15T,C117T 11152 N95W,S17A 16022 L84H,Y94V 20892
R37F,S47V 6283 Y15T,C117P 11153 N95W,S17K 16023 L84H,Y94A 20893
R37F,A48G 6284 Y15T,C117A 11154 N95W,N18R 16024 L84H,N95A 20894
R37F,E53A 6285 Y15T,C117I 11155 N95W,H21A 16025 L84H,N95V 20895
R37F,Y55A 6286 Y15T,K118W 11156 N95W,L26A 16026 L84H,T96A 20896
R37F,Y55F 6287 Y15T,K118H 11157 N95W,P27A 16027 L84H,K100E 20897
R37F,K57A 6288 Y15T,K118R 11158 N95W,D28K 16028 L84H,K101E 20898
R37F,K57E 6289 Y15T,K118S 11159 N95W,T30A 16029 L84H,H102A 20899
R37F,E60A 6290 Y15T,K118M 11160 N95W,T34A 16030 L84H,K105A 20900
R37F,E60K 6291 Y15T,K118L 11161 N95W,R35A 16031 L84H,K105E 20901
R37F,T61A 6292 Y15T,P121A 11162 N95W,R35E 16032 L84H,K113A 20902 R37F,Q63A 6293 Y15T,R122A 11163 N95W,R35F 16033 L84H,S116W 20903
R37F,Y64A 6294 Y15T,R122E 11164 N95W,R35G 16034 L84H,S116Y 20904
R37F,D68A 6295 Y15T,H124A 11165 N95W,R35I 16035 L84H,S116F 20905
R37F,T78A 6296 Y15T,Q127A 11166 N95W,R35L 16036 L84H,S116G 20906
R37F,P79A 6297 Y15T,Q127K 11167 N95W,R35M 16037 L84H,S116N 20907
R37F,E81K 6298 Y15T,K128E 11168 N95W,R35P 16038 L84H,S116K 20908
R37F,L84A 6299 Y15T,L133A 11169 N95W,R35S 16039 L84H,S116H 20909
R37F,E87K 6300 Y15T,L133V 11170 N95W,R35V 16040 L84H,S116Q 20910
R37F,E87Q 6301 Y15V,K10E 11171 N95W,D36A 16041 L84H,C117K 20911
R37F,E87V 6302 Y15V,S17A 11172 N95W,D36K 16042 L84H,C117R 20912
R37F,E87D 6303 Y15V,S17K 11173 N95W,R37E 16043 L84H,C117S 20913
R37F,R88L 6304 Y15V,N18R 11174 N95W,R37A 16044 L84H,C117Q 20914
R37F,R88Y 6305 Y15V,H21A 11175 N95W,S38A 16045 L84H,C117T 20915
R37F,R88D 6306 Y15V,L26A 11176 N95W,D39A 16046 L84H,C117P 20916
R37F,L89A 6307 Y15V,P27A 11177 N95W,Q40A 16047 L84H,C117A 20917
R37F,E90K 6308 Y15V,D28K 11178 N95W,Q43A 16048 L84H,C117I 20918
R37F,E91A 6309 Y15V,T30A 11179 N95W,Q43E 16049 L84H,K118W 20919
R37F,N92A 6310 Y15V,T34A 11180 N95W,Q43K 16050 L84H,K118H 20920
R37F,Y94V 6311 Y15V,R35A 11181 N95W,S47A 16051 L84H,K118R 20921
R37F,Y94A 6312 Y15V,R35E 11182 N95W,S47V 16052 L84H,K118S 20922
R37F,N95A 6313 Y15V,R35F 11183 N95W,A48G 16053 L84H,K118M 20923
R37F,N95V 6314 Y15V,R35G 11184 N95W,E53A 16054 L84H,K118L 20924
R37F,T96A 6315 Y15V,R35I 11185 N95W,Y55A 16055 L84H,P121A 20925
R37F,K100E 6316 Y15V,R35L 11186 N95W,Y55F 16056 L84H,R122A 20926
R37F,K101E 6317 Y15V,R35M 11187 N95W,K57A 16057 L84H,R122E 20927
R37F,H102A 6318 Y15V,R35P 11188 N95W,K57E 16058 L84H,H124A 20928
R37F,K105A 6319 Y15V,R35S 11189 N95W,E60A 16059 L84H,Q127A 20929
R37F,K105E 6320 Y15V,R35V 11190 N95W,E60K 16060 L84H,Q127K 20930
R37F,K113A 6321 Y15V,D36A 11191 N95W,T61A 16061 L84H,K128E 20931
R37F,S116W 6322 Y15V,D36K 11192 N95W,Q63A 16062 L84H,L133A 20932
R37F,S116Y 6323 Y15V,R37E 11193 N95W,Y64A 16063 L84H,L133V 20933
R37F,S116F 6324 Y15V,R37A 11194 N95W,D68A 16064 L84I,K10E 20934
R37F,S116G 6325 Y15V,S38A 11195 N95W,T78A 16065 L84I,Y15A 20935
R37F,S116N 6326 Y15V,D39A 11196 N95W,P79A 16066 L84I,Y15F 20936
R37F,S116K 6327 Y15V,Q40A 11197 N95W,E81K 16067 L84I,S17A 20937
R37F,S116H 6328 Y15V,Q43A 11198 N95W,L84A 16068 L84I,S17K 20938
R37F,S116Q 6329 Y15V,Q43E 11199 N95W,E87K 16069 L84I,N18R 20939
R37F,C117K 6330 Y15V,Q43K 11200 N95W,E87Q 16070 L84I,H21A 20940
R37F,C117R 6331 Y15V,S47A 11201 N95W,E87V 16071 L84I,L26A 20941
R37F,C117S 6332 Y15V,S47V 11202 N95W,E87D 16072 L84I,P27A 20942
R37F,C117Q 6333 Y15V,A48G 11203 N95W,R88L 16073 L84I,D28K 20943
R37F,C117T 6334 Y15V,E53A 11204 N95W,R88Y 16074 L84I,T30A 20944
R37F,C117P 6335 Y15V,Y55A 11205 N95W,R88D 16075 L84I,T34A 20945
R37F,C117A 6336 Y15V,Y55F 11206 N95W,L89A 16076 L84I,R35A 20946
R37F,C117I 6337 Y15V,K57A 11207 N95W,E90K 16077 L84I,R35E 20947
R37F,K118W 6338 Y15V,K57E 11208 N95W,E91A 16078 L84I,R35F 20948
Figure imgf000165_0001
L
N95W,P121
R37G,Y15A 6371 Y15V,S116W 11241 16111 L84I,P79A 20981
A
N95W,R122
R37G,Y15F 6372 Y15V,S116Y 11242 16112 L84I,E81K 20982
A
N95W,R122
R37G,S17A 6373 Y15V,S116F 11243 16113 L84I,E87K 20983
E
N95W,H124
R37G,S17K 6374 Y15V,S116G 11244 16114 L84I,E87Q 20984
A
N95W,Q127
R37G,N18R 6375 Y15V,S116N 11245 16115 L84I,E87V 20985
A
N95W,Q127
R37G,H21A 6376 Y15V,S116K 11246 16116 L84I,E87D 20986
K
N95W,K128
R37G,L26A 6377 Y15V,S116H 11247 16117 L84I,R88L 20987
E
N95W,L133
R37G,P27A 6378 Y15V,S116Q 11248 16118 L84I,R88Y 20988
A
N95W,L133
R37G,D28K 6379 Y15V,C117K 11249 16119 L84I,R88D 20989
V
R37G,T30A 6380 Y15V,C117R 11250 N95Y,K10E 16120 L84I,L89A 20990
R37G,T34A 6381 Y15V,C117S 11251 N95Y,Y15A 16121 L84I,E90K 20991
R37G,R35A 6382 Y15V,C117Q 11252 N95Y,Y15F 16122 L84I,E91A 20992
R37G,R35E 6383 Y15V,C117T 11253 N95Y,S17A 16123 L84I,N92A 20993
R37G,R35F 6384 Y15V,C117P 11254 N95Y,S17K 16124 L84I,Y94V 20994
R37G,R35G 6385 Y15V,C117A 11255 N95Y,N18R 16125 L84I,Y94A 20995
R37G,R35I 6386 Y15V,C117I 11256 N95Y,H21A 16126 L84I,N95A 20996
R37G,R35L 6387 Y15V,K118W 11257 N95Y,L26A 16127 L84I,N95V 20997
R37G,R35M 6388 Y15V,K118H 11258 N95Y,P27A 16128 L84I,T96A 20998
R37G,R35P 6389 Y15V,K118R 11259 N95Y,D28K 16129 L84I,K100E 20999
R37G,R35S 6390 Y15V,K118S 11260 N95Y,T30A 16130 L84I,K101E 21000
R37G,R35V 6391 Y15V,K118M 11261 N95Y,T34A 16131 L84I,H102A 21001
R37G,D36A 6392 Y15V,K118L 11262 N95Y,R35A 16132 L84I,K105A 21002
R37G,D36K 6393 Y15V,P121A 11263 N95Y,R35E 16133 L84I,K105E 21003
R37G,S38A 6394 Y15V,R122A 11264 N95Y,R35F 16134 L84I,K113A 21004
R37G,D39A 6395 Y15V,R122E 11265 N95Y,R35G 16135 L84I,S116W 21005
R37G,Q40A 6396 Y15V,H124A 11266 N95Y,R35I 16136 L84I,S116Y 21006
R37G,Q43A 6397 Y15V,Q127A 11267 N95Y,R35L 16137 L84I,S116F 21007
R37G,Q43E 6398 Y15V,Q127K 11268 N95Y,R35M 16138 L84I,S116G 21008
R37G,Q43K 6399 Y15V,K128E 11269 N95Y,R35P 16139 L84I,S116N 21009
R37G,S47A 6400 Y15V,L133A 11270 N95Y,R35S 16140 L84I,S116K 21010
R37G,S47V 6401 Y15V,L133V 11271 N95Y,R35V 16141 L84I,S116H 21011
R37G,A48G 6402 Y15W,K10E 11272 N95Y,D36A 16142 L84I,S116Q 21012
R37G,E53A 6403 Y15W,S17A 11273 N95Y,D36K 16143 L84I,C117K 21013
R37G,Y55A 6404 Y15W,S17K 11274 N95Y,R37E 16144 L84I,C117R 21014
R37G,Y55F 6405 Y15W,N18R 11275 N95Y,R37A 16145 L84I,C117S 21015
R37G,K57A 6406 Y15W,H21A 11276 N95Y,S38A 16146 L84I,C117Q 21016
R37G,K57E 6407 Y15W,L26A 11277 N95Y,D39A 16147 L84I,C117T 21017
R37G,E60A 6408 Y15W,P27A 11278 N95Y,Q40A 16148 L84I,C117P 21018
R37G,E60K 6409 Y15W,D28K 11279 N95Y,Q43A 16149 L84I,C117A 21019
R37G,T61A 6410 Y15W,T30A 11280 N95Y,Q43E 16150 L84I,C117I 21020 R37G,Q63A 6411 Y15W,T34A 11281 N95Y,Q43K 16151 L84I,K118W 21021
R37G,Y64A 6412 Y15W,R35A 11282 N95Y,S47A 16152 L84I,K118H 21022
R37G,D68A 6413 Y15W,R35E 11283 N95Y,S47V 16153 L84I,K118R 21023
R37G,T78A 6414 Y15W,R35F 11284 N95Y,A48G 16154 L84I,K118S 21024
R37G,P79A 6415 Y15W,R35G 11285 N95Y,E53A 16155 L84I,K118M 21025
R37G,E81K 6416 Y15W,R35I 11286 N95Y,Y55A 16156 L84I,K118L 21026
R37G,L84A 6417 Y15W,R35L 11287 N95Y,Y55F 16157 L84I,P121A 21027
R37G,E87K 6418 Y15W,R35M 11288 N95Y,K57A 16158 L84I,R122A 21028
R37G,E87Q 6419 Y15W,R35P 11289 N95Y,K57E 16159 L84I,R122E 21029
R37G,E87V 6420 Y15W,R35S 11290 N95Y,E60A 16160 L84I,H124A 21030
R37G,E87D 6421 Y15W,R35V 11291 N95Y,E60K 16161 L84I,Q127A 21031
R37G,R88L 6422 Y15W,D36A 11292 N95Y,T61A 16162 L84I,Q127K 21032
R37G,R88Y 6423 Y15W,D36K 11293 N95Y,Q63A 16163 L84I,K128E 21033
R37G,R88D 6424 Y15W,R37E 11294 N95Y,Y64A 16164 L84I,L133A 21034
R37G,L89A 6425 Y15W,R37A 11295 N95Y,D68A 16165 L84I,L133V 21035
R37G,E90K 6426 Y15W,S38A 11296 N95Y,T78A 16166 L84K,K10E 21036
R37G,E91A 6427 Y15W,D39A 11297 N95Y,P79A 16167 L84K,Y15A 21037
R37G,N92A 6428 Y15W,Q40A 11298 N95Y,E81K 16168 L84K,Y15F 21038
R37G,Y94V 6429 Y15W,Q43A 11299 N95Y,L84A 16169 L84K,S17A 21039
R37G,Y94A 6430 Y15W,Q43E 11300 N95Y,E87K 16170 L84K,S17K 21040
R37G,N95A 6431 Y15W,Q43K 11301 N95Y,E87Q 16171 L84K,N18R 21041
R37G,N95V 6432 Y15W,S47A 11302 N95Y,E87V 16172 L84K,H21A 21042
R37G,T96A 6433 Y15W,S47V 11303 N95Y,E87D 16173 L84K,L26A 21043
R37G,K100E 6434 Y15W,A48G 11304 N95Y,R88L 16174 L84K,P27A 21044
R37G,K101E 6435 Y15W,E53A 11305 N95Y,R88Y 16175 L84K,D28K 21045
R37G,H102A 6436 Y15W,Y55A 11306 N95Y,R88D 16176 L84K,T30A 21046
R37G,K105A 6437 Y15W,Y55F 11307 N95Y,L89A 16177 L84K,T34A 21047
R37G,K105E 6438 Y15W,K57A 11308 N95Y,E90K 16178 L84K,R35A 21048
R37G,K113A 6439 Y15W,K57E 11309 N95Y,E91A 16179 L84K,R35E 21049
R37G,S116W 6440 Y15W,E60A 11310 N95Y,N92A 16180 L84K,R35F 21050
R37G,S116Y 6441 Y15W,E60K 11311 N95Y,Y94V 16181 L84K,R35G 21051
R37G,S116F 6442 Y15W,T61A 11312 N95Y,Y94A 16182 L84K,R35I 21052
R37G,S116G 6443 Y15W,Q63A 11313 N95Y,T96A 16183 L84K,R35L 21053
R37G,S116N 6444 Y15W,Y64A 11314 N95Y,K100E 16184 L84K,R35M 21054
R37G,S116K 6445 Y15W,D68A 11315 N95Y,K101E 16185 L84K,R35P 21055
N95Y,H102
R37G,S116H 6446 Y15W,T78A 11316 16186 L84K,R35S 21056
A
N95Y,K105
R37G,S116Q 6447 Y15W,P79A 11317 16187 L84K,R35V 21057
A
R37G,C117K 6448 Y15W,E81K 11318 N95Y,K105E 16188 L84K,D36A 21058
N95Y,K113
R37G,C117R 6449 Y15W,L84A 11319 16189 L84K,D36K 21059
A
N95Y,S116
R37G,C117S 6450 Y15W,E87K 11320 16190 L84K,R37E 21060
W
R37G,C117Q 6451 Y15W,E87Q 11321 N95Y,S116Y 16191 L84K,R37A 21061
R37G,C117T 6452 Y15W,E87V 11322 N95Y,S116F 16192 L84K,S38A 21062
R37G,C117P 6453 Y15W,E87D 11323 N95Y,S116G 16193 L84K,D39A 21063
R37G,C117A 6454 Y15W,R88L 11324 N95Y,S116N 16194 L84K,Q40A 21064 R37G,C117I 6455 Y15W,R88Y 11325 N95Y,S116K 16195 L84K,Q43A 21065
R37G,K118
6456 Y15W,R88D 11326 N95Y,S116H 16196 L84K,Q43E 21066 W
R37G,K118H 6457 Y15W,L89A 11327 N95Y,S116Q 16197 L84K,Q43K 21067
R37G,K118R 6458 Y15W,E90K 11328 N95Y,C117K 16198 L84K,S47A 21068
R37G,K118S 6459 Y15W,E91A 11329 N95Y,C117R 16199 L84K,S47V 21069
R37G,K118
6460 Y15W,N92A 11330 N95Y,C117S 16200 L84K,A48G 21070 M
R37G,K118L 6461 Y15W,Y94V 11331 N95Y,C117Q 16201 L84K,E53A 21071
R37G,P121A 6462 Y15W,Y94A 11332 N95Y,C117T 16202 L84K,Y55A 21072
R37G,R122A 6463 Y15W,N95A 11333 N95Y,C117P 16203 L84K,Y55F 21073
R37G,R122E 6464 Y15W,N95V 11334 N95Y,C117A 16204 L84K,K57A 21074
R37G,H124A 6465 Y15W,T96A 11335 N95Y,C117I 16205 L84K,K57E 21075
N95Y,K118
R37G,Q127A 6466 Y15W,K100E 11336 16206 L84K,E60A 21076
W
N95Y,K118
R37G,Q127K 6467 Y15W,K101E 11337 16207 L84K,E60K 21077
H
R37G,K128E 6468 Y15W,H102A 11338 N95Y,K118R 16208 L84K,T61A 21078
R37G,L133A 6469 Y15W,K105A 11339 N95Y,K118S 16209 L84K,Q63A 21079
N95Y,K118
R37G,L133V 6470 Y15W,K105E 11340 16210 L84K,Y64A 21080
M
R37H,Q43D 6471 Y15W,K113A 11341 N95Y,K118L 16211 L84K,D68A 21081
R37H,Q43F 6472 Y15W,S116W 11342 N95Y,P121A 16212 L84K,T78A 21082
R37H,Q43G 6473 Y15W,S116Y 11343 N95Y,R122A 16213 L84K,P79A 21083
R37H,Q43H 6474 Y15W,S116F 11344 N95Y,R122E 16214 L84K,E81K 21084
N95Y,H124
R37H,Q43I 6475 Y15W,S116G 11345 16215 L84K,E87K 21085
A
N95Y,Q127
R37H,Q43L 6476 Y15W,S116N 11346 16216 L84K,E87Q 21086
A
N95Y,Q127
R37H,Q43M 6477 Y15W,S116K 11347 16217 L84K,E87V 21087
K
R37H,Q43N 6478 Y15W,S116H 11348 N95Y,K128E 16218 L84K,E87D 21088
R37H,Q43P 6479 Y15W,S116Q 11349 N95Y,L133A 16219 L84K,R88L 21089
R37H,Q43S 6480 Y15W,C117K 11350 N95Y,L133V 16220 L84K,R88Y 21090
R37H,Q43T 6481 Y15W,C117R 11351 D36E,K10E 16221 L84K,R88D 21091
R37H,Q43V 6482 Y15W,C117S 11352 D36E,Y15A 16222 L84K,L89A 21092
R37H,Q43W 6483 Y15W,C117Q 11353 D36E,Y15F 16223 L84K,E90K 21093
R37H,Q43Y 6484 Y15W,C117T 11354 D36E,S17A 16224 L84K,E91A 21094
R37H,D28A 6485 Y15W,C117P 11355 D36E,S17K 16225 L84K,N92A 21095
R37H,E49A 6486 Y15W,C117A 11356 D36E,N18R 16226 L84K,Y94V 21096
R37H,P121A 6487 Y15W,C117I 11357 D36E,H21A 16227 L84K,Y94A 21097
Y15W,K118
R37H,K10E 6488 11358 D36E,L26A 16228 L84K,N95A 21098
W
R37H,Y15A 6489 Y15W,K118H 11359 D36E,P27A 16229 L84K,N95V 21099
R37H,Y15F 6490 Y15W,K118R 11360 D36E,D28K 16230 L84K,T96A 21100
R37H,S17A 6491 Y15W,K118S 11361 D36E,T30A 16231 L84K,K100E 21101
R37H,S17K 6492 Y15W,K118M 11362 D36E,T34A 16232 L84K,K101E 21102
R37H,N18R 6493 Y15W,K118L 11363 D36E,R35A 16233 L84K,H102A 21103
R37H,H21A 6494 Y15W,P121A 11364 D36E,R35E 16234 L84K,K105A 21104
R37H,L26A 6495 Y15W,R122A 11365 D36E,R35F 16235 L84K,K105E 21105 R37H,P27A 6496 Y15W,R122E 11366 D36E,R35G 16236 L84K,K113A 21106
R37H,D28K 6497 Y15W,H124A 11367 D36E,R35I 16237 L84K,S116W 21107
R37H,T30A 6498 Y15W,Q127A 11368 D36E,R35L 16238 L84K,S116Y 21108
R37H,T34A 6499 Y15W,Q127K 11369 D36E,R35M 16239 L84K,S116F 21109
R37H,R35A 6500 Y15W,K128E 11370 D36E,R35P 16240 L84K,S116G 21110
R37H,R35E 6501 Y15W,L133A 11371 D36E,R35S 16241 L84K,S116N 21111
R37H,R35F 6502 Y15W,L133V 11372 D36E,R35V 16242 L84K,S116K 21112
R37H,R35G 6503 L133D,K10E 11373 D36E,R37E 16243 L84K,S116H 21113
R37H,R35I 6504 L133D,Y15A 11374 D36E,R37A 16244 L84K,S116Q 21114
R37H,R35L 6505 L133D,Y15F 11375 D36E,S38A 16245 L84K,C117K 21115
R37H,R35M 6506 L133D,S17A 11376 D36E,D39A 16246 L84K,C117R 21116
R37H,R35P 6507 L133D,S17K 11377 D36E,Q40A 16247 L84K,C117S 21117
R37H,R35S 6508 L133D,N18R 11378 D36E,Q43A 16248 L84K,C117Q 21118
R37H,R35V 6509 L133D,H21A 11379 D36E,Q43E 16249 L84K,C117T 21119
R37H,D36A 6510 L133D,L26A 11380 D36E,Q43K 16250 L84K,C117P 21120
R37H,D36K 6511 L133D,P27A 11381 D36E,S47A 16251 L84K,C117A 21121
R37H,S38A 6512 L133D,D28K 11382 D36E,S47V 16252 L84K,C117I 21122
R37H,D39A 6513 L133D,T30A 11383 D36E,A48G 16253 L84K,K118W 21123
R37H,Q40A 6514 L133D,T34A 11384 D36E,E53A 16254 L84K,K118H 21124
R37H,Q43A 6515 L133D,R35A 11385 D36E,Y55A 16255 L84K,K118R 21125
R37H,Q43E 6516 L133D,R35E 11386 D36E,Y55F 16256 L84K,K118S 21126
R37H,Q43K 6517 L133D,R35F 11387 D36E,K57A 16257 L84K,K118M 21127
R37H,S47A 6518 L133D,R35G 11388 D36E,K57E 16258 L84K,K118L 21128
R37H,S47V 6519 L133D,R35I 11389 D36E,E60A 16259 L84K,P121A 21129
R37H,A48G 6520 L133D,R35L 11390 D36E,E60K 16260 L84K,R122A 21130
R37H,E53A 6521 L133D,R35M 11391 D36E,T61A 16261 L84K,R122E 21131
R37H,Y55A 6522 L133D,R35P 11392 D36E,Q63A 16262 L84K,H124A 21132
R37H,Y55F 6523 L133D,R35S 11393 D36E,Y64A 16263 L84K,Q127A 21133
R37H,K57A 6524 L133D,R35V 11394 D36E,D68A 16264 L84K,Q127K 21134
R37H,K57E 6525 L133D,D36A 11395 D36E,T78A 16265 L84K,K128E 21135
R37H,E60A 6526 L133D,D36K 11396 D36E,P79A 16266 L84K,L133A 21136
R37H,E60K 6527 L133D,R37E 11397 D36E,E81K 16267 L84K,L133V 21137
R37H,T61A 6528 L133D,R37A 11398 D36E,L84A 16268 L84M,K10E 21138
R37H,Q63A 6529 L133D,S38A 11399 D36E,E87K 16269 L84M,Y15A 21139
R37H,Y64A 6530 L133D,D39A 11400 D36E,E87Q 16270 L84M,Y15F 21140
R37H,D68A 6531 L133D,Q40A 11401 D36E,E87V 16271 L84M,S17A 21141
R37H,T78A 6532 L133D,Q43A 11402 D36E,E87D 16272 L84M,S17K 21142
R37H,P79A 6533 L133D,Q43E 11403 D36E,R88L 16273 L84M,N18R 21143
R37H,E81K 6534 L133D,Q43K 11404 D36E,R88Y 16274 L84M,H21A 21144
R37H,L84A 6535 L133D,S47A 11405 D36E,R88D 16275 L84M,L26A 21145
R37H,E87K 6536 L133D,S47V 11406 D36E,L89A 16276 L84M,P27A 21146
R37H,E87Q 6537 L133D,A48G 11407 D36E,E90K 16277 L84M,D28K 21147
R37H,E87V 6538 L133D,E53A 11408 D36E,E91A 16278 L84M,T30A 21148
R37H,E87D 6539 L133D,Y55A 11409 D36E,N92A 16279 L84M,T34A 21149
R37H,R88L 6540 L133D,Y55F 11410 D36E,Y94V 16280 L84M,R35A 21150
R37H,R88Y 6541 L133D,K57A 11411 D36E,Y94A 16281 L84M,R35E 21151 R37H,R88D 6542 L133D,K57E 11412 D36E,N95A 16282 L84M,R35F 21152
R37H,L89A 6543 L133D,E60A 11413 D36E,N95V 16283 L84M,R35G 21153
R37H,E90K 6544 L133D,E60K 11414 D36E,T96A 16284 L84M,R35I 21154
R37H,E91A 6545 L133D,T61A 11415 D36E,K100E 16285 L84M,R35L 21155
R37H,N92A 6546 L133D,Q63A 11416 D36E,K101E 16286 L84M,R35M 21156
R37H,Y94V 6547 L133D,Y64A 11417 D36E,H102A 16287 L84M,R35P 21157
R37H,Y94A 6548 L133D,D68A 11418 D36E,K105A 16288 L84M,R35S 21158
R37H,N95A 6549 L133D,T78A 11419 D36E,K105E 16289 L84M,R35V 21159
R37H,N95V 6550 L133D,P79A 11420 D36E,K113A 16290 L84M,D36A 21160
R37H,T96A 6551 L133D,E81K 11421 D36E,S116W 16291 L84M,D36K 21161
R37H,K100E 6552 L133D,L84A 11422 D36E,S116Y 16292 L84M,R37E 21162
R37H,K101E 6553 L133D,E87K 11423 D36E,S116F 16293 L84M,R37A 21163
R37H,H102A 6554 L133D,E87Q 11424 D36E,S116G 16294 L84M,S38A 21164
R37H,K105A 6555 L133D,E87V 11425 D36E,S116N 16295 L84M,D39A 21165
R37H,K105E 6556 L133D,E87D 11426 D36E,S116K 16296 L84M,Q40A 21166
R37H,K113A 6557 L133D,R88L 11427 D36E,S116H 16297 L84M,Q43A 21167
R37H,S116W 6558 L133D,R88Y 11428 D36E,S116Q 16298 L84M,Q43E 21168
R37H,S116Y 6559 L133D,R88D 11429 D36E,C117K 16299 L84M,Q43K 21169
R37H,S116F 6560 L133D,L89A 11430 D36E,C117R 16300 L84M,S47A 21170
R37H,S116G 6561 L133D,E90K 11431 D36E,C117S 16301 L84M,S47V 21171
R37H,S116N 6562 L133D,E91A 11432 D36E,C117Q 16302 L84M,A48G 21172
R37H,S116K 6563 L133D,N92A 11433 D36E,C117T 16303 L84M,E53A 21173
R37H,S116H 6564 L133D,Y94V 11434 D36E,C117P 16304 L84M,Y55A 21174
R37H,S116Q 6565 L133D,Y94A 11435 D36E,C117A 16305 L84M,Y55F 21175
R37H,C117K 6566 L133D,N95A 11436 D36E,C117I 16306 L84M,K57A 21176
D36E,K118
R37H,C117R 6567 L133D,N95V 11437 16307 L84M,K57E 21177
W
R37H,C117S 6568 L133D,T96A 11438 D36E,K118H 16308 L84M,E60A 21178
R37H,C117Q 6569 L133D,K100E 11439 D36E,K118R 16309 L84M,E60K 21179
R37H,C117T 6570 L133D,K101E 11440 D36E,K118S 16310 L84M,T61A 21180
D36E,K118
R37H,C117P 6571 L133D,H102A 11441 16311 L84M,Q63A 21181
M
R37H,C117A 6572 L133D,K105A 11442 D36E,K118L 16312 L84M,Y64A 21182
R37H,C117I 6573 L133D,K105E 11443 D36E,P121A 16313 L84M,D68A 21183
R37H,K118
6574 L133D,K113A 11444 D36E,R122A 16314 L84M,T78A 21184 W
L133D,S116
R37H,K118H 6575 11445 D36E,R122E 16315 L84M,P79A 21185
W
R37H,K118R 6576 L133D,S116Y 11446 D36E,H124A 16316 L84M,E81K 21186
R37H,K118S 6577 L133D,S116F 11447 D36E,Q127A 16317 L84M,E87K 21187
R37H,K118
6578 L133D,S116G 11448 D36E,Q127K 16318 L84M,E87Q 21188 M
R37H,K118L 6579 L133D,S116N 11449 D36E,K128E 16319 L84M,E87V 21189
R37H,P121A 6580 L133D,S116K 11450 D36E,L133A 16320 L84M,E87D 21190
R37H,R122A 6581 L133D,S116H 11451 D36E,L133V 16321 L84M,R88L 21191
R37H,R122E 6582 L133D,S116Q 11452 D36F,K10E 16322 L84M,R88Y 21192
R37H,H124A 6583 L133D,C117K 11453 D36F,Y15A 16323 L84M,R88D 21193
R37H,Q127A 6584 L133D,C117R 11454 D36F,Y15F 16324 L84M,L89A 21194 R37H,Q127K 6585 L133D,C117S 11455 D36F,S17A 16325 L84M,E90K 21195
R37H,K128E 6586 L133D,C117Q 11456 D36F,S17K 16326 L84M,E91A 21196
R37H,L133A 6587 L133D,C117T 11457 D36F,N18R 16327 L84M,N92A 21197
R37H,L133V 6588 L133D,C117P 11458 D36F,H21A 16328 L84M,Y94V 21198
R37I,Q43D 6589 L133D,C117A 11459 D36F,L26A 16329 L84M,Y94A 21199
R37I,Q43F 6590 L133D,C117I 11460 D36F,P27A 16330 L84M,N95A 21200
L133D,K118
R37I,Q43G 6591 11461 D36F,D28K 16331 L84M,N95V 21201
W
R37I,Q43H 6592 L133D,K118H 11462 D36F,T30A 16332 L84M,T96A 21202
R37I,Q43I 6593 L133D,K118R 11463 D36F,T34A 16333 L84M,K100E 21203
R37I,Q43L 6594 L133D,K118S 11464 D36F,R35A 16334 L84M,K101E 21204
L133D,K118
R37I,Q43M 6595 11465 D36F,R35E 16335 L84M,H102A 21205
M
R37I,Q43N 6596 L133D,K118L 11466 D36F,R35F 16336 L84M,K105A 21206
R37I,Q43P 6597 L133D,P121A 11467 D36F,R35G 16337 L84M,K105E 21207
R37I,Q43S 6598 L133D,R122A 11468 D36F,R35I 16338 L84M,K113A 21208
R37I,Q43T 6599 L133D,R122E 11469 D36F,R35L 16339 L84M,S116W 21209
R37I,Q43V 6600 L133D,H124A 11470 D36F,R35M 16340 L84M,S116Y 21210
R37I,Q43W 6601 L133D,Q127A 11471 D36F,R35P 16341 L84M,S116F 21211
R37I,Q43Y 6602 L133D,Q127K 11472 D36F,R35S 16342 L84M,S116G 21212
R37I,D28A 6603 L133D,K128E 11473 D36F,R35V 16343 L84M,S116N 21213
R37I,E49A 6604 L133E,K10E 11474 D36F,R37E 16344 L84M,S116K 21214
R37I,P121A 6605 L133E,Y15A 11475 D36F,R37A 16345 L84M,S116H 21215
R37I,K10E 6606 L133E,Y15F 11476 D36F,S38A 16346 L84M,S116Q 21216
R37I,Y15A 6607 L133E,S17A 11477 D36F,D39A 16347 L84M,C117K 21217
R37I,Y15F 6608 L133E,S17K 11478 D36F,Q40A 16348 L84M,C117R 21218
R37I,S17A 6609 L133E,N18R 11479 D36F,Q43A 16349 L84M,C117S 21219
R37I,S17K 6610 L133E,H21A 11480 D36F,Q43E 16350 L84M,C117Q 21220
R37I,N18R 6611 L133E,L26A 11481 D36F,Q43K 16351 L84M,C117T 21221
R37I,H21A 6612 L133E,P27A 11482 D36F,S47A 16352 L84M,C117P 21222
R37I,L26A 6613 L133E,D28K 11483 D36F,S47V 16353 L84M,C117A 21223
R37I,P27A 6614 L133E,T30A 11484 D36F,A48G 16354 L84M,C117I 21224
R37I,D28K 6615 L133E,T34A 11485 D36F,E53A 16355 L84M,K118W 21225
R37I,T30A 6616 L133E,R35A 11486 D36F,Y55A 16356 L84M,K118H 21226
R37I,T34A 6617 L133E,R35E 11487 D36F,Y55F 16357 L84M,K118R 21227
R37I,R35A 6618 L133E,R35F 11488 D36F,K57A 16358 L84M,K118S 21228
R37I,R35E 6619 L133E,R35G 11489 D36F,K57E 16359 L84M,K118M 21229
R37I,R35F 6620 L133E,R35I 11490 D36F,E60A 16360 L84M,K118L 21230
R37I,R35G 6621 L133E,R35L 11491 D36F,E60K 16361 L84M,P121A 21231
R37I,R35I 6622 L133E,R35M 11492 D36F,T61A 16362 L84M,R122A 21232
R37I,R35L 6623 L133E,R35P 11493 D36F,Q63A 16363 L84M,R122E 21233
R37I,R35M 6624 L133E,R35S 11494 D36F,Y64A 16364 L84M,H124A 21234
R37I,R35P 6625 L133E,R35V 11495 D36F,D68A 16365 L84M,Q127A 21235
R37I,R35S 6626 L133E,D36A 11496 D36F,T78A 16366 L84M,Q127K 21236
R37I,R35V 6627 L133E,D36K 11497 D36F,P79A 16367 L84M,K128E 21237
R37I,D36A 6628 L133E,R37E 11498 D36F,E81K 16368 L84M,L133A 21238
R37I,D36K 6629 L133E,R37A 11499 D36F,L84A 16369 L84M,L133V 21239 R37I,S38A 6630 L133E,S38A 11500 D36F,E87K 16370 L84N,K10E 21240
R37I,D39A 6631 L133E,D39A 11501 D36F,E87Q 16371 L84N,Y15A 21241
R37I,Q40A 6632 L133E,Q40A 11502 D36F,E87V 16372 L84N,Y15F 21242
R37I,Q43A 6633 L133E,Q43A 11503 D36F,E87D 16373 L84N,S17A 21243
R37I,Q43E 6634 L133E,Q43E 11504 D36F,R88L 16374 L84N,S17K 21244
R37I,Q43K 6635 L133E,Q43K 11505 D36F,R88Y 16375 L84N,N18R 21245
R37I,S47A 6636 L133E,S47A 11506 D36F,R88D 16376 L84N,H21A 21246
R37I,S47V 6637 L133E,S47V 11507 D36F,L89A 16377 L84N,L26A 21247
R37I,A48G 6638 L133E,A48G 11508 D36F,E90K 16378 L84N,P27A 21248
R37I,E53A 6639 L133E,E53A 11509 D36F,E91A 16379 L84N,D28K 21249
R37I,Y55A 6640 L133E,Y55A 11510 D36F,N92A 16380 L84N,T30A 21250
R37I,Y55F 6641 L133E,Y55F 11511 D36F,Y94V 16381 L84N,T34A 21251
R37I,K57A 6642 L133E,K57A 11512 D36F,Y94A 16382 L84N,R35A 21252
R37I,K57E 6643 L133E,K57E 11513 D36F,N95A 16383 L84N,R35E 21253
R37I,E60A 6644 L133E,E60A 11514 D36F,N95V 16384 L84N,R35F 21254
R37I,E60K 6645 L133E,E60K 11515 D36F,T96A 16385 L84N,R35G 21255
R37I,T61A 6646 L133E,T61A 11516 D36F,K100E 16386 L84N,R35I 21256
R37I,Q63A 6647 L133E,Q63A 11517 D36F,K101E 16387 L84N,R35L 21257
R37I,Y64A 6648 L133E,Y64A 11518 D36F,H102A 16388 L84N,R35M 21258
R37I,D68A 6649 L133E,D68A 11519 D36F,K105A 16389 L84N,R35P 21259
R37I,T78A 6650 L133E,T78A 11520 D36F,K105E 16390 L84N,R35S 21260
R37I,P79A 6651 L133E,P79A 11521 D36F,K113A 16391 L84N,R35V 21261
R37I,E81K 6652 L133E,E81K 11522 D36F,S116W 16392 L84N,D36A 21262
R37I,L84A 6653 L133E,L84A 11523 D36F,S116Y 16393 L84N,D36K 21263
R37I,E87K 6654 L133E,E87K 11524 D36F,S116F 16394 L84N,R37E 21264
R37I,E87Q 6655 L133E,E87Q 11525 D36F,S116G 16395 L84N,R37A 21265
R37I,E87V 6656 L133E,E87V 11526 D36F,S116N 16396 L84N,S38A 21266
R37I,E87D 6657 L133E,E87D 11527 D36F,S116K 16397 L84N,D39A 21267
R37I,R88L 6658 L133E,R88L 11528 D36F,S116H 16398 L84N,Q40A 21268
R37I,R88Y 6659 L133E,R88Y 11529 D36F,S116Q 16399 L84N,Q43A 21269
R37I,R88D 6660 L133E,R88D 11530 D36F,C117K 16400 L84N,Q43E 21270
R37I,L89A 6661 L133E,L89A 11531 D36F,C117R 16401 L84N,Q43K 21271
R37I,E90K 6662 L133E,E90K 11532 D36F,C117S 16402 L84N,S47A 21272
R37I,E91A 6663 L133E,E91A 11533 D36F,C117Q 16403 L84N,S47V 21273
R37I,N92A 6664 L133E,N92A 11534 D36F,C117T 16404 L84N,A48G 21274
R37I,Y94V 6665 L133E,Y94V 11535 D36F,C117P 16405 L84N,E53A 21275
R37I,Y94A 6666 L133E,Y94A 11536 D36F,C117A 16406 L84N,Y55A 21276
R37I,N95A 6667 L133E,N95A 11537 D36F,C117I 16407 L84N,Y55F 21277
D36F,K118
R37I,N95V 6668 L133E,N95V 11538 16408 L84N,K57A 21278
W
R37I,T96A 6669 L133E,T96A 11539 D36F,K118H 16409 L84N,K57E 21279
R37I,K100E 6670 L133E,K100E 11540 D36F,K118R 16410 L84N,E60A 21280
R37I,K101E 6671 L133E,K101E 11541 D36F,K118S 16411 L84N,E60K 21281
D36F,K118
R37I,H102A 6672 L133E,H102A 11542 16412 L84N,T61A 21282
M
R37I,K105A 6673 L133E,K105A 11543 D36F,K118L 16413 L84N,Q63A 21283
R37I,K105E 6674 L133E,K105E 11544 D36F,P121A 16414 L84N,Y64A 21284 R37I,K113A 6675 L133E,K113A 11545 D36F,R122A 16415 L84N,D68A 21285
R37I,S116W 6676 L133E,S116W 11546 D36F,R122E 16416 L84N,T78A 21286
R37I,S116Y 6677 L133E,S116Y 11547 D36F,H124A 16417 L84N,P79A 21287
R37I,S116F 6678 L133E,S116F 11548 D36F,Q127A 16418 L84N,E81K 21288
R37I,S116G 6679 L133E,S116G 11549 D36F,Q127K 16419 L84N,E87K 21289
R37I,S116N 6680 L133E,S116N 11550 D36F,K128E 16420 L84N,E87Q 21290
R37I,S116K 6681 L133E,S116K 11551 D36F,L133A 16421 L84N,E87V 21291
R37I,S116H 6682 L133E,S116H 11552 D36F,L133V 16422 L84N,E87D 21292
R37I,S116Q 6683 L133E,S116Q 11553 D36G,K10E 16423 L84N,R88L 21293
R37I,C117K 6684 L133E,C117K 11554 D36G,Y15A 16424 L84N,R88Y 21294
R37I,C117R 6685 L133E,C117R 11555 D36G,Y15F 16425 L84N,R88D 21295
R37I,C117S 6686 L133E,C117S 11556 D36G,S17A 16426 L84N,L89A 21296
R37I,C117Q 6687 L133E,C117Q 11557 D36G,S17K 16427 L84N,E90K 21297
R37I,C117T 6688 L133E,C117T 11558 D36G,N18R 16428 L84N,E91A 21298
R37I,C117P 6689 L133E,C117P 11559 D36G,H21A 16429 L84N,N92A 21299
R37I,C117A 6690 L133E,C117A 11560 D36G,L26A 16430 L84N,Y94V 21300
R37I,C117I 6691 L133E,C117I 11561 D36G,P27A 16431 L84N,Y94A 21301
L133E,K118
R37I,K118W 6692 11562 D36G,D28K 16432 L84N,N95A 21302
W
R37I,K118H 6693 L133E,K118H 11563 D36G,T30A 16433 L84N,N95V 21303
R37I,K118R 6694 L133E,K118R 11564 D36G,T34A 16434 L84N,T96A 21304
R37I,K118S 6695 L133E,K118S 11565 D36G,R35A 16435 L84N,K100E 21305
L133E,K118
R37I,K118M 6696 11566 D36G,R35E 16436 L84N,K101E 21306
M
R37I,K118L 6697 L133E,K118L 11567 D36G,R35F 16437 L84N,H102A 21307
R37I,P121A 6698 L133E,P121A 11568 D36G,R35G 16438 L84N,K105A 21308
R37I,R122A 6699 L133E,R122A 11569 D36G,R35I 16439 L84N,K105E 21309
R37I,R122E 6700 L133E,R122E 11570 D36G,R35L 16440 L84N,K113A 21310
R37I,H124A 6701 L133E,H124A 11571 D36G,R35M 16441 L84N,S116W 21311
R37I,Q127A 6702 L133E,Q127A 11572 D36G,R35P 16442 L84N,S116Y 21312
R37I,Q127K 6703 L133E,Q127K 11573 D36G,R35S 16443 L84N,S116F 21313
R37I,K128E 6704 L133E,K128E 11574 D36G,R35V 16444 L84N,S116G 21314
R37I,L133A 6705 L133F,K10E 11575 D36G,R37E 16445 L84N,S116N 21315
R37I,L133V 6706 L133F,Y15A 11576 D36G,R37A 16446 L84N,S116K 21316
R37K,Q43D 6707 L133F,Y15F 11577 D36G,S38A 16447 L84N,S116H 21317
R37K,Q43F 6708 L133F,S17A 11578 D36G,D39A 16448 L84N,S116Q 21318
R37K,Q43G 6709 L133F,S17K 11579 D36G,Q40A 16449 L84N,C117K 21319
R37K,Q43H 6710 L133F,N18R 11580 D36G,Q43A 16450 L84N,C117R 21320
R37K,Q43I 6711 L133F,H21A 11581 D36G,Q43E 16451 L84N,C117S 21321
R37K,Q43L 6712 L133F,L26A 11582 D36G,Q43K 16452 L84N,C117Q 21322
R37K,Q43M 6713 L133F,P27A 11583 D36G,S47A 16453 L84N,C117T 21323
R37K,Q43N 6714 L133F,D28K 11584 D36G,S47V 16454 L84N,C117P 21324
R37K,Q43P 6715 L133F,T30A 11585 D36G,A48G 16455 L84N,C117A 21325
R37K,Q43S 6716 L133F,T34A 11586 D36G,E53A 16456 L84N,C117I 21326
R37K,Q43T 6717 L133F,R35A 11587 D36G,Y55A 16457 L84N,K118W 21327
R37K,Q43V 6718 L133F,R35E 11588 D36G,Y55F 16458 L84N,K118H 21328
R37K,Q43W 6719 L133F,R35F 11589 D36G,K57A 16459 L84N,K118R 21329 R37K,Q43Y 6720 L133F,R35G 11590 D36G,K57E 16460 L84N,K118S 21330
R37K,D28A 6721 L133F,R35I 11591 D36G,E60A 16461 L84N,K118M 21331
R37K,E49A 6722 L133F,R35L 11592 D36G,E60K 16462 L84N,K118L 21332
R37K,P121A 6723 L133F,R35M 11593 D36G,T61A 16463 L84N,P121A 21333
R37K,K10E 6724 L133F,R35P 11594 D36G,Q63A 16464 L84N,R122A 21334
R37K,Y15A 6725 L133F,R35S 11595 D36G,Y64A 16465 L84N,R122E 21335
R37K,Y15F 6726 L133F,R35V 11596 D36G,D68A 16466 L84N,H124A 21336
R37K,S17A 6727 L133F,D36A 11597 D36G,T78A 16467 L84N,Q127A 21337
R37K,S17K 6728 L133F,D36K 11598 D36G,P79A 16468 L84N,Q127K 21338
R37K,N18R 6729 L133F,R37E 11599 D36G,E81K 16469 L84N,K128E 21339
R37K,H21A 6730 L133F,R37A 11600 D36G,L84A 16470 L84N,L133A 21340
R37K,L26A 6731 L133F,S38A 11601 D36G,E87K 16471 L84N,L133V 21341
R37K,P27A 6732 L133F,D39A 11602 D36G,E87Q 16472 L84P,K10E 21342
R37K,D28K 6733 L133F,Q40A 11603 D36G,E87V 16473 L84P,Y15A 21343
R37K,T30A 6734 L133F,Q43A 11604 D36G,E87D 16474 L84P,Y15F 21344
R37K,T34A 6735 L133F,Q43E 11605 D36G,R88L 16475 L84P,S17A 21345
R37K,R35A 6736 L133F,Q43K 11606 D36G,R88Y 16476 L84P,S17K 21346
R37K,R35E 6737 L133F,S47A 11607 D36G,R88D 16477 L84P,N18R 21347
R37K,R35F 6738 L133F,S47V 11608 D36G,L89A 16478 L84P,H21A 21348
R37K,R35G 6739 L133F,A48G 11609 D36G,E90K 16479 L84P,L26A 21349
R37K,R35I 6740 L133F,E53A 11610 D36G,E91A 16480 L84P,P27A 21350
R37K,R35L 6741 L133F,Y55A 11611 D36G,N92A 16481 L84P,D28K 21351
R37K,R35M 6742 L133F,Y55F 11612 D36G,Y94V 16482 L84P,T30A 21352
R37K,R35P 6743 L133F,K57A 11613 D36G,Y94A 16483 L84P,T34A 21353
R37K,R35S 6744 L133F,K57E 11614 D36G,N95A 16484 L84P,R35A 21354
R37K,R35V 6745 L133F,E60A 11615 D36G,N95V 16485 L84P,R35E 21355
R37K,D36A 6746 L133F,E60K 11616 D36G,T96A 16486 L84P,R35F 21356
R37K,D36K 6747 L133F,T61A 11617 D36G,K100E 16487 L84P,R35G 21357
R37K,S38A 6748 L133F,Q63A 11618 D36G,K101E 16488 L84P,R35I 21358
D36G,H102
R37K,D39A 6749 L133F,Y64A 11619 16489 L84P,R35L 21359
A
D36G,K105
R37K,Q40A 6750 L133F,D68A 11620 16490 L84P,R35M 21360
A
R37K,Q43A 6751 L133F,T78A 11621 D36G,K105E 16491 L84P,R35P 21361
D36G,K113
R37K,Q43E 6752 L133F,P79A 11622 16492 L84P,R35S 21362
A
D36G,S116
R37K,Q43K 6753 L133F,E81K 11623 16493 L84P,R35V 21363
W
R37K,S47A 6754 L133F,L84A 11624 D36G,S116Y 16494 L84P,D36A 21364
R37K,S47V 6755 L133F,E87K 11625 D36G,S116F 16495 L84P,D36K 21365
R37K,A48G 6756 L133F,E87Q 11626 D36G,S116G 16496 L84P,R37E 21366
R37K,E53A 6757 L133F,E87V 11627 D36G,S116N 16497 L84P,R37A 21367
R37K,Y55A 6758 L133F,E87D 11628 D36G,S116K 16498 L84P,S38A 21368
R37K,Y55F 6759 L133F,R88L 11629 D36G,S116H 16499 L84P,D39A 21369
R37K,K57A 6760 L133F,R88Y 11630 D36G,S116Q 16500 L84P,Q40A 21370
R37K,K57E 6761 L133F,R88D 11631 D36G,C117K 16501 L84P,Q43A 21371
R37K,E60A 6762 L133F,L89A 11632 D36G,C117R 16502 L84P,Q43E 21372
R37K,E60K 6763 L133F,E90K 11633 D36G,C117S 16503 L84P,Q43K 21373 R37K,T61A 6764 L133F,E91A 11634 D36G,C117Q 16504 L84P,S47A 21374
R37K,Q63A 6765 L133F,N92A 11635 D36G,C117T 16505 L84P,S47V 21375
R37K,Y64A 6766 L133F,Y94V 11636 D36G,C117P 16506 L84P,A48G 21376
R37K,D68A 6767 L133F,Y94A 11637 D36G,C117A 16507 L84P,E53A 21377
R37K,T78A 6768 L133F,N95A 11638 D36G,C117I 16508 L84P,Y55A 21378
D36G,K118
R37K,P79A 6769 L133F,N95V 11639 16509 L84P,Y55F 21379
W
D36G,K118
R37K,E81K 6770 L133F,T96A 11640 16510 L84P,K57A 21380
H
R37K,L84A 6771 L133F,K100E 11641 D36G,K118R 16511 L84P,K57E 21381
R37K,E87K 6772 L133F,K101E 11642 D36G,K118S 16512 L84P,E60A 21382
D36G,K118
R37K,E87Q 6773 L133F,H102A 11643 16513 L84P,E60K 21383
M
R37K,E87V 6774 L133F,K105A 11644 D36G,K118L 16514 L84P,T61A 21384
R37K,E87D 6775 L133F,K105E 11645 D36G,P121A 16515 L84P,Q63A 21385
R37K,R88L 6776 L133F,K113A 11646 D36G,R122A 16516 L84P,Y64A 21386
R37K,R88Y 6777 L133F,S116W 11647 D36G,R122E 16517 L84P,D68A 21387
D36G,H124
R37K,R88D 6778 L133F,S116Y 11648 16518 L84P,T78A 21388
A
D36G,Q127
R37K,L89A 6779 L133F,S116F 11649 16519 L84P,P79A 21389
A
D36G,Q127
R37K,E90K 6780 L133F,S116G 11650 16520 L84P,E81K 21390
K
R37K,E91A 6781 L133F,S116N 11651 D36G,K128E 16521 L84P,E87K 21391
R37K,N92A 6782 L133F,S116K 11652 D36G,L133A 16522 L84P,E87Q 21392
R37K,Y94V 6783 L133F,S116H 11653 D36G,L133V 16523 L84P,E87V 21393
R37K,Y94A 6784 L133F,S116Q 11654 D36H,K10E 16524 L84P,E87D 21394
R37K,N95A 6785 L133F,C117K 11655 D36H,Y15A 16525 L84P,R88L 21395
R37K,N95V 6786 L133F,C117R 11656 D36H,Y15F 16526 L84P,R88Y 21396
R37K,T96A 6787 L133F,C117S 11657 D36H,S17A 16527 L84P,R88D 21397
R37K,K100E 6788 L133F,C117Q 11658 D36H,S17K 16528 L84P,L89A 21398
R37K,K101E 6789 L133F,C117T 11659 D36H,N18R 16529 L84P,E90K 21399
R37K,H102A 6790 L133F,C117P 11660 D36H,H21A 16530 L84P,E91A 21400
R37K,K105A 6791 L133F,C117A 11661 D36H,L26A 16531 L84P,N92A 21401
R37K,K105E 6792 L133F,C117I 11662 D36H,P27A 16532 L84P,Y94V 21402
L133F,K118
R37K,K113A 6793 11663 D36H,D28K 16533 L84P,Y94A 21403
W
R37K,S116W 6794 L133F,K118H 11664 D36H,T30A 16534 L84P,N95A 21404
R37K,S116Y 6795 L133F,K118R 11665 D36H,T34A 16535 L84P,N95V 21405
R37K,S116F 6796 L133F,K118S 11666 D36H,R35A 16536 L84P,T96A 21406
R37K,S116G 6797 L133F,K118M 11667 D36H,R35E 16537 L84P,K100E 21407
R37K,S116N 6798 L133F,K118L 11668 D36H,R35F 16538 L84P,K101E 21408
R37K,S116K 6799 L133F,P121A 11669 D36H,R35G 16539 L84P,H102A 21409
R37K,S116H 6800 L133F,R122A 11670 D36H,R35I 16540 L84P,K105A 21410
R37K,S116Q 6801 L133F,R122E 11671 D36H,R35L 16541 L84P,K105E 21411
R37K,C117K 6802 L133F,H124A 11672 D36H,R35M 16542 L84P,K113A 21412
R37K,C117R 6803 L133F,Q127A 11673 D36H,R35P 16543 L84P,S116W 21413
R37K,C117S 6804 L133F,Q127K 11674 D36H,R35S 16544 L84P,S116Y 21414
R37K,C117Q 6805 L133F,K128E 11675 D36H,R35V 16545 L84P,S116F 21415 R37K,C117T 6806 L133G,K10E 11676 D36H,R37E 16546 L84P,S116G 21416
R37K,C117P 6807 L133G,Y15A 11677 D36H,R37A 16547 L84P,S116N 21417
R37K,C117A 6808 L133G,Y15F 11678 D36H,S38A 16548 L84P,S116K 21418
R37K,C117I 6809 L133G,S17A 11679 D36H,D39A 16549 L84P,S116H 21419
R37K,K118
6810 L133G,S17K 11680 D36H,Q40A 16550 L84P,S116Q 21420 W
R37K,K118H 6811 L133G,N18R 11681 D36H,Q43A 16551 L84P,C117K 21421
R37K,K118R 6812 L133G,H21A 11682 D36H,Q43E 16552 L84P,C117R 21422
R37K,K118S 6813 L133G,L26A 11683 D36H,Q43K 16553 L84P,C117S 21423
R37K,K118
6814 L133G,P27A 11684 D36H,S47A 16554 L84P,C117Q 21424 M
R37K,K118L 6815 L133G,D28K 11685 D36H,S47V 16555 L84P,C117T 21425
R37K,P121A 6816 L133G,T30A 11686 D36H,A48G 16556 L84P,C117P 21426
R37K,R122A 6817 L133G,T34A 11687 D36H,E53A 16557 L84P,C117A 21427
R37K,R122E 6818 L133G,R35A 11688 D36H,Y55A 16558 L84P,C117I 21428
R37K,H124A 6819 L133G,R35E 11689 D36H,Y55F 16559 L84P,K118W 21429
R37K,Q127A 6820 L133G,R35F 11690 D36H,K57A 16560 L84P,K118H 21430
R37K,Q127K 6821 L133G,R35G 11691 D36H,K57E 16561 L84P,K118R 21431
R37K,K128E 6822 L133G,R35I 11692 D36H,E60A 16562 L84P,K118S 21432
R37K,L133A 6823 L133G,R35L 11693 D36H,E60K 16563 L84P,K118M 21433
R37K,L133V 6824 L133G,R35M 11694 D36H,T61A 16564 L84P,K118L 21434
R37L,Q43D 6825 L133G,R35P 11695 D36H,Q63A 16565 L84P,P121A 21435
R37L,Q43F 6826 L133G,R35S 11696 D36H,Y64A 16566 L84P,R122A 21436
R37L,Q43G 6827 L133G,R35V 11697 D36H,D68A 16567 L84P,R122E 21437
R37L,Q43H 6828 L133G,D36A 11698 D36H,T78A 16568 L84P,H124A 21438
R37L,Q43I 6829 L133G,D36K 11699 D36H,P79A 16569 L84P,Q127A 21439
R37L,Q43L 6830 L133G,R37E 11700 D36H,E81K 16570 L84P,Q127K 21440
R37L,Q43M 6831 L133G,R37A 11701 D36H,L84A 16571 L84P,K128E 21441
R37L,Q43N 6832 L133G,S38A 11702 D36H,E87K 16572 L84P,L133A 21442
R37L,Q43P 6833 L133G,D39A 11703 D36H,E87Q 16573 L84P,L133V 21443
R37L,Q43S 6834 L133G,Q40A 11704 D36H,E87V 16574 L84Q,K10E 21444
R37L,Q43T 6835 L133G,Q43A 11705 D36H,E87D 16575 L84Q,Y15A 21445
R37L,Q43V 6836 L133G,Q43E 11706 D36H,R88L 16576 L84Q,Y15F 21446
R37L,Q43W 6837 L133G,Q43K 11707 D36H,R88Y 16577 L84Q,S17A 21447
R37L,Q43Y 6838 L133G,S47A 11708 D36H,R88D 16578 L84Q,S17K 21448
R37L,D28A 6839 L133G,S47V 11709 D36H,L89A 16579 L84Q,N18R 21449
R37L,E49A 6840 L133G,A48G 11710 D36H,E90K 16580 L84Q,H21A 21450
R37L,P121A 6841 L133G,E53A 11711 D36H,E91A 16581 L84Q,L26A 21451
R37L,K10E 6842 L133G,Y55A 11712 D36H,N92A 16582 L84Q,P27A 21452
R37L,Y15A 6843 L133G,Y55F 11713 D36H,Y94V 16583 L84Q,D28K 21453
R37L,Y15F 6844 L133G,K57A 11714 D36H,Y94A 16584 L84Q,T30A 21454
R37L,S17A 6845 L133G,K57E 11715 D36H,N95A 16585 L84Q,T34A 21455
R37L,S17K 6846 L133G,E60A 11716 D36H,N95V 16586 L84Q,R35A 21456
R37L,N18R 6847 L133G,E60K 11717 D36H,T96A 16587 L84Q,R35E 21457
R37L,H21A 6848 L133G,T61A 11718 D36H,K100E 16588 L84Q,R35F 21458
R37L,L26A 6849 L133G,Q63A 11719 D36H,K101E 16589 L84Q,R35G 21459
D36H,H102
R37L,P27A 6850 L133G,Y64A 11720 16590 L84Q,R35I 21460
A D36H,K105
R37L,D28K 6851 L133G,D68A 11721 16591 L84Q,R35L 21461
A
R37L,T30A 6852 L133G,T78A 11722 D36H,K105E 16592 L84Q,R35M 21462
D36H,K113
R37L,T34A 6853 L133G,P79A 11723 16593 L84Q,R35P 21463
A
D36H,S116
R37L,R35A 6854 L133G,E81K 11724 16594 L84Q,R35S 21464
W
R37L,R35E 6855 L133G,L84A 11725 D36H,S116Y 16595 L84Q,R35V 21465
R37L,R35F 6856 L133G,E87K 11726 D36H,S116F 16596 L84Q,D36A 21466
R37L,R35G 6857 L133G,E87Q 11727 D36H,S116G 16597 L84Q,D36K 21467
R37L,R35I 6858 L133G,E87V 11728 D36H,S116N 16598 L84Q,R37E 21468
R37L,R35L 6859 L133G,E87D 11729 D36H,S116K 16599 L84Q,R37A 21469
R37L,R35M 6860 L133G,R88L 11730 D36H,S116H 16600 L84Q,S38A 21470
R37L,R35P 6861 L133G,R88Y 11731 D36H,S116Q 16601 L84Q,D39A 21471
R37L,R35S 6862 L133G,R88D 11732 D36H,C117K 16602 L84Q,Q40A 21472
R37L,R35V 6863 L133G,L89A 11733 D36H,C117R 16603 L84Q,Q43A 21473
R37L,D36A 6864 L133G,E90K 11734 D36H,C117S 16604 L84Q,Q43E 21474
R37L,D36K 6865 L133G,E91A 11735 D36H,C117Q 16605 L84Q,Q43K 21475
R37L,S38A 6866 L133G,N92A 11736 D36H,C117T 16606 L84Q,S47A 21476
R37L,D39A 6867 L133G,Y94V 11737 D36H,C117P 16607 L84Q,S47V 21477
R37L,Q40A 6868 L133G,Y94A 11738 D36H,C117A 16608 L84Q,A48G 21478
R37L,Q43A 6869 L133G,N95A 11739 D36H,C117I 16609 L84Q,E53A 21479
D36H,K118
R37L,Q43E 6870 L133G,N95V 11740 16610 L84Q,Y55A 21480
W
D36H,K118
R37L,Q43K 6871 L133G,T96A 11741 16611 L84Q,Y55F 21481
H
R37L,S47A 6872 L133G,K100E 11742 D36H,K118R 16612 L84Q,K57A 21482
R37L,S47V 6873 L133G,K101E 11743 D36H,K118S 16613 L84Q,K57E 21483
D36H,K118
R37L,A48G 6874 L133G,H102A 11744 16614 L84Q,E60A 21484
M
R37L,E53A 6875 L133G,K105A 11745 D36H,K118L 16615 L84Q,E60K 21485
R37L,Y55A 6876 L133G,K105E 11746 D36H,P121A 16616 L84Q,T61A 21486
R37L,Y55F 6877 L133G,K113A 11747 D36H,R122A 16617 L84Q,Q63A 21487
L133G,S116
R37L,K57A 6878 11748 D36H,R122E 16618 L84Q,Y64A 21488
W
D36H,H124
R37L,K57E 6879 L133G,S116Y 11749 16619 L84Q,D68A 21489
A
D36H,Q127
R37L,E60A 6880 L133G,S116F 11750 16620 L84Q,T78A 21490
A
D36H,Q127
R37L,E60K 6881 L133G,S116G 11751 16621 L84Q,P79A 21491
K
R37L,T61A 6882 L133G,S116N 11752 D36H,K128E 16622 L84Q,E81K 21492
R37L,Q63A 6883 L133G,S116K 11753 D36H,L133A 16623 L84Q,E87K 21493
R37L,Y64A 6884 L133G,S116H 11754 D36H,L133V 16624 L84Q,E87Q 21494
R37L,D68A 6885 L133G,S116Q 11755 D36I,K10E 16625 L84Q,E87V 21495
R37L,T78A 6886 L133G,C117K 11756 D36I,Y15A 16626 L84Q,E87D 21496
R37L,P79A 6887 L133G,C117R 11757 D36I,Y15F 16627 L84Q,R88L 21497
R37L,E81K 6888 L133G,C117S 11758 D36I,S17A 16628 L84Q,R88Y 21498
R37L,L84A 6889 L133G,C117Q 11759 D36I,S17K 16629 L84Q,R88D 21499
R37L,E87K 6890 L133G,C117T 11760 D36I,N18R 16630 L84Q,L89A 21500
R37L,E87Q 6891 L133G,C117P 11761 D36I,H21A 16631 L84Q,E90K 21501 R37L,E87V 6892 L133G,C117A 11762 D36I,L26A 16632 L84Q,E91A 21502
R37L,E87D 6893 L133G,C117I 11763 D36I,P27A 16633 L84Q,N92A 21503
L133G,K118
R37L,R88L 6894 11764 D36I,D28K 16634 L84Q,Y94V 21504
W
R37L,R88Y 6895 L133G,K118H 11765 D36I,T30A 16635 L84Q,Y94A 21505
R37L,R88D 6896 L133G,K118R 11766 D36I,T34A 16636 L84Q,N95A 21506
R37L,L89A 6897 L133G,K118S 11767 D36I,R35A 16637 L84Q,N95V 21507
L133G,K118
R37L,E90K 6898 11768 D36I,R35E 16638 L84Q,T96A 21508
M
R37L,E91A 6899 L133G,K118L 11769 D36I,R35F 16639 L84Q,K100E 21509
R37L,N92A 6900 L133G,P121A 11770 D36I,R35G 16640 L84Q,K101E 21510
R37L,Y94V 6901 L133G,R122A 11771 D36I,R35I 16641 L84Q,H102A 21511
R37L,Y94A 6902 L133G,R122E 11772 D36I,R35L 16642 L84Q,K105A 21512
R37L,N95A 6903 L133G,H124A 11773 D36I,R35M 16643 L84Q,K105E 21513
R37L,N95V 6904 L133G,Q127A 11774 D36I,R35P 16644 L84Q,K113A 21514
R37L,T96A 6905 L133G,Q127K 11775 D36I,R35S 16645 L84Q,S116W 21515
R37L,K100E 6906 L133G,K128E 11776 D36I,R35V 16646 L84Q,S116Y 21516
R37L,K101E 6907 L133H,K10E 11777 D36I,R37E 16647 L84Q,S116F 21517
R37L,H102A 6908 L133H,Y15A 11778 D36I,R37A 16648 L84Q,S116G 21518
R37L,K105A 6909 L133H,Y15F 11779 D36I,S38A 16649 L84Q,S116N 21519
R37L,K105E 6910 L133H,S17A 11780 D36I,D39A 16650 L84Q,S116K 21520
R37L,K113A 6911 L133H,S17K 11781 D36I,Q40A 16651 L84Q,S116H 21521
R37L,S116W 6912 L133H,N18R 11782 D36I,Q43A 16652 L84Q,S116Q 21522
R37L,S116Y 6913 L133H,H21A 11783 D36I,Q43E 16653 L84Q,C117K 21523
R37L,S116F 6914 L133H,L26A 11784 D36I,Q43K 16654 L84Q,C117R 21524
R37L,S116G 6915 L133H,P27A 11785 D36I,S47A 16655 L84Q,C117S 21525
R37L,S116N 6916 L133H,D28K 11786 D36I,S47V 16656 L84Q,C117Q 21526
R37L,S116K 6917 L133H,T30A 11787 D36I,A48G 16657 L84Q,C117T 21527
R37L,S116H 6918 L133H,T34A 11788 D36I,E53A 16658 L84Q,C117P 21528
R37L,S116Q 6919 L133H,R35A 11789 D36I,Y55A 16659 L84Q,C117A 21529
R37L,C117K 6920 L133H,R35E 11790 D36I,Y55F 16660 L84Q,C117I 21530
R37L,C117R 6921 L133H,R35F 11791 D36I,K57A 16661 L84Q,K118W 21531
R37L,C117S 6922 L133H,R35G 11792 D36I,K57E 16662 L84Q,K118H 21532
R37L,C117Q 6923 L133H,R35I 11793 D36I,E60A 16663 L84Q,K118R 21533
R37L,C117T 6924 L133H,R35L 11794 D36I,E60K 16664 L84Q,K118S 21534
R37L,C117P 6925 L133H,R35M 11795 D36I,T61A 16665 L84Q,K118M 21535
R37L,C117A 6926 L133H,R35P 11796 D36I,Q63A 16666 L84Q,K118L 21536
R37L,C117I 6927 L133H,R35S 11797 D36I,Y64A 16667 L84Q,P121A 21537
R37L,K118W 6928 L133H,R35V 11798 D36I,D68A 16668 L84Q,R122A 21538
R37L,K118H 6929 L133H,D36A 11799 D36I,T78A 16669 L84Q,R122E 21539
R37L,K118R 6930 L133H,D36K 11800 D36I,P79A 16670 L84Q,H124A 21540
R37L,K118S 6931 L133H,R37E 11801 D36I,E81K 16671 L84Q,Q127A 21541
R37L,K118M 6932 L133H,R37A 11802 D36I,L84A 16672 L84Q,Q127K 21542
R37L,K118L 6933 L133H,S38A 11803 D36I,E87K 16673 L84Q,K128E 21543
R37L,P121A 6934 L133H,D39A 11804 D36I,E87Q 16674 L84Q,L133A 21544
R37L,R122A 6935 L133H,Q40A 11805 D36I,E87V 16675 L84Q,L133V 21545
R37L,R122E 6936 L133H,Q43A 11806 D36I,E87D 16676 L84R,K10E 21546 R37L,H124A 6937 L133H,Q43E 11807 D36I,R88L 16677 L84R,Y15A 21547
R37L,Q127A 6938 L133H,Q43K 11808 D36I,R88Y 16678 L84R,Y15F 21548
R37L,Q127K 6939 L133H,S47A 11809 D36I,R88D 16679 L84R,S17A 21549
R37L,K128E 6940 L133H,S47V 11810 D36I,L89A 16680 L84R,S17K 21550
R37L,L133A 6941 L133H,A48G 11811 D36I,E90K 16681 L84R,N18R 21551
R37L,L133V 6942 L133H,E53A 11812 D36I,E91A 16682 L84R,H21A 21552
R37M,Q43D 6943 L133H,Y55A 11813 D36I,N92A 16683 L84R,L26A 21553
R37M,Q43F 6944 L133H,Y55F 11814 D36I,Y94V 16684 L84R,P27A 21554
R37M,Q43G 6945 L133H,K57A 11815 D36I,Y94A 16685 L84R,D28K 21555
R37M,Q43H 6946 L133H,K57E 11816 D36I,N95A 16686 L84R,T30A 21556
R37M,Q43I 6947 L133H,E60A 11817 D36I,N95V 16687 L84R,T34A 21557
R37M,Q43L 6948 L133H,E60K 11818 D36I,T96A 16688 L84R,R35A 21558
R37M,Q43M 6949 L133H,T61A 11819 D36I,K100E 16689 L84R,R35E 21559
R37M,Q43N 6950 L133H,Q63A 11820 D36I,K101E 16690 L84R,R35F 21560
R37M,Q43P 6951 L133H,Y64A 11821 D36I,H102A 16691 L84R,R35G 21561
R37M,Q43S 6952 L133H,D68A 11822 D36I,K105A 16692 L84R,R35I 21562
R37M,Q43T 6953 L133H,T78A 11823 D36I,K105E 16693 L84R,R35L 21563
R37M,Q43V 6954 L133H,P79A 11824 D36I,K113A 16694 L84R,R35M 21564
R37M,Q43W 6955 L133H,E81K 11825 D36I,S116W 16695 L84R,R35P 21565
R37M,Q43Y 6956 L133H,L84A 11826 D36I,S116Y 16696 L84R,R35S 21566
R37M,D28A 6957 L133H,E87K 11827 D36I,S116F 16697 L84R,R35V 21567
R37M,E49A 6958 L133H,E87Q 11828 D36I,S116G 16698 L84R,D36A 21568
R37M,P121A 6959 L133H,E87V 11829 D36I,S116N 16699 L84R,D36K 21569
R37M,K10E 6960 L133H,E87D 11830 D36I,S116K 16700 L84R,R37E 21570
R37M,Y15A 6961 L133H,R88L 11831 D36I,S116H 16701 L84R,R37A 21571
R37M,Y15F 6962 L133H,R88Y 11832 D36I,S116Q 16702 L84R,S38A 21572
R37M,S17A 6963 L133H,R88D 11833 D36I,C117K 16703 L84R,D39A 21573
R37M,S17K 6964 L133H,L89A 11834 D36I,C117R 16704 L84R,Q40A 21574
R37M,N18R 6965 L133H,E90K 11835 D36I,C117S 16705 L84R,Q43A 21575
R37M,H21A 6966 L133H,E91A 11836 D36I,C117Q 16706 L84R,Q43E 21576
R37M,L26A 6967 L133H,N92A 11837 D36I,C117T 16707 L84R,Q43K 21577
R37M,P27A 6968 L133H,Y94V 11838 D36I,C117P 16708 L84R,S47A 21578
R37M,D28K 6969 L133H,Y94A 11839 D36I,C117A 16709 L84R,S47V 21579
R37M,T30A 6970 L133H,N95A 11840 D36I,C117I 16710 L84R,A48G 21580
R37M,T34A 6971 L133H,N95V 11841 D36I,K118W 16711 L84R,E53A 21581
R37M,R35A 6972 L133H,T96A 11842 D36I,K118H 16712 L84R,Y55A 21582
R37M,R35E 6973 L133H,K100E 11843 D36I,K118R 16713 L84R,Y55F 21583
R37M,R35F 6974 L133H,K101E 11844 D36I,K118S 16714 L84R,K57A 21584
R37M,R35G 6975 L133H,H102A 11845 D36I,K118M 16715 L84R,K57E 21585
R37M,R35I 6976 L133H,K105A 11846 D36I,K118L 16716 L84R,E60A 21586
R37M,R35L 6977 L133H,K105E 11847 D36I,P121A 16717 L84R,E60K 21587
R37M,R35M 6978 L133H,K113A 11848 D36I,R122A 16718 L84R,T61A 21588
L133H,S116
R37M,R35P 6979 11849 D36I,R122E 16719 L84R,Q63A 21589
W
R37M,R35S 6980 L133H,S116Y 11850 D36I,H124A 16720 L84R,Y64A 21590
R37M,R35V 6981 L133H,S116F 11851 D36I,Q127A 16721 L84R,D68A 21591
R37M,D36A 6982 L133H,S116G 11852 D36I,Q127K 16722 L84R,T78A 21592 R37M,D36K 6983 L133H,S116N 11853 D36I,K128E 16723 L84R,P79A 21593
R37M,S38A 6984 L133H,S116K 11854 D36I,L133A 16724 L84R,E81K 21594
R37M,D39A 6985 L133H,S116H 11855 D36I,L133V 16725 L84R,E87K 21595
R37M,Q40A 6986 L133H,S116Q 11856 D36L,K10E 16726 L84R,E87Q 21596
R37M,Q43A 6987 L133H,C117K 11857 D36L,Y15A 16727 L84R,E87V 21597
R37M,Q43E 6988 L133H,C117R 11858 D36L,Y15F 16728 L84R,E87D 21598
R37M,Q43K 6989 L133H,C117S 11859 D36L,S17A 16729 L84R,R88L 21599
R37M,S47A 6990 L133H,C117Q 11860 D36L,S17K 16730 L84R,R88Y 21600
R37M,S47V 6991 L133H,C117T 11861 D36L,N18R 16731 L84R,R88D 21601
R37M,A48G 6992 L133H,C117P 11862 D36L,H21A 16732 L84R,L89A 21602
R37M,E53A 6993 L133H,C117A 11863 D36L,L26A 16733 L84R,E90K 21603
R37M,Y55A 6994 L133H,C117I 11864 D36L,P27A 16734 L84R,E91A 21604
L133H,K118
R37M,Y55F 6995 11865 D36L,D28K 16735 L84R,N92A 21605
W
R37M,K57A 6996 L133H,K118H 11866 D36L,T30A 16736 L84R,Y94V 21606
R37M,K57E 6997 L133H,K118R 11867 D36L,T34A 16737 L84R,Y94A 21607
R37M,E60A 6998 L133H,K118S 11868 D36L,R35A 16738 L84R,N95A 21608
L133H,K118
R37M,E60K 6999 11869 D36L,R35E 16739 L84R,N95V 21609
M
R37M,T61A 7000 L133H,K118L 11870 D36L,R35F 16740 L84R,T96A 21610
R37M,Q63A 7001 L133H,P121A 11871 D36L,R35G 16741 L84R,K100E 21611
R37M,Y64A 7002 L133H,R122A 11872 D36L,R35I 16742 L84R,K101E 21612
R37M,D68A 7003 L133H,R122E 11873 D36L,R35L 16743 L84R,H102A 21613
R37M,T78A 7004 L133H,H124A 11874 D36L,R35M 16744 L84R,K105A 21614
R37M,P79A 7005 L133H,Q127A 11875 D36L,R35P 16745 L84R,K105E 21615
R37M,E81K 7006 L133H,Q127K 11876 D36L,R35S 16746 L84R,K113A 21616
R37M,L84A 7007 L133H,K128E 11877 D36L,R35V 16747 L84R,S116W 21617
R37M,E87K 7008 L133I,K10E 11878 D36L,R37E 16748 L84R,S116Y 21618
R37M,E87Q 7009 L133I,Y15A 11879 D36L,R37A 16749 L84R,S116F 21619
R37M,E87V 7010 L133I,Y15F 11880 D36L,S38A 16750 L84R,S116G 21620
R37M,E87D 7011 L133I,S17A 11881 D36L,D39A 16751 L84R,S116N 21621
R37M,R88L 7012 L133I,S17K 11882 D36L,Q40A 16752 L84R,S116K 21622
R37M,R88Y 7013 L133I,N18R 11883 D36L,Q43A 16753 L84R,S116H 21623
R37M,R88D 7014 L133I,H21A 11884 D36L,Q43E 16754 L84R,S116Q 21624
R37M,L89A 7015 L133I,L26A 11885 D36L,Q43K 16755 L84R,C117K 21625
R37M,E90K 7016 L133I,P27A 11886 D36L,S47A 16756 L84R,C117R 21626
R37M,E91A 7017 L133I,D28K 11887 D36L,S47V 16757 L84R,C117S 21627
R37M,N92A 7018 L133I,T30A 11888 D36L,A48G 16758 L84R,C117Q 21628
R37M,Y94V 7019 L133I,T34A 11889 D36L,E53A 16759 L84R,C117T 21629
R37M,Y94A 7020 L133I,R35A 11890 D36L,Y55A 16760 L84R,C117P 21630
R37M,N95A 7021 L133I,R35E 11891 D36L,Y55F 16761 L84R,C117A 21631
R37M,N95V 7022 L133I,R35F 11892 D36L,K57A 16762 L84R,C117I 21632
R37M,T96A 7023 L133I,R35G 11893 D36L,K57E 16763 L84R,K118W 21633
R37M,K100E 7024 L133I,R35I 11894 D36L,E60A 16764 L84R,K118H 21634
R37M,K101E 7025 L133I,R35L 11895 D36L,E60K 16765 L84R,K118R 21635
R37M,H102
7026 L133I,R35M 11896 D36L,T61A 16766 L84R,K118S 21636 A
R37M,K105 7027 L133I,R35P 11897 D36L,Q63A 16767 L84R,K118M 21637
Figure imgf000181_0001
R37N,Q43L 7066 L133I,E90K 11936 D36L,C117S 16806 L84S,Q40A 21676
R37N,Q43M 7067 L133I,E91A 11937 D36L,C117Q 16807 L84S,Q43A 21677
R37N,Q43N 7068 L133I,N92A 11938 D36L,C117T 16808 L84S,Q43E 21678
R37N,Q43P 7069 L133I,Y94V 11939 D36L,C117P 16809 L84S,Q43K 21679
R37N,Q43S 7070 L133I,Y94A 11940 D36L,C117A 16810 L84S,S47A 21680
R37N,Q43T 7071 L133I,N95A 11941 D36L,C117I 16811 L84S,S47V 21681
D36L,K118
R37N,Q43V 7072 L133I,N95V 11942 16812 L84S,A48G 21682
W
R37N,Q43W 7073 L133I,T96A 11943 D36L,K118H 16813 L84S,E53A 21683
R37N,Q43Y 7074 L133I,K100E 11944 D36L,K118R 16814 L84S,Y55A 21684
R37N,D28A 7075 L133I,K101E 11945 D36L,K118S 16815 L84S,Y55F 21685
D36L,K118
R37N,E49A 7076 L133I,H102A 11946 16816 L84S,K57A 21686
M
R37N,P121A 7077 L133I,K105A 11947 D36L,K118L 16817 L84S,K57E 21687
R37N,K10E 7078 L133I,K105E 11948 D36L,P121A 16818 L84S,E60A 21688
R37N,Y15A 7079 L133I,K113A 11949 D36L,R122A 16819 L84S,E60K 21689
R37N,Y15F 7080 L133I,S116W 11950 D36L,R122E 16820 L84S,T61A 21690
R37N,S17A 7081 L133I,S116Y 11951 D36L,H124A 16821 L84S,Q63A 21691
R37N,S17K 7082 L133I,S116F 11952 D36L,Q127A 16822 L84S,Y64A 21692
R37N,N18R 7083 L133I,S116G 11953 D36L,Q127K 16823 L84S,D68A 21693
R37N,H21A 7084 L133I,S116N 11954 D36L,K128E 16824 L84S,T78A 21694
R37N,L26A 7085 L133I,S116K 11955 D36L,L133A 16825 L84S,P79A 21695
R37N,P27A 7086 L133I,S116H 11956 D36L,L133V 16826 L84S,E81K 21696
R37N,D28K 7087 L133I,S116Q 11957 D36M,K10E 16827 L84S,E87K 21697
R37N,T30A 7088 L133I,C117K 11958 D36M,Y15A 16828 L84S,E87Q 21698
R37N,T34A 7089 L133I,C117R 11959 D36M,Y15F 16829 L84S,E87V 21699
R37N,R35A 7090 L133I,C117S 11960 D36M,S17A 16830 L84S,E87D 21700
R37N,R35E 7091 L133I,C117Q 11961 D36M,S17K 16831 L84S,R88L 21701
R37N,R35F 7092 L133I,C117T 11962 D36M,N18R 16832 L84S,R88Y 21702
R37N,R35G 7093 L133I,C117P 11963 D36M,H21A 16833 L84S,R88D 21703
R37N,R35I 7094 L133I,C117A 11964 D36M,L26A 16834 L84S,L89A 21704
R37N,R35L 7095 L133I,C117I 11965 D36M,P27A 16835 L84S,E90K 21705
R37N,R35M 7096 L133I,K118W 11966 D36M,D28K 16836 L84S,E91A 21706
R37N,R35P 7097 L133I,K118H 11967 D36M,T30A 16837 L84S,N92A 21707
R37N,R35S 7098 L133I,K118R 11968 D36M,T34A 16838 L84S,Y94V 21708
R37N,R35V 7099 L133I,K118S 11969 D36M,R35A 16839 L84S,Y94A 21709
R37N,D36A 7100 L133I,K118M 11970 D36M,R35E 16840 L84S,N95A 21710
R37N,D36K 7101 L133I,K118L 11971 D36M,R35F 16841 L84S,N95V 21711
R37N,S38A 7102 L133I,P121A 11972 D36M,R35G 16842 L84S,T96A 21712
R37N,D39A 7103 L133I,R122A 11973 D36M,R35I 16843 L84S,K100E 21713
R37N,Q40A 7104 L133I,R122E 11974 D36M,R35L 16844 L84S,K101E 21714
R37N,Q43A 7105 L133I,H124A 11975 D36M,R35M 16845 L84S,H102A 21715
R37N,Q43E 7106 L133I,Q127A 11976 D36M,R35P 16846 L84S,K105A 21716
R37N,Q43K 7107 L133I,Q127K 11977 D36M,R35S 16847 L84S,K105E 21717
R37N,S47A 7108 L133I,K128E 11978 D36M,R35V 16848 L84S,K113A 21718
R37N,S47V 7109 L133K,K10E 11979 D36M,R37E 16849 L84S,S116W 21719
R37N,A48G 7110 L133K,Y15A 11980 D36M,R37A 16850 L84S,S116Y 21720 R37N,E53A 7111 L133K,Y15F 11981 D36M,S38A 16851 L84S,S116F 21721
R37N,Y55A 7112 L133K,S17A 11982 D36M,D39A 16852 L84S,S116G 21722
R37N,Y55F 7113 L133K,S17K 11983 D36M,Q40A 16853 L84S,S116N 21723
R37N,K57A 7114 L133K,N18R 11984 D36M,Q43A 16854 L84S,S116K 21724
R37N,K57E 7115 L133K,H21A 11985 D36M,Q43E 16855 L84S,S116H 21725
R37N,E60A 7116 L133K,L26A 11986 D36M,Q43K 16856 L84S,S116Q 21726
R37N,E60K 7117 L133K,P27A 11987 D36M,S47A 16857 L84S,C117K 21727
R37N,T61A 7118 L133K,D28K 11988 D36M,S47V 16858 L84S,C117R 21728
R37N,Q63A 7119 L133K,T30A 11989 D36M,A48G 16859 L84S,C117S 21729
R37N,Y64A 7120 L133K,T34A 11990 D36M,E53A 16860 L84S,C117Q 21730
R37N,D68A 7121 L133K,R35A 11991 D36M,Y55A 16861 L84S,C117T 21731
R37N,T78A 7122 L133K,R35E 11992 D36M,Y55F 16862 L84S,C117P 21732
R37N,P79A 7123 L133K,R35F 11993 D36M,K57A 16863 L84S,C117A 21733
R37N,E81K 7124 L133K,R35G 11994 D36M,K57E 16864 L84S,C117I 21734
R37N,L84A 7125 L133K,R35I 11995 D36M,E60A 16865 L84S,K118W 21735
R37N,E87K 7126 L133K,R35L 11996 D36M,E60K 16866 L84S,K118H 21736
R37N,E87Q 7127 L133K,R35M 11997 D36M,T61A 16867 L84S,K118R 21737
R37N,E87V 7128 L133K,R35P 11998 D36M,Q63A 16868 L84S,K118S 21738
R37N,E87D 7129 L133K,R35S 11999 D36M,Y64A 16869 L84S,K118M 21739
R37N,R88L 7130 L133K,R35V 12000 D36M,D68A 16870 L84S,K118L 21740
R37N,R88Y 7131 L133K,D36A 12001 D36M,T78A 16871 L84S,P121A 21741
R37N,R88D 7132 L133K,D36K 12002 D36M,P79A 16872 L84S,R122A 21742
R37N,L89A 7133 L133K,R37E 12003 D36M,E81K 16873 L84S,R122E 21743
R37N,E90K 7134 L133K,R37A 12004 D36M,L84A 16874 L84S,H124A 21744
R37N,E91A 7135 L133K,S38A 12005 D36M,E87K 16875 L84S,Q127A 21745
R37N,N92A 7136 L133K,D39A 12006 D36M,E87Q 16876 L84S,Q127K 21746
R37N,Y94V 7137 L133K,Q40A 12007 D36M,E87V 16877 L84S,K128E 21747
R37N,Y94A 7138 L133K,Q43A 12008 D36M,E87D 16878 L84S,L133A 21748
R37N,N95A 7139 L133K,Q43E 12009 D36M,R88L 16879 L84S,L133V 21749
R37N,N95V 7140 L133K,Q43K 12010 D36M,R88Y 16880 L84T,K10E 21750
R37N,T96A 7141 L133K,S47A 12011 D36M,R88D 16881 L84T,Y15A 21751
R37N,K100E 7142 L133K,S47V 12012 D36M,L89A 16882 L84T,Y15F 21752
R37N,K101E 7143 L133K,A48G 12013 D36M,E90K 16883 L84T,S17A 21753
R37N,H102A 7144 L133K,E53A 12014 D36M,E91A 16884 L84T,S17K 21754
R37N,K105A 7145 L133K,Y55A 12015 D36M,N92A 16885 L84T,N18R 21755
R37N,K105E 7146 L133K,Y55F 12016 D36M,Y94V 16886 L84T,H21A 21756
R37N,K113A 7147 L133K,K57A 12017 D36M,Y94A 16887 L84T,L26A 21757
R37N,S116W 7148 L133K,K57E 12018 D36M,N95A 16888 L84T,P27A 21758
R37N,S116Y 7149 L133K,E60A 12019 D36M,N95V 16889 L84T,D28K 21759
R37N,S116F 7150 L133K,E60K 12020 D36M,T96A 16890 L84T,T30A 21760
D36M,K100
R37N,S116G 7151 L133K,T61A 12021 16891 L84T,T34A 21761
E
D36M,K101
R37N,S116N 7152 L133K,Q63A 12022 16892 L84T,R35A 21762
E
D36M,H102
R37N,S116K 7153 L133K,Y64A 12023 16893 L84T,R35E 21763
A
D36M,K105
R37N,S116H 7154 L133K,D68A 12024 16894 L84T,R35F 21764
A
Figure imgf000184_0001
D36M,L133
R37P,Q43N 7186 L133K,S116K 12056 16926 L84T,T78A 21796
A
D36M,L133
R37P,Q43P 7187 L133K,S116H 12057 16927 L84T,P79A 21797
V
R37P,Q43S 7188 L133K,S116Q 12058 D36N,K10E 16928 L84T,E81K 21798
R37P,Q43T 7189 L133K,C117K 12059 D36N,Y15A 16929 L84T,E87K 21799
R37P,Q43V 7190 L133K,C117R 12060 D36N,Y15F 16930 L84T,E87Q 21800
R37P,Q43W 7191 L133K,C117S 12061 D36N,S17A 16931 L84T,E87V 21801
R37P,Q43Y 7192 L133K,C117Q 12062 D36N,S17K 16932 L84T,E87D 21802
R37P,D28A 7193 L133K,C117T 12063 D36N,N18R 16933 L84T,R88L 21803
R37P,E49A 7194 L133K,C117P 12064 D36N,H21A 16934 L84T,R88Y 21804
R37P,P121A 7195 L133K,C117A 12065 D36N,L26A 16935 L84T,R88D 21805
R37P,K10E 7196 L133K,C117I 12066 D36N,P27A 16936 L84T,L89A 21806
L133K,K118
R37P,Y15A 7197 12067 D36N,D28K 16937 L84T,E90K 21807
W
R37P,Y15F 7198 L133K,K118H 12068 D36N,T30A 16938 L84T,E91A 21808
R37P,S17A 7199 L133K,K118R 12069 D36N,T34A 16939 L84T,N92A 21809
R37P,S17K 7200 L133K,K118S 12070 D36N,R35A 16940 L84T,Y94V 21810
L133K,K118
R37P,N18R 7201 12071 D36N,R35E 16941 L84T,Y94A 21811
M
R37P,H21A 7202 L133K,K118L 12072 D36N,R35F 16942 L84T,N95A 21812
R37P,L26A 7203 L133K,P121A 12073 D36N,R35G 16943 L84T,N95V 21813
R37P,P27A 7204 L133K,R122A 12074 D36N,R35I 16944 L84T,T96A 21814
R37P,D28K 7205 L133K,R122E 12075 D36N,R35L 16945 L84T,K100E 21815
R37P,T30A 7206 L133K,H124A 12076 D36N,R35M 16946 L84T,K101E 21816
R37P,T34A 7207 L133K,Q127A 12077 D36N,R35P 16947 L84T,H102A 21817
R37P,R35A 7208 L133K,Q127K 12078 D36N,R35S 16948 L84T,K105A 21818
R37P,R35E 7209 L133K,K128E 12079 D36N,R35V 16949 L84T,K105E 21819
R37P,R35F 7210 L133M,K10E 12080 D36N,R37E 16950 L84T,K113A 21820
R37P,R35G 7211 L133M,Y15A 12081 D36N,R37A 16951 L84T,S116W 21821
R37P,R35I 7212 L133M,Y15F 12082 D36N,S38A 16952 L84T,S116Y 21822
R37P,R35L 7213 L133M,S17A 12083 D36N,D39A 16953 L84T,S116F 21823
R37P,R35M 7214 L133M,S17K 12084 D36N,Q40A 16954 L84T,S116G 21824
R37P,R35P 7215 L133M,N18R 12085 D36N,Q43A 16955 L84T,S116N 21825
R37P,R35S 7216 L133M,H21A 12086 D36N,Q43E 16956 L84T,S116K 21826
R37P,R35V 7217 L133M,L26A 12087 D36N,Q43K 16957 L84T,S116H 21827
R37P,D36A 7218 L133M,P27A 12088 D36N,S47A 16958 L84T,S116Q 21828
R37P,D36K 7219 L133M,D28K 12089 D36N,S47V 16959 L84T,C117K 21829
R37P,S38A 7220 L133M,T30A 12090 D36N,A48G 16960 L84T,C117R 21830
R37P,D39A 7221 L133M,T34A 12091 D36N,E53A 16961 L84T,C117S 21831
R37P,Q40A 7222 L133M,R35A 12092 D36N,Y55A 16962 L84T,C117Q 21832
R37P,Q43A 7223 L133M,R35E 12093 D36N,Y55F 16963 L84T,C117T 21833
R37P,Q43E 7224 L133M,R35F 12094 D36N,K57A 16964 L84T,C117P 21834
R37P,Q43K 7225 L133M,R35G 12095 D36N,K57E 16965 L84T,C117A 21835
R37P,S47A 7226 L133M,R35I 12096 D36N,E60A 16966 L84T,C117I 21836
R37P,S47V 7227 L133M,R35L 12097 D36N,E60K 16967 L84T,K118W 21837
R37P,A48G 7228 L133M,R35M 12098 D36N,T61A 16968 L84T,K118H 21838
R37P,E53A 7229 L133M,R35P 12099 D36N,Q63A 16969 L84T,K118R 21839 R37P,Y55A 7230 L133M,R35S 12100 D36N,Y64A 16970 L84T,K118S 21840
R37P,Y55F 7231 L133M,R35V 12101 D36N,D68A 16971 L84T,K118M 21841
R37P,K57A 7232 L133M,D36A 12102 D36N,T78A 16972 L84T,K118L 21842
R37P,K57E 7233 L133M,D36K 12103 D36N,P79A 16973 L84T,P121A 21843
R37P,E60A 7234 L133M,R37E 12104 D36N,E81K 16974 L84T,R122A 21844
R37P,E60K 7235 L133M,R37A 12105 D36N,L84A 16975 L84T,R122E 21845
R37P,T61A 7236 L133M,S38A 12106 D36N,E87K 16976 L84T,H124A 21846
R37P,Q63A 7237 L133M,D39A 12107 D36N,E87Q 16977 L84T,Q127A 21847
R37P,Y64A 7238 L133M,Q40A 12108 D36N,E87V 16978 L84T,Q127K 21848
R37P,D68A 7239 L133M,Q43A 12109 D36N,E87D 16979 L84T,K128E 21849
R37P,T78A 7240 L133M,Q43E 12110 D36N,R88L 16980 L84T,L133A 21850
R37P,P79A 7241 L133M,Q43K 12111 D36N,R88Y 16981 L84T,L133V 21851
R37P,E81K 7242 L133M,S47A 12112 D36N,R88D 16982 L84V,K10E 21852
R37P,L84A 7243 L133M,S47V 12113 D36N,L89A 16983 L84V,Y15A 21853
R37P,E87K 7244 L133M,A48G 12114 D36N,E90K 16984 L84V,Y15F 21854
R37P,E87Q 7245 L133M,E53A 12115 D36N,E91A 16985 L84V,S17A 21855
R37P,E87V 7246 L133M,Y55A 12116 D36N,N92A 16986 L84V,S17K 21856
R37P,E87D 7247 L133M,Y55F 12117 D36N,Y94V 16987 L84V,N18R 21857
R37P,R88L 7248 L133M,K57A 12118 D36N,Y94A 16988 L84V,H21A 21858
R37P,R88Y 7249 L133M,K57E 12119 D36N,N95A 16989 L84V,L26A 21859
R37P,R88D 7250 L133M,E60A 12120 D36N,N95V 16990 L84V,P27A 21860
R37P,L89A 7251 L133M,E60K 12121 D36N,T96A 16991 L84V,D28K 21861
R37P,E90K 7252 L133M,T61A 12122 D36N,K100E 16992 L84V,T30A 21862
R37P,E91A 7253 L133M,Q63A 12123 D36N,K101E 16993 L84V,T34A 21863
D36N,H102
R37P,N92A 7254 L133M,Y64A 12124 16994 L84V,R35A 21864
A
D36N,K105
R37P,Y94V 7255 L133M,D68A 12125 16995 L84V,R35E 21865
A
R37P,Y94A 7256 L133M,T78A 12126 D36N,K105E 16996 L84V,R35F 21866
D36N,K113
R37P,N95A 7257 L133M,P79A 12127 16997 L84V,R35G 21867
A
D36N,S116
R37P,N95V 7258 L133M,E81K 12128 16998 L84V,R35I 21868
W
R37P,T96A 7259 L133M,L84A 12129 D36N,S116Y 16999 L84V,R35L 21869
R37P,K100E 7260 L133M,E87K 12130 D36N,S116F 17000 L84V,R35M 21870
R37P,K101E 7261 L133M,E87Q 12131 D36N,S116G 17001 L84V,R35P 21871
R37P,H102A 7262 L133M,E87V 12132 D36N,S116N 17002 L84V,R35S 21872
R37P,K105A 7263 L133M,E87D 12133 D36N,S116K 17003 L84V,R35V 21873
R37P,K105E 7264 L133M,R88L 12134 D36N,S116H 17004 L84V,D36A 21874
R37P,K113A 7265 L133M,R88Y 12135 D36N,S116Q 17005 L84V,D36K 21875
R37P,S116W 7266 L133M,R88D 12136 D36N,C117K 17006 L84V,R37E 21876
R37P,S116Y 7267 L133M,L89A 12137 D36N,C117R 17007 L84V,R37A 21877
R37P,S116F 7268 L133M,E90K 12138 D36N,C117S 17008 L84V,S38A 21878
R37P,S116G 7269 L133M,E91A 12139 D36N,C117Q 17009 L84V,D39A 21879
R37P,S116N 7270 L133M,N92A 12140 D36N,C117T 17010 L84V,Q40A 21880
R37P,S116K 7271 L133M,Y94V 12141 D36N,C117P 17011 L84V,Q43A 21881
R37P,S116H 7272 L133M,Y94A 12142 D36N,C117A 17012 L84V,Q43E 21882
R37P,S116Q 7273 L133M,N95A 12143 D36N,C117I 17013 L84V,Q43K 21883
Figure imgf000187_0001
A
L133M,Q127
R37Q,Q43V 7308 12178 D36P,R35P 17048 L84V,K101E 21918
A
L133M,Q127
R37Q,Q43W 7309 12179 D36P,R35S 17049 L84V,H102A 21919
K
L133M,K128
R37Q,Q43Y 7310 12180 D36P,R35V 17050 L84V,K105A 21920
E
R37Q,D28A 7311 L133N,K10E 12181 D36P,R37E 17051 L84V,K105E 21921
R37Q,E49A 7312 L133N,Y15A 12182 D36P,R37A 17052 L84V,K113A 21922
R37Q,P121A 7313 L133N,Y15F 12183 D36P,S38A 17053 L84V,S116W 21923
R37Q,K10E 7314 L133N,S17A 12184 D36P,D39A 17054 L84V,S116Y 21924
R37Q,Y15A 7315 L133N,S17K 12185 D36P,Q40A 17055 L84V,S116F 21925
R37Q,Y15F 7316 L133N,N18R 12186 D36P,Q43A 17056 L84V,S116G 21926
R37Q,S17A 7317 L133N,H21A 12187 D36P,Q43E 17057 L84V,S116N 21927
R37Q,S17K 7318 L133N,L26A 12188 D36P,Q43K 17058 L84V,S116K 21928
R37Q,N18R 7319 L133N,P27A 12189 D36P,S47A 17059 L84V,S116H 21929
R37Q,H21A 7320 L133N,D28K 12190 D36P,S47V 17060 L84V,S116Q 21930
R37Q,L26A 7321 L133N,T30A 12191 D36P,A48G 17061 L84V,C117K 21931
R37Q,P27A 7322 L133N,T34A 12192 D36P,E53A 17062 L84V,C117R 21932
R37Q,D28K 7323 L133N,R35A 12193 D36P,Y55A 17063 L84V,C117S 21933
R37Q,T30A 7324 L133N,R35E 12194 D36P,Y55F 17064 L84V,C117Q 21934
R37Q,T34A 7325 L133N,R35F 12195 D36P,K57A 17065 L84V,C117T 21935
R37Q,R35A 7326 L133N,R35G 12196 D36P,K57E 17066 L84V,C117P 21936
R37Q,R35E 7327 L133N,R35I 12197 D36P,E60A 17067 L84V,C117A 21937
R37Q,R35F 7328 L133N,R35L 12198 D36P,E60K 17068 L84V,C117I 21938
R37Q,R35G 7329 L133N,R35M 12199 D36P,T61A 17069 L84V,K118W 21939
R37Q,R35I 7330 L133N,R35P 12200 D36P,Q63A 17070 L84V,K118H 21940
R37Q,R35L 7331 L133N,R35S 12201 D36P,Y64A 17071 L84V,K118R 21941
R37Q,R35M 7332 L133N,R35V 12202 D36P,D68A 17072 L84V,K118S 21942
R37Q,R35P 7333 L133N,D36A 12203 D36P,T78A 17073 L84V,K118M 21943
R37Q,R35S 7334 L133N,D36K 12204 D36P,P79A 17074 L84V,K118L 21944
R37Q,R35V 7335 L133N,R37E 12205 D36P,E81K 17075 L84V,P121A 21945
R37Q,D36A 7336 L133N,R37A 12206 D36P,L84A 17076 L84V,R122A 21946
R37Q,D36K 7337 L133N,S38A 12207 D36P,E87K 17077 L84V,R122E 21947
R37Q,S38A 7338 L133N,D39A 12208 D36P,E87Q 17078 L84V,H124A 21948
R37Q,D39A 7339 L133N,Q40A 12209 D36P,E87V 17079 L84V,Q127A 21949
R37Q,Q40A 7340 L133N,Q43A 12210 D36P,E87D 17080 L84V,Q127K 21950
R37Q,Q43A 7341 L133N,Q43E 12211 D36P,R88L 17081 L84V,K128E 21951
R37Q,Q43E 7342 L133N,Q43K 12212 D36P,R88Y 17082 L84V,L133A 21952
R37Q,Q43K 7343 L133N,S47A 12213 D36P,R88D 17083 L84V,L133V 21953
R37Q,S47A 7344 L133N,S47V 12214 D36P,L89A 17084 L84W,K10E 21954
R37Q,S47V 7345 L133N,A48G 12215 D36P,E90K 17085 L84W,Y15A 21955
R37Q,A48G 7346 L133N,E53A 12216 D36P,E91A 17086 L84W,Y15F 21956
R37Q,E53A 7347 L133N,Y55A 12217 D36P,N92A 17087 L84W,S17A 21957
R37Q,Y55A 7348 L133N,Y55F 12218 D36P,Y94V 17088 L84W,S17K 21958
R37Q,Y55F 7349 L133N,K57A 12219 D36P,Y94A 17089 L84W,N18R 21959
R37Q,K57A 7350 L133N,K57E 12220 D36P,N95A 17090 L84W,H21A 21960
R37Q,K57E 7351 L133N,E60A 12221 D36P,N95V 17091 L84W,L26A 21961 R37Q,E60A 7352 L133N,E60K 12222 D36P,T96A 17092 L84W,P27A 21962
R37Q,E60K 7353 L133N,T61A 12223 D36P,K100E 17093 L84W,D28K 21963
R37Q,T61A 7354 L133N,Q63A 12224 D36P,K101E 17094 L84W,T30A 21964
R37Q,Q63A 7355 L133N,Y64A 12225 D36P,H102A 17095 L84W,T34A 21965
R37Q,Y64A 7356 L133N,D68A 12226 D36P,K105A 17096 L84W,R35A 21966
R37Q,D68A 7357 L133N,T78A 12227 D36P,K105E 17097 L84W,R35E 21967
R37Q,T78A 7358 L133N,P79A 12228 D36P,K113A 17098 L84W,R35F 21968
R37Q,P79A 7359 L133N,E81K 12229 D36P,S116W 17099 L84W,R35G 21969
R37Q,E81K 7360 L133N,L84A 12230 D36P,S116Y 17100 L84W,R35I 21970
R37Q,L84A 7361 L133N,E87K 12231 D36P,S116F 17101 L84W,R35L 21971
R37Q,E87K 7362 L133N,E87Q 12232 D36P,S116G 17102 L84W,R35M 21972
R37Q,E87Q 7363 L133N,E87V 12233 D36P,S116N 17103 L84W,R35P 21973
R37Q,E87V 7364 L133N,E87D 12234 D36P,S116K 17104 L84W,R35S 21974
R37Q,E87D 7365 L133N,R88L 12235 D36P,S116H 17105 L84W,R35V 21975
R37Q,R88L 7366 L133N,R88Y 12236 D36P,S116Q 17106 L84W,D36A 21976
R37Q,R88Y 7367 L133N,R88D 12237 D36P,C117K 17107 L84W,D36K 21977
R37Q,R88D 7368 L133N,L89A 12238 D36P,C117R 17108 L84W,R37E 21978
R37Q,L89A 7369 L133N,E90K 12239 D36P,C117S 17109 L84W,R37A 21979
R37Q,E90K 7370 L133N,E91A 12240 D36P,C117Q 17110 L84W,S38A 21980
R37Q,E91A 7371 L133N,N92A 12241 D36P,C117T 17111 L84W,D39A 21981
R37Q,N92A 7372 L133N,Y94V 12242 D36P,C117P 17112 L84W,Q40A 21982
R37Q,Y94V 7373 L133N,Y94A 12243 D36P,C117A 17113 L84W,Q43A 21983
R37Q,Y94A 7374 L133N,N95A 12244 D36P,C117I 17114 L84W,Q43E 21984
D36P,K118
R37Q,N95A 7375 L133N,N95V 12245 17115 L84W,Q43K 21985
W
R37Q,N95V 7376 L133N,T96A 12246 D36P,K118H 17116 L84W,S47A 21986
R37Q,T96A 7377 L133N,K100E 12247 D36P,K118R 17117 L84W,S47V 21987
R37Q,K100E 7378 L133N,K101E 12248 D36P,K118S 17118 L84W,A48G 21988
D36P,K118
R37Q,K101E 7379 L133N,H102A 12249 17119 L84W,E53A 21989
M
R37Q,H102A 7380 L133N,K105A 12250 D36P,K118L 17120 L84W,Y55A 21990
R37Q,K105A 7381 L133N,K105E 12251 D36P,P121A 17121 L84W,Y55F 21991
R37Q,K105E 7382 L133N,K113A 12252 D36P,R122A 17122 L84W,K57A 21992
L133N,S116
R37Q,K113A 7383 12253 D36P,R122E 17123 L84W,K57E 21993
W
R37Q,S116W 7384 L133N,S116Y 12254 D36P,H124A 17124 L84W,E60A 21994
R37Q,S116Y 7385 L133N,S116F 12255 D36P,Q127A 17125 L84W,E60K 21995
R37Q,S116F 7386 L133N,S116G 12256 D36P,Q127K 17126 L84W,T61A 21996
R37Q,S116G 7387 L133N,S116N 12257 D36P,K128E 17127 L84W,Q63A 21997
R37Q,S116N 7388 L133N,S116K 12258 D36P,L133A 17128 L84W,Y64A 21998
R37Q,S116K 7389 L133N,S116H 12259 D36P,L133V 17129 L84W,D68A 21999
R37Q,S116H 7390 L133N,S116Q 12260 D36Q,K10E 17130 L84W,T78A 22000
R37Q,S116Q 7391 L133N,C117K 12261 D36Q,Y15A 17131 L84W,P79A 22001
R37Q,C117K 7392 L133N,C117R 12262 D36Q,Y15F 17132 L84W,E81K 22002
R37Q,C117R 7393 L133N,C117S 12263 D36Q,S17A 17133 L84W,E87K 22003
R37Q,C117S 7394 L133N,C117Q 12264 D36Q,S17K 17134 L84W,E87Q 22004
R37Q,C117Q 7395 L133N,C117T 12265 D36Q,N18R 17135 L84W,E87V 22005
R37Q,C117T 7396 L133N,C117P 12266 D36Q,H21A 17136 L84W,E87D 22006 R37Q,C117P 7397 L133N,C117A 12267 D36Q,L26A 17137 L84W,R88L 22007
R37Q,C117A 7398 L133N,C117I 12268 D36Q,P27A 17138 L84W,R88Y 22008
L133N,K118
R37Q,C117I 7399 12269 D36Q,D28K 17139 L84W,R88D 22009
W
R37Q,K118
7400 L133N,K118H 12270 D36Q,T30A 17140 L84W,L89A 22010 W
R37Q,K118H 7401 L133N,K118R 12271 D36Q,T34A 17141 L84W,E90K 22011
R37Q,K118R 7402 L133N,K118S 12272 D36Q,R35A 17142 L84W,E91A 22012
L133N,K118
R37Q,K118S 7403 12273 D36Q,R35E 17143 L84W,N92A 22013
M
R37Q,K118
7404 L133N,K118L 12274 D36Q,R35F 17144 L84W,Y94V 22014 M
R37Q,K118L 7405 L133N,P121A 12275 D36Q,R35G 17145 L84W,Y94A 22015
R37Q,P121A 7406 L133N,R122A 12276 D36Q,R35I 17146 L84W,N95A 22016
R37Q,R122A 7407 L133N,R122E 12277 D36Q,R35L 17147 L84W,N95V 22017
R37Q,R122E 7408 L133N,H124A 12278 D36Q,R35M 17148 L84W,T96A 22018
R37Q,H124A 7409 L133N,Q127A 12279 D36Q,R35P 17149 L84W,K100E 22019
R37Q,Q127A 7410 L133N,Q127K 12280 D36Q,R35S 17150 L84W,K101E 22020
R37Q,Q127K 7411 L133N,K128E 12281 D36Q,R35V 17151 L84W,H102A 22021
R37Q,K128E 7412 L133P,K10E 12282 D36Q,R37E 17152 L84W,K105A 22022
R37Q,L133A 7413 L133P,Y15A 12283 D36Q,R37A 17153 L84W,K105E 22023
R37Q,L133V 7414 L133P,Y15F 12284 D36Q,S38A 17154 L84W,K113A 22024
R37S,Q43D 7415 L133P,S17A 12285 D36Q,D39A 17155 L84W,S116W 22025
R37S,Q43F 7416 L133P,S17K 12286 D36Q,Q40A 17156 L84W,S116Y 22026
R37S,Q43G 7417 L133P,N18R 12287 D36Q,Q43A 17157 L84W,S116F 22027
R37S,Q43H 7418 L133P,H21A 12288 D36Q,Q43E 17158 L84W,S116G 22028
R37S,Q43I 7419 L133P,L26A 12289 D36Q,Q43K 17159 L84W,S116N 22029
R37S,Q43L 7420 L133P,P27A 12290 D36Q,S47A 17160 L84W,S116K 22030
R37S,Q43M 7421 L133P,D28K 12291 D36Q,S47V 17161 L84W,S116H 22031
R37S,Q43N 7422 L133P,T30A 12292 D36Q,A48G 17162 L84W,S116Q 22032
R37S,Q43P 7423 L133P,T34A 12293 D36Q,E53A 17163 L84W,C117K 22033
R37S,Q43S 7424 L133P,R35A 12294 D36Q,Y55A 17164 L84W,C117R 22034
R37S,Q43T 7425 L133P,R35E 12295 D36Q,Y55F 17165 L84W,C117S 22035
R37S,Q43V 7426 L133P,R35F 12296 D36Q,K57A 17166 L84W,C117Q 22036
R37S,Q43W 7427 L133P,R35G 12297 D36Q,K57E 17167 L84W,C117T 22037
R37S,Q43Y 7428 L133P,R35I 12298 D36Q,E60A 17168 L84W,C117P 22038
R37S,D28A 7429 L133P,R35L 12299 D36Q,E60K 17169 L84W,C117A 22039
R37S,E49A 7430 L133P,R35M 12300 D36Q,T61A 17170 L84W,C117I 22040
R37S,P121A 7431 L133P,R35P 12301 D36Q,Q63A 17171 L84W,K118W 22041
R37S,K10E 7432 L133P,R35S 12302 D36Q,Y64A 17172 L84W,K118H 22042
R37S,Y15A 7433 L133P,R35V 12303 D36Q,D68A 17173 L84W,K118R 22043
R37S,Y15F 7434 L133P,D36A 12304 D36Q,T78A 17174 L84W,K118S 22044
R37S,S17A 7435 L133P,D36K 12305 D36Q,P79A 17175 L84W,K118M 22045
R37S,S17K 7436 L133P,R37E 12306 D36Q,E81K 17176 L84W,K118L 22046
R37S,N18R 7437 L133P,R37A 12307 D36Q,L84A 17177 L84W,P121A 22047
R37S,H21A 7438 L133P,S38A 12308 D36Q,E87K 17178 L84W,R122A 22048
R37S,L26A 7439 L133P,D39A 12309 D36Q,E87Q 17179 L84W,R122E 22049
R37S,P27A 7440 L133P,Q40A 12310 D36Q,E87V 17180 L84W,H124A 22050 R37S,D28K 7441 L133P,Q43A 12311 D36Q,E87D 17181 L84W,Q127A 22051
R37S,T30A 7442 L133P,Q43E 12312 D36Q,R88L 17182 L84W,Q127K 22052
R37S,T34A 7443 L133P,Q43K 12313 D36Q,R88Y 17183 L84W,K128E 22053
R37S,R35A 7444 L133P,S47A 12314 D36Q,R88D 17184 L84W,L133A 22054
R37S,R35E 7445 L133P,S47V 12315 D36Q,L89A 17185 L84W,L133V 22055
R37S,R35F 7446 L133P,A48G 12316 D36Q,E90K 17186 L84Y,K10E 22056
R37S,R35G 7447 L133P,E53A 12317 D36Q,E91A 17187 L84Y,Y15A 22057
R37S,R35I 7448 L133P,Y55A 12318 D36Q,N92A 17188 L84Y,Y15F 22058
R37S,R35L 7449 L133P,Y55F 12319 D36Q,Y94V 17189 L84Y,S17A 22059
R37S,R35M 7450 L133P,K57A 12320 D36Q,Y94A 17190 L84Y,S17K 22060
R37S,R35P 7451 L133P,K57E 12321 D36Q,N95A 17191 L84Y,N18R 22061
R37S,R35S 7452 L133P,E60A 12322 D36Q,N95V 17192 L84Y,H21A 22062
R37S,R35V 7453 L133P,E60K 12323 D36Q,T96A 17193 L84Y,L26A 22063
R37S,D36A 7454 L133P,T61A 12324 D36Q,K100E 17194 L84Y,P27A 22064
R37S,D36K 7455 L133P,Q63A 12325 D36Q,K101E 17195 L84Y,D28K 22065
D36Q,H102
R37S,S38A 7456 L133P,Y64A 12326 17196 L84Y,T30A 22066
A
D36Q,K105
R37S,D39A 7457 L133P,D68A 12327 17197 L84Y,T34A 22067
A
R37S,Q40A 7458 L133P,T78A 12328 D36Q,K105E 17198 L84Y,R35A 22068
D36Q,K113
R37S,Q43A 7459 L133P,P79A 12329 17199 L84Y,R35E 22069
A
D36Q,S116
R37S,Q43E 7460 L133P,E81K 12330 17200 L84Y,R35F 22070
W
R37S,Q43K 7461 L133P,L84A 12331 D36Q,S116Y 17201 L84Y,R35G 22071
R37S,S47A 7462 L133P,E87K 12332 D36Q,S116F 17202 L84Y,R35I 22072
R37S,S47V 7463 L133P,E87Q 12333 D36Q,S116G 17203 L84Y,R35L 22073
R37S,A48G 7464 L133P,E87V 12334 D36Q,S116N 17204 L84Y,R35M 22074
R37S,E53A 7465 L133P,E87D 12335 D36Q,S116K 17205 L84Y,R35P 22075
R37S,Y55A 7466 L133P,R88L 12336 D36Q,S116H 17206 L84Y,R35S 22076
R37S,Y55F 7467 L133P,R88Y 12337 D36Q,S116Q 17207 L84Y,R35V 22077
R37S,K57A 7468 L133P,R88D 12338 D36Q,C117K 17208 L84Y,D36A 22078
R37S,K57E 7469 L133P,L89A 12339 D36Q,C117R 17209 L84Y,D36K 22079
R37S,E60A 7470 L133P,E90K 12340 D36Q,C117S 17210 L84Y,R37E 22080
R37S,E60K 7471 L133P,E91A 12341 D36Q,C117Q 17211 L84Y,R37A 22081
R37S,T61A 7472 L133P,N92A 12342 D36Q,C117T 17212 L84Y,S38A 22082
R37S,Q63A 7473 L133P,Y94V 12343 D36Q,C117P 17213 L84Y,D39A 22083
R37S,Y64A 7474 L133P,Y94A 12344 D36Q,C117A 17214 L84Y,Q40A 22084
R37S,D68A 7475 L133P,N95A 12345 D36Q,C117I 17215 L84Y,Q43A 22085
D36Q,K118
R37S,T78A 7476 L133P,N95V 12346 17216 L84Y,Q43E 22086
W
D36Q,K118
R37S,P79A 7477 L133P,T96A 12347 17217 L84Y,Q43K 22087
H
R37S,E81K 7478 L133P,K100E 12348 D36Q,K118R 17218 L84Y,S47A 22088
R37S,L84A 7479 L133P,K101E 12349 D36Q,K118S 17219 L84Y,S47V 22089
D36Q,K118
R37S,E87K 7480 L133P,H102A 12350 17220 L84Y,A48G 22090
M
R37S,E87Q 7481 L133P,K105A 12351 D36Q,K118L 17221 L84Y,E53A 22091
R37S,E87V 7482 L133P,K105E 12352 D36Q,P121A 17222 L84Y,Y55A 22092 R37S,E87D 7483 L133P,K113A 12353 D36Q,R122A 17223 L84Y,Y55F 22093
R37S,R88L 7484 L133P,S116W 12354 D36Q,R122E 17224 L84Y,K57A 22094
D36Q,H124
R37S,R88Y 7485 L133P,S116Y 12355 17225 L84Y,K57E 22095
A
D36Q,Q127
R37S,R88D 7486 L133P,S116F 12356 17226 L84Y,E60A 22096
A
D36Q,Q127
R37S,L89A 7487 L133P,S116G 12357 17227 L84Y,E60K 22097
K
R37S,E90K 7488 L133P,S116N 12358 D36Q,K128E 17228 L84Y,T61A 22098
R37S,E91A 7489 L133P,S116K 12359 D36Q,L133A 17229 L84Y,Q63A 22099
R37S,N92A 7490 L133P,S116H 12360 D36Q,L133V 17230 L84Y,Y64A 22100
R37S,Y94V 7491 L133P,S116Q 12361 D36R,K10E 17231 L84Y,D68A 22101
R37S,Y94A 7492 L133P,C117K 12362 D36R,Y15A 17232 L84Y,T78A 22102
R37S,N95A 7493 L133P,C117R 12363 D36R,Y15F 17233 L84Y,P79A 22103
R37S,N95V 7494 L133P,C117S 12364 D36R,S17A 17234 L84Y,E81K 22104
R37S,T96A 7495 L133P,C117Q 12365 D36R,S17K 17235 L84Y,E87K 22105
R37S,K100E 7496 L133P,C117T 12366 D36R,N18R 17236 L84Y,E87Q 22106
R37S,K101E 7497 L133P,C117P 12367 D36R,H21A 17237 L84Y,E87V 22107
R37S,H102A 7498 L133P,C117A 12368 D36R,L26A 17238 L84Y,E87D 22108
R37S,K105A 7499 L133P,C117I 12369 D36R,P27A 17239 L84Y,R88L 22109
L133P,K118
R37S,K105E 7500 12370 D36R,D28K 17240 L84Y,R88Y 22110
W
R37S,K113A 7501 L133P,K118H 12371 D36R,T30A 17241 L84Y,R88D 22111
R37S,S116W 7502 L133P,K118R 12372 D36R,T34A 17242 L84Y,L89A 22112
R37S,S116Y 7503 L133P,K118S 12373 D36R,R35A 17243 L84Y,E90K 22113
R37S,S116F 7504 L133P,K118M 12374 D36R,R35E 17244 L84Y,E91A 22114
R37S,S116G 7505 L133P,K118L 12375 D36R,R35F 17245 L84Y,N92A 22115
R37S,S116N 7506 L133P,P121A 12376 D36R,R35G 17246 L84Y,Y94V 22116
R37S,S116K 7507 L133P,R122A 12377 D36R,R35I 17247 L84Y,Y94A 22117
R37S,S116H 7508 L133P,R122E 12378 D36R,R35L 17248 L84Y,N95A 22118
R37S,S116Q 7509 L133P,H124A 12379 D36R,R35M 17249 L84Y,N95V 22119
R37S,C117K 7510 L133P,Q127A 12380 D36R,R35P 17250 L84Y,T96A 22120
R37S,C117R 7511 L133P,Q127K 12381 D36R,R35S 17251 L84Y,K100E 22121
R37S,C117S 7512 L133P,K128E 12382 D36R,R35V 17252 L84Y,K101E 22122
R37S,C117Q 7513 L133Q,K10E 12383 D36R,R37E 17253 L84Y,H102A 22123
R37S,C117T 7514 L133Q,Y15A 12384 D36R,R37A 17254 L84Y,K105A 22124
R37S,C117P 7515 L133Q,Y15F 12385 D36R,S38A 17255 L84Y,K105E 22125
R37S,C117A 7516 L133Q,S17A 12386 D36R,D39A 17256 L84Y,K113A 22126
R37S,C117I 7517 L133Q,S17K 12387 D36R,Q40A 17257 L84Y,S116W 22127
R37S,K118W 7518 L133Q,N18R 12388 D36R,Q43A 17258 L84Y,S116Y 22128
R37S,K118H 7519 L133Q,H21A 12389 D36R,Q43E 17259 L84Y,S116F 22129
R37S,K118R 7520 L133Q,L26A 12390 D36R,Q43K 17260 L84Y,S116G 22130
R37S,K118S 7521 L133Q,P27A 12391 D36R,S47A 17261 L84Y,S116N 22131
R37S,K118M 7522 L133Q,D28K 12392 D36R,S47V 17262 L84Y,S116K 22132
R37S,K118L 7523 L133Q,T30A 12393 D36R,A48G 17263 L84Y,S116H 22133
R37S,P121A 7524 L133Q,T34A 12394 D36R,E53A 17264 L84Y,S116Q 22134
R37S,R122A 7525 L133Q,R35A 12395 D36R,Y55A 17265 L84Y,C117K 22135
R37S,R122E 7526 L133Q,R35E 12396 D36R,Y55F 17266 L84Y,C117R 22136 R37S,H124A 7527 L133Q,R35F 12397 D36R,K57A 17267 L84Y,C117S 22137
R37S,Q127A 7528 L133Q,R35G 12398 D36R,K57E 17268 L84Y,C117Q 22138
R37S,Q127K 7529 L133Q,R35I 12399 D36R,E60A 17269 L84Y,C117T 22139
R37S,K128E 7530 L133Q,R35L 12400 D36R,E60K 17270 L84Y,C117P 22140
R37S,L133A 7531 L133Q,R35M 12401 D36R,T61A 17271 L84Y,C117A 22141
R37S,L133V 7532 L133Q,R35P 12402 D36R,Q63A 17272 L84Y,C117I 22142
R37T,Q43D 7533 L133Q,R35S 12403 D36R,Y64A 17273 L84Y,K118W 22143
R37T,Q43F 7534 L133Q,R35V 12404 D36R,D68A 17274 L84Y,K118H 22144
R37T,Q43G 7535 L133Q,D36A 12405 D36R,T78A 17275 L84Y,K118R 22145
R37T,Q43H 7536 L133Q,D36K 12406 D36R,P79A 17276 L84Y,K118S 22146
R37T,Q43I 7537 L133Q,R37E 12407 D36R,E81K 17277 L84Y,K118M 22147
R37T,Q43L 7538 L133Q,R37A 12408 D36R,L84A 17278 L84Y,K118L 22148
R37T,Q43M 7539 L133Q,S38A 12409 D36R,E87K 17279 L84Y,P121A 22149
R37T,Q43N 7540 L133Q,D39A 12410 D36R,E87Q 17280 L84Y,R122A 22150
R37T,Q43P 7541 L133Q,Q40A 12411 D36R,E87V 17281 L84Y,R122E 22151
R37T,Q43S 7542 L133Q,Q43A 12412 D36R,E87D 17282 L84Y,H124A 22152
R37T,Q43T 7543 L133Q,Q43E 12413 D36R,R88L 17283 L84Y,Q127A 22153
R37T,Q43V 7544 L133Q,Q43K 12414 D36R,R88Y 17284 L84Y,Q127K 22154
R37T,Q43W 7545 L133Q,S47A 12415 D36R,R88D 17285 L84Y,K128E 22155
R37T,Q43Y 7546 L133Q,S47V 12416 D36R,L89A 17286 L84Y,L133A 22156
R37T,D28A 7547 L133Q,A48G 12417 D36R,E90K 17287 L84Y,L133V 22157
R37T,E49A 7548 L133Q,E53A 12418 D36R,E91A 17288 K101D,K10E 22158
R37T,P121A 7549 L133Q,Y55A 12419 D36R,N92A 17289 K101D,Y15A 22159
R37T,K10E 7550 L133Q,Y55F 12420 D36R,Y94V 17290 K101D,Y15F 22160
R37T,Y15A 7551 L133Q,K57A 12421 D36R,Y94A 17291 K101D,S17A 22161
R37T,Y15F 7552 L133Q,K57E 12422 D36R,N95A 17292 K101D,S17K 22162
R37T,S17A 7553 L133Q,E60A 12423 D36R,N95V 17293 K101D,N18R 22163
R37T,S17K 7554 L133Q,E60K 12424 D36R,T96A 17294 K101D,H21A 22164
R37T,N18R 7555 L133Q,T61A 12425 D36R,K100E 17295 K101D,L26A 22165
R37T,H21A 7556 L133Q,Q63A 12426 D36R,K101E 17296 K101D,P27A 22166
R37T,L26A 7557 L133Q,Y64A 12427 D36R,H102A 17297 K101D,D28K 22167
R37T,P27A 7558 L133Q,D68A 12428 D36R,K105A 17298 K101D,T30A 22168
R37T,D28K 7559 L133Q,T78A 12429 D36R,K105E 17299 K101D,T34A 22169
R37T,T30A 7560 L133Q,P79A 12430 D36R,K113A 17300 K101D,R35A 22170
D36R,S116
R37T,T34A 7561 L133Q,E81K 12431 17301 K101D,R35E 22171
W
R37T,R35A 7562 L133Q,L84A 12432 D36R,S116Y 17302 K101D,R35F 22172
R37T,R35E 7563 L133Q,E87K 12433 D36R,S116F 17303 K101D,R35G 22173
R37T,R35F 7564 L133Q,E87Q 12434 D36R,S116G 17304 K101D,R35I 22174
R37T,R35G 7565 L133Q,E87V 12435 D36R,S116N 17305 K101D,R35L 22175
R37T,R35I 7566 L133Q,E87D 12436 D36R,S116K 17306 K101D,R35M 22176
R37T,R35L 7567 L133Q,R88L 12437 D36R,S116H 17307 K101D,R35P 22177
R37T,R35M 7568 L133Q,R88Y 12438 D36R,S116Q 17308 K101D,R35S 22178
R37T,R35P 7569 L133Q,R88D 12439 D36R,C117K 17309 K101D,R35V 22179
R37T,R35S 7570 L133Q,L89A 12440 D36R,C117R 17310 K101D,D36A 22180
R37T,R35V 7571 L133Q,E90K 12441 D36R,C117S 17311 K101D,D36K 22181
R37T,D36A 7572 L133Q,E91A 12442 D36R,C117Q 17312 K101D,R37E 22182 R37T,D36K 7573 L133Q,N92A 12443 D36R,C117T 17313 K101D,R37A 22183
R37T,S38A 7574 L133Q,Y94V 12444 D36R,C117P 17314 K101D,S38A 22184
R37T,D39A 7575 L133Q,Y94A 12445 D36R,C117A 17315 K101D,D39A 22185
R37T,Q40A 7576 L133Q,N95A 12446 D36R,C117I 17316 K101D,Q40A 22186
D36R,K118
R37T,Q43A 7577 L133Q,N95V 12447 17317 K101D,Q43A 22187
W
R37T,Q43E 7578 L133Q,T96A 12448 D36R,K118H 17318 K101D,Q43E 22188
R37T,Q43K 7579 L133Q,K100E 12449 D36R,K118R 17319 K101D,Q43K 22189
R37T,S47A 7580 L133Q,K101E 12450 D36R,K118S 17320 K101D,S47A 22190
D36R,K118
R37T,S47V 7581 L133Q,H102A 12451 17321 K101D,S47V 22191
M
R37T,A48G 7582 L133Q,K105A 12452 D36R,K118L 17322 K101D,A48G 22192
R37T,E53A 7583 L133Q,K105E 12453 D36R,P121A 17323 K101D,E53A 22193
R37T,Y55A 7584 L133Q,K113A 12454 D36R,R122A 17324 K101D,Y55A 22194
L133Q,S116
R37T,Y55F 7585 12455 D36R,R122E 17325 K101D,Y55F 22195
W
R37T,K57A 7586 L133Q,S116Y 12456 D36R,H124A 17326 K101D,K57A 22196
R37T,K57E 7587 L133Q,S116F 12457 D36R,Q127A 17327 K101D,K57E 22197
R37T,E60A 7588 L133Q,S116G 12458 D36R,Q127K 17328 K101D,E60A 22198
R37T,E60K 7589 L133Q,S116N 12459 D36R,K128E 17329 K101D,E60K 22199
R37T,T61A 7590 L133Q,S116K 12460 D36R,L133A 17330 K101D,T61A 22200
R37T,Q63A 7591 L133Q,S116H 12461 D36R,L133V 17331 K101D,Q63A 22201
R37T,Y64A 7592 L133Q,S116Q 12462 D36S,K10E 17332 K101D,Y64A 22202
R37T,D68A 7593 L133Q,C117K 12463 D36S,Y15A 17333 K101D,D68A 22203
R37T,T78A 7594 L133Q,C117R 12464 D36S,Y15F 17334 K101D,T78A 22204
R37T,P79A 7595 L133Q,C117S 12465 D36S,S17A 17335 K101D,P79A 22205
R37T,E81K 7596 L133Q,C117Q 12466 D36S,S17K 17336 K101D,E81K 22206
R37T,L84A 7597 L133Q,C117T 12467 D36S,N18R 17337 K101D,L84A 22207
R37T,E87K 7598 L133Q,C117P 12468 D36S,H21A 17338 K101D,E87K 22208
R37T,E87Q 7599 L133Q,C117A 12469 D36S,L26A 17339 K101D,E87Q 22209
R37T,E87V 7600 L133Q,C117I 12470 D36S,P27A 17340 K101D,E87V 22210
L133Q,K118
R37T,E87D 7601 12471 D36S,D28K 17341 K101D,E87D 22211
W
R37T,R88L 7602 L133Q,K118H 12472 D36S,T30A 17342 K101D,R88L 22212
R37T,R88Y 7603 L133Q,K118R 12473 D36S,T34A 17343 K101D,R88Y 22213
R37T,R88D 7604 L133Q,K118S 12474 D36S,R35A 17344 K101D,R88D 22214
L133Q,K118
R37T,L89A 7605 12475 D36S,R35E 17345 K101D,L89A 22215
M
R37T,E90K 7606 L133Q,K118L 12476 D36S,R35F 17346 K101D,E90K 22216
R37T,E91A 7607 L133Q,P121A 12477 D36S,R35G 17347 K101D,E91A 22217
R37T,N92A 7608 L133Q,R122A 12478 D36S,R35I 17348 K101D,N92A 22218
R37T,Y94V 7609 L133Q,R122E 12479 D36S,R35L 17349 K101D,Y94V 22219
R37T,Y94A 7610 L133Q,H124A 12480 D36S,R35M 17350 K101D,Y94A 22220
R37T,N95A 7611 L133Q,Q127A 12481 D36S,R35P 17351 K101D,N95A 22221
R37T,N95V 7612 L133Q,Q127K 12482 D36S,R35S 17352 K101D,N95V 22222
R37T,T96A 7613 L133Q,K128E 12483 D36S,R35V 17353 K101D,T96A 22223
R37T,K100E 7614 L133R,K10E 12484 D36S,R37E 17354 K101D,K100E 22224
K101D,H102
R37T,K101E 7615 L133R,Y15A 12485 D36S,R37A 17355 22225
A
Figure imgf000195_0001
R37V,Q43H 7654 L133R,E60A 12524 D36S,N95V 17394 K101F,S17K 22264
R37V,Q43I 7655 L133R,E60K 12525 D36S,T96A 17395 K101F,N18R 22265
R37V,Q43L 7656 L133R,T61A 12526 D36S,K100E 17396 K101F,H21A 22266
R37V,Q43M 7657 L133R,Q63A 12527 D36S,K101E 17397 K101F,L26A 22267
R37V,Q43N 7658 L133R,Y64A 12528 D36S,H102A 17398 K101F,P27A 22268
R37V,Q43P 7659 L133R,D68A 12529 D36S,K105A 17399 K101F,D28K 22269
R37V,Q43S 7660 L133R,T78A 12530 D36S,K105E 17400 K101F,T30A 22270
R37V,Q43T 7661 L133R,P79A 12531 D36S,K113A 17401 K101F,T34A 22271
R37V,Q43V 7662 L133R,E81K 12532 D36S,S116W 17402 K101F,R35A 22272
R37V,Q43W 7663 L133R,L84A 12533 D36S,S116Y 17403 K101F,R35E 22273
R37V,Q43Y 7664 L133R,E87K 12534 D36S,S116F 17404 K101F,R35F 22274
R37V,D28A 7665 L133R,E87Q 12535 D36S,S116G 17405 K101F,R35G 22275
R37V,E49A 7666 L133R,E87V 12536 D36S,S116N 17406 K101F,R35I 22276
R37V,P121A 7667 L133R,E87D 12537 D36S,S116K 17407 K101F,R35L 22277
R37V,K10E 7668 L133R,R88L 12538 D36S,S116H 17408 K101F,R35M 22278
R37V,Y15A 7669 L133R,R88Y 12539 D36S,S116Q 17409 K101F,R35P 22279
R37V,Y15F 7670 L133R,R88D 12540 D36S,C117K 17410 K101F,R35S 22280
R37V,S17A 7671 L133R,L89A 12541 D36S,C117R 17411 K101F,R35V 22281
R37V,S17K 7672 L133R,E90K 12542 D36S,C117S 17412 K101F,D36A 22282
R37V,N18R 7673 L133R,E91A 12543 D36S,C117Q 17413 K101F,D36K 22283
R37V,H21A 7674 L133R,N92A 12544 D36S,C117T 17414 K101F,R37E 22284
R37V,L26A 7675 L133R,Y94V 12545 D36S,C117P 17415 K101F,R37A 22285
R37V,P27A 7676 L133R,Y94A 12546 D36S,C117A 17416 K101F,S38A 22286
R37V,D28K 7677 L133R,N95A 12547 D36S,C117I 17417 K101F,D39A 22287
D36S,K118
R37V,T30A 7678 L133R,N95V 12548 17418 K101F,Q40A 22288
W
R37V,T34A 7679 L133R,T96A 12549 D36S,K118H 17419 K101F,Q43A 22289
R37V,R35A 7680 L133R,K100E 12550 D36S,K118R 17420 K101F,Q43E 22290
R37V,R35E 7681 L133R,K101E 12551 D36S,K118S 17421 K101F,Q43K 22291
D36S,K118
R37V,R35F 7682 L133R,H102A 12552 17422 K101F,S47A 22292
M
R37V,R35G 7683 L133R,K105A 12553 D36S,K118L 17423 K101F,S47V 22293
R37V,R35I 7684 L133R,K105E 12554 D36S,P121A 17424 K101F,A48G 22294
R37V,R35L 7685 L133R,K113A 12555 D36S,R122A 17425 K101F,E53A 22295
R37V,R35M 7686 L133R,S116W 12556 D36S,R122E 17426 K101F,Y55A 22296
R37V,R35P 7687 L133R,S116Y 12557 D36S,H124A 17427 K101F,Y55F 22297
R37V,R35S 7688 L133R,S116F 12558 D36S,Q127A 17428 K101F,K57A 22298
R37V,R35V 7689 L133R,S116G 12559 D36S,Q127K 17429 K101F,K57E 22299
R37V,D36A 7690 L133R,S116N 12560 D36S,K128E 17430 K101F,E60A 22300
R37V,D36K 7691 L133R,S116K 12561 D36S,L133A 17431 K101F,E60K 22301
R37V,S38A 7692 L133R,S116H 12562 D36S,L133V 17432 K101F,T61A 22302
R37V,D39A 7693 L133R,S116Q 12563 D36T,K10E 17433 K101F,Q63A 22303
R37V,Q40A 7694 L133R,C117K 12564 D36T,Y15A 17434 K101F,Y64A 22304
R37V,Q43A 7695 L133R,C117R 12565 D36T,Y15F 17435 K101F,D68A 22305
R37V,Q43E 7696 L133R,C117S 12566 D36T,S17A 17436 K101F,T78A 22306
R37V,Q43K 7697 L133R,C117Q 12567 D36T,S17K 17437 K101F,P79A 22307
R37V,S47A 7698 L133R,C117T 12568 D36T,N18R 17438 K101F,E81K 22308 R37V,S47V 7699 L133R,C117P 12569 D36T,H21A 17439 K101F,L84A 22309
R37V,A48G 7700 L133R,C117A 12570 D36T,L26A 17440 K101F,E87K 22310
R37V,E53A 7701 L133R,C117I 12571 D36T,P27A 17441 K101F,E87Q 22311
L133R,K118
R37V,Y55A 7702 12572 D36T,D28K 17442 K101F,E87V 22312
W
R37V,Y55F 7703 L133R,K118H 12573 D36T,T30A 17443 K101F,E87D 22313
R37V,K57A 7704 L133R,K118R 12574 D36T,T34A 17444 K101F,R88L 22314
R37V,K57E 7705 L133R,K118S 12575 D36T,R35A 17445 K101F,R88Y 22315
L133R,K118
R37V,E60A 7706 12576 D36T,R35E 17446 K101F,R88D 22316
M
R37V,E60K 7707 L133R,K118L 12577 D36T,R35F 17447 K101F,L89A 22317
R37V,T61A 7708 L133R,P121A 12578 D36T,R35G 17448 K101F,E90K 22318
R37V,Q63A 7709 L133R,R122A 12579 D36T,R35I 17449 K101F,E91A 22319
R37V,Y64A 7710 L133R,R122E 12580 D36T,R35L 17450 K101F,N92A 22320
R37V,D68A 7711 L133R,H124A 12581 D36T,R35M 17451 K101F,Y94V 22321
R37V,T78A 7712 L133R,Q127A 12582 D36T,R35P 17452 K101F,Y94A 22322
R37V,P79A 7713 L133R,Q127K 12583 D36T,R35S 17453 K101F,N95A 22323
R37V,E81K 7714 L133R,K128E 12584 D36T,R35V 17454 K101F,N95V 22324
R37V,L84A 7715 L133S,K10E 12585 D36T,R37E 17455 K101F,T96A 22325
R37V,E87K 7716 L133S,Y15A 12586 D36T,R37A 17456 K101F,K100E 22326
R37V,E87Q 7717 L133S,Y15F 12587 D36T,S38A 17457 K101F,H102A 22327
R37V,E87V 7718 L133S,S17A 12588 D36T,D39A 17458 K101F,K105A 22328
R37V,E87D 7719 L133S,S17K 12589 D36T,Q40A 17459 K101F,K105E 22329
R37V,R88L 7720 L133S,N18R 12590 D36T,Q43A 17460 K101F,K113A 22330
R37V,R88Y 7721 L133S,H21A 12591 D36T,Q43E 17461 K101F,S116W 22331
R37V,R88D 7722 L133S,L26A 12592 D36T,Q43K 17462 K101F,S116Y 22332
R37V,L89A 7723 L133S,P27A 12593 D36T,S47A 17463 K101F,S116F 22333
R37V,E90K 7724 L133S,D28K 12594 D36T,S47V 17464 K101F,S116G 22334
R37V,E91A 7725 L133S,T30A 12595 D36T,A48G 17465 K101F,S116N 22335
R37V,N92A 7726 L133S,T34A 12596 D36T,E53A 17466 K101F,S116K 22336
R37V,Y94V 7727 L133S,R35A 12597 D36T,Y55A 17467 K101F,S116H 22337
R37V,Y94A 7728 L133S,R35E 12598 D36T,Y55F 17468 K101F,S116Q 22338
R37V,N95A 7729 L133S,R35F 12599 D36T,K57A 17469 K101F,C117K 22339
R37V,N95V 7730 L133S,R35G 12600 D36T,K57E 17470 K101F,C117R 22340
R37V,T96A 7731 L133S,R35I 12601 D36T,E60A 17471 K101F,C117S 22341
R37V,K100E 7732 L133S,R35L 12602 D36T,E60K 17472 K101F,C117Q 22342
R37V,K101E 7733 L133S,R35M 12603 D36T,T61A 17473 K101F,C117T 22343
R37V,H102A 7734 L133S,R35P 12604 D36T,Q63A 17474 K101F,C117P 22344
R37V,K105A 7735 L133S,R35S 12605 D36T,Y64A 17475 K101F,C117A 22345
R37V,K105E 7736 L133S,R35V 12606 D36T,D68A 17476 K101F,C117I 22346
K101F,K118
R37V,K113A 7737 L133S,D36A 12607 D36T,T78A 17477 22347
W
R37V,S116W 7738 L133S,D36K 12608 D36T,P79A 17478 K101F,K118H 22348
R37V,S116Y 7739 L133S,R37E 12609 D36T,E81K 17479 K101F,K118R 22349
R37V,S116F 7740 L133S,R37A 12610 D36T,L84A 17480 K101F,K118S 22350
K101F,K118
R37V,S116G 7741 L133S,S38A 12611 D36T,E87K 17481 22351
M
R37V,S116N 7742 L133S,D39A 12612 D36T,E87Q 17482 K101F,K118L 22352 R37V,S116K 7743 L133S,Q40A 12613 D36T,E87V 17483 K101F,P121A 22353
R37V,S116H 7744 L133S,Q43A 12614 D36T,E87D 17484 K101F,R122A 22354
R37V,S116Q 7745 L133S,Q43E 12615 D36T,R88L 17485 K101F,R122E 22355
R37V,C117K 7746 L133S,Q43K 12616 D36T,R88Y 17486 K101F,H124A 22356
R37V,C117R 7747 L133S,S47A 12617 D36T,R88D 17487 K101F,Q127A 22357
R37V,C117S 7748 L133S,S47V 12618 D36T,L89A 17488 K101F,Q127K 22358
R37V,C117Q 7749 L133S,A48G 12619 D36T,E90K 17489 K101F,K128E 22359
R37V,C117T 7750 L133S,E53A 12620 D36T,E91A 17490 K101F,L133A 22360
R37V,C117P 7751 L133S,Y55A 12621 D36T,N92A 17491 K101F,L133V 22361
R37V,C117A 7752 L133S,Y55F 12622 D36T,Y94V 17492 K101G,K10E 22362
R37V,C117I 7753 L133S,K57A 12623 D36T,Y94A 17493 K101G,Y15A 22363
R37V,K118
7754 L133S,K57E 12624 D36T,N95A 17494 K101G,Y15F 22364 W
R37V,K118H 7755 L133S,E60A 12625 D36T,N95V 17495 K101G,S17A 22365
R37V,K118R 7756 L133S,E60K 12626 D36T,T96A 17496 K101G,S17K 22366
R37V,K118S 7757 L133S,T61A 12627 D36T,K100E 17497 K101G,N18R 22367
R37V,K118
7758 L133S,Q63A 12628 D36T,K101E 17498 K101G,H21A 22368 M
R37V,K118L 7759 L133S,Y64A 12629 D36T,H102A 17499 K101G,L26A 22369
R37V,P121A 7760 L133S,D68A 12630 D36T,K105A 17500 K101G,P27A 22370
R37V,R122A 7761 L133S,T78A 12631 D36T,K105E 17501 K101G,D28K 22371
R37V,R122E 7762 L133S,P79A 12632 D36T,K113A 17502 K101G,T30A 22372
R37V,H124A 7763 L133S,E81K 12633 D36T,S116W 17503 K101G,T34A 22373
R37V,Q127A 7764 L133S,L84A 12634 D36T,S116Y 17504 K101G,R35A 22374
R37V,Q127K 7765 L133S,E87K 12635 D36T,S116F 17505 K101G,R35E 22375
R37V,K128E 7766 L133S,E87Q 12636 D36T,S116G 17506 K101G,R35F 22376
R37V,L133A 7767 L133S,E87V 12637 D36T,S116N 17507 K101G,R35G 22377
R37V,L133V 7768 L133S,E87D 12638 D36T,S116K 17508 K101G,R35I 22378
R37W,Q43D 7769 L133S,R88L 12639 D36T,S116H 17509 K101G,R35L 22379
R37W,Q43F 7770 L133S,R88Y 12640 D36T,S116Q 17510 K101G,R35M 22380
R37W,Q43G 7771 L133S,R88D 12641 D36T,C117K 17511 K101G,R35P 22381
R37W,Q43H 7772 L133S,L89A 12642 D36T,C117R 17512 K101G,R35S 22382
R37W,Q43I 7773 L133S,E90K 12643 D36T,C117S 17513 K101G,R35V 22383
R37W,Q43L 7774 L133S,E91A 12644 D36T,C117Q 17514 K101G,D36A 22384
R37W,Q43M 7775 L133S,N92A 12645 D36T,C117T 17515 K101G,D36K 22385
R37W,Q43N 7776 L133S,Y94V 12646 D36T,C117P 17516 K101G,R37E 22386
R37W,Q43P 7777 L133S,Y94A 12647 D36T,C117A 17517 K101G,R37A 22387
R37W,Q43S 7778 L133S,N95A 12648 D36T,C117I 17518 K101G,S38A 22388
D36T,K118
R37W,Q43T 7779 L133S,N95V 12649 17519 K101G,D39A 22389
W
R37W,Q43V 7780 L133S,T96A 12650 D36T,K118H 17520 K101G,Q40A 22390
R37W,Q43W 7781 L133S,K100E 12651 D36T,K118R 17521 K101G,Q43A 22391
R37W,Q43Y 7782 L133S,K101E 12652 D36T,K118S 17522 K101G,Q43E 22392
D36T,K118
R37W,D28A 7783 L133S,H102A 12653 17523 K101G,Q43K 22393
M
R37W,E49A 7784 L133S,K105A 12654 D36T,K118L 17524 K101G,S47A 22394
R37W,P121A 7785 L133S,K105E 12655 D36T,P121A 17525 K101G,S47V 22395
R37W,K10E 7786 L133S,K113A 12656 D36T,R122A 17526 K101G,A48G 22396 R37W,Y15A 7787 L133S,S116W 12657 D36T,R122E 17527 K101G,E53A 22397
R37W,Y15F 7788 L133S,S116Y 12658 D36T,H124A 17528 K101G,Y55A 22398
R37W,S17A 7789 L133S,S116F 12659 D36T,Q127A 17529 K101G,Y55F 22399
R37W,S17K 7790 L133S,S116G 12660 D36T,Q127K 17530 K101G,K57A 22400
R37W,N18R 7791 L133S,S116N 12661 D36T,K128E 17531 K101G,K57E 22401
R37W,H21A 7792 L133S,S116K 12662 D36T,L133A 17532 K101G,E60A 22402
R37W,L26A 7793 L133S,S116H 12663 D36T,L133V 17533 K101G,E60K 22403
R37W,P27A 7794 L133S,S116Q 12664 D36V,K10E 17534 K101G,T61A 22404
R37W,D28K 7795 L133S,C117K 12665 D36V,Y15A 17535 K101G,Q63A 22405
R37W,T30A 7796 L133S,C117R 12666 D36V,Y15F 17536 K101G,Y64A 22406
R37W,T34A 7797 L133S,C117S 12667 D36V,S17A 17537 K101G,D68A 22407
R37W,R35A 7798 L133S,C117Q 12668 D36V,S17K 17538 K101G,T78A 22408
R37W,R35E 7799 L133S,C117T 12669 D36V,N18R 17539 K101G,P79A 22409
R37W,R35F 7800 L133S,C117P 12670 D36V,H21A 17540 K101G,E81K 22410
R37W,R35G 7801 L133S,C117A 12671 D36V,L26A 17541 K101G,L84A 22411
R37W,R35I 7802 L133S,C117I 12672 D36V,P27A 17542 K101G,E87K 22412
L133S,K118
R37W,R35L 7803 12673 D36V,D28K 17543 K101G,E87Q 22413
W
R37W,R35M 7804 L133S,K118H 12674 D36V,T30A 17544 K101G,E87V 22414
R37W,R35P 7805 L133S,K118R 12675 D36V,T34A 17545 K101G,E87D 22415
R37W,R35S 7806 L133S,K118S 12676 D36V,R35A 17546 K101G,R88L 22416
R37W,R35V 7807 L133S,K118M 12677 D36V,R35E 17547 K101G,R88Y 22417
R37W,D36A 7808 L133S,K118L 12678 D36V,R35F 17548 K101G,R88D 22418
R37W,D36K 7809 L133S,P121A 12679 D36V,R35G 17549 K101G,L89A 22419
R37W,S38A 7810 L133S,R122A 12680 D36V,R35I 17550 K101G,E90K 22420
R37W,D39A 7811 L133S,R122E 12681 D36V,R35L 17551 K101G,E91A 22421
R37W,Q40A 7812 L133S,H124A 12682 D36V,R35M 17552 K101G,N92A 22422
R37W,Q43A 7813 L133S,Q127A 12683 D36V,R35P 17553 K101G,Y94V 22423
R37W,Q43E 7814 L133S,Q127K 12684 D36V,R35S 17554 K101G,Y94A 22424
R37W,Q43K 7815 L133S,K128E 12685 D36V,R35V 17555 K101G,N95A 22425
R37W,S47A 7816 L133T,K10E 12686 D36V,R37E 17556 K101G,N95V 22426
R37W,S47V 7817 L133T,Y15A 12687 D36V,R37A 17557 K101G,T96A 22427
R37W,A48G 7818 L133T,Y15F 12688 D36V,S38A 17558 K101G,K100E 22428
K101G,H102
R37W,E53A 7819 L133T,S17A 12689 D36V,D39A 17559 22429
A
K101G,K105
R37W,Y55A 7820 L133T,S17K 12690 D36V,Q40A 17560 22430
A
R37W,Y55F 7821 L133T,N18R 12691 D36V,Q43A 17561 K101G,K105E 22431
K101G,K113
R37W,K57A 7822 L133T,H21A 12692 D36V,Q43E 17562 22432
A
K101G,S116
R37W,K57E 7823 L133T,L26A 12693 D36V,Q43K 17563 22433
W
R37W,E60A 7824 L133T,P27A 12694 D36V,S47A 17564 K101G,S116Y 22434
R37W,E60K 7825 L133T,D28K 12695 D36V,S47V 17565 K101G,S116F 22435
R37W,T61A 7826 L133T,T30A 12696 D36V,A48G 17566 K101G,S116G 22436
R37W,Q63A 7827 L133T,T34A 12697 D36V,E53A 17567 K101G,S116N 22437
R37W,Y64A 7828 L133T,R35A 12698 D36V,Y55A 17568 K101G,S116K 22438
R37W,D68A 7829 L133T,R35E 12699 D36V,Y55F 17569 K101G,S116H 22439
Figure imgf000200_0001
Figure imgf000201_0001
Figure imgf000202_0001
Figure imgf000203_0001
Figure imgf000204_0001
L133W,K118
Q43D,P121A 8007 12877 D36Y,T34A 17747 K101I,E87Q 22617
R
L133W,K118
Q43D,K10E 8008 12878 D36Y,R35A 17748 K101I,E87V 22618
S
L133W,K118
Q43D,Y15A 8009 12879 D36Y,R35E 17749 K101I,E87D 22619
M
L133W,K118
Q43D,Y15F 8010 12880 D36Y,R35F 17750 K101I,R88L 22620
L
L133W,P121
Q43D,S17A 8011 12881 D36Y,R35G 17751 K101I,R88Y 22621
A
L133W,R122
Q43D,S17K 8012 12882 D36Y,R35I 17752 K101I,R88D 22622
A
L133W,R122
Q43D,N18R 8013 12883 D36Y,R35L 17753 K101I,L89A 22623
E
L133W,H124
Q43D,H21A 8014 12884 D36Y,R35M 17754 K101I,E90K 22624
A
L133W,Q127
Q43D,L26A 8015 12885 D36Y,R35P 17755 K101I,E91A 22625
A
L133W,Q127
Q43D,P27A 8016 12886 D36Y,R35S 17756 K101I,N92A 22626
K
L133W,K128
Q43D,D28K 8017 12887 D36Y,R35V 17757 K101I,Y94V 22627
E
Q43D,T30A 8018 L133Y,K10E 12888 D36Y,R37E 17758 K101I,Y94A 22628
Q43D,T34A 8019 L133Y,Y15A 12889 D36Y,R37A 17759 K101I,N95A 22629
Q43D,R35A 8020 L133Y,Y15F 12890 D36Y,S38A 17760 K101I,N95V 22630
Q43D,R35E 8021 L133Y,S17A 12891 D36Y,D39A 17761 K101I,T96A 22631
Q43D,R35F 8022 L133Y,S17K 12892 D36Y,Q40A 17762 ΚΙΟΠ,ΚΙΟΟΕ 22632
Q43D,R35G 8023 L133Y,N18R 12893 D36Y,Q43A 17763 K101I,H102A 22633
Q43D,R35I 8024 L133Y,H21A 12894 D36Y,Q43E 17764 K101I,K105A 22634
Q43D,R35L 8025 L133Y,L26A 12895 D36Y,Q43K 17765 K101I,K105E 22635
Q43D,R35M 8026 L133Y,P27A 12896 D36Y,S47A 17766 K101I,K113A 22636
Q43D,R35P 8027 L133Y,D28K 12897 D36Y,S47V 17767 K101I,S116W 22637
Q43D,R35S 8028 L133Y,T30A 12898 D36Y,A48G 17768 K101I,S116Y 22638
Q43D,R35V 8029 L133Y,T34A 12899 D36Y,E53A 17769 K101I,S116F 22639
Q43D,D36A 8030 L133Y,R35A 12900 D36Y,Y55A 17770 K101I,S116G 22640
Q43D,D36K 8031 L133Y,R35E 12901 D36Y,Y55F 17771 K101I,S116N 22641
Q43D,R37E 8032 L133Y,R35F 12902 D36Y,K57A 17772 K101I,S116K 22642
Q43D,R37A 8033 L133Y,R35G 12903 D36Y,K57E 17773 K101I,S116H 22643
Q43D,S38A 8034 L133Y,R35I 12904 D36Y,E60A 17774 K101I,S116Q 22644
Q43D,D39A 8035 L133Y,R35L 12905 D36Y,E60K 17775 K101I,C117K 22645
Q43D,Q40A 8036 L133Y,R35M 12906 D36Y,T61A 17776 K101I,C117R 22646
Q43D,S47A 8037 L133Y,R35P 12907 D36Y,Q63A 17777 K101I,C117S 22647
Q43D,S47V 8038 L133Y,R35S 12908 D36Y,Y64A 17778 K101I,C117Q 22648
Q43D,A48G 8039 L133Y,R35V 12909 D36Y,D68A 17779 K101I,C117T 22649
Q43D,E53A 8040 L133Y,D36A 12910 D36Y,T78A 17780 K101I,C117P 22650
Q43D,Y55A 8041 L133Y,D36K 12911 D36Y,P79A 17781 K101I,C117A 22651
Q43D,Y55F 8042 L133Y,R37E 12912 D36Y,E81K 17782 K101I,C117I 22652
Q43D,K57A 8043 L133Y,R37A 12913 D36Y,L84A 17783 K101I,K118W 22653
Q43D,K57E 8044 L133Y,S38A 12914 D36Y,E87K 17784 K101I,K118H 22654
Q43D,E60A 8045 L133Y,D39A 12915 D36Y,E87Q 17785 K101I,K118R 22655
Q43D,E60K 8046 L133Y,Q40A 12916 D36Y,E87V 17786 K101I,K118S 22656 Q43D,T61A 8047 L133Y,Q43A 12917 D36Y,E87D 17787 K101I,K118M 22657
Q43D,Q63A 8048 L133Y,Q43E 12918 D36Y,R88L 17788 K101I,K118L 22658
Q43D,Y64A 8049 L133Y,Q43K 12919 D36Y,R88Y 17789 K101I,P121A 22659
Q43D,D68A 8050 L133Y,S47A 12920 D36Y,R88D 17790 K101I,R122A 22660
Q43D,T78A 8051 L133Y,S47V 12921 D36Y,L89A 17791 K101I,R122E 22661
Q43D,P79A 8052 L133Y,A48G 12922 D36Y,E90K 17792 K101I,H124A 22662
Q43D,E81K 8053 L133Y,E53A 12923 D36Y,E91A 17793 K101I,Q127A 22663
Q43D,L84A 8054 L133Y,Y55A 12924 D36Y,N92A 17794 K101I,Q127K 22664
Q43D,E87K 8055 L133Y,Y55F 12925 D36Y,Y94V 17795 K101I,K128E 22665
Q43D,E87Q 8056 L133Y,K57A 12926 D36Y,Y94A 17796 K101I,L133A 22666
Q43D,E87V 8057 L133Y,K57E 12927 D36Y,N95A 17797 K101I,L133V 22667
Q43D,E87D 8058 L133Y,E60A 12928 D36Y,N95V 17798 K101L,K10E 22668
Q43D,R88L 8059 L133Y,E60K 12929 D36Y,T96A 17799 K101L,Y15A 22669
Q43D,R88Y 8060 L133Y,T61A 12930 D36Y,K100E 17800 K101L,Y15F 22670
Q43D,R88D 8061 L133Y,Q63A 12931 D36Y,K101E 17801 K101L,S17A 22671
D36Y,H102
Q43D,L89A 8062 L133Y,Y64A 12932 17802 K101L,S17K 22672
A
D36Y,K105
Q43D,E90K 8063 L133Y,D68A 12933 17803 K101L,N18R 22673
A
Q43D,E91A 8064 L133Y,T78A 12934 D36Y,K105E 17804 K101L,H21A 22674
D36Y,K113
Q43D,N92A 8065 L133Y,P79A 12935 17805 K101L,L26A 22675
A
D36Y,S116
Q43D,Y94V 8066 L133Y,E81K 12936 17806 K101L,P27A 22676
W
Q43D,Y94A 8067 L133Y,L84A 12937 D36Y,S116Y 17807 K101L,D28K 22677
Q43D,N95A 8068 L133Y,E87K 12938 D36Y,S116F 17808 K101L,T30A 22678
Q43D,N95V 8069 L133Y,E87Q 12939 D36Y,S116G 17809 K101L,T34A 22679
Q43D,T96A 8070 L133Y,E87V 12940 D36Y,S116N 17810 K101L,R35A 22680
Q43D,K100E 8071 L133Y,E87D 12941 D36Y,S116K 17811 K101L,R35E 22681
Q43D,K101E 8072 L133Y,R88L 12942 D36Y,S116H 17812 K101L,R35F 22682
Q43D,H102A 8073 L133Y,R88Y 12943 D36Y,S116Q 17813 K101L,R35G 22683
Q43D,K105A 8074 L133Y,R88D 12944 D36Y,C117K 17814 K101L,R35I 22684
Q43D,K105E 8075 L133Y,L89A 12945 D36Y,C117R 17815 K101L,R35L 22685
Q43D,K113A 8076 L133Y,E90K 12946 D36Y,C117S 17816 K101L,R35M 22686
Q43D,S116W 8077 L133Y,E91A 12947 D36Y,C117Q 17817 K101L,R35P 22687
Q43D,S116Y 8078 L133Y,N92A 12948 D36Y,C117T 17818 K101L,R35S 22688
Q43D,S116F 8079 L133Y,Y94V 12949 D36Y,C117P 17819 K101L,R35V 22689
Q43D,S116G 8080 L133Y,Y94A 12950 D36Y,C117A 17820 K101L,D36A 22690
Q43D,S116N 8081 L133Y,N95A 12951 D36Y,C117I 17821 K101L,D36K 22691
D36Y,K118
Q43D,S116K 8082 L133Y,N95V 12952 17822 K101L,R37E 22692
W
D36Y,K118
Q43D,S116H 8083 L133Y,T96A 12953 17823 K101L,R37A 22693
H
Q43D,S116Q 8084 L133Y,K100E 12954 D36Y,K118R 17824 K101L,S38A 22694
Q43D,C117K 8085 L133Y,K101E 12955 D36Y,K118S 17825 K101L,D39A 22695
D36Y,K118
Q43D,C117R 8086 L133Y,H102A 12956 17826 K101L,Q40A 22696
M
Q43D,C117S 8087 L133Y,K105A 12957 D36Y,K118L 17827 K101L,Q43A 22697
Q43D,C117Q 8088 L133Y,K105E 12958 D36Y,P121A 17828 K101L,Q43E 22698 Q43D,C117T 8089 L133Y,K113A 12959 D36Y,R122A 17829 K101L,Q43K 22699
L133Y,S116
Q43D,C117P 8090 12960 D36Y,R122E 17830 K101L,S47A 22700
W
D36Y,H124
Q43D,C117A 8091 L133Y,S116Y 12961 17831 K101L,S47V 22701
A
D36Y,Q127
Q43D,C117I 8092 L133Y,S116F 12962 17832 K101L,A48G 22702
A
Q43D,K118 D36Y,Q127
8093 L133Y,S116G 12963 17833 K101L,E53A 22703 W K
Q43D,K118H 8094 L133Y,S116N 12964 D36Y,K128E 17834 K101L,Y55A 22704
Q43D,K118R 8095 L133Y,S116K 12965 D36Y,L133A 17835 K101L,Y55F 22705
Q43D,K118S 8096 L133Y,S116H 12966 D36Y,L133V 17836 K101L,K57A 22706
Q43D,K118
8097 L133Y,S116Q 12967 E87F,K10E 17837 K101L,K57E 22707 M
Q43D,K118L 8098 L133Y,C117K 12968 E87F,Y15A 17838 K101L,E60A 22708
Q43D,P121A 8099 L133Y,C117R 12969 E87F,Y15F 17839 K101L,E60K 22709
Q43D,R122A 8100 L133Y,C117S 12970 E87F,S17A 17840 K101L,T61A 22710
Q43D,R122E 8101 L133Y,C117Q 12971 E87F,S17K 17841 K101L,Q63A 22711
Q43D,H124A 8102 L133Y,C117T 12972 E87F,N18R 17842 K101L,Y64A 22712
Q43D,Q127A 8103 L133Y,C117P 12973 E87F,H21A 17843 K101L,D68A 22713
Q43D,Q127K 8104 L133Y,C117A 12974 E87F,L26A 17844 K101L,T78A 22714
Q43D,K128E 8105 L133Y,C117I 12975 E87F,P27A 17845 K101L,P79A 22715
L133Y,K118
Q43D,L133A 8106 12976 E87F,D28K 17846 K101L,E81K 22716
W
Q43D,L133V 8107 L133Y,K118H 12977 E87F,T30A 17847 K101L,L84A 22717
Q43F,D28A 8108 L133Y,K118R 12978 E87F,T34A 17848 K101L,E87K 22718
Q43F,E49A 8109 L133Y,K118S 12979 E87F,R35A 17849 K101L,E87Q 22719
L133Y,K118
Q43F,P121A 8110 12980 E87F,R35E 17850 K101L,E87V 22720
M
Q43F,K10E 8111 L133Y,K118L 12981 E87F,R35F 17851 K101L,E87D 22721
Q43F,Y15A 8112 L133Y,P121A 12982 E87F,R35G 17852 K101L,R88L 22722
Q43F,Y15F 8113 L133Y,R122A 12983 E87F,R35I 17853 K101L,R88Y 22723
Q43F,S17A 8114 L133Y,R122E 12984 E87F,R35L 17854 K101L,R88D 22724
Q43F,S17K 8115 L133Y,H124A 12985 E87F,R35M 17855 K101L,L89A 22725
Q43F,N18R 8116 L133Y,Q127A 12986 E87F,R35P 17856 K101L,E90K 22726
Q43F,H21A 8117 L133Y,Q127K 12987 E87F,R35S 17857 K101L,E91A 22727
Q43F,L26A 8118 L133Y,K128E 12988 E87F,R35V 17858 K101L,N92A 22728
Q43F,P27A 8119 K105D,K10E 12989 E87F,D36A 17859 K101L,Y94V 22729
Q43F,D28K 8120 K105D,Y15A 12990 E87F,D36K 17860 K101L,Y94A 22730
Q43F,T30A 8121 K105D,Y15F 12991 E87F,R37E 17861 K101L,N95A 22731
Q43F,T34A 8122 K105D,S17A 12992 E87F,R37A 17862 K101L,N95V 22732
Q43F,R35A 8123 K105D,S17K 12993 E87F,S38A 17863 K101L,T96A 22733
Q43F,R35E 8124 K105D,N18R 12994 E87F,D39A 17864 K101L,K100E 22734
Q43F,R35F 8125 K105D,H21A 12995 E87F,Q40A 17865 K101L,H102A 22735
Q43F,R35G 8126 K105D,L26A 12996 E87F,Q43A 17866 K101L,K105A 22736
Q43F,R35I 8127 K105D,P27A 12997 E87F,Q43E 17867 K101L,K105E 22737
Q43F,R35L 8128 K105D,D28K 12998 E87F,Q43K 17868 K101L,K113A 22738
K101L,S116
Q43F,R35M 8129 K105D,T30A 12999 E87F,S47A 17869 22739
W
Q43F,R35P 8130 K105D,T34A 13000 E87F,S47V 17870 K101L,S116Y 22740 Q43F,R35S 8131 K105D,R35A 13001 E87F,A48G 17871 K101L,S116F 22741
Q43F,R35V 8132 K105D,R35E 13002 E87F,E53A 17872 K101L,S116G 22742
Q43F,D36A 8133 K105D,R35F 13003 E87F,Y55A 17873 K101L,S116N 22743
Q43F,D36K 8134 K105D,R35G 13004 E87F,Y55F 17874 K101L,S116K 22744
Q43F,R37E 8135 K105D,R35I 13005 E87F,K57A 17875 K101L,S116H 22745
Q43F,R37A 8136 K105D,R35L 13006 E87F,K57E 17876 K101L,S116Q 22746
Q43F,S38A 8137 K105D,R35M 13007 E87F,E60A 17877 K101L,C117K 22747
Q43F,D39A 8138 K105D,R35P 13008 E87F,E60K 17878 K101L,C117R 22748
Q43F,Q40A 8139 K105D,R35S 13009 E87F,T61A 17879 K101L,C117S 22749
Q43F,S47A 8140 K105D,R35V 13010 E87F,Q63A 17880 K101L,C117Q 22750
Q43F,S47V 8141 K105D,D36A 13011 E87F,Y64A 17881 K101L,C117T 22751
Q43F,A48G 8142 K105D,D36K 13012 E87F,D68A 17882 K101L,C117P 22752
Q43F,E53A 8143 K105D,R37E 13013 E87F,T78A 17883 K101L,C117A 22753
Q43F,Y55A 8144 K105D,R37A 13014 E87F,P79A 17884 K101L,C117I 22754
K101L,K118
Q43F,Y55F 8145 K105D,S38A 13015 E87F,E81K 17885 22755
W
Q43F,K57A 8146 K105D,D39A 13016 E87F,L84A 17886 K101L,K118H 22756
Q43F,K57E 8147 K105D,Q40A 13017 E87F,R88L 17887 K101L,K118R 22757
Q43F,E60A 8148 K105D,Q43A 13018 E87F,R88Y 17888 K101L,K118S 22758
K101L,K118
Q43F,E60K 8149 K105D,Q43E 13019 E87F,R88D 17889 22759
M
Q43F,T61A 8150 K105D,Q43K 13020 E87F,L89A 17890 K101L,K118L 22760
Q43F,Q63A 8151 K105D,S47A 13021 E87F,E90K 17891 K101L,P121A 22761
Q43F,Y64A 8152 K105D,S47V 13022 E87F,E91A 17892 K101L,R122A 22762
Q43F,D68A 8153 K105D,A48G 13023 E87F,N92A 17893 K101L,R122E 22763
Q43F,T78A 8154 K105D,E53A 13024 E87F,Y94V 17894 K101L,H124A 22764
Q43F,P79A 8155 K105D,Y55A 13025 E87F,Y94A 17895 K101L,Q127A 22765
Q43F,E81K 8156 K105D,Y55F 13026 E87F,N95A 17896 K101L,Q127K 22766
Q43F,L84A 8157 K105D,K57A 13027 E87F,N95V 17897 K101L,K128E 22767
Q43F,E87K 8158 K105D,K57E 13028 E87F,T96A 17898 K101L,L133A 22768
Q43F,E87Q 8159 K105D,E60A 13029 E87F,K100E 17899 K101L,L133V 22769
Q43F,E87V 8160 K105D,E60K 13030 E87F,K101E 17900 ΚΙΟΙΜ,ΚΙΟΕ 22770
Q43F,E87D 8161 K105D,T61A 13031 E87F,H102A 17901 K101M,Y15A 22771
Q43F,R88L 8162 K105D,Q63A 13032 E87F,K105A 17902 K101M,Y15F 22772
Q43F,R88Y 8163 K105D,Y64A 13033 E87F,K105E 17903 K101M,S17A 22773
Q43F,R88D 8164 K105D,D68A 13034 E87F,K113A 17904 K101M,S17K 22774
Q43F,L89A 8165 K105D,T78A 13035 E87F,S116W 17905 K101M,N18R 22775
Q43F,E90K 8166 K105D,P79A 13036 E87F,S116Y 17906 K101M,H21A 22776
Q43F,E91A 8167 K105D,E81K 13037 E87F,S116F 17907 K101M,L26A 22777
Q43F,N92A 8168 K105D,L84A 13038 E87F,S116G 17908 K101M,P27A 22778
Q43F,Y94V 8169 K105D,E87K 13039 E87F,S116N 17909 K101M,D28K 22779
Q43F,Y94A 8170 K105D,E87Q 13040 E87F,S116K 17910 K101M,T30A 22780
Q43F,N95A 8171 K105D,E87V 13041 E87F,S116H 17911 K101M/T34A 22781
Q43F,N95V 8172 K105D,E87D 13042 E87F,S116Q 17912 K101M,R35A 22782
Q43F,T96A 8173 K105D,R88L 13043 E87F,C117K 17913 K101M,R35E 22783
Q43F,K100E 8174 K105D,R88Y 13044 E87F,C117R 17914 K101M,R35F 22784
Q43F,K101E 8175 K105D,R88D 13045 E87F,C117S 17915 K101M,R35G 22785
Figure imgf000209_0001
Figure imgf000210_0001
H
K101M,K118
Q43G,K57A 8249 K105F,Q43A 13119 E87G,L89A 17989 22859
R
K101M,K118
Q43G,K57E 8250 K105F,Q43E 13120 E87G,E90K 17990 22860 s
K101M,K118
Q43G,E60A 8251 K105F,Q43K 13121 E87G,E91A 17991 22861
M
K101M,K118
Q43G,E60K 8252 K105F,S47A 13122 E87G,N92A 17992 22862
L
K101M,P121
Q43G,T61A 8253 K105F,S47V 13123 E87G,Y94V 17993 22863
A
K101M,R122
Q43G,Q63A 8254 K105F,A48G 13124 E87G,Y94A 17994 22864
A
K101M,R122
Q43G,Y64A 8255 K105F,E53A 13125 E87G,N95A 17995 22865
E
K101M,H124
Q43G,D68A 8256 K105F,Y55A 13126 E87G,N95V 17996 22866
A
K101M,Q127
Q43G,T78A 8257 K105F,Y55F 13127 E87G,T96A 17997 22867
A
K101M,Q127
Q43G,P79A 8258 K105F,K57A 13128 E87G,K100E 17998 22868
K
K101M,K128
Q43G,E81K 8259 K105F,K57E 13129 E87G,K101E 17999 22869
E
K101M,L133
Q43G,L84A 8260 K105F,E60A 13130 E87G,H102A 18000 22870
A
K101M,L133
Q43G,E87K 8261 K105F,E60K 13131 E87G,K105A 18001 22871
V
Q43G,E87Q 8262 K105F,T61A 13132 E87G,K105E 18002 ΚΙΟΙΝ,ΚΙΟΕ 22872
Q43G,E87V 8263 K105F,Q63A 13133 E87G,K113A 18003 K101N,Y15A 22873
Q43G,E87D 8264 K105F,Y64A 13134 E87G,S116W 18004 K101N,Y15F 22874
Q43G,R88L 8265 K105F,D68A 13135 E87G,S116Y 18005 K101N,S17A 22875
Q43G,R88Y 8266 K105F,T78A 13136 E87G,S116F 18006 K101N,S17K 22876
Q43G,R88D 8267 K105F,P79A 13137 E87G,S116G 18007 K101N,N18R 22877
Q43G,L89A 8268 K105F,E81K 13138 E87G,S116N 18008 K101N,H21A 22878
Q43G,E90K 8269 K105F,L84A 13139 E87G,S116K 18009 K101N,L26A 22879
Q43G,E91A 8270 K105F,E87K 13140 E87G,S116H 18010 K101N,P27A 22880
Q43G,N92A 8271 K105F,E87Q 13141 E87G,S116Q 18011 K101N,D28K 22881
Q43G,Y94V 8272 K105F,E87V 13142 E87G,C117K 18012 K101N,T30A 22882
Q43G,Y94A 8273 K105F,E87D 13143 E87G,C117R 18013 K101N,T34A 22883
Q43G,N95A 8274 K105F,R88L 13144 E87G,C117S 18014 K101N,R35A 22884
Q43G,N95V 8275 K105F,R88Y 13145 E87G,C117Q 18015 K101N,R35E 22885
Q43G,T96A 8276 K105F,R88D 13146 E87G,C117T 18016 K101N,R35F 22886
Q43G,K100E 8277 K105F,L89A 13147 E87G,C117P 18017 K101N,R35G 22887
Q43G,K101E 8278 K105F,E90K 13148 E87G,C117A 18018 K101N,R35I 22888
Q43G,H102A 8279 K105F,E91A 13149 E87G,C117I 18019 K101N,R35L 22889
E87G,K118
Q43G,K105A 8280 K105F,N92A 13150 18020 K101N,R35M 22890
W
Q43G,K105E 8281 K105F,Y94V 13151 E87G,K118H 18021 K101N,R35P 22891
Q43G,K113A 8282 K105F,Y94A 13152 E87G,K118R 18022 K101N,R35S 22892
Q43G,S116W 8283 K105F,N95A 13153 E87G,K118S 18023 K101N,R35V 22893
E87G,K118
Q43G,S116Y 8284 K105F,N95V 13154 18024 K101N,D36A 22894
M
Q43G,S116F 8285 K105F,T96A 13155 E87G,K118L 18025 K101N,D36K 22895 Q43G,S116G 8286 K105F,K100E 13156 E87G,P121A 18026 K101N,R37E 22896
Q43G,S116N 8287 K105F,K101E 13157 E87G,R122A 18027 K101N,R37A 22897
Q43G,S116K 8288 K105F,H102A 13158 E87G,R122E 18028 K101N,S38A 22898
Q43G,S116H 8289 K105F,K113A 13159 E87G,H124A 18029 K101N,D39A 22899
Q43G,S116Q 8290 K105F,S116W 13160 E87G,Q127A 18030 K101N,Q40A 22900
Q43G,C117K 8291 K105F,S116Y 13161 E87G,Q127K 18031 K101N,Q43A 22901
Q43G,C117R 8292 K105F,S116F 13162 E87G,K128E 18032 K101N,Q43E 22902
Q43G,C117S 8293 K105F,S116G 13163 E87G,L133A 18033 K101N,Q43K 22903
Q43G,C117Q 8294 K105F,S116N 13164 E87G,L133V 18034 K101N,S47A 22904
Q43G,C117T 8295 K105F,S116K 13165 E87H,K10E 18035 K101N,S47V 22905
Q43G,C117P 8296 K105F,S116H 13166 E87H,Y15A 18036 K101N,A48G 22906
Q43G,C117A 8297 K105F,S116Q 13167 E87H,Y15F 18037 K101N,E53A 22907
Q43G,C117I 8298 K105F,C117K 13168 E87H,S17A 18038 K101N,Y55A 22908
Q43G,K118
8299 K105F,C117R 13169 E87H,S17K 18039 K101N,Y55F 22909 W
Q43G,K118H 8300 K105F,C117S 13170 E87H,N18R 18040 K101N,K57A 22910
Q43G,K118R 8301 K105F,C117Q 13171 E87H,H21A 18041 K101N,K57E 22911
Q43G,K118S 8302 K105F,C117T 13172 E87H,L26A 18042 K101N,E60A 22912
Q43G,K118
8303 K105F,C117P 13173 E87H,P27A 18043 K101N,E60K 22913 M
Q43G,K118L 8304 K105F,C117A 13174 E87H,D28K 18044 K101N,T61A 22914
Q43G,P121A 8305 K105F,C117I 13175 E87H,T30A 18045 K101N,Q63A 22915
K105F,K118
Q43G,R122A 8306 13176 E87H,T34A 18046 K101N,Y64A 22916
W
Q43G,R122E 8307 K105F,K118H 13177 E87H,R35A 18047 K101N,D68A 22917
Q43G,H124A 8308 K105F,K118R 13178 E87H,R35E 18048 K101N,T78A 22918
Q43G,Q127A 8309 K105F,K118S 13179 E87H,R35F 18049 K101N,P79A 22919
K105F,K118
Q43G,Q127K 8310 13180 E87H,R35G 18050 K101N,E81K 22920
M
Q43G,K128E 8311 K105F,K118L 13181 E87H,R35I 18051 K101N,L84A 22921
Q43G,L133A 8312 K105F,P121A 13182 E87H,R35L 18052 K101N,E87K 22922
Q43G,L133V 8313 K105F,R122A 13183 E87H,R35M 18053 K101N,E87Q 22923
Q43H,D28A 8314 K105F,R122E 13184 E87H,R35P 18054 K101N,E87V 22924
Q43H,E49A 8315 K105F,H124A 13185 E87H,R35S 18055 K101N,E87D 22925
Q43H,P121A 8316 K105F,Q127A 13186 E87H,R35V 18056 K101N,R88L 22926
Q43H,K10E 8317 K105F,Q127K 13187 E87H,D36A 18057 K101N,R88Y 22927
Q43H,Y15A 8318 K105F,K128E 13188 E87H,D36K 18058 K101N,R88D 22928
Q43H,Y15F 8319 K105F,L133A 13189 E87H,R37E 18059 K101N,L89A 22929
Q43H,S17A 8320 K105F,L133V 13190 E87H,R37A 18060 K101N,E90K 22930
Q43H,S17K 8321 K105G,K10E 13191 E87H,S38A 18061 K101N,E91A 22931
Q43H,N18R 8322 K105G,Y15A 13192 E87H,D39A 18062 K101N,N92A 22932
Q43H,H21A 8323 K105G,Y15F 13193 E87H,Q40A 18063 K101N,Y94V 22933
Q43H,L26A 8324 K105G,S17A 13194 E87H,Q43A 18064 K101N,Y94A 22934
Q43H,P27A 8325 K105G,S17K 13195 E87H,Q43E 18065 K101N,N95A 22935
Q43H,D28K 8326 K105G,N18R 13196 E87H,Q43K 18066 K101N,N95V 22936
Q43H,T30A 8327 K105G,H21A 13197 E87H,S47A 18067 K101N,T96A 22937
Q43H,T34A 8328 K105G,L26A 13198 E87H,S47V 18068 K101N,K100E 22938
Q43H,R35A 8329 K105G,P27A 13199 E87H,A48G 18069 K101N,H102 22939
Figure imgf000213_0001
Figure imgf000214_0001
K105G,K118
Q43H,P121A 8408 13278 E87I,R35F 18148 K101P,Y64A 23018
H
K105G,K118
Q43H,R122A 8409 13279 E87I,R35G 18149 K101P,D68A 23019
R
Q43H,R122E 8410 K105G,K118S 13280 E87I,R35I 18150 K101P/T78A 23020
K105G,K118
Q43H,H124A 8411 13281 E87I,R35L 18151 K101P,P79A 23021
M
Q43H,Q127A 8412 K105G,K118L 13282 E87I,R35M 18152 K101P,E81K 23022
Q43H,Q127K 8413 K105G,P121A 13283 E87I,R35P 18153 K101P,L84A 23023
K105G,R122
Q43H,K128E 8414 13284 E87I,R35S 18154 K101P,E87K 23024
A
Q43H,L133A 8415 K105G,R122E 13285 E87I,R35V 18155 K101P,E87Q 23025
K105G,H124
Q43H,L133V 8416 13286 E87I,D36A 18156 K101P,E87V 23026
A
K105G,Q127
Q43I,D28A 8417 13287 E87I,D36K 18157 K101P,E87D 23027
A
K105G,Q127
Q43I,E49A 8418 13288 E87I,R37E 18158 K101P,R88L 23028
K
Q43I,P121A 8419 K105G,K128E 13289 E87I,R37A 18159 K101P,R88Y 23029
Q43I,K10E 8420 K105G,L133A 13290 E87I,S38A 18160 K101P,R88D 23030
Q43I,Y15A 8421 K105G,L133V 13291 E87I,D39A 18161 K101P,L89A 23031
Q43I,Y15F 8422 K105H,K10E 13292 E87I,Q40A 18162 K101P,E90K 23032
Q43I,S17A 8423 K105H,Y15A 13293 E87I,Q43A 18163 K101P,E91A 23033
Q43I,S17K 8424 K105H,Y15F 13294 E87I,Q43E 18164 K101P,N92A 23034
Q43I,N18R 8425 K105H,S17A 13295 E87I,Q43K 18165 K101P,Y94V 23035
Q43I,H21A 8426 K105H,S17K 13296 E87I,S47A 18166 K101P,Y94A 23036
Q43I,L26A 8427 K105H,N18R 13297 E87I,S47V 18167 K101P,N95A 23037
Q43I,P27A 8428 K105H,H21A 13298 E87I,A48G 18168 K101P,N95V 23038
Q43I,D28K 8429 K105H,L26A 13299 E87I,E53A 18169 K101P,T96A 23039
Q43I,T30A 8430 K105H,P27A 13300 E87I,Y55A 18170 K101P,K100E 23040
Q43I,T34A 8431 K105H,D28K 13301 E87I,Y55F 18171 K101P,H102A 23041
Q43I,R35A 8432 K105H,T30A 13302 E87I,K57A 18172 K101P,K105A 23042
Q43I,R35E 8433 K105H,T34A 13303 E87I,K57E 18173 K101P,K105E 23043
Q43I,R35F 8434 K105H,R35A 13304 E87I,E60A 18174 K101P,K113A 23044
Q43I,R35G 8435 K105H,R35E 13305 E87I,E60K 18175 K101P,S116W 23045
Q43I,R35I 8436 K105H,R35F 13306 E87I,T61A 18176 K101P,S116Y 23046
Q43I,R35L 8437 K105H,R35G 13307 E87I,Q63A 18177 K101P,S116F 23047
Q43I,R35M 8438 K105H,R35I 13308 E87I,Y64A 18178 K101P,S116G 23048
Q43I,R35P 8439 K105H,R35L 13309 E87I,D68A 18179 K101P,S116N 23049
Q43I,R35S 8440 K105H,R35M 13310 E87I,T78A 18180 K101P,S116K 23050
Q43I,R35V 8441 K105H,R35P 13311 E87I,P79A 18181 K101P,S116H 23051
Q43I,D36A 8442 K105H,R35S 13312 E87I,E81K 18182 K101P,S116Q 23052
Q43I,D36K 8443 K105H,R35V 13313 E87I,L84A 18183 K101P,C117K 23053
Q43I,R37E 8444 K105H,D36A 13314 E87I,R88L 18184 K101P,C117R 23054
Q43I,R37A 8445 K105H,D36K 13315 E87I,R88Y 18185 K101P,C117S 23055
Q43I,S38A 8446 K105H,R37E 13316 E87I,R88D 18186 K101P,C117Q 23056
Q43I,D39A 8447 K105H,R37A 13317 E87I,L89A 18187 K101P,C117T 23057
Q43I,Q40A 8448 K105H,S38A 13318 E87I,E90K 18188 K101P,C117P 23058
Q43I,S47A 8449 K105H,D39A 13319 E87I,E91A 18189 K101P,C117A 23059 Q43I,S47V 8450 K105H,Q40A 13320 E87I,N92A 18190 K101P,C117I 23060
K101P,K118
Q43I,A48G 8451 K105H,Q43A 13321 E87I,Y94V 18191 23061
W
Q43I,E53A 8452 K105H,Q43E 13322 E87I,Y94A 18192 K101P,K118H 23062
Q43I,Y55A 8453 K105H,Q43K 13323 E87I,N95A 18193 K101P,K118R 23063
Q43I,Y55F 8454 K105H,S47A 13324 E87I,N95V 18194 K101P,K118S 23064
K101P,K118
Q43I,K57A 8455 K105H,S47V 13325 E87I,T96A 18195 23065
M
Q43I,K57E 8456 K105H,A48G 13326 E87I,K100E 18196 K101P,K118L 23066
Q43I,E60A 8457 K105H,E53A 13327 E87I,K101E 18197 K101P,P121A 23067
Q43I,E60K 8458 K105H,Y55A 13328 E87I,H102A 18198 K101P,R122A 23068
Q43I,T61A 8459 K105H,Y55F 13329 E87I,K105A 18199 K101P,R122E 23069
Q43I,Q63A 8460 K105H,K57A 13330 E87I,K105E 18200 K101P,H124A 23070
Q43I,Y64A 8461 K105H,K57E 13331 E87I,K113A 18201 K101P,Q127A 23071
Q43I,D68A 8462 K105H,E60A 13332 E87I,S116W 18202 K101P,Q127K 23072
Q43I,T78A 8463 K105H,E60K 13333 E87I,S116Y 18203 K101P,K128E 23073
Q43I,P79A 8464 K105H,T61A 13334 E87I,S116F 18204 K101P,L133A 23074
Q43I,E81K 8465 K105H,Q63A 13335 E87I,S116G 18205 K101P,L133V 23075
Q43I,L84A 8466 K105H,Y64A 13336 E87I,S116N 18206 K101Q,K10E 23076
Q43I,E87K 8467 K105H,D68A 13337 E87I,S116K 18207 K101Q,Y15A 23077
Q43I,E87Q 8468 K105H,T78A 13338 E87I,S116H 18208 K101Q,Y15F 23078
Q43I,E87V 8469 K105H,P79A 13339 E87I,S116Q 18209 K101Q,S17A 23079
Q43I,E87D 8470 K105H,E81K 13340 E87I,C117K 18210 K101Q,S17K 23080
Q43I,R88L 8471 K105H,L84A 13341 E87I,C117R 18211 K101Q,N18R 23081
Q43I,R88Y 8472 K105H,E87K 13342 E87I,C117S 18212 K101Q,H21A 23082
Q43I,R88D 8473 K105H,E87Q 13343 E87I,C117Q 18213 K101Q,L26A 23083
Q43I,L89A 8474 K105H,E87V 13344 E87I,C117T 18214 K101Q,P27A 23084
Q43I,E90K 8475 K105H,E87D 13345 E87I,C117P 18215 K101Q,D28K 23085
Q43I,E91A 8476 K105H,R88L 13346 E87I,C117A 18216 K101Q,T30A 23086
Q43I,N92A 8477 K105H,R88Y 13347 E87I,C117I 18217 K101Q,T34A 23087
Q43I,Y94V 8478 K105H,R88D 13348 E87I,K118W 18218 K101Q,R35A 23088
Q43I,Y94A 8479 K105H,L89A 13349 E87I,K118H 18219 K101Q,R35E 23089
Q43I,N95A 8480 K105H,E90K 13350 E87I,K118R 18220 K101Q,R35F 23090
Q43I,N95V 8481 K105H,E91A 13351 E87I,K118S 18221 K101Q,R35G 23091
Q43I,T96A 8482 K105H,N92A 13352 E87I,K118M 18222 K101Q,R35I 23092
Q43I,K100E 8483 K105H,Y94V 13353 E87I,K118L 18223 K101Q,R35L 23093
Q43I,K101E 8484 K105H,Y94A 13354 E87I,P121A 18224 K101Q,R35M 23094
Q43I,H102A 8485 K105H,N95A 13355 E87I,R122A 18225 K101Q,R35P 23095
Q43I,K105A 8486 K105H,N95V 13356 E87I,R122E 18226 K101Q,R35S 23096
Q43I,K105E 8487 K105H,T96A 13357 E87I,H124A 18227 K101Q,R35V 23097
Q43I,K113A 8488 K105H,K100E 13358 E87I,Q127A 18228 K101Q,D36A 23098
Q43I,S116W 8489 K105H,K101E 13359 E87I,Q127K 18229 K101Q,D36K 23099
K105H,H102
Q43I,S116Y 8490 13360 E87I,K128E 18230 K101Q,R37E 23100
A
K105H,K113
Q43I,S116F 8491 13361 E87I,L133A 18231 K101Q,R37A 23101
A
K105H,S116
Q43I,S116G 8492 13362 E87I,L133V 18232 K101Q,S38A 23102
W
Figure imgf000217_0001
Figure imgf000218_0001
Q43L,E87Q 8571 K105I,E81K 13441 E87L,C117S 18311 K101R,S17A 23181
Q43L,E87V 8572 K105I,L84A 13442 E87L,C117Q 18312 K101R,S17K 23182
Q43L,E87D 8573 K105I,E87K 13443 E87L,C117T 18313 K101R,N18R 23183
Q43L,R88L 8574 K105I,E87Q 13444 E87L,C117P 18314 K101R,H21A 23184
Q43L,R88Y 8575 K105I,E87V 13445 E87L,C117A 18315 K101R,L26A 23185
Q43L,R88D 8576 K105I,E87D 13446 E87L,C117I 18316 K101R,P27A 23186
E87L,K118
Q43L,L89A 8577 K105I,R88L 13447 18317 K101R,D28K 23187
W
Q43L,E90K 8578 K105I,R88Y 13448 E87L,K118H 18318 K101R,T30A 23188
Q43L,E91A 8579 K105I,R88D 13449 E87L,K118R 18319 K101R,T34A 23189
Q43L,N92A 8580 K105I,L89A 13450 E87L,K118S 18320 K101R,R35A 23190
Q43L,Y94V 8581 K105I,E90K 13451 E87L,K118M 18321 K101R,R35E 23191
Q43L,Y94A 8582 K105I,E91A 13452 E87L,K118L 18322 K101R,R35F 23192
Q43L,N95A 8583 K105I,N92A 13453 E87L,P121A 18323 K101R,R35G 23193
Q43L,N95V 8584 K105I,Y94V 13454 E87L,R122A 18324 K101R,R35I 23194
Q43L,T96A 8585 K105I,Y94A 13455 E87L,R122E 18325 K101R,R35L 23195
Q43L,K100E 8586 K105I,N95A 13456 E87L,H124A 18326 K101R,R35M 23196
Q43L,K101E 8587 K105I,N95V 13457 E87L,Q127A 18327 K101R,R35P 23197
Q43L,H102A 8588 K105I,T96A 13458 E87L,Q127K 18328 K101R,R35S 23198
Q43L,K105A 8589 K105I,K100E 13459 E87L,K128E 18329 K101R,R35V 23199
Q43L,K105E 8590 K105I,K101E 13460 E87L,L133A 18330 K101R,D36A 23200
Q43L,K113A 8591 K105I,H102A 13461 E87L,L133V 18331 K101R,D36K 23201
Q43L,S116W 8592 K105I,K113A 13462 E87M,K10E 18332 K101R,R37E 23202
Q43L,S116Y 8593 K105I,S116W 13463 E87M,Y15A 18333 K101R,R37A 23203
Q43L,S116F 8594 K105I,S116Y 13464 E87M,Y15F 18334 K101R,S38A 23204
Q43L,S116G 8595 K105I,S116F 13465 E87M,S17A 18335 K101R,D39A 23205
Q43L,S116N 8596 K105I,S116G 13466 E87M,S17K 18336 K101R,Q40A 23206
Q43L,S116K 8597 K105I,S116N 13467 E87M,N18R 18337 K101R,Q43A 23207
Q43L,S116H 8598 K105I,S116K 13468 E87M,H21A 18338 K101R,Q43E 23208
Q43L,S116Q 8599 K105I,S116H 13469 E87M,L26A 18339 K101R,Q43K 23209
Q43L,C117K 8600 K105I,S116Q 13470 E87M,P27A 18340 K101R,S47A 23210
Q43L,C117R 8601 K105I,C117K 13471 E87M,D28K 18341 K101R,S47V 23211
Q43L,C117S 8602 K105I,C117R 13472 E87M,T30A 18342 K101R,A48G 23212
Q43L,C117Q 8603 K105I,C117S 13473 E87M,T34A 18343 K101R,E53A 23213
Q43L,C117T 8604 K105I,C117Q 13474 E87M,R35A 18344 K101R,Y55A 23214
Q43L,C117P 8605 K105I,C117T 13475 E87M,R35E 18345 K101R,Y55F 23215
Q43L,C117A 8606 K105I,C117P 13476 E87M,R35F 18346 K101R,K57A 23216
Q43L,C117I 8607 K105I,C117A 13477 E87M,R35G 18347 K101R,K57E 23217
Q43L,K118
8608 K105I,C117I 13478 E87M,R35I 18348 K101R,E60A 23218 W
Q43L,K118H 8609 K105I,K118W 13479 E87M,R35L 18349 K101R,E60K 23219
Q43L,K118R 8610 K105I,K118H 13480 E87M,R35M 18350 K101R,T61A 23220
Q43L,K118S 8611 K105I,K118R 13481 E87M,R35P 18351 K101R,Q63A 23221
Q43L,K118M 8612 K105I,K118S 13482 E87M,R35S 18352 K101R,Y64A 23222
Q43L,K118L 8613 K105I,K118M 13483 E87M,R35V 18353 K101R,D68A 23223
Q43L,P121A 8614 K105I,K118L 13484 E87M,D36A 18354 K101R,T78A 23224
Q43L,R122A 8615 K105I,P121A 13485 E87M,D36K 18355 K101R,P79A 23225 Q43L,R122E 8616 K105I,R122A 13486 E87M,R37E 18356 K101R,E81K 23226
Q43L,H124A 8617 K105I,R122E 13487 E87M,R37A 18357 K101R,L84A 23227
Q43L,Q127A 8618 K105I,H124A 13488 E87M,S38A 18358 K101R,E87K 23228
Q43L,Q127K 8619 K105I,Q127A 13489 E87M,D39A 18359 K101R,E87Q 23229
Q43L,K128E 8620 K105I,Q127K 13490 E87M,Q40A 18360 K101R,E87V 23230
Q43L,L133A 8621 K105I,K128E 13491 E87M,Q43A 18361 K101R,E87D 23231
Q43L,L133V 8622 K105I,L133A 13492 E87M,Q43E 18362 K101R,R88L 23232
Q43M,D28A 8623 K105I,L133V 13493 E87M,Q43K 18363 K101R,R88Y 23233
Q43M,E49A 8624 K105L,K10E 13494 E87M,S47A 18364 K101R,R88D 23234
Q43M,P121A 8625 K105L,Y15A 13495 E87M,S47V 18365 K101R,L89A 23235
Q43M,K10E 8626 K105L,Y15F 13496 E87M,A48G 18366 K101R,E90K 23236
Q43M,Y15A 8627 K105L,S17A 13497 E87M,E53A 18367 K101R,E91A 23237
Q43M,Y15F 8628 K105L,S17K 13498 E87M,Y55A 18368 K101R,N92A 23238
Q43M,S17A 8629 K105L,N18R 13499 E87M,Y55F 18369 K101R,Y94V 23239
Q43M,S17K 8630 K105L,H21A 13500 E87M,K57A 18370 K101R,Y94A 23240
Q43M,N18R 8631 K105L,L26A 13501 E87M,K57E 18371 K101R,N95A 23241
Q43M,H21A 8632 K105L,P27A 13502 E87M,E60A 18372 K101R,N95V 23242
Q43M,L26A 8633 K105L,D28K 13503 E87M,E60K 18373 K101R,T96A 23243
Q43M,P27A 8634 K105L,T30A 13504 E87M,T61A 18374 K101R,K100E 23244
K101R,H102
Q43M,D28K 8635 K105L,T34A 13505 E87M,Q63A 18375 23245
A
K101R,K105
Q43M,T30A 8636 K105L,R35A 13506 E87M,Y64A 18376 23246
A
Q43M,T34A 8637 K105L,R35E 13507 E87M,D68A 18377 K101R,K105E 23247
K101R,K113
Q43M,R35A 8638 K105L,R35F 13508 E87M,T78A 18378 23248
A
K101R,S116
Q43M,R35E 8639 K105L,R35G 13509 E87M,P79A 18379 23249
W
Q43M,R35F 8640 K105L,R35I 13510 E87M,E81K 18380 K101R,S116Y 23250
Q43M,R35G 8641 K105L,R35L 13511 E87M,L84A 18381 K101R,S116F 23251
Q43M,R35I 8642 K105L,R35M 13512 E87M,R88L 18382 K101R,S116G 23252
Q43M,R35L 8643 K105L,R35P 13513 E87M,R88Y 18383 K101R,S116N 23253
Q43M,R35M 8644 K105L,R35S 13514 E87M,R88D 18384 K101R,S116K 23254
Q43M,R35P 8645 K105L,R35V 13515 E87M,L89A 18385 K101R,S116H 23255
Q43M,R35S 8646 K105L,D36A 13516 E87M,E90K 18386 K101R,S116Q 23256
Q43M,R35V 8647 K105L,D36K 13517 E87M,E91A 18387 K101R,C117K 23257
Q43M,D36A 8648 K105L,R37E 13518 E87M,N92A 18388 K101R,C117R 23258
Q43M,D36K 8649 K105L,R37A 13519 E87M,Y94V 18389 K101R,C117S 23259
Q43M,R37E 8650 K105L,S38A 13520 E87M,Y94A 18390 K101R,C117Q 23260
Q43M,R37A 8651 K105L,D39A 13521 E87M,N95A 18391 K101R,C117T 23261
Q43M,S38A 8652 K105L,Q40A 13522 E87M,N95V 18392 K101R,C117P 23262
Q43M,D39A 8653 K105L,Q43A 13523 E87M,T96A 18393 K101R,C117A 23263
Q43M,Q40A 8654 K105L,Q43E 13524 E87M,K100E 18394 K101R,C117I 23264
K101R,K118
Q43M,S47A 8655 K105L,Q43K 13525 E87M,K101E 18395 23265
W
E87M,H102 K101R,K118
Q43M,S47V 8656 K105L,S47A 13526 18396 23266
A H
E87M,K105
Q43M,A48G 8657 K105L,S47V 13527 18397 K101R,K118R 23267
A
Figure imgf000221_0001
Figure imgf000222_0001
Q43N,Y15F 8731 K105M,H21A 13601 E87N,E60A 18471 K101S,Y94V 23341
Q43N,S17A 8732 K105M,L26A 13602 E87N,E60K 18472 K101S,Y94A 23342
Q43N,S17K 8733 K105M,P27A 13603 E87N,T61A 18473 K101S,N95A 23343
Q43N,N18R 8734 K105M,D28K 13604 E87N,Q63A 18474 K101S,N95V 23344
Q43N,H21A 8735 Κ105Μ/Γ30Α 13605 E87N,Y64A 18475 K101S,T96A 23345
Q43N,L26A 8736 K105M,T34A 13606 E87N,D68A 18476 K101S,K100E 23346
Q43N,P27A 8737 K105M,R35A 13607 E87N,T78A 18477 K101S,H102A 23347
Q43N,D28K 8738 K105M,R35E 13608 E87N,P79A 18478 K101S,K105A 23348
Q43N,T30A 8739 K105M,R35F 13609 E87N,E81K 18479 K101S,K105E 23349
Q43N,T34A 8740 K105M,R35G 13610 E87N,L84A 18480 K101S,K113A 23350
Q43N,R35A 8741 K105M,R35I 13611 E87N,R88L 18481 K101S,S116W 23351
Q43N,R35E 8742 K105M,R35L 13612 E87N,R88Y 18482 K101S,S116Y 23352
Q43N,R35F 8743 K105M,R35M 13613 E87N,R88D 18483 K101S,S116F 23353
Q43N,R35G 8744 K105M,R35P 13614 E87N,L89A 18484 K101S,S116G 23354
Q43N,R35I 8745 K105M,R35S 13615 E87N,E90K 18485 K101S,S116N 23355
Q43N,R35L 8746 K105M,R35V 13616 E87N,E91A 18486 K101S,S116K 23356
Q43N,R35M 8747 K105M,D36A 13617 E87N,N92A 18487 K101S,S116H 23357
Q43N,R35P 8748 K105M,D36K 13618 E87N,Y94V 18488 K101S,S116Q 23358
Q43N,R35S 8749 K105M,R37E 13619 E87N,Y94A 18489 K101S,C117K 23359
Q43N,R35V 8750 K105M,R37A 13620 E87N,N95A 18490 K101S,C117R 23360
Q43N,D36A 8751 K105M,S38A 13621 E87N,N95V 18491 K101S,C117S 23361
Q43N,D36K 8752 K105M,D39A 13622 E87N,T96A 18492 K101S,C117Q 23362
Q43N,R37E 8753 K105M,Q40A 13623 E87N,K100E 18493 K101S,C117T 23363
Q43N,R37A 8754 K105M,Q43A 13624 E87N,K101E 18494 K101S,C117P 23364
Q43N,S38A 8755 K105M,Q43E 13625 E87N,H102A 18495 K101S,C117A 23365
Q43N,D39A 8756 K105M,Q43K 13626 E87N,K105A 18496 K101S,C117I 23366
K101S,K118
Q43N,Q40A 8757 K105M,S47A 13627 E87N,K105E 18497 23367
W
Q43N,S47A 8758 K105M,S47V 13628 E87N,K113A 18498 K101S,K118H 23368
Q43N,S47V 8759 K105M,A48G 13629 E87N,S116W 18499 K101S,K118R 23369
Q43N,A48G 8760 K105M,E53A 13630 E87N,S116Y 18500 K101S,K118S 23370
K101S,K118
Q43N,E53A 8761 K105M,Y55A 13631 E87N,S116F 18501 23371
M
Q43N,Y55A 8762 K105M,Y55F 13632 E87N,S116G 18502 K101S,K118L 23372
Q43N,Y55F 8763 K105M,K57A 13633 E87N,S116N 18503 K101S,P121A 23373
Q43N,K57A 8764 K105M,K57E 13634 E87N,S116K 18504 K101S,R122A 23374
Q43N,K57E 8765 K105M,E60A 13635 E87N,S116H 18505 K101S,R122E 23375
Q43N,E60A 8766 K105M,E60K 13636 E87N,S116Q 18506 K101S,H124A 23376
Q43N,E60K 8767 K105M,T61A 13637 E87N,C117K 18507 K101S,Q127A 23377
Q43N,T61A 8768 K105M,Q63A 13638 E87N,C117R 18508 K101S,Q127K 23378
Q43N,Q63A 8769 K105M,Y64A 13639 E87N,C117S 18509 K101S,K128E 23379
Q43N,Y64A 8770 K105M,D68A 13640 E87N,C117Q 18510 K101S,L133A 23380
Q43N,D68A 8771 K105M,T78A 13641 E87N,C117T 18511 K101S,L133V 23381
Q43N,T78A 8772 K105M,P79A 13642 E87N,C117P 18512 ΚΙΟΙΤ,ΚΙΟΕ 23382
Q43N,P79A 8773 K105M,E81K 13643 E87N,C117A 18513 K101T,Y15A 23383
Q43N,E81K 8774 K105M,L84A 13644 E87N,C117I 18514 K101T,Y15F 23384
E87N,K118
Q43N,L84A 8775 K105M,E87K 13645 18515 K101T,S17A 23385
W
Figure imgf000224_0001
K105MK118
Q43N,C117P 8811 13681 E87P,R35V 18551 K101T,K57E 23421
W
K105MK118
Q43N,C117A 8812 13682 E87P,D36A 18552 K101T,E60A 23422
H
K105MK118
Q43N,C117I 8813 13683 E87P,D36K 18553 K101T,E60K 23423
R
Q43N,K118 K105MK118
8814 13684 E87P,R37E 18554 Κ101Τ/Γ61Α 23424 W S
K105MK118
Q43N,K118H 8815 13685 E87P,R37A 18555 K101T,Q63A 23425
M
K105MK118
Q43N,K118R 8816 13686 E87P,S38A 18556 K101T,Y64A 23426
L
K105MP121
Q43N,K118S 8817 13687 E87P,D39A 18557 K101T,D68A 23427
A
Q43N,K118 K105MR122
8818 13688 E87P,Q40A 18558 K101T/T78A 23428 M A
K105MR122
Q43N,K118L 8819 13689 E87P,Q43A 18559 K101T,P79A 23429
E
K105MH124
Q43N,P121A 8820 13690 E87P,Q43E 18560 K101T,E81K 23430
A
K105M,Q127
Q43N,R122A 8821 13691 E87P,Q43K 18561 K101T,L84A 23431
A
K105M,Q127
Q43N,R122E 8822 13692 E87P,S47A 18562 K101T,E87K 23432
K
K105M,K128
Q43N,H124A 8823 13693 E87P,S47V 18563 K101T,E87Q 23433
E
K105ML133
Q43N,Q127A 8824 13694 E87P,A48G 18564 K101T,E87V 23434
A
K105M,L133
Q43N,Q127K 8825 13695 E87P,E53A 18565 K101T,E87D 23435
V
Q43N,K128E 8826 K105N,K10E 13696 E87P,Y55A 18566 K101T,R88L 23436
Q43N,L133A 8827 K105N,Y15A 13697 E87P,Y55F 18567 K101T,R88Y 23437
Q43N,L133V 8828 K105N,Y15F 13698 E87P,K57A 18568 K101T,R88D 23438
Q43P,D28A 8829 K105N,S17A 13699 E87P,K57E 18569 K101T,L89A 23439
Q43P,E49A 8830 K105N,S17K 13700 E87P,E60A 18570 K101T,E90K 23440
Q43P,P121A 8831 K105N,N18R 13701 E87P,E60K 18571 K101T,E91A 23441
Q43P,K10E 8832 K105N,H21A 13702 E87P,T61A 18572 K101T,N92A 23442
Q43P,Y15A 8833 K105N,L26A 13703 E87P,Q63A 18573 K101T,Y94V 23443
Q43P,Y15F 8834 K105N,P27A 13704 E87P,Y64A 18574 K101T,Y94A 23444
Q43P,S17A 8835 K105N,D28K 13705 E87P,D68A 18575 K101T,N95A 23445
Q43P,S17K 8836 K105N,T30A 13706 E87P,T78A 18576 K101T,N95V 23446
Q43P,N18R 8837 Κ105Ν/Γ34Α 13707 E87P,P79A 18577 K101T,T96A 23447
Q43P,H21A 8838 K105N,R35A 13708 E87P,E81K 18578 K101T,K100E 23448
Q43P,L26A 8839 K105N,R35E 13709 E87P,L84A 18579 K101T,H102A 23449
Q43P,P27A 8840 K105N,R35F 13710 E87P,R88L 18580 K101T,K105A 23450
Q43P,D28K 8841 K105N,R35G 13711 E87P,R88Y 18581 K101T,K105E 23451
Q43P,T30A 8842 K105N,R35I 13712 E87P,R88D 18582 K101T,K113A 23452
K101T,S116
Q43P,T34A 8843 K105N,R35L 13713 E87P,L89A 18583 23453
W
Q43P,R35A 8844 K105N,R35M 13714 E87P,E90K 18584 K101T,S116Y 23454
Q43P,R35E 8845 K105N,R35P 13715 E87P,E91A 18585 K101T,S116F 23455
Q43P,R35F 8846 K105N,R35S 13716 E87P,N92A 18586 K101T,S116G 23456
Q43P,R35G 8847 K105N,R35V 13717 E87P,Y94V 18587 K101T,S116N 23457 Q43P,R35I 8848 K105N,D36A 13718 E87P,Y94A 18588 K101T,S116K 23458
Q43P,R35L 8849 K105N,D36K 13719 E87P,N95A 18589 K101T,S116H 23459
Q43P,R35M 8850 K105N,R37E 13720 E87P,N95V 18590 K101T,S116Q 23460
Q43P,R35P 8851 K105N,R37A 13721 E87P,T96A 18591 K101T,C117K 23461
Q43P,R35S 8852 K105N,S38A 13722 E87P,K100E 18592 K101T,C117R 23462
Q43P,R35V 8853 K105N,D39A 13723 E87P,K101E 18593 K101T,C117S 23463
Q43P,D36A 8854 K105N,Q40A 13724 E87P,H102A 18594 K101T,C117Q 23464
Q43P,D36K 8855 K105N,Q43A 13725 E87P,K105A 18595 K101T,C117T 23465
Q43P,R37E 8856 K105N,Q43E 13726 E87P,K105E 18596 K101T,C117P 23466
Q43P,R37A 8857 K105N,Q43K 13727 E87P,K113A 18597 K101T,C117A 23467
Q43P,S38A 8858 K105N,S47A 13728 E87P,S116W 18598 K101T,C117I 23468
K101T,K118
Q43P,D39A 8859 K105N,S47V 13729 E87P,S116Y 18599 23469
W
Q43P,Q40A 8860 K105N,A48G 13730 E87P,S116F 18600 K101T,K118H 23470
Q43P,S47A 8861 K105N,E53A 13731 E87P,S116G 18601 K101T,K118R 23471
Q43P,S47V 8862 K105N,Y55A 13732 E87P,S116N 18602 K101T,K118S 23472
K101T,K118
Q43P,A48G 8863 K105N,Y55F 13733 E87P,S116K 18603 23473
M
Q43P,E53A 8864 K105N,K57A 13734 E87P,S116H 18604 K101T,K118L 23474
Q43P,Y55A 8865 K105N,K57E 13735 E87P,S116Q 18605 K101T,P121A 23475
Q43P,Y55F 8866 K105N,E60A 13736 E87P,C117K 18606 K101T,R122A 23476
Q43P,K57A 8867 K105N,E60K 13737 E87P,C117R 18607 K101T,R122E 23477
Q43P,K57E 8868 K105N,T61A 13738 E87P,C117S 18608 K101T,H124A 23478
Q43P,E60A 8869 K105N,Q63A 13739 E87P,C117Q 18609 K101T,Q127A 23479
Q43P,E60K 8870 K105N,Y64A 13740 E87P,C117T 18610 K101T,Q127K 23480
Q43P,T61A 8871 K105N,D68A 13741 E87P,C117P 18611 K101T,K128E 23481
Q43P,Q63A 8872 K105N,T78A 13742 E87P,C117A 18612 K101T,L133A 23482
Q43P,Y64A 8873 K105N,P79A 13743 E87P,C117I 18613 K101T,L133V 23483
Q43P,D68A 8874 K105N,E81K 13744 E87P,K118W 18614 K101V,K10E 23484
Q43P,T78A 8875 K105N,L84A 13745 E87P,K118H 18615 K101V,Y15A 23485
Q43P,P79A 8876 K105N,E87K 13746 E87P,K118R 18616 K101V,Y15F 23486
Q43P,E81K 8877 K105N,E87Q 13747 E87P,K118S 18617 K101V,S17A 23487
Q43P,L84A 8878 K105N,E87V 13748 E87P,K118M 18618 K101V,S17K 23488
Q43P,E87K 8879 K105N,E87D 13749 E87P,K118L 18619 K101V,N18R 23489
Q43P,E87Q 8880 K105N,R88L 13750 E87P,P121A 18620 K101V,H21A 23490
Q43P,E87V 8881 K105N,R88Y 13751 E87P,R122A 18621 K101V,L26A 23491
Q43P,E87D 8882 K105N,R88D 13752 E87P,R122E 18622 K101V,P27A 23492
Q43P,R88L 8883 K105N,L89A 13753 E87P,H124A 18623 K101V,D28K 23493
Q43P,R88Y 8884 K105N,E90K 13754 E87P,Q127A 18624 K101V,T30A 23494
Q43P,R88D 8885 K105N,E91A 13755 E87P,Q127K 18625 K101V,T34A 23495
Q43P,L89A 8886 K105N,N92A 13756 E87P,K128E 18626 K101V,R35A 23496
Q43P,E90K 8887 K105N,Y94V 13757 E87P,L133A 18627 K101V,R35E 23497
Q43P,E91A 8888 K105N,Y94A 13758 E87P,L133V 18628 K101V,R35F 23498
Q43P,N92A 8889 K105N,N95A 13759 E87R,K10E 18629 K101V,R35G 23499
Q43P,Y94V 8890 K105N,N95V 13760 E87R,Y15A 18630 K101V,R35I 23500
Q43P,Y94A 8891 K105N,T96A 13761 E87R,Y15F 18631 K101V,R35L 23501
Q43P,N95A 8892 K105N,K100E 13762 E87R,S17A 18632 K101V,R35M 23502
Figure imgf000227_0001
Figure imgf000228_0001
K101V,H124
Q43S,K57A 8970 K105P,Q63A 13840 E87R,C117P 18710 23580
A
K101V,Q127
Q43S,K57E 8971 K105P,Y64A 13841 E87R,C117A 18711 23581
A
K101V,Q127
Q43S,E60A 8972 K105P,D68A 13842 E87R,C117I 18712 23582
K
E87R,K118
Q43S,E60K 8973 K105P,T78A 13843 18713 K101V,K128E 23583
W
Q43S,T61A 8974 K105P,P79A 13844 E87R,K118H 18714 K101V,L133A 23584
Q43S,Q63A 8975 K105P,E81K 13845 E87R,K118R 18715 K101V,L133V 23585
Q43S,Y64A 8976 K105P,L84A 13846 E87R,K118S 18716 K101W,K10E 23586
E87R,K118
Q43S,D68A 8977 K105P,E87K 13847 18717 K101W,Y15A 23587
M
Q43S,T78A 8978 K105P,E87Q 13848 E87R,K118L 18718 K101W,Y15F 23588
Q43S,P79A 8979 K105P,E87V 13849 E87R,P121A 18719 K101W,S17A 23589
Q43S,E81K 8980 K105P,E87D 13850 E87R,R122A 18720 K101W,S17K 23590
Q43S,L84A 8981 K105P,R88L 13851 E87R,R122E 18721 K101W,N18R 23591
Q43S,E87K 8982 K105P,R88Y 13852 E87R,H124A 18722 K101W,H21A 23592
Q43S,E87Q 8983 K105P,R88D 13853 E87R,Q127A 18723 K101W,L26A 23593
Q43S,E87V 8984 K105P,L89A 13854 E87R,Q127K 18724 K101W,P27A 23594
Q43S,E87D 8985 K105P,E90K 13855 E87R,K128E 18725 K101W,D28K 23595
Q43S,R88L 8986 K105P,E91A 13856 E87R,L133A 18726 K101W,T30A 23596
Q43S,R88Y 8987 K105P,N92A 13857 E87R,L133V 18727 K101W,T34A 23597
Q43S,R88D 8988 K105P,Y94V 13858 E87S,K10E 18728 K101W,R35A 23598
Q43S,L89A 8989 K105P,Y94A 13859 E87S,Y15A 18729 K101W,R35E 23599
Q43S,E90K 8990 K105P,N95A 13860 E87S,Y15F 18730 K101W,R35F 23600
Q43S,E91A 8991 K105P,N95V 13861 E87S,S17A 18731 K101W,R35G 23601
Q43S,N92A 8992 K105P,T96A 13862 E87S,S17K 18732 K101W,R35I 23602
Q43S,Y94V 8993 K105P,K100E 13863 E87S,N18R 18733 K101W,R35L 23603
Q43S,Y94A 8994 K105P,K101E 13864 E87S,H21A 18734 K101W,R35M 23604
Q43S,N95A 8995 K105P,H102A 13865 E87S,L26A 18735 K101W,R35P 23605
Q43S,N95V 8996 K105P,K113A 13866 E87S,P27A 18736 K101W,R35S 23606
Q43S,T96A 8997 K105P,S116W 13867 E87S,D28K 18737 K101W,R35V 23607
Q43S,K100E 8998 K105P,S116Y 13868 E87S,T30A 18738 K101W,D36A 23608
Q43S,K101E 8999 K105P,S116F 13869 E87S,T34A 18739 K101W,D36K 23609
Q43S,H102A 9000 K105P,S116G 13870 E87S,R35A 18740 K101W,R37E 23610
Q43S,K105A 9001 K105P,S116N 13871 E87S,R35E 18741 K101W,R37A 23611
Q43S,K105E 9002 K105P,S116K 13872 E87S,R35F 18742 K101W,S38A 23612
Q43S,K113A 9003 K105P,S116H 13873 E87S,R35G 18743 K101W,D39A 23613
Q43S,S116W 9004 K105P,S116Q 13874 E87S,R35I 18744 K101W,Q40A 23614
Q43S,S116Y 9005 K105P,C117K 13875 E87S,R35L 18745 K101W,Q43A 23615
Q43S,S116F 9006 K105P,C117R 13876 E87S,R35M 18746 K101W,Q43E 23616
Q43S,S116G 9007 K105P,C117S 13877 E87S,R35P 18747 K101W,Q43K 23617
Q43S,S116N 9008 K105P,C117Q 13878 E87S,R35S 18748 K101W,S47A 23618
Q43S,S116K 9009 K105P,C117T 13879 E87S,R35V 18749 K101W,S47V 23619
Q43S,S116H 9010 K105P,C117P 13880 E87S,D36A 18750 K101W,A48G 23620
Q43S,S116Q 9011 K105P,C117A 13881 E87S,D36K 18751 K101W,E53A 23621
Q43S,C117K 9012 K105P,C117I 13882 E87S,R37E 18752 K101W,Y55A 23622 K105P,K118
Q43S,C117R 9013 13883 E87S,R37A 18753 K101W,Y55F 23623
W
Q43S,C117S 9014 K105P,K118H 13884 E87S,S38A 18754 K101W,K57A 23624
Q43S,C117Q 9015 K105P,K118R 13885 E87S,D39A 18755 K101W,K57E 23625
Q43S,C117T 9016 K105P,K118S 13886 E87S,Q40A 18756 K101W,E60A 23626
K105P,K118
Q43S,C117P 9017 13887 E87S,Q43A 18757 K101W,E60K 23627
M
Q43S,C117A 9018 K105P,K118L 13888 E87S,Q43E 18758 K101W,T61A 23628
Q43S,C117I 9019 K105P,P121A 13889 E87S,Q43K 18759 K101W,Q63A 23629
Q43S,K118W 9020 K105P,R122A 13890 E87S,S47A 18760 K101W,Y64A 23630
Q43S,K118H 9021 K105P,R122E 13891 E87S,S47V 18761 K101W,D68A 23631
Q43S,K118R 9022 K105P,H124A 13892 E87S,A48G 18762 K101W,T78A 23632
Q43S,K118S 9023 K105P,Q127A 13893 E87S,E53A 18763 K101W,P79A 23633
Q43S,K118M 9024 K105P,Q127K 13894 E87S,Y55A 18764 K101W,E81K 23634
Q43S,K118L 9025 K105P,K128E 13895 E87S,Y55F 18765 K101W,L84A 23635
Q43S,P121A 9026 K105P,L133A 13896 E87S,K57A 18766 K101W,E87K 23636
Q43S,R122A 9027 K105P,L133V 13897 E87S,K57E 18767 K101W,E87Q 23637
Q43S,R122E 9028 K105Q,K10E 13898 E87S,E60A 18768 K101W,E87V 23638
Q43S,H124A 9029 K105Q,Y15A 13899 E87S,E60K 18769 K101W,E87D 23639
Q43S,Q127A 9030 K105Q,Y15F 13900 E87S,T61A 18770 K101W,R88L 23640
Q43S,Q127K 9031 K105Q,S17A 13901 E87S,Q63A 18771 K101W,R88Y 23641
Q43S,K128E 9032 K105Q,S17K 13902 E87S,Y64A 18772 K101W,R88D 23642
Q43S,L133A 9033 K105Q,N18R 13903 E87S,D68A 18773 K101W,L89A 23643
Q43S,L133V 9034 K105Q,H21A 13904 E87S,T78A 18774 K101W,E90K 23644
Q43T,D28A 9035 K105Q,L26A 13905 E87S,P79A 18775 K101W,E91A 23645
Q43T,E49A 9036 K105Q,P27A 13906 E87S,E81K 18776 K101W,N92A 23646
Q43T,P121A 9037 K105Q,D28K 13907 E87S,L84A 18777 K101W,Y94V 23647
Q43T,K10E 9038 K105Q,T30A 13908 E87S,R88L 18778 K101W,Y94A 23648
Q43T,Y15A 9039 K105Q,T34A 13909 E87S,R88Y 18779 K101W,N95A 23649
Q43T,Y15F 9040 K105Q,R35A 13910 E87S,R88D 18780 K101W,N95V 23650
Q43T,S17A 9041 K105Q,R35E 13911 E87S,L89A 18781 K101W,T96A 23651
K101W,K100
Q43T,S17K 9042 K105Q,R35F 13912 E87S,E90K 18782 23652
E
K101W,H102
Q43T,N18R 9043 K105Q,R35G 13913 E87S,E91A 18783 23653
A
K101W,K105
Q43T,H21A 9044 K105Q,R35I 13914 E87S,N92A 18784 23654
A
K101W,K105
Q43T,L26A 9045 K105Q,R35L 13915 E87S,Y94V 18785 23655
E
K101W,K113
Q43T,P27A 9046 K105Q,R35M 13916 E87S,Y94A 18786 23656
A
K101W,S116
Q43T,D28K 9047 K105Q,R35P 13917 E87S,N95A 18787 23657
W
K101W,S116
Q43T,T30A 9048 K105Q,R35S 13918 E87S,N95V 18788 23658
Y
Q43T,T34A 9049 K105Q,R35V 13919 E87S,T96A 18789 K101W,S116F 23659
K101W,S116
Q43T,R35A 9050 K105Q,D36A 13920 E87S,K100E 18790 23660
G
K101W,S116
Q43T,R35E 9051 K105Q,D36K 13921 E87S,K101E 18791 23661
N
Q43T,R35F 9052 K105Q,R37E 13922 E87S,H102A 18792 K101W,S116 23662
Figure imgf000231_0001
Figure imgf000232_0001
Figure imgf000233_0001
Q43V,D36K 9164 K105R,E53A 14034 E87T,C117R 18904 K101Y,C117I 23774
K101Y,K118
Q43V,R37E 9165 K105R,Y55A 14035 E87T,C117S 18905 23775
W
K101Y,K118
Q43V,R37A 9166 K105R,Y55F 14036 E87T,C117Q 18906 23776
H
K101Y,K118
Q43V,S38A 9167 K105R,K57A 14037 E87T,C117T 18907 23777
R
Q43V,D39A 9168 K105R,K57E 14038 E87T,C117P 18908 K101Y,K118S 23778
K101Y,K118
Q43V,Q40A 9169 K105R,E60A 14039 E87T,C117A 18909 23779
M
Q43V,S47A 9170 K105R,E60K 14040 E87T,C117I 18910 K101Y,K118L 23780
E87T,K118
Q43V,S47V 9171 K105R,T61A 14041 18911 K101Y,P121A 23781
W
K101Y,R122
Q43V,A48G 9172 K105R,Q63A 14042 E87T,K118H 18912 23782
A
Q43V,E53A 9173 K105R,Y64A 14043 E87T,K118R 18913 K101Y,R122E 23783
K101Y,H124
Q43V,Y55A 9174 K105R,D68A 14044 E87T,K118S 18914 23784
A
K101Y,Q127
Q43V,Y55F 9175 K105R,T78A 14045 E87T,K118M 18915 23785
A
K101Y,Q127
Q43V,K57A 9176 K105R,P79A 14046 E87T,K118L 18916 23786
K
Q43V,K57E 9177 K105R,E81K 14047 E87T,P121A 18917 K101Y,K128E 23787
Q43V,E60A 9178 K105R,L84A 14048 E87T,R122A 18918 K101Y,L133A 23788
Q43V,E60K 9179 K105R,E87K 14049 E87T,R122E 18919 K101Y,L133V 23789
Q43V,T61A 9180 K105R,E87Q 14050 E87T,H124A 18920 C117D,K10E 23790
Q43V,Q63A 9181 K105R,E87V 14051 E87T,Q127A 18921 C117D,Y15A 23791
Q43V,Y64A 9182 K105R,E87D 14052 E87T,Q127K 18922 C117D,Y15F 23792
Q43V,D68A 9183 K105R,R88L 14053 E87T,K128E 18923 C117D,S17A 23793
Q43V,T78A 9184 K105R,R88Y 14054 E87T,L133A 18924 C117D,S17K 23794
Q43V,P79A 9185 K105R,R88D 14055 E87T,L133V 18925 C117D,N18R 23795
Q43V,E81K 9186 K105R,L89A 14056 E87W,K10E 18926 C117D,H21A 23796
Q43V,L84A 9187 K105R,E90K 14057 E87W,Y15A 18927 C117D,L26A 23797
Q43V,E87K 9188 K105R,E91A 14058 E87W,Y15F 18928 C117D,P27A 23798
Q43V,E87Q 9189 K105R,N92A 14059 E87W,S17A 18929 C117D,D28K 23799
Q43V,E87V 9190 K105R,Y94V 14060 E87W,S17K 18930 C117D,T30A 23800
Q43V,E87D 9191 K105R,Y94A 14061 E87W,N18R 18931 C117D,T34A 23801
Q43V,R88L 9192 K105R,N95A 14062 E87W,H21A 18932 C117D,R35A 23802
Q43V,R88Y 9193 K105R,N95V 14063 E87W,L26A 18933 C117D,R35E 23803
Q43V,R88D 9194 K105R,T96A 14064 E87W,P27A 18934 C117D,R35F 23804
Q43V,L89A 9195 K105R,K100E 14065 E87W,D28K 18935 C117D,R35G 23805
Q43V,E90K 9196 K105R,K101E 14066 E87W,T30A 18936 C117D,R35I 23806
K105R,H102
Q43V,E91A 9197 14067 E87W,T34A 18937 C117D,R35L 23807
A
K105R,K113
Q43V,N92A 9198 14068 E87W,R35A 18938 C117D,R35M 23808
A
K105R,S116
Q43V,Y94V 9199 14069 E87W,R35E 18939 C117D,R35P 23809
W
Q43V,Y94A 9200 K105R,S116Y 14070 E87W,R35F 18940 C117D,R35S 23810
Q43V,N95A 9201 K105R,S116F 14071 E87W,R35G 18941 C117D,R35V 23811
Q43V,N95V 9202 K105R,S116G 14072 E87W,R35I 18942 C117D,D36A 23812 Q43V,T96A 9203 K105R,S116N 14073 E87W,R35L 18943 C117D,D36K 23813
Q43V,K100E 9204 K105R,S116K 14074 E87W,R35M 18944 C117D,R37E 23814
Q43V,K101E 9205 K105R,S116H 14075 E87W,R35P 18945 C117D,R37A 23815
Q43V,H102A 9206 K105R,S116Q 14076 E87W,R35S 18946 C117D,S38A 23816
Q43V,K105A 9207 K105R,C117K 14077 E87W,R35V 18947 C117D,D39A 23817
Q43V,K105E 9208 K105R,C117R 14078 E87W,D36A 18948 C117D,Q40A 23818
Q43V,K113A 9209 K105R,C117S 14079 E87W,D36K 18949 C117D,Q43A 23819
Q43V,S116W 9210 K105R,C117Q 14080 E87W,R37E 18950 C117D,Q43E 23820
Q43V,S116Y 9211 K105R,C117T 14081 E87W,R37A 18951 C117D,Q43K 23821
Q43V,S116F 9212 K105R,C117P 14082 E87W,S38A 18952 C117D,S47A 23822
Q43V,S116G 9213 K105R,C117A 14083 E87W,D39A 18953 C117D,S47V 23823
Q43V,S116N 9214 K105R,C117I 14084 E87W,Q40A 18954 C117D,A48G 23824
K105R,K118
Q43V,S116K 9215 14085 E87W,Q43A 18955 C117D,E53A 23825
W
K105R,K118
Q43V,S116H 9216 14086 E87W,Q43E 18956 C117D,Y55A 23826
H
Q43V,S116Q 9217 K105R,K118R 14087 E87W,Q43K 18957 C117D,Y55F 23827
Q43V,C117K 9218 K105R,K118S 14088 E87W,S47A 18958 C117D,K57A 23828
K105R,K118
Q43V,C117R 9219 14089 E87W,S47V 18959 C117D,K57E 23829
M
Q43V,C117S 9220 K105R,K118L 14090 E87W,A48G 18960 C117D,E60A 23830
Q43V,C117Q 9221 K105R,P121A 14091 E87W,E53A 18961 C117D,E60K 23831
Q43V,C117T 9222 K105R,R122A 14092 E87W,Y55A 18962 C117D,T61A 23832
Q43V,C117P 9223 K105R,R122E 14093 E87W,Y55F 18963 C117D,Q63A 23833
K105R,H124
Q43V,C117A 9224 14094 E87W,K57A 18964 C117D,Y64A 23834
A
K105R,Q127
Q43V,C117I 9225 14095 E87W,K57E 18965 C117D,D68A 23835
A
Q43V,K118 K105R,Q127
9226 14096 E87W,E60A 18966 C117D,T78A 23836 W K
Q43V,K118H 9227 K105R,K128E 14097 E87W,E60K 18967 C117D,P79A 23837
Q43V,K118R 9228 K105R,L133A 14098 E87W,T61A 18968 C117D,E81K 23838
Q43V,K118S 9229 K105R,L133V 14099 E87W,Q63A 18969 C117D,L84A 23839
Q43V,K118
9230 K105S,K10E 14100 E87W,Y64A 18970 C117D,E87K 23840 M
Q43V,K118L 9231 K105S,Y15A 14101 E87W,D68A 18971 C117D,E87Q 23841
Q43V,P121A 9232 K105S,Y15F 14102 E87W,T78A 18972 C117D,E87V 23842
Q43V,R122A 9233 K105S,S17A 14103 E87W,P79A 18973 C117D,E87D 23843
Q43V,R122E 9234 K105S,S17K 14104 E87W,E81K 18974 C117D,R88L 23844
Q43V,H124A 9235 K105S,N18R 14105 E87W,L84A 18975 C117D,R88Y 23845
Q43V,Q127A 9236 K105S,H21A 14106 E87W,R88L 18976 C117D,R88D 23846
Q43V,Q127K 9237 K105S,L26A 14107 E87W,R88Y 18977 C117D,L89A 23847
Q43V,K128E 9238 K105S,P27A 14108 E87W,R88D 18978 C117D,E90K 23848
Q43V,L133A 9239 K105S,D28K 14109 E87W,L89A 18979 C117D,E91A 23849
Q43V,L133V 9240 K105S,T30A 14110 E87W,E90K 18980 C117D,N92A 23850
Q43W,D28A 9241 K105S,T34A 14111 E87W,E91A 18981 C117D,Y94V 23851
Q43W,E49A 9242 K105S,R35A 14112 E87W,N92A 18982 C117D,Y94A 23852
Q43W,P121A 9243 K105S,R35E 14113 E87W,Y94V 18983 C117D,N95A 23853
Q43W,K10E 9244 K105S,R35F 14114 E87W,Y94A 18984 C117D,N95V 23854
Figure imgf000236_0001
E87W,H124
Q43W,K57A 9279 K105S,L84A 14149 19019 C117E,S17K 23889
A
E87W,Q127
Q43W,K57E 9280 K105S,E87K 14150 19020 C117E,N18R 23890
A
E87W,Q127
Q43W,E60A 9281 K105S,E87Q 14151 19021 C117E,H21A 23891
K
E87W,K128
Q43W,E60K 9282 K105S,E87V 14152 19022 C117E,L26A 23892
E
E87W,L133
Q43W,T61A 9283 K105S,E87D 14153 19023 C117E,P27A 23893
A
E87W,L133
Q43W,Q63A 9284 K105S,R88L 14154 19024 C117E,D28K 23894
V
Q43W,Y64A 9285 K105S,R88Y 14155 E87Y,K10E 19025 C117E,T30A 23895
Q43W,D68A 9286 K105S,R88D 14156 E87Y,Y15A 19026 C117E,T34A 23896
Q43W,T78A 9287 K105S,L89A 14157 E87Y,Y15F 19027 C117E,R35A 23897
Q43W,P79A 9288 K105S,E90K 14158 E87Y,S17A 19028 C117E,R35E 23898
Q43W,E81K 9289 K105S,E91A 14159 E87Y,S17K 19029 C117E,R35F 23899
Q43W,L84A 9290 K105S,N92A 14160 E87Y,N18R 19030 C117E,R35G 23900
Q43W,E87K 9291 K105S,Y94V 14161 E87Y,H21A 19031 C117E,R35I 23901
Q43W,E87Q 9292 K105S,Y94A 14162 E87Y,L26A 19032 C117E,R35L 23902
Q43W,E87V 9293 K105S,N95A 14163 E87Y,P27A 19033 C117E,R35M 23903
Q43W,E87D 9294 K105S,N95V 14164 E87Y,D28K 19034 C117E,R35P 23904
Q43W,R88L 9295 K105S,T96A 14165 E87Y,T30A 19035 C117E,R35S 23905
Q43W,R88Y 9296 K105S,K100E 14166 E87Y,T34A 19036 C117E,R35V 23906
Q43W,R88D 9297 K105S,K101E 14167 E87Y,R35A 19037 C117E,D36A 23907
Q43W,L89A 9298 K105S,H102A 14168 E87Y,R35E 19038 C117E,D36K 23908
Q43W,E90K 9299 K105S,K113A 14169 E87Y,R35F 19039 C117E,R37E 23909
Q43W,E91A 9300 K105S,S116W 14170 E87Y,R35G 19040 C117E,R37A 23910
Q43W,N92A 9301 K105S,S116Y 14171 E87Y,R35I 19041 C117E,S38A 23911
Q43W,Y94V 9302 K105S,S116F 14172 E87Y,R35L 19042 C117E,D39A 23912
Q43W,Y94A 9303 K105S,S116G 14173 E87Y,R35M 19043 C117E,Q40A 23913
Q43W,N95A 9304 K105S,S116N 14174 E87Y,R35P 19044 C117E,Q43A 23914
Q43W,N95V 9305 K105S,S116K 14175 E87Y,R35S 19045 C117E,Q43E 23915
Q43W,T96A 9306 K105S,S116H 14176 E87Y,R35V 19046 C117E,Q43K 23916
Q43W,K100
9307 K105S,S116Q 14177 E87Y,D36A 19047 C117E,S47A 23917 E
Q43W,K101
9308 K105S,C117K 14178 E87Y,D36K 19048 C117E,S47V 23918 E
Q43W,H102
9309 K105S,C117R 14179 E87Y,R37E 19049 C117E,A48G 23919 A
Q43W,K105
9310 K105S,C117S 14180 E87Y,R37A 19050 C117E,E53A 23920 A
Q43W,K105
9311 K105S,C117Q 14181 E87Y,S38A 19051 C117E,Y55A 23921 E
Q43W,K113
9312 K105S,C117T 14182 E87Y,D39A 19052 C117E,Y55F 23922 A
Q43W,S116
9313 K105S,C117P 14183 E87Y,Q40A 19053 C117E,K57A 23923 W
Q43W,S116Y 9314 K105S,C117A 14184 E87Y,Q43A 19054 C117E,K57E 23924
Q43W,S116F 9315 K105S,C117I 14185 E87Y,Q43E 19055 C117E,E60A 23925
K105S,K118
Q43W,S116G 9316 14186 E87Y,Q43K 19056 C117E,E60K 23926
W
Figure imgf000238_0001
Q43Y,H21A 9353 K105T,D36A 14223 E87Y,S116W 19093 C117E,S116H 23963
Q43Y,L26A 9354 K105T,D36K 14224 E87Y,S116Y 19094 C117E,S116Q 23964
C117E,K118
Q43Y,P27A 9355 K105T,R37E 14225 E87Y,S116F 19095 23965
W
Q43Y,D28K 9356 K105T,R37A 14226 E87Y,S116G 19096 C117E,K118H 23966
Q43Y,T30A 9357 K105T,S38A 14227 E87Y,S116N 19097 C117E,K118R 23967
Q43Y,T34A 9358 K105T,D39A 14228 E87Y,S116K 19098 C117E,K118S 23968
C117E,K118
Q43Y,R35A 9359 K105T,Q40A 14229 E87Y,S116H 19099 23969
M
Q43Y,R35E 9360 K105T,Q43A 14230 E87Y,S116Q 19100 C117E,K118L 23970
Q43Y,R35F 9361 K105T,Q43E 14231 E87Y,C117K 19101 C117E,P121A 23971
Q43Y,R35G 9362 K105T,Q43K 14232 E87Y,C117R 19102 C117E,R122A 23972
Q43Y,R35I 9363 K105T,S47A 14233 E87Y,C117S 19103 CI 17E,R122E 23973
Q43Y,R35L 9364 K105T,S47V 14234 E87Y,C117Q 19104 C117E,H124A 23974
Q43Y,R35M 9365 K105T,A48G 14235 E87Y,C117T 19105 C117E,Q127A 23975
Q43Y,R35P 9366 K105T,E53A 14236 E87Y,C117P 19106 C117E,Q127K 23976
Q43Y,R35S 9367 K105T,Y55A 14237 E87Y,C117A 19107 C117E,K128E 23977
Q43Y,R35V 9368 K105T,Y55F 14238 E87Y,C117I 19108 C117E,L133A 23978
E87Y,K118
Q43Y,D36A 9369 K105T,K57A 14239 19109 C117E,L133V 23979
W
Q43Y,D36K 9370 K105T,K57E 14240 E87Y,K118H 19110 C117F,K10E 23980
Q43Y,R37E 9371 K105T,E60A 14241 E87Y,K118R 19111 C117F,Y15A 23981
Q43Y,R37A 9372 K105T,E60K 14242 E87Y,K118S 19112 C117F,Y15F 23982
E87Y,K118
Q43Y,S38A 9373 K105T,T61A 14243 19113 C117F,S17A 23983
M
Q43Y,D39A 9374 K105T,Q63A 14244 E87Y,K118L 19114 C117F,S17K 23984
Q43Y,Q40A 9375 K105T,Y64A 14245 E87Y,P121A 19115 C117F,N18R 23985
Q43Y,S47A 9376 K105T,D68A 14246 E87Y,R122A 19116 C117F,H21A 23986
Q43Y,S47V 9377 K105T,T78A 14247 E87Y,R122E 19117 C117F,L26A 23987
Q43Y,A48G 9378 K105T,P79A 14248 E87Y,H124A 19118 C117F,P27A 23988
Q43Y,E53A 9379 K105T,E81K 14249 E87Y,Q127A 19119 C117F,D28K 23989
Q43Y,Y55A 9380 K105T,L84A 14250 E87Y,Q127K 19120 C117F,T30A 23990
Q43Y,Y55F 9381 K105T,E87K 14251 E87Y,K128E 19121 C117F,T34A 23991
Q43Y,K57A 9382 K105T,E87Q 14252 E87Y,L133A 19122 C117F,R35A 23992
Q43Y,K57E 9383 K105T,E87V 14253 E87Y,L133V 19123 C117F,R35E 23993
Q43Y,E60A 9384 K105T,E87D 14254 R88F,K10E 19124 C117F,R35F 23994
Q43Y,E60K 9385 K105T,R88L 14255 R88F,Y15A 19125 C117F,R35G 23995
Q43Y,T61A 9386 K105T,R88Y 14256 R88F,Y15F 19126 C117F,R35I 23996
Q43Y,Q63A 9387 K105T,R88D 14257 R88F,S17A 19127 C117F,R35L 23997
Q43Y,Y64A 9388 K105T,L89A 14258 R88F,S17K 19128 C117F,R35M 23998
Q43Y,D68A 9389 K105T,E90K 14259 R88F,N18R 19129 C117F,R35P 23999
Q43Y,T78A 9390 K105T,E91A 14260 R88F,H21A 19130 C117F,R35S 24000
Q43Y,P79A 9391 K105T,N92A 14261 R88F,L26A 19131 C117F,R35V 24001
Q43Y,E81K 9392 K105T,Y94V 14262 R88F,P27A 19132 C117F,D36A 24002
Q43Y,L84A 9393 K105T,Y94A 14263 R88F,D28K 19133 C117F,D36K 24003
Q43Y,E87K 9394 K105T,N95A 14264 R88F,T30A 19134 C117F,R37E 24004
Q43Y,E87Q 9395 K105T,N95V 14265 R88F,T34A 19135 C117F,R37A 24005
Q43Y,E87V 9396 K105T,T96A 14266 R88F,R35A 19136 C117F,S38A 24006 Q43Y,E87D 9397 K105T,K100E 14267 R88F,R35E 19137 C117F,D39A 24007
Q43Y,R88L 9398 K105T,K101E 14268 R88F,R35F 19138 C117F,Q40A 24008
Q43Y,R88Y 9399 K105T,H102A 14269 R88F,R35G 19139 C117F,Q43A 24009
Q43Y,R88D 9400 K105T,K113A 14270 R88F,R35I 19140 C117F,Q43E 24010
K105T,S116
Q43Y,L89A 9401 14271 R88F,R35L 19141 C117F,Q43K 24011
W
Q43Y,E90K 9402 K105T,S116Y 14272 R88F,R35M 19142 C117F,S47A 24012
Q43Y,E91A 9403 K105T,S116F 14273 R88F,R35P 19143 C117F,S47V 24013
Q43Y,N92A 9404 K105T,S116G 14274 R88F,R35S 19144 C117F,A48G 24014
Q43Y,Y94V 9405 K105T,S116N 14275 R88F,R35V 19145 C117F,E53A 24015
Q43Y,Y94A 9406 K105T,S116K 14276 R88F,D36A 19146 C117F,Y55A 24016
Q43Y,N95A 9407 K105T,S116H 14277 R88F,D36K 19147 C117F,Y55F 24017
Q43Y,N95V 9408 K105T,S116Q 14278 R88F,R37E 19148 C117F,K57A 24018
Q43Y,T96A 9409 K105T,C117K 14279 R88F,R37A 19149 C117F,K57E 24019
Q43Y,K100E 9410 K105T,C117R 14280 R88F,S38A 19150 C117F,E60A 24020
Q43Y,K101E 9411 K105T,C117S 14281 R88F,D39A 19151 C117F,E60K 24021
Q43Y,H102A 9412 K105T,C117Q 14282 R88F,Q40A 19152 C117F,T61A 24022
Q43Y,K105A 9413 K105T,C117T 14283 R88F,Q43A 19153 C117F,Q63A 24023
Q43Y,K105E 9414 K105T,C117P 14284 R88F,Q43E 19154 C117F,Y64A 24024
Q43Y,K113A 9415 K105T,C117A 14285 R88F,Q43K 19155 C117F,D68A 24025
Q43Y,S116W 9416 K105T,C117I 14286 R88F,S47A 19156 C117F,T78A 24026
K105T,K118
Q43Y,S116Y 9417 14287 R88F,S47V 19157 C117F,P79A 24027
W
Q43Y,S116F 9418 K105T,K118H 14288 R88F,A48G 19158 C117F,E81K 24028
Q43Y,S116G 9419 K105T,K118R 14289 R88F,E53A 19159 C117F,L84A 24029
Q43Y,S116N 9420 K105T,K118S 14290 R88F,Y55A 19160 C117F,E87K 24030
K105T,K118
Q43Y,S116K 9421 14291 R88F,Y55F 19161 C117F,E87Q 24031
M
Q43Y,S116H 9422 K105T,K118L 14292 R88F,K57A 19162 C117F,E87V 24032
Q43Y,S116Q 9423 K105T,P121A 14293 R88F,K57E 19163 C117F,E87D 24033
Q43Y,C117K 9424 K105T,R122A 14294 R88F,E60A 19164 C117F,R88L 24034
Q43Y,C117R 9425 K105T,R122E 14295 R88F,E60K 19165 C117F,R88Y 24035
Q43Y,C117S 9426 K105T,H124A 14296 R88F,T61A 19166 C117F,R88D 24036
Q43Y,C117Q 9427 K105T,Q127A 14297 R88F,Q63A 19167 C117F,L89A 24037
Q43Y,C117T 9428 K105T,Q127K 14298 R88F,Y64A 19168 C117F,E90K 24038
Q43Y,C117P 9429 K105T,K128E 14299 R88F,D68A 19169 C117F,E91A 24039
Q43Y,C117A 9430 K105T,L133A 14300 R88F,T78A 19170 C117F,N92A 24040
Q43Y,C117I 9431 K105T,L133V 14301 R88F,P79A 19171 C117F,Y94V 24041
Q43Y,K118
9432 K105V,K10E 14302 R88F,E81K 19172 C117F,Y94A 24042 W
Q43Y,K118H 9433 K105V,Y15A 14303 R88F,L84A 19173 C117F,N95A 24043
Q43Y,K118R 9434 K105V,Y15F 14304 R88F,E87K 19174 C117F,N95V 24044
Q43Y,K118S 9435 K105V,S17A 14305 R88F,E87Q 19175 C117F,T96A 24045
Q43Y,K118
9436 K105V,S17K 14306 R88F,E87V 19176 C117F,K100E 24046 M
Q43Y,K118L 9437 K105V,N18R 14307 R88F,E87D 19177 C117F,K101E 24047
Q43Y,P121A 9438 K105V,H21A 14308 R88F,L89A 19178 C117F,H102A 24048
Q43Y,R122A 9439 K105V,L26A 14309 R88F,E90K 19179 C117F,K105A 24049 Q43Y,R122E 9440 K105V,P27A 14310 R88F,E91A 19180 C117F,K105E 24050
Q43Y,H124A 9441 K105V,D28K 14311 R88F,N92A 19181 C117F,K113A 24051
Q43Y,Q127A 9442 K105V,T30A 14312 R88F,Y94V 19182 C117F,S116W 24052
Q43Y,Q127K 9443 K105V,T34A 14313 R88F,Y94A 19183 C117F,S116Y 24053
Q43Y,K128E 9444 K105V,R35A 14314 R88F,N95A 19184 C117F,S116F 24054
Q43Y,L133A 9445 K105V,R35E 14315 R88F,N95V 19185 C117F,S116G 24055
Q43Y,L133V 9446 K105V,R35F 14316 R88F,T96A 19186 C117F,S116N 24056
D28A,E49A 9447 K105V,R35G 14317 R88F,K100E 19187 C117F,S116K 24057
D28A,P121A 9448 K105V,R35I 14318 R88F,K101E 19188 C117F,S116H 24058
D28A,K10E 9449 K105V,R35L 14319 R88F,H102A 19189 C117F,S116Q 24059
C117F,K118
D28A,Y15A 9450 K105V,R35M 14320 R88F,K105A 19190 24060
W
D28A,Y15F 9451 K105V,R35P 14321 R88F,K105E 19191 C117F,K118H 24061
D28A,S17A 9452 K105V,R35S 14322 R88F,K113A 19192 C117F,K118R 24062
D28A,S17K 9453 K105V,R35V 14323 R88F,S116W 19193 C117F,K118S 24063
C117F,K118
D28A,N18R 9454 K105V,D36A 14324 R88F,S116Y 19194 24064
M
D28A,H21A 9455 K105V,D36K 14325 R88F,S116F 19195 C117F,K118L 24065
D28A,L26A 9456 K105V,R37E 14326 R88F,S116G 19196 C117F,P121A 24066
D28A,P27A 9457 K105V,R37A 14327 R88F,S116N 19197 C117F,R122A 24067
D28A,T30A 9458 K105V,S38A 14328 R88F,S116K 19198 C117F,R122E 24068
D28A,T34A 9459 K105V,D39A 14329 R88F,S116H 19199 C117F,H124A 24069
D28A,R35A 9460 K105V,Q40A 14330 R88F,S116Q 19200 C117F,Q127A 24070
D28A,R35E 9461 K105V,Q43A 14331 R88F,C117K 19201 C117F,Q127K 24071
D28A,R35F 9462 K105V,Q43E 14332 R88F,C117R 19202 C117F,K128E 24072
D28A,R35G 9463 K105V,Q43K 14333 R88F,C117S 19203 C117F,L133A 24073
D28A,R35I 9464 K105V,S47A 14334 R88F,C117Q 19204 C117F,L133V 24074
D28A,R35L 9465 K105V,S47V 14335 R88F,C117T 19205 C117G,K10E 24075
D28A,R35M 9466 K105V,A48G 14336 R88F,C117P 19206 C117G,Y15A 24076
D28A,R35P 9467 K105V,E53A 14337 R88F,C117A 19207 C117G,Y15F 24077
D28A,R35S 9468 K105V,Y55A 14338 R88F,C117I 19208 C117G,S17A 24078
R88F,K118
D28A,R35V 9469 K105V,Y55F 14339 19209 C117G,S17K 24079
W
D28A,D36A 9470 K105V,K57A 14340 R88F,K118H 19210 C117G,N18R 24080
D28A,D36K 9471 K105V,K57E 14341 R88F,K118R 19211 C117G,H21A 24081
D28A,R37E 9472 K105V,E60A 14342 R88F,K118S 19212 C117G,L26A 24082
D28A,R37A 9473 K105V,E60K 14343 R88F,K118M 19213 C117G,P27A 24083
D28A,S38A 9474 K105V,T61A 14344 R88F,K118L 19214 C117G,D28K 24084
D28A,D39A 9475 K105V,Q63A 14345 R88F,P121A 19215 C117G,T30A 24085
D28A,Q40A 9476 K105V,Y64A 14346 R88F,R122A 19216 C117G,T34A 24086
D28A,Q43A 9477 K105V,D68A 14347 R88F,R122E 19217 C117G,R35A 24087
D28A,Q43E 9478 K105V,T78A 14348 R88F,H124A 19218 C117G,R35E 24088
D28A,Q43K 9479 K105V,P79A 14349 R88F,Q127A 19219 C117G,R35F 24089
D28A,S47A 9480 K105V,E81K 14350 R88F,Q127K 19220 C117G,R35G 24090
D28A,S47V 9481 K105V,L84A 14351 R88F,K128E 19221 C117G,R35I 24091
D28A,A48G 9482 K105V,E87K 14352 R88F,L133A 19222 C117G,R35L 24092
D28A,E53A 9483 K105V,E87Q 14353 R88F,L133V 19223 C117G,R35M 24093
D28A,Y55A 9484 K105V,E87V 14354 R88G,K10E 19224 C117G,R35P 24094
Figure imgf000242_0001
A
D28A,S116H 9526 K105V,R122E 14396 R88G,T61A 19266 C117G,Y94V 24136
K105V,H124
D28A,S116Q 9527 14397 R88G,Q63A 19267 C117G,Y94A 24137
A
K105V,Q127
D28A,C117K 9528 14398 R88G,Y64A 19268 C117G,N95A 24138
A
K105V,Q127
D28A,C117R 9529 14399 R88G,D68A 19269 C117G,N95V 24139
K
D28A,C117S 9530 K105V,K128E 14400 R88G,T78A 19270 C117G,T96A 24140
D28A,C117Q 9531 K105V,L133A 14401 R88G,P79A 19271 C117G,K100E 24141
D28A,C117T 9532 K105V,L133V 14402 R88G,E81K 19272 C117G,K101E 24142
C117G,H102
D28A,C117P 9533 K105W,K10E 14403 R88G,L84A 19273 24143
A
C117G,K105
D28A,C117A 9534 K105W,Y15A 14404 R88G,E87K 19274 24144
A
D28A,C117I 9535 K105W,Y15F 14405 R88G,E87Q 19275 C117G,K105E 24145
D28A,K118 C117G,K113
9536 K105W,S17A 14406 R88G,E87V 19276 24146 W A
C117G,S116
D28A,K118H 9537 K105W,S17K 14407 R88G,E87D 19277 24147
W
D28A,K118R 9538 K105W,N18R 14408 R88G,L89A 19278 C117G,S116Y 24148
D28A,K118S 9539 K105W,H21A 14409 R88G,E90K 19279 C117G,S116F 24149
D28A,K118
9540 K105W,L26A 14410 R88G,E91A 19280 C117G,S116G 24150 M
D28A,K118L 9541 K105W,P27A 14411 R88G,N92A 19281 C117G,S116N 24151
D28A,P121A 9542 K105W,D28K 14412 R88G,Y94V 19282 C117G,S116K 24152
D28A,R122A 9543 K105W,T30A 14413 R88G,Y94A 19283 C117G,S116H 24153
D28A,R122E 9544 K105W,T34A 14414 R88G,N95A 19284 C117G,S116Q 24154
C117G,K118
D28A,H124A 9545 K105W,R35A 14415 R88G,N95V 19285 24155
W
C117G,K118
D28A,Q127A 9546 K105W,R35E 14416 R88G,T96A 19286 24156
H
D28A,Q127K 9547 K105W,R35F 14417 R88G,K100E 19287 C117G,K118R 24157
D28A,K128E 9548 K105W,R35G 14418 R88G,K101E 19288 C117G,K118S 24158
C117G,K118
D28A,L133A 9549 K105W,R35I 14419 R88G,H102A 19289 24159
M
D28A,L133V 9550 K105W,R35L 14420 R88G,K105A 19290 C117G,K118L 24160
E49A,P121A 9551 K105W,R35M 14421 R88G,K105E 19291 C117G,P121A 24161
E49A,K10E 9552 K105W,R35P 14422 R88G,K113A 19292 C117G,R122A 24162
R88G,S116
E49A,Y15A 9553 K105W,R35S 14423 19293 C117G,R122E 24163
W
C117G,H124
E49A,Y15F 9554 K105W,R35V 14424 R88G,S116Y 19294 24164
A
C117G,Q127
E49A,S17A 9555 K105W,D36A 14425 R88G,S116F 19295 24165
A
C117G,Q127
E49A,S17K 9556 K105W,D36K 14426 R88G,S116G 19296 24166
K
E49A,N18R 9557 K105W,R37E 14427 R88G,S116N 19297 C117G,K128E 24167
E49A,H21A 9558 K105W,R37A 14428 R88G,S116K 19298 C117G,L133A 24168
E49A,L26A 9559 K105W,S38A 14429 R88G,S116H 19299 C117G,L133V 24169
E49A,P27A 9560 K105W,D39A 14430 R88G,S116Q 19300 C117H,K10E 24170
E49A,D28K 9561 K105W,Q40A 14431 R88G,C117K 19301 C117H,Y15A 24171
E49A,T30A 9562 K105W,Q43A 14432 R88G,C117R 19302 C117H,Y15F 24172 E49A,T34A 9563 K105W,Q43E 14433 R88G,C117S 19303 C117H,S17A 24173
E49A,R35A 9564 K105W,Q43K 14434 R88G,C117Q 19304 C117H,S17K 24174
E49A,R35E 9565 K105W,S47A 14435 R88G,C117T 19305 C117H,N18R 24175
E49A,R35F 9566 K105W,S47V 14436 R88G,C117P 19306 C117H,H21A 24176
E49A,R35G 9567 K105W,A48G 14437 R88G,C117A 19307 C117H,L26A 24177
E49A,R35I 9568 K105W,E53A 14438 R88G,C117I 19308 C117H,P27A 24178
R88G,K118
E49A,R35L 9569 K105W,Y55A 14439 19309 C117H,D28K 24179
W
E49A,R35M 9570 K105W,Y55F 14440 R88G,K118H 19310 C117H,T30A 24180
E49A,R35P 9571 K105W,K57A 14441 R88G,K118R 19311 C117H,T34A 24181
E49A,R35S 9572 K105W,K57E 14442 R88G,K118S 19312 C117H,R35A 24182
R88G,K118
E49A,R35V 9573 K105W,E60A 14443 19313 C117H,R35E 24183
M
E49A,D36A 9574 K105W,E60K 14444 R88G,K118L 19314 C117H,R35F 24184
E49A,D36K 9575 K105W,T61A 14445 R88G,P121A 19315 C117H,R35G 24185
E49A,R37E 9576 K105W,Q63A 14446 R88G,R122A 19316 C117H,R35I 24186
E49A,R37A 9577 K105W,Y64A 14447 R88G,R122E 19317 C117H,R35L 24187
E49A,S38A 9578 K105W,D68A 14448 R88G,H124A 19318 C117H,R35M 24188
E49A,D39A 9579 K105W,T78A 14449 R88G,Q127A 19319 C117H,R35P 24189
E49A,Q40A 9580 K105W,P79A 14450 R88G,Q127K 19320 C117H,R35S 24190
E49A,Q43A 9581 K105W,E81K 14451 R88G,K128E 19321 C117H,R35V 24191
E49A,Q43E 9582 K105W,L84A 14452 R88G,L133A 19322 C117H,D36A 24192
E49A,Q43K 9583 K105W,E87K 14453 R88G,L133V 19323 C117H,D36K 24193
E49A,S47A 9584 K105W,E87Q 14454 R88H,K10E 19324 C117H,R37E 24194
E49A,S47V 9585 K105W,E87V 14455 R88H,Y15A 19325 C117H,R37A 24195
E49A,A48G 9586 K105W,E87D 14456 R88H,Y15F 19326 C117H,S38A 24196
E49A,E53A 9587 K105W,R88L 14457 R88H,S17A 19327 C117H,D39A 24197
E49A,Y55A 9588 K105W,R88Y 14458 R88H,S17K 19328 C117H,Q40A 24198
E49A,Y55F 9589 K105W,R88D 14459 R88H,N18R 19329 C117H,Q43A 24199
E49A,K57A 9590 K105W,L89A 14460 R88H,H21A 19330 C117H,Q43E 24200
E49A,K57E 9591 K105W,E90K 14461 R88H,L26A 19331 C117H,Q43K 24201
E49A,E60A 9592 K105W,E91A 14462 R88H,P27A 19332 C117H,S47A 24202
E49A,E60K 9593 K105W,N92A 14463 R88H,D28K 19333 C117H,S47V 24203
E49A,T61A 9594 K105W,Y94V 14464 R88H,T30A 19334 C117H,A48G 24204
E49A,Q63A 9595 K105W,Y94A 14465 R88H,T34A 19335 C117H,E53A 24205
E49A,Y64A 9596 K105W,N95A 14466 R88H,R35A 19336 C117H,Y55A 24206
E49A,D68A 9597 K105W,N95V 14467 R88H,R35E 19337 C117H,Y55F 24207
E49A,T78A 9598 K105W,T96A 14468 R88H,R35F 19338 C117H,K57A 24208
K105W,K100
E49A,P79A 9599 14469 R88H,R35G 19339 C117H,K57E 24209
E
K105W,K101
E49A,E81K 9600 14470 R88H,R35I 19340 C117H,E60A 24210
E
K105W,H102
E49A,L84A 9601 14471 R88H,R35L 19341 C117H,E60K 24211
A
K105W,K113
E49A,E87K 9602 14472 R88H,R35M 19342 C117H,T61A 24212
A
K105W,S116
E49A,E87Q 9603 14473 R88H,R35P 19343 C117H,Q63A 24213
W
K105W,S116
E49A,E87V 9604 14474 R88H,R35S 19344 C117H,Y64A 24214
Y
Figure imgf000245_0001
E49A,C117T 9636 K105Y,Y15F 14506 R88H,E87V 19376 C117H,S116N 24246
E49A,C117P 9637 K105Y,S17A 14507 R88H,E87D 19377 C117H,S116K 24247
E49A,C117A 9638 K105Y,S17K 14508 R88H,L89A 19378 C117H,S116H 24248
E49A,C117I 9639 K105Y,N18R 14509 R88H,E90K 19379 C117H,S116Q 24249
E49A,K118 C117H,K118
9640 K105Y,H21A 14510 R88H,E91A 19380 24250 W W
C117H,K118
E49A,K118H 9641 K105Y,L26A 14511 R88H,N92A 19381 24251
H
E49A,K118R 9642 K105Y,P27A 14512 R88H,Y94V 19382 C117H,K118R 24252
E49A,K118S 9643 K105Y,D28K 14513 R88H,Y94A 19383 C117H,K118S 24253
C117H,K118
E49A,K118M 9644 Κ105Υ/Γ30Α 14514 R88H,N95A 19384 24254
M
E49A,K118L 9645 Κ105Υ/Γ34Α 14515 R88H,N95V 19385 C117H,K118L 24255
E49A,P121A 9646 K105Y,R35A 14516 R88H,T96A 19386 C117H,P121A 24256
E49A,R122A 9647 K105Y,R35E 14517 R88H,K100E 19387 C117H,R122A 24257
E49A,R122E 9648 K105Y,R35F 14518 R88H,K101E 19388 C117H,R122E 24258
C117H,H124
E49A,H124A 9649 K105Y,R35G 14519 R88H,H102A 19389 24259
A
C117H,Q127
E49A,Q127A 9650 K105Y,R35I 14520 R88H,K105A 19390 24260
A
C117H,Q127
E49A,Q127K 9651 K105Y,R35L 14521 R88H,K105E 19391 24261
K
E49A,K128E 9652 K105Y,R35M 14522 R88H,K113A 19392 C117H,K128E 24262
R88H,S116
E49A,L133A 9653 K105Y,R35P 14523 19393 C117H,L133A 24263
W
E49A,L133V 9654 K105Y,R35S 14524 R88H,S116Y 19394 C117H,L133V 24264
P121A,K10E 9655 K105Y,R35V 14525 R88H,S116F 19395 C117L,K10E 24265
P121A,Y15A 9656 K105Y,D36A 14526 R88H,S116G 19396 C117L,Y15A 24266
P121A,Y15F 9657 K105Y,D36K 14527 R88H,S116N 19397 C117L,Y15F 24267
P121A,S17A 9658 K105Y,R37E 14528 R88H,S116K 19398 C117L,S17A 24268
P121A,S17K 9659 K105Y,R37A 14529 R88H,S116H 19399 C117L,S17K 24269
P121A,N18R 9660 K105Y,S38A 14530 R88H,S116Q 19400 C117L,N18R 24270
P121A,H21A 9661 K105Y,D39A 14531 R88H,C117K 19401 C117L,H21A 24271
P121A,L26A 9662 K105Y,Q40A 14532 R88H,C117R 19402 C117L,L26A 24272
P121A,P27A 9663 K105Y,Q43A 14533 R88H,C117S 19403 C117L,P27A 24273
P121A,D28K 9664 K105Y,Q43E 14534 R88H,C117Q 19404 C117L,D28K 24274
P121A,T30A 9665 K105Y,Q43K 14535 R88H,C117T 19405 C117L,T30A 24275
P121A,T34A 9666 K105Y,S47A 14536 R88H,C117P 19406 C117L,T34A 24276
P121A,R35A 9667 K105Y,S47V 14537 R88H,C117A 19407 C117L,R35A 24277
P121A,R35E 9668 K105Y,A48G 14538 R88H,C117I 19408 C117L,R35E 24278
R88H,K118
P121A,R35F 9669 K105Y,E53A 14539 19409 C117L,R35F 24279
W
P121A,R35G 9670 K105Y,Y55A 14540 R88H,K118H 19410 C117L,R35G 24280
P121A,R35I 9671 K105Y,Y55F 14541 R88H,K118R 19411 C117L,R35I 24281
P121A,R35L 9672 K105Y,K57A 14542 R88H,K118S 19412 C117L,R35L 24282
R88H,K118
P121A,R35M 9673 K105Y,K57E 14543 19413 C117L,R35M 24283
M
P121A,R35P 9674 K105Y,E60A 14544 R88H,K118L 19414 C117L,R35P 24284
P121A,R35S 9675 K105Y,E60K 14545 R88H,P121A 19415 C117L,R35S 24285
P121A,R35V 9676 K105Y,T61A 14546 R88H,R122A 19416 C117L,R35V 24286
Figure imgf000247_0001
Figure imgf000248_0001
P121A,R122
9750 N95D,R35G 14620 R88I,K105A 19490 C117M,K10E 24360 E
P121A,H124
9751 N95D,R35I 14621 R88I,K105E 19491 C117M,Y15A 24361 A
P121A,Q127
9752 N95D,R35L 14622 R88I,K113A 19492 C117M,Y15F 24362 A
P121A,Q127
9753 N95D,R35M 14623 R88I,S116W 19493 C117M,S17A 24363 K
P121A,K128
9754 N95D,R35P 14624 R88I,S116Y 19494 C117M,S17K 24364 E
P121A,L133
9755 N95D,R35S 14625 R88I,S116F 19495 C117M,N18R 24365 A
P121A,L133
9756 N95D,R35V 14626 R88I,S116G 19496 C117M,H21A 24366 V
Y15D,K10E 9757 N95D,D36A 14627 R88I,S116N 19497 C117M,L26A 24367
Y15D,S17A 9758 N95D,D36K 14628 R88I,S116K 19498 C117M,P27A 24368
Y15D,S17K 9759 N95D,R37E 14629 R88I,S116H 19499 C117M,D28K 24369
Y15D,N18R 9760 N95D,R37A 14630 R88I,S116Q 19500 C117M,T30A 24370
Y15D,H21A 9761 N95D,S38A 14631 R88I,C117K 19501 C117M,T34A 24371
Y15D,L26A 9762 N95D,D39A 14632 R88I,C117R 19502 C117M,R35A 24372
Y15D,P27A 9763 N95D,Q40A 14633 R88I,C117S 19503 C117M,R35E 24373
Y15D,D28K 9764 N95D,Q43A 14634 R88I,C117Q 19504 C117M,R35F 24374
Y15D,T30A 9765 N95D,Q43E 14635 R88I,C117T 19505 C117M,R35G 24375
Y15D,T34A 9766 N95D,Q43K 14636 R88I,C117P 19506 C117M,R35I 24376
Y15D,R35A 9767 N95D,S47A 14637 R88I,C117A 19507 C117M,R35L 24377
Y15D,R35E 9768 N95D,S47V 14638 R88I,C117I 19508 C117M,R35M 24378
Y15D,R35F 9769 N95D,A48G 14639 R88I,K118W 19509 C117M,R35P 24379
Y15D,R35G 9770 N95D,E53A 14640 R88I,K118H 19510 C117M,R35S 24380
Y15D,R35I 9771 N95D,Y55A 14641 R88I,K118R 19511 C117M,R35V 24381
Y15D,R35L 9772 N95D,Y55F 14642 R88I,K118S 19512 C117M,D36A 24382
Y15D,R35M 9773 N95D,K57A 14643 R88I,K118M 19513 C117M,D36K 24383
Y15D,R35P 9774 N95D,K57E 14644 R88I,K118L 19514 C117M,R37E 24384
Y15D,R35S 9775 N95D,E60A 14645 R88I,P121A 19515 C117M,R37A 24385
Y15D,R35V 9776 N95D,E60K 14646 R88I,R122A 19516 C117M,S38A 24386
Y15D,D36A 9777 N95D,T61A 14647 R88I,R122E 19517 C117M,D39A 24387
Y15D,D36K 9778 N95D,Q63A 14648 R88I,H124A 19518 C117M,Q40A 24388
Y15D,R37E 9779 N95D,Y64A 14649 R88I,Q127A 19519 C117M,Q43A 24389
Y15D,R37A 9780 N95D,D68A 14650 R88I,Q127K 19520 C117M,Q43E 24390
Y15D,S38A 9781 N95D,T78A 14651 R88I,K128E 19521 C117M,Q43K 24391
Y15D,D39A 9782 N95D,P79A 14652 R88I,L133A 19522 C117M,S47A 24392
Y15D,Q40A 9783 N95D,E81K 14653 R88I,L133V 19523 C117M,S47V 24393
Y15D,Q43A 9784 N95D,L84A 14654 R88K,K10E 19524 C117M,A48G 24394
Y15D,Q43E 9785 N95D,E87K 14655 R88K,Y15A 19525 C117M,E53A 24395
Y15D,Q43K 9786 N95D,E87Q 14656 R88K,Y15F 19526 C117M,Y55A 24396
Y15D,S47A 9787 N95D,E87V 14657 R88K,S17A 19527 C117M,Y55F 24397
Y15D,S47V 9788 N95D,E87D 14658 R88K,S17K 19528 C117M,K57A 24398
Y15D,A48G 9789 N95D,R88L 14659 R88K,N18R 19529 C117M,K57E 24399
Y15D,E53A 9790 N95D,R88Y 14660 R88K,H21A 19530 C117M,E60A 24400
Y15D,Y55A 9791 N95D,R88D 14661 R88K,L26A 19531 C117M,E60K 24401
Figure imgf000250_0001
W
C117M,K118
Y15D,S116N 9831 N95D,Q127A 14701 R88K,P79A 19571 24441
H
C117M,K118
Y15D,S116K 9832 N95D,Q127K 14702 R88K,E81K 19572 24442
R
C117M,K118
Y15D,S116H 9833 N95D,K128E 14703 R88K,L84A 19573 24443
S
C117M,K118
Y15D,S116Q 9834 N95D,L133A 14704 R88K,E87K 19574 24444
M
C117M,K118
Y15D,C117K 9835 N95D,L133V 14705 R88K,E87Q 19575 24445
L
C117M,P121
Y15D,C117R 9836 N95E,K10E 14706 R88K,E87V 19576 24446
A
C117M,R122
Y15D,C117S 9837 N95E,Y15A 14707 R88K,E87D 19577 24447
A
C117M,R122
Y15D,C117Q 9838 N95E,Y15F 14708 R88K,L89A 19578 24448
E
C117M,H124
Y15D,C117T 9839 N95E,S17A 14709 R88K,E90K 19579 24449
A
C117M,Q127
Y15D,C117P 9840 N95E,S17K 14710 R88K,E91A 19580 24450
A
C117M,Q127
Y15D,C117A 9841 N95E,N18R 14711 R88K,N92A 19581 24451
K
C117M,K128
Y15D,C117I 9842 N95E,H21A 14712 R88K,Y94V 19582 24452
E
Y15D,K118 C117M,L133
9843 N95E,L26A 14713 R88K,Y94A 19583 24453 W A
C117M,L133
Y15D,K118H 9844 N95E,P27A 14714 R88K,N95A 19584 24454
V
Y15D,K118R 9845 N95E,D28K 14715 R88K,N95V 19585 C117N,K10E 24455
Y15D,K118S 9846 N95E,T30A 14716 R88K,T96A 19586 C117N,Y15A 24456
Y15D,K118
9847 N95E,T34A 14717 R88K,K100E 19587 C117N,Y15F 24457 M
Y15D,K118L 9848 N95E,R35A 14718 R88K,K101E 19588 C117N,S17A 24458
Y15D,P121A 9849 N95E,R35E 14719 R88K,H102A 19589 C117N,S17K 24459
Y15D,R122A 9850 N95E,R35F 14720 R88K,K105A 19590 C117N,N18R 24460
Y15D,R122E 9851 N95E,R35G 14721 R88K,K105E 19591 C117N,H21A 24461
Y15D,H124A 9852 N95E,R35I 14722 R88K,K113A 19592 C117N,L26A 24462
R88K,S116
Y15D,Q127A 9853 N95E,R35L 14723 19593 C117N,P27A 24463
W
Y15D,Q127K 9854 N95E,R35M 14724 R88K,S116Y 19594 C117N,D28K 24464
Y15D,K128E 9855 N95E,R35P 14725 R88K,S116F 19595 C117N,T30A 24465
Y15D,L133A 9856 N95E,R35S 14726 R88K,S116G 19596 C117N,T34A 24466
Y15D,L133V 9857 N95E,R35V 14727 R88K,S116N 19597 C117N,R35A 24467
Y15E,K10E 9858 N95E,D36A 14728 R88K,S116K 19598 C117N,R35E 24468
Y15E,S17A 9859 N95E,D36K 14729 R88K,S116H 19599 C117N,R35F 24469
Y15E,S17K 9860 N95E,R37E 14730 R88K,S116Q 19600 C117N,R35G 24470
Y15E,N18R 9861 N95E,R37A 14731 R88K,C117K 19601 C117N,R35I 24471
Y15E,H21A 9862 N95E,S38A 14732 R88K,C117R 19602 C117N,R35L 24472
Y15E,L26A 9863 N95E,D39A 14733 R88K,C117S 19603 C117N,R35M 24473
Y15E,P27A 9864 N95E,Q40A 14734 R88K,C117Q 19604 C117N,R35P 24474
Y15E,D28K 9865 N95E,Q43A 14735 R88K,C117T 19605 C117N,R35S 24475
Υ15Ε/Γ30Α 9866 N95E,Q43E 14736 R88K,C117P 19606 C117N,R35V 24476 Υ15Ε/Γ34Α 9867 N95E,Q43K 14737 R88K,C117A 19607 C117N,D36A 24477
Y15E,R35A 9868 N95E,S47A 14738 R88K,C117I 19608 C117N,D36K 24478
R88K,K118
Y15E,R35E 9869 N95E,S47V 14739 19609 C117N,R37E 24479
W
Y15E,R35F 9870 N95E,A48G 14740 R88K,K118H 19610 C117N,R37A 24480
Y15E,R35G 9871 N95E,E53A 14741 R88K,K118R 19611 C117N,S38A 24481
Y15E,R35I 9872 N95E,Y55A 14742 R88K,K118S 19612 C117N,D39A 24482
R88K,K118
Y15E,R35L 9873 N95E,Y55F 14743 19613 C117N,Q40A 24483
M
Y15E,R35M 9874 N95E,K57A 14744 R88K,K118L 19614 C117N,Q43A 24484
Y15E,R35P 9875 N95E,K57E 14745 R88K,P121A 19615 C117N,Q43E 24485
Y15E,R35S 9876 N95E,E60A 14746 R88K,R122A 19616 C117N,Q43K 24486
Y15E,R35V 9877 N95E,E60K 14747 R88K,R122E 19617 C117N,S47A 24487
Y15E,D36A 9878 N95E,T61A 14748 R88K,H124A 19618 C117N,S47V 24488
Y15E,D36K 9879 N95E,Q63A 14749 R88K,Q127A 19619 C117N,A48G 24489
Y15E,R37E 9880 N95E,Y64A 14750 R88K,Q127K 19620 C117N,E53A 24490
Y15E,R37A 9881 N95E,D68A 14751 R88K,K128E 19621 C117N,Y55A 24491
Y15E,S38A 9882 N95E,T78A 14752 R88K,L133A 19622 C117N,Y55F 24492
Y15E,D39A 9883 N95E,P79A 14753 R88K,L133V 19623 C117N,K57A 24493
Y15E,Q40A 9884 N95E,E81K 14754 R88M,K10E 19624 C117N,K57E 24494
Y15E,Q43A 9885 N95E,L84A 14755 R88M,Y15A 19625 C117N,E60A 24495
Y15E,Q43E 9886 N95E,E87K 14756 R88M,Y15F 19626 C117N,E60K 24496
Y15E,Q43K 9887 N95E,E87Q 14757 R88M,S17A 19627 C117N,T61A 24497
Y15E,S47A 9888 N95E,E87V 14758 R88M,S17K 19628 C117N,Q63A 24498
Y15E,S47V 9889 N95E,E87D 14759 R88M,N18R 19629 C117N,Y64A 24499
Y15E,A48G 9890 N95E,R88L 14760 R88M,H21A 19630 C117N,D68A 24500
Y15E,E53A 9891 N95E,R88Y 14761 R88M,L26A 19631 C117N,T78A 24501
Y15E,Y55A 9892 N95E,R88D 14762 R88M,P27A 19632 C117N,P79A 24502
Y15E,Y55F 9893 N95E,L89A 14763 R88M,D28K 19633 C117N,E81K 24503
Y15E,K57A 9894 N95E,E90K 14764 R88M,T30A 19634 C117N,L84A 24504
Y15E,K57E 9895 N95E,E91A 14765 R88M,T34A 19635 C117N,E87K 24505
Y15E,E60A 9896 N95E,N92A 14766 R88M,R35A 19636 C117N,E87Q 24506
Y15E,E60K 9897 N95E,Y94V 14767 R88M,R35E 19637 C117N,E87V 24507
Y15E,T61A 9898 N95E,Y94A 14768 R88M,R35F 19638 C117N,E87D 24508
Y15E,Q63A 9899 N95E,T96A 14769 R88M,R35G 19639 C117N,R88L 24509
Y15E,Y64A 9900 N95E,K100E 14770 R88M,R35I 19640 C117N,R88Y 24510
Y15E,D68A 9901 N95E,K101E 14771 R88M,R35L 19641 C117N,R88D 24511
Y15E,T78A 9902 N95E,H102A 14772 R88M,R35M 19642 C117N,L89A 24512
Y15E,P79A 9903 N95E,K105A 14773 R88M,R35P 19643 C117N,E90K 24513
Y15E,E81K 9904 N95E,K105E 14774 R88M,R35S 19644 C117N,E91A 24514
Y15E,L84A 9905 N95E,K113A 14775 R88M,R35V 19645 C117N,N92A 24515
Y15E,E87K 9906 N95E,S116W 14776 R88M,D36A 19646 C117N,Y94V 24516
Y15E,E87Q 9907 N95E,S116Y 14777 R88M,D36K 19647 C117N,Y94A 24517
Y15E,E87V 9908 N95E,S116F 14778 R88M,R37E 19648 C117N,N95A 24518
Y15E,E87D 9909 N95E,S116G 14779 R88M,R37A 19649 C117N,N95V 24519
Y15E,R88L 9910 N95E,S116N 14780 R88M,S38A 19650 C117N,T96A 24520
Y15E,R88Y 9911 N95E,S116K 14781 R88M,D39A 19651 C117N,K100E 24521 Y15E,R88D 9912 N95E,S116H 14782 R88M,Q40A 19652 C117N,K101E 24522
C117N,H102
Y15E,L89A 9913 N95E,S116Q 14783 R88M,Q43A 19653 24523
A
C117N,K105
Y15E,E90K 9914 N95E,C117K 14784 R88M,Q43E 19654 24524
A
Y15E,E91A 9915 N95E,C117R 14785 R88M,Q43K 19655 C117N,K105E 24525
C117N,K113
Y15E,N92A 9916 N95E,C117S 14786 R88M,S47A 19656 24526
A
C117N,S116
Y15E,Y94V 9917 N95E,C117Q 14787 R88M,S47V 19657 24527
W
Y15E,Y94A 9918 N95E,C117T 14788 R88M,A48G 19658 C117N,S116Y 24528
Y15E,N95A 9919 N95E,C117P 14789 R88M,E53A 19659 C117N,S116F 24529
Y15E,N95V 9920 N95E,C117A 14790 R88M,Y55A 19660 C117N,S116G 24530
Y15E,T96A 9921 N95E,C117I 14791 R88M,Y55F 19661 C117N,S116N 24531
Y15E,K100E 9922 N95E,K118W 14792 R88M,K57A 19662 C117N,S116K 24532
Y15E,K101E 9923 N95E,K118H 14793 R88M,K57E 19663 C117N,S116H 24533
Y15E,H102A 9924 N95E,K118R 14794 R88M,E60A 19664 C117N,S116Q 24534
C117N,K118
Y15E,K105A 9925 N95E,K118S 14795 R88M,E60K 19665 24535
W
C117N,K118
Y15E,K105E 9926 N95E,K118M 14796 R88M,T61A 19666 24536
H
Y15E,K113A 9927 N95E,K118L 14797 R88M,Q63A 19667 C117N,K118R 24537
Y15E,S116W 9928 N95E,P121A 14798 R88M,Y64A 19668 C117N,K118S 24538
C117N,K118
Y15E,S116Y 9929 N95E,R122A 14799 R88M,D68A 19669 24539
M
Y15E,S116F 9930 N95E,R122E 14800 R88M,T78A 19670 C117N,K118L 24540
Y15E,S116G 9931 N95E,H124A 14801 R88M,P79A 19671 C117N,P121A 24541
Y15E,S116N 9932 N95E,Q127A 14802 R88M,E81K 19672 C117N,R122A 24542
Y15E,S116K 9933 N95E,Q127K 14803 R88M,L84A 19673 C117N,R122E 24543
C117N,H124
Y15E,S116H 9934 N95E,K128E 14804 R88M,E87K 19674 24544
A
C117N,Q127
Y15E,S116Q 9935 N95E,L133A 14805 R88M,E87Q 19675 24545
A
C117N,Q127
Y15E,C117K 9936 N95E,L133V 14806 R88M,E87V 19676 24546
K
Y15E,C117R 9937 N95F,K10E 14807 R88M,E87D 19677 C117N,K128E 24547
Y15E,C117S 9938 N95F,Y15A 14808 R88M,L89A 19678 C117N,L133A 24548
Y15E,C117Q 9939 N95F,Y15F 14809 R88M,E90K 19679 C117N,L133V 24549
Y15E,C117T 9940 N95F,S17A 14810 R88M,E91A 19680 C117W,K10E 24550
Y15E,C117P 9941 N95F,S17K 14811 R88M,N92A 19681 C117W,Y15A 24551
Y15E,C117A 9942 N95F,N18R 14812 R88M,Y94V 19682 C117W,Y15F 24552
Y15E,C117I 9943 N95F,H21A 14813 R88M,Y94A 19683 C117W,S17A 24553
Y15E,K118
9944 N95F,L26A 14814 R88M,N95A 19684 C117W,S17K 24554 W
Y15E,K118H 9945 N95F,P27A 14815 R88M,N95V 19685 C117W,N18R 24555
Y15E,K118R 9946 N95F,D28K 14816 R88M,T96A 19686 C117W,H21A 24556
R88M,K100
Y15E,K118S 9947 N95F,T30A 14817 19687 C117W,L26A 24557
E
R88M,K101
Y15E,K118M 9948 N95F,T34A 14818 19688 C117W,P27A 24558
E
R88M,H102
Y15E,K118L 9949 N95F,R35A 14819 19689 C117W,D28K 24559
A
Figure imgf000254_0001
Figure imgf000255_0001
R
C117W,K118
Y15G,K100E 10023 N95F,K118W 14893 R88N,K57E 19763 24633
S
C117W,K118
Y15G,K101E 10024 N95F,K118H 14894 R88N,E60A 19764 24634
M
C117W,K118
Y15G,H102A 10025 N95F,K118R 14895 R88N,E60K 19765 24635
L
C117W,P121
Y15G,K105A 10026 N95F,K118S 14896 R88N,T61A 19766 24636
A
C117W,R122
Y15G,K105E 10027 N95F,K118M 14897 R88N,Q63A 19767 24637
A
C117W,R122
Y15G,K113A 10028 N95F,K118L 14898 R88N,Y64A 19768 24638
E
C117W,H124
Y15G,S116W 10029 N95F,P121A 14899 R88N,D68A 19769 24639
A
C117W,Q127
Y15G,S116Y 10030 N95F,R122A 14900 R88N,T78A 19770 24640
A
C117W,Q127
Y15G,S116F 10031 N95F,R122E 14901 R88N,P79A 19771 24641
K
C117W,K128
Y15G,S116G 10032 N95F,H124A 14902 R88N,E81K 19772 24642
E
C117W,L133
Y15G,S116N 10033 N95F,Q127A 14903 R88N,L84A 19773 24643
A
C117W,L133
Y15G,S116K 10034 N95F,Q127K 14904 R88N,E87K 19774 24644
V
Y15G,S116H 10035 N95F,K128E 14905 R88N,E87Q 19775 C117Y,K10E 24645
Y15G,S116Q 10036 N95F,L133A 14906 R88N,E87V 19776 C117Y,Y15A 24646
Y15G,C117K 10037 N95F,L133V 14907 R88N,E87D 19777 C117Y,Y15F 24647
Y15G,C117R 10038 N95G,K10E 14908 R88N,L89A 19778 C117Y,S17A 24648
Y15G,C117S 10039 N95G,Y15A 14909 R88N,E90K 19779 C117Y,S17K 24649
Y15G,C117Q 10040 N95G,Y15F 14910 R88N,E91A 19780 C117Y,N18R 24650
Y15G,C117T 10041 N95G,S17A 14911 R88N,N92A 19781 C117Y,H21A 24651
Y15G,C117P 10042 N95G,S17K 14912 R88N,Y94V 19782 C117Y,L26A 24652
Y15G,C117A 10043 N95G,N18R 14913 R88N,Y94A 19783 C117Y,P27A 24653
Y15G,C117I 10044 N95G,H21A 14914 R88N,N95A 19784 C117Y,D28K 24654
Y15G,K118
10045 N95G,L26A 14915 R88N,N95V 19785 C117Y,T30A 24655 W
Y15G,K118H 10046 N95G,P27A 14916 R88N,T96A 19786 C117Y,T34A 24656
Y15G,K118R 10047 N95G,D28K 14917 R88N,K100E 19787 C117Y,R35A 24657
Y15G,K118S 10048 N95G,T30A 14918 R88N,K101E 19788 C117Y,R35E 24658
Y15G,K118
10049 N95G,T34A 14919 R88N,H102A 19789 C117Y,R35F 24659 M
Y15G,K118L 10050 N95G,R35A 14920 R88N,K105A 19790 C117Y,R35G 24660
Y15G,P121A 10051 N95G,R35E 14921 R88N,K105E 19791 C117Y,R35I 24661
Y15G,R122A 10052 N95G,R35F 14922 R88N,K113A 19792 C117Y,R35L 24662
R88N,S116
Y15G,R122E 10053 N95G,R35G 14923 19793 C117Y,R35M 24663
W
Y15G,H124A 10054 N95G,R35I 14924 R88N,S116Y 19794 C117Y,R35P 24664
Y15G,Q127A 10055 N95G,R35L 14925 R88N,S116F 19795 C117Y,R35S 24665
Y15G,Q127K 10056 N95G,R35M 14926 R88N,S116G 19796 C117Y,R35V 24666
Y15G,K128E 10057 N95G,R35P 14927 R88N,S116N 19797 C117Y,D36A 24667
Y15G,L133A 10058 N95G,R35S 14928 R88N,S116K 19798 C117Y,D36K 24668
Y15G,L133V 10059 N95G,R35V 14929 R88N,S116H 19799 C117Y,R37E 24669 Y15H,K10E 10060 N95G,D36A 14930 R88N,S116Q 19800 C117Y,R37A 24670
Y15H,S17A 10061 N95G,D36K 14931 R88N,C117K 19801 C117Y,S38A 24671
Y15H,S17K 10062 N95G,R37E 14932 R88N,C117R 19802 C117Y,D39A 24672
Y15H,N18R 10063 N95G,R37A 14933 R88N,C117S 19803 C117Y,Q40A 24673
Y15H,H21A 10064 N95G,S38A 14934 R88N,C117Q 19804 C117Y,Q43A 24674
Y15H,L26A 10065 N95G,D39A 14935 R88N,C117T 19805 C117Y,Q43E 24675
Y15H,P27A 10066 N95G,Q40A 14936 R88N,C117P 19806 C117Y,Q43K 24676
Y15H,D28K 10067 N95G,Q43A 14937 R88N,C117A 19807 C117Y,S47A 24677
Y15H,T30A 10068 N95G,Q43E 14938 R88N,C117I 19808 C117Y,S47V 24678
R88N,K118
Y15H,T34A 10069 N95G,Q43K 14939 19809 C117Y,A48G 24679
W
Y15H,R35A 10070 N95G,S47A 14940 R88N,K118H 19810 C117Y,E53A 24680
Y15H,R35E 10071 N95G,S47V 14941 R88N,K118R 19811 C117Y,Y55A 24681
Y15H,R35F 10072 N95G,A48G 14942 R88N,K118S 19812 C117Y,Y55F 24682
R88N,K118
Y15H,R35G 10073 N95G,E53A 14943 19813 C117Y,K57A 24683
M
Y15H,R35I 10074 N95G,Y55A 14944 R88N,K118L 19814 C117Y,K57E 24684
Y15H,R35L 10075 N95G,Y55F 14945 R88N,P121A 19815 C117Y,E60A 24685
Y15H,R35M 10076 N95G,K57A 14946 R88N,R122A 19816 C117Y,E60K 24686
Y15H,R35P 10077 N95G,K57E 14947 R88N,R122E 19817 C117Y,T61A 24687
Y15H,R35S 10078 N95G,E60A 14948 R88N,H124A 19818 C117Y,Q63A 24688
Y15H,R35V 10079 N95G,E60K 14949 R88N,Q127A 19819 C117Y,Y64A 24689
Y15H,D36A 10080 N95G,T61A 14950 R88N,Q127K 19820 C117Y,D68A 24690
Y15H,D36K 10081 N95G,Q63A 14951 R88N,K128E 19821 C117Y,T78A 24691
Y15H,R37E 10082 N95G,Y64A 14952 R88N,L133A 19822 C117Y,P79A 24692
Y15H,R37A 10083 N95G,D68A 14953 R88N,L133V 19823 C117Y,E81K 24693
Y15H,S38A 10084 N95G,T78A 14954 R88P,K10E 19824 C117Y,L84A 24694
Y15H,D39A 10085 N95G,P79A 14955 R88P,Y15A 19825 C117Y,E87K 24695
Y15H,Q40A 10086 N95G,E81K 14956 R88P,Y15F 19826 C117Y,E87Q 24696
Y15H,Q43A 10087 N95G,L84A 14957 R88P,S17A 19827 C117Y,E87V 24697
Y15H,Q43E 10088 N95G,E87K 14958 R88P,S17K 19828 C117Y,E87D 24698
Y15H,Q43K 10089 N95G,E87Q 14959 R88P,N18R 19829 C117Y,R88L 24699
Y15H,S47A 10090 N95G,E87V 14960 R88P,H21A 19830 C117Y,R88Y 24700
Y15H,S47V 10091 N95G,E87D 14961 R88P,L26A 19831 C117Y,R88D 24701
Y15H,A48G 10092 N95G,R88L 14962 R88P,P27A 19832 C117Y,L89A 24702
Y15H,E53A 10093 N95G,R88Y 14963 R88P,D28K 19833 C117Y,E90K 24703
Y15H,Y55A 10094 N95G,R88D 14964 R88P,T30A 19834 C117Y,E91A 24704
Y15H,Y55F 10095 N95G,L89A 14965 R88P,T34A 19835 C117Y,N92A 24705
Y15H,K57A 10096 N95G,E90K 14966 R88P,R35A 19836 C117Y,Y94V 24706
Y15H,K57E 10097 N95G,E91A 14967 R88P,R35E 19837 C117Y,Y94A 24707
Y15H,E60A 10098 N95G,N92A 14968 R88P,R35F 19838 C117Y,N95A 24708
Y15H,E60K 10099 N95G,Y94V 14969 R88P,R35G 19839 C117Y,N95V 24709
Y15H,T61A 10100 N95G,Y94A 14970 R88P,R35I 19840 C117Y,T96A 24710
Y15H,Q63A 10101 N95G,T96A 14971 R88P,R35L 19841 C117Y,K100E 24711
Y15H,Y64A 10102 N95G,K100E 14972 R88P,R35M 19842 C117Y,K101E 24712
C117Y,H102
Y15H,D68A 10103 N95G,K101E 14973 R88P,R35P 19843 24713
A
Y15H,T78A 10104 N95G,H102A 14974 R88P,R35S 19844 C117Y,K105 24714 A
Y15H,P79A 10105 N95G,K105A 14975 R88P,R35V 19845 C117Y,K105E 24715
C117Y,K113
Y15H,E81K 10106 N95G,K105E 14976 R88P,D36A 19846 24716
A
C117Y,S116
Y15H,L84A 10107 N95G,K113A 14977 R88P,D36K 19847 24717
W
Y15H,E87K 10108 N95G,S116W 14978 R88P,R37E 19848 C117Y,S116Y 24718
Y15H,E87Q 10109 N95G,S116Y 14979 R88P,R37A 19849 C117Y,S116F 24719
Y15H,E87V 10110 N95G,S116F 14980 R88P,S38A 19850 C117Y,S116G 24720
Y15H,E87D 10111 N95G,S116G 14981 R88P,D39A 19851 C117Y,S116N 24721
Y15H,R88L 10112 N95G,S116N 14982 R88P,Q40A 19852 C117Y,S116K 24722
Y15H,R88Y 10113 N95G,S116K 14983 R88P,Q43A 19853 C117Y,S116H 24723
Y15H,R88D 10114 N95G,S116H 14984 R88P,Q43E 19854 C117Y,S116Q 24724
C117Y,K118
Y15H,L89A 10115 N95G,S116Q 14985 R88P,Q43K 19855 24725
W
C117Y,K118
Y15H,E90K 10116 N95G,C117K 14986 R88P,S47A 19856 24726
H
Y15H,E91A 10117 N95G,C117R 14987 R88P,S47V 19857 C117Y,K118R 24727
Y15H,N92A 10118 N95G,C117S 14988 R88P,A48G 19858 C117Y,K118S 24728
C117Y,K118
Y15H,Y94V 10119 N95G,C117Q 14989 R88P,E53A 19859 24729
M
Y15H,Y94A 10120 N95G,C117T 14990 R88P,Y55A 19860 C117Y,K118L 24730
Y15H,N95A 10121 N95G,C117P 14991 R88P,Y55F 19861 C117Y,P121A 24731
Y15H,N95V 10122 N95G,C117A 14992 R88P,K57A 19862 C117Y,R122A 24732
Y15H,T96A 10123 N95G,C117I 14993 R88P,K57E 19863 C117Y,R122E 24733
C117Y,H124
Y15H,K100E 10124 N95G,K118W 14994 R88P,E60A 19864 24734
A
C117Y,Q127
Y15H,K101E 10125 N95G,K118H 14995 R88P,E60K 19865 24735
A
C117Y,Q127
Y15H,H102A 10126 N95G,K118R 14996 R88P,T61A 19866 24736
K
Y15H,K105A 10127 N95G,K118S 14997 R88P,Q63A 19867 C117Y,K128E 24737
Y15H,K105E 10128 N95G,K118M 14998 R88P,Y64A 19868 C117Y,L133A 24738
Y15H,K113A 10129 N95G,K118L 14999 R88P,D68A 19869 C117Y,L133V 24739
Y15H,S116W 10130 N95G,P121A 15000 R88P,T78A 19870 S116A,K10E 24740
Y15H,S116Y 10131 N95G,R122A 15001 R88P,P79A 19871 S116A,Y15A 24741
Y15H,S116F 10132 N95G,R122E 15002 R88P,E81K 19872 S116A,Y15F 24742
Y15H,S116G 10133 N95G,H124A 15003 R88P,L84A 19873 S116A,S17A 24743
Y15H,S116N 10134 N95G,Q127A 15004 R88P,E87K 19874 S116A,S17K 24744
Y15H,S116K 10135 N95G,Q127K 15005 R88P,E87Q 19875 S116A,N18R 24745
Y15H,S116H 10136 N95G,K128E 15006 R88P,E87V 19876 S116A,H21A 24746
Y15H,S116Q 10137 N95G,L133A 15007 R88P,E87D 19877 S116A,L26A 24747
Y15H,C117K 10138 N95G,L133V 15008 R88P,L89A 19878 S116A,P27A 24748
Y15H,C117R 10139 N95H,K10E 15009 R88P,E90K 19879 S116A,D28K 24749
Y15H,C117S 10140 N95H,Y15A 15010 R88P,E91A 19880 S116A,T30A 24750
Y15H,C117Q 10141 N95H,Y15F 15011 R88P,N92A 19881 S116A,T34A 24751
Y15H,C117T 10142 N95H,S17A 15012 R88P,Y94V 19882 S116A,R35A 24752
Y15H,C117P 10143 N95H,S17K 15013 R88P,Y94A 19883 S116A,R35E 24753
Y15H,C117A 10144 N95H,N18R 15014 R88P,N95A 19884 S116A,R35F 24754
Y15H,C117I 10145 N95H,H21A 15015 R88P,N95V 19885 S116A,R35G 24755 Y15H,K118
10146 N95H,L26A 15016 R88P,T96A 19886 S116A,R35I 24756 W
Y15H,K118H 10147 N95H,P27A 15017 R88P,K100E 19887 S116A,R35L 24757
Y15H,K118R 10148 N95H,D28K 15018 R88P,K101E 19888 S116A,R35M 24758
Y15H,K118S 10149 N95H,T30A 15019 R88P,H102A 19889 S116A,R35P 24759
Y15H,K118
10150 N95H,T34A 15020 R88P,K105A 19890 S116A,R35S 24760 M
Y15H,K118L 10151 N95H,R35A 15021 R88P,K105E 19891 S116A,R35V 24761
Y15H,P121A 10152 N95H,R35E 15022 R88P,K113A 19892 S116A,D36A 24762
Y15H,R122A 10153 N95H,R35F 15023 R88P,S116W 19893 S116A,D36K 24763
Y15H,R122E 10154 N95H,R35G 15024 R88P,S116Y 19894 S116A,R37E 24764
Y15H,H124A 10155 N95H,R35I 15025 R88P,S116F 19895 S116A,R37A 24765
Y15H,Q127A 10156 N95H,R35L 15026 R88P,S116G 19896 S116A,S38A 24766
Y15H,Q127K 10157 N95H,R35M 15027 R88P,S116N 19897 S116A,D39A 24767
Y15H,K128E 10158 N95H,R35P 15028 R88P,S116K 19898 S116A,Q40A 24768
Y15H,L133A 10159 N95H,R35S 15029 R88P,S116H 19899 S116A,Q43A 24769
Y15H,L133V 10160 N95H,R35V 15030 R88P,S116Q 19900 S116A,Q43E 24770
Y15I,K10E 10161 N95H,D36A 15031 R88P,C117K 19901 S116A,Q43K 24771
Y15I,S17A 10162 N95H,D36K 15032 R88P,C117R 19902 S116A,S47A 24772
Y15I,S17K 10163 N95H,R37E 15033 R88P,C117S 19903 S116A,S47V 24773
Y15I,N18R 10164 N95H,R37A 15034 R88P,C117Q 19904 S116A,A48G 24774
Y15I,H21A 10165 N95H,S38A 15035 R88P,C117T 19905 S116A,E53A 24775
Y15I,L26A 10166 N95H,D39A 15036 R88P,C117P 19906 S116A,Y55A 24776
Y15I,P27A 10167 N95H,Q40A 15037 R88P,C117A 19907 S116A,Y55F 24777
Y15I,D28K 10168 N95H,Q43A 15038 R88P,C117I 19908 S116A,K57A 24778
R88P,K118
Y15I,T30A 10169 N95H,Q43E 15039 19909 S116A,K57E 24779
W
Y15I,T34A 10170 N95H,Q43K 15040 R88P,K118H 19910 S116A,E60A 24780
Y15I,R35A 10171 N95H,S47A 15041 R88P,K118R 19911 S116A,E60K 24781
Y15I,R35E 10172 N95H,S47V 15042 R88P,K118S 19912 S116A,T61A 24782
Y15I,R35F 10173 N95H,A48G 15043 R88P,K118M 19913 S116A,Q63A 24783
Y15I,R35G 10174 N95H,E53A 15044 R88P,K118L 19914 S116A,Y64A 24784
Y15I,R35I 10175 N95H,Y55A 15045 R88P,P121A 19915 S116A,D68A 24785
Y15I,R35L 10176 N95H,Y55F 15046 R88P,R122A 19916 S116A,T78A 24786
Y15I,R35M 10177 N95H,K57A 15047 R88P,R122E 19917 S116A,P79A 24787
Y15I,R35P 10178 N95H,K57E 15048 R88P,H124A 19918 S116A,E81K 24788
Y15I,R35S 10179 N95H,E60A 15049 R88P,Q127A 19919 S116A,L84A 24789
Y15I,R35V 10180 N95H,E60K 15050 R88P,Q127K 19920 S116A,E87K 24790
Y15I,D36A 10181 Ν95Η/Γ61Α 15051 R88P,K128E 19921 S116A,E87Q 24791
Y15I,D36K 10182 N95H,Q63A 15052 R88P,L133A 19922 S116A,E87V 24792
Y15I,R37E 10183 N95H,Y64A 15053 R88P,L133V 19923 S116A,E87D 24793
Y15I,R37A 10184 N95H,D68A 15054 R88Q,K10E 19924 S116A,R88L 24794
Y15I,S38A 10185 N95H,T78A 15055 R88Q,Y15A 19925 S116A,R88Y 24795
Y15I,D39A 10186 N95H,P79A 15056 R88Q,Y15F 19926 S116A,R88D 24796
Y15I,Q40A 10187 N95H,E81K 15057 R88Q,S17A 19927 S116A,L89A 24797
Y15I,Q43A 10188 N95H,L84A 15058 R88Q,S17K 19928 S116A,E90K 24798
Y15I,Q43E 10189 N95H,E87K 15059 R88Q,N18R 19929 S116A,E91A 24799
Y15I,Q43K 10190 N95H,E87Q 15060 R88Q,H21A 19930 S116A,N92A 24800 Y15I,S47A 10191 N95H,E87V 15061 R88Q,L26A 19931 S116A,Y94V 24801
Y15I,S47V 10192 N95H,E87D 15062 R88Q,P27A 19932 S116A,Y94A 24802
Y15I,A48G 10193 N95H,R88L 15063 R88Q,D28K 19933 S116A,N95A 24803
Y15I,E53A 10194 N95H,R88Y 15064 R88Q,T30A 19934 S116A,N95V 24804
Y15I,Y55A 10195 N95H,R88D 15065 R88Q,T34A 19935 S116A,T96A 24805
Y15I,Y55F 10196 N95H,L89A 15066 R88Q,R35A 19936 S116A,K100E 24806
Y15I,K57A 10197 N95H,E90K 15067 R88Q,R35E 19937 S116A,K101E 24807
Y15I,K57E 10198 N95H,E91A 15068 R88Q,R35F 19938 S116A,H102A 24808
Y15I,E60A 10199 N95H,N92A 15069 R88Q,R35G 19939 S116A,K105A 24809
Y15I,E60K 10200 N95H,Y94V 15070 R88Q,R35I 19940 S116A,K105E 24810
Y15I,T61A 10201 N95H,Y94A 15071 R88Q,R35L 19941 S116A,K113A 24811
Y15I,Q63A 10202 N95H,T96A 15072 R88Q,R35M 19942 S116A,C117K 24812
Y15I,Y64A 10203 N95H,K100E 15073 R88Q,R35P 19943 S116A,C117R 24813
Y15I,D68A 10204 N95H,K101E 15074 R88Q,R35S 19944 S116A,C117S 24814
Y15I,T78A 10205 N95H,H102A 15075 R88Q,R35V 19945 S116A,C117Q 24815
Y15I,P79A 10206 N95H,K105A 15076 R88Q,D36A 19946 S116A,C117T 24816
Y15I,E81K 10207 N95H,K105E 15077 R88Q,D36K 19947 S116A,C117P 24817
Y15I,L84A 10208 N95H,K113A 15078 R88Q,R37E 19948 S116A,C117A 24818
Y15I,E87K 10209 N95H,S116W 15079 R88Q,R37A 19949 S116A,C117I 24819
S116A,K118
Y15I,E87Q 10210 N95H,S116Y 15080 R88Q,S38A 19950 24820
W
Y15I,E87V 10211 N95H,S116F 15081 R88Q,D39A 19951 S116A,K118H 24821
Y15I,E87D 10212 N95H,S116G 15082 R88Q,Q40A 19952 S116A,K118R 24822
Y15I,R88L 10213 N95H,S116N 15083 R88Q,Q43A 19953 S116A,K118S 24823
S116A,K118
Y15I,R88Y 10214 N95H,S116K 15084 R88Q,Q43E 19954 24824
M
Y15I,R88D 10215 N95H,S116H 15085 R88Q,Q43K 19955 S116A,K118L 24825
Y15I,L89A 10216 N95H,S116Q 15086 R88Q,S47A 19956 S116A,P121A 24826
Y15I,E90K 10217 N95H,C117K 15087 R88Q,S47V 19957 S116A,R122A 24827
Y15I,E91A 10218 N95H,C117R 15088 R88Q,A48G 19958 S116A,R122E 24828
Y15I,N92A 10219 N95H,C117S 15089 R88Q,E53A 19959 S116A,H124A 24829
Y15I,Y94V 10220 N95H,C117Q 15090 R88Q,Y55A 19960 S116A,Q127A 24830
Y15I,Y94A 10221 N95H,C117T 15091 R88Q,Y55F 19961 S116A,Q127K 24831
Y15I,N95A 10222 N95H,C117P 15092 R88Q,K57A 19962 S116A,K128E 24832
Y15I,N95V 10223 N95H,C117A 15093 R88Q,K57E 19963 S116A,L133A 24833
Y15I,T96A 10224 N95H,C117I 15094 R88Q,E60A 19964 S116A,L133V 24834
Y15I,K100E 10225 N95H,K118W 15095 R88Q,E60K 19965 S116D,K10E 24835
Y15I,K101E 10226 N95H,K118H 15096 R88Q,T61A 19966 S116D,Y15A 24836
Y15I,H102A 10227 N95H,K118R 15097 R88Q,Q63A 19967 S116D,Y15F 24837
Y15I,K105A 10228 N95H,K118S 15098 R88Q,Y64A 19968 S116D,S17A 24838
Y15I,K105E 10229 N95H,K118M 15099 R88Q,D68A 19969 S116D,S17K 24839
Y15I,K113A 10230 N95H,K118L 15100 R88Q,T78A 19970 S116D,N18R 24840
Y15I,S116W 10231 N95H,P121A 15101 R88Q,P79A 19971 S116D,H21A 24841
Y15I,S116Y 10232 N95H,R122A 15102 R88Q,E81K 19972 S116D,L26A 24842
Y15I,S116F 10233 N95H,R122E 15103 R88Q,L84A 19973 S116D,P27A 24843
Y15I,S116G 10234 N95H,H124A 15104 R88Q,E87K 19974 S116D,D28K 24844
Y15I,S116N 10235 N95H,Q127A 15105 R88Q,E87Q 19975 S116D,T30A 24845 Y15I,S116K 10236 N95H,Q127K 15106 R88Q,E87V 19976 S116D,T34A 24846
Y15I,S116H 10237 N95H,K128E 15107 R88Q,E87D 19977 S116D,R35A 24847
Y15I,S116Q 10238 N95H,L133A 15108 R88Q,L89A 19978 S116D,R35E 24848
Y15I,C117K 10239 N95H,L133V 15109 R88Q,E90K 19979 S116D,R35F 24849
Y15I,C117R 10240 N95I,K10E 15110 R88Q,E91A 19980 S116D,R35G 24850
Y15I,C117S 10241 N95LY15A 15111 R88Q,N92A 19981 S116D,R35I 24851
Y15I,C117Q 10242 N95I,Y15F 15112 R88Q,Y94V 19982 S116D,R35L 24852
Y15I,C117T 10243 N95LS17A 15113 R88Q,Y94A 19983 S116D,R35M 24853
Y15I,C117P 10244 N95I,S17K 15114 R88Q,N95A 19984 S116D,R35P 24854
Y15I,C117A 10245 N95I,N18R 15115 R88Q,N95V 19985 S116D,R35S 24855
Y15I,C117I 10246 N95I,H21A 15116 R88Q,T96A 19986 S116D,R35V 24856
Y15I,K118W 10247 N95I,L26A 15117 R88Q,K100E 19987 S116D,D36A 24857
Y15I,K118H 10248 N95I,P27A 15118 R88Q,K101E 19988 S116D,D36K 24858
Y15I,K118R 10249 N95I,D28K 15119 R88Q,H102A 19989 S116D,R37E 24859
Y15I,K118S 10250 N95I,T30A 15120 R88Q,K105A 19990 S116D,R37A 24860
Y15I,K118M 10251 N95I,T34A 15121 R88Q,K105E 19991 S116D,S38A 24861
Y15I,K118L 10252 N95I,R35A 15122 R88Q,K113A 19992 S116D,D39A 24862
R88Q,S116
Y15I,P121A 10253 N95I,R35E 15123 19993 S116D,Q40A 24863
W
Y15I,R122A 10254 N95I,R35F 15124 R88Q,S116Y 19994 S116D,Q43A 24864
Y15I,R122E 10255 N95I,R35G 15125 R88Q,S116F 19995 S116D,Q43E 24865
Y15I,H124A 10256 N95I,R35I 15126 R88Q,S116G 19996 S116D,Q43K 24866
Y15I,Q127A 10257 N95I,R35L 15127 R88Q,S116N 19997 S116D,S47A 24867
Y15I,Q127K 10258 N95I,R35M 15128 R88Q,S116K 19998 S116D,S47V 24868
Y15I,K128E 10259 N95I,R35P 15129 R88Q,S116H 19999 S116D,A48G 24869
Y15I,L133A 10260 N95I,R35S 15130 R88Q,S116Q 20000 S116D,E53A 24870
Y15I,L133V 10261 N95I,R35V 15131 R88Q,C117K 20001 S116D,Y55A 24871
Y15K,K10E 10262 N95I,D36A 15132 R88Q,C117R 20002 S116D,Y55F 24872
Y15K,S17A 10263 N95I,D36K 15133 R88Q,C117S 20003 S116D,K57A 24873
Y15K,S17K 10264 N95I,R37E 15134 R88Q,C117Q 20004 S116D,K57E 24874
Y15K,N18R 10265 N95I,R37A 15135 R88Q,C117T 20005 S116D,E60A 24875
Y15K,H21A 10266 N95I,S38A 15136 R88Q,C117P 20006 S116D,E60K 24876
Y15K,L26A 10267 N95I,D39A 15137 R88Q,C117A 20007 S116D,T61A 24877
Y15K,P27A 10268 N95I,Q40A 15138 R88Q,C117I 20008 S116D,Q63A 24878
R88Q,K118
Y15K,D28K 10269 N95I,Q43A 15139 20009 S116D,Y64A 24879
W
Y15K,T30A 10270 N95I,Q43E 15140 R88Q,K118H 20010 S116D,D68A 24880
Y15K,T34A 10271 N95I,Q43K 15141 R88Q,K118R 20011 S116D,T78A 24881
Y15K,R35A 10272 N95I,S47A 15142 R88Q,K118S 20012 S116D,P79A 24882
R88Q,K118
Y15K,R35E 10273 N95I,S47V 15143 20013 S116D,E81K 24883
M
Y15K,R35F 10274 N95I,A48G 15144 R88Q,K118L 20014 S116D,L84A 24884
Y15K,R35G 10275 N95I,E53A 15145 R88Q,P121A 20015 S116D,E87K 24885
Y15K,R35I 10276 N95I,Y55A 15146 R88Q,R122A 20016 S116D,E87Q 24886
Y15K,R35L 10277 N95I,Y55F 15147 R88Q,R122E 20017 S116D,E87V 24887
Y15K,R35M 10278 N95I,K57A 15148 R88Q,H124A 20018 S116D,E87D 24888
Y15K,R35P 10279 N95I,K57E 15149 R88Q,Q127A 20019 S116D,R88L 24889
Y15K,R35S 10280 N95I,E60A 15150 R88Q,Q127K 20020 S116D,R88Y 24890 Y15K,R35V 10281 N95I,E60K 15151 R88Q,K128E 20021 S116D,R88D 24891
Y15K,D36A 10282 Ν95Ι/Γ61Α 15152 R88Q,L133A 20022 S116D,L89A 24892
Y15K,D36K 10283 N95I,Q63A 15153 R88Q,L133V 20023 S116D,E90K 24893
Y15K,R37E 10284 N95I,Y64A 15154 R88S,K10E 20024 S116D,E91A 24894
Y15K,R37A 10285 N95I,D68A 15155 R88S,Y15A 20025 S116D,N92A 24895
Y15K,S38A 10286 N95I,T78A 15156 R88S,Y15F 20026 S116D,Y94V 24896
Y15K,D39A 10287 N95I,P79A 15157 R88S,S17A 20027 S116D,Y94A 24897
Y15K,Q40A 10288 N95I,E81K 15158 R88S,S17K 20028 S116D,N95A 24898
Y15K,Q43A 10289 N95I,L84A 15159 R88S,N18R 20029 S116D,N95V 24899
Y15K,Q43E 10290 N95I,E87K 15160 R88S,H21A 20030 S116D,T96A 24900
Y15K,Q43K 10291 N95I,E87Q 15161 R88S,L26A 20031 S116D,K100E 24901
Y15K,S47A 10292 N95I,E87V 15162 R88S,P27A 20032 S116D,K101E 24902
Y15K,S47V 10293 N95I,E87D 15163 R88S,D28K 20033 S116D,H102A 24903
Y15K,A48G 10294 N95I,R88L 15164 R88S,T30A 20034 S116D,K105A 24904
Y15K,E53A 10295 N95I,R88Y 15165 R88S,T34A 20035 S116D,K105E 24905
Y15K,Y55A 10296 N95I,R88D 15166 R88S,R35A 20036 S116D,K113A 24906
Y15K,Y55F 10297 N95I,L89A 15167 R88S,R35E 20037 S116D,C117K 24907
Y15K,K57A 10298 N95I,E90K 15168 R88S,R35F 20038 S116D,C117R 24908
Y15K,K57E 10299 N95I,E91A 15169 R88S,R35G 20039 S116D,C117S 24909
Y15K,E60A 10300 N95I,N92A 15170 R88S,R35I 20040 S116D,C117Q 24910
Y15K,E60K 10301 N95I,Y94V 15171 R88S,R35L 20041 S116D,C117T 24911
Y15K,T61A 10302 N95I,Y94A 15172 R88S,R35M 20042 S116D,C117P 24912
Y15K,Q63A 10303 N95I,T96A 15173 R88S,R35P 20043 S116D,C117A 24913
Y15K,Y64A 10304 N95I,K100E 15174 R88S,R35S 20044 S116D,C117I 24914
S116D,K118
Y15K,D68A 10305 N95I,K101E 15175 R88S,R35V 20045 24915
W
Y15K,T78A 10306 N95I,H102A 15176 R88S,D36A 20046 S116D,K118H 24916
Y15K,P79A 10307 N95I,K105A 15177 R88S,D36K 20047 S116D,K118R 24917
Y15K,E81K 10308 N95I,K105E 15178 R88S,R37E 20048 S116D,K118S 24918
S116D,K118
Y15K,L84A 10309 N95I,K113A 15179 R88S,R37A 20049 24919
M
Y15K,E87K 10310 N95I,S116W 15180 R88S,S38A 20050 S116D,K118L 24920
Y15K,E87Q 10311 N95I,S116Y 15181 R88S,D39A 20051 S116D,P121A 24921
Y15K,E87V 10312 N95I,S116F 15182 R88S,Q40A 20052 S116D,R122A 24922
Y15K,E87D 10313 N95I,S116G 15183 R88S,Q43A 20053 S116D,R122E 24923
Y15K,R88L 10314 N95I,S116N 15184 R88S,Q43E 20054 S116D,H124A 24924
Y15K,R88Y 10315 N95I,S116K 15185 R88S,Q43K 20055 S116D,Q127A 24925
Y15K,R88D 10316 N95I,S116H 15186 R88S,S47A 20056 S116D,Q127K 24926
Y15K,L89A 10317 N95I,S116Q 15187 R88S,S47V 20057 S116D,K128E 24927
Y15K,E90K 10318 N95I,C117K 15188 R88S,A48G 20058 S116D,L133A 24928
Y15K,E91A 10319 N95I,C117R 15189 R88S,E53A 20059 S116D,L133V 24929
Y15K,N92A 10320 N95I,C117S 15190 R88S,Y55A 20060 S116E,K10E 24930
Y15K,Y94V 10321 N95I,C117Q 15191 R88S,Y55F 20061 S116E,Y15A 24931
Y15K,Y94A 10322 N95I,C117T 15192 R88S,K57A 20062 S116E,Y15F 24932
Y15K,N95A 10323 N95I,C117P 15193 R88S,K57E 20063 S116E,S17A 24933
Y15K,N95V 10324 N95I,C117A 15194 R88S,E60A 20064 S116E,S17K 24934
Y15K,T96A 10325 N95I,C117I 15195 R88S,E60K 20065 S116E,N18R 24935 Y15K,K100E 10326 N95I,K118W 15196 R88S,T61A 20066 S116E,H21A 24936
Y15K,K101E 10327 N95I,K118H 15197 R88S,Q63A 20067 S116E,L26A 24937
Y15K,H102A 10328 N95I,K118R 15198 R88S,Y64A 20068 S116E,P27A 24938
Y15K,K105A 10329 N95I,K118S 15199 R88S,D68A 20069 S116E,D28K 24939
Y15K,K105E 10330 N95I,K118M 15200 R88S,T78A 20070 S116E,T30A 24940
Y15K,K113A 10331 N95I,K118L 15201 R88S,P79A 20071 S116E,T34A 24941
Y15K,S116W 10332 N95I,P121A 15202 R88S,E81K 20072 S116E,R35A 24942
Y15K,S116Y 10333 N95I,R122A 15203 R88S,L84A 20073 S116E,R35E 24943
Y15K,S116F 10334 N95I,R122E 15204 R88S,E87K 20074 S116E,R35F 24944
Y15K,S116G 10335 N95I,H124A 15205 R88S,E87Q 20075 S116E,R35G 24945
Y15K,S116N 10336 N95I,Q127A 15206 R88S,E87V 20076 S116E,R35I 24946
Y15K,S116K 10337 N95I,Q127K 15207 R88S,E87D 20077 S116E,R35L 24947
Y15K,S116H 10338 N95I,K128E 15208 R88S,L89A 20078 S116E,R35M 24948
Y15K,S116Q 10339 N95I,L133A 15209 R88S,E90K 20079 S116E,R35P 24949
Y15K,C117K 10340 N95I,L133V 15210 R88S,E91A 20080 S116E,R35S 24950
Y15K,C117R 10341 N95K,K10E 15211 R88S,N92A 20081 S116E,R35V 24951
Y15K,C117S 10342 N95K,Y15A 15212 R88S,Y94V 20082 S116E,D36A 24952
Y15K,C117Q 10343 N95K,Y15F 15213 R88S,Y94A 20083 S116E,D36K 24953
Y15K,C117T 10344 N95K,S17A 15214 R88S,N95A 20084 S116E,R37E 24954
Y15K,C117P 10345 N95K,S17K 15215 R88S,N95V 20085 S116E,R37A 24955
Y15K,C117A 10346 N95K,N18R 15216 R88S,T96A 20086 S116E,S38A 24956
Y15K,C117I 10347 N95K,H21A 15217 R88S,K100E 20087 S116E,D39A 24957
Y15K,K118
10348 N95K,L26A 15218 R88S,K101E 20088 S116E,Q40A 24958 W
Y15K,K118H 10349 N95K,P27A 15219 R88S,H102A 20089 S116E,Q43A 24959
Y15K,K118R 10350 N95K,D28K 15220 R88S,K105A 20090 S116E,Q43E 24960
Y15K,K118S 10351 N95K,T30A 15221 R88S,K105E 20091 S116E,Q43K 24961
Y15K,K118
10352 N95K,T34A 15222 R88S,K113A 20092 S116E,S47A 24962 M
Y15K,K118L 10353 N95K,R35A 15223 R88S,S116W 20093 S116E,S47V 24963
Y15K,P121A 10354 N95K,R35E 15224 R88S,S116Y 20094 S116E,A48G 24964
Y15K,R122A 10355 N95K,R35F 15225 R88S,S116F 20095 S116E,E53A 24965
Y15K,R122E 10356 N95K,R35G 15226 R88S,S116G 20096 S116E,Y55A 24966
Y15K,H124A 10357 N95K,R35I 15227 R88S,S116N 20097 S116E,Y55F 24967
Y15K,Q127A 10358 N95K,R35L 15228 R88S,S116K 20098 S116E,K57A 24968
Y15K,Q127K 10359 N95K,R35M 15229 R88S,S116H 20099 S116E,K57E 24969
Y15K,K128E 10360 N95K,R35P 15230 R88S,S116Q 20100 S116E,E60A 24970
Y15K,L133A 10361 N95K,R35S 15231 R88S,C117K 20101 S116E,E60K 24971
Y15K,L133V 10362 N95K,R35V 15232 R88S,C117R 20102 S116E,T61A 24972
Y15L,K10E 10363 N95K,D36A 15233 R88S,C117S 20103 S116E,Q63A 24973
Y15L,S17A 10364 N95K,D36K 15234 R88S,C117Q 20104 S116E,Y64A 24974
Y15L,S17K 10365 N95K,R37E 15235 R88S,C117T 20105 S116E,D68A 24975
Y15L,N18R 10366 N95K,R37A 15236 R88S,C117P 20106 S116E,T78A 24976
Y15L,H21A 10367 N95K,S38A 15237 R88S,C117A 20107 S116E,P79A 24977
Y15L,L26A 10368 N95K,D39A 15238 R88S,C117I 20108 S116E,E81K 24978
R88S,K118
Y15L,P27A 10369 N95K,Q40A 15239 20109 S116E,L84A 24979
W
Y15L,D28K 10370 N95K,Q43A 15240 R88S,K118H 20110 S116E,E87K 24980 Y15L,T30A 10371 N95K,Q43E 15241 R88S,K118R 20111 S116E,E87Q 24981
Y15L,T34A 10372 N95K,Q43K 15242 R88S,K118S 20112 S116E,E87V 24982
Y15L,R35A 10373 N95K,S47A 15243 R88S,K118M 20113 S116E,E87D 24983
Y15L,R35E 10374 N95K,S47V 15244 R88S,K118L 20114 S116E,R88L 24984
Y15L,R35F 10375 N95K,A48G 15245 R88S,P121A 20115 S116E,R88Y 24985
Y15L,R35G 10376 N95K,E53A 15246 R88S,R122A 20116 S116E,R88D 24986
Y15L,R35I 10377 N95K,Y55A 15247 R88S,R122E 20117 S116E,L89A 24987
Y15L,R35L 10378 N95K,Y55F 15248 R88S,H124A 20118 S116E,E90K 24988
Y15L,R35M 10379 N95K,K57A 15249 R88S,Q127A 20119 S116E,E91A 24989
Y15L,R35P 10380 N95K,K57E 15250 R88S,Q127K 20120 S116E,N92A 24990
Y15L,R35S 10381 N95K,E60A 15251 R88S,K128E 20121 S116E,Y94V 24991
Y15L,R35V 10382 N95K,E60K 15252 R88S,L133A 20122 S116E,Y94A 24992
Y15L,D36A 10383 N95K,T61A 15253 R88S,L133V 20123 S116E,N95A 24993
Y15L,D36K 10384 N95K,Q63A 15254 R88T,K10E 20124 S116E,N95V 24994
Y15L,R37E 10385 N95K,Y64A 15255 R88T,Y15A 20125 S116E,T96A 24995
Y15L,R37A 10386 N95K,D68A 15256 R88T,Y15F 20126 S116E,K100E 24996
Y15L,S38A 10387 N95K,T78A 15257 R88T,S17A 20127 S116E,K101E 24997
Y15L,D39A 10388 N95K,P79A 15258 R88T,S17K 20128 S116E,H102A 24998
Y15L,Q40A 10389 N95K,E81K 15259 R88T,N18R 20129 S116E,K105A 24999
Y15L,Q43A 10390 N95K,L84A 15260 R88T,H21A 20130 S116E,K105E 25000
Y15L,Q43E 10391 N95K,E87K 15261 R88T,L26A 20131 S116E,K113A 25001
Y15L,Q43K 10392 N95K,E87Q 15262 R88T,P27A 20132 S116E,C117K 25002
Y15L,S47A 10393 N95K,E87V 15263 R88T,D28K 20133 S116E,C117R 25003
Y15L,S47V 10394 N95K,E87D 15264 R88T,T30A 20134 S116E,C117S 25004
Y15L,A48G 10395 N95K,R88L 15265 R88T,T34A 20135 S116E,C117Q 25005
Y15L,E53A 10396 N95K,R88Y 15266 R88T,R35A 20136 S116E,C117T 25006
Y15L,Y55A 10397 N95K,R88D 15267 R88T,R35E 20137 S116E,C117P 25007
Y15L,Y55F 10398 N95K,L89A 15268 R88T,R35F 20138 S116E,C117A 25008
Y15L,K57A 10399 N95K,E90K 15269 R88T,R35G 20139 S116E,C117I 25009
S116E,K118
Y15L,K57E 10400 N95K,E91A 15270 R88T,R35I 20140 25010
W
Y15L,E60A 10401 N95K,N92A 15271 R88T,R35L 20141 S116E,K118H 25011
Y15L,E60K 10402 N95K,Y94V 15272 R88T,R35M 20142 S116E,K118R 25012
Y15L,T61A 10403 N95K,Y94A 15273 R88T,R35P 20143 S116E,K118S 25013
Y15L,Q63A 10404 N95K,T96A 15274 R88T,R35S 20144 S116E,K118M 25014
Y15L,Y64A 10405 N95K,K100E 15275 R88T,R35V 20145 S116E,K118L 25015
Y15L,D68A 10406 N95K,K101E 15276 R88T,D36A 20146 S116E,P121A 25016
Y15L,T78A 10407 N95K,H102A 15277 R88T,D36K 20147 S116E,R122A 25017
Y15L,P79A 10408 N95K,K105A 15278 R88T,R37E 20148 S116E,R122E 25018
Y15L,E81K 10409 N95K,K105E 15279 R88T,R37A 20149 S116E,H124A 25019
Y15L,L84A 10410 N95K,K113A 15280 R88T,S38A 20150 S116E,Q127A 25020
Y15L,E87K 10411 N95K,S116W 15281 R88T,D39A 20151 S116E,Q127K 25021
Y15L,E87Q 10412 N95K,S116Y 15282 R88T,Q40A 20152 S116E,K128E 25022
Y15L,E87V 10413 N95K,S116F 15283 R88T,Q43A 20153 S116E,L133A 25023
Y15L,E87D 10414 N95K,S116G 15284 R88T,Q43E 20154 S116E,L133V 25024
Y15L,R88L 10415 N95K,S116N 15285 R88T,Q43K 20155 S116I,K10E 25025
Y15L,R88Y 10416 N95K,S116K 15286 R88T,S47A 20156 S116I,Y15A 25026 Y15L,R88D 10417 N95K,S116H 15287 R88T,S47V 20157 S116I,Y15F 25027
Y15L,L89A 10418 N95K,S116Q 15288 R88T,A48G 20158 S116I,S17A 25028
Y15L,E90K 10419 N95K,C117K 15289 R88T,E53A 20159 S116I,S17K 25029
Y15L,E91A 10420 N95K,C117R 15290 R88T,Y55A 20160 S116I,N18R 25030
Y15L,N92A 10421 N95K,C117S 15291 R88T,Y55F 20161 S116I,H21A 25031
Y15L,Y94V 10422 N95K,C117Q 15292 R88T,K57A 20162 S116I,L26A 25032
Y15L,Y94A 10423 N95K,C117T 15293 R88T,K57E 20163 S116I,P27A 25033
Y15L,N95A 10424 N95K,C117P 15294 R88T,E60A 20164 S116I,D28K 25034
Y15L,N95V 10425 N95K,C117A 15295 R88T,E60K 20165 S116I,T30A 25035
Y15L,T96A 10426 N95K,C117I 15296 R88T,T61A 20166 S116I,T34A 25036
Y15L,K100E 10427 N95K,K118W 15297 R88T,Q63A 20167 S116I,R35A 25037
Y15L,K101E 10428 N95K,K118H 15298 R88T,Y64A 20168 S116I,R35E 25038
Y15L,H102A 10429 N95K,K118R 15299 R88T,D68A 20169 S116I,R35F 25039
Y15L,K105A 10430 N95K,K118S 15300 R88T,T78A 20170 S116I,R35G 25040
Y15L,K105E 10431 N95K,K118M 15301 R88T,P79A 20171 S116I,R35I 25041
Y15L,K113A 10432 N95K,K118L 15302 R88T,E81K 20172 S116I,R35L 25042
Y15L,S116W 10433 N95K,P121A 15303 R88T,L84A 20173 S116I,R35M 25043
Y15L,S116Y 10434 N95K,R122A 15304 R88T,E87K 20174 S116I,R35P 25044
Y15L,S116F 10435 N95K,R122E 15305 R88T,E87Q 20175 S116I,R35S 25045
Y15L,S116G 10436 N95K,H124A 15306 R88T,E87V 20176 S116I,R35V 25046
Y15L,S116N 10437 N95K,Q127A 15307 R88T,E87D 20177 S116I,D36A 25047
Y15L,S116K 10438 N95K,Q127K 15308 R88T,L89A 20178 S116I,D36K 25048
Y15L,S116H 10439 N95K,K128E 15309 R88T,E90K 20179 S116I,R37E 25049
Y15L,S116Q 10440 N95K,L133A 15310 R88T,E91A 20180 S116I,R37A 25050
Y15L,C117K 10441 N95K,L133V 15311 R88T,N92A 20181 S116I,S38A 25051
Y15L,C117R 10442 N95L,K10E 15312 R88T,Y94V 20182 S116I,D39A 25052
Y15L,C117S 10443 N95L,Y15A 15313 R88T,Y94A 20183 S116I,Q40A 25053
Y15L,C117Q 10444 N95L,Y15F 15314 R88T,N95A 20184 S116I,Q43A 25054
Y15L,C117T 10445 N95L,S17A 15315 R88T,N95V 20185 S116I,Q43E 25055
Y15L,C117P 10446 N95L,S17K 15316 R88T,T96A 20186 S116I,Q43K 25056
Y15L,C117A 10447 N95L,N18R 15317 R88T,K100E 20187 S116I,S47A 25057
Y15L,C117I 10448 N95L,H21A 15318 R88T,K101E 20188 S116I,S47V 25058
Y15L,K118
10449 N95L,L26A 15319 R88T,H102A 20189 S116I,A48G 25059 W
Y15L,K118H 10450 N95L,P27A 15320 R88T,K105A 20190 S116I,E53A 25060
Y15L,K118R 10451 N95L,D28K 15321 R88T,K105E 20191 S116I,Y55A 25061
Y15L,K118S 10452 N95L,T30A 15322 R88T,K113A 20192 S116I,Y55F 25062
Y15L,K118M 10453 N95L,T34A 15323 R88T,S116W 20193 S116I,K57A 25063
Y15L,K118L 10454 N95L,R35A 15324 R88T,S116Y 20194 S116I,K57E 25064
Y15L,P121A 10455 N95L,R35E 15325 R88T,S116F 20195 S116I,E60A 25065
Y15L,R122A 10456 N95L,R35F 15326 R88T,S116G 20196 S116I,E60K 25066
Y15L,R122E 10457 N95L,R35G 15327 R88T,S116N 20197 S116I,T61A 25067
Y15L,H124A 10458 N95L,R35I 15328 R88T,S116K 20198 S116I,Q63A 25068
Y15L,Q127A 10459 N95L,R35L 15329 R88T,S116H 20199 S116I,Y64A 25069
Y15L,Q127K 10460 N95L,R35M 15330 R88T,S116Q 20200 S116I,D68A 25070
Y15L,K128E 10461 N95L,R35P 15331 R88T,C117K 20201 S116I,T78A 25071
Y15L,L133A 10462 N95L,R35S 15332 R88T,C117R 20202 S116I,P79A 25072 Y15L,L133V 10463 N95L,R35V 15333 R88T,C117S 20203 S116I,E81K 25073
Y15M,K10E 10464 N95L,D36A 15334 R88T,C117Q 20204 S116I,L84A 25074
Y15M,S17A 10465 N95L,D36K 15335 R88T,C117T 20205 S116I,E87K 25075
Y15M,S17K 10466 N95L,R37E 15336 R88T,C117P 20206 S116I,E87Q 25076
Y15M,N18R 10467 N95L,R37A 15337 R88T,C117A 20207 S116I,E87V 25077
Y15M,H21A 10468 N95L,S38A 15338 R88T,C117I 20208 S116I,E87D 25078
R88T,K118
Y15M,L26A 10469 N95L,D39A 15339 20209 S116I,R88L 25079
W
Y15M,P27A 10470 N95L,Q40A 15340 R88T,K118H 20210 S116I,R88Y 25080
Y15M,D28K 10471 N95L,Q43A 15341 R88T,K118R 20211 S116I,R88D 25081
Y15M,T30A 10472 N95L,Q43E 15342 R88T,K118S 20212 S116I,L89A 25082
R88T,K118
Y15M,T34A 10473 N95L,Q43K 15343 20213 S116I,E90K 25083
M
Y15M,R35A 10474 N95L,S47A 15344 R88T,K118L 20214 S116I,E91A 25084
Y15M,R35E 10475 N95L,S47V 15345 R88T,P121A 20215 S116I,N92A 25085
Y15M,R35F 10476 N95L,A48G 15346 R88T,R122A 20216 S116I,Y94V 25086
Y15M,R35G 10477 N95L,E53A 15347 R88T,R122E 20217 S116I,Y94A 25087
Y15M,R35I 10478 N95L,Y55A 15348 R88T,H124A 20218 S116I,N95A 25088
Y15M,R35L 10479 N95L,Y55F 15349 R88T,Q127A 20219 S116I,N95V 25089
Y15M,R35M 10480 N95L,K57A 15350 R88T,Q127K 20220 S116I,T96A 25090
Y15M,R35P 10481 N95L,K57E 15351 R88T,K128E 20221 S116I,K100E 25091
Y15M,R35S 10482 N95L,E60A 15352 R88T,L133A 20222 S116I,K101E 25092
Y15M,R35V 10483 N95L,E60K 15353 R88T,L133V 20223 S116I,H102A 25093
Y15M,D36A 10484 N95L,T61A 15354 R88V,K10E 20224 S116I,K105A 25094
Y15M,D36K 10485 N95L,Q63A 15355 R88V,Y15A 20225 S116I,K105E 25095
Y15M,R37E 10486 N95L,Y64A 15356 R88V,Y15F 20226 S116I,K113A 25096
Y15M,R37A 10487 N95L,D68A 15357 R88V,S17A 20227 S116I,C117K 25097
Y15M,S38A 10488 N95L,T78A 15358 R88V,S17K 20228 S116I,C117R 25098
Y15M,D39A 10489 N95L,P79A 15359 R88V,N18R 20229 S116I,C117S 25099
Y15M,Q40A 10490 N95L,E81K 15360 R88V,H21A 20230 S116I,C117Q 25100
Y15M,Q43A 10491 N95L,L84A 15361 R88V,L26A 20231 S116I,C117T 25101
Y15M,Q43E 10492 N95L,E87K 15362 R88V,P27A 20232 S116I,C117P 25102
Y15M,Q43K 10493 N95L,E87Q 15363 R88V,D28K 20233 S116I,C117A 25103
Y15M,S47A 10494 N95L,E87V 15364 R88V,T30A 20234 S116I,C117I 25104
Y15M,S47V 10495 N95L,E87D 15365 R88V,T34A 20235 S116I,K118W 25105
Y15M,A48G 10496 N95L,R88L 15366 R88V,R35A 20236 S116I,K118H 25106
Y15M,E53A 10497 N95L,R88Y 15367 R88V,R35E 20237 S116I,K118R 25107
Y15M,Y55A 10498 N95L,R88D 15368 R88V,R35F 20238 S116I,K118S 25108
Y15M,Y55F 10499 N95L,L89A 15369 R88V,R35G 20239 S116I,K118M 25109
Y15M,K57A 10500 N95L,E90K 15370 R88V,R35I 20240 S116I,K118L 25110
Y15M,K57E 10501 N95L,E91A 15371 R88V,R35L 20241 S116I,P121A 25111
Y15M,E60A 10502 N95L,N92A 15372 R88V,R35M 20242 S116I,R122A 25112
Y15M,E60K 10503 N95L,Y94V 15373 R88V,R35P 20243 S116I,R122E 25113
Y15M,T61A 10504 N95L,Y94A 15374 R88V,R35S 20244 S116I,H124A 25114
Y15M,Q63A 10505 N95L,T96A 15375 R88V,R35V 20245 S116I,Q127A 25115
Y15M,Y64A 10506 N95L,K100E 15376 R88V,D36A 20246 S116I,Q127K 25116
Y15M,D68A 10507 N95L,K101E 15377 R88V,D36K 20247 S116I,K128E 25117
Figure imgf000267_0001
Y15M,K118
10550 N95M,L26A 15420 R88V,K105A 20290 S116L,E60A 25160 W
Y15M,K118
10551 N95M,P27A 15421 R88V,K105E 20291 S116L,E60K 25161 H
Y15M,K118
10552 N95M,D28K 15422 R88V,K113A 20292 S116L,T61A 25162 R
R88V,S116
Y15M,K118S 10553 N95M,T30A 15423 20293 S116L,Q63A 25163
W
Y15M,K118
10554 N95M,T34A 15424 R88V,S116Y 20294 S116L,Y64A 25164 M
Y15M,K118L 10555 N95M,R35A 15425 R88V,S116F 20295 S116L,D68A 25165
Y15M,P121A 10556 N95M,R35E 15426 R88V,S116G 20296 S116L,T78A 25166
Y15M,R122
10557 N95M,R35F 15427 R88V,S116N 20297 S116L,P79A 25167 A
Y15M,R122E 10558 N95M,R35G 15428 R88V,S116K 20298 S116L,E81K 25168
Y15M,H124
10559 N95M,R35I 15429 R88V,S116H 20299 S116L,L84A 25169 A
Y15M,Q127
10560 N95M,R35L 15430 R88V,S116Q 20300 S116L,E87K 25170 A
Y15M,Q127
10561 N95M,R35M 15431 R88V,C117K 20301 S116L,E87Q 25171 K
Y15M,K128E 10562 N95M,R35P 15432 R88V,C117R 20302 S116L,E87V 25172
Y15M,L133A 10563 N95M,R35S 15433 R88V,C117S 20303 S116L,E87D 25173
Y15M,L133V 10564 N95M,R35V 15434 R88V,C117Q 20304 S116L,R88L 25174
Y15N,K10E 10565 N95M,D36A 15435 R88V,C117T 20305 S116L,R88Y 25175
Y15N,S17A 10566 N95M,D36K 15436 R88V,C117P 20306 S116L,R88D 25176
Y15N,S17K 10567 N95M,R37E 15437 R88V,C117A 20307 S116L,L89A 25177
Y15N,N18R 10568 N95M,R37A 15438 R88V,C117I 20308 S116L,E90K 25178
R88V,K118
Y15N,H21A 10569 N95M,S38A 15439 20309 S116L,E91A 25179
W
Y15N,L26A 10570 N95M,D39A 15440 R88V,K118H 20310 S116L,N92A 25180
Y15N,P27A 10571 N95M,Q40A 15441 R88V,K118R 20311 S116L,Y94V 25181
Y15N,D28K 10572 N95M,Q43A 15442 R88V,K118S 20312 S116L,Y94A 25182
R88V,K118
Y15N,T30A 10573 N95M,Q43E 15443 20313 S116L,N95A 25183
M
Υ15Ν/Γ34Α 10574 N95M,Q43K 15444 R88V,K118L 20314 S116L,N95V 25184
Y15N,R35A 10575 N95M,S47A 15445 R88V,P121A 20315 S116L,T96A 25185
Y15N,R35E 10576 N95M,S47V 15446 R88V,R122A 20316 S116L,K100E 25186
Y15N,R35F 10577 N95M,A48G 15447 R88V,R122E 20317 S116L,K101E 25187
Y15N,R35G 10578 N95M,E53A 15448 R88V,H124A 20318 S116L,H102A 25188
Y15N,R35I 10579 N95M,Y55A 15449 R88V,Q127A 20319 S116L,K105A 25189
Y15N,R35L 10580 N95M,Y55F 15450 R88V,Q127K 20320 S116L,K105E 25190
Y15N,R35M 10581 N95M,K57A 15451 R88V,K128E 20321 S116L,K113A 25191
Y15N,R35P 10582 N95M,K57E 15452 R88V,L133A 20322 S116L,C117K 25192
Y15N,R35S 10583 N95M,E60A 15453 R88V,L133V 20323 S116L,C117R 25193
Y15N,R35V 10584 N95M,E60K 15454 R88W,K10E 20324 S116L,C117S 25194
Y15N,D36A 10585 N95M,T61A 15455 R88W,Y15A 20325 S116L,C117Q 25195
Y15N,D36K 10586 N95M,Q63A 15456 R88W,Y15F 20326 S116L,C117T 25196
Y15N,R37E 10587 N95M,Y64A 15457 R88W,S17A 20327 S116L,C117P 25197
Y15N,R37A 10588 N95M,D68A 15458 R88W,S17K 20328 S116L,C117A 25198
Y15N,S38A 10589 N95M,T78A 15459 R88W,N18R 20329 S116L,C117I 25199 S116L,K118
Y15N,D39A 10590 N95M,P79A 15460 R88W,H21A 20330 25200
W
Y15N,Q40A 10591 N95M,E81K 15461 R88W,L26A 20331 S116L,K118H 25201
Y15N,Q43A 10592 N95M,L84A 15462 R88W,P27A 20332 S116L,K118R 25202
Y15N,Q43E 10593 N95M,E87K 15463 R88W,D28K 20333 S116L,K118S 25203
Y15N,Q43K 10594 N95M,E87Q 15464 R88W,T30A 20334 S116L,K118M 25204
Y15N,S47A 10595 N95M,E87V 15465 R88W,T34A 20335 S116L,K118L 25205
Y15N,S47V 10596 N95M,E87D 15466 R88W,R35A 20336 S116L,P121A 25206
Y15N,A48G 10597 N95M,R88L 15467 R88W,R35E 20337 S116L,R122A 25207
Y15N,E53A 10598 N95M,R88Y 15468 R88W,R35F 20338 S116L,R122E 25208
Y15N,Y55A 10599 N95M,R88D 15469 R88W,R35G 20339 S116L,H124A 25209
Y15N,Y55F 10600 N95M,L89A 15470 R88W,R35I 20340 S116L,Q127A 25210
Y15N,K57A 10601 N95M,E90K 15471 R88W,R35L 20341 S116L,Q127K 25211
Y15N,K57E 10602 N95M,E91A 15472 R88W,R35M 20342 S116L,K128E 25212
Y15N,E60A 10603 N95M,N92A 15473 R88W,R35P 20343 S116L,L133A 25213
Y15N,E60K 10604 N95M,Y94V 15474 R88W,R35S 20344 S116L,L133V 25214
Y15N,T61A 10605 N95M,Y94A 15475 R88W,R35V 20345 S116M,K10E 25215
Y15N,Q63A 10606 N95M,T96A 15476 R88W,D36A 20346 S116M,Y15A 25216
Y15N,Y64A 10607 N95M,K100E 15477 R88W,D36K 20347 S116M,Y15F 25217
Y15N,D68A 10608 N95M,K101E 15478 R88W,R37E 20348 S116M,S17A 25218
Y15N,T78A 10609 N95M,H102A 15479 R88W,R37A 20349 S116M,S17K 25219
Y15N,P79A 10610 N95M,K105A 15480 R88W,S38A 20350 S116M,N18R 25220
Y15N,E81K 10611 N95M,K105E 15481 R88W,D39A 20351 S116M,H21A 25221
Y15N,L84A 10612 N95M,K113A 15482 R88W,Q40A 20352 S116M,L26A 25222
Y15N,E87K 10613 N95M,S116W 15483 R88W,Q43A 20353 S116M,P27A 25223
Y15N,E87Q 10614 N95M,S116Y 15484 R88W,Q43E 20354 S116M,D28K 25224
Y15N,E87V 10615 N95M,S116F 15485 R88W,Q43K 20355 S116M,T30A 25225
Y15N,E87D 10616 N95M,S116G 15486 R88W,S47A 20356 S116M,T34A 25226
Y15N,R88L 10617 N95M,S116N 15487 R88W,S47V 20357 S116M,R35A 25227
Y15N,R88Y 10618 N95M,S116K 15488 R88W,A48G 20358 S116M,R35E 25228
Y15N,R88D 10619 N95M,S116H 15489 R88W,E53A 20359 S116M,R35F 25229
Y15N,L89A 10620 N95M,S116Q 15490 R88W,Y55A 20360 S116M,R35G 25230
Y15N,E90K 10621 N95M,C117K 15491 R88W,Y55F 20361 S116M,R35I 25231
Y15N,E91A 10622 N95M,C117R 15492 R88W,K57A 20362 S116M,R35L 25232
Y15N,N92A 10623 N95M,C117S 15493 R88W,K57E 20363 S116M,R35M 25233
Y15N,Y94V 10624 N95M,C117Q 15494 R88W,E60A 20364 S116M,R35P 25234
Y15N,Y94A 10625 N95M,C117T 15495 R88W,E60K 20365 S116M,R35S 25235
Y15N,N95A 10626 N95M,C117P 15496 R88W,T61A 20366 S116M,R35V 25236
Y15N,N95V 10627 N95M,C117A 15497 R88W,Q63A 20367 S116M,D36A 25237
Y15N,T96A 10628 N95M,C117I 15498 R88W,Y64A 20368 S116M,D36K 25238
Y15N,K100E 10629 N95M,K118W 15499 R88W,D68A 20369 S116M,R37E 25239
Y15N,K101E 10630 N95M,K118H 15500 R88W,T78A 20370 S116M,R37A 25240
Y15N,H102A 10631 N95M,K118R 15501 R88W,P79A 20371 S116M,S38A 25241
Y15N,K105A 10632 N95M,K118S 15502 R88W,E81K 20372 S116M,D39A 25242
Y15N,K105E 10633 N95M,K118M 15503 R88W,L84A 20373 S116M,Q40A 25243
Y15N,K113A 10634 N95M,K118L 15504 R88W,E87K 20374 S116M,Q43A 25244
Y15N,S116W 10635 N95M,P121A 15505 R88W,E87Q 20375 S116M,Q43E 25245
Figure imgf000270_0001
Figure imgf000271_0001
Y15P,E60A 10704 N95P,N92A 15574 L84D,R35S 20444 S116P,S17K 25314
Y15P,E60K 10705 N95P,Y94V 15575 L84D,R35V 20445 S116P,N18R 25315
Y15P,T61A 10706 N95P,Y94A 15576 L84D,D36A 20446 S116P,H21A 25316
Y15P,Q63A 10707 N95P,T96A 15577 L84D,D36K 20447 S116P,L26A 25317
Y15P,Y64A 10708 N95P,K100E 15578 L84D,R37E 20448 S116P,P27A 25318
Y15P,D68A 10709 N95P,K101E 15579 L84D,R37A 20449 S116P,D28K 25319
Y15P,T78A 10710 N95P,H102A 15580 L84D,S38A 20450 S116P,T30A 25320
Y15P,P79A 10711 N95P,K105A 15581 L84D,D39A 20451 S116P,T34A 25321
Y15P,E81K 10712 N95P,K105E 15582 L84D,Q40A 20452 S116P,R35A 25322
Y15P,L84A 10713 N95P,K113A 15583 L84D,Q43A 20453 S116P,R35E 25323
Y15P,E87K 10714 N95P,S116W 15584 L84D,Q43E 20454 S116P,R35F 25324
Y15P,E87Q 10715 N95P,S116Y 15585 L84D,Q43K 20455 S116P,R35G 25325
Y15P,E87V 10716 N95P,S116F 15586 L84D,S47A 20456 S116P,R35I 25326
Y15P,E87D 10717 N95P,S116G 15587 L84D,S47V 20457 S116P,R35L 25327
Y15P,R88L 10718 N95P,S116N 15588 L84D,A48G 20458 S116P,R35M 25328
Y15P,R88Y 10719 N95P,S116K 15589 L84D,E53A 20459 S116P,R35P 25329
Y15P,R88D 10720 N95P,S116H 15590 L84D,Y55A 20460 S116P,R35S 25330
Y15P,L89A 10721 N95P,S116Q 15591 L84D,Y55F 20461 S116P,R35V 25331
Y15P,E90K 10722 N95P,C117K 15592 L84D,K57A 20462 S116P,D36A 25332
Y15P,E91A 10723 N95P,C117R 15593 L84D,K57E 20463 S116P,D36K 25333
Y15P,N92A 10724 N95P,C117S 15594 L84D,E60A 20464 S116P,R37E 25334
Y15P,Y94V 10725 N95P,C117Q 15595 L84D,E60K 20465 S116P,R37A 25335
Y15P,Y94A 10726 N95P,C117T 15596 L84D,T61A 20466 S116P,S38A 25336
Y15P,N95A 10727 N95P,C117P 15597 L84D,Q63A 20467 S116P,D39A 25337
Y15P,N95V 10728 N95P,C117A 15598 L84D,Y64A 20468 S116P,Q40A 25338
Y15P,T96A 10729 N95P,C117I 15599 L84D,D68A 20469 S116P,Q43A 25339
Y15P,K100E 10730 N95P,K118W 15600 L84D,T78A 20470 S116P,Q43E 25340
Y15P,K101E 10731 N95P,K118H 15601 L84D,P79A 20471 S116P,Q43K 25341
Y15P,H102A 10732 N95P,K118R 15602 L84D,E81K 20472 S116P,S47A 25342
Y15P,K105A 10733 N95P,K118S 15603 L84D,E87K 20473 S116P,S47V 25343
Y15P,K105E 10734 N95P,K118M 15604 L84D,E87Q 20474 S116P,A48G 25344
Y15P,K113A 10735 N95P,K118L 15605 L84D,E87V 20475 S116P,E53A 25345
Y15P,S116W 10736 N95P,P121A 15606 L84D,E87D 20476 S116P,Y55A 25346
Y15P,S116Y 10737 N95P,R122A 15607 L84D,R88L 20477 S116P,Y55F 25347
Y15P,S116F 10738 N95P,R122E 15608 L84D,R88Y 20478 S116P,K57A 25348
Y15P,S116G 10739 N95P,H124A 15609 L84D,R88D 20479 S116P,K57E 25349
Y15P,S116N 10740 N95P,Q127A 15610 L84D,L89A 20480 S116P,E60A 25350
Y15P,S116K 10741 N95P,Q127K 15611 L84D,E90K 20481 S116P,E60K 25351
Y15P,S116H 10742 N95P,K128E 15612 L84D,E91A 20482 S116P,T61A 25352
Y15P,S116Q 10743 N95P,L133A 15613 L84D,N92A 20483 S116P,Q63A 25353
Y15P,C117K 10744 N95P,L133V 15614 L84D,Y94V 20484 S116P,Y64A 25354
Y15P,C117R 10745 N95Q,K10E 15615 L84D,Y94A 20485 S116P,D68A 25355
Y15P,C117S 10746 N95Q,Y15A 15616 L84D,N95A 20486 S116P,T78A 25356
Y15P,C117Q 10747 N95Q,Y15F 15617 L84D,N95V 20487 S116P,P79A 25357
Y15P,C117T 10748 N95Q,S17A 15618 L84D,T96A 20488 S116P,E81K 25358
Y15P,C117P 10749 N95Q,S17K 15619 L84D,K100E 20489 S116P,L84A 25359 Y15P,C117A 10750 N95Q,N18R 15620 L84D,K101E 20490 S116P,E87K 25360
Y15P,C117I 10751 N95Q,H21A 15621 L84D,H102A 20491 S116P,E87Q 25361
Y15P,K118W 10752 N95Q,L26A 15622 L84D,K105A 20492 S116P,E87V 25362
Y15P,K118H 10753 N95Q,P27A 15623 L84D,K105E 20493 S116P,E87D 25363
Y15P,K118R 10754 N95Q,D28K 15624 L84D,K113A 20494 S116P,R88L 25364
Y15P,K118S 10755 N95Q,T30A 15625 L84D,S116W 20495 S116P,R88Y 25365
Y15P,K118M 10756 N95Q,T34A 15626 L84D,S116Y 20496 S116P,R88D 25366
Y15P,K118L 10757 N95Q,R35A 15627 L84D,S116F 20497 S116P,L89A 25367
Y15P,P121A 10758 N95Q,R35E 15628 L84D,S116G 20498 S116P,E90K 25368
Y15P,R122A 10759 N95Q,R35F 15629 L84D,S116N 20499 S116P,E91A 25369
Y15P,R122E 10760 N95Q,R35G 15630 L84D,S116K 20500 S116P,N92A 25370
Y15P,H124A 10761 N95Q,R35I 15631 L84D,S116H 20501 S116P,Y94V 25371
Y15P,Q127A 10762 N95Q,R35L 15632 L84D,S116Q 20502 S116P,Y94A 25372
Y15P,Q127K 10763 N95Q,R35M 15633 L84D,C117K 20503 S116P,N95A 25373
Y15P,K128E 10764 N95Q,R35P 15634 L84D,C117R 20504 S116P,N95V 25374
Y15P,L133A 10765 N95Q,R35S 15635 L84D,C117S 20505 S116P,T96A 25375
Y15P,L133V 10766 N95Q,R35V 15636 L84D,C117Q 20506 S116P,K100E 25376
Y15Q,K10E 10767 N95Q,D36A 15637 L84D,C117T 20507 S116P,K101E 25377
Y15Q,S17A 10768 N95Q,D36K 15638 L84D,C117P 20508 S116P,H102A 25378
Y15Q,S17K 10769 N95Q,R37E 15639 L84D,C117A 20509 S116P,K105A 25379
Y15Q,N18R 10770 N95Q,R37A 15640 L84D,C117I 20510 S116P,K105E 25380
L84D,K118
Y15Q,H21A 10771 N95Q,S38A 15641 20511 S116P,K113A 25381
W
Y15Q,L26A 10772 N95Q,D39A 15642 L84D,K118H 20512 S116P,C117K 25382
Y15Q,P27A 10773 N95Q,Q40A 15643 L84D,K118R 20513 S116P,C117R 25383
Y15Q,D28K 10774 N95Q,Q43A 15644 L84D,K118S 20514 S116P,C117S 25384
L84D,K118
Y15Q,T30A 10775 N95Q,Q43E 15645 20515 S116P,C117Q 25385
M
Y15Q,T34A 10776 N95Q,Q43K 15646 L84D,K118L 20516 S116P,C117T 25386
Y15Q,R35A 10777 N95Q,S47A 15647 L84D,P121A 20517 S116P,C117P 25387
Y15Q,R35E 10778 N95Q,S47V 15648 L84D,R122A 20518 S116P,C117A 25388
Y15Q,R35F 10779 N95Q,A48G 15649 L84D,R122E 20519 S116P,C117I 25389
Y15Q,R35G 10780 N95Q,E53A 15650 L84D,H124A 20520 S116P,K118W 25390
Y15Q,R35I 10781 N95Q,Y55A 15651 L84D,Q127A 20521 S116P,K118H 25391
Y15Q,R35L 10782 N95Q,Y55F 15652 L84D,Q127K 20522 S116P,K118R 25392
Y15Q,R35M 10783 N95Q,K57A 15653 L84D,K128E 20523 S116P,K118S 25393
Y15Q,R35P 10784 N95Q,K57E 15654 L84D,L133A 20524 S116P,K118M 25394
Y15Q,R35S 10785 N95Q,E60A 15655 L84D,L133V 20525 S116P,K118L 25395
Y15Q,R35V 10786 N95Q,E60K 15656 L84E,K10E 20526 S116P,P121A 25396
Y15Q,D36A 10787 N95Q,T61A 15657 L84E,Y15A 20527 S116P,R122A 25397
Y15Q,D36K 10788 N95Q,Q63A 15658 L84E,Y15F 20528 S116P,R122E 25398
Y15Q,R37E 10789 N95Q,Y64A 15659 L84E,S17A 20529 S116P,H124A 25399
Y15Q,R37A 10790 N95Q,D68A 15660 L84E,S17K 20530 S116P,Q127A 25400
Y15Q,S38A 10791 N95Q,T78A 15661 L84E,N18R 20531 S116P,Q127K 25401
Y15Q,D39A 10792 N95Q,P79A 15662 L84E,H21A 20532 S116P,K128E 25402
Y15Q,Q40A 10793 N95Q,E81K 15663 L84E,L26A 20533 S116P,L133A 25403
Y15Q,Q43A 10794 N95Q,L84A 15664 L84E,P27A 20534 S116P,L133V 25404 Y15Q,Q43E 10795 N95Q,E87K 15665 L84E,D28K 20535 S116T,K10E 25405
Y15Q,Q43K 10796 N95Q,E87Q 15666 L84E,T30A 20536 S116T,Y15A 25406
Y15Q,S47A 10797 N95Q,E87V 15667 L84E,T34A 20537 S116T,Y15F 25407
Y15Q,S47V 10798 N95Q,E87D 15668 L84E,R35A 20538 S116T,S17A 25408
Y15Q,A48G 10799 N95Q,R88L 15669 L84E,R35E 20539 S116T,S17K 25409
Y15Q,E53A 10800 N95Q,R88Y 15670 L84E,R35F 20540 S116T,N18R 25410
Y15Q,Y55A 10801 N95Q,R88D 15671 L84E,R35G 20541 S116T,H21A 25411
Y15Q,Y55F 10802 N95Q,L89A 15672 L84E,R35I 20542 S116T,L26A 25412
Y15Q,K57A 10803 N95Q,E90K 15673 L84E,R35L 20543 S116T,P27A 25413
Y15Q,K57E 10804 N95Q,E91A 15674 L84E,R35M 20544 S116T,D28K 25414
Y15Q,E60A 10805 N95Q,N92A 15675 L84E,R35P 20545 S116T,T30A 25415
Y15Q,E60K 10806 N95Q,Y94V 15676 L84E,R35S 20546 S116T,T34A 25416
Y15Q,T61A 10807 N95Q,Y94A 15677 L84E,R35V 20547 S116T,R35A 25417
Y15Q,Q63A 10808 N95Q,T96A 15678 L84E,D36A 20548 S116T,R35E 25418
Y15Q,Y64A 10809 N95Q,K100E 15679 L84E,D36K 20549 S116T,R35F 25419
Y15Q,D68A 10810 N95Q,K101E 15680 L84E,R37E 20550 S116T,R35G 25420
Y15Q,T78A 10811 N95Q,H102A 15681 L84E,R37A 20551 S116T,R35I 25421
Y15Q,P79A 10812 N95Q,K105A 15682 L84E,S38A 20552 S116T,R35L 25422
Y15Q,E81K 10813 N95Q,K105E 15683 L84E,D39A 20553 S116T,R35M 25423
Y15Q,L84A 10814 N95Q,K113A 15684 L84E,Q40A 20554 S116T,R35P 25424
Y15Q,E87K 10815 N95Q,S116W 15685 L84E,Q43A 20555 S116T,R35S 25425
Y15Q,E87Q 10816 N95Q,S116Y 15686 L84E,Q43E 20556 S116T,R35V 25426
Y15Q,E87V 10817 N95Q,S116F 15687 L84E,Q43K 20557 S116T,D36A 25427
Y15Q,E87D 10818 N95Q,S116G 15688 L84E,S47A 20558 S116T,D36K 25428
Y15Q,R88L 10819 N95Q,S116N 15689 L84E,S47V 20559 S116T,R37E 25429
Y15Q,R88Y 10820 N95Q,S116K 15690 L84E,A48G 20560 S116T,R37A 25430
Y15Q,R88D 10821 N95Q,S116H 15691 L84E,E53A 20561 S116T,S38A 25431
Y15Q,L89A 10822 N95Q,S116Q 15692 L84E,Y55A 20562 S116T,D39A 25432
Y15Q,E90K 10823 N95Q,C117K 15693 L84E,Y55F 20563 S116T,Q40A 25433
Y15Q,E91A 10824 N95Q,C117R 15694 L84E,K57A 20564 S116T,Q43A 25434
Y15Q,N92A 10825 N95Q,C117S 15695 L84E,K57E 20565 S116T,Q43E 25435
Y15Q,Y94V 10826 N95Q,C117Q 15696 L84E,E60A 20566 S116T,Q43K 25436
Y15Q,Y94A 10827 N95Q,C117T 15697 L84E,E60K 20567 S116T,S47A 25437
Y15Q,N95A 10828 N95Q,C117P 15698 L84E,T61A 20568 S116T,S47V 25438
Y15Q,N95V 10829 N95Q,C117A 15699 L84E,Q63A 20569 S116T,A48G 25439
Y15Q,T96A 10830 N95Q,C117I 15700 L84E,Y64A 20570 S116T,E53A 25440
Y15Q,K100E 10831 N95Q,K118W 15701 L84E,D68A 20571 S116T,Y55A 25441
Y15Q,K101E 10832 N95Q,K118H 15702 L84E,T78A 20572 S116T,Y55F 25442
Y15Q,H102A 10833 N95Q,K118R 15703 L84E,P79A 20573 S116T,K57A 25443
Y15Q,K105A 10834 N95Q,K118S 15704 L84E,E81K 20574 S116T,K57E 25444
Y15Q,K105E 10835 N95Q,K118M 15705 L84E,E87K 20575 S116T,E60A 25445
Y15Q,K113A 10836 N95Q,K118L 15706 L84E,E87Q 20576 S116T,E60K 25446
Y15Q,S116W 10837 N95Q,P121A 15707 L84E,E87V 20577 S116T,T61A 25447
Y15Q,S116Y 10838 N95Q,R122A 15708 L84E,E87D 20578 S116T,Q63A 25448
Y15Q,S116F 10839 N95Q,R122E 15709 L84E,R88L 20579 S116T,Y64A 25449
Y15Q,S116G 10840 N95Q,H124A 15710 L84E,R88Y 20580 S116T,D68A 25450 Y15Q,S116N 10841 N95Q,Q127A 15711 L84E,R88D 20581 S116T,T78A 25451
Y15Q,S116K 10842 N95Q,Q127K 15712 L84E,L89A 20582 S116T,P79A 25452
Y15Q,S116H 10843 N95Q,K128E 15713 L84E,E90K 20583 S116T,E81K 25453
Y15Q,S116Q 10844 N95Q,L133A 15714 L84E,E91A 20584 S116T,L84A 25454
Y15Q,C117K 10845 N95Q,L133V 15715 L84E,N92A 20585 S116T,E87K 25455
Y15Q,C117R 10846 N95R,K10E 15716 L84E,Y94V 20586 S116T,E87Q 25456
Y15Q,C117S 10847 N95R,Y15A 15717 L84E,Y94A 20587 S116T,E87V 25457
Y15Q,C117Q 10848 N95R,Y15F 15718 L84E,N95A 20588 S116T,E87D 25458
Y15Q,C117T 10849 N95R,S17A 15719 L84E,N95V 20589 S116T,R88L 25459
Y15Q,C117P 10850 N95R,S17K 15720 L84E,T96A 20590 S116T,R88Y 25460
Y15Q,C117A 10851 N95R,N18R 15721 L84E,K100E 20591 S116T,R88D 25461
Y15Q,C117I 10852 N95R,H21A 15722 L84E,K101E 20592 S116T,L89A 25462
Y15Q,K118
10853 N95R,L26A 15723 L84E,H102A 20593 S116T,E90K 25463 W
Y15Q,K118H 10854 N95R,P27A 15724 L84E,K105A 20594 S116T,E91A 25464
Y15Q,K118R 10855 N95R,D28K 15725 L84E,K105E 20595 S116T,N92A 25465
Y15Q,K118S 10856 N95R,T30A 15726 L84E,K113A 20596 S116T,Y94V 25466
Y15Q,K118
10857 N95R,T34A 15727 L84E,S116W 20597 S116T,Y94A 25467 M
Y15Q,K118L 10858 N95R,R35A 15728 L84E,S116Y 20598 S116T,N95A 25468
Y15Q,P121A 10859 N95R,R35E 15729 L84E,S116F 20599 S116T,N95V 25469
Y15Q,R122A 10860 N95R,R35F 15730 L84E,S116G 20600 S116T,T96A 25470
Y15Q,R122E 10861 N95R,R35G 15731 L84E,S116N 20601 S116T,K100E 25471
Y15Q,H124A 10862 N95R,R35I 15732 L84E,S116K 20602 S116T,K101E 25472
Y15Q,Q127A 10863 N95R,R35L 15733 L84E,S116H 20603 S116T,H102A 25473
Y15Q,Q127K 10864 N95R,R35M 15734 L84E,S116Q 20604 S116T,K105A 25474
Y15Q,K128E 10865 N95R,R35P 15735 L84E,C117K 20605 S116T,K105E 25475
Y15Q,L133A 10866 N95R,R35S 15736 L84E,C117R 20606 S116T,K113A 25476
Y15Q,L133V 10867 N95R,R35V 15737 L84E,C117S 20607 S116T,C117K 25477
Y15R,K10E 10868 N95R,D36A 15738 L84E,C117Q 20608 S116T,C117R 25478
Y15R,S17A 10869 N95R,D36K 15739 L84E,C117T 20609 S116T,C117S 25479
Y15R,S17K 10870 N95R,R37E 15740 L84E,C117P 20610 S116T,C117Q 25480
Y15R,N18R 10871 N95R,R37A 15741 L84E,C117A 20611 S116T,C117T 25481
Y15R,H21A 10872 N95R,S38A 15742 L84E,C117I 20612 S116T,C117P 25482
L84E,K118
Y15R,L26A 10873 N95R,D39A 15743 20613 S116T,C117A 25483
W
Y15R,P27A 10874 N95R,Q40A 15744 L84E,K118H 20614 S116T,C117I 25484
S116T,K118
Y15R,D28K 10875 N95R,Q43A 15745 L84E,K118R 20615 25485
W
Y15R,T30A 10876 N95R,Q43E 15746 L84E,K118S 20616 S116T,K118H 25486
Y15R,T34A 10877 N95R,Q43K 15747 L84E,K118M 20617 S116T,K118R 25487
Y15R,R35A 10878 N95R,S47A 15748 L84E,K118L 20618 S116T,K118S 25488
Y15R,R35E 10879 N95R,S47V 15749 L84E,P121A 20619 S116T,K118M 25489
Y15R,R35F 10880 N95R,A48G 15750 L84E,R122A 20620 S116T,K118L 25490
Y15R,R35G 10881 N95R,E53A 15751 L84E,R122E 20621 S116T,P121A 25491
Y15R,R35I 10882 N95R,Y55A 15752 L84E,H124A 20622 S116T,R122A 25492
Y15R,R35L 10883 N95R,Y55F 15753 L84E,Q127A 20623 S116T,R122E 25493
Y15R,R35M 10884 N95R,K57A 15754 L84E,Q127K 20624 S116T,H124A 25494 Y15R,R35P 10885 N95R,K57E 15755 L84E,K128E 20625 S116T,Q127A 25495
Y15R,R35S 10886 N95R,E60A 15756 L84E,L133A 20626 S116T,Q127K 25496
Y15R,R35V 10887 N95R,E60K 15757 L84E,L133V 20627 S116T,K128E 25497
Y15R,D36A 10888 N95R,T61A 15758 L84F,K10E 20628 S116T,L133A 25498
Y15R,D36K 10889 N95R,Q63A 15759 L84F,Y15A 20629 S116T,L133V 25499
Y15R,R37E 10890 N95R,Y64A 15760 L84F,Y15F 20630 S116V,K10E 25500
Y15R,R37A 10891 N95R,D68A 15761 L84F,S17A 20631 S116V,Y15A 25501
Y15R,S38A 10892 N95R,T78A 15762 L84F,S17K 20632 S116V,Y15F 25502
Y15R,D39A 10893 N95R,P79A 15763 L84F,N18R 20633 S116V,S17A 25503
Y15R,Q40A 10894 N95R,E81K 15764 L84F,H21A 20634 S116V,S17K 25504
Y15R,Q43A 10895 N95R,L84A 15765 L84F,L26A 20635 S116V,N18R 25505
Y15R,Q43E 10896 N95R,E87K 15766 L84F,P27A 20636 S116V,H21A 25506
Y15R,Q43K 10897 N95R,E87Q 15767 L84F,D28K 20637 S116V,L26A 25507
Y15R,S47A 10898 N95R,E87V 15768 L84F,T30A 20638 S116V,P27A 25508
Y15R,S47V 10899 N95R,E87D 15769 L84F,T34A 20639 S116V,D28K 25509
Y15R,A48G 10900 N95R,R88L 15770 L84F,R35A 20640 S116V,T30A 25510
Y15R,E53A 10901 N95R,R88Y 15771 L84F,R35E 20641 S116V,T34A 25511
Y15R,Y55A 10902 N95R,R88D 15772 L84F,R35F 20642 S116V,R35A 25512
Y15R,Y55F 10903 N95R,L89A 15773 L84F,R35G 20643 S116V,R35E 25513
Y15R,K57A 10904 N95R,E90K 15774 L84F,R35I 20644 S116V,R35F 25514
Y15R,K57E 10905 N95R,E91A 15775 L84F,R35L 20645 S116V,R35G 25515
Y15R,E60A 10906 N95R,N92A 15776 L84F,R35M 20646 S116V,R35I 25516
Y15R,E60K 10907 N95R,Y94V 15777 L84F,R35P 20647 S116V,R35L 25517
Y15R,T61A 10908 N95R,Y94A 15778 L84F,R35S 20648 S116V,R35M 25518
Y15R,Q63A 10909 N95R,T96A 15779 L84F,R35V 20649 S116V,R35P 25519
Y15R,Y64A 10910 N95R,K100E 15780 L84F,D36A 20650 S116V,R35S 25520
Y15R,D68A 10911 N95R,K101E 15781 L84F,D36K 20651 S116V,R35V 25521
Y15R,T78A 10912 N95R,H102A 15782 L84F,R37E 20652 S116V,D36A 25522
Y15R,P79A 10913 N95R,K105A 15783 L84F,R37A 20653 S116V,D36K 25523
Y15R,E81K 10914 N95R,K105E 15784 L84F,S38A 20654 S116V,R37E 25524
Y15R,L84A 10915 N95R,K113A 15785 L84F,D39A 20655 S116V,R37A 25525
Y15R,E87K 10916 N95R,S116W 15786 L84F,Q40A 20656 S116V,S38A 25526
Y15R,E87Q 10917 N95R,S116Y 15787 L84F,Q43A 20657 S116V,D39A 25527
Y15R,E87V 10918 N95R,S116F 15788 L84F,Q43E 20658 S116V,Q40A 25528
Y15R,E87D 10919 N95R,S116G 15789 L84F,Q43K 20659 S116V,Q43A 25529
Y15R,R88L 10920 N95R,S116N 15790 L84F,S47A 20660 S116V,Q43E 25530
Y15R,R88Y 10921 N95R,S116K 15791 L84F,S47V 20661 S116V,Q43K 25531
Y15R,R88D 10922 N95R,S116H 15792 L84F,A48G 20662 S116V,S47A 25532
Y15R,L89A 10923 N95R,S116Q 15793 L84F,E53A 20663 S116V,S47V 25533
Y15R,E90K 10924 N95R,C117K 15794 L84F,Y55A 20664 S116V,A48G 25534
Y15R,E91A 10925 N95R,C117R 15795 L84F,Y55F 20665 S116V,E53A 25535
Y15R,N92A 10926 N95R,C117S 15796 L84F,K57A 20666 S116V,Y55A 25536
Y15R,Y94V 10927 N95R,C117Q 15797 L84F,K57E 20667 S116V,Y55F 25537
Y15R,Y94A 10928 N95R,C117T 15798 L84F,E60A 20668 S116V,K57A 25538
Y15R,N95A 10929 N95R,C117P 15799 L84F,E60K 20669 S116V,K57E 25539
Y15R,N95V 10930 N95R,C117A 15800 L84F,T61A 20670 S116V,E60A 25540 Y15R,T96A 10931 N95R,C117I 15801 L84F,Q63A 20671 S116V,E60K 25541
Y15R,K100E 10932 N95R,K118W 15802 L84F,Y64A 20672 S116V,T61A 25542
Y15R,K101E 10933 N95R,K118H 15803 L84F,D68A 20673 S116V,Q63A 25543
Y15R,H102A 10934 N95R,K118R 15804 L84F,T78A 20674 S116V,Y64A 25544
Y15R,K105A 10935 N95R,K118S 15805 L84F,P79A 20675 S116V,D68A 25545
Y15R,K105E 10936 N95R,K118M 15806 L84F,E81K 20676 S116V,T78A 25546
Y15R,K113A 10937 N95R,K118L 15807 L84F,E87K 20677 S116V,P79A 25547
Y15R,S116W 10938 N95R,P121A 15808 L84F,E87Q 20678 S116V,E81K 25548
Y15R,S116Y 10939 N95R,R122A 15809 L84F,E87V 20679 S116V,L84A 25549
Y15R,S116F 10940 N95R,R122E 15810 L84F,E87D 20680 S116V,E87K 25550
Y15R,S116G 10941 N95R,H124A 15811 L84F,R88L 20681 S116V,E87Q 25551
Y15R,S116N 10942 N95R,Q127A 15812 L84F,R88Y 20682 S116V,E87V 25552
Y15R,S116K 10943 N95R,Q127K 15813 L84F,R88D 20683 S116V,E87D 25553
Y15R,S116H 10944 N95R,K128E 15814 L84F,L89A 20684 S116V,R88L 25554
Y15R,S116Q 10945 N95R,L133A 15815 L84F,E90K 20685 S116V,R88Y 25555
Y15R,C117K 10946 N95R,L133V 15816 L84F,E91A 20686 S116V,R88D 25556
Y15R,C117R 10947 N95S,K10E 15817 L84F,N92A 20687 S116V,L89A 25557
Y15R,C117S 10948 N95S,Y15A 15818 L84F,Y94V 20688 S116V,E90K 25558
Y15R,C117Q 10949 N95S,Y15F 15819 L84F,Y94A 20689 S116V,E91A 25559
Y15R,C117T 10950 N95S,S17A 15820 L84F,N95A 20690 S116V,N92A 25560
Y15R,C117P 10951 N95S,S17K 15821 L84F,N95V 20691 S116V,Y94V 25561
Y15R,C117A 10952 N95S,N18R 15822 L84F,T96A 20692 S116V,Y94A 25562
Y15R,C117I 10953 N95S,H21A 15823 L84F,K100E 20693 S116V,N95A 25563
Y15R,K118
10954 N95S,L26A 15824 L84F,K101E 20694 S116V,N95V 25564 W
Y15R,K118H 10955 N95S,P27A 15825 L84F,H102A 20695 S116V,T96A 25565
Y15R,K118R 10956 N95S,D28K 15826 L84F,K105A 20696 S116V,K100E 25566
Y15R,K118S 10957 N95S,T30A 15827 L84F,K105E 20697 S116V,K101E 25567
Y15R,K118
10958 N95S,T34A 15828 L84F,K113A 20698 S116V,H102A 25568 M
Y15R,K118L 10959 N95S,R35A 15829 L84F,S116W 20699 S116V,K105A 25569
Y15R,P121A 10960 N95S,R35E 15830 L84F,S116Y 20700 S116V,K105E 25570
Y15R,R122A 10961 N95S,R35F 15831 L84F,S116F 20701 S116V,K113A 25571
Y15R,R122E 10962 N95S,R35G 15832 L84F,S116G 20702 S116V,C117K 25572
Y15R,H124A 10963 N95S,R35I 15833 L84F,S116N 20703 S116V,C117R 25573
Y15R,Q127A 10964 N95S,R35L 15834 L84F,S116K 20704 S116V,C117S 25574
Y15R,Q127K 10965 N95S,R35M 15835 L84F,S116H 20705 S116V,C117Q 25575
Y15R,K128E 10966 N95S,R35P 15836 L84F,S116Q 20706 S116V,C117T 25576
Y15R,L133A 10967 N95S,R35S 15837 L84F,C117K 20707 S116V,C117P 25577
Y15R,L133V 10968 N95S,R35V 15838 L84F,C117R 20708 S116V,C117A 25578
Y15S,K10E 10969 N95S,D36A 15839 L84F,C117S 20709 S116V,C117I 25579
S116V,K118
Y15S,S17A 10970 N95S,D36K 15840 L84F,C117Q 20710 25580
W
Y15S,S17K 10971 N95S,R37E 15841 L84F,C117T 20711 S116V,K118H 25581
Y15S,N18R 10972 N95S,R37A 15842 L84F,C117P 20712 S116V,K118R 25582
Y15S,H21A 10973 N95S,S38A 15843 L84F,C117A 20713 S116V,K118S 25583
S116V,K118
Y15S,L26A 10974 N95S,D39A 15844 L84F,C117I 20714 25584
M Y15S,P27A 10975 N95S,Q40A 15845 L84F,K118W 20715 S116V,K118L 25585
Y15S,D28K 10976 N95S,Q43A 15846 L84F,K118H 20716 S116V,P121A 25586
Y15S,T30A 10977 N95S,Q43E 15847 L84F,K118R 20717 S116V,R122A 25587
Y15S,T34A 10978 N95S,Q43K 15848 L84F,K118S 20718 S116V,R122E 25588
Y15S,R35A 10979 N95S,S47A 15849 L84F,K118M 20719 S116V,H124A 25589
Y15S,R35E 10980 N95S,S47V 15850 L84F,K118L 20720 S116V,Q127A 25590
Y15S,R35F 10981 N95S,A48G 15851 L84F,P121A 20721 S116V,Q127K 25591
Y15S,R35G 10982 N95S,E53A 15852 L84F,R122A 20722 S116V,K128E 25592
Y15S,R35I 10983 N95S,Y55A 15853 L84F,R122E 20723 S116V,L133A 25593
Y15S,R35L 10984 N95S,Y55F 15854 L84F,H124A 20724 S116V,L133V 25594
Y15S,R35M 10985 N95S,K57A 15855 L84F,Q127A 20725
Y15S,R35P 10986 N95S,K57E 15856 L84F,Q127K 20726
Table 12.
Figure imgf000278_0001
Figure imgf000279_0001
Figure imgf000280_0001
ΔΝΤ9 H21A ΔΝΤ9
E87V S47V 25656 D36A K10E 25749 25842 25939
R35E R122T R35E
ΔΝΤ9
ΔΝΤ9 L26A
R88L S47V 25657 L84A K10E 25750 25843 R122V 25940
R35E
R35E
ΔΝΤ9
ΔΝΤ9 P27A
Y94V S47V 25658 E87V K10E 25751 25844 R122W 25941
R35E
R35E
ΔΝΤ9
ΔΝΤ9 D39A
N95A S47V 25659 R88L K10E 25752 25845 R122Y 25942
R35E
R35E
ΔΝΤ9
ΔΝΤ9 H41A
K101E S47V 25660 Y94V K10E 25753 25846 K112E 25943
R35E
R35E
ΔΝΤ9
S116W ΔΝΤ9 Q43A
25661 N95A K10E 25754 25847 R119A 25944 S47V R35E
R35E
ΔΝΤ9
K118W ΔΝΤ9 Q45A
25662 K101E K10E 25755 25848 H124A 25945 S47V R35E
R35E
ΔΝΤ9 S47A ΔΝΤ9 Q40A
Y15A E49A 25663 S116W K10E 25756 25849 25946
R35E R35E
K118W ΔΝΤ9 A48G ΔΝΤ9 S38A
D28A E49A 25664 25757 25850 25947
K10E R35E R35E
ΔΝΤ9 E49A ΔΝΤ9 E60A
R35A E49A 25665 R35A L84A 25758 25851 25948
R35E R35E
ΔΝΤ9 E49K WT L3A
D36A E49A 25666 R35A R88D 25759 25852 25954
R35E R35E
ΔΝΤ9 S50A WT P4A
L84A E49A 25667 R35A Q43E 25760 25853 25955
R35E R35E
R35A ΔΝΤ9 V51A ΔΝΤ9 Q63A
E87V E49A 25668 25761 25854 25956
K118W R35E R35E
ΔΝΤ9
ΔΝΤ9 E53A
R88L E49A 25669 L84A R88D 25762 25855 K118E 25957
R35E
R35E
ΔΝΤ9 Y55A ΔΝΤ9 T69A
Y94V E49A 25670 L84A Q43E 25763 25856 25958
R35E R35E
ΔΝΤ9
L84A ΔΝΤ9 K57A
N95A E49A 25671 25764 25857 H102A 25959
K118W R35E
R35E
ΔΝΤ9 K57E ΔΝΤ9 T30A
K101E E49A 25672 R88D Q43E 25765 25858 25960
R35E R35E
S116W R88D ΔΝΤ9 S58A ΔΝΤ9 D68A
25673 25766 25859 25961 E49A K118W R35E R35E
K118W Q43E ΔΝΤ9 T59A ΔΝΤ9 E81K
25674 25767 25860 25962 E49A K118W R35E R35E
ΔΝΤ9 E60K ΔΝΤ9 D28K
Y15A E53A 25675 L84A L133A 25768 25861 25963
R35E R35E
ΔΝΤ9 R88D ΔΝΤ9 T61A ΔΝΤ9 D36K
D28A E53A 25676 25769 25862 25964
L133A R35E R35E
ΔΝΤ9 Q43E ΔΝΤ9 D70A ΔΝΤ9 D32K
R35A E53A 25677 25770 25863 25965
L133A R35E R35E
ΔΝΤ9 ΔΝΤ9
ΔΝΤ9 L72A
D36A E53A 25678 K118W 25771 25864 K100E 25966
R35E
L133A R35E
ΔΝΤ9 N95A ΔΝΤ9 S76A ΔΝΤ9 R37A
L84A E53A 25679 25772 25865 25967
L133A R35E R35E
ΔΝΤ9 T34A ΔΝΤ9 Q77A ΔΝΤ9
E87V E53 A 25680 25773 25866 25968
L133A R35E K118A R35E
ΔΝΤ9 R37A ΔΝΤ9 T78A ΔΝΤ9 T34A
R88L E53A 25681 25774 25867 25969
L133A R35E R35E
ΔΝΤ9 P79A ΔΝΤ9 D36A
Y94V E53 A 25682 ΔΝΤ9 N92T 25775 25868 25970
R35E R35E
ΔΝΤ9 T96A ΔΝΤ9 N80A
N95A E53A 25683 25776 25869 ΔΝΤ9 R88L 25971
N92A R35E
ΔΝΤ9 T96A ΔΝΤ9 E81A
K101E E53A 25684 25777 25870 ΔΝΤ9 R88D 25972
K57E R35E
S116W ΔΝΤ9 ΔΝΤ9 E82A
25685 25778 25871 ΔΝΤ9 Q43E 25973
E53A E91K/ 57E R35E
K118W ΔΝΤ9 N92A ΔΝΤ9 E82K
25686 25779 25872 ΔΝΤ9 Y15A 25974 E53A K57E R35E
ΔΝΤ9 ΔΝΤ9 L84A
Y15A E60A 25687 25780 25873
A48G/ 57E R35E
Example 2
[0368] Molecular Biology and Cloning: Mutagenesis primers were ordered from Integrated DNA Technologies and Eton Biosciences. Constructs were created using established methods involving circular polymerase extension cloning or Gibson assembly.
[0369] Small Scale Expression & Purification: Wild-type FGF1 and variants were expressed from pET28(+)/BL21(DE3) E. coli expression system. Protein expression from a 0.05 L culture was induced with 0.5 mM IPTG overnight at 20°C. Cell pellets were lysed with 20 mM Tris buffer pH 7.4, 0.5 M NaCl, 0.1 mg/ml PMSF, 1 mM EDTA, 0.10 mg/mL lysozyme, and supplemented with
Benzonase® Nuclease. Variants were purified by heparin sepharose affinity and eluted with 1.5 M NaCl. Eluted samples were evaluated for purity by analytical size exclusion chromatography using a TSKgel G2000SWxl from Tosoh Bioscience and SDS-PAGE. Protein concentration was determined using absorbance at 280 nm and each variant's extinction coefficient.
[0370] Protein Expression and Purification: Recombinant wild-type FGF1 and variants were expressed from pET28(+)/BL21(DE3) E. coli expression system. Protein expression from a 0.5L culture was induced by 0.5 mM isopropyl- -D-thio-galactoside (IPTG) at OD600 between 0.6 - 1.0, with simultaneous reduction of incubation temperature from 37 °C to 20 °C, followed by overnight incubation. Expression cultures were harvested and lysed in 20mM Tris buffer pH 7.4 with 0.5 M NaCl, 0.1 mg/mL phenylmethylsulfonyl fluoride (PMSF), ImM ethylenediaminetetraacetic acid (EDTA), 0.25mg/mL lysozyme (Sigma) and supplemented with Benzonase® Nuclease (Millipore). The expressed protein variants were purified by sequential chromatography on heparin sepharose resin (GE Healthcare Life Sciences) and SP sepharose resin (GE Healthcare Life Sciences). The purified proteins were treated with endotoxin removal resin (Thermo Scientific), and the endotoxin levels of treated proteins were determined by ToxinSensor Chrmongenic LAL Endotoxin Assay kit (Genscript). The protein variants with acceptable endotoxin level (< 0.5 EU/mL) were evaluated by analytical size exclusion chromatography (TSKgel G2000SWxl - Tosoh Bioscience) to confirm that the protein samples were homogeneous and folded. Protein purity was also evaluated by gel densitometry of Coomassie Brilliant Blue stained SDS-PAGE. The extinction coefficient calculated from ProtParam program based on each variant amino acid sequence was utilized for concentration determination.
[0371] Determination of Tm by Differential Scanning Fluorimetry: Melting temperatures of lead compounds were determined essentially as described (Simeonov, Expert Opin. Drug Discovery (2013) vol. 8 no. 9 1071-1082; Boivin et al, Protein Expr. Purif. (2013) vol. 91 no. 2 192-206), with the following adjustments. Proteins were diluted to 50 μg/ml in buffer containing 5 x SYPRO Orange Protein Gel Stain (Sigma-Aldrich) with TBS 130 (Tris-HCl pH 7.5, 130 mM NaCl), TBS530 (Tris- HC1 pH 7.5, 530 mM NaCl), PBS 130 (Phosphate-Buffered Saline pH 7.4, 130 mM NaCl), PBS530 (Phosphate-Buffered Saline pH 7.4, 130 mM NaCl), or Buffer A (20 mM Histidine pH 7.2, 75 mM Potassium Sulfate, 75 mM NaCl). Proteins in buffered solution (20 μΐ final volume) were loaded into a 384-well Microamp optical plate (Thermo Fisher Scientific) and run on an ABI7900 qPCR thermocycler. The temperature of the plate was increased from 25 °C to 95 °C at 1 °C/min ramp rate and raw data was collected using the ROX detection filter (no background filter applied). The raw data file was exported as a multicomponent text data file and read by the DSF software DMAN (Wang et al, Bioinformatics (2012) vol. 28 no. 3 439-440). Melting temperatures were determined by applying the maximal first-derivative method using the DMAN software.
Example 3
[0372] Cell Proliferation Assays: Cell proliferation assays were performed in NIH3T3 cells or AML12 cells purchased from ATCC. Cells were maintained in either DMEM media containing 10% new born calf serum (NBCS) for NIH3T3 cells or DMEM/F12 media containing 10% fetal bovine serum (FBS) for AML12 cells. To assess FGF1 induced cell proliferation, cells were seeded at 10000-15000 cells per well in white-walled clear bottom 96 well cell culture treated assay plates coated with poly-L-lysine. At 16-18hr after plating, cells were wash with PBS and starvation media (DMEM with 0.5% NBCS for NIH3T3 or serum free DMEM/F12 media for AML12 cells) was added. After 24 hours of starvation, FGF 1 variants were added and 24hr later cells were labeled with BrdU for 3 hrs. After 3 hr BrdU labeling, cell proliferation was assessed using the Roche Colormetric BrdU Quantification Assay according to kit manufacturer's instructions.
[0373] Table 13 illustrates the mitogenicity of the FGF1 polypeptides tested using the NIH3T3 cell proliferation assay. The mitogenicity ranking is based on 3 dose levels. The ranking is as follows: 1 is inactive at all doses tested; 2 is inactive at 2 out of 3 doses tested; 3 is <80% of WT-FGFl; 4 equals to WT-FGFl; and 5 is more mitogenic than WT-FGFl .
Table 13. NIH3T3 Cell Proliferation Assay FGF1 Relative FGF1 Relative FGF1 Relative FGF1 Relative ID Mitogenicity ID Mitogenicity ID Mitogenicity ID Mitogenicity
MP168 4 MP232 4 MP291 3 MP624 3
MP169 4 MP233 4 MP294 4 MP625 2
MP170 4 MP234 4 MP296 4 MP626 5
MP171 4 MP236 4 MP297 4 MP627 2
MP172 4 MP237 4 MP298 4 MP628 2
MP173 4 MP238 3 MP299 4 MP629 1
MP174 4 MP239 3 MP300 4 MP630 2
MP175 3 MP240 4 MP301 4 MP631 1
MP176 5 MP241 4 MP577 4 MP632 2
MP177 4 MP242 3 MP578 5 MP633 5
MP178 4 MP243 4 MP579 3 MP634 5
MP181 4 MP244 4 MP580 3 MP635 5
MP182 5 MP245 2 MP581 5 MP636 1
MP183 3 MP246 4 MP582 4 MP638 2
MP185 5 MP247 4 MP583 5 MP639 2
MP187 2 MP248 2 MP584 4 MP640 1
MP191 3 MP249 4 MP585 4 MP641 3
MP192 3 MP250 4 MP586 3 MP643 3
MP194 2 MP251 3 MP588 5 MP644 1
MP195 3 MP252 4 MP589 4 MP645 1
MP198 3 MP253 3 MP590 4 MP646 2
MP199 2 MP254 3 MP591 4 MP647 2
MP200 2 MP256 3 MP592 2 MP648 1
MP201 2 MP257 4 MP593 3 MP649 2
MP202 2 MP258 2 MP595 4 MP650 2
MP203 3 MP259 2 MP597 3 MP653 2
MP204 2 MP260 3 MP598 4 MP654 2
MP205 3 MP261 4 MP599 5 MP655 2
MP206 3 MP263 3 MP600 5 MP656 3
MP208 5 MP264 4 MP602 4 MP657 5
MP210 3 MP265 2 MP603 2 MP660 5
MP211 4 MP266 3 MP604 2 MP661 3
MP213 3 MP267 4 MP606 4 MP662 4
MP214 5 MP268 4 MP607 2 MP665 4
MP215 4 MP269 2 MP608 4 MP690 1
MP216 4 MP270 3 MP609 2 MP692 1
MP217 4 MP271 4 MP610 3 MP693 5
MP218 4 MP274 4 MP611 3 MP695 2
MP220 3 MP276 3 MP612 4 MP697 2
MP221 3 MP279 4 MP613 3 MP698 1
MP222 3 MP282 4 MP614 3 MP699 2
MP223 4 MP283 4 MP615 3 MP872 2
MP224 4 MP284 3 MP616 3 MP873 2 MP225 5 MP285 5 MP617 4 MP875 2
MP226 5 MP286 5 MP618 3 MP879 2
MP227 3 MP287 5 MP619 4 MP882 2
MP228 3 MP288 4 MP620 3 MP883 2
MP229 5 MP289 3 MP622 4 MP884 1
MP230 3 MP290 4 MP623 5
[0374] Table 14 shows the mitogenicity of the FGFl polypeptides tested using the AML12 cell proliferation assay. The mitogenicity activity or ranking is based on 3 dose levels. Ranking or level of activity is denoted by (*) to (****) in which (*) indicates the lowest mitogenicity and (****) indicates the highest mitogenicity. (****) also indicates wild-type activity level. The Wild-type (WT) FGFl polypeptide is the mature form, residue 1-140.
Table 14.
Figure imgf000285_0001
Figure imgf000286_0001
Figure imgf000287_0001
Figure imgf000288_0001
[0375] FGF l polypeptide comprising a point mutation at, e.g., F22, P27, D28, R35, D36, R37, E53, K57, T61, D68, Y74, E81, E87, N92, Y94, T96, K100, KlO l, H102, F108, Nl 14, Kl 18, Rl 19, Q 127, K128, and L135; and an N-terminal truncation of the first 9 residues showed lower mitogenic activity relative to the wild-type (WT) FGF l mature form. Truncated FGFl polypeptides with a first mutation at residue R35 or R37 and a second mutation at Q43, A48, Y74, E91, N92, H93, 198, K101, R122, or VI 37 exhibited the lowest mitogenic activity relative to other tested FGF l mutants and the WT FGF l polypeptide.
Example 4
[0376] FGFRlc Gal4-ELK1 Activity: Fibroblast growth factor receptor lc (FGFRlc), also known as FGFR1 isoform 2, is one of the receptor binding partners of FGFl . Exemplary truncated FGFl polypeptides comprising point mutations described herein (with an N-terminal deletion of the first 9 residues) were screened for FGFRlc binding interaction in a FGFRlc Gal4-ELK1 reporter cell assay and further screened for mitogenic activity in AML12 cells.
[0377] Rat L6 myoblast cell line was maintained in DMEM media with 10% FBS. Cells were plated in at 10,000 cells/well in clear bottom 96 well plates. The day after plating, the cells were transfected with DNA expression plasmids encoding FGFRlc, Gal4Elkl, PFR luciferase reporter and YFP expression plasmid using Xtremgen transfection reagent. The following day transfection media was removed and cells were incubated with FGF 1 protein variants in the presence or absence of lug/ml Heparin in DMEM serum free media for 6 hours. Cells were then lysed and luciferase and YFP signals were measured with the YFP used to normalize the luciferase values.
[0378] Table 15 illustrates the Gal4-ELK1 activity of exemplary FGF1 polypeptides described herein.
Table 15
Figure imgf000289_0001
ΔΝΤ9
ΔΝΤ9 E90A
25880 MP2814 +++ 25902 R119E MP2845 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 S50A
25853 MP2778 +++ 25884 N92A MP2818 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 R122A
25923 MP2847 +++ 25898 K112A MP2836 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 P79A
25868 MP2801 +++ 25881 E90K MP2815 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9
25925 MP2862 +++ 25843 L26A MP2753 + D140A R35E
R35E
ΔΝΤ9
ΔΝΤ9 E91A
25882 MP2816 +++ 25940 R122V MP2877 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 E60A
25948 MP2787 +++ 25858 K57E MP2784 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 R122L
25933 MP2870 +++ 25895 K105E MP2832 +
R35E
R35E
ΔΝΤ9
WT P5A
25831 MP2735 +++ 25890 K100A MP2824 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 S58A
25859 MP2785 +++ 25873 L84A MP2807 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 E49A
25851 MP2776 +++ 25892 K101E MP2827 +
R35E
R35E
ΔΝΤ9
WT F1A
25829 MP2731 +++ 25862 T61A MP2789 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 T59A
25860 MP2786 +++ 25959 H102A MP2828 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 K10A
25836 MP2743 +++ 25903 P121A MP2846 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 R122Y
25942 MP2879 +++ 25960 T30A MP2757 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9
25906 MP2850 +++ 25961 D68A MP2792 + Y125A R35E
R35E
ΔΝΤ9
WT N2A
25830 MP2732 +++ 25887 N95A MP2821 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 Κ112Ε
25943 MP2837 +++ 25962 E81K MP2804 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 E82A
25871 MP2805 +++ 25856 Y55A MP2782 +
R35E
R35E
ΔΝΤ9 R122S ΔΝΤ9
25938 MP2875 +++ 25909 MP2853 +
R35E K128E R35E
ΔΝΤ9
ΔΝΤ9 Q77A
25866 MP2799 +++ 25839 Y15A MP2746 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 T78A
25867 MP2800 +++ 25896 N106A MP2833 +
R35E
R35E
ΔΝΤ9
WT G6A
25832 MP2736 +++ 25913 L135A MP2857 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 R122F
25928 MP2865 +++ 25845 D39A MP2768 +
R35E
R35E
ΔΝΤ9
WT K9A
25835 MP2739 +++ 25963 D28K MP2756 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 R119A
25944 MP2844 +++ 25872 E82K MP2806 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 R122K
25932 MP2869 +++ 25964 D36K MP2764 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 R122H
25930 MP2867 +++ 25965 D32K MP2759 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 E49K
25852 MP2777 +++ 25929 R122G MP2866 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 Q40A
25946 MP2770 +++ 25899 K113A MP2838 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 V51A
25854 MP2779 +++ 25841 N18A MP2748 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 P136A
25914 MP2858 +++ 25844 P27A MP2754 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 D70A
25863 MP2795 +++ 25907 Q127A MP2851 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 N80A
25869 MP2802 +++ 25886 Y94A MP2820 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 A48G
25850 MP2775 +++ 25931 R122I MP2868 +
R35E
R35E
ΔΝΤ9
WT L3A
25954 MP2733 +++ 25878 R88E MP2812 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 E81A
25870 MP2803 ++ 25894 K105A MP2831 +
R35E
R35E
ΔΝΤ9
WT P4A
25955 MP2734 ++ 25888 T96A MP2822 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 S116A
25901 MP2841 ++ 25966 K100E MP2825 +
R35E
R35E
25893 ΔΝΤ9 E104K MP2830 ++ 25897 ΔΝΤ9 MP2835 + R35E F108A
R35E
ΔΝΤ9
ΔΝΤ9 P134A
25912 MP2856 ++ 25967 R37A MP2765 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 I98A
25889 MP2823 ++ 25968 K118A MP2842 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 L72A
25864 MP2796 ++ 25969 T34A MP2760 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 S139A
25917 MP2861 ++ 25970 D36A MP2763 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 S47A
25849 MP2774 ++ 25846 H41A MP2771 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9
25915 MP2859 ++ 25874 L86A MP2808 + V137A R35E
R35E
ΔΝΤ9
ΔΝΤ9 S76A
25865 MP2798 ++ 25910 L131A MP2854 +
R35E
R35E
ΔΝΤ9
WT N7A
25833 MP2737 ++ 25876 E87K MP2810 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 Q45A
25848 MP2773 ++ 25842 H21A MP2749 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 H93A
25885 MP2819 ++ 25900 K113E MP2839 +
R35E
R35E
ΔΝΤ9
ΔΝΤ9 Q43A
25847 MP2772 ++ 25911 L133A MP2855 +
R35E
R35E
ΔΝΤ9 Q63A
25956 MP2790 ++
R35E
Example 5
[0379] Table 16 illustrates the Gal4-ELK1 activity and the mitogenic activity per residue. The FGFRlc Gal4-ELK1 activity ranking is denoted by (+) to (++++), in which (++++) denotes wild-type activity. The mitogenicity ranking is denoted by (*) to (****) in which (****) indicates wild-type activity.
Table 16
Figure imgf000292_0001
Figure imgf000293_0001
L133 +++ **
Residue numbering is based on the FGF1 mature form (residue 1-140)
[0380] Residues P27, R35, D36, R37, Y55, K57, T61, E81, E87, R88, N92, T96, K101, H102, and L133, categorized as Group 1 residues, exhibited near wild-type FGFRlc interaction and low mitogenicity. Residues L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, and G120, categorized as Group 2 residues, exhibited higher mitogenicity relative to the Group 1 residues and near wild-type FGFRlc interaction.
[0381] Group 1 and Group 2 residues are further mapped onto a crystal structure of FGFl-FGFRlc complex (PDB: 30JV.pdb). Fig. 1 and Fig. 2 show two different orientations of the FGFl-FGFRlc complex. Fig. 1A and Fig. 2A illustrate the positions of Group 1A, Group IB, and Group 1C residues relative to the FGFl-FGFRlc complex. Group 1C residues are proximal to the βΟ'-βΕ strands of FGFRlc and form a "smile" orientation on FGF1.
[0382] Fig. 3 and Fig. 4 illustrate the locations of the Group 1 and Group 2 residues relative to the FGFl-FGFRlc complex in two different orientations. Group 2 residues are dispersed around the Group 1C residues.
[0383] In some instances, an FGF 1 polypeptide comprising a combination of one or more Group 1 mutations and one or more Group 2 mutations exhibit near wild-type FGFRlc interaction and low mitogenicity. In some cases, an FGF1 polypeptide comprising a combination of a single Group 1 mutation and one or more Group 2 mutations exhibit near wild-type FGFRlc interaction and low mitogenicity.
[0384] Fig. 5A-Fig. 5D show exemplary residue candidates for mutation from Examples 3 and 4 (Fig. 5A and Fig. 5B) and their locations relative to Group 1 residues (Fig. 5C and Fig. 5D). Fig. 5B is a close-up of the position of the exemplary residues. Fig. 5D is a close-up of the positions of the exemplary residues and the Group 1 residues. The exemplary residues from Examples 3 and 4 include N7, S38, Q43, Q45, A48, S58, T78, P79, N80, E81, E82, E91, H93, 198, K101, P136, and V137 as shown in Fig. 5B; and N7, S17, S38, Q40, Q43, Q45, S47, E49, S50, V51, E53, S58, E60, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, 198, K101, E104, Kl 12, S I 16, R122, Y125, P136, V137, S138, S 139, and D140 as shown in Fig. 5D. In some instances, the exemplary residues from Examples 3 and 4 exhibit wild-type or near wild-type FGFRlc interaction and low mitogenicity. In some instances, the exemplary residues exhibit wild-type or near wild-type FGFRlc interaction relative to the Group 1 residues and low mitogenicity relative to the Group 1 residues. In some cases, a mutation at residue E81, N92, and/or K101 lead to wild-type or near wild-type FGFRlc interaction and/or low mitogenicity.
Example 6
[0385] Assessment of in vivo mitogenicity [0386] The adrenal cortex is the primary site of steroid synthesis, producing glucocorticoids under the control of the hypothalamic-pituitary-adrenal (HP A) axis and mineralocorticoids under the control of the renin-angiotensin system (RAS). Interaction of FGFl with FGFRs activates a cascading effect, leading to adrenocortical development and maintenance. In some instances, FGFl also acts as a mitogen to adrenocortical cells. Assessment of different adrenal tissues and hepatocytes was conducted to evaluate the mitogenicity of exemplary FGFl polypeptides.
[0387] Study day 1, osmotic minipumps (ALZET, Cupertino, CA) containing 5-bromo-2- deoxyuridine (20mg/ml in saline,) implanted subcutaneously into 9-week old, male, C57BL6 mice (n=4/group). Days 2-7 , daily subcutaneous injections of vehicle (Tris/Nacl buffer diluted 1 :5 in PBS), WT-FGF1 or FGFl mutants at (lmg/kg/day) . Day 8, 24 hrs after the last dose of test article, tissues collected and fixed in 10% neutral buffered formalin. Fixed tissues subsequently embedded in paraffin and processed for BrdU immunolabeling and quantitation.
[0388] Tables 17-20 and Figs. 6-9 illustrate the mitogenic activity of exemplary FGFl
polypeptides in zona glomerulosa (Table 17, Fig. 6), zona fasciculata/reticularis (Table 18, Fig. 7), medulla (Table 19, Fig. 8), and hepatocytes (Table 20, Fig. 9).
Table 17
Adrenal - Glomerulosa BrdU positive Nuclei
Treatment / Animal Total BrdU Positive % BrdU Positive
Group Dose # Cells Cells Cells
1 1 1414 220 15.5587
1 2 895 175 19.5531
Vehicle
1 3 1024 205 20.0195
1 4 1407 277 19.6873
MEAN 18.7047
STDEV 2.1064
2 5 2016 1293 64.1369
2 WT-FGF1 6 3200 2097 65.5313
2 lmg/kg 7 3215 2020 62.8305
2 8 2545 1640 64.4401
MEAN 64.2347
STDEV 1.1113
3 9 1483 653 44.0324
3 10 1527 724 47.4132
MP200 lmg/kg
3 11 1632 681 41.7279
3 12 1377 588 42.7015
MEAN 43.9688
STDEV 2.4830 13 1321 576 43.6033
14 840 278 33.0952
MP200 3mg/kg
15 1651 817 49.4852
16 1573 738 46.9167
MEAN 43.2751
STDEV 7.2010
17 1663 804 48.3464
18 1865 769 41.2332
MP200 lOmg/kg
19 2226 1077 48.3827
20 1451 653 45.0034
MEAN 45.7414
STDEV 3.3976
21 1516 436 28.7599
22 1607 548 34.1008
MP293 lmg/kg
23 1758 643 36.5757
24 1251 508 40.6075
MEAN 35.0110
STDEV 4.9556
25 1686 688 40.8066
26 738 476 64.4986
MP293 lOmg/kg
27 2298 1224 53.2637
28 1676 792 47.2554
MEAN 51.4561
STDEV 10.0736
29 2264 988 43.6396
30 1831 873 47.6789
MP697 lmg/kg
31 1823 782 42.8963
32 1829 970 53.0344
MEAN 46.8123
STDEV 4.6500
33 2190 1078 49.2237
34 1460 798 54.6575
MP697 lOmg/kg
35 1724 824 47.7958
36 1631 914 56.0392
MEAN 51.9291
STDEV 4.0307
37 1812 734 40.5077
38 1114 440 39.4973
MP 1834 lmg/kg
39 1904 798 41.9118
40 1589 643 40.4657
MEAN 40.5956
STDEV 0.9939 41 1651 869 52.6348
MP1834 42 1636 703 42.9707 lOmg/kg 43 2016 933 46.2798
44 2092 928 44.3595
MEAN 46.5612
STDEV 4.2703
45 1827 995 54.4609
46 2303 1053 45.723
MP2776 lmg/kg
47 1942 859 44.2327
48 2349 1291 54.9596
MEAN 49.8441
STDEV 5.6555
49 1997 906 45.3681
MP2776 50
lOmg/kg 51 1897 1094 57.67
52 1949 984 50.4874
MEAN 51.1752
STDEV 6.1797
53 1925 748 38.8571
54 2077 853 41.0688
MP2814 lmg/kg
55 1722 696 40.4181
56 2168 776 35.7934
MEAN 39.0344
STDEV 2.3515
57 1637 576 35.1863
MP2814 58 1873 694 37.0529 lOmg/kg 59 1405 665 47.331
60 2002 1020 50.9491
MEAN 42.6298
STDEV 7.6989
61 1634 710 43.4517
62 1318 530 40.2124
MP2847 lmg/kg
63 1624 713 43.9039
64 1282 350 27.3011
MEAN 38.7173
STDEV 7.7863
65 1655 691 41.7523
MP2847 66 2196 822 37.4317 lOmg/kg 67 1817 884 48.6516
68 1467 641 43.6946 MEAN 42.8826
STDEV 4.6523
18 69 1480 566 38.2432
18 70 1830 845 46.1749
MP2850 lmg/kg
18 71 1628 739 45.3931
18 72 1632 689 42.2181
MEAN 43.0073
STDEV 3.6076
19 73 2140 948 44.2991
19 MP2850 74 1592 746 46.8593
19 lOmg/kg 75 1995 909 45.5639
19 76 1968 953 48.4248
MEAN 46.2868
STDEV 1.7675
20 77 1481 635 42.8764
20 78 1825 618 33.863
MP2859 lmg/kg
20 79 1779 853 47.9483
20 80
MEAN 41.5626
STDEV 7.1340
21 81 2377 1299 54.6487
21 MP2859 82 1262 794 62.916
21 lOmg/kg 83 1467 660 44.9898
21 84 1125 625 55.5556
MEAN 54.5275
STDEV 7.3577
Table 18
Adrenal - Fasiculata/Reticularis BrdU
Treatment / Animal Total BrdU Positive % BrdU Positive
Group Dose # Cells Cells Cells
1 1 2145 40 1.8648
1 2 2400 134 5.58333
Vehicle
1 3 2404 136 5.65724
1 4 2367 88 3.71779
MEAN 4.2058
STDEV 1.8003
2 5 1666 75 4.5018
2 WT-FGF1 6 2795 273 9.76744
2 lmg/kg 7 3044 321 10.5453
2 8 1694 128 7.55608 MEAN 8.0927
STDEV 2.7082
9 2248 60 2.66904
10 2058 43 2.08941
MP200 lmg/kg
11 2210 35 1.58371
12 1442 50 3.46741
MEAN 2.4524
STDEV 0.8090
13 2231 45 2.01703
14 1559 29 1.86017
MP200 3mg/kg
15 2966 63 2.12407
16 2138 45 2.10477
MEAN 2.0265
STDEV 0.1203
17 2166 136 6.27886
18 2634 83 3.1511
MP200 lOmg/kg
19 2189 144 6.57835
20 1254 34 2.71132
MEAN 4.6799
STDEV 2.0309
21 1311 59 4.50038
22 1769 71 4.01357
MP293 lmg/kg
23 2056 82 3.98833
24 1237 69 5.57801
MEAN 4.5201
STDEV 0.7436
25 2234 45 2.01432
26 1152 53 4.60069
MP293 lOmg/kg
27 2480 104 4.19355
28 2120 52 2.45283
MEAN 3.3153
STDEV 1.2728
29 3671 41 1.11686
30 2947 51 1.73057
MP697 lmg/kg
31 2725 53 1.94495
32 2030 54 2.6601
MEAN 1.8631
STDEV 0.6367
33 2552 127 4.97649
MP697 lOmg/kg
34 1897 111 5.85134 35 1853 95 5.12682
36 1933 99 5.12157
MEAN 5.2691
STDEV 0.3944
37 2136 24 1.1236
38 1495 22 1.47157
MP 1834 lmg/kg
39 2018 28 1.38751
40 1529 35 2.28908
MEAN 1.5679
STDEV 0.5031
41 1808 89 4.92257
MP1834 42 2074 54 2.60366 lOmg/kg 43 2804 114 4.06562
44 2526 54 2.13777
MEAN 3.4324
STDEV 1.2890
45 2225 151 6.78652
46 2512 61 2.42834
MP2776 lmg/kg
47 2429 105 4.32277
48 2565 78 3.04094
MEAN 4.1446
STDEV 1.9300
49 1963 85 4.33011
MP2776 50
lOmg/kg 51 2327 57 2.44951
52 2398 95 3.96163
MEAN 3.5804
STDEV 0.9966
53 2948 44 1.49254
54 3305 100 3.02572
MP2814 lmg/kg
55 2608 35 1.34202
56 3301 39 1.18146
MEAN 1.7604
STDEV 0.8530
57 2301 28 1.21686
MP2814 58 2303 30 1.30265 lOmg/kg 59 2611 39 1.49368
60 2323 57 2.45372
MEAN 1.6167
STDEV 0.5699 16 61 2467 90 3.64816
16 62 2679 51 1.9037
MP2847 lmg/kg
16 63 2615 38 1.45315
16 64 2597 21 0.808625
MEAN 1.9534
STDEV 1.2159
17 65 2221 59 2.65646
17 MP2847 66 2966 29 0.977748
17 lOmg/kg 67 1764 42 2.38095
17 68 2458 20 0.81367
MEAN 1.7072
STDEV 0.9461
18 69 2211 44 1.99005
18 70 2675 89 3.3271
MP2850 lmg/kg
18 71 2132 30 1.40713
18 72 2617 28 1.06993
MEAN 1.9486
STDEV 0.9945
19 73 2475 66 2.66667
19 MP2850 74 1774 83 4.67869
19 lOmg/kg 75 3159 100 3.16556
19 76 2339 74 3.16375
MEAN 3.4187
STDEV 0.8722
20 77 1973 47 2.38216
20 78 2370 42 1.77215
MP2859 lmg/kg
20 79 2285 70 3.06346
20 80
MEAN 2.4059
STDEV 0.6460
21 81 2780 93 3.34532
21 MP2859 82 2147 67 3.12063
21 lOmg/kg 83 2206 87 3.94379
21 84 1923 83 4.31617
MEAN 3.6815
STDEV 0.5475
Table 19
Adrenal - Medulla BrdU positive Nuclei
Treatment / Animal Total BrdU Positive % BrdU Positive
Group Dose # Cells Cells Cells 1 319 6 1.88088
2 344 26 7.55814
Vehicle
3 327 10 3.0581
4 501 23 4.59082
MEAN 4.2720
STDEV 2.4557
5 444 166 37.3874
WT-FGF1 6 877 275 31.3569 lmg/kg 7 945 243 25.7143
8 436 221 50.6881
MEAN 36.2867
STDEV 10.7190
9 275 25 9.09091
10 395 65 16.4557
MP200 lmg/kg
11 191 23 12.0419
12 118 18 15.2542
MEAN 13.2107
STDEV 3.3189
13 195 24 12.3077
14 276 60 21.7391
MP200 3mg/kg
15 335 30 8.95522
16 465 80 17.2043
MEAN 15.0516
STDEV 5.5992
17 204 45 22.0588
18 556 137 24.6403
MP200 lOmg/kg
19 685 258 37.6642
20 95 37 38.9474
MEAN 30.8277
STDEV 8.7148
21 99 27 27.2727
22 323 132 40.8669
MP293 lmg/kg
23 270 75 27.7778
24 239 105 43.9331
MEAN 34.9626
STDEV 8.6811
25 570 373 65.4386
26 353 221 62.6062
MP293 lOmg/kg
27 545 319 58.5321
28 409 270 66.0147
MEAN 63.1479 STDEV 3.4188
29 720 74 10.2778
30 526 62 11.7871
MP697 lmg/kg
31 465 58 12.4731
32 525 124 23.619
MEAN 14.5393
STDEV 6.1222
33 948 172 18.1435
34 508 94 18.5039
MP697 lOmg/kg
35 460 135 29.3478
36 386 71 18.3938
MEAN 21.0973
STDEV 5.5024
37 594 27 4.54545
38 300 42 14
MP 1834 lmg/kg
39 621 29 4.66989
40 277 27 9.74729
MEAN 8.2407
STDEV 4.5404
41 216 60 27.7778
MP1834 42 684 156 22.807 lOmg/kg 43 820 148 18.0488
44 714 80 11.2045
MEAN 19.9595
STDEV 7.0601
45 610 190 31.1475
46 554 139 25.0903
MP2776 lmg/kg
47 990 280 28.2828
48 724 357 49.3094
MEAN 33.4575
STDEV 10.8537
49 421 158 37.5297
MP2776 50
lOmg/kg 51 714 355 49.7199
52 794 245 30.8564
MEAN 39.3687
STDEV 9.5653
53 1028 231 22.4708
MP2814 lmg/kg 54 648 114 17.5926
55 827 278 33.6155 56 1035 171 16.5217
MEAN 22.5502
STDEV 7.8181
57 786 222 28.2443
MP2814 58 747 290 38.822 lOmg/kg 59 688 276 40.1163
60 567 241 42.5044
MEAN 37.4218
STDEV 6.3056
61 656 94 14.3293
62 432 36 8.33333
MP2847 lmg/kg
63 747 102 13.6546
64 715 35 4.8951
MEAN 10.3031
STDEV 4.4933
65 585 51 8.71795
MP2847 66 847 110 12.987 lOmg/kg 67 548 54 9.85401
68 436 48 11.0092
MEAN 10.6420
STDEV 1.8218
69 644 88 13.6646
70 580 102 17.5862
MP2850 lmg/kg
71 692 158 22.8324
72 927 131 14.1316
MEAN 17.0537
STDEV 4.2309
73 1122 344 30.6595
MP2850 74 640 188 29.375 lOmg/kg 75 901 219 24.3063
76 855 326 38.1287
MEAN 30.6174
STDEV 5.7095
77 332 46 13.8554
78 870 52 5.97701
MP2859 lmg/kg
79 654 110 16.8196
80
MEAN 12.2173
STDEV 5.6038
MP2859 81 1242 381 30.6763 21 lOmg/kg 82 415 65 15.6627
21 83 595 97 16.3025
21 84 489 65 13.2924
MEAN 18.9835
STDEV 7.9020
Table 20 - Hepatocyte BrdU positive Nuclei
Treatment Animal (All Analysis Total BrdU Positive % BrdU
/ Dose Group Male) Region Cells Cells Positive Cells
1 1 layer 1 18356 256 1.39464
1 2 layer 2 22595 558 2.46957
Vehicle
1 3 layer 3 14461 185 1.2793
1 4 layer 4 20712 277 1.33739
1.62
0.57
2 5 layer 1 17347 3867 22.292
WT-FGF1 2 6 layer 2 16726 2658 15.8914 lmg/kg 2 7 layer 3 14832 5868 39.5631
2 8 layer 4 13961 5509 39.4599
29.30
12.08
3 9 layer 1 12728 317 2.49057
MP200 3 10 layer 2 11802 583 4.93984 lmg/kg 3 11 layer 3 18185 580 3.18944
3 12 layer 4 15362 413 2.68845
3.33
1.11
4 13 layer 1 14958 704 4.70651
MP200 4 14 layer 2 12395 1391 11.2223
3mg/kg 4 15 layer 3 20147 1228 6.0952
4 16 layer 4 15934 1309 8.21514
7.56
2.84
5 17 layer 1 16338 1478 9.04639
MP200 5 18 layer 2 22644 4395 19.4091 lOmg/kg 5 19 layer 3 13429 2733 20.3515
5 20 layer 4 13844 2202 15.9058
16.18
5.12
Λ/fPO Q'J 6 21 layer 1 12448 2706 21.7384
6 22 layer 2 20221 5356 26.4873 lmg/kg
6 23 layer 3 14531 4656 32.0418 6 24 layer 4 15543 5446 35.0383
28.83
5.91
7 25 layer 1 23298 5615 24.1008
MP293 7 26 layer 2 17050 3341 19.5953 lOmg/kg 7 27 layer 3 24106 8553 35.4808
7 28 layer 4 14080 2415 17.152
24.08
8.13
8 29 layer 1 13136 760 5.78563
MP697 8 30 layer 2 14235 1180 8.28943 lmg/kg 8 31 layer 3 15994 2631 16.4499
8 32 layer 4 15346 4889 31.8585
15.60
11.76
9 33 layer 1 16830 2568 15.2585
MP697 9 34 layer 2 18425 2064 11.2022 lOmg/kg 9 35 layer 3 17288 3077 17.7985
9 36 layer 4 17884 2600 14.5381
14.70
2.72
10 37 layer 1 16216 1043 6.43192
MP 1834 10 38 layer 2 23727 3048 12.8461 lmg/kg 10 39 layer 3 21229 1773 8.35178
10 40 layer 4 16240 1282 7.89409
8.88
2.77
11 41 layer 1 12164 1485 12.2082
MP 1834 11 42 layer 2 18329 2635 14.3761 lOmg/kg 11 43 layer 3 13095 808 6.17029
11 44 layer 4 14097 2350 16.6702
12.36
4.51
12 45 layer 1 15973 3862 24.1783
MP2776 12 46 layer 2 18483 1687 9.12731 lmg/kg 12 47 layer 3 13399 1940 14.4787
12 48 layer 4 13931 4238 30.4214
19.55
9.56
MP2776 13 49 layer 1 16691 572 3.427 lOmg/kg 13 50 layer 2 16341 1165 7.12931
13 51 layer 3 13477 924 6.85613
13 52 layer 4 13329 321 2.40828
4.96 2.39
14 53 layer 1 18748 3275 17.4685
MP2814 14 54 layer 2 16760 1862 11.1098 lmg/kg 14 55 layer 3 16749 2588 15.4517
14 56 layer 4 15091 2907 19.2631
15.82
3.51
15 57 layer 1 18297 2003 10.9471
MP2814 15 58 layer 2 17810 1188 6.67041 lOmg/kg 15 59 layer 3 19638 2048 10.4288
15 60 layer 4 16428 3862 23.5086
12.89
7.33
16 61 layer 1 18045 537 2.97589
MP2847 16 62 layer 2 17423 188 1.07903 lmg/kg 16 63 layer 3 14409 1780 12.3534
16 64 layer 4 13723 535 3.89856
5.08
4.99
17 65 layer 1 23975 1397 5.8269
MP2847 17 66 layer 2 22574 1870 8.28387 lOmg/kg 17 67 layer 3 19500 591 3.03077
17 68 layer 4 17179 897 5.22149
5.59
2.16
18 69 layer 1 17267 761 4.40725
MP2850 18 70 layer 2 22366 7320 32.7282 lmg/kg 18 71 layer 3 17638 3103 17.5927
18 72 layer 4 20195 2713 13.434
17.04
11.82
19 73 layer 1 19825 1729 8.72131
MP2850 19 74 layer 2 20485 1345 6.56578 lOmg/kg 19 75 layer 3 16091 2547 15.8287
19 76 layer 4 18533 2614 14.1046
11.31
4.38 20 77 layer 1 18525 464 2.50472
MP2859 20 78 layer 2 18161 279 1.53626 lmg/kg 20 79 layer 3 12534 1522 12.143
20 80 layer 4 23220 1284 5.52972
5.43
4.79
21 81 layer 1 15705 4456 28.3731
MP2859 21 82 layer 2 15736 1709 10.8604 lOmg/kg 21 83 layer 3 18347 4473 24.38
21 84 layer 4 20121 2226 11.0631
18.67
9.05
[0389] From Tables 17-20 and Figs. 6-9, group 10 (MP1834), group 13 (MP2776), groups 14/15 (MP2814), and groups 16/17 (MP2847) exhibited lower in vivo mitogenicity relative to the other tested FGFl polypeptides.
Example 7
[0390] Assessment of glucose lowering and insulin sensitizing activity in diet induced obese mice (DIO): 21 week old DIO mice were habituated to handling 1 week prior to start of study.
Insulin resistance was confirmed based on 4 hour fasting insulin levels (> 5ng/ml). Study day 1, mice (n=9/group) were dosed subcutaneously with vehicle (Tris/NaCl diluted in PBS), WT-FGF1 or exemplary FGFl variants and examined for changes in blood glucose 4 hours post dose. 24 hours post dose, fed blood glucose, body weight and food intake were monitored. After 14 and 21 days of daily subcutaneous dosing, 4 hour fasting glucose and insulin were measured to determine homeostatic model assessment insulin resistance (HOMA-IR). Mice were not dosed with test article the day before HOMA-IR measurements, to avoid possible effects of acute glucose lowering.
[0391] Figs. 10-12 illustrate the blood glucose level and HOMA-IR measurement in mice tested with exemplary FGFl polypeptides described herein. Fig. 10A, Fig. 11A, and Fig. 12A show the blood glucose level at 0, 4, and 24 hours. Fig. 10B, Fig. 1 IB, and Fig. 12B show HOMA-IR measurement at week 0, week 2, and week 3. FGFl polypeptides showing decreased HOMA-IR relative to week 0 show improved insulin sensitivity. FGFl polypeptides: MP200, MP1348, MP2744, MP2779, and MP2847 show increased sensitivity to insulin at week 2 and week 3 relative to the other tested FGFl polypeptides. Table 21 illustrate the SEQ ID NO. and mutation of respective tested FGFl polypeptides.
Table 21 SEQ ID NO: FGF1 Mutation FGF1 ID
~ Veh Veh
2 WT-FGF1 WT-FGF1
25971 ΔΝΤ9 R88L MP516
25837 ΔΝΤ9 K10E R35E MP2744
25854 ΔΝΤ9 V51A R35E MP2779
25860 ΔΝΤ9 T59A R35E MP2786
25923 ΔΝΤ9 R122A R35E MP2847
25906 ΔΝΤ9 Y125A R35E MP2850
210 ΔΝΤ1-9 Κ118\¥ MP607
50 ΔΝΤΙ-9 K57E MP222
70 ΔΝΤ1-9 E81K MP242
25972 ΔΝΤ1-9 R88D MP518
48 ΔΝΤΙ-9 Y55A MP220
31 ΔΝΤ1-9 R35E MP200
94 ΔΝΤ1-9 Η102Α MP266
25603 FGF1 DNT1-9 R37E/R122A MP1348
[0392] The in-vivo mitogenicity and in-vivo efficacy of exemplary FGF1 polypeptides, MP200 and MP2847, are shown in Fig. 13A - Fig. 13C. MP2847 exhibits reduced liver mitogenicity relative to MP200 and WT FGF1 (Fig. 13A). MP2847 improved insulin sensitivity as shown by decreased HOMA-IR relative to baseline (-56%). MP200 decreased HOMA-IR by -53%, relative to baseline. See Fig. 13B and Fig. 13C.
Example 8
[0393] Glucose Level study: sixty eight week old ob/ob male mice (B6.Cg-Lepob/J, Jackson lab) will be randomized into three groups and treated with daily subcutaneous (s.c.) injection of a FGF mutant described herein, a control drug, or vehicle. Blood glucose level will be measured in fed animals one hour after treatment.
[0394] Glucose tolerance test: glucose tolerance test will be conducted after overnight fasting. Mice will be intraperitoneally (i.p.) injected with about 1 g of glucose/kg body weight, and blood glucose will be monitored at 0, 10, 20, 30, 60, or 120 min post injection. Insulin tolerance tests will be conducted after overnight fasting. Mice will be injected i.p. with 0.5 U of insulin/kg body weight (Humulin R; Eli Lilly) and blood glucose will be monitored at 0, 10, 20, 30, 60, or 120 min post injection.
Example 9 Title: Clinical Trial Study on the Safety and Efficacy of a FGF Mutant for Type 2 Diabetes
Mellitus
[0395] Study Type: Interventional
[0396] Study Design:
[0397] Allocation: Randomized
[0398] Endpoint Classification: Efficacy Study
[0399] Intervention Model: Parallel Assignment
[0400] Masking: Double Blind (Subject, Investigator)
[0401] Primary Purpose: Treatment
[0402] Primary Outcome Measures:
[0403] Change from Baseline in Hemoglobin Ale (HbAlc) at Week 12 [ Time Frame: Baseline,
Week 12 ] [ Designated as safety issue: No ]
[0404] Secondary Outcome Measures:
[0405] Performance of a hyperinsulinemic euglycemic clamp at week 12 [ Time Frame: Baseline, Week 12 ] [ Designated as safety issue: No]
[0406] Change from Baseline in HbAlc at Week 24 [ Time Frame: Baseline, Week 24 ]
[ Designated as safety issue: No ]
[0407] Mean Percent Change from Baseline in Body Weight [ Time Frame: Baseline, Week 12, Week 24 ] [ Designated as safety issue: No ]
[0408] Change from Baseline in Fasting Blood Glucose [ Time Frame: Baseline, Week 12, Week 24 ] [ Designated as safety issue: No ]
[0409] Change from Baseline in 7-point Self-Monitored Blood Glucose (SMBG) Values
[ Time Frame: Baseline, Week 12, Week 24 ] [ Designated as safety issue: No ]
[0410] Change from Baseline in Lipids [ Time Frame: Baseline, Week 24 ]
[ Designated as safety issue: No ]
[0411] Change from Baseline in Fasting Fibroblast Growth Factor 21 [ Time Frame: Baseline, Week 12, Week 24 ] [ Designated as safety issue: No ]
[0412] Percentage of Participants Requiring Rescue Therapy [ Time Frame: Baseline through Therapy Completion (Week 24) ] [ Designated as safety issue: No ]
[0413] Pharmacokinetics: Maximum Concentration (Cmax) of LY2944876 [ Time Frame: Baseline through Therapy Completion (Week 24) ] [ Designated as safety issue: No ]
[0414] Change from Baseline in Adiponectin Levels [ Time Frame: Baseline, Week 12, Week 24 ] [ Designated as safety issue: No ]
[0415] Change from Baseline in Beta-Hydroxy Butyrate Levels [ Time Frame: Baseline, Week 12, Week 24 ] [ Designated as safety issue: No ] [0416] Change from Baseline in Glucagon Levels [ Time Frame: Baseline, Week 12, Week 24 ] [ Designated as safety issue: No ]
[0417] Change from Baseline in Insulin Levels [ Time Frame: Baseline, Week 12, Week 24 ] [ Designated as safety issue: No ]
[0418] Pharmacokinetics: Area Under the Concentration Curve (AUC) of LY2944876
[ Time Frame: Time Frame: Baseline through Therapy Completion (Week 24) ]
[ Designated as safety issue: No ]
[0419] Eligibility:
[0420] Ages Eligible for Study: 18 Years to 80 Years
[0421] Genders Eligible for Study: Both
[0422] Accepts Healthy Volunteers: No
[0423] Criteria
[0424] Inclusion Criteria:
[0425] Men or women with diabetes mellitus Type 2
[0426] Have screening HbAlc >7.0% and <10.5% either on diet and exercise alone or on a stable dose of metformin (>1000 mg/day) for 3 months prior to screening
[0427] Have body mass index (BMI) >23 and <45 kilograms per meter squared at screening
[0428] Exclusion Criteria:
[0429] Women of child bearing potential
[0430] Participants who have used thiazolidinediones within 3 months prior to screening, or any other drugs for treatment of hyperglycemia (except metformin) within the prior 2 months
[0431] Participants who have used insulin for diabetic control for more than 6 consecutive days within the prior year
[0432] Participants with impaired renal function (serum creatinine >124 micromole per liter (μπιοΙ/L) [1.4 milligrams per deciliter (mg/dL)] in women, >133 μπιοΙ/L [1.5 mg/dL] in men)
[0433] The examples and embodiments described herein are for illustrative purposes only and various modifications or changes suggested to persons skilled in the art are to be included within the spirit and purview of this application and scope of the appended claims.

Claims

CLAIMS WHAT IS CLAIMED IS:
1. An isolated and purified FGF polypeptide comprising:
one or more modified amino acids selected from Group 1 amino acid residues: P27, R35, D36, R37, Y55, K57, T61, E81, E87, R88, N92, T96, K101, H102, and L133; and
one or more modified amino acids selected from Group 2 amino acid residues: L26, T30, D39, Q43, Y64, D68, Q77, N80, E82, L84, L86, K105, G120, Q127, and K128;
wherein the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
2. The isolated and purified FGF polypeptide of claim 1, wherein the one or more modified amino acids from Group 1 is selected from Group 1A amino acid residues: R35, D36, and R37.
3. The isolated and purified FGF polypeptide of claim 1, wherein the one or more modified amino acids from Group 1 is selected from Group IB amino acid residues: E87, R88, N92, T96, and L133.
4. The isolated and purified FGF polypeptide of claim 1, wherein the one or more modified amino acids from Group 1 is selected from Group 1C amino acid residues: P27, Y55, K57, T61, E81, K101, and H102.
5. The isolated and purified FGF polypeptide of claim 1, wherein the one or more modified amino acids from Group 1 is R35, N92, or T96.
6. The isolated and purified FGF polypeptide of claim 1, wherein the one or more modified amino acids from Group 1 is R35, and the one or more modified amino acids from Group 2 is selected from Q43, Q77, N80, and E82.
7. The isolated and purified FGF polypeptide of claim 1, wherein the one or more modified amino acids from Group 1 is N92, and the one or more modified amino acids from Group 2 is Q43.
8. The isolated and purified FGF polypeptide of claim 1, wherein the one or more modified amino acids from Group 1 is T96, and the one or more modified amino acids from Group 2 is Q43.
9. The isolated and purified FGF polypeptide of claim 1, wherein the one or more modified amino acids comprise a mutation to a non-polar residue or a charged residue.
10. The isolated and purified FGF polypeptide of claim 1, wherein the one or more modified amino acids comprise a mutation to alanine.
11. The isolated and purified FGF polypeptide of any one of the claims 1-10, further comprising a truncation.
12. The isolated and purified FGF polypeptide of claim 11, wherein the truncation is an N- terminal truncation.
13. The isolated and purified FGF polypeptide of claim 12, wherein the N-terminal truncation is a deletion of amino acid residues 1-9.
14. An isolated and purified FGF polypeptide comprising two or more modifications, the
polypeptide comprising:
a first modification at an amino acid residue selected from: Y15, R35, R37, K57, R88, N92, T96, R122, and L133; and
a second modification at an amino acid residue selected from: Fl, N2, P5, G6, N7, K9, K10, S 17, R35, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, E53, K57, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, T96, 198, K101, E104, K112, S 116, R119, H124, Y125, P136, V137, S 138, S 139, and D140;
wherein the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
15. The isolated and purified FGF polypeptide of claim 14, wherein the second modification is at an amino acid residue selected from: N7, S38, Q43, Q45, A48, S58, T78, P79, N80, E81, E82, E91, H93, 198, K101, P136, and V137.
16. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at R35, and the second modification is at an amino acid residue selected from the group consisting of: Fl, N2, P5, G6, N7, K9, K10, S17, R37, S38, Q40, Q43, Q45, S47, A48, E49, S50, V51, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E91, N92, H93, 198, K101, E104, K112, S116, R119, H124, Y125, P136, V137, S 138, S 139, and D140.
17. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at R35, and the second modification is at an amino acid residue selected from the group consisting of: A48, E91, N92, H93, 198, K101, and V137.
18. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at R35, and the second modification is at an amino acid residue selected from the group consisting of: Fl, N2, P5, K10, R37, Q45, S47, E49, S50, V51, S58, T59, D70, L72, S76, Q77, T78, P79, N80, E81, E82, E90, E104, SI 16, Y125, P136, and S139.
19. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at R35, and the second modification is at an amino acid residue selected from the group consisting of: G6, K9, and S 17.
20. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at N92, and the second modification is at an amino acid residue selected from the group consisting of R37 and Q43.
21. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at R37, and the second modification is at an amino acid residue selected from the group consisting of: Q43, E53, and T96.
22. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at R37, and the second modification is at an amino acid residue selected from the group consisting of N92 and T96.
23. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at R37, and the second modification is at an amino acid residue selected from the group consisting of: Q43 and E53.
24. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at R88, and the second modification is at an amino acid residue selected from the group consisting of E91 and T96.
25. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at K57, and the second modification is at an amino acid residue selected from the group consisting of: K10, A48, E90, and E91.
26. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at K57, and the second modification is at an amino acid residue selected from the group consisting of: K10, A48, and E90.
27. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at T96, and the second modification is at Q43.
28. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at Y15, and the second modification is at Q43.
29. The isolated and purified FGF polypeptide of claim 14, wherein the first modification is at R122, and the second modification is at an amino acid residue selected from the group consisting of: R35, R37, and K57.
30. The isolated and purified FGF polypeptide of any one of claims 14-29, further comprising a truncation.
31. The isolated and purified FGF polypeptide of claim 30, wherein the truncation is an N- terminal truncation.
32. The isolated and purified FGF polypeptide of claim 31, wherein the N-terminal truncation is a deletion of amino acid residues 1-9.
33. The isolated and purified FGF polypeptide of claim 14, wherein the modification comprises a mutation to a non-polar residue or a charged residue.
34. The isolated and purified FGF polypeptide of claim 14, wherein the modification comprises a mutation to alanine.
35. The isolated and purified FGF polypeptide of claim 14, wherein Y15, R35, R37, K57, R88, N92, T96, and R122 are independently mutated to an amino acid selected from the group consisting of: alanine (A), glutamic acid (E), histidine (H), leucine (L), and phenylalanine (F).
36. The isolated and purified FGF polypeptide of claim 14, wherein Y15 is mutated to
phenylalanine (F).
37. The isolated and purified FGF polypeptide of claim 14, wherein R35 is mutated to an amino acid selected from the group consisting of: alanine (A) and glutamic acid (E).
38. The isolated and purified FGF polypeptide of claim 14, wherein R37 is mutated to an amino acid selected from the group consisting of: alanine (A) and glutamic acid (E).
39. The isolated and purified FGF polypeptide of claim 14, wherein K57 is mutated to an amino acid selected from the group consisting of: alanine (A), asparagine (N), glutamic acid (E), and histidine (H).
40. The isolated and purified FGF polypeptide of claim 14, wherein R88 is mutated to leucine (L).
41. The isolated and purified FGF polypeptide of claim 14, wherein N92 is mutated to an amino acid selected from the group consisting of: alanine (A) and threonine (T).
42. The isolated and purified FGF polypeptide of claim 14, wherein T96 is mutated to alanine (A).
43. An isolated and purified FGF polypeptide comprising a modification at amino acid residue A48, E53, K57, S58, L72, S76, Q77, T78, P79, T96, or 198, wherein the numbering of the amino acid residues corresponds to SEQ ID NO: 2.
44. The isolated and purified FGF polypeptide of claim 43, wherein the modification is at K57.
45. The isolated and purified FGF polypeptide of claim 43, wherein the modification is at Q77.
46. The isolated and purified FGF polypeptide of claim 43, wherein the modification is at T96.
47. The isolated and purified FGF polypeptide of any one of the claims 43-46, further comprising a truncation.
48. The isolated and purified FGF polypeptide of claim 47, wherein the truncation is an N- terminal truncation.
49. The isolated and purified FGF polypeptide of claim 48, wherein the N-terminal truncation is a deletion of amino acid residues 1-9.
50. The isolated and purified FGF polypeptide of claim 43, wherein the modification comprises a mutation to a non-polar residue or a charged residue.
51. The isolated and purified FGF polypeptide of claim 43, wherein the modification comprises a mutation to alanine.
52. An isolated and purified FGF polypeptide comprising at least 80% sequence identity to a sequence selected from SEQ ID NOs: 3-25917 and 25922-25974.
53. The isolated and purified FGF polypeptide of claim 52, wherein the FGF polypeptide
comprises at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a sequence selected from SEQ ID NOs: 3-25917 and 25922-25974.
54. The isolated and purified FGF polypeptide of claim 52, wherein the FGF polypeptide consists of a sequence selected from SEQ ID NOs: 3-25917 and 25922-25974.
55. An isolated and purified FGF polypeptide having:
a FGFR interaction activity that is equivalent to the FGFR interaction activity of FGF1 polypeptide of SEQ ID NO: 2; and
a mitogenicity that is at least 1-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2.
56. The isolated and purified FGF polypeptide of claim 55, wherein the FGFR interaction activity of the isolated and purified FGF polypeptide is about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% to the FGFR interaction activity of FGF 1 polypeptide of SEQ ID NO: 2.
57. The isolated and purified FGF polypeptide of claim 55, wherein the mitogenicity is at least 2- fold lower, 3-fold lower, 4-fold lower, 5-fold lower, 6-fold lower, 7-fold lower, 8-fold lower, 9-fold lower, or 10-fold lower than the mitogenicity of the FGF1 polypeptide of SEQ ID NO: 2.
58. The isolated and purified FGF polypeptide of claim 55, wherein the isolated and purified FGF polypeptide is an isolated and purified FGF polypeptide of claims 1-54.
59. The isolated and purified FGF polypeptide of any one of the claims 1-58, wherein the FGF polypeptide is a FGF1 polypeptide.
60. The isolated and purified FGF polypeptide of any one of the claims 1-59, wherein the isolated and purified FGF polypeptide has enhanced thermostability relative to a wild-type FGF polypeptide.
61. The isolated and purified FGF polypeptide of any one of the claims 1-60, wherein the isolated and purified FGF polypeptide has enhanced pH stability relative to a wild-type FGF polypeptide.
62. The isolated and purified FGF polypeptide of any one of the claims 1-61, wherein the isolated and purified FGF polypeptide has reduced or impaired mitogenic activity relative to a wild- type FGF polypeptide.
63. The isolated and purified FGF polypeptide of any one of the claims 1-62, wherein the isolated and purified FGF polypeptide has enhanced FGF receptor interaction relative to a wild-type FGF polypeptide.
64. The isolated and purified FGF polypeptide of any one of the claims 1-63, wherein the isolated and purified FGF polypeptide has increased in vivo half-life relative to a wild-type FGF polypeptide when administered to a subject in need thereof.
65. The isolated and purified FGF polypeptide of any one of the claims 1-64, wherein the isolated and purified FGF polypeptide has increased resistance to proteolysis relative to a wild-type FGF polypeptide.
66. A pharmaceutical composition comprising:
an isolated and purified FGF polypeptide of claims 1-65; and an excipient.
The pharmaceutical composition of claim 66, further comprising at least one additional FGF polypeptide, wherein the at least one additional FGF polypeptide is identical to the FGF polypeptide.
The pharmaceutical composition of claim 66, further comprising at least one additional FGF polypeptide, wherein the at least one additional FGF polypeptide is not identical to the FGF polypeptide.
The pharmaceutical composition of claim 67 or 68, wherein the at least one additional FGF polypeptide comprises a sequence selected from SEQ ID NOs: 3-25917 and 25922-25974. The pharmaceutical composition of any one of the claims 66-69, wherein the at least one additional FGF polypeptide comprises at least one modification, a truncation, or a combination thereof.
The pharmaceutical composition of claim 70, wherein the truncation is an N-terminal truncation, a C-terminal truncation, or a combination thereof.
The pharmaceutical composition of any one of the claims 66-71, wherein the pharmaceutical composition is formulated for oral or parenteral administration.
A method of treating a metabolic disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an isolated and purified FGF polypeptide of claims 1-65, or a pharmaceutical composition of claims 66-72.
The method of claim 73, wherein the metabolic disease or condition comprises FGFl -related disease or condition.
The method of claim 73, wherein the metabolic disease or condition comprises diabetes, polycystic ovary syndrome (PCOS), metabolic syndrome (MetS), obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), dyslipidemia, a cardiovascular disease, Hurler syndrome, Huntington's disease, Alzheimer's disease,
Parkinson's disease, Crohn's disease, Fabry disease, or galactosemia.
The method of claim 73, wherein the metabolic disease or condition comprises impaired insulin sensitivity or glucose intolerance.
The method of claim 73, wherein the subject has diabetes.
The method of claim 77, wherein the diabetes comprises type 1 diabetes, type 2 diabetes, type 3 diabetes, type 4 diabetes, double diabetes, latent autoimmune diabetes (LAD), gestational diabetes, neonatal diabetes mellitus (NDM), maturity onset diabetes of the young (MODY), Wolfram syndrome, Alstrom syndrome, prediabetes, or diabetes insipidus.
The method of claim 77, wherein the subject has type 1 diabetes.
The method of claim 77, wherein the subject has type 2 diabetes.
The method of claim 77, wherein the subject has type 4 diabetes.
The method of claim 77, wherein the subject has pre-diabetes.
83. The method of claim 77, wherein the subject has polycystic ovary syndrome.
84. The method of claim 77, wherein the subject has metabolic syndrome.
85. The method of claim 77, wherein the subject has obesity.
86. The method of claim 77, wherein the subject has non-alcoholic fatty liver disease.
87. The method of claim 77, wherein the subject has insulin resistance.
88. The method of claim 77, wherein the subject has lipodystrophy.
89. The method of claim 77, wherein the subject has a cardiovascular disease.
90. The method of claim 89, wherein the cardiovascular disease comprises hypertension,
coronary artery diseases (CAD), stroke, rheumatic heart disease, hypertensive heart disease, cardiomyopathy, heart arrhythmia, congenital heart disease, carditis, aortic aneurysms, renal artery stenosis, peripheral artery disease, venous thrombosis, heart valve disease, deep vein thrombosis, cerebrovascular disease, cardiac dysrhythmia, chronic obstructive pulmonary disease, or inflammatory heart disease.
91. The method of claim 90, wherein the subject has hypertension.
92. The method of claim 75, wherein the subject has NASH.
93. The method of any one of the claims 73-92, further comprising administration of an
additional therapeutic agent.
94. The method of claim 93, wherein the additional therapeutic agent comprises
thiazolidinedione .
95. The method of claim 94, wherein thiazolidinedione comprises pioglitazone, rosiglitazone, lobeglitazone, and troglitazone.
96. The method of any one of the claims 73-95, wherein the FGF polypeptide and the additional therapeutic agent are administered simultaneously.
97. The method of any one of the claims 73-95, wherein the FGF polypeptide and the additional therapeutic agent are administered sequentially.
98. The method of claim 97, wherein the FGF polypeptide is administered prior to the
administration of the additional therapeutic agent.
99. The method of claim 97, wherein the FGF polypeptide is administered after the
administration of the additional therapeutic agent.
100. The method of any one of the claims 73-96, wherein the FGF polypeptide and the additional therapeutic agent are formulated in a unified dosage form.
101. The method of any one of the claims 73-95 or 97-99, wherein the FGF polypeptide and the additional therapeutic agent are formulated in separate dosage forms.
102. A polynucleic acid molecule encoding a FGF polypeptide of claims 1-65.
103. A kit comprising an isolated and purified FGF polypeptide of claims 1-65.
104. A kit comprising a polynucleic acid molecule encoding a FGF polypeptide of claims 1-65.
PCT/US2017/043383 2016-07-21 2017-07-21 Fgf mutants and uses thereof WO2018018010A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201662365296P 2016-07-21 2016-07-21
US62/365,296 2016-07-21

Publications (1)

Publication Number Publication Date
WO2018018010A1 true WO2018018010A1 (en) 2018-01-25

Family

ID=60992687

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2017/043383 WO2018018010A1 (en) 2016-07-21 2017-07-21 Fgf mutants and uses thereof

Country Status (1)

Country Link
WO (1) WO2018018010A1 (en)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10159711B2 (en) 2010-04-16 2018-12-25 Salk Institute For Biological Studies Methods for treating metabolic disorders using FGF
US20190284252A1 (en) * 2018-03-16 2019-09-19 The Board Of Trustees Of The University Of Arkansas Engineered fgf1 and fgf2 compositions and methods of use thereof
US10695404B2 (en) 2015-10-30 2020-06-30 Salk Institute For Biological Studies Treatment of steroid-induced hyperglycemia with fibroblast growth factor (FGF) 1 analogs
WO2021011878A1 (en) * 2019-07-18 2021-01-21 Venturis Therapeutics, Inc. Method of treating parkinson's disease
CN113194980A (en) * 2018-10-09 2021-07-30 利兰斯坦福初级大学董事会 Engineered fibroblast growth factor variants as receptor antagonists
WO2021239046A1 (en) * 2020-05-28 2021-12-02 江苏康缘瑞翱生物医药科技有限公司 Rhfgf21 fusion protein, polynucleotide encoding rhfgf21 fusion protein, composition containing rhfgf21 fusion protein, and use of rhfgf21 fusion protein
WO2022035849A1 (en) * 2020-08-10 2022-02-17 Venturis Therapeutics, Inc. Methods reducing hair loss and/or increasing hair regrowth
US11542309B2 (en) 2019-07-31 2023-01-03 Salk Institute For Biological Studies Fibroblast growth factor 1 (FGF1) mutant proteins that selectively activate FGFR1B to reduce blood glucose

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150079003A1 (en) * 2012-05-01 2015-03-19 Univeristy of Washington through its Center for Commreicalization Methods and Compositions for Cancer Diagnosis
US20150111821A1 (en) * 2013-10-21 2015-04-23 Salk Institute For Biological Studies Mutated fibroblast growth factor (fgf) 1 and methods of use
US20150361436A1 (en) * 2014-06-17 2015-12-17 Xycrobe Therapeutics, Inc. Genetically modified bacteria and methods for genetic modification of bacteria

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150079003A1 (en) * 2012-05-01 2015-03-19 Univeristy of Washington through its Center for Commreicalization Methods and Compositions for Cancer Diagnosis
US20150111821A1 (en) * 2013-10-21 2015-04-23 Salk Institute For Biological Studies Mutated fibroblast growth factor (fgf) 1 and methods of use
US20150361436A1 (en) * 2014-06-17 2015-12-17 Xycrobe Therapeutics, Inc. Genetically modified bacteria and methods for genetic modification of bacteria

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
LONGO, L: "Experimental Support for the Foldability-Function Tradeoff Hypothesis: Segregation of the Folding Nucleus and Functional Regions in Fibroblast Growth Factor-1", PROTEIN SCIENCE, vol. 21, no. 12, 9 October 2012 (2012-10-09), pages 1911 - 1920, XP055452824 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10159711B2 (en) 2010-04-16 2018-12-25 Salk Institute For Biological Studies Methods for treating metabolic disorders using FGF
US10293027B2 (en) 2010-04-16 2019-05-21 Salk Institute For Biological Studies Methods for treating metabolic disorders using FGF
US10398759B2 (en) 2010-04-16 2019-09-03 Salk Institute For Biological Studies Methods for treating metabolic disorders using FGF
US10695404B2 (en) 2015-10-30 2020-06-30 Salk Institute For Biological Studies Treatment of steroid-induced hyperglycemia with fibroblast growth factor (FGF) 1 analogs
US20190284252A1 (en) * 2018-03-16 2019-09-19 The Board Of Trustees Of The University Of Arkansas Engineered fgf1 and fgf2 compositions and methods of use thereof
US11267855B2 (en) * 2018-03-16 2022-03-08 The Board Of Trustees Of The University Of Arkansas Engineered FGF1 and FGF2 compositions and methods of use thereof
CN113194980A (en) * 2018-10-09 2021-07-30 利兰斯坦福初级大学董事会 Engineered fibroblast growth factor variants as receptor antagonists
WO2021011878A1 (en) * 2019-07-18 2021-01-21 Venturis Therapeutics, Inc. Method of treating parkinson's disease
US11542309B2 (en) 2019-07-31 2023-01-03 Salk Institute For Biological Studies Fibroblast growth factor 1 (FGF1) mutant proteins that selectively activate FGFR1B to reduce blood glucose
WO2021239046A1 (en) * 2020-05-28 2021-12-02 江苏康缘瑞翱生物医药科技有限公司 Rhfgf21 fusion protein, polynucleotide encoding rhfgf21 fusion protein, composition containing rhfgf21 fusion protein, and use of rhfgf21 fusion protein
WO2022035849A1 (en) * 2020-08-10 2022-02-17 Venturis Therapeutics, Inc. Methods reducing hair loss and/or increasing hair regrowth

Similar Documents

Publication Publication Date Title
WO2018018010A1 (en) Fgf mutants and uses thereof
RU2493868C2 (en) Pharmaceutical compositions containing hgh for oral administration
JP2021520391A (en) Pharmaceutical compositions for transmucosal delivery of therapeutic peptides and proteins
TW201116291A (en) Treatments for gastrointestinal disorders
JP6910950B2 (en) Solid oral dosage form
WO2014152090A1 (en) Compositions and treatments of metabolic disorders using fgf binding protein 3 and fgf 19
JP5663741B2 (en) Amino acid conjugated cyanoacrylate polymer particles
US10729636B2 (en) Compositions comprising peptide WKDEAGKPLVK
KR20230110570A (en) Compositions of Peptide Inhibitors of the Interleukin-23 Receptor
JP2019206548A (en) Use of il-22 dimers in manufacture of medicaments for treating pancreatitis
CN108883154A (en) The method for applying hepcidin
CN114302741A (en) Compositions and methods for treating autoimmune disorders
Chen et al. Current therapeutic strategy in the nasal delivery of insulin: recent advances and future directions
CA3103443A1 (en) Methods of making alkaline phosphatase agents
WO2015138638A1 (en) Long acting trail receptor agonists for treatment of autoimmune diseases
US10800812B2 (en) Compstatin analogs with increased solubility and improved pharmacokinetic properties
JP6712230B2 (en) Methods for treating metabolic disorders associated with lipodystrophy and defective insulin production or defective insulin signaling
US20180264085A1 (en) Calcitonin Analogues for Treating Diseases and Disorders
EP1392347B1 (en) Preventing cell death using segments of neural thread proteins
US10010580B2 (en) Insulin independence among patients with diabetes utilizing an optimized hamster Reg3 gamma peptide
JP2023116456A (en) Composition and method for treating or ameliorating scleroderma-associated vasculopathy
JP2017533967A (en) Method for treating Huntington&#39;s disease using cysteamine composition
EP3329930A1 (en) Pharmaceuctical compositions
Twarog Comparison of the intestinal permeation enhancers, SNAC and C₁₀, for oral peptides: biophysical, in vitro and ex vivo studies
JP2019520341A (en) Compositions and methods for treating metabolic disorders

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17831994

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17831994

Country of ref document: EP

Kind code of ref document: A1