WO2011068667A1 - Pyrazolopyrimidines and related heterocycles as ck2 inhibitors - Google Patents

Abstract

The invention provides compounds that inhibit protein kinase CK2 activity (CK2 activity), and compositions containing such compounds. These compounds and compositions are useful for treating proliferative disorders such as cancer, as well as other kinase associated conditions including inflammation, pain, and certain immunological disorders, and have the following general formula (I)..

Description

PYRAZOLOPYR1 IDINES AND RELATED HETEROCYCLES AS

CK2 INHIBITORS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the bene ill of U.S. Provisional Application No. 61/266,801 , filed December 4, 2009 and entitled "PYRAZOLQPYR1MIDINES AND RELATED

HETEROCYCLES AS KINASE INHIBITORS"; and U.S. Provisional Application No.

61/354,165, filed June 11, 2010 and entitled "PYRAZOLGPYRIMIDINES AND RELATED HETEROCYCLES AS CK2 INHIBITORS"; the contents of which are hereby incorporated by references in their entireties for all puiposes.

FIELD OF THE INVENTION

[0002] The invention relates in part to molecules having certain biological activities that include, but are not limited to, inhibiting cell proliferation, and modulating certain protein kinase activities. Molecules of the invention modulate, e.g., Protein Kinase CK2 (called CK2 herein) and are useful to treat conditions associated directly or indirectly with CK2 activities, e.g., cancers, inflammatory conditions, infectious disorders, pain, immunological disorders, a neurodegenerative disorder (such as Alzheimer's disease and Parkinson^'s disease), etc, The invention also relates in part to methods for using such compounds, and pharmaceutical compositions containing these compounds.

BACKGROUND

[0003] Protein kinase CK2 (formerly called Casein kinase II, referred to herein as "CK2") is a ubiquitous and highly conserved protein serine/threonine kinase. The holoenzyme is typically found in tetrameric complexes consisting of two catalytic (alpha arid/or alpha') subunits and two regulatory (beta) subunits. C 2 has a number of physiological targets and participates in a complex series of cellular functions including the maintenance of cell viability. The level of CK2 in normal cells is tightly regulated, and it has long been considered to play a role in cell growth and proliferation. Inhibitors of CK2 that are useful for treating certain types of cancers are described in PCT/US2007/077464, PCT/US2008/074820, PCT/US2009/35609.

[0004] The prevalence and importance of CK2, as well as an evolutionary analysis of its sequence, suggest it is an ancient enzyme on the evolutionary scale; its longevity may explain why it has become important in so many biochemical processes, and why CK2 from hosts have even been co-opted by infectious pathogens (e.g., viruses, protozoa) as an integral part of their survival and life cycle biochemical systems. These same characteristics explain why inhibitors of CK2 are believed to be useful in a variety of medical treatments as disc ussed herein. Because CK2 is centra! to many biological processes, as summarized by Guerra & Issinger, Curr. Med. Chem. , 2008, 15: 1870-1886, inhibitors of CK2, including the compounds described herein, should be useful in the treatment of a variety of diseases and disorders.

[0005] Cancerous cells show an elevation of CK2, and recent evidence suggests that CK2 exerts potent suppression of apoptosis in cel!s by protecting regulatory proteins from caspase- mediated degradation. The anti-apoptotic function of C 2 may contribute to its ability to participate in transformation and tumorigenesis. In particular, CK2 has been shown to be associated with acute and chronic myelogenous leukemia, lymphoma and multiple myeloma. In addition, enhanced CK2 activity has been observed in solid tumors of the colon, rectum and breast, squamous cell carcinomas of the lung and of the head and neck (SCCHN),

adenocarcinomas of the lung, colon, rectum, kidney, breast, and prostate. Inhibition of CK2 by a small molecule is reported to induce apoptosis of pancreatic cancer cells, and hepatocellular carcinoma cells (HegG2, Hep3, HeLa cancer ceil lines); and CK2 inhibitors dramatically sensitized RMS (Rhabdomyosarcoma) tumors toward apoptosis induced by TRAIL. Thus an inhibitor of CK2 alone, or in combination with TRAIL or a ligand for the TRAIL receptor, would be useful to treat RMS, the most common soft-tissue sarcoma in children. In addition, elevated CK2 has been found to be highly correlated with aggressiveness of neoplasias, and treatment with a CK2 inhibitor of the invention should thus reduce tendency of benign lesions to advance into malignant ones, or for malignant ones to metastasize.

[0006] Unlike other kinases and signaling pathways, where mutations are often associated with structural changes that cause loss of regulatory control, increased CK2 acti vity le vel appears to be generally caused by upregulation or overexpression of the active protein rather than by changes that affect activation levels. Guerra and Issinger postulate this may be due to regulation by aggregation, since activity levels do not correlate well with mRNA levels.

Excessive activity of CK2 has been shown in many cancers, including SCCHN tumors, lung tumors, breast tumors, and others. Id.

[0Θ07] Elevated CK2 activity in colorectal carcinomas was shown to correlate with increased malignancy. Aberrant expression and activity of C 2 have been reported to promote increase nuclear levels of NF-kappaB in breast cancer cells, CK2 activity is markedly increased in patients with AML and CML during blast crisis, indicating that an inhibitor of CK2 should be particularly effective in these conditions, Multiple myeloma cell survival has been shown to rely on high activity of C 2, and inhibitors of CK2 were cytotoxic to MM cells.

[0008] The literature provides clear evidence that inhibition of CK2 correlates with efficacy against tumor cells. For example, a C 2 inhibitor inhibited growth of murine p i 90 lymphoma cells. Its interaction with Bcr/Abl has been reported to play an important role in proliferation of Bcr/Abi expressing cells, indicating inhibitors of CK2 may be useful in treatment of Bcr/Abl- positive leukemias. Inhibitors of CK2 ha ve been shown to inhibit progression of skin papillomas, prostate and breast cancer xenografts in mice, and to prolong survi val of transgenic mice that express prostate-promoters. Id.

[0009] The role of CK2 in various non-cancer disease processes has been recently reviewed. See Guerra & Issinger, Curr. Med. Chem., 2008, 15:1870-1886. Increasing evidence indicates that CK2 is in vol ved in critical diseases of the central nervous system, including, for exam ple, Alzheimer's disease, Parkinson's disease, and rare neurodegenerative disorders such as Guam- Parkinson dementia, chromosome 18 deletion syndrome, progressive supranuclear palsy, Kuf s disease, or Pick's disease. It is suggested that selective CK2 -mediated phosphorylation of tau proteins may be involved in progressive neurodegeneration of Alzheimer's disease. In addition, recent studies suggest that CK2 plays a role in memory impairment and brain ischemia, the latter effect apparently being mediated by CK2's regulator}' effect on the PBK survival pathways.

[0010J CK2 has also been shown to be involved in the modulation of inflammatory disorders, for example, acute or chronic inflammatory pain, glomerulonephritis, and

autoimmune diseases, including, e.g., multiple sclerosis (MS), systemic lupus erythematosus, rheumatoid arthritis, and juvenile arthritis. It positively regulates the function of the serotonin 5- HT3 receptor channel, activates heme oxygenase type 2, and enhances the activity of neuronal nitric oxide synthase. A selective CK2 inhibitor w^ras reported to strongly reduce pain response of mice when administered to spinal cord tissue prior to pain testing. It phosphorylates secretory type I I A phospholipase A2 from synovial fluid of RA patients, and modulates secretion of DE (a nuclear DNA-binding protein), which is a proinflammatory molecule found in synovial fluid of patients with juvenile arthritis. Thus, inhibition of CK2 is expected to con trol progression of inflammatory pathologies such as those described here, and the inhibitors disclosed herein have been shown to effectively treat pain in animal models.

[0011] Protein kinase CK2 has also been shown to play a role in disorders of the vascular system, such as, e.g., atherosclerosis, laminar shear stress, and hypoxia. CK2 has also been shown to play a role in disorders of skeletal muscle and bone tissue, such as cardiomyocyte hypertrophy, impaired insulin signaling and bone tissue mineralization, In one study, mhibitors of CK2 were effective at slowing angiogenesis induced by growth factor in cultured cells. Moreover, in a retinopathy model, a CK2 inhibitor combined with octreotide (a somatostatin analog) reduced neovascular tufts; thus, the C 2 inhibitors described herein would be effective in combination with a somatostatin analog to treat retinopathy.

[0012] CK2 has also been shown to phosphor late GSK, troponin and myosin light chain; thus, CK2 is important in skeletal muscle and bone tissue physiology, and is linked to diseases affecting muscle tissue.

[0013] Evidence suggests that CK2 is also involved in the development and life cycle regulation of protozoal parasites, such as, for example, Theileria parva, Trypanosoma cruzi, Leishmania donovani, Herpetomonas muscarum muscarum, Plasmodium falciparum,

Trypanosoma brucei, Toxoplasma gondii and Schistosoma mansoni. Numerous studies have confirmed the role of CK2 in regulation of cel lular motility of protozoan parasites, essential to invasion of host ceils. Activation of CK2 or excessive activity of CK2 has been shown to occur in hosts infected with Leishmania donovani, Herpetomonas muscarum muscarum, Plasmodium falciparum, Trypanosoma brucei, Toxoplasma gondii and Schistosoma mansoni. Indeed, inhibition of CK2 has been sho wn to block infection by T. cruzi.

[ 0014] CK2 has also been shown to interact with and/or phosphorylate viral proteins associated with human immunodeficiency virus type 1 (HIV-1 ), human papilloma virus, and herpes simplex virus, in addition to other virus types (e.g. human cytomegalovirus, hepatitis C and B viruses, Boraa disease virus, adenovirus, coxsackievirus, coronavirus, influenza, and varicella zoster vims). CK2 phosphorylates and activates HIV-1 reverse transcriptase and proteases in vitro and in vivo, and promotes pathogenicity of simian-human immunodeficiency virus (SHIV), a model for HIV. Inhibitors of CK2 are thus able to reduce pathogenic effects of a model of HIV infection. CK2 also phosphorylates numerous proteins in herpes simplex virus and numerous other viruses, and some evidence suggests viruses have adopted CK2 as a phosphorylating enzyme for their essential life cycle proteins. Inhibition of CK2 is thus expected to deter infection and progression of viral infections, which rely upon the host's CK2 for their own life cycles.

[0015] CK2 is unusual in the diversity of biological processes that it affects, and it differs from most kinases in other ways as well: it is constitutive ly active, it can use ATP or GTP, and it is elevated in most tumors and rapidly proliferating tissues. In addition, while many kinase inhibitors affect multiple kinases, increasing the likelihood of off-target effects or variability between individual subjects, CK2's unique structural features enable discovery of highly CK2- specific inhibitors. For all of these reasons, CK2 is a particularly interesting target for drug development, and the invention provides highly effective inhibitors of CK2 that are useful in treating a variety of different diseases and disorders mediated by or associated with excessive, aberrant or undesired levels of CK2 activity.

[001 ] Compounds of Formula I have been found to be active on C 2 as well as on one or more Pim proteins. It has now been found that compounds of Formula (II) and (IF) are typically more active on CK2, and also have less activity on Pim kinases. Without being bound by the theory, it is believed that their physiological activities derive from their activity on CK2.

[0017] The current invention provides novel compounds of Formula (II) and (IF), as well as Formulae Ila, Ila', II-Th and II-Th', and pharmaceutical compositions containing these compounds. The novel compounds of Formula II, which are related to the compounds of Formula L show surprisingly greater activity on CK2 and reduced Pim activity, and thus are advantageously used to treat conditions sensitive to CK2 inhibition such as those described herein. Compounds of Formula II are therefore useful to treat conditions mediated by or associated with excessive activity of CK2, with reduced likelihood of off-target effects caused by inhibition of other kinases.

DISCLOSURE OF THE INVENTION

[0018] The present invention in part provides chemical compounds having certain biological activities that include, but are not limited to, inhibiting cell proliferation, inhibiting

angiogenesis, and modulating protein kinase activities. These molecules modulate protein kinase CK2 (CK2) and/or PIM activity, and are typically more selective for CK2 activity over other kinases than similar compounds that lack the amine group shown in Formula (II) or (IF). These compounds affect biological functions that include but are not limited to, inhibiting gamma phosphate transfer from ATP to a protein or peptide substrate, inhibiting angiogenesis, inhibiting cell proliferation and inducing cell apoptosis, for example. The present invention also in part provides methods for preparing novel chemical compounds, and analogs thereof and methods of using these compounds. Also provided are compositions comprising these molecules in combination with other materials, including other therapeutic agents, and methods for using such compositions. [0019] Compounds of the general formula (I) have been shown to inhibit Pirn and CK2 (PCT/US2010/035657):

wherein the bicyclic ring system containing Z*-Z⁴ is aromatic;

one of Z^¾ and Z² is C, the other of Z' and Z² is N;

Z³ and Z⁴ are independently CR⁵ or N,

where R³ can be H or R^J ;

R¹ is H, halo, CN, optionally substituted C1-C4 alkyl, optionally substituted C2~ C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C 1-C4 alkoxy, or -NR⁷R^s,

where R' and R^' are each independently selected from H, optionally substituted CI -CIO alkyl, optionally substituted aryl, optionally substituted aiylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl,

or R' and R⁸ taken together with the N of -NR^?R⁸ form an optionally substituted 5-8 membered ring that optionally contains a additional heteroatom selected from N, O and S as a ring member;

R² is H, halo, CN, or an optionally substituted group selected from C1-C4 alkyl, C2-C4 alkenyl, and C2-C4 alkynyl;

R³ and R⁴ are independently selected from H and optionally substituted CI -CIO alkyl;

X is NR⁶, O, or S, where R⁶ is H or an optionally substituted group selected from C1-C4 alkyl, C2-C4 alkenyl, and C2-C4 alkynyl;

Y is O or S;

W is optionally substituted aryl, optionally substituted heteroaryl, or ~NR Rⁱ⁰, - OR⁹, S(0)_nR⁹, optionally substituted carbon-linked heterocyclyl, optionally substituted

C3-C8 cycioalkyl, or CR⁹R¹⁰Rⁿ, wherein n is 0, 1 or 2, and

R⁹ and R^R' are each independently selected from H, optionally substituted Cl- C10 alkyl, optionally substituted aryl, optionally substituted arylaSkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl,

or R⁹ and R¹⁰ taken together with the N of -NR⁹R¹⁰ form an optionally substituted 5-8 membered ring that optionally contains an additional heteroatom selected from N, O and S as a ring member, and

R¹¹ is selected from H, optionally substituted C1-C10 alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substi tuted he teroary 1 alkyl ,

[0020] The compounds of Formula I inhibit Pirn and CK2, and often inhibit other kinases as well. For use as pharmaceuticals, it can be advantageous to select compounds that inhibit one primary target enzyme or receptor with minimal affect on other pathways or targets, because off- target biochemical effects can cause unpredictable side effects, It has now been found that compounds of Formula (II) and (IF), which are related to the compounds of Formula I, retain high levels of CK2 activity, and indeed are often more potent on CK2 than other compounds like Formula I, yet they are typically selective for C 2 over Pirn kinases. In addition, their selectivity for CK2 over other kinases in a broad array of kinases is also improved over that of the compounds of Formula I generally. Therefore, compounds of Formula (II) or (IF) represent a particularly useful class of compounds for the methods of treatment described herein, because they are selective for CK2 and inhibit fewer other kinases, resulting in a reduced risk of side- effects.

(II) (IF)

wherein: Z^J and Z⁴ each independently represent N or CR:\ or CH;

each R³ is independently selected from halo, CN, R, -OR, -S(0)_nR, COOR, CONR², and NR₂,

wherein each R is independently selected from H and optionally substituted C l - C4 alkyl, and the two R groups of NR₂ can be linked together to form a 5-6 membered heterocyclic ring that is optionally substituted and can include an additional heteroatoni selected from N, O and S as a ring member;

R ^_, R^J and R⁴ are each independently selected from H and optionally substituted alkyl;

X represents O, S, or NR ;

Y is O or S or NR^{! 0};

where R^1J is selected from H, CN, optionally substituted C 1 -C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, and -NR⁷R⁸,

Z i s O or 8;

L can be a bond,

~(CR⁷R⁸)_m~NR⁷~, -( C;R ^"R^': ) ,-( )-. or -{CR⁷R⁸)_m~S(0)_n~;

W is optionally substituted CI -CIO alkyl, optionally substituted aryi, optionally substituted heteroaryl, -NR⁷R⁸, -OR⁷, S(0)_nR⁷, CONR⁷R⁸, optionally substituted heterocyclyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C10 alkynyl, or CR 'R⁸R⁹,

where each R⁷ and R⁸ and R⁹ is independently selected from H, optionally substituted C1-C6 alkoxy, optionally substituted C 1-C6 alkylamino, optionally substituted C1-C6 dialkylamino, optionally substituted heterocyclyl, optionally substituted CI -CIO alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C4-C10 cycloalkylaikyi, optionally substituted aryi, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylaikyi;

or R⁸ and R⁹ taken together can be =0 (oxo) or ^:::N-OR ' or =N-CN;

or R' and R⁸ taken together with the N of -NR⁷R⁸ can form an optionally substituted 5-10 membered heterocyclic or heteroaromatic ring system that optionally contains an additional heteroatoni selected from N, O and S as a ring member;

provided that no more than one of or R' and R in -NR^'R is selected from the group consisting of alkoxy, alkylamino, dialkylamino and heterocyclyl; each n is independently is 0, 1 or 2;

each in is independently 1 , 2, 3 or 4;

R^1A and R^llJ are each independently selected from H, optionally substituted Cl- C10 alley!, optionally substituted heterocvclyl, optionally substituted cycloalkyL optionally substituted cycloalkylalkyl, optionally substituted heterocyclylalkyl, optionally substituted arylalkyl, or an optionally substituted 5-6 membered aryl ring containing up to two heteroatonis as ring members;

or R^lA and R^iB in -NR^!AR^iB can be taken together to form an optionally substituted 5-8 membered monocyclic or 5- 10 membered bicyclic heteroaryl or heterocyclic group containing up to two additional heteroatoms selected from N, O and as ring members;

and pharmaceutically acceptable salts of these compounds.

21] A favored class of compounds of Formula II are those of Formula (Ha) or (Ila'):

(Ila) i l ia^" )

where R² is H, Me or CF ; R⁴ is H, Me or CF₃; X is O, S or N i l: Y is O or S; R^1B is as described for Formula II; L is a bond, -NR.⁷-, -0-, or -S(0)_ri-, (CR⁷R⁸)_m, or it can be -(CR⁷R^s)_m- NR⁷-; m is 1-4 and n is 0-2: and W is selected from optionally substituted aryl, optionally

7 8 " 8

substituted heteroaryl, and -NR R^' , where R^' and R are as defined for Formula I L [0022] Particular embodiments of the compounds of the invention include thiophene- containing compounds of Formula (II-Th) and (II-Th'):

or

(II-Th) (II-Th')

where R^{J h} is selected from H, halo, optionally substituted C1-C6 alkyi, CN, S(0)o-₂R, -SU NK,.. COOR, CONK,., and C(0)R,

where each R is independently H, halo, CN, or an optionally substituted member selected from the group consisting of C1-C6 alkyi, CI-C6 alkoxy, C1-C6 alkylamino, di(Cl-C6)alkyiamino, C3-C8 cycloalkyl, C4-C10 cycloalkyl alkyi, C5-C8 heterocyclyi, C6-C10 heterocyclylalkyl, aryl, aryialkyl, C5-C6 heteroaikyi, and C6-C10

heteroalkyl alkyi;

and two R on the same atom or adjacent atoms can form an optionally substituted heterocyclic ring that can contain an additional heteroatom selected from N, O and S; and other structural features are as defined for Formula lia above.

[0023] The invention includes pharmaceutically acceptable salts of compounds of

Formula II, IF, Ila, Ha', Ιί-Th, and II-Th' as well as the neutral compounds.

[0024] The invention also provides pharmaceutical compositions containing such compounds plus one or more pharmaceutically acceptable carriers or excipients, and methods of using these compounds and compositions for the treatment of specified conditions as further described herein.

[0025] In addition, the invention provides intermediates of Formula (III), which are useful for preparation of compounds described above, and methods of using these intermediates to make compounds of Formula (II):

where R^{l f}\ ^{^i h}, R², R.\ Z³, Z L and W are as defined for Formula (il) above, or in certain embodiments, these are the same as the corresponding features defined for Formula (Ila) above;

one of Z¹ and 7 represents N, and the other of Z¹ and 7 represents C;

and the circles inside the two rings indicate that the rings are both aromatic.

[0026] The method comprises reacting a compound of Formula (III) with a hydantoin or similar 5-membered heterocyclic compound of Formula (IV):

where R^", X, Y and Z are as defined for Formula (II) or (If ),

under conditions that promote condensation of the two compounds,

[0027] Typically, the reaction conditions will include a suitable solvent and a base, optionally a catalytic amount of base, but stoichiometric or larger amounts of base can be used. Suitable bases are those capable of deprotonating the compound of Formula (IV) to promote condensation with the compound of Formula (IK), and secondary amines that are capable of reacting with aldehydes of Formula (III) to form an iminium species. Suitable bases include C1 -C4 alkoxides, metal hydrides, tertiary amines such as triethylamine or diisopropyl ethylamine, DABCO, DBU and the like; and suitable secondary amine bases include piperidine, morpholine, piperazine, N-methySpiperazine, pyrrolidine, and the like. Suitable solvents include polar aprotic solvents such as NMP, DMF, DMSO, DMA, and dioxane; as well as protic solvents such as C l -C l O alcohols and diols, e.g., ethanol, propanol, isopropanol, ethylene glycol, propylene glycol , methoxyethanol, and the like. Mixtures of such solvents ca also be used, as can mixtures of one or more of these solvents with a less polar organic solvent to promote solubility of the reactants. Selection of suitable solvents and bases for these reactions are well within the level of skill of an ordinary practitioner.

[0028] In some embodiments of the compounds of Formula (III), -L-W represents a group of the formula ~-S(0)_1-2R, where R is an alkyl, cycloalkyl, aryl, heteroaryl or similar group, and the product is a compound of Formula (II) or (IF) having the same -L-W group. Such compounds are conveniently used for the preparation of other compounds of formula (II) or (IV), because the moiety of formula -S(0)_1-2R is a good leaving group, and can readily be displaced by nucleophiles such as primary or secondary amines, to introduce other -L-W groups. Thus another method for synthesizing the compounds of the invention is to react a compound of Formula (V),

wherein -L-W represents a group of the formula ~S(0)i-₂R, where R is an optionally substituted group selected from C1-C6 alkyl, C3-C8 cycloalkyl, C4-C10 cycloalkyl alkyl, C6-C10 aryl, C5-C6 heteroaryl, C7-C12 arylalkyl, and C6-C12 heteroarylaikyl;

and other variables are as defined for formulas (ill) and (IV) above; with a nucleophilic compound of formula

W'-L'-H

wherein L' is selected from NR ', O and S; and

W' is optionally substituted aryl, optionally substituted heteroaryl optionally substituted heterocyclyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C10 alkerryl, optionally substituted C2-C10 alkynyl, or CR' R⁸R⁹,

where R' , R^s and R⁹ are as defined above for Formula I I under suitable conditions as described herein to ro vide a compound of Formul

[0029] Also provided herein are pharmaceutical compositions comprising a compound of Formula I or II as described herein and at least one pharmaceutically acceptable carrier or excipient, or two or more pharmaceutically acceptable carriers and/or excipients.

Pharmaceutical compositions comprising at least one of these compounds can be utilized in methods of treatment such as those described herein.

[0030] The compounds of Formulae i and II as described herein bind to and inhibit certain kinase proteins, which is believed to be the basis for their pharmaceutical activity. In certain embodiments, the protein is a C 2 protein, such as a CK2 protein comprising the amino acid sequence of SEQ ID NO: l, 2 or 3 or a substantially identical variant thereof for example.

SEP ID NO: l ΓΝΡ 001 886; casein kinase H alpha 1 subunit isoform a [Homo sapiens!)

msgpvpsrar vytdvnthrp reywdyeshv vewqnqddyq lvrklgrgky sevfeain.it nnekvvvkii kpvkkkkikr eikilenlrg gpniitladi vkdpvsrtpa lvfehvnntd 121 fkqlyqtitd ydirfymyei lkaldychsra gimhrdvkp nvmidhehrk Irlidwglae 181 fyh gqeynv rvasryfk.gp ellvdyqmyd ysldmwslgc mlasmifrke pffhghdnyd 241 qlvriakvlg tedlydyidk ynieldprfn dilgrhsrkr werfvhsenq hlvspealdf 301 Idkllrydhq srltareame hpyfytvvkd. qarmgsssmp ggstpvssan mmsgissvpt 361 psplgplags pviaaanplg rapvpaaagaq q SEP ID NO:2 (NP 808227; casein kinase II alpha 1 subunit isoform a [Homo sapiens!)

msgpvpsrar vytdvnthrp reywdyeshv vewgnqddyq lvrklgrgky sevfeainit nnekvvvkii kpvkkkkikr eikilenlrg gpniitladi vkdpvsrtpa lvfehvnntd

121 fkqlyqtitd ydirfymyei lkaldychsra gimhrdvkph nvmidhehrk Irlidwglae

181 fyhpgqeynv rvasryfkgp ellvdygmyd ysldmwslgc mlasmifrke pffhghdnyd 241 qlvriakvlg tedlydyidk ynieldprfn dilgrhsrkr werfvhsenq hlvspealdf

301 Idkllrydhq srltareame hpyfytvvkd qarmgsssmp ggstpvssan mmsgissvpt

361 psplgplags pviaaanplg mpvpaaagaq q SEQ ID NO; 3_. (NP 808228 ; casein kinase II alpha 1 subunit isoforrn b [Homo sapiens]) rayei1ka1ly chsmgimhrd vkphnvmidh e rklr1idw glaefy pgq eynvrvasry fkgpel lvdy qmydys1ditw slgcmlasmi frkepffhg dnydql ria kvlgtedlyd

121 yidkynield prfndilgrh srkrwerfvh senqhl spe ald.fld.kllr ydhqsrltar 181 eame pyfyt vkdqaritgs ssmpggstpv ssanmmsgis svptpsplgp lagspviaaa

241 nplgmpvpaa agaqq

[0031] Substantially identical variants of these include proteins having at least 90% sequence homology with one of these, preferably at least 90% sequence identity; and having at least 50% of the level of in vitro kinase activity of the specified sequence under typical assay conditions,

[0032] The invention includes methods to modulate the activity of CK2 protein, either in vitro or ex vivo, Suitable methods comprise contacting a system comprising the protein with a compound described herein in an amount effective for modulating the activity of the protein, In certain embodiments the activity of the protein is inhibited, and sometimes the protein is a CK2 protein comprising the amino acid sequence of SEQ ID NO: 1 , SEQ ID NO:2 or SEQ ID NO:3 or a substantially identical variant thereof, for example. In certain embodiments the CK2 is in a cell or tissue; in other embodiments, it can be in a cell-free system.

[0033] Provided also are methods for inhibiting cell proliferation, which comprise contacting cells with a compound described herein in an amount effective to inhibit proliferation of the cells. The cells sometimes are in a cell line, such as a cancer cell line (e.g., breast cancer, prostate cancer, pancreatic cancer, lung cancer, hemopoietic cancer, colorectal cancer, skin cancer, ovary cancer cell line), for example, In some embodiments, the cancer cell line is a breast cancer, prostate cancer or pancreatic cancer cell line. The cells sometimes are in a tissue, can be in a subject, at times are in a tumor, and sometimes are in a tumor in a subject. In certain embodiments, the method, further comprises inducing cell apoptosis. Ceils sometimes are from a subject having macular degeneration.

[0034] Also provided are methods for treating a condition related to aberrant cell proliferation, which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the cell proliferative condition, In certain embodiments the cell proliferative condition is a tumor-associated cancer, e.g., a solid or circulating tumor. The cancer sometimes is cancer of the breast, prostate, pancreas, lung, colorectum, skin, or ovary, In some embodiments, the cell proliferative condition is a non-tumor cancer, such as a hematopoietic cancer, for example, including leukemias, e.g., multiple myeloma and

lymphomas. The cell proliferative condition is macular degeneration in some embodiments. [0035] The invention also includes methods for treating cancer or an inflammatory disorder or other disorders described herein that are mediated by excessive activity of one or more of these kinases, in a subject in need of such treatment, comprising: administering to the subject a therapeutically effective amount of a therapeutic agent useful for treating such disorder; and administering to the subject a molecule described herein , e.g., a compound inhibits CK2 in an amount that is effective to enhance a desired effect of the therapeutic agent. In certain embodiments, the molecule that inhibits CK2 is a compound of Formula I or Formula II, or Formula IF or ( Ha) or (Ila'), or a pharmaceutically acceptable salt thereof. In certain embodiments, the desired effect of the therapeutic agent that is enhanced by the molecule that inhibits CK2 is an increase in apopiosis in at least one type of cell, In certain embodiments, the cell is a cancer cell and the compound is a compound of Formula (II) or (Ila) that is a potent inhibitor (IC-50 less than about 100 M, for example) of CK2, Preferably, the compound has an IC-50 on Pirn of less than about 30 nM, and is selective for CK2 over Pirn kinases. In certain embodiments, the IC-50 for inhibition of CK2 is lower by at least a factor of ten than activity on Pirn; in preferred embodiments, the compound has an IC-50 for CK2 that is lower than its IC-50 for at least one of Pirn- 1, Pini-2 and Pim-3 by about 100-fold or more.

[0036] in some embodiments, the therapeutic agent and the molecule that inhibits C 2 are administered at substantially the same time. The therapeutic agent and molecule that inhibits CK2 sometimes are used concurrently by the subject, The therapeutic agent and the molecule that inhibits CK2 can be combined into one pharmaceutical composition in certain

embodiments; in other embodiments that are admistered as separate compositions,

[0037] Also provided are compositions of matter comprising a compound described herein and an isolated protein. The protein sometimes is a CK2 protein, such as a CK2 protein comprising the amino acid sequence of SEQ ID N(): l, SEQ ID NO:2 or SEQ ID NO:3 or a substantially identical variant thereof, for example. In some embodiments, the protein is a Pirn protein. Certain compositions comprise a compound described herein in combination with a cell. The cell may be from a cell line, such as a cancer cell line. In the latter embodiments, the cancer cell line is sometimes a breast cancer, prostate cancer, pancreatic cancer, lung cancer, hematopoietic cancer, colorectal cancer, skin cancer, of ovary cancer cell line.

[0038] These and other embodiments of the invention are described in the description that follows. BRIEF DESCRIPTION OF THE FIGURES

[0039] Figure 1 depicts a compound of Formula I as described herein, and shows its IC50 on CK2 (7 nM) and on PIMl (351 nM), and also shows a plot of inhibition of a panel of 108 kinases to illustrate its selectivity for these kinases relative to other kinases.

[0040] Figure 2 depicts a compound of Formula II as described herein, and shows that it is more potent on CK2 (3 nM), less potent on PIMl (1310 nM), and generally more selective towards various kinases than is the compound in Figure 1 ,

[0041] Figure 3 shows a syn thesis scheme for preparing certain compounds of the invention containing a thiophene ring.

[0042] Figure 4 illustra tes the syntheses of certain pyrazolotriazines of the invention.

[0043] Figure 5 illustrates synthesis methods for introducing various nucleophilic groups onto a pyrazolo-triazine ring system for preparing compounds of the invention.

[ 0044] Figure 6 illustrates general synthesis routes for making certain pyrazolo-triazine compounds of the invention.

[0045] Figure 7 shows a general synthetic method for making various imidazo-pyrazine ring systems andfor making certain compounds of the invention.

[0046] Figure 8 depicts a number of variations of the pyrazolo-triazine compounds within the scope of the invention.

[0047] Figure 9 depicts methods to make certain imidazo-pyridazine compounds within the scope of the invention.

[0048] Figure 10 illustrates a general method for modifying certain substituted compounds of the in venti on to introduce additional features.

[0049] Figure 11 depicts more methods for modifying substituents on compounds of the invention.

[0050] Figure 12 illustrates alternative synthesis routes for making certain compounds of the invention.

[0051] Figure 13 depicts formation of an amide compound of the invention from a corresponding carboxylic acid compound.

[0052] Figure 14 depicts a reductive animation method for introducing certain groups onto the compounds of the invention. MODES OF CARRY ING OUT THE INVENTION

[0053] Compounds of the present invention exert biological activities that include, but are not limited to, inhibiting cell proliferation, reducing angiogenesis, preventing or reducing inflammatory responses and pain, and modulating certain immune responses. Such compounds modulate CK2 activity, as demonstrated by the data herein. Such compounds therefore can be utilized in multiple applications by a person of ordinary skill in the art. For example, compounds described herein can be used, for example, for (i) modul ation of protein kinase activity (e.g., CK2 activity), (ii) modulation of cell proliferation, (iii) modulation of apoptosis, and/or (iv) treatments of cell proliferation related disorders (e.g., administration alone or co- administration with another molecule). In particular, the compounds of Formula (II) and (Ha) can be used to modulate C 2 activity, in vitro or in vivo, and to treat disorders associated with excessive or undesirable levels of CK2 activity, including cancers, certain inflammatory disorders, vascular disorders, certain skeletal and muscle disorders, and infections such as protozoal parasite infections and some viral infections.

Definitions:

[0054] The terms "a" and "an" do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. The terms "a" and "an" are used interchangeable with "one or more" or "at least one". The term "or" or "and/or" is used as a function word to indicate that two words or expressions are to be taken together or individually. The terms "comprising", "having", "including", and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to"). The endpoints of all ranges directed to the same component or property are inclusive and independently combinable.

[0055] The terms "compound(s) of the invention", "these compounds", "such

compound(s)", "the compound(s)", and "the present eompound(s)" refer to compounds encompassed by structural formulae disclosed herein, e.g., Formula (I), (II), (IF), (Ila), (Ila'), (lib), (lib'), (lie), (II-Th), and (II-Th'), includes any specific compounds within these formulae whose structure is disclosed herein. Compounds may be identified either by their chemical structure and/or chemical name. When the chemical structure and chemical name conflict, the chemical structure is determinative of the identity of the compound. Furthermore, the present compounds can modulate, i.e., inhibit or enhance, the biological activity of a CK2 protein, a Pirn protein or both, and thereby is also referred to herein as a "modulator(s)" or "C 2 and/or Pirn modulator(s)". Compounds of Formula (I), (II), (IF), (Ila), (Ila'), (lib), (lib'), (He), (II-Th), and (II-Th'), including any specific compounds, i.e., species, described herein are exemplar}' "modulators".

[0056] The compounds described herein may contain one or more chiral centers and/or double bonds and therefore, may exist as stereoisomers, such as double-bond isomers (i.e., geometric isomers such as E and Z), enantiomers or diastereomers. The invention includes each of the isolated stereoisomer! c forms (such as the enatitiomerically pure isomers, the E and Z isomers, and etc.) as well as mixtures of stereoisomers in varying degrees of chiral purity or percetange of E and Z, including raceraic mixtures, mixtures of diastereomers, and mixtures of E and Z isomers. Accordingly, the chemical structures depicted herein encompass all possible enan tiomers and stereoisomers of the illustrated compounds including the stereoisomerically pure form (e.g., geometrically pure, enantiomerically pure or diastereomerically pure) and enantiomeric and stereoisomeric mixtures. Enantiomeric and stereoisomers mixtures can be resolved into their component enantiomers or stereoisomers using separation techniques or chiral synthesis techniques well known to the skilled artisan. The invention includes each of the isolated stereoisomeric forms as well as mixtures of stereoisomers in varying degrees of chiral purity, including racemic mixtures. It also encompasses the various diastereomers. Other structures may appear to depict a specific isomer, but that is merely for convenience, and is not intended to limit the invention to the depicted olefin isomer. When the chemical name does not specify the isomeric form of the compound, it denotes any one of the possible isomeric forms or a mixtures of those isomeric forms of the compound.

[0057] The compounds may also exist in several tautomeric forms, and the depiction herein of one tautomer is for convenience only, and is also understood to encompass other tautomers of the form shown. Accordingly, the chemical structures depicted herein encompass all possible tautomeric forms of the illustrated compounds. The term "tautomer" as used herein refers to isomers that change into one another with great ease so that they can exist together in equilibrium. For example, ketone and enol are two tautomeric forms of one compound. In another example, a substituted 1,2,4-triazole derivative may exist in at least three tautomeric forms as shown below:

R^T1 is H or optionally substituted alkyl, R^T2 is an optionally substituted aryl.

[0058] The descriptions of compounds of the present invention are limited by principles of chemical bonding known to those skilled in the art. Accordingly, where a group may be substituted by one or more of a number of substituents, such substitutions are selected so as to comply with principles of chemical bonding and to gi ve compounds which are not inherently unstable and/or would be known to one of ordinary skill in the art as likely to be unstable under ambient conditions, such as aqueous, neutral, and several known physiological conditions. For example, a heterocycloaikyi or heteroaryl is attached to the remainder of the molecule via a ring heteroatom in compliance with principles of chemical bonding known to those skilled in the art thereby avoiding inherently unstable compounds.

[0059] The compounds of the invention often have ionizable groups so as to be capable of preparation as salts. In that case, wherever reference is made to the compound, it is understood in the art that a pharmaceutically acceptable salt may also be used. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases are well-known in the art, such as hydrochloric, sulphuric, hydrobromic, acetic, lactic, citric, or tartaric acids for forming acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, various amines, and the like for forming basic salts. Methods for preparation of the appropriate salts are well-established in the art. In some cases, the compounds may contain both a acidic and a basic functional group, in which case they may have two ionized groups and yet have no net charge. Standard methods for the preparation of pharmaceutically acceptable salts and their formulations are well known in the art, and are disclosed in various references, including for example, "Remington: The Science and Practice of Pharmacy", A. Gennaro, ed., 20th edition, Lippincott, Williams & Wilkins, Philadelphia, PA.

[0060] "Solvate", as used herein, means a compound formed by solvation (the combination of solvent molecules with molecules or ions of the solute), or an aggrega te that consists of a solute ion or molecule, i.e., a compound of the invention, with one or more solvent molecules. When water is the solvent, the corresponding solvate is "hydrate". Examples of hydrate include, but are not limited to, hemihydrate, monohydrate, dihydrate, trihydrate, hexahydrate, etc. It should be understood by one of ordinary skill in the art that the pharmaceutically acceptable salt, and/or prodrug of the present compound may also exist in a solvate form, The solvate is typically formed via hydration which is either part, of the preparation of the present compound or through natural absorption of moisture by the anhydrous compound of the present invention.

[0061] The term "ester" means any ester of a present compound in which any of the -COOH functions of the molecule is replaced by a -COOR function, in which the I moiety of the ester is any carbon-containing group which forms a stable ester moiety, including but not limited to alkyS, aikenyl, alkynyl, eycloalkyl, cyeloalkyialkyl, aryS, aiylalkyi heterocyclyl,

heterocyclylalkyl and substituted derivatives thereof. The hydrolysabie esters of the present compounds are the compounds whose carboxyls are present in the form of hy droly sabie ester groups. That is, these esters are pharmaceutically acceptable and can be hydrolyzed to the corresponding carboxyl acid in vivo. These esters may be conventional ones, including lower alkanoyloxyalkyl esters, e.g. pivaloyloxymethyl and 1-pivaloyloxyethyl esters; lower

alkoxycarbonylalkyl esters, e.g., meihoxycarbonyioxyrneihyl, 1 -ethoxycarbonyloxyethyl, and 1- isopropylcarbonyloxyethyl esters; lower alkoxymethyl esters, e.g., methoxymethyl esters, iactonyl esters, benzofuran keto esters, thiobenzofuran keto esters; lower alkanoylamiiiomethyl esters, e.g., acetylaminoraethyl esters. Other esters can also be used, such as benzyl esters and cyaiio methyl esters. Other examples of these esters include: (2,2-diniethyl-i- oxypropyloxy)methyl esters; (1 RS)-1 -acetoxyethyl esters, 2-[(2-methylpropyloxy)carbonyI]-2- pentenyl esters, l-[[(l-methylethoxy)carbonyl]- oxyjethyl esters; isopropyloxycarbonyloxyethyl esters, (5-methyl-2-oxo-l ,3- dioxole-4-yl) methyl esters, l-[[(cyclohexyloxy)carbonyl]oxy]ethyl esters; 3,3-dimethyl-2-oxobutyl esters. It is obvious to those skilled in the art that hydrolysabie esters of the compounds of the present invention can be formed at free carboxyls of said compounds by using conventional methods. Representative esters include pivaloyloxymethyl esters, isopropyloxycarbonyloxyethyl esters and (5-methyl-2-oxo- 1 ,3-dioxole-4-yl)methyl esters.

[0062] The term "prodrug" refers to a precursor of a pharmaceutically active compound wherein the precursor itself may or may not be pharmaceutically active but, upon administration, will be converted, either metabolically or otherwise, into the pharmaceutically active compound or drug of interest. For example, prodrug can be an ester, ether, or amide form of a

pharmaceutically active compound. Various types of prodrug have been prepared and discl osed for a variety of pharmaceuticals. See, for example, Bundgaard, H. and Moss, J,, J. Pharm. Sci. 78: 122-126 (1989), Thus, one of ordinary skill in the art knows how to prepare these prodrugs with commonly employed techniques of organic synthesis. [0063] "Protecting group" refers to a grouping of atoms that when attached to a reactive functional group in a molecule masks, reduces or prevents reactivity of the functional group. Exampl es of protecting groups can be found in Green et aL, "Protective Groups in Organic Chemistry", (Wiley, 2^nd ed. 1991 ) and Harrison et aL, "Compendium of Synthetic Organic Methods", Vols, 1-8 (John Wiley and Sons, 1971-1996). Representative amino protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonvi ("CBZ"), teri-butoxycarbonyl ("Boc"), trimethylsilyl ("TMS"), 2-ttimethylsilyl-ethanesulfonyl ("SES"), trityl and substituted trityl groups, aliyloxycarbonyl, 9-fluorenylmethyloxycarbonyi ("FMOC"), nitro-veratryloxycarbonyl (" VOC") and the like. Representative hydroxy protecting groups include, but are not limited to, those where the hydroxy group is either acylated or alkylated such as benzyl, and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers, iriaikyisilyl ethers and allyl ethers.

[0064] As used herein, "pharmaceutically acceptable" means suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use within the scope of sound medical judgment.

[0065] "Excipient" refers to a diluent, adjuvant, vehicle, or carrier with which a compound is administered.

[0066] An "effective amount" or "therapeutically effective amount" is the quantity of the present compound in which a beneficial outcome is achieved when the compound is

administered to a patient or al ternatively, the quantity of compound that possesses a desired activity in vivo or in vitro. In the case of proliferative disorders, a beneficial clinical outcome includes reduction in the extent or severity of the symptoms associated with the disease or disorder and/or an increase in the longevity and/or quality of life of the patient compared with the absence of the treatment. For example, for a subject with cancer, a "beneficial clinical outcome" includes a reduction in tumor mass, a reduction in the rate of tumor growth, a reduction in metastasis, a reduction in the severity of the symptoms associated with the cancer and/or an increase in the longevity of the subject compared with the absence of the treatment. The precise amount of compound administered to a subject will depend on the type and severity of the disease or condition and on the characteristics of the patient, such as general health, age, sex, body weight and tolerance to drugs. It will also depend on the degree, severity and type of proliferative disorder. The skilled artisan will be able to determine appropriate dosages depending on these and other factors. [0067] As used herein, the terms "alkyl," "alkenyl" and "alkynyl" include straight-chain, branched-chain and cyclic monovalent hydrocarbyl radicals, and combinations of these, which contain only C and H when they are unsubstituted. Examples include methyl, ethyl, isobutyl, cyclohexyl, cyclopentylethyl, 2-propenyl, 3-butynyl, and the like. The total number of carbon atoms in each such group is sometimes described herein, e.g., when the group can contain up to ten carbon atoms it can be represented as 1 -1 OC or as C 1 -C 10 or C 1 - \ 0. When heieroatoms ( N, O and S typically) are allowed to replace carbon atoms as in heteroalkyl groups, for example, the numbers describing the group, though still written as e.g. C1-C6, represent the sum of the number of carbon atoms in the group plus the number of such heieroatoms that are included as replacements for carbon atoms in the backbone of the ring or chain being described. Where a ring is included, it is understood that the group contains at least three carbon atoms as a 3- membered ring is the smallest size for a ring.

[0068] Typically, the alkyl, alkenyl and alkynyl substituents of the invention contain 1-lOC (alkyl) or 2- IOC (alkenyl or alkynyl), or 3- IOC when a ring is included. Preferably they contain 1 -8C (alkyl) or 2-8C (alkenyl or alkynyl) or 3-8C when a ring is included. Sometimes they contain 1-4C (alkyl) or 2-4C (alkenyl or alkynyl). A single group can include more than one type of multiple bond, or more than one multiple bond; such groups are included within the definition of the term "alkenyl" when they contam at least one carbon-carbon double bond, and are included within the term "alkynyl" when they contain at least one carbon-carbon triple bond; provided, however, that the presence of multiple bonds does not produce an aromatic ring.

[0069] Alkyl, alkenyl and alkynyl groups are often optionally substituted to the extent that such substitution makes sense chemically.

[0070] "Optionally substituted" as used herein indicates that the particular group or groups being described may have no non-hydrogen substituents, or the group or groups may have one or more non-hydrogen substituents. If not otherwise specified, the total number of such substituents that may be present is equal to the number of H atoms present on the unsubstituted form of the group being described. Where an optional substituent is a ttached via a double bond, such as a carbonyl oxygen (^::Ό), the group takes up two available valences, so the total number of substituents that may be included is reduced according to the number of available valences.

[0071] "Substituted," when used to modify a specified group or radical, means that one or more hydrogen atoms of the specified group or radical are each, independently of one another, replaced with the same or different substituent(s). [0072] Substitueni groups useful for substituting saturated carbon atoms in the specified group or radical include, but are not limited to -R^a, halo, -O^", =0, -OR^b, -SR^b, -S^", =S, -NR^CR^C,

N R^1'. X-OR^1'. trihalomethyl, -CF₃, -CN, -OCN, -SCN, -NO, ~NQ₂, =N₂, -N₃, -8(0)₂R^b, -S(0)₂NR^b, -S(0)₂0^", -S(0)₂OR^b, -OS(0)₂R^b, -OS(0)₂0^", -OS(0)₂OR^b, -P(0){0^")₂,

-P(0)(OR^b)(0^"), -P(0)(OR^b)(OR^b), -C(0)R^b, -C(S)R^b, -C(NR^b)R^b, -0(0)0^", -C(0)OR^b, -C(S)OR^b, -C(())NR^CR^C, -C(NR^b)NR^cR^c, -OCf())R^b, -OC(S)R^b, -OC(0)0^", ~OC(0)OR^b, -OC(S)OR^b, -NR^bC(0)R^b, -NR^bC(S)R^b. -NR^bC(0)0^", -NR^bC(0)OR^b, -NR^bC(S)OR^b,

-NR^bC(0)NR^cR^c, -NR C(NR )R^b and -NR^bC(NR^b)NR^cR^c, where R^'' is selected from the group consisting of aikyi, cvcloalkyl, heteroalkyl, cycloheteroaikvi , aryi, arylalkyl, heteroaryl and heteroarylalkyl; each R^b is independently hydrogen or R^a; and each R^c is independently R^b or alternatively, the two R^cs may be taken together with the nitrogen atom to which they are bonded form a 4-, 5-, 6- or 7-membered cycloheteroaikvi which may optionally include from 1 to 4 of the same or different additional heteroatoms selected from the group consisting of O, N and S. As specific examples, -NR^CR^C is meant to include -NH₂, -NH-alkyl, N-pyrrolidinyl and N-morpholinyS. As another specific example, a substituted a!kyl is meant to include --alkylene- O-alkyl, -alkylene-heteroaryl, -alkylene-cycloheteroalkyl, -alkylene-C(0)OR°, -alkylene- C(0)NR^bR^b, and -CH₂-CH₂-C(0)-CH₃. The one or more substitueni groups, taken together with the atoms to which they are bonded, may form a cyclic ring including cycloalkvl and cycloheteroaikvi ,

[0073] Similarly, substitueni groups useful for substituting unsaturated carbon atoms in the specified group or radical include, but are not limited to, -R^a, halo, -O^", -OR^b, -SR°, -S^", -NR R\ trihalomethyl, -CF₃, -CN, -OCN, -SCN, -NO, -N0₂, -N₃, -S(0)₂R^b, -S(0)₂0^", -S(0)₂OR^b, -OS(0)₂R^b, -OS(0)₂0^", -OS(0)₂OR^b, -P(0)(0^")₂, -P(0)(OR^b)(0^"), -P(0)(OR^b)(OR^b), -C(0)R^b, -C(S)R^b, -C(NR^b)R^b, -C(())0^", -C(0)OR^b, -C(S)OR^b, -C(0)NR^cR^c, -C(NR^b)NR^cR^c, -OC(0)R^b, -OC(S)R^b, -OC(0)0^", -OC(0)OR^b, -OC(S)OR^b, -NR^bC(0)R^b, -NR^bC(S)R^b, -NR^bC(0)0^",

-NR^bC(0)OR^b, - R^bC(S)OR^b, ~NR^bC(0)NR^cR^c, -NR^bC(NR^b)R^b and -NR^bC(NR^b)NR^cR^c, where R , R^b and R^c are as previously defined,

[0074] Substitueni groups useful for substituting nitrogen atoms in heteroalkyl and cycloheteroaikvi groups include, but are not limited to, -R^a, -O^", -OR⁰, -SR°, -S^", -NR^CR^C, trihalomethyl, -CF₃, -CN, -NO, -NQ₂, -S(0)₂R^b, -S(0)₂0^", -S(0)₂OR^b, -OS(0)₂R^b, -OS(0)₂0; -OS(0)₂OR^b, -P(0)(0^")2, -P(0)(OR^b)(01, -P(0)(OR^b)(OR^b), -C(0)R^b, -C(S)R^b, -C(NR^b)R^b, -C(0)OR^b, -C(S)OR^b, -C(0)NR^cR^c, -C(NR^b)N R^cR^c, -OC(0)R^b, -OC(S)R^b, -OC(0)OR^b, -OC(S)OR^b, -NR^bC(0)R^b, -NR^bC(S)R^b, -NR^bC(0)OR^b, -NR^bC(S)OR^b, ~NR^bC(0)NR^cR^c, -NR^bC(NR^b)R^b and -NR^bC(NR^b)NR^cR^c, where R^a, R^b and R^c are as previously defined.

[0075] Alky], alkenyl and alkynyl groups can alternatively or in addition be substituted by C1-C8 acyl, C2-C8 heteroacyl, C6-C10 aryi, C3-C8 eycloalkyl, C3-C8 heterocyclyl, or C5-C10 heteroaryl, each of which can be substituted by one or more R, halo, =0, =N-CN, =N-OR, =NR, OR, NR₂, SR, S0₂R, SO -XR -, NRS0₂R, NRCONR₂, NRCSNR₂, NRC(=NR)NR₂, NRCOOR, NRCOR, CN, C≡CR, COOR, CONR₂, OOCR, COR, and N0₂, wherein each R is independently H, C1 -C8 alkyl, C2-C8 heteroalkyl, C1-C8 acyl, C2-C8 heteroacyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C3-C8 heterocyclyl, C4-C10

heterocyclyl alkyl, C6-C10 aryl, or C5-C10 heteroaryl, and each R is optionally substituted with one or more (typically up to three) halo, =0, =N-CN, =N-OR', =NR', OR', NR.'₂, SR', S0₂R', S0₂NR'₂, NR'S0₂R', NR'CONR'₂, NR'CSNR'?, NR'C(=NR') R'₂, NR'COOR', NR'COR', CN, C≡CR', COOR', CONRS, OOCR', COR', and N0₂, wherem each R' is independently H, C1-C8 alkyl C2-C8 heteroalkyl, C1-C8 acyl, C3-C8 heterocyclyl, C2-C8 heteroacyl, C6-C10 aryl or C5-C10 heteroaryl.

[0076] Where any of these substituents contains two R or R' groups on the same or adjacent atoms (e.g., ~NR₂, or -^'NR-C(O)R), the two R or R' groups can optionally be taken together with the atom(s) in the substituent group to which they are attached to form a ring having 5-8 ring members, which can include another heteroatom as a ring member (N, O or S) and can be substituted with one or more halo, =0, =N-CN, =N-OR, =NR, OR, NR₂, SR, S0₂R, S0₂NR₂, NRS0₂R, NRCONR2, NRCS^'NR-2, NR.C(^::::NR)^'NR-2, NRCOOR, NRCOR, CN, OCR, COOR, CONR₂, OOCR, COR, and N0₂, wherem each R is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C1-C8 acyl, C2-C8 heteroacyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyl, C3-C8 heterocyclyl, C4-C10 heterocyclylalkyl, C6-C10 aryl, or C5-C10 heteroaryl, and each R is optionally substituted with halo, =0, =N-CN, =N-QR', =NR', OR', NR'₂, SR', S0₂R', SOAR^" -- NR'S0₂R\ NR'C0NR'₂, NR'CSNR'₂, NR'C(==NR')NR'₂, NR'COOR', NR'COR', CN, C≡CR', COOR', CONR'₂, OOCR', COR', and N0₂, wherein each R^" is independently H, C1 -C8 alkyl, C2-C8 heteroalkyl, C1-C8 acyl, C3-C8 heterocyclyl, C2-C8 heteroacyl, C6-C10 aryl or C5-C10 heteroatyl, and each of the substitutable groups on R ' can be substituted with one or more (e.g., up to three) halo, piperidinyl, pyrrolidinyl, piperazinyl, morphoiinyl, CN, C1-C4 alkoxy, OH, OAc, NH₂, C1-C4 alkyl amine, di(Cl-C4 alkyl)amine, M l Ac. NHCOOMe, NHCOOEt, NHCOOtBu, NHS0₂Me, S e, S0₂Me, S0₂N1¾, S0₂NMe₂, COOH, CONH₂, .·( )( )Me. COOEt, CONHMe, or CONMe₂. [0077] "Acetylene" substituents are 2- IOC alkynyl groups that contain at least one carbon- carbon triple bond and are optionally substituted with the groups described herein as suitable for alkyl groups; in some embodiments, the alkynyl groups are of the formula ~C≡C~R^a, wherein R^a is H or C1 -C8 alkyl, C2-C8 heteroalkyl, C2-C8 aikenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl, C2-C8 heteroalkynyi, C1-C8 acyl, C2-C8 heteroacyi, C6-C10 and, C5-C10 heteroaryi, C7-C12 arylalkyl, or C6-C12 heteroarylalkyl.

[0078] Each R^d group is optionally substituted with one or more substituents selected from halo, ⁽ X =N-CN, X~OR\ XR^". OR', NR'₂, SR', S0₂R', SC>_::XR\_:. NR'S()₂R\ NR'CONR'₂, NR'CSNRS, NR.' C(=NR' )NR' ₂, NR'COOR', NR'COR', CN, COOR', CONR'₂, OOCR', COR.', and N0₂, wherein each R' is independently H, C1-C6 alkyl, C2-C6 heteroalkyl, C1 -C6 acyl, C2-C6 heteroacyi, C6-C 10 aryl, C5-C10 heteroaryi, C7-12 arylalkyl, or C6-12

heteroarylalkyl, each of which is optionally substituted with one or more groups selected from halo, CN, C 1-C4 alkyl, C2-C4 heteroalkyl, C1-C6 acyl, C 1-C6 heteroacyi, C1-C4 alkoxy, Cl - C4 alkylamino, di(Cl-C4 aikyi)amino, hydroxy, amino, and =0; and wherein two R' can be linked to form a 3-7 membered ring optionally containing up to three heteroatoms selected from N, O and S. In some embodiments, R³ of -C≡C-R^a is H or Me.

[0079] "Heteroalkyl", "heteroalkenyl", and "heteroalkynyi" and the like are defined similarly to the corresponding hydrocarbyl (alkyl, aikenyl and alkynyl) groups, but the 'hetero' terms refer to groups that con tain 1-3 O, S or N heteroatoms or combinations thereof within the backbone residue; thus at least one carbon atom of a corresponding alkyl, aikenyl, or alkynyl group is replaced by one of the specified heteroatoms to form, respectively, a heteroalkyl, heteroalkenyl, or heteroalkynyi group. The typical and preferred sizes for heteroforms of alkyl, aikenyl and alkynyl groups are generally the same as for the corresponding hydrocarbyl groups, and the substituents that may be present on the heteroforms are the same as those described above for the hydrocarbyl groups. For reasons of chemical stability, it is also understood that, unless otherwise specified, such groups do not include more than two contiguous heteroatoms except where an oxo group is present on N or S as in a nitro or sulfonyl group.

[0080] While "alkyl" as used herein includes eyeloalkyl and cycloalkylalkyl groups, the term "eyeloalkyl" may be used herein to describe a carbocyclic non-aromatic group that is connected via a ring carbon atom, and "cycloalkylalkyl" may be used to describe a carbocyclic non-aromatic group that is connected to the molecule through an alkyl linker,

[0081] Similarly, "heterocyclyl" may be used to describe a non-aromatic cyclic group that contains at least one heteroatom (typically selected from N, O and S) as a ring member and that is connected to the molecule via a ring atom, which may he C (carbon-linked) or N (nitrogen- linked); and "heterocyclylalkyl" may be used to describe such a group that is connected to another molecule through a linker. The heterocyclyl can be fully saturated or partially saturated, but non-aromatic. The sizes and substituents that are suitable for the cycloalkyl,

cycloalkylalkyl, heterocyclyl, and heterocyclylalkyl groups are the same as those described above for alky! groups. The heterocyclyl groups typically contain 1, 2 or 3 heteroatoms, selected from N, O and S as ring members: and the N or S can be substituted with the groups commonly found on these atoms in heterocyclic systems. As used herein, these terms also include rings that contain a double bond or two, as long as the ring that is attached is not aromatic, The substituted cycloalkyl and heterocyclyl groups also include cycloalkyl or heterocyclic rings fused to an aromatic ring or heteroaromatic ring, provided the point of attachment of the group is to the cycloalkyl or heterocyclyl ring rather than to the aromatic / heteroaromatic ring.

[0082] Like alkyl groups, the cycloalkyl and heterocyclyl groups described herein can be substituted to the extent permitted by their valence and stability considerations, which are well understood by those of skill in the art. Substituents for the cycloalkyl and heterocyclyl rings or ring systems include those described herein as suitable for placement on alkyl groups.

[0083] As used herein, "acyl" encompasses groups comprising an alkyl, aikenyl, alkynyl, aryl or arylalkyl radical attached at one of the two available valence positions of a carbonyl carbon atom, and heteroacyl refers to the corresponding groups wherein at least one carbon other than the carbonyl carbon has been replaced by a heteroatom chosen from N, O and S. Thus heteroacyl includes, for example, -C(=0)OR and -C(=0)NR₂ as well as -C(=0)-heteroaryL

[0084] Acyl and heteroacyl groups are bonded to any group or molecule to which they are attached through the open valence of the carbonyl carbon atom. Typically, they are C1-C8 acyl groups, which include forniyl, acetyl, pivaloyl, and benzoyl, and C2-C8 heteroacyl groups, which include methoxyacetyl, ethoxycarbonyl, and 4-pyridmoyl. The hydrocarbyl groups, aryl groups, and hetero forms of such groups that comprise an acyl or heteroacyl group can be substituted with the substituents described herein as generally suitable substituents for each of the corresponding component of the acyl or heteroacyl group.

[0085] "Aromatic" moiety or "aryl" moiety refers to a monocyclic or fused bicyclic moiety having the w^rell-known characteristics of aromaticity; examples include phenyl and naphthyi. Similarly, "heteroaromatic" and "heteroaryl" refer to such monocyclic or fused bicyclic ring systems which contain as ring members one or more heteroatoms selected from (), S and N. The inclusion of a heieroatom pennits aromaticity in 5-membered rings as well as 6-membered rings, Typical heteroaromatic systems include monocyclic C5-C6 aromatic groups such as pyridyl, pyrimidyl, pyrazinyl, thienyl, furanyl, pyrroiyl, pyrazolyl, thiazolyl, oxazolyl, triazolyl, triazinyl, tetrazolyl, tetraziny!, and imidazolyl and the fused bicyclic moieties formed by fusing one of these monocyclic groups with a phenyl ring or with any of the heteroaromatic monocyclic groups to form a C8-C10 bicyclic group such as indolyl, benzimidazolyl, indazolyl,

beiizotriazolyl, isoquinoiyl, quinolyl, benzothiazolyl, benzofuranyl, pyrazolopyridyl,

quinazoliny!, quinoxalinyS, cinnolinyl, and the like. Any monocyclic or fused ring bicyclic system which has the characteristics of aromaticity in terms of electron distribution throughout the ring system is included in this definition. It also includes bicyclic groups where at least the ring which is directly attached to the remainder of the molecule has the characteristics of aromaticity. Typically, the ring systems contain 5-12 ring member atoms and up to four heteroatoms selected from , O and S. Frequently, the monocyclic heteroaryls contain 5-6 ring members and up to three such heteroatoms, and the bicyclic heteroaryls contain 8-10 ring members and up to four such heteroatoms. The number and placement of heteroatoms in such rings is in accordance with the well-known limitations of aromaticity and stability, where stability requires the heteroaromatic group to be stable enough to be exposed to water without rapid degradation.

[0086] Aryl and heteroaryl moieties may be substituted with a variety of substituents including C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C5-C 12 aryl, C1-C8 acyl, and

hetero forms of these, each of which can itself be further substituted; other substituents for aryl and heteroaryl moieties include halo, OR, NR₂, SR. SO,R. 80₂NR₂, NRS0₂R, NRCONR₂, NRC8NR-2, NRC(=NR)NR₂, NRCOOR, NRCOR, CN, V CR. COOR, CONR¾ OOCR, COR, and NO?, wherein each R is independently H, C1-C8 alkyl, C2-C8 heteroalkyl, C2-C8 alkenyl, C2-C8 heteroalkenyl, C2-C8 alkynyl C2-C8 heteroalkynyl, C3-C8 heterocyclyl C4-C10 heterocyclylalkyl, C6-C10 aryl, C5-C10 heteroaryl, C7-C12 arylalkyl, or C6-C12

heteroarylalkyl, and each R is optionally substituted as described above for alkyl groups. The substituent groups on an aryl or heteroaryl group may of course be further substituted with the groups described herein as suitable for each type of such substituents or for each component of the substituent, Thus, for example, an arylalkyl substituent may be substituted on the aryl portion with substituents described herein as typical for aryl groups, and it may be further substituted on the alkyl portion with substituents described herein as typical or suitable for alkyl groups. Where a substituent group contains two R groups on the same or adjacent atoms (e.g., - NR.2, or -NR-C(O)R), the two R groups can optionally be taken together with the atom(s) in the substituent group to which the are attached to form a ring having 5-8 ring members, which can be substituted as allowed for the R. itself, and can contain an additional heteroatom (N, O or S) as a ring member.

[0087] Similarly, "arylalkyl" and "heteroarylalkyl" refer to aromatic and heteroaromatic ring systems which are bonded to their attachment point through a linking group such as an alkylene, including substituted or unsubstituted, saturated or unsaturated, cyclic or acyclic linkers, Typically the linker is C1 -C8 alky! or a hetero form thereof. These linkers may also include a carbonyl group, thus making them able to provide substituents as an acyl or heteroacyl moiety. An aryl or heteroaryl ring in an arylalkyl or heteroarylalkyl group may be substituted with the same substituents described above for aryl groups. Preferably, an arylalkyl group includes a phenyl ring optionally substituted with the groups defined above for aryl groups and a C1-C4 alkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl groups or heteroalkvl groups, where the alkyl or heteroalkvl groups can optionally cyciize to form a ring such as cyclopropane, dioxo!ane, or oxacyc!opentane. Similarly, a heteroarylalkyl group preferably includes a C5-C6 monocyclic heteroaryl group that is optionally substituted with the groups described above as substituents typical on aryl groups and a C1-C4 alkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl groups or heteroalkvl groups, or it includes an optionally substituted phenyl ring or C5-C6 monocyclic heteroaryl and a C1-C4 heteroalkylene that is unsubstituted or is substituted with one or two C1-C4 alkyl or heteroalkvl groups, where the alkyl or heteroalkvl groups can optionally cyciize to form a ring such as cyclopropane, dioxolane, or oxacyclopentane.

[0088] Where an arylalkyl or heteroarylalkyl group is described as optionally substituted, the substituents may be on either the alkyl or heteroa!ky! portion or on the aryl or heteroaryl portion of the group. The substituents optionally present on the alkyl or heteroalkvl portion are the same as those described above for alkyl groups generally; the substituents optionally present on the aryl or heteroaryl portion are the same as those described above for aryl groups generally.

[0089] "Arylalkyl" groups as used herein are hydrocarbyl groups if they are unsubstituted, and are described by the total number of carbon atoms in the ring and alkylene or similar linker. Thus a benzyl group is a C7-arylalkyl group, and phenylethyl is a C8-arylalkyl.

[0090] "Heteroarylalkyl" as described above refers to a moiety comprising an aryl group that is attached through a linking group, and differs from "arylalkyl" in that at least one ring atom of the aryl moiety or one atom in the linking group is a heteroatom selected from N, O and S. The heteroarylalkyl groups are described herein according to the total number of atoms in the ring and linker combined, and they include aryl groups linked through a heteroalkyl linker;

heteroaryl groups linked through a hydrocarbyi linker such as an alkylene; and heteroaryl groups linked through a heteroalkyl linker. Thus, for example, C7-heteroarylalkyl would include pyridylmethyl, phenoxy, and N-pyrrolylmethoxy.

[0091 ] "Alkylene" as used herein refers to a divalent hydrocarbyi group; because it is divalent, it can link two other groups together, Typically it refers to -(CH;:>)_n- where n is 1-8 and preferably n is 1 -4, though where specified, an alkylene can also be substituted by other groups, and can be of other lengths, and the open valences need not be at opposite ends of a chain. Thus -CH(Me)- and -C(Me)₂- may also be referred to as alkylenes, as can a cyclic group such as eyclopropan-l ,l-diyl. Where an alkylene group is substituted, the substituents include those typically present on alky] groups as described herein,

[0092] In general, any alkyl, alkenyl, alkynyl, acyl, or aryl or arylalkyl group or any heteroform of one of these groups that is contained in a substituent may itself optionally be substituted by additional substituents. The nature of these substituents is similar to those recited with regard to the primary substituents themselves if the substituents are not otherwise described. Thus, where an embodiment of, for example, R^x is alkyl, this alkyl may optionally be substituted by the remaining substituents listed as embodiments for R where this makes chemical sense, and where this does not undermine the size limit provided for the alkyl per se; e.g., alkyl substituted by alkyl or by alkenyl would simply extend the upper limit of carbon atoms for these embodiments, and is not included. However, alkyl substituted by aryl, amino, alkoxy, =0, and the like would be included within the scope of the invention, and the atoms of these substituent groups are not counted in the number used to describe the alkyl, alkenyl, etc, group that is being described. Where no number of substituents is specified, each such alkyl, alkenyl, alkynyl, acyl, or aryl group may be substituted with a number of substituents according to its available valences; in particular, any of these groups may be substituted with fluorine atoms at any or all of its a vailable valences, for example.

[0093J "Heteroform" as used herein refers to a derivative of a group such as an alkyl, aryl, or acyl, wherein at least one carbon atom of the designated carbocyclic group has been replaced by a heteroatom selected from N, O and S, Thus the he ero forms of alkyl, alkenyl, alkynyl , acyl, aryl, and arylalkyl are heteroalkyl, heteroalkenyl, heteroalkynyl, heteroacyl, heteroaryl, and heteroarylalkyl, respectively. It is understood that no more than two N, O or S atoms are ordinarily connected sequentially, except where an oxo group is attached to N or S to form a nitro or sulfonyi group.

[0094] "Halo", as used herein includes fluoro, chloro, bromo and iodo. Fluoro and chloro are often preferred.

[0095] "Amino" as used herein refers to NH₂, but where an amino is described as

"substituted" or "optionally substituted", the term includes R'R" wherein each R' and R" is independently H, or is an alkyl, alkenyl, alkynyl, acyl, aiyl, or aryla.lkyl group or a heteroform of one of these groups, and each of the alkyl, alkenyl, alkynyl, acyl, aryl, or aryl alkyl groups or heteroforms of one of these groups is optionally substituted with the substituents described herein as suitable for the corresponding group. The term also includes forms wherein R' and R" are taken together with the N to which they are attached to form a 3-8 membered ring which may be saturated, unsaturated or aromatic and which contains 1 -3 heteroaioms independently selected from N, O and S as ring members, and which is optionally substituted with the substituents described as suitable for alkyl groups or, if NR'R" is an aromatic group, it is optionally substituted with the substituents described as typical for heteroaryl groups.

[0096] As used herein, the term "carbocycle", "carbocyclyi", or "carbocyclic" refers to a cyclic ring containing only carbon atoms in the ring, whereas the term "heterocycle" or

"heterocyclic" refers to a ring comprising a heteroatom. The carbocyclyi can be fully saturated or partially saturated, but non-aromatic. For example, the carbocyclyi encompasses cycloalkyl. The carbocyclic and heterocyclic structures encompass compounds having monocyclic, bicyclic or multiple ring systems; and such systems may mix aromatic, heterocyclic, and carbocyclic rings. Mixed ring systems are described according to the ring that is attached to the rest of the compound being described; for example, where W represents 1 ,2,3 ,4-tetrahydronaphth- 1 -yl, the group would be encompassed by an optionally substituted cycloalkyl or carbocyclic group, while the group l,2,3,4~tetrahydronaphth-6-yl would be included within optionally substituted aromatic groups.

[0097] As used herein, the term "heteroatom" refers to any atom that is not carbon or hydrogen, such as nitrogen, oxygen or sulfur, When it is part of the backbone or skeleton of a chain or ring, a heteroatom must be at least divalent, and will typically be selected from N, O, P, and S.

[0098] Illustrative examples of heterocycles and heteroaryls include but are not limited to tetrahydrofuran, 1 ,3-dioxolane, 2,3-dihydrofuran, pyran, tetrahydropyran, benzofuran, isobenzofuran, 1,3-dihydro-isobenzofuran, isoxazole, 4,5-dihydroisoxazole, piperidine, pyrrolidine, pyrrolidin-2-one, pyrrole, pyridine, pyrimidine, octahydro-pyrrolo[3,4 bjpyridine, piperazine, pyrazine, niorpholine, thlomorpholme, imidazole, imidazolidine 2,4-dione, 1,3- dihydrobenzimidazol-2-one, indole, thiazole, henzothiazole, thiadiazole, thiophene, tetrahydro thiophene 1,1. -dioxide, diazepine, triazole, guanidine, diazabicycio[2.2.1 ]heptane, 2,5- diazabicyclo[2,2. Ijheptane, 2,3 ,4_>4a,9,9a-hexahydro- 1 Η-β-carboline, oxirane, oxetane, tetrahydropyran, dioxane, lactones, aziridine, azetidine, piperidine, lactams, and may also encompass heteroar ls. Other illustrative examples of heteroaryls include but are not limited to furan, pyrrole, pyridine, pyrimidine, imidazole, benzimidazole and triazole.

Embodiments of the Compounds:

In one embodiment, the compounds of the invention have the general formula (I):

wherein the bicyclic ring system containing Z¹ to Z^' is aromatic;

one of Z⁵ and Z^"' is C, the other of Z' and Z² is N;

Z³ and Z⁴ are independently CR³ or N,

where R³ can be H or R¹;

R¹ is H, halo, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C1-C4 alkoxy, or -NR'R⁸, where R^' and R ^' are each independently selected from H, optionally substituted CI -CIO alkyl, optionally substituted aryl, optionally substituted arylalkvl, optionally substituted heteroaryl, and optionally substituted heteroaxylalkyl,

or R' and R taken together with the N of -NR R^l form an optionally substituted 5-8 menibered ring that optionally contains an additional heteroatom selected from N, O and S as a ring member:

R ^_ is H, halo, CN, or an optionally substituted group selected from C1-C4 alkyl, C2-C4 alkenyl, and C2-C4 alkynyl; R^J and R⁴ are each independently selected from 1-1 and optionally substituted CI -CI O alkvl;

X is NR.⁶, O, or S, where R⁶ is H or an optionally substituted group selected from C1-C4 alkyl, C2-C4 alkenyl, and C2-C4 alkynyl;

Y is O or S;

W is H, optionally substituted aryl, optionally substituted heteroary!, or -NR⁹R^l°, -OR⁹, S(0)„R^y, optionally substituted carbon-linked heterocyclyl, optionally substituted C3-C8 cycloalkyl, or CR⁹R^{! 0}R^U,

wherein n is 0, 1 or 2,

R⁹ and R^l0 are each independently selected from 1-1, optionally substituted CI -CI O alkyl, optionally substituted aryl, optionally substituted ary!alkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, and optionally substituted heterocyclyl,

or R⁹ and R¹⁰ taken together with the N of -NR R^{f 0} form an optionally substituted 5-8 membered ring that optionally contains an additional heteroatom selected from N, O and S as a ring member, and

R¹¹ is selected from H, optionally substituted CI -CIO alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;

or pharmaceutically acceptable salts, solvates, and/or prodrugs of these compounds, [00100] The compounds of the invention are characterized by a bicyclic aromatic heterocycl ic ring system containing two or more nitrogen atoms: one N atom is shown, and one of Z¹ and 7/ is also N. In certain embodiments of interest, Z' is N and Έ^' is C; in other embodiments, Z' is C and Z is N.

[001 1] Optionally, Z' and/or Z⁴ can also be N. In certain embodiments, they are both C; in other embodiments Z³ is N and Z⁴ is C; and in other embodiments Z^" is N and Z³ is C; while in other embodiments, I and Z^ are both N.

[00102] In addition, the compounds of the invention contain another heterocyclic group linked to the bicyclic group, and the additional heterocyclic group contains an amide linkage within the ring, plus an additional carbonyl or thiocarbonyl (C=0 or C=S). The additional heterocyclic group is linked to the bicyclic group through an exocyclic methylene group (an sp² carbon) that is connected to the five-membered ring of the bicyclic group. [00103] This additional heterocyclic group contains X, which can he NR.^", O or S. In certain embodiments, it is NR⁶, and R⁶ is often H or a small alkyl group, such as Me, Preferably, NR° is NH, In other embodiments, X is O. In certain embodiments, X is S,

[00104J This additional heterocyclic group is substituted with =Y; in some embodiments, Y is O and in some embodiments Y is S.

[00105] The additional heterocyclic group also contains NR.", and R.' in this group can be H or a small alkyl such as Me. In some embodiments, it is a substituted alkyl group such as formyl, acetyl, propionyl, benzoyl, and the like. Preferably, R³ is H.

[ 00106] The s ^{^} carbon connecting the two heterocyclic groups is CR^", where R⁴ can be H or a small alkyl; in preferred embodiments, it is H.

[00107] The five-membered ring of the bicyclic group is substituted by R^". This can be H, halo or a small alkyl, such as Me, Et, CF₃, ~CH_?OMe, vinyl, or acetylene. In preferred embodiments, R² is H.

[00108] The six-membered ring of the bicyclic group is substituted by R^l. This can be a variety of groups, including H, halo or an optionally substituted alkyl, amine or alkoxy group. In some embodiments, it is H, halo, or a small alkyl, such as Me, Et, CF₃, -CH₂OMe₅ vinyl, or acetylene. In certain embodiments, R^! is H, halo, Me, NHMe, NMe₂, CF , or CN. In other embodiments, R¹ is -NR'R⁸. In other embodiments, R⁸ is a C3-6 cycloaikyl.

[00109] The six-membered ring of the bicyclic group is also suhstutited by a group W. This can represent a range of different features while retaining the desired protein kinase modulatory activities. In certain embodiments, W is an optionally substituted aryl or heteroaryl group, often selected from phenyl, pyridyl, pyrimidinyi, and pyrazinyl. In particular, it can be an optionally substituted phenyl group, In specific embodiments, W is phenyl substituted with up to two substituents; in certain embodiments, the phenyl group is substituted by at least one group other than H, such as F, CI, Me, CF₃, CN, OMe, COOH, or COOMe, at the ortho or meta position relative to the point at which the phenyl is connected to the bicyclic group.

[00110] Specific embodiments of the substituted phenyl that can be W include 2- flourophenyi, 3~fiuoropheriyl, 3-ca.rboxyphenyl, and 3-(COOMe)-phenyl.

[00111] In other embodiments, W can be a group of the formula -NR⁹Rⁿ', where R⁹ and R'° are as described above. Typically, R⁹ and R¹⁰ are not both H. In certain of these embodiments, R⁹ is H, Me, or an acyl group such as formyl, acetyl, methoxyacetyl, benzoyl, or trifluoroacetyl; such acylated compounds may be active as kinase inhibitors, or they can serve as prodrugs for compounds wherein R⁹ is H. In these embodiments, R³⁰ can be an optionally substituted alkyl group, or an aryl or tieteroaryl group, such as phenyl, pyridinyl, pyrimidinyl, pyrazinyl, and the like, which can be optionally substituted. Suitable optionally substituted alkyl groups include C1-C6 a iky is, e.g., methyl, ethyl, butyl, propyl, isopropyl, t-butyl, llouroethyi, methoxyethyo, isobutyl, and the like, in certain embodiments, the aryl or heteroaryl group is substituted by at least one non-H substituent group. R¹⁰ can also be such an aryl or heteroaryl group that is connected to NR⁹ through a CI ~C4 a Iky 1 en e chain; e.g., it can be imidazolylmethyl, phenylethyl, and the like. In specific embodiments, the aryl is phenyl, and is substituted by at least one non- H substituent, often at the position that is meta or para to the point where the phenyl is connected to the of NR⁹R'°.

[001121 The substituent(s) on this aryl or heteroaryl group ca be halo, C1-C4 alkyl, or C1- C4 alkoxy groups, or aryl or heteroaryl groups such as imidazole, phenyl, pyridyl, pyrazolyl, triazolyi, and the like; or they can be C5-C8 heterocyclic groups such as morpholine, piperidine, piperazine, and the like. In some embodiments, the aryl ring (e.g., phenyl) represented by RIO is substituted with a group of the formula R'2N-(CH2)p-L- , where p is 0-3, L is a bond, O, S, or NR" (R" is H or C1 -C4 alkyl), and each R' is independently H or C1 -C6 alkyl that is optionally substituted, and wherein the two R' groups can optionally cyclize to form a ring, which can include an additional heteroatoni (N, O or 8) as a ring member. Representative examples of this version of R10 include dimethylamino; 4-methylpiperazinyl; 4-morphoiinyl; 4- morpholinomethyl; 4- e-piperazinoethyl; dimethyl aminomethyl; diethylaminomethyl;

dimethylaminoethoxy, and the like.

[00113] Alternatively, R¹⁰ can be an arylalkyl or heteroarylalkyl group, such as an optionally substituted benzyl group. In certain embodiments of Formula I, R^R' is an optionally substituted carbon-linked heterocyclyl.

[00114] Alternati ely, W can be ~NR⁹R¹⁰, where R⁹ and R^{] 0} taken together with N form a ring, which in some embodiments is an optionally substituted 5-8 membered ring that can optionally contain N, O or S as an additional ring member. Exemplary rings mclude piperidine, piperazine, homopiperazine, morpholine, thiomorpholine, pyrrolidine, pyrrolidinone, and the like. In certain embodiments, suhstituents on such rings are Cl-4 alkyl or heteroaryl.

[00115] In certain embodiments of Formula I, W is H.

[00116] In Formula I, X and Y each represent a heteroatom, and they can be the same or they can be different. In some embodiments, Y is O, while X is S or NH or NMe or O; in other embodiments, Y is 8, while X is S, or NH, or NMe or O. Where X is NR.6, R6 can be H, methyl, ethyl, methoxyethyl, and the like; in preferred embodiments, R6 is H or it is Me. [00117] The compounds of the invention include compounds of Formulae I that contain the features specifically described below, or any combination of these features.

[00118] In certain embodiments of Formulae I, Z¹ is N and Z is C.

[00119] in certain embodiments of Formulae I, I is N.

[00120] In certain embodiments of Formulae I, Z^"' is CR⁵,

[00121] in certain embodiments of Formulae I, X is NR^fi or S.

[00122] In certain embodiments of Formulae I, R² is H or Me.

[00123] in certain embodiments of Formulae I, R^J and R⁴ are both H.

[00124] In certain embodiments of Formulae I, R^! is H, Me, halo, OMe, or CF₃.

[00125] In certain embodiments of Formula I, R¹ is -NR R ⁸, wherein R⁸ is C3-6 cycloalkyl.

[00126] In certain embodiments of Formulae I, Y is O.

[00127] In certain embodiments of Formulae I, Y is S.

[00128] In certain embodiments of Formulae I, W is -NH-A, wherein A is optionally substituted phenyl. In alternative embodiments of the above compounds, W is optionally substituted aryl or optionally substituted heteroaryl. In specific embodiments of this type, W can be optionally substituted phenyl. In certain embodiments of Formula I, W is H. In other embodiments of Formula I, W is -N⁹R¹ wherein R¹ is an optionally substituted heterocyclyl.

[00129] In another embodiment, the compounds of Formula (I) have structural Formula (II) or (IF) as shown below (including Ila, Ila', Hb, lib', II-TH, and 11 -^* J ^' I V ). These compounds are typically more selective for CK2, and are highly potent on CK2.

(II) (I F)

wherein:

Z^J and Z⁴ each independently represent N or CR:\ or CH; each R^J is independently selected from halo, ~CN, ~R, -OR, ~S(0)_nR, -COOR, -CONR₂, and -NR₂,

wherein each R is independently selected from H and optionally substituted C1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that optionally contains one or more additional heteroatom selected from N, O and S as a ring member;

R², R³ and R⁴ are each independently selected from H and optionally substituted

C I -C I O alkyl;

X represents O, S, or NR²;

Y is O or S or NR.¹⁰;

where R'° is selected from H, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C.1 -C4 alkoxy, and -NR⁷R⁸,

Z is O or S;

L is a bond, -CR⁷=CR⁸-, -C≡C-, -NR⁷-, -0-, ~S(0)_n-, -(CR⁷RV, -(CR⁷R⁸)_m-NR⁷-, -

(CR⁷RVG-, or -(CR⁷R⁸V-S(0)_n-;

W is optionally substituted CI -C I O alkyl, optionally substituted C I -C I O heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, -NR'R⁸, -OR', -S(0)_NR ' , - CONR 'R⁸, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted C2-C10 alkenyl, optionally substituted C2-C10 alkynyl, or -CR'R⁸R⁹;

where each R' and R⁸ and R⁹ is independently selected from H, optionally substituted C I -C I O alkyl, optionally substituted heteroalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted aryialkyl, optionally substituted heteroaryl, and optionally substituted heteroaiylalkyl;

or R ^' and R , taken together with the carbon atom to which they are attached, form =0 (oxo) or =N-OR⁷ or =N-CN;

or R ' and R , taken together on a single carbon atom or on adjacent connected carbon atoms of (CR'R⁸)_m whether alone or as part of another group, form a 3 to 8 membered carbocyclic ring or heterocyclic ring;

or R^? and R⁸, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 to 10 membered heterocyclic or heteroaryl ring that optionally contains one or more additional heteroatom selected from N, O and S as a ring member; provided that no more than one of or R ' and R⁸ in -NR'R⁸ is selected from the group consisting of alkoxy, alkylamino, dialkylamino and heterocyclyl;

each n is independently is 0, 1 or 2;

each m is independently 1 , 2, 3 or 4; and

R^1A and R^lB are each independently selected from H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclylaikyi, optionally substituted heterocyclylalkyk optionally substituted aryl, optionally substituted arylaikyi, optionally substituted heteroaryl, or optionally substituted heteroaryialkyl;

or R^1A and R^1B, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5- to 8-membered monocyclic or 5- to 10-membered bicyclic heteroaryl or heterocyclic ring containing up to two additional heteroatoms selected from N, O and S as ring members;

and pharmaceutically acceptable salts, solvates, and/or prodrugs of these compounds.

[00130] in one embodiment of Formula ( I I ) or (IF), the optionally substituted carbocyclyl is an optionally substituted C3-C8 cycloaikyi; the optionally substituted carbocyclylaikyi is an optionally substituted C4-C10 cycloalkylalkyl; and the optionally substituted heteroalkyl is an optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 alkylamino, or optionally substituted C1-C6 dialkylamino.

[00131] In one embodiment of Formula (II) or (IF), -L-M is -NHR⁷, -OR⁷, or -S(0)„R⁷; n is 0, 1, or 2; and R' is optionally substituted C1-C10 alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted arylaikyi, optionally substituted heteroaryialkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted carbocyclylaikyi, or optionally substituted heterocyclylalkyl.

[00132] In one embodiment of Formula (II) or (IF), -L-M is -NR⁷R⁵; and R⁷ and R⁸, taken together with the nitrogen atom to which they are attached, form an optionally substituted hetercyclyi which optionally contains one or more additional heteroatom as ring members.

[00133] In one embodiment of Formula (II) or (IF), -L-M is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocycyl, or optionally substituted heterocyclyl.

[00134] In one embodiment of Formula (II) or (IF), R^lA and R^{l r5} are independently selected from H, optionally substituted C1-C10 alkyl, optionally substituted heterocyclyl, optionally substituted cycloaikyi, optionally substituted cycloalkylalkyl, optionally substituted heterocyclylalkyl, optionally substituted arylalkyl, or an optionally substituted 5-6 membered aryl ring containing up to two heteroatoms as ring members. Preferably the amine group - NR^1AR^1B in compounds of Formulas (II) or (Ila) and IF or Ha' is not -NH₂, -NHMe, or -NMe₂.

[00135] Suitably, R/^A can be selected from H, C1 -C4 alkyl, and C1-C6 acyi, where the alky! and acyl are optionally substituted, In many embodiments, R^1A is H; in other embodiments, it is sometimes Me, or an optionally substituted C1-C4 alkyl. In some embodiments, R^iA is an optionally substituted C1-C6 acyi group, particularly one that can readily be cleaved under mild conditions, such as methoxyacetyl, hydroxyacetyl, or an alpha-arnino acyl group, which can act as pro-drugs for the compounds where R^1A is H.

[00136] Often, R^1A in this amine group -NR^1AR^lB is H, and R.^lB is a substituted or unsubstituted group selected from C2-C8 alkyl, C3-C8 cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, and heterocyclylalkyl. Typically, this aryl is phenyl; heteroaryl refers to a 5-6 membered ring containing up to three heteroatoms selected from N, O and S as ring members; and heterocyclyl refers to a 3-8 membered ring containing at least one heteroatom, and optionally two heteroatoms for 6-8 membered rings, as ring members, where the heteroatoms are selected from , O and S; and the -alkyl- versions of these (arylalkyl, heteroarylalkyl, and heterocyclyl alkyl) typically comprise the specified cyclic group linked via an alkylene linker such as ( ¾)]_-4 to the nitrogen atom of NR^{i A}R^iB. In certain embodiments, R^1B comprises at least one ring having 3-8 ring members,

[00137] Examples of suitable R^iB groups include ethyl, isopropyl, t-butyl, cyclopropyl, cyciobuty], cyclopeniyl, tetrahydrofuranyl, piperidinyl, pyrrol idinyl, cyclopropylmethyl, cyclobutylmethyL phenyl, and the like, each of which can be unsubstituted or substituted with up to three substituents. Some preferred embodiments include cyclopropyl, isopropyl, t-butyl, and cyclo butyl.

[00138] Specific examples of substituted R^1B groups include 2,2,2-trifluoroethyl, 2-methoxy- ethyl, 2-ethoxyethyl, methoxymethyi, 2-aminoethyi, 2-(N-morpholino)ethyl, 3-hydroxypropyl, 3-dimethylaminopropyl, 3-methoxypropyl, 2-hydroxy ethyl, 2-hydroxypropyl, acetyl, benzoyl, phenyl substituted with COOi !, -COO e, -COOEt, ~CONH₂, -CONMe₂, and

where Q represents a functional group such as -OH, -OR, -COOH, -COOR, -NH_?,, -NHR, -N R_.;. -CONH₂, -CONHR, -CONR₂, -SR, ~S(0)R, -S0₂R, -SONR₂, -C(0)R, -NRC(0)R, -NRC(0)OR, -OC(0)OR, -OC(0)NR₂,

wherein each R is independently H or an optionally substituted C1-C4 alkyl group, and two R present on the same functional group can be taken together to form a 5-8 niembered optionally substituted ring, which can contain up to two heteroatoms selected from N, O and S as ring members.

[00139] Where one or more substituents are present on these R, R^1A, or R^lB groups, often the substituents are selected from halo, OR", N(R")₂, S(0)_mR", COOR", CON(R"}₂, CN, phenyl, pyridinyl, pyrrolidinyl, and the like, where each R" is independently selected from H and C 1-C4 alkyl, optionally substituted with one or more groups selected from OI-l, C1 -C4 aikoxy, halo, N¾, C1-C4 alkylamine, and di(Cl-C4)alkyl amine, and piperidine, pyrrolidine, morpholme, or furan; and m is 0-2. Frequently, R^1B comprises at least one ring, such as a heterocylyl or cycloalkyl or aryl ring. A preferred embodiment of R^1B in the amine group -NR^iAR^1B in Formulas (II) and (IV) is cyclopropyl, and a preferred embodiment of R^lA is H.

[00140 J in compounds of Formula (II) and (I V) and (Ila) or ( I la' }, L can be a bond, - CR⁷= R⁸-, -C≡C-, -NR⁷-, -0-, -S(0)_H-, or (CR⁷R^s)_n, or it can be -(CR⁷R⁸)_m-NR⁷-, -(CR⁷R⁸)_ra- 0-, or -(CR⁷R⁸)_m-S(0)_n-. Typically, where L is attached to W at a heteroatom of W, L will be a bond or one of the hydrocarbon linkers, such as (CR^'R )_m. However, embodiments of the invention include compounds wherein -L-W is a group of the formula -NR'-NR'R⁸ as well, Some examples of suitable groups for L include -CH=CH-, -C≡C-, -NH-, NMe, -0-, -S-, - S(0)₂~, and -C¾NH-. Where L is attached to W at a heteroatom of W, L is often CH₂ or (CH₂)₂. [00141] Figures 1 and 2 illustrate the improved selectivity found for compounds of Formula II. Figure 1 depicts a compound of Formula I that is a potent inhibitor of CK2. In assays for inhibition of a panel of 108 kinases, this compound at a comcentration of 1 micromolar is a potent inhibitor of many of the various kinases. By comparison, Figure 2 shows a similar compound of Formula II, ha ving a substituted amine group as an additional substituent on the six-membered ring of the bicyclic core. This compound is more potent as an inhibitor of CK2 than the similar- looking compound in Figure 1; it is less potent as an inhibitor of PIM1; and as the kinase pane! assay shows, it is less potent on many other kinases than the compound of Figure 1 is. Relatively few kinases are inhibited by more than 80% with the amine-substituted compound of Formula II, when compared to the proportion of kinase inhibitors showing similar levels of inhibition by the non-aminated compound of Formula I, This improved selectivity is observed for a wide array of amine substituent groups, as the data in Tables 1 and 2 and additional data throughout the application demonstrate.

[00142] Specific embodiments of the compounds of the invention include compounds of Formula Ha and/or Da':

flla) (Ha')

wherein,

R² is i f CH₃ or CF₃;

Z³ and Z⁴ each independently represent N or CR⁵, or CH;

where each R⁵ is independently selected from halo, -CN, -R, -OR, -S(0)_r.R, -COOR, - CONR², and -NR₂,

wherein each R is independently selected from H and optionally substituted C1-C4 alkyl, or the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5- or 6-memhered heterocyclic ring which contains one or more additional heteroatoffl selected from N, O and S as a ring member;

R is H, CH₃ or CF₃;

X is O, S or NH;

Y is O or S;

R^{5 B} is selected from H, optionally substituted C1-C10 alkyl, optionally substituted heteroaikyi, optionally substituted heterocyclyl, optionally substituted cycloaikyi, optionally substituted cycloalkylalkyl, optionally substituted heterocyclyialkyl, optionally substituted arylalkyl, or an optionally substituted lieteroaryl;

L is a bond, -NR⁷-, -⁽ )-, S((>}_:;-. (CR⁷R⁸)_m, or -(CR⁷R⁸)_m-NR⁷-;

m is 1 , 2, 3, or 4;

n is 0, I , or 2;

W is selected from optionally substituted aryl, optionally substituted heteroaryl, and -NR⁷R^S,

where each R and R ^' is independently selected from H, optionally substituted C1 -C6 alkoxy, optionally substituted C1-C6 alkylamino, optionally substituted C1-C6 diaikyiamino, optionally substituted heterocyclyl optionally substituted CI -C I O alkyl, optionally substituted C3-C8 cycloaikyi, optionally substituted C4-C10 cycloalkylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;

and R' and R , taken together on a single carbon atom or on adjacent connected carbon atoms of (CR^?R⁸)m whether alone or as part of another group, form a 3- to 8-membered ring that contains one or more heteroatoms as ring members;

or R and R⁸, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5- to 10-niernbered heterocyclic or heteroaryl ring system that optionally contains an additional heteroatom selected from N, O and S as a ring member; and

provided that no more than one of or R ' and R⁸ in ~NR⁷R⁸ is selected from the group consisting of alkoxy, alkylamino, diaikyiamino and heterocyclyl.

[00143] In the foregoing compounds of Formula (Ila) or (Ila'), ^z and R⁴ are selected from H, CH₃ and CF₃. In some embodiments R² is H, In some embodiments, R⁴ is H,

[00144] In the foregoing compounds of Formula (Ila) or (Ila'), Y is O or S, In preferred embodiments, Y is O. [00145] In the foregoing compounds of Formula (Ila) or (Ila'), X can be S, O or NH, Frequently, X is NH or 8. In certain embodiments, X is NFL

[00146] In the foregoing compounds of Formula (Ila) or (Ila'), Z^J and 7/ are often selected from and CH. in some embodiments, one of these ring members is N and the other is CFI. In alternative embodiments, both Z³ and Z⁴ are N, In still other embodiments, Z³ and Z^"' are both CH.

[00147] In certain compounds of Formula Ila, Z^J can be N while Z⁴ is CH; or Z³ can be N while Z⁴ is also N. In certain compounds of Formula Ila', Z can be CH while Z⁴ is N;

alternatively, Z^J can be N while Z⁴ is N or CH.

[00148] In the foregoing compounds of Formula (Ila) or (Ila'), R^J, when present, ca be H or optionally substituted alkyl. Often, R³ is H,

[00149] Z can be O or S; in preferred embodiments, Z is O.

[00150] When present, m is frequently 1 or 2,

[00151] In these compounds of Formula Ila and/or Ila', R² and R^" are frequently both H.

[00152] In the foregoing compounds of Formula I l a and/or I l a' , R^{f B} can be optionally substituted CI -CIO alkyl, optionally substituted heterocyciyl, optionally substituted cycioalkyl, optionally substituted cyeloalkylalkyl, optionally substituted heterocyclylalkyl, optionally substituted arylalkyl, or an optionally substituted 5-6 membered aryl ring containing up to two heteroatoms as ring members. In some embodiments, R^llJ is a C3-C6 cycioalkyl or a 3-6 membered heterocyclic group such as piperidine or a C1-C3 alkyl group substituted with one of these rings, and it is optionally substituted. Specific embodiments of R^lB include cyclopropyl, cyclopropvlmethvl, 4-piperidinyI, and substituted 4-piperidinyI, e.g. 4-piperidinyl substituted with an acyl group, such as acetyl, at N-l , Other embodiments include optionally substituted phenyl.

[00153] In the foregoing compounds of Formula (Ila) or (Ila'), -L-M is -NHR⁷, -OR⁷, or - S(0)_nR'; n is 0, 1 , or 2; and R⁷ is optionally substituted C1 -C10 alkyl, optionally substituted heteroalkvl, optionally substituted aryl, optionally substituted lieteroaryl, optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclvl, optionally substituted carbocyclylalkyl, or optionally substituted h eterocy clylalky 1.

[00154] In the foregoing compounds of Formula (Ila) or (Ila' }, -L-M is -NR'R⁸; and R⁷ and R , taken together with the nitrogen atom to which they are attached, form an optionally substituted hetercyclyl which optionally contains one or more additional heteroatom as ring members.

[00155] In the foregoing compounds of Formul a (Ila) or (Ha'), -L- is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carboeycyl, or optionally substituted heterocyciyl.

[00156] in the foregoing compounds of Formula Ila and/or I la', L is typically a bond or NH. When L is NH, W can be an optionally substituted group selected from phenyl, phenylalkyl, heterocyciyl, cycloalkyS and cycloalkylalkyl.

[00157] In the foregoing compounds of Formula Ila and/or Ila', W is frequently an optionally substituted phenyl, arylalkyl, cycloalkyl, heteroaryl, cycloalkylalkyl, or heterocyclic group, Specific examples include optionally substituted phenyl; optionally substituted phenylmethyl; optionally substituted 1-phenylethyl; cyclopropyl methyl; l-cyclopropylethyl; piperidinyl; and morpholinyl. Some preferred substituents for the pheyl groups of W include halo, CN, Me, CF₃, OMe, OCF₃, and heteroaryl groups such as pyrazole or pyrrole or imidazole.

[00158] When L is a bond, W is frequently an optionally substituted aryl, heteroaryl or heterocyciyl group. Surprisingly high flexibility has been demonstrated among the groups that can be represented by W in Formula I I, IF, and (Ila) or (Ila'). Aryl and heteroaryl groups are suitable for W, and can be unsubstituted or substituted. Examples of suitable aromatic groups include phenyl, pyridinyl, pyrimidinyl, thienyl (thiophene ring), furanyl, oxazole, isoxazole, thiazole, isothiazole, oxadiazole, thiadiazole, triazole, and the like, as well as indole,

benzirnidazole, benzofuran, benzopyrazole, imidazole, pyrrole, pyrazole, and the like. Note tha the latter group (indole, benzirnidazole, benzofuran, benzopyrazole, imidazole, pyrrole, pyrazole) contain a 5-membered nitrogen heterocycle, and can be linked to L through either C or N as a result. In some embodiments, W represents one of these aromatic groups that comprises a 5-membered ring, and W is attached via N of the 5-membered ring to L, and L is a bond so that W is effecti vely attached directly to the ring containing Z/^J and 77. Suitable substituents for all of these aryl or heteroaryl groups include those described herein as suitable for such aromatic groups.

[00159] When W is an aromatic group, L is sometimes a bond, NH, or O. A particular embodiment of interest is a compound of Formula II, IF, (Ila) or (Ila'), wherein L is a bond or NH, and W is an optionally substituted phenyl or optionally substituted thienyl ring. In embodiments where L is a bond, it is often desirable for the position of each ring atom of the aryl ring that is adjacent to the attachment point for L to be unsubstituted (i.e., any adjacent carbon(s) would be CH), so the optional substituents on W in such compounds are often, when present, located at positions 3, 4, or 5 of a phenyl ring (assuming position 1 attaches to L), or to positions 4 or 5 of a thienyl ring when L attaches to position 2, and at position 5 of the thienyl group when L attaches at position 3. Examples of these W groups include:

where each A represents the presence of an optional substituent (or more than one where the ring valence permits more) on a carbon not having an explicit H attached,

[00160] Where W is an aromatic group, a wide array of substituents are well tolerated and provide high levels of kinase activity. Suitable substituents include those described herein as suitable for placement on aromatic groups in general. Some of the suitable substituents for these aromatic group W's include halo (especially F or CI), aikyl (e.g., C1-C4 alkyl, such as methyl, ethyl, isopropyl or cyclopropyl); alkoxy (especially C1 -C4 alkyloxy); haioalkyl (e.g., CF₃, -CH₂CF₃); haloalkoxy (e.g. -OCF₃, -OCF₂H, OC¾CF₃, and the like); CN, -OH, alkynyl (e.g., -CCH, CCMe, and the like); heterocyclylmetbyl (e.g., N-piperidinylmethyl,

N-pyrrolidinylmethyl, N-morpholinylmethyl, etc.); hydroxymethyl, aminomethyl,

dimethylaminomethyl, methylaminomethyl; substituted C1-C4 alkoxy such as methoxyethoxy, ethoxymethoxy, trifluoroethoxy, 2-(N-morpholino)ethoxy, 2-( -pyrrolidmyl)ethoxy,

2-(piperidinyl)ethoxy, and the like; acyl groups of the formula -C(Q)-X, where X represents -OR, -NR_2s or ~-R, where each R is independently selected from H or an optionally substituted member selected from C1-C4 alkyl, 3-8 membered cycloalkyl or heterocyclyl, and 5-6 membered aryl or heteroaryl containing up to 3 heteroatoms selected from , O and S as ring members, and where two R on one group (e.g., two R's of-NR?) can be taken together to form an optionally substituted 5-8 membered ring containing up to two heteroatoms selected from N, O and S as ring members; heterocyclic groups such as morpholine, tetrahydrofuran, piperidine, pyrrolidine, 4-Me-N-piperazinyl, N-piperazinyl, 4-acelyl-N-piperazinyl, and the like.

[00161] Commonly, an aromatic group W will have 1-2 substituents, or it will be

unsubstituted; and commonly the substituents, when present, are positioned as described above, so that the ring carbon(s) adjacent to where L is attached are unsubstituted (CH). When L is other than a bond, the substituents on W ca be at any position, and often will be at the positions ortho and/or para to the point of attachment of W to L.

[00162] Alternatively, W ca be a heterocyclic group such as piperidinyi, morpholinyi, pyrrol idinyl, tetrahydrofuranyi, tetrahydropyranyi, thiomorpholinyl, piperazinyl, thiolanyl, and the like, each of which can be unsubstituted or substituted with up to four substituents. Suitable substituents for these groups include those described herein as suitable for heterocyclic groups. Note that even when L is NR or NH, W can be a heterocyclic groups such as 1 -piperidinyi or 4- morpholinyi where L links to a heteroatom (N) of the heterocyclic group as well as at C of the heterocyclic group.

[001 3] Where L is NH, W can also be arylalkyl or cycloalkylalkyl or heterocyclylalkyl, and the alkyl portion of W can be e.g. C1-C4. Where L comprises an alkyl portion, it can be a straight chain (e.g., ethylene, propylene, butylene), or it can be a substituted alkylene chain, resulting in formation of a potentially chiral carbon linker. Where L is a chiral group of this type, e.g. when L is -CH(R)- or -CH₂-CH(R)- where R is not H (e.g., R is Methyl or ethyl, L can be either in an R configuration or an S configuration, where those terms are used in their conventional stereochemical sense, or it can be present as a mixture of isomers, including a racemic mixture. In some embodiments, such a chiral center present in L will be in the S configuration. In other embodiments, it can be in the R configuration,

[00164] Alternatively, W can be a group of the formula or -NR/^'R⁸, -OR⁷, S(0)_nR⁷,

CONR'R⁸, or CR^'R⁸R^y, where each R' and R⁸ and R⁹ is independently selected from H, optionally substituted CI -CIO alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or and R taken together with the N of-NR'R⁸ can form an optionally substituted 5-10 membered heterocyclic or heteroaromatic ring system that optionally contains an additional heteroatom selected from N, O and S as a ring member.

7 8 " 8

[00165] in embodiments where W is -NR R' , L is frequently a bond, and R^' and R taken together with the N of-NR⁷R⁸ can form an optionally substituted 5-10 membered heterocyclic or heteroaromatic ring system that optionally contains an additional heteroatom selected from N, O and S as a ring member. Suitable such rings include e.g., pyrrolidinyl, piperidinyi, piperazinyl, thiomorpholinyl, diazepinyl, and morpholinyi, each of which can be substituted to the extent substitution forms relatively water-stable structures. Suitable substituents include, for example, oxo (=0), C1-C4 alkyl, -OH, -CN, halo (especially F or CI), COOR, CONR₂, SR, -S(0)R, -S0₂R, -NR.₂, hydroxyalkyl, -OR, methoxyalkyl (e.g., methoxymethyS), where each R is independently H or optionally substituted C1-C4 alkyl, and where two R on one group can be taken together to form an optionally substituted 5-8 membered ring containing up to two heteroatoms selected from N, O and S as ring members,

[00166J in some embodiments of the compounds of Formula (ΪΙ) and (IP) and (Ha) or (Ha'), - L-W is a group of the formula -NH-Ar, where Ar represents an op tionally substitu ted aromatic group. Suitable aromatic rings for this group include phenyl, naphthyl, pyridinyl, pyrimidinyi thienyl (thiophene ring), furanyl, indolyl, benzofuranyl, benzothienyl, benzopyrazolyl, benzimidazolyl, benzoxazole, benzothiazole, and the like. Suitable substituents for these aryl or heteroaryl groups include those described herein as suitable for such aromatic groups.

[00167] In some embodiments, W is an optionally substituted cycloaikyi group, typically containing 3-8 ring atoms in a monocyclic structure, or 8- 10 ring atoms in a bicyclic structure. Examples include 1 ,2,3,4-tetrahydronaphth-l-yl, eyclopropyi, cyclobutyl, cyciopentyl, cyclohexyl, decalin, and the like. These groups are optionally substituted as described herein; in some embodiments, the cycloaikyi ring will be substituted with one or more (e.g., up to three) groups selected from halo, hydroxy, oxo ( O h COOR, CONR₂, SR. -S(0)R, -S0₂R, -NR₂, hydroxyalkyl, -OR, methoxyalkyl (e.g., methoxymethyl), C 1-C4 alkyl, where each R is independently H or optionally substituted C1 -C4 alkyl, and where two R on one group can be taken together to form an optionally substituted 5-8 membered ring containing up to two heteroatoms selected from N, O and S as ring members.

[00168] Particular embodiments of the compounds of the inventi on include thiophene- containing compounds of Formul a (II-Th) and (ii-Th'j:

( 11-Th) (II-Tlr)

where R^Tn is selected from H, halo, optionally substituted C 1-C6 alkyl, CN, SfG j , - SO2NR2, COOR, CO R2, and C(0)R, where each R is independently H, halo, CN, or an optionally substituted member selected from the group consisting of C1-C6 alkyl, C1-C6 alkoxy, C1-C6 aiky [amino, di(C 1 - C6)alkylamino, C3-C8 cycloaikyi, C4-C10 cycloalkylalkyl, C5-C8 heterocyclyl, C6-C10 heterocyclylalkyl, ary!, arylalkyl, C5-C6 heteroalkyl, and C6-C10 heteroaikyialkyl;

and two R on the same atom or adjacent connected atoms can form an optionally substituted heterocyciic ring that can contain an additional heteroatom selected from N, 0 and S as a ring member:

and other structural features are as defined for Formula lia above.

[00169] The thienyl (thiophene) ring in Formuias II-Th and II-Th' can be attached to the bicyclic core at either position 2 or position 3 of the thiophene ring, when the position substituted with R ^{, h} is defined as position 5, and the ring sulfur is position 1. In some embodiments, connection is at position 2 of the thienyl group, and in alternative embodiments, connection is at position 3 of the thienyl group,

[00170] In these compounds of Formulas II-Th and II-Th', R² and R⁴ are frequently both H.

[00171] in the foregoing compounds of Formulas II-Th and II-Th', X is preferably NH.

[00172] In die foregoing compounds of Formulas II-Th and II-Th', Y is frequently O.

[00173] In the foregoing compounds of Formulas II-Th and II-Th', Z³ is often N.

[00174] In the foregoing compounds of Formulas II-Th and II-Th', Z⁴ can be CH or N.

[00175] In the foregoing compounds of Formulas II-Th and II-Th', R^1B can be optionally substituted CI -CIO alkyl, optionally substituted heterocyclyl, optionally substituted cycloaikyi, optionally substituted cycloalkylalkyl, optionally substituted heterocyclylalkyl, optionally substituted arylalkyl, or an optionally substituted 5-6 membered aryl ring containing up to two heteroatoms as ring members. In some embodiments, R^llJ is a C3-C6 cycloaikyi or a 3-6 membered heterocyclic group such as piperidine or a C1 -C3 alkyl group substituted with one of these rings, and it is optionally substituted. Specific embodiments of R^1B include cyclopropyl, cyclopropylmethyi, 4-piperidinyl, and substituted 4-piperidinyl, e.g. 4-piperidinyl substituted with an acyl group, such as acetyl, at N-1. Other embodiments include optionally substituted phenyl.

[00176] In these compounds, R^1H can be halo (F, CI, Br), CF₃, CN, C 1-C6 alkyl, C1-C3 alkyl substituted with heterocyclyl or heterocyclylamino, CQOR, or COO^'NR.₂.

[ 00177] In one eniobidment of the present invention, the compounds of Formul a (Ila) or (Ila') have structural Formula (lib) or (lib'):

(lib) (lib')

wherein

and R⁴ are independently H, CH₃ or CF₃;

Z⁴ is N or CH;

-L-M is -NR^8AR⁷, -NHR⁷, -OR⁷, or -8(G)_:1R⁷;

n is 0, 1 , or 2; and

R' is optionally substituted CI -CIO alkyl, optionally substituted lieteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted arylalkyl, optionally substituted iieteroarylalkyl, optionally substituted carbocyclyi, optionally substituted

heterocyclyl, optionally substituted carbocyclylalkyl, or optionally substituted heterocyclyl alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocycyl, or optionally substituted heterocyclyl; or

R' and R^XA, taken together with the nitrogen atom to which they are attached, form an optionally substituted hetercyclyl which optionally contains one or more additional heteroatom as ring members.

[00178] In one emobidment of the present invention, the compounds of Formula (II) have structural Formula (1 Ic)

wherein,

X is O, S, or NR²;

R³ is -(CH₂)-X^C;

X^1" is hydroxyl or a group having stnictura! formula (a), (b), (e), or (d):

L¹ and L² are each independently a covalent bond, -0-, or N R ^":··:

R ^la and R"^a are each independently hydrogen, alkyl, heteroalkyl, heteroaryl, heterocyciyl, alkenyl, alkyny!, arylalkyl, heieroaryialkyl, heterocyclySalkyl, -alkylene-C(Q)-0-R^4a, or -alkylene-G-C(G)-0-R^¾; and

R^3i! and R^4a are each independently hydrogen, alkyl, heteroalkyl, cyclyialkyl,

heterocyciyl, aryl, heteroai l, alkenyl, alkynyl, arylalkyl, heterocyclylalkyl, or heteroarylalkyl;

L^"' is a covalent bond or a!ky!ene;

Y is OR^5a, NR^5aR^6a, or C(0)OR^7a, provided that when Y is C(0)OR^7a, then L³ is not a covalent bond; and

R^5a, R^oa, and R'^a are each independently hydrogen, alkyl, arylalkyl, aryl, heteroalkyl, aiky!heteroaryl, heterocyciyl, or heteroaryl; or alternatively, R^ and R^6a, taken together with the nitrogen atom to which they are attached, form a hetercyclyl ring optionally containing one o rmore additional heteroatom independently selected from N, O, and S.

[00179J In one embodiment of Formula lie), X is NR²; R³ is -(CH₂)-X^C; and X^c is hydroxyl or a group having structural formula (b):

[00180] In one embodiment of Formula (l ie), R" and R⁴ are hydrogen.

[00181] In one embodiment of Formula ( Sic), R^{i B} is an optionally substituted C 1-C10 alkyl, cycloalkyl, or cycloalkylalkyl.

[00182] In one embodiment of Formula (lie), -L-W is -OR ' or X R R^".

[00183] In one embodiment of Formula (lie), R' is optionally substituted aryl or optionally substituted heteroaryl; and R⁸ is H.

L84] In one embodiment of Formula (lie), R^l is optionally substituted phenyl. [00185] In one embodiment of Formula (lie), If is a covalent bond; and Y is QR^yd or NR^5aR^6a.

[00186] The compounds of the invention al so include those enriched in isotopes of the atoms involved in the structures described herein. For example, the compounds as described are intended to include versions wherein one or more H atoms is preferentially enriched in a heavier hydrogen isotope ( deuterium or tritium). In particular, where any of the foregoing compounds contains a methyl group (Me), an enriched methyl group containing deuterium at levels far above natural abundance can be used. For example, -CH₃ could be replaced by -CH₂D or - CHD or -CD₃, where each D represents deuterium present in place of ¾ and indicates that D is present instead of ¹H in at least about 50% of the molecules of a sample of the compound. Of particular interest are compounds comprising -N(R)Me or ~-NMe₂, where Me can be present as CD₃. This variation of the compounds described herein is particularly interesting because the presence of CD₃ in place of CH₃ can have a significant effect on rates of metabolism of an N- methyl group, thus a compound comprising CD₃ can have improved pharmacokinetic properties over a non-enriched compound. Accordingly, the alky! groups described herein are intended to include ones enriched in deuterium, and compounds containing a methyl group on N are specifically considered to include a deuterium-enriched methyl group on N.

[00187] The compounds of the invention often have ionizable groups so as to be capable of preparation as salts. In that case, wherever reference is made to the compound, it is understood in the art that a pharmaceutically acceptable salt may also be used. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases are well-known in the art, such as hydrochloric, sulphuric, hydrobromic, acetic, lactic, citric, or tartaric acids for forming acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, various amines, and the like for forming basic salts. Methods for preparation of the appropriate salts are well-established in the art. In some cases, the compounds may contain both an acidic and a basic functional group, in which case they may have two ionized groups and yet have no net charge.

Utilities of the Compounds:

[00188] In another aspect, the invention provides a pharmaceutical composition comprising any of the above-described compounds, admixed with a pharmaceutically acceptable excipient. [00189] In another aspect, the invention provides a method to treat cancer, a vascular disorder, inflammation, infection, pain, or an immunological disorder comprising administering to a subject in need of such treatment, an effective amount of any of the above-described compounds.

[00190] The compounds of the invention are useful as medicaments, and are useful for the manufacture of medicaments, including medicaments to treat conditions disclosed herein, such as cancers, inflammatory conditions, infections, pain, and immunological disorders.

[00191] The terms "treat" and "treating" as used herein refer to ameliorating, alleviating, lessening, and removing symptoms of a disease or condition, A candidate molecule or compound described herein may be in a therapeutically effective amount in a formulation or medicament, which is an amount that can lead to a biological effect, such as apoptosis of certain cells (e.g., cancer cells), reduction of proliferation of certain cells, or lead to ameliorating, alleviating, lessening, or removing symptoms of a disease or condition, for example. The terms also can refer to reducing or stopping a cell proliferation rate (e.g., slowing or halting tumor growth) or reducing the number of proliferating cancer cells (e.g., removing part or all of a tumor).

[00192] These terms also are applicable to reducing a titre of a microorganism in a system (i.e., cell, tissue, or subject) infected with a microorganism, reducing the rate of microbial propagation, reducing the number of symptoms or an effect of a symptom associated with the microbial infection, and/or removing detectable amounts of the microbe from the system.

Exampl es of microorganisms include but are not limited to vims, bacterium and fungus.

[ 00193] The compounds of the invention have activities to modulate protein kinases, in particular C 2 activity and/or Pirn activity. In some embodiments, the compounds of the invention specifically inhibit the activity of C 2, but not Pim, e.g., more than 100, 90, 80, 70, 60. 50, 40, 30, 20, or 10 fold difference between CK2 inhibition vs. Pim inhibition. In some embodiments, the compounds of the invention specifical ly inhibit the acitivitv of Pim, but not Ck2, e.g., more than 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 fold difference between Pim inhibition vs. CK2 inhibition. In some embodiments, the compounds of the invention inhibit the activity of CK2 as well as Pim.

[00194] The compounds of the invention can be used to modulate the activity of CK2 and/or Pim, e.g., inhibit the activity of CK2 and/or Pim in a cell, e.g., in vivo or in vitro. In some embodiments, compounds of the invention can be used to modulate the acti vity of C 2, e.g., inhibit the activity of CK2 without substantially interferring or changing the activity of Pim. In some embodiments, compounds of the invention can be used to modulate the activity of Pirn, e.g., inhibit the activity of Pirn without substantially interferring or changing the activity of CK2. In some embodiments, compounds of the invention can be used to modulate the activity of CK2 and Pirn, e.g., inhibit the activity of C 2 and Pirn.

[00195] The compounds of the invention are thus useful to treat infections by certain pathogens, including protozoans and viruses. The invention thus provides methods for treating protozoal disorders such as protozoan parasitosis, including infection by parasitic protozoa responsible for neurological disorders such as schizophrenia, paranoia, and encephalitis in immunocompromised patients, as well as Chagas' disease, it also provides methods to treat various viral diseases, including huma immunodeficiency virus type 1 (HIV-1), human papilloma viruses (HPVs), herpes simplex virus (HSV), Epstein-Barr virus (EBV), human cytomegalovirus, hepatitis C and B viruses, influenza virus, Borna disease virus, adenovirus, coxsackieviras, coronavims and varicella zoster vims. The methods for treating these disorders comprise administering to a subject in need thereof an effective amount of a compound of Formula II or Formula IF.

[00196] As used herein, the term "apoptosis" refers to an intrinsic ceil self-destruction or suicide program. In response to a triggering stimulus, cells undergo a cascade of events including cell shrinkage, blebbing of cell membranes and chromatic condensation and fragmentation. These events culminate in cell conversion to clusters of membrane -bound particles (apoptotic bodies), which are thereafter engulfed by macrophages.

[00197] The invention in part provides pharmaceutical compositions comprising at least one compound within the scope of the invention as described herein, and methods of using compounds described herein.

[001 8] in addition, the invention in part provides methods for identifying a candidate molecule that interacts with a CK2, which comprises contacting a composition containing a CK2 protein and a molecule described herein with a candidate molecule and determining whether the amount of the molecule described herein that interacts with the protein is modula ted, whereby a candidate molecule that modul ates the amount of the molecule described herein that interacts with the protein is identified as a candidate molecule that interacts with the protein.

[00199] Also provided by the invention are methods for modulating certain protein kinase activities. Protein kinases catalyze the transfer of a gamma phosphate from adenosine triphosphate to a serine or threonine amino acid (serine/threonine protein kinase), tyrosine amino acid (tyrosine protein kinase), tyrosine, serine or threonine (dual specificity protein kinase) or histidine amino acid (histidine protein kinase) in a peptide or protein substrate. Thus, included herein are methods which comprise contacting a system comprising a protein kinase protein with a compound described herein in an amount effective for modulating (e.g. , inhibiting) the activity of the protein kinase. In some embodiments, the activity of the protein kinase is the catalytic activity of the protein (e.g. , catalyzing the transfer of a gamma phosphate from adenosine triphosphate to a peptide or protein substrate), in certain embodiments, provided are methods for identifying a candidate molecule that interacts with a protein kinase, which comprise:

contacting a composition containing a protein kinase and a compound described herein with a candidate molecule under conditions in which the compound and the protein kinase interact, and determining whether the amount of the compound that interacts with the protein kinase is modulated relative to a control interaction between the compound and the protem kinase without the candidate mol ecule, whereby a candidate mol ecule that mod ulates the amount of the compound interacting with the protein kinase relative to the control interaction is identified as a candidate molecule that interacts with the protein kinase. Systems in such embodiments can be a cell-free system or a system comprising cells (e.g. , in vitro). The protein kinase, the compound or the molecule in some embodiments is in association with a solid phase. In certain embodiments, the interaction between the compound and the protein kinase is detected via a detectable label, where in some embodiments the protein kinase comprises a detectable label and in certain embodiments the compound comprises a detectable label. The interaction between the compound and the protein kinase sometimes is detected without a detectable label.

[00200] Provided also are compositions of matter comprising a protein kinase and a compound described herein. In some embodiments, the protein kinase in the composition is a serine-threonine protein kinase. In some embodiments, the protein kinase in the composition is, or contains a subunit (e.g., catalytic subunit, SH2 domain, SH3 domain) of, C 2. In certain embodiments the composition is cell free and sometimes the protein kinase is a recombinant protein.

[00201] The protein kinase can be from any source, such as ceils from a mammal, ape or human, for example. Examples of serine-threonine protein kinases that ca be inhibited, or may potentially be inhibited, by compounds disclosed herein include without limitation human versions of CK2, or CK2a2. A serine-threonine protein kinase sometimes is a member of a subfamily containing one or more of the following amino acids at positions corresponding to those listed in human CK2: leucine at position 45, methionine at position 163 and isoleucine at position 174. Nucleotide and amino acid sequences for protein kinases and reagents are publicly available (e.g. , World Wide Web URLs www.ncbi.nlm.nih.gov/sites/entrez/ and www.Invitrogen.com, each last visited December 2, 2009),

[00202] The invention also in part, provides methods for treating a condition related to aberrant cell proliferation. For example, provided are methods of treating a cell proliferative condition in a subject, which comprises administering a compound described herein to a subject in need thereof in an amount effective to treat the ceil proliferati ve condition. The subject may be a research animal (e.g., rodent, dog, cat, monkey), optionally containing a tumor such as a xenograft tumor (e.g., human tumor), for example, or may be a human. A cell proliferative condition sometimes is a tumor, e.g., solid or circulating tumor or non-tumor cancer, including but not limited to, cancers of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood and heart (e.g., leukemia, lymphoma, carcinoma).

[00203] Compounds and compositions of the invention may be used alone or in combination with anticancer or other agents, such as a palliative agents, that are typically administered to a patient being treated for cancer, as further described herein.

[00204] Also provided are methods for treating a condition related to inflammation or pain. For example, methods are provided for treating pain in a subject, which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the pain. Provided also are methods of treating inflammation in a subject, which comprise administering a compound described herein to a subject in need thereof in an amount effective to treat the inflammation. The subject may be a research animal (e.g., rodent, dog, cat, monkey), for example, or may be a human. Conditions associated with inflammation and pain include without limitation acid reflux, heartburn, acne, allergies and allergen sensitivities, Alzheimer's disease, asthma, atherosclerosis, bronchitis, carditis, celiac disease, chronic pain, Crohn's disease, cirrhosis, colitis, dementia, dermatitis, diabetes, dry eyes, edema, emphysema, eczema, fibromyalgia, gastroenteritis, gingivitis, heart disease, hepatitis, high blood pressure, insulin resistance, interstitial cystitis, joint pain/arthritis/rheumatoid arthritis, metabolic syndrome (syndrome X), myositis, nephritis, obesity, osteopenia, glomerulonephritis (GN), juvenile cystic kidney disease, and type I nephronophthisis (NPHP), osteoporosis, Parkinson's disease, Guam- Parkinson dementia, supranuclear palsy, Kuf s disease, and Pick's disease, as well as memory impairment, brain ischemia, and schizophrenia, periodontal disease, polyarteritis, polychondritis, psoriasis, scleroderma, sinusitis, Sjogren's syndrome, spastic colon, systemic candidiasis, tendonitis, urinary track infections, vaginitis, inflammatory cancer (e.g., inflammatory breast cancer) and the like,

[00205] Methods for detemnning and monitoring effects of compounds herein on pain or inflammation are known. For example, formalin-stimulated pain behaviors in research animals can be monitored after administration of a compound described herein to assess treatment of pain (e.g., Li et al., Pain 115(1-2): 182-90 (2005)). Also, modulation of pro-inflammatory molecules (e.g., IL-8, GRQ-aipha, MCP-1, TNF alpha and iNQS) can be monitored after administration of a compound described herein to assess treatment of inflammation (e.g., Parhar et al, Int J Colorectal Dis. 22(6): 601-9 (2006)), for example. Thus, also provided are methods for determining whether a compound herein reduces inflammation or pain, which comprise contacting a system with a compound described herein in an amount effective for modulating (e.g., inhibiting) the activity of a pain signal or inflammation signal.

[00206] Provided also are methods for identifying a compound that reduces inflammation or pain, which comprise: contacting a system with a compound of Formula II or Formula IF; and detecting a pain signal or inflammation signal, whereby a compound that modulates the pain signal relative to a control molecule is identified as a compound that reduces inflammation of pain. Non-limiting examples of pain signals are formalin-stimulated pain behaviors and examples of inflammation signals include without limitation a level of a pro-inflammatory molecule. The invention thus in part pertains to methods for modulating angiogenesis in a subject, and methods for treating a condition associated with aberrant angiogenesis in a subject, proliferative diabetic retinopathy.

[ 00207] CK2 has also been shown to play a role in the pathogenesis of atherosclerosis, and may prevent atherogenesis by maintaining laminar shear stress flow. CK2 plays a role in vascularization, and has been shown to mediate the hypoxia-induced activation of histone deacetylases (HDACs). CK2. is also involved in diseases relating to skeletal muscle and bone tissue, including, e.g., cardiomyocyte hypertrophy, heart failure, impaired insulin signaling and insulin resistance, hypophosphatemia and inadequate bone matrix mineralization.

[00208] Thus in one aspect, the invention provides methods to treat each of these conditions, comprising administering to a subject in need of such treatment an effect amount of a C 2 inhibitor, such as a compound of Formula II or Formula IF as described herein.

[00209] The invention also in part pertains to methods for modulating an immune response in a subject, and methods for treating a condition associated with an aberrant immune response in a subject. Thus, provided are methods for determining whether a compound herein modulates an immune response, which comprise contacting a system with a compound described herein in an amount effective for modulating (e.g., inhibiting) an immune response or a signal associated with an immune response. Signals associated with immunomodulatory activity include, e.g. , stimulation oi^' 1 -cell proliferation, suppression or induction of cytokines, including, e.g., interleukins, interferon-γ and TNF, Methods of assessing immunomodulatory activity are known in the art,

[00210J Also provided are methods for treating a condition associated with an aberrant immune response in a subject, which comprise administering a compound described herein to a subject in need thereof in an amount effecti ve to treat the condition. Conditions characterized by an aberrant immune response include without limitation, organ transplant rejection, asthma, autoimmune disorders, including rheumatoid arthritis, multiple sclerosis, myasthenia gravis, systemic lupus erythematosus, scleroderma, polymyositis, mixed connective tissue disease (MCTD), Crohn's disease, and ulcerative colitis. In certain embodiments, an immune response may be modulated by administering a compound herein in combination with a molecule that modulates (e.g., inhibits) the biological activity of an mTOR pathway member or member of a related pathway (e.g., mTOR, PI3 kinase, AKT). In certain embodiments the molecule that modulates the biological activity of an mTOR pathway member or member of a related pathway is rapamycin. In certain embodiments, provided herein is a composition comprising a compound described herein in combination with a molecule that modulates the biological activity of an mTOR pathway member or member of a related pathway, such as rapamycin, for example.

Compositions and Routes of Administration

[00211] In another aspect, the invention provides pharmaceutical compositions (i.e., formulations). The pharmaceutical compositions can comprise a compound of any of Formulae (I), (II), (IF), (Ila), (Ha'), (lib), (lib'), (II-Th), and (II-Th'), as described herein which is admixed with at least one pharmaceutically acceptable excipient or carrier. Frequently, the composition comprises at least two pharmaceutically acceptable excipients or carriers,

[00212 J While the compositions and methods of the present invention wi ll typical ly be used in therapy for human patients, they may also be used in veterinary medicine to treat similar or identical diseases. The compositions may, for example, be used to treat mammals, including, but not limited to, primates and domesticated mammals, The compositions may, for example be used to treat herbivores. The compositions of the present invention include geometric and optical isomers of one or more of the drugs, wherein each drug is a racemic mixture of isomers or one or more purified isomers,

[00213] Pharmaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose. Determination of the effective amounts is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.

[00214] The compounds of the present invention may exist as pharmaceutically acceptable salts. The present invention includes such salts. The term "pharmaceutically acceptable salts" is meant to include salts of active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituent moieties found on the compounds described herein. When compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent, included are base addition salts such as sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrohromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids, for example, acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthaiic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1 -19). Certain specific compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts,

[00215] Examples of applicable salt forms include hydrochlorides, hydrobromides, sulfates, methanesulfoTiates, nitrates, maleates, acetates, citrates, fumarates, tartrates (eg (+)-tartrates, (-)- tartrates or mixtures thereof, including racemic mixtures), succinates, benzoates and salts with amino acids such as glutamic acid. These salts may be prepared by methods known to those skilled in art. [00216] The neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.

[00217] The pharmaceutically acceptable esters in the present invention refer to non-toxic esters, preferably the alkyl esters such as methyl, ethyl, propyl, isopropyS, butyl, isobutyl or pentyl esters, of which the methyl ester is preferred. However, other esters such as phenyl-Cj-s alkyl may be employed if desired. Ester derivatives of certain compounds may act as prodrugs which, when absorbed into the bloodstream of a warm-blooded animal, may cleave in such a manner as to release the drug form and permit the drug to afford improved therapeutic efficacy.

[00218] Certain compounds of the present invention can exist in irasolvated forms as well as solvated forms, including hydrated forms. In general, the soivated forms are equivalent to unsolvated forms and are encompassed within the scope of the present invention. Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms are ecjuivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

[00219] When used as a therapeutic the compounds described herein often are administered with a physiologically acceptable carrier. A physiologically acceptable carrier is a formulation to which the compound can be added to dissolve it or otherwise facilitate its administration. Examples of physiologically acceptable carriers include, but are not limited to, water, saline, physiologically buffered saline.

[ 00220] Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of a hy drogen by a deuterium or tritium, or the replacement of a carbon by ^l3C- or ^C-enriched carbon are within the scope of this invention. The compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of atoms that constitute such compounds. For example, the compounds may be radiolabeled with radioactive isotopes, such as for example tritium (3H), iodine- 125 ( ⁵I) or carbon- 14 (^{¾ 4}C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.

[00221 ] In addition to salt forms, the present invention provides compounds that are in a prodrug form. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present invention, Additionally, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an e vivo environment, For example, prodrugs can be slowly con verted to the compounds of the present invention when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[00222] A compound of the present in vention can be formulated as a pharmaceutical composition. Such a pharmaceutical composition can then be administered orally, pareiiteraily, by inhalation spray, rectally, or topically in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles as desired. The amount of active ingredient that ca be combined with the carrier materials to produce a single dosage form varies depending upon the mammalian host treated and the particular mode of

administration, Topical administration can also involve the use of transdermal administration such, as transdermal patches or iontophoresis devices. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrastemal injection, or infusion techniques. Formulation of drugs is discussed in, for example, Hoover, John E., REMI NGTON'S PHARMACEUTICAL SCIENCES, Mack Publishing Co,, Easton, Pa,; 1975. Other examples of drug formulations can be found in Liberman, H. A. and Lachman, L., Eds., PHARMACEUTICAL DOSAGE FORMS, Marcel Decker, New York, N.Y., 1980.

[00223] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1 ,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water. Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or digiycerides. in addition, fatty acids such as oleic acid find use in the preparation of injectables. Dimethyl acetamide, surfactants including ionic and non-ionic detergents, polyethylene glycols can be used. Mixtures of solvents and wetting agents such as those discussed above are also useful.

[00224] Suppositories for rectal administration of the drug ca be prepared by mixing the drug with a suitable nonirritating excipient such as cocoa butter, synthetic mono- di- or triglycerides, fatty acids and polyethylene glycols that are sold at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum and release the drug. [00225] Solid dosage forms for oral administration can include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the compounds of this invention are ordinarily combined with one or more adjuvants appropriate to the indicated route of

administration. If administered per os, a compound of the invention can be admixed with lactose, sucrose, starch powder, cellulose esters of alkaiioic acids, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, gelatin, acacia gum, sodium alginate, polyvinylpyrrolidone, and/or polyvinyl alcohol, and then tableted or encapsulated for convenient administration. Such capsules or tablets can contain a controlled-release formulation as can be provided in a dispersion of active compound in hydroxypropylmethyl cellulose. In the case of capsules, tablets, and pills, the dosage forms can also comprise buffering agents such as sodium citrate, magnesium or calcium carbonate or bicarbonate. Tablets and pills can additionally be prepared with enteric coatings,

[00226] For therapeutic purposes, formulations for parenteral administration can be in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. These solutions and suspensions can be prepared from sterile powders or granules having one or more of the carriers or diluents mentioned for use in the formulations for oral adminis tration. A compound of the invention can be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, and/or various buffers. Other adjuvants and modes of administration are well and widely known in the pharmaceutical art.

[00227] Liquid dosage forms for oral administration can include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions can also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents.

[00228] The dosage regimen utilizing the compounds of the present invention in combination with an anticancer agent is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt or ester thereof employed. A consideration of these factors is well within the purview of the ordinarily skilled clinician for the purpose of determining the therapeutical ly effective dosage amounts to be given to a person in need of the instant combination therapy. [00229] [I believe these paragraphs are repeats of paragraph 0184-0187] In certain

embodiments of the present invention, the compound is a compound of Formul a (I)a, and in certain embodiments it is a compound of Formula (!)b.

[00230 J Any suitable formulation of a compound described above can be prepared for administration by methods known in the art. Selection of useful excipieiits or carriers can be achieved without undue experimentation, based on the desired route of administration and the physical properties of the compound to be administered.

[00231 J Any suitable route of administration may be used, as determined by a treating physician, including, but not limited to, oral, parenteral, intravenous, intramuscular, transdennai topical and subcutaneous routes. Depending on the subject to be treated, the mode of administration, and the type of treatment desired— e.g., prevention, prophylaxis, therapy; the compounds are formulated in ways consonant with these parameters. Preparation of suitable formulations for each route of administration are known in the art. A summary of such formulation methods and techniques is found in Remington's Pharmaceutical Sciences, latest edition, Mack Publishing Co., Easton, PA. The formulation of each substance or of the combination of two substances will frequently include a diluent as well as, in some cases, adjuvants, buffers, preservatives and the like. The substances to be administered can be administered also in liposomal compositions or as microemulsions.

[00232] For injection, formulations can be prepared in conventional forms as liquid solutions or suspensions or as solid forms suitable for solution or suspension in liquid prior to injection or as emulsions. Suitable excipienis include, for example, water, saline, dextrose, glycerol and the like. Such compositions may also contain amounts of nontoxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, such as, for example, sodium acetate, sorbitan monoiaurate, and so forth.

[00233] Various sustained release systems for drugs have also been devised, and can be applied to compounds of the invention. See, for example, U.S. patent No. 5,624,677, the methods of which are incorporated herein by reference.

[00234] Systemic administration may also include relatively noninvasive methods such as the use of suppositories, transdermal patches, transniucosal delivery and intranasal administration. Oral administration is also suitable for compounds of the invention. Suitable forms include syrups, capsules, tablets, as is understood in the art.

[00235] For administration to animal or human subjects, the appropriate dosage of a compound described above often is 0.01-15 mg/kg, and sometimes 0.1-10 mg/kg. in some embodiments, a suitabl e dosage of the compound of the invention for an adult patient will be between 1 and 1000 mg per dose, frequently between 10 and 300 mg, and the dosage may be administered 1 -4 times per day, Dosage levels are dependent on the nature of the condition, drug efficacy, the condition of the patient, the judgment of the practitioner, and the frequency and mode of administra tion; optimization of such parameters is within the ordinary level of skill in the art.

Therapeutic Combinations:

[00236] Compounds of the invention may be used alone or in combination with another therapeutic agent. The invention provides methods to treat conditions such as cancer, inflammation and immune disorders by administering to a subject in need of such treatment a therapeutically effective amount of a therapeutic agent useful for treating said disorder and administering to the same subject a therapeutically effective amount of a modulator of the present invention, i.e., a compound of the invention. The therapeutic agent and the modulator may be "co-administered", i.e, administered together, either as separate pharmaceutical compositions or admixed in a single pharmaceutical composition. By "administered together", the therapeutic agent and the modulator may also be administered separately, including at different times and with different frequencies. The modulator may be administered by any- known route, such as orally, intravenously, intramuscularly, nasally, and the like; and the therapeutic agent may also be administered by any conventional route. In many embodiments, at least one and optionally both of the modulator and the therapeutic agent may be administered orally. Preferably, the modulator is an inhibitor, and it may inhibit either one of CK2 and Pim, or both of them to provide the treatment effects described herein.

[00237] In certain embodiments, a "modulator" as described above may be used in combination with a therapeutic agent that can act by binding to regions of DNA that can form certain quadruplex structures. In such embodiments, the therapeutic agents have anticancer activity on their own, but their activity is enhanced when they are used in combination with a modulator. This synergistic effect allows the therapeutic agent to be administered in a lower dosage while achieving equivalent or higher levels of at least one desired effect.

[00238] A modulator may be separately active for treating a cancer. For combination therapies described above, when used in combination with a therapeutic agent, the dosage of a modulator will frequently be two-fold to ten-fold lower than the dosage required when the modulator is used alone to treat the same condition or subject. Determination of a suitable amount of the modulator for use in combination with a therapeutic agent is readily determined by methods known in the art.

[00239] Compounds and compositions of the invention may be used in combination with anticancer or other agents, such as palliative agents, that are typically administered to a patient being treated for cancer. Such "anticancer agents" include, e.g., classic chemotherapeutic agents, as well as molecular targeted therapeutic agents, biologic therapy agents, and

radiotherapeutic agents .

[00240 J When a compound or composition of the in venti on is used in combinati on with an anticancer agent to another agent, the present invention provides, for example, simultaneous, staggered, or alternating treatment. Thus, the compound of the invention may be administered at the same time as an anticancer agent, in the same pharmaceutical composition; the compound of the invention may be administered at the same time as the anticancer agent, in separate pharmaceutical compositions; the compound of the invention may be administered before the anticancer agent, or the anticancer agent may be administered before the compound of the invention, for example, with a time difference of seconds, minutes, hours, days, or weeks.

[00241] In examples of a staggered treatment, a course of therapy with the compound of the in vention may be administered, followed by a course of therapy with the anticancer agent, or the reverse order of treatment may be used, and more than one series of treatments with each component may also be used, in certain examples of the present invention, one component, for example, the compound of the invention or the anticancer agent, is administered to a mammal while the other component, or its derivative products, remains in the bloodstream of the mammal. For example, the present compound may be administered while the anticancer agent or its derivative products remains in the bloodstream, or the anticancer agent may be

administered while the present compound or its derivatives remains in the bloodstream. In other examples, the second component is administered after ail, or most of the first component, or its derivatives, have left the bloodstream of the mammal.

[00242] The compound of the invention and the anticancer agent may be administered in the same dosage form, e.g., both administered as intravenous solutions, or they may be administered in different dosage forms, e.g., one compound may be administered topically and the other orally. A person of ordinary skill in the art would be able to discern which combinations of agents would be useful based on the particular characteristics of the drugs and the cancer involved. [00243] Anticancer agents useful in combination with the compounds of the present invention may include agents sel ected from any of the classes known to those of ordinary skill in the art, including, but not limited to, antirnicrotubule agents such as diterpenoids and vinca alkaloids; platinum coordination complexes; alkylating agents such as nitrogen mustards,

oxazaphosphorines, alkylsuifonates, nitrosoureas, and triazenes; antibiotic agents such as anthracyclins, actinomycins and bleomycins; topoisomerase II inhibitors such as

epipodophyilotoxins; antimetabolites such as purine and pyrimidine analogues and anti-folate compounds; topoisomerase I inhibitors such as camptothecins; hormones and hormonal analogues; signal transduction pathway inhibitors; nonreceptor tyrosine kinase angiogenesis inhibitors; immunotherapeutic agents; pro-apoptotic agents; and cell cycle signaling inhibitors; and other agents described below,

[00244] Anti-microtubuie or anti-mitotic agents are phase specific agents that are typically active against the microtubules of tumor cells during M or the mitosis phase of the cell cycle. Examples of anti-microtubuie agents include, bu t are not limited to, diterpenoids and vinca alkaloids.

[00245] Plant alkaloid and terpenoid derived agents include mitotic inhibitors such as the vinca alkaloids vinblastine, vincristine, vindesine, and vinoreibine; and microtubule polymer stabilizers such as the taxanes, including, but not limited to paclitaxel, docetaxel, larotaxel, ortataxel, and tesetaxel.

[00246] Diterpenoids, which are derived from natural sources, are phase specific anti - cancer agents that are believed to operate at the G2/M phases of the cell cycle. It is believed that the diterpenoids stabilize the p-tubulin subunit of the microtubules, by binding with this protein. Disassembly of the protein appears then to be inhibited with mitosis being arrested and cell death following.

[00247] Examples of diterpenoids include, but are not limited to, taxanes such as paclitaxel, docetaxel, larotaxel, ortataxel, and tesetaxel. Paclitaxel is a natural diterpene product isolated from the Pacific yew tree Taxus brevifolia and is commercially available as an injectable solution TAXOL®. Docetaxel is a semisynthetic derivative of paclitaxel q. ., prepared using a natural precursor, 10-deacetyl-baccatin III, extracted from the needle of the European Yew tree. Docetaxel is commercially available as an injectable solution as TAXOTERE®,

[00248] Vinca alkaloids are phase specific anti-neoplastic agents derived from the periwinkle plant. Vinca alkaloids that are believed to act at the M phase (mitosis) of the cell cycle by binding specifically to tubulin. Consequently, the bound tubulin molecule is unable to polymerize into microtubules. Mitosis is believed to be arrested in metaphase with cell death following, Examples of vinca alkaloids include, but are not limited to, vinblastine, vincristine, vindesine, and vinorelbine, Vinblastine, vincaleukoblastine sulfate, is commercially available as VELBAN® as an injectable solution. Vincristine, vincaleukoblastine 22-oxo-sulfate, is commercially available as ONCOVIN® as an injectable solution. Vinorelbine, is commercially available as an injectable solution of vinorelbine tartrate (NA VELBINE®), and is a

semisynthetic vinca alkaloid derivative.

[00249 J Platinum coordination complexes are non-phase specific anti-cancer agents, which are interactive with DNA. The platinum complexes are believed to enter tumor cells, undergo, aquation and form intra- and interstrand crosslinks with DNA causing adverse biological effects to the tumor. Platinum-based coordination complexes include, but are not limited to cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, and (SP-4~3)-(cis)- ammmedichloro-[2- methylpyridine] platinum(II). Cisplatin, cis-diamminedichloroplatinum, is commercially available as PLATINOL® as an injectable solution. Carboplatin, platinum, diammine [1, 1- eyclobutane-dicarboxylate(2~)~0,0'], is commercially available as PARAPLAT1N® as an injectable solution.

[00250 J Alkylating agents are generally non-phase specific agents and typically are strong electrophiles. Typically, alkylating agents form covalent linkages, by alkylation, to DNA through nucleophilic moieties of the DNA molecule such as phosphate, amino, sulfhydryl, hydroxy!, carboxyl, and imidazole groups. Such alkylation disrupts nucleic acid function leading to cell death. Examples of alkylating agents include, but are not limited to, alkyl sulfonates such as busulfan; ethyleneimine and methylmelamine derivatives such as altretamme and thiotepa; nitrogen mustards such as chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechiorethamine, melphalan, and uramustine; nitrosoureas such as carmustine, lomustine, and streptozocin; triazenes and imidazotetrazines such as dacarbazine, procarbazine, temozolamide, and temozolomide. Cyclophosphamide, 2-[bis(2-chloroethyl)-amino]tetrahydro- 2H- 1 ,3 ,2-oxazaphosphorine 2-oxide monohydrate, is commercially available as an injectable solution or tablets as CYTOXAN®, Melphalan, 4-[bis(2-chloroethyl)aminoj-L-phenylalanine, is commercially available as an injectable solution or tablets as ALKERAN®. Chlorambucil, 4- [bis(2-chloroemyl)ammo]-benzenebutanoic acid, is commercially available as LEUKERAN® tablets. Busulfan, 1 ,4-butanediol dimethanesulfonate, is commercially available as

YLERAN® TABLETS. Carmustine, l ,3-[bis(2-chloroethyl)-l -nitrosourea, is commercially available as single vials of lyophilized material as BiCNU®, 5-(3,3-dimethyl- 1 -triazeno)- imidazole ~4~earboxamide, is commercially available as single vials of material as DTIC- Dome®. Furthermore, alkylating agents include (a) alkylating-like platinum-based

chemotherapeutic agents such as cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, and (SP-4-3)-(cis)-amminedichloro-[2-methylpyridine] platinum(fl); (b) alky! sulfonates such as busulfan; (c) ethyleneimine and methylmelamine derivatives such as altretamine and tbiotepa; (d) nitrogen mustards such as chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechiorethamine, trofosamide, prednimustine, melphalan. and uramustine; (e) nitrosoureas such as carmustine, lomustine, fotemustine, nimustine, ranimustine and streptozocin; (f) t iazenes and imidazotetrazines such as dacarbazine, procarbazine, temozolamide, and temozolomide.

[0Θ251] Anti-tumor antibiotics are non-phase specific agents which are believed to bind or intercalate with DNA. This may result in stable DNA complexes or strand breakage, which disrupts ordinary function of the nucleic acids, leading to cell death. Examples of anti-tumor antibiotic agents include, but are not limited to, anthracyclines such as daunorubicin (including liposomal daunorubicin), doxorubicin (including liposomal doxorubicin), epirubicin, idarubicin, and valrubicin; streptomyces-related agents such as bleomycin, actinomycin, mithramycin, mitomycin, porfiromycin: and mitoxantrone, Dactinomycin, also know as Actinomycin D, is commercially available in injectable form as COSMEGEN®. Daunorubicin, (8S-cis-)-8-acetyl- 10-[(3-amino-2,3,6 rideoxy-a-L-lyxohexopyranosyl)oxy]-7,8,9,10-tetrahydro-6,8, 11- trihydroxy- 1 -methoxy-5, 12-naphthacenedione hydrochloride, is commercially available as a liposomal injectable form as DAUNOXOME® or as an injectable as CERUBIDINE®.

Doxorubicin, (8S, 10S)-iO-[(3~amino-2,3,6-trideoxy-a-L-lyxohexopyranosyl)oxy]~8-glycok)yi, 7,8,9,1 0-tetrahydro-6,8, 1 1-trihydroxy-l -methoxy-5, 12-naphthacenedione hydrochloride, is commercially available in an injectable form as RUB EX® or ADRIAMYCIN RDF®.

Bleomycin, a mixture of cytotoxic glycopeptide antibiotics isolated from a strain of

Streptomyces verticil/us, is commercially available as BLENOXANE®.

[00252 J Topoisomerase inhibitors include topoisomerase I inhibitors such as camptothecin, topotecan, irinotecan, rubitecan, and belotecan; and topoisomerase II inhibitors such as etoposide, teniposide, and amsacrine,

[00253] Topoisomerase II inhibitors include, but are not limited to, epipodophyllotoxins, which are phase specific anti-neoplastic agents derived from the mandrake plant.

Epipodophyllotoxins typically affect cells in the S and G2 phases of the cell cycle by forming a ternary complex with topoisomerase I I and DN A causing DNA strand breaks. The strand breaks accumulate and cell death follows. Examples of epipodophyllotoxins include, but are not limited to, etoposide, tenyposide, and amsacrine. Etoposide, 4'-demethyl-epipodophyllotoxin 9[4,6-0-(R)-ethylidene-p-D- glucopyranoside], is commercially available as an injectable solution or capsules as VePESID® and is commonly known as VP-16. Tenyposide, 4'~demethyl- epipodophyllotoxin 9[4,6-0-(R )-thenylidene- -D-glucopyranoside], is commercially available as an injectable solution as VUMON© and is commonly known as VM-26,

[00254] Topoisomerase I inhibitors including, camptothecin and camptothecin derivatives. Examples of topoisomerase I inhibitors include, but are not limited to camptothecin, topotecan, irinotecan, rubitecan, belotecan and the various optical forms (i.e., (R), (S) or (R,S)) of 7-(4- methylpiperazino-methy lene)- 10, 1 l-ethylenedioxy-camptothecin, as described in U.S. Patent Nos. 6,063,923; 5,342,947; 5,559,235; 5,491 ,237 and pending U.S. patent Application No. 08/977,217 filed November 24, 1997. Irinotecan HCi, (4S)-4, 1 l-diethyi-4-hydroxy-9-[(4- piperidinopiperidino)-carbonyloxy]~l H-yrano[3',4',6,7]indoiizino[l,2-bJquinoline-3, 14(4H, 12H)-dione hydrochloride, is commercially available as the injectable solution CAMPT0SAR®. Irinotecan is a derivative of camptothecin which binds, along with its active metabolite 8N-38, to the topoisomerase I-DNA complex. Topotecan HCI, (S)-10-[(dimethylamino)methyl]-4- ethyl-4,9-dihydroxy-lH~pyrano[3',4',6,7]indolizino[l ,2-b]quinoline-3, 14-(4H, 12H)-dione monohydrochloride, is commercially available as the injectable solution HYCAMTIN®.

[00255] Anti-metabolites include (a) purine analogs such as fludarabine, ciadribine, chiorodeoxyadenosine, clofarabine, mercaptopurine, pentostatin, and thioguanine;

(b) pyrimidine analogs such as fluorouracil, gemcitabine, capecitabine, cytarabine, azacitidine, edatrexate, floxuridine, and troxacitabine; (c) antifolates, such as methotrexate, pemetrexed, raltitrexed, and trimetrexate. Anti-metabolites also include thymidylate synthase inhibitors, such as fluorouracil, raltitrexed, capecitabine, floxuridine and pemetrexed; and ribonucleotide reductase inhibitors such as claribine, clofarabine and fludarabine. Antimetabolite neoplastic agents are phase specific anti-neoplastic agents that typically act at S phase (DNA synthesis) of the cell cycle by inhibiting DNA synthesis or by inhibiting purine or pyrimidine base synthesis and thereby limiting DNA synthesis. Consequently, S phase does not proceed and cell death follows, Anti-metabolites, include purine analogs, such as fludarabine, ciadribine,

chiorodeoxyadenosine, clofarabine, mercaptopurine, pentostatin, erythrohydroxynonyladenine, fludarabine phosphate and thioguanine; pyrimidine analogs such as fluorouracil, gemcitabine, capecitabine, cytarabine, azacitidine, edatrexate, floxuridine, and troxacitabine; antifolates, such as methotrexate, pemetrexed, raltitrexed, and trimetrexate. Cytarabine, 4-amino-] -p-D- arabinofuranosyl-2 (I H)-pyrimidinone, is commercially available as CYTOSAR-U® and is commonly known as Ara-C. Mercaptopurine, 1 ,7-dibydro-6H-purine-6-thione monohydrate, is commercially available as PURINETHOL®. Thioguanine, 2-amino-l, 7-dihydro-6H-purine-6- ihione, is commercially available as TABLOID®. Gemcitabiiie, 2'-deoxy-2', 2'-difluorocy†idine monohydrochloride (p-isomer), is commercially available as GEMZAR®.

[00256] Hormonal therapies include (a) androgens such as fluoxymesterone and testolactone; (b) antiandrogens such as bicaluiamide, cvproterone, fSutamide, and nilutamide; (c) aromatase inhibitors such as amiiioglutethimide, anastrozoie, exemestane, formestane, and letrozole;

(d) corticosteroids such as dexamethasone and prednisone; (e) estrogens such as

diethylstilbestrol; (f) antiestrogens such as fulvestrant, raloxifene, tamoxifen, and toremifine;

(g) LHRH agonists and antagonists such as buserelin, goserelin, leuprolide, and triptorelin;

(h) progestins such as medroxyprogesterone acetate and megestrol acetate; and (i) thyroid hormones such as levothyroxine and liothyronine. Hormones and hormonal analogues are useful compounds for treating cancers in which there is a relationship between the hormone(s) and growth and/or lack of growth of the cancer. Examples of hormones and hormonal analogues useful in cancer treatment include, but are not limited to, androgens such as fluoxymesterone and testolactone; antiandrogens such as bicaluiamide, cvproterone, flutamide, and nilutamide; aromatase inhibitors such as amiiioglutethimide, anastrozoie, exemestane, formestane, vorazole, and letrozole; corticosteroids such as dexamethasone, prednisone and prednisolone; estrogens such as diethylstilbestrol; antiestrogens such as fulvestrant, raloxifene, tamoxifen, toremifine, droloxifene, and iodoxyfene, as well as selective estrogen receptor modulators (SERMS) such those described in U.S. Patent Nos. 5,681 ,835, 5,877,219, and 6,207,716; 5a-reductases such as finasteride and dutasteride; gonadotropin-releasing hormone (GnRH) and analogues thereof which stimulate the release of leutinizing hormone (LH) and/or follicle stimulating hormone (FSH), for example LHRH agonists and antagonists such as buserelin, goserelin, leuprolide, and triptorelin; progestins such as medroxyprogesterone acetate and megestrol acetate; and thyroid hormones such as levothyroxine and liothyronine.

[00257] Signal transduction pathway inhibitors are those inhibitors, which block or inhibit a chemical process which evokes an intracellular change, such as cell proliferation or

differentiation. Signal tranduction inhibitors useful in the present invention include, e.g., inhibitors of receptor tyrosine kinases, non-receptor tyrosine kinases, 8H2/8H3 domain blockers, serine/threonine kinases, phosphotidyl inositol-3 kinases, myo-inositol signaling, and Ras oncogenes. [00258] Molecular targeted agents include (a) receptor tyrosine kinase ('RTK') inhibitors, such as inhibitors of EGFR, including erlotinib, gefitinib, and neratinib; inhibitors of VEGFR including vandetanib, semaxinib, and cediranih; and inhibitors of PDGFR; further included are RTK inhibitors that act at multiple receptor sites such as lapatinib, which inhibits both EGFR and HER2, as well as those inhibitors that act at each of C-kit, PDGFR and VEGFR. including but not limited to axitinib, sunitinib, sorafenib and toceranib; also included are inhibitors of BCR-ABL, c-kit and PDGFR, such as imatinib; (b) FKBP binding agents, such as an immunosuppressive macrolide antibiotic, including bafilomycin, rapamycin (sirolimus) and everoliraus; (c) gene therapy agents, antisense therapy agents, and gene expression modulators such as the retinoids and rexinoids, e.g, adapalene, bexarotene, trans-retinoic acid, 9-cis-retinoic acid, and N-(4-hydroxyphenyl)retinamide; (d) phenotype-directed therapy agents, including monoclonal antibodies such as alerntuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab; (e) immunotoxins such as gemtuzumab ozogamicin;

(f) radioimmunocoiijugates such as 1311-tositumomab: and (g) cancer vaccines,

[00259] Several protein tyrosine kinases catalyse the phosphorylation of specific tyrosyl residues in various proteins involved in the regulation of ceil growth, Such protein tyrosine kinases can be broadly classified as receptor or non-receptor kinases. Receptor tyrosine kinases are transmembrane proteins having an extracellular ligand binding domain, a transmembrane domain, and a tyrosine kinase domain, Receptor tyrosine kinases are involved in the regulation of cell growth and are sometimes termed growth factor receptors.

[00260] Inappropriate or uncontrolled activation of many of these kinases, for example by over-expression or mutation, has been shown to result in uncontrolled cell growth. Accordingly the aberrant activity of such kinases has been linked to malignant tissue growth. Consequently, inhibitors of such kinases could provide cancer treatment methods.

[00261] Growth factor receptors include, for example, epidermal growth factor receptor (EGFr), platelet derived growth factor receptor (PDGFr), erbB2, erbB4, vascular endothelial growth factor receptor (VEGFr), tyrosine kinase with immimogiobuiin-like and epidermal growth factor homology domains (Tl E-2), insulin growth factor -I (1GF1) receptor, macrophage colony stimulating factor (cfms), BTK, ckit, cmet, fibroblast growth factor (FGF) receptors, Trk receptors (TrkA, TrkB, and TrkC), ephrin (eph) receptors, and the RET protooncogene.

[ 00262] Several inhibitors of growth receptors are under development and include ligand antagonists, antibodies, tyrosine kinase inhibitors and anti-sense oligonucleotides. Growth factor receptors and agents that inhibit growth factor receptor function are described, for instance, in ath, John C, Exp. Opin. Ther. Patents (2000) 1 0(6):803-81 8; Shawver et al,, Drug Discov. Today (1997), 2(2): 50-63; and Lofts, F. J. et al., "Growth factor receptors as targets", New Molecular Targets for Cancer Chemotherapy, ed. Workman, Paul and Kerr, David, CRC press 1994, London. Specific examples of receptor tyrosine kinase inhibitors include, but are not limited to, sunitinib, erlotinib, gefitfnib, and imatinib.

[00263] Tyrosine kinases which are not growth factor receptor kinases are termed nonreceptor tyrosine kinases. Non-receptor tyrosine kinases useful in the present invention, which are targets or potential targets of anti-cancer drugs, include cSrc, Lck, Fyn, Yes, Jak, cAbl, FAK (Focal adhesion kinase), Brutons tyrosine kinase, and Bcr-Abl, Such non-receptor kinases and agents which inhibit non-receptor tyrosine kinase function are described in Sinh, S. and Corey, S.J., J. Hematotherapy & Stem Ceil Res. ( 1999) 8(5): 465 - 80; and Bolen, J.B., Brugge, J.S., Annual Review of Immunology. (1997) 15: 371-404.

[00264] SH2/SH3 domain blockers are agents that disrupt SH2 or SH3 domain binding in a variety of enzymes or adaptor proteins including, PI3-K p85 subunit, Src family kinases, adaptor molecules (She, Crk, Nek, Grb2) and Ras-GAP. SH2/SH3 domains as targets for anti-cancer drugs are discussed in Smithgail, I.E., J. Pharmacol. Toxicol. Methods. (1995), 34(3): 125-32. Inhibitors of Serine/Threonine Kinases including MAP kinase cascade blockers which include blockers of Raf kinases (rafk), Mitogen or Extracellular Regulated Kinase (MEKs), and

Extracellular Regulated Kinases (ERKs); and Protein kinase C family member blockers including blockers of PKCs (alpha, beta, gamma, epsilon, mu, lambda, iota, zeta). IkB kinase family (IKKa, IKKb), PKB family kinases, AKT kinase family members, and TGF beta receptor kinases. Such Serine/Threonine kinases and inhibitors thereof are described in Yamamoto, T., Taya, 8., Kaibuchi, K„ J. Biochemistry. (1999) 126 (5): 799-803; Brodt, P, Samani, A, & Navab, R, Biochem. Pharmacol (2000) 60: 1 101 -1 107; Massague, J ., Weis-Garcia, F., Cancer Sun'. (1996) 27:41-64; Philip, P.A, and Harris, AL, Cancer Treat. Res. (1995) 78: 3-27; Lackey, K. et al. Bioorg. Med. Chern. Letters, (2000) 10(3): 223-226; U.S. Patent No. 6,268,391 ; and Martinez-Lacaci, L, et al, Int. J. Cancer (2000), 88(1): 44-52. Inhibitors of Phosphotidyi inositol-3 Kinase family members including blockers of P13- kinase, ATM, DN A-PK, and Ku are also useful in the present invention. Such kinases are discussed in Abraham, RT. Current Opin. Immunol. (1996), 8(3): 412-8; Canman, C.E., Lim, D.S., Oncogene (1998) 17(25): 3301 - 8; Jackson, S.P., Int. J. Biochem. Cell Biol. (1997) 29(7):935-8; and Zhong, H. et al, Cancer Res. (2000) 60(6): 1541 -5. Also useful in the present invention are Myo-inositol signaling inhibitors such as phospholipase C blockers and Myoinositol analogues. Such signal inhibitors are described in Powis, G., and Kozikowski A, (1994) New Molecular Targets for Cancer Chemotherapy, ed., Paul Workman and David Kerr, CRC Press 1994, London.

[00265] Another group of signal transduction pathway inhibitors are inhibitors of Ras Oncogene. Such inhibitors include inhibitors of farnesyltransferase, geranyl-geranyl transferase, and CAAX proteases as well as anti-sense oligonucleotides, ribozymes and immunotherapy. Such inhibitors have been shown to block ras activation in cells containing wild type mutant ras , thereby acting as antiproliferation agents. Ras oncogene inhibition is discussed in Scharovskv, O.G., Rozados, V.R, Gervasoni, SI, Matar, P., J. Biomed. Sci. (2000) 7(4): 292-8; Ashby, M.N., Can: Opin. Lipidol. (1998) 9(2): 99 - 102; and Oliff, A., Biochim. Biophys. Acta, (1999)

1423(3):C 19-30.

[00266] As mentioned above, antibody antagonists to receptor kinase ligand binding may also serve as signal transduction inhibitors. This group of signal transduction pathway inhibitors includes the use of humanized antibodies to the extracellular ligand binding domain of receptor tyrosine kinases. For example Imclone C225 EGFR specific antibody (see Green, M.C. et al., Cancer Treat. Rev.,, (2000) 26(4): 269-286); Herceptin® erbB2 antibody (see Stern, DF, Breast Cancer Res. (2000) 2(3): 176-183); and 2CB VEGFR2 specific antibody (see Brekken, R.A. et al, Cancer Res. (2000) 60(18):5117-24).

[00267] Non-receptor kinase angiogenesis inhibitors may also find use in the present invention. Inhibitors of angiogenesis related VEGFR and ΉΕ2 are discussed above in regard to signal transduction inhibitors (both receptors are receptor tyrosine kinases). Angiogenesis in genera] is linked to erhB2/EGFR. signaling since inhibitors of erbB2 and EGFR. have been shown to inhibit angiogenesis, primarily VEGF expression. Thus, the combination of an erbB2/EGFR inhibitor with an inhibitor of angiogenesis makes sense. Accordingly, nonreceptor tyrosine kinase inhibitors may be used in combination with the EGFR/erbB2 inhibitors of the present invention. For example, anti-VEGF antibodies, which do not recognize VEGFR (the receptor tyrosine kinase), but bind to the ligand; small molecule inhibitors of integrin (alphav beta3) that will inhibit angiogenesis; endostatin and angiostatin (iion-RTK) may also prove useful in combination with the disclosed erb family inhibitors. (See Brans, CJ et al., Cancer Res. (2000), 60(1 1): 2926-2935; Schreiber AB, Winkler ME, & Derynck R., Science (1986) 232(4755): 1250-53; Yen L. et al, Oncogene (2000) 19(31): 3460-9).

[00268] Agents used in immunotherapeutic regimens may also be useful in combination with the compounds of formula (I). There are a number of immunologic strategies to generate an immune response against erbB2 or EGFR. These strategies are generally in the realm of tumor vaccinations. The efficacy of immunologic approaches may he greatly enhanced through combined inhibition of erhB2/EGFR signaling pathways using a small molecule inhibitor.

Discussion of the immunologic/tumor vaccine approach against erbB2/EGFR are found in Reilly RT, et al, Cancer Res. (2000) 60(13):3569-76; and Chen Y, et al., Cancer Res. (1998)

58(9): 1965-71.

[00269] Agents used in pro-apoptotic regimens (e.g., bcl-2 antisense oligonucleotides) may also be used in the combination of the present invention. Members of the Bci-2 family of proteins block apoptosis. Upregulation of bcl-2 has therefore been linked to chemoresistance. Studies have shown that the epidermal growth factor (EGF) stimulates anti-apoptotic members of the bci-2 family. Therefore, strategies designed to downregulate the expression of bcl-2 in tumors have demonstrated clinical benefit and are now in Phase Will trials, namely Genta's G3139 bcl-2 antisense oligonucleotide, Such pro-apoptotic strategies using the antisense oligonucleotide strategy for bcl-2 are discussed in Waters JS, et al., J. Clin. Oncol. (2000) 18(9): 1812-23; and Kitada S, et al. Antisense Res. Dev. (1994) 4(2): 71-9.

[00270] Cell cycle signalling inhibitors inhibit molecules involved in the control of the cell cycle. A family of protein kinases called cyclin dependent kinases (CDKs) and their interaction with a family of proteins termed cyclins controls progression through the eukaryotic cell cycle. The coordinate activation and inactivation of different cyclin/CDK complexes is necessary for normal progression through the cell cycle. Several inhibitors of cell cycle signalling are under development. For instance, examples of cyclin dependent kinases, including CDK2, CDK4, and CDK.6 and inhibitors for the same are described in, for instance, RosaniaGR & Chang Y -T., Exp. Opin. Ther. Patents (2000) 10(2):215-30.

[00271] Other molecular targeted agents include FKBP binding agents, such as the immunosuppressive macrolide antibiotic, rapamycin; gene therapy agents, antisense therapy agents, and gene expression modulators such as the retinoids and rexinoids, e.g. adapaiene, bexarotene, trans-retinoic acid, 9-cisretitioic acid, and N-(4 hydroxyphenyl)retmamide;

phenotype-directed therapy agents, including: monoclonal antibodies such as alemtuzumab. bevacizumab, cetuximab, ibritumomab tiuxeian, rifuximab, and trastuzumab; immunotoxins such as gemtuzumab ozogamicin, radioimmunoconjugates such as 131-tositumomab; and cancer vaccines.

[00272] Anti-tumor antibiotics include (a) anthracyc lines such as daunorubicin (including liposomal daunorubicin), doxorubicin (including liposomal doxorubicin), epirubicin, idarubicin, and valrubicin; (b) streptomyces-related agents such as bleomycin, actinomycin, mithramycin, mitomycin, porfiromycin; and (c) anthracenediones, such as mitoxantrone and pixantrone. Anthracyclines have three mechanisms of action: intercalating between base pairs of the DNA/RNA strand; inhibiting topoiosomera.se II enzyme; and creating iron-mediated free oxygen radicals that damage the D A and cell membranes. Anthracyclines are generally characterized as topoisomera.se II inhibitors.

[00273] Monoclonal antibodies include, but are not limited to, murine, chimeric, or partial or fully humanized monoclonal antibodies. Such therapeutic antibodies include, but are not limited to antibodies directed to tumor or cancer antigens either on the cell surface or inside the cell. Such therapeutic antibodies also include, but are not limited to antibodies directed to targets or pathways directly or indirectly associated with CK2. Therapeutic antibodies may further include, but are not limited to antibodies directed to targets or pathways that directly interact with targets or pathways associated with the compounds of the present invention. In one variation, therapeutic antibodies include, but are not limited to anticancer agents such as Abagovomab, Adecatumumab, Afutuzumab, Alacizumab pegol, Alemtuzumab, Altumomab pentetate, Anatumomab mafenatox, Apolizumab, Bavituximab, Belimumab, Bevacizumab, Bivatuzumab mertansine, Blinatumomab, Brentuximab vedotin, Cantuzumab mertansine, Catumaxomab, Cetuximab, Citatuzumab bogatox, Cixutumumab, Ciivatuzumab tetraxetan, Conatumumab, Dacetuzumab, Detumomah, Ecromeximab, Edrecolomab, Elotuzumab,

Epratuzumab, Ertumaxomab, Etaracizumab, Farletuzumab, Figitumumab, Fresolimumab, Galiximab, Glembatumumab vedotin, Ibritumomab tiuxetan, Intetumumab, Inotuzumab ozogamicin, Ipilimumab, iratumumab, Labetuzumab, Lexatumumab, Lintuzumab,

Lucatumumab, Lumiliximab, Mapatumumab, Matuzumab, Milatuzumab, Mitumomab,

Nacolomab tafenatox, Naptumomab esiafenatox, Necitumumab, Nimoiuzumab, Ofatumumab, Olaratuniab, Opoxtuzumab monatox, Oregovomab, Panitumumab, Pemtumomab, Pertuzumab, Pintumomab, Pritumumab, Ramucirumab, Riiotumumab, Rituximab, Robatumumab.

Sibrotuzumab, Tacatuzumab tetraxetan, Tapiitumomab paptox, Tenatumomab, Ticiiimumab, Tigatuzumab, Tositumomab. Trastuzumab, Tremeiimumab, Tucotuzumab celmoleukin, Veltuzumab, Volociximab, Votumumab, Zalutumumab, and Zanolimumab. In some

embodiments, such therapeutic antibodies include, alemtuzumab, bevacizumab, cetuximab, daclizumab, gemtuzumab, ibritumomab tiuxetan, pantiiumumab, rituximab, tositumomab, and trastuzumab; in other embodiments, such monoclonal antibodies include alemtuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab; alternately, such antibodies include daclizumab, gemtuzumab, and pantiiumumab. in yet another embodiment, therapeutic antibodies useful in the treatment of infections include but are not limited to

Afelimoniab, Efungumab, Exbiviamiab, Felvizumab, Foraviraniab, Ibalizumab, Libiviramab, Motavizumab, Nehacumab, Pagibaximab, Palivizumab, Panobacumab, Rafivirumab,

Raxibacumab, Regavirumab, Sevirumab, Tefibazumab, Tuvirumab, and Urtoxazumab. In a further embodiment, therapeutic antibodies can be useful in the treatment of inflammation and/or autoimmune disorders, including, but are not limited to, Adalimumab, Atlizumab, Atorolimumab, Aselizumab, Bapineuzumab, Basiliximab, Benralizimiab, Bertiiimumab, Besilesoniab, Briakinumab, Canakinumab, Cedelizumab, Certolizumab pegoi, Cienoliximab, Daclizumab, Denosumab, Eculizumab, Edobacomab, Efalizumab, Erlizumab, Fezakinumab, Fontolizumah, Fresolimumab, Gantenerumab, Gavilimomab, Golimumab, Gomiliximab, Infliximab, Inolimomab, eliximab, Lebrikizumab, Lerdelimumab, Mepolizumab,

Meteii.rn.umab, Muromonab-CD3, Natalizumab, Ocrelizumab, Odulimomab, Omalizumab, Otelixizuniab, Pascolizumab, Priliximab, Reslizumab, Rituximab, Rontalizumab, Rovelizumab, Ruplizumab, Sifalimumab, Siplizumab, Solanezumab, Stamulumab, Talizumab, Tanezumab, Teplizumab, Tocilizumab, Toralizumab, Ustekinumab, Vedolizumab, Vepalimomab,

Visiiizumab, Zanoiimumab, and Zoiimomab aritox. In yet another embodiment, such therapeutic antibodies include, but are not limited to adalimumab, basiliximab, certolizumab pegol, eculizumab, efalizumab, infliximab, muromonab-CD3, natalizumab, and omalizumab. Alternately the therapeutic antibody can include abciximab or ranibizumab. Generally a therapeutic antibody is non-conjugated, or is conjugated with a radionuclide, cytokine, toxin, drug-acti ating enzyme or a drug-filled liposome.

[00274] Akt inhibitors include lL6-Hydroxymethyl-chiro-inositol-2-(R)-2-0-methyl-3-0- oct.adecyl.-ii/?-glycerocarbonate, SH-5 (Calbiochem Cat. No, 124008), SH-6 (Calbiochem Cat. No. Cat. No. 124009), Calbiochem Cat. No. 12401 1 , Triciribine (NSC 154020, Calbiochem Cat. No. 124012), 10-( '-(N-diethylamino)buty l)-2-chloropb.enoxazine, Cu(II)Cl?(3-Formylchromone thiosemicarbazone), l,3-dihydro-l-(l -((4-(6-phenyl-l Fl-imidazo[4,5-g]quinoxalin- 7-yl)phenyl)methyi)-4-piperidiiiyl)~2H ienzimidazol- GSK690693 (4-(2-(4-amino-l ,2,5- oxadi.azol-3 ~yl)~ 1 -ethyl-7- { [(3 S)~3 -piperidinylmethyljoxy } - 1 H-imidazo[4,5-c]pyridin-4-yl)-2- methyl-3-butyn-2-ol), SRI 3668 ((2,10-dicarbethoxy-6-methoxy-5,7-dihydro-indolo[2,3-b] carbazole), GSK2141795, Perifosine, GSK2.1 1 10183, XL418, XL147, PF-04691502, BEZ-235 [2-Methyl-2-[4-(3-methyl-2-oxo-8-quinolin-3-yl-2,3-dihydro-imidazo[4,5-c]quinolin-l-yl)- phenylj-propionitrile], PX-866 ((acetic acid (1S,4E,10R,1 lR,13S,14R)-[4- dialIylammomethylene-6-hydroxy-l-methoxymethyl-l 0,13-dimethyi-3,7,17-trioxo- 1 ,3,4,7,10,11,12,13,14,15,16,17-dodecahydro-2-oxa-cyclopenta[a]phenanthren- 11 -yl ester)), D- 106669, CAL- 101, GDC0941 (2-( 1 H-indazol-4-yl)-6-(4-methanesulfonyI-piperazin- 1 - ylme l)-4-motpholin^-y hieno[3,2-d]pyrimidine), SF1126, SF1 188, SF2523, TG lOO-1 15 [3-[2,4-diamino-6-(3-hydroxyphenyi)pteridin-7-yl]phenol]. A number of these inhibitors, such as, for example, BEZ-235, PX-866, D 106669, CAL-101, GDC0941, SF1126, SF2523 are also identified in the art as PI3K/mTOR inhibitors; additional examples, such as Pl-103 [3-[4-(4- morpholinylpyrido[3^2 ,5]furo[3,2-d]pyTimidin-2-yl]phenol hydrochloride] are well-known to those of skill in the art. Additional well-known PI3K inhibitors include LY294002 [2-(4- morpho[myl)-8-phenyl-4H-l-benzopyran-4-one] and wortmannin. mTOR inhibitors known to those of skill in the art include temsirolimus, deforolimus, sirolimus, everolimus, zotarolirnus, and biolimus A9. A representative subset of such inhibitors includes temsirolimus, deforolimus, zotarolirnus, and biolimus A9,

[00275] HDAC inhibitors include (i) hydroxamic acids such as Trichostatin A, vorinostat (suberoylanilide hydroxamic acid (SAHA)), panobinostat (LBH589) and belinostat (PXD101) (it) cyclic peptides, such as trapoxin B, and depsipeptides, such as romidepsin (NSC 630176), (iii) benzamid.es, such as MS-275 (3-pyridylmethyl-N-{4~[(2~amiiiopheiiyl)-carbamoyl]- benzyl} -carbamate), CI994 (4-acetylamino- -(2aminophenyl)-benzamide) and MGCD0103 (N- (2-aminophenyl)-4-((4-(pyridin-3-yl)pyrimidin-2-ylamino)methyl)be (iv) electrophilic ketones, (v) the aliphatic acid compounds such as phenylbutyrate and valproic acid.

[00276] Hsp90 inhibitors include benzoquinone ansamycins such as geldanamycin,

17-DMAG ( 17-Dimethylamino-ethy lamino- 17-demethoxygeldanamycin), tanespimycin

(17-AAG, 17-allylaniino- 17-demethoxygeldanamycin), ECS, retaspimycin (IPI-504,

18,21 -didehydro-17-demethoxy- 18,21 -dideoxo- 18,21 -dihydroxy- 17-(2-propeny3 amino)- geldanamycin), and herbimycin; pyrazoles such as CCT 018159 (4-[4-(2,3-dihydro- Ϊ ,4- benzodioxm-6-yl)-5-methyl-lH-pyrazol-3-yl]-6-ethyl-l,3-benzenediol); macrolides, such as radicocol; as well as BHB021 (CNF2024), SNX-5422, STA-9090, and AUY922.

[00277] Miscellaneous agents include altretamine, arsenic trioxide, gallium nitrate, hydroxyurea, levamisole, mitotane, octreotide, procarbazine, suramin, thalidomide,

lenalidomide, photodynamic compounds such as methoxsalen and sodium porfimer, and proteasome inhibitors such as bortezomib.

[00278] Biologic therapy agents include: interferons such as interferon-a2a and interferon- a2b, and interieukins such as aldesleukin, denileukin diftitox, and oprelvekin. [00279] In addition to these anticancer agents intended to act against cancer cells, combination therapies including the use of protective or adjunctive agents, including:

cytoprotective agents such as arrnifostine, dexrazonxane, and rnesna, phosphonates such parmidronate and zoledronic acid, and stimulating factors such as epoetin, darbepoetin, filgrastim, PEG-filgrastim, and sargramostim, are also envisioned.

Examples:

[00280 J The following examples illustrate and do not limit the invention. Example 1. Synthesis or5-diR>ropy; a/oR>j 1.5-a ipyr;m:din-7~aminc

[00281] To the reaction flask, 5,7-dichloropyrazolo[l ,5-a]pyrimidine (3 g, 16 mmol) was added along with ammonium hydroxide solution (48 niL). The heterogeneous reaction was reiluxed at 85°C for 12 hours. After cooling to room temperature, the mixture was filtered, washed with water, and dried under vacuum overnight. The product, 5-chloropyrazolo[l ,5- a]pyrimidin-7-amine, was collected as an off-white solid in 88% yield. LCMS (M+l=169)

Example 2. Synthesis of tert-hutyl 5-chl.oropyrazoloP ,5-a]pyrimidin-7-ylcarbamate

E82] To the reaction flask, 5-chloropyrazolo[l ,5-a]pyrimidiri-7-amme (2.4 g, 14.1 mmol) was added to dichloromethane (35 ml.,) along with di-tert-butyl dicarbonate (3.7 g, 17 mmol), triethylamine (2.4 mL, 17 mmol) and DMAP (100 nig, 0.8 mmol). The reaction was stirred at room temperature for 6 hours then diluted with DCM, washed with saturated NaHC0₃ solution (3x) followed by washing with brine. The organic layer was isolated, dried over anhydrous gS0₄, filtered, and evaporated to dryness. The product, tert-hutyl 5-chloropyrazolo[l ,5- a]pyrimidm-7-yicarbamate, was collected as an off-white solid in 98% yield. LCMS (M-t-Butyl -213) Example 3.

[00283] To tert-butyl 5-chloropyrazolo[l,5-a]pyriimdin-7-yIcarbamate (3.7 g, 13.8 mmol) in DMF (36 mL), POCI₃ (7,7 mL, 82.9 mmol) was added dropwise at 0°C. After the addition was complete, the reaction was allowed to warm to room temperature and stirred for 8 hours. Then, the reaction was quenched by slow addition to ice cold 6N NaOH, The mixture was diluted with water then the solid was collected by filtration. The solid was washed several more times with water and dried under vacuum overnight. The product, tert-butyl 5-chloro-3- formylpyrazolo[l ,5-a]pyrimidin-7-yScarbamate, was collected as a solid in 27% yield. The product did not ionize on LCMS unless first deprotected using TFA/DCM (1 : 1). LCMS (M+l=197)

Example 4. Synthesis of 7-ammo-5-(3-chlorophenylaniino)pyrazolo[ 1 ,5~a]pyrimidine-3- carbaldehyde

E84] Tert-butyl 5-chloro-3-formylpyrazolo[ 1 ,5-a]pyrimidin-7-ylcarbamate (1.1 g, 3.8 mmol) w^ras added to 1 ,4-dioxane ( 15 mL) along with 3-chloroamline (2.4 mL, 22.6 mmol) and p-toluenes ulfonic acid moiiohydrate (73 mg, 0.4 mmol). The reaction was heated at 95°C for 12 hours then cooled to room temperature, diluted with water and filtered. Analysis of the recovered solid by LCMS showed product mass (M+l = 288), as w^rell as, product with chloro aniline imine mass (M+l = 397), To completely convert this mixture to the desired product, the solid was dissolved in 6 mL of MeOH/conc. HC1 solution (1 : 1) and heated at 60°C for 1 ,5 hours. The reaction was quenched by slow addition to ice cold 6N NaOH. The mixture was diluted with water then the solid was collected by filtration. The solid was washed several more times with water then dried under vacuum overnight. The product, 7-amino-5-(3- chlorophenylamino)pyrazolo[l,5-a]pyrimidine-3-carbaldehyde, was collected as orange-red solid in 38% yield. LCMS (M+l=288) Example 5. S nihcsis t if 5-(( 7-an=i:ic)-3-(3-chior piicn ;n :irju)ps raxt 1.5- jpy; si idin- - yr)meth lene^")imidazolidine-2,4-dione

[00285] To the reaction vial, 7-amino-5-(3-chlorophenylamino)pyrazolo[l ,5-a]pyrimidine-3- carbaidehyde (411 mg, 1.4 mmol) was added to ethanol (5.2 mL) along with hydantoin (143 mg, 1.4 mmol) and piperidine (141 μ,Ε, 1.4 mmol). The reaction was heated at 80°C for 60 minutes in the microwave. The reaction was then cooled to room temperature and diluted with water. The solid was collected by filtration, washed with water and cold ethanol. The materia] was dried under vacuum overnight. The product, 5-((7-amino-5-(3- chlorophenyiamino)pyrazolo[! ,5-a]pyrm was recovered as a red solid in 54% yield. LCMS (M+l = 370)

Example 6. Synthesis of N-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-4- viidene methyl)pyrazol [^'L5-a]pyrimidm-7-yl piperidine-4-carboxarn.ide

[00286] To the reaction via], 5-((7~amino-5~(3-chiorophenylammo)pyrazolo[l ,5-a]pyrimidin- 3-yl)methylene)imidazolidine-2,4-dione (15 mg, 0.04 mmol) was added to DMF (0.2 mL) along with HBTU (30 mg, 0.08 mmol), DIEA (28 μΕ, 0.16 mmol) and 1 -(tert- butoxycarbonyl)piperidine-4-carboxylic acid (18 mg, 0.08 mmol). The reaction was stirred at room temperature for 8 hours then heated at 95°C for 4 hours. The reaction was then cooled to room temperature and diluted with water. The solid was collected by filtration, washed with water, IN HCl solution, and more water. The material was then dissolved in 5% DCM/^'MeOH and purified by prep HPLC. The isolated fractions were combined and evaporated to dryness. The material was dissolved in 1 mL of TFA/DCM (1 :1) and stirred at room temperature for 1 hour. The solvent was removed by evaporation under a stream of nitrogen and the crude material was washed with IN NaOH followed by water. The solid was collected by filtration and dried under vacuum overnight. The product, N-(5-(3-chlorophenylamino)-3-((2,5- dioxoimidazolidin~4-ylidene)methyl)pyrazoio[l,5-a]pyrimidin-7-yl)pipeiidm

was recovered as a solid in 2% yield. LCMS (M · i = 481).

[00287] Table 1 below shows the biological acvities of Examples 5 and 6 as listed as Compounds Al and Bl .

Table 1.

Example 7. Synthesis of 5-chloro-N-c clopropyipyr3zolo[l,5-a]pyrhnidin-7-amine

E88] To 5,7-dichloropyrazolo[l,5-a]pyrimidine (200 mg, 1.06 mmol) in ACN was added Et₃N (148 μΐ, 1.06 mmol) and cyclopropylamme (75 μΐ, 1.06 mmol). The reaction was reiluxed at 80°C overnight. The mixture was concentrated under reduced pressure, dissolved in DCM, and washed with water. The resulting organic layer was dried over Na₂S0₄ and concentrated under reduced pressure to afford 156 mg of 5-chloro-N-cyclopropy lpyrazolo[ 1 ,5-a]pyrimidin-7- amine (70% yield). LCMS (M+l=209)

Example Synthesis of 5-chloro-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidine-3- carbaldehyde

[00289] To 5-chloro-N-cyc3.opropylpyrazolo[l ,5-a]pyTimidin-7-amine (156 mg, 0.75 mmol) in DMF^' was added POCI3 (205 μΐ, 2,25 mmol). The mixture was stirred at room temperature for 3 hours. Ice was added to quench excess POCI₃ then the mixture was neutralized with 1 NaOH. DCM was added and the product was extracted three times. The organic layer was dried over Na₂S0₄ and concentrated under reduced pressure to yield 5-chloro-7- (cyclopropylamino)pyrazolo[l,5-a]pyrimidine-3-carbaldehyde. Some residual DMF could not be removed. LCMS (M I 23 ?)

Example 9. Synthesis of 5-(3-chlorophenylarmno)-7-(cyclopropylamino)pyrazolo[ 1 ,5- a]pyrimidine-3-carbaldehyde

[00290] To 5 -chloro-7-(cyclopropylamino)pyrazolo [1,5 -a]pyrimidine-3 -carbaldehy de (177 mg, 0.75 mmol) in 1,4-dioxane was added 3-chloroaniline (397 μΙ, 3.75 mmol). The mixture was heated in microwave at 120°C for 60 minutes. The precipitate was filtered off, and the filtrate was purified by prep TLC (l%MeOH/DC ) to yield 26 mg ( 1 1% yield) of 5-(3- chlorophenylaniino)-7-(cyclopropylamino)pyrazolo[l,5-a]pyrimidine-3-carbaldehyde, LCMS

(M+ 1=328)

Example 10. Synthesis of 5-((5-(3-chlorophenylamino)- a]pyTimidin-3- l)methylene)imidazolidine-2,4-dione

[002 1 ] To -(3 -chloropheny lamino)-7~(cyclopropylamino)pyrazol 0 [1 ,5 -a]pyrirmdine-3 - carbaldehyde (26 mg, 0.08 mmol) in EtOH was added hydantoin (8 mg, 0.08 mmol) and piperidiiie (8 μΐ. 0.08 mmol). The mixture was stirred at 70°C over the weekend. Insolubilities were filtered off, and filtrate was concentrated under reduced pressure. Filtrate was then dissolved in MeOH and isolated by prep HPLC to yield 5-((5-(3-chlorophenylamino)-7- (cyclopropylaniino)pyrazolo[l,5-a]pyrimidin-3-yl)methylene)imi LCMS ( · i 410)

Example 11. Synthesis of 7-{cyck^propyja.niino)-5-{3-fl-Uorophen

a]pyrimidine-3-carbaldehyde

[00292] 7 e/ -butyl 5-chloro-3-formylpyrazoio[ 1 ,5- ]pyrimidin-7-yl(cyclopropyl)carbamate (0.2 g, 0,59 mmol) was suspended in ethanol (2 mL). 3-fSuoroaniline (189 mg, 1.48 mmol) was added, followed by 4M HCl/dioxane (0.3 mL, 1.18 mmoi). The reaction was heated to 80 °C for 6 h, and then the volatiles were removed in vacuo. The residue was diluted with water (10 mL) and the pH was adjusted to 12 by the addition of 6M NaOH. The solution was stirred for 0.5 h, then the precipitate, which is a mixture of fcrf-butyl cyclopropyl(5-(3-fluorophenylamino)-3- formylpyrazo!o[l ,5-ci]pyrimidin-7-y!)carbamate and the corresponding imine, were isolated by filtration and dried in vacuo. The imine was hydrolyzed by dissolving in methanol (9 mL), 1,4- dioxane (3.6 mL) and 6M HCl (9 mL) and heating at 60 °C for 5 h. The solution was poured onto ice (50 mL) and the pH was adjusted to 12 by addition of 6M NaOH. The precipitate was isolated by filtration and dried in vacuo to provide tert-bvXyl cyclopropyl(5-(3- fluorophenylamino)-3-fonnylpyrazolo[i ,5-a]p>T-imidin-7-yl)carbamate (172 mg, 93%). LCMS ( · i 12) Example 12. Synthesis of 5 -((7-(cyclopropylarnin o)-5 -(3 -fiuoropheny 1 aminojpyraz' ,5:

a]pyrimidin-3- l)methylene)imidazolidine-2,4-dione

[00293] Hydantoin (69 mg, 0.69 mmol) and piperidine (69 .uL, 0,69 mrnol) were added to 7- (cyclopropylaniino)-5 -(3 -fluorophenylamino)pyrazolo [ 1 ,5 -a]pyrimidine-3 -carbaldehyde (72 mg, 0.23 mrnol) dissolved in ethanol (1.1 mL). The reaction was heated at 80 °C. After 15 h, the reaction was cooled to r.t., diluted with water (5 mL), and the precipitate was collected and washed with 1 : 1 ethanohwater (5 mL). The bright yellow solid was dried in vacuo to give (Z)- 5-((7-(cyclopropylamino)-5-(3-fluorophenylamino)pyrazolo[l,5-a]pyrimidm-3- yl)methylene)imidazolidine-2,4-dione (25 mg, 10% over 3 steps). LCMS (M+ 1=507)

[00294J The compounds described in the following Table 2A were prepared using chemistries similar to those exemplified in Example 11 and Example 12. All compounds were

characterized by LCMS, Table 2B shows the biological activities of the compounds listed in Table 2A.

Table 2A.

Table 2B.

Example 13. S nihcsis of 7-(cy cloprop l.amino)- -(3 ,5 -difi uoropheriylamino) yrazolo [ 1 ,5 - a]pyrimidine-3-carbaldehyde

[00295] 3,5-DifIuoroaniline (29 mg, 0.22 mmol), Cs₂C0₃ (67 mg, 0.21 mmol) were added to Tert-butyl 5-chloro-3-formylpyrazolo[l,5- ]pyrimidin-7-yl(cyclopropyl)carbamate (50 mg, 0.15 mmol) dissolved in 1 ,4-dioxane (1 mL). Racemic BINAP (6 mg, 0.06 mmol) and palladium! Π ) acetate (4 mg, 0.04 mmol) were then added. The mixture was sealed and iiTadiated at 110 °C for 20 min in the microwave. Et₂0 (3 mL) was added and the solution was filtered. The filtrate was concentrated in vacuo. The crude residue was dissolved in dichloromethane (1.5 mL) and trifluoroacetic acid (1.5 mL). A fter 1 h, the solution was concentrated under a stream of air. The residue was triturated with 20% 2-propanol/hexanes. The product was filtered to yield 7- (cy clopropylamino)-5 -(3 ,5 -di fl uorophenylarmiio)pyrazolo [ 1 ,5 -a]pyrimidine-3 -carbaldehyde (39 mg, 80%). LCMS i^'M 1 330)

[00296] The compounds described in the following Table 3 were prepared using chemistries similar to those exemplified in Example 13. All compounds were characterized by LCMS.

Table 3.

Example 14. Synthesis of (Z)-5-((7-(cyclopropylamino)-5-(3,5- difluorophenyjammo)pyrazolo[l,5- ]pyri

[00297] Hydantoin (28 mg, 0.28 mmol) and piperidine (42 nL, 0,42 mmol) were added to 7-(cyclopropylamino)-5 -(3 ,5 -difluorophenylamino }pyrazo io [ 1 , 5 -a]pyrimidine-3 -car baldehyde (52 mg, 0.16 mmol) dissolved in ethanol (1 mL). The reaction was heated at 80°C. After 12 h, the reaction was cooled to r.t., diluted with water (2 mL), and the precipitate was collected and washed with 1:1 ethanohwater (5 mL). The solid was dried in vacuo to give (Z)-5-((7- (cyclopropylamino)-5-(3,5-difiuorophenylan^'!ino)pyrazolo[l,5-a]pyrin^'!idm-3- yl)methylene)imidazolidine-2,4-dione (18 mg, 28% over 3 steps). LCMS (M+ 1=440)

[00298 J The compounds described in the following tables were prepared using chemistries similar to those exemplified in Example 14. All compounds were characterized by LCMS. Table 4B shows the biological activities of the compounds listed in Table 4A.

Table 4A.

Table 4B.

CK2: IC50 PI 2: 1C50 AB: MDAMB453 AB: BxPC3

(μΜ) (5 μΜ ATP) (μΜ) (μΜ)

11 <0.01 > 2.5000 11.121 15.148

Jl <0.01 2.4799 7.116 3.924

Kl <0.01 > 2.5000 7.711 5.66

LI <0.01 > 2.5000 0.5 1.354

Ml <0.01 > 2.5000

N1 <0.01 1.9706

01 <0.01 > 2.5000

PI <0.01 > 2.5000

Ql <0.01 > 2.5000

Rl <0.01 > 2.5000

SI <0.01 2.041

00299] The compounds described in the following Table 5 were prepared following the general scheme below using chemistries similar to Example 13 and Example 14.

Example 15, Synthesis oi\5-¾3-chloro-4-fluorophem amino)-7-

(cyclopropylmethylamino)pyrazolo[L5-a]pyrimidine-3-carbajdehyde

[00300] To ferf-butyl 5-chloro-3-formylpyrazolo [1, 5-a] pyrimidin-7-yl (cyclopropylmethyl) carbamate (50 mg, 0.14 mmol) in 1 mL of 1 ,4 dioxane was added cesium carbonate (65 mg, 0.2 mmol), Pd (OAc)₂ (4 mg, 0.006 mmol), (+)-BINAP (5 mg, 0.009 mmol), 3-chloro-4- iiuoroanilme (31 mg, 0.21 mmol). The reaction mixture was heated in microwave at 1 10°C for 20 minutes. The mixture was then cooled to room temperature, water was added, and the product was extracted with ether. The organic layer was then concentrated under reduced pressure and the crade product was dissolved in 1 : 1 mixture of dichloromethane and

trifluoroacetic acid at room temperature for Ihour. The reaction mixture was concentrated with 10 mL of dichloromethane. To the reaction mixture, ether/ hexanes (1 :1 ) w^ras added and the flask was sonicated for 10 minutes then filtered to obtain the yellow precipitate. The precipitate was washed with hexane to yield 5-(3~chloro~4-fluorophenylamino)-7- (cyclopropylmethylamino) pvrazolo [1 , 5-a] pyrimidme-3-carbaldehyde. LCMS (M+ 1=460) Example 16. Synthesis of 5 -{{5 -{3 -chloro-4-ίΙ ιιιη ti ho;¾s i;nriir_:t> }-7-

(cyclopropylmethylammo)pyrazolo[l ,5-aJpyrimidin-3-yl)methy

dione

[00301] To 5-(3-chloro-4-fluoropheny[amino)-7-(cyclopropylmethylamino) pyrazoio [1 , 5-a] pyrimidme-3-carbaldehyde (40 mg, 0.09 mmol) in 1 .0 mL of ethanoi was added hydantoin (9 mg, 0.09 mmol) and piperdine (8 ul). The reaction was heated at 80°C overnight, cooled to room temperature, filtered, and washed with ethanoi to yield 20 mg (31% yield) (Z)-5-((5-(3- ch[oro-4-fluoropheny[amino)-7-(cyclopropylmethylamino) pyrazoio [1 , 5-a] pyrimidin-3-yl) methylene) imidazolidine~2, 4-dione as a yellow powder. LCMS (M+l ^::::442)

[00302 J The compounds described in the following tables were prepared using chemistries similar to those exemplified in Example 15 &nd Example 16. All compounds were

characterized by LCMS. Table 6B shows the biological activities of the compounds listed in Table 6A.

Table 6 A.

Table 6B,

8 CK2: IC50 PI 2: 1C50 AB: MDAMB453 AB: BxPC3

(μΜ) (5 μ,Μ ATP) (μΜ) (μΜ)

Ul <0.1 > 2.5000

VI <0.1 > 2.5000 5.086 6.188

Wl <0.1 > 2.5000 7.424 9.207

XI <0.1 > 2.5000 6.935 7.986

Yl <0.1 > 2.5000

7Λ <0.1

A2 <0.1

B2 <0.1

C2 <0.1

D2 <0.1

E2 <0.01

F2 <0.1

G2 <0.01

H2 <0.01

12 <0.1

Example 17. Synthesis o crl-ruil l 5-a/ick>-3-iormvipyra/tsio[ i ,5-a jpvs iiTiiciin-?- yl(cyclopiOpyl)carbamate

[00303J To tert-butyl 5-chloro-3-formylpyrazolo [1,5-a] pyrimidin-7-yl (cyclopropyl) carbamate (500 mg, 1.5 mmol) in dimethylformamide was added sodium azide (150 mg, 2.3 mmol) then the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was then partitioned between ethyl acetate/water. The organic layer was collected, dried over sodium sulfate, filtered, and concentrated under high vacuum to yield tert-butyl 5- azido-3-fomylpyrazolo[l,5-a]pyrimidin-7-yl(cyclopropyl)carbamate. The crude product was taken to next step without further purification. LCMS (M+l ^::::344) Example 18. S nihcsis of 5-mulm)-7-fcyd jm>pyiaminn)pyra/.<y [ j ,5-;; jpyi'liiiiciinc-}- carbaldehyde

[00304] The crude product, tert-butyl 5-azido-3-formylpyrazolo [1 , 5-a] pyrimidin-7-yl (cyclopropyl) carbamate was subjected to hydrogenation using 10% wt palladium on carbon in ethanol. The reaction was stirred under hydrogen for 3 hours. The mixture was filtered through celite and sonicated with 1 : 1 mixture of ethyl acetate and hexane. The yellow solid was filtered and dried under reduced pressure and was dissolved in (1 : 1) DCM/TFA at room temperature for 1 hour. The reaction mixture was washed with aqueous sodium bicarbonate solution and extracted with dichloromethane. The organic layer was concentrated and dried under high vacuum to yield 5-ammo-7-(cyclopropylamino) pyrazolo [1 , 5-a] pyrimidine-3-carbaldehyde as product 310 mg (95% yield on three steps). LCMS ί^' Μ I 2 1 )

Example 19. Synthesis of (Z)-5-((5-amino-7-( cyclopropylamino)pyrazolo j^" 1 ,5-a]pyrimidin-3- yi)methylene)iniidazolidine-2,4-dione

[00305] To 5-amino-7-(cyclopropylammo) pyrazolo [1 , 5-a] pyrimidine-3-carbaldehyde (75 mg, 0.34 mmol) in 1.0 mL ethanol was added hydantoin (34 mg, 0,34 mmol) and piperdine (33 ui). The reaction was heated at 80°C overnight, The reaction mixture was cooled to room temperature and yellow precipitate was filtered, washed with ethanol to yield 45 mg (44% yield)

Z)-5-((5-amino-7-(cyclopropylamino)pyrazolo[l,5-a]pyrimi

2,4-dione. LCMS ( M 1 300 ) Example 20. Synilu:sis pl^'mcthyl 7-f yc:k prc)pyjaniln<>>-3-kn niyipyr;j/C)lo[ i ,5~njpyrimu¾in-5- vicarbamate

To 5-amino-7-(cyclopropylamino) pyrazolo [1, 5 -a] pyrimidine-3-carbaldehyde (50 mg, 0.23 mmol) in 1.0 mL tetrahydrofuran was added methyl chloroforniate (35 ul, 0.46 mmol) and DIEA (39 ul). The reaction mixture was heated at 60°C for one hour. The reaction was partitioned between ethyl acetate /water. The organic layer was collected, dried over sodium sulfate, and concentrated under high vacuum to yield methyl 7-(cyclopropyiamino)-3- formylpyrazolo[l,5-a]pyrimidin-5-ylcarbamate. The crude product was taken to next step without further purification. LCMS (M+ 1=276)

Example 21. Synthesis of (Z)-methyl 7-(cyclopropylamino)-3-((2,5-dioxoiniidazolidin-4-

17] To methyl 7-(cyclopropylamino)-3-formylpyrazolo [1, 5 -a] pyrimidin-5-ylcarbamate (33 mg, 0.12 mmol) in 1.0 mL ethanol was added hydantoin (12 mg, 0.12 mmol) and piperdine (1 1 ul). The reaction was heated at 80°C for two hours. The reaction was cooled to room temperature, die yellow precipitate was filtered, and washed with ethanol to yield 1 mg (40% yield) (Z)-methyl 7-{cyclopropylamino)~3~({2, 5-dioxoimidazolidin-4-ylidene) methyl) pyrazolo [1, 5 -a] pyrimidin-5-ylcarbamate. LCMS (M+l=358) Example 22. Synthesis of N-(7-(cyciopro

yr)cyclopropanecarboxamide

[00308] To 5-amino-7-(cyclopropylamino) pyrazolo [1, 5 -a] pyrimidine-3-carbaldehyde (step b) (58 nig, 0.266 mmol) in 1 .0 mL tetrahydrofuran was added cyclopropane carbonyl chloride (38 ul, 0. 19mmoi) and DIPEA (39.0 ul). The reaction mixture was heated at 60°C for one hour. The reaction was partitioned between ethyl acetate and water, the organic layer was dried under sodium sulfate concentrated on high vac to yield N-(7-(cyclopropylamino)-3- formylpyrazolo [1 , 5-a] pyrimidin-5-yl) cyciopropanecarboxamide. The crude product was taken to next step without further purification. LCMS (M+ 1=286)

Example 23. Synthesis of (Z^')-N-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidm-4-

[00309] To -(7-(cyclopropylamino)-3-formylpyrazo[o [ 1, 5-a] pyrimidin-5-yl)

cyciopropanecarboxamide (74 mg, 0.26 mmol) in 1.0 mL etbanol was added hydantoin (26 mg, 0,26 mmol) and piperdine (24 ul). The reaction was heated at 80°C for two hours. The reaction mixture was cooled to room temperature, concentrated and diluted with MeOH. The produc t was purified by prep HPLC to yield 14 mg (20% yield) (Z)-N-(7-(cyc!opropyiamino)-3-((2,5- dioxoimidazolidin-4-ylidene)methyl)pyrazolo[l,5-a]pyrirrd

LCMS (M+l=368)

[00310] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 20 and Example 16. All compounds were characterized by LCMS, Table 7B shows the biological activities of the compounds listed in Table 7 A.

Table 7A.

Table 7B,

Example 24. Synthesis of -((5 -(chloTO^

yr)methylene^")imidazolidine-2,4-dione

[00311] Tert-hvXyl 5-chloro-3-formylpyrazolo[l,5-aJpyrimidin-7-yl(cyclopropyl)carbamate (0.5 g, 1 .48 mmol) was dissolved in glacial acetic acid (5 niL). NaOAc { 1.21 g, 14.8 mmol) and hydantoin (356 mg, 3.56 mmol) were added and the reaction was placed in a 110 °C bath for 4 d. The solution was cooled to r.t. and water (15 mL) was added. The precipitate was filtered and then triturated with ethanol (5 mL) and then CH₂CI₂ (5 mL) to give (Z)-5-((5-(chloro-7- (cyclopropylamino)pyrazolo[l ,5- ]pyrimidin-3-yi)methylene)imidazolidine-2,4-dione (202 mg, 43%). LCMS (ES): >85% pure, m/z 319 [M+lf.

Example 25. Synthesis of 5-( { 7-(cyclopropylamino)-5-(4,4-difluoropipcridin- 1 -

[00312] (Z)-5-((5-(Chloro-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3- yl)rnethyiene)imidazolidine~2,4-dione (25 mg, 0.08 mmol) was suspended in NM P (0.2 mL). 4,4-difluoropiperidine hydrochloride (60 mg, 0.38 mmol) and diisopropylethyl amine (67 ,uL, 0.38 mmol) were added and the reaction was irradiated in the microwave at 140 °C for 20 min. The product was purified by preparative HPLC to afford (Z)-5-((7-(cyclopropyiamino)-5-(4,4- difluoropiperidin- 1 -yl)pyrazolo[ 1 ,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (5.9 mg, 14%). LCMS (ES): >95% pure, m/z 404 [M+I ]⁺. Example 26. Synthesis of -(cyclopentylarnino)-7-(cyc

a]pyrimidine-3 -carbaldehyde .

[00313] Terr-butyl 5-chloro-3-formylpyrazolo[l,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (1 .0 eq, 105 mg, 0.312 mmol) was dissolved in DMF (1 ml) in a vial. K₂C0₃ (1.5 eq, 64 mg, 0.463 mmol) and cyclopentylamine (1.5 eq, 46 ul, 0.465 mmol) were added and the mixture was stirred at 50"C for one hour. An additional amount of cyclopentylamine (1.5 eq, 46 ul, 0.465 mmol) was added and the mixture stirred at 70°C for 2 hours. Water was added and the resulting precipitate was filtered and dried to provide 130 mg of solid. This solid was stirred in HC1 4N in dioxane (4 mi) at room temperature for 4 hours. Methanol (1 ml) and aqueous 6N HC1 (2 ml) were added and the mixture stirred at room temperature overnight. The reaction was

subsequently stirred overnight at 60°C to complete the imine hydrolysis. The reaction was neutralized with 6N NaOH and the compound extracted with methylene chloride. After drying over Na₂S0₄, and evaporation of the volatiles, the material was triturated in ethyiacetate to form a solid. 5-(cyclopentylamino)-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde was isolated as a solid by filtration (2!mg). LCMS (ES):>95% pure, m/z 286 i M ! ! Γ .

Example 27. Synthesis of 5-((5-(cyclopentylamino)-7-(cyclopropylamino)pyrazolo[ 1 ,5- alpyrimidin-3- l)methylene)imidazolid.ine-2,4-dione.

[00314] 5-(Cyclopentylarnino)-7-(cyclopropylamino)pyrazolo[l,5-a]pyriniidine-3- carbaldehyde (1.0 eq, 20 mg, 0.070 mmol) was mixed in a vial with hydantoin (2.8 eq, 20 mg, 0.20 mmol) in Ethanol (0.3 ml). Piperidine (2.9 eq, 20 ul, 0.202 mmol) was added and the mixture was stirred at 90°C for 3 hours. The mixture was cooled down, the precipitate was filtered, washed with ethanol and dried. (Z)-5-((5-(cyclopentyiamino)-7- (cyclopropylamino)pyrazolo[l,5-a]pyriiTiidin-3-yl)methylene)imi was isolated as a solid (25 mg, 100%). LCMS (ES):>95% pure, m/z 368 [M+H]⁺.

[00315] The compounds listed in the following Table 6 are Example 25, Example 26, and Example 27.

Table 8.

Example 28. Synthesis of ( ,Z)-5-((7-(cyclopropylamm

yl)pyrazolo[l,5-a]pyrimidin-3-yi)methylene imidazoiidine-2,4-dio

[00316] (R.,Z)-5 ~((7-(Cyclopropylamino)-5 -(3 -fluoropyrroli din- 1 -y3 )pyrazol o [1 ,5- a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione was prepared using chemistries similar to the ones used to prepare (Z)-5-((5-(cyclopentylamino)-7-(cyclopropylamino)pyrazolo[ 1 ,5- a]pyrimidm-3-yi)methylene)imidazolidine-2,4-dione. The compound was isolated as a solid (84 mg, 75% yield). LCMS (ES):>95% pure, m/z 372 [M+H]⁺.

Example 29. Synthesis (Z,)-5-((5-(( lr,4r -4-arninocvclohexylamino -7- (cyclopropylamino)pyrazolo[l,5-afe

2 ,2 ,2-trifluoroac etate .

[00317] S-Chloro-S-foraiylpyrazolotl jS-aJpyrimidin-T-y^cyclopropy carbaraa-te (1 ,0 eq, 1 13 mg, 0,335 mmol) was mixed in a vial with trans-tert-butyl 4-aminocyclohexylcarbamate (1.0 eq, 81 mg, 0,335 mmol) and K₂CO₃ (5.0 eq, 232 mg, 1 ,68 mmol) in DMF (1 ml). The mixture was stirred at 70"C for 2.5 hours. Water was added, the solid was filtered and dried. The compound was treated with hydantoiii (3.0 eq, 100 mg), piperidine (3,0 eq, 100 ui) in ethanol (2 ml) at 85- 90°C for 4.5 hours. Water was added and the solid was filtered and dried. The crude solid was suspended in methylene chloride (5 ml) and trifluoroacetic acid (1 ml) and the mixture was stirred at room temperature for 1 hour. The volatiles were evaporated. The residue was dissolved in methanol and water and subjected to purification by preparative HPLC. After genevac evaporation (Z)-5-((5-((lr,4r)-4-aminocyclohexylamino)-7-(cyclopropylamino)pyrazolo[l ,5- a]pyrimidm-3-yi)methylene)imidazoiidine-2,4-dione 2,2,2-trifluoroacetate was isolated a yellow solid (96 mg, 56% yield), LCMS (ES):>99% pure, m/z 397 | M · 1 1 j , Two isomers detected (ratio: 97.5% and 2.5%).

[00318] The following compounds were prepared using chemistries similar to the one used in Example 26, Example 27, Example 28 and Example 29. Compounds were characterized by LCMS. Table 9B shows the biological activities of the compounds listed in Table 9 A.

Table 9A.

101 CK2: ICSO ΡΪΜ2: ICSO AB: MDAMB4S3 AB: BxPC3

Compounds

(5 μΜ ATP) (μΜ)

W2 <0.0i 0.8771

X2 <0.01 1.952

Example 30. Scheme Synthesis oi^" \-( ( 1 r.4r $-4-(7-( c c 11 i o n \ i an s i ¾ i ·; » )-3-¾ ( H 2,5- dioxoinu^azolidm-4-ylidene)methyl pyrazolo[l,5-a]pyrimidin-5- ylarmno)evclohexyl¼C6tamide

[00319] (Z)-5 -((5 -(( 1 r,4r)-4- Aniinocyclohexylamino)-7-(cyclopropylamino)pyrazolo [1,5- a]pyriiTiidin-3-yl)methylene)imida2;olidine-2,4-dione 2,2,2-trifluoroacetate (1.0 eq, 10 mg, 0.0196 mmol) and DIE A (1.2 eq, 4 ul, 0.0229 mmol) were dissolved in NMP (0.1 mi). Acetic anhydride (1.0 eq, 2 ul, 0.0211 mmol) was added and the mixture stirred at room temperature overnight. Water was added and the resulting precipitate was filtered and dried to provide N- (( 1 r,4r)-4-(7-(cyclopropylamino)-3-((Z)-(2,5 -dioxoimidazolidin-4-ylidene)metiiyl)pyrazolo [1,5- a]pyrimiditi-5-yiamino)cyclohexyl)acetamide as a solid (8 mg). LCMS (ES):>95% pure, m/z 439 | M · l i | .

Example 31. Synthesis of 3 -(( 1 r,4r)-4-(7-(cyclopropylamino)-3 -((Z )-{2,5 -dioxQimidazolidin-4-

trifiuoroacetate

[00320 J (Z)-5-((5-((l.r,4r)-4-Aminocyclohexylamino)- /-(cyclopropylamino)pyrazolo[l ,5- a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione 2,2,2-trifluoroacetate (10 mg) and DIEA (1.2 eq, 4.1 ul) were mixed in dry NMP (0, 1 ml). Dimethylcarbamic chloride (1.0 eq, 1.8 ul) was added and the mixture stirred at room temperature overnight. The reaction was diluted with NMP (1 ,5 ml) and a few drops of water. The compound was purified by preparative HPLC and was isolated after evaporation at the genevac. 3-((lr,4r)-4-(7-(cyclopropyiamino)-3-((Z)-(2,5- dioxoimidazolidin-4-ylidene)niethyl)pyrazoio[ 1 ,5-a]pyrimidin-5-ylamino)cyclohexyi)- 1,1- dimethyiurea 2,2,2-trifluoroacetate. LCMS (ES):>95% pure, m/z 468 [M+H]⁺. Z:E ratio: 86: 13

[00321] The following molecules were prepared using similar chemistries to the ones described in Example 30 and Example 31 using the appropriate amines and anhydrides or acyl chlorides, sulfamoyl chlorides, sulfonyl chlorides or chloroformates. Compounds were purified by preparative HPLC, isolated after genevac evaporation, and characterized by LCMS. Table 10B shows the biological activities of the compounds listed in Table 10A.

Table 10A.

Table 10B. 8 CK2: IC50 PI 2: 1C50 AB: MDAMB453 AB: BxPC3

(μΜ) (5 μ,Μ ATP) (μΜ) (μΜ)

A3 <0.01 > 2.5000 > 30 > 30

B3 <0.1 0.5385

C3 <0.01 0.6791

D3 <0.01 0.476

E3 <0.01 > 2.5000

[00322] The following molecules in Table 10 were prepared using chemistries similar to the ones in Example 26, Example 27, Example 28 and Example 29.

Table 10.

106 Example 32. S nihcsis or /i.77-buiyl 5-(bcnxyhriio)-3-i rn:yipyr;: ^,.>k)[ 1 jp j in:i li: -7- vKcyclopro yQcarbamate

[00323] Diisopropylethylamine (256 μ,Ε, 1.48 mmol) was added to tert-hutyl 5-ch!oro-3- formylpyrazolo[l,5-«]pyrimidin-7-yl(cyclopropyl)carbamate (250 nig, 0.74 mmol) suspended in ethanol (2.5 mL). Benzyl mercaptan (191 uL, 1.48 mmol) was added and the reaction was homogeneous after ~2 min. After 10 min, the reaction was diluted with ethanol (3 mL) and the precipitate was filtered and washed with ethanol (10 mL) to yield tert-hutyl 5-(benzylthio)-3- formylpyrazolo[l,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (148 mg, 47%). LCMS (ES): >95% pure, m/z 425 [M+l ,

Example 33. Synthesis of 5-((5-(benzylt.hio)-7-(cy

3-yr)meth lene)imidazolidme-2,4-dione

[00324] Hydantoin (67 mg, 0.67 mmoi) and piperidine (66 μΐ.,, 0.67 mmol) were added to tert-hutyl 5-( benzylthio)-3-formylpyrazolo[ 1 ,5-a]pyrimidin-7-yl(cyclopropyl)carbamate (95 mg, 0.22 mmol) dissolved in ethanol (1.1 mL). The reaction was heated at 80 °C. After 15 h, the reaction was cooled to r.i, diluted with w^rater (5 mL), and the precipitate was collected and washed with 1 : 1 ethaiiobwater (10 mL) and then ethanol (3 mL). The bright yellow solid was dried in vacuo to give (Z)-5~((5-(benzylihio)-7~(cyclopropylamino)pyrazolo[l ,5- ]pyrimidin-3- yl)methylene)imidazolidine-2,4-dione (75 mg, 66%). LCMS (ES): >95% pure, m/z 507 [M+l]⁺.

[00325] 5-((5-(Berizylthio)-7-(cyclopropylamino)pvra .o3 [l ₅5-a]pvrimidin-3- yl)methylene)imidazolidine-2,4-dione (60 mg, 0.15 mmol) was dissolved in dichloromethane (1 mL) and trilluoroacetic acid (1 mL). After 1 h, the solution was concentrated under a stream of air. The residue was triturated with Et₂0 (3 mL) and the precipitate was collected to provide (Z)-5-((5-(benzylthio)-7-(cyclopropylammo)pyrazolo[l,5-a]pyrirnidin-3- yl)methylene)iniidazolidine-2,4-dione (59 mg, 98%), LCMS (ES): >95% pure, m z 407 [M+l]⁺.

Table 1 1.

Example 34. Synthesis of 5-((5-chloro-7-(cyclopropylammo)pyrazolo[L5-a]pyrimidin-3- yl)methylene)thiazolidine-2,4-dione

[00326] To 5-ehloro-7-(cyciopropylarnino)pyrazoio[ 1 ,5-a]pyrimidine-3-carbaldehyde (440 mg, 1.86 mmol) in EtOH was added thiazolidine-2,4-dione (458 mg, 3.91 mmol) and piperidine (208 μί, 2.05 mmol). The reaction was heated at 80°C overnight. To the reaction mixture, isopropanol (3 mL) was added in the along with 218 mg thiazolidine-2,4-dione and 94μΕ of piperidine. The temperature was increased to 90°C and the reaction was stirred at that temperature overnight. The precipitate was filtered while hot and dissolved in MeOH. To the reaction mixture, 1M HCl (1 mL) was added and the mixture sonicated. The precipitate was filtered and washed with MeOH to yield 340 mg (54% yield) 5-((5-chloro-7- (cyclopropylarnino)pyra.zolo[ 1 ,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione as a yellow powder, LCMS iM \ 336) Example 35. Synthesis of 5-{{5-{3-chlorophenylamino)

a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione

[00327] To 5-((5-chloro-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3- yl)niethyiene)thiazolidine-2,4-dione (20 mg, 0.06 mmol) in NMP was added 3-chloroani ine (38 μΕ, 0.36 mmol) and a few granules of / toluenesulfonie acid. The reaction was heated in microwave at 180°C 1.5 hours. The reaction mixture was filtered and purified by prep HPLC then prep TLX (1 %MeOH/DCM) to yield 5-((5-(3-chiorophenylammo)-7- (cyclopropylamino)pyrazolo[ 1 ,5-a]pyrimidin~3-yi)methylene)thiazoiidiiie-2,4-dione as a yellow solid. I .CMS i M 1 427 )

Example 36.

alpyrimidin--3-yl)methylene)thiazolidine-2,4-dione

[00328] To 5-((5-chloro-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3- yi)methylene)ihiazolidine-2,4-dione (30 mg, 0.09 mmol) in NMP was added 2-methylpropan-l- amine (20 mg, 0.268 mmol). The reaction was heated at 130°C overnight. The reaction mixture was diluted with MeOH and purified by prep HPLC to yield 5-((7-(cyclopropylamino)-5- (isobutylamino)pyrazolo[l

LCMS (M+l=373) [00329] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 36. All compounds were characterized by LCMS. Table 12B shows the biological activities of the compounds listed in Table 12 A.

Table 12A.

Table 12B,

PIM1 : IC50

CK2: IC50 PIM2: JC50 AB: DAMB453 AB: BxPC3

Compound (30 μΜ

(μΜ) (5 μΜ ATP) (μΜ)

ATP)

L3 <1.0 > 2.5000 1.4327

M3 <0.1 1.4455 1.4379

N3 <1 .0 > 2.5000 > 2.5000

03 <0.1 1.2533 > 2.5000

P3 <L0 > 2.5000 > 2.5000

Q3 <0.1 2.0461

R3 <1.0 1.82

Example 37. Synthesis of tert-butyl 5-chloro-3-formylpyrazolof 1 ,5-a]pyrimidin~7~

yl(cyciopropyl)carbamate

[00330] To 5-chloro-7-(cyc[opropylammo)pyrazolo[l,5-a]pyrimidine-3-carbaldehyde (4,52 g, 19,15 mmoi) in methylene chloride (80 mL) was added triethylamine (3.2 mL, 23 mmol), dimethyiammopyridine (350 mg, 2.87 mmol), and di-t-butyldicarbonate (12.53 g, 57,44 mmol) The mixture was stirred at room temperature for 60 minutes, The reaction mixture was transferred to a separatory funnel, washed IX with H₂0, 2X with brine, dried over MgS0₄, filtered, and evaporated to dryness to provide an oily residue. The crude material was purified by silica gel chromatography (0%-20% ethyl acetate/hexanes) to yield 5.68 g (88% yield) of tert-butyl 5-chloro-3-formylpyrazolo[ 1 ,5-a]pyrimidin-7-yi (cyclopropyl)carbamate. LCMS

(M+l = 337) Examp

[00331] To 5 tert-butyl 5-chloro-3-formylpyrazolo[l ,5-a]pyrimidin-7-yl

(cyclopropyl)carbamate (650 mg, 1.93 mmol) in 14 mL of a 2: 1 mixture of 1 ,2- dimethoxy ethane/ EtOH was added 3-hydroxyphenyl boronic acid (399 mg, 2.89 nimol), tetrakis(tripiienylpiiosphine)palladium(0) (1 12 nig, 0.096 mmol), and 2M aqueous solution of ^'Na₂C0₃ (2.9 mL, 5.79 mmol). The mixture was stirred at 85°C for 1 h. The volatiles were removed by rotary evaporation and the residue was purified by silica gel chromatography (0%- 30% EtOAc/Hexanes) to provide 400 mg of tert-butyl cyclopropyl(3-formyl-5-(3- hydroxyphenyl)pyrazolo[ 1 ,5-a]pyrimidin-7-yl)carbamate. (52%). LCMS (M+l=395 )

Example 39. Synthesis of 7-( cyclopropylamino)-5-(3-hydroxyphenyl)pyrazolo[ 1 ,5- a]pyrimidine-3-carbaldehyde

00332] To tert-butyl cyclopropyi(3~formy4~5-(3-hydroxy henyl)pyrazolo[ 1 ,5-a]pyrimidin-7~ yl)carbamate (400 mg, 1.01 mmol) in methylene chloride (20 mL) was added TFA (10 mL). The reaction mixture was stirred at room temperature for 2 hours. The volatiles were removec by rotar evaporation and the residue was purified by silica gel chromatography (0%~40% EtOAc/hexanes) to provide 103 mg of 7-(cyclopropylamino)-5-(3hydroxyphenyl)pyrazolo [1 , a]pyrimidine-3-carbaldehyde. (35%). LCMS i^'M 1 295 ) Example 40. Synthesis of 5-{{7-{cyclopropylaimno)

a]pyrimidin-3- l)methylene)imidazolidine-2,4-dione

[00333] To 7-(cyclopropylamino)-5-(3hydroxyphenyl)pyrazoio [ 1 ,5-a]pyrimidine-3- carhaldehyde (100 mg, 0.340 mmol) in EtOH (2 ml.) was added piperdine (67 μΕ, 0.680 mmol), and hydantoin (34 mg, 0.34 mmol). The reaction was stirred at 50°C overnight. The solid formed was isolated by filtration to provide 70 mg of 5-((7-(cyclopropylamino)-5-(3- hydroxyphenyl)pyrazolo[l .,5-a]pyTimidin-3-y3)rafithylene)imidazo3.idine-2,4-dione. (55%). LCMS (M+l=377)

[00334] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 38, Example 39, and Example 40. All compounds were characterized by LCMS. Table 13B shows the biological activities of the compounds listed in Table 13A.

Table 13 A.

Table 13B.

CK2: IC50 PI 2: 1C50 AB: MDAMB453 AB: BxPC3

Compound

(μΜ) (5 μ,Μ ATP) (μΜ)

G4 <0.01 > 2.5000 1.153 1.191

H4 <0.01 > 2.5000 1.964 14.486

14 <0.1 > 2.5000 0.683 1.898

J4 <0.01 0.867 4.746 > 30

K4 <0.1 1.3082 1 .938 2.578

L4 <0.01 1.4748 1 .79 0.725

M4 <0.01 1.2497 > 30 14.437

N4 <0.01 > 2.5000 > 30 12,535

04 <0.01 > 2.5000 17.123 1 ^32

P4 <0.01 0.0754 5.276 0.549

Q4 <0.01 0.2562 1.068 0.745

R4 <0.01 0.0487 14.882 10.61

S4 <0.01 > 2.5000 20.012 4.608

T4 <0.1 > 2.5000 1.706 2.744

U4 <0.01 > 2.5000 1.263 8.129

V4 <0.1 > 2.5000 12.417 > 30

W4 <0.01 2.084 12.278 > 30

X4 <0.01 1.7271 > 30 > 30

Y4 <0.01 > 2.5000 1.979 2.253

Z4 <0.01 > 2.5000 15.69 29.035

A5 <0.01 0.948 1.742

B5 <0.01 > 2.5000 26.74 5.426

C5 <1.0 > 2.5000

Synthsis of 5 -(7-( tert-butoxycarbonyl(cyclopropyr)amino)-3 -fomiylpyrazolo [ 1 ,5 -a]pyrimidin-5 - yi)-2~fl uot benzoic acid

[00335] Same procedure as [Example 38] , LCMS ( M · ! 441 ) Example 41. Synthesis of 5-(7-{cyclopropyl3mino)-3-f rrri.ylpyrazoio[l ,5~a]pyrimidm-5-yl)-2~ fluorobenzoic acid

[00336] Same procedure as [Example 39] . LCMS (M+l=341)

Example 42. Synthesis of 7-(cyclopropyiamino)-5-(4-fluoro-3-{moj^*pholine-4- car bonyl)phenyl)pyrazolo [ 1 ,5 -a]pyrimidine-3 -carbaldehyde

[00337] To 5-(7-(cyclopropylamino)-3-fomiylpyrazolo[l,5-a]pyrimidin-5-yl)-2- fluorobenzoic acid (75 mg, 0.22 mmol 0 in DMF (3 niL) was added EDCI (46 mg, 0.24 mmol), HOBt (33 mg, 0.24 mmol), and morpholine (21 mg, 0,24 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with ethyl acetate and washed IX with saturated sodium bicarbonate, 2X with brine, dried over MgS0₄, filtered and evaporated to dryness to provide 92 mg of 7-(cyclopropylamino)-5-(4-fluoro-3-(morpholine-4- carbonyl)phenyl)pyrazoio[l,5-a]pyrimidine-3-carbaidehyde, LCMS (M+l=410)

Example 43. Synthesis of 5 -((7-(cyclopropy laniino)- 5 -(4-fluoro-3 -(morpholine-4 -

00338J Same procedure as [Example 40] . LCMS ( M 1 49 ) [00339] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 42 and Example 43. All compounds were characterized by LCMS. Table 14B shows the biological activities of the compounds listed in Table 14A.

Table 14 A.

Table 14B.

Example 44. Synthesis of tert-butyl cyclopropyl(5-(2-fluoropyridin-4-yl)-3- forrny lpyrazolo[ 1 ,5-a]pyrimidin-7-yl)carbamate

[00340] To tert-butyl 5~chloro~3-formylpyrazoio[ 1 ,5-a]pyriniidin-7~yl(cyclopropyl)carbamaie (1 g, 3 nunol) in 29 mL of a 2: 1 mixture of 1,2-dimethoxyethane/ EtOH was added 2- Fluoropyridine-4-boronic acid (500 mg, 3.55 mmol), telTakis(triphenylphosphine)palladium(0) (173 mg, 0.15 mmol), and 2M aqueous solution of a₂C0₃ (4.4 mL, 8.9 mmol). The mixture was stirred at 85°C for 8 hours. The volatiles were removed by rotary evaporation and the residue was purified by silica gel chromatography (35% EtOAc Hexanes) to provide 324 mg tert-butyl cyclopropyl(5-(2-fluoropyridm-4-yl)-3-fo^

yl)carbamate (28% yield). LC S (M+l=398)

Example 45. Synthesis of 7-(cyclopropy 1 ammo)-5 -( 2-fluoropyridm-4- l)pyrazolo [ 1.5- alpyrimidine-3-carbaldehyde

[00341] To tert-butyl cyclopropyl(5-(2-fiuoropyridin-4-yl)-3-formy[pyrazolo[l,5- a]pyrimidin-7-yl)carbamate (320 mg, 0.82 mmol) in methylene chloride (3 mL) was added TFA. (3 mL). The reaction mixture was stirred at room temperature for 1.5 hours. The volatiles were removed by rotary evaporation and IN NaOH was added to the residue to make basic. The precipitate was collected by filtration, washed with water, and dried under vacuum to provide 180 mg of 7-(cyclopropylamino)-5-(2-fluoropyridin-4-yl)pyrazolo[l ,5-a]pyrimidine-3- carbaldehyde (74%). LCMS (M · 1 298)

Example 46. Synthesis of 5 -((7-(cyclopropylamin o )- -(2-fluoropyri din-4-yl jpyrazo 1 o [ 1 , - alpyrimidin-3- l)methylene)thiazolidine-2,4-dione

[00342] To 7-(cyclopropy 1 amino)~5 ~(2- f! uoropyridin-4-yl)pyrazolo [ 1 ,5 -a]pyrimidine-3 - carbaldehyde 30 mg, 0.1 mmol) in EtOH (1 niL) was added piperdine (13 ί·, 0.1 nimol), and thiazoIidine-2,4-dione (12 mg, 0.1 nimol). The reaction was stirred at 80°C for 2 hours. The solid formed was isolated by filtration, washed with water then ethanol. The recovered solid was further purified by washing with 20% methanol/dichioromethan to provide 9 mg of 5-((7- (cyclopropylamino)-5-(2-fluoropyridin-4-yl)pyrazolo[l ,5-ajpyrira.idm-3- yl)methylene)thiazolidine-2,4-dione (23%). LCMS { M · 1 397)

[00343] The compoimds described in the following table were prepared using chemistries similar to those exemplified in Example 44, Example 45, and Example 46. All compounds were characterized by LCMS. Table 15B shows the biological activities of the compoimds listed in Table 15 A.

Table 15 A.

Table 15B,

Example 47. Synthesis of 5 -((7-(cyc lopropylamin o )-5 -(2 ~fl uoropyr i din-4-yl jpyrazo 1 o [ 1 , - alpyrimidin-3-yl)methylene)-2-thioxothiazolidin-4-one

[00344] To 7-(cyclopropylamino)-5-(2-fluoropyridin-4-yl)pyTazolo[ 1 ,5-a]pyrimidine-3- earhaldehyde 30 nig, 0.1 mmol) in EtOH (1 mL) was added piperdine (13 μΕ, 0.1 mmol), and rhodanine (13 mg, 0.1 mmol). The reaction was stirred at 80°C for 2 hours. The solid formed was isolated by filtration, washed with water then ethanol. The recovered solid was further purified by washing with 20% methanol/dichioromethan to provide 15 mg of 5-({7- (cyclopropylammo)-5-(2-fluoropyri^

thioxothiazolidin-4-one (35%). LCMS ( M · 1 1 )

Table 16.

Example 48.

ylamino ^")piperidine- 1 -carboxylate

[00345] To the reaction flask, 5,7-dichloropyrazolo[l ,5-a]pyrimidine (896 mg, 4.8 mmo was added along with ter -butyl 4-aminopiperidine- 1 -carboxylate (954 mg, 4.8 mmol), triethylamine (664 μΐ,, 4,8 mmol), and aceionitrile (16 ml.). The reaction was heated at 1 00°C for 12 hours then cooled to room temperature, diluted with water, filtered and washed with water. The product, tert-butyl 4-(5-c oropyrazolo[l,5-a]pyrimidin-7-ylamino)piperidme-] - carboxylate, was collected as a solid in quantitative yield and dried under vacuum overnight. LCMS (M+l= 352)

Example 49, Synthesis of tert-butyl 4-(5-chloro-3-formylpyrazolo[ 1 ,5~alpyrimidm-7~

yiamino piperidine- 1 -carbox late

[00346] To tert-butyl 4-(5 -chloropyrazolo [ 1 ,5 -a]pyrimidin-7-ylamino)piperidine- 1 - carboxylate (1.7 g, 4.8 mmol) in DMF (36 mL), POCl₃ (7.7 niL, 82.9 mmol) was added dropwise at room temperature. After the addition was complete, the reaction was stirred for 8 hours. Then, the reaction was quenched by slow addition to ice cold 6N NaOH. The mixture was diluted with water and the solid was collected by filtration. The solid was washed several more times with water then dried under vacuum overnight. The product, tert-butyl 4-(5-chloro- 3-formylpyrazolo[l ,5-aJpyrimidm-7-ylamino)piperidine- 1 -carboxylate, was collected as a solid in 48% yield. LCMS ( M · 1 380)

Example 50. Synthesis of tert-butyl 4-(5-(3-chlorophenylamino)-3-formylpyrazolo l ,5-

[00347] Tert-butyl 4-(5-chloro-3-fom ylpyrazolo[l ,5-a]pyrimidin-7-ylaniino)piperidine-l- carboxyiate (876 mg, 2,3 mmol) was added to 1 ,4-dioxane (6 mL) along with 3-chloroaniline (1.5 mL, 13.9 mmol) and p-toluenesulfonie acid monohydrate (44 mg, 0.23 mmol). The reaction was heated at 95°C for 12 hours then cooled to room temperature, diluted with water, and filtered. The solid w^ras washed with IN NaOH followed with water then dried under vacuum overnight. The product, tert-butyl 4~(5-(3-chloropheriylammo)-3-formy]pyrazolo[i ,5- a]pyrimidin-7-y[amino)piperidine-l-carboxylate, was collected after further purification by recrystallization from ethyl acetate/hexanes (74% yield). LCMS (M+ 1=471)

Example 51. Synthesis tert-batyl 4-(5-(3-chloro

ylidene)methyl)pyrazolo[l ,5-a]pyrimidin-7-ylarrnno)piperidine-l -c

8 J To the reaction flask, tert-butyl 4-(5-(3-chSorophenylamino)-3-formy!pyrazoSo[l ,5- a]pyrimidiii-7-yiamino)piperidine-l-carboxylate (81 1 mg, 1.7 rnmoi) was added to ethanol (6.3 niL) along with hydantoin (172 mg, 1.7 rnmoi) and piperidine ( 170 μΐ,, 1 .7 mmol). The reaction was heated at 80°C for 12 hours then cooied to room temperature and diluted with water. The solid was collected by filtration, washed with water and cold ethanol. The material was dried under vacuum overnight. The product, tert-butyl 4-(5-(3-chiorophenylaniino)-3-((2,5- dioxoirnidazolidin-4-ylidene)methyl)pyrazolo[i ,5-a]pyr^

carboxylate, was recovered as a red solid in 67% yield after further purification by

recrystallization from ethyl acetate/hexanes. LCMS (M+l = 553)

Example 52. Synthesis 5-((5-(3-chlorophenyla.niino)-7-(piperidin-4-yiamino)p\Tazoloi^"L5- a] pyrimidm-3 -ypmethy 1 ene)imidazolidine-2 ,4-di one

[00349] Tert-butyl 4-(5-(3-chlorophenyiarnino)-3-((2,5-dioxoimidazolidin-4~

ylidene)methyl)pyrazolo[ 1 ,5-a]pyrimidin-7-ylamino)piperidine- 1 -carboxylate (640 mg, 1 ,2 mmoi) was dissolved in 10 ml of TFA/DCM (1 : 1 ) and stirred at room temperature for 1 hour then quenched by addition to ice cold 6N NaOH. The mixture was diluted with water then the aqueous layer was decanted. The organic layer was diluted with hexanes and iiltered. The product, 5-((5-(3-chlorophenylamino)-7-(piperidin-4-ylamino)pyrazo!o[l ,5

yl)methylene)imidazolidine-2,4-dioiie, was collected as a solid in quantitative yield. LCMS ( M I = 453)

Example 53. Synthesis of 5-((5-(3-chlorophenyiamino -7-(l -(cycjopropanecarbonyj )piperid

4-ylamiiio) razolo[l ,5-a]pyrimidin-3-yl)methyk¾ie)imidazolidine-2,4-

[00350] To 5-((5-(3-chiorophenylamino)-7-(piperidin-4--yiamino} pyrazoio [1 , 5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4~dione (30 mg, 0.066 mmoi) in THF was added cyclopropyl carbonyl chloride (5 iL, 0.04 mmoi). The mixture was stirred at room temperature for ten minutes. The reaction mixture was then concentrated, diluted with MeOH, and purified by prep HPLC to yield 5-((5-(3-chlorophenylamiiio)-7-(l-(cyciopropanecarbonyi)piperidiii-4~ ylamino)pyra olo[l ,5-a]pyrimidm-3-yl)methylene)imidazolidine-2,4-dione. LCMS (M+l ^::::521 )

Example 54.

yIamjno) razolo[l,5-a]pyrimidin-3-yl)methylene)imi

51 J Same procedure as [Example 53]. LCMS (M+l=53v) Example 55, Synthesis of 5-((5-(3-chlorophenylamino)-7-(l-(3,3-diniethylbutanoylpiperidm- 4-ylamino) yrazolo[ L5-a]pyrjmidin-3-yl)methvjene)imidazolidme-2,4-diorie

[00352] Same procedure as [Example 53]. LCMS (IV! I 551}

Example 56. Synthesis of 4-{5-{3-chlorophenylammo)

ylidene)methyl)pyrazolo[l,5-a]pyrimidin-7-y^

carboxamide

[00353] Same procedure as [Example 53] . LCMS (M+ 1=524)

Example 57. Synthesis of methyl 4 -{5 -(3 -chlorophenylamino )-3 -((2,5 -dioxoimidazolidin-4- ylidene)meth l)pyrazolo[1.5-a3pyrimidin-7-ylaniino)piperi

[00354] Same procedure as [Example 53] except DMF is used as solvent,

LCMS (M+l=511)

Example 58. Synthesis of methyl 2-{4-(5-(3-ehloropheny

ylidene)methyl)pyrazolo[ 1 ,5-a]pyrimidin-7-ylammo )piperidin- 1 -yl)acetate

[00355] Same procedure as [Example 53] except DMF is used as solvent.

LCMS (M ! 525 ;

Example 59. S nihcsis or 5 i5-C3~chiorop enyiamii :o s-7~( i -(2-hyd X)xypropyi ;plpcriiiiii-4- yIamjno) razolo[l,5-a]pyrimidin-3-yl)methylene)imi

[00356] To 5-((5-(3-chlorophenyla,mino)-7-(piperidin-4-ylamino)pyrazolo[ 1 ,5-a]pyrimidin-3- yl)methy!ene)imidazolidme-2,4-dione (30 mg, 0.066 mmol) in DMF was added 1 -chloro2- propa.no! (7 μΐ,, 0.13 mmol) and potassium iodide (11.0 mg,0.066 mmol). The mixture was heated to 120°C and stirred for overnight. The reaction mixture was concentrated , diluted with MeOH, and purified by prep HPLC to yield 5~((5-(3-chlorophenyiamino)-7-(1 -(2- hydroxypropyl)piperidin-4-ylamm^

2,4-dione. LCMS (M+ 1=511) Example 60.

yIamjno)pyrazolo[l,5-a]pyrimidin-3-yl)methylene)im

[00357J Same procedure as [Example 59] . LCMS (M+l.=497) Example 61. Synthesis or ^-l l S-C !- ii soi i henviai iino i- ?-! I pynuin-2-vinie hyi )pipcndin-4- ylamino) razolof 1 ,5-alpyrimidin-3-yl)mefeylene)imidazolidine-2,4-dione

[00358] To 5-((5-(3-chlorophenylammo)-7-(piperidin-4-ylamino) pyrazolo [1,5-a] pyrimidin- 3-yi) methylene) imidazoiidine-2,4-dione (30 mg, 0.066 mmol) in DMF was added 2- (bromomethyi) pyridine hydrogen bromide (26.0 mg, 0103 mmoi), The mixture was stirred at room temperature for 0.5 hour. The reaction mixture was concentrated , diluted with MeOH, and purified by prep HPLC to yield 5-((5-(3-chlorophenylamino)-7-(l-(p>Tidm-2- ylmethyi)piperidin-4-ylamino)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4- dione. LCMS (M+ 1=544) Example 62.

yIamjno) razolo[l,5-a]pyrimidin-3-yl)methylene)imi

[00359] To 5-((5-(3-chk>rophenylamino)-7-(piperidin-4-ylamino) pyrazolo [1, 5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4-dione (20 mg, 0.04 mrnoi) in THF and AcOH (4.8 mg, 0.08 mmol) was added acetone (2.0 mL, 0.2 mmol) and sodium triacetoxy borohydride (85.0 mg, 0.4 mmol). The mixture was heated at 60°C for one hour. Saturated sodium bicarbonate solution was added to the reaction mixture. The mixture was extracted with ethyl acetate and dried over sodium sulfate. Then the mixture was , diluted with MeOH, and purified by prep HPLC to yield 5-((5-(3-chlorophenylamino)-7-(l-isopropylpiperidin-4-ylamino) pyrazoio [1, 5-a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione. LCM8 ( + 1=495)

Example 63. Synthesis of 5-((5-(3-chlorophenylamino)-7-(l-ethvipiperidin-4- ylamino) razolo l,5-a]pyrimidin-3-yl)methylene)imidazoli

[00360] To 5-((5-(3-chiorophenylaniino)-7-(piperidiii-4--yiamino) pyrazoio [1, 5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4-dione (30 mg, 0.06 mmol) in THF and AcOH (4.8 mg, 0.08 mmol) was added acetaldehyde (2.0 mL, 0.2 mmol) and sodium triacetoxy

borohydride (85.0 mg, 0.4 mmol). The mixture was stirred at room temperature for 0.5 hour. The reaction mixture was concentrated , diluted with MeOH, and purified by prep HPLC to yield 5-((5-(3-chloropheny3.amino)-7-(l-ethylpiperidm-4-ylamino) pyrazolo [1 , 5~a] pyrimidin- 3-yl) methylene) imidazolidine-2, 4-dione, LCMS (M+ 1=481)

Example 64. Synthesis of 5-((5-( 3-chlorophenylammo)-7-( 1 -isobutyrpiperidin-4- yiamimu rg/oio[ ; .5 jpys insiciin- -yj ¾incth yic $irs ida oiidinc-2.4-dioi o

[00361] To 5-((5-(3-chlorophenylamino)-7-(piperidin-4-ylamino) pyrazolo [1 , 5-a] pyrimidin-3-yl) methylene) imidazolidme-2,4-dione (30 mg, 0.06 mmol) in THF and AcOH (4.8 mg, 0.08 mmol) was added isobutryldehyde (2.2 mL, 0.2 mmol) and sodium triacetoxy borohydride (85.0 mg, 0.4 mmol). The mixture was stirred at room temperature for 0.5 hour. The mixture was concentrated, diluted with MeOH, and purified by prep HPLC to yield 5-((5- (3-chlorophenylamino)-7-(l-isobuty[piperidin-4-ylamino) pyrazolo [1 , 5-a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione. LCMS (M I 509)

[00362] The compounds described in the following table were prepared using chemistries similar to those exemplified in the Examples described above. All compounds were

characterized by LCMS. Table 17B shows the biological activities of the compounds listed in

Table 17A.

Table 17A.

Table 17B.

Example 65. Synthesis of 7-(benz lthio)-5-chloropyrazolo[l ,5-a]pyrimidine

[00363] To the reaction flask, 5,7-dichloiOpyrazolo[l,5-a]pyrimidine (4.1 g, 22 mmol) was added along with benzyl mercaptan (2.8 mL, 22 mmol), triethylamine (3.1 mL, 22 mmo!), and acetonitrile (71 mL). The reaction was stirred at room temperature for 3 hours then diluted with water, filtered and washed with water. The product, 7-(benzy ithio)-5-chloropyrazolo[ 1 ,5- ajpyrimidine, was collected as a solid in 96% yield after drying under vacuum overnight.

I . CMS ( · i 276)

Example 66.

[00364] To the reaction flask, 7-(benzylthio)-5-chloropyrazolo[l,5-a]pyrimidine (3,45 g, 12,5 mmoi) was added along with 3-chloroaniline (3,3 niL, 31.3 mmol), 4N HC1 in dioxane (3,1 mL, 12,5 mmol), and ethanol (42 mL). The reaction was stirred at reflux for 12 hours then cool to room temperature. Excess solvent was removed under vacuum and the residue was diluted with water. The mixture was made basic with 3N NaOH, filtered and washed with water. The product, 7-(benzylthio)-N-(3-chiorophenyl )pyrazolo[l ,5-a]pyrimidin-5-amine, was collected as a solid in 90% yield after drying under vacuum overnight. LCMS (M+l= 376)

Example 67. Synthesis of 7-(benzylthio)-5-(3-chlorophenylamino)pyrazolo[ 1 ,5-a]pyrimidine- 3-carbaldeh de

[00365] To 7-(beazylthio)-N-(3-chlorophenyl)pyrazolo[ 1 ,5-a]pyrimidin-5-amine (4.1 g, 11 ,3 mmoi) in DMF (42 mL), POCl (6.3 mL, 67.6 mmol) was added dropwise at room

temperature. After the addition was complete, the reaction was stirred for 3 hours at room temperature. Then, the reaction was quenched by slow addition to ice cold 6N NaOH. The mixture was diluted with water and the solid was collected by filtration. The solid was washed several more times with water then dried under vacuum overnight. The product, 7-(benzylthio)~ 5-(3-chlorophenyla.mino)pyra.zolo[l,5-a]pyrimidine-3-carbaldehyde, was collected as a solid in 83% yield. LCMS (M · 1 395)

Example 68. Synthesis 5 -((7-(benzylthio)- 5 -(3 -chloropheny I amino) razolo [ 1 ,5 -ajpyrimi din- 3- l)methylene)imidazolidine-2,4-dione

[00366] To the reaction flask, 7~(benzyithio)-5-(3~chiorophenylamino)pyrazolo[l ,5- a]pyrimidine-3-carbaldehyde (3,7 g, 9.3 mmol) was added to ethanol (31 mL) along with hydantoin (933 mg, 9,3 mmol) and piperidine (920 p.L, 9.3 mmol). The reaction was heated at 80°C for 3 days then cooled to room temperature and diluted with water. The solid was collected by filtration, washed with water, 50% ethanol/ water, and then 100% ethanol. The material was dried under vacuum overnight. The product, 5-((7-(benzy!thio)-5-(3- chlorophenyla.mino)pyra.zolo[l,5-a]pyrimidin-3-yl)methylene)imidazolidine-2,4- was recovered as a yellow solid in 92% yield. LCMS (M+l = 477)

Example 69. Synthesis of 5-((7-(berjzyisultlnyl)-5-(3-chlorophenylamino)pyrazolo[l ,5- a rimidin-3 -yl)methylene)imidazolidine-2 ,4 -dione

[00367] To the reaction flask, 5-((7-(benzy[thio)-5-(3-ch[orophenylamino)pyrazolo[l,5- a]pyrimidin-3-yl)methylene)imidazolidiiie-2,4-dione (4.1 g, 8.6 mmol) was added to dichloromethane (86 mL) along with m-chloroperbenzoic acid (5.9g, 34.4 mmol). The mixture was allowed to stir at room temperature for 12 hours. The solid was collected by filtration, washed with dichloromethane then dried under vacuum overnight. The product, 5-((7- (benzy Isulfmy l)-5 -(3 -cMorophenylamino)pyrazolo [ 1 ,5 -a.]pyrimidin-3 - yl)rneihylene)imidazolidine~2,4-dione, was recovered as a bright yellow solid in quantitative yield. LCMS (M+l = 493)

[00368] The compounds described in the following table were prepared using chemistries similar to those exemplified in the Examples described above. Ail compounds were

characterized by LCMS, Table 18B shows the biological activities of the compounds listed in Table 18 A.

Table 18 A.

Table 18B,

Example 70. S nihcsis i ^' 5-((5-( -chioi phcn !ai^ i i H i i-7-( 2-l:y( ¾i xycihyi

amino) razolo[ 1 ,5-alpyrimidin-3-yl)methylene^

To 5-((7-(¾enzylsulfmyl)-5-(3-chlorophenylam.ino) pyrazolo [1 ,5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4-dione (15 mg, 0.0304 mmol) i NMP was added 2-aminoethanol (14.6 uL, 0.242 mmol). The mixture was heated in the microwave at 120 °C for 20 minutes. Water was added to the reaction mixture and the precipitate was collected by filtration. The precipitate was washed with methanol to yield 5-((5-(3-chlorophenylamino)-7-(2- hydroxyethylamino) pyrazolo [1 , 5-a] pyrimidin-3-yl) methylene) imidazolidine-2,4-dione LCMS (M+l=414). Similar products (shown below^r) were also obtained as precipitates by addition of water whil e other reactions were purified by prep HPLC to y ield corresponding products.

Example 71. Synthesis of 5 -((5 -(3 -ehloropheny larriino)-7-f yridin-3 -

Same procedure as [Example 70] . LCMS (M+ 1=461) Example 72. Syni csis 3-{{5-(3-i:hlor )p ci yjan¾l;u ;-7-(pyriiiiii-4- ylmethylamino)pyrazolo[l,5-a]pyrimidin-3-yl)methylene)imid

00371] Same procedure as [Example 70] . LCMS (M+ 1=461)

Example 73, Synthesis 5-((5-(3-chlorophenylamino)-7-(2-(d.imethylamiiio)ethyl amino ^") razo I o [ 1 ,5 -a] pyr imidin-3 ~yl)methy lene)imi dazolidine-2 ,4-di one

[00372] Same procedure as [Example 70] . LCMS ( hi 1 441

Example V4. Synthesis of 5-((5-(3-chlorophenylamino

al yrimidin-3 -yl)methy lene)imida.zolidme-2 ,4 -dione

[00373] Same procedure as [Example 70], LCMS (M+l=412)

Example 75,

ropyIamjno)pyrazolo[l,5-alpyrimidin-3-yl)niethylene)imi^

[00374] Same procedure as [Example 70]. LCMS (M+ 1=428)

Example 76. Synthesis of 5-((5-(3-chlorophenylaniino)-7-(cyclobutyl amino )pyrazolo[^~ 1 ,5- ai vrini jin-3-yi )niclhvienc)n)d ia/oiidinc-2.4-iiic)ii

00375] Same procedure as [Example 70]. LCMS ( M 1 42 )

Example 77. Synthesis of 5-{{5-{3-chlorophenylamino)

amino ) razolo l,5-alpyrimidin-3-yl methylene)iniidazolidine-2,4-dione

00376] Same procedure as [Example 70] . LCMS (M+

Example 78. Synthesis 5-((5-(3-c ^

amino )pyrazoio l,5-alpyrimidin-3-yl)methylene)iniidazolidine-2,4-dione

00377] Same procedure as [Example 70] . LCMS (M+ 1=461)

Example 79. Synthesis of 5-(^

dimethylpropylaminopyrazolo[l,5-a]pyrinn^in-3-yl)meth lene)imidazolidine-2,4-dion^

J Same procedure as [Example 70]. LCMS (M+ 1=483)

Example 80. Synthesis tert-butyl 2-((5-(3-chlorophenyiamino)-3-((^'2,5-dioxoimidazolidrn-4- ylidene)methyl)pTazolori,5-a]pyrimidin-7-ylamino)methyl) yrroli

)379] Same procedure as [Example 70]. LCMS (hi · ! 553)

Example 81.

ylidene)methyl)pyrazolo[ 1 ,5-a]pyrimidin-7-ylamino )niethyl)piperidine- 1 -carboxyiate

[00380] Same procedure as [Example 70] . LCMS (M+l=568) Example 82. Synthesis of 5 -((5 -(3 -chloroph eny lamino)- 7~i 2,2 , 2-tri fl uoroeth yl

amino )pyrazolo l,5-alpyrimidin-3-yl methylene)iniidazolidine-2,4-dione

00381] Same procedure as [Example 70] . LCMS (M+ 1=452)

Example 83. Synthesis of 5 -(( ?-{ 1 H-pyrazo 1 -3 -ylamino)~5 -(3 -chl orophenyl

amino ) razolo l,5-alpyrimidin-3-yl methylene)iniidazolidine-2,4-dione

■ame procedure as [Example 70] . LCMS (M+ 1=436) Example 84. Synthesis 5-((5-(3-chlorophenylamino)-7-(pyrrolidin-3-yl amino)pyrazolo[l ,5- •ijpyi'lsiiiciin-}-)'! jnicihyicncjiinida/ojidii c- ^-dioi c

[00383] Same procedure as Example 70. LCMS ( M · ! )

[00384] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 70. All compounds were characterized by LCMS. Table 19B shows the biological activities of the compounds listed in Table 19 A.

Table 19A.

Table 19B.

CK2: IC50 ΡΙ 2: 1C50 ΑΒ: MDAMB453 ΑΒ: BxPC3

(μΜ) (5 μΜ ΑΤΡ) (μΜ) (μΜ)

Q6 <1.0 1 ,2876

R6 <1.0 1.1213

S6 <0.1

Τ6 <0.1

U6 <1 .0 0.5149

V6 <0.1

W6 <1 .0

Χ6 <1.0

Υ6 <0.1

Ζό <0.1

Α7 <0.1

Example 85. Synthesis of tert-but l 3-(5-cMoropyrazolo[l,5-ajpyrimidin-7-ylamino)benzoate

[00385 J To the reaction flask, 5,7-diehioropyrazoio[! ,5-a]pyrimidine ( 1.6 g, 8.2 mmol) was added along with tert-butyl 3-aminobenzoate (1.7 g, 8.7 mmol), triethylamine (1.2 mL, 8.6 mmol), and t-butyl alcohol (22 mL). The reaction was heated at ! 0Q°C for 6 hours then diluted with water, filtered and washed with water. The product, tert-butyl 3-(5-chloropyrazolo[ 1 ,5- a]pyrimidin-7-yiamino)benzoate, was collected as a solid in quantitative yield after drying under vacuum overnight. LCMS (M+l= 345 }

Example 86. Synthesis of tert-butyl 3 -(5 -(3 -chloropheny lamino)pyrazolo 1 ,5 -alpyrimidin-7- vlamino)benzoate

[00386] To the reaction flask, , tert-butyl 3-(5-chloropyrazolo[l ,5-a]pyrirnidin~7- ylamino)benzoate (2.9 g, 8,2 mrnol) was added along with 3-chloroaniline (2.2 mL, 20.6 mniol), 4N HC1 in dioxane (2.6 mL, 10.4 mrnol), and t-butyl alcohol (41 mL). The reaction was stirred at 100°C for 2 days then cooled to room temperature. The mixture was diluted with water, made basic with 3N NaOH, filtered and washed with water. The product, tert-butyl 3-(5-(3- chiorophenylammo)pyrazolo[ 1 ,5-a]pyrimidin-7-ylamino)benzoate, was collected as a solid in 55% yield after drying under vacuum overnight. LCMS (M+l= 436)

Example 87. Synthesis of tert-butyl 3-(5-(3-chlorophenylarrnno)-3-formylpyrazolo[l ,5- a] rimidin-7-ylamino)benzoate

[00387] To tert-butyl 3-(5-(3-chlorophenylamino)pyrazo[o[l,5-a]pyrimidin-7- ylamino)benzoate (965 mg, 2.2 mmol) in DMF (8.2 mL), POCl₃ (1.2 mL, 13.3 mrnol) was added dropwise at room temperature. After the addition was complete, the reaction was stirred for 3 days at room temperature. Then, the reaction was quenched by slow addition to ice cold 6 NaOH. The mixture was diluted with water and the solid w^ras collected by filtration. The solid was washed several more times with water then dried under vacuum overnight. The product, tert-butyl 3-(5-(3-chlorophenylamino)-3-formylpyrazolo[l ,5-a]pyrimidin-7- ylamino)benzoate, was collected as a solid in 12% yield after purification by column

chromatography on silica using 5% acetone/dichloromethane as the eiuent. LCMS (M+l™ 464)

Example 88. Synthesis of te^

lidene)meth l)pyrazolo[^"L5-a]pyrimidm-7-ylamino)benzoate

[00388] To the reaction flask, tert-butyl 3-(5-(3-chlorophenylarnino)-3-forniylpyrazolo[l ,5- a]pyrimidm-7-y[amino)benzoate (122 mg, 0,3 mmol) was added to ethanol (1.3 mL) along with hydantoin (26 mg, 0.3 mmol) and piperidine (26 μΐ., 0.3 mmol). The reaction was heated at 80°C for 2 hours in the microwave then cooled to room temperature and diluted with water. The solid was collected by filtration, washed with water, 50% ethanol/water, and then 100% ethanol. The materia! was dried under vacuum overnight. The product, tert-butyl 3-(5-(3- dilorophenylaiTdno)-3-((2,5-dioxoim^

ylamino)benzoate, was recovered as a solid in 69°/» yield. LCMS (M+l = 546)

Example 89. Synthesis of 3-(5-(3~chiorop

ylidene)meth l)pyrazolo[ 1 ,5-a]pyrimidin-7-ylamino)benzoic acid.

[00389] Tert-butyl 3 -(5 -(3 -chlorophenylamino)-3 -((2, 5 -dioxoimidazolidin-4- ylide«e)raethyl)pyrazolo[1 ,5-a]pyrimidin-7-ylajmino)benzoate (97 mg, 0.2 mmol) was dissolved in 2 mL of TFA/DCM (1 : 1) and stirred at room temperature for 1 hour. Excess solvent and TFA were removed by evaporation under a stream of nitrogen. The residue was diluted with water then the mixture was filtered. The product, 3-(5-(3-chlorophenylamino)-3-((2,5- dioxoimdazolidin-4~ylidene)m acid, was collected as a solid in 85% yield. LCMS (M+l = 490)

Example 90. Synthesis of tert-butyl 4-¾3-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-

[00390] To the reaction flask, 3-(5-(3~chlorophenylamino)-3~((2,5~dioxoimidazoiidin~4~ ylidene)methyl)pyrazoio[l,5-a]pyrimidin-7-ylamino)benzoic acid (30 mg, 0.06 mmol) was added to DMF (0,5 nil .) along wit HOBi (9.2 mg, 0.06 mmol), triethylamine (8,4 μΕ, 0.06 mmol) and tert-butyl piperazine-1 -carboxylate (1 1 .2 mg, 0.06 mmol). The reaction mixture was stirred at room temperature for 5 minutes then EDC (11.5 mg, 0.06 mmol) was added. The reaction was allowed to stir for an additional hour then diluted with water and filtered. The recovered solid was washed with more water followed by ethanol. The product, tert-butyl 4-(3- (5 -( 3 -chl orophenylamino)-3 -((2 ,5 -dioxoimi dazoiidin-4-yli dene)methyi)pyrazo So[l ,5- a]pyrimidin-7-ylamino)benzoyl)piperazine-l -carboxylate, was collected as a solid in 73% yield. I . CMS (M+l - 658)

Example 91. Symhcsis of 5-{(5-(3- i c¾rophcn hinsino>-7-(3-(plpcr;i inc- i - carbonyl)phenylamino)pyrazoto

dione

[00391] Tert-butyl 4-(3-(5-(3-chlorophenylamino)-3-((2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazoio[1 ,5-a]pyrimidin-7-ylamino)benzoyl)piperazine-l -carboxylate (27 mg, 0.04 mmol) was dissolved in 2 mL of TFA/DCM (1 : 1) and stirred at room temperature for 1 hour. Excess sol vent and TFA were removed by evaporation under a stream of nitrogen. The residue was diluted with water then the mixture was filtered. The recovered solid was washed with water followed by 50% ethanol. The product, 5-((5-(3-chlorophenylamino)-7-(3-

(piperazine- 1 -carbonyl }phenylamino)pyrazolo[ 1 ,5-a]pyrimidin-3-yl)methylene)imidazolidine- 2,4-dione, was collected as a solid in 21% yield. LCMS (M+l = 558) Example 92. Synthesis of -((5-(3-chlorophenyjam

dione

[00392] Same procedure as [Example 90] . LCMS (M+l -586)

[00393] The conipoirads described in the following table were prepared using chemistries similar to those exemplified in the Examples described above, All compounds were characterized by LCMS. Table 20B shows the biological activities of the compounds listed in Table 20A.

Table 20A.

Table 20B.

Example 93. Synthesis of 3-((7-chloropyrazolo[ 1 ,5-a]pyrimidin-5-yl)methyl )benzonitrile

To the reaction flask, 5,7-dichloropyrazolo[l ,5-a]pyrimidine (452 mg, 2.4 mmol) was added along with (3-cyanohenzyl)zinc(TI) bromide (6 mL, 3.75 mmol, 0.625M in DMF), Pdi PPb 0· · (110 mg, 0, 1 mmol), and DMF (10 mL). The reaction was heated at 60°C for 4 hours then cooled to room temperature. The reaction mixture was poured into saturated aqueous NH₄C1 solution and ice and extracted with ethyl acetate. The combined extracts were washed with water, saturated NaCl solution, and then dried over NaiSC^. The solvent was removed in vacuo and the residue was purified by column chromatography on silica using 35% ethyl acetate/hexanes as the eluent. The product, 3-((7-chloropyrazolo[l,5~a]pyrimidin-5- yl)methyl)benzonitrile, was recovered in 64% yield. LCMS (M+l= 269) Example 94. S nihcsis of 3 ί 7-f cyclopropyi nnno )pyr /oK>[ 1 .. ajpynrnldin- - yr)methyl)benzonitrile

[00395] To the reaction flask, 3-((7-chloropyrazolo[l ,5-a]pyrimidin-5-yl)methyl)benzonitrile (400 mg, 1 ,5 mmol) was added along with cyclopropyl amine (1 15 μΐ., 1 .6 mrnol), triethylamine (230 μΕ, 1 ,6 mmol), and acetonitrile (3 mL). The reaction was stirred at room temperature for 8 hours at 80°C then cooled to room temperature, diluted with water, filtered and washed with water. The product, 3-((7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidin~5- yl)methyi)benzonitrile, was collected as a solid in 83% yield after drying under vacuum overnight. LCMS ( M · ! 290)

Example 95. Synthesis of 3-((7-(cyclopropylamino)-3-forrnylpyrazolo[ 1 ,5-ajpyrimidin-5- yl)methyl)benzonitrile

[00396] To 3-((7-(cyc[opropylamino)pyrazo[o[ 1 ,5-a]pyrimidin-5-yl)methyl }benzonitrile (69 mg, 0,24 mmol) in DMF (0.6 mL), POCl₃ (1 30 p L, 1.4 mmol) was added at room temperature. After the addition was complete, the reaction was stirred for 1 hour at room temperature. Then, the reaction was quenched by addition to ice cold 6N NaOH. The mixture was diluted with water and the solid was collected by filtration. The solid was washed several more times with water then dried under vacuum overnight, The product, 3-((7-(cyclopropylamino)-3- formylpyrazolo[l ,5-a]pyrimidin-5-yl)methyl)benzonitrile, was collected as a solid in 37% yield. LCMS (M+l= 318) Example 96. S nihcsis of 3-(( T-Cc cjo roj s hi?vjinc> ?-3-({2,5-ds xo;n:i ia/olicli?:-4- ylidene)methyl) yrazolo[l,5-a]pyrimidin-5-yl)methyl)benzonitrile

[00397] To the reaction flask, 3-((7-(cyclopropylamino)-3-fonnylpyrazolo[l ,5-a]pyririiidm-5- yl)methyl)benzonitri]e (28 mg, 0.09 mmol) was added to ethanol (0.5 mL) along with hydantoin (9 mg, 0.09 mmol) and piperidine (9 μΤ, 0.09 mmol). The reaction was heated at 80°C for 30 minutes in the microwave then cooled to room temperature and diluted with water. The solid was collected by filtration, washed with water, 50% ethanol/water, and then 100% ethanol. The material was dried under vacuum overnight. The product, 3-((7-(cyclopropylamino)-3-((2,5- dioxoimidazolidin-4-ySidene)methyl)pyrazolo[ 1 ,5-a]pyrimidin-5-yl)methyl)benzonitriSe, was recovered as a solid in 34% yield. LCMS (M+l = 400)

Table 21.

Example 97. Synthesis of tert-butyl cyclopix>pyl(3-formyl-5-(3 iydroxyphenyr)pyrazolo[l ,5- 3]pyrimidm-7-yi)carbamate

[00398] To tert-butyl 5~chloro~3~formylpyrazolo[l ,5-a]pyrimidm-7-yl

(cyclopropyl)carbamate (650 mg, 1.93 mmol) in 14 niL of a 2: 1 mixture of 1,2- dimetboxy ethane/ EtOH was added 3-hydroxyphertyl boronic acid (399 mg, 2.89 mmol), tetrakis(triphenylphosphine)palladium(0) (1 12 nig, 0.096 mmol), and 2M aqueous solution of Na₂C0₃ (2.9 mL, 5.79 mmol). The mixture was stirred at 85°C for Ih. The volatiles were removed by rotary evaporation and the residue was purified by silica gel chromatography (0%- 30% EtOAc/Hexanes) to provide 400 mg of tert-butyl cyclopropyl(3-formyl-5-(3- hydroxyphenyl)pyrazolo[l.,5-a]pyrimidin-7-yl)carbamate. (52%). LCMS (M+l=395)

Example 98. Synthesis of 7-(cyclopropy 1 aniino)-5 -( 3 -hydroxy heny 1 )pyrazo 1 o [ 1 ,5- alpyrimidine-3-carbaldehyde

[00399] To tert-butyl cyclopropyl(3-formyl-5-(3-hydroxyphenyl)pyrazolo[ 1 ,5-a]pyrimidin-7- yl.)carbamate (400 mg, 1 ,01 mmol) in methylene chloride (20 mL) was added TFA (10 mL). The reaction mixture was stirred at room temperature for 2h. The volatiles were removed by rotary evaporation and the residue was purified by silica gel chromatography (0%-40%

EtO Ac/hex an es) to provide 103 mg of 7-(cyclopropylamino)-5-(3hydroxyphenyl)pyrazolo [1 ,5- a]pyrimidiiie-3-carbaidehyde. (35%). LCMS (M+l=295)

Example 99. Synthesis of 5 -((7-(cyclopropy lamino)-5 -(3 -hydroxyphen l)pyrazolo [1,5- a rimidin-3-yj)methylene)imidazojidine-2.4-dione

[00400] To 7-(cyclopropylamino)-5-(3hydroxyphenyl)pyrazolo [ 1 ,5-a]pyrimidine-3- carbaldehyde (100 mg, 0.34 mmol) in EtOH (2 mL) was added piperdine (67 μΐ., 0.68 mmol and hydantoin (34 mg, 0.34 mmol). The reaction was stirred at 50°C overnight. The solid formed was isolated by filtration to provide 70 mg of 5~((7-(cyelopropylamino)~5-(3- hydroxypheny[)pyrazoio[l,5-a]pyrimidm^ (55%). LCMS (M ! 377;

Example 100. Synthesis of tert-buty cy clopro py 1(3 - forrny 1 -5 - (3 -

(trifluoromethoxy)phenyl)pyrazolo[l,5-a]pyrimidin-7-yl)carbaniate

[00401] Same procedure as [Example 97] . LCMS (M+l=463)

Example 101. Synthesis of 7-(cyclopropyla.mino)-5-(3-(trifluoromethoxy)phenyl)pyrazolo[ 1 ,5- a] pyri m idine-3 - carbal dehy de

[00402] Same procedure as [Example 98] . LCMS (M+l=363)

Example 102. Synthesis of 5-((7-(cyclopropylammo)-5-(3-

( in i ioronicthoxs )p envi ¾pvra/oio! I J i lpyni iiiiin-3-vi Hi: thvlcnc iiniida oi iiiine-2.4- dione

00403] Same procedure as [Example 99] . LCMS (M ! 445) Example 103. S nthesis of tert-buty 1 cy clopropy 1(3 ~ forrnyl -5 -{3 -

(hydroxymethyl)phenyl)pyrazolo L5-a]pyrimidm-7-y )carbamate

[00404] Same procedure as [Example 97] . LCMS (M+l= 409)

Example 104. Synthesis of 7-(cyclopropyla.mino)-5-(3-(hydroxymethyl)phcnyl)pyrazolo[ 1 ,5- a] pyri m idine-3 - carbal dehy de

00405] Same procedure as [Example 98] . LCMS (M+l= 309)

Example 105. Synthesis of 5-((7-(cyclopropylammo)-5-(3-

00406] Same procedure as [Example 99] . LCMS (M ! 391) Example 106. Synthesis of methyl 3-{7-(tert-butoxyc3rbony j {cyclopropyl)a.nimo)-3- formylpyrazolo[ 1 ,5-a]pyrimidin-5-yl)benzoate

] Same procedure as [Example 97] . LCMS (M+l= 437)

Example 107. Synthesis of methyl 3-(7-(cyclopropylamino)-3-formylpyrazolo[ 1 ,5-a]pyrimidin- 5-yl)benzoate

8] Same procedure as [Example 98] . LCMS (M+l= 337)

Example 108. Synthesis of methyl 3-(7-(cyclopropylarnino)-3-((2,5-dioxoirnidazolidin-4- viidcnc iir: lhvl )pvra/oioi i .5 -a lps nmidin-5--vi ibcn/o;iic

00409] Same procedure as [Example 99] . LCMS (M+l Example 109. Synthesis of methyl tert-butyj cycj opropyl(3-i rmyl-5-{3- ¾methylsulfony)phenyl)pyrazolo[L5-aJpyrimidin-7--yl)carbamate

Exaniple 110. Synthesis of 7-{cyclopropylamino)-5-(3-(methylsulfonyl)phenyl)pyrazolo[ 1 ,5- a] pyri m idine-3 - carbal dehy de

[00411] Same procedure as [Example 98]. LCMS (M+l= 357)

Example 111. Synthesis of 5-((7-(cyclopropylamino)-5-{3-

(rncihvis li nvi¾ph nvi ;py;;i/oi j I ^-aipvriniidin-3-yi jiiclhvienc)n):iiaz iiiiin -2.4- dione

00412] Same procedure as [Example 99]. LCMS (M+l= 439) Example 112. Synthesis of tert-buty cy clopropy 1(3 -formyl -5 - (3 -(N- niethylsulfamoyl)phenyl)pyrazolo L5-a]pyrimidm-7-y )carbamate

[00413] Same procedure as [Example 97] . LCMS (M+l= 472)

Example 113. Synthesis of 3-(7-(cyclopropylamino)-3-formylpyrazolo[ 1 ,5-a]pyrimidin-5-yl)-N- methylbenzenesujfonamide

[00414] Same procedure as [Example 98] . LCMS (M+l= 372)

Example 114. Synthesis of 3 -(7-(cyclopropylammo)-3 -((2, 5 -dioxoimidazolidm-4- yiidcnc)nu:ih l )pyni/oio[ i .5 -a J p H n ¾ i 1 i -3 - \ s )-X~incU?yibcn/i:m:sairon nndc

00415] Same procedure as [Example 99] . LCMS (M+l= 454) Example 115. Synthesis of tert-buty 1 c clopropy 1(3 - forrny 1 -5 - (3 -

¾ methyisulfonamido)phenyl)pyrazolo [ 1 ,5 -a]pyrimidin-7-yl)carbamate

[00416] Same procedure as [Example 97] . LCMS (M+l= 472)

Example 116. Synthesis of N-(3 -(7-(cy cloprop lamino)-3 -form lpyrazolo [ 1 ,5 -a]pyrimidin-5 - yl)phenyl)methanesulfonamide

[00417] Same procedure as [Example 98] . LCMS (M+l= 372)

Example 117. Synthesis of N-(3 -(7-(cy cloprop lamino)-3 -((2,5 -dioxoimidazolidin-4- ylidene methyl)pyrazolo[l -a]pyrimidin-5-yl)phenyl)methanesulfonamide

00418] Same procedure as [Example 99] . LCMS ( M · ! 454) Example 1 18.

fom ylpyrazolo[l ,5-a]pyrimidin-7-yl)carbaniate

[00419] Same procedure as [Example 97] . LCMS (M+l= 422)

Example 1 19. Synthesis of 7-(cyclopropy lamino)-5 -{3 -(dimethylamino )phenyl)pyrazolo [1 ,5- a] pyri m idine-3 - carbal dehy de

[00420] Same procedure as [Example 98] . LCMS (M+l= 322)

Example 120. Synthesis of 5 -((7-(cy clopropylamino)-5 -(3 -(diniethy lamino)phenyl)-

00421] Same procedure as [Example 99] . LCMS ( M · ! 404) Example 121.

vKeyclopropyQearbamate

[00422] Same procedure as [Example 97] . LCMS (M+l= 404)

Example 122. Synthesis of 3-{7-(cyclopropylammo)-3-fomiylpyrazolo[l ,5-a]pyrimidin- 5- yl)benzonitrile

[00423] Same procedure as [Example 98] . LCMS (M+l= 304)

Example 123. Synthesis of 3 -(7-(cyclopropylamino)-3 -((2, 5 -dioxoimidazolidin-4-

00424] Same procedure as [ Example 99] . LCMS (M+l Example 124. Synthesis of tert-buty l cyclopropyl(5 ~(3 -Pi uorophenyl)-3 -f rmy Ipyrazolo [ 1 ,5 - a]pyriniidin-7-yl)carbamate

[00425] Same procedure as [Example 99] . LCMS ( M · ! 397) Example 125. Synthesis of 7-(cyclopropylamjno)-5-(3-fluorophenyi)pyrazolo[ 1 ,5-a]pyrirmdine- 3-carbaldehvde

[00426] Same procedure as [Example 98] . LCMS (M+l= 297)

Example 126. Synthesis of 5-((7-(cyclopropylamino)-5-(3-fluorophenyl)pyrazolo[l ,5- aJp> ^¾"'^ΓΓ!"^!;^^η--^- '^ >n^~'¾i s k?^p _:c) r_!H^' ia/olliiir;c-2. -¾.iit¾iu:

00427] Same procedure as [Example 99] . LCMS (M+l= 379) Example 127. S nihesis tti^" eri-hitf y j c \ c 11 ¾ p i ¾ ^ p \ 1 { - { > r t r; \ I - - { ; ΐ \ i i 11 i t ¾ - 3 - s 1 sp ra/oio[ 1 .5- a pyrimidin-7-yl)carbamate

[00428 J Same procedure as [Example 97] . LCMS (M+l = 380) Example 128.

carbaldehydc

[00429] Same procedure as [Example 98] . LCMS (M+l= 280)

Example 129. Synthesis of 5 -((7-(cyclopropy lamino)-5 -(pyridin-3 -yl)pyrazolo [ 1 ,5 -a.]pyrimidin- 3-yl)methylene imjdazolidine-2,4-dione

00430] Same procedure as [Example 99] . LCMS ( M · 1 362) Example 130.

a]pyriniidin-7-yl)carbamate

00431] Same procedure as [Example 97] . LCMS (M+l= 380)

Example 131. Synthesis of 7-(cyclopropy lamino)-5 -(pyridm-4-yl)pyrazolo [ 1 , 5 -alpyrimidine-3 - carbaldehyde

32] Same procedure as [Example 98] . LCMS (M+l=

Example 132. Synthesis of 5-((7-( ^'cyclopropylamino)-5-(pyridin-4-yl)pyrazolo[ 1 ,5-a]pyrimidin- 3-yj metjwlene)imidazojidme-2,4-dione

Same procedure as [Example 99] . LCMS (M+l= 362) Example 133.

formylpyrazoloj^" 1 ,5-a]pyrimidin-7-yl)carbamate

[00434] Same procedure as [Example 97] . LCMS ⁽ hi^■ ! 398)

Example 134. Synthesis of 7-(cyciopropylaminoV5-(2-fluoropyridin-4-yl)pyrazoio[l ..5- a]pyrimidine-3-carbaldeh de

[00435] Same procedure as [Example 98] . LCMS (M+l= 298)

Example 135. Synthesis of 5-((7-(cyclopropylamino)-5-(2-fluoropyridiii-4-yl)pyrazolo Γ1 -.5- a sp nnsiiiin-3-y] jmeihs knc );mida/olidinc~2.4-d:onc

00436] Same procedure as [Example 99] . LCMS (M+l Examp

[00437] Same procedure as [Example 97] , LCMS ( M · ! 395)

Example 137, Synthesis of 7-(cycjopropylamjno)-5-{4-hydroxyphenyl)py

a]pyrimidine-3-carbaldehyde

[00438] Same procedure as [Example 98] . LCMS (M+ l= 295)

Example 138, Synthesis of 5-(( 7-f cyclopropylamino )-5-(4-hydroxypheny[)pyrazolo[ 1 ,5- a1pyrimidin-3-yl.)rn.elhylene)irnidazolidine-2.4-dione

00439] Same procedure as [Example 99] , LCMS (M+l Example 139. Synthesis of tert-buty 1 cy clopropy 1(3 - forrny 1 -5 - (3 -

(morpholinomethyl)phenyl)pyrazolo[l,5-alpyrimidin-7-yl carbamate

[00440] Same procedure as [Example 97] . LCMS (M+l Example 140. Synthesis of 7-(cyclopropylarriino)-5-(3-

(rn rphoiiniinieihs Hphcnyi ;pvra/.oioj j .5-a ipvnm;dine~3-au†>aidchydc

[00441] Same procedure as [Example 98] . LCMS (M+l= 378)

Example 141. Synthesis of 5-((7-(cyclopropylamino)-5-(3-

(morpholinom.ethyl)phenyl)pyrazolo[1 ,5-ajpyrHnidin-3-yj)m-ethylene)imidazojidine-2, dione

00442] Same procedure as [ Example 99] . LCMS (M+l 142. Synthesis of tert-butyi- cy

)methyl)phenyl)pyrazolo[^"L5-a]pyrimidm-7-y )carbamate

[00443] Same procedure as [Example 97] . LCMS ( M · ! 491 )

Example 143. Synthesis of 7-(cyclopropylamino)-5 -(3 -(¾ 4-methylpiperazin- 1 - yl)methyl)plienyl)pyrazolo[l,5-a]pyrimidine-3-carbaldehyde

[00444] Same procedure as [Example 98] . LCMS ( M^■ i 391 )

Example 144. Synthesis of 5-((7-(cyclopropylamino)-5-(3-{{4-methy

yl)methyl)phenyl)pyrazolori,5-a]pyrimidin-3-yl)methylene)imi

00445] Same procedure as [Example 99] . LCMS (M+ l= 473) Examp

[00446] Same procedure as [Example 97] . LCMS (M+l= 450)

Example 146. Synthesis of N-( 3 -(7-(cyclopropylamino)-3 -formylpyrazolo j^" 1 ,5 -a]pyrimidin-5 - vDbenzyDacetamide

[00447] Same procedure as [Example 98] . LCMS (M+l= 350)

Example 147. Synthesis of N-(3-(7-(cyclopropylamino)-3-((2,5-dioxoirnidazolidin-4- ylidene)meth l)pyrazolo[l ,5-a]pyrimidin-5-yl)benzyj)acetamide

00448] Same procedure as [Example 99] , LCMS ( M · ! 432) Example 148. Synthesis of 5-{3-{aminomethyl)phenyl)-7-{eyelopro

a]pyrimidine-3-carbaldehyde

[00449] To tert-butyl 5-(3-(acetamidomethyl)phenyl)-3-forniylpyrazolo[ 1 ,5-a]pyrimidin-7- yl(cyclopropyi)carbamate (50 mg, 0, 1 11 mmol) was added 1 mL of 4M HCl in 1,4-dioxane and 1 mL of FLO. The reaction mixture was stirred at 80° C for 16 hours then cooled to room temperature and diluted with H?0, To the reaction mixture, 5M NaOH was added to adjust pH to >10 then the mixture was extracted with CH₂CI₂. The organic layer was collected, dried over MgS0_4> filtered and evaporated to dryness to provide 24 mg of 5-(3-(arrrinomethyl)phenyl)-7- (cyclopropylamino)pyrazolo[l ,5-a]pyrimidine-3~carba!dehyde (70%). LCMS (M+l. ==308)

Example 149. Synthesis of 5-((3-(. aminomci yi ¾phcriyi -7-{cyclppnspyiarninp)pyra/plo! 1 .5- alpyrimidin-3-yl)methylene)imidazolid.ine-2,4-dione

[00450] Same procedure as [Example 99] . LCMS (M ! 390)

[00451] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 98 and Example 99. All compounds were

characterized by LCMS. Table 22B shows the biological activities of the compounds listed in Table 22A.

Table 22A.

ı76

Table 22B,

CK2: IC50 ΡΪΜ2: IC50 AB: MDAMB453 AB: BxPC3

Compound

(5 uM ATP) (μΜ)

R7 <0.01 > 2.5000 1.054 23.617

S7 <0.01 > 2.5000 1.174 11.78

T7 <0.01 > 2.5000 1.298 6.592

1, 7 <0.01 > 2.5000 1.153 1.191

V7 <0.01 > 2.5000 1.964 14.486

VV7 <0.1 > 2.5000 0.683 1 .898

X7 <0.01 0.867 4.746 > 30

Y7 <0.1 1 .3082 1.938 2.578

Z7 <0.01 1.4748 1.79 0.725

A8 <0.01 1.2497 > 30 14.347

B8 <0.01 > 2.5000 > 30 12.535

C8 <0.01 > 2.5000 17.123 1.,232

D8 <0.01 0.0754 5.276 0.549

E8 <0.01 > 2.5000 11 ,733 > 30

F8 <0.01 0.2562 1 ,068 0.745

G8 <0.01 0.0487 14,882 10.61

H8 <0.01 > 2.5000 20.012 4.608

18 <0.1 > 2.5000 1.706 2.744

J8 <0.01 > 2.5000 1.263 8.129

K8 <0.1 > 2.5000 12.417 > 30

L8 <0.01 2.084 12.278 > 30

M8 <0.01 1 777] > 30 > 30

N8 <0.01 > 2.5000 1.979 2.253

08 <0.01 > 2.5000 15,69 29.035

P8 <0.01 0.948 1.742

Q8 <0.01 > 2.5000 26.74 5.426 8 <L0 > 2.5000 Example 150. Synthesis of methyl 5-{7-(tert-butoxyc3rbony j {cyclopropyl)a.nimo)-3- forn ylpyrazolo[l,5-a]pyrimidin-5-yl)thiophene-2-carboxylate

[00452] Same procedure as [Example 97] . LCMS (M+l= 443)

Example 151. Synthesis of methyl 5-(7-(cyclopropylamino)-3-formylpyrazolo[ 1 ,5-a.]pyrimidin- 5 -y j)thiophene-2-carboxy 1 ate

[00453] Same procedure as [Example 98] . LCMS (M+l = 343)

Example 152. Synthesis of m ethyl -(7-(cy c I opropy lam in o)-3 -({2.5 -dioxoimi dazoiidi n-4- ylidene)methyl)pyrazolo[l,5-a]pyrimidin-5-yl)thiophene-2-carboxylate

00454] Same procedure as [Example 99] . LCMS (M+l= 425) Example 153.

a]pyrimidin-7-yl(cyclopropyl)carbamate

[00455] Same procedure as [Example 97] . LCMS (M+l= 410)

Example 154. Synthesis of 5-(7-( cyclopropylammo)-3 -formylpyrazolo [ 1 ,5-a]pyrimidin-5- yl)thiophene-2-carbonitrile

[00456] Same procedure as [Example 98] . LCMS ( IV! I 310)

Example 155. Synthesis of 5 -(7-f cy cl opropylarmnoV 3 -((2 , 5 -dioxoimidazolidin-4- ylidene)methyl)pyrazolo[l ,5-a]pyrirrndin-5-yl)thiophene-2-carbon^

00457] Same procedure as [Example 99] . LCMS ( M · ! 392) Example 156. Synthesis of 5-{7-{tert-butcxycarbori3i.{cyck^propyj)amino)-3- formylpyrazolo[l,5-aJpyrimidin-5-yl)thiophene-2-carboxylic acid

00458J To tert-butyl 5-chloro-3~forniylpyrazolo[l ,5-a]pyrimidin-7-yl

(cyclopropyl)carbamate (1 g, 2.97 mmol) in 30 mL of a 2: 1 mixture of 1,2-dimethoxyethane/ EtOH was added 2-carboxythiophene-5- boronic acid (766 mg, 4.45 mmol),

tetrakis(triphenylphosphiiie)palladmm(0) (171 mg, 0.148 mmol), and 2M aqueous solution of Na₂C0₃ (4.45 mL, 8.91 mmol). The mixture was stirred at 95°C for 3 hours then cooled to room temperature and partitioned between 2N NaOH and ethyl acetate. The layers were separated and the aqueous layer was acidified to pH<3 with cone. HC1. The aqueous layer was extracted (3x) with methylene chloride. The combined organic layers was washed with brine, dried oyer MgS0₄, filtered, and evaporated to dryness to provide 450 mg of 5-(7-(tert- butoxycarbonyl(cyclopropy[)amino)-3-fom y[pyrazolo[l,5-a]pyrimidin-5-yl)thiophene-2- carboxylic acid. Some additional material which was in the first ethyl acetate layer was purified by silica gel chromatography (0%-2()% MeOH/CH₂Cl₂) to provide another 550 mg of 5-(7-(tert- biitoxycarbonyl(cyclopropyl)amino)-3-formylpyrazolo[l,5-a]pyrimidin-5-yl)thiop

carboxylic acid (79%). I . CMS (M · 1 429)

Example 157. Synthesis of 5-(7-(cyclopropylamino)-3-forrnylpyrazolo[ 1 ,5-a]pyrimidin~5~

yl)thiophene-2-carboxylic acid

[00459J To 5-(7~(tert-butoxycarbonyl(cyclopropyl)am

5-yi)thiophene-2-carboxylic acid (1 g, 2.33 mmol) was added 8 mL of 4M HQ in dioxane and another 5 mL of dioxane, The reaction mixture was stirred at 80° C for 2 hours, cooled to room temperature and partitioned between CLLCL and ¾0. The emulsion that formed between the layers was filtered off^" and rinsed with ¾0. The recovered solid was dried under vacuum to provide 627 mg of 5-(7-(cyclopropylamino)-3-fonnylpyrazolo[l ,5-a]pyrinTidm-5-y[)thiophene- 2-carboxylic acid as a red solid (82%). LCMS (M+ 1=329)

Example 158.

(3-methox ropyl)thiophene-2-carboxamide

[00460] To 5-(7-(cyciopropylarnino)-3-fomiylpyrazolo[l ,5-a]p3_'Timidin-5~y

carboxylic acid (30 mg, 0,091 mmol), EDCI (19 mg, 0.10 mmol), Et₃N (14 μΕ, 0, 10 mmol), and HOBt (14 mg, 0.10 mmol) in 2 mL of DMF pre-stirred for 5 minutes was added 3- methoxypropylamine (10 μΕ, 0.10 mmol). The reaction mixture was stirred at room

temperature for 1 hour. The reaction was diluted with ethyl acetate, washed with H₂0, brine, dried over MgS0₄, filtered, and evaporated to dryness to provide 30 mg of 5-(7- (cyclopropylamino)-3 -formylpyrazolo [ 1 ,5 -ajpyrimi din-5-yl)-N-(3-methoxypropyl)thiophene-2- carboxamide (83%). LCMS (M+l=400)

Example 159. Synthesis of 5-(7-(cyclopropyiammo)-3-((2,5-dioxoimidazolidin-4- yiidene niethyl)pyrazolo[L5-a]py^{^}

carboxamide

[00461] To 5-(7-(cyclopropylamino)-3-formylpyrazo!o[ 1 ,5-a]pyrimidin-5-yl)-N-(3- methoxypropyi)thiophene-2-carboxamide (30 mg, 0.075 mmol) in EtOH (1 mL) was added piperdine (20 μΕ, 0.150 mmol), and hydantoin (10 mg, 0.075 mmol). The reaction mixture was stirred at 85°C for 3 hours. The solid formed was isolated by filtration to provide 5-(7- (cyclopropylamino)~3-((2,5-dioxoiim

yl)-N-(3-methoxypropyl)thiophene-2-carboxamide. LCMS (M+l=482)

Example 160. Synthesis of 5-(7-¾ eyclopropylammo)-3-formylpyrazolo[ 1 ,5-a]pyrimidin-5- yl)thiophene-2-carboxamide

[00462] To 5-(7-(cyclopropylamino)-3-formylpyrazolo[ 1 ,5-a]pyrimidin-5-yl)thiophene-2- carboxylic acid (40 mg, 0.122 mmol), HATU (70 mg, 0.183 mmol), HOBt (4 mg, 0,024 mmol) and DIEA (85 ₍uL, 0.488 mmol) in 2 mL of DMF was added ammonium chloride (20 mg, 0.366 mmol). The reaction mixture was stirred at room temperature for 1 hour. The reaction rmxtue was diluted with ethyl acetate washed with saturated NaHC0₃ solution, brine, dried over MgS04, filtered, and evaporated to dryness to provide 42 mg of 5-(7-(cyclopropylamino)-3- formylpyrazolo [ 1 ,5-a]pyrimidin- 5 -yl)tbiophene-2-carboxamide ( 100%) . LCMS (M+ 1 =328) .

Example 161. Synthesis of 5 -(7-(cycl opropylaminoV 3 -((2 , 5 -dioxoimidazolidin-4- yiidene)methyl)pyrazolo[l ,5-a]pyrimidin-5-yl)thiophene-2-carboxamide

[00463] Same procedure as [Example 159] . LCMS (M+l=410)

[00464] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 158 and Example 159. Ail compounds were characterized by LCMS, Table 23B shows the biological activities of the compounds listed in

Table 23 A.

ı94

Table 23Λ.

Table 23B,

CK2: IC50 ΡΪΜ2: IC50 AB: MDAMB453 AB: BxPC3

Compound

(5 uM ATP) (μ )

D9 <0.01 > 2.5000 0.869 1.983

E9 <0.01 > 2.5000 0.397 0.496

F9 <0.01 > 2.5000 0.312 0.643

G9 <0.01 > 2.5000 0.31 0.657

H9 <0.01 > 2.5000 0.251 10.512

19 <0.01 0.7137 > 30 > 30

J9 <0.01 > 2.5000 0.795 1.736 9 <0,01 > 2.5000 9.378 11.666

L9 <0.01 > 2.5000 2.066 3.829

M9 <0.01 > 2.5000 1.266 1.469 9 <0.01 1.134 5.413

09 <0.01 0.621 12.558

P9 <0.01 0.596 0.5

Q9 <0.01 1.044 2.134

R9 <0.01 1.554 1.555

S9 <0.01 5.882 5.532

T9 <0.01 0.444 0.956

U9 <0.01 1.479 4.863

V9 <0.01 1.567 2.905

W9 <0.01 1.145 0.885

X9 <0.01 > 2.5000 1.391 > 30

Y9 <0,01 > 2.5000 0.389 0.438

Z9 <0.01 > 2.5000 0.762 1.337

AlO <0.01 > 2.5000 0.408 2.115

BIO <0.01 > 2.5000 0.895 1.167

CIO <0.01 0.7939 0.66 2.399

D10 <0.01 > 2.5000 1.529 6.508

E10 <0.01 > 2.5000 0.557 0.624

FI O <0.01 > 2.5000 0.251 0.323

GI O <0.01 > 2.5000 1.038 0.995

H10 <0.01 > 2.5000 0.294 2.968 CK2: IC50 ΡΪΜ2: IC50 AB: MDAMB453 AB: BxPC3

Compound

(5 uM ATP) (μ )

110 <0.01 > 2.5000 0.813 1.386

J10 <0.01 > 2.5000 0.613 0.324 10 <0.01 > 2.5000 0.579 0.451

L10 <0.01 > 2.5000 2.275 0.792

M10 <0.01 1.7758 5.94 0.677

N10 <0.01 > 2.5000 0.958 0.455

010 <0.01 1.8944 0.537 0.297

P10 <0.01 > 2.5000 0.394 0.451

QIO <0.01 > 2.5000 1.782 16.637

RIO <0.01 > 2.5000 1.641 5.729

S 10 <0.01 > 2.5000 10.053 18.645

T10 <0.1 > 2.5000 > 30 >30

U10 <0.01 > 2.5000 13.297 21.203

VI 0 <0.01 2.1321 > 30 >30

W10 <0,01 1 .3653 0.236 0.63

X10 <0.01 > 2.5000 0.937 0.917

YIO <0.01 > 2.5000 0.79 > 30

Z10 <0.01 > 2.5000 2.336 22.798

Al l <0.01 > 2.5000 0.458 0.724

Bl l <0.01 > 2.5000 >30 1.262

Cl l <0.01 > 2.5000 27.783 3.302

Dl l <0,01 > 2.5000 1.445 2.265

El l <0.01 > 2.5000 1.298 2.948

Fl l <0.01 > 2.5000 0.567 0.903

Gi l <0.01 2.0441 0.231 0.494

Hl l <0.01 > 2.5000 1.11 2.705

111 <0.01 > 2.5000 1.232 0.591

Jl l <0.01 > 2.5000 0.833 1.234

Kl 1 <0,01 > 2.5000 0.546 1 ?5?

Ll l <0.01 > 2.5000 1.004 0.816

Mi l <0.01 > 2.5000 1.016 0.745 CK2: IC50 ΡΪΜ2: IC50 AB: MDAMB453 AB: BxPC3

Compound

(5 uM ATP) (μ )

Ni l <0.01 > 2.5000 1.266 2.261

Oi l <0.01 > 2.5000 0.887 4.986

Pl l <0.01 > 2.5000 0.487 0.517

Ql l <0.01 > 2.5000 0.621 0.564

R] 1 <0.01 > 2.5000 0.845 2.309

SI 1 <0.01 > 2.5000 1.935 >30

Ti l <0.01 > 2.5000 0.193 >30

Ul 1 <0,01 > 2.5000 0.618 5.349

VI 1 <0.01 > 2.5000 0.892 1.6

Wl l <0.01 > 2.5000 0.156 3.435

Xl l <0.01 1 .7245 3.806 0.225

Yl l <0.01 > 2.5000 1.402 0.352

Zl l <0.01 1 .2434 2.251 0.355

A12 <0.01 1.4396 1.151 0.445

B12 <0,01 > 2.5000 0.399 2.764

C12 <0.01 > 2.5000 >30 >30

D12 <0.01 > 2.5000 0.683 0.854

E12 <0.01 > 2.5000 29.518 2.348

F12 <0.01 > 2.5000 >30 > 30

G12 <0.01 0.8106 0.658 0.352

H12 <0.01 > 2.5000 0.449 0.418

112 <0.01 > 2.5000 1.282 1 .516

J12 <0.01 > 2.5000 0.52 0.94

K12 <0.01 1.872 1.338 0.379

L12 <0.01 > 2.5000 0.498 > 30

M12 <0.01 1.2604 7.403 8.736

N12 <0.01 3.52 > 30

012 <0.01 > 2.5000 1.077 2.509

P12 <0.01 > 2.5000 1.014 3.421

Q12 <0.01 > 2.5000 0.942 7.084

R12 <0.01 > 2.5000 0.846 14.096 CK2: IC50 ΡΪΜ2: IC50 AB: MDAMB453 AB: BxPC3

Compound

(5 uM ATP) (μ )

S 12 <0.01 > 2.5000 1.034 4.897

T12 <0.01 > 2.5000 0.767 2.662

U12 <0.01 > 2.5000 0.525 > 30

V12 <0.01 1.759 > 30

W12 <0.01 1 .041 1.184

X12 <0.01 7.54 > 30

Y12 <0.01 0.692 1.706

Z12 <0,01 2.17 9.892

A13 <0.01 > 2.5000 0.534 0.996

B13 <0.01 > 2.5000 0.388 2.584

Example 162. Synthesis of 4-f7-(tgrt-b^tox■Vcarbonyl■(^'cvclo ro yl amino -3- fon^·n lpyrazolo[l _<5-g]pyrimidin-5-yl)thiophene-2-carboxylic acid

[00465] Tert-b yl 5 -chloro-3 -form lpyrazolo [ 1 ,5 -a]pyrimidine-7-yl)(cyclopropyl)carbamate (0.5 g, 1 .48 mmol) and commercially available (Combi-Blocks) 2-carboxythiophene-4-boronic acid pinacol ester (754 mg, 2.97 mmol) were dissolved in acetonitrile. 2M Na.₂C0₃ (1 mL) was added and the solution was degassed with a stream of N₂ for 10 min. PdCl₂dppf-CH₂Cl₂ (60 mg, 0.07 mmol) was added and the reaction was heated to 100 °C for 1.5 h. The solution was diluted with 1 ,5N NaOH (80 mL) and filtered over celite. The pH of the filtrate was adjusted to pH^:::3 by the addition of 6M HCl. The resulting precipitate was filtered and dried in vacuo to afford 4- (7-(fer/-butoxycarbonyi(cycloprop

2-carboxylic acid (473 mg, 74%) as a tan solid. LCMS (ES): >90% pure, mJz 429 [M+l]⁺. Example 163. S niiscsis t ij^" -1-{ 7-(cyck>prop i jiins no $-3-f rsin ips razt iju 1.5-</j j s riii¾uiivi-3- yl^")thiophene-2-earboxylic acid

[00466] 4-(7-( rert-butoxycarbonyl(cyclopropyl)amino)-3 -formylpyrazolo [1 ,5 - ]pyrimidin-5 - yl)thiophene-2-carboxylic acid (473 mg, 1.10 mmol) was dissolved in dichloromethane (5 niL) and trifluoroacetic acid (3 mL). After 1 h, the dark red solution was concentrated under a stream of air. The red oil was triturated with I¾0 (5 mL) and the precipitate was filtered to provide 4- (7-(cyclopropyl)amino)-3 -formylpyrazolo [1,5 - ]pyrimidin-5 -yl)thiophene-2-carboxylic acid (321 mg, 88%). LCMS (ES): >95% pure, m/z 329 [M+l] ^÷. Example 164. Synthesis of (2 -4-(7-(cyclopropyl)am o)-^^

ylidene)methyl)pyrazolo[l,5-a]pyrimidin-5-yl)thiophene-2-carboxy^ acid

[00467] Hydantoin (292 mg, 2.92 mmol) and piperidine (285 xL, 2.89 mmol) were added to 4-(7-(cyclopropy3)ammo)-3-formy3pyrazolo[l acid (315 mg, 0.96 mmol) dissolved in ethanol (5 mL). The reaction was heated at 80 °C. After 15 h, the reaction was cooled to r.t., then diluted with water (10 mL). The pH was adjusted to pH=3 by addition of IN HQ. The yellow precipitate was collected and washed with 1 : 1 ethanohwater (10 mL) and then ethanol (10 mL). The solid was dried in vacuo to give (Z)-4-(7- (cyclopropyl)amino)-3-((2,5-dioxoimidazoiidm

yl)thiophene-2-carboxylic acid (362 mg, 92%). LCMS (ES): >95% pure, m/z 411 [M+l]⁺. Example 165. Synthesis of (Z)-5-((7-(cy

carbonyl)thiopheii-3-yl)pyrazol^^

dione

[00468] (Z)-4-(7-(Cyclopropyi)arnino)-3-((2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazo[o[l,5- ]pyriniidm-5-y[)thiophene-2-carboxy[ic acid (1.0 eq, 34 mg, 0.0828 mmol) was mixed in a vial with HOBt.H₂0 (2.0 eq, 22 mg, 0.163 mmol), 2,6

dimethylmorpholine (isomer mixture, 4.0 eq, 41 ul, 0.333 mmol), DIEA (2.0 eq, 29 ul, 0.166 mmol) in NMP (0.5 ml). EDCI (2,0 eq, 32 mg, 0.166 mmol) was added and the mixture was stirred at 70°C for 1 hour. Water was added and the resulting precipitate was filtered and dried. The material was triturated in a mixture of ethyl acetate and hexanes. filtered and dried in vacuo to give (Z)-5-((7-(cyclopropylarnino)-5-(5-(2₅6-dimethylmoTpholine-4-carbonyl)thiophen-3- yl)pyrazolo[l,5-a]pyrimidm-3-yl)methylene as a yellow solid (26 mg,

62% yield). i .C MS (ES): >95% pure, m/z 508 [M+lf.

[004691 The following compounds were prepared using conditions similar to the chemistries described in Example 165. All compounds were characterized by LC S. Table 24B shows the biological activities of the compounds listed in Table 24 A.

Table 24 A.

Table 24B.

Example 166. Synthesis of 5 -i ydroxynicihyi )ihiophcn-2-boroi:j e acid

[00470 J 5-(Hydroxymethyl)thiophen-2-boronic acid was prepared from the commercially available 5-formylthiophen-2-boromc acid (Combi-Blocks) according to the procedure described in patent application WO2007/1 1 8137.

Example 167. Synthesis of tert-butyl cyclopropyl(3-forniyl-5-(5-(hydroxymethyl)thiophen-2- yl) yrazolo[ 1 ,5- ]pyrimidine-7' ^■yl)carbamate

[00471 J Note: DME and 2M Na₂C0₃ were degassed with a stream of N₂ in separate flasks prior to addition. Tert-butyl 5-chIoro-3-formylp>Tazolo[l,5- ]pyrimidine-7- yl)(cyclopropyl)carbamate ( 1.5 g, 4.45 mmol) was dissolved in DME: (40 mL). Crude 5- (iiydroxymethyl)tiiiopiien-3-boronic acid (1.4 g, 8.9 mmol) was added, followed by Pd(PPh₃)₄ (510 rng, 0.45 mmol) and finally 2M Na₂C0₃ (6.7 mL, 1 3.3 mmol). The reaction was heated to 90 °C for 2 h. The solution was partitioned between EtOAc (100 mL) and 0.5N HCI ( 100 mL). The aqueous layer was extracted with EtOAc (2 x 75 mL). The organics were washed with brine (250 mL), dried over MgS0 , filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30-45% EtOAc/hexanes) and then triturated with hexanes (3 x 10 mL) to yield tert-butyl cyclopropyl(3-formy[-5-(5-(hydroxymethyl)thiophen-2- yl)pyrazolo[l ,5- ]pyrimidine-7-yl)carbamate (984 mg, 53%) as an off white solid. LCMS (ES): >95% pure, m/z 415 [M+l ]⁺. Example 168. S nihcsis of 5-f 3-(bro-no: ncjh j )l ioplu:ii-2-yi )-7-

(cyclopropylamino)pyrazolo l,5- ]pyrimidine-3-carbaldehyde

[00472] Hydrogen bromide (48% in water, 5 mL) was added dropwise to fert-butyl cyekypropy](3-fomiyl-5~(5-(hydroxyniethyl)thiophen-2-y

yl)carbamate (980 mg, 2.4 mmol) suspended in dichloromethane (5 mL). The solution immediately became dark brown and homogeneous upon addition. The reaction was heated to 40°C for 4 hours, then diluted with dichloromethane (10 mL). The liquid was decanted and the gummy residue was washed with dichloromethane (3 x 10 mL). The combined liquids were washed successively with sat. NaHC0₃ (20 mL) and brine (20 mL), and then dried over gS0 , filtered and concentrated in vacuo. The residue was triturated with hexanes and then purified via flash column chromatography (10-20% EtOAc/hexanes) to provide 5~(5~

(bromomethyl)thiophen-2-yl)-7-(cyclopropy[amino)pyrazolo[l,5-a]pyrimidine-3-carbaldehyde (300 mg, 34%) as a yellow solid. LCMS (ES): >95% pure, m/z 378 [M+lf . Example 169. Synthesis of (¾-5-((7~(cycloprop

2-yl)pyrazoio l,5-alpyrimidin-3-yl)methylene iniidazolidme-2,4-dione

[00473] Potassium carbonate (30 mg, 0.20 mmol) was added to 5-(5-(bromomethyl)thiophen- 2-yi)-7-(cyclopropylamino)pyrazolo[l,5- ]pyrimidine-3-carbaldehyde (25 mg, 0.07 mmol) dissolved in DMF (0.7 mL). Pyrrolidine (6 μΤ, 0,07 mmol) was added and the reaction was heated to 60 °C for 4 h. Water (3 mL) was added and the orange precipitate was filtered and dried in vacuo to give 7-(cyclopropylammo)-5-(5-(pyrrolidin-l -ylmethyl)thiophen-2- y])pyrazolo[l,5- ]pyrirnidine~3~carba]dehyde (13 mg, 54%) which was used without further purification, LCMS (ES): >85% pure, m/z 368 [M+l]^"\

[00474] Hydantoin (3 mg, 0,03 mmol) and piperidine (3 μ,Ε, 0.03 mmol) were added to 7~

(cy clopropyS ami no )-5 -(5 -( pyrroli di n- 1 -y 1 methy l)t hiophen-2-yl)pyrazolo[l,5-a]pyrirnidine-3- carbaldehyde (12 mg, 0.03 mmol) dissolved in ethanol (0.5 mL). The reaction was heated at 80

°C. After 15 h, the reaction was cooled to room temperature then diluted with water (3 mL).

The precipitate was collected and washed with 1 : 1 ethanohwater (3 mL) and dried in vacuo to furnish (Z)-5-((7-(cyclopropylammo)-5-(5-pyrrolidin-l-ylmethyl)thiophen-2-yl)pyrazoio[l,5- a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (2,8 mg, 9% over two steps). LCMS (ES): >95% pure, m/z 450 [M+lf .

[00475] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 168 and Example 169, All compounds were characterized by LCMS, Table 25 B shows the biological activities of the compounds listed in

Table 25A.

Table 25 A.

Table 25 B.

Example 170. Synthesis of 5-(hydrox metfayl)thiophcn-3-boronic acid

[00476] 5-(Hydroxymethyl)thiophen-3-boronic acid was prepared from the commercially available 5-formylthiophen-3-boronic acid (Combi-Blocks) according to the procedure described in patent application WO2007/118137.

Example 171. Synthesis of tert-butyl cyclopropyl(3-fom yl-5-(5-(hydbroxymethyl)thiophen-3-

Jote: DME and 2M Na₂C0₃ were degassed with a stream of 2 in separate flasks prior to addition. en-butyl 5-chloro-3-formyipyrazolo[l,5-a]pyrimidme-7- yl)(cyclopropyl)carbamate (750 mg, 2.22 mmol) was dissolved in DME (22 niL). Crude 5- (hydroxymethy!)thiophen-3-boronic acid (880 mg, 5.57 mmol) was added, followed by

Pd(PPh₃)₄ (256 mg, 0.22 mmol) and finally 2M Na₂C0₃ (3.3 mL, 6.60 mmol). The reaction was heated to 90 °C for 2 h. The solution was partitioned between EtOAc (100 mL) and 0.5N HC1 ( 100 mL). The aqueous layer was extracted with EtOAc (2 x 75 mL). The organics were washed with brine (250 mL), dried over MgS0₄, filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30-45% EtOAc hexanes) and then triturated with hexanes (3 x 10 mL) to yield te/ -butyl cyclopropyl(3-formyl-5-(5- (hydroxymethyl)tMophen-3-y^ (638 mg, 69%) as an off white solid. ^lH NMR (CDCl₃, 400 MHz) δ: 10.34 (s, IH), 8.55 (s, I H), 8.11 (d, H i, ,/ 1 .6 Hz), 7.76 (d, IH, J= 1.6 Hz), 7.18 (s, 1H), 4.93 (bs, 2H), 3.30 (dddd, 1H, J= 6.8, 6.8, 3.6, 3.6 Hz), 2.1 (bs, I H), 1.42 (s, 9H), 0.85-0.92 (m, 21 1 h 0.63-0.70 (m, 2H). LCMS (ES): >95% pure,

Example 172. Synthesis of 7-(cycloprop lamino)-5 -(5 -(hydroxymethyl)thiophen-3 - yl)pyrazolo[L5-a]pyrimidine-3-carbaldehyde 2,2,2,-trifluoroacetate

[00478] Γβ/t-butyl cyclopropy 1(3 -formyl-5 -(5 -(hy droxymethy l)thiophen-3 -yl)pyrazolo [1,5- ]pyrimidine-7-yl)carbamate (20 mg, 0.05 mmol) was dissolved in dichioromethane (0.5 mL) and trifluoroacetic acid (0.5 mL). After 1 h, the solution was concentrated under a stream of air. The residue was purified via preparative HPLC to furnish 7-(cyclopropylamino)-5-(5- (hydroxymethyl }thiophen-3-yl)pyrazoio[ 1 ,5- ]pyrimidine-3-carbaldehyde 2,2,2-trifluoroacetate (4.8 mg, 23%).

Example 173. Synthesis of 5-(5-(bromome thyl)thiophen-3-yl)-7-

(cycl opropylamino ipyrazo lo [ 1 , 5 -q]pyrimidme-3 -carbaldehyde

[00479] Hydrogen bromide (48% in water, 2.5 mL) was added dropwise to tert-butyl cyclopropyl(3 -formy [-5 -(5 -(hydroxymethyl)thiophen-3 -y i)pyrazolo [ 1 ,5 - ]pyrimidine-7- y])carbamate (561 mg, 1 ,35 mmol) suspended in dichioromethane (3,5 mL), The solution immediately became dark brown and homogeneous upon addition. The reaction was heated to 40 °C for 3 h, then diluted with dichloromethane (10 mL). The liquid was decanted and the gummy residue was washed with dichloromethane (3 x 10 mL). The combined liquids were washed successively with sat. aHC0₃ (20 mL) and brine (20 mL), and then dried over MgS0₄, filtered and concentrated in vacuo. The residue was triturated with hexanes and then purified via flash column chromatography (15-40% EtOAc/hexanes) to provide

5 -(5 -(bromomethyl)thiophen-3 -y l)-7-(cy clopropylamino)pyrazolo [ 1 ,5 - ]pyrimidine-3 - carbaldehyde (105 mg, 20%) as a yellow solid. ^f H NMR (CDC1₃, 400 MHz) 5: 10.26 (s, 1 H), 8.44 (s, 1H), 8.12 (d, i l l. ./ 1.6 Hz), 7.80 (s, 1H), 6.73 (s, 1H), 6.65 fbs, 1H), 4.80 (s, 2H), 2.81 (m, ! i ! ). 1.03-1.09 (m, 211 ). 0.84-0.89 (m, 11 1 ). LCMS (ES): >95% pure, m/z 378 [M+l .

Example 174. Synthesis of (Z)-5-((7-(cyclopropylamm

3-yl)pyrazolo l,5-alpyrimidin-3-yl)methylene imidazolidine

[00480] Potassium carbonate (30 mg, 0.20 mmol) was added to 5-(5-(bromomethyl)thiophen~ 3-yl)-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (25 mg, 0.07 mmol) dissolved in DMF (0.7 mL), Pyrrolidine (6 ,u.L, 0.07 mmol) was added and the reaction was heated to 50 °C for 1.25 h. Water (3 mL) was added and the orange precipitate was filtered and dried in vacuo to give 7-(i^clopropylamino)-5-(5-(pyrrolidiii-l-ylmethyl)thiophen-'3- yl)pyrazolo[l ,5- ]pyrimidine-3-carbaldehyde (13 mg, 54%) which was used without further purification. LCMS (ES): >85% pure, m/z 368 [M+lf.

[00481] Hydantoin (3 mg, 0.03 mmol) and piperidine (3 μ,Ε, 0.03 mmol) were added to 7- (cyclopropylamino)-5-(5-(pyrrolidin- 1 -ylmethyl)thiophen-3-yl)pyrazolo[ 1 ,5- ]pyrimidme-3- carbaldehyde (12 mg, 0.03 mmol) dissolved in ethanol (0.5 mL). The reaction was heated at 80 °C. After 15 h, the reaction was cooled to r.i, then diluted with water (3 mL). The precipitate was collected and washed with 1 : 1 ethanohwater (3 mL) and dried in vacuo to furnish (Z)-5-((7~ (cyclopropylamino)-5-(5-pyrrolidin- 1 -ylmethyl)thiophen-3-yl)pyrazolo[l,5-a]pyrimidin-3- yl)rneihylerie)imidazolidine~2,4-dione (2.8 mg, 9% over two steps). LC S (ES): >95% pure, m 450 [M+lf.

[00482] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 174. All compounds were characterized by LCMS. Table 26B shows the biological activities of the compounds listed in Table 26A.

Table 26 A.

Table 26B.

183] The chemistry depicted in Figure 3 can be used to prepare analogs 7 substituted by a methyl group. Commercially available boronie acid 1 can be reacted with tert-butyl 5- chloropyrazolo[l,5-a]pyriniidin-7-yl(cyclopropyl)carbamate 2 under Suzuki reaction conditions to for methyl ketone 3. This compound 3 can be reacted with various substituted amines 4 under reductive amination conditions such as the conditions described in US2007/244094 or reaction conditions described in E uropean Journal of Medicinal Chemistry, vol 32, 1997, 143-150 to prepare compounds 5, Compound 5 can be converted to aldehyde 6 under Vilsmeier conditions, Compound 6 can be converted to compound 7 by reacting with hydantoin and piperidine in ethanol.

[00484J The molecules in the following Table 27 can be prepared using similar chemistries.

Table 27.

Example 175. Synthesis of 6-bromo~N~cyclopropyiimi dazo [ 1 ,2-q] yrazin-8-amine

[00485] Diisopropylethylamine (2.4 mL, 13.62 mmol) and cyclopropy famine (943 [iL, 13.62 mmol) were added to commercially available (Ark Pharm, Inc.) 6,8-dibromoimidazo[l,2~ ajpyrazine (2.51 g, 9,08 mmol) dissolved in 2-propanol (9 mL), The solution was placed in an 80 °C oil bath. After 4.5 h, the vol.atil.es were removed in vacuo. The brown residue was partitioned between dichloromethane (50 mL) and water (50 mL), The organic layer was washed further with water (50 mL) and then brine (50 mL). The organic layer was dried over MgS0₄, filtered and concentrated in vacuo. The residue was purified via a filtration over a short plug of silica gel (40% EtOAc/hexanes) and the filtrate was concentrated in vacuo to afford 6- bromo-N-cyclopropylimidazo[ 1 ,2- ]pyrazin-8-amine (2.19 g, 95%) as a light brown solid. ^lH NMR (CDCL, 400 MHz) δ: 7.61 (s, 1H), 7.46 (d, 1 H, ,/ 1 .2 Hz ), 7.44 (d, 1H, J = 1.2 Hz ), 6.26 (bs, I I I I 3.02 (dddd, I I I ./ 7,2, 7.2, 7.2, 3.6 Hz), 0.89-0.95 (m, 2H), 0.64-0.69 (ra, 2H). I .CMS C HS ): >95% pure, in / 254 [M+l f.

Example 176. Synthesis of te/ -butyl 6-bromoimidazo[l ,2-a]pyrazin-8-yl(cyclopiOpyl)

carbamate

[00486] 6-Bromo-N-cyclopropy3.imidazo[l ,2-a]pyrazin-8-arnine (0.5 g, 1 ,98 mmol) was dissolved in dichloromethane (8 mL). Di-tert-butyl dicarbonate (733 mg, 3.35 mmol), DMAP (5 mg, 0.02 mmol) and pyridine (0.4 mL) were added sequentially. After 12 h, the solution was diluted with EtOAc (50 mL) and then washed sequentially with IN HCl (50 mL), IN NaOH (50 mL), and brine (50 mL). The organic layer was dried over MgS0₄, filtered and concentrated in vacuo. The residue was triturated with hexanes (5 mL) to yield tert-bafyl 6- bromoimidazo[ 1 ,2-a]pyrazin-8-yl(cyclopropyl) carbamate (337 mg, 48%) as an off white solid.

Ή NM (CDCI₃, 400 MHz) 5: 8.18 (s, 1 H), 7.78 (d, 1 H, J = 0.8 Hz ), 7.67 (d, 1 H, J = 0.8 Hz ), 3.25 (dddd, 1H, J= 6.8, 6.8, 3.6, 3.6 Hz), 1.20 (s, 9H), 0.78-0.86 (m, 2H), 0.71-0.77 (m, 2H). LCMS (ES): >90% pure, m/z 354 [M+l ]⁺.

Example 177.

carbaldehyde

[00487 J Phosphorus! V) oxychloride (3.9 mL, 42.68 mmol) was added dropwise to anhydrous DMF (16 mL) at 0 ^C'C. 6-bromo-N-cyclopropylimidazo[ 1 ,2- ]pyrazin-8-amine (900 mg, 3.56 mmol) was dissolved in anhydrous DM F (24 mL) and added over two minutes. The solution was place in an 85 °C oil bath for 5 h. The solution was cooled to 0°C and cone. HCl (30 mL) was added. The mixture was basified to pH=10 wv 3N NaOH (-175 mL). The mixture was extracted with dichloromethane (3 x 250 mL), and the organics were washed with brine (500 mL), The organic layer was dried over MgS0 , filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30% EtOAc/hexanes) to furnish 6-bromo-8-(cyclopropylamino)imidazo[ 1 ,2- ]pyrazine-3-carbaldehyde (490 mg, 49%).

(ES): >95% pure, rn/z 282 [M+l f .

Example 178. Synihcsis or /i.77-buiyl 6-bi iio-3- l nriyii:iiiii;i/ )[ l .2-</jpyi'a/inc-K- vKcyclopropyQcarbamate

[00488] Di-tert-butyl dicarbonate (1.16 g, 5.30 mmol) and DMAP (21 rng, 0.18 mmol) were added to a solution of 6-bromo-8-(cyclopropylamino)imidazo[l ,2- ]pyrazine-3-carbaldehyde (994 mg, 3,50 mmol) in dichloromethane (1 5 raL). After 2.5 h, the solution was partitioned between EtOAc (100 niL) and water (100 mL). The aqueous layer was further extracted with EtOAc (2 X 75 mL), The organics were washed with brine (250 ml.), dried over MgS0₄, filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30% EtOAc/hexanes) to provide tert-butyl 6-bromo-3-formylimidazo[l ,2- ]pyrazine-8- yl(cyclopropyl)carbamate (1.17 g, 87%) as a brown foam. ^lH NM R (CDC1₃, 400 MHz) δ: 10.05 (s, 1H), 9.42 (s, 1H), 8.37 (s, IH), 3.25 (dddd, 1H, J = 6.8, 6.8, 4.0, 4.0 Hz), 1.22 (s, 9H), 0.85- 0.90 (m, 2H), 0.69-0.75 (m, 2H). LCMS (ES): >95% pure, m/z 382 | M · 1 j .

Example 179. Synthesis of tert-butyl cyclopropyl(3-formyl-6-(3-

1rifluoromethoxy) henyl)imidazof 1 ,2-q]pyrazin-8-yl)carbamate

[00489J Ί ert- butyl 6-bromo-3-formylimidazo[ 1 ,2- ]pyrazme-8-yl(cyclopropyl)carbamate (130 mg, 0.34 mmol), 3-(trifluoromethoxy)phenyl boronic acid (105 mg, 0.51 mmol), 3M NaaCOs ( 1.1 mL, 3.4 mmol) and DME (4.5 mL) were combined. The solution was degassed with a stream of N? for 10 min. Pd(PPb.3)₄ was added and the solution was retluxed for 2 h. The solution w^ras partitioned between dichloromethane (25 mL) and water (25 raL). The aqueous layer was further extracted with dichloromethane (2 x 25 mL). The organics were washed with brine (50 niL), dried over MgS0 , filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30-45% EtOAc/hexanes) to provide tert-bu y\ cyc!opropyl(3-

bright yellow solid. LCMS ( ES): >95% pure, m/z 463 | M · i ] . Example 180. Synthesis of fert-buty cyclopropyl(6-(3-trifluoromethoxy)phenyl)imidazo l ,2- q^" jp yr az in- 8 - yl)c arb ama te

)] 7¾rf-butyl cyclopropyl(6-(3-trifluoromethoxy)phenyl)irnidazo[l,2-i?]pyrazin-8- yl)carbamate (77%) was synthesized in a manner analogous to Example 179. LCMS (ES): >95% pure, m/z 435 | M · 1 j .

Example 181. Synthesis of fcrt-butyl cyclopropyl(6-(3-fliioroplienyr)-3-formylimidazo[L2- ]pyr3zin-8- i)carbarn3te

[00491] Jert-butyl cyclopropyl(6-(3-iluorophenyl)-3-formyliniidazo[ 1 ,2-a]pyrazm-8- yl)carbamate (28%) was synthesized in a manner analogous to Example 183. LCMS (ES): >95% pure, m/z 435 [M+l]⁺.

Example 182. Syncthesis of tert-butyi cyciopropyl(3-formyl-6-(3-

[00492] Tert-b tyl 6-bromo-3-formylimidazo[ 1 ,2-a]pyrazine-8-yl(cyclopropyl)carbamate (100 mg, 0.26 mmol), 4-[3-(4,4₅5,5-tetraniethyl-l ,3,2-dioxaborolan-2-yl)benzyl]morpholine (1 18 mg, 0.39 mmol), 3M Na₂C0₃ (1.3 mL, 2.60 mmol) and DME (3.5 mL) were combined. The solution was degassed with a stream of N? for 10 min. Pd(PPh₃)₄ was added and the solution was reiluxed for 2 h. The solution was partitioned between dichloromethane (25 mL) and water (25 mL). The aqueous layer was further extracted with dic loromethane (2 x 25 mL). The organics were washed with brine (50 mL), dried over MgS0₄, filtered and concentrated in vacuo. The residue was purified via preparative TLC (5% MeOH/dichloromethane) to afford tert-buiyl cyclopropyl(3-formyl-6-(3-(morpholinomethyl)phenyl)imidazo[l,2- ]pyra^ yl)carbamate (60 mg, 48%). LCMS (ES): >95% pure, m/z 478 | M 1 1 .

Example 183. Synthesis of ter /-butyl eyelopropy 1 ( 3 -formy 1-6- ((trimethylsilyl)ethyn^

[00493] Triethylamine (912 \iL, 6.56 mmol) was added to tert-butyl 6-bromo-3- formylimida.zo[ 1 ,2- ]pyrazine-8-yl(cyclopropyl)carbamate (250 mg, 0.66 mmol) dissolved in anhydrous DMF (2,2 mL) in a 15 mL pressure tube. The solution was degassed with a stream of N₂ for 10 min. Trimethylsilylacetylene (927 iiL, 6.56 mmol), Pd(PPh₃)₄ (76 mg, 0,07 mmol), and copper(I) iodide (25 mg, 0.13 mmoi) were added and the reaction was sealed and heated to 65 °C for 24 h. The reaction was diluted with EtOAc (50 mL) and then washed with 10% brine (4 X 50 mL) and brine (50 mL). The organics were dried over MgS0₄, filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30% EtOAc/hexanes) to give ferf-butyl cyclopropyl(3-formyl-6-((ftimethylsilyl)ethynyl)imidazo[l ,2- ]pyrazin-8- yl)carbamate (186 mg, 71 %) as a brown foamy solid. LCMS (ES): >95% pure, m/z 400

[M÷l]⁺. Example 184. S nihcsis tii^' i v -biii \ cloprop iC 3-Π Γη ιν nsscsix lcih n 1 >imicla <^[ i ,2- g]pyr az in- 8 -y 1 )c arb amate

Tert-hutyl cyclopropyl(3-formyl-6-((phenylet^^

yl )carbamate (64%) was synthesized in a manner analogous to Example 183. LCMS (ES): >95% pure, m/z 403 [M+l ,

Example 185. Synthesis of tert-butyl cyclopropyl(6-ethynyl-3-fom ylimidazor 1 ,2- lpyrazin-8- yl)carbamate

00495] Potassium carbonate (86 mg, 0.63 mmol) was added to tert-butyl cyclopropyl(3~ formyl-6-((trimethylsilyl)ethynyl)iniidazo[l ,2-a]pyrazin-8-yl)carbaniate (50 mg, 0.13 mmol) dissolved in methanol (2.5 mL). After 2 h, the volatiles were removed in vacuo. The residue was partitioned between dichloromethane (10 mL) and water (10 mL), The aqueous layer was further extracted with dichloromethane (2 x 10 mL). The organics were washed with brine (30 mL), dried over gS0₄, filtered and concentrated in vacuo. The residue was purified via flash column chromatography (30% EtOAc/hexanes) to provide tert-butyl cyclopropyl(6-ethynyl~3- formylimidazo[l ,2- ]pyrazin-8-yl)carbamate (20 mg, 50%) as a yellow foamy solid. LCMS (ES): >95% pure, m/z 327 [M+l . Example 186. Synthesis of fe^

6-{3-trifluoromethoxy)phenyl)iniidazo l ,2-g]pyrazm-8-yl)carbamate

[00496] Hydantoin (33 mg, 0,33 mmol) and piperidine (33 μΐ.,, 0.33 mmol) were added to tert-butyl cyclopropyl(3-formyl-6-(3-trifluoromethoxy)phenyl)imidazo[l,2-a]pyrazin-8- yl)carbamate (50 mg, 0.11 mmol) suspended in ethanol (0.5 mL). The reaction was sealed and irradiated in the microwave at 80 °C for 12 h. The precipitate was filtered off and washed with ethanol (3 mL) to give (Z)-tert-hutyl cyciopropyl(3-((2,5-dioxoimidazolidin-4~ylidene)methyl)- 6-(3-trifluoroniethoxy)pheny!)imidazo[l ,2-<2]pyrazin-8-yl)carbaniate (1 8 mg, 30%) as a bright yellow solid. LCMS (ES): >90% pure, m/z 545 [M+l]⁺.

Example 187. Synthesis of 5-((8-cyclopropylammo)-6-(3-(trifluoromethoxy)phenyl)

[00497] le/ -butyi cyclopropyl(3-((2,5-dioxoiniidazolidin-4-ylidene)methy[)-6-(3- triiluorom.ethoxy)phenyl)imidazo[l ,2-a]pyrazin-8-yl)carbamate (15 mg, 0.03 mmol) was dissolved in dichloromethane (0.5 mL) and trifluoroacetic acid (0.5 mL). After 1 h, the solution was concentrated under a stream of air. The residue was purified via preparative HPLC to furnish (Z)-5-((8-cyclopropylaniino)-6-(3-(trit1uoromethoxy)phenyl) imidazo[l ,2-a]pyrazin-3- yl)methylene)imidazolidine-2,4-dione (0.9 mg, 8%). Example 188.

6-(phenylethynyl)imidazo[l ,2-glpyrazin-8-yl)carbamate

[00498] Hydantoin (24 mg, 0,24 mmol) and piperidine (24 μΐ.,, 0.24 mmol) were added to tert-butyl cyclopropyl(3-formy!-6-((pheivylethynyl)imidazo[] ,2- ]pyrazm-8~yl)carbamate (24 mg, 0.06 mmol) dissolved in ethanol (1 mL). The reaction was heated at S0°C for 12 h, and then cooled to r.t. The precipitate was filtered off and washed with ethanol (3 mL) to give (Z)-tert-butyl cyclopropyl(3-((2,5-dioxoimidazolidin-4-ylidene)methyl)-6- (pheny iethynyl)imidazo[ 1 ,2- ]pyrazin-8-y l)carbamate ( 12 mg, 43%) as an orange/yellow solid. LCMS (ES): >90% pure, m/z 485 [M+l]⁺.

Example 189. Synthesis of (Z)-5-((8-(cyclopiOpylamino )-6-(phenylethynyl)imid.3zo[ 1 ,2-

[00499] (Z)-7ert-butyl cyc[opropyl(3-((2,5-dioxoimidazolidin-4-ylidene)methyl)-6- (phenylethynyl)imidazo[ 1 ,2- ]pyrazin-8-yl)carbamate (12 mg, 0.03 mmol) was dissolved in dichloromethane (0.3 mL) and trifluoroacetic acid (0.3 mL). After 1 h, the solution was concentrated under a stream of air. The residue was triturated with Et₂0 and filtered to yield (2)-5 ~((8-( cyclopropy 1 amino)-6-( heny 1 ethynyl)imi dazo [ 1 ,2-a]pyrazin-3 - yl)methylene)imidazolidine-2,4-dioiie (6 mg, 63%) as a bright yellow solid. Example 190.

nioipholinomethyl)phenyl)imidazo[l _^^^

dione 2.2.2 -trill uoroacetate

Hydantoin (152 mg, 1.50 mmol) and piperidme (150 μΤ, 1.50 mniol) were added to tert-butyl cyclopropyl(3-fom yl-6-(3-(mo^holinomem^

yljcarhamate (60 mg, 0.13 mmol) dissolved in ethanol (1 mL). The reaction was heated at 80°C for 4 d, and then diluted with water (10 mL). The supernatant was decanted and extracted with dichloromethane (2 15 mL). The organics were washed with brine (30 mL), dried over MgS0₄, filtered and concentrated in vacuo to a yellow solid. LCMS (ES): >95% pure, m/z 560 | M i | .

[00501] The crude solid was dissolved in dichloromethane (0.5 mL) and trifluoroacetic acid (0,5 mL). A fter 1 h, the solution was concentrated under a stream of air. The residue was purified via preparative HPLC to furnish (Z)-5-((8-(cyclopropylamino)-6-(3- mo^holmomethyl)phenyl)im^ 2,2,2 trifluoroacetate (5.5 mg, 8% over two steps).

Example 191. Synthesis of (Z)-5-((8-cyclopropylamino)-6-(3-fluorophenyl imidazo[l ,2- q]pyrazin-3- l)methylene)imidazolidine-2,4-dione

[00502] Hydantoin (1 8 mg, 0.17 mmol) and piperidine (17 _,uL, 0.17 mmol) were added to fert-butyl cyclopropyl(6-(3-fluorophenyl)-3-formylirnidazo[l ,2-a]pyrazin-8-yl)carbamate (23 mg, 0.06 mmol) dissolved in ethanol (0.3 mL). The reaction was heated at 80 °C for 18 h, and then concentrated in vacuo to a yellow solid. The crude solid was dissolved in dichloromethane (0,5 mL) and trifluoroacetic acid (1.5 niL). After 1 h, the solution was concentrated under a stream of air. The residue was triturated with ethanol and filtered to provide (Z}-5-((8~ cyclopropylamino)-6-(3-fluorophenyl)imidazo[l ,2- ]pyrazin-3-yl)methy

dione as an orange/yellow solid (2.4 nig, 10% over two steps).

Example 192. Synthesis of Related Compounds.

[00503] The compounds in the following table were prepared by the methods described above, by selecting appropriate starting materials as is apparent to the person of ordinary skill. Table 28B shows the biological activities of the compounds listed in Table 28A.

Table 28A.

Table 28B. CK2: IC50 ΡΪ 2: IC50 ΑΒ: MDAMB453 ΑΒ: BxPC3

(μ ) (5 μΜ ΑΤΡ) (μΜ) (μΜ)

C14 > 5.0000 > 2.5000

D14 <0.1 > 2.5000 4.531 3.69

E14 > 5.0000 > 2.5000

F14 > 5.0000 > 2.5000

G14 > 5.0000 > 2.5000

[00504] The chemistries described on Figure 7 can be used to prepare analogs substituted by a trifluoromethyl group. Commercially available 2~ammo-3,5-dibromopyrazme and commercially available 3-bromo-l,l,l-trifluoroacetone can be reacted together at 50°C in a solvent such as dioxin ( conditions previously described in WO2003/82817), to prepare compound 3 Compound 3 can be reacted with amine RjNH? to obtain 4. This material can be protected by a boc group by reacting 4 with a reagent like Boc₂0 to obtain 5. This material can be further transformed into 6 under viismeir conditions in the presence of POCI₃. Compound 6 can be reacted with various reagents such as boronic acids or esters W-B(OR₃)₂ under Suzuki conditions to form molecule 7.

[00505] Other analogs of 7 can be prepared by heating 6 with amines or anilines _{5 6} H, alcohols or phenols R₅OH, thiols or thiophenols R₅SH, in the presence of a base or an acid. Compound 8 can be prepared by hea ting 7 with hydaiitoin in ethanoi in the presence of a base such as piperidine. GENERAL METHODS

[00506] Unless otherwise specified, the various substituents of the compounds are defined in the same manner as the formula II / IF compound of the invention.

[00507] The chemistry described in Figure 4 and Figure 5 can be used to prepare various substituted compounds of formula II.

[00508] Substituted aminopyrazole 1 can react with isothiocyanate 2 to form intermediate 3. Compound 3 can be cyclized to 4 in the presence of a base such as sodium hydroxide.

Compound 4 can be alkylated by with RTFl alo in the presence of a base. Compound 5 can be converted to compound 6 using phosphorus oxychloride. Molecule 7 can be prepared by addition of amine R₇RgNH to molecule 6 in a solvent like NMP or DMF. Compound 8 can be obtained by reacting compound 7 with DMF and Phosphorus oxychloride under Vilsmeier reaction conditions, Aldehyde 8 can be converted in two steps to substituted ketone 8b by- reacting with a Grignard reagent itiMgX, followed by reaction with an oxidant such as DCC or using Swem reaction conditions,

[00509 J Compound 8 and 8a, or 8b and 8a can react upon heating in a solvent such as ethanol and in the presence of a base such as piperidine to form compound 9. Oxidation of 9 by an oxidant such as meta-chloroperbenzoic acid or oxone can provide compound 10, which can contain variable quantities of sulfide (n = 0), sulfoxide (n = 1) or sulfone (n = 2),

[00510] The chemistry depicted in Figure 5 can be used to prepare various substituted analogs of formula II compounds,

[00511] Compound 10 can be mixed at room temperature or heated with amines R -R_N\1 I to form compound 11. Compound 10 can be reacted with hydrazines R₇¾N-NH₂ to form compound 12, Compound 10 can be reacted with alcohols or phenols R₇OH in the presence of a base such as NaH or ₂CO₃ to form compound 13. Compound 10 can be reacted with thiols or thiophenols R₇SH with or without a base to form compound 14.

[00512] The chemistry described in Figure 6 can be used to prepare analogs substituted by aryl or heteroaryls groups. Compound 7 can be reacted with boronic esters or acids W-B(GR')₂ or organ o tin compounds W-Sn(R')₃ in the presence of tri(2-furyl)phosphine,

copper(I)thiophene-2-carboxylate and Pd₂dba₃ or using conditions previously desbribed in Organic Letters 2002, vol 4(6), pp. 979-981. Compound 15 can be converted to compound 18 using chemistries similar to the one described in Figure 4,

Example 193. Synthesis of 2-fmethylthio )pyrazolo[ 1 ,5-aJ [ 1 ,3,5]triazin-4(3H)-one

[00513] The material was prepared according to a procedure published in patent US

3,846,423. Characterized by LCMS (ES):>95% pure, m/z 183 [ M ! ! | ^' .

Example 194. Synthesis of 4-chloro-2-(methylthio)pyrazolori,5-a][l,3,5]triazinc

[00514] In a round bottom flask equipped with a magnetic stirbar, 2~

(methylthio)pyrazolo[L5-a][l ,3,5]triazin-4(3H)-one (1.0 eq, 10.43 g, 57.24 mmol) was suspended in acetomirile (100 ml), Phosphorus oxychloride (4.0 eq, 21 ml, 229.4 mmol) and triethylamine (1 .05 eq, 8.4 ml, 60.27 mmol) were added and the mixture stirred at reflux for 3.5 hours, at which time LCMS indicated completion of the reaction. The mixture was cooled down and slowly poured into crushed ice (final total volume of about 600 ml). The solid was fi ltered, washed with water and dried in a vacuum oven to afford 4-chioro-2-(methyithio)pyrazolo[l,5- a][l,3,5]triazine as a tan solid (8.15 g, 71 % yield). LCMS (ES):>97% pure, m/z 201 [M+H]⁺.

Example 195. Synthesis of N vck)propvl-2-(nic;hvh io¾pvra/oloi 1 . -a 11 i .3,5 i ria/in-4-aniine

[00515] 4-Chloro-2~(methylthio)pyrazolo[l,5-a][l,3,5]triazine (1.0 eq, 6.26 g, 31 , 19 mmol) was suspended in anhydrous NMP (50 ml). Cyclopropylamine (1 .5 eq, 3.2 ml, 46.26 mmol) was added through syringe dropwise. Internal temperature rose to 47°C, The mixture was stirred without any external cooling for one hour. An additional amount of cypropylamine (1 ml) was added and the mixture stirred for another 1.5 hours. The mixture was slowly poured into water (500 ml) under stirring. The resulting solid was filtered, washed with water and dried in a vacuum oven to give N-cyclopropyl-2-(me1hylthio)pyrazolo[l ,5-a][l ,3,5]triazin-4-amine as a tan solid (5.44 g, 79% yield). LCMS (ES):>95% pure, m/z 222 [M+li .

[00516] The following molecules were prepared using chemistries similar to Example 195 . Compounds were characterized by LCMS,

Table 29.

Example 196, Synthesis of 4-(cyclopropylammo)-2-(methylthio)pyrazolo[ 1 ,5 -a] [ 1 ,3,5]triazine- 8-carbaldehvde

[00517] N-Cyclopropyl-2-(methylthio)pyrazolo[ 1 ,5-a] [ 13,5]triazin-4-amine (1.0 eq, 3.10 g, 14.00 mmol) was dissolved in anhydrous DMF (50 ml ) under nitrogen atmosphere. Phosphorus oxy chloride (5.0 eq, 6.4 ml, 69.9 mmol) was added dropwise over 5 minutes. Internal temperature rose to 45°C, The reaction was stirred in an oil bath at 70°C for 4.5 hours. The mixture was cooled down and added dropwise into a solution of 6N NaOH (150 ml) chilled with an ice bath. The rate of addition was adjusted to maintain the internal temperature of the aqueous NaOH below 16°C, At the end of the addition, the mixture was neutralized by slow addition of 6N HC1 to reach pH = 5-6, The resulting solid was filtered, washed with water and dried in a vacuum oven overnight. 4-(cyclopropylamino)-2-(methyl&^

carbaldehyde was isolated as tan solid (9.26 g, 93%). LCMS (ES):>95% pure, m/z 250 [M+H]⁺. [00518] The following molecules were prepared using chemistries similar to Example 196 . Compounds were characterized by LCMS,

Table 30.

Example 197.

al[l ,3,5]triazin-8-yl methylene)iniidazolidine-2,4-dione

[00519] 4-(Cyclopropylamino)-2-(methylthio)pyrazolo[ 1 ,5-a] [ 1 ,5]triazine-8-carbaldehyde (1 .0 eq, 3.00 g, 12.03 nimol) was suspended in ethanol (40 ml). Hydantoin (1.5 eq, 1.81 g, 18.08 mmol) and piperidine (1 .5 eq, 1 ,78 ml, 18.01 mmol) were added. The mixture was heated at reflux under vigorous magnetic stirring for 3 hours. After cooling of the reaction mixture, the precipitate was filtered, washed with ethanol, then with a mixture of ethanol and water (1 : 1). After drying in vacuo, (Z)-5-((4-(cyclopropy iamino)-2-(methylthio)pyrazolo[ 1 ,5- a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione was isolated as a yellow solid (3.80 g, 95%). LCMS (ES):>85% pure, m/z 332 [M+H] ⁺.

[00520] The following molecules were prepared using chemistries similar to Example 197 . Compounds were characterized by LCMS.

Table 3 1.

Example 198. Synthesis of (Z)- -((4-(cyd

a][l,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z^")-5-((4- (cvclopropylarmno -2-(methylsulfinyl pyrazoloil , 5-a]|^" 1,3,5 ]triazin-8- l)methylene)imidazolidine-2,4-dione

[00521 j (Z)-5 -((4-(Cyclopropylamino)-2-(meth lthio)pyrazolo [ 1.5 -a] [ 1 ,3 , 5 ]triazin-8- yl)methylene)imidazolidine-2,4-dione (1.0 eq, 3.00 g, 9.05 mmol) was suspended in

dichloromethane (150 mi). #j-cpba (77% purity grade, 5.0 eq, 10.1 g, 45.06 mmol) was added and the mixture stirred at room temperature for 4 hours. The reaction was diluted by addition of dichloromethane (500 ml). The solid was filtered and washed with dichloromethane. After drying a (1:1) mixture of (Z)-5-((4-(cyclopropylamiiio)-2-(methylsulfonyl)pyrazolo[ 1 ,5- a][l,3,5]triazm-8-yi)methySene)imidazoiidine-2,4-dione and (Z)~5-((4-(cyclopropylamino)-2- (methylsuiiInyl)pyrazolo[l,5-a][l,3,5]triazin-8-yl)methylene)^ was isolated as a yellow solid (2.67 g, 81 %). LCMS (ES):>85% pure, m/z 364 [M+H]⁺ and m/z 398 [M+H]⁺. The mixture was used for next step without any separation of the molecules.

[00522] The following mixtures of sulfones and sulfoxides were prepared using chemistries similar to Example 198. Compounds were characterized by LCMS.

Table 32.

Example 199. Synthesis of ¾Z)-5-(Y2-f3-ehlorophem

[00523] A (1 : 1) mixture of (Z)-5-((4-(cyclopropylamino)-2-(methy[sulfonyl)pyrazolo[l ,5- a][l,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2- (niethylsulfmyI)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)methylene) (15 mg) was mixed with 3-ciiloroaiiiiiiie (0.1 ml) in NMP (0.2 ml) and the mixture heated in a microwave oven at 1 0 'C for 15 min. Methanol was added and the resulting solid filtered and dried to provide (Z)-5-((2-(3-chlorophenylamino)-4-(cyclopropylamino)pyrazolo[ 1 ,5- a][l ,3,5]triazin-8-yl)methylene)irnidazolidine-2,4~dione as a solid (7 mg). LCMS (ES):>95% pure, m/z 411 [M+H]^÷.

Example 200. Synthesis of (Z)-5-((4-(cyclopropylamino)-2-(cyclopropylmethylamino)-- razolo[L5-aj[l ,3,5]triazin-8-yl) methylene) imidazojidine-2,4-dione

[00524] A (1 : 1 ) mixture of (Z)-5-{(4-(cyclopropylamino)-2-(methylsulfonyi)pyrazolo[1 ,5- a][l,3,5]triazm-8-yl)methylene)irnidazolidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2- (methylsulfinyl)pyrazolo[l,5^ (36 rng) was suspended in NMP (0.2 ml). Cyelopropylmethylamine (88 uL) was added and the mixture stirred at room temperature for 15 minutes, Water and methylene chloride were added and the resulting precipitate was filtered. After triturating in a mixture of ethyl acetate and hexanes, (Z)-5-((4-(cyclopropylamino)-2-^^

methylene) imidazolidine-2,4-dione was isolated as a yellow solid. ). LCMS (ES):>95% pure, rn/z 355 [ M · 1 i i .

Example 201. Synthesis of (Z)-5-((2-(3-chlorophenoxy)-4-(cyclopropy

a jj 1 .3.5 llriazin-8- i )nic;hviene)n)Uiia/oiidinc-2.4-iiic)ii .

[00525] A (1 : 1) mixture of (Z)-5-((4-(cyclopropylamino)-2-(methylsulfoiiyl)pyrazolo[l,5- a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2- (niethylsulfinyl)pyrazolo[l,5- (1 ,0 eq,

25 mg, 0.0704 mraol) was combined in a vial with 3-chlorophenol (5.0 eq, 45 mg, 0.35 mmol) and K₂C0₃ (5,0 eq, 48 mg, 0.347 mmol) in NMP (0.2 ml). The mixture was stirred at 90°C for 1 hour. Water was added and the resulting solid was filtered and dried. Trituration in a mixture of ethyl acetate and hexanes followed by filtration provided (Z)-5-((2-(3-chlorophenoxy)-4- (cyclopropylamino)pyrazolo[l,5-a] as a tan solid (20 mg, 69%). LCMS (ES):>95% pure, m/z 412 | M - ! l i . Example 202, Synthesis of ¾ lr,4r)-4-(4-(cyclopropylamino)-8-((Z)-(2,5-dioxoimidazolidin-4- vlidene methyl)pvrazolori,5-airi,3 ltriazin-2-vlarrijno)-N- methylcyclohexanecarboxamide.

[00526] (lr,4r)-4-(4-(Cyclopropylamino)-8-((Z)-(2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazo[o[l,5-a][l,3,5]triazin-2-ylamino)cyclohexanecarboxylic acid (1 eq, 12 mg, 0,028 mmol) was mixed in NMP (0,4 ml) with methyl amine hydrochloride (8 eq, 15 mg, 0.225 mmol), HOBt.H₂0 (2 eq, 8 mg, 0.056 mmol), DIE A (4 eq, 14 uL, 0.1 13 mmol) and EDO (4 eq, 22 mg, 0.113 mmol). The mixture was stirred at 70°C for 2.5 hours. Water was added and the precipitate filtered to afford (lr,4r)-4-(4-(cyciopropylamino)-8-((Z)-(2,5-dioxoimidazo!idin

4-yUdene)methyl)pyrazolo[l,5-a][1

LCMS (ES):>95% pure, m/z 440 | M I ! j .

Example 203. Synthesis of ( lr,4r)-4-(4-(cyclopropylamino)

ylidene)nieth l)pyrazolo[l,5-a][l,3^

[00527] (lr,4r)-4-(4-(Cyclopropylamino)-8-((Z)-(2,5-dioxoimidazolidin-4- yli dene)methy 1 }pyrazo io [ 1 , 5 -a] [ 1 ,3 ,5 ]triazin-2-ylamino kyclohexanecar boxylic acid ( 1 eq, 12 mg, 0.028 mmol) was mixed in NMP (0.4 ml) with ammonium chloride (8 eq, 12 mg, 0.225 mmol), BOBIH₂O (2 eq, 8 mg, 0.056 mmol), DIE A (4 eq, 14 uL, 0.1 13 mmo!) and EDO (4 eq, 22 mg, 0.113 mmol). The mixture was stirred at 70°C for 2.5 hours, Water was added and the precipitate filtered to afford (lr,4r)-4-(4-(cyclopropylamino)-8-((Z)-(2,5-dioxoirnidazoiidm-4- ylidene)methyl)pyrazolo [ 1„ 5 -a] [1,3,5] triazin-2- lamino)cyclohexa.necarboxamide . LCMS (ES):>95% pure, m/z 426 | M · ! I i .

Example 204. Synthesis of (/.¾-5-({2-{( i r.4r)-4-anr:nocvck; :cxvianr:no)-4-

(c clopropylamino)pyrazolo[l,5-3][l,3,51tri3zin-8-yl)methylene)im

[00528] A (1 : 1) mixture of (Z)-5-((4-(cyclopropylarnino)-2-(methylsulfonyl)py a][1 ,5]triazin-8-yI)niethylene)imi(iazoIidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2- (nietiiy]sulfiiiyljpyrazolo[l ,5-a][l,3,5]tri (1 ,0 eq,

16 mg, 0.0451 mmol) was reacted with ira«s-l ,4~diaminocyclohexane (20.0 eq, 103 mg, 0.902 mmol) in NMP (0.4 ml) at room temperature for 3 hours. Water and methanol was added and the material was purified by preparative HPLC. Genevac evaporation provided (Z)-5-((2-((lr,4r)-4- ammocyclohexylammo)-4-(cyclopropylam

yl)methylene)imidazolidine-2,4-dione 2,2,2-trifiuoroacetate (15 mg). LCMS (ES):>95% pure, m/z 398 j M - ! l i . Example 205, Synthesis of tert-butyl cyclopropyl(2-(3-(trifluoromethoxy)phenyl)pyrazolo[ 1 ,5- a][L3,5jtriazin-4-yl)c3rb3mate

[00529] Di-tert-butyl dicarbonate (327 mg, 1.50 mmol) and DMAP (6 mg, 0.05 mmol) were added to N-cyclopropyl2-(metbylthio)pyrazolo[l,5- ][l,3,5]triazm-4-amine (221 mg, 1 mmol) dissolved in dichloromethane (4 mL). After 1 h, the solution was diluted with EtOAc (100 ml.) and washed successively with water (3 x 100 mL) and brine (100 mL). The organic layer was dried over MgS0 , filtered and concentrated in vacuo to an orange oil. The residue was purified via flash column chromatography (10% EtOAc/hexanes) to afford tert-butyl eyclopropyl(2-(methylthio)pyrazolo[l ,5- ][l,3,5]triazin-4-yl)carbamate (368 mg, 79%). LCMS (ES): >95% pure, m / 322 [M+l]⁺.

Example 206. Synthesis of tert-butyl cyclopropyl(2-(3-(trifluoromethoxy)phmyl)pyrazolo[l_<5- a] [ 1 ,3 ,5 ]triazin-4- l)carbamate

[00530] Note: THF was degassed with a stream ofN₂ for 10 min. in a separate flask, Tert- butyl cyclopropyl(2-(methylthio)p>T-azolo[l ,5-a][ l,3,5]triazin-4-yl)carbamate (100 mg, 0.31 mmol), 3-(triiIuoromethoxy)phenyl boronic acid (1 4 mg, 0.74 mmol), tri(2-furyl)phosphine ί mg, 0.37 mmol), copper(I) thiophene-2-carboxylate (167 mg, 0.88 mmo ), Pd₂dba₃ (24 mg, 0.03 mmoi) were combined. The flask w^ras evacuated and backfilled with N₂. THF (3.7 mL) w^ras added and the reaction was heated to 50 °C for 5 d, The solution was diluted with Et₂0 (40 mL) and washed with 10% H₄OH (3 x 30 mL). The organic layer was dried over MgS0₄, filtered and concentrated in vacuo. The solid residue was triturated with Et₂0 and filtered. The filtrate was concentrated in vacuo and purified via flash column chromatography (2.5-5%

EtOAc/hexanes) to afford tert-butyl cyclopropyl(2-(3-(trifluoromethoxy)phenyl)pyrazolo[ 1 ,5- a][l,3,5]triazin-4-yl)carbamate (116 mg, 85%). LCMS (ES): >95% pure, m/z 436 [M+lf .

Example 207. Synthesis or4 cv lo n) vlan)nu))-2 3-{ ;ri ni n inie h xv ; h nvi jpyra/uioj i .5- al[l,3,5]triazine-8-carbaldeh de

[00531 ] Terf-butyl cyclopropyl(2-(3 -(triiluoromethoxy)pheny i)pyrazolo [ 1 ,5 -a] [ 1 ,3 , 5 Jtriazhi- 4-yl)carbamate was dissolved in dichloromethane (0.7 mL) and trifluoroacetic acid (0.7 mL). After 1 h, the solution was concentrated under a stream of air to give crude 4- (cyclopropylamino)-2-(3-(trifluoromethoxy)phenyl)pyrazolo[l ,5-a][1 ,5]triazme-8- carbaldehyde which was used without further purification. LCMS (ES): >90% pure, m/z 336 [Mrl]⁺. 4-(cyclopropylammo)-2-(3-(trifluorometh^

carbaldehyde (87 mg, 0.26 mmol) was dissolved in DMF (0.8 mL), Phosphorus(V) oxychloride (318 μΐ,, 3.47 mmoi) was added was added dropwise and the reaction was heated to 70 °C. After 6 h, the solution was added dropwise to 6M NaOH (-Ί0 mL) cooled to 0 °C. The pH was adjusted to 7 by the addition of 12N HC1. The precipitate was filtered off and dried in vacuo to furnish 4-(cyclopropylamino)-2-(3-(trit1uoromethoxy)phenyl)pyrazolo[l ,5-a][1

carbaldehyde (71 mg, 75%) as a tan solid. LCMS (ES): >95% pure, m/z 364 [M+l]⁺. Example 208.

(trifluoromethoxy)phenyl^

2,4-dione

[00532] (Z)-5-((4-(cyclopropylamino)-2-(3-(1xifluoromethoxy)phenyl)pyTazolo[l,5- ][l,3,5]triazine-8-yl)methylene)imidazo ine-2,4-dione was prepared using chemistries similar to those exemplified in Example 197. LCMS (ES): >90% pure, m/z 446 [M+lf.

[00533] The following compounds were prepared using chemistries described in Example 1 9, Example 200, Example 201, Example 202, Example 203, Example 204, Example 205, Example 206, Example 207 and Example 208; using appropriate reagents. General methods for preparation of such compounds are included in Figures 3-14 herein. Reagents bearing two reactive amino groups were generally used as mono-Boc protected. The protecting group was removed by reaction with trifluoroacetic acid in methylene chloride prior to purification.

Compounds were isolated by fil tration after addition of water or methanol. Some compounds were purified by preparative HPLC and isolated as TFA salts after evaporation at the Genevac. Compounds were characterized by LCMS. Table 33B shows the biological activities of the compounds listed in Table 33A,

Table 33 A.





IJ43

Table 33B.

CK2: IC50 ΡΪ 2: IC50 AB: MDAMB453 AB: BxPC3

Compound

(μ ) (5 μ,Μ ATP) (μΜ)

T14 <0.1 1.25

U14 <0.1 > 2,5000

V14 <1.0 > 2.5000

W14 <0.1 > 2.5000 6.386 23.384

X14 <0.01 1.1205 0.559 1.517

Y14 <0.01 > 2.5000 3.325 6.748

A15 >1 > 2.5000

B15 <0.01 > 2.5000 2.214 3.394

C15 <1.0 > 2.5000

D15 <0.1 1.5464

El 5 <1.0 > 2.5000

F15 < 1.0 > 2.5000

G15 <0.1 > 2.5000

HI 5 <0.1 > 2.5000

115 <0.1 > 2.5000

J15 <0.01 > 2.5000 1.936 11.99

K15 <0.1 1.6558 5.319 23.688

LI 5 <0.1 > 2.5000

M15 >1 > 2.5000

15 >1 > 2.5000

015 >1 > 2.5000

PI 5 <1.0 > 2.5000

Q15 <1.0 > 2.5000

R15 <1.0 > 2.5000

S15 >1 > 2.5000

T15 < 1.0 > 2.5000

U 15 <1.0 > 2.5000

VI 5 <1.0 > 2.5000

W15 <1.0 > 2.5000

XI 5 <L0 18.336 14.152

Y15 <0.1 > 2.5000 6.603 10.731 CK2: IC50 ΡΪ 2: IC50 AB: MDAMB453 AB: BxPC3

Compound

(μ ) (5 μ,Μ ATP) (μΜ)

Z15 <0.0i > 2.5000 6.283 15.114

A16 <0.1 > 2,5000 17.82 > 30

B16 <0.0i > 2.5000 2.627 5.528

C 16 <0.1 1.7695 1.859 1.841

D16 <0.1 > 2.5000 5.03 17.83

El 6 <0.01 > 2.5000 7.856 11.797

F16 <0.01 > 2.5000 3.545 17.187

G16 <0.01 > 2.5000 19.446

H16 <0.1 > 2.5000 18.952 >30

116 <0.1 > 2.5000 12.508 12.673

J16 <0.1 > 2.5000 1 1.685 18.88

K16 < 1.0 > 2.5000 9.2 12.494

LI 6 <0.1 > 2.5000 9.964 11.216

M16 <0.01 5.059 9.052

N16 <1.0 25.644 > 30

016 <0.1 7.829 > 30

P16 <0.1 10.353 > 30

Q16 <0.1 17.61 1 > 30

R16 <0.1 13.114 19.2

SI 6 <0.01 > 2.5000 4.697 8.049

T16 <0.01 1.9391 2.491 1.712

U16 <0.01 > 2.5000 4.313 8.232

VI 6 <0.01 2.438 5.738 5.492

W16 <0.1 > 2.5000 27.524 9.27

X16 <1.0 > 2.5000 17.619 9.053

Y16 <0.1 > 2.5000

Z16 <0.01 <! .() 14.666 2.909

A17 <0.1 1.8629 12.569 5.872

B17 <0.01 > 2.5000 12.517 6.862

CI 7 <0.1 > 2.5000 11.841 2.389

D17 <0.01 1.884 3.353 8.144 CK2: IC50 ΡΪ 2: IC50 AB: MDAMB453 AB: BxPC3

Compound

(μ ) (5 μ,Μ ATP) (μΜ)

E17 <1.0 > 2.5000

F17 <0.01 > 2,5000 3.97 15.62

G17 <0.1 <1.0

H17 <0.1 <1.0 6.919 6.611

117 <0.01 1.1903 0.442 7.667

J 17 <0.01 > 2.5000 1.416 9.445

K17 <0.1 > 2.5000

1,17 <0.1 2.0781 20.449 5.948

Ml 7 <0.01 > 2.5000 27.014 > 30

N17 <0.1 > 2.5000 13.419 19.135

017 <0.1 > 2.5000 12.459 17.049

P17 <0.1 > 2.5000

Q17 <0.1 1.6281 > 30 2.009

R17 <L0 > 2.5000

SI 7 <0.1 > 2.5000 2.594 6.849

T17 <0.01 <1.0 2.751 4.95

U17 <0.1 > 2.5000 4.554 10.817

V17 <0.1 > 2.5000 13.033 1.55

W17 <0.1 > 2.5000

X17 <0.1 > 2.5000 2.477 5.229

Y17 <0.1 > 2.5000 8.283 3.409

Z17 <0.1 > 2.5000 4.486 9.071

A18 <0.01 > 2.5000 1.478 4.729

B18 <0.01 1.304 2.64 4.027

C 18 <0.01 > 2.5000 1.245 1.388

D18 <0.01 > 2.5000 2.286 4.684

El 8 <0.1 > 2.5000 10.575 9.784

F18 <0.01 > 2.5000 >30 >30

G18 <0.1 > 2.5000 13.472 14.435

HI 8 <0.01 > 2.5000 1.907 4.537

118 <0.1 > 2.5000 8.221 14.522 CK2: IC50 ΡΪ 2: IC50 AB: MDAMB453 AB: BxPC3

Compound

(μ ) (5 μ,Μ ATP) (μΜ)

J18 <0.1 > 2.5000

K18 <0.1 > 2,5000

LI 8 <0.1 > 2.5000

Ml 8 <0.1 > 2.5000

N18 <0.1 > 2.5000 26.829 25.022

018 <0.01 > 2.5000 13.2 > 30

P18 <0.1 > 2.5000 21.799 > 30

Q18 <0.1 > 2.5000 1.368 8.975

R18 <0.1 > 2.5000 8.555 13.435

S18 <0.1 > 2.5000

T18 <0.0 i > 2.5000 3.136 4.168

U18 <0.1 > 2.5000

V18 <0.1 > 2.5000

W18 <0.01 > 2.5000 11.668 > 30

X18 <0.01 > 2.5000 13.548 1 8.585

Y18 <0.1 > 2.5000 18.328 5.013

Z18 <0.01 > 2.5000 11.97 >30

A 19 <0.1 > 2.5000

B19 <0.1 > 2.5000

CI 9 <0.01 > 2.5000

D19 <0.1 > 2.5000

El 9 <0.1 > 2.5000

F19 <0.01 > 2.5000

G19 <0.01 > 2.5000

H19 <0.1 > 2.5000

119 <0.1 > 2.5000

J 19 <0.01 > 2.5000

K19 <0.1 > 2.5000

1,19 <0.1 > 2.5000

M19 <0.01

N19 <0.1 CK2: IC50 ΡΪ 2: IC50 ΑΒ: MDAMB453 ΑΒ: BxPC3

(μ ) (5 μΜ ΑΤΡ) (μΜ) (μΜ)

019 <0.1

P19 <0.01

Q19 <0.01

R19 <0.01

S19 <0.1

Scheme ί

[00534] The compounds described in the following table can be prepared using chemistries described on Figure 7.

Table 34.

[00535] The following molecules can be prepared using chemistries similar to Example Example 207 and Example 208.

Table 35.

[00536] The molecules described in Figure 8 were prepared using chemistries described in Example 201, using bases such as .₂C0₃ or sodium hydride.

OTHERS

[00537] The chemistry described on Figure 9 can be used to prepare analogs of formula 11, 4-bromo-6-chloropyridazin-3 -amine 1 can be reacted with 2 using conditions analogous to the preparation described in the patent application WO2009/100375 to form compound 3.

Compound 3 can react with amine RRRTNH to form compound 4. Compound 4 can be transformed to compound 5 by nucleophiiic substitutions with amines, anilines, alcohols, phenols or thiophenols, in die presence of a base, or by transition metal catalyzed conversions such as Suzuki coupling with boronic acid or esters of formula WB(OR.)₂. Compound 5 can be transformed to compound 6 by reduction with LiAlH₄. Alcohol 6 can be converted to aldehyde 7 by oxidation with DCC or under Swera conditions, Compound 5 can react with an

organometa!lic reagent exemplified by Grignard reagent R⁴MgX to form secondary alcohol 8. This compound can be converted to alkyiketone 9 under conditions analogous to the conditions used to convext 6 into 7. Compounds 7 and 9 can both be converted to compound 11 by condensation with 10 in a solvent such as ethanol and in the presence of a base such as piperidine.

[00538] The compounds described in the following table can be prepared using chemistry described on Figure 9.

[00539] Figures 10-14 illustrate other synthesis methods that can be used to prepare compounds of the invention.

Table 36.

Example 209. Synthesis of 5-((5-(3-chlorophenylamino)-7-(cvclopropylamino^')pyrazol.o[T ,5- a] pyrimidin-3 ~ l)methy 1 e e)imidazolidine-2 ,4-di one

[00540 J Step A. To 5,7-diehloropyrazolo[i,5~a]pyrimidine (200 mg, 1.06 mmol) in acetonitrile was added Et₃N (148 μΐ, 1.06 mmol) and cyclopropylamine (75 μΐ, 1.06 mmol). The reaction was heated at 80 °C overnight. The mixture was concentrated under reduced pressure, dissolved in dichloromethane, and washed with water. The resulting organic layer was dried over Na₂S0₄ and concentrated under reduced pressure to afford 156 mg of 5-chloro-N- cyclopropylpyrazolo -a]pyrimidin-7-aniine (70% yield). LCMS (M+ 1=209)

[00541] Step B. To 5-chloro-N-cyclopropylpyrazolo[l ,5-a]pyrimidin~7-amine (156 mg, 0.75 mmol) in DMF was added POCI₃ (205 μΐ, 2.25 mmol). The mixture was stirred at room temperature for 3 hours, Ice was added to quench POCI₃, and then the mixture was neutralized with 1 M NaOH. Dichloromethane was added and the product was extracted three times. The organic layer was dried over Na₂S0₄ and concentrated under reduced pressure to yield 5-chloro- 7-(cyclopropyiamino)pyrazolo[l,5-a]pyrimidme-3-carbaldehyde. Some residual DMF could not be removed. I .CM · 1 237 }

[00542] Step C. To 5-chloro~7~(cyclopropylamino)pyrazolo[l ,5-a]pyrirnidine-3~

earbaldehyde (177 mg, 0,75 mmol) in 1 ,4-dioxane was added 3-chloroaniline (397 μΐ,

3.75 mmol). The mixture was heated in microwave at 120°C for 60 minutes. Precipitate was filtered off, and the filtrate was prepared by TLC (1% methanoi/^'dichloromethane) to yield 26 mg (1 1% yield) of 5-(3-chloropheny3.amino)-7-(cyclopropylamino)pyrazolo[l,5- a]pyrimidine~3~c =328)

[00543] Step D. To 5-(3-chlorophenylamino)-7-(cyclopropylamino)pyrazoio[l,5- a]pyrimidine-3 -earbaldehyde (26 mg, 0.08 mmol) in EtOH was added hydantoin (8 mg, 0.08 mmol) and piperidine (8 μΐ, 0.08 mmol). The mixture was stirred at 70°C for 3 days.

Insolubles were filtered off, and iiltrate was concentrated under reduced pressure. Filtrate was then dissolved in methanol and purified by HPLC to yield 5-((5-(3-chlorophenylamino)-7-

(cyclopropyiammo)pyrazolo[! ,5-a]pyr LCMS (M+l =410) Example 210. Synthesis of 5-^

a]pyrimidin-3- l)methylene)thiazolidine-2,4-dione

[00544] Step A. To 5-chloro-7-(cyclopropylamino)pyrazolo[l,5-a]pyrimidine-3- earbaldehyde (440 mg, 1.86 mmol) in EtOH was added thiazolidine-2,4-dione (458 mg, 3.91 mmol) and piperidine (208 μΐ, 2.05 mmol). The reaction was heated at 80°C overnight, 3 mL Isopropanol was added in the morning, along with 218 mg thiazolidine-2,4-dione, 94 uL piperidine. Temperature was increased to 90°C and left overnight. Precipitate was filtered while hot and dissolved in methanol. 1 mL of 1M HC1 was added and the mixture sonicated.

Precipitate was filtered and washed with methanol to yield 340 mg (54% yield)

5-((5-chloro-7-(cyclopropylaniino)pyrazoio[l ,5-a]pyri

dione as a yell =336)

[00545] Step B. To 5-((5-chloro-7-(cyclopropylamino)pyrazolo[l,5-a]pyrimidin-3- yl)methylene)thiazolidine-2,4-dione (30 mg, 0,09 mmol) in -methylpyrrolidinone (NMP) was added 2-methylpropan-l -amine (20 mg, 0.268 mmol). The reaction was heated at 130°C overnight. Mixture was diluted with methanol and prepared by HPLC to yield 5-((7- (cy cl opropylamino)-5 -(i sob uiylamino)pyrazol o [ 1 ,5 -a] pyrimidm-3-yl)methyiene)thiazoiidme- 2,4-dione. LCMS (M+l =373)

Example 21 1. Synthesis of 5-ff 7-( cyclopropylamino)-5-¾2-hydroxypropylamino )pyrazolo 1 ,5

[00546] The titled compound was prepared using a method analogous to that described Example 21 0, LCMS (M+l=375)

Example 212.

a]pyrimidin-3- l)methylene)thiazolidine-2,4-dione

[00547] The titled compound was prepared using a method analogous to that described for Example 210, LCMS (M+l=373) Example 213. Synthesis of 5-((7-(cyclopro^^

a]pyrimidin-3- l)methylene)thiazolidine-2,4-dione

[00548] The titled compound was prepared using a method analogous to tha described for Example 210. LCMS (M+l=345)

Example 214, Synthesis of 5-((7-(cyclopropylamino -5-(methyl(l-methylpyrrolidin-3- yl)armno yrazolof l,5~a]pyrim

[00549] The titled compound was prepared using a method analogous to that described Example 210, LCMS iM ! 4 14)

Example 215. Synthesis of 5-^

a]pyrimidin-3- l)methylene)thiazolidine-2,4-dione

[00550] The titled compound was prepared using a method analogous to that described for Example 210, LCMS (M+l=363)

Example 216. Synthesis of 5 -((7-(cyclopropylamino)-5 -(4-methy 1- 1 ,4 -diazepan- 1 -

[00551] The titled compound was prepared using a method analogous to that described Example 210. LCMS iM ! 4 14)

Example 217.

pyrazolo l,5-a]pyrimidin-3-yl)methylene)thiazolidine-2,4-dione

[00552] The titled compound was prepared using a method analogous to that described for Example 210. LCMS (Mr 1=416)

Example 21 8. Syr; thesis of 5 ~((5 -(3 -chl orophenylarmno)-7-(cy clopropyj amino)pyrazoio [ 1 ,5 - a pyrimidin-3- Dmethylene)thiazolidine-2,4-dione

1 0553 J To 5-((5-chloro-7-(cyclopropylamino)pyrazolo[ 1 ,5-a]pyrimidin-3- yl)methylene)thiazolidine-2,4~dione (20 mg. 0.06 mmol) in NMP was added 3-chloroaniline (38 μΐ.,, 0.36 mmol) and few granules p-toluenesulfonic acid. The reaction was heated in microwave at 180°C for 1.5 hours. Mixture was filtered and prepared by HPLC then preparative TLC (1% methanol/dichloromethane) to yield 5-((5-(3~chlorophenylamino)-7~

(cyclopropylamino)pyrazolo[l ,5-a]pyrimidm-3-yl)meth.ylene)thiazolidme-2,4-dione as a yellow solid. LCMS (Mr 1-427) Example 219. Synthesis of 5-((7- cycl.opropylamino)-2,^

yr)methylene^")thiazolidine-2,4-dione

[00554] Step A. 7-(Cyclopropylamino)-2,5-diraethylpyrazolo[l ,5-a]pyrimidine-3- carbaidehyde was prepared from N-cyclopropyl-2,5-dimethylpyrazolo[ 1 ,5-a]pyrimidin~7-a.mine using methods analogous to those described in Example 209, Step B, LC S (M+l^::::231)

[00555] Step B. To 7-(cyclopropylamino)-2,5-dimethylpyrazolo[ 1 ,5-a]pyrimidine-3- carbaldehyde (0.25 mmol) in DMF was added thiazolidine-2,4-dione (88 mg, 0.75 mmol) and piperidiiie (25 μΐ, 0.25 mmol). The mixture was stirred at room temperature overnight. Mixture was prepared by HPLC to yield 5-((7-(cyciopropylamino)-2,5-dimethylpyrazoio[ Ϊ ,5- a]pyrimidin-3-yl)methylene)thiazolidme-2,4-dione. LCMS (M+ 1=330) Example 220.

yl methylene thiazolidme-2,4-dione

[00556] Step A, 7-(CyciopropylamuK^)pyrazolo[l,5~a]pyrimidine-3-carbalde]^ was prepared from N-cyclopropylpyrazo!o[l ,5-a]pyrimidm-7-amme using the methods described in Example 219, Step =203)

[00557] Step B. The titled compound was prepared from 7-(cyclopropylamino)pyrazolo[l,5- a]pyrimidine-3-carbaldehyde using methods analogous to those described in Example 219, Step B, except the product was isolated by filtration, washed with methanol, and air dried. LCMS ( M ! 302)

Example 221. Synthesis of 5-^

yl pyTazolo[l,5-a rimidin-3-yl)methylene)imidazolidine-2,4-dione

[00558] Step A. To 5-chloro-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidine-3- carbaldehyde (400 mg, 1.70 mmol) in EtOH was added hydantoin (186mg, 1 ,86mmol) and pyrrolidine (14₍uL, 0.17mmol). The reaction was stirred at 70°C over weekend. Precipitate was filtered and air dried to yield 180mg (33% yield) 5-((5-chloro-7-(cyclopropylamino)pyrazolo- [l,5-a]pyrimid -3-yl)methyiene)imidazolidine-2,4-dione. LCMS ( M 1 1 ;·

[00559] Step B. To 5-((5-chloro-7-(cyclopropylamino)pyrazolo[l,5-a]pyrimidin-3- yl)methylene)imidazoHdine-2,4-dione (30 mg, 0.09 mmol) in 1 ,4-dioxane was added l-(pyridin- 2-yi)piperazine (58 μί,, 4.10 mmol) and Et_¾N (13 L, 0.09 mmol). Reaction was then heated 120°C for 35 minutes in microwave. Solvent was removed under reduced pressure, and mixture was dissolved in methanol. Solid was isolated by filtration, then air dried to yield 11 mg (26% yield) 5-((7-(cyclopropylammo)-5-(4-(pyridin-2-yl)piperazm-l-yl)pyrazolo[l,5-a]pyrimidin-3- yl)methylene)iniidazolidine-2,4-dione. LCMS (M+ 1 =446) Example 222.

a]pyrimidin-3- l)methylene)imidazolidine-2,4-dione

[00560 J The titled compound was prepared using methods analogous to those described for Example 221 , Step B, with the following alteration. The solvent was removed under reduced pressure, and the mixture was dissolved in methanol. The mixture was filtered, and the filtrate was concentrated under reduced pressure to provide 18 mg (48% yield) of the product as a yellow solid. LCMS (M+ 1=397)

Example 223. Synthesis of fZVS-r^-fcyc opropylamino^-O.^- dimethylbenzyiammo^yrazolofl ,5-a][l ,5]triazin-8-yl )methylene)imidazolidine-2,4- dione.

[00561] A solution of (3 ,4-dirn.ethylphenyl)methanamine in NMP (106 μΐ, 0,4 , 1 ,5 eq, 0,042 mmol) was transferred in a glass reaction vial. A solution of a (1 : 1) mixture of (Z)~5-((4- (cyclopropylamino)-2-(methy lsuifonyl)pyrazoio[ 1 ,5-a] [ 1 ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2- (methylsulfinyl)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione in NMP (ΙΟΟμΙ, 0.282M, 1.0 eq, 0,0282 mmol) was added, The mixture was heated at 80 °C for 5 hours. NMP was added (0.7 ml) and the solution subjected to preparative HPLC purification. Genevac evaporation provided (Z)-5-((4-(cyclopropylamino)-2-(3,4-dimethylbenzylamino)pyrazolo[l,5- a][1 ,51triazm-8-yi)niethylene)imidazoiidine-2,4-dione as a solid (5,8 mg). LCMS (E8):>85% pure, m/z 419 | · ! 1 | .

Example 224. Synthesis of (Z)-5-((4-(cyclopropylamino)-2-(l-(pyridin-2- yl)ethylarrnno)pyrazolo[l,5-a][l,3^

[00562] A (2: 1) mixture of (Z)-5-((4-(cyclopropylamino)-2-(methy[sulfonyl)pyrazolo[l ,5- a][l,3₅5jtriazin-8-yl)methylene)irn.ida olidine-2,4-dione and (Z)-5-((4-(cyclopropylarmno)-2- (methylsulfinyl)pyrazolo[l ,5-a][l,3^ (1.0 eq,

3.6 g, 10.08 mmol) was suspended in 2-propanol (40 ml). Rac- 1 -pyridmil-2-yl-eth.ylamme (2,0 eq, 2.47 g, 20.22 mmol) was added and the mixture stirred at 90 °C for 6.5 hours. The mixture was cooled down and the solid isolated by filtration. After drying in a vacuum oven, (Z)-5-((4- (cyclopropylamino)-2-(l-(pyridin-2-yl)ethylamino)pyrazolo[l ₅5-a][l ,3,5]triazi^·n-8- yl)methylene)imidazolidine-2,4-dione was isolated as a pale yellow solid (3.60 g, 88%). LCMS (ES):>95% pure, m/z 406 [M+Ii .

[00563] The compounds in the following table were prepared using procedures described in Example 223, Example 224, Example 199 and Example 200. When the amine reagent was used as a salt, a stoichiometric amount of DIEA was added to the reaction mixture. Table 37B shows the biological activities of the compounds listed in Table 37A.

Table 37A.

300

Table 37Β.

Example 225. Synthesis of (Z)-5 -(( 4-( cyclopropy 1 amino)-2-( 1 H-imidazol- 1 -yl)pyrazolo [1,5

.zin-8-vi)methylene)imidazoiidme-2,4-dione.

[00564] A mixture of (Z)-5-((4-(cyclopropylamino)-2-(methyls^'ul^'finyl)pyTaz;olo[ 1 ,5- a][l ,3,5]triazin-8-yl)niethyiene)imidazolidme-2,4-dione and (Z)-5-((4-( cycloprop lamino)-2- (methylsulfonyl)pyrazolo[l,5-a]^^ (10 nig,

0.028 mmol) was mixed with imidazole (6 mg, 0.084 mmol) in isopropanoi (I ml.,). The mixture was stirred at 80 °C for 16h. The reaction mixture was cooled to room temperature and the resulting solid was filtered off and washed with isopropanoi . The solid was dried under vacuum to provide (Z)-5-((4-(cyclopropy[amino)-2-(lH-imidazol-l-yl)pyrazolo[ l,5-a][l ,3,5]triazin-8- yi)methylene)imidazoiidine-2,4-dione as a yellow solid. LCMS (ES):>95% pure, m/z 352

[M-H-if. Example 226. Synthesis of (Z)-l -(4-(cy

ylidene )methyl)pyrazolo [ 1 ,5 -a] [ 1 ,3 ,5 Jtriazin-2-y 1 ^")- 1 H-benzo [d]imidazole-5 -earboxylic

acid.

[00565] A mixture of (Z)-5-((4-(cyclopropylamitio)-2-(methylsulfinyl)pyrazolo[ 1 ,5- a][l ,3,5]triazm-8-yl)methylene)irnidazolidine-2,4-dione and (Z)-5-((4-(cycloprQpylarrmio)-2- (methylsulfonyl)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)methylene)irriidazolidine-2,4-dione ( 10 nig, 0.028 mmol) was mixed with 1 H-benzo [d]imidazole-5 -carboxylic acid (20 mg, 0.140 mmol) in isopropanol (I niL). The mixture was stirred under microwave heating at 150°C for 20 minutes. The solvent was removed to provide (Z)-l-(4-(cyc[opropylamino)-8-((2,5-dioxoimidazolidin-4- yiidene)meihyi)pyrazolo[l ,5-a][l ,3,5]triazin-2-yl)-l H-benzo[d]imidazole-5-carboxylic acid as a crude mixture which was taken on to the next step without further purification. LCMS

(ES):>95% pure, m/z 446 \ M I I I ^'

Example 227. Synthesis of (Z)-5-((4-(cyciopropyja.mmo)-2-( ^'5-(4-ethyl.piperazme- 1 -carbonyl)- lH-benzo[d]imidazol-l-yl)pyrazolo[L5-a][l ,3,5]triazin-8-yi)methylene)™^

dione.

[00566] To (Z^-l-(4-(cyclopropylamino)-8-((2,5-dioxoirn.idazolidin-4- yli dene)methy l)pyrazo io [ 1 , 5 -a] [ 1 ,3 ,5 ]triazin-2-yl)- 1 H-benzo [d] imidazole-5 -carboxylic acid ( 15 mg, 0,034 mmol) in DMF (2 in! . } was added EDCI (65 mg, 0.34 mmol), HOBt (46 mg, 0,34 mmol), and 1-ethyipiperazine (44 μΕ, 0.34 mmol). The mixture was stirred at 50 °C for Ih. The reaction mixture was cooled to room temperature, filtered through PTFE filter, and purified by mass-directed LC/'MS to provide (Z)-5-((4-(cyclopropylamino)-2-(5-(4-ethylpiperazine- 1 - carbonyl)-lH-benzo[d]imidazol-l -yl)pyrazolo[l ,5-a][l ,3,5]triazin-8- yl)methylene)imidazolidine-2,4-dione as the TFA salt, LCMS (ES):>95% pure, m/z 542

| M 1 1 ] .

Example 228. Synthesis of (Z)-l-(4-(cyclopropylammo)-8-((2,5-dioxoimidazolidin-4- -aj[ 1 ,3.5jiria/in-2-yi )- 1 i i~imld /oic~4~cnrbnidchydc.

[00567] A mixture of (Z)-5-((4-(cyclopropylammo)-2-(methyls^'ul^'finyl)pyTaz;olo[ 1 ,5- a][l,3,5]triazm-8-yl)methylene)imidazolidine-2,4-dione and (Z)-5^(4-(cyclopropylamino)-2- (methylsuifonyi)pyrazoio[l ,5-a][l ,3,5]triazin-8-yl)rnethyiene)imidazolidine~2,4-dione (20 mg, 0.056 mmol) was mixed with lH-imidazole-4-carbaldehyde (16 mg, 0,168 mmol) in isopropano (2 fflL). The mixture was stirred under microwave heating at 150 °C for 20 minutes. The reaction mixture was cooled to room temperature and the solvent was removed by rotary evaporation to provide (Z)-l-(4-(cyclopropylamino)-8-((2_!l5-dioxoimidazolidm-4- ylidene)methyl)pyrazoio[l ,5-a][l ,3,5]triazin-2-y[)-l H-imidazole-4-carbaldehyde as a crude mixture which was taken on to the next step without further purification. LCMS (ES):>95% pure, m/z 380 I M l ! j ^' Example 229, Synthesis o^^

imidazole -yl)pyrazolo[l,5-a] l 3, 5]tm

[005681 To (Z)-l -(4-(cyclopropyiamino)-8-((2,5-dioxoimidazoiidm-4- ylidene)methyl)pyrazolo[l,5-a][l ,3,5]triazin-2-yl)-lH-imidazole-4-carbaldehyde (7 mg, 0.018 mmol) in DCE (1 mL) was added pyrrolidme (10 mg, 0.144 mmol) and sodium

triacetoxyborohydride (36 mg, 0, 144 mmol). The reaction mixture was stirred under microwave heating at 120 °C for 10 minutes, Dilute with DMSO (1 mL) and filter through PTFE filter. Purified by mass-directed LC/MS to provide (Z)-5-((4-(cyclopropylamino)-2-(4-(pyrrolidin-l- ylmethyl)- 1 H-imidazol- 1 -yl)pyrazolo[ 1 ,5-a] [1,3,5 ]triaziii-8-yi)metiiylene)imidazoiidine-2,4- dione as the TFA salt. LCMS (ES):>95% pure, m/z 435 [M+H]+

[00569] The compounds in the following table were prepared using chemistries described in Examples 225 to 229. Table 3813 shows the biological activities of the compounds listed in Table 38 A.

Table 38A.

Table 38B.

Example 230. Synthesis of (Z -5-(^"(2-(3-chloroph.enoxy^')-4-fcvcl.opropylamino)pyrazolon ,5- -8-yl)methylene)imidazolidine-2,4-dione.

[00570] A (1 : 1) mixture of (Z)-5-((4-(cyclopropyiarnino)-2-(methyisulfonyl)py a][1 ,5]triazin-8-yI)niethylene)imi(iazoIidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2- (niethy]suifiiiyl)pyrazolo[l ,5-a][l,3,5]tri (1 ,0 eq,

25 mg, 0.0704 mmo!) was combined in a vial with 3-chlorophenol (5.0 eq, 45 mg, 0.35 mmol) and K₂CO₃ (5.0 eq, 48 mg, 0.347 mmol) ui NMP (0.2 ml). The mixture was stirred at 90 °C for 1 hour. Water was added and the resulting solid was filtered and dried. Trituration in a mixture of ethyl acetate and hexanes followed by filtration provided (Z)-5-((2-(3-chlorophenoxy)-4- (cyclopropylamino)pyrazolo[l ,5-a][l ,3,5]triazin-8-yl)niethylene)i as a tan solid (20 mg, 69%). LCMS (ES):>95% pure, m/z 412 | Μ · ! 1 ; .

[00571] The following compounds were prepared using the chemistry described in Example 230. Table 39B shows the biological activities of the compounds listed in Table 39A.

Table 39A.

322

323

Table 39B.

Example 231. Synthesis of (Z)-5-((2-(bcnzyloxy)-4-(cyclopropylamino)pyrazolo[ 1 ,5- -8-yl)metliylene)imidazolidine-2.4-dione.

[00572] Benzyl alcohol (14.2 μΐ,, 0.138 mmol) was dissolved i NMP (0.2 mi). Sodium Hydride (60%, 5.5 mg, 0, 138 mmol) was added and the reaction stirred at room temperature for one hour. A ( 1 : 1) mixture of (Z)-5-((4-(cyelopropy!amino)-2-(niethyisulfonyl)pyrazolo[l,5- a][l,3,5]triazin-8-yi)methylene)imidazoiidine-2,4-dione and (Z)-5-((4-(cyclopropylamino)-2- (methylsulfinyl)pyrazolo[l,5-a][l,3,5jtriazin-8-yl)methylene)im azolidine-2,4-dione (10 mg, 0,027 mmol) was added and the mixture was stirred at room temperature for one hour, Water was added and the material was extracted with ethyl acetate. After concentration at the rotary evaporator, addition of methanol formed a precipitate that was filtered and dried, (Z)-5-((2- (benzyloxy)-4~(cyclopropyiam

2,4-dione was isolated as a solid (5.6 mg). LCMS (ES):>95% pure, m/z 392 [M+H]+. [00573] The following compounds were prepared using conditions similar to Example 231. Table 40B shows the biological activities of the compounds listed in Table 40A,

Table 40A.

Table 40B.

Example 232. Synthesis of 3-{( lr,4r -4-(7-(cyclopropylamino)-3-((Z)-{2.5-dioxoimidazolidin- 4-ylidene)methyl)pyrazolo[ 1 ,5-a]pyrimidin-5-ylamino)cyclohexyl)- 1 , 1 -dimethylurea 2,2,2- trifluoroacetate

[00574] (Z)-5 -((5 -(( 1 r,4r)-4-aminocyclohexylamino)-7-(cycl opropylamino)pyrazolo [ 1,5- a]pyrimidin-3-yl)methykine)irnidazolidine-2,4~dione 2,2,2-trifluoroacetate (10 mg) and DIEA (1.2 eq, 4,1 ul) were mixed in dry NMP (0.1 ml), Dimethylcarbamic chloride (1.0 eq, 1.8 ul) was added and the mixture stirred at room temperature overnight. The reaction was diluted with NMP (1 .5 mi) and a few drops of water. The compound was purified by preparative HPLC and was isolated after evaporation at the genevac, 3-((lr,4r)-4~(7-(cyclopiOpyiammo)-3-((Z)-(2,5- dioxoirmdazolidin-4-ylidene)niethy])pyrazolo[ 1 ,5-a]pyrimidin-5-ylamino)cyclohexyl)- 1,1- dimethylurea 2,2,2-trifluoroacetate. LCMS (ES):>95% pure, m/z 468 [M+H]+. Z:E ratio: 86: 13.

Example 233. Synthesis of -(( lr,4r)-4-(7-{cyclopropylamino)-3-({Z)-(2,5-dio

-ylidene)methyl)pyrazolo[ 1 ,5-a]pyrimidin-5-ylainino)cyclohexyl)acetamide

[00575] (Z)-5 -((5 -(( 1 r,4r)-4-ammocycl ohexy 1 amino)-7-(cyclopropy ί ammo)pyrazolo [1 ,5- a]pyrimidin-3-yl)metliylene)imidazolidine-2,4-dione 2,2,2-trifiuoroacetate (1.0 eq, 10 mg, 0.0196 mmol) and DIEA (1.2 eq, 4 ul, 0.0229 mmol) were dissolved in NMP (0, 1 mi). Acetic anhydride (1.0 eq, 2 ui, 0.0211 mmol) was added and the mixture stirred at room temperature overnight. Water was added and the resulting precipitate was filtered and dried to provide N- ((lr,4r)-4-(7-(cyc!opropyiamino)-3-((Z^

a]pyrimidin-5-ylamino)cyclohexyl)aceta.mide as a solid (8 mg). LCMS (ES):>95% pure, m/z 439 | M i l ] .

Example 234. Synthesis of -((lr,4r)-4-(7-(cyclopro

4-ylidene)methyl)pyrazolo j^" 1 ,5 -a]pyrimidin-5 -ylamino)cyclohexyl)propionamide 2 ,2,2- trifluoroacetate.

1 0576J Z)-5 -( { 5 -(( 1 r,4r)-4-aminocyc 1 ohexylamino)-7-(cyciopropylamino)pyrazoio [1 ,5- a]pyrimidiii-3-yi)methylene)imidazoiidine-2,4-dione 2,2,2-trifluoroacetate (1.0 eq, 10 mg, 0.0195 mmol) was suspended in NMP (0.05 ml). A NMP solution of propionic acid (1.2 eq, uL of 0.4 M solution, 0.0234 mmol), HOBt (1.5 eq, 4 mg, 0.030 mmol), DIEA (2.5 eq, 8 uL, 0,048 mmol) and EDCI (1.5 eq, 6 mg, 0.03 mmol) were added and the mixture stirred at 70 °C for 1.5 hours. Water and NMP was added and the product was purified by preparative HPLC, Genevac evaporation provided N-((lr,4r)-4-(7-(cyciopropylamino)-3-((Z)-(2,5- dioxoimidazolidin-4-ylidene)niemyl)pyrazolo[ 1 ,5-a.]pyrimidin-5- ylamino)cyclohexyl)propionamide 2,2,2-trifluoroacetate (2.8 mg). LCMS (ES):>90% pure, m/z 453 J · ! ! J · .

Example 235. Synthesis of (Z^")-5-((4-(cyclopropylamino -2-((lr,4r)-4- (isobutylamino)cyclohe^

yl)methylene)imidazolidine-2,4-dione 2,2,2-trifluoroacetate.

[00577] (Z)-5-((2-((lr,4r)-4-armnocyclohexylarmno)-4-(cyclopropylamino)pyrazolo[l,5- a][l ,3,5]triazin-8-yl)methylene)imidazolidine-2,4-dione 2,2,2-trifluoroacetate (1.0 eq, 10 mg, 0.025 mmol) was suspended in dichloroethane, Isobutyraldehyde (4 eq, 9.2 uL, 0.101 mmol), DIEA (! .0 eq, 0.025 mmol) and NaBH(OAc)₃ (4 eq, 21 mg, 0.101 mmol) were added and the mixture was stirred at room temperature for two hours. The mixture was diluted with water and NMP and subjected to preparative HPLC purification. Genevac evaporation provided (Z)-5-((4- (cyclopropylaniino)-2-((lr,4r)-4-(isobuty[amino)cyclohexylamino)pyrazolo[l,5-a][l ,3,5]triazin- 8-yi)niethylene)imidazolidine-2,4-dione 2,2,2-trifluoroacetate as a solid (4,6 mg). LCMS (ES):>90% pure, m/z 454 j M · 11 i · .

[00578] The compounds in the following table were prepared using procedures described in Examples 232 to 235 and Examples 30 and 31 , using the appropriate starting amines and carboxylic acids, acyl chlorides, sulfamoyl chlorides, sulfonyl chlorides, isocyanates and chloro formates. Table 41 B shows the biological activities of the compounds listed in Table

41 A. Table 41 A.

335

Table 4 IB.

Example 236. Synthesis of (Z)~5-((5~(3~chloTO

-3-yl)methylene)imidazolidine-2,4-dione.

[00579] Tert-bntyl 5-chloro-3-forrnylpyrazolo[1 ,5-a]pyrimidin-7-ylfeyclopropyl)carbamate (1 ,0 eq, 49 nig, 0.145 mmol) was mixed in a vial with NMP (0.2 mi), 3-chiorophenol (5.0 eq, 93 mg, 0.274 mmol) and potassium carbonate (5.0 eq, 100 mg, 0.723 mmol). The reaction mixture was stirred at room temperature for one hour. Water was added and the resulting gummy material was extracted with methylene chloride. The organic phase was dried over Na₂S0₄ and the volatiles removed in vacuo. The resulting NMP solution was reacted with a HCI 4N solution in dioxane (5 mi) at room temperature for one hour, at which time LC 8 monitoring indicated completion of the reaction. The reaction was treated with water and 6 NaOH and stirred overnight at room temperature. The solid was filtered and dried to afford crude 5-(3- ch!orophenoxy)-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidine-3-carbaidehyde as a solid (31 mg). The material was heated with hydantoin (30 mg), piperidine (30 uL) in Ethanoi (1 ml) in a vial at 90 °C for seven hours. Water was added and the materia! was filtered, washed with ethanoi, ethanoi/ water and dried. (Z)-5-((5-(3-dilorophenoxy)-7-

(cyc!opropylamino)pyrazolo[l,5-a]pyrimidm-3-yi)methy!ene)imidazoiidine-^ was isolated as solid (43 mg, 38% over 2 steps). ). LCMS (ES):>95% pure, m/z 41 1 [M+H]⁺.

[00580] The following compounds were prepared using chemistry described in Example 236. Table 42B shows the biological activities of the compounds listed in Table 42A. Table 2Λ.

[00581] 2-cbloro-3-mtropheno3. (1 g, 5.8 mmol) was dissolved in D F (6 mL). K. ;( ( ) ; (1.6 g, 11.5 mmol) was added and the solution changed from yellow to red. 4~(2- chloroethyl)morpholine hydrochloride (1.07 g, 5.8 mmol) was added and the solution was allowed to stir for 15 h. The reaction was poured into H₂0 (30 mL) and extracted with EtO Ac (3 x 30 mL). The orgamcs were washed with IN NaOH (100 mL) and brine (100 rnL) and then dried over MgS0₄, filtered, and concentrated in vacuo to provide 4-(2-(2-chloro-3- nitrophenoxy)ethyl)morphoime (1.4 g, 87%) as a golden oil.

[00582] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 237. All compounds were characterized by LCMS.

Example 238. Synthesis of 2-chloro-3-{2-morpholinoethoxy)anil in e

[00583] 4-(2-(2-chloro-3-mtrophenoxy)ethyl)morpholine (873 nig, 3,1 mmol) was dissolved in toluene (12 mL). Ammonium formate (866 mg, 13.7 mmol) was dissolved in H₂0 (12 mL) and added. Iron powder (<10 micron, 766 mg, 13.7 mmol) was added and the reaction was placed in a 120 °C oil bath. After 1.25 h, the solution was cooled to 23 °C and filtered over a pad of celite eluting with 10% MeOH/dichloromethane (250 mL). The filtrate was concentrated in vacuo and the residue was purified via flash column chromatography to give 2~chloro~3-(2- morpholinoethoxy)aniime (503 mg, 75%) as a golden oil.

[00584] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 238. All compounds were characterized by LCMS.

Example 239. Synthesis of 2-ehloro-4-( -pyrazol- 1 -ypanilme

[00585] 2-chloro-4-iodoaniline (760 mg, 3 mmol), N, /V-dimethyiethylenediamine (96 μΐ., 0.9 mmol), IH-pyrazole (430 mg, 6.3 mmol) were dissolved in DMF (3.8 mL). ί s X> _; (1.86 5.7 mmol) and Cul (57 mg, 0.3 mmol) were added and the reaction was placed in a 140 °C oil bath. After 3 h, the volatiles were removed in vacuo. The residue was diluted with

dichlorom ethane and purified via flash column chromatography (1 % MeOH/dichloromethane) to furnish 2-chloro-4-(lH-pyrazol-l-yl)amline (540 mg, 93%) as a golden brown oil which crystallized overnight at -20 °C. [00586] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 239. All compounds were characterized by LCMS.

Example 240. Synthesis of 1 -(3-chloro-4-nitrophenyl)-4-methylpiperazine

[00587] 2-chloro-4~fiuoronitrobenzene (1 g, 5.7 mmoi) was dissolved in DMF (10 mL). 1- methylpiperazvne (760 μΐ,, 6.8 mmol) and then ₂C0₃ (1 .57 g, 1 1.4 mmoi) were added and the reaction was placed in a 100 °C oil bath. After 1 h, the solution was cooled to 23 °C, then added to ¾0 (75 mL). The precipitate was filtered, washed with H₂0 (-25 mL), then dried overnight (50 °C, 25 mmHg) to provide l -3-chloro-4-nitrophenyl)-4-methylpiperazine ( 1.28 g, 88%) as a maize colored solid.

[00588] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 240. All compounds were characterized by LCMS.

Exam le 241. Synthesis of 2-chloro-4-(4-methylpiperazin- l-yl)aniline

[00589] l -3-chloro-4-nitropheny[)-4-methylpiperazine (414 mg, 1.62 mmol)was dissolved in toluene (6,5 mL), Ammonium formate (461 mg, 7.3 mmol) was dissolved in H₂O (6,5 mL) and added. Iron powder (<10 micron, 408 mg, 7.3 mmol) was added and the reaction was placed in a 120 °C oil bath. After 1.25 h, the solution was cooled to 23 °C and filtered over a pad of celite eluting with 10% MeOH/dichloromethane (250 mL). The filtrate was concentrated in vacuo and the residue was partitioned between H?0 (25 mL) and EtOAc (25 mL), The aqueous layer was further extracted with EtOAc (6 x 25 mL) and then dichioromethane (3 x 25 mL). The organics were dried over MgS0₄, filtered, and concentrated in vacuo to afford 2-chloro-4-(4- methylpiperazin-l-y!)aniime (157 mg, 43%) as a light brown solid.

[00590] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 241. All compounds were characterized by LCMS.

Example 242. Synthesis of (Z)-fert-butyl 5-chloro-3-((2,5dioxoimidazolidin-4- ylidine)methyl>pyrazolori,5- lpyrimidine-7-vl(cvclopropvl)carbamate

[00591] 7¾rt-butyl 5-chloro-3-forniylpyrazolo[ 1 ,5- ]pyrimidin-7-yl(cyclopropyl)carbamate (5,05 g, 15 mrnol) was dissolved in anhydrous THF (100 mL). Diethyl 2,5-dioxoimida.zolidin-4- ylphosphonate (5.33 g, 22.5 mrnol) and NaOt-bu (1 .87 g, 19.5 mrnol) were then added. After stirring for 3 d at 23 °C, additional diethyl 2,5-dioxoimidazolidin-4-ylphosphonate (3.5 g) and aOt-bu (1.44 g) were added. After stirring an additional 24 h, the volatiles were removed in vacuo. The residue was stirred for 4 h in i-PrOH (50 mL) and water (250 mL) and then filtered to afford crude er -butyl 5-chloro-3-((2,5dioxoimidazolidin-4-ylidine)methyi)pyrazolo[l ,5- ]pyrmiidine-7-yl(cyclopropyl)carbaniate (5.64 g, 90%) as a mixture of Z:E isomers (5.3: 1). The crude solid was diluted with /-PrOH (110 mL) and heated to reflux. The solution was filtered and then allowed to cool to afford (Z)-tert-butyl 5-ch[oro-3-((2,5dioxoimidazolidin-4- ylidine)methyl)pyrazolo[] ,5- ]pyrimidine-7-yl(cyclopropyl)carbamate (3.58 g, 57%) as a bright orange solid in two crops. 1H NMR (CDC1₃, 400 MHz) δ: 10.37 (bs, 1 H), 8.17 (s, 1 H), 8.02 (bs, I I I ). 6.81 (s, 1H), 6.60 (s, 1H), 3.26 (dddd, 1H, J 6.8, 6.8, 3.2, 3.2 Hz), 1.43 (s, 9H), 0.87-0.94 (m, 2H), 0.62-0.68 (m, 2H). LCMS (ES): >90% pure, m/z 419 [M+l f .

[00592] Diethyl 2,5-dioxoimidazoiidin-4-ylphosphonate was prepared according to the procedure of Meanweil, et al. J. Org. Chem. 1991, 56, 6897.

Example 243. Synthesis of (Z)-5~((5~chloro^

yl)methylene)imidazolidine-2,4-dione

[00593]

5-chloro-3-((2,5dioxoimidazolidin-4-ylidine)methyl)pyrazolo[l ,5- a]pyrimidine-7-yl(cyclopropyl)carbamate (3,20 g, 7.66 mrnol) was suspended in

dichlorometha e (30 mL). Trifluoroacetic acid (30 mL) was added slowly and the solution became homogeneous. After 1 h, the volatiles were removed in vacuo. The residue was triturated in Et₂0 (100 mL) and the bright yellow solid was filtered off to afford (Z)-5-((5- cMoro-7-(cyclopropylamino)pyrazolo[l,5-a]pyri

(2.42 g, 99%). LCMS (ES)i >90% pure, m/z 319 | M 1 | ^' .

Example 244. Synthesis of (Z)-A^f-(3-(7-{cyclopropylamjno)-3-({2.5-dioxoimidazolidin-4- ylidene)methyl)pyrazolo [ 1 ,5 - jpyrimidin- 5 -y la.mino)phenyl)acetamide

[00594] ( T -5-((5-chloro-7-(cyciopropylammo)pyrazolo[l,5- ]pyrimidin-3- y])methyiene)imidazolidine~2,4-dione (75 mg, 0.23 mmol) was suspended in 1,4-dioxane (1.6 mL). N-(3-aniinophenyl)acetamide (52 mg, 0.35 mmoi), Cs₂C0₃ (105 mg, 0,32 mmol), (±)- BINAP (9 mg, 0.06 mmol) and palladium(II) acetate (7 mg, 0.04 mmol) were then added. The mixture was sealed and irradiated at 120 °C for 30 min in the microwave. H₂0 (8 mL) was added and the precipitate was filtered off and dried. The crude residue was purified via flash column chromatography (2.5-3.5% MeOH/dichloromethane) to yield (Ζ)-Λ'-(3-(7- (cyclopropylanuno)-3-((2,5-dioxoimidazolidin-4-y^

yiamino)phenyl)acetamide (12 mg, 12%) as a bright yellow solid. LCMS (ES): >90% pure, m/z 433 I M · i ] .

[00595] The compounds described in the following table were prepared using chemistries similar to those exemplified in Examples 242 to 244. All compounds were characterized by LCMS. Table 43B shows the biological activities of the compounds listed in Table 43A.

Table 43 A.





Table 43B.

AB:

Compound CK2: IC50 PIM2: %inh MDAMB453 AB: BxPC3

2.5 μΜ IC50 (μΜ) IC50 (μΜ)

W38 <0.01 12.679 0.503 3.401

X38 <0.01 -14.929 0.817 7.128

Y38 <().() 1 6.404 > 30

Z38 <0.01 5.1 1 0.363 2.273

A39 <0.01 -5.524 1.483 3.348

B39 <0.01 42.988 2.309 2.183

C39 <0.01 4.463 27.171

D39 <0.01 2.776 6.141

E39 <0.01 12.058 1.745 2.347

F39 <().() 1 29.369 24.684 > 30

G39 <0.01 38.351 9.58 13.05

H39 <0.01 32.314 1.187 5,5

139 <0.01 -19.596 3.914 13,069

J39 <0.01 19.119 2.462 4.126

K39 <0.01 -2.822 1.179 14,376

L39 <0.01 53.751 1.132 4.028

M39 <0.01 -12.471 1.824 20.033 AB:

Compound CK2: IC50 PI 2: %inh MDAMB453 AB: BxPC3

2.5 μΜ IC50 (μΜ) IC50 (μΜ)

N39 <0.01 1.334 2.178

039 <0.01 2.901 5.71

P39 <0.01 1.012 3.518

Q39 <0.01 50.224 0.568 2.448

R39 <().() 1 7.228 15.996

S39 <0.1 3.1 18 5.462

T39 <0.01 5.657 13.157

U39 <0.01 16.983 13.301

V39 <0.01 15.34 0.503 3.722

W39 <0.01 20.319 < 0.12 <0.12

X39 <0.1 23.241 3.558 0.409

Y39 <().() 1 45.658 1.685 5.123

739 <0.01 32.912 10.761 11.535

A40 <0.01 61 .33 1.214 4.211

B40 <0.01 , I , 3 ^ 8.026 > 30

C40 <0.01 54.631 0.568 0.427

D40 <0.01 56.161 3.885 4.838

E40 <0.01 45.415 10.016 7.671

F40 <().() 1 74.699 1.487 2.566

G40 <0.01 74.328 16.589 14.183

H40 <0.01 55.726 1.099 4.157

140 <0.01 67.726 0.882 2 795

J40 <0.01 69.269 14.73 5.949 40 <0.01 66.943 1.042 3.053

L40 <0.01 37.998 6.073 1.762

M40 <().() 1 26.534 2.111 1.273

N40 <0.1 23.553 3.706 1.379

O40 <0.01 60.445 16.1 19 > 30

P40 O.01 56.901 28.665 > 30

Q40 <0.01 41.399 2.892 5.65

R40 <0.01 30.569 3.021 4.466 AB:

Compound CK2: IC50 PI 2: %inh MDAMB453 AB: BxPC3

2,5 μΜ IC50 (μΜ) IC50 (μΜ)

S40 <0.01 33.343 3.273 13.798

T40 <0.01 64.565 0.656 1.485

U40 <0.01 39.619 2.736 3.191

V40 <0.01 63.083 1.997 3.383

VV40 <().() 1 12.679 0.503 3.401

X40 <0.01 -14.929 0.817 7.128

Y40 <0.01 807 0.248 9.761

Z40 <0.01 55.427 0.999 3.866

A41 <0.01 41.66 2.51 1 11 -2

B41 <0.01 35.147 0.764 2.282

C41 <o.o i 24.356 5.447 12.691

D41 <().() 1 -6.887 3.168 3.25

E41 <0.01 - 12.19 1.04 1 .794

F41 <0.01 19.291 15.678 26.981

G41 <0.01 73.187 0.803 9.037

H41 <0.01 76.943 0.852 5.263

141 <0.01 59.089 0.776 9.983

Example 245. Synthesis of (Z)-4-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4- yljdene)niethvj )pyrazolo[ l ,5- ]pyrimidin-5-yja.nimo)-N-i2-(di

-trifluoroacetate

[00596] (Z)-4-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazo[o[l,5-a]pyrimidin-5-ylamino)-3-fluoroberLzoic acid (25 mg, 0.06 mmoi) was suspended in DMF (0,2 mL). EDCI (13 mg, 0.7 mmol), HOBt (1 1 rng, 0.7 mmol), triethylamine (10 yL, 0.7 mmol), and iV, N-diethylethyienedimiine (8 ,uL, 0.7 mmol) were added sequentially. The reaction was heated to 65 °C. After 1 h, the solution was diluted with D SO (1 mL) and purified by reverse phase HPLC to yield (Z)-4-(7-(cyclopropylammo)-3-((2,5- dioxoimidazolidin~4-ylidene)niethyl)pyra^

(diethylamino)ethyl)-3-fluorobenzaniide 2,2,2-trifluoroacetate (29 mg, 76%). LCMS (ES): >90% pure, m/z 536 [M+l]⁺.

[00597 J The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 245. All compounds were characterized by LCMS. Table 44B shows the biological activities of the compounds listed in Table 44A.

Table 44A.

Table 44B.

Example 246. Syntheis of 4-(7-(cyclopropylamino)pyrazoio[l,5-a]pyrimidin-5- ylamino)benzoiiitrile hydrochloride

[00598] 5-chloro-N~cyclopropylpyrazolo[l ,5- ]pyrimidin-7-amine (208 mg, 1 mmol) was suspended in EtOH (1 mL). 4-aminobenzonitrile (236 nig, 2 mmol) and then cone. HCl (125 fiL, 1.5 mmol) were added and the reaction was placed in a 95 °C oil bath. After 24 h, additional cone. HCl was added (62 μΕ). After an additional 24 h, the reaction was cooled to 23 C'C and the filter cake was washed with EtOH (2 mL) to afford 4-(7-

(cyclopropylamino)pyrazolo[ 1 ,5-a]pyrimidin-5-yiamino)benzonitrile hydrochloride (205 mg, 63%) as a light brown solid. LCMS (ES): >90% pure, m/z 291 [M+l . Synthesis of 4-(7-(eyclopropylamino)-3-fomiylpyrazolo[ 1 ,5- ]pyrimidin-5- yla ino )bertzorti trile

[00599] 4-(7-(cyclopropylamino)pyrazolo[l .5-a]pyrimidin-5-ylamino)benzoni trile hydrochloride (205 mg, 0.62 mmol) was dissolved in anhydrous DMF (1 mL) and the solution was cooled to 0 °C by an external ice bath. POCl₃ (1 15 _uL, 1.25 mmol) was added dropwise keeping the internal temperature <5 °C. After addition, the ice bath was removed. After 5 h, the solution was poured into H?0 (20 mL) and the pH was adjusted to 1 1 by the addition of 6N

NaOH, The solution was allowed to stir for 1 h, and the precipitate was filtered off. The crude product was triturated with EtOH ( 7 mL), filtered, and dried under high vacuum ( 1 mmHg) to provide 4-(7-(cyclopropylamino)-3-fomiylpyr^

(129 mg, 64%) as an orange solid. LCMS (ES): >90% pure, m/z 319 j M · 1 j .

48. Synthesis of ( )-4-(7-(cyclopropylamino

ylidene)methyl)pyrazolo [1 ,5 - lpyrimidin-5 -ylamino)benzoni trile

[00600 J 4-(7-(cyciopropylamino)-3-formylpyrazolo[l ,5-a]pyrimidin-5-ylamino)benzonitriie (75 mg, 0.24 mmol) was suspended in EtOH (2.4 mL). Hydantoin (36 mg, 0.35 mmol) and piperidine (36 μί, 0.35 mmol) were added and the reaction was heated to 80 °C. After 15 h, the solution was filtered while warm and the filter cake was washed with warm EtOH (3 mL) to give (Z)~4~(7-(cyelopropylanimo)~3-((2,5-dw^ a]pyrimidin-5-ylammo)henzomtrile (76 mg, 80%) as a bright yellow solid. LCMS (ES): >90% pure, m/z 401 [M+l]⁺.

[00601] The compounds described in the following table were prepared using chemistries similar to those exemplified in Examples 246 to 248. All compounds were characterized by LCMS. Table 45B shows the biological activities of the compounds listed in Table 45 A.

Table 45 A.

Table 45 B.

U41 <0.01 57.088 1.019 1.56

V41 <0.01 16.198 0.352 3.08

Example 249. Synthesis of fcrt-butyl 5-(2-bromo-4-cyanophenylamirio )-3-formyrpyrazolo[ 1 ,5- vrimulin-7-s l(cvcK>prop ΠαηΙηιιηαιο

[00602] Zerf-butyl 5-chloro-3-formylpyrazolo[ 1 ,5-a.]pyrimidin-7-yl(cyclopropyl)carbamate (225 mg, 0.67 mmol) and 4-amino-3-bromobenzonitrile ( 197 nig, 1 mmol) were dissolved in anhydrous THF (4.5 mL). Sodium ie/t-butoxide (96 mg, 1 mmol) was added in one portion. After 1.5 h, the reaction was poured into H₂0 (25 mL) and extracted with EtOAc (3 x 30 mL). The organics were washed with brine (1 x 100 mL), dried over MgS0₄, filtered and concentrated in vacuo. The tan solid was purified via flash column chromatography (30% EtOAc/hexanes) to provide f erf-butyl 5-(2-bromo-4-cyanophenylamino)-3-formylpyrazolo[ 1 ,5-a]pyrimidin-7- y](cyciopropyl)carbamate (106 mg, 32%) as a pale yellow solid. LCMS (ES): >90% pure, m/z 497 [ + l ]⁺.

Example 250. S nthesi s of 3 -bromo-4-Γ / -(cyclopropylamino)-3 -formylpyrazolo [1 ,5- -5-ylamino)benzonitrile

[00603] 7¾rf-butyl 5 -(2-bromo-4-cyanophenylaniino)-3 -formylpyrazolo [1,5 -«]pyrimidiii-7- yl(cyclopropyl)carbamate (105 mg, 0.21 mmol) was dissolved in dich!oromethane (2 mL) and trifluoroacetic acid (2 mL) was added. After 1 h, the reaction was concentrated to dryness and the residue was triturated with Et₂0. The yello solid was collected and dried to give 3-bromo- 4-(7-(cyclopropylamino)-3-formylp 2,2,2- trifluoroacetate (72 mg, 67%).

Example 251. Synthesis of (Z)-3-bromo-4-(7-(cyciopropylaniino)-3-((2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazolo [1,5 - ]pyrimidin-5 -ylamino)benzonitrile

[00604] 4-(7-(cyclopropylammo)-3-formylpyrazolo^

2,2,2-trifiuroacetate (72 mg, 0.14 mmol) was suspended in EtOH (2.4 mL). Hydantoin (17 mg, 0,17 mmol) and piperdine (33 μΐ., 0.34 mmol) were added and the reaction was heated to 80 °C. After 15 h, the solution was filtered while warm and the filter cake was washed with warm EtOH (3 mL) to give (2)-3-bromo-4-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4- ylidene)methyl)pyrazolo[l,5-a]pyrimidin-5-ylamino)beiizonitrile (60 mg, 89%) as a bright yellow solid. LCMS (ES): >90% pure, m/z 479 | M · 1 ] .

[00605] The compounds described in the following table were prepared using chemistries similar to those exemplified in Examples 249 to 251. All compounds were characterized by LCMS. Table 46B shows the biological activities of the compounds listed in Table 46A.

Table 46 A.

Table 46B.

Example 252, Symhcsis of (^)-4-( 7-( c ciopi' px lai siiso )-3-{ ( i ~n^cU?yi-2.5-dioxom:ida/oiidii:-4- ylidene)methyl)pyrazolo [ 1 ,5 -a]pyrimidin- -y lamino )benzonitrile

[00606] 3-methylimidazolidine-2,4-dione was prepared according to the literature procedure setforth in Eur. JOC 2002, 1763.

[00607] 4-(7-(cyclopropy[arrnno)-3-formylpyrazolo[l,5-a]pyrimidin-5-ylaniino)ber^

(51 mg, 0.16 mmol) was suspended in EtOH (1.6 mL). 3-methylirmdazolidine~2,4~dione (28 mg, 0.24 mmol) and piperdine (24 μΐ,, 0.24 mmol) were added and the reaction was heated to 80 °C, After 15 h, the solution was diluted with H2O (2 mL) and filtered. The filter cake was washed with 50°/» H2O/50% EtOH (3 mL) and then dried in vacuo (-ImmHg) to furnish (Z)-4- (7-(cycloprop}4amino)-3-((l-methyl~2,5-dioxoimidazolidin-4~ylidene)

a]pyrimidm-5-ylarmno)benzonitril6 (45 mg, 68%) as a bright yellow solid. LCMS (ES): >90% pure, m/z 415 [M+l]⁺.

[00608] The compounds described in the following table were prepared using chemistries similar to those exemplified in Example 252. All compounds were characterized by LCMS. Table 4733 shows the biological activities of the compounds listed in Table 47 A.

Table 47B.

Exam le 253. Synthesis of (¾

ylidene)methyl)pyrazolo[l,5- 1pyriiTiidin--5-yloxy)beiizonitrile

[00609] (Z)-tert-bnty\ 5-ch.loro-3-((2,5-dioxoimidazoiidin-4-ylidene)methyl)pyrazolo[l,5- a]pyriniidiii-7-yl(cyclopropyl)carbamate (75 nig, 0, 18 mmol) was dissolved in anhydrous DMF (0.6 mL). 3-ch!oro-4-hydroxybenzonitrile (41 mg, 0.27 mmol) and K₂CO₃ (75 nig, 0.54 mmol) were added. After 24 h, H₂0 (3.5 mL) was added to the reaction and the bright yellow precipitate was filtered and dried. The crude solid was dissolved in dichloromethane (1 mL) and trifluroacetic acid (1 mL), After 1 h, the reaction was concentrated to dryness and the residue was triturated with Et₂0 (3 mL) and filtered to provide (Z)-3-chloro-4-(7-(cyclopropylamino)-3- ((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[ 1 ,5- ]pyrimidin-5-yloxy)benzonitrile (45 mg, 57% over two steps) as a bright yellow solid. Example 254, Synthesis of (Z)-5 -¾ { 5 -(5 -chloro-2-fluorophenylaniino)-7- fc cl propylam.ino)p azolo[^'l ,5-a]pyrirnidin-3-yl methyl.ene -3- -2,4-dione

[0061 Oj To (Z)-5-f(5-(5-chloro-2-t1uorophenylamino)-7-(cyelopropylamino)pyrazolo[l ,5- a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (500 mg, 1.17 mmol) in acetonitrile (15 mL) and pyridine (1.5 mL) was added formaldehyde (37% aq) (5.0 mL). The reaction mixture was stirred at 65°C for 5 minutes. Cooled to room temperature and filtered off the resulting solid. Washed with water and dried under vacuum to provide 450 mg (84%) of (Z)-5-((5-(5- ch[oro-2-fluorophenylamino)-7-(cycloprop

3-(hydroxyniethy])imidazolidine~2,4-dione as a yellow solid. LCMS (ES): >95% pure, m/z 458 [M+ l]⁺.

Example 255. Synthesis of (Z)-5 -((4-((5 -(5 -chl oro-2 -fluoropheny lam

(cyclopropylamino)pyrazolofl_^^

-5 -oxopentanoi c acid

[00611] To (Z)-5-((5-(5-chloro-2-fiuorophenylamino)-7-(cyclopropylaniino)pyrazolo[ 1 ,5- a]pyrimidin-3-yl)methylene)-3~(hydroxym (100 mg, 0.218 mmol) in pyridine (4.5 mL) was added glutaric anhydride (125 nig, 1.095 mmol), and DMAP (3 mg, 0.022 mmol). The reaction mixture was stirred at 75°C overnight after which the reaction was not complete. Added glutaric anhydride (125 mg, 1.095 mmol) and DMAP (3 mg, 0.022 mmol) and stirred an additional 16 hours at 75°C. Cooled to 0°C in ice bath and added 6M HQ until pH was less than 3 by pH paper. Filtered off the solid and washed with O.I M HQ. Dried under vacuum to provide 40 mg (32%) of (Z)-5-((4-((5-(5-chloro-2-fluorophenylamino)-7- (cyclopropylamino)pyrazolo[ 1 ,5~a]pyrimidin-3-yl)methykine)-2,5-dioxoimida.zoiidin- 1 - yl)methoxy)-5-oxopentanoic acid as a yellow solid, LCMS (ES): >95% pure, m/z 572 [M+l]^"\

Example 256. Synthesis of (Z -(4-((5-(5-chloro-2-fluorophenylamino)-7- (cyclopropylamino)pyrazolo[l,5-a]pyrim ^

-(4-methylpiperazin-l-yl)propanoate

[00612] To (Z)-5-((5-(5-cWoro-2-fluOTophenylamino)-7-(cyclopropyla.mino)pyrazolo[l,5- a]pyriniidin-3-yi)methylene)-3-(hydroxymethyl)imidazolidine-2,4-dione (100 mg, 0.218 mmol) in DMF (3 mL) was added 3-(4-methylpiperazin-l-yl)propanoic acid (75 mg, 0.436 mmol), dicyclohexylcarbodiimi.de (90 mg, 0.436 mmol), and DMAP (4.0 mg, 0.33 mmol). The reaction mixture was stirred at room temperature overnight and then diluted with ethyl acetate and washed I X water, 3X brine. The organic layer was dried with MgS0₄, filtered and adsorbed onto silica gel. The crude material was purified by column chromatography eluting with 0-10% MeOH/CH₂Cl₂ gradient. Pure fractions were combined and the solvent was removed. This material was crystallized from ethyl acetate and tiexane to provide 35 mg (26%) of (Z)-(4-((5- (5-chloro-2-fluorophenylamino)-7-(cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3- yi)methylene)~2,5~dioxoimidazoiidin~l ~y{)methyl 3~(4-methylpiperazm~l-yl)propanoate as a yellow solid. LCMS (ES): >95% pure, m/z 612 [M+l f.

Example 257. Synthesis of (Z)-(4-((5-(5-chloro-2-¾orophenylamirio)-7-

(cyclopropylamino)pyrazolo[l ,5-a]pyrimidin-3-yl)methylene)-2,5- -aminoacetate

[00613] To (Z)-5-((5-(5-chloro-2-fluorophenylamino)-7-(cyclopropylamino)pyrazolo[l ,5- a]pyrimidin-3-yl)methylene)-3-(hydroxymethyl)imidazolidine-2,4-dione (100 mg, 0.218 mmol) in DMF (3 mL) was added Boc-Gly-OH (153 mg, 0.873 mmol), dicyclohexylcarbodiimide (180 mg, 0.873 mmol), and DMAP (1 3 mg, 0.109 mmol). The reaction mixture was stirred at room temperature overnight, then diluted with ethyl acetate and washed IX with 1M HCl followed by 3X brine. The organic layer was dried with MgS0₄, filtered and adsorbed onto silica gel. The crude material was purified by column chromatography eluting with 5-15% EtOAc/CH₂Cl₂ gradient. Pure fractions were combined and the solvent was removed. To the residue was added 4M HCl/dioxane (4 mL) and stirred at room temperature for 2h. Removed excess HCl/dioxane. To the residue was added diethyl ether and the suspension was sonicated. The resulting solid was filtered off and washed with diethyl ether. Dried under vacuum to provide 23 mg (21%) of (Z)-(4-((5-(5-chloro-2-fluorophenylammo)-7-(cyclopropylamino)p>Tazolo[l ,5-a]pyrimidin-3- yl)methylene)-2₅5-dioxoimidazolidin-l -yl)methyl 2-aminoacetate hydrogen chloride as a yellow solid. (LCMS (ES): >95% pure, m/z 515 [M+l .

[00614J The following molecules were prepared using chemistries similar to synthesis in examples above. All compounds were characterized by LCMS. Table 48B shows the biological activities of the compounds listed in Table 48 A. Table 48A.

Table 48B.

Example 258. Synthesis of 5-((5-(4-(l H-pyrazol-l-yl phenytamino)-7- (cyclopropylamino pyrazolo[l ,5-a]pyrim

[00 15] To (Z)-5 -((5 -(4-( 1 H-pyrazol- 1 -y !)pheny lamino)-7-(cyclopropy lamino)pyrazolo [1 ,5- a]pyrimidin-3-yl)methylene)imidazolidine-2,4-dione (40 mg, 0.091 mmol) in a Parr pressure reaction vessel was added acetic acid (6,0 mL) and 10% Pd/C (20 mg). The reaction vessel was placed on the Parr shaker at 55 psi for 3 days. Filtered through celite and purified by mass- directed LC/MS to provide 5-((5-(4-(l H-pyrazol-l-y[)phenylamino)-7-

(cycloprcypylamino)pyrazo]o[l ,5-a]pyrimidin~3-yl)niethyl) as the TFA salt. LCMS (ES): >95% pure, m/z 444 [M+lf . Example 259, Synthesis of 2-^^^

ylaminolphenol

[00616] To 5-chloro-N-cyclopropylpyrazolo[l ,5-a]pyrimidin-7-amine (500 mg, 2.396 mmol) in EtOH (10 mL) was added 4-amino-2-chlorophenol (516 mg, 3.59 mmol) followed by concentrated HCl (0.218 mL, 2.64 mmol). The reaction mixture was stirred at reflux temperature for 4 days. Removed 5 mL of EtOH on rotavap followed by addition of 5 mL of diethyl ether. The resulting solid was filtered off and rinsed with diethyl ether, Dried under nitrogen to provide 582 mg (77%) of 2-chloro-4-(7-(cyclopropylamino)pyrazolo[l ,5- a]pyrimidm-5-ylamino)phenol as the HCl salt. LCMS (ES): >95% pure, m/z 316 [M+1J⁺.

Example 260, Synthesis of S-O-cMoro^-hydroxYphenviamino^^- fcyclopropylammo^yrazo o^.S-ajpyrimidine-B-carfa^

[00617] To 2-chloro-4-(7-(cycl propylamino)pyrazolo[l,5-a]pyrirriidin-5-ylamino)phenol (582 mg, 1.84 mmol) in DMF (4.5 mL) cooled to 0°C was added phosphorous oxychloride (0, 14 mL, 5.52 mmol) dropwise. The reaction mixture was stirred while warming to 45°C over 3h. This was cooled down to 0°C and added slowly to an ice-cold solution of 2M aOH while stirring. Upon completion, the mixture was stirred at room temperature for Ih. The resulting solid was f ltered off and washed with water to provide 412 mg (65%) of 5-(3-chloro-4- hydroxypheiiylamino)-7-(cyciopropylamino)pyrazoio[l,5-a]pyrimidine-3-carbaldehy LCMS (ES): >95% pure, m/z 344 j M S f .

(cyclopropylammo)pyrazolo |^" l ,5-a pyrimidine-3-carbaldehyde

[00618] To 5-(3~chioro~4 iydroxyphenylamir ; ~7^ ,5- a]pyrimidine-3-carbaldehyde (100 mg, 0,291 mmol) in DMF (3 mL) was added K₂CO₃ (100 mg, 0.727 mmol). The reaction mixture was stirred at 80°C for 16h. Diluted with EtOAc and extracted into 2M HC1. The aqueous layer was basified to pH of 14 with 2M NaOH and extracted 2X with EtOAc. The organic layer was washed 3X with brine and dried with MgS0₄, Filtered and purified by column chromatography eluting with 10%-30% MeOH/EtOAc gradient. Combined pure fractions to provide 45 mg (36%) of 5-(3-chloro-4-(3- (dimethylamino)propoxy)phenylamm^

carbaidehyde LCMS (ES): m/z 429 [M+l]⁺. Example 262. Synthesis of (Z)-5-((5-(3-chloro-4-(3-(dimethylamino)propoxy)phenylamino)-7- (cyclopropylamino) pyrazoloi 1 ,5-alpvrimidin-3-vl)methylene)imidazolidine-2,4-dione

[00619 J To 5-(3-chloro-4-(3-(dimethylamino)propoxy)phenylamino)-7- (cyclopropylamino)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (79 mg, 0.184 mmol) in EtOH (3 mL) was added piperidine (22uL 0.221 mmol) followed by hydantoin (21 mg, 0.202 mmol). The reaction mixture was stirred at 85°C for 6h. The solvent was removed by rotary evaporation and the residue was diluted with 3 mL of water. The suspension was sonicated and the resulting solid was filtered off and washed with water followed by a 1 : 1 mixture of EtOH/water. The material was Dried under vacuum to provide 62 mg of (Z)-5-((5-(3-chloro-4-(3- (dimethylamino)propoxy)phenylam

yl)methylene)imidazolidine-2,4-dione LCMS (ES): m/z 511 [M+l]^""^".

[00620] The compounds listed in Table 49A were prepared according to the procedures described abo ve. Table 49B sho ws the biological activities of the compounds listed in Table 49A.

Table 49A.

Table 49B.

Example 263, Synthesis of 7-(cvcIopropvlamino -5-(methvlthio)pvrazololl,5-ali>vrimidiiie-

3-carbaldehyde

[00621] To 5-chloro-7-(cyclopropylamino) pyrazolo [1, 5 -a] pyrimidine-3-carbaldehyde (4.0g, 16,87 mmol) in dimethylformamide was added sodium thiomethoxide (3.54 g, 50.5 mmol) and the reaction mixture was heated to 80°C for 2 hrs. Cooled the reaction mixture, added water, stirred for 15 minutes and filtered white precipitate, dried to yield 7- (cyclopropylamino)-5-(methylthio)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (3.60 g, 86% yield), LCMS (M+l=249)

Example 264. Synthesis of fZ)-5-((7-(cyclopropylanimo)-5-(methy

[00622 | The above product 7-(cyc!opropyiamino)-5-(methySthio) pyrazolo [1 , 5-a] pyrimidine-3-carbaldehyde was dissolved in 20,0 mL ethanol, added hydantoin (2.82 g, 28, 17 mmol) and piperidine (2.70 mL). The reaction was heated to 80°C for overnight. Cooled the reaction mixture and yellow precipitate was filtered, washed with ethanol, dried to yield (Z)-5- ((7-(cyclopropylamino)-5-(niethylthio) pyrazolo [1 , 5-a] pyrimidin-3-y!) methylene)

imidazolidine-2, 4~dione 4.18 g (90% yield). LCMS (M+l=331)

I 0 Example 265. Synthesis of (Z)-5-((7-( cyclopropylamino)-5-methylsulfonyl)pyrazolo[ 1 ,5 alpyrimidm-3-yl methylene imidazolidine-2,4-dione

[00623] To (Z)-5-((7-(cyclopropylamino)-5-(methylthio) pyrazolo [1 , 5-a] pyrimidin-3-yi) methylene) imidazolidine-2, 4-dione (step b) (4.2 g, 12.68 mniol) in 40.0 mL dichloromethane, was added meta-Chloroperoxybeiizoic acid (8.75 g. 50.7 mmols) and the reaction mixture was stirred at room temperature overnight. Added another 1 0.0 mL of dichloromethane, sonicated for 10 minutes and then filtered the yellow precipitate to yield (Z)-5-((7-(cyclopropylamino)-5- (methyisulfonyl) pyrazolo [1 , 5~a] pyrimidin-3-yl) methylene) imidazolidine-2, 4-dione (3.7 g, 73% yield). LCMS (M+ 1=363)

Example 266. Synthesis of { S,Z)-5-((5-(l -{ ^'3-chlorophenyl)ethylammo)-7- {cvclopropyiamino)pyrazo1o[ ^'l ,5-a]pyrimidin-3-yl methyl.ene imidazolidme-2,4-dione

[00624] To (Z)-5-((7-(cyclopropyiarnino)-5-(methyisulfonyl) pyrazolo [1 , 5-a] pyrimidin-3- yl ) methylene) imidazolidine-2, 4-dione (step c) (10 mg, 0.0275 mmol) in 200 uL NMP, was added (S)-l ~(3~chlorophenyi) ethanamine (23.2 ul, 0.165 mmols) and the reaction mixture was heated in the microwave at 120 °C for 20 minutes. The mixture was concentrated and diluted with MeOH and purified by preparative HPLC to yield (S,Z)-5~((5-(l-(3- chiorophenyl)ethyiamino)-7-(cyclopropylamino)pyrazolo[l,5-a]pyrimidin-3- yl)metliylene)imidazolidine-2,4-dione LCMS (M+ 1 =438)

[00625] The benzyl amine analogs shown below were prepared using procedures exemplified above or methods previously described in Examples 27, 28, and 29. Table SOB shows the biological activities of the compounds listed in Table 50A.

Table 50A.

Table 5 OB,

AB:

Compound CK2: IC50 PI 2: %inh MDAMB453 AB: BxPC3

(iiM) 2.5 uM IC50 ( s M) IC50 (u M)

Z42 <0.1 46.188 3.333 6.124

A43 <0.1 51.937 5.542 11.672

B43 <0.1 57.05

C43 <0.01 46.18 3.788 10.91

D43 <0.1 46.258 18.532 21 .512

E43 <0.1 35.462 3.442 7.676

F43 <{),] 47.13 5.127 11.302

G43 <0.1 40.238 0.25 0.311

H43 <0.01 70.122 1.575 7.156

143 <0.1 44.879

J43 <0.1 54.24

K43 <0.1 59.084

L43 <0.01 54.602 7.624 11.881

M43 <0.01 58.527 6.073 4.019

N43 <0.01 53.361 3.747 5.688

043 <0.1 39.481 7.183 13.677

P43 <0.01 30.394 7.84 10.566

Q43 <0.01 20.98 7.9 11.155

R43 <0.1 13.354 6.761 13.999

S43 <0.1 37.578 14.925 25.375

T43 <{),] 8.871

U43 <0.01 37.958 14 ,44 / 23,012

V43 <0.01 31.461 5.943 8.756

W43 <0.01 45.881 4.407 21.36

X43 <0.1 18.114 4.988 24.742

Y43 <0.1 37.959 4.445 15.629

Z43 <0.01 36.842 4.787 22.226

A44 <0.01 22.798 20.301

B44 <0.1 24.036 22.599 > 30

C44 <0.01 43.464 4.882 6.246

D44 <0.1 48.152 16.668 14 AB:

Compound CK2: IC50 PI 2: %inh MDAMB453 AB: BxPC3

(iiM) 2.5 uM IC50 ( s M) IC50 (u M)

E44 <0.1 46.863 10.367 10.084

F44 <0.1 20.409

G44 <0.01 24.063 1.604 7.202

H44 <0.1 28.124 4.276 8.203

144 <0.1 31.932 6.52 14.085

J44 <0.1 35.323 6.219 17.199 44 <0.01 84.937 3.914 3.632

L44 <0.1 40.527

M44 <0.01 31.316 5.907 10.674

N44 <0.1 24.588 4.282 5.226

044 <0.1 18.817

P44 <0.1 14.088

Q44 <0.1 26.537

R44 <0.01 41.193 22.212 20.611

S44 <0.1 12.628 12.315 20.837

T44 <0.1 9.802

U44 <0.1 34.87

V44 <0.1 14.908 5.184 10.102

W44 <().() 1 40.252 0.172 0.242

X44 <0.01 20.632 6.096 10.292

Y44 <0.01 36.847 2.301 1.384

Z44 <0.01 42.536 1.528 4.51

A45 <0.01 28.395 1.067 4.558

B45 <0.01 12.045 1.004 1.612

C45 <0.1 -75.739 5.504 9.198

D45 <0.1 -9.176 <0.12 <0.12

E45 <0.01 1 1 .678 3.938 11.216

F45 <0.1 -19.013 5.844 7.288

G45 <0.01 2.857 4.311

H45 <0.01 45.857 16.617 13.54

145 <0.01 2.447 7.898 AB:

Compound CK2: IC50 PI 2: %inh MDAMB453 AB: BxPC3

(iiM) 2.5 uM IC50 ( s M) IC50 (u M)

J45 <0.1 77.85 0.573 3.391

K45 <0.01 30.57 5.471 9.4

L45 <0.01 54.081 4.999 3.386

M45 <0.01 75.785 5.792 28.828

N45 <().() 1 62.228 5.556 13.076

045 <0.1 46.251

P45 <0.01 74.254 2.905 10.582

Q45 <0.1 74.048 14.792 > 30

R45 <0.01 28.85 3.477 5.733

S45 <0.01 32.937 2.98 7.51 1

T45 <0.1 -6.871 21.079 14.555

U45 <0.1 38.332 1.063 1.443

V45 <0.1 17.03 3.83 19.496

W45 <0.1 57.926 2.214 15.246

X45 <0.1 33.091

Y45 <0.1 20.409

Z45 <0.1 48.01

A46 <0.1 41.388 1 1.35 3.499

Example 267. Synthesis of^*5,7-dich.loro-6-methylpyrazoio!^"l,5-alpyrimidine

[00626] Under nitrogen gas atmosphere, sodium (3.5 g, 151 mmol) was added to ethanoi (125 mL) in small portions and stirred at room temperature until all the sodium had dissolved. A solution of 3-aminopyrazole (12.5 g, 150 mmol) in ethanoi (20 mL) and diethyl methylmalonate (26 mL, 1 3 mmol) were dropped, successively, to the above solution. The mixture was refluxed at 90°C for 10 hours, cooled to room temperature, and filtered under vacuum. To the solid, cold 5N HQ was added and the resulting solid was collected by filtration under vacuum. The intermediate, 6-methylpyrazolo[l ,5-a]pyrimidine-5,7-diol, was recovered as an off-white solid in 72% yield (17.9g). This material was used for the next step without further purification, LCMS (M+l=166)

[00627] Under nitrogen gas atmosphere, phosphorous oxychloride (160 mL, 1 ,72 mol) and dimethyl aniline (16 mL, 132 mmol) was added successively to the intermediate prepared above (16 g, 97 mmol), The mixture was heated at 110°C for 4 hours then excess POCL was removed under vacuum. The residue was made basic with 3N NaOH solution (pH = 9-10) and extracted with ethyl acetate (3x). The combined organic layers were dried over anhydrous Na₂S0₄, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (100% DCM) to provide 15.8 grams of the solid yellow product, 5,7-dichloro-6- methylpyrazolo[l ,5-a]pyTimidine (81 % yield). LCMS i fvl ! 203 }

Example 268, Synthesis of 5-chloro-7-(c clopropylamino -6-methylpyrazolori.5-a]pyrimidine-

[00628] To the reaction flask, 5,7-dichloro~6-methylpyrazolo[l ,5-a]pyrimidine (5 g, 25 mmoi) was added along with cyclopropyl amine (1.8 mL, 25 mmol), triethylamine (3.5 mL, 25 mmol), and acetonitrile (87 mL). The reaction was stirred at room temperature for 3 hours then heated at 85°C for an additional 6 hours. The mixture was cooled to room temperature, diluted with water, filtered and washed with water. The intermediate, 5-chloro-N-cyclopropyl-6- methylpyrazoio[l ,5-a]pyrimidin-7-amine, was further purified by silica gel chromatography (10% ethyl acetate/^'hexanes) to provide 4.8 grams of a white solid (86% yield). LCMS (M+l= 223)

[00629] To the intermediate (3.6 g, 16 mmol) isolated above in DMF (59 mL) was added phosphorous oxychloride (9 mL, 96 mmol) slowly at room temperature. The reaction mixture was allowed to stir at room temperature for 10 hours then quenched by addition to 6N NaOH solution. The pH of the mixture was adjusted with 6N HC1 to pH = 7-9. The solid was recovered by filtration and washed with water. The product, 5-chloro-7-(cyclopropylamino)-6- methylpyrazolo[l,5-a]pyrimidine-3-carbaldehyde, was purified by recrystallization from ethyl acetate/hexanes to yield a white solid in 73% yield (2,9 g). LCMS (M 1 251) 269. Synthesis of tert-butyl 5-chloro-3-formyl-6-methylpyrazolo[ 1 ,5-a]pyriniidin-7- ylfcyclopropypcarbama

To 5 -chloro-7-(cyclopropylamino)-6-methylpyrazo io [ 1 , 5 -a]pyrimidine-3 - carbaldehyde (2.9 g, 11.7 mmol) in methylene chloride (22 mL) was added triethylamine (2 ml 14 mmol), dimethyl aminopyridine (100 mg, 0.8 mmol), and di-t-butyldicarbonate (3,1 g, 14 mmol). The mixture was stirred at room temperature for 10 hours. The reaction mixture was transferred to a separately funnel, washed I X with H₂0, 2X with brine, dried over MgS0 , filtered, and evaporated to dryness to provide an oily residue. The crude material was purified by silica gel chromatography (25% ethyl acetate/hexanes) to yield a light orange solid (3.6 g, 88% yield), tert-butyl 5-chloro-3-formyl~6-methylpyrazolo[ 1 ,5-a]pyrimidin-7- yl(cyclopropyl)carbamate. LCMS (M+l= 351)

-5-yiamino benzonitrile

[00631] To 4-amino-3-chlorobenzonitrile (52 mg, 0.34 mmol), Cs₂C0₃ (130 mg, 0.4 mmol) were added to tert-butyl 5-chloro-3-formyl-6-methylpyrazolo[l,5-a]pyrimidin-7- yl(cyclopropyl)carbamate (100 mg, 0.29 mmol) dissolved in 1,4-dioxane (1.1 mL). Racemic BINAP (11 mg, 0.017 mmol) and paliadium(II) acetate (8 mg, 0.011 mmol) were then added. The mixture was sealed and irradiated at 1 10 °C for 60 min in the microwave. Et₂0 (3 mL) was added and the solution was filtered. The filtrate was concentrated in vacuo. The crude residue was dissolved in dichloroniethane (1.5 mL) and trifluoroacetic acid ( 1.5 mL). After stirring at room temperature for 1 hour, the solution was concentrated under a stream of air, The crude material was purified by silica gel chromatography (3% acetone/dichloromethane) to yield the product, 3-chloro-4-(7-(cyclopropylamino)-3-f miyl-6-methylpyrazolo[l,5-a]pyrimidin-5- ylamino)benzonitrile (34 mg, 33% yield). LCMS (M+l.=367)

Example 271 , Synthesis of 3-chloro-4-(7-(cyclopropyiamino)-3-((2,5-dioxoimidazolidm-4- ylidene)methyl)-6-meihylpyr3zolo[l,5-alpyrimidin-5-ylamino)benzonitrile

[00632] Hydantoin (2,7 mg, 0.027 rnmol) and 3-chloro-4-(7-(cyclopropylarn.ino)-3-formyl-6- methylpyrazolo[l,5-a]pyrimidin-5-ylamino)benzonitrile (10 mg, 0.027 rnmol) were dissolved in ethanol (0.4 mL) along with piperidine (3 uL, 0.03 rnmol). The reaction was heated at 80 °C. After 10 hours, the reaction was cooled to r.t, diluted with water, and the precipitate was collected and washed with water, 1 : 1 ethanobwater, then ethanol. The bright yellow solid was dried in vacuo to give 3-chloro-4-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4- ylidene)methyl)-6ntnethylpyrazolo[ 1 ,5-a]pyrimidin-5-ylamino)benzonitrile (7 mg, 58% yield). LCMS ( · i 449}

[00633] The following molecules were prepared using chemistries similar to synthesis in examples above. All compounds were characterized by LCMS. Table 5 I B shows the biological activities of the compounds listed in Table 51 A.

Table 51 A.

Table 5 IB.

Example 272, S ynthesis of -chloro-7-( c yclopropyl ammo)- 2-methylpyrazol o Γ 1 , -a] yrimidine- -carbaldehvde

[00634] To the reaction flask, 5,7-dichloro-2-methylpyrazolo[l,5-a]pyrimidine (2 g, 10 mmol) was added along with cyclopropyS amine (0.7 mL, 10 mmol), triethy!amine (1.4 mL, 10 mmol), and acetonitrile (30 mL). The reaction was stirred at room temperature for 8 hours then cooled to room temperature, diluted with water, filtered and washed with water. The

intermediate, 5-chioro-N-cyclopropyi-2~methylpyrazolo[l ,5-a]pyrimidiii-7-amiiie, was dried under vacuum to provide 1.85 grams of a white solid (83% yield). LCMS (M+l ^::: 223)

[00635] To the intermediate (1.9 g, 8.3 mmol) isolated above in DMF (31 mL) was added phosphorous oxychloride (4.6 mL, 49.7 mmol) slowly at room temperature. The reaction mixture was allowed to stir at room temperature for 10 hours then quenched by addition to 6N NaOH solution. The pH of the mixture was adjusted with 6N HC1 to pH ^::: 7-9. The solid was recovered by filtration and washed with water. The product, 5-chloro-7-(cyclopropylamino)-2- rnethylpyrazolo[l ,5-a]pyrimidme~3~carbaldehyde, was obtained as a white solid in 80% yield (1 .7 g). LCMS ( M i 25 \ )

Example 273. Synthesi s of tert-buty 1 5 -chloro-3 -formy 1-2-methylpyrazolo [1 ,5 -ajpyrimi din-7- yl(cyclopropyl)carbamate

[00636] To 5-chloro-7-(cyclopropylamino)-2-methylpyrazolo[l ,5-a]pyrimidine-3- carbaldehyde (1.7 g, 6.7 mmol) in methylene chloride (13 mL) was added triethylamiiie (1.1 mL, 8 mmol), dimethylaminopyridine ( 100 nig, 0.8 mmol ), and di-t-butyldicarbonate (1 .8 g, 8 mmol). The mixture was stirred at room temperature for 10 hours. The reaction mixture was transferred to a separator}' funnel, washed I X with H₂0, 2X with brine, dried over MgS0₄, filtered, and evaporated to dryness to provide an oily residue which solidified on standing. The product, tert-butyl 5-chloro-3-formyl-2-methylpyrazolo[ 1 ,5-a]pyrimidin-7- yl(cyclopropyl)carbamate, was recovered as an off- white solid in 82% yield (1.9g). LCMS (M+l = 351)

Example 274. Synthesis of 5-(4-( lH-pyrazol-l-yl)phenylamino)-7-(cyclopropylamino)-2- -a] pyrimidine-3 -carbai dehyde

[00637] To 4-(lH-pyrazo3-l-yl)aniline (54 mg, 0,34 mmoi), Cs₂C0₃ (130 mg, 0,4 mmol) were added to tert-butyl 5-chloro-3-formy[-2-methylpyrazolo[l ,5-a]pyrimidin-7- yl(cyclopropyl)carbamate (100 mg, 0.29 mmol) dissolved in 1 ,4-dioxane (1.1 mL). Racemic BINAP ( 1 1 mg, 0.017 mmol) and palladiumfll) acetate (8 mg, 0.01 1 mmol) were then added. The mixture was sealed and irradiated at 1 10 °C for 60 min in the microwave, Et₂Q (3 mL) was added and the solution was filtered. The filtrate was concentrated in vacuo. The crude residue was dissolved in dichloroniethane (1.5 mL) and trifluoroacetic acid (1.5 mL). After stirring at room temperature for 1 hour, the solution was concentrated under a stream of air. The crude material was purified by silica gel chromatography (15% acetone/dich!oromethane) to yield the product, 5-(4-(lH-pyrazol-l-yl)phenylamino)-7-(cyclopropylamino)-2-methyl^

a]pyrimidine-3-carbaldehyde (24 mg, 23% yield). LCMS (M · 1 374 )

Example 275. Synthesis of -; (5-{4-( i i l -pv ra/.ol- 1 -vi jphcnv ianiir > -7-{cvc k'pnipvian ii ' )-2- methylpyrazoloj^" ! ,5-a pyrimidin-3-yl)methylene)iniidazolidine-2,4-dione

[00638] Hydantoin (3 mg, 0.03 mmol) and 5-(4-(lH-pyrazol-l-yl)phenylamino)-7- (cyclopropy3.amino)-2-m.ethylpyrazolo[l ,5-a]pyrimidine-3~carbaldehyde (12 mg, 0.03 mmol) were dissolved in ethanol (0.4 mL) along with piperidine (3 uL, 0.03 mmol). The reaction was heated at 80 °C in the microwave for 2 hours. The reaction was then cooled to r,i, diluted with water, and the precipitate was collected and washed with water, 1 : 1 ethanobwater, then ethanol. The yellow solid was dried in vacuo to give 5-((5-(4-(lH-pyrazol- 1 -yl)phenylamiiio)-7- (cy clopropylarrimo)-2-rn ethylpyrazolo [ 1 ,5 -ajpyrimi din-3 -y I )methylene)imidazolidine-2,4-dione (9.5 mg, 65% yield). LCMS (M+ 1=456)

Table 52. Biological Activies of Example 275

76. Synthesis of 7-chloro-5-(methylthio)pyrazolo[ 1 ,5-a]pyrimidine-6-carbomtrile

[00639] Under nitrogen gas atmosphere, 2~cyano~3,3~bismethylthio-2-propenoic methyl ester (6 g, 29,5 mmol) was added to ethanol (40 mL) along with 3-aminopyrazole (2,6 g, 31 mmol) and the mixture was refiuxed for 2.5 hours. The reaction was then cooled to room temperature and precipitate was collected by filtration under vacuum. The solid was washed with ethanol and dried under vacuum to give 7-hydiOxy-5-(methyithio)pyrazolo[l,5-a]pyrimidine-6-- carbonitrile in 67% yield (4.1 g). This material was used for the next step without further purification, LCMS (M+ 1=207)

[00640] Under nitrogen gas atmosphere, phosphorous oxychloride (9.4 mL, 101.3 mmol) and dimethylaniline (2.6 mL, 20.3 mmol) was added successively to the intermediate, 7-hydroxy-5- (methyithio)pyrazolo[l ,5-a]pyrirmdine-6~carbonitriie, prepared above (4.1 g, 19.7 mmol). The mixture was heated at 1 10°C for 4 hours then excess POCI₃ was removed under vacuum. The residue was made basic with 3N NaOH solution (pH = 9-10) and extracted with ethyl acetate (3x). The combined organic layers were dried over anhydrous Na₂Si)4 , filtered, and

concentrated under vacuum. The residue was purified by recrystallization from ethyl acetate hexanes to provide the product, 7-chloro-5-(rnethylthio)pyrazoio[l,5-a]pyrirnidme-6-carbonitrile (80% yield). LCMS (M+ 1=225)

Example 277. Synthesis of 7-(cyclopropylammo)-3-formyl-5-(methylthio)pyrazolo[l ,5- ~6~carbonitrile

[00641] To the reaction flask, 7-cWoro-5-(methylthio)pyrazolo[l,5-a.]pyrimidine-6- carbonitrile (3.1 g, 13.7 mmol) was added along with cyclopropyl amine (0.96 mL, 13.7 mmol), triethylamine (1.9 mL, 13.7 mmol), and acetonitrile (30 mL). The reaction was stirred at 85°C for 10 hours then the mixture was cooled to room temperature, diluted with water, filtered and washed with water. The intermediate, 7-(cyclopropylamino)-5-(methyIthio)pyrazolo[ 1 ,5- a]pyrimidme-6-carbonitrile, was further purified by recrystallization from ethyl acetate hexanes to provide 3 grams in 89% yield. LCMS {M^■ 1 246)

[00642] To the intermediate (3 g, 12.2 mmol) isolated above in DMF (45 mL) was added phosphorous oxychloride ( 13.7 mL, 146 mmol) slowly at room temperature. The reaction mixture was allowed to stir at 70°C for 10 hours, cooled to room temperature, and quenched by addition to 6N NaOH solution. The pH of the mixture was adjusted with 6N HC1 to pH = 7-9. The solid was recovered by filtration and washed with water. The product, 7- (cyclopropylamino )-3 -formyl-5 -(methylthi o)pyrazo I o [ 1 ,5 -a] pyrimidine-6-carbonitri le, was obtained as a solid in 38% yield (1.28 g). LCMS { · i 274)

Example 278. Synthesis of 7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4-ylide¾ie

5-(methylsulfinyl)pyrazolo[

((2,5-dioxoiniidazolidin-4-ylidene)methyl)-5-(methylsulfonyl)p

carbonitrile

[00643] Hydantoin (366 mg, 3.7 mmol) and 7-(cyclopropylamino)-3-formyl-5- (methylthio)pyra.zolo[l ,5-a]pyrimidine-6-carbonitrile (1 g, 3.7 mmol) were dissolved in ethanoi (18.5 mL) along with piperidine (3.7 mL, 3,7 mmol). The reaction was heated at 80 °C. After 10 hours, the reaction was cooled to r.t, diluted with water, and the precipitate was collected and washed with water, 1 : 1 ethanoi: water, then ethanoi. The yellow solid was dried in vacuo to give the intermediate, 7-(cyclopropylamino)-3-((2,5-dioxoimidazolidm-4-ylidene)methyl)-5- (methylthio)pyrazolo[ 1 ,5-a]pyrimidine-6-carbonitrile (1.1 g, 83% yield), LCMS ( + 1=356)

[00644 J The intermediate (1.1 g, 3.04 mmol) was mixed with m-chloroperbenzoic acid (1 .9 g, 7.6 mmol) in dichloroniethane (12 mL). The mixture was allowed to stir at room temperature for 12 hours. The solid was collected by filtration, washed dichloroniethane then dried under vacuum overnight. The products, 7-(cyclopropylamino)-3-((2,5-dioxoimidazoiidiii-4~ ylidene)methyl)-5-(methylsulfonyl)pyrazolo[ 1 ,5-a]pyrimidine-6-carhoni rile and 7- (cyclopropylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)-5-

(methylsulfinyl)pyrazolo[l ,5-a]pyrimidine-6-carbonitrile, were recovered as a yellow solid in quantitative yield. LCMS (M+l = 372) and LCMS (M+l = 388) Example 279. Synthesis of 5-( 1 -f 3-chloi phenyl)ethylamin^

dioxoirn.idazoiidin-4-ylidene methyl^')pyrazolo[ L5-a]pyrimidine-6-carbonitrile

[00645] The mixture of 7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methy 5-(methylsu3ibnyl)pyra lo[l ,5-ajpyriraidme-6-carbonitri.le and 7-(cyclopropylamino)-3-((2,5- dioxoimidazolidin-4-ylidene)methyl)-5-(methyisulfinyl)pyrazoio[ 1 ,5-a]pyrimidine-6- carboiiitrile (20 mg) was mixed with (R)-l-(3-c-hlorophenyl)ethanamine (33 mg) in z-propanol (0.5 rtiL). The reaction mixture was heated at 90°C in the microwave for 1 hour. The reaction was cooled to room temperature and concentrated under vacuum. The residue was diluted with water, filtered and washed with water followed by 20% ethano!/water mixture. The solid was dried under high vacuum to give 2 mg of the product, 5-(l-(3-chlorophenyl)ethylamino)-7- (cyclopropylamino)-3-((2,5-dioxoim

carbonitrile. LCMS (M+l=463)

[00646] The following molecules were prepared using chemistries similar to synthesis in examples above. All compounds were characterized by LCMS, Table 53B shows the biological acti vities of the compo unds listed in Table 53 A. Table 53A.

Table 53B.

Example 280. Synthesis of 2-(4-(7-(cyclopropylamino)-3-formylpyrazolo[ 1 ,5~a]pyrimidm-5- yl)piperazin- 1 -yl)nicotinomtrile

[00647] In a reaction flask, 2-(piperazin-l-yl)nicotinonitriie (22 mg, 0.11 mmol) was mixed with 5-chloro-7-(cyc[opropylarnino)pyrazolo[l,5-a]pyrimidine-3-carbaldehyde (27 mg, 0.11 mmol) in DMF (0.5 ml.) along with potassium carbonate (32 mg, 0.23 mmol). The reaction was heated at 95°C for 12 hours then partitioned between water and ethyl acetate. The organic layer was washed with water then saturated NaCl solution. The ethyl acetate layer was isolated, dried of anhydrous sodium sulfate, fi ltered, and evaporated to dryness. The product, 2-(4-(7- (cyclopropylamino)-3-formylpyrazolo[ 1 ,5-a]pyrimidin-5-yi)piperazin- 1 -yljiiicotinonitrile, was recovered in 35% yield (16 mg) after recrystailization from ethyl acetate/hexanes. LCMS (M+l=389)

Example 281. Synthesis of 2-(4-(7-(cyciopropylamino)-3-((2,5-dioxoimidazolidin-4- viidene)methyl)pyrazolo j^" 1 ,5-a1pyrimidin-5-yl)piperazin- 1 -yDnicotinonitrile

[00648] Hydantoiii (4 mg, 0.04 mmol) and 2-(4-(7-(cyclopropylamino)-3- formylpyrazolo[l ,5-a]pyrimidin-5-yl)piperazin-l-yl)nicotinomtri ( 16 mg, 0,04 mmol) were dissolved in ethanol (0.5 mL) along with piperidine (4 uL, 0.04 mmol). The reaction was heated at 80 °C for 12 hours. The reaction was then cooled to r.t, diluted with water, and the precipitate was collected and washed with water, 1 :1 ethanohwater, then ethanol. The yellow solid was further purified by recrystailization from ethyl acetate/hexanes and dried in vacuo to give 2-(4-(7-(cyc[opropylamino)-3-((2,5-dioxoimidazolidin-4-ylidene)methyl)pyrazolo[l ,5- a]pyrimidin-5-yl)piperazin-l-yl)mcotinonitrile (2 mg, 21% yield). LCMS (M-t-l^:::471)

[00649] The following molecules were prepared using chemistries similar to synthesis in examples above. All compounds were characterized by LCMS. Table 54B shows the biological activities of the compounds listed in Table 54A. Table 54Λ.

Table 54B.

AB:

Compound CK2: IC50 PI 2: %inh MDAMB453 AB: BxPC3

(uM) 2.5 uM IC50 ( s M) IC50 (u M)

L46 <0.01 47.2.49 1.082 3.701

M46 <0.1 60.945

N46 <0.1 32.984 19.188 7.441

046 <0.1 12,231

P46 <0.1 -39.422 Example 282. Synthesis of 2-{4-(7-(cyclopropylamjno)-3-ib

a]pyriniidin-5-yl)piperazin-l-yl)nicotinonitrile

[00650] TerZ-butyl 5-chloro-3-formyl-6-methylpyrazolo[l,5-a]pyrimidin-7- yl(cyclopropyl)carbamate (60 mg, 0.17 mmoi) was mixed with 2-(piperazin-l-yl)nicotinonitrile (64 mg, 0.34 mmol) in /^'-propanol (1 niL). The reaction mixture was heated at 90°C in the microwave for 1 hour. The reaction was cooled to room temperature and concentrated under vacuum. The residue was dissolved in (1 : 1) TFA/DCM (4 mL) and stirred at room temperature for 1 hour. The reaction was evaporated to dryness, quenched with 3N NaOH, filtered, washed with water, and dried under vacuum The product, 2-(4-(7-(cyclQpropylamino)-3-formyl-6- rnethyipyrazolo[l ,5-a]pyrimidin-5-yl)piperazm-l -yl)nicotinonitrile, was further purified by preparative TLC using 5% acetone/dichioromethane as the eluent (40 mg, 58% yield). LCMS ( · i 403 )

Example 283, Synthesis of 2-(4-(7-(cvclopropylamino)-3-((2,5-dioxoimidazolidm-4- ylidene)methyl)-6-methylpyrazolo[ 1 ,5-a]pyrimidin-5-yl)piperazin- 1 -yl)nicotmonitrile

[00651 J Hydantoin (7.5 mg, 0.08 mmol) and 2-(4-(7-(cyclopropylamino)-3-formyl-6- methylpyrazolc 1,5-a]pyrimidin-5-yi)piperazin-l~yl)nicotinoni (15 mg, 0.04 mmol) were dissolved in ethanol (0.5 mL) along with piperidine (8 uL, 0.08 mmol). The reaction was heated at 80 °C for 1 hour in the microwave. The reaction was then cooled to r.i, diluted with water. and the precipitate was collected and washed with water, 1 : 1 ethanohwater, then ethanol. The yellow solid w^ras dried in vacuo to give 2-(4-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin- 4-yUdene)methyl)-6-methylpyrazolo[ 1 ,5-a]pyrirmdin~5~yl)piperazin- 1 -yljnicoti omtrile (3 mg, 17% yield). LCMS { M · 1 485 }

Example 284. Synthesis of 2-{4-(7-(cyclopropylaniino -3-((2,4-dioxothiazolidm-5- ylidenc)methyl)-6-methylpyrazolo [ 1 ,5 -a]pyrimidin-5 -yl)piperazin- 1 -yl)nico tinonitrile

[00652 J in a reaction flask, thiazolidine-2,4-dione (9 mg, 0.08 mmol) and 2-(4-(7- (cyclopropylamino)-3 -formyl-6-methylpyrazol o [ 1 , 5 -a]pyrimidin-5 -yl)piperazin- 1 - yl)nicotinonitrile (15 mg, 0.04 mmol) were dissolved in ethanol (0.5 mL) along with piperidine (8 uL, 0.08 mmol). The reaction was heated at 80 °C for 1 hour in the microwave. The reaction was then cooled to r,i, diluted with water, and the precipitate was collected and washed with water, 1 : 1 ethanohwater, then ethanol. The yellow solid was dried in vacuo to give 2-(4-(7- (cyclopropylamino)-3-((2,4-dioxotMazolidin-5-ylidene)methyl)-6-m [1 ,5- a]pyrimidin-5-y piperazin- 1 -yi)nicotinonitrile (10 mg, 54% yield). LCMS (M+l =502)

[00653] The following molecules were prepared using chemistries similar to synthesis in examples above. All compounds were characterized by LCMS. Table 55B shows the biological activities of the compounds lis ted in Table 55 A.

Table 55 A.

Table 55 B.

Example 285. Synthesis of tert-butyl 5~chloropyr3zolo[ 1. ,5-a]pyrimidin-7-y 1.{2- morpholmopropyl)carbamate

[00654] To the reaction flask, 5,7-dichloropyrazolo[l ,5-a]pyrimidine (3.2 g, 17 mmol) was added along with 2-morpholinopropan-l -amine (2.4 g, 17 mmol), triethylamine (2.3mL, 17 mmol), and acetonitrile (56 riiL), The reaction was stirred at 85°C for 12 hours then cooled to room temperature, diluted with water, filtered and washed with water. The intermediate, 5- chloro~N-(2~morphoiinopropy])pyrazolo[l ,5-a]p3'Timidin-7~amine, was dried under vacuum to provide 3.8 grams of an off-white solid (77% yield). LCMS (M+l= 296)

[00655] To 5-chloro-N-(2-morpholinopropyl)pyrazolo[l,5-a]pyrimidin-7-aniine (3.8 g, 13 mmol) in methylene chloride (50 mL) was added triethylamine (2.1 mL, 15 mmol),

dimethyiaminopyridine (200 mg, 1.6 mmol), and di-t-butyldicarbonate (3.3 g, 15 mmol). The mixture was stirred at room temperature for 10 hours. The reaction mixture was transferred to a separator}' funnel, washed IX with H?0, 2X with brine, dried over MgS0₄, filtered, and evaporated to dryness to provide an oily residue which solidified on standing. The product, tert- butyl 5-chloropyrazolo[l,5-a]pyrimidin-7-yl(2-morpholinopropyl}carbamate, was recovered as an off-white solid in 39% yield (5.1 mmol). LCMS ( M · i 396)

Example 286. Synthesis of N5 -(5 -chloro-2-fluorophenyl)-N7-(2- morpholinopropyl)pyrazolo[L5-a]pyrimidine-5,7-diamine

[00656] To tert-butyi 5-chloropyrazolo[ 1 ,5-a]pyrimidin~7-yi(2-mo^holinopropyi)carbamate (396 mg, 1 mmol), 5-chloro~2~fluoroaniime (145 uL, 1.2 mmol), and LiHMDS (2.2 mL, 2.2 mmol, 1M in THF) was added X-Phos (11 mg, 0.024 mmol) and

tris(dibenzylideneacetone)dipalladium(0) (18 mg, 0.02 mmol). The mixture was sealed and irradiated at 65 °C for 60 niiii in the microwave. The reaction was quenched with IN HCL (2 mL) and then neutralized with saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate and washed with saturated sodium chloride solution. The organic layer was collected, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude residue was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (2 mL). After stirring at room temperature for 1 hour, the solution was concentrated under a stream of nitrogen. The crude material was neutralized with saturated sodium bicarbonate solution then purified by silica gel chromatography (75% ethyl acetate/he anes) to yield the product, N5-(5- ch[oro-2-fluorophenyl)-N7-(2-niorpholin^ (84 mg, 21% yield). LCMS (M+ 1=405)

Example 287. Synthesis of 5-(5-chloro-2-fluorophenylamino )-7-(2- morpholmopropyiammo)pyrazolo[1 ,5-a]pyrimidine-3-carbaldehyde

[00657] In a reaction flask, N5-(5-chloro-2-fluorophenyl)-N7-(2- moiphoimopropyl)pyrazolo[l ,5-a]pyrimidine-5,7-dianiine (84 mg, 0.21mmol) was dissolved in DMF (0.9 mL) then phosphorous oxychloride (58 uL, 0.62 mmol) was added slowly at room temperature. The reaction mixture was allowed to stir at room temperature for 2 days then quenched by addition to 6N NaOH solution. The pH of the mixture was adjusted with 6N HC1 to pH = 7-9. The solid was recovered by filtration and washed with water. The product, 5-(5- chloro-2-fluorophenylammo)-7-(2-morph^

carbaidehyde, was purified by preparative TLC (5% acetone/dichloromethane) to yield 37 mg of the desired product (41% yield). LCMS (M+l=433)

Example 288. Synthesis of 5 -f f 5 -(5 -chloro-2-fluoropheny lamino)-7-(2- morpholinopropylarm^o)pyra^

[00658] Hydantoin (7 rng, 0.07 mmol) and 5-(5-chloro-2-fluorophenylarn.ino)-7-(2- morpholinopropylamino)pyrazolo[l ,5-a]pyrimidine-3-carbaldehyde (15 mg, 0.035 mmoi) were dissolved in ethanol (0.5 mL) along with piperidine (7 uL, 0,07 mmol), The reaction was heated at 80 °C for 1 hour in the microwave. The reaction was then cooled to r.t., diluted with water, and the precipitate was collected and washed with water, 1 : 1 ethanohwater, then ethanol. The yellow solid was dried in vacuo to give 5-((5-(5-chioro-2-tTuorophenylamino)-7-(2- moφhoiillo ro la.millo) yτa.zolo[l ,5- ] yrimidi -3-yl)mεthylelle)imidazoli (5 mg, 28% yield). LCMS ( M · 1 5 1 )

[00659] The following molecules were prepared using chemistries similar to synthesis in examples above. All compounds were characterized by LCMS. Table 56B shows the biological activities of the compounds listed in Table 56A.

Table 56A.

Table 56B.

Example 289, Synthesis of tert-butyl 4-{7-(cyclopropylamino)-3-fbmiylpyr3Z'

-5-yr)piperazine- 1 -carboxylate

[00660] To 5-chloro-7-(cyclopropylamino) pyrazolo [1, 5 -a] pyrimidine-3-carbaldehyde (500 mg, 2.1 1 mmo!) in dimethylformamide was added 1-Boc-piperazine (1.17g, 6.33 mmol), potassium carbonate (583mg, 4.21mmol) and di isopropyl ethylamine (0.41mL, 2.5 mmol). The mixture was heated to 80°C for overnight. Cooled the reaction mixture, added water and filtered the white precipitate to yield tert-butyl 4-(7-(cyclopropylamino)-3-formylpyrazolo [1, 5 -a] pyrimidin-5-yl) piperazine- 1 -carboxylate (655 mg, 80% yield). LCMS (M+l-387)

Example 290. S ynthesis of (Z)~ -(( 7-f cy clopropy j a.mino)~ -( pi perazin- 1 ~yl) yrazolo I^" L 5 - -3-yl)methylene)imidazo1idme-2,4-dione

[00661] The above product tert-butyl 4-(7-(cyclopropylamino)-3-formylpyrazolo [1, 5-a] pyrimidin-5-yl) piperazine- 1 -carboxylate (250 mg, 0.645 mmol) was dissolved in 2.0 raL ethanol, added hydantoin (129 mg, 1.288 mmol) and piperidine (127 ui). The reaction was heated to 80°C for three hours. Cooled the reaction mixture and yellow precipitate was filtered, washed with ethanol, dried to yield (Z)-tert-butyl 4-(7-(cyclopropylamino)-3-((2,5- dioxoirmdazolidin-4-ylidene)methyl)pyrazolo[l,5-

(264 mg, 87% yield). The above product was further dissolved inl : l mixture of DCM: TFA and stirred at room temperature for 30 minutes. Mixture was concentrated and dried to yield yellow solid of (Z)-5-((7-(cyclopropylamino)-5-(piperazin-l-yl) pyrazolo [1 , 5-a] pyrimidin-3-yl) methylene) iniidazolidine-2, 4-dione. LCMS (M+ 1=369)

Example 291. Synthesis of (Z)-5-((5-(4-(2-cyclopropylacetyl)piperazm- 1 -yl)-7-

[00662] To (Z)-5-((7-(cyclopropy[amino)-5-(piperazin-l-yl) pyrazolo [1, 5-a] pyrimidin-3-yl) methylene) imidazolidiiie-2, 4-dione (step b) (10 mg, 0.027 mmoi) in 50.0 uL NMP, was added HOBT (4.4 mg,0.032 mmol), cvclopropy 1 acetic acid (60 ul in 0.02M NMP solution), DIPEA (9.5 ul, 0.067 mmol) and EDC (7.8 mg, 0.040 mmol). The reaction mixture was stirred at room temperature for one hour. Diluted the reaction mixture with methanol and prepared by HPLC to yield (Z)-5-((5-(4-(2-cyclopropylacetyl)piperazin-l-yl)-7-(cyclopropylamino)pyrazolo[l,5- a]pyrimidin~3-yi)methylene)imidazoiidine-2,4-dione. LCMS (M+l ^:::451 )

[00663] The compounds listed in the following Table 57A were prepared according to the procedures as described above. Table 57B shows the biological activities of the compounds listed in Table 57A. Table 57A.

Table 57B.

Example 292. Synthesis of tert-buty 1 5 -azi do-3 -formylpyrazolo [ 1 ,5 -ajpyrimi din-7- ylfcyclopropyl Carbamate

[00664] To 5-chloro-7-(cyclopropylamino) pyrazolo [1 , 5 -a] pyrimidine~3-earbaldehyde (4.0g, 16.87 mmol) in diniethylformamide was added sodium azide (1.56 g, 23.9 mmol) and the reaction mixture was heated at 80°C for 8 hrs. Cooled the reaction mixture, added water and white precipitate filtered and dried to yield 5-azido-7-(cyclopropylamino) pyrazolo [1 , 5-a] pyrimidine-3-carbaldehyde (2.95 g, 75% yield). LCMS (M+ 1=244)

[00665 J The above product 5-azido-7-(cyclopropylamino) pyrazolo [1, 5-a] pyrimidine-3- carbaldehyde (1.09 g, 4,46 mmol) was subjected to hydrogenation using 10% wt palladium on carbon in ethanol. The reaction was stirred under hydrogen for 6 hours. The mixture was filtered through celite and sonicated with 1 : 1 mixture of ethyl acetate and hexane. The light yellow solid was filtered and dried to yield 5-amino-7-(cyclopropyiamino) pyrazolo [1, 5-a] pyrimidine-3-carbaldehyde as product 750 mg (85% yield). LCMS (M+ 1 =218)

Example 294, Synthesis of (Z^')-N-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidm-4- ylidene methyl.)pyrazolo[T,5-a^"lpyrimidin-5-yl -2-fluorobenzamide

[00666J To 5-amino-7-(cyclopropylaniino) pyrazolo [1 , 5-a] pyrimidine-3-carbaldehyde (step b) (30 mg, 0.138 mmol) in 1.0 mL tetrahydrofuran, stirring under nitrogen, was added 2-fluoro benzoyl chloride (33 ul, 0.275mmol) and DIPEA (28.8 ul). The reaction mixture was stirred at room temperature for one hour. The reaction was then partitioned between ethyl acetate and water, the organic layer was dried under sodium sulfate concentrated on high vaccum to yield N- (7-(cyclopropylamino)-3-formyIpyrazolo [1, 5-a] pyrimidin-5-yl)-2-fluorobenzamide. The crude product was further dissolved in 1 ,0 ml ethanol, added hydantom (41.2 mg, 0.41 1 mmol) and pipperdine (40,0 ul). The reaction was heated to 80°C for three hours. Cooled the reaction mixture and yellow precipitate was filtered, washed with ethanol to yield 10 mg (40% yield, two steps) (Z)-N-(7-(cyclopropylamino)-3-((2, 5-dioxoimidazolidin-4-ylidene) methyl) pyrazolo [1, 5-a] pyrimidm-5-yl)-2-fluoroberizamide. LCMS (M+ 1=422)

Example 295, Synthesis of (Z)-4-cyano-N-(7-(cyclopropylamino)-3-((2,5-dioxoimidazolidin-4- ylidene)methyi)pyrazolo[l ,5-a]pyrimidin-5-yl)benzamide

[00667] To 5-amino-7-(cyclopropylamino) pyrazolo [1, 5-a] pyrimidine-3-carbaldehyde (step b) (30 mg, 0.138 mmol) in 1 .0 ml. acetonitrile was added HATU (104 mg, 0.273 mmol), 3- cyano benzoic acid (30 mg, 0.203 mmol) and DIPEA (48,0 ul), The reaction mixture was heated to 80°C for five hours. Cooled the reaction mixture and light yellow precipitate was filtered, washed with acetonitrile to yield 4-cyano-N-(7-(cyclopropylamino)-3-formylpyrazolo [1, 5-a] pyrimidin-5-yi) benzamide. The crude product was further dissolved in 1.0 mL ethanol, added hydantoin (10 mg, 0.01 mmol) and pipperdine (9.5 ul). The reaction was heated to 80°C for three hours. Cooled the reaction mixture and precipitate was filtered, washed with ethanol to yield 7 mg (40% yield, two steps) (Z)-4-cyano-N-(7-(cyclopropyiamino)-3-((2,5- dioxoimidazo[idin-4-ylidene)methyl)pyrazolo[l,5-a]pyrimidin-5-yl)benzam LCMS

(M 1 429)

[00668] The following compounds were prepared using chemistries similar to synthesis in examples above with the corresponding acids, chloroformates, or isocyanates. All compounds were characterized by LCMS. Table 58B shows the biological activities of the compounds listed in Table 58 A. Table 58 A.

412

413

414

Table 58B.

[00669] The following compounds were prepared using chemistries described in the present disclosure. Table 59B shows the biological activities of the compounds listed in Table 59A,

Table 59A.

421

[00670] The following compounds in Table 60 can be prepared using chemistries described in the present disclosure: Table 60,

[00671 J Compound 3 were prepared by reaction compound 1 with boronic acid 2 using Suzuki coupling reaction conditions as shown below (Scheme 2),

[00672] The following compounds can be prep in Table 61 can be prepared by using chemistr}_' described in Scheme 2.

Table 61.

[00673] Compound 5 (as shown in Scheme 3 below) were prepared from compound 4 using chemistries described in patent application US2004/0019058. Compound 5 can be converted to molecule 6 using intermediates and chemistries described above, Deproteciion of 6 using reagents such as Ν Ν-dimethylbarbituric acid and a palladium catalyst can lead to 7.

[00674 J Compound 7 (as shown in Scheme 4 below) can be converted to molecules 8, 9, 10, 11 and 12 using chemistries known to a person skilled in the art.

[00675] The following compounds in Table 62 can be prepared using chemistries described in the present disclosure: Table 62,

[00676] Compoimd 3 were prepared by reaction compound 1 with boronic acid 2 using Suzuki coupling reaction conditions (as shown in Scheme 5 below).

1

Scheme s

)677] The following compounds in Table 63 can be prepared using chemistry described on

Table 63.

[00678] Compound 2 were prepared by reaction compound 1 using reductive animation c

00679] The following compounds in Table 64 can be prepared using chemistry described on

Table 64,

Biological Test Methods:

Etiological Example A

CK2 Assay Method

[00680] Modulator}_' activity of compounds described herein was assessed in vitro in cell-free CK2 assays by the following method.

[00681] In a final reaction volume of 50 μ,ί, CK2 ααββ (4 ng, 8.5 mil) was incubated with various concentrations of test compounds in DMSO (1 ul, 2% by volume), 20 mM MOPS pH 7.2, 10 mM EGTA, 0.15 M NaCL 10 mM DTT, 0.002% Brij-35, 200 μΜ RRRDDDSDDD, 10 mM MgAcetate, ATP 15 uM and 0.33% (by volume) ([γ-33Ρ]ΑΤΡ: Stock lmCi/ΙΟΟμΙ;

3000Ci/'mmol (Perkin Elmer)). Reactions were maintained for 40 min at 23 °C. The reactions were quenched with 100 ul of 0.75% Phosphoric acid, then transferred to and filtered through a Phosphoceilulose filter plate (Miliipore, MSPH~N6B~50). After washing each well 4 times with 0.75% Phosphoric acid, scintillation fluid (20 uL) was added to each well and the residual radioactivity was measured using a Wallac luminescence counter.

Biological Example B

PIM-1 Assay Method

[00682] The following procedure was used to assay the PIM-1 kinase activity of compounds of the in v ention. Other methods for assaying PIM-1 and other PIM kinases, as well as methods to assay for activity against the various kinases for the kinase panel mentioned in Figures 1 and 2, are known in the art.

[00683] In a final reaction volume of 50 ul, recombinant PIM-1 (1 ng) was incubated with 12 mM MOPS pH 7.0, 0.4 mM EOT A, glycerol 1 %, brij 35 0,002 %, 2-mercaptoethanol 0.02 %, BSA 0.2 mg/ml, 100 uM KRNRTLT , 10 mM MgAcetate, 15 uM ATP, [γ-³³Ρ-ΑΤΡ] (specific activity approx. 500 cpni/pmol), DMSO 4%» and test inhibitor compound at the required concentration. The reaction was initiated by the addition of the Magnesium ATP mixture. After 40 min incubation at 23°C, the reactions were quenched by the addition of 100 ul 0.75% Phosphoric acid, and the labeled peptide collected by filtration through a

phosphoceilulose filter plate. The plate was washed 4 times with 0.075% phosphoric acid (100 ul per well) and then, after the addition of scintillation fluid (20 ul per well), the counts were measured by a scintillation counter. Biological Example C

Pi fvj - 2 Assay Method

[00684] Test compounds dissolved and diluted in DMSO (2 μΐ) were added to a reaction mixture comprising 10 μΐ of 5X Reaction Buffer (40mM MOPS pH 7.0, 5mM EDTA), 10 μΐ of recombinant human P1M2 solution (4 ng P1M-2 dissolved in dilution buffer (20 m MOPS pH 7.0; EDTA 1 mM; 5% Glycerol; 0.01% Brij 35; 0.1 %; 0.1 % 2-mercaptoethanoi; 1 rag/ml BSA)) and 8 ul of water. Reactions were initiated by the addition of 10 ul of ATP Solution (49% (1 mM MgCi₂; 75 uM ATP) 1 % ([γ-³³Ρ]ΑΤΡ: Stock lmCi/Ί ΟΟμΙ; 3000Ci/mmol (Perkin Elmer)) and 10 ul of substrate peptide solution (RSRSSYPAGT, dissolved in water at a concentration of 1 mM), Reactions were maintained for 10 rain at 30°C, The reactions were quenched with 100 ul of 0.75% Phosphoric acid, then transferred to and fiitrered through a Phosphocellulose filter plate (Millipore, MSPH-N6B-50). After washing each well 4 times with 0.75% Phosphoric acid, scintillation fluid (20 uL) was added to each well and the residual radioactivity was measured using a Wallac luminescence counter, Biological Example D

Cell Proliferation Modulatory Activity

[00685] A representative cell-proliferation assay protocol using Alamar Blue dye (stored at 4°C, use 20 ul per well) is described hereafter.

96-well plate setup and compound treatment

[00686] a. Split and trypsinize cells.

[00687] b. Count cells using hemocytometer.

[00688] c. Plate 4,000-5,000 cells per well in 100 μΐ of medium and seed into a 96-well plate according to the following plate layout. Add cell culture medium only to wells B 10 to B 12. Wells Bl to B9 have cells but no compound added.

1 2 3 4 5 6 7 8 9 10 1 1 12

A EMPTY

Medium

B NO COMPOUND ADDED

Only

C ΙΟηΜ ΙΟΟ^ηΜ luM l OuM Control

D Ι ΟηΜ ! OOnM l uM lOuM Com l

E lOiiM Ι ΟΟ^ηΜ luM lOuM Comp2

F ΙΟ^ηΜ lOOnM luM l OuM Comp3

G ΙΟηΜ !OOnM luM l OuM Comp4

H EMPTY

[00689 J d. Add 100 μΐ of 2X drug dilution to each well in a concentration shown in the plate layout above. At the same time, add 100 μΐ of media into the control wells (wells B10 to B12), Total volume is 200 μΐ /well.

[00690] e. Incubate four (4) days at 37°C, 5% CO_?, in a humidified incubator.

[00691] f. Add 20μ1 Alamar Blue reagent to each well.

[00692] g. Incubate for four (4) hours at 37°C, 5% CO? in a humidified incubator.

[00693] h. Record fluorescence at an excitation wavelength of 544nm and emission wavelength of 590nm using a microplate reader.

[00694] In the assays, cells are cultured with a test compound for approximately four days, the dye is then added to the cells and fluorescence of non-reduced dye is detected after approximately four hours. Different types of cells can be utilized in the assays (e.g., HCT-1 16 human colorectal carcinoma cells, PC-3 human prostatic cancer cells, M DA-MB231 human breast cancer cells, K-562 human chronic myelogenous leukemia (CML) cells, MiaPaca human pancreatic carcinoma cells, MV-4 human acute myeloid leukemia cells, and BxPC3 human pancreatic adenocarcinoma cells).

[00695] Activity of compounds of the present invention tested in these in vitro and cellular assays are summarized in Tables 1 A and 2A below. The compounds listed in Tables 1 A and 2A are the Examples and species as described above. Table Al. Bioactivity Data for Some Compounds of Fonnuia II and Formula IF.

Table A2, Bioactivity Data for Some Compounds of Formula II and Formula IF

[00696] Citation of the above patents, patent applications, publ ications and documents is not an admission that any of the foregoing is pertinent prior art, nor does it constitute any admission as to the contents or date of these publications or documents. Furthermore, the contents of the patents, patent applications, publications and documents cited herein are incorporated by reference in their entirety for ail purposes to the same extent as each and everyone of them is incorporated by references specifically.

[00697] Modifications may be made to the foregoing without departing from the basic aspects of the invention. Although the in vention has been described in substantial detail with reference to one or more specific embodiments, those of ordinary skill in the art will recognize that changes may be made to the embodiments specifically disclosed in this application, and yet these m odifications and improvements are within the scope and spirit of the invention. The invention illustrati vely described herein suitably may be practiced in the absence of any element(s) not specifically disclosed herein. Thus, for example, in each instance herein any of the terms "comprising", "consisting essentially of", and "consisting of may be replaced with either of the other two terms. Thus, the terms and expressions which have been employed are used as term s of description and not of limitation, equivalents of the features shown and described, or portions thereof, are not excluded, and it is recognized that various modifications are possible within the scope of the invention.

Claims

We claim:

1 , A compound of Formula ( II ) or (IF):

( II) (IF)

or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof;

wherein:

Z^J and Z⁴ each independently represent N or CR⁵, or CH:

each R^J is independently selected from halo, -CN, ~R, -OR, ~S(0)_nR, -COOR, -CONR₂, and -NR₂,

wherein each R is independently selected from H and optionally substituted C 1 -C4 alkyl, or alternatively, the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 or 6 membered heterocyclic ring that optionally contains one or more additional heteroatom selected from N, O and S as a ring member;

R², R³ and R⁴ are each independently selected from H and optionally substituted

CI -CI O alkyl;

X represents O, S, or NR²;

Y is O or S or NR.¹⁰;

where R'° is selected from H, CN, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted C.1 -C4 alkoxy, and -NR^?R⁸,

Z is O or S;

L is a bond, -CR⁷==CR⁸~, -C≡C-, -NR⁷-, -0-, -S(0)_N-, -(CR⁷R^s)_ai-, ~(CR⁷R⁸)_m~NR⁷~, - (CR⁷R⁸)_OT-0-, or -Ci R ^'R V--S( }_.,·-: W is optionally substituted CI -CIO aikyl optionally substituted CI -CIO heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, -NR^?R⁸, -OR', -S(0)_nR^?, - CONR'R , optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted C2-C10 aikenyl, optionally substituted C2-C10 alkynyl, or -CR'R⁸R⁹;

where each R' and R and R is independently selected from H, optionally substituted CI -CIO alk L optionally substituted heteroalkyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted carbocyclyialkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylaikyi, optionally substituted heteroaryl, and optionally substituted heteroaryialkyl;

or R⁸ and R^'' , taken together with the carbon atom to which they are attached, form O (oxo) or =N-OR' or =N-CN;

or R' and R⁸, taken together on a single carbon atom or on adjacent connected carbon atoms of (CR⁷R^s)_m whether alone or as part of another group, form a 3 to 8 membered carbocyclic ring or heterocyclic ring;

or R and R⁸, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5 to 10 membered heterocyclic or heteroaryl ring that optionally contains one or more additional heteroatom selected from N, O and S as a ring member;

provided that no more than one of or R and R⁸ in -NR^?R⁸ is selected from the group consisting of alkoxy, alkylamino, dialkylamino and heterocyclyl;

each n is independently is 0, 1 or 2;

each m is independently 1 , 2, 3 or 4; and

R^iA and R'^b are each independently selected from H, optionally substituted CI -C IO alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyl, optionally substituted carbocyclyl, optionally substituted carbocyclyialkyl, optionally substituted heterocyclylalkyl, optionally substituted aryl, optionally substituted arylaikyi, optionally substituted heteroaryl, or optionally substituted heteroaryialkyl;

or R^IA and R^lB, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5- to 8-mernbered monocyclic or 5- to 10-mem.bered bicyclic heteroaryl or heterocyclic ring containing up to two additional heteroatoms selected from N, O and S as ring members.

2. The compound of claim 1 , which is a compound of Formula (I I). The compound of claim 1 , which is a compound of Formula (IF).

4, The compound of any one of claims 1 to 3, wherein Z

5. The compound of any one of claims 1 to 4, wherein Z⁺ is CH.

6 The compound of any one of claims 1 to 3, wherein Z^" is CH.

7. The compound of any one of claims 1 to 4 or 6, wherein Z⁴ is N.

8, The compound of any one of claims 1 to 7, wherein R' and R⁴ are both H,

9 The compound of any one of claims 1 to 8, wherem R" is H, -CH₃, halo, OCH₃, or CF₃.

10. The compound of any one of claims 1 to 9, wherein Y is O or S.

1 1. The compound of any one of claims 1 to 9, wherein Z is O.

12. The compound of any one of claims 1 to 1 1 , wherein X is NFL

13. The compound of any one of claims 1 to 1 1 , wherein X is O or S.

14. The compound of any one of claims 1 to 13, wherein

the optionally substituted carbocyclyl is an optionally substituted C3-C8 cyeloaikyi; the optionally substituted carbocyclylaikyi is an optionally substituted C4-C10 cycloalkySaikyl; and

the optionally substituted heteroaikyi is an optionally substituted C1-C6 aikoxy, optionally substituted C1-C6 alkylamino, or optionally substituted C1-C6 dialkylamino.

15, The compound of any one of claims 1 to 14, wherein L is a bond or NFL

16. The compound of any one of claims 1 to 15, wherein W is optionally substituted phenyl, optionally substituted phenylalkyl, optionally substituted cycloalkylalkyl, or optionally substituted heterocyclyl.

17. The compound of any one of claims 1 to 14, wherein

-L-M is Ni III . -OR⁷, or -S(0)_nR⁷;

n is 0. 1, or 2; and

R' is optionally substituted CI -CIO alkyi, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted arylalkyl, optionally substituted he eroarylalkyl, optionally substituted carbocyclyl, optionally substituted

heterocyclyl, optionally substituted carbocycK alkyl, or optionally substituted heterocyclylalkyl.

18. The compound of any one of claims 1 to 14, wherein

-L-M is -NR⁷R⁸: and

I ^' and R , taken together with the nitrogen atom to which they are attached, form an optionally substituted hetercyclyi which optionally contains one or more additional heteroatom as ring members.

19. The compound of any one of claims 1 to 14, wherein

-L-M is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocycyl, or optionally substituted heterocyclyl.

20. The compound of any of claims 1 to 20, wherein R¹" is H and R^1B is optionally substituted CI -C10 alkyi, optionally substituted heterocyclyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted heterocyclylalkyl, optionally substituted arylalkyl, or an optionally substituted 5-6 membered aryl ring containing up to two heteroatoms as ring members.

21. The compound of claim 1 , which has the structural Formula (Ila) or (Ha'):

(Ila) (Ila') wherein

7 and Z⁴ each independently represent N or CR\ or CH;

where each R³ is independently selected from halo, -CN, -R, -OR, -S(Q)_r.R, -COOR, - C0\ R\ and -NR₂,

wherein each R is independently selected from H and optionally substituted C1 -C4 alkyl, or the two R groups, taken together with the nitrogen atom to which they are attached, form an optionally substituted 5- or 6-membered heterocyclic ring which contains one or more additional heteroatom selected from N, O and S as a ring member;

R⁴ is Π. CH₃ or CF₃;

X is O, S or M i :

Y is O or S;

R^{¾ B} is selected from H, optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted heterocyclyi, optionally substituted eycioalkyl, optionally substituted cyeloalkylalkyl, optionally substituted heterocyclylalkyl, optionally substituted arylalkyl, or an optionally substituted heteroaryl;

L is a bond, -NR⁷-, -0-, S( ( ) },··. (CR⁷R⁸)_m, or -(CR⁷R⁸)_m-NR⁷-;

m is 1 , 2, 3, or 4;

n is 0, 1 , or 2;

W is selected from optionally substituted aryl, optionally substituted heteroaryl, and - R^{7 8},

where each R and R^l is independently selected from H, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 alkyianiino, optionally substituted C1-C6 dialkylamino, optionally substituted heterocyclyl, optionally substituted CI -CIO alky], optionally substituted C3-C8 cycloalkyl, optionally substituted C4-C 10 cycloalkylalkvl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;

and R⁷ and R⁸, taken together on a single carbon atom or on adjacent connected carbon atoms of (CR R⁸)_m whether alone or as part of another group, form a 3- to 8-membered ring that contains one or more heteroatoms as ring members;

or R' and R' , taken together with the nitrogen atom to which they are attached, form an optionally substituted 5- to 10-membered heterocyclic or heteroaryl ring system that optionally contains an additional heteroatom selected from N, O and S as a ring member; and

provided that no more than one of or R' and R in -NR 'R is selected from the group consisting of alkoxy, alkylamino, dialkylamino and heterocyclyl,

22. The compound of claim 21, wherein R², R⁴ and R⁵ are each H.

23, The compound of claim 21 or 22, wherein X is NH and Y is O.

24. The compound of any one of claims 21 to 23, wherein R^1B is C3-C8 cycloalkyl or C4-C8 cycloalkylalkvl ,

25, The compound of any one of claims 21 to 24, wherein Z³ is N or CH.

26. The compound of any one of claims 21 to 25, wherein Z⁴ is N or CH,

27, The compound of any one of claims 21 to 26, wherein Z³ and Z⁴ are both N or Z³ and Z⁴ are both CH.

28. The compound of any one of claims 21 to 27, wherein L is NH.

29, The compound of any one of claims 21 to 28, wherein W is optionally substituted phenyl or optionally substituted thienyl.

30. The compound of any one of claims 21 to 29, wherein W is optionally substituted phenylalkvl, optionally substituted cyeioalkylalkyi, or optionally substituted heterocvclyl.

31. The compound of any one of claims 21 to 27, wherein

-L-M is -NHR⁷, -OR⁷, or -S(Q)„R⁷;

n is 0, 1 , or 2; and

R' is optionally substituted CI -CIO alkyl, optionally substituted heteroalkyl, optionally substituted aryi optionally substituted heteroaryi optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted carbocyclvl, optionally substituted

heterocvclyl, optionally substituted carbocyclyialkyl, or optionally substituted heterocyclylalkyl.

32. The compound of any one of claims 21 to 27, wherein

-L-M is \ R^"R^':; and

R' and R⁵, taken together with the nitrogen atom to which they are attached, form an optionally substituted hetereyelyl which optionally contains one or more additional heteroatoni as ring members.

33. The compound of any one of claims 21 to 27, wherein

-L-M is optionally substituted aryl, optionally substituted heteroaryi, optionally substituted carbocycyl, or optionally substituted heterocvclyl.

34. The com ound of claim 21 , which has the structural Formula (II-Th) or (II-Th'):

or

(II-Th) (II-Th')

wherein R ^{i ::} is selected from H, halo, optionally substituted C1~C6 aikyi. CN, S(Q)o-₂R, -S0₂NR₂, COOR, CONR₂, and C(0)R,

where each R is independently 1-1, halo, CN, or an optionally substituted member selected from the group consisting of C1 -C6 alkyl, C1 -C6 a!koxy, C1-C6 aikyiamino, di(Cl- C6)alkylaniino, C3-C8 cycloalkyL C4-C1G cycloalkylalkyL C5-C8 lieterocyclyl, C6-C10 heterocyclylalkyl, aryl, aryialkyl, C5-C6 heteroalkyi, and C6-C10 heteroaikyialkyl; and

two R on the same atom or adjacent connected atoms can form an optionally substituted heterocyclic ring that can contain an additional heteroatom selected from N, O and S as a ring member;

or a pharmaceutically acceptable salt thereof.

35. The compound of claim 34, wherein Y is O, and X is NH or S.

36. The compound of claim 34 or 35, wherein and R⁴ are each H.

37. The compound of any of claims 34 to 36, wherein R^TH is CONR₂.

38. The compound of any of claims 21 to 37, wherein R^3b is cyclopropyl or

cyeiopropylrnethyl.

The compound of claim 1, which has the structural Formula (lib) or (lib')

(lib) (lib') wherein

R^~" and R⁴ are independently H, CH₃ or CF₃;

Z⁴ is N or CH: is -NR^SAR , M IR . OR . or SiOj .R :

n is 0, 1, or 2; and

R' is optionally substituted C1-C10 alkyl, optionally substituted heteroalkyl, optionally substituted a d, optionally substituted lieteroaryl, optionally substituted arylalkyl, optionally substituted heteroarylalkyl, optionally substituted carbocyclyl, optionally substituted

heterocyclyl, optionally substituted carbocyclylaikyi, or optionally substituted heterocyelylalkyl optionally substituted aryl, optionally substituted lieteroaryl, optionally substituted carbocycyl, or optionally substituted heterocyclyl; or

R' and R ' , taken together with the nitrogen atom to which they are attached, form an optionally substituted hetercyclyl which optionally contains one or more additional heteroatom as ring members,

40. The compound of claim 1, which has the structural Formula (He):

wherein.

X is O, S, or R ;

R³ is -(CH₂)~X^C;

X^c is hydroxy! or a group having structural formula (a), (b), (c), or (d):

L¹ and L² are each independently a covalent bond, -0-, or 3a R ^la and FT^a are each independently hydrogen, alkyl, heteroalkyl, heteroaryl, heterocyciyl, alkenyl, alkynyl, arylaikyi, heteroaryialkyl, heterocycivlalkyl, -alkylene-C(0)-0-R^4a, or -alkylene-0~C(0)-0-R^4a; and

R^3i! and R^4a are each independently hydrogen, alkyl, heteroalkyl, cyclyialkyl,

heterocyciyl, aryl, heteroai l, alkenyl, alkynyl, arylaikyi, heterocyclylalkyl, or heteroaryialkyl;

L^"' is a covalent bond or alkylene;

Y is OR^5a, NR^5aR^6a, or C(0)OR^7a, provided that when Y is C(0)OR^7a, then L³ is not a covalent bond; and

R^5a, R^oa, and R'^a are each independently hydrogen, alkyl, arylaikyi, aryl, heteroalkyl, alkylheteroaryl, heterocyciyl, or heteroaryl; or alternatively, R⁵³ and R^6a, taken together with the nitrogen atom to which they are attached, form a hetercyclyl ring optionally containing one o rmore additional heteroatom independently selected from N, O, and S.

41. The compound of claim 40, wherein

X is NR²;

R³ is ^~(CH₂)-X^C;

X^1" is hydroxy! or a group havin stnictura! formula (b):

42. The compound of claim 40 or 41 , wherein R' and R^" are hydrogen.

43. The compound of any of claims 40 to 42, wherein R^{5 B} is an optionally substituted C I - CIO alkyl, cycloaikyi, or cycloalkylalkyl.

44. The compound of any one of claims 40 to 43, wherein -L-W is -OR^' or -N R .

45. The compound of claim 44, wherein R' is optionally substituted aryl or optionally

8

substituted heteroaryl; and R is H.

46. The compound of claim 45, wherein R⁸ is optionally substituted phenyl.

47. The compound of any one of claims 41 to 46, wherein L^J is a covalent bond; and Y is OR^5a or NR^5aR^6a.

48. The compound of claim 1, which is selected from the species disclosed in the

specification.

49. A pharmaceutical composition comprising

a compound of any one of claims 1 to 48; and

at least one pharmaceutically acceptable excipient.

50. The pharmaceutical composition of claim 49, further comprising one or more additional therapeutic agent,

51. The pharmaceutical composition of claim 50, wherein the one or more additional therapeutic agent is an anticancer agent.

52. A method for modulating casein kinase 2 activity and/or Pirn kinase activity in a ceil comprising contacting the cell with a compound of any one of claims 1 to 48, or a

pharmaceutically acceptable salt, solvate, and/or prodrug thereof.

53. A method of treating a condition or disease associated with casein kinase 2 activity and/or Pim kinase activity in a patient comprising administering to the patient a therapeutically effective amount of the compound of any one of claims 1 to 48, or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof.

54. The method of claim 53, wherein the condition or disease is selected from a group consisting of a cancer, a vascular disorder, a inflammation, a pathogenic infection, a

immunological disorder, a neurodegenerative disorder, and a combination thereof.

55. The method of claim 54, wherein the cancer is of the colorectum, breast, lung, liver, pancreas, lymph node, colon, prostate, brain, head and neck, skin, liver, kidney, blood and heart.

56. The method of claim 53, comprising administering to the patient the compound of any one of claims 1 to 48, or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof, in combination with one or more additional therapeutic agent.

57. The method of claim 56, wherein the one or more additional therapeutic agent is an anticancer agent.

58. A method for inhibiting cell proliferation, which comprises contacting cells with the compound of any one of claims 1 to 48, or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof, in an amount effective to inhibit proliferation of the cells.

59. The method of claim 58, wherein the cells are in a cancer cell line or in a tumor in a subject.

60. The method of claim 59, wherein the cancer cell line is a breast cancer, prostate cancer, pancreatic cancer, lung cancer, hematopoietic cancer, colorectal cancer, skin cancer, ovary cancer cell line.

61. A method for inhibiting angiogenesis in a subject, which comprises administering to the subject the compound of any one of claims 1 to 48, or a pharmaceutically acceptable salt, solvate, and/or prodrug thereof, in an amount effective to inhibit the angiogenesis.