RU2691754C1 - DERIVATIVE OF ZINC METAL COMPLEX CHLORINE-e6 AND USE THEREOF - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to a chlorine-e₆ zinc metal complex derivative general formula:

.

What is also presented is using the derivative as an agent for photodynamic therapy.

EFFECT: invention increases homogeneity, improves water solubility, increases selectivity of tumor accumulation compared to healthy tissues, reduces systemic toxicity.

6 cl, 18 dwg, 2 tbl

Description

The proposed group of inventions relates to the field of biomedicine, relates to a derivative of the zinc metal complex chlorin-e ₆ , which can be used as an agent used in photodynamic therapy (PDT) for the diagnosis and treatment of cancer.

Oncological diseases are currently one of the most acute medical and social problems of mankind. One of the main non-surgical methods of treating various forms of tumor diseases is the method of photodynamic therapy. However, the existing anticancer drugs have a number of serious drawbacks, including low accumulation selectivity in tumor tissues and insufficient water solubility, which leads to an increase in toxicity and a decrease in the effectiveness of PDT. Thus, the creation of new PDT agents with improved pharmacokinetic parameters is a pressing public health task. To date, a number of compounds known to exhibit light antitumor toxicity with respect to tumor cells. The therapeutic effect of these photosensitizer molecules is due to the generation of reactive oxygen species and radicals under the action of light radiation, which leads to the activation of various death mechanisms of tumor tissues [P. Mroz, A. Yaroslavsky, G.B. Kharkwal, M.R. Hamblin, Cell Death Pathways in Photodynamic Therapy of Cancer, Cancers 3 (2011) 2516-2539].

A large number of photoactive compounds based on chlorine photosensitizers are known. Some of them are successfully used in clinical practice.

For example, PHOTOLON (RU 2152790 C1, cl. A61K 31/79, G01N 33/52, publ. 07.20.2000), the main active ingredient of which is chlorin-e ₆ , which is 18-carboxy-20- (carboxymethyl) -8-ethenyl-13-ethyl-2,3-dihydro-3,7,12,17-tetramethyl-21H, 23H-porphin-2-propionic acid. In the dosage form, chlorin-e _{6 is} contained in the form of trisodium salt together with low molecular weight medical polyvinylpyrrolidone.

Another drug based on a photosensitizer is RADACHLORIN (RU 2183956 C1, class A61K 31/409, A61P 35/00, publ. 06/27/2002), which is a mixture of chlorin-e ₆ , purpurin-5, and chlorin-p6.

Another well-known drug PHOTODITAZIN (RU 2144538 C1, cl. C07D 487/22, A61K 31/40, A61P 35/00, publ. 01/20/1998) is the bis-N-methyl-D-glucamine salt chlorin-e ₆ in the form of an aqueous solution containing polyvinylpyrrolidone. Available in the form of a concentrate and gel penetrator.

In addition, an effective PDT agent is the Japanese drug NPe ₆ (Talaporfin) (US 2008254570 A1, CL H01L 21/00, published on 10/16/2008), which is the sodium salt of N-aspartyl-chlorin-e ₆ .

One of the most important characteristics of the photosensitizer is the uniformity of the composition. This is due to the fact that chlorine-e ₆ -based compounds have the highest quantum yields of singlet oxygen, while impurity components are often less active.

PHOTOLON, PHOTODITAZIN, RADACHLORIN and NPe ₆ are currently being actively used to treat various oncological diseases, but they have several disadvantages, such as composition heterogeneity and low selectivity of accumulation in tumor tissue, which significantly reduces their overall effectiveness. Preparations PHOTOLON, PHOTODITAZIN, RADACHLORIN and NPe ₆ are aqueous solutions consisting of the main active ingredient based on chlorin-e ₆ , as well as a number of different additives. The amount and composition of additives can vary within different limits. The most chemically homogeneous agent is PHOTODITAZIN, however, it has a limited shelf life (about 1 year) and only in the refrigerating chamber. Therefore, the creation of a homogeneous in composition PDT agent is a promising direction in the creation of new drugs. Also, PHOTOLON, PHOTODITAZIN, RADACHLORIN and NPe ₆ preparations are 2nd generation photosensitizing agents, the main disadvantage of which is the low distribution selectivity between healthy and tumor tissues [Materials 2015, 8 (7), 4421-4456].

To increase the selectivity of photosensitizer accumulation in tumors, photoactive hybrid conjugates with targeted (directing) fragments are created.

For example, it has been shown that molecules that contain targeted molecules, such as antibodies, sialic acids, biotin, some peptides selectively binding to tumor cells, can significantly reduce toxicity and increase the activity of the drug based on them by selectively binding to oncoexpressible receptors. on the surface of cells [J. Pharm. Sci. 103 (2014) 71-77; J. Mater. Chem. B 4 (2016) 6758-6772].

A preparation is known based on the chlorin-e ₆ conjugate bound to transferrin protein, whose receptors are over-expressed by tumor tissues (US 20020137901 A1, cl. C07K 14/79, A61K 38/00, A61K 41/00, C07K 14/435, publ. 26.09 .2002).

Also known is a drug based on chlorin-e ₆ conjugate associated with folic acid, which allows delivery of the photoactive part directly to tumor cells by binding to folate receptors on their surface (US 2012059018 A1, A61K 31/519, A61P 35/00, C07D 487/22, published on March 8, 2012).

Also developed a derivative of chlorin-e ₆ , covalently linked to oligonucleotides, showing high efficiency of binding to malignant tumors (US 20060105974 A1, class A61K 48/00, A61K 38/00, A61K 41/00, A61K 47/48, С07Н 21 / 00, C07N 21/04, C12N 15/113, published on 05/18/2006).

Among the existing PDT conjugates, the closest to the proposed invention is the conjugate of the purpurinimide metal complex with mono- and disaccharides (US 2002198157 A1, class A61P 35/00, С07Н 15/26, С07Н 23/00, publ. 12.26.2002). These conjugates consist of a photoactive core based on the metal complex of purpurinimide, associated with carbohydrate fragments (Fig. 1). The conjugates described above were developed at the Roswell Park Cancer Institute in the group of Dr. Pandey. The structures of these molecules were selected on the basis of focusing on the therapeutic target in the form of Galectin-3, expressed by some tumor cells. A number of compounds were synthesized with various carbohydrate molecules linked to purpurinimide; several types of linkers were also chosen for their connection. It has been shown that biological activity strongly depends on the nature of the carbohydrate and the type of linker. For example, molecules that do not contain carbohydrate residues demonstrate a decrease in activity compared to analogues with galactose and lactose.

Compound 4 - ((4'-bromo-2'-fluorophenyl) amino) -6-methoxy-quinazolin-7-ol is an active part of the ligand receptors for epidermal growth factor (EGFR) and vascular endothelium (VEGFR), as well as an effective tyrosine kinase inhibitor, which makes it a potentially convenient delivery agent and independent chemotherapeutic agents [J. Med. Chem. 2012, 55, 10797; Clin. Ther. 2011, 33, 315; Clin. Ther. 2012, 34, 221].

The task of the invention is the creation of a new derivative of the zinc metal complex chlorin-e ₆ , linked through chemical bonds with 4 - ((4'-bromo-2'-fluorophenyl) amino) -6-methoxy-quinazolin-7-ol, which improves the biodistribution of the photo-sensitizer during by binding to EGFR / VEGFR receptors on the surface of tumor cells, as well as its use as an agent for photodynamic therapy.

The technical result from the use of the proposed group of inventions is to increase homogeneity, improve water solubility, increase the selectivity of the accumulation of tumor tissues compared with healthy, reducing systemic toxicity.

The task is achieved by the fact that the derivative of the zinc metal complex chlorin-e ₆ has the general formula:

производное представляет собой бромид цинкового комплекса 13-(диметилэтиламмонийэтил)амид-17(3)-(1ʺ-дезокси-β-D-галактопиранозил-1ʺ-(1',2',3'-триазол-4'-ил)-метил)амидхлорина-е₆, общей формулы:at

the derivative is a zinc complex bromide 13- (dimethylethylammonium ethyl) amide-17 (3) - (1ʺ-deoxy-β-D-galactopyranosyl-1ʺ- (1 ', 2', 3'-triazol-4'-yl) -methyl ) amidchlorin-e ₆ , of the general formula:

производное представляет собой бромид цинкового комплекса 13-(диметилэтиламмонийэтил)амид-17(3)-(1ʺ-дезокси-β-D-глюкопиранозил-1ʺ-(1',2',3'-триазол-4'-ил)-метил)амидхлорина-е₆ общей формулы:at

the derivative is a zinc complex bromide 13- (dimethylethylammonium ethyl) amide-17 (3) - (1ʺ-deoxy-β-D-glucopyranosyl-1ʺ (1 ', 2', 3'-triazol-4'-yl) -methyl ) amidchlorin-e _{6 of the} general formula:

производное представляет собой цинковый комплекс 13-(диметиламиноэтил)амид-17(3)-(1ʺ-дезокси-β-мальтозил-1ʺ-(1',2',3'-триазол-4'-ил)-метил)амидхлорина-е₆ общей формулы:at

the derivative is a zinc complex of 13- (dimethylaminoethyl) amide-17 (3) - (1ʺ-deoxy-β-maltosyl-1ʺ- (1 ', 2', 3'-triazol-4'-yl) methyl) amidchlorin- e _{6 of the} general formula:

представляет собой цинковый комплекс 13-(N-(2-(2-(2-N-(4-((4'-бром-2'-фторфенил)амино)-6-метоксихиназолин-7)-1-оксобутиратоаминоэтокси)этокси)этил))амид-17(3)-(1ʺ-дезокси-β-мальтозил-1ʺ-(1',2',3'-триазол-4'-ил)-метил)амидхлорина-e₆ общей формулы:at

is a zinc complex of 13- (N- (2- (2- (2-N- (4 - ((4'-bromo-2'-fluorophenyl) amino) -6-methoxyquinazolin-7) -1-oxobutyrato-amino-ethoxy) ethoxy ) ethyl)) amide-17 (3) - (1ʺ-deoxy-β-maltosyl-1ʺ (1 ', 2', 3'-triazol-4'-yl) -methyl) amidchlorin-e _{6 of the} general formula:

The task is achieved by the fact that the resulting derivative is used as an agent for photodynamic therapy.

FIG. 1 shows examples of synthesized purpurinimide-p6 conjugates with mono- and di-sugars.

FIG. 2 shows the synthesis schemes of Feoforbid-a (2), propargylamide feoforbid-a (3), zinc complex 13- (dimethylaminoethyl) amide-17 (3) -propargylamide chlorine-e ₆ (4).

FIG. 3 shows the synthesis schemes for 2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl azide (5), β-D-galactopyranosyl azide (6), 2,3,4,6-tetra-O- acetyl-β-D-glucopyranosyl azide (7), β-D-glucopyranosyl azide (8), hepta-O-acetyl-β-maltoazide (9), β-maltoazide (10).

FIG. 4 shows the synthesis of the zinc complex 13- (dimethylaminoethyl) amide-17 (3) - (tetra-O-acetyl-1ʺ-deoxy-β-D-galactopyranosyl-1ʺ- (1 ', 2', 3'-triazole-4 '-yl) -methyl) amidchlorin-e ₆ (11).

FIG. 5 shows the synthesis scheme for the zinc complex 13- (dimethylaminoethyl) amide-17 (3) - (tetra-O-acetyl-1ʺ-deoxy-β-D-glucopyranosyl-1ʺ- (1 ', 2', 3'-triazole-4 '-yl) -methyl) amidchlorin-e ₆ (12).

FIG. 6 shows the synthesis scheme of the zinc complex 13- (dimethylaminoethyl) amide-17 (3) - (hepta-O-acetyl-1ʺ-deoxy-β-maltosyl-1ʺ- (1 ', 2', 3'-triazole-4'- yl) -methyl) amidchlorin-e ₆ (13).

FIG. 7 shows the synthesis scheme for the zinc complex 13- (dimethyl aminoethyl) amide-17 (3) - (1ʺ-deoxy-β-D-galactopyranosyl-1ʺ- (1 ', 2', 3'-triazole-4'-yl) - methyl) amide chlorin-e ₆ (14), 13- (trimethylammonium ethyl) amide-17 (3) - (1ʺ-deoxy-β-D-galactopyranosyl-1ʺ- (1 ', 2', 3'-triazole- 4'-yl) -methyl) amidchlorin-e ₆ (15).

FIG. 8 shows the synthesis scheme of the zinc complex 13- (dimethylaminoethyl) amide-17 (3) - (1ʺ-deoxy-β-D-glucopyranosyl-1ʺ- (1 ', 2', 3'-triazol-4'-yl) -methyl ) amidchlorin-e ₆ (16), zinc complex bromide 13- (dimethylethylammonium ethyl) amide-17 (3) - (1ʺ-deoxy-β-D-glucopyranosyl-1ʺ- (1 ', 2', 3'-triazole- 4'-yl) -methyl) amidchlorin-e ₆ (17).

FIG. 9 shows the synthesis scheme for the zinc complex 13- (dimethylaminoethyl) amide-17 (3) - (1ʺ-deoxy-β-maltosyl-1ʺ- (1 ', 2', 3'-triazol-4'-yl) methyl) amidchlorin -e ₆ (18).

FIG. 10 shows the synthesis scheme for 13- (N- (2- (2- (2-aminoethoxy) ethoxy) ethyl)) amide-17 (3) -propargylamide chlorin-e ₆ (19), 13- (N- (2- (2- (2-N- (3-carboxy-1-oxopropyl) -aminoethoxy) ethoxy) ethyl)) amide-17 (3) -propargylamide chlorin-e ₆ (20).

FIG. 11 shows the synthesis scheme of the zinc complex 13- (N- (2- (2- (2-N- (4 - ((4'-bromo-2'-fluorophenyl) amino) -6-methoxy-quinazoline-7) -1-oxobutyratoethoxyethoxy) ) ethoxy) ethyl)) amide-17 (3) -propargylamide chlorin-e ₆ (22).

FIG. 12 shows the synthesis scheme for the zinc complex 13- (N- (2- (2- (2-N- (4 - ((4'-bromo-2'-fluorophenyl) amino) -6-methoxy-quinazoline-7) -1-oxobutyratoethoxyethoxy) ) ethoxy) ethyl)) amide-17 (3) - (1ʺ-deoxy-β-maltosyl-1ʺ- (1 ', 2', 3'-triazol-4'-yl) methyl) amidchlorin-e ₆ (23 ).

FIG. 13 shows the absorption spectra of compounds 15, 17, 18, 23 in water, the concentration of 5 μM.

FIG. 14 shows the fluorescence spectra of compounds 15, 17, 18, 23 in water, the concentration of 5 μM.

FIG. 15 shows the intracellular localization of compounds 15, 17, 18, 23 in cells of the A431 and CHO lines. Concentration 5 μm, incubation 4 hours. λEx. = 450 nm; λEm. = 600-740 nm.

FIG. 16 shows the fluorescence intensity of the cells after incubation with compounds 15, 17, 18, 23 at a concentration of 5 μM for 4 hours.

FIG. 17 shows the dependence of the viability of the A431 and CHO cell cultures on the concentration of compounds 15, 17, 18 in the dark and when irradiated at a dose of 20 J / cm ² .

FIG. 18 shows the dependence of the viability of A431 and CHO cell cultures on the concentration of compounds 23 in the dark and when irradiated at a dose of 20 J / cm ² .

The synthesis of the proposed derivative of the zinc metal complex of chlorin-e ₆ is as follows.

Synthesis Feoforbid-a (2) (Fig. 2). In a round bottom flask with a stirrer, 0.500 g of methylpheophorbide-1 was placed and 15 ml of acetone was added. Next, 0.7 ml of concentrated hydrochloric acid was added dropwise. The reaction mixture was allowed to stir at room temperature for 48 hours. The solvent was removed under reduced pressure. The residue was dissolved in 30 ml of chloroform, extracted with water (3 × 30 ml). The organic phase was dried over Na ₂ SO ₄ . After conducting column chromatography on silica gel (eluent: CHCl ₃ first, then 5% MeOH - 95% CHCl ₃ ), the pheophorbide-a (2) is obtained as a dark green powder (0.430 g, 89%). The compound is known [J. Org. Chem. 45 (1980) 1969-1674].

Synthesis of propargylamide-feophorbide-a (3) (Fig. 2). In a round-bottom flask "A" with a stirrer in an argon atmosphere, 0.430 g (1 equivalent) of pheophorbide-a 2, 0.284 g (2 equivalents) of N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (EDC⋅HCl) and 8 ml were placed CHCl ₃ . Stir at 0 ° C for 30 min. In a round-bottom flask “B” with a stirrer, 0.081 g (2 equivalents) of propargylamine, 0.045 g (0.5 equivalents) of DMAP and 8 ml of chloroform were mixed, the mixture was stirred for 10 minutes at 0 ° C. Then the syringe transferred the contents of flask “B” to flask “A” and stirred for 18 hours. The solvent was removed under reduced pressure. The residue was dissolved in 30 ml of chloroform, extracted with water (3 × 30 ml). The organic phase was dried over Na ₂ SO ₄ . After conducting column chromatography on silica gel (eluent: first CHCl ₃ , then 2% MeOH - 98% CHCl ₃ ), pheophorbide propargylamide (a) was isolated as a dark green powder (0.395 g, 87%).

Spectrum ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 9.49 (br.s, 2H), 8.63 (s, 1H), 7.98 (dd, J = 17.5, 11.5 Hz, 1H), 6.30 (d, J = 18.2 Hz, 1H), 6.27 (s, 1H), 6.20 (d, J = 19.9 Hz), 5.36 (br. s, 1H), 4.53 (br. s, 1H), 4.27 (br. s, 1H), 3.87 (s, 3H), 3.78-3.55 (m, 7H), 3.41 (s, 3H), 3.25 (s, 3H), 2.70 (br.s, 1H), 2.46 (br. s, 1H), 2.33-2.15 (m, 2H), 1.94 (s, 1H), 1.83 (d, J = 6.6 Hz, 3H), 1.68 (t, J = 7.6 Hz, 3H), -1.66 (br. S, 2H).

¹³ C NMR spectrum (101 MHz, CDCl ₃ , δ, ppm): 189.36, 172.27, 171.76, 169.56, 137.93, 128.86, 104.64, 104.26, 97.59, 79.16, 64.77, 52.97, 51.52, 50.25, 29.69, 28.83 , 23.31, 19.55, 17.19, 12.16, 11.26.

MS (MALDI): m / z 630.1 [M] +.

Synthesis of zinc complex 13- (dimethylaminoethyl) amide-17 (3) -propargylamide chlorin-e ₆ (4) (Fig. 2). In a round bottom flask with a stirrer, 0.395 g of compound 3 was placed and 16 ml of chloroform was added, then 2 ml of N, N-dimethylethylenediamine was added dropwise to the mixture. Stir at room temperature for 12 hours. Remove the solvent under reduced pressure. The residue was dissolved in 20 ml of chloroform, extracted with water (3 × 50 ml), the organic phase was dried over Na ₂ SO ₄ . The resulting organic phase was dissolved in 10 ml of chloroform and zinc diacetate (0.433 g, 5 equivalents) dissolved in 4 ml of methanol was added dropwise. The reaction mixture was stirred at room temperature for 1 hour. Then the solvent was removed under reduced pressure. The residue was dissolved in 30 ml of chloroform, extracted with water (3 × 30 ml). The organic phase was dried over Na ₂ SO ₄ . After carrying out column chromatography on silica gel (eluent: first 5% MeOH - 95% CHCl ₃ , then 15% MeOH - 85% CHCl ₃ ). The zinc complex of functionalized chlorin-e ₆ (4) was isolated as a dark green powder (0.402 g, 82%).

Spectrum ¹ H NMR (400 MHz, CDCl ₃ , δ, ppm): 9.51 (s, 2H), 8.96 (br.s, 1H), 8.66 (s, 1H), 8.34 (t, J = 5.4 Hz , 1H), 8.22 (dd, J = 17.8, 11.6 Hz, 1H), 6.22 (d, J = 17.9 Hz, 1H), 6.00 (d, J = 11.6 Hz, 1H), 5.41 (d, J = 19.1 Hz , 1H), 5.07 (d, J = 18.9 Hz, 1H), 4.42 (q, J = 7.0 Hz, 1H), 4.20 (d, J = 9.9 Hz, 1H), 3.91-3.75 (m, 6H), 3.69 (br.s, 4H), 3.37 (s, 3H), 3.35 (s, 3H), 3.26-3.16 (m, 3H), 3.05 (t, J = 2.3 Hz, 1H), 2.73 (br.s, 6H ), 2.47-2.38 (m, 1H), 2.07 (d, J = 8.8 Hz, 2H), 1.67 (t, J = 7.5 Hz, 3H), 1.61 (d, J = 7.0 Hz, 3H).

¹³ C NMR spectrum (101 MHz, CDCl ₃ , δ, ppm): 173.24, 171.72, 170.35, 165.14, 162.77, 151.66, 147.96, 146.14, 144.07, 143.10, 141.20, 140.74, 138.58, 137.23, 133.04, 132.29 , 130.66, 119.20, 101.79, 101.75, 99.87, 93.16, 81.19, 72.81, 52.24, 51.65, 46.26, 43.88, 37.46, 32.02, 29.99, 27.78, 22.80, 18.87, 17.89, 12.30, 11.76, 10.93.

MS (MALDI): m / z 779.0 [M] +.

Synthesis of 2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl azide (5) (Fig. 3). In a flask with a stirrer and filled with argon, 2 g of β-D-galactose pentaacetate was added and 20 ml of anhydrous dichloromethane was added, then a 1 M solution of tin tetrachloride in dichloromethane (2.56 ml, 0.5 equivalent) was added dropwise to the mixture then trimethylsilyl azide (0.82 ml, 1.2 equivalents). Stir at 0 ° C for 3 hours. The reaction mixture was then diluted with 80 ml of dichloromethane and washed with saturated NaHCO ₃ solution (70 ml), water (50 ml) and saturated sodium chloride solution (50 ml). The organic phase was dried over Na ₂ SO ₄ . After conducting column chromatography on silica gel (eluent: 50% PE - 50% EtOAc), 2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl azide (5) was isolated as a white crystalline powder (1.8 g, 95%).

Spectrum ¹ H NMR (300 MHz, CDCl ₃ , δ, ppm): 5.42 (dd, J = 3.3, 1.0 Hz, 1H), 5.16 (dd, J = 10.4, 8.7 Hz, 1H), 5.03 (dd , J = 10.3, 3.3 Hz, 1H), 4.59 (d, J = 8.7 Hz, 1H), 4.17 (dd, J = 6.5, 2.4 Hz, 2H), 4.05-3.98 (m, 1H), 2.17 (s, 3H), 2.09 (s, 3H), 2.06 (s, 3H), 1.99 (s, 3H).

¹³ C NMR spectrum (75 MHz, CDCl ₃ , δ, ppm): 170.51, 170.25, 170.13, 169.51, 88.44, 72.99, 70.85, 68.17, 66.97, 61.36, 20.82.

Synthesis of β-D-galactopyranosyl azide (6) (Fig. 3). 0.2 g of 2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl azide was placed in a round-bottom flask with a magnetic stirrer and dissolved in 4 ml of methanol. Then sodium methoxide was added dropwise until a strong alkaline medium was formed (pH = 11), stirred for 30 minutes. Then concentrated hydrochloric acid was added dropwise to the flask until neutral. The solvent was removed under reduced pressure. After conducting column chromatography on silica gel (eluent: first 20% CH3OH - 80% CHCl ₃ , then 50% CH ₃ OH - 50% CHCl ₃ ), β-D-galactopyranosyl azide (5) was isolated as a white crystalline powder (0944 g, 95%).

IR: 2100 cm-1 (band azide group).

Spectrum ¹ H NMR (400 MHz, CD ₃ OD, δ, ppm): 4.45 (d, J = 6.5 Hz, 1H), 3.89 (s, 1H), 3.84-3.71 (m, 2H), 3.65 ( d, J = 5.8 Hz, 1H), 3.56-3.48 (m, 2H).

¹³ C NMR spectrum (101 MHz, CD ₃ OD, δ, ppm): 92.61, 78.93, 74.97, 71.98, 70.23, 62.46.

Synthesis of 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl azide (7) (Fig. 3). In a Schlenk flask filled with argon with a stirrer, 2 g of glucose β-D-pentaacetate were added and 20 ml of anhydrous dichloromethane was added, then a 1 mol / l solution of tin tetrachloride in dichloromethane (2.56 ml, 0.5 equivalent) was added dropwise to the mixture, then trimethylsilyl azide (0.82 ml, 1.2 equivalents). Stir at 0 ° C for 3 hours. The reaction mixture was then diluted with 80 ml of dichloromethane and washed with saturated NaHCO ₃ solution (70 ml), water (50 ml) and saturated sodium chloride solution (50 ml). The organic phase was dried over Na ₂ SO ₄ . After silica gel column chromatography (eluent: 50% petroleum ether - 50% EtOAc), 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl azide (7) was isolated as a white non-crystalline powder (1 , 66 g, 87%).

Spectrum ¹ H NMR (300 MHz, CDCl ₃ , δ, ppm): 5.22 (t, J = 9.4 Hz, 1H), 5.10 (t, J = 9.7 Hz, 1H), 4.96 (t, J = 9.2 Hz, 1H), 4.65 (d, J = 8.8 Hz, 1H), 4.22 (ddd, J = 14.8, 12.5, 3.5 Hz, 2H), 3.79 (ddd, J = 9.9, 4.7, 2.3 Hz, 1H), 2.10 (s, 3H), 2.08 (s, 3H), 2.03 (s, 3H), 2.01 (s, 3H).

Spectrum ¹³ C NMR (75 MHz, CDCl ₃ , δ, ppm): 169.29, 169.19, 87.91, 74.01, 72.58, 70.61, 67.84, 61.64, 20.57.

Synthesis of β-D-glucopyranosyl azide (8) (Fig. 3). 0.3 g of 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl azide was placed in a round bottom flask with a magnetic stirrer and dissolved in 4 ml of methanol. Then sodium methoxide was added dropwise until a strong alkaline medium was formed (pH = 11), stirred for 30 minutes. Then concentrated hydrochloric acid was added dropwise to the flask until neutral. The solvent was removed under reduced pressure. After conducting column chromatography on silica gel (eluent: first 20% CH ₃ OH - 80% CHCl ₃ , then 50% CH ₃ OH - 50% CHCl ₃ ), β-D-glucopyranosyl azide (8) was isolated as a white non-crystalline powder (0.165 g, quantitatively).

IR: 2098 cm-1 (band azide group).

Spectrum ¹ H NMR (400 MHz, CD ₃ OD, δ, ppm): 4.50 (d, J = 8.5 Hz, 1H), 3.90 (d, J = 11.9 Hz, 1H), 3.70 (dd, J = 12.0, 5.3 Hz, 1H), 3.43-3.35 (m, 2H), 3.31 (d, J = 17.6 Hz, 2H), 3.16 (t, J = 8.7 Hz, 1H).

¹³ C NMR spectrum (101 MHz, CD ₃ OD, δ, ppm): 90.67, 78.76, 76.68, 73.35, 69.70, 61.12.

Synthesis of hepta-O-acetyl-β-maltoazide (9) (Fig. 3). In a Schlenk flask with a stirrer and filled with argon, 1.5 g of maltose β-D-octoacetate was added and 20 ml of anhydrous dichloromethane was added, then a 1 mol / l solution of tin tetrachloride in dichloromethane (1.1 ml, 0.5) was added dropwise to the mixture. equivalent), then trimethylsilyl azide (0.35 ml, 1.2 equivalents). Stir at 0 ° C for 3 hours. The reaction mixture was then diluted with 80 ml of dichloromethane and washed with saturated NaHCO ₃ solution (70 ml), water (50 ml) and saturated sodium chloride solution (50 ml). The organic phase was dried over Na ₂ SO ₄ . After column chromatography on silica gel (eluent 50% petroleum ether - 50% EtOAc), hepta-O-acetyl-β-maltoazide (9) was isolated as a white crystalline powder (1.66 g, 98%).

Spectrum ¹ H NMR (300 MHz, CDCl ₃ , δ, ppm): 5.39 (dd, J = 12.3, 7.0 Hz, 1H), 5.26 (t, J = 8.9 Hz, 1H), 5.05 (t, J = 9.9 Hz, 1H), 4.85 (dd, J = 10.5, 4.0 Hz, 1H), 4.82-4.68 (m, 2H), 4.50 (dd, J = 12.3, 2.5 Hz, 1H), 4.28-4.19 (m, 2H), 4.08-3.91 (m, 2H), 3.78 (ddd, J = 9.7, 4.3, 2.6 Hz, 1H), 2.15 (s, 3H), 2.10 (s, 3H), 2.06-1.98 (m, 15H) .

Spectrum ¹³ C NMR (75 MHz, CDCl ₃ , δ, ppm): 170.50, 170.39, 170.08, 169.92, 169.48, 169.39, 95.69, 87.46, 75.08, 74.23, 72.31, 71.47, 69.97, 69.24, 68.62, 67.92 , 62.51, 61.44, 20.85, 20.78, 20.69, 20.58.

Synthesis of β-maltozazide (10) (Fig. 3). In a round-bottom flask with a magnetic stirrer, 0.3 g of hepta-O-acetyl-β-maltosyl azide was placed and dissolved in 4 ml of methanol. Then sodium methoxide was added dropwise until a strong alkaline medium was formed (pH = 11), stirred for 30 minutes. Then concentrated hydrochloric acid was added dropwise to the flask until neutral. The solvent was removed under reduced pressure. After conducting column chromatography on silica gel (eluent: 50% CH ₃ OH - 50% CHCl ₃ ) (3-maltoazazide (10) was isolated as a white non-crystalline powder (0.165 g, 89%).

Spectrum ¹ H NMR (400 MHz, CD ₃ OD, δ, ppm): 5.19 (d, J = 3.8 Hz, 1H), 4.55 (d, J = 8.6 Hz, 1H), 3.93 (dd, J = 12.3, 1.6 Hz, 1H), 3.85 (dd, J = 12.6, 3.9 Hz, 2H), 3.74-3.55 (m, 5H), 3.53-3.43 (m, 2H), 3.37 (s, 1H), 3.28 (t , J = 9.3 Hz, 1H), 3.22 (t, J = 8.8 Hz, 1H).

¹³ C NMR spectrum (101 MHz, CD ₃ OD, δ, ppm): 102.90, 92.00, 80.65, 78.74, 77.87, 75.05, 74.82, 74.37, 74.14, 71.50, 62.73, 61.96.

Synthesis of the zinc complex of 13- (dimethylaminoethyl) amide-17 (3) - (tetra-O-acetyl-1--deoxy-β-D-galactopyranosyl-1ʺ (1 ', 2', 3'-triazole-4'-yl ) -methyl) amidhlorin-e6 (11) (Fig. 4). In a round bottom flask equipped with a reflux condenser, 0.06 g of compound 4 and 0.034 g (1.2 equivalents) of 2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl azide 5 were placed. The mixture was dissolved in 6 ml t-BuOH and 3 ml of CHCl ₃ . In a separate flask, 0.004 g of CuSO4⋅5H ₂ O, 0.006 g (0.4 equivalents) of sodium ascorbate (AscNa), 0.008 g (0.2 equivalents) of tris (1-benzyl-1H-1,2,3-triazole- 4-yl) methyl) amine (TBTA) and 6 ml of water; not dissolved solid particles were broken by ultrasound. After that, the prepared solution was added dropwise to the reaction mixture. The reaction mixture was stirred with a magnetic stirrer at 50 ° C for 1 hour. The solvent was removed under reduced pressure. The residue was dissolved in chloroform, extracted with water (3 × 30 ml). The organic phase was dried over Na ₂ SO ₄ . After column chromatography on silica gel (eluent: CHCl ₃ first, then 20% MeOH - 80% CHCl ₃ ), the target conjugate (11) was isolated as a dark green crystalline powder (0.063 g, 71%).

Spectrum ¹ H NMR (400 MHz, DMSO-d ₆ , δ, ppm): δ 9.52 (s, 1H), 9.50 (s, 1H), 8.80 (s, 1H), 8.65 (s, 1H), 8.39 (t, 1H), 8.22 (dd, J = 17.8, 11.6 Hz, 1H), 8.06 (s, 1H), 6.25-6.17 (m, 2H), 6.00 (d, 1H), 5.56 (t, 2H) , 5.51-5.34 (m, 2H), 5.09 (d, J = 18.5 Hz, 1H), 4.52 (t, 1H), 4.42 (dd, J = 14.3, 7.1 Hz, 1H), 4.31 (d, J = 5.6 Hz, 2H), 4.21 (d, J = 9.5 Hz, 1H), 4.08 (dd, J = 11.6, 5.1 Hz, 1H), 3.95 (dd, J = 11.5, 7.2 Hz, 1H), 3.80 (q, J = 15.4, 7.8 Hz, 3H), 3.65 (s, 3H), 2.86 (s, 2H), 2.17-2.04 (m, 6H), 1.92 (d, J = 1.9 Hz, 6H), 1.77 (s, 3H) , 1.66 (t, J = 14.2, 6.7 Hz, 3H), 1.60 (d, J = 7.8 Hz, 3H).

Spectrum ¹ H NMR (101 MHz, DMSO-d ₆ , δ, ppm): 173.58, 172.24, 170.20, 170.13, 169.69, 168.74, 165.31, 163.04, 151.80, 148.37, 146.26, 145.70, 144.20,143.37, 141.60 , 140.92, 138.79, 137.41, 133.27, 132.45, 130.92, 128.99, 127.97, 122.35, 119.41, 102.14, 100.12, 93.38, 84.38, 79.41, 73.11, 70.67, 69.18, 67.84, 67.53, 61.75, 52.60, 51.82, 45.18, 34.22 , 28.99, 23.07, 20.66, 20.60, 20.54, 20.22, 19.13, 18.14, 12.54, 11.94, 11.17.

MS (MALDI): m / z 1152.4 [M +].

Synthesis of 13- (dimethylaminoethyl) amide-17 (3) - (tetra-O-acetyl-1ʺ-deoxy-β-D-glucopyranosyl-1ʺ- (1 ', 2', 3'-triazol-4'-yl) zinc complex ) -methyl) amidchloro-on-e6 (12) (Fig. 5). In a round bottom flask equipped with a reflux condenser, 0.06 g of compound 4, 0.034 g (1.2 equivalents) of 2,3,4,6-tetra-O-acetyl-β-D-glucogranosyl azide 7 was placed. The mixture was dissolved in 6 ml t-BuOH and 3 ml of CHCl ₃ . In a separate flask, 0.004 g (2 equivalents) of CuSO ₄ ⋅5H ₂ O, 0.006 g (0.4 equivalents) of sodium ascorbate, 0.008 g (0.2 equivalents) of TBTA and 6 ml of water were mixed; not dissolved solid particles were broken by ultrasound. After that, the prepared solution was added dropwise to the reaction mixture. The reaction mixture was stirred with a magnetic stirrer at 50 ° C for 1 hour. The solvent was removed under reduced pressure. The residue was dissolved in chloroform, extracted with water (3 × 30 ml). The organic phase was dried over Na ₂ SO ₄ . After column chromatography on silica gel (eluent: CHCl ₃ first, then 20% MeOH - 80% CHCl _3. ), The target conjugate 12 was isolated as a dark green crystalline powder (0.06 g, 67%).

Spectrum ¹ H NMR (400 MHz, DMSO-d ₆ , δ, ppm): 9.51 (d, J = 5.6 Hz, 2H), 8.83 (s, 1H), 8.65 (s, 1H), 8.38 (t , J = 5.6 Hz, 1H), 8.22 (dd, J = 17.8, 11.5 Hz, 1H), 8.13 (s, 1H), 6.30-6.17 (m, 2H), 6.00 (dd, J = 11.5, 1.6 Hz, 1H), 5.64-5.37 (m, 4H), 5.18-5.02 (m, 2H), 4.42 (q, J = 7.1 Hz, 1H), 4.35-4.18 (m, 4H), 4.13-3.95 (m, 2H) , 3.81 (dd, J = 15.3, 7.6 Hz, 3H), 3.68 (s, 3H), 3.53 (d, J = 8.5 Hz, 1H), 2.92 (s, 1H), 2.00 (s, 6H), 1.92 ( d, J = 6.6 Hz, 6H), 1.74 (s, 3H), 1.67 (t, J = 8.5 Hz, 3H), 1.59 (d, J = 7.1 Hz, 3H).

Spectrum ¹ H NMR (101 MHz, DMSO-d ₆ , δ, ppm): 173.29, 171.90, 169.95, 169.52, 169.31, 168.41, 162.76, 151.93, 151.56, 145.62, 143.96, 143.11, 140.66, 138.54, 137.15 , 133.01, 132.21, 130.68, 121.78, 101.89, 83.70, 79.19, 73.17, 72.19, 70.00, 67.46, 61.78, 51.60, 33.98, 22.81, 20.42, 20.36, 20.22, 19.89, 18.88, 17.89, 12.30, 11.71, 10.93.

MS (MALDI): m / z 1152.1 [M +].

Synthesis of 13- (dimethylaminoethyl) amide-17 (3) - (hepta-O-acetyl-1ʺ-deoxy-β-maltosyl-1ʺ- (1 ', 2', 3'-triazol-4'-yl) -) zinc complex methyl) amidchlorin-e ₆ (13) (Fig. 6). In a round bottom flask equipped with a reflux condenser, 0.06 g of compound 4 and 0.061 g (1.2 equivalents) of hepta-O-acetyl-β-maltoazide 9 were placed. The mixture was dissolved in 6 ml of t-BuOH and 3 ml of CHCl ₃ . The mixture was dissolved in 6 ml of t-BuOH and 3 ml of CHCl ₃ . In a separate flask, 0.004 g (2 equivalents) of CuSO ₄ ⋅5H ₂ O, 0.006 g (0.4 equivalents) of sodium ascorbate, 0.008 g (0.2 equivalents) of TBTA and 6 ml of water were mixed; not dissolved solid particles were broken by ultrasound. After that, the prepared solution was added dropwise to the reaction mixture. The reaction mixture was stirred with a magnetic stirrer at 50 ° C for 1 hour. The solvent was removed under reduced pressure. The residue was dissolved in chloroform, extracted with water (3 × 30 ml). The organic phase was dried over Na ₂ SO ₄ . After column chromatography on silica gel (eluent: CHCl ₃ first, then 20% MeOH - 80% CHCl ₃ ), the target conjugate (13) was isolated as a dark green crystalline powder (0.07 g, 64%).

Spectrum ¹ H NMR (400 MHz, DMSO-d ₆ , δ, ppm): 9.51 (d, J = 6.1 Hz, 2H), 8.80 (s, 1H), 8.65 (s, 1H), 8.37 (t , J = 5.3 Hz, 1H), 8.33 (s, 1H), 8.22 (dd, J = 17.7, 11.6 Hz, 1H), 8.04 (s, 1H), 6.28-6.16 (m, 2H), 6.00 (d, J = 11.5 Hz, 1H), 5.62-5.41 (m, 3H), 5.37-5.18 (m, 2H), 5.14-4.85 (m, 3H), 4.47-3.94 (m, 10H), 3.81 (d, J = 7.5 Hz, 2H), 3.68 (s, 3H), 3.53 (d, J = 7.9 Hz, 2H), 2.86 (s, 2H), 2.03-1.87 (m, 21H), 1.76-1.52 (m, 12H).

Spectrum ¹ H NMR (101 MHz, DMSO-d ₆ , δ, ppm): 173.30, 171.92, 170.05, 169.96, 169.82, 169.66, 169.50, 169.15, 168.63, 165.05, 162.75, 151.55, 148.11, 146.01, 145.54 , 143.95, 143.12, 141.34, 140.65, 138.54, 137.15, 136.22, 133.80, 133.01, 132.19, 130.68, 128.73, 127.72, 121.83, 119.12, 101.90, 101.62, 99.88, 95.72, 93.11, 83.29, 79.19, 74.43, 73.77, 73.33 , 70.54, 69.39, 68.90, 68.13, 67.67, 62.82, 61.35, 57.64, 52.33, 51.58, 46.35, 44.79, 37.40, 33.97, 32.13, 30.08, 22.82, 20.49, 20.42, 20.35, 20.31, 20.27, 19.90, 18.88, 17.89 , 12.30, 11.70, 10.93.

MS (MALDI): m / z 1442.3 [M +].

Synthesis of the zinc complex of 13- (dimethylaminoethyl) amide-17 (3) - (1--deoxy-β-D-galactopyranosyl-1 ,- (1 ', 2', 3'-triazol-4'-yl) -methyl) amide chlorin- e ₆ (14) (Fig. 7). 0.1 g of compound 4 and 0.031 g (1.2 equivalents) of β-D-galactopyranosyl azide 6 were placed in a round bottom flask equipped with a reflux condenser, the mixture was dissolved in 5 ml of dimethylformamide. In another round-bottomed flask were mixed 0.010 g (2 equivalents) CuSO ₄ ⋅5H ₂ O, 0,010 g (0.4 equivalents) of sodium ascorbate, 0.014 g (0.2 equivalents) TVTA and 5 ml of water, ultrasound was used to accelerate the dissolution. After that, the prepared solution was added dropwise to the reaction mixture. The reaction mixture was stirred with a magnetic stirrer at 50 ° C for 1 hour. The solvent was removed under reduced pressure. After conducting column chromatography on silica gel (eluent: first 20% CH ₃ OH - 80% CHCl ₃ , then 50% CH ₃ OH - 50% CHCl ₃ ), the target conjugate (14) was isolated as a dark green crystalline powder (0.104 g, 83%).

¹ H NMR spectrum (400 MHz, CD ₃ OD, δ, ppm): 9.56 (d, J = 12.7 Hz, 2H), 8.64 (s, 1H), 8.12 (dd, J = 17.8, 11.5 Hz, 1H), 7.90 (d, J = 6.1 Hz, 1H), 6.18 (d, J = 18.3 Hz, 1H), 6.00 (d, J = 11.4 Hz, 1H), 5.51-5.36 (m, 2H), 5.19 ( d, J = 18.8 Hz, 1H), 4.43 (d, J = 23.5, 11.6 Hz, 1H), 4.31 (q, J = 15.4 Hz, 3H), 4.01 (t, J = 9.4 Hz, 1H), 3.91- 3.79 (m, 3H), 3.75 (s, 3H), 3.73 (d, J = 7.6 Hz, 1H), 3.70-3.65 (m, 2H), 3.65-3.56 (m, 6H), 3.40 (s, 3H) , 3.37 (s, 3H), 3.34 (s, 3H), 3.33 (s, J = 5.0 Hz, 3H), 2.63 (s, 2H), 2.49-2.39 (m, 1H), 2.25 (s, 6H), 2.11-2.00 (m, 1H), 1.98-1.83 (m, 1H), 1.73 (s, 3H), 1.66 (d, J = 7.1 Hz, 3H).

Spectrum ¹ H NMR (101 MHz, CD ₃ OD, δ, ppm): 175.06, 174.12, 165.70, 161.99, 152.80, 147.34, 145.21, 144.92, 143.60, 141.34, 141.24, 138.88, 138.12, 132.98, 132.64, 130.43, 121.43, 119.14, 118.42, 102.25, 101.36, 99.88, 92.71, 88.70, 78.36, 73.80, 69.87, 68.83, 60.83, 52.59, 51.35, 44.10, 37.83, 37.48, 34.13, 32.11, 30.21, 29.24, 22.04, 18.84, 16.75, 10.96, 10.58, 9.71.

MS (MALDI): m / z 983.6. [M +].

Synthesis of zinc complex bromide 13- (dimethylethylammonium) amide-17 (3) - (1--deoxy-β-D-galactopyranosyl-1ʺ- (1 ', 2', 3'-triazol-4'-yl) methyl) amide chloride -e ₆ , (15) (Fig. 7). In a round bottom flask with a magnetic stirrer, 0.050 g of compound 14 was placed, dissolved in a mixture of 2 ml of chloroform and 3 ml of methanol, and 2 ml of ethyl bromide was added. The reaction mixture was stirred at room temperature for 48 hours. The solvent was removed under reduced pressure. 0.051 g (quantitatively) of the ammonium salt 15 of a dark green solid crystalline powder was isolated.

Spectrum ¹ H NMR (400 MHz, CD ₃ OD, δ, ppm): 9.60 (d, J = 24.9 Hz, 2H), 8.69 (s, J = 8.0 Hz, 1H), 8.21-8.06 (m, 1H), 7.97 (s, 1H), 6.20 (d, J = 17.6 Hz, 1H), 6.03 (d, J = 10.3 Hz, 1H), 5.46 (d, J = 9.2 Hz, 1H), 5.42-5.07 ( m, 2H), 4.53-4.44 (m, 1H), 4.39-4.27 (m, 3H), 4.07 (t, J = 9.3 Hz, 1H), 3.95-3.78 (m, 5H), 3.73 (d, J = 4.3 Hz, 5H), 3.64 (d, J = 4.0 Hz, 4H), 3.10-2.87 (m, 6H), 2.60-2.48 (m, 1H), 2.32-2.21 (m, 1H), 2.21-2.10 (m , 1H), 1.97-1.86 (m, 1H), 1.77 (q, J = 7.3 Hz, 3H), 1.71 (d, J = 7.0 Hz, 3H).

Spectrum ¹ H NMR (101 MHz, CD ₃ OD, δ, ppm): δ 175.95, 175.52, 172.99, 167.43, 163.64, 154.51, 149.15, 146.95, 146.37, 145.06, 143.00, 140.31, 139.75, 134.61, 134.39 , 131.98, 131.74, 122.89, 120.09, 103.98, 102.59, 101.34, 94.33, 90.12, 79.81, 75.21, 71.33, 70.29, 62.31, 52.88, 51.01, 44.49, 39.81, 35.61, 33.52, 31.49, 23.44, 20.86, 20.28, 18.20 , 14.46, 12.38, 12.19, 11.14, 8.38.

Synthesis of the zinc complex of 13- (dimethylaminoethyl) amide-17 (3) - (1ʺ-deoxy-β-D-glucopyranosyl-1ʺ- (1 ', 2', 3'-triazol-4'-yl) methyl) amide chlorin- e ₆ (16) (Fig. 8). In a round bottom flask equipped with a reflux condenser, 0.123 g of compound 4 and 0.039 g (1.2 equivalents) of β-D-glucopyranosyl azide 8 were placed, the mixture was dissolved in 5 ml of dimethylformamide. In another round-bottomed flask, 0.008 g (2 equivalents) of CuSO ₄ ⋅ 5H ₂ O, 0.012 g (0.4 equivalents) of sodium ascorbate, 0.016 g (0.2 equivalents) of TBTA and 5 ml of water were mixed, and ultrasound was used to accelerate dissolution. After that, the prepared solution was added dropwise to the reaction mixture. The reaction mixture was stirred with a magnetic stirrer at 50 ° C for 1 hour. The solvent was removed under reduced pressure. After conducting column chromatography on silica gel (eluent: first 20% CH ₃ OH - 80% CHCl ₃ , then 50% CH ₃ OH - 50% CHCl ₃ ), the target conjugate (16) was isolated as a dark green crystalline powder (0.106 g, 69%).

¹ H NMR spectrum (400 MHz, CD ₃ OD, δ, ppm): δ 9.58 (d, J = 13.4 Hz, 2H), 8.66 (s, 1H), 8.14 (dd, J = 17.9, 11.5 Hz , 1H), 7.86 (s, 1H), 6.20 (d, J = 17.9 Hz, 1H), 6.02 (d, J = 10.2 Hz, 1H), 5.52-5.41 (m, 2H), 5.22 (d, J = 19.0 Hz, 1H), 4.46 (d, J = 7.2 Hz, 1H), 4.32 (d, J = 2.7 Hz, 3H), 3.90-3.74 (m, 8H), 3.52-3.34 (m, 12H), 2.72 ( s, 2H), 2.52-2.39 (m, 1H), 2.33 (s, 6H), 2.31-2.20 (m, 1H), 2.13-2.03 (m, 1H), 1.98-1.87 (m, 1H), 1.75 ( t, J = 7.5 Hz, 3H), 1.69 (d, J = 7.2 Hz, 3H).

¹³ C NMR spectrum (101 MHz, CD ₃ OD, δ, ppm): 176.70, 175.75, 174.42, 167.35, 163.65, 154.44, 149.33, 148.99, 146.86, 146.37, 145.25, 142.98, 142.91, 140.54, 139.77, 134.65, 134.30, 133.43, 132.07, 123.55, 120.07, 103.90, 103.01, 101.53, 94.38, 89.69, 81.23, 78.62, 74.11, 71.03, 62.51, 59.21, 54.21, 53.00, 45.70, 39.48, 39.05, 35.72, 33.73, 31.82, 23.70, 20.49, 18.40, 12.61, 12.23, 11.36.

MS (MALDI): m / z 984.3. [M +].

Synthesis of zinc complex bromide 13- (dimethylethylammonium) amide-17 (3) - (1--deoxy-β-D-glucopyranosyl-1ʺ (1 ', 2', 3'-triazol-4'-yl) -methyl) amide chloride -on-e6 (17) (Fig. 8). In a round bottom flask with a magnetic stirrer, 0.050 g of compound 16 was placed, dissolved in a mixture of 2 ml of chloroform and 3 ml of methanol, and 2 ml of ethyl bromide was added. The reaction mixture was stirred at room temperature for 48 hours. The solvent was removed under reduced pressure. 0.051 g (quantitatively) of ammonium salt 17 was isolated as a dark green solid crystalline powder.

¹ H NMR spectrum (400 MHz, CD ₃ OD, δ, ppm): 9.59 (d, J = 23.1 Hz, 2H), 8.67 (s, J = 5.3 Hz, 1H), 8.13 (dd, J = 17.9, 11.6 Hz, 1H), 7.91 (s, 1H), 6.20 (d, J = 17.3 Hz, 1H), 6.03 (d, J = 11.5 Hz, 1H), 5.50 (d, J = 9.2 Hz, 1H) , 5.42-5.09 (m, 2H), 4.48 (d, J = 7.1 Hz, 1H), 4.34 (s, 3H), 3.90-3.56 (m, 13H), 3.16 (s, 4H), 3.04 (s, J = 15.4 Hz, 2H), 2.59-2.47 (m, 1H), 2.33-2.21 (m, 1H), 2.16-2.07 (m, 1H), 1.97-1.85 (m, 1H), 1.76 (t, J = 7.5 Hz, 3H), 1.70 (d, J = 7.1 Hz, 3H).

¹³ C NMR spectrum (101 MHz, CD ₃ OD, δ, ppm): 175.89, 175.43, 170.66, 167.30, 163.50, 154.40, 146.10, 142.87, 140.21, 134.26, 131.64, 123.23, 119.96,108.20 103.33, 102.48, 101.20, 93.86, 89.39, 80.94, 78.31, 73.83, 70.76, 62.21, 53.78, 52.78, 51.01, 44.17, 42.92, 39.95, 39.70, 37.06, 35.46, 33.39, 31.35, 23.36, 20.17, 18.09, 12.38, 12.08, 11.04, 8.35.

Synthesis of Zinc Complex 13- (dimethylaminoethyl) amide-17 (3) - (1ʺ-deoxy-β-maltosyl-1 (- (1 ', 2', 3'-triazole-4'-yl) methyl) amidchlorin-e ₆ (18) (Fig. 9). In a round bottom flask equipped with a reflux condenser, 0.1 g of compound 4 and 0.055 g (1.2 equivalents) of β-D-glucopyranosyl azide 10 were placed. The mixture was dissolved in 5 ml of DMF. While the mixture was stirred, CuSO ₄ ⋅5H ₂ O 0.007 g (2 eq.), 0.010 g (0.4 equivalents) of sodium ascorbate, 0.014 g (0.2 equivalents) of TVTA and 5 ml of water were mixed in another round-bottomed flask. dissolution acceleration used ultrasound. After that, the prepared catalyst was added dropwise to the reaction mixture. The reaction mixture was stirred with a magnetic stirrer at 50 ° C for 1 hour. The solvent was removed under reduced pressure. After conducting column chromatography on silica gel (eluent: first, 1% triethylamine - 10% MeOH - 89% CHCl ₃ , then 3% triethylamine -20% MeOH - 77% CHCl _3. ) The target conjugate (18) is isolated as a dark green crystalline powder (0.074 g, 50%).

¹ H NMR spectrum (400 MHz, CD ₃ OD, δ, ppm): 9.58 (d, J = 19.1 Hz, 2H), 8.66 (s, 1H), 8.14 (dd, J = 17.8.11.6 Hz, 1H), 7.89 (s, 1H), 6.21 (dd, J = 17.9, 1.6 Hz, 1H), 6.04 (dd, J = 11.5, 1.6 Hz, 1H), 5.56-5.40 (m, 2H), 5.32-5.15 (m, 2H), 4.48 (q, J = 7.0 Hz, 1H), 4.33 (s, 3H), 4.09-3.99 (m, 1H), 3.97-3.80 (m, 6H), 3.75 (s, 7H), 3.72-3.54 (m, 8H), 3.46 (s, 3H), 3.40-3.38 (m, 3H), 3.36 (s, J = 5.8 Hz, 3H), 2.96 (s, 6H), 2.57-2.43 (m, 1H), 2.37-2.22 (m, 1H), 2.17-2.05 (m, 1H), 2.01-1.86 (m, 1H), 1.76 (t, J = 7.5 Hz, 3H), 1.70 (d, J = 7.1 Hz 3H).

Spectrum ¹³ C NMR (101 MHz, CD ₃ OD, δ, ppm): 176.70, 175.75, 167.50, 154.60, 149.20, 147.04, 146.39, 145.26, 143.04, 142.81, 140.65, 139.87,134.71, 134.44, 132.02, 123.51, 120.18, 104.06, 103.12, 102.94, 101.54, 94.42, 89.47, 80.48, 79.69, 78.34, 75.26, 75.06, 74.35, 73.68, 71.69, 62.93, 61.92, 59.21, 53.06, 45.17, 39.82, 38.34, 35.72, 33.66, 32.97, 31.76, 23.92, 23.71, 20.49, 18.40, 12.61, 12.31, 11.36, 9.43.

MS (MALDI): m / z 1145.8. [M +].

Synthesis of the 13- (N- (2- (2- (2-aminoethoxy) ethoxy) ethyl)) amide-17 (3) -propargylamide chlorin-e ₆ (19) zinc complex (Fig. 10). In a round bottom flask with a stirrer, 0.208 g of compound 3 was added and 8 ml of chloroform was added, then 1 ml of 1,2-bis (2-aminoethoxy) ethane was added dropwise to the mixture and stirred at room temperature for 12 hours. The solvent was removed under reduced pressure . The residue was dissolved in 20 ml of chloroform, extracted with water (3 × 50 ml), the organic phase was dried over Na ₂ SO ₄ . The resulting organic phase was dissolved in 7 ml of chloroform and a solution of 0.325 g of zinc diacetate in 4 ml of methanol was added dropwise. The reaction mixture was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure. The residue was dissolved in 30 ml of chloroform, extracted with water (3 × 30 ml). The organic phase was dried over Na ₂ SO ₄ . The target aminochlorin (19) is isolated in the form of a dark green powder (0.200 g, 72%). Used further without further purification.

Synthesis of the 13- (N- (2- (2- (2-N- (3-carboxy-1-oxopropyl) -aminoethoxy) ethoxy) ethyl)) amide-17 (3) -propargylamidyl-chlorin-e6 (20) ( Fig. 10). In a round bottom flask with a stirrer, 0.170 g of compound 19 was placed and 8 ml of chloroform was added, then 0.026 g (1.3 equivalents) of succinic anhydride and 0.026 g (1 equivalent) of diisopropylethylamine were added to the mixture. Stir at room temperature for 3 h. Solvent was removed under reduced pressure. The residue was dissolved in 20 ml of chloroform, extracted with water (3 × 50 ml), the organic phase was dried over Na ₂ SO ₄ . After conducting column chromatography on silica gel (eluent: 30% MeOH - 70% CHCl ₃ ), the target functionalized chlorine (20) was isolated as a dark green crystalline powder (0.190 g, quantitatively).

Spectrum ¹ H NMR (400 MHz, DMSO-d ₆ , δ, ppm): 9.50 (d, J = 6.3 Hz, 2H), 8.84 (s, 1H), 8.33 (t, J = 5.4 Hz, 1H ), 8.22 (dd, J = 17.8, 11.6 Hz, 1H), 7.95 (t, J = 5.3 Hz, 1H), 6.21 (d, J = 17.9 Hz, 1H), 6.00 (d, J = 11.6 Hz, 1H ), 5.43 (d, J = 18.7 Hz, 1H), 5.07 (d, J = 19.0 Hz, 1H), 4.42 (q, J = 7.2 Hz, 1H), 4.20 (d, J = 9.9 Hz, 1H), 3.88-3.77 (m, 6H), 3.72 (s, J = 4.8 Hz, 3H), 3.69-3.60 (m, 7H), 3.48 (t, J = 5.9 Hz, 2H), 3.04 (s, 1H), 2.39 (d, J = 6.2 Hz, 3H), 2.33 (d, J = 6.1 Hz, 3H), 2.07 (d, J = 8.6 Hz, 2H), 1.67 (t, J = 7.5 Hz, 3H), 1.60 (d J = 7.0 Hz, 3H).

¹³ C NMR spectrum (400 MHz, DMSO-d ₆ , δ, ppm): 206.52, 173.30, 171.74, 171.15, 170.10, 165.01, 162.77, 151.53, 148.17, 145.98, 143.92, 143.13, 141.40, 140.64, 138.64 , 137.15, 133.93, 133.00, 132.17, 130.70, 119.13, 101.94, 101.63, 99.90, 93.12, 81.20, 72.81, 69.73, 69.66, 69.22, 68.95, 52.29, 51.60, 46.26, 37.32, 32.07, 30.71, 30.05, 29.31, 27.79 , 22.84, 18.89, 17.91, 12.32, 11.65, 10.94.

MS (MALDI): m / z 939.8. [M +].

Synthesis of the zinc complex 13- (N- (2- (2- (2-N- (4 - ((4'-bromo-2'-fluorophenyl) amino) -6-methoxy-quinazolin-7) -1-oxobutyrato-amino-ethoxy) ethoxy) -ethyl)) amide-17 (3) -propargylamide chlorin-e ₆ (22) (Fig. 11). 0.095 g of compound 20, 0.038 g (2 equivalents) of EDC⋅HCl and 4 ml of dimethylformamide were placed in the first round-bottom flask with a stirrer. Stir at 0 ° C for 30 min. 0.073 g (2 equivalents) of 4 - ((4'-bromo-2'-fluorophenyl) amino) -6-methoxyquinazolin-7-ol (21), which was previously synthesized according to a procedure known in the art, was added to the second flask with a stirrer. from WO 2010/028254 A2, 0.006 g (0.5 equivalent) of DMAP and 3 ml of chloroform, the mixture was stirred for 10 minutes at 0 ° C. Then the syringe transferred the contents of the second flask to the first and stirred for 18 hours. The solvent was removed under reduced pressure. After column chromatography on silica gel (eluent: CHCl ₃ first, then 4% MeOH - 96% CHCl ₃ ), the target conjugate (22) was isolated as a dark green powder (0.075 g, 58%).

1 H NMR (400 MHz, DMSO-d ₆ , δ, ppm): δ 9.75 (s, 1H), 9.51 (d, J = 5.0 Hz, 2H), 8.84 (s, 1H), 8.65 (s , 1H), 8.32 (t, J = 5.4 Hz, 1H), 8.27-8.17 (m, 2H), 8.09 (t, J = 5.5 Hz, 1H), 7.96 (s, 1H), 7.71-7.65 (m, 1H) 7.54-7.43 (m, 2H), 7.36 (s, 1H), 6.21 (d, J = 19.1 Hz, 1H), 6.00 (d, J = 12.7 Hz, 1H), 5.43 (d, J = 19.1 Hz, 1H), 5.07 (d, J = 18.7 Hz, 1H), 4.41 (q, J = 7.2 Hz, 1H), 4.20 (d, J = 9.6 Hz, 1H), 3.92 (s, 3H), 3.88- 3.77 (m, 5H), 3.75-3.70 (m, 2H), 3.67 (s, 5H), 3.52 (t, J = 5.8 Hz, 2H), 3.37 (s, 3H), 3.05 (t, J = 2.4 Hz , 1H), 2.86 (t, J = 6.9 Hz, 2H), 2.55 (t, J = 7.0 Hz, 2H), 2.06 (d, J = 8.9 Hz, 2H), 1.66 (t, J = 7.5 Hz, 3H ), 1.60 (d, J = 7.0 Hz, 3H).

¹³ C NMR spectrum (400 MHz, DMSO-d ₆ , δ, ppm): δ 173.28, 171.70, 170.49, 170.46, 170.07, 165.00, 162.77, 157.95, 157.31, 155.45, 153.16, 151.52, 150.23, 148.17, 145.97, 145.18, 145.13, 143.91, 143.12, 141.40, 140.64, 138.67, 137.14, 133.93, 132.99, 132.15, 130.69,129.65, 127.58, 126.01, 125.89, 120.63, 119.53, 119.29, 119.12, 118.10, 118.01, 119.10, 119.47, 101.94, 101.62, 99.90, 93.11, 81.18, 79.17, 72.80, 69.74, 69.65, 69.22, 68.94, 56.56, 52.28, 51.57, 46.23, 37.29, 32.07, 30.03, 29.82, 28.94, 27.78, 22.82, 18.86, 17.88, 12.31, 11.63, 10.92.

MS (MALDI): m / z 1286.8. [M +].

Synthesis of the zinc complex 13- (N- (2- (2- (2-N- (4 - ((4'-bromo-2'-fluorophenyl) amino) -6-methoxy-quinazolin-7) -1-oxobutyrato-amino-ethoxy) ethoxy) ethyl)) amide-17 (3) - (1ʺ-deoxy-β-maltosyl-1ʺ- (1 ', 2', 3'-triazol-4'-yl) -methyl) amidchlorin-e ₆ (23) ( . 12). In a round bottom flask equipped with a reflux condenser, 0.058 g of compound 22, 0.02 g (1.2 equivalents) of β-maltoazide 10 were placed, the mixture was dissolved in 6 ml of dimethylformamide. In another round-bottomed flask, 0.002 g (0.2 equivalents) of CuSO ₄ ⋅ 5H ₂ O, 0.003 g (0.4 equivalents) of sodium ascorbate, 0.005 g (0.2 equivalents) of TBTA, and 6 ml of water were mixed, and ultrasound was used to accelerate dissolution . After that, the prepared catalyst was added dropwise to the reaction mixture. The reaction mixture was stirred at 50 ° C for 1 hour. The solvent was removed under reduced pressure. After conducting column chromatography on silica gel (eluent: first 20% MeOH - 80% CHCl ₃ , then 40% MeOH - 60% CHCl ₃ ), the target conjugate (23) was isolated as a dark green crystalline powder (0.066 g, 88%).

Spectrum ¹ H NMR (400 MHz, DMSO-d ₆ , δ, ppm): δ 9.93 (s, 1H), 9.50 (d, J = 4.8 Hz, 2H), 8.84 (s, 1H), 8.65 ( s, 1H), 8.38-8.31 (m, 1H), 8.26-8.17 (m, 2H), 8.11 (t, J = 5.6 Hz, 1H), 8.05 (s, 1H), 8.00 (s, 1H), 7.66 (dd, J = 9.9, 1.9 Hz, 1H), 7.52-7.43 (m, 2H), 7.36 (s, 1H), 6.21 (d, J = 19.4 Hz, 1H), 5.99 (d, J = 11.5 Hz, 1H), 5.72 (d, J = 2.9 Hz, 1H), 5.66-5.38 (m, 5H), 5.16-4.88 (m, 5H), 4.59-4.49 (m, 3H), 4.42 (q, J = 6.7 Hz , 1H), 4.36-4.18 (m, 3H), 3.93 (s, 3H), 3.86-3.41 (m, 30H), 3.13-3.03 (m, 2H), 2.86 (t, J = 6.9 Hz, 2H), 2.55 (t, J = 7.0 Hz, 2H), 1.66 (t, J = 7.5 Hz, 3H), 1.59 (d, J = 7.0 Hz, 3H).

¹³ C NMR spectrum (400 MHz, DMSO-d ₆ , δ, ppm): δ 173.33, 171.92, 170.53, 170.47, 170.11, 165.08, 162.75,157.35, 153.18, 151.54, 150.21, 148.16, 145.98, 145.18, 145.12, 144.84, 143.91, 143.12, 141.41, 140.64, 138.66, 137.14, 133.91, 132.98, 132.17, 130.70, 129.67, 127.55, 121.86, 120.58, 119.50, 113.15, 103.77, 101.95, 100.91, 87.06, 79.11, 77.92, 76.65, 76.25, 73.53, 73.22, 72.41, 71.52, 69.85, 69.74, 69.66, 69.22, 68.96, 60.74, 60.23, 56.67, 51.62, 34.07, 32.12, 29.84, 28.96, 22.86,18.88, 17.89, 12.32, 11.64, 10.93.

MS (MALDI): m / z 1656.1. [M +].

Analysis of the spectral properties of the compounds obtained.

The absorption and fluorescence spectra were recorded using a Synergy MX plate spectrophotometer-spectrofluorimeter (BioTek, USA). For the analysis, solutions of chlorin-e ₆ derivatives at a concentration of 5 μM were used. Deionized water was used as a solvent. Absorption spectra were recorded in the wavelength range of 300–700 nm with a step of 2 nm. Fluorescence spectra were recorded at 410 nm excitation, 2 nm pitch, recording range 550-850 nm.

The molar extinction coefficient (ε) was determined by the formula:

ε = D / cl, where:

D is the optical density, c is the concentration of the substance, l is the path length.

The fluorescence quantum yield of the test compounds was calculated relative to rhodamine B (Sigma, USA) in water (0.31). Calculations were made according to the formula:

Where:

ϕ1 - quantum yield of fluorescence of the test compound,

ϕ2 is the fluorescence quantum yield of the standard compound;

D1 and D2 are the optical densities of the reference and test solutions, respectively.

It is shown that the compounds 15, 17, 18, 23 under study are characterized by intense absorption in the shortwave region (Soret band) and in the longwave region (Q-band) (Fig. 13). The absorption maxima were 410 nm and 635 nm, respectively. The molar extinction coefficient in the short-wave part of the spectrum is much higher, which is typical of chlorin-type chromophores (Ferreira et al., 2008; Ormond, Freeman, 2013). For compound 23 (with digalactosyl and vandetanib), the peaks are shifted to the long-wavelength part of the spectrum (420 nm and 645 nm).

The fluorescence of chlorin-e ₆ derivatives is recorded in the long-wavelength region of the 620-700 nm spectrum (Fig. 14), which is characteristic of chlorinated compounds. The peak fluorescence intensity is at 645 nm.

Based on the recorded fluorescence spectra and the extinction values of the solutions, the values of relative fluorescence quantum yield were calculated, presented in Table 1. Rhodamine B. was used as a compound with a known fluorescence quantum yield.

For compound 23, the shift of the peaks in the absorption spectrum, the lowest fluorescence intensity, and a low level of quantum fluorescence are shown. This may be due to the aggregation of the compound molecules in an aqueous solution.

Cell culture.

Studies of the intracellular localization and photodynamic activity of the compounds were carried out on cell cultures:

Line A-431 - epidermal carcinoma of human skin (ATCC® CRL-1555 ™), with a high level of expression of the epidermal growth factor receptor (EGFR);

The CHO line is a Chinese hamster ovary cell (ATCC® CRL-9618TM). Cells of non-neoplastic nature, expression of EGFR is absent.

Cells were grown on nutrient medium MEMA NEEDLE with 10% fetal calf serum (HyClone, USA) and 2 mM L-glutamine (PanEco, Russia) at 37 ° C and 5% CO2 atmosphere. Cultivation was carried out in culture flasks with an area of 25 cm ² (Corning, USA). Cells were transplanted 3 times a week at a ratio of 1: 7. The cells were removed from the culture flask with trypsin: EDTA (1: 1) solution (PanEco, Russia); 10 mM phosphate-saline buffer (PBS) was used to wash the cells. Cell centrifugation was performed on a 541 OR centrifuge (Eppendorf, Germany). The condition of the cells was assessed using an Axiovert 200 inverted microscope (Carl Zeiss, Germany).

Analysis of the intracellular localization of compounds.

To study the intracellular localization of the test compounds, a cell suspension was planted in 96-well thin-walled plates (Corning, USA) at a concentration of 9 × 103 cells per well and incubated for 24 hours at 37 ° C and 5% CO ₂ atmosphere. Then the nutrient medium was replaced with a solution of the test compound in serum-free medium at a concentration of 5 μM, 200 μl / well, and incubated for 4 hours. Upon completion of the incubation, the cells were washed three times from the photosensitizer and fixed with 4% formaldehyde solution for 30 minutes.

Images of cells were obtained using an Axio Observer Z1 LSM 710 laser scanning confocal microscopy system (Carl Zeiss, Germany). Fluorescence was excited at a wavelength of 405 nm and recorded in the range of 600-740 nm.

On the basis of the images obtained, an analysis of the entry of compounds into the cells was performed. Quantitative assessment of the fluorescence intensity of the cells was carried out using the program ZEN 2012 (Carl Zeiss, Germany).

The study of the intracellular localization of compounds was carried out on cells with different levels of expression of the epidermal growth factor receptor (EGFR). It has been suggested that compound 23 (with digalactosyl and vandetanib) can selectively accumulate in cells with a high level of EGFR expression. For this purpose, a CHO line was selected for the study, the cells of which do not express either EGFR or VEGFR (Colagar et al., 2013) and the A431 line, whose cells are characterized by EGFR overexpression (Stanton et al., 1994). The expression level of EGFR in cell cultures used in the laboratory was previously verified by flow cytometry with staining antibodies specific for this receptor (Otvagin et al., 2018).

Analysis of the fluorescence of the cells after incubation with the test compounds showed that compounds 15, 17, 18, 23 quickly accumulate in the cells (Fig. 15). The nature of the distribution suggests that all compounds are predominantly localized in the cytoplasm in the intracellular membranes.

The results of a semi-quantitative assessment of the accumulation of compounds in cell lines are shown in FIG. 16. The fluorescence intensity of cells is significantly influenced by a ligand attached to chlorin-e ₆ .

It was shown that the compounds under study accumulate more in cells of a non-tumor nature (CHO). This may indicate a nonspecific accumulation of conjugates in cells — penetration by diffusion. However, this issue requires further study.

Analysis of the photodynamic activity of compounds in relation to cells of tumor and non-tumor nature.

Photodynamic activity was evaluated in vitro. For this, cells were planted on a 96-well plate in the amount of 4 × 103 per well and incubated at 37 ° C and an atmosphere of 5% CO ₂ for 24 hours. Then the nutrient medium was replaced with 200 μl of serum-free medium with a photosensitizer in various concentrations and the cells were incubated for 4 hours. After the end of the incubation, the medium with the compound in the plate wells was replaced with an appropriate growth nutrient medium. Next, the cells were irradiated using a LED emitter (Shilyagin et al., 2014) under thermostatically controlled conditions (37 ° C) at TermoStat plus (Eppendorf, Germany).

The radiation dose was 20 J / cm ² at a wavelength of 670 nm, the light output power was 32 mW / cm ² , the exposure time was 10 minutes and 25 seconds. The viability of the cell culture was evaluated 24 hours after irradiation with the MTT test.

MTT test.

To form a water-insoluble colored formazan crystals, MTT reagent (Alfa Aesar, United Kingdom) was introduced into the growth medium at a final concentration of 0.5 mg / ml and the cells were incubated for 4 hours. After the incubation medium was taken, 200 μl of dimethyl sulfoxide (DMSO) was added. The optical density of the solutions obtained in the well was measured on a Synergy MX plate spectrophotometer (BioTek, USA) at a wavelength of 570 nm. Cell viability was assessed by the ratio of the optical density of the formazan solution in each sample to the control (cells without a photosensitizer). The relative viability of the cells was calculated by the formula:

OPO is the averaged optical density in the wells with treated cells (minus the “blank”); OPK is the averaged optical density in the wells with untreated cells (minus the “blank”).

According to the obtained results, dose-response curves were constructed and the IC50 value for each compound was calculated (photosensitizer concentration reducing cell viability by 50%).

It was shown that all the compounds under study can cause photoinduced cell death, however, photodynamic activity differed significantly depending on the attached ligand.

Figures 16 and 17 show the graphs of the viability of A431 and CHO cells versus the concentrations of compounds 15, 17, 18, 23 in the dark and under illumination.

For compounds 15, 17, 18 (with carbohydrate ligands), dark toxicity was observed at concentrations of 15–35 μM for A431 cells and 80–90 μM for CHO cells. When irradiated at a dose of 20 J / cm2, the toxicity increased significantly. IC50 (the concentration of compounds at which there was a decrease in culture viability of 50%) did not exceed 2.5 μM in all cases (Fig. 17).

When added to compound 18 (with digalactosyl) as the second ligand, vandetanib, changes in the cytotoxicity of the compound are observed (Fig. 18). Compound 23 increases photodynamic activity and decreases dark toxicity compared with compound 18. The values of the inhibitory concentration of IC50 for all studied compounds are presented in Table 2.

Compound 23 has the lowest dark toxicity. The maximum light toxicity is observed for compounds 17 and 23. Compound 23 has the optimal darkness / light ratio (chlorin-e ₆ zinc complex conjugate with digalactosyl and vandetanib).

The data presented show that the presence of a hydrophilic maltose derivative attached to the chlorin fragment in the proposed derivative provides sufficient water solubility, allows the compounds obtained to selectively accumulate in tumor cells, which ensures their selective delivery to the tumor tissue and reduces their systemic toxicity.

Claims (19)

1. Derived zinc metal complex chlorin-e ₆ General formula

2. Derived under item 1, characterized in that when

is a zinc complex bromide 13- (dimethylethylammonium) amide-17 (3) - (1 "- deoxy-β-D-galactopyranosyl-1" - (1 ', 2', 3'-triazole-4'-yl ) -methyl) amidchlorin-e ₆ General formula

3. Derived under item 1, characterized in that when

is a zinc complex bromide 13- (dimethylethylammonium) amide-17 (3) - (1 "- deoxy-β-D-glucopyranosyl-1" - (1 ', 2', 3'-triazole-4'-yl ) -methyl) amidchlorin-e ₆ General formula

4. Derived under item 1, characterized in that when

is a zinc complex of 13- (dimethylaminoethyl) amide-17 (3) - (1 "- deoxy-β-maltosyl-1" - (1 ', 2', 3'-triazole-4'-yl) methyl ) amidchlorin-e _{6 of the} general formula

5. The derivative according to claim. 1, characterized in that when

is a zinc complex of 13- (N- (2- (2- (2-N- (4 - ((4'-bromo-2'-fluorophenyl) amino) -6-methoxyquinazolin-7) -1-oxobutyrato-amino-ethoxy) ethoxy ) ethyl)) amide-17 (3) - (1 "- deoxy-β-maltosyl-1" - (1 ', 2', 3'-triazol-4'-yl) methyl) amidchlorin-e ₆ general formula

6. The use of the derivative according to claims. 1-5 as an agent for photodynamic therapy.

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