RU2557554C1 - Indolocarbazole derivative blocking tumour vasculogenic mimicry - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to indolocarbazole derivatives blocking tumour vasculogenic mimicry and applicable in medicine, of general formula I:

, wherein Gly is pentose and hexose residues, R represents an amino group of 6-amino-12-(α-L-arabinopyranosyl)indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7-dione; a formamido group of 6-formamido-12-(β-D-xylopyranosyl)indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7-dione; an oxylethylamino group of 6-(2-oxyethyl)amino-12-(α-L-arabinopyranosyl)indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7-dione; a cyanoethylamino group of 6-(2-cyanoethyl)amino-12-(β-D-xylopyranosyl)indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7-dione and 6-(2-cyanoethyl)amino-12-(β-D-galactopyranosyl)indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7-dione; a picolinamido group of 6-picolinamido-12-(β-D-xylopyranosyl)indolo[2,3-a]pyrrolo[3,4-c]carbazole-5,7-dione.

EFFECT: there are presented new compounds with the cytotoxic and antiangiogenic properties active with respect to HCT-116 colon cancer and Mel Kor melanoma.

6 ex, 1 tbl, 3 dwg

Description

The invention relates to medicine and pharmaceutical chemistry and relates to derivatives of indolocarbazoles that block vasculogenic mimicry in a tumor.

The formation of vascular channels by aggressive tumor cells without the participation of endothelial cells (EC) and fibroblasts limited by the basement membrane is called vasculogenic mimicry (VM). The formation of such structures is a unique ability of cells with a high-grade cell phenotype. It is believed that the formation of a network of such channels within the tumor can support homeostasis and prevent early necrosis within the tumor (Hendrix, M.J. et al. 2003, Oncogene, 22, 3070-3075). Clinical trials of drugs potentially aimed at lowering blood supply to the tumor revealed that antiangiogenic therapy of the tumor is not always effective (Azam, F. Et al. 2010, Eur J Cancer, 46 (8), 1323-1332). One of the reasons for the survival of tumor cells may be the heterogeneity of blood vessels: the formation of vessels in tumors occurs against the background of uncontrolled mitogenic stimulation and altered extracellular matrix. This leads to the development of inferior vessels, which often have a disturbed endothelial lining. The endothelium may be replaced by tumor cells or absent in the vessels of the tumor. The phenomenon of VM explains the insensitivity of many malignant tumors to antiangiogenic therapy. Earlier in vitro experiments, it was revealed and confirmed in vivo that the protein kinase C (PKC) inhibitor Ro32-0234, which is a derivative of indolocarbazole, dose-dependently reduces the density of vascular channels formed by tumor cells (A. Vartanian et al. Melanoma Res, 2011, 21 (2), 91-98).

The task of the invention is the creation of new derivatives of indolocarbazoles blocking BM in the tumor.

The problem is solved by the fact that N-glycosides of indolocarbazoles are proposed, which are maleimide indolo [2,3-a] carbazole, in which one of the indole atoms is linked by a glycosidic bond to the residues of various monosaccharides and contains various substituents at the imide nitrogen atom of the general formula I:

where Gly are the residues of pentoses and hexoses, R represents the amino group - 6-amino-12- (α-L-arabinopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione compound (1); formamido group - 6-formamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione, compound (2); hydroxyethylamino group - 6- (2-cyanoethyl) amino-12- (β-D-galactopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione, compound (3) , cyanoethylamino group - 6- (2-cyanoethyl) amino-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione, compounds (4 ) and 6- (2-hydroxyethyl) amino-12- (α-L-arabinopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione, compound (5);

picolinoamido group - 6-picolinamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione, compound (6).

To obtain compounds (1-6), indolo [2,3-a] furano [3,4-c] carbazole-5,7-dione N-glycosides were used as starting materials, which were condensed with hydrazine hydrate and various hydrazine derivatives, with subsequent removal of the protective groups of the carbohydrate fragment synthesized by the method described in the patent of the Russian Federation No. 2427585.

The resulting compounds were characterized by ¹ H-NMR and high resolution mass spectra. ¹ H-NMR spectra were recorded on a Bruker DRX-500 instrument with software, tetramethylsilane as the internal standard, and chemical shifts δ are given in parts per million (ppm). When describing the waveform, the following abbreviations are given: c - singlet, d - doublet, t - triplet, m - multiplet, dd - doublet of doublets, ears. from. - widened singlet; the values of the spin – spin interaction constants J are given in hertz. Mass spectra were recorded on a Finnigan MAT 8430 mass spectrometer with an SS-300 data processing system at an accelerating voltage of 3 kV, ionizing electron energy of 70 eV, ion source temperature of 250 ° C, sample evaporation temperature of 170-250 ° C, using a direct entering a substance into the ionization region. The values of m / z (the ratio of particle mass to charge) are given. The progress of the reactions was monitored by thin layer chromatography (TLC). TLC was performed on silofole.

Preparation of 6-amino-12- (α-L-arabinopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione, compound (1)

K 550 mg (0.9 mmol) 13-formyl-12- (2,3,4-tri-O-acetyl-α-L-arabinopyranosyl) indolo [2,3-a] furano [3,4-c] carbazole- 5,7-dione [RF patent No. 2427585] added 3.2 ml (65.25 mmol) of hydrazine hydrate, the mixture was kept at 50 ° C for two hours, cooled to a temperature of 22 ° C, poured into water. The precipitate was filtered off, washed with 10% alcohol. The yield of 6-amino-12- (α-L-arabino-pyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione was 370 mg (89%).

Mass spectrum: M ⁺ 457 m / z.

¹ H-NMR (DMSO-d ₆ , δ ppm): 12.15 (s, 1H, NH-indole), 9.23 (d, 1H, Ar), 9.14 (d, 1H, Ar), 7.94 (d, 1H, Ar), 7.70 (t, 1H, Ar), 7.62 (t, 1H, Ar), 7.59 (t, 1H, Ar), 7.41 (t, 1H, Ar), 7.37 (t, 1H, Ar ), 6.11 (d, 1H, H1 ′, J _{1′2 ′} 9.1), 6.86, 5.38, 5.24 (3 s, 3H, 2 ′, 3 ′, 4′-OH), 4.99 (2H, s, ΝΗ ₂ ), 4.20-4.05 (m, 3H, H2 ′, H3 ′, H5 ″), 4.25 (m, 1H, H5 ′), 3.85 (m, 1H, H4 ′).

Preparation of 6-formamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione, compound (2)

Mixture consisting of 100 mg (0.17 mmol) of 12- (2,3,4-tri-O-acetyl-β-D-xylopyranosyl) indolo [2,3-a] furano [3,4-c] carbazole -5,7-dione and 80 mg (1.33 mmol) of formyl hydrazide in 1.5 ml of dimethyl foramide, stirred at 75 ° C for 30 min. The solvent was evaporated. 10 ml of water was added to the residue. The precipitate was filtered off, washed with water (10 ml), dried, and 65 mg of acetylated 6-formamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5 was obtained, 7-dione.

To 65 mg (0.1 mmol) of acetylated 6-formamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione in 2 ml of absolute methanol was added 0.3 ml of a 0.1 N solution of sodium methylate in methanol. The reaction mixture was stirred at 22 ° C for 30 minutes, 1.0 ml of water was added. The precipitate was filtered off, washed with water (10 ml), and dried.

The yield of 6-formamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione (6) 26.0 mg (50%).

¹ H-NMR ((CD ₃ ) ₂ CO): 10.62 (s, 1H, NH), 9.88 (s, 1H, NH indole), 9.27 (d, 1H, Ar), 9.20 (d, 1H, Ar), 7.95 (d, 1H, Ar), 7.81 (d, 1H, Ar), 7.58 (t, 1H, Ar), 7.55 (t, 1H, Ar), 7.38 (t, 1H, Ar), 7.36 (t, 1H , Ar), 6.24 (broad d., 1H, H1 ′), 4.70-3.75 (m, 5H, H2 ′, H3 ′, H4 ′, H5 ′, H5 ″).

Mass spectrum: M ⁺ 457.

Preparation of 6- {2-cyanoethyl) amino-12- (β-D-galactopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione compound, compound (3)

Mixture consisting of 100 mg (0.16 mmol) of 12- (2,3,43,5-tetra-O-acetyl-β-D-galactopyranosyl) indolo [2,3-a] furano [3,4-c ] carbazole-5,7-dione and 40 mg and 40 mg (0.47 mmol) of 2-cyanoethylhydrazine in 1.5 ml of dimethylformide were stirred at 75 ° C for 1 hour. The solvent was evaporated. 10 ml of water was added to the residue. The precipitate was filtered off, washed with water (10 ml), dried, and 70 mg of acetylated 6- (2-cyanoethyl) amino-12- (β-D-galactopyranosyl) indolo [2,3-a] pyrrolo [3,4-c ] carbazole-5,7-dione.

To 70 mg (0.1 mmol) of acetylated 6- (2-cyanoethyl) amino-12- (β-D-galactopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7- dion in 2 ml of absolute methanol was added 0.5 ml of 0.1 N sodium methylate solution. The reaction mixture was stirred at 22 ° C for 30 minutes. The precipitate was filtered off, washed with water (10 ml), and dried. The yield of 6- (2-cyanoethyl) amino-12- (β-D-galactopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione was 29 mg (54%).

¹ H-NMR (DMSO-d ₆ ): 12.17 (s, 1H, NH), 11.07 (s, 1H, NH indole), 9.20 (d, 1H, Ar), 9.09 (d, 1H, Ar), 7.95 ( d, 1H, Ar), 7.70 (d, 1H, Ar), 7.59 (m, 2H, Ar), 7.37 (m, 2H, Ar), 6.18 (d, 1H, H1 ′, J _{1′2 ′} 7.0) , 6.82, 5.35, 5.21, 4.86 (4 broad s., 4H, 4-OH), 4.23 (m, 1H, H2 ′), 3.84 (m, 1H, H3 ′), 4.20-4.10 (m, 2H, H4 ′, H5 ′), 3.80-3.60 (m, 2H, H6 ′, H6 ″).

Mass spectrum: M ⁺ 487.

Preparation of 6- (2-cyanoethyl) amino-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione compound, compound (4)

6- (2-Cyanoethyl) amino-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione (4) was obtained from 12- (2 , 3,4-tri-O-acetyl-β-D-xylopyranosyl) indolo [2,3-a] furano [3,4-c] carbazole-5,7-dione is similar to the method for preparing compound (3).

The total yield of 6- (2-cyanoethyl) amino-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione (4) was 45 mg ( 33.4%).

% ¹ H-NMR ((CD ₃ ) ₂ CO): 10.62 (s, 1H, NH), 9.88 (s, 1H, NH indole), 9.27 (d, 1H, Ar), 9.20 (d, 1H, Ar), 7.95 (d, 1H, Ar), 7.81 (d, 1H, Ar), 7.58 (t, 1H, Ar), 7.55 (t, 1H, Ar), 7.38 (t, 1H, Ar), 7.36 (t, 1H , Ar), 6.24 (broad d., 1H, H1 ′), 4.70-3.75 (m, 5H, H2 ′, H3 ′, H4 ′, H5 ′, H5 ″).

Mass spectrum: M ⁺ 457.

Preparation of 6- (2-hydroxyethyl) amino-12- (α-L-arabinopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione compound, compound (5)

Mixture consisting of 150 mg (0.25 mmol) of 12- (2,3,4-tri-O-acetyl-α-L-arabinopyranosyl) indolo [2,3-a] furano [3,4-c] carbazole-5 , 7-dione and 137 mg (1.8 mmol) of 2-hydroxyethylaminohydrosine in 2 ml of dimethylformamide were stirred at 22 ° C for 16 hours. The solvent was evaporated. 10 ml of water was added to the residue. The precipitate was filtered off, washed with water (10 ml), dried, and 90 mg of acetylated 6- (2-hydroxyethyl) amino-12- (α-L-arabinopyranosyl) indolo [2,3-a] pyrrolo [3,4-c ] carbazole-5,7-dione.

To 90 mg (0.14 mmol) of acetylated 6- (2-hydroxyethyl) amino-12- (α-L-arabinopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7- dion in 2 ml of absolute methanol was added 0.3 ml of a 0.1 N solution of sodium methylate in methanol. The reaction mixture was stirred at 22 ° C for 30 minutes. The precipitate was filtered off, washed with water (10 ml), and dried. The yield of 6- (2-hydroxyethyl) amino-12- (α-L-arabinopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione was 48 mg (66.3% )

¹ H-NMR (DMSO-d ₆ ): 12.07 (s, 1H, NH), 9.97 (s, 1H, NH indole), 9.19 (d, 1H, Ar), 9.09 (d, 1H, Ar), 7.89 ( d, 1H, Ar), 7.67 (d, 1H, Ar), 7.54 (m, 2H, Ar), 7.35 (m, 2H, Ar), 6.05 (br d, 1H, H1 ′, J _{1′2 ′} 9.0 ), 6.75, 5.26, 5.14 (3 broad.s., 3H, 3-OH), 4.22 (m, 1H, H2 ′), 4.30-4.10 (m, 3H, H3 ′, H5 ′, H5 ″), 3.89 (m, 1H, H4 ′).

Mass spectrum: M ⁺ 457.

Preparation of 6-picolinamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione compound, compound (6)

A mixture consisting of 105 mg (0.18 mmol) of 12- (2,3,4-tri-O-acetyl-β-D-xylopyranosyl) indolo [2,3-a] furano [3,4-c] carbazole 5.7-dione and 110 mg (0.8 mmol) of α-picolinohydrazide in 1.5 ml of dimethyl foramide were stirred at 75 ° C for 1.5 minutes. The solvent was evaporated. 10 ml of water was added to the residue. The precipitate was filtered off, washed with water (10 ml), dried, and 70 mg of acetylated 6-picolinamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5 was obtained, 7-dione.

To 70 mg (0.1 mmol) of acetylated 6-picalinamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione. in 2 ml of absolute methanol was added 0.3 ml of a 0.1 N solution of sodium methylate in methanol. The reaction mixture was stirred at 22 ° C for 30 minutes. The precipitate was filtered off, washed with water (10 ml), and dried. The yield of 6-picolinamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione (6) was 37 mg (64.4%).

¹ H-NMR ((CD ₃ ) ₂ CD): 10.62 (s, 1H, NH), 9.88 (s, 1H, NH indole), 9.27 (d, 1H, Ar), 9.20 (d, 1H, Ar), 7.95 (d, 1H, Ar), 7.81 (d, 1H, Ar), 7.58 (t, 1H, Ar), 7.55 (t, 1H, Ar), 7.38 (t, 1H, Ar), 7.36 (t, 1H , Ar), 6.24 (broad d., 1H, H1 ′), 4.70-3.75 (m, 5H, H2 ′, H3 ′, H4 ′, H5 ′, H5 ″).

Mass spectrum: M ⁺ 457.

In vitro study of the cytotoxic activity of compounds (1-6)

A study of the cytotoxic activity of compounds (1-6) was carried out on HCT-116 human colon cancer cells and on Mel Kor metastatic melanoma cells,

Mel Cher, obtained from the tumor material of patients with disseminated melanoma at the N. N. N. Blokhina RAMS

Tumor cells were grown on RPMI 1640 medium containing 10% fetal calf serum at 5% CO ₂ and a temperature of 37 ° C. The cytotoxic activity of compounds (1-6) was evaluated using a standard MTT test using the MTT reagent (3,4,5-dimethylthiazol2-yl-2,5-diphenyltetrazolium bromide) based on the ability of dehydrogenases of living metabolically active cells to restore MTT- reagent to blue insoluble crystals of formazan [Guide to the experimental (preclinical) study of new pharmacological substances, M., 2005, p. 649-651].

Studies were performed on 96-well microplates. Tumor cells were scattered in 180 μl of culture medium and grown for 24 hours, then compounds (1-6) were added in concentrations from 0.01 to 100 μM in a volume of 20 μl. The incubation time of compounds (1-6) with tumor cells was 48 hours. Control was samples of cells to which compounds (1-6) were not added. After 48 hours, MTT reagent was added to the cells at a final concentration of 0.5 mg / ml. The cells were incubated for another 4 hours, then the cells were besieged by centrifugation of the plates at 1000 rpm for 2-3 minutes, then the medium was taken and 200 μl of DMSO was added to the cells to dissolve formazan. Tumor cells were resuspended and incubated for 10 min at 37 ° C, after which the optical density of the formazan solution was immediately determined on an Uniplan enzyme-linked immunosorbent analyzer at 540 nm using DMSO as a zero control.

The percentage of living cells was calculated by the formula:

where N ₀ is the percentage of living cells;

Ν ₁ is the average optical density of the wells containing cells and the test sample;

N ₂ is the average optical density of control wells containing only cells;

n is the optical density of the wells containing only the test sample.

For each compound, a dose-response relationship was plotted and ID _{50 was} determined. The measurement error in the experiments did not exceed 5%. In the study, the obtained compounds were used (1-6).

The invention is illustrated in the table and figures 1-3.

The table shows the cytotoxic activity of compounds (1-6) on human colon cancer cells HCT-116 and Mel Kor melanoma in vitro.

In FIG. 1 (A, B, C, D) presents the effect of compounds (1-6) on the formation of ATP by Mel Kor melanoma cells and HCT-116 colon cancer cells.

In FIG. 2 (A, B, C) presents a study of the effect of compounds (1-6) on the blocking of the migration of tumor cells of Mel Kor melanoma (test for “wound healing”).

In FIG. Figure 3 shows the effect of compound (6) on the formation of ATP by SVEC4-10 endothelial cells.

The table shows the values of the ID ₅₀ indicator of the concentration of compounds (1-6) required for 50% inhibition of tumor cells of colon cancer HCT-116. and melanomas Mel Kor. It was shown that the compound (2) possessed the highest cytotoxic activity.

In order to study the formation of the vascular network by tumor cells in vitro, migration and anti-angiogenic activity, the concentrations of compounds (1-6) were selected, during which incubation with them for 24 hours 90-95% of the tumor cells remained alive (non-cytotoxic doses). The use of non-cytotoxic doses of compounds (1-6) allowed us to avoid a false-positive result, in which the effect of blocking migration by 60% is associated with the death of tumor cells within 24 hours compared with the control.

Investigation of the effect of compounds (1-6) on the formation of vasculogenic mimicry in vitro.

The 24-well plate was coated with Matrigel (100 μl / well, density 8.4 mg / ml), incubated for 10 minutes in a laminar at room temperature, then 30 minutes in a CO ₂ incubator at 37 ° C until it polymerized. 200,000 Mel P, Mel Kor, and HCT-116 cells were incubated in 0.5 ml of complete RPMI 1640 medium supplemented with non-cytotoxic concentrations of compounds (1-6) for 30 minutes at 37 ° C. The cells were then transferred to Matrigel coated wells and incubated in a CO ₂ incubator for 20 to 24 hours. As a control, we used cells incubated in DMEM without addition of compounds (1-6). The length of the tubes and the number of contacts between the tubes in the PCA were evaluated. The maximum activity was possessed by compound (6).

In FIG. 1 (A, B, C and D) shows the effect of compound (6) on the formation of ATP by Mel Kor melanoma cells and HCT-116 colon cancer cells. In FIG. 1 (A and C) compound (6) at a concentration of 1 × 10 ^-7 480, 480 times lower _{than the} IC _50, blocked the formation of ATP by Mel Kor melanoma cells and HCT-116 colon cancer cells. In FIG. Figure 1 (B and D) shows Mel Kor melanoma and HCT-116 colon cancer cells on Matrigel without addition of compound (6), typical BM structures like “honeycombs” are visible. In FIG. 1 (B and D) shows the complete loss by tumor cells of Mel Kor melanoma and colon cancer HCT-116 the communication ability necessary for the formation of ATP, Matrigel had only small clusters of rounded cells after incubation with compound (6) at a concentration of 1 × 10 ^{- 7} M.

Study of the effect of compounds (1-6) on the migration activity of tumor cells

The formation of vascular-like structures (SPS) in 3D culture is characterized by a number of sequential events: the migration of tumor cells, the formation of cell-cell contacts, cell elongation and the formation of structures similar to honeycombs (honey-like comb).

Migration of melanoma cells and endothelial cells SVEC 4-10 was determined by the method of "wound healing" with some modifications. Mel P, Mel Kor melanoma cells in an amount of 3 × 10 ⁵ cells / ml were added to the wells of a 24-well plate in RPMI 1640 medium containing 10% fetal serum, 2 mM glutamine, an antibiotic, and incubated until a complete monolayer was formed. Then the monolayer was destroyed by scraping part of the cells in a straight line with a 200 μl tip. For 24 hours, cells were incubated in RPMI 1640 medium containing 10% fetal serum, 2 mM glutamine and non-cytotoxic doses (10 ^-6 to 10 ^-8 M) of compound (6). Tumor cells growing in complete RPMI 1640 medium and 10 ng / ml VEGFA served as a control. The results were evaluated by the change in the width of the migration surface after 24 hours of incubation after the addition of compounds (1-6) relative to the control.

In FIG. Figure 2 (A, B, C) shows that the addition of compounds (1-6) at non-cytotoxic concentrations did not affect the migration of Mel Kor melanoma cells. In FIG. 2A shows the migration of Mel Kor melanoma cells growing in a medium without embryonic serum; in FIG. 2B shows the migration of Mel Kor melanoma cells growing in complete medium supplemented with 10% fetal serum; in FIG. 2C shows the migration of cells growing in RPMI 1640 medium containing 10% fetal serum, 2 mM glutamine with the addition of non-cytotoxic doses (10 ^-7 M) of compound (6). It was shown that there were no differences in the width of the migration field compared with the control (arrows indicate the width of the migration field).

Study of the antiangiogenic activity of indolocarbazole derivatives

Compound (6) at a concentration of 10 ^-7 M, causing blockade of the formation of ATP by tumor cells, was tested for the ability to block tumor angiogenesis, i.e. the formation of blood vessels EC. For this purpose, we used the SVEC4-10 cell line obtained from the regional lymph nodes of C3H / HeJ mice (H-2 ^k ) and transformed with the SV40 virus large T-antigen from the ATCC collection (USA), which is widely used in world practice for screening pro- and antiangiogenic drugs. SVEC4-10 mouse ECs grow in culture without the addition of endogenous endothelial growth factors and extracellular matrix components, while maintaining the functional characteristics of normal microvascular ECs. SVEC-4-10 cells in an amount of 2 × 10 ⁵ cells / ml were incubated in DMEM with the addition of compound (6) in non-cytotoxic doses of 1 × 10 ^-8 M, which were active in blocking BM, for 30 minutes at 37 ° C and then transferred to Matrigel. In FIG. Figure 3 shows that compound (6) did not affect the formation of PCA EC of mouse SVEC4-10.

Claims (1)

Derivatives of indolocarbazoles blocking vasculogenous mimicry in the tumor,
general formula I:

where Gly are the residues of pentoses and hexoses, R represents the amino group of 6-amino-12- (α-L-arabinopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione; formamido group of 6-formamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione; hydroxyethylamino group of 6- (2-hydroxyethyl) amino-12- (α-L-arabinopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione; the cyanoethylamino group of 6- (2-cyanoethyl) amino-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione and 6- (2- cyanoethyl) amino-12- (β-D-galactopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione; the picolinoamido group of 6-picolinamido-12- (β-D-xylopyranosyl) indolo [2,3-a] pyrrolo [3,4-c] carbazole-5,7-dione.

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