RU2523387C1 - PHARMACEUTICAL COMPOSITION BASED ON 6-[(4-METHYL-1-1-PIPERAZINYL)METHYL]-INDOLO[1',7':1,2,3]PYRROLO[3',4':6,7]AZEPINO[4,5-b]INDOLE-1,3(2H,10 H)-DIONE AS ANTI-TUMOUR MEDICATION - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: claimed is a pharmaceutical composition, which contains as minimum one active component - a pharmaceutically acceptable salt of 6-[(4-methyl-1-1-piperazinyl)methyl]-indolo[1',7':1,2,3]pyrrolo[3',4':6,7]azepino[4,5-b]indole-1,3(2H,10 H)-dione of formula (I), in particular mesylate of 6-[(4-methyl-1-1-piperazinyl)methyl]-indolo[1',7':1,2,3]pyrrolo[3',4':6,7]azepino[4,5-b]indole-1,3(2H,10 H)-dione of formula (1)

and as minimum one auxiliary substance.

EFFECT: demonstrated suppression of growth of adenocarcinoma and melanoma tumour.

2 tbl

Description

The invention relates to the pharmaceutical industry and includes a pharmaceutical composition comprising a solution of a pharmaceutically acceptable salt of 6 - [(4-methyl-1-1-piperazinyl) methyl] -indolo [1 ', 7': 1,2,3] pyrrolo [3 ', 4': 6,7] azepino [4,5-b] indole-1,3 (2H, 10H) -dione of formula (I), in particular mesylate 6 - [(4-methyl-1-1 -piperazinyl) methyl] indolo [1 ', 7': 1,2,3] pyrrolo [3 ', 4': 6,7] azepino [4,5-b] indole-1,3 (2H, 10H) dione in a pharmaceutically acceptable polysorbate (tween), in particular polysorbate-60 (tween-60). The composition is used for the treatment of malignant tumors of humans or animals.

Formula I

where N = 1 or 2, and X represents the anion of any pharmacologically acceptable organic or inorganic acid.

State of the art

Tumor diseases are one of the main causes of death in Russia and around the world. Cancer cells are characterized by rapid division and invasiveness, i.e. ability to migrate and metastasize. In tumor cells, mechanisms of cell death (apoptosis) are defective, which otherwise could regulate their level, as well as proliferation and migration. In contrast, tumor cells have survival mechanisms that protect them from apoptosis. The Pim-1 enzyme (from its viral integration Moloney virus) protects cells from apoptosis; this mechanism contributes to the malignant transformation of cells [Moroy T., Grzeschiczek A., Petzold S., and Hartmann K. U.1993. Expression of Pim-1 transgene accelerates limphoproliferation and inhibits apoptosis in lpr / lpr mice. Proc. Nat. cad. Sci. 90 (22). 1073-4-10738.]. Pim-1 is overexpressed or activated in blood system tumors (hemoblastoses), as well as in prostate and mammary adenocarcinomas. For hemoblastoses, the role of Pim-1 in the development of B-cell non-Hodgkin lymphoma, diffuse B-cell lymphoma, Burkitt’s lymphoma and mantle-cell lymphoma [Sivertsen E.A., Galtrland E., Mu D., et al. 2006. Gain of chromosome 6p is an infrequent cause of increased PIM1 expression in B-cell non-Hodgkin's lymphomas. Leykaemia. 20. 539-42 .; Deutsch A., Aigelrransreiter A., Behan-Schmid C., Beham A., Linkesch W., Neumeister P.2005. Aberrant somatic hypermutayion in extranodal marginal zone B-cell lymphoma of MALT type. Blood. 106.125a-6a. Abstr 417. Rainio E. M., Ahlfors H., Carter K. L., et al. 2005. PIM kinases enchance EBNA2 activity. Virology. 333. 201-6. de Vos S., Krug U., Hofmann W.K., et al. 2003. Cell cycle alteration in the blastoid variant of mantle cell lymphoma (ML-BV) as detected by gene expression profiling of mantle cell lymphoma (MCL) and MCL-BV. Diagn Mol Pathol. 12. 35-43.18-21].

So, Pim-1 protein kinase is a pro-oncogenic protein that is important for the pathogenesis of tumors of the blood system and some solid tumors (prostate cancer, breast cancer). It follows that inhibition of Pim-1 activity is a necessary component of chemotherapy of these tumors.

Currently, several dozens of anticancer pharmacological drugs are used in clinical oncology, the effectiveness of most of which is insufficient, and the spectrum of oncological diseases treated with chemotherapy is significantly limited.

Known drug "Doxorubicin", an antitumor antibiotic of the anthracycline series. Doxorubicin has a high antitumor and anti-leukemia activity, has the ability to intercalate cell DNA. It has immunosuppressive activity. Doxorubicin is used for breast cancer, soft tissue sarcoma, osteogenic sarcoma, Jung's tumor, lung cancer, lymphosarcoma, ovarian cancer, squamous cell carcinoma of various localization, bladder cancer, Williams tumor, thyroid cancer, acute leukemia, lymphogranulomatosis. Enter only intravenously. The disadvantages of this tool is a depressing effect on blood formation, as well as a very high toxicity LD 50 13-15 mg / kg and cardiotoxic effect (Mashkovsky MD Medicines. Moscow. - 2001).

Known drug "Fluorouracil", which is one of the widely used anticancer drugs in clinical oncology and is used to treat ovarian cancer, breast cancer, liver cancer (hapatoma), melanoma, etc. (Safit JN, Bonavida B. // Cancer Res . - 1992. - Vol. 52. - P.6630). Like all cytostatics, 5-fluorouracil is very toxic - with its use, nausea, vomiting, inhibition of bone marrow hematopoiesis, toxic damage to the kidneys, etc. (Mashkovsky MD Medicines. Moscow. - 2001, p. 425).

The closest in structure is the substance described in the patent of the Russian Federation No. 2011108627 dated 03/09/2011, this patent also proposes the use of derivatives of 6- (4-methyl-1-1-piperazinyl) methyl-indolo [1 ', 7': 1,2,3] pyrrolo [3 ', 4': 6,7] azepino [4,5-b] indole-1,3 (2H, 10H) -dione of formula I as antitumor preparations, but specific compositions are not shown pharmaceutical compositions. The novelty lies in the fact that for the first time we proposed the composition, which allows to overcome the poor solubility of these substances in water and saline, which previously did not allow their use in the form of injectable forms for the treatment of tumors.

Disclosure of invention

This invention relates to a pharmaceutical composition, which is a solution of the active ingredient, a derivative of 6 - [(4-methyl-1-1-piperazinyl) methyl] -indolo [1 ', 7': 1,2,3] pyrrolo [3 ', 4 ': 6,7] azepino [4,5-b] indole-1,3 (2H, 10H) -dione of formula I, in particular mesylate 6 - [(4-methyl-1-1-piperazinyl) methyl] - indolo [1 ', 7': 1,2,3] pyrrolo [3 ', 4': 6,7] azepino [4,5-b] indole-1,3 (2H, 10H) -dione in a nonionic surface the active excipient is a pharmaceutically acceptable polysorbate (tween), in particular polysorbate-60 (tween-60), from which a pharmaceutical preparation can be easily prepared arat by dilution with aqueous solutions, such as saline.

Formula I

where X represents the anion of any pharmaceutically acceptable organic or inorganic acid, for example mesylate, and N = 1 or 2.

Polysorbate (Tween), intended for use in this invention, is, for example, Polysorbate-60 or other pharmaceutically acceptable polysorbate.

The term "Pharmaceutically acceptable" means a material that is used in the manufacture of a pharmaceutical composition and which is usually biologically or otherwise safe, non-toxic and acceptable both in veterinary medicine and in pharmaceuticals.

The formulation of the pharmaceutical composition of the present invention is a liquid, which, in particular, is an injectable solution, with particular preference for an injection solution (for example, for intravenous, subcutaneous, intramuscular, etc. injections) and intravenous fluids.

The pharmaceutical composition of the present invention can be used in the form of a liquid preparation for the treatment of malignant tumors.

An example of a method for producing a pharmaceutical composition:

50 mg of 6 - [(4-methyl-1-1-piperazinyl) methyl] -indolo [1 ', 7': 1,2,3] mesylate is added to 10 g of a 1% aqueous solution of Tween-60 at room temperature. pyrrolo [3 ', 4': 6,7] azepino [4,5-b] indole-1,3 (2H, 10H) dione and irradiated with ultrasound until completely dissolved.

Biological activity

Test substances were obtained by the methods described in the RF Patent No. 2011108627 dated 03/09/2011.

Antitumor activity of the dosage form of pimin in mice with subcutaneously inoculated adenocarcinoma of the mammary gland of mice AK 755

The study was conducted on BDF ₁ mice (C ₅₇ B1 × DBA ₂ ) females weighing 20-22 grams obtained from the Andreevka RAMA nursery. The animals were kept in vivarium of the FSBI NIINA RAMS. All animals had a veterinary passport - a certificate of animal health quality. The animals were kept in special cages of 10 individuals at an air temperature of 20-23 ° C and a relative humidity of 65% under artificial lighting and forced ventilation on a bed of wood shavings. For feeding animals, a standard industrial and certified extruded rodent feed with an established shelf life was used. Drinking water, placed in closed drinking bowls, animals received in unlimited quantities. After two weeks of quarantine, clinically healthy animals were introduced into the experiment. Groups of 10 mice each were formed.

Adenocarcinoma of the mammary gland of mice AK 755 (5th generation) was transplanted according to the standard technique subcutaneously in 1 million cells.

The drug was dissolved in 1% solution of TWEEN 60 in physiological sodium chloride solution and in the form of a 2% solution was administered to mice using a plastic syringe daily for 5 days intraperitoneally in single doses of 10 and 20 mg / kg, starting 48 hours after inoculation of the tumor.

,The condition and behavior of animals was monitored daily, the conditional volume of tumors was measured once a week (as a product of 3 tumor diameters). TPO% was calculated according to the generally accepted formula: $$\frac{V_k-V_o}{V_k}\times \mathrm{one\; hundred}\%$$

,

where: V _o is the average volume of tumors in the experimental group, V _k is the average volume of tumors in the control group.

Antitumor activity of the compounds are presented in table 1.

From the presented materials it can be seen that the drug, applied at a dose of 20 mg / kg daily for 5 days, shows a pronounced significant antitumor activity, which persists for 15 days after transplantation of the tumor. The drug, administered at a dose of 10 mg / kg, exhibits reliable antitumor activity only 11 days after tumor inoculation. By 15 days this effect is absent.

Antitumor activity of the dosage form of pimin in mice with subcutaneously inoculated melanoma of B-16 mice

The study was conducted on BDF ₁ mice (C ₅₇ B1 × DBA ₂ ) males weighing 20-22 grams obtained from the Andreevka RAMS nursery. The animals were kept in vivarium of the FSBI NIINA RAMS. All animals had a veterinary passport - a certificate of animal health quality. The animals were kept in special cages of 10 individuals at an air temperature of 20-23 ° C and a relative humidity of 65% under artificial lighting and forced ventilation on a bed of wood shavings. For feeding animals, a standard industrial and certified extruded rodent feed with an established shelf life was used. Drinking water, placed in closed drinking bowls, animals received in unlimited quantities.

After two weeks of quarantine, clinically healthy animals were introduced into the experiment. Groups of 10 mice each were formed.

The melanoma of B-16 mice (8th generation) was transplanted according to the standard procedure subcutaneously in 1 million cells.

The drug was dissolved in a 1% solution of TWEEN 60 in physiological sodium chloride solution and in the form of a 2% solution was administered to mice using a plastic syringe daily for 5 days intraperitoneally in single doses of 10; 20 and 30 mg / kg, starting 72 hours after tumor inoculation.

,The condition and behavior of animals was monitored daily, the conditional volume of tumors was measured once a week (as a product of 3 tumor diameters). TPO% was calculated according to the generally accepted formula: $$\frac{V_k-V_o}{V_k}\times \mathrm{one\; hundred}\%$$

,

where: V _o is the average volume of tumors in the experimental group, V _k is the average volume of tumors in the control group.

Antitumor activity of the compounds are presented in table 2.

With daily five-fold use in doses of 10 to 30 mg / kg, the drug in the dosage form exhibits pronounced antitumor activity against B-16 melanoma in mice, which lasts up to 21 days after transplantation of the tumor.

Table 1 Antitumor activity of the dosage form of pimin in mice with adenocarcinoma of the mammary gland of mice AK 755 TPO% Gr. No. A drug Dose (mg / kg) Day after inoculation eleven fifteen eighteen Tumor growth control one - - - - 2 Pimin in Leckform 10 × 5/24 69 41 37 3 Pimin in Leckform 20 × 5/24 74 86 43

table 2 Antitumor activity of the pharmaceutical composition in mice with B-16 melanoma TPO% Gr. No. A drug Dose (mg / kg) Day after inoculation eleven fifteen 21 Tumor growth control one - - - - 2 Pimin 10 × 5/24 89 72 68 3 Pimin 20 × 5/24 83 69 63 four Pimin 30 × 5/24 71 96 72

Claims (1)

A pharmaceutical composition for the treatment of malignant tumors containing at least one active ingredient, a pharmaceutically acceptable salt of 6 - [(4-methyl-1-1-piperazinyl) methyl] -indolo [1 ′, 7 ′: 1,2,3] pyrrolo [ 3 ′, 4 ′: 6,7] azepino [4,5-b] indole-1,3 (2H, 10H) -dione of the formula (I),

Formula I
where N = 1 or 2, and X represents the anion of any pharmacologically acceptable organic or inorganic acid, in particular mesylate 6 - [(4-methyl-1-1-piperazinyl) methyl] -indolo [1 ′, 7 ′: 1, 2,3] pyrrolo [3 ′, 4 ′: 6,7] azepino [4,5-b] indole-1,3 (2H, 10H) dione, and at least one excipient.