Application of anticancer transcript modifying agents and chemotherapy or radiotherapy

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to application of transcript modifying agents to prevent the genetic grading of malignant cells required to resist cell damage and their survival in chemotherapy or radiotherapy. There is applied a pharmaceutical composition containing hydralazine and valproic acid or its salt, e.g. magnesium valproate, in a pharmaceutically acceptable carrier to assist in cancer treatment along with chemotherapy or radiotherapy.

EFFECT: invention provides synergetic effect in cancer treatment by conventional therapeutic agents.

5 cl, 5 ex, 9 dwg

 

The SCOPE of the INVENTION

The present invention relates to the use of modifying transcript agents to assist in the treatment of zlokacestvennosti by preventing cancerous cells from the implementation of the genetic changes necessary to counteract cellular damage and survival with chemotherapy or radiotherapy, more specifically it refers to the combined use of modifying transcript agents, inhibiting the mechanism of DNA methylation and dezazetilirovanie histones, and more specifically it relates to the use in combination through a set for treatment, containing hydralazine and valproate acid, or any salt, such as magnesium valproate, to assist in the treatment of cancer with chemotherapy or radiotherapy.

PRIOR art

Cancer or malignancy can be classified as solid or hematopoietic. Examples of the first include, among others, cancer, such as breast cancer, head and neck, colon and rectum. Examples of hematological type include leukemia and lymphoma. DNA in the cell nucleus is located in the chromatin and has several levels of order. A composite unit of chromatin is the nucleosome, consisting of octameric nuclear proteins known as histones, to which DNA is wound. The location of the sludge is the packaging of DNA in the nucleus plays an important role in gene regulation. Covalent modifications of histones, such as acetylation, have a major role in the regulation of chromatin and gene expression (Cho KS, Etizondo LI, Boerkoel CF: Advances in chromatin remodeling and human disease. Curr Opin Genet Dev 2004; 14: 308-15).

Currently, cancer remains an important global health problem. According to the International Agency for research on cancer and the world health organization, the incidence of this disease increases significantly, the evaluation shows that in 2000, there was 10 million new cases, and in 20 years they will be 15 million. (Mignogna MD, Fedele S, Russo LL. The World Cancer Report and the burden of oral cancer. Eur J Cancer Prev. 2004; 13: 139-42). On the other hand, the survival of patients suffering from the most common types of cancer, such as lung cancer, prostate cancer and breast cancer in recent years has definitely improved. Five-year survival was 50% in 1974, and in the period from 1992 to 1999, it increased to 63% (Jemal A, Murray T, Ward E, Samuels A, Tiwari RC, Ghafoor A, Feuer EJ, Thun MJ. Cancer statistics, 2005. CA known breast J Clin. 2005; 55: 10-30).

Although progress in the forms of treatment allowed to obtain a slight advantage with respect to survival, the results are still far from being optimal. Currently, chemotherapy along with surgery and radiotherapy are still the main principles of treatment, as the vast majority of patients is now, cancer patients need this form of therapy.

Extensive knowledge gained in recent years in the field of molecular nature of cancer, provided an opportunity to develop new forms of therapy, aimed mainly to block the function of oncogenes or reactivation of expression of suppressive genes. Examples of such works is the use of monoclonal antibodies against some oncogenic receptors, such as EGFR, HER2, and so forth. In the case of suppressive genes, some works in the field of therapy are the use of recombinant adenovirus, covering the encoding gene for functional product p53 (Hermiston TW, Kirn DH. Genetically based therapeutics for cancer: similarities and contrasts with traditional drug discovery and development. Mot Ther. 2005; 11: 496-507).

In General, the authors of this invention can recall such a new form of cancer therapy, as directed on a single gene product, or therapy of a single gene. However, this approach has serious drawbacks, as the genome of malignant cells is very adaptable, and as the nature of cancer is a multistage, so there is no single genetic changes responsible for the development of malignant phenotype. This means that although the blocking or restitution or gene product can lead to important antitumor effect that is specified is to function effectively will not be supported, as definitely malignant cell will ultimately develop resistance to this therapy, because malignant cells will increase or decrease gene expression, and thus can adapt to the effects of the specified therapy (Ross JS, Schenkein DP, Pietrusko R, Rolfe M, Linette GP, Stec J, Stagliano NE, Ginsburg GS, Symmans WF, Pusztai L, Hortobagyi GN. Targeted therapies for cancer 2004. Am J Clin Pathol. 2004; 122: 598-609).

Currently, it is well known that malignant cells have multiple defects, namely mutations, deletions, duplications, amplifications, as well as epigenetic changes, the latter are stable functional changes mainly due to modifications of chromatin, of which the two most important changes are DNA methylation and histone acetylation. Epigenetic changes should have a certain character and act in full functional compliance to maintain "malignant homeostasis". This concept is very important because all the defects of malignant cells are not a simple addition, it is consistent with the fact that proteins encoded by genes that play multiple roles in complex networks and interactive functions, controlling positive and negative feedback. In addition, negotiationprocesses, which occurs when the generation of the tumor, the cell must be maintained in a stable state between positive and negative signals as oncogenic pathways and suppressive genes to ensure that the processes of proliferation and cell death occurred according to the dynamics of a malignant condition (Weinstein IB. Cancer. Addiction to oncogenes - the Achilles heat of cancer. Science. 2002; 297 (5578): 63-4).

In addition to the internal complexity of the global gene expression of malignant cells, the picture becomes much more complex when trying regulation of gene expression as a result of exogenous stimuli, in particular due to chemotherapy and or radiotherapy effect. Chemotherapy and radiotherapy cause instant changes of gene transcription, and these changes occur not only in those genes that are primarily responsible for the carcinogenic processes, but also in genes that are not directly involved, for example, involved in metabolism, transport and so on (Alaoui-Jamali MA, Dupre I, Qiang H. Prediction of drug sensitivity and drug resistance in cancer by transcriptional and proteomic profiling. Drug Resist Updat. 2004:7: 245-55). The most important aspect, however, is that only those cells that are able to have adequate transcriptional response to a harmful stimulus, are the only cells that will survive the damage. Yas is about, that the answer to "adapt" to the survival occurred, it is necessary that the epigenetic mechanisms regulating transcription, were untouched. Therefore, if a malignant cell is influenced by modifying transcript agents, transcription signal, necessary for survival, will not occur, and the cell may become irreversible functional changes or undergo apoptosis. Transcription of eukaryotic cells can be defined as the ability of the above-mentioned cells to Express biologically active proteins. Therefore, transcription is a highly regulated phenomenon. The process starts at the gene level and ends at the protein level and includes many stages, therefore, the transcription has several levels of regulation, which include the following: 1) the chromatin structure, which is the physical construction of DNA that includes the level of chromatin packaging, which defines the ability of regulatory proteins to contact gene regulatory and promotor regions, 2) control of transcription initiation, 3) transport transcript, 4) transcript processing and modification, 5) stability of the transcript, 6) initiation of translation, 7) post-translational modification and 8) transport and stability of the protein. (Archambault J, Friesen JD. Genetics of eukaryotic RNA polymerases I, I, and III. Environ Rev. 1993; 57: 703-24). Undoubtedly, the transcriptional effects will be more important in their effect on higher-level regulation of transcription. Thus, modifying the transcript agents by actions at the highest level regulation of transcription will have the greatest impact on overall gene expression.

Covalent modifications of histones, such as acetylation and DNA methylation, play a significant role in determining the extent of chromatin packaging and ultimately in determining the overall gene expression, resulting in agents that inhibit DNA methylation and dezazetilirovanie histone demonstrated the ability to significantly change the expression. Loss of methylation could reduce the number of protein complexes associated with methylated domains in some States, leading to decreased activity of discontinuties that would inhibit agents inhibiting discontinuation. Loss of complexes that suppress transcription, may help re-Association of gene promoters with complexes that activate transcription with histone-acetylase activity. It is known that the most common form of DNA methyltransferase (DNMT1) can directly contact discontinuation, and that the amino-end also has the ability to contact corepressors (Nakao M. Epigenetics: interaction of DNA methylation and chromatin. Gene. 2001; 278: 25-31; Robertson KD. DNA methylation and chromatin-unraveling the tangled web. Oncogene. 2002; 21: 5361-79).

Despite the fact that have been tested hundreds of potential antitumor agents, cancer treatment person still requires attention, with many methods of cancer treatment are only partially effective potential in relation to adverse effects on virtually all systems. There is therefore a need not only to have a more effective therapeutic alternatives, but also to have some more specific therapies, which allow you to attack malignant cells are more selective way at the level of gene transcription. For this reason, one of the objectives of the present invention is the provision of a composition to support cancer treatment based on changes of the transcriptome, by modifying the transcription of agents, such as hydralazine and magnesium valproate by which cells become unable to survive under the influence of harmful incentives induced by chemotherapy or radiotherapy.

Antihypertensive agent hydralazine is an inhibitor of DNA methylation, which was used in the experimental systems to gipomotilinemii T cell DNA, h is about making these self-reactive cells (Yung R, Chang S, Hemati N, Johnson K, Richardson B. Mechanisms of drug-induced lupus. IV. Comparison of procainamide and hydralazine with analogs in vitro and in vivo. Arthritis Rheum. 1997; 40: 1436-43). Recently it was shown that hydralazine causes demethylation of promoter region suppressive genes and induces their reactivation models in vitro and in vivo; and that reactivated gene products are functional (Segura, Trejo-Becerril C, Pérez E, Chavez A, Salazar AM, Lizano M,Reactivation of tumor suppressor genes by the cardiovascular drugs hydralazine and procainamide and their potential use in cancer therapy. Clin Cancer Res. 2003; 9: 1596-603).

Hydralazine has a direct inhibiting effects on the DNA methyltransferase and models in vitro, it was shown that two atoms of nitrogen molecules interact with amino acids Lys162 and Arg240 active site of the enzyme responsible for the ability to demethylation and reactivation function suppressive genes (Angeles HER, Vazquez-Valadez, VH, Vasquez-Valadez Oh, Velazquez-Sanchez AM, Ramirez A, Martinez L, Diaz-Barriga S, Romero-Rojas A, Cabrera G,Duenas - Gonzalez A: Computational studies of 1-hydrazinophthalazine (Hydralazine) as antineoplasic agent. Docking studies on methyltransferase. Letters Drug Design Discovery 2005; 4:282-286).

Valproic acid or its salts, as in the case of magnesium valproate, also known as VPA, 2-propylpentanoic acid, a drug, which for many years was used as anti-convulsants good, as was shown by the profile of the traffic signals is (Perucca E: Pharmacological and therapeutic properties of valproate: a summary after 35 years of clinical experience. CNS Drugs 2002, 16: 695-714). Recently, it was shown that this drug is an inhibitor discontinuation. Inhibiting enzymes are enzymes of class 1 and II of the family except discontinuation 6 and 10. Hyperacetylation histone NC and H4, observed in vitro and in vivo, accompanied by the enzyme inhibitory effect of this family of enzymes. This action on the histones has a significant effect on the induction of differentiation, induction of apoptosis and inhibition of cell proliferation (Gurvich N, Tsygankova OHMS, Meinkoth JL, Klein PS: Histone deacetylase is a target of valproic acid-mediated cellular differentiation. Cancer Res 2004, 64:1079-1086).

For this reason, another object of the present invention is the provision of a kit for the treatment composed of hydralazine and valproic acid or any of its salts, such as magnesium valproate, which contributes to the commonly used anti-cancer therapy.

A BRIEF DESCRIPTION of GRAPHIC MATERIALS

On figa, B, C, D, E, F and G shown cytoxicity effect on the various lines of malignant cells treated modifying the transcript composition of the present invention.

On figa, and shows how modifying the transcript composition of the present invention causes a potentiation of the cytotoxic effect of representative chemotherapeutic the fir agents (cisplatin, doxorubicin and gemcitabine) on the line of malignant cells.

On figa shows the frequency of methylation of each gene according to the present invention.

On FIGU shows representative cases frequency of methylation according to the present invention.

On figs shows the correlation between percentage methylation and dose of hydralazine in accordance with the present invention.

On figa, b and C show typical cases of gene expression messenger in biopsies before and after processing.

Figure 5, 6 and 7 presents data on the activity discontinuties according to the present invention.

On Fig shows the inhibitory effect of the composition of the present invention to discontinuation.

Figure 9 shows the blocking effect on tumor growth in animals treated and untreated composition of the present invention.

DETAILED description of the INVENTION

According to the present invention proposed the use of a composition for the treatment to assist in treating zlokacestvennosti by applying modifying transcript compounds that complement the treatment with chemotherapy or radiotherapy.

Composition modifying transcript agents is a combination of hydralazine and valproic acid or any of its salts, such as valproate magni is, which contributes to chemotherapy, and chemotherapy can occur when the following compounds: hlorambuzila, cyclophosphamide, ifosfamide, mechlorethamine, melphalan, thiotepa, carmustine, lomustine, altretamine, dacarbazine and procarbazine, cisplatin, carboplatin and oxaliplatin, doxorubicin, daunorubicin, epirubicin, idarubitsina, mitomycin C, bleomycin, dactinomycin, retinoids, hormonal agents, vincristine, vinblastine, vindesine, vinorelbine, irinotecan, topotecan, etoposide, teniposide, paclitaxel, docetaxel, 5-fluorouracil, gemcitabine, methotrexate, interleukins, interferons, monoclonal antibodies, such as trastuzumab, cetuximab, Rituxan, mylotarg, and small inhibitory molecules, such as gefitinib, erlotinib and imatinib, but not limited to.

Composition and kit for treatment according to the present invention can be applied against tumors of several types, including, without limitation, breast cancer, ovary, uterus, skin, bone, prostate, liver, kidney, lung, brain, head and neck, gall bladder, pancreas, colon, rectum, parathyroid glands, thyroid gland, adrenal glands, stomach, kidney, pheochromocytoma, Wilms tumor, testicular cancer, neuroblastoma, sarcoma, acute and chronic leukemia, lymphoma, and m is eltoprazine syndromes.

The composition of the present invention can be administered orally or by any other by introducing into the drug containing 83 mg of hydralazine and valproate acid or its salt, such as magnesium valproate at a dose of 30 mg/kg (dry wt.), if the subject exhibits slow acetylation, or 182 mg of hydralazine and valproate acid or its salt, such as magnesium valproate at a dose of 30 mg/kg (dry wt.), if the subject has a quick acetylation. Both agent in any of their incarnations should be entered in the drug controlled release, starting with the introduction of the drug for 7 days prior to first dose of chemotherapy or the first session of radiotherapy to allow modification of the transcriptome in front of cytotoxic damage in these treatments.

EXAMPLES

APPLICATION EXAMPLE 1

To demonstrate that modifying the transcript composition, hydralazine and valproic acid or valproate magnesium have antitumor effects, used a number of malignant cell lines of cervical cancer, breast cancer, colon cancer, upper respiratory tract and digestive tract and sarcoma. The cells were placed in 96-well plates (Falcon Becton Dickinson, Franklin Lakes, NJ) with a density of 1.5-2.5×103cells/well in 0.1 ml complete medium. On the following day cells were treated with hydralazine at 10 μm and Valpro the volume of magnesium at 1 mm for 4 days. The next day we measured cell viability using the MTT-assay. Briefly, to each well was added 50 μl of MTT reagent (bromide 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium) in phosphate buffer solution. Viable cells with active mitochondria restore MTT deposited to purple compound - formazan, which is dissolved in DMSO (dimethyl sulfoxide) when adding 150 ál in each well. This was followed by spectrophotometric analysis of ELISA- (ELISA analysis) reader. All analyses were carried out in three repetitions. The cytotoxic effect of each treatment was expressed in percentage of cell viability relative to untreated control (% of control), which was defined as [(A570nmtreated cells)/(A570nmuntreated cells)]×100.

On figa, B, C, D, E, F and G shows that in all treated cell lines HeLa cell line carcinoma, cervical cancer, sarcoma NT, breast cancer MCF-7, epidermoid carcinoma of the larynx and oral cavity KB and NER, respectively; carcinoma of the colon SW480, epidermoid carcinoma of the oral cavity KB epidermoid carcinoma of the larynx NER,

D54 modifying the transcript composition resulted in significant cytotoxicity, changing in the range from 12.7%to 43.4%decrease in viability.

p> APPLICATION EXAMPLE 2

After demonstrating that modifying the transcript composition has any abscopal effects on the growth of malignant cell lines, doing research in regards to the increase in the composition of the cytotoxic effect of chemotherapeutic agents. For this purpose we have selected three drugs, which are typical representatives of their class of cisplatin alkylating agents, doxorubicin antibiotics, of gemcitabine antimetabolites. The cells were placed in 96-well plates (Falcon Becton Dickinson, Franklin Lakes, NJ) with a density of 1.5-2.5×103cells/well in 0.1 ml complete medium. On the following day cells were treated with a chemotherapeutic agent at a concentration shown on figa, b and C, and hydralazine at 10 μm and valproic acid or valproate magnesium at 1 mm. On the following day, the medium containing the drug was removed and added fresh hydralazine and magnesium valproate in the same concentrations for an additional 48 hours. On the next day (4th day) was measured cell viability using the MTT-assay. Briefly, to each well was added 50 μl of MTT reagent in phosphate buffer solution. Viable cells with active mitochondria restore MTT deposited to purple compound - formazan, which is soluble in DM what About when you add 150 ál in each well. This was followed by spectrophotometric analysis on a ELISA reader. All analyses were carried out in three repetitions. The cytotoxic effect of each treatment was expressed in percentage of cell viability relative to untreated control (% of control), which was defined as [(A570nmtreated cells)/(A570nmuntreated cells)]×100. On Figa, and it is shown that in all cases, the cytotoxicity of modifying the transcript of the composition and the chemotherapeutic agent is more significant. Under these conditions, the concentration of cisplatin 12 μm, which is the inhibitory concentration 50 (IC50), has reduced the viability of 37% in HeLa cells during the processing of modifying the transcript composition. A similar effect was also demonstrated for adriamycin and gemcitabine to reduce the viability of 27% and 37%, respectively.

APPLICATION EXAMPLE 3

To demonstrate that demetrious and revitalizing effect on transcription suppressive genes may be clinically achievable, was conducted phase 1 clinical trials to show the dose at which hydralazine may have demetrious and revitalizing effect transcription in patients with cancer. To this end, hydralazine was administered to groups of 4 patients in each group for 10 days at doses of: 1) 50 mg/day, 2 75 mg/day, 3) 100 mg/day and 4) 150 mg/day. Before treatment and on the 11th day took biopsies and peripheral blood samples. Analyzed the level of methylation before and after treatment for the promoters of the following genes: APC, MGMT; ER, GSTP1, DAPK, PARβ, FHIT and P16, as well as the expression level of their messengers by RT-PCR. Also assessed the level of methylation of the gene of the subject in relation to the parent decontamination n and genomic clone, which is usually considered methylated, and the total content of methylated cytokines in the genome. The toxicity of hydralazine was assessed using the scale of the National Cancer Institute of the USA (NCI CTC). Hydralazine was well-tolerated, and only the following undesirable effects have been reported: nausea, nausea (sickness), fatigue, headache and palpitations. As for genes, it was found that 70% of the analyzed samples (89 of 128) had at least one methylated gene in biopsies taken before treatment, 8 genes on each of 16 biopsies, patients, and all patients had their tumors at least one methylated gene. Individual analysis of each gene showed the following frequency of methylation: ARS 94%, ER 25%, FHIT 88%, GSTP1 88%, MGMT, 81%, P16 19%, RARβ 62% and DAPK 100%. In biopsies after treatment was detected variable frequency demethylation for each gene, changing on the AMAZONE from 15% in 2 out of 13 samples MGMT to 67% in 2 of 3 samples P16 gene. (Figa). Representative cases are presented on Figv. The correlation between the percentage of demethylation and dose of hydralazine was as follows: 50 mg of 40%, 75 mg, 52%, 100 mg, 43%, 150 mg, 32% (see figs).

Analysis of gene expression shows that 90% (116 out of 128) of the samples of the tumors expressed messenger in biopsy before and after treatment regardless of the level of methylation of the gene, therefore, they are not informative. 12 informative cases it was found that 9 of them had gene expression before treatment, which is methylated, but after treatment they were dimethylalanine and expressible gene. Representative cases are presented on figa, and in General, the frequency of gene reexpression summarized on fig.4D.

The above results indicate that hydralazine at doses between 50 and 150 mg is effective against changes in gene expression in patients with cancer (Zambrano P, Segura-Pacheco B, Pereez-Cardenas E, Cetina L, Revilla-Vazquez A, Taja-Chayeb L, Chavez - Blanco A, Angeles E, Cabrera G, Sandoval K, Trejo-Becerril C, Chanona-Vilchis J, Duenas-Gonzalez A. A phase I study of hydralazine to demethylate and reactivate the expression of tumor suppressor genes. BMC Cancer 2005; 5: 44).

APPLICATION EXAMPLE 4

To prove that valproic acid or its salt, such as magnesium valproate induces hyperacetylation of histones and inhibition activity discontinuties in tumor patients, cancer patients, was the province of the Deno another clinical trial, in which patients with cervical cancer were administered different doses of valproic acid or its salts, such as magnesium valproate. Twelve patients with a recent diagnosis of this cancer and without previous treatment received the following doses of magnesium valproate in groups of 4 patients. Group 1 - 20 mg/kg, group 2 - 30 mg/kg, group 3 - 40 mg/kg. Took a biopsy of the tumor and the blood sample before treatment and on the following day (6th day), as valproate acid or its salt, such as magnesium valproate was administered for 5 days in divided doses every 8 hours. Hyperacetylation of histone NC samples and histone H4 in tumor were analyzed using Western blotting analysis, activity discontinuties in extracts of nuclei tumors using colorimetric analysis, as well as the levels of valproic acid in serum. Also analyzed the gene expression level of P21 and CAR in biopsies after treatment. At the end of the treatment cycle was determined the toxicity of treatment. All patients received treatment; the average dose was 1890 mg/day with average values corresponding to the doses of 20, 30 and 40 mg/kg, 1245, 2000 and 2425 mg, respectively. Nine out of 12 patients were observed drowsiness degree 2. After treatment found hyperacetylation H3 and H4 nine and seven patients, respectively; and six of them have found hyperacetylation Wallpaper is of histones (positive and negative control, HeLa cells, treated or untreated trichostatin, respectively). Activity discontinuation decreased in 8 patients, while two patients that were statistically significant (two-sided t test p<0,0264) there was no change. (Positive and negative control, extracts of nuclei of HeLa cells, treated or untreated trichostatin), 5, 6 and 7. These results indicate that valproate magnesium used in doses is an effective and well-tolerated as an inhibitor discontinuation, which is reflected in the increase in expression of such genes like P21 (Fig).

APPLICATION EXAMPLE 5

After evaluating the ability of hydralazine and valproic acid or its salts, such as magnesium valproate, alter gene expression in tumor patients, the inventors conducted a study on the model of sarcoma in immunodeficient mice in relation to the following: increased whether the composition of these modifying transcript agents antitumor effect of chemotherapy. With this purpose, the investigated groups of 6 Nude mice-females that were injected with 6 million cells sarcoma cell lines NT. After the formation of tumors in animals regularly treated with hydralazine and valproic acid or its salt, such as magnesium valproate, in doses equivalent to those used is in relation to patients. There were formed the following groups of treatments: 1) control treated with saline, 2) treated once a week with adriamycin, 3) treated with the composition of hydralazine and valproate for 7 days followed the same week treatment with adriamycin. The results show that after 5 weeks of tumors in untreated animals amount to between 2 and 3 cm3and treatment with adriamycin leads to almost complete antitumor effect after 3 weeks after treatment, however, since that time the tumors are growing again, while in animals treated with composition, re-growth of tumors is blocked, as shown in Fig.9. The above suggests that modifying transcript combination prevents cell tumors from the implementation of the transcriptional changes necessary to restore the ability to grow. This phenomenon usually occurs in the treatment of cancer in patients whose observed that completely or almost completely antitumor response later often followed by relapse. Therefore, the composition of the present invention can induce delayed or complete remission of tumors.

The composition of the present invention may be included in a kit for treatment that should be administered orally or by any other way of introduction is in the drug containing 83 mg of hydralazine and valproate acid or its salt, such as magnesium valproate at a dose of 30 mg/kg (dry wt.), if the subject exhibits slow acetylation, and 182 mg of hydralazine and valproate acid or its salt, such as magnesium valproate at a dose of 30 mg/kg (dry wt.), if the subject has a quick acetylation. In order to avoid peaks in serum levels produced by hydralazine and valproic acid or its salt, such as magnesium valproate, and to reduce the side effects arising from their quick absorption, both agents should be entered in the drug controlled release.

Since the effect of hydralazine on the inhibition of methylation begins at least 48 hours after the administration, and the effect of valproic acid or its salts, such as magnesium valproate, transcription may be increased on the background of demethylation, the treatment composition preferably should begin seven days before the first dose of chemotherapy or radiotherapy to allow modification of the transcriptome in front of cytotoxic damage by chemotherapy or radiotherapy.

1. Pharmaceutical composition containing hydralazine and valproate acid or its salt, such as magnesium valproate, in a pharmaceutically acceptable carrier to assist in the treatment of cancer with chemotherapy Il is radiotherapy.

2. The pharmaceutical composition according to claim 1, containing from 50 to 100 mg of hydralazine and from 20 to 50 mg/kg valproic acid or its salts, such as magnesium valproate, in a pharmaceutically acceptable carrier for administration to subjects considered as having slow acetylation.

3. The pharmaceutical composition according to claim 1, containing from 150 to 200 mg of hydralazine and from 20 to 50 mg/kg valproic acid or its salts, such as magnesium valproate, in a pharmaceutically acceptable carrier for administration to subjects treated as having the rapid acetylation.

4. The pharmaceutical composition according to claim 1, where the chemotherapeutic agent include chlorambucil, cyclophosphamide, ifosfamide, mechlorethamine, melphalan, thiotepa, carmustin, lomustin, altretamin, dacarbazine and procarbazine, cisplatin, carboplatin and oxaliplatin, doxorubicin, daunorubicin, epirubicin, idarubitsin, mitomycin C, bleomycin, dactinomycin, retinoids, hormonal agents, vincristine, vinblastine, vindesine, vinorelbine, irinotecan, topotecan, etoposide, teniposide, paclitaxel, docetaxel, 5-fluorouracil, gemcitabine, methotrexate, interleukins, interferons, monoclonal antibodies such as trastuzumab, cetuximab, Rituxan, mylotarg, and inhibiting small molecules, such as gefitinib, erlotinib and imatinib, but not limited to.

5. The pharmaceutical composition according to claim 1, where the specified cancer is any solid tumor, including, without limitation, breast cancer, ovary, uterus, skin, bone, prostate, liver, kidney, lung, brain, head and neck, gall bladder, pancreas, colon, rectum, parathyroid glands, thyroid gland, adrenal glands, stomach, kidney, pheochromocytoma, Wilms tumor, testicular cancer, neuroblastoma, sarcoma, acute and chronic leukemia, lymphoma and myelodysplasia syndromes.



 

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24 cl, 20 tbl, 24 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel compounds selected from compounds of formulae Ia, lb and Ic, which have protein kinase activity on kinase selected from CDKs, Aurora, Jak2. Rock, CAMKI, FLT3, Tie2, TrkB, FGFR3 and KDR, abnormal activity of which is observed in pathological conditions such as nonmalignant and malignant proliferative diseases. In compounds of formulae , and : n equals 0 or 1, R1 is selected from a group comprising halogen, C1-C6alkyl, C1-C6alkoxy, halogen-substituted CpC6alkyl and halogen-substituted C1-C6alkoxy, R2 is selected from a group comprising phenyl, 6-member heteroaryl containing 1-2 nitrogen atoms in the heteroaryl ring as heteroatoms, and phenyl(C0-C4)alkyl, where the said phenyl and heteroaryl in R2 are optionally substituted with 1-3 radicals independently selected from a group comprising halogen, C1-C6alkyl, C1-C6alkoxy, halogen-substituted C1-C6alkyl, halogen-substituted C1-C6alkoxy, -S(O)0-2R5, -COOR5 and -NR5C(O)R6, where R5 is selected from C1-C6alkyl, and R6 is selected from phenyl, where the said phenyl in R6 is optionally substituted with 1-3 radicals independently selected from a group comprising C1-C6alkyl, C1-C6alkoxy, halogen-substituted C1-C6alkyl and halogen-substituted C1-C6alkoxy, X is selected from CR7 and N, where R7 is selected from hydrogen or C1-C6alkyl.

EFFECT: increased effectiveness of using the compounds.

7 cl, 3 dwg, 1 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to tetrahydropyridoindole derivatives of general formula , where R1, R2, R3 and R4 independently represent hydrogen; C1-C5alkyl, which can be optionally substituted and represents trifluoromethyl if C1-C5alkyl is substituted; C1-C3alkoxy or halogen, and R5 is C1-C6alkylcarbonyl, C1-C5alkylcarbamoyl, C1-C5alkoxycarbonyl, C2-C5alkenylcarbonyl, C3-C6cycloalkylcarbonyl, C3-C6cycloalkyl(C1-C3)alkylcarbonyl, C3-C6cycloalkylcarbamoyl, C3-C6cycloalkylthiocarbamoyl, phenylcarbonyl or phenyl(C1-C3)alkylcarbonyl, where the phenyl residue in these two groups contains one, two, three or four substitutes, independently selected from a group comprising C1-C4alkyl, C1-C3alkoxy, halogen, trifluoromethyl and trifluoromethoxy, monosubstituted with a C3-C6cycloalkyl group, or monosubstituted with a phenyl group which in turn is substituted with a C1-C3alkyl group; phenyl(C1-C3)alkoxycarbonyl, phenylcarbamoyl or phenylthiocarbamoyl (where these two groups are optionally independently monosubstituted with a C1-C5alkyl group or halogen atoms); phenyl(C1-C3)alkylcarbamoyl, phenyl(C1-C3)alkylthiocarbamoyl, biphenylcarbamoyl, naphthylcarbonyl, naphthyl(C1-C3)alkylcarbonyl or naphthylcarbamoyl (where the naphthyl residues in these three groups are optionally monosubstituted with substitutes independently selected from a group comprising C1-C3alkyl, C1-C3alkoxy and halogen); fluorenylcarbonyl, optionally substituted with an oxo group, fluorenyl(C1-C3)alkoxycarbonyl; or 5-9-member heteroarylcarbonyl groups containing one or two heteroatoms, independently selected from a group comprising oxygen, nitrogen and sulphur, where the said groups can be substituted with one or two groups independently selected from C1-C3alkyl and halogen, provided that if R1, R2, R3, R4 are hydrogen, R5 is not ethoxycarbonyl or tert-butoxycarbonyl, or salt thereof. The invention also relates to a pharmaceutical composition based on the compound of formula I and to use of the compound in preparing a medicinal agent.

EFFECT: obtaining novel tetrahydropyridoindole derivatives which have CRTH2 receptor antagonistic activity.

14 cl, 14 tbl, 171 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a free base (R)-3-[1-(2,6-dichloro-3-fluoro-phenyl)- ethoxy]-5-(1-piperidin-4-yl-1H-pyrazol-4-yl)-pyridin-2-ylamine in crystalline form, having a powder X-ray diffraction pattern with peaks at diffraction angles (2θ) 15.7±0.1, 17.3±0.1 and 19.7±0.1. The invention also relates to a pharmaceutical composition, to methods of treating cancer in mammals, as well as to a method of treating abnormal cell growth in mammals in need of such treatment.

EFFECT: obtaining an novel biologically active compound having the said inhibitory activity.

12 cl, 1 ex, 3 tbl, 2 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to novel compounds of formula

,

where the carbon atom denoted * is in R- or S-configuration; X is a concentrated bicyclic carbocycle or heterocycle selected from a group consisting of benzofuranyl, benzo[b]thiophenyl, benzoisothiazolyl, indazolyl, indolyl, benzooxazolyl, benzothiazolyl, indenyl, indanyl, dihydrobenzocycloheptenyl, naphthyl, tetrahydronaphthyl, quinolinyl, isoquinolinyl, quinoxalinyl, 2H-chromenyl, imidazo[1.2-a]pyridinyl, pyrazolo[1.5-a]pyridinyl, and condensed bicyclic carbocycle or condensed bicyclic heterocycle, optionally substituted with substitutes (1 to 4) which are defined below for R14; R1 is H, C1-C6-alkyl, C3-C6-cyclalkyl, C1-C3-alkyl, substituted OR11, -NR9R10 or -CN; R2 is H, C1-C6-alkyl, or gem-dimethyl; R3 is H, -OR11, C1-C6-alkyl or halogen; R4 is H, halogen, -OR11, -CN, C1-C6-alkyl, C1-C6-alkyl, substituted -NR9R10, C3-C6-cycloalkyl, substituted -NR9R10, C(O)R12; or R4 is morpholinyl, piperidinyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrrolyl, isoxazolyl, pyrrolidinyl, piperazinyl, 2-oxo-2H-pyridinyl, [1.2.4]triazolo[4.3-a]pyridinyl, 3-oxo-[1.2.4]triazolo[4.3-a]pyridinyl, quinoxalinyl, which are optionally substituted with substitutes (1 to 4) which are defined below for R14; R5 is H or C1-C6-alkyl; R6 is H, C1-C6-alkyl, or -OR11; R7 is H; R8 is H, -OR9, C1-C6-alkyl, -CN; R9 is H or C1-C4-alkyl; R10 is H or C1-C4-alkyl; or R9 and R10 taken together with the nitrogen atom to which they are bonded form morpholine; R11 is H, C1-C4-alkyl; R12 is C1-C6-alkyl; R14 in each case is independently selected from a substitute selected from a group consisting of halogen, -OR11, -NR11R12, C1-C6-alkyl, which is optionally substituted with 1-3 substitutes, in each case independently selected from a group consisting of C1-C3-alkyl, aryl; or to pharmaceutically acceptable salts thereof. The invention also relates to a pharmaceutical composition, to a method of obtaining formula (I) compounds, as well as to a method of treating disorders.

EFFECT: obtaining new biological active compounds having norepinephrine, dopamine and serotonin reuptake selective inhibitory activity.

90 cl, 162 ex, 2 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to a method of modulating expression of a target gene induced by β-catenin using an agent which increases linkage of p300 with β-catenin and reduces linkage of CBP with β-catenin, involving bringing a composition containing β-catenin, CBP and p300, where β-catenin is more likely linked to CBP than p300, into contact with an agent in an amount which is effective for changing the probability of linking β-catenin to CBP compared to p300, where the said agent is a compound with a structure selected from formula (I), or its stereoisomers: where A represents -(C=O)-, B represents -(CHR4)-, D represents -(C=O)-, E represents -(ZR6)-, G represents -(XR7)n-> W represents (C=O)NH-, X represents nitrogen or CH, Z represents CH, n = 0 or 1. Values of substitutes R1 and R2 are indicated in the formula of invention. The invention also relates to a composition for modulating expression of a target gene induced by β-catenin.

EFFECT: novel compounds have useful biological properties.

9 cl, 7 tbl, 30 dwg, 7 ex

FIELD: medicine.

SUBSTANCE: for treatment of pulmonary tuberculosis in HIV-infected patients carried out is anti-tuberculosis therapy according to standard schedules. Additionally since first day of treatment galavit is administered intramuscularly in dose 100 mg one time per 3 days. On the whole 15 injections are made. Addition of galavit to treatment, with given schedule and duration of introduction, produces normalising effect on imbalance of cellular and humoral immune response in patients with HIV-associated tuberculosis.

EFFECT: increase of treatment efficiency due to reduction of abacillation terms and closing of disintegration cavities, resolution of infiltrative changes in lungs.

2 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: inventions refers to medicine, namely to ophthalmology, and aims at treating the inflammatory exudative postoperative reactions following implantation of an intraocular lens. The conventional therapy is combined with intramuscular introduction of Galavit, the 1 and 2 day 0.2 g, and starting from the 4 day 0.1 g, 10 injections every second day.

EFFECT: method allows improving clinical effectiveness ensured by short-term reduction of the inflammatory reaction, improved visual acuity, shorter staying in hospital.

2 ex

FIELD: medicine.

SUBSTANCE: present invention concerns medical products, particularly pharmaceutical combination for development inhibition or treatment of proliferative disease which contains in therapeutically effective amounts (a) compound PTK787 and (b) epothilone derivative of formula , where A means O or NRN, where RN means hydrogen or lower alkyl, R means hydrogen or lower alkyl, and Z means oxygen or chain, for simultaneous, separate or consecutive application. Besides the invention concerns a pharmaceutical composition, application and commercial package thereof.

EFFECT: specified combination of active components ensures synergetic effect in treatment of proliferative diseases.

7 cl, 6 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a cyclic bioisostere of purine system derivatives, with general structural formula given below , where R = , Li, Na or K, R1 = -H, -NH2, -Br, -Cl, -OH, -COOH; A = -N- for B=-N=, Z = -CH-; A = -CH= for B = -N=, Z = -CH-; A = -CH= for B = -N=, Z = -N=; A = -CH= for B = -CH=, Z - -CH=; A = -CH= for B = -CH=, Z = -N=, except compounds in which A = -CH= for B = -CH=, Z = -CH=, R= Li, Na or K and R1= -NH2 in the 5th position of the benzo[d]-3H-pyridazine-1,4-dione nucleus, and its pharmacologically acceptable salts, with normalising effect on intracellular processes.

EFFECT: obtaining compounds which can be used for normalising intracellular processes in therapy of disorders, caused by intracellular acidosis and/or oxygen deficiency and/or excess formation of free radicals and/or excess formation of free radical forms of oxygen and/or high thrombocyte aggregation and/or erythrocytes and/or adverse effects and/or nitrergic cell mechanism disorder.

17 cl, 14 tbl, 15 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to a cyclic bioisostere of purine system derivatives, with general structural formula given below , where R = , Li, Na or K, R1 = -H, -NH2, -Br, -Cl, -OH, -COOH; A = -N- for B=-N=, Z = -CH-; A = -CH= for B = -N=, Z = -CH-; A = -CH= for B = -N=, Z = -N=; A = -CH= for B = -CH=, Z - -CH=; A = -CH= for B = -CH=, Z = -N=, except compounds in which A = -CH= for B = -CH=, Z = -CH=, R= Li, Na or K and R1= -NH2 in the 5th position of the benzo[d]-3H-pyridazine-1,4-dione nucleus, and its pharmacologically acceptable salts, with normalising effect on intracellular processes.

EFFECT: obtaining compounds which can be used for normalising intracellular processes in therapy of disorders, caused by intracellular acidosis and/or oxygen deficiency and/or excess formation of free radicals and/or excess formation of free radical forms of oxygen and/or high thrombocyte aggregation and/or erythrocytes and/or adverse effects and/or nitrergic cell mechanism disorder.

17 cl, 14 tbl, 15 dwg

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular to surgery, and concerns organism disintoxication in surgery of locally advanced hypernephroid cancer. With this purpose pre-operation and post-operation intramuscular introduction of sodium salt of 5-amino-1,2,3,4-tetrahydrophtalazine-1,4 dione (galavite) is carried out. Introduction is realised intramuscularly, daily in doze 0.1 g. Before operation 7 injections are made. After operation 5 injections are made, and then 10 injections every second day in the same doze.

EFFECT: due to claimed procedure of said medication introduction method ensures faster ending of catabolic phase of post-operation period with absence of side effect in patients with hypernephroid kidney cancer.

FIELD: medicine.

SUBSTANCE: inventions relates to medicine, in particular to surgery, and concerns disintoxication of organism at surgical treatment of patients with invasive cancer of urinary bladder. With this purpose pre-operation and post-operation intramuscular introduction of sodium salt of 5-amino-1,2,3,4-tetrahydroftalazine-1,4-dione (galavite) is carried out. Introduction is performed intramuscularly, daily in doze 0.1 g. Before operation 5 injections are made. After operation 5 injections are made, and then 10 injections every second day in the same doze.

EFFECT: due to claimed procedure of said medication introduction method ensures faster ending of catabolic phase of post-operation period with absence of side effect in patients with III-IV stage of disease.

FIELD: medicine.

SUBSTANCE: for treating patients suffering from chronic brucellosis, therapeutic complex includes an antibiotic, as well as Licopid and Tamerite. The preparations are introduced in therapeutic doses.

EFFECT: choice of specific preparations from the group of immunomodulators and antioxidants for specific infections provides long remission and prevention of unfavourable outcome of brucellosis.

3 ex, 3 tbl, 1 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: pharmaceutical composition contains a granulated component including multiple hardened melt granules of sugar alcohol containing salt of nonsteroidal anti-inflammatory drug (NSAID salt). The pharmaceutical composition is used for managing pain and/or inflammation and/or febrilily in coughing, cold, influenza, migraine, headache, rheumatic pain, arthritis pain, muscular pain and/or neuralgia.

EFFECT: according to the invention, the pharmaceutical composition contains minimum of tableting excipients, has improved fluidity, is less sticky, than NSAID salt itself and exhibits preferable tableting, disintegrating and dissolution properties.

39 cl, 3 dwg, 3 tbl, 29 ex

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