RU2441872C2 - Binuclear nitrosyl iron complexes with benzazeheterocyclic derivatives, ways of its production - Google Patents

Abstract

FIELD: pharmacology.

SUBSTANCE: invention refers to binuclear nitrosyl iron complexes with benzazeheterocyclic derivatives with general formula [Fe₂(SR)₂(NO)₄] where R is

And where X is NH, S, R¹ is lower alkyl. Also there is proposed method for its production, nitrogen monoxide donor, application as oncology drug and for production of oncology drug, pharmaceutical composition and set used for treatment of oncology diseased.

EFFECT: production of binuclear nitrosyl iron complex that is used as an oncology drug with increased effectiveness and decreased toxicity.

15 cl, 3 ex, 4 tbl, 3 dwg

Description

Technical field

The invention relates to binuclear nitrosyl iron complexes and can be used in medical practice to create new generation drugs for the treatment of cancer.

State of the art

In recent years, in order to create new-generation drugs used in the treatment of oncological diseases, an intensive search for antitumor drugs based on transition metal complexes with an improved spectrum of activity and reduced side effects compared with already used clinical drugs, for example cisplatin, sodium nitroprusside, etc. .P.

The interest in nitrosyl metal complexes has increased due to the open possibilities of their use as effective NO donors in medicine, in particular for the treatment of tumor diseases. The use of NO donors as a new class of antitumor agents is associated with the important role of NO in the growth of malignant tumors [Wink D., Vodovoz J., Cook J., Biochemistry, 1998, 63, 7, pp. 948-957]. Nitrogen monoxide has been shown to alter tumor cell apoptosis, p53 gene activity, and neoangiogenesis [Brune B., Scheneiderhan N., Nitric oxide evoked p53-accumulation and apoptosis, Toxicol Letters, 2003, 193, 2, pp.19-23], inhibits the activity of the key protein of repair of O6-methylguanine-DNA-methyltransferase of mammals [L. Liu, M. Xu-Welliver, S. Kanagula, H.E. Pegg, Inactivation and degradation of O6-alkylguanine-DNA alkyltransferase after reaction with nitric oxide., Canser. Res., 2002, 62, pp. 3037-3043].

The binding of NO to active sites of metal enzymes, in particular to non-heme iron-containing proteins, has been studied especially intensively [Ford PC, Lorkovic IM, Chem. Rev. 2002, 102, 993; Hoshino M., Laverman LE, Ford PC, Coord. Chem. Rev., 1999, 187, p. 75]. It has been established that one of the forms of natural NO reservoirs is nitrosyl iron complexes with thiol-containing ligands [Butler AR, Megson II, Chem. Rev. , 2002, 102, pp. 1155-1165].

Their synthetic models — Russen's red salt esters — have the composition [Fe ₂ (SR) ₂ (NO) ₄ ], where R = Et, t-Bu, (CH ₂ ) ₄ -CH ₃ , C ₆ H ₅ F, Ph [T. Thomas, JH Robertson, EG Cox, Acta. Crystalogr., 1958, 11, p. 599; C. Glidewell, ME Harman, MB Hursthouse, IL Johnson, M. Motevalli, J. Chem. Res., 1998, 212, p. 1676; RE Marsh, AL Spek, Acta. Crystalogr., Sect. B. Struct. Sci., 2001, 57, p. 800; C. Jinhua, M. Shaoping, H. Jinling, L. Jiaxi, Chinese J. Struct. Chem., 1983, 2, p. 263; TB Rauchfuss, TD Weatherill , Inorg. Chem., 1982, 21, pp. 827-830].

These binuclear diamagnetic seranitrosyl complexes with R = Alk generate NO upon thermal or photoactivation [J.L. Bourassa, P.C. Ford, Coord. Chem. Rev. 2000, 200-202, pp. 887-900] and can serve as new promising antitumor NO-donating agents.

Also known are binuclear paramagnetic seranitrosyl iron complexes with μ-N-C-S ligands, which are synthetic analogues of natural donors of NO-dinitrosyl iron complexes (DNIC) with cysteine, glutathione and other thiol-containing ligands of small molecular weight [O.A. Rakova, N.A. Sanina, S.M. Aldoshin, N.A. Goncharova, G.V.Shilov, Yu.M. Shulga, N.S. Ovanesyan, Inorganic Chemistry Communications 2003, 6, 145-148; A. F. Vanin, N. A. Sanina, V. A. Serezhenkov, D. Sh. Burbaev, V.I. Lozinsky and S.M. Aldoshin, Nitric oxide: biology & chemistry 2007, 16, 82-93].

However, the currently known synthetic NO donors of various classes are not used as therapeutic agents for the treatment of cancer, but are used only to enhance (to varying degrees depending on the chemical nature) the effects of existing chemotherapeutic agents or radiotherapy [Wink D., Vodovoz J. Cook J. Biochemistry 1998, 63, 7, pp. 948-957; N.P. Konovalova, S.A. Goncharova, L.M. Volkova, T.A. Raevskaya, L.T. Eremenko, A.M. Korolev, Nitric Oxide: Biology and Chemistry, 2003, 8, pp. 59-64; Yang, W., Rogers P.A., Ding H., J. Biol. Chem., 2002, 277, pp. 12868-12873; O. Siri, A. Tabard, P. Pullumbi, R. Guilard, Inorg. Chim. Acta 2003, 350, p. 633; J.L. Burgaud, E. Jngini, Del Soldato P. Ann, N.Y. Acad. Sci. 2002, 962, p. 360; T.I. Karu, L.V. Pyatibrat, G.S. Kalendo, Toxicology Letters, 2001, 121, p. 57].

In the work of A. Janczyk et al. Nitric Oxide, 2004, 10, 1, pp. 42-50, the direct cytotoxic effect of the nitrosyl iron complex Na [Fe ₄ S ₃ (NO) ₇ ] on human and mouse melanoma cells was studied. However, this nitrosyl iron complex generates NO during photoactivation (even in the dark), and also cannot be used as an antitumor drug due to its high toxicity to normal cells.

Thus, there is a need for other antitumor agents with increased efficacy and reduced toxicity.

The objective of the present invention is to expand the arsenal of antitumor agents and the creation of antitumor drugs based on transition metal complexes with an improved activity spectrum and reduced side effects, in particular, medicines based on binuclear nitrosyl iron complexes with benzazheterocyclic derivatives acting as NO donors with increased activity and reduced toxicity.

SUMMARY OF THE INVENTION

In one aspect, the invention relates to novel binuclear nitrosyl iron complexes with benzazheterocyclic derivatives of the formula [Fe ₂ (SR) ₂ (NO) ₄ ], where R is

where X represents NH, S or O, R ¹ represents lower alkyl.

In another aspect, the invention relates to a method for producing binuclear nitrosyl iron complexes with benzazheterocyclic derivatives of the formula [Fe ₂ (SR) ₂ (NO) ₄ ] by treating the thiosulfate nitrosyl iron complex with an appropriate benzazheterocyclic thiol in a stoichiometric ratio in the presence of a reducing agent in an alkaline medium. This method allows for the first time to obtain the claimed complexes in crystalline form.

In an additional aspect, the present invention relates to a donor of nitric oxide, which is a binuclear nitrosyl complex of iron with benzazheterocyclic derivatives, described above.

In a further aspect, the present invention relates to the use of binuclear nitrosyl iron complexes with benzazheterocyclic derivatives of the formula [Fe ₂ (SR) ₂ (NO) ₄ ] as an antitumor drug.

In an additional aspect, the present invention relates to the use of binuclear nitrosyl iron complexes with benzazheterocyclic derivatives of the formula [Fe ₂ (SR) ₂ (NO) ₄ ] for the preparation of antitumor drugs.

In a further aspect, the present invention relates to a pharmaceutical composition comprising an effective amount of a binuclear nitrosyl iron complex with benzazheterocyclic derivatives of the formula [Fe ₂ (SR) ₂ (NO) ₄ ] and a pharmaceutically acceptable carrier.

In a further aspect, the present invention relates to a kit used to treat cancer, comprising: (1) a pharmaceutical composition comprising a binuclear nitrosyl iron complex with benzazheterocyclic derivatives of the formula [Fe ₂ (SR) ₂ (NO) ₄ ], where R has the above meanings in sealed packaging; and (2) auxiliaries.

Description of figures:

Figure 1 shows the molecular structure of the complex (Fe ₂ S ₂ N ₈ C ₁₄ H ₁₀ O ₄ ) [complex I].

The complex has a centrosymmetric dimeric binuclear structure (Fig. 1). In the dimer, two tetrahedrally coordinated by two NO groups and two benzazheterocyclic thiol iron atoms are connected by a bridge: Fe-S-C-N-Fe. The complex contains two solvent molecules - acetone.

Figure 2 shows the crystal structure of the complex (Fe ₂ S ₂ N ₈ C ₁₄ H ₁₀ O ₄ ) [complex I].

Figure 3 shows the change in the difference absorption spectra over time during the interaction of complex I (Fe ₂ S ₂ N ₈ C ₁₄ H ₁₀ O ₄ ) with hemoglobin (Hb). The solvent is 0.05% phosphate buffer, pH 7.0, containing 3.3% dimethyl sulfoxide, 25 ° C.

DETAILED DESCRIPTION OF THE INVENTION

The new binuclear nitrosyl iron complexes with the benzazheterocyclic derivatives of the present invention have the general formula [Fe ₂ (SR) ₂ (NO) ₄ ], where R is

where X represents NH, S, R ¹ represents lower alkyl.

Preferably, R is benzimidazol-2-yl, 5-methyl-benzimidazol-2-yl or benzthiazol-2-yl.

The term “lower alkyl” as used herein means a straight or branched chain alkyl radical containing from 1 to 6 carbon atoms. Examples of such alkyl radicals include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl.

In the prior art there is no information about the claimed compounds and the method for their preparation.

A method of obtaining new binuclear nitrosyl iron complexes with benzazheterocyclic derivatives of the formula [Fe ₂ (SR) ₂ (NO) ₄ ], where R has the above meanings, is that the thiosulfate nitrosyl iron complex is bis (μ-thiosulfato-S) -bis ( dinitrosylferrate) (2-) sodium Na ₂ [Fe ₂ (S ₂ O ₃ ) ₂ (NO) ₄ ] (TNLC) is treated with the corresponding benzazheterocyclic thiol in a stoichiometric ratio in the presence of a reducing agent, and the process is carried out in an alkaline medium followed by isolation of the target product by known receptions.

Preferably, the process is carried out at room temperature, preferably at 18-25 ° C.

Preferably, the process is conducted in an oxygen free atmosphere.

As the benzazheterocyclic thiol, benzimidazole-2-thiol, 5-methylbenzimidazole-2-thiol or benzthiazole-2-thiol are preferably used.

Hydrogen, metal thiosulfates, for example Na ₂ S ₂ O ₃ · 5H ₂ O, hydrogen sulfide and aliphatic thiols are used as reducing agent in the method of the present invention.

The inventors have found that binuclear nitrosyl iron complexes with the benzazheterocyclic derivatives of the present invention are effective NO donors, generating nitrogen monoxide spontaneously when decomposed in proton media (such as water, blood and its components, physiological solutions, etc.), in the absence of chemo, photo, or enzymatic activation. Thus, the complexes of the present invention represent a new promising class of nitrogen monoxide donors — synthetic analogues of active sites of nitrosyl non-heme iron-sulfur proteins — natural NO depots. Therefore, the present invention in an additional aspect relates to nitric oxide donors, which are binuclear nitrosyl iron complexes with benzazheterocyclic derivatives described above.

Binuclear nitrosyl iron complexes with the benzazheterocyclic derivatives of the natural aromatic thiols of the present invention have antitumor activity against tumor cells in a mammal, in particular humans.

The present invention is further directed to the use of binuclear nitrosyl iron complexes with benzazheterocyclic derivatives of the formula [Fe ₂ (SR) ₂ (NO) ₄ ], where R has the above meanings, as an antitumor drug.

In particular, the compounds of the present invention can be used to treat the following oncological diseases: ovarian cancer, breast adenocarcinoma, B-16 melanoma, Lewis epidermoid carcinoma.

The iron complexes of the present invention are suitable for inhibiting tumor growth in mammals and are preferably administered in the form of a pharmaceutical composition comprising an effective antitumor amount of a compound of the present invention in combination with at least one pharmaceutically or pharmacologically acceptable carrier and / or excipient. A carrier, also known in the art as an excipient, excipient, excipient, additive or diluent, is any substance that is pharmaceutically inert, gives the appropriate consistency or form of the composition and does not impair the therapeutic efficacy of the antitumor compounds. The carrier is ″ pharmaceutically or pharmacologically acceptable ″ if it does not cause an adverse, allergic or other adverse reaction when administered to a mammal or human, respectively.

The present invention also relates to the use of binuclear nitrosyl iron complexes with benzazheterocyclic derivatives of the formula [Fe ₂ (SR) ₂ (NO) ₄ ], where R has the above meanings, to obtain an antitumor drug.

The present invention further provides pharmaceutical compositions comprising an effective amount of a binuclear nitrosyl iron complex with a benzazheterocyclic derivative of the formula [Fe ₂ (SR) ₂ (NO) ₄ ], wherein R is as defined above, and a pharmaceutically acceptable carrier.

Preferably, a proton-containing medium is used as a pharmaceutically acceptable carrier in the composition of the invention.

As a pharmaceutically acceptable carrier, a mixture of a proton-containing medium and dimethyl sulfoxide is also preferably used in the composition of the invention.

Preferably, water, physiological saline, and water-soluble biopolymers are used as the proton-containing medium.

Preferably, a binuclear nitrosyl iron complex with a benzazheterocyclic derivative is present in the composition in an amount of 50-100 μM.

Pharmaceutical compositions containing the antitumor compounds of the present invention can be prepared by any conventional method. The required formulation is selected depending on the chosen route of administration. Compositions according to the invention can be prepared for any route of administration so that the target tissue is accessible with this route of administration. Suitable routes of administration include, but are not limited to, oral, parenteral (e.g., intravenous, intraarterial, subcutaneous, rectal, intramuscular, intraorbital, intracapsular, intraspinal, intraperitoneal or intrasternal), local (nasal, percutaneous, intraocular, intraocular) into the small intestine, pulmonary, intralymphatic, intracavitary, vaginal, trasurethral, intradermal, auricular, intramammary, buccal, orthotopic, intratracheal, intrafocal, percutaneous, endoscopic, transmucosal, sublingual and intestinal injections.

Pharmaceutically acceptable carriers for use in the compositions of the present invention are well known to those skilled in the art and are selected depending on a number of factors: the particular antitumor compound used and its concentration, stability and expected bioavailability; the disease, disorder or condition of the person being treated with the composition; subject, his age, weight and general condition; and method of administration. Suitable carriers are readily identifiable by a person skilled in the art (J.G. Nairn in: Remington's Pharmaceutical Science (published by A. Gennaro), Mack Publishing Co., Easton, Pa, 1985, pp. 1492-1517).

The compositions are preferably prepared in the form of tablets, dispersible powders, pills, capsules, gelatin capsules, coated droplets, gels, liposomes, granules, solutions, suspensions, emulsions, syrups, elixirs, lozenges, dragees, lozenges, or any other dosage form, which can be administered orally.

Compositions of the present invention for oral administration comprise an effective antitumor amount of a compound of the invention in a pharmaceutically acceptable carrier. Suitable carriers for solid dosage forms include sugars, starches and other common substances including lactose, talc, sucrose, gelatin, carboxymethyl cellulose, agar, mannitol, sorbitol, calcium phosphate, calcium carbonate, sodium carbonate, kaolin, alginic acid, gum arabic, corn starch , potato starch, sodium saccharinate, magnesium carbonate, tragacanth, microcrystalline cellulose, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate and stearic acid. Further, such solid dosage forms may be uncoated or may be coated by known methods, for example, to delay disruption and absorption.

The antitumor compounds of the present invention are also preferably used for the preparation of a parenteral dosage form, for example, in the form of a dosage form for injection by intravenous, intraarterial, subcutaneous, rectal, intramuscular, intraorbital, intracapsular, intraspinal, intraperitoneal or intrasterial route. Compositions according to the invention for parenteral administration include an effective antitumor amount of an antitumor compound in a pharmaceutically acceptable carrier. Formulations suitable for parenteral administration include solutions, suspensions, dispersions, emulsions, or any other dosage form that can be administered parenterally. Techniques and compositions for preparing parenteral dosage forms are known in the art.

A minor amount of additional components well known in the pharmaceutical industry may be included in the compositions of the invention for various purposes. These components for the most part give properties that increase the retention time of the antitumor compound at the injection site, contribute to the stability of the composition, provide pH control, facilitate the incorporation of the antitumor compound into pharmaceutical compositions, and the like. Each of these components is individually present in an amount, preferably, less than about 15 wt.%, More preferably, less than about 5 wt.% And, most preferably, less than about 0.5 wt.% Based on the total weight of the composition. Some components, such as fillers or diluents, can be up to 90 wt.% Based on the total weight of the composition, as is well known in the technology of preparation of medicines. Such additives include cryoprotective components to prevent precipitation of the iron complex with thiophenol, surfactants, wetting or emulsifying agents (for example, lecithin, polysorbate-80, ^Tween® 80, pluronic-60, polyoxyethylene stearate), preservatives (for example, ethyl p- hydroxybenzoate), anti-microbial agents (e.g. benzyl alcohol, phenol, m-cresol, chlorobutanol, sorbic acid, thimerosal and paraben), pH adjusting agents or buffering agents (e.g. acids, bases, sodium acetate, sorbitan monolaurate), components for regulating osmolarity (e.g. glycerin), thickeners (e.g. aluminum monostearate, stearic acid, cetyl alcohol, stearyl alcohol, guar gum, methyl cellulose, hydroxypropyl cellulose, tristearin, cetolic esters) , pigments, dyes, flow agents, non-volatile silicones (e.g. cyclomethicone), clays (e.g. bentonites), adhesives, bulking agents, flavoring agents, sweeteners, adsorbents, nap fillers (e.g., water, saline, electrolyte solutions), binders (e.g., starches, such as corn starch, wheat starch, rice starch or potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, polymeric, polyvinyl chloride, polyvinyl ), disintegrating agents (e.g., starches such as corn starch, wheat starch, rice starch, potato starch or carboxymethyl starch, structured polyvinylpyrrolidone, agar, al gynic acid or its salt, such as sodium alginate, croscarmellose sodium or crospovidone), lubricants (e.g., silica, talc, stearic acid or its salts, such as magnesium stearate, or polyethylene glycol), coating agents (e.g., concentrated sugar solutions including gum arabic, talc, polyvinylpyrrolidone, carbopolgel, polyethylene glycol or titanium dioxide) and antioxidants (e.g. sodium metabisulfite, sodium bisulfite, sodium sulfite, dextrose, phenols and thiophenols).

According to a preferred embodiment, the pharmaceutical composition according to the invention comprises at least one non-aqueous pharmaceutically acceptable solvent and an antitumor compound having a solubility therein of at least about 10-60 mg / ml. Without regard to any particular theory, it is believed that the solubility of the antitumor compound in dimethyl sulfoxide can be directly related to its effectiveness. It is also preferred that the antitumor compound has an ID ₁₀₀ value (i.e., a drug concentration that causes 100% inhibition of colony formation) at least 4 times less than cisplatin when measured in accordance with the procedure described in the book "Experimental evaluation of anticancer drugs in the USSR and the USA", ed. Z.P. Sof'ina, A.B. Syrkina (USSR), A. Goldin, A. Klein (USA) M .: Medicine, 1979, p. 71-105.

The introduction of the dosage form in these ways can be continuous or periodic, depending, for example, on the physiological condition of the patient, on whether the purpose of the introduction of therapeutic or prophylactic, and other factors known or evaluated by the practitioner.

The dose and administration regimen of the pharmaceutical compositions of the invention can be easily determined by an oncologist. It is understood that the dose of antitumor compounds depends on the age, gender, state of health and weight of the recipient, the type of treatment carried out simultaneously, if any, the frequency of treatment and the nature of the desired effect. For any route of administration, the exact amount of the antitumor compound used, as well as the prescribed dose needed to achieve the beneficial effects described here, also depends, in particular, on factors such as the bioavailability of the antitumor compound, the disease of the person being treated, the desired therapeutic dose, and others factors that are obvious to the specialist.

The concentration of the antitumor compound in the liquid pharmaceutical composition is most preferably 50-100 μM. Relatively low concentrations are generally preferred, since the antitumor compound is most soluble at low concentrations.

Emulsions for parenteral administration can be prepared by dissolving the antitumor compound in any pharmaceutically acceptable solvent capable of dissolving the compound (e.g., DMSO, dichloromethane) to form a solution. An appropriate volume of carrier is added to the solution with stirring, which is an emulsion, such as an Liposyn II or Liposyn III emulsion, to form a pharmaceutically acceptable emulsion for parenteral administration to a patient. If desired, such emulsions can be prepared with or without a minimum Cremophor® solution.

Solutions for parenteral administration can be prepared by dissolving the antitumor compound in any pharmaceutically acceptable solvent capable of dissolving the compound (eg, DMSO) to form a solution. To the solution, with stirring, add the appropriate volume of the carrier, which is a proton-containing medium (saline solutions, sugar solutions, water-soluble polymers, proteins, electrolyte solutions) for parenteral administration to the patient.

For example, for liquid formulations, non-aqueous pharmaceutically acceptable polar solvents are usually used as a carrier, such as oils, alcohols, amides, esters, ethers, ketones, hydrocarbons and mixtures thereof, as well as water, saline solutions, dextrose solutions ( for example, DW5), electrolyte solutions, or any other proton pharmaceutically acceptable medium.

Suitable non-aqueous pharmaceutically acceptable polar solvents include, but are not limited to, amides (e.g. dimethylacetamide (DMA), dimethylacetamide benzyl benzoate, dimethylformamide, N- (β-hydroxyethyl) lactamide, N, N-dimethylacetamide, 2-pyrrolidinone, 1-methyl-2 -pyrrolidinone or polyvinylpyrrolidone); esters (e.g., acetic acid esters such as monoacetin, diacetin and triacetin; aliphatic or aromatic esters such as ethyl caprylate or ethyl octanoate, alkyl oleate, benzyl benzoate, benzyl acetate, dimethyl sulfoxide (DMSO), glycerol esters such as mono-di - or triglyceryl citrates or tartrates, ethyl benzoate, ethyl acetate, ethyl carbonate, ethyl lactate, ethyl oleate, sorbitan fatty acid esters, fatty acid esters of polyethylene glycol (PEG), glyceryl monostearate, glyceride esters such as mono, di or tri fatty acid esters, such as isopropyl myristate, fatty acid esters and PEG, such as PEG hydroxyoleate and PEG hydroxystearate, N-methylpyrrolidinone, pluronic-60, polyoxyethylene sorbitol and oleic acid polyesters such as poly ( oxyethylene) _30-60 sorbitol poly (oleate) _2-4 , poly (oxyethylene) _15-20 monooleate, poly (oxyethylene) _15-20 mono-12-hydroxystearate and poly (oxyethylene) _15-20 monoricinoleate; polyoxyethylene sorbitan esters such as polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monopoly limitate, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monostearate and polysorbate® 20, 40, 60 or 80 from ICI Americas, Wilmington, DE; polyvinylpyrrolidone; alkylene oxide-modified fatty acid esters such as hydrogenated polyoxy (40) castor oil and polyoxyethylene castor oils (for example, Cremophor® EL solution or Cremophor® RH 40 solution); fatty acid esters of a saccharide (i.e., a condensation product of a monosaccharide (e.g., pentoses, such as ribose, ribulose, arabinose, xylose, lyxose and xylulose; hexoses, such as glucose, fructose, galactose, mannose and sorbose; trioses, tetroses, heptoses and octoses), a disaccharide (e.g. sucrose, maltose, lactose, and trehalose) or an oligosaccharide or mixtures thereof with a fatty acid (s) with 4-22 carbon atoms (e.g. saturated fatty acids such as caprylic acid, capric acid, lauric acid myristic acid, p almitic acid and stearic acid, and saturated fatty acids such as palmitoleic acid, oleic acid, elaidic acid, erucic acid and linoleic acid)), or steroidal esters); alkyl, aryl or cyclic ethers with 2-30 carbon atoms (for example, diethyl ether, tetrahydrofuran, dimethyl isosorbite, diethylene glycol monoethyl ether); glycofurol (ether of polyethylene glycol and tetrahydrofurfuryl alcohol); ketones with 3-30 carbon atoms (e.g. acetone, methyl ethyl ketone, methyl isobutyl ketone); aliphatic, cycloaliphatic or aromatic hydrocarbons with 4-30 carbon atoms (e.g. benzene, cyclohexane, dichloromethane, dioxolanes, hexane, n-decane, n-dodecane, n-hexane, sulfolane, tetramethylene sulfone, tetramethylene sulfoxide, toluene, dimethyl sulfoxide) tetramethylene sulfoxide); alkyl or aryl halides containing 1-30 carbon atoms and optionally more than one halogen atom as a substituent; dichloromethane; monoethanolamine; petroleum ether; trolamine; omega-3 polyunsaturated fatty acids (e.g. alpha-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid or docosahexaenoic acid); polyglycol ether of 12-hydroxystearic acid and polyethylene glycol (Solutol® HS-15 from BASF, Ludwigshafen, Germany); polyoxyethylene glycerol; sodium laurate; sodium oleate; or sorbitan monooleate.

Preferably, as a pharmaceutically acceptable carrier in the present invention, proton-containing media such as water, saline solutions, water-soluble polymers, proteins, dextrose solutions (eg, DW5), electrolyte or alcohol solutions from the PAA Laboratory's catalog (2006) p. 26 are used. .

In a further aspect of the present invention, there is provided a kit for treating cancer, which comprises: (1) a pharmaceutical composition comprising a binuclear nitrosyl iron complex with the benzazheterocyclic derivatives of the present invention, in an airtight package; and (2) auxiliaries.

The kit may contain the composition in the form of a single dosage form or in the form of multiple doses. The kit may include forms for oral or parenteral administration.

The pharmaceutical composition in the kit can be placed in glass or polymer vials, ampoules, vials, metered dose cartridges for injectors, blisters, capsules, sachets with the composition used respectively for the oral or parenteral form.

Auxiliary agents include liquids for reconstituting a parenterally administered composition if it is provided in concentrated form in the kit, for example, in the form of a dry substance, a dried preparation, and the like; preparations for the preparation of oral liquid forms and injection forms ex tempore . Water for injection, physiological saline, lidocaine solution, etc., can be used as a recovery fluid. To restore the composition used in liquid form orally, a solution of glucose, sugars, syrups, etc. can be used.

Optional kit aids include openings for closures, means for sealing opened reusable closures, instruction inserts.

The pharmaceutical composition, which is a solid dosage form for oral administration, can be presented in a kit in the form of tablets, capsules in blisters, ampoules, vials, vials, sachets, and the like. The pharmaceutical composition, which is a liquid dosage form for parenteral or oral administration, may be presented in a kit in bottles, capsules, ampoules, cartridges, and the like.

An example of a kit for parenteral administration includes a package that contains instructions for use, ampoules or vials with a dry composition and ampoules with saline for injection. A device for opening ampoules is inserted into the package. Ampoules are packed in blisters of 10 ampoules.

Other kit options are obvious to a person skilled in the art from the above description.

The following examples are provided only as a further illustration of the invention and should not be construed as limiting the invention.

Examples of the synthesis of complexes

Example 1

Synthesis of complex (I), where R = benzimidazol-2-yl (Fe ₂ S ₂ N ₈ C _fourteen H ₁₀ O _four )

For the synthesis, the starting reagents Na ₂ S ₂ O ₃ · 5H ₂ O (Aldrich), NaOH (analytical grade), 2-mercaptobenzimidazole (Aldrich) were used. All operations for the preparation, mixing of solutions and the separation of complexes were carried out in an atmosphere of industrial argon at room temperature. An organic solvent was used for recrystallization.

A mixture of 0.992 g (4 mmol) of Na ₂ S ₂ O ₃ · 5H ₂ O and 1,074 g (2 mmol) of the thiosulfate nitrosyl complex of iron bis (μ-thiosulfato-S) -bis (dinitrosyl ferrate) (2-) sodium Na ₂ [Fe ₂ (S ₂ O ₃ ) ₂ (NO) ₄ ] (TNLC) was dissolved in distilled water. A mixture consisting of 1,500 g (10 mmol) of 2-mercaptobenzimidazole and 0.44 g (11 mmol) of NaOH was also dissolved in water.

Both solutions were mixed. The precipitate was filtered off and dried in air. The yield of intermediate was 0.719 g. After one day, the precipitate was recrystallized.

The complex is well soluble in ethanol, methanol, acetone, DMSO and DMF. Insoluble in dichloromethane, water, heptane and ether.

Yield: 63.50% ;

Found for Fe ₂ S ₂ N ₈ C ₁₄ H ₁₀ O ₄ ,%: Fe, 19.27; S, 10.69; N, 18.54; C, 43.40; H, 4.78; calculated,%: Fe, 18.99; S, 10.90; N, 19.05; C, 44.93; O, 16.32; H, 4.45.

IR spectrum (cm ^-1 ): 3207, 1787, 1738, 1515, 1467, 1428, 1384, 1272, 1218, 1183, 1002, 742, 713, 603.

Example 2

Synthesis of complex (II), where R = 5-methylbenzimidazol-2-yl (Fe ₂ S ₂ N ₈ C ₁₆ H _fourteen O _four )

For the synthesis, the starting reagents Na ₂ S ₂ O ₃ · 5H ₂ O (Aldrich), NaOH (bhd), 2-mercapto-5-methylbenzimidazole (Aldrich) were used. All operations for the preparation, mixing of solutions and the separation of complexes were carried out in an atmosphere of industrial argon at room temperature. An organic solvent was used for recrystallization. Distilled water was prepared for the synthesis by passing a stream of technical argon through it for 30 minutes.

A mixture of 0.4960 g (2 mmol) of Na ₂ S ₂ O ₃ · 5H ₂ O and 0.5740 g (1 mmol) of a thiosulfate nitrosyl iron complex of bis (μ-thiosulfato-S) -bis (dinitrosylferrate) (2-) sodium Na ₂ [Fe ₂ (S ₂ O ₃ ) ₂ (NO) ₄ ] (TNLC) was dissolved in distilled water. A mixture of 0.8250 g (5 mmol) of 2-mercapto-5-methylbenzimidazole and 0.2890 g (7 mmol) of NaOH was also dissolved in water.

Both solutions were mixed. The precipitate was filtered off and dried in air. The yield of intermediate was 2.2017 g. After one day, the precipitate was recrystallized.

The complex is well soluble in acetone, DMSO and DMF. Insoluble in dichloromethane, water, heptane and ether.

Yield: 45.53%;

Found for Fe ₂ S ₂ N ₈ C ₁₆ H ₁₄ O ₄ ,%: Fe, 19.27; S, 10.69; N, 19.54; C, 34.26; H, 2.48; calculated,%: Fe, 20.01; S, 11.49; N, 20.08; C, 34.43; O, 11.47; H, 2.53.

IR spectrum (cm ^-1 ): 3133, 1790, 1724, 1619, 1490, 1443, 1411, 1373, 1276, 1223, 1189, 1011, 854, 800, 720, 596, 543.

Example 3

Synthesis of complex (III), where R = benzthiazol-2-yl (Fe ₂ S _four N ₆ C _fourteen H ₈ O _four )

For the synthesis, the starting reagents Na ₂ S ₂ O ₃ · 5H ₂ O (Aldrich), NaOH (analytical grade), 2-mercaptobenzthiazole (Aldrich) were used. All operations for the preparation, mixing of solutions and the separation of complexes were carried out in an atmosphere of industrial argon at room temperature. An organic solvent was used for recrystallization. Distilled water was prepared for the synthesis by passing a stream of technical argon through it for 30 minutes.

A mixture of 0.4960 g (2 mmol) of Na ₂ S ₂ O ₃ · 5H ₂ O and 0.5740 g (1 mmol) of a thiosulfate nitrosyl iron complex of bis (μ-thiosulfato-S) -bis (dinitrosylferrate) (2-) sodium Na ₂ [Fe ₂ (S ₂ O ₃ ) ₂ (NO) ₄ ] (TNLC) was dissolved in distilled water. A mixture of 0.8402 g (5 mmol) of 2-mercaptobenzthiazole and 0.2890 g (7 mmol) of NaOH was also dissolved in water.

Both solutions were mixed. The precipitate was filtered off and dried in air. The yield of the intermediate was 1.1182 g. After a day, the precipitate was recrystallized.

The complex is well soluble in ethanol, methanol, acetone, methylene chloride, acetonitrile, ether, THF, DMSO and DMF. Not soluble in water, heptane.

Found for Fe ₂ S ₄ N ₆ C ₁₄ H ₈ O ₄ ,%: Fe, 18.96; S, 22.01; N, 14.54; C, 29.39; H, 1.57; calculated,%: Fe, 19.78; S, 22.72; N, 14.89; C, 29.79; O, 11.34; H, 1.43.

IR spectrum (cm ^-1 ): 3448, 2923, 2852, 2379, 1787, 1721, 1468, 1429, 1384, 1313, 1270, 1006, 852, 754, 724, 706, 611.

IR spectra of all samples were recorded on a SPECTRUM BX-II Fourier spectrometer. A sample was prepared in the form of tablets with KBr (1 mg of the analyte per 300 mg of KBr).

X-ray diffraction analysis of the Fe ₂ S ₂ N ₈ C ₁₄ H ₁₀ O ₄ (I) complex was carried out on a BRUKER P-4 automatic four-circle diffractometer (graphite monochromator, λ (Mo-K _α ) = 0.71073 Å, temperature 200 K, θ / 2θ scan). A black single crystal in the form of a parallelepiped measuring 0.35 × 0.20 × 0.15 mm was used in the experiment. The crystals are unstable at room temperature and are destroyed at a rate of 10% in 1 hour. The structure is deciphered by the direct method. The positions and thermal parameters of non-hydrogen atoms are refined in the isotropic and then anisotropic approximation by the full-matrix least-squares (LSM) method. Hydrogen atoms were detected from spaced Fourier syntheses and refined in the isotropic approximation.

The molecular structure of complex I is presented in figure 1. The complex has a centrosymmetric dimeric binuclear structure. In the dimer, two tetrahedrally coordinated by two NO groups and two benzazheterocyclic thiols, iron atoms are connected by a bridge: Fe-S-C-N-Fe΄. The complex contains two solvent molecules - acetone.

Crystallographic data and the main refinement parameters are presented in table 1.

Interatomic distances and angles are presented in table 2.

All calculations were performed using the SHELXTL software package (G.M. Sheldrick, SHELXTL v.6.14, Structure Determination Software Suite, Bruker AXS, Madison, Wisconsin, USA, 2000).

The distance Fe (1) ... ..Fe (1a) is 4,057 Ǻ cm (table 2).

Table 1
Crystallographic data and analysis characteristics for complex I Gross formula C ₂₀ H ₂₂ Fe ₂ N ₈ O ₆ S ₂ Molecular mass 646.28 Temperature 200 (2) K Wavelength 0,71073 Ǻ Syngonia, space group triclinic, P (-1) a 8,737 (1) Ǻ b 9,072 (1) Ǻ c 9,083 (1) Ǻ alpha 74.82 (1) ° beta 73.10 (1) ° gamma 86.62 (1) ° V 664.70 (13) Ǻ ³ Z, d _calc . one; 1.615 g / cm ³ μ 13.00 cm ^-1 F (000) 330 Crystal size 0.35 × 0.2 × 0.15 mm Θ shooting area 2.33-25.00 ° Total number of reflexes / independent 2572/2080 [R (int) = 0.0205] N / number of parameters 2080/216 GOOF by F ^ 2 0,994 R factor for reflections with I> 2 (σ) (I) R1 = 0.0270, wR2 = 0.0705 R factor for all reflections R1 = 0.0312, wR2 = 0.0731

table 2
Interatomic distances and valence angles in the structure of complex I Communication d, Ǻ Communication d, Ǻ Fe (1) -N (1) 1,682 (2) N (2) -O (2) 1,172 (3) Fe (1) -N (2) 1,687 (2) Fe (1) -N (3) # 1 2.0215 (18) O (1) -N (1) 1,165 (3) Fe (1) -S (3) 2,3029 (7) S (3) -C (1) 1,736 (2) Angle Ω, deg. Angle Ω, deg. O (2) -N (2) -Fe (1) 161.6 (2) N (2) -Fe (1) -S (3) 107.52 (7) O (1) -N (1) -Fe (1) 164.7 (2) N (3) # 1-Fe (1) -S (3) 109.73 (6) N (1) -Fe (1) -N (2) 111.05 (10) C (1) -S (3) -Fe (1) 102.48 (7) N (1) -Fe (1) -N (3) # 1 117.32 (9) C (1) -N (3) -Fe (1) # 1 126.14 (14) N (2) -Fe (1) -N (3) # 1 106.94 (9) C (3) -N (3) -Fe (1) # 1 127.76 (15) N (1) -Fe (1) -S (3) 103.92 (7) Communication d, Ǻ Communication d, Ǻ N (3) -C (1) 1,338 (3) C (7) -C (6)  1.378 (4) N (3) -C (3) 1,409 (3) C (3) -C (4)  1,387 (4) C (1) -N (4) 1.354 (3) C (4) -C (5)  1,394 (3) N (4) -C (2) 1,375 (3) C (2) -C (3)  1,401 (3) C (2) -C (7) 1,387 (3) C (6) -C (5)  1.389 (4) C (8) -C (10) 1,488 (4) C (8) -C (9)  1,492 (4) O (3) -C (8) 1,220 (3) Angle Ω, deg. Angle Ω, deg. C (1) -N (3) -C (3) 105.98 (17) C (4) -C (3) -C (2)  120.2 (2) N (3) -C (1) -N (4) 111.1 (2) C (4) -C (3) -N (3)  131.5 (2) N (3) -C (1) -S (3) 126.47 (16) C (2) -C (3) -N (3)  108.3 (2) N (4) -C (1) -S (3) 122.39 (19) C (3) -C (4) -C (5)  117.4 (2) C (1) -N (4) -C (2) 108.7 (2) O (3) -C (8) -C (10)  121.3 (2) N (4) -C (2) -C (7) 132.1 (2) O (3) -C (8) -C (9)  121.4 (2) N (4) -C (2) -C (3) 105.84 (19) C (10) -C (8) -C (9)  117.2 (3) C (7) -C (2) -C (3) 122.0 (2) C (7) -C (6) -C (5)  120.8 (2) C (6) -C (7) -C (2) 117.6 (2) Communication d, Ǻ Communication d, Ǻ C (4) -H (4) 0.96 (3) C (9) -H (9A) 0.96 (4) N (4) -H (1) 0.82 (3) C (9) -H (9B) 0.89 (4) C (6) -H (6) 0.95 (3) C (9) -H (9C) 0.90 (4) C (7) -H (7) 0.94 (3) C (10) -H (10A) 0.87 (6) C (5) -H (5) 0.88 (3) C (10) -H (10B) 0.99 (5) C (10) -H (10C) 0.96 (4) Angle Ω, deg. Angle Ω, deg. C (1) -N (4) -H (1) 124 (2) C (6) -C (5) -H (5) 120.4 (17) C (2) -N (4) -H (1) 127 (2) C (4) -C (5) -H (5) 117.6 (17) C (6) -C (7) -H (7) 126.8 (18) C (8) -C (9) -H (9A) 110 (2) C (2) -C (7) -H (7) 115.5 (18) C (8) -C (9) -H (9B) 109 (2) C (3) -C (4) -H (4) 122.7 (16) C (8) -C (9) -H (9C) 109 (2) C (5) -C (4) -H (4) 119.9 (16) C (8) -C (10) -H (10A) 114 (3) C (7) -C (6) -H (6) 121 (2) C (8) -C (10) -H (10B) 111 (3) C (5) -C (6) -H (6) 118 (2) C (8) -C (10) -H (10C) 111 (2) C (6) -C (5) -C (4) 122.0 (3)

Matrices of generation of equivalent atomic positions:

# 1 -x, -y, -z

The crystal structure of complex I is shown in FIG. 2.

The Mössbauer absorption spectra were recorded on a WissEl apparatus operating in a constant acceleration mode. The source was Co ⁵⁷ in the Rh matrix. Spectra were measured at low temperatures using a controlled temperature helium cryostat CF-506 (Oxford Instruments). The Mössbauer spectra were processed using the least squares method under the assumption of the Lorentzian shape of individual spectral components.

The results are presented in table 3.

Table 3
The NGR parameters of the spectra of Fe ⁵⁷ complexes I-III at T = 290 K Complex δ _Fe , mm / s ^-1 ΔE _Q , mm / s ^-1 I 0.202 0.954 II 0.232 0.805 III 0.217 0,994

Investigation of the NO-donor activity of binuclear nitrosyl iron complexes with benzazheterocyclic derivatives

The NO-donor activity of complexes I-III was first established by studying their reactions with hemoglobin (Hb). The research method is based on the reaction of the formation of Hb-NO in the interaction of hemoglobin with free NO in solution [R. Cassoly, QH Gibson, Conformation, co-operativity and ligand binding human hemoglobin, J. Mol. Biol. 91 (1975) 301-313; TJ McMahon, JS Stamler, Concerted nitric oxide / oxygen delivery by hemoglobin, in: L. Packer (Ed.), Methods In Enzymology, Academic Press, 301 (1999) Part C, 99-114]. The binding constant of Hb with NO is 3 · 10 ¹⁰ M ^-1 , which is three orders of magnitude higher than the corresponding constant for CO, and six orders of magnitude higher than the binding constant for O ₂ [E. Antonini, M. Brunori, Hemoglobin and myoglobin in the reactions with ligands, in: A. Neuberger, EL Tatum (Eds.) North-Holland research monographs. Fronties of biology. Vol. 21, North-Holland Publishing company, Amsterdam-London, 21 (1971) 276]. The bimolecular constant of interaction of Hb with NO is close to diffusion: 1.02 · 10 ⁸ M ^-1 · s ^-1 in 0.05 M phosphate buffer, pH7.0 at 20 ° С. Therefore, Hb is a very convenient trap for NO and is actively used to study the NO-donor activity of compounds generating nitrogen monoxide. In this case, the optical absorption spectra of free Hb and Hb-NO are very different, which makes it easy to detect the formation of a nitrosyl adduct.

The results of the study are presented for illustration purposes in figure 3 for complex I.

A homogeneous bovine Hb preparation was obtained from bovine hemoglobin from MP Biomedicals, a mixture of metHb and HbO ₂ . At all stages of Hb production and in all experiments with Hb, 0.05 M phosphate buffer, pH 7.0, was used. To convert a mixture of metHb with HbO ₂ into Hb, a 2 × 15 cm column with Sephadex G-25 was preliminarily prepared and transferred to an anaerobic state. At the outlet of the column, 5 ml of Hb with a concentration of 6 · 10 ^-4 mol · l ^-1 were collected. Hb was stored frozen in the form of balls in liquid nitrogen. Before use, Hb was thawed in 5 ml vessels in a stream of nitrogen. 2.8 ml of buffer, 0.1 ml of Hb 6 · 10 ^-4 mol · l ^-1 were introduced into the 4 ml anaerobic experimental cuvette with an optical path length of 1 cm and the absorption spectrum was recorded. Anaerobic absolute DMSO was added to the sample of the nitrosyl complex in the vessel filled with nitrogen, so as to obtain a solution of the complex with a concentration of 6 · 10 ^-3 mol · l ^-1 . Next, the solution was stirred for 3-5 minutes until completely dissolved and 0.1 ml of the resulting solution was introduced into an experimental Hb cuvette and into a comparison cuvette containing 2.9 ml of anaerobic buffer. The final concentration of the nitrosyl complex was equal to 2 · 10 ^-4 mol · l ^-1 , DMSO - 3.3%. Next, difference absorption spectra were recorded, the first one after 1 minute from the start of the reaction (shown in FIG. 3), then at intervals of 3 minutes during the first 30 minutes of the reaction, and then at intervals of 15, 30, 60 minutes (in FIG. 3 only part of these spectra is shown for clarity). Spectra were recorded until complete conversion of Hb to HbNO, when the spectrum ceased to change. The amount of NO formed was estimated spectrophotometrically by the amount of HbNO formed. To determine the concentration of HbNO, the absorption spectrum of the reaction system containing Hb and HbNO was decomposed by computer processing using the MathCad program into components — the spectrum of Hb and HbNO. The calculation was carried out in the wavelength range of 450-650 nm at 200 experimental points. To record the absorption spectra, a Specord M-40 spectrophotometer was used, equipped with an interface for computer recording of spectra and a thermostatically controlled cell compartment. Spectra were recorded at 25 ° C.

As a result of these studies, it was first established that the complexes generate NO in aqueous solutions of dimethisulfoxide spontaneously (i.e., in the absence of chemo, photo, or enzymatic activation).

In vitro study of cytotoxicity of tetranitrosyl iron complexes with benzazheterocyclic thiolyls on human tumor cells

Experimental technique

Before the experiment, all compounds were dissolved in 200 μl of DMSO, and then brought to the desired concentration with RPMI1640 nutrient medium. The final concentration of DMSO in the samples did not exceed 0.2% and did not affect cell growth.

Cell line - human ovarian cancer SKOV3. Cells were grown in a monolayer in RPMI1640 medium containing 10% fetal calf serum at 37 ° C and 5% CO ₂ content. For experiments, cells were seeded in 96-well plates and grown under the same conditions.

The cytotoxic effect was tested using the MTT test based on the ability of living cell dehydrogenase to restore the unpainted tetrazolium salt into blue formazan crystals soluble in dimethyl sulfoxide (DMSO).

All compounds were added to the wells in a volume of 20 μl at 4-final concentrations (μM) of 100, 50, 25, and 10. The total incubation volume was 200 μl. Cells with the preparations were incubated under the above conditions for 72 hours. At the end of the incubation, MTT reagent was added to the cells and incubated under the same conditions for 2 hours. Then, the formed formazan crystals were dissolved in 100 μl of DMSO at 37 ° C for 20 minutes. The optical absorption of DMSO solutions was measured on an optical counter for multi-well plates at a wavelength of 540 nm. The results were expressed as mean values for 4-parallel measurements. The cytotoxic effect was evaluated by cell survival in experimental samples relative to the control in%. A compound was considered active if, at a concentration of 100 μM, the number of living cells was 50% or less (IR ₅₀ ≤ 100 μM). The measurement error did not exceed 5%.

The results of the study are presented in table 4.

Two of the studied nitrosyl iron complexes, I and III, containing thiobenzimidazole and thiobenzothiazole ligands, showed cytotoxic activity on the SCOV ₃ cell line. For I and III, the IC ₅₀ was 57 μM and 25 μM, respectively.

The studied nitrosyl iron complexes I and III demonstrated a different cytotoxic effect on human tumor cells in vitro . The maximum activity was shown by complex III (IR ₅₀ = 25 μM).

As a reference drug, cisplatin (cis-DDP) was used, an antitumor drug used in clinics in Russia and abroad (IC ₅₀ = 16 μM). It was found that when the concentration is halved, complex III retains the same activity as at a concentration of 100 μM (tumor cell survival 12.5% (table 4). The fact that the toxicity of complex III is much lower (its total toxicity index LD ₁₀₀ is 50 mg / kg) than cisplatin (LD 16 = ₁₀₀ mg / kg), may have a value in the high-dose therapy accompanied by serious side effects, as a way of increasing the sensitivity of tumor cells to therapeutic effects, and will reduce the concentration Applicability cytostatics.

Table 4
Cytotoxic activity of nitrosyl iron complexes I-III on SKOV3 cell line No. p / p Concentration, μM Compound. Cell survival,% I II III one one hundred 12,4 one hundred 12.5 2 fifty 56.32 one hundred 12.5 3 25 87.5 one hundred 50.1 four 10 one hundred one hundred 88.5 5 IR ₅₀ , μm 57 > 100 25

In accordance with the guidelines for studying the antitumor activity of pharmacological substances, studies were conducted on transplantable tumors of mice on Ca-755, lymphocytic leukemia P-388 and melanoma B-16 (on first-generation hybrid mice BDF ₁ (C ₅₇ Bl / 6 × DBA / ₂ ) and DBA / ₂ weighing 18-25 g, obtained from the department of laboratory animals of the GU RONTs im.NNN Blokhin RAMS). A statistically significant antitumor effect of the claimed drugs was revealed.

Claims (16)

1. Binuclear nitrosyl complex of iron with benzazheterocyclic derivatives of the General formula [Fe ₂ (SR) ₂ (NO) ₄ ],
where r

and where X is NH, S, R ¹ is lower alkyl. 2. The binuclear nitrosyl iron complex according to claim 1, where R is benzimidazol-2-yl, 5-methyl-benzimidazol-2-yl or benzthiazol-2-yl. 3. The method of producing binuclear nitrosyl iron complexes according to claim 1, wherein the thiosulfate nitrosyl iron complex is treated with the corresponding benzazheterocyclic thiol in a stoichiometric ratio in the presence of a reducing agent, and the process is carried out in an alkaline medium, followed by isolation of the target product by known methods. 4. The method according to claim 3, characterized in that the process is carried out at room temperature, mainly at 18-25 ° C. 5. The method according to claim 3, characterized in that the process is conducted in an oxygen-free atmosphere. 6. The method according to claim 3, characterized in that benzimidazole-2-thiol, 5-methyl-benzimidazole-2-thiol or benzthiazole-2-thiol are used as the benzazheterocyclic thiol. 7. The method according to claim 3, characterized in that hydrogen, metal thiosulfate, hydrogen sulfide and aliphatic thiols are used as reducing agent. 8. The donor of nitrogen monoxide, which is a binuclear nitrosyl iron complex according to claim 1. 9. The use of binuclear nitrosyl iron complex according to claim 1 as an antitumor drug. 10. The use of binuclear nitrosyl iron complexes according to claim 1 for obtaining an antitumor drug. 11. A pharmaceutical composition comprising an effective amount of a binuclear nitrosyl iron complex with a benzazheterocyclic derivative according to claim 1 and a pharmaceutically acceptable carrier as an antitumor drug. 12. The pharmaceutical composition according to claim 11, where a proton-containing medium is used as a pharmaceutically acceptable carrier. 13. The pharmaceutical composition according to claim 11, where as a pharmaceutically acceptable carrier using a mixture of proton-containing medium and dimethyl sulfoxide. 14. The pharmaceutical composition according to any one of claims 12 or 13, wherein water, physiological saline, and water-soluble biopolymers are used as the proton containing medium. 15. The pharmaceutical composition according to claim 11, where the binuclear nitrosyl complex of iron with benzazheterocyclic derivatives is present in an amount of 50-100 μm. 16. A kit used to treat cancer, including: (1) a pharmaceutical composition comprising a binuclear nitrosyl iron complex with benzazheterocyclic derivatives according to claim 1, in an airtight package; and (2) auxiliary components.

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