Water-soluble binuclear cation nitrosyl iron complexes with natural aliphatic thiolyls possessing cytotoxic, apoptotic and no-donor activity

FIELD: pharmacology.

SUBSTANCE: invention refers to binuclear cation nitrosyl iron complexes with natural aliphatic thiolyls with general formula [Fe2(SR)2(NO)4]SO4 where R is aliphatic ligands of natural origin.

Also there is proposed method for production of binuclear cation nitrosyl iron complex, nitrogen monoxide donor, inductor of tumor cells apoptosis, application binuclear cation nitrosyl iron complex, pharmaceutical composition and set used for treatment of oncology diseased. The technical result is EFFECT: production of binuclear cation nitrosyl iron complex possessing cytotoxic, apoptotic and NO-donor activity.

15 cl, 2 ex, 4 tbl, 7 dwg

 

The technical field

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

Prior art

In recent years, with the aim of creating drugs of new generation, used in the treatment of cancer, is an intensive search for anticancer agents based on transition metal complexes with improved spectrum of activity and reduced side effects in comparison with the already used clinical drugs, such as cisplatin, sodium nitroprusside, etc.

The use of NO donors as a new class of antitumor agents is associated with the important role of NO in the process of growth of malignant tumors [Wink D., Vodovoz J., Cook, J., Biochemistry, 1998, 63, 7, pp. 948-957]. It is shown that the nitric monoxide changes the level of apoptosis of tumor cells, the activity of a gene p53 and neoangiogenesis [Brune B., Scheneiderhan N., Nitric oxide evoked p53 accumulation and apoptosis, Toxicol Letters, 2003, 193, 2, RR-23], and inhibits the activity of key repair protein O6-methyl-guanine-DNA-methyl-transferase 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, R-3043]. However, the known time and synthetic NO-donors of different classes (organic nitrates, diatonically, nitrosothiol and others) are not used as clinical therapeutic agents for the treatment of cancer, and are used only for reinforcement (in varying degrees, depending on the chemical nature 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., H. Ding, 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].

On the other hand, in the last 15 years firmly established that one of the forms of natural reservoirs are NO nitrosyl iron complexes with sulfur-containing ligands. Such compounds are formed in the cells as a result of the actions of endogenous NO on the active centres are not heme iron-sulfur proteins [A.R. Butler, Megson I.I., Chem. Rev., 2002, 102, pp.1155-1165] and are the cell “depot” NO. Binding of NO to the active centers of metallothionen, in particular, with megamovie iron-containing proteins intensivelystudied[Ford P.C., Lorkovic I.M., Chem.Rev., 2002, 102, 993; Hoshino, M.; Laverman LE, Ford P.C., Coord. Chem. Rev., 1999, 187, R].Their synthetic models - ethers “red salt” Russa have the composition [Fe2(SR)2(NO)4], where R=Et, t-Bu, (CH2)4-CH3C 6H5F, Ph [T. Thomas, J.H. Robertson, E.G. Cox, Acta. Crystalogr., 1958, 11, p.599; C. Glidewell, M.E. Harman, M.B. Hursthouse, I.L. Johnson, M. Motevalli, J. Chem.Res., 1998, 212, p.1676; R.E. Marsh, A.L. 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; T. B. Rauchfuss, T.D. Weatherill, Inorg. Chem., 1982, 21,pp. 827-830.]. These binuclear diamagnetic sulfur-nitrosyl complexes µ-S type R=Alk generate NO in thermo - or photoactivation [J.L. Bourassa, P.C. Ford, Coord. Chem.Rev., 2000, 200-202, p.p. 887-900] and can serve as a new promising anticancer NO-generousity agents. In the work of A. Janczyk, et.all. Nitric Oxide, 2004, 10, 1, pp. 42-50 first studied by direct cytotoxic action of nitrosyl iron complex Na[Fe4S3(NO)7] on human melanoma cells in human and mouse. However, the nitrosyl complex of iron generates NO only when photoactivation, and cannot be used as anticancer drugs because of the high toxicity against normal cells.

Also known binuclear paramagnetic sulfur-nitrosyl iron complexes with aromatic ligands µ-N-C-S type [N.A. Sanin, S.M. Aldoshin ”Functional model nitrosyl [Fe-S] proteins”// Invensure. (2004) 11 2326-2345; N.A. Sanin, S.M. Aldoshin, TN. Rudnev, N. Golovin, Shilov G.V., Y.M. Shulga, V. M. Martynenko, NS Hovhannisyan. ”Synthesis, structure and solid-phase transformations of nitrosyl iron complex a 2[Fe2(S2O3)2(NO)4]∙4H2O”, Coordination Chemistry, 2005, 31, 301-306; N.A. Sanina, TN Rudneva, S.M. Aldoshin, Shilov G.V., DV Korchagin, Yu.M. Shul'ga, V.M. Martinenko, N.S. Ovanesyan, “Influence of CH3 group in 1-methyl-imidazole-2-yl on the properties of binuclear sulfure-nitrosyl iron complex with the ligand of µ-N-C-S type”//Inorganica Chimica Acta (2006), 359, 2, 570-576; N.A. Sanin, TN. Rudnev, S. M. Aldoshin, A. N. Covers, R. B. Morgunov, E. C. Kurganov, N. S. Hovhannisyan, “Synthesis, structure, and NO-donor activity of the paramagnetic complex [Fe2(SC3H5N2)2(NO)4] as model nitrosyl [2Fe-2S] proteins”//proceedings of the Academy of Sciences. The series of chemical,1(2007) 28-34; 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], which spontaneously generate NO in proton environments and, as a recent study has shown, can be used as vysokoeffektivnye donors NO chemotherapy [N.A. Sanin, O.S Zhukov, SS Smirnova, T.N. Rudnev, Shilov G.V., S.M. Aldoshin “Binuclear nitrosyl iron complexes with besuseradminclient derivatives, method of receipt, the donor of nitrogen monoxide, pharmaceutical composition containing them and their use as anticancer drugs - application number PCT/RU2007/000286 dated may 30, 2007;N.A. Sanin, O.S Zhukov, S.M. Aldoshin, NS Yemelianova, G. Gerasimov, “Application tetranitroaniline complex of iron with tiofenolom as anticancer drugs and pharmaceutical compositions and kits, containing this complex application number PCT/RU2007000285 from 30 may 2007]. However, their widespread use in the clinic is limited by their low solubility in water and physiological fluids. Thus, there is a need for water-soluble (and thus more bioavailable) antitumor tools with increased efficacy and reduced toxicity. In [S.A.T. Dillinger, H.W.Schmalle, T.Fox, H.Berke, “Developing iron nitrosyl complexes as NO donor prodrugs”//Dalton Trans., (2007)3562-3571] described the synthesis of binuclear cationic nitrosyl iron complexes with group probably facilitates composition [Fe(NO)2S(CH2)2NH3]2X2(X=Cl, I): to learn the basic products needed to conduct the reaction in a toxic tetrahydrofuran using multistage received dinitrosyl iron - Fe(CO)2(NO)2or [Fe(NO)2I]2.

The present invention is to expand the Arsenal of anticancer agents and the creation of anticancer drugs based on nitrosyl iron complexes with improved spectrum of activity and reduced side effects, in particular water-soluble binuclear nitrosyl iron complexes with sulfur-containing aliphatic ligands of natural origin, preferably the group probably facilitates penicillamine), applicable as NO-donors and with increased activity and reduced toxic the flesh.

A brief description of the nature of the invention

In one aspect the invention relates to new water-soluble binuclear nitrosyl cation of the iron complexes of the formula [Fe2(SR)2(NO)4]SO4,

where R represents a sulfur-containing aliphatic ligands of natural origin, such as the group probably facilitates penicillamine, cysteine, S-sulfenylation, various cysteinsulfinic, N-acetyl-DL-penicillamine, N-acetylcysteine, thioglycolate acid, mercaptosuccinic acid, methionine, methanesulfonic, methanesulfonic, S-adenosylmethionine, glutathione and other Preferably R represents a group probably facilitates or penicillamine.

In another aspect the invention relates to a method for producing water-soluble binuclear cationic nitrosyl iron complexes with sulfur-containing aliphatic ligands of natural origin formula [Fe2(SR)2(NO)4]SO4treatment of an aqueous solution of iron sulfate (II) the corresponding water-soluble tiamina in the stoichiometric ratio in the presence of gaseous NO in an acidic environment. 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 monoxide nitrogen, which represents a binuclear nitrosyl complex glands is water-soluble sulfur-containing aliphatic ligands, above.

The following aspect of the invention relates to an inductor of apoptosis of tumor cells, which represents a binuclear nitrosyl complex of iron with water-soluble sulfur-containing aliphatic ligands as described above.

In the following aspect the present invention concerns the use of binuclear nitrosyl iron complexes with sulfur-containing aliphatic ligands of natural origin formula [Fe2(SR)2(NO)4]SO4as anticancer drugs.

In an additional aspect, the present invention relates to the use of binuclear nitrosyl iron complexes with sulfur-containing aliphatic ligands of natural origin formula [Fe2(SR)2(NO)4]SO4to obtain anticancer drugs.

In the following aspect the present invention relates to pharmaceutical compositions containing an effective amount binuclear nitrosyl iron complex with sulfur-containing aliphatic ligands of natural origin formula [Fe2(SR)2(NO)4]SO4and pharmaceutically acceptable carrier.

In the following aspect the present invention relates to a kit used for the treatment of cancer, containing (1) farm is tsevtichesky composition, including binuclear nitrosyl complex of iron with sulfur-containing aliphatic ligands of natural origin formula [Fe2(SR)2(NO)4]SO4where R has the above meanings, in a sealed package and (2) auxiliary tools.

Description of figures

Figure 1 shows the molecular structure of the complex [Fe2(S(CH2)2NH3)2(NO)4]SO4·2,5H2O (complexI).

Figure 2 presents the crystal structure of the complexI.

Figure 3 presents the molecular structure of complex [Fe2(S(C(CH3)2CH(NH3)COOH))2(NO)4]SO4·5H2O (complexII).

4 shows the crystal structure of the complexII.

Figure 5, 6 shows the dependence of the concentration of NO from time to time by the decomposition of the complexesIandII(4·10-6M) in water under anaerobic conditions at pH 6.25 and 25°C.

Figure 7 presents the analysis of the fluorescent signal inI-50 induced cells: A graphic image in the coordinates of the forward (FSC) and transverse (SSC) light scattering induced by the uncolored cell line K-562 (control); B - induced autofluorescence of unstained cells; induced fluorescence stained cells.

Detailed description the s inventions

New binuclear cationic nitrosyl iron complexes with water-soluble Mililani, according to the present invention have the General formula [Fe2(SR)2(NO)4]SO4where R represents a sulfur-containing aliphatic ligands of natural origin, such as the group probably facilitates penicillamine, cysteine, S-sulfenylation, various cysteinsulfinic, N-acetyl-DL-penicillamine, N-acetylcysteine, thioglycolate acid, mercaptosuccinic acid, methionine, methanesulfonic, methanesulfonic, S-adenosylmethionine, glutathione and other

Preferably R represents a group probably facilitates or penicillamine.

In the prior art there is no information about the claimed compounds and the way they are received.

Way to generate new binuclear nitrosyl iron complexes with sulfur-containing aliphatic ligands of natural origin formula [Fe2(SR)2(NO)4]SO4where R has the above significance, is that an aqueous solution of iron sulfate (II) is treated with an appropriate water-soluble tiamina in the stoichiometric ratio in the presence of gaseous NO and the process is conducted in an acidic medium followed by separation of the target product by known methods.

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

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

As water-soluble thiamine preferably used aliphatic amines of natural origin - group probably facilitates penicillamine.

The inventors have found that the binuclear nitrosyl iron complexes with sulfur-containing ligands of natural origin present invention are effective NO donors, generating nitric monoxide spontaneous decomposition in proton environments (such as water, blood and its components, physiological solutions, and the like), in the absence of chemo-, photo - or enzyme activation. Thus, the complexes of the present invention represent a promising new class of donors of nitrogen monoxide - synthetic analogues of the active sites nitrosyl Nehemiah iron-sulphur proteins - natural “depot” NO. Therefore, the present invention in an additional aspect relates to the donors of nitrogen monoxide, which represents a binuclear nitrosyl iron complexes with water-soluble ligands of natural origin, presented above. Binuclear nitrosyl iron complexes with ligands of natural origin present invention have antitumor activity against human tumor cells.

The inventors have also found that the bi is gender nitrosyl iron complexes with ligands of natural origin present invention have apoptotic activity against tumor cells. Therefore, the present invention in an additional aspect relates to inducers of apoptosis of tumor cells, which represents a binuclear nitrosyl iron complexes with water-soluble ligands of natural origin, above.

The present invention is further directed to the use of binuclear nitrosyl iron complexes of the formula [Fe2(SR)2(NO)4]SO4where R has the above significance, as anticancer drugs. In particular, the compounds of the present invention can be used to treat erythroblastosis human leukemia of K 562 line cells, ovarian cancer, mammary adenocarcinoma (CA -755, leukemia P-388, melanoma B-16, epidermoid carcinoma Lewis. The iron complexes according to the present invention are suitable for inhibiting the growth of tumors in mammals, and is preferably introduced in the form of a pharmaceutical composition comprising an effective antitumor amount of a compound according to the present invention in combination with at least one pharmaceutically or pharmacologically acceptable carrier and/or excipient. The media is also known from the prior art as excipient, filler, excipient, additive or diluent, is any substance, which is supplied with the ski inert, gives appropriate consistency or form to the composition, and does not reduce therapeutic efficacy of antitumor compounds. The media is a "pharmaceutically or pharmacologically acceptable"if it does not cause any adverse, allergic or other adverse reactions when administered to a mammal or human, respectively.

The present invention additionally provides pharmaceutical compositions containing an effective amount binuclear nitrosyl iron complex with water-soluble ligands of natural origin formula [Fe2(SR)2(NO)4]SO4where R has the above significance, and a pharmaceutically acceptable carrier.

Preferably as a pharmaceutically acceptable carrier in a composition according to the invention is used protonotariou environment.

Preferably, as protonotaria environment using water, saline solution, water-soluble biopolymers.

Preferably, binuclear nitrosyl complex of iron with water-soluble ligands of natural origin present in the composition in amount of 50-100 microns.

Pharmaceutical compositions containing protivoopuxolevye compounds according to the present invention can be obtained by any conventional method. Necessary preparative form in biretta depending on the selected method of administration. Compositions according to the invention can be prepared for any way of introduction so that the target tissue was available in 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 intrasternally), local (nasal, percutaneous, ocular), vnutrishkolnoe, vnutriobolochechnoe, small intestine, lung, intralymphatically, intracavitary, vaginal, transurethrally, intradermal, ear, intramammary, transbukkalno, orthotopic, intratracheal, intranidus, percutaneous, endoscopic, cressilia, sublingual and intestinal introduction.

Pharmaceutically acceptable carriers for use in the compositions according to the present invention are well known to the specialist, and they are chosen depending on a number of factors: specific antitumor compound and its concentration, stability and expected bioavailability; the disease, disorder or condition of the person being treated with the composition; the subject, its age, weight and General condition; and the route of administration. Suitable carriers can be easily determined by the expert [J.G. Nairn in: Remington's Pharaceutical Science (ed. A. Gennaro), Mack Publishing Co., Easton, Pa, 1985, RR 1492-1517].

The composition preferably receive in the form of tablets, dispergiruyushchie powders, pills, capsules, gelatin capsules, dripping with the floor, gels, liposomes, granules, solutions, suspensions, emulsions, syrups, elixirs, tablets, pills, cakes or any other dosage forms that can be administered orally. Compositions according to the present invention for oral administration include an effective antitumor amount of a compound according to 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, carboxymethylcellulose, agar, lures, sorbitol, calcium phosphate, calcium carbonate, sodium carbonate, kaolin, alginic acid, gum Arabic, corn starch, potato starch, sodium saccharinate, magnesium carbonate, tragakant, microcrystalline cellulose, colloidal silicon dioxide, nitrocresols, talc, magnesium stearate and stearic acid. Further, such solid dosage forms may be uncoated or can be coated with a known coating methods, for example, to delay disintegration and absorption. Antitumor compounds according to the present invention, also preferred is a recreational use for the preparation of dosage forms for parenteral administration, for example, in the form of dosage forms for injection intravenous, intraarterial, subcutaneous, rectal, intramuscular, intraorbital, intracapsular, intraspinal, intraperitoneal or intrasternally ways. Compositions according to the invention for parenteral administration include an effective antitumor amount of antitumor compounds in a pharmaceutically acceptable carrier. Suitable for parenteral dosage forms include solutions, suspensions, dispersions, emulsions, or any other dosage form, which may be introduced parenterally. Methods and compositions for receiving parenteral dosage forms are known from the prior art.

In the composition according to the invention may be included for various purposes a small number of additional components that are well known in the pharmaceutical industry. These components are for the most part give properties that increase the retention time of antitumor compounds in the introduction, we contribute to the stability of the composition, regulate pH value, facilitate the introduction of antitumor compounds in pharmaceutical compositions, etc. Each of these components individually is present in quantities of preferably less than about 1 wt.%, more preferably less than about 5 wt.%, and most preferably less than about 0.5 wt.%, calculated on the total weight of the composition. Some components, such as fillers or diluents, can be up to 90 wt.%, calculated on the total weight of the composition, as is well known in the technology of preparation of medicines. Such additives include createsite components to prevent the deposition of iron complex, surfactants, wetting or emulsifying agents (e.g. lecithin, Polysorbate-80, tween®80, pluronic-60, polyoxyethylenated), preservatives (such as ethyl-p-hydroxybenzoate), protects from the effects of microbes means (as, for example, benzyl alcohol, phenol, m-cresol, chlorobutanol, sorbic acid, thimerosal, and paraben), agents for adjusting the pH or buferiruemoi agents (as, for example, acids, bases, acetic sodium, sorbitan-monolaurate), components for control osmolarity (as, for example, glycerin), thickeners (for example, aluminum monostearate, stearic acid, cetyl alcohol, stearyl alcohol, guar gum, methylcellulose, hydroxypropylcellulose, tristearin, complex citylove esters wax, polyethylene glycol), pigments, dyes, additives, giving fluidity, fixed specification of the HN ( for example, cyclomethicone), clays (such as bentonite), adhesives, increasing the volume of agents, flavorings, sweeteners, adsorbents, fillers (such as, for example, water, saline, solutions of electrolytes), binders (for example, water, saline, solutions of electrolytes), binders (such as, for example, starches such as corn starch, wheat starch, rice starch or potato starch, gelatin, tragakant, methylcellulose, hypromellose, sodium carboxymethyl cellulose, polyvinylpyrrolidone, sugars, polymers, gum Arabic), loosening agents (as, for example, starches such as corn starch, wheat starch, rice starch, potato starch or carboximetilkrahmal, structured polyvinylpyrrolidone, agar, alginic acid or its salt, such as sodium alginate, nitrocresols or crosspovidone), lubricating agents (such as, for example, silicon dioxide, talc, stearic acid or its salts, such as magnesium stearate, or polyethylene glycol), agents for coatings, such as concentrated sugar solutions, including gum Arabic, talc, polyvinylpyrrolidone, carbopol, polyethylene glycol, or titanium dioxide) and antioxidants (such as, for example, sodium metabisulfite, sodium bisulfite, sodium sulfite, dextrose, phenols and thiophene is ly).

Also, it is preferable that the antitumor compound mattered ID100(that is, the concentration of drug causing 100%inhibition of the formation of the colony), at least 4 times less than that of cisplatin, when carrying out measurements in accordance with the methodology described in ["Experimental evaluation of anticancer drugs in the USSR and the USA". /Ed. by Z.P. Sofianou, A.B. sarcina (USSR), A. Goldin, A. Klein (USA). M: "Medicine", 1979 p.71-105].

The introduction of the dosage forms listed ways can be continuous or intermittent, depending, for example, on the physiological state of the patient, whether the purpose of the introduction of therapeutic or prophylactic, and other factors, known or estimated practitioner.

Dose and scheme to introduce the pharmaceutical compositions according to the invention can be easily determined by the oncologist. It is clear that the dose of anticancer compounds depends on age, sex, health and weight of the recipient, type the concurrent treatment, if any, frequency of treatment and the nature of the desired effect. For any way of introduction, the exact amount used anticancer compounds, as well as appointed by the dose required to achieve the described here useful effects, also depends in particular on factors such as bioavailability of anticancer compounds, a human disease that is treated, the desired therapeutic dose, and other factors that are obvious to the expert.

The concentration of anticancer compounds in the liquid pharmaceutical composition is most preferably 50-100 μm. Generally, the preferred relatively low concentration, so as anticancer compound is most soluble at low concentrations.

Solutions for parenterale injection can be prepared by dissolving an anticancer compounds in water and other protonotaria environments (salt solutions, solutions of sugars, water-soluble polymers, proteins, electrolytes) for parenteral administration to a patient.

So, for example, preparative liquid forms as a carrier usually use water, salt solutions, solutions of dextrose (for example, DW5), solutions of electrolytes or any other proton, pharmaceutically acceptable environment.

Preferably as a pharmaceutically acceptable carrier in the present invention can be used protonotaria environment, such as water, saline solutions, water-soluble polymers, proteins, solutions of dextrose (for example, DW5), electrolyte solutions or alcohols from the catalogue “PAA Laboratory's” (2006) R.26.

In SL is blowing aspect of the present invention features a kit for the treatment of cancer, which includes (1) a pharmaceutical composition comprising a binuclear nitrosyl iron complex with sulfur-containing ligands of natural origin present invention, in a sealed package and (2) auxiliary tools.

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

Pharmaceutical composition in the kit can be placed in glass or plastic vials, ampoules, bottles, measured cartridges for injectors, blisters, capsules, sachets with the composition used, respectively, for oral or parenteral form.

Auxiliary means include fluid recovery composition introduced parenterally, if she's in the set is presented in a concentrated form, for example, in the form of solids, dried drug etc.; means for receiving oral liquid forms and forms for injectionex tempore. As a liquid for recovery can be used water for injection, saline solution, lidocaine and the like To restore the composition applied in liquid form by mouth, can be used glucose, sugars, syrups, etc. Optional auxiliary tool set including the means to indicate the opening of youporon, means for sealing the opened reusable youporon, liners statement.

Pharmaceutical composition, which represents a solid dosage form for oral administration may be presented as a set in the form of tablets, capsules in blisters, vials, bottles, vials, bags, etc. Pharmaceutical composition constituting the liquid dosage form for parenteral or oral administration may be presented as a set in vials, capsules, ampoules, cartridges, etc.

Example set for parenteral administration includes packaging, which includes instructions for use, ampoules or vials with dry composition, and vials with saline solution for injection. In the packaging goes a device for opening ampoules. The ampoules are Packed in blisters of 10 vials.

Other options set obvious to the person skilled in the art from the above description.

The following examples are provided only as an additional illustration of the invention and they should not be construed as limitations of the invention.

Examples of the synthesis of complexes

Example 1

Sulfate di-μ-contaminationpesticides (Fe2S3N6C4H19O10,5) (I)obtained by the reaction of dissolved in water is of Ulfat iron (II) c is a solution of the group probably facilitates hydrochloride. Distilled water used in the synthesis, requires advance preparation to remove dissolved oxygen. To do this, through the flask with distilled water is passed a current of inert gas (argon or nitrogen) for 30 minutes. 0.84 g (3 mmol) semimodule iron sulfate was dissolved in 12 ml prepared with distilled water. To this solution was added 0.40 g (3.5 mmol) of the group probably facilitates hydrochloride, dissolved in 8 ml of water. Through the resulting solution was passed a current of nitrogen monoxide obtained by the method [Y. Karjakin and I.I. Angels. Clean chemical substance. M.: Chemistry, 1974, 23] for 1-1,5 hours at ~40aboutWith prior education in the solution of the crystals. The reaction mixture is then kept at 6-8°C for 15-20 h, the formed fine crystalline precipitate a brown-red color was filtered and dried in the air during the day. The output was 0,143 mg (16.5 percent). The complex is well soluble in water and DMSO, worse in methanol, in other organic solvents (ethanol, THF, heptane, acetonitrile, ether and so on) is not soluble. The polycrystals of this complex is quite stable and stored in air at room temperature for quite a long time.

Found, %: Fe, 21,57; S, 18,39; N, 16,34; C, 9,40; H, 2,38.

Calculated, %: Fe=21,58; S=18,50; N=16,18; C=9,25, O=32,37; H=2,12.

IR-spectrum (in KBr pellets), (cm-1): 3454 (Ms.), 3003 (weak.), 2927 weak.), 1769 (sin.), 1727 (sin.), 1461 (weak.), 1385 (Ms.), 1340 (Ms.), 1266 (Ms.), 1120 (sin.), 770 (weak.), 620 (sin.).νNO: 1769 and 1727 cm-1.

Example 2

Sulfate di-μ-penicillinresistant (Fe2S3N6C10H32O17) (II)obtained by the reaction of iron sulfate (II) c D-penitsillaminom in the ratio of 1:3. The reaction was carried out using standard vacuum line and technology Slanka in an argon atmosphere at room temperature. From water previously removed the oxygen method three freezing and pumping in a vacuum. To the dry mixture, containing 0.42 g (1.5 mmol) semimodule iron sulfate and 0.68 g (4.5 mmol) of D-penitsillamin, was added 10 ml of the prepared water. Through the resulting solution dark purple color missed nitric oxide, obtained by the known method [Y. Karjakin and I.I. Angels. Clean chemical substance. M.: Chemistry, 1974, 23]. During the reaction the solution has gained a deep red color. After 10-12 minutes on the walls of the reaction vessel appeared fine needle crystals of red, gradually filling the entire volume of the solution. They were filtered under vacuum and dried under argon. The output was: 98 mg (11.3 per cent). The resulting product is stable in the absence of an inert atmosphere for a long time. The remaining mother liquor was kept e is e three days at a temperature of 6-8°C. Thus obtained red-orange needle crystals were suitable for x-ray analysis (PCA).

Found, %: Fe,15.62 Wide; S,13,34; N,11,89; C,Ls 16.80, H,4,62.

Calculated, %: Fe=15,64; S=13,42; N=11,72; C=16,76, O=37,99; H=4,47.

IR-spectrum (in KBr pellets), (cm-1): 1771 (sin.), 1723 (sin.), 1626 (Ms.), 1375 (Ms.), 1337 (Ms.), 1269 (Ms.), 1189 (Ms.), 1114 (Ms.), 1089 (Ms.), 746 (cf.). νNO: 1771 and 1723 cm-1.

X-ray diffraction analysis of complexes I and IIperformed on diffractometer SMART APEX2 CCD (Bruker) [λ(MoKα)=0,71072Å, ω-scans, 2θ<56°] at 100.

Crystallographic data and the main parameters of the refinement are given in tables 1, 2. Interatomic distances and angles are presented in tables 3, 4. Crystallochemical parameters depositiona in the Cambridge crystallographic given Bank (CCDC forI-663194 forII-680286).

Table 1
Crystallographic data and characteristics analysis for complexI
Brutto-formulaC4H19Fe2N6O10.50S3
Molecular weight527,13
Temperature100(2) K
Wavelength,71073 Å
The crystal system, space groupTriclinic, P -1
a6,8503(11) Å
b10,5747(16) Å
c13,723(2) Å
alpha90,884(3)°,
beta95,900(3)°,
gamma90,635(3)°,
V988,6(3) Å3
Z, dthe calc.2, 1,771 mg/m3
µ1,839 mm-1
F(000)538
The crystal size0,35×0,04×0.03 mm3
The image area for θfrom 1,93 up 28,00°
The total number of reflexes/independent5969
N/a number of arguments
GOOF on F^20,930
R factor for reflections with[I >2(is)(I) R1=0,0548, wR2=0,1058
R factor for all reflectionsR1=0,1053, wR2=0,1204

Table 2
Crystallographic data and characteristics analysis for complexII
Brutto-formulaC10H32Fe2N6O17S3
Molecular weight716,30
Temperature100(2) K
Wavelength0,71073 Å
The crystal system, space groupMonoclinic, P21
a6,1878(4) Å
b28,2739(17) Å
c7,8615(5) Å
alpha90°
beta102,4557(14)°
gamma90°
V1343,02(15) Å3
Z the calc.2, 1,771 mg/m3
µ1,397 mm-1
F(000)740
The crystal size0,36×0,10×0.04 mm3
The image area for θfrom 2.65 to 28,99°
The total number of reflexes/independent6900
N/a number of arguments
GOOF on F^21,029
R factor for reflections with[I>2(σ)(I)R1=0,0392, wR2=0,0811
R factor for all reflectionsR1=0,0491, wR2=0,0854

td align="justify"> 2,2444(15)
Table 3
Interatomic distances and valence angles in the structure of the complexI
CommunicationǺCommunicationǺ
Fe(1)-N(2)1,660(5)Fe(1)-N(1)1,675(5)
Fe(1)-S(1)Fe(1)-S(1)#12,2525(15)
Fe(1)-Fe(1)#12,6725(15)S(1)-C(1)1,837(5)
S(1)-Fe(1)#12,2525(15)O(1)-N(1)1,164(5)
O(2)-N(2)1,180(5)N(3)-C(2)1,490(6)
N(3)-H(3NA)1,0315N(3)-H(3NC)0,8500
N(3)-H(3NB)0,8500C(1)-C(2)1,504(7)
C(1)-H(1A)0,9900C(1)-H(1B)0,9900
C(2)-H(2A)0,9900C(2)-H(2B)0,9900
Fe(1')-N(1')1,667(4)Fe(1')-N(2')1,668(5)
Fe(1')-S(1')#22,2447(15)Fe(1')-S(1')2,2507(15)/td>
Fe(1')-Fe(1')#22,6823(15)S(1')-C(1')1,826(5)
S(1')-Fe(1')#22,2447(15)O(1')-N(1')1,175(5)
O(2')-N(2')1,172(5)N(3')-C(2')1,478(6)
N(3')-H(3NF)0,8500N(3')-H(3ND)0,8499
N(3')-H(3NE)0,9527C(1')-C(2')1,500(7)
C(1')-H(1'A)0,9900C(1')-H(1'B)0,9900
C(2')-H(2'A)0,9900C(2')-H(2'B)0,9900
S(1S)-O(3S)1,460(4)S(1S)-O(2S)1,461(4)
S(1S)-O(1S)1,463(4)S(1S)-O(4S)1,474(4)
O(1W)-H(1WB),8500 O(1W)-H(1WA)0,8499
O(2W)-H(2WA)0,8499O(2W)-H(2WB)0,8500
O(3W)-H(3WB)0,8499O(3W)-H(3WA)0,8500
The valence angleAboutThe valence angleAbout
N(2)-Fe(1)-N(1)to 121.6(2)
N(2)-Fe(1)-S(1)106,61(16)N(1)-Fe(1)-S(1)104,59(15)
N(2)-Fe(1)-S(1)#1108,03(16)N(1)-Fe(1)-S(1)#1108,06(15)
S(1)-Fe(1)-S(1)#1107,07(5)N(2)-Fe(1)-Fe(1)#1120,07(16)
N(1)-Fe(1)-Fe(1)#1118,22(15)S(1)-Fe(1)-Fe(1)#153,68(4)
S(1)#1-Fe(1)-Fe(1)#153,40(4) C(1)-S(1)-Fe(1)106,78(17)
C(1)-S(1)-Fe(1)#1108,44(18)Fe(1)-S(1)-Fe(1)#172,93(5)
O(1)-N(1)-Fe(1)171,4(4)O(2)-N(2)-Fe(1)173,7(4)
C(2)-N(3)-H(3NA)114,5C(2)-N(3)-H(3NC)111,2
H(3NA)-N(3)-H(3NC)109,7C(2)-N(3)-H(3NB)108,8
H(3NA)-N(3)-H(3NB)to 112.4H(3NC)-N(3)-H(3NB)of 99.1
C(2)-C(1)-S(1)109,4(4)C(2)-C(1)-H(1A)109,8
S(1)-C(1)-H(1A)109,8C(2)-C(1)-H(1B)109,8
S(1)-C(1)-H(1B)109,8H(1A)-C(1)-H(1B)to 108.2
N(3)-C(2)-C(1)110,0(4)N(3)-C(2)-H(2A)109,7
C(1)-C(2)-H(2A)109,7N(3)-C(2)-H(2B)109,7
C(1)-C(2)-H(2B)109,7H(2A)-C(2)-H(2B)to 108.2
N(1')-Fe(1')-N(2')121,3(2)N(1')-Fe(1')-S(1')#2103,85(15)
N(2')-Fe(1')-S(1')#2107,69(16)N(1')-Fe(1')-S(1')110,10(15)
N(2')-Fe(1')-S(1')106,37(15)S(1')#2-Fe(1')-S(1')106,74(5)
N(1')-Fe(1')-Fe(1')#2119,25(15)N(2')-Fe(1')-Fe(1')#2119,39(15)
S(1')#2-Fe(1')-Fe(1')#253.47 USD(4)S(1')-Fe(1')-Fe(1')#253,27(4)
C(1')-S(1')-Fe(1')#2105,82(17)C(1')-S(1')-Fe(1')107,88(18)
Fe(1')#2-S(1')-Fe(1')73,26(5)O(1')-N(1')-Fe(1') 171,2(4)
O(2')-N(2')-Fe(1')172,4(4)C(2')-N(3')-H(3NF)110,1
C(2')-N(3')-H(3ND)110,3H(3NF)-N(3')-H(3ND)102,8
C(2')-N(3')-H(3NE)104,9H(3NF)-N(3')-H(3NE)111,2
H(3ND)-N(3')-H(3NE)117,5C(2')-C(1')-S(1')110,3(4)
C(2')-C(1')-H(1'A)109,6S(1')-C(1')-H(1'A)109,6
C(2')-C(1')-H(1'B)109,6S(1')-C(1')-H(1'B)109,6
H(1'A)-C(1')-H(1'B)108,1N(3')-C(2')-C(1')110,6(4)
N(3')-C(2')-H(2'A)109,5C(1')-C(2')-H(2'A)109,5
N(3')-C(2')-H(2'B)109,5C(1')-C(2')-H(2'B)109,5
H(2'A)-C(2')-H(2'B)108,1O(3S)-S(1S)-O(2S)109,4(2)
O(3S)-S(1S)-O(1S)109,2(2)O(2S)-S(1S)-O(1S)109,7(3)
O(3S)-S(1S)-O(4S)110,9(2)O(2S)-S(1S)-O(4S)108,8(2)
O(1S)-S(1S)-O(4S)108,7(2)H(1WB)-O(1W)-H(1WA)99,0
H(2WA)-O(2W)-H(2WB)117,2H(3WB)-O(3W)-H(3WA)95,4
Operations of symmetry: #1-x,-y+1,-z+2 #2-x+1,-y,-z+2

1,489(5) 169,1(3)td align="justify"> O(8)-C(8)-C(7)
Table 4
Interatomic distances and valence angles in the structure of the complexII
CommunicationǺCommunicationǺ
Fe(1)-N(2)1,666(3)Fe(1)-N(1)1,669(3)
Fe(1)-S(2)2,2545(10) Fe(1)-S(1)2,2620(9)
Fe(1)-Fe(2)2,7088(6)Fe(2)-N(4)1,676(3)
Fe(2)-N(3)1,680(3)Fe(2)-S(2)2,2564(9)
Fe(2)-S(1)2,2601(11)S(1)-C(1)1,873(3)
S(2)-C(6)1,868(4)N(1)-O(1)1,170(4)
N(2)-O(2)1,172(4)N(3)-O(3)1,162(4)
N(4)-O(4)1,165(4)N(5)-C(2)1,496(5)
N(5)-H(5NC)0,8500N(5)-H(5NB)0,8500
N(5)-H(5NA)0,8498O(5)-C(3)1,216(4)
O(6)-C(3)1,304(4)O(6)-H(6O)0,8500
N(6)-C(7)N(6)-H(6NC)0,8500
N(6)-H(6NB)0,8500N(6)-H(6NA)0,8499
O(7)-C(8)1,198(4)O(8)-C(8)1,322(4)
O(8)-H(8O)0,8501C(1)-C(5)1,530(5)
C(1)-C(4)1,541(5)C(1)-C(2)1,554(5)
C(2)-C(3)1,529(5)C(2)-H(2A)1,0000
C(4)-H(4A)0,9800C(4)-H(4B)0,9800
C(4)-H(4C)0,9800C(5)-H(5A)0,9800
C(5)-H(5B)0,9800C(5)-H(5C)0,9800
C(6)-C(10)1,512(5)C(6)-C(9)1,538(5)
C(6)-C(7)1,557(5)C(7)-C(8)1,531(5)
C(7)-H(7A)1,0000C(9)-H(9A)0,9800
C(9)-H(9B)0,9800C(9)-H(9C)0,9800
C(10)-H(10A)0,9800C(10)-H(10B)0,9800
C(10)-H(10C)0,9800O(1W)-H(1WB)0,8500
O(1W)-H(1WA)0,8500O(1S)-S(3')1,455(5)
O(1S)-S(3)1,492(5)S(3)-O(4S)1,481(3)
S(3)-O(3S)1,482(3)S(3)-O(2S)1,486(3)
S(3')-O(3S')1,481(3)S(3')-O(4S')1,481(3)
S(3')-O(2S')1,483(3)O(2W)-H(2WB) 0,8501
O(2W)-H(2WA)0,8501O(3W)-H(3WB)0,8500
O(3W)-H(3WA)0,8500O(4W)-H(4WB)0,8500
O(4W)-H(4WA)0,8500
The valence angleAboutThe valence angleAbout
N(2)-Fe(1)-N(1)113,95(14)N(2)-Fe(1)-S(2)111,94(11)
N(1)-Fe(1)-S(2)106,21(10)N(2)-Fe(1)-S(1)106,47(11)
N(1)-Fe(1)-S(1)111,79(10)S(2)-Fe(1)-S(1)106,29(4)
N(2)-Fe(1)-Fe(2)122,38(10)N(1)-Fe(1)-Fe(2)123,67(10)
S(2)-Fe(1)-Fe(2)53,13(2)S(1)-Fe(1)-Fe(2)53,17(3)
N(4)-Fe(2)-N(3)113,40(14)N(4)-Fe(2)-S(2)111,37(11)
N(3)-Fe(2)-S(2)104,79(11)N(4)-Fe(2)-S(1)108,57(11)
N(3)-Fe(2)-S(1)112,19(11)S(2)-Fe(2)-S(1)106,29(4)
N(4)-Fe(2)-Fe(1)123,85(10)N(3)-Fe(2)-Fe(1)122,68(10)
S(2)-Fe(2)-Fe(1)53,06(3)S(1)-Fe(2)-Fe(1)53,23(3)
C(1)-S(1)-Fe(2)112,90(12)C(1)-S(1)-Fe(1)116,61(11)
Fe(2)-S(1)-Fe(1)73,60(3)C(6)-S(2)-Fe(1)114,39(11)
C(6)-S(2)-Fe(2)116,40(11)Fe(1)-S(2)-Fe(2)73,81(3)
O(1)-N(1)-Fe(1)is 165.8(3)O(2)-N(2)-Fe(1)169,8(3)
O(3)-N(3)-Fe(2)O(4)-N(4)-Fe(2)164,4(3)
C(2)-N(5)-H(5NC)116,0C(2)-N(5)-H(5NB)113,8
H(5NC)-N(5)-H(5NB)102,7C(2)-N(5)-H(5NA)109,8
H(5NC)-N(5)-H(5NA)105,9H(5NB)-N(5)-H(5NA)108,1
C(3)-O(6)-H(6O)110,8C(7)-N(6)-H(6NC)109,8
C(7)-N(6)-H(6NB)113,5H(6NC)-N(6)-H(6NB)101,4
C(7)-N(6)-H(6NA)102,2H(6NC)-N(6)-H(6NA)111,0
H(6NB)-N(6)-H(6NA)119,0C(8)-O(8)-H(8O)111,5
C(5)-C(1)-C(4)109,5(3)C(5)-C(1)-C(2)110,4(3)
C(4)-C(1)-C(2)109,5(3)C(5)-C(1)-S(1 114,6(3)
C(4)-C(1)-S(1)to 105.3(2)C(2)-C(1)-S(1)107,2(2)
N(5)-C(2)-C(3)to 107.7(3)N(5)-C(2)-C(1)110,9(3)
C(3)-C(2)-C(1)to 114.7(3)N(5)-C(2)-H(2A)107,8
C(3)-C(2)-H(2A)107,8C(1)-C(2)-H(2A)107,8
O(5)-C(3)-O(6)125,2(3)O(5)-C(3)-C(2)122,4(3)
O(6)-C(3)-C(2)to 112.4(3)C(1)-C(4)-H(4A)109,5
C(1)-C(4)-H(4B)109,5H(4A)-C(4)-H(4B)109,5
C(1)-C(4)-H(4C)109,5H(4A)-C(4)-H(4C)109,5
H(4B)-C(4)-H(4C)109,5C(1)-C(5)-H(5A)109,5
C(1)-C(5)-H(5B)109,5H(5A)-C(5)-H(5B)109,5
C(1)-C(5)-H(5C)109,5H(5A)-C(5)-H(5C)109,5
H(5B)-C(5)-H(5C)109,5C(10)-C(6)-C(9)109,7(3)
C(10)-C(6)-C(7)113,9(3)C(9)-C(6)-C(7)109,5(3)
C(10)-C(6)-S(2)115,0(2)C(9)-C(6)-S(2)105,0(2)
C(7)-C(6)-S(2)103,2(2)N(6)-C(7)-C(8)109,2(3)
N(6)-C(7)-C(6)of 112.8(3)C(8)-C(7)-C(6)113,1(3)
N(6)-C(7)-H(7A)107,1C(8)-C(7)-H(7A)107,1
C(6)-C(7)-H(7A)107,1O(7)-C(8)-O(8)125,7(3)
O(7)-C(8)-C(7)122,6(3)111,7(3)
C(6)-C(9)-H(9A)109,5C(6)-C(9)-H(9B)109,5
H(9A)-C(9)-H(9B)109,5C(6)-C(9)-H(9C)109,5
H(9A)-C(9)-H(9C)109,5H(9B)-C(9)-H(9C)109,5
C(6)-C(10)-H(10A)109,5C(6)-C(10)-H(10B)109,5
H(10A)-C(10)-H(10B)109,5C(6)-C(10)-H(10C)109,5
H(10A)-C(10)-H(10C)109,5H(10B)-C(10)-H(10C)109,5
H(1WB)-O(1W)-H(1WA)118,4O(4S)-S(3)-O(3S)109,3(4)
O(4S)-S(3)-O(2S)113,3(4)O(3S)-S(3)-O(2S)110,0(4)
O(4S)-S(3)-O(1S)111,1(4)O(3S)-S(3)-O(1S) 111,3(3)
O(2S)-S(3)-O(1S)of 101.7(3)O(1S)-S(3')-O(3S')106,5(3)
O(1S)-S(3')-O(4S')to 112.2(3)O(3S')-S(3')-O(4S')108,1(4)
O(1S)-S(3')-O(2S')112,6(4)O(3S')-S(3')-O(2S')112,0(4)
O(4S')-S(3')-O(2S')to 105.3(4)H(2WB)-O(2W)-H(2WA)114,8
H(3WB)-O(3W)-H(3WA)byr111.4H(4WB)-O(4W)-H(4WA)97,7

The molecular structure of complexesIandIIpresented on figures 1 and 3, respectively. According to PCAIone molecule of sulfate salt of the dication [Fe2(S(CH)2NH3)2(NO)4]2+crystallizes with 2.5 molecules of water. In the crystal structure ofItwo independent dication (both of centrosymmetric), in which the iron atoms are bound to two nitrogen atoms nitrosyl ligands and two atoms μ-S protonated cysteamine ligands. The latter are characterized by antiperiplanar conformation with torsion the corners SCCN, equal 169,1° and 179,8°. The iron atoms in addition to Fe(1)-Fe(1A) is in a slightly distorted tetrahedral configuration with a maximum deviation angle of NO-Fe-NO to 121,3(2)°. Although the geometry of two independent dications are almost identical, the distance Fe...Fe in them a little different and equal 2,672 (1) and 2,682 (1) Å. The deviation from linearity relations of the Fe-NO on average about 10°. On the one hand, it can be the result only of the geometry of four-membered rings of Fe2S2. It should be noted that the distance Fe...Fe in these complexes is quite sensitive to the effects of crystal packing, and, for example, in the above-mentioned iodide equal 2,715 (1) Å.

On one molecule of the dication complexIIhave sulfate anion and 5 water molecules. The dication has having a center of inversion dimer binuclear structure, where two tetrahedrally coordinated iron atom linked to two NO groups and two penicillamine Mililani. Between the iron atoms linked through a fragment of the μ-S penicillamine ligands. The distance Fe(1)-Fe(2) is 2,7088(6).

Summing up the analysis of the geometry of the dication, we can conclude that the structure of the complexesIandIIrefers to the structural type esters red salt Russa [N.A. Sanin, S.M. Aldoshin, Izv. An. Ser. Chem., 2004,11, 2326].Taking into account the fact that the complexesIandI are diamagnetic, the relatively small distance Fe...Fe may indicate the existence of a relationship between iron atoms.

The crystal structures of complexesIandIIpresented in figure 2 and 4, respectively.

The study of NO-donor activity of complexesIandII

For determination of NO, generated by complexes I and II in water, used the touch electrode amiNO-700” system “inNO Nitric Oxide Measuring System” (Innovative Insruments, Inc., Tampa, FL, USA). The NO concentration was fixed for ~1600 seconds (in increments of 0.2 sec) c concentration of NO donor (0.1 Microm). For calibration of the electrochemical sensor used standard aqueous solution of NaNO2(100 Microm), which was added to a mixture of 0.12 m KI and 2 ml of 1M H2SO4in 18 ml of water. All experiments were performed in aerobic solutions at 25°C. the pH of the solutions was measured using membrane pH meter HI 8314” (HANNA instruments, Germany).

It is established that complexesIandIIgenerate NO in aerobic aqueous solutions spontaneously, i.e. in the absence of chemo-, photo - or enzymatic activation (figure 5, 6). Unlike binuclear nitrosyl iron complexes with aromatic Mililani, complexesIandIIallocate NO prolongirovanne.

The study of cytotoxicity of water-soluble iron complexesIandIItumor is x human cells in vitro

The object of the studies used cells erythroblastosis human leukemia of K 562 line cells and non-small cell lung cancer A. Cells were grown in RPMI1640 medium containing 10% fetal calf serum at 37°C, 5% CO2and 100% humidity.

Testing the cytotoxic activity was performed by standard MTT test using a 96-well plate. The density of cells was 5000 cells per well. Cells C and A incubated with complexesIandIIat concentrations of 1 μm, 10 μm or 100 μm for 72 hours.

The MTT test is based on the ability of the dehydrogenases of living cells to restore unpainted MTT reagent in the form of blue crystals formazan, soluble in DMSO. The optical absorption of the colored solutions of DMSO was measured on an automatic counter. Cytotoxic activity was evaluated in the % survival of the cells and was calculated by the formula: (mean value of optical absorption in the experimental samples/average absorption in control) x 100. IR50was determined by curve "dose-effect".

Study of the cytotoxic activity nitrosyl iron complexesIandIIon K562 cells and A showed a direct dependence of the survival rate of cells on the concentration ofIandII. Values IR50forIwas 50 μm, forI 100 microns.

The study apoptotic activity of water-soluble iron complexes on tumor cells

Method for determining the activity of caspases is based on the use fluorochrome inhibitors of caspases (FLICA). These inhibitors are able to penetrate into the cell and are not cytotoxic. Inside the cell FLICA-inhibitor covalently binds to the active caspase. For reagents (whales), fluorescent in the green region of the spectrum, use the labeled inhibitor of caspase - carboxyfluorescein fluoromethyl ketone peptide.

For testing of apoptotic cells in the experiments was used liofilizirovanny reagent FAM-DEVD-FMK for determination of caspase-3 and caspase-7. Adding to the population of cells FAM-DEVD-FMK enters each cell and covalently binds to the large subunit of active Casanova of heterodimer and thus inhibits the enzymatic activity. The reagent is covalently linked to the enzyme remains inside the cell. Part of the unbound reagent diffuses out of the cell and can be removed by washing. The remaining green fluorescent signal is directly dependent on the number of active kaspasky enzymes that were present in the cell at the time of adding the reagent.

The working reagent solution was obtained by adding to liofilizirovannom reagent FAM-DEVD-FMK 50 ál DMSO (h t the article), and then 200 μl of phosphate buffer solution to obtain the 30-s test.

To remove unbound reagent FAM-DEVD-FMK use of available ICT firm 10x Buffer for washing cells. For the experiments we used diluted with distilled water 1x buffer.

For experiments cells were scattered in the vials with a density of 500 cells per ml of Induced cells were obtained by incubation for 72 hours with nitrosyl complexesIandIIat equimolar concentrations of 50 µm, close to the IC50for all complexes.

As a negative control was used neindutsirovannom unpainted cells and FLICA-cells, and also induced unpainted cells.

At the end of the period of incubation, the cells were centrifuged for 5 minutes at 1200 rpm at room temperature (RT). Sediment cells resuspendable in 300 μl of medium was added 10 μl of 30× FLICA and incubated for 1 hour at 37°C and 5% CO2. After incubation, the samples were centrifuged, supernatant was removed. Then double-washed precipitate in cells 1× Buffer (2 ml and 1 ml). Sediment cells resuspendable 900 μl of 1× Buffer. Selected aliquots of the samples induced and reinducing cells were added 2 μl of ready to use proprietary reagent propedy iodide (PI) and incubated at RT for 10 minutes for okresu is of dead cells.

The fluorescence of all samples of cells were analyzed on a liquid cytometer BACKTON Dikenson Fascalibur using a laser with a wavelength of 488 nm. The results were obtained in the form of the distribution of fluorescent signals stained cells in channel FL1 (green FLICA)and FL3 (red region PI). In the lower left quadrant are defined by living cells; in the lower right - FLICA positive cells (FLICA+)in top left-painted PI cells; in the upper right-hand - painted PI and FLICA+cells. Graphic images of cells in the channels received using the Dot-Raft. Quantification of results was performed using the program WinMDA version 2/8 and were expressed as the content of painted and unpainted cells relative to the total number of cells in %.

Investigated nitrosyl complexIdemonstrated ability to initiate activity of caspase 3 and 7 and, thus, to induce apoptosis in cells erythroblastosis human leukemia of K 562 line cells. In the presence of complexIregistered 77% of cells in apoptosis (7).

1. Binuclear nitrosyl cation complex of iron with natural aliphatic Mililani General formula [Fe2(SR)2(NO)4]SO4where R represents an aliphatic ligands of natural origin.

2. Binuclear nitrosyl cation idea is with iron according to claim 1, where R represents a group probably facilitates or penicillamine.

3. A method of obtaining a binuclear cationic nitrosyl iron complexes according to claim 1, consisting in that an aqueous solution of iron sulfate (II) is treated with an appropriate water-soluble tiamina in the presence of gaseous NO in an acidic environment.

4. The method of claim 3, wherein the process is conducted at room temperature, preferentially at 18-25°C.

5. The method of claim 3, wherein the process is conducted in an oxygen-free atmosphere.

6. How to claim 3, characterized in that as thiamine use the group probably facilitates or penicillamine.

7. The donor of nitrogen monoxide, which represents a binuclear nitrosyl cation complex of iron according to claim 1.

8. Inducer of apoptosis of tumor cells, which represents a binuclear nitrosyl cation complex of iron according to claim 1.

9. The use of binuclear cationic nitrosyl iron complex according to claim 1 as an antitumor drug.

10. The use of binuclear cationic nitrosyl iron complex according to claim 1 for receiving anticancer drugs.

11. Pharmaceutical composition comprising an effective amount binuclear cationic nitrosyl iron complex according to claim 1 and a pharmaceutically acceptable carrier, as an antitumor agent.

12. Pharm is citiesa the composition according to claim 11, where the pharmaceutically acceptable carrier used protonotariou environment.

13. The pharmaceutical composition according to item 12, where as protonotaria environment using water, saline solution, water-soluble biopolymers.

14. The pharmaceutical composition according to claim 11, where binuclear nitrosyl cation complex of iron is present in the amount of 50-100 microns.

15. The set used for the treatment of cancer, comprising (1) a pharmaceutical composition comprising a binuclear nitrosyl cation complex of iron according to claim 1, in a sealed package and (2) auxiliary components.



 

Same patents:

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

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing mono-imine compounds of formula where radicals assume values given in claim 1 of the invention, involving reaction of a dicarbonyl compound with aniline in an aliphatic non-aromatic solvent. The invention also describes an asymmetric iron complex of formula , where radicals assume values given in claim 3 of the invention, as well as a catalyst system for polymerisation of olefins.

EFFECT: invention describes mono-imine compounds having electron-attracting substitutes in the ortho-position and asymmetric bis(imino)-compounds and asymmetric iron compounds obtained from said compounds, and use of the complexes in olefin polymerisation.

7 cl, 11 ex, 1 tbl

FIELD: information technology.

SUBSTANCE: article with an image includes a substrate having a masked or concealed protective image on at least part thereof, which reflects less than 50% of radiation at wavelength 800-900 nm. The protective image contains an infrared radiation absorbing compound selected from:

or salt or polymer thereof, where M is a metal selected from iron, cobalt, nickel, aluminium, scandium, chromium, vanadium, titanium, manganese and lanthanide. R1 is selected from hydrogen, phosphonate, sulphonate, nitro, halogen, cyano, thiocyano, thioalkyl, thioaryl, alkyl, alkoxy, aryl, aryloxy, amine, substituted amines and substituted aryl. One of R2 and R3 is oxygen and the other is NO; n is a number corresponding to half the coordination number of metal M; each of L and L' independently denotes a ligand which forms a complex with metal M, and y is a number which corresponds to the coordination number of metal M. The infrared radiation absorbing compound does not form an intensely coloured protective image, and the protective image is pale, colourless or tinted. The invention also discloses a method of making the article with the image, using the compound and a method of authenticating the said article.

EFFECT: obtaining a protective image which can reflect less than half the light at 800-900 nm, and a protective image which is not intensely coloured.

14 cl, 35 ex, 4 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to use of a tetranitrosyl complex of iron with thiophenol of formula [Fe2(SC6H5)2(NO)4] as an anti-tumour medicinal agent in order to prepare a medicinal agent for treating oncological diseases. The invention also relates to a pharmaceutical composition and a set containing said complex.

EFFECT: invention ensures high efficiency of treatment.

8 cl, 1 tbl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to complex compound of iron with carbohydrate. Complex compound, in addition to iron (III) contains iron (II). Per cent of iron (II) in terms of total amount of iron in complex compound constitutes, at least, 2 wt %. Carbohydrate is selected from group, consisting of natural carbohydrates or synthetic derivatives of carbohydrates, such as starch, hydrolysed starches, dextrins, in particular such as maltodextrin, maltose syrup, glucose syrup, cyclodextins, dextrans, saccharides. Per cent of carbohydrate in terms of complex compound of iron with carbohydrate constitutes from 10 to 80 wt %. Method of obtaining complex compound of iron with carbohydrate includes the following stages. Water solution of suspension of carbohydrate is prepared. After that, iron (III) salts are added at constant pH value within the range from 7 to 13. Iron (III) salt is selected from group, consisting of iron (III) chloride and iron (III) sulfate. Water solution or suspension is heated with further cooling. Formed complex compound of iron with carbohydrate if isolated. Precipitation of complex compound of iron with carbohydrate is performed by addition of one or more alcohols, such as ethanol or propanol. Obtained complex compound is used for production of medication for treatment of iron-deficient anemias, which is well absorbed in organism in case of peroral intake.

EFFECT: iron, included into composition of medication is better resorbed and better tolerated by organism than pure iron preparations.

26 cl, 1 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing trihydrate of iron (II) aminobutyrate-riboflavinate Fe(C4H8O2N)(C17H19O6N4) 3H2O, which can be used as a biologically active additive which enhances growth and development of animals. The method is implemented by mixing an aqueous suspension of riboflavinate and iron (II) gamma-aminobutyrate in equimolar ratio and temperature 70-80°C.

EFFECT: high output while cutting the time for implementing the method.

1 cl, 3 ex

FIELD: agriculture.

SUBSTANCE: invention relates to a novel compound - dihydrate of iron (II) hydroascorbate riboflavinate

(II) Fe(C6H7O6)(C17H19O6N4)2H2O which is used as an additive.

EFFECT: improved development and growth of chickens.

1 cl, 1 ex, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to 1-alkenylimidazole derivatives of general formula 1 , where R is vinyl, allenyl or isopropenyl, R1 is hydrogen or methyl, E is Zn (II) or Fe(III), An is chlorine or acetate, n equals 1, 2 or 4, except compounds where R is vinyl, R1 is hydrogen, E is Zn(II), An is chlorine or acetate and n equals 2.

EFFECT: compounds have antihypoxic properties for different types of hypoxia.

5 ex, 8 tbl

FIELD: process engineering.

SUBSTANCE: invention relates to complete methane oxidation catalysts and can be used in industries using diesel fuel. Invention covers complete methane oxidation catalysts based on strontium hexaferrites of the following composition: SrMnxFe12-xO19, where x=0, 1, 2, 6. Proposed method comprises settling catalyst components with the help of NH4HCO3 solution at constant pH equal to (7.1 to 8.0) and temperature not lower than 70°C with subsequent stages of filtration, rinsing, drying and roasting. Proposed method comprises also the stage of heat treatment at 800° to 1000° C and is realised in the presence of above described catalysts.

EFFECT: high degree of methane conversion at relatively low temperatures.

6 cl, 2 tbl, 14 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a polyolefin synthesis method and more specifically to a polyethylene synthesis method. Polyethylene is a copolymer of ethylene with 1-alkenes. The invention also relates to polyethylene synthesis catalyst systems. The catalyst system is a mixture of metallocenes: hafnocene and an iron-based complex, an activating compound and a support. The invention also relates to films made from polyethylene and packets made from the said films.

EFFECT: disclosed catalyst system enables production of polyethylene with given molecular weight distribution in a single reactor.

16 cl, 3 tbl, 3 ex

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

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds of general formula (I) where the bond b represents a double bond; X represents -S-; each Z1 and Z3 independently represents a direct bond, -N(R5) - or - (CH2)q; Z2 represent -C(O)- or -C(S)-; m represents an integer equal to 1; n represents an integer equal to 1; each of q independently represents an integer varying within 1 to 4; R0 represents hydrogen, halogen, hydroxy, unsubstituted C1-C3alkyl or unsubstituted C1-C3alkoxy; R1 is independently selected from a group consisting of halogen, optionally substituted C1-C3alkyl, -R6OR7, -R6N(R7)2, -R6C(O)R7, -R6C(O)OR7, -R6C(O)N(R7)R9N(R7)2, -R6OC(O)R8, -R6C(O)N(R7)2 or -R6OR9N(R7)2; R2 represents hydrogen; R4 is selected from a group consisting of morpholine, isoxazolyl, thiazolyl, oxazolyl, benzisoxazolyl, benzothiazolyl, dioxynyl, dioxolyl, and optionally substituted phenyl. Also, the invention refers to pharmaceutically acceptable salts of the compounds of formula (I) and to a pharmaceutical composition exhibiting antiproliferative activity and containing the compounds of formula (I).

EFFECT: preparing the compounds of formula (I) exhibiting antiproliferative activity.

21 cl, 11 dwg, 5 tbl, 19 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: there are described new camptothecin derivatives, i.e. 4,5-dihydrotriazolyl [5,4-c]16a-deoxocamptothecin and triazolyl[5,4-c]16a-deoxocamptothecin showing anticancer activity, a pharmaceutical composition containing them and applying them as anticancer drugs.

EFFECT: preparing the anticancer pharmaceutical composition.

5 cl, 11 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds of formula (I) exhibiting the properties of proteinkinase Plk1 inhibitors. In formula (I): the ring A is a 5-member heteroaryl ring which contains atoms N as heteroatoms where the ring is optionally substituted by C1-6alkyl; X1 is NR8-; R1 is H, C1-10 aliphatic, C3-10 cycloaliphatic, phenyl, 5-6- member heteroaryl group containing atom S, N as a heteroatom where said R1 is optionally substituted by 0-5 J1; each R2 and R3, is independently H, C1-10 aliphatic group; R4 is C3-10 cycloaliphatic group; R8 is H, -C(O)OR; each J1 is independently C1-6 halogenalkyl, halogen, Q or -Z-Q; each Z is independently C1-6 aliphatic group where 0-3 -CH2- links in the specified C1-6 aliphatic group are optionally replaced by -NR-, -O-, -C(O)-, where any of the unsubstituted -CH2- links in the specified C1-6 aliphatic group is optionally substituted by 0-2 Jz ; each Q is independently H, C1-6 aliphatic group, 3-8- member aromatic or non-aromatic monocyclic ring having 0-3 heteroatoms independently selected from O and N where each Q is independently and optionally substituted by 0-2 JQ; Jz is halogen; JQ is -M; each M is independently H, C1-6 aliphatic group, halogen; each R is independently H or unsubstituted C1-6 aliphatic group.

EFFECT: preparing the pharmaceutical composition and method for inhibiting proteinkinase Plk1 activity.

31 cl, 43 ex, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds selected from a group consisting of compounds of formula: and or to their pharmaceutically acceptable salts. Also, the invention refers to a pharmaceutical composition, as well as to using at least one compound under cl.1 and/or its pharmaceutically acceptable salts.

EFFECT: preparing new biologically active compounds which exhibit the properties of cycline-dependent kinase inhibitors.

11 cl, 86 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to imidazopyridines of formula I

and to their pharmaceutically acceptable salts where Z1 represents CR1; R1 represents H; R1' represents H; Z2 represents CR2; Z3 represents CR3 or N; R2 and R3 are independently selected from H, halogen; R4 represents H; Y represents W-C(O)-; W represents or , R5 represents H; X1 is selected from R11' and -OR11'; each R11 independently represents H, C1-C12alkyl, C2-C8alkenyl; X4 represents , R6 represents H, halogen, cyclopropyl or -(CR19R20)n-SR16; R6 represents H, halogen; p represents 0, 1, 2 or 3; n represents 0, 1 or 2; where each specified alkyl in R11 is independently substituted by one or two groups independently selected from halogen, -(CR19R20)nOR16 and R21; each R16 independently represents H, C1-C12alkyl; R19 and R20 are independently selected from H, C1-C12alkyl; R21 represents cyclopropyl.

EFFECT: invention refers to a pharmaceutical composition for treating hyperproliferative disorder.

10 cl, 25 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds of formula:

in which: X represents one of the following groups: - a phenyl group optionally substituted by one or more groups, optionally selected from one of the following atoms and groups: halogen, (C1-C6)alkoxy, (C1-C6)alkyl, (C3-C7)cycloalkyl(C1-C6)alkyl, (C3-C7)cycloalkyl(C1-C6)alkoxy, NRaRb, R1 represents hydrogen atom, halogen, (C1-C6)alkoxy, (C1-C6)alkyl, (C3-C7)cycloalkyl(C1-C6)alkyl, (C3-C7)cycloalkyl(C1-C6)alkoxy, amino, the group NRcRd; with the alkyl and alkoxy groups being optionally substituted by one or more halogens, hydroxy, amino or (C1-C6)alkoxy, R2 represents one of the following groups: -hydrogen atom, - (C1-C6)alkyl group optionally substituted by one or more groups optionally substituted by hydroxy, halogen, amino, the group NRaRb, the phenyl group, the - (C1-C3)alkoxygroup optionally substituted by one or more groups independently selected from hydroxy, halogen, amino, the group NRaRb, - (C3-C7)cycloalkyl((C1-C6)alkyl, - (C3-C7)cycloalkyl(C1-C6)alkoxy, -(C2-C6)alkenyl, - (C2-C6)alkinyl, - the group -CO-R5,- the group -CO-NR6R7,- the group -CO-O-R8,- the group -NR9-CO-R10,- the group -NR11R12,- halogen atom,- the cyanogroup,- the phenyl group optionally substituted by one or more groups optionally selected from the following atoms and groups: halogen, (C1-C6)alkoxy, NRaRb, -CO-R5, -CO-NR6R7, -CO-O-R8, (C3-C7)cycloalkyl(C1-C6)alkyl, (C3-C7)cycloalkyl(C1-C6)alkoxy, (C1-C6)alkyl group optionally substituted by one or more hydroxy groups or NRaRb R3 represents hydrogen atom, (C1-C6)alkyl, (C1-C6)alkoxy or halogen atom, R4 represents hydrogen atom, (C1-C4)alkyl, (C1-C4)alkoxy or fluorine atom, R5 represents hydrogen atom, the phenyl group or (C1-C6)alkyl, R6 and R7, the same or different represents hydrogen atom or (C1-C6)alkyl, or together with nitrogen atom forms a 4-7-member cycle optionally containing the other heteroatom selected from N, O or S,R8 represents (C1-C6)alkyl, R9 and R10, the same or different, represent hydrogen atom or (C1-C6)alkyl, R11 and R12, the same or different, represent (C1-C6)alkyl, or together with nitrogen atom form a 4-7-member cycle optionally containing the other heteroatom selected from N, O or S, Ra and Rb independently represent hydrogen atom, (C1-C6)alkyl or together with nitrogen atom form a 4-7-member cycle, Rc represent hydrogen atom, and Rd represents (C1-C6)alkyl and at least one of the substitutes R1, R2, R3 and R4 are different from hydrogen; and when R3 means methyl, X is unsubstituted; when R1 means methyl, X is unsubstituted; when R2 means chlorine, X is other than parafluorophenyl; in the form of a base or an acid addition salt. The invention also refers to the compounds selected from the group, to a drug, to a pharmaceutical composition, as well as to application of the compounds of formula (I) by any of cl. 1-4.

EFFECT: preparing new biologically active compounds for treating or preventing the diseases associated with nuclear receptor NOT.

13 cl, 18 ex, 2 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a new compound 4-(1-(4-(4-methoxyphenylthio)-2,5-dioxo-2,5-dihydro-1H-pyrro-3-yl)-1H-indol-3-yl)butyl carbamimidothioate (the compound "ЛХТА-1833") which can be used in therapy of colon cancer, breast cancer, melanoma, leucosis.

EFFECT: preparing the new compounds for treating cancer.

2 cl, 2 tbl, 3 dwg, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to 4-arylcumarine derivatives and a based drug which can be used for treating tumours of general formula

, where A means a group , where CnHm represents residual fatty acids of saturate and olefin series, n represents number 13, 15 or 17, m represents number 27, 31 or 33, while R1, R2, R3 represent hydrogen atoms and/or methoxygroup OCH3. The drug contains said 4-arylcumarine derivatives contained in liposomes containing phosphatidylcholine, phosphatidylinositol and lipid oligopolysaccharide.

EFFECT: there are prepared the new oncology compounds and the new drug showing lesser systemic toxicity and greater membrane affinity.

12 cl, 9 ex, 1 tbl, 3 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to pharmaceutical industry, in particular to composition for prevention and treatment of cancer. Pharmaceutical composition for prevention and treatment of cancer, which contains one or more components selected from group, consisting of cell line obtained from Taxus cambium or procambium with specified characteristics. Functional food product for prevention or improvement of state in case of cancer, which contains one or more components selected from group, consisting of cell line obtained from Taxus cambium or procambium with specified characteristics.

EFFECT: composition and product efficiently induce death of cancer cells, have minimal side effects.

11 cl, 22 dwg, 4 tbl, 8 ex

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

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