Agent for treating body iron overload, or haemochromatosis

FIELD: medicine.

SUBSTANCE: method involves chelation of ferric (III) ions both in vivo, and in vitro with using 2-ethyl-6-methyl-3-hydroxypyridine succinate (Mexidol) to produce a stable stained complex having a high molecular weight and a complex (organic-inorganic) composition not identified among the known compounds.

EFFECT: high effectiveness of the chelation process and low toxicity of Mexidol that makes it promising for biomedical application.

4 dwg, 3 tbl, 3 ex

 

The invention belongs to the field of medicine, namely to new means of chelation of metal ions, mainly iron, which can be used in the treatment of an overload of iron in the body or in hemochromatosis.

Iron is a vital element that is part of hemoglobin, myoglobin, seminovich enzymes, but at th same time, it can increase the production of highly reactive and toxic free oxygen radicals, especially hydroxyl radicals, thereby stimulating oxidative damage [Iron and Heme Metabolism [Electronic resource]. - Search regimen http:/themedicfbiochemistrypage.org/heme-porphyrin.pgp]. Excess iron is associated with a number of diseases, disorders and conditions, because the person does not have physiological mechanisms aimed at removing an excess amount of this element [Brittenham G. M. Disorders of Iron Metabolism: Iron Deficiency and Overload / G. M. Brittenham // Hematology: Basic Principles and Practice / R. Hoffman, E. J. Benz, S. J. Shattil [et al.]. - [3rded.]. - New York: Churchill Livingstone, 2000. - P. 397-428; Fleming R. E. Iron overload in human disease / R. E. Fleming, P. Ponka // N. Engl. J. Med. - 2012. - Vol. 366, No. 4. - P. 348-359]

Hereditary hemochromatosis, a condition in which the body accumulates excess iron is one of the most common genetic diseases in humans. In the US, about one million people have signs of hemochromatosis, and about one in �adih ten people can carry the gene for this metabolic disorder [M. Franchini Recent advances in hereditary hemochromatosis / M. Franchini, D. Veneri // Ann. Hematol. - 2005. - Vol. 84, No. 6. - P. 347-352; Fowler C. Hereditary hemochromatosis: pathophysiology, diagnosis, and management / C. Fowler // Crit. Care Nurs. Clin. North. Am. - 2008. - Vol. 20, No. 2. - P. 191-201]. Hemochromatosis is characterized by excessive absorption of iron from food, its accumulation in the organs of the body and injury, including serious and even fatal health consequences as cirrhosis, liver cancer, heart failure, diabetes, impotence, and arthritis.

Clinical thalassemia (major and minor) is a hereditary disease characterized by impaired synthesis of hemoglobin, which leads to lower education and increased destruction of red blood cells. [Ip W. H. Diagnosis and prevention of thalassemia / H. W. Ip, C. C. So // Crit. Rev. Clin. Lab. Sci. - 2013. - Vol. 50, No. 6. - P. 125-141.]. In patients with β-thalassemia or resistant anemia (e.g., myelodysplastic syndrome) who get frequent or regular red blood cell transfusions in combination with increased iron absorption due to inefficient erythropoiesis, rapidly developing iron overload [Survival and complications in thalassemia / C. Borgna-Pignatti, M. D. Cappellini, P. De Stefano [et al.] // Ann. N. Y. Acad. Sci. - 2005. - Vol. 1054. - P. 40-47; Inati A. Recent advances in improving the management of sickle cell disease / A. Inati // Blood Rev. - 2009. - Vol. 23, suppl. 1. - S. 9-13].

Iron toxicity begins to occur when the load exceeds the binding capacity of blood and tissue and carried�asanee iron accelerates the formation of free radicals of oxygen, leading to peroxidative damage of cells. In the absence of iron chelation patients with regular blood transfusions inevitably subjected to a progressive deterioration of the functions of the pancreas, liver and heart as a result of iron overload, which usually occurs in the second decade of life [Thuret I. Post-transfusional iron overload in the haemoglobinopathies / I. Thuret // C. R. Biol. - 2013. - Vol. 336, No. 3. - P. 164-172].

Iron overload remains the leading cause of death among patients with thalassemia in industrialized countries. Despite progress in the treatment of such diseases, mortality among patients with β-thalassemia and sickle cell anemia in the United States is 3 times higher than among the total population [Morbidity and mortality in chronically transfused subjects with thalassemia and sickle cell disease: A report from the multi-center study of iron overload / E. B. Fung, P. Harmatz, M. Milet [et al.] // Am. J. Hematol. - 2007. - Vol. 82, No. 4. - P. 255-265]. For these patients, the only option to reduce the risk of serious complications is continuous therapy with iron chelators [Monitoring long-term efficacy of iron chelation therapy by deferiprone and desferrioxamine in patients with beta-thalassaemia major: application of SQUID biomagnetic liver susceptometry / R. Fischer, F. Longo, P. Nielsen [et al.] // Br. J. Haematol. - 2003. - Vol. 121, No. 6. - P. 938-948].

Although transfusion-dependent anemia are the main cause of iron overload, there are other clinical situations such complication, in particular, transplantation of hemopet�ical cells [N. S. Majhail Iron overload in hematopoietic cell transplantation / N. S. Majhail, H. M. Lazarus, J. L. Burns // Bone Marrow transplan. - 2008. - Vol. 41, No. 12. - P. 997-1003]. It is also known that the reduction of iron content in some organs and tissues, even in the context of its normal level in the body, can be useful in neurodegenerative diseases [The role of metals in neurodegenerative diseases / L. M. Sayre, G. Perry, C. S. Atwood [et al.] // Cell. Mol. Biol. - 2000. - Vol. 46. - P. 731-741; The transition metals copper and iron in neurodegenerative diseases / S. Rivera-Mancia, I. Perez-Neri, C. Rios [et al.] // Chem. Biol. Interact. - 2010. - Vol. 186, No. 2. - P. 184-199; Metal chelation as a potential therapy for Alzheimer's disease / M. P. Cuajungco, K. Y. Faget, X. Huang [et al.] // Ann. N. Y. Acad. Sci. - 2000. - Vol. 920. - P. 292-304], aging [A. S. Polla Iron as the malignant spirit in successful ageing / A. S. Polla, L. L. Polla, B. S. Polla // Ageing Res. Rev. - 2003. - Vol. 2, No. 1. - P. 25-37], cancer [Buss, J. L. The role of iron chelation in cancer therapy / J. L. Buss, F. M. Torti, S. V. Torti // Curr. Med. Chem. - 2003. - Vol. 10, No. 12. - P. 1021-1034] and other conditions and diseases associated with oxidative stress in the initiation of which a major role is played by iron ions [Free Radicals and Antioxidant Protocols / by ed. M. Rao Uppu, N. Subramanyam Murthy, William A. Pryor, L. Narasimham Parinandi // Series: Methods in Molecular Biology. - [2nded.]. - 2010. - Vol. 610. - 480 p.; Siddique A. Review article: the iron overload syndromes / A. Siddique, K. V. Kowdley // Aliment. Pharmacol. Ther. - 2012. - Vol. 35, No. 8. - P. 876-893].

Standard for the treatment of iron overload, as well as an antidote in acute poisoning with iron compounds is considered to be Desferal®. It is also known as Deferoxamine, Deferoxamine methanesulfonate, Desferal, Desferal�, Gasperin, DFOM, DFO. On chemical structure is N-[5-{3-[(5-aminopentyl)oxycarbonyl]-propionamido}pentyl]-3-{[5-(N-ACET-amido)pentyl]carbamoyl}-propionyloxy acid. Desferal belongs to the pharmacological group "Detoxifying, including antidotes", CAS code 138-14-7, ATX: V03AC01 (owner of the brand - Novartis Pharma AG, Switzerland) [Desferal:[instruction]./ [Electronic resource]. - Access mode: http://www.likar.info/lekarstva/Desferal/#1]. He has a short elimination half-life from plasma (about 15 min) due to the combination of weak distribution and fast clearance; therefore, the distribution is limited to the plasma and cannot maintain the tank in other cell compartments. The drug is not effective when taken by mouth, absorbed as only 15% of the administered dose.

It forms complexes with trivalent ions of iron and aluminum; less binds divalent ions. This natural siderophore, chelating iron from the labile fraction of the intracellular pool, which quickly updated from the pool of Deposit. Desferal can bind iron, which is in the free form or is part of ferritin and hemosiderin, the Resulting compounds are excreted in the urine, which reduces abnormal deposits of iron in the tissues [Mechanism of action of iron chelators / V. I. Smirnov, G. N. Koltsova, Shcherbinina S. P. [and others] // Hematology and Transfusiology. - 2006. - No. 1. - S. 40-44].

At present, the use of Desferal (Deferoxamine) is common in treatment of patients with iron overload resulting from repeated transfusions. [Ware N. M. Evaluation and treatment of transfusional iron overload in children / H. M. Ware, J. L. Kwiatkowski // Pediatr. Clin. North. Am. - 2013. - Vol. 60, No. 6. - P. 1393-1406]. Minimum absorption of this drug from the gastrointestinal tract and a short half-life in the blood require a slow prolonged parenteral administration of the drug to achieve a negative iron balance as the main goal of effective chelation. Desferal administered in the form of a subcutaneous infusion duration 8-10 hours 3-5 times a week, using a special injector (pump) battery powered [Intensification of chelating-therapy in patients with thalassemia major / J. H. Laws, U. Göbel, A. Christaras [et al.] // Klin. Padiatr. - 2005. - Vol. 217, No. 3. - P. 120-125]. For a long period (almost a lifetime) use of Desferal (Deferoxamine) patients with thalassemia is associated with long-term survival without the complications associated with iron overload [Effectiveness and safety of ICL670 in iron-loaded patients with thalassaemia: a randomised, double-blind, placebo-controlled, dose-escalation trial / E. Nisbet-Brown, N. F. Olivieri, P. J. Giardina [et al.] // Lancet. - 2003. - Vol. 361, No. 9369. - P. 1597-1602]. Unfortunately, around the world, this therapy is assigned only a small number of patients with iron overload, due to poor receptivity to treatment and high treatment costs [yagudina R. I. Pharmacoeconomic analysis chelation therapy of patients with chronic posttransfusion the iron overload / yagudina R. I., Kulikov A. Yu., I. S. Krysanov // Pharmacoeconomics. - 2009. - No. 3. - S. 34-38]. In this case, patients with β-thalassemia, poorly compliant with instructions for the use of the drug, the probability of 25-year survival rate is only 32% [Survival and complications in patients with thalassemia major treated with transfusion and deferoxamine / C. Borgna-Pignatti, S. Rugolotto, P. De Stefano [et al.] // Haematologica. - 2004. - Vol. 89, No. 10. - P. 1187-1193].

The cost and inconvenience of applying Desferal has led to the search for orally active iron chelator, Deferiprone has been used in recent years for the oral treatment of patients suffering from thalassemia [Monitoring long-term efficacy of iron chelation therapy by deferiprone and desferoxamine in patients with beta-thalassaemia major: application of SQUID biomagnetic liver susceptometry / R. Fischer, F. Longo, P. Nielsen [et al.] // Br. J. Haematol. - 2003. - Vol. 121, No. 6. - P. 938-948 Chelation therapy with oral solution of deferiprone in transfusional iron-overloaded children with hemoglobinopathies / A. Makis, N. Chaliasos, S. Alfantaki [et al.] // Anemia. - 2013. - Vol. 2013. - P. 121762]. Deferiprone (Ferriprox) was first approved for use in thalassemia in 1994 and registered in Europe and Asia [Savulescu J. Thalassaemia major: the murky story of deferiprone / J. Savulescu // BMJ. - 2004. - Vol.328, No. 7436. - P. 358-359], much later in the USA and Canada. On chemical structure it is a 3-hydroxy-1,2-dimethylpyridin-4(1H)-one, CAS code 30652-11-0, PBX V03AC02 (owner of the brand - ApoPharma Inc. (Toronto, Canada) [Deferiprone. - Wikipedia the Free Encyclopedia [Electroic resource]. - Regimen of access: http://en.wikipedia.org/wiki/Deferiprone].

The preparation is applied in the iron overload, including myocardial damage [Pat. IL153733 A, IPC A61K 31/44; A61K 31/4412; A61P 9/00. Use of deferiprone for the preparation of iron chelating agents / M. Spino (CA); A. Piga (CA); applicant APOTEX Inc. (CA). - No. IL20020153733; CA20002313270; applic. 29.12.2002; public. 29.04.2010]. He is more comfortable with long-term use, but the risk of developing agranulocytosis, thrombocytopathies and arthropathy limits the use of this drug [Use of deferiprone for iron chelation in patients with transfusion-dependent thalassaemia / S. S. Jamuar, A. H. Lai, A. M. Tan [et al.] // J. Paediatr. Child. Health. - 2011. - Vol.47, No. 11. - P. 812-817; Distal ulnar changes in children with thalassemia and deferiprone related arthropathy / R. Sharma, R. Anand, J. Chandra [et al.] // Pediatr. Blood Cancer. - 2013. - Vol.60, No. 12. - P. 1957-1962].

Another drug for the treatment of iron overload with by oral administration is Deferasirox (Exjade, ICL670, 4-[3,5-bis(2-hydroxyphenyl)-[1,2,4]-triazole-1-yl]benzoic acid) is a chelating drug that is triple-ligand with high affinity to iron (III) and linking it to the 2:1 ratio. Prolonged daily use of Deferaziroksa in adults and children with β-thalassemia or sickle cell anemia on the background of the ongoing transfusions resulted in a reduction of the total body iron, and reduced iron content in the liver was similar to that of Deferoxamine [Deferasirox for managing transfusional iron overload in people with sickle cell diease / J. J. Meerpohl, G. Antes, G. Rücker [et al.] // Cochrane Database Syst. Rev. - 2010. - Vol.8. - CD007477; Long-term safety and efficacy of deferasirox (Exjade) for up to 5 years in transfusional iron-overloaded patients with sickle cell disease / E. Vichinsky, F. Bernaudin, G. L. Forni [et al.] // Br. J. Haematol. - 2011. - Vol.154, No. 3. - P. 387-397].

Deferasirox, as with other iron chelators, may cause side effects (impaired function of the gastrointestinal tract, skin rashes, increased activity of hepatic transaminase and creatinine, high frequency hearing loss, maculopathy and clouding of the lens of the eye (early cataracts) [A maculopapular-type eruption associated with deferasirox administration / A. Ohshita, N. Nakai, N. Katoh [et al.] // J. Am. Acad. Dermatol. - 2013. - Vol.69, No. 5. e265 - 7; P. Chaudhary Deferasirox: appraisal of safety and efficacy in long-term therapy / P. Chaudhary, Pullarkat V. // J. Med Blood. - 2013. - Vol. 4. - P. 101-110].

Deferiprone and Deferasirox - chelators non-biological origin, which is taken orally, some patients who cannot receive Desferal, but these compounds are less effective in binding iron (III) and less toxic. [Comparative efficacy and safety of deferoxamine, deferiprone and deferasirox on severe thalassemia: a meta-analysis of 16 randomized controlled trials / S. Xia, W. Zhang, L. Huang [et al.] // PLoS One. - 2013. - Vol. 8, No. 12. - e82662].

As you can see, the available iron chelators have some problems that require new methods of chelation of iron and the corresponding pharmacological agents, user-friendly, low-toxic and cheap enough, because in most cases it ID�t o long-term therapy.

Describes the process of obtaining chelated iron compounds involving amino-formyl-glycine dipeptide, but the resulting complex compound of iron intended for use in animal husbandry as feed additive, complementary deficiency of minerals, and may not apply in medicine, and its production method is carried out outside the body [Pat. CN 103224546 A, IPC A23K 1/16; C07K 1/113; C07K 5/062. Hydration amino formyl glycine dipeptide chelating iron and preparation method thereof / Fan Mingzhi (CN); Wu Chunli (CN); Xugang Shu (CN); Wu, Xin (CN); Yin Yulong (CN); Gao Junyong (CN); applicant Guangzhou Tanke Bio Technology Co Ltd; Guangzhou Jiuyi Biolog Technology Co Ltd; Zhongkai University of Agriculture and Engineering; Inst, of Subtropical Agriculture Chinese Academy of Sciences (CN). - No. CN 20131142525; applic. 06.05.2013; public. 31.07.2013].

Known to chelate iron compounds and, in particular chelate bisglycinate iron [us Pat. 2480222 OF THE RUSSIAN FEDERATION, IPC A61K 33/26; A61K 31/194; A61K 47/20; A61K 47/26; A61P 7/06. Pharmaceutical preparations containing chelate bisglycinate iron / M. Balducci (IT); stated. and the patentee M. Balducci (IT). - No. 2009104689/15; Appl. 11.02.2009; publ. 27.04.2013]. In this case, the receiving chelating aimed to use as the source of iron deficiency in the body and may not apply in the case of acute and chronic iron overload.

There is a way complexation to obtain the compounds of metals, specifically iron chelate in the form of its concentrated solution to�th is carried out by interaction of iron salts in an aqueous medium with a complexing agent, which is used as N,N,N',N'-ethylenediaminetetraacetic acid and citric acid, [Pat. 2458930 OF THE RUSSIAN FEDERATION, IPC C07F 15/02. The method of obtaining humic-containing chelate iron / V. A. Brykalov (RU), E. M. Golovkin (RU), E. V. long (RU); stated. and patent holder of HPE "Kuban state agrarian University" (RU). - No. 2010150895/04; Appl. 13.12.2010; publ. 20.08.2012]. This method of iron chelation is simple, but takes place outside of the body and aims to obtain a solution for the nutrition of agricultural plants. It cannot be used for iron chelation as a therapeutic intervention in pathological conditions associated with overload of the body this element.

Proposed binding of iron ions using water-insoluble polymeric chelating agent, a polymer having the main chain and an aromatic ring attached to the chain through relationship-NH-CH2where the aromatic ring has the functional group in the form of-OH, -COOH or other radicals [Pat. ER A1, IPC A61K 31/722; A61P 13/12; A61P 3/12; A61P 39/04. Polymeric iron chelating agent / Yuzo Nishida (JP); Yutaka Kohgo (JP); Katsuya Ikuta (JP); Katsunori Sasaki (JP); string Disease Adsorption System Technologies Co Ltd (JP); Nat. University Corp. Asahikawa Medical University (JP). - No. EP 20120734667; JP 20110006043; applic. 12.01.2012; public. 20.11.2013]. The insolubility in water suggests the possibility of selectively chelating biologically unstable iron, not joining in share�record processes in vivo however, such a chelator is not useful for medical applications, especially for sterile dosage forms required for parenteral administration in case of acute poisoning by the salts of iron.

Known iron chelator that is used together with pyrithione [Pat. MH A, IPC A61K 31/555. Personal care compositions comprising a pyrithione and an iron chelator / C. W. Saunders; Youngquist, R. S.; Jun, X.; Kelly, C. P.; Domsic, J. K.; Lucas, R. L.; Schwartz, J. R. (US); applicant Procter&Gamble (US). No MX 20130004557; US 20100407754P; applic. 23.04.2013; public. 07.06.2013], but this composition is intended for hygienic purposes and is not suitable for the treatment of acute or chronic overload of iron in the body, and also for elimination of iron ions in free radical pathology.

As iron chelators available Ecthelion, but the emphasis in this invention is done on the antioxidant properties of these substances and their introduction in myocardial infarction with reperfusion of the myocardium to prevent damage to the heart muscle as a result mediated by iron ions free radicals formation [Pat. MH A, IPC A01N 1/02; A61K 31/55. Novel iron chelator and inhibitor of iron-also been other ideas where oxidation / Lawrence Horwitz; Marcus Horwitz, A.; Gibson, Bradford W.; Joseph Reeve (US); applicant Univ. California (US). No MX 19960004499; US 19950383180; applic. 01.10.1996; public. 29.11.1997].

The chelation of iron is proposed with phytic acid or its salts and lactoferrin, which have complexing�obrazuemymi properties and at the same time, as nutrients, have low toxicity, however, mentioned the composition is focused only on the improvement of the skin and the use in cosmetic or dermatological preparation [Pat. DE102005019202 A1, IPC A61K 8/18; A61K 8/55; A61K 8/67; A61K 8/98; A61Q 17/00; A61Q 19/00. Cosmetic/he is considered to be preparation, useful e.g. to improve the skin appearance, comprises iron chelating agent such as phytin and/or its slats or lactoferrin / K. Sauermann (DE); Sauermann C. (DE); G. Sauermann (DE); Jaspers S. (DE); U. Koop (DE); Filbry A. (DE); applicant Beiersdorf AG (DE). - No. DE 20051019202; applic. 20.04.2005; public. 26.10.2006].

Proposed complexing agents of General formula, which may be a carboxyl group or its derivative, hydroxyl, gidrauxilirovannaya alkoxy - or alkoxyalkyl group; methyl group and the nitrogen atoms and sulfur used to obtain diagnostic and pharmaceutical products for nuclear magnetic resonance imaging, scintigraphy, ultrasound, radiotherapy, as well as for detoxification in cases of poisoning by compounds of heavy mellow [us Pat. 2073005 OF THE RUSSIAN FEDERATION, IPC C07F 11/00, C07F 15/02. Chelate compound of the metal / Berg (NO), Alma (SE), Th. Klaveness (SE), P. Rangvid (SE), Thomassen (NO); stated. and patent holder Nycomed AC (NO). - No. 4743079/04; Appl. 15.07.1988; publ. 10.02.1997]. However, the proposed chelators are not specifically recommended for the treatment of an overload of iron in the body, or acute intoxication with salts of iron. In addition, based on the chemical �re proposed complexions, we can assume that they have significant toxicity, particularly hepatotoxicity.

Known synthesis and description of derivatives of pyridoxal isonicotinohydrazide hydrazone suitable for oral dosage form, soluble in water and are capable of chelation in conditions of iron overload and other metals [Pat. US5834492 A, IPC C07D 213/86. Water soluble orally effective iron chelator / Lewis N. (US); Patel V. (US); Terpinski J. (US); R. Bliss (US); applicant Jacobus Pharmaceutical Co (US). - No. US19970765369; applic. 07.04.1997; public. 10.11.1998], but we are talking about substances that are in the preclinical stage of testing and can have a significant amount of side effects due to the structural analogy with vitamins and antagonism with them.

There is an invention that relates to chelating agents with high stability and high chelating ability to iron, surpassing the activity of ethylenediaminetetraacetic acid [Pat. CN 103304435 A, IPC SS 227/10; SS 229/36. Chelating agent with high stability and high iron chelating ability and preparation method thereof / Xue Qunxiang (CN); Gong Feixiang (CN); applicant Shaanxi Res. Design Inst, of Petroleum and Chemical Industry (CN). - No. CN 20131277722; applic. 03.07.2013; public. 18.09.2013]. A chelating agent is a compound, which is obtained from phenol derivatives, aliphatic diamine and an aqueous solution of acid glyoxalic in one-step synthesis, however, the prospects of clinical application are described�wow chelator remain unclear.

Known invention relating to a chelating agent that can selectively chelate iron ions and is represented by the compound or its salt containing an aromatic hydrocarbon ring or aromatic heterocyclic ring radicals in the form alkilinity groups, hydrogen atoms, hydrocarbon group or a group having chelating ability [Pat. US 2011189779 A1, IPC SS 229/14; SS 229/26; C07D 233/64; G01N 33/20. Iron chelating agent, method for producing same, method for determining amount of iron ions and method for trapping iron ions / Kohg Yutaka (JP); Katsuya Ikuta (JP); Katsunori Sasaki (JP); Yuzo Nishida (JP); applicant Univ. Yamagata (JP); Nat. Univ. Corp. Asahikawa Medical College (JP). - No. US 200913120126; JP 20080243095; applic. 18.09.2009; public. 04.08.2011], but, as in the previous case, the possibility of clinical use of this chelating agent has not been studied.

There is an invention relating to cycloalkenes derived 3-hydroxy-4-pyridinone, which is useful for complexation with metal ions, including iron, in chemical and biological systems [Pat. NZ529657 A, IPC A61K 31/4422; C07D 213/81. Iron chelating cycloalkyl derivatives of 3-hydroxy-4-pyridinones / Tam T.; Spino M.; Li, W.; Wang Y.; Hao Y.; Shah, B. H.; applicant APOTEX Inc. (CA). - No. NZ20030529657; applic. 20.11.2003; public. 30.07.2004], however it is possible to believe that such a derivative, as well as other known substances of this series, will have side effects that limit its possible use in patients with acute and chronic iron overload.

Hal�aims to iron can be carried out with the use of the derived pyrazolone, containing aromatic radicals and halogen atoms, which, obviously, along with the elimination of excess iron in the body of animals and humans, will cause the voltage of the detoxification system in the liver [Pat. JP 2004203820 A, IPC A61K 31/4152; A61P 3/10; A61P 43/00; A61P 9/10; C07D 231/26. Iron chelating agent / Tamaoki Toshiaki (JP); Koichiro Tsuchiya (JP); applicant Mitsubishi Pharma Corp. (JP). - No. JP20020376668; applic. 26.12.2002; public. 22.07.2004].

Proposed new crystalline forms of 4-[3,5-bis(2-hydroxyphenyl)-[1,2,4]triazole-1-yl] benzoic acid; methods of obtaining these crystalline forms, compositions containing these forms, and their use in diagnostic methods or treatment of warm-blooded organisms, especially humans [Pat. US 2012203007 A1, IPC C07D 249/08. Polymorphic forms of deferasirox (ICL670A) / M. Mutz (DE); applicant Novartis AG (CH). - No. US 201213405428; EP 20060125002; WO 2007EP62903; applic. 27.02.2012; public. 09.08.2012]. However, in the present invention, refers to already used the tool Deferasirox and a new form or pharmaceutical composition on its basis and the method of application for the chelation of iron can drastically reduce the side effects of this drug.

The chelation of iron overload of iron in the body are analogues of desperation and describes their use [Pat. US 2013210870 A1, IPC C07D 277/12. Desferrithiocin Polyether Analogues / Bergeron Jr. Raymond J. (US); applicant Univ. Florida (US). - No. US 201213683301; WO 2008 US 03433; applic. 21.11.2012; public. 15.08.2013]. However, the claimed to invent the�research Institute of matter characterized nephrotoxicity in animal experiments. In addition, the proposed compounds are in the preclinical stage of the study and it is impossible to say what will be the results of clinical trials, because the data disclosed a significant species specificity.

Known invention relating to the amphiphilic chelators of metals that have the specificity to iron and demonstrate neuroprotective and good transport properties in lipophilic environments [Pat. US 2012058945 A1, IPC A61K 31/4412; A61K 31/47; A61K 31/497; A61K 38/00; A61K 38/09; A61K 38/22; A61K 45/06; A61P 25/00; A61P 25/16; A61P 25/28; C07D 213/69; C07D 215/26; C07D 295/15; C07D 295/205; C07K 14/575; C07K 14/70; C07K 5/10; C07K 7/06; C07K 7/22; C07K 7/23. Neuroprotective iron chelators and pharmaceutical compositions comprising them / M. Youdim (IL); M. Fridkin (IL); H. Zheng (IL); A. Warshawsky (IL); R. Warshawsky (IL); applicant Yeda RES & DEV (IL); Technion RES &DEV Foundation (IL). - No. US 201113296061; WO 2003IL00932; applic. 14.11.2011; public. 08.03.2012]. Function chelator of iron is provided preferably residues of 8-hydroxyquinoline, hydroxamate or pyridinone. Compounds of the present invention are useful for the treatment and prevention of diseases, disorders or conditions associated with iron overload and oxidative stress, namely neurodegenerative and cerebrovascular diseases, tumor diseases, hemochromatosis, thalassemia, cardiovascular disease, diabetes and several other diseases. The proposed N-hydroxypyridine-2-ones and 3-hydroxypyridine-4-ones perspective� for oral use (hydroxypyridinone CP20, or Deferiprone is used in clinical practice); some hydroxypyridinone (for example, CP20, CP24, CP94) able to penetrate the blood-brain barrier and have structural similarity between the catechol group is a chelator of iron in the structure of DOPA. In another embodiment of this method the function of chelation is provided by hydroxamates, as hydroxamate known as iron chelators, and desferrioxamine In (Desferal) has long been the drug of choice for treatment of iron overload. As you can see, the main emphasis in this invention is the use of previously known iron chelators for new indications and suggests ways of using new agents for the chelation of iron.

Closest to the claimed (prototype) is a means to reduce the content of metals, especially iron, in mammals, representing conjugates of Deferoxamine, derivatives and analogues [Pat. US 2010273847 A1, IPC A61K 31/16; A61K 31/4188; A61P 1/16; A61P 25/00; A61P 31/10; A61P 33/06; A61P 39/04; C07C 259/06; C07D 495/04. Desferrioxamine conjugates, derivatives and analogues / R. Codd (AU); G. L. Schipanski (AU); applicant R. Codd (AU); G. L. Schipanski (AU). - US 20080740957; AU 20070905998; WO 2008AU01617; applic. 31.10.2008; public. 28.10.2010.]. The compounds referred to in this way (Biotin-deferoxamine, adamantane-deferoxamine, benzo-15-crown-5-4-carboxyl-deferoxamine or 4-methyl-phenoxyacetate-deferoxamine), like himself Deveraux�mine are iron chelators and can be used in case of overload of iron in the body as a result of dysmetabolic disorders and regular blood transfusions. They can also be used to treat cancer, malaria, fungal infections, the development of which the role of iron and (or) free-radical oxidation. Such compounds have a greater hydrophobicity compared with deferoxamine and the possibility of oral administration, they are more resistant to enzymatic degradation and have a longer half-life. Believe that the presence of functional groups in the N-terminal sequence of new connections will reduce the side effects associated with this section of the Deferoxamine molecule. In addition, conjugation with substances such Deferiprone or Desferasirox makes it possible to participate in the chelation of iron at both ends of the molecule. At the same time, the proposed analogues, insufficiently studied in experiments on animals, especially not studied and have not been applied in the clinic, which does not allow sufficiently to evaluate their effectiveness and safety.

Thus, the disadvantages of the known means of iron chelation is that complexing agents are substances that have many side effects or pharmacokinetic properties that hinder their clinical application (e.g., short half-life, limited distribution, or the need to use only one route of administration), and n�emerging at the stage of preclinical trials with uncertain prospects of use in the clinic.

The object of the invention is to expand the Arsenal of tools the chelation of iron ions, which ensure the reduction of its content in the body when enteral and parenteralna the introduction of a minimal toxicity, as proven in Toxicological and clinical studies.

The problem is solved by using as a chelating means 2-ethyl-6-methyl-3-hydroxypyridine succinate (Mexidol) for the treatment of an overload of iron in the body or hyperchromatosis.

The technical result is that when using this agent in acute and chronic load iron sufficiently high efficiency of the process of chelation with low toxicity Mexidol, which provides the prospects of its biomedical applications.

Mexidol belongs to a known class of compounds, which includes a number of substances exhibiting biological properties [Djumaev K. M. Structure and reactivity of derivatives of 3-hydroxypyridine in electrophilic substitution / K. M. Dunaev, L. D. Smirnov // Uspekhi khimii. - 1975. - Vol. 44, No. 10. - S. 1788-1804]. This class of compounds has antioxidant properties and is widely used in medicine [Pharmacology of antioxidants on the basis of 3-hydroxypyridine / V. E. Novikov, L. A. Kovaleva, S. O. Lisenkova [et al.] / / access Mode to article: ].

Mex�Dol - a substance with the empirical formula C12H17NO5and molar mass 255,26 g/mol, very soluble in water and ethanol, slightly soluble in ether and insoluble in chloroform, with a maximum absorption at a wavelength of 297±2 nm in the identification method of UV spectroscopy [etilmetilgidroksipiridina succinate. Substance powder. Manufacturer: LLC "Bion". Per. number: LSR-001704/07-260707].

It belongs to the group ATX N07XX, CAS 2364-75-2; produced in such dosage forms as capsules, solution for intravenous and intramuscular injection, coated tablets, tablets, film-coated under the trade names Mexidol®, Megamaxi, Maximal, Oxidant, Mexicor, Moexipril, Maximin [etilmetilgidroksipiridina succinate. Wikipedia, the free encyclopedia [Electronic resource]. - Access mode:

The brand owner Mexidol® - NPK "pharmasoft" (Russian Federation).

2-ethyl-6-methyl-3-hydroxypyridine succinate used in the clinic and has demonstrated antioxidant, antihypoxic, anxiolytic, nootropic effect, improves myocardial function and liver, has radioprotective properties [Voronina T. A. Antioxidant Mexidol. Main neuropsychotropic effects and mechanism of action / T. A. Voronina // Psychopharmacology and biological narcology. - 2001.- No. 1. - C. 2-12; Mexidol and concomitant vascular pathology of the brain and heart / R. S. Mirzoyan, T. S. Genunine, M. A. Lebedev [et al.] / / Experim. and wedge, pharmacology. - 2011. - Vol. 74, No. 6. - Pp. 20-23; the Effect of Mexidol on postirradiation system restore hematopoiesis / V. V. Moroz, Y. B. Cheap, G. W. Scoan [et al.] / / well defined spatial radiation viewing. Biol. radio-Ecolog. - 2009. - Vol. 49, No. 1. - S. 91-96].

Lethal dose of 2-ethyl-6-methyl-3-hydroxypyridine succinate which causes death in 50% of animals (LD50), is equal to rat 820 mg/kg for mice and 475 mg/kg, and when administered orally over 3000 mg/kg in rats and 2010 mg/kg in mice. Comparison of effective therapeutic doses (ED) of Mexidol (10-300 mg/kg) toxic and lethal doses shows significant therapeutic breadth and security tools: therapeutic index, calculated as the ratio LD50/ED50 is 16.4 [Voronina T. A. MEXIDOL® main effects, mechanism of action, use [Electronic resource] / T. A. Voronina. - Access mode:

Specified derivative 3-hydroxypyridine has favorable pharmacokinetic properties. It is rapidly absorbed when taken orally (partly suction period - 0,08-1 hour). Intramuscular injection is determined in plasma for 4 hours after injection with achievement of maximal concentration in intramuscular using 0.3-0.58 hours when taken in�UTR - 0,46-0.5 hours. Rapidly distributed to organs and tissues. Average retention time of drug in the body after intramuscular injection of 0.7-1.3 hours, the ingestion of 4.9 to 5.2 hours. Metabolized in the liver by glucuronidation. Rapidly excreted in the urine mainly as metabolites (50% for 12 hours). [Pharmacokinetic, behavioral, and neurophysiological aspects of the action of 2-ethyl-6-methyl-3-hydroxypyridine in rats / A. K. Sariev, S. V. Krapivin, T. A. Voronina [et al.] / / bull. experim. of biology and medicine. - 1998. - Vol. 106, No. 8. - Pp. 165-167; the Relationship of glucono-nonconjugate Mexidol and features of its therapeutic action in patients with organic lesions of the CNS / A. K. Sariev, I. A. Davydov, G. G. Neznamov [et al.] / / Experim. and wedge, pharmacology - 2001. - Vol. 64, No. 3. - S. 17-21].

To obtain the effect of chelation in vivo, a solution of 2-ethyl-6-methyl-3-hydroxypyridine succinate containing an effective dose of the drug (100-50 mg/kg of body weight in animal experiments), administered at the same time orally or parenterally in the acute load iron or multiply one of these ways to achieve negative iron balance in chronic situations.

As a criterion for the evaluation of the connection formation between 2-ethyl-6-methyl-3-hydroxypyridine the succinate and iron ions in the model system chosen electronic scanning microscopy and interfactory and�Aliz, to determine the elemental composition of the compounds formed and their compliance with the previously known substances, and in the body - determination of iron concentration in serum and iron-sequestering organs (liver, spleen).

The invention is illustrated by the following examples.

Example 1. The interaction of blood plasma with ions of iron (III) outside the body

Materials and methods. The crystal hydrate of ferric chloride (III) (FeCl3) was dissolved in distilled water in an amount necessary to prepare a 1% solution (weight/volume). The concentration of ions of Fe3+before and after interaction with 2-ethyl-6-methyl-3-hydroxypyridine the succinate controlled photocolorimetry with the use of salicylic acid or potassium thiocyanate on the previously prepared calibration curve. Substance 2-ethyl-6-methyl-3-hydroxypyridine succinate was dissolved in 1% solution of ferric chloride (III) in an amount such that their molar ratio was 2:1. The obtained reaction product were examined for solubility in polar and nonpolar solvents and a melting temperature. Selected crystals were subjected to washing on the filter SCHOTT hot solvents in the following sequence: water-DMFA-N-methylpyrrolidone-toluene-ethanol-hexane, and then dried at 90-100°C. the Purified crystals were subjected to elemental analysis�. For the quantitative determination of carbon and hydrogen was used the method of Prepa for the quantitative determination of nitrogen - Dumas method [lip veil. Methods of organic chemistry: Methods of analysis. Vol. 2. - M.: Goskomstat, 1963. - S. 106-109, 180-188]. In addition, the obtained complex was investigated by scanning electron microscopy with spectroscopy using a raster scanning electron microscope (Cam Scan 4D (CamScan Electron Optics Ltd, UK) [Gorelik S. S. Radiographic and electronoptical analysis: Proc. manual for schools: 3rd ed. EXT. and Rev. / by S. S. Gorelik, L. N. Rastorguev, Yu. A. Skakov. Moscow: MISIS, 1994. - 328]. On the basis of elemental analysis calculated the empirical formula of the resulting complex.

Implemented roentgenoscopy sample analysis [Gorelik S. S. Radiographic and electronoptical analysis: Proc. manual for schools: 3rd ed. EXT. and Rev./ by S. S. Gorelik, L. N. Rastorguev, Yu. A. Skakov. Moscow: MISIS, 1994. - 328 p.]. The diffraction patterns were recorded on a diffractometer DRON-UM1 (OAO NPP "Burevestnik", Russia) at the emission of Co Ka line anode (1.79021 Å) with a graphite monochromator in the reflected beam when shooting geometry for Bragg-Brentano (in angular range 4-80 degrees with a step of 0.1 deg and exposure of 1 s. For the study of thermal degradation of the reaction product it is sequentially heated in air to temperatures of 120, 200, 300 and 400°C and record�Ali cooled diffraction pattern of the samples. According to interfaceware analysis attempted to identify the reaction product in the database PDF-2 [PDF-2. PCPDFWIN Versio129619n 2.1, - 2000, JCPDS-ICDD].

Results and discussion. In the interaction of 2-ethyl-6-methyl-3-hydroxypyridine succinate with 1% solution of FeCl3in the ratio 2:1 forms a gel-like substance structure, dark red. The supernatant is qualitative reaction for ions of Fe+3with potassium thiocyanate (KSCN), showed their absence, indicating that the iron had reacted completely.

At low concentrations of FeCl3the gel is not formed. When the concentration of Fe+3the substance acquires a deep red color and greater density. When drying of the gel formed white crystals and amorphous substance of a red color.

White crystals soluble in water, but more soluble in ethanol. Upon evaporation of the solvent they have the same melting point as 2-ethyl-6-methyl-3-hydroxypyridine succinate. Sample mixing does not detect depression of the melting temperature, which suggests that this is 2-ethyl-6-methyl-3-hydroxypyridine succinate, which is not reacted.

Since the unreacted substance is well soluble in ethanol, purification of the complex was performed using the solvent, until then, until the supernatant is not re�taval be detected 2-ethyl-6-methyl-3-hydroxypyridine succinate (white needle crystals, identified by melting point) followed by rinsing with hot organic solvents on the filter SCHOTT.

Exploring the resulting reaction product by comparison of melting points, it was shown that this material is 2-ethyl-6-methyl-3-hydroxypyridine the succinate, which has a melting point of 112°C. To the obtained compound, the melting point is 128 to 130°C, which shows that it is not only different from Mexidol, but also more complex molecular structure.

It was also found that in the interaction of the gel formed in the reaction between FeCl3and derivatives 3-hydroxypyridine, with acid chloride (1:1) it dissolves; staining disappears. The proton acid chloride may accede to the nitrogen atom of the pyridine ring, which contains an unshared pair of electrons. This leads to disruption of the gel structure, which indirectly confirms the involvement of nitrogen atom in its formation.

The results of the study of the complex by elemental analysis showed that it contains: C - 25,63%; 25,74%; H- 4,05%; 4,22%; 3,95%; N - 1,01%; 1,04%.

When studying the reaction product of Fe+3and derivatives 3-hydroxypyridine by scanning electron microscopy with mass spectrometry is shown in its crystalline structure (Fig.1) and four changes�relations in stochastically selected sites installed content of elements in mass %: C to 33.52-35,29%; O 46,12-60,95%; Fe 4,02-18,85%. The content of elements in atom%: C 41,98-46,10%o; O 47,62-56,52%; Fe from 1.07 to 5.58%. The discrepancy between the results of the analysis, especially on the carbon content may be due to the peculiarities of the methods used, in particular, use a hardware-based carbon-containing tape during electron microscopy.

The diffraction pattern of the initial sample substances produced in the reaction between Fe+3and 2-ethyl-6-methyl-3-hydroxypyridine the succinate, are shown in Fig. 2. It is characterized by the presence of several broadened and partially overlapping diffraction peaks, indicating a weak (low) crystallinity of the material tested. Characteristics of the peaks are presented in Table 1.

In the framework of x-ray diffraction database PDF-2 the substance of the reaction product are not identified. The presence of iron in the composition of the sample confirmed the formation of iron oxide (II, III) Fe3O4when heating the sample in air to 300°C, which is manifested in the change of the diffraction pattern to the characteristic of the oxide (Fig. 3, curve 4).

Comparison of the diffraction pattern of the obtained complex diffraction pattern with 2-ethyl-6-methyl-3-hydroxypyridine succinate [us Pat. 2453538 C1 of the Russian Federation, IPC C07D 213/65. Stable crystalline form of 2-ethyl-6-methyl-3-hydroxypyridine bitches�of hinata and method of its production / Herman SAT. (RU), Gamin A. M. (RU), O. L. Germanova (RU), Filippova E. S (EN), Missoul A. B. (RU); stated. and patent holder LLC "PharmaMed" (RU). - No. 2011114873/04; Appl. 06.04.2011; publ. 20.06.2012.] detects a significant difference in the position and intensity of their peaks, which may indicate that the analyzed substance, different from the original substances, which entered into reaction.

Sample heating up to 120 and 200°C leads to a directed change in the intensity of the diffraction peaks is a decrease in the intensity of peaks No. 1, 3, 5, and increase the intensity of the peak No. 2 and, thus, to the partial destruction of its structure (see Fig. 3). A further temperature increase causes the disappearance of the peaks characteristic of the starting sample, and the appearance of peaks of iron oxide (II, III), indicating a destruction of the organic component of the material tested. Data on thermal transformations also favor the formation of a new organo-inorganic compounds based on the source component.

The formation of coloured complex in the reaction between iron and 3-hydroxypyridine can be explained by the fact that 2-ethyl-6-mthyl-3-hydroxypyridine succinate is in the phenolic nucleus, which is the typical reaction of interaction with FeCl3leading to the formation of the complex, shown in Fig. 4.

Calculated according to the element�wow analysis the simplest formula of the product (or its kristallogidrata) the reaction between 2-ethyl-6-methyl-3-hydroxypyridine a succinate and a soluble salt of iron (III) has a view of FeC 11-15H21N3O18.that suggests that the binding of 1 mol of iron ions involved 3 mol derived 3-hydroxypyridine in partial degradation of its molecules. At the same time, the comparison of the empirical formula with the formula 2-ethyl-6-methyl-3-hydroxypyridine succinate suggests that the actual ratio of atoms in a molecule is a multiple of a basic, and are present in the molecule coordination. Because the substance formed in the reaction between 2-ethyl-6-methyl-3-hydroxypyridine the succinate and Fe+3has the properties of a gel, it is also likely to exist in the structure of hydrogen bonds.

Thus, as a result of chemical interaction of 2-ethyl-6-methyl-3-hydroxypyridine succinate ions with Fe3+formed persistent colored complex with high molecular weight and complex component (organic-inorganic) composition, not identified among the previously known compounds.

Example 2. The action of Mexidol in the acute load of iron in the body

Materials and methods. In the experiment used white rats of Wistar line weighing 180-200 g (PE "Biomolecules", Ukraine). During the experiment they were kept in standard plastic cages and received pelleted feed for laboratory rodents (PE "Biomolecules",Ukraine) and water demand. The experiments received approval by the Commission of bioethics of UGUSU "Ukrainian medical stomatological Academy".

The grouping of the experiments had the following appearance at 6-7 animals in each group:

1. Intact rats;

2. Intact rats with the introduction of the solvent (control);

3. Acute load of iron (reference pathology);

4. Acute load iron + Mexidol;

5. Acute load iron + Desferal.

As the load of the substance used iron (III) chloride FeCl3in toxic dose of 2000 mg/kg body weight, was administered intragastrically once with a probe in the form of a 20% solution of FeCl3prepared from 6-water kristallogidrata. When choosing the dose was guided by the literature data that the administration to rats per os anhydrous FeCl3LD50=450 mg/kg [Ferrous chloride, 4-hydrate. - MSDS Number: F1678. Effective date 11/10/05. Supercedes 02/18/03 [Electronic resource]. - Regimen of access: http://2.imimg.com/data2/MM/RV/MY-893306/ferrous-chloride-tetrahydrates.pdf].

Mexidol was administered to the animals 10 minutes after injection of a solution of ferric chloride at the same time intragastrically and intraperitoneally. His solution was prepared ex tempore by using substance 2-ethyl-6-methyl-3-hydroxypyridine succinate (Bion, Russia). The stomach tube was introduced 1 ml of a 25% solution of Mexidol (1250 mg/kg body weight) intramuscularly with 0.5 ml of 4% solution of Mexidol (100 mg/kg mA�si body). The dose was chosen based on the range of effective and toxic doses of this drug [Voronina T. A. MEXIDOL® main effects, mechanism of action, use [Electronic resource] / T. A. Voronina. - Access mode: http://medi.ru/doc/a 070196.htm], and based on the analogy with the use in such a situation, a known chelator of iron - Desferal. Desferal (Novartis Pharma, Switzerland) was used as a comparison drug. His solution was prepared from lyophilized powder in vials ex tempore and was administered intragastrically (250 mg/kg) and intraperitoneally at a dose of 80 mg/kg. The dose and route of administration of Desferal was chosen in accordance with the recommendations for its use in acute poisoning with salts of iron [Desferal: [instruction]. / [Electronic resource]. - Access mode: http://www.vidal.ru/poisk_preparatov/desferal_232.htm]. The rats in the control group thus introduced the solvent is water for injection. After 5 days, animals were killed under ether anesthesia by means of blood collection from the heart until it stops. In the serum, liver and spleen was determined by the iron content by atomic emission spectrometry with inductively coupled plasma instrument Optima 2100 DV (Perkin Elmer, USA) [Determination of chemical elements in biological fluids and drugs by means of atomic emission spectrometry with inductively coupled plasma and mass spectrometry with a water-saving model�UPE connected plasma: 4.1.1482-03. - The official. ed. - M.: The Ministry Of Health Of Russia, 2003. - 16 S. - (Normative document of the Ministry of health of the Russian Federation. Methodical instructions)]. The obtained data were statistically processed using the computer program Statistica for Windows V. 8.0.

Results and discussion. Within 5 days from the date of introduction of iron salts is not marked changes in the behavior and appearance of animals, with the exception of staining of urine in a brown-red color in the groups treated with Mexidol and Desferal. When inspecting the internal organs of animals after euthanasia macroscopic changes characteristic of toxic shock, is not identified.

The iron content in serum, liver and spleen of animals are presented in Table 2.

Indicators of iron metabolism animals with the introduction of the solvent did not differ from the parameters of intact white rats (see Table 2).

Load of iron was characterized by the increase of iron content in serum of animals 3.4 times (p<0,001) compared to the control. The iron content in the liver was increased 1.4-fold (p<0.05) and in the spleen was increased 1.5-fold (p<0.05) compared with control values.

The use of Mexidol on the background load of iron caused a decrease in the concentration of this element in serum 3 times (p<0.002 inch) compared to the control p�Teologia (see Table 2). In animals of this group, the iron content in the spleen was decreased 1.5-fold (p<0.05) compared with the introduction of iron salts without pharmacological correction. In the liver under the influence of Mexidol significant changes in iron content were observed, but there was a trend towards lower concentrations (p<0,1).

When you use Desferal there was a decrease of iron concentration in serum was 2.9 times (p<0.05) compared with the pathological background (see Table 2). The content of iron in the liver of animals decreased in 1.7 times (p<0.01) and was significantly lower not only than with a load of iron without pharmacological treatment, but also in the control groups. At the same time, the content of this element in the spleen did not change significantly and was significantly higher than when therapeutic introduction of Mexidol.

Thus, when a single load of iron (2000 mg/kg, intragastric) Mexidol, introduced intragastrically (1250 mg/kg) and intraperitoneally (100 mg/kg), decreases the content of this element in serum and "load" the authorities that the severity of the blood is not inferior, and in the spleen greater than the effect of the standard iron chelator - Desferal.

Example 3. The action of Mexidol in the chronic load iron

Materials and methods. In the experiment used white wing�s-males of Wistar line weighing 180-200 g (PE "Biomolecules", Ukraine). Before and during the experiment they were kept in standard plastic cages and received pelleted feed for laboratory rodents (PE "Biomolecules", Ukraine) and water demand. The experiments received approval by the Commission of bioethics of UGUSU "Ukrainian medical stomatological Academy".

The grouping of the experiments had the following appearance, with 8 animals in each group:

1. Intact rats;

2. Intact rats with the introduction of the solvent (control);

3. Intact rats with the introduction of solvent + Mexidol;

4. Chronic load of iron (reference pathology);

5. Chronic load iron + Mexidol.

Chronic load iron reproduced by repeated parenteral administration to animals of a preparation of iron [Study of the eye and lacrimal glands in experimental iron overload in rats in vivo / M. Repanti, S. P. Gartaganis, N. M. Nikolakopoulou [et al.] // Anat. Sci. Int. - 2008. - Vol. 83. - P. 11-18]. As such a drug used iron (III) hydroxide of polyisomaltosate in the form of a preparation ferrum Lek (Lek, Slovenia) [FERRUM LEK® for the on/m introduction (FERRUM LEK). Reference Vidal drugs in Russia [Electronic resource]. - Access mode: ~]. It was injected to rats intraperitoneally at a dose of 7.5 mg Fe/1 animal every 3-4 days for 4 weeks, which amounted to 10 injections. One group of animals at the same time inside�rushino Mexidol was administered at a dose of 50 mg/kg of body weight. A solution of Mexidol was prepared ex tempore from its substance, obtained from the manufacturer (Bion, Russia), using as solvent water for injections. Animals of the control groups were injected solvent or a solution of Mexidol (50 mg/kg), following the specified schema. 3 days after the last injection the animals were euthanized. During the autopsy was performed the examination of the internal organs. In the serum, liver and spleen was investigated the content of elemental iron as described in example 2. The obtained data were statistically processed using standard computer programs.

Results and discussion. The content of Fe in serum, liver and spleen of animals that received injections of the solvent (control) were the same as in intact animals (table. 3).

Course introduction of Mexidol in the dose of 50 mg/kg of intact animals caused a tendency to Snejniy iron content in serum and load bodies in comparison with intact animals (p<0.1) and in the spleen and compared with the control group (p<0.2) (see Table 3).

Chronic load of iron was characterized by increased serum iron in 1.4 times (p<0,02) compared with intact animals and 1.6-fold (p<0,005) compared with the control (see Table 3). The content of this element in �Iceni was increased by 1.6 times compared with the intact group (p< 0.05) and 1.9-fold (p<0,001) compared to animals of control group received injections of the solvent. In the spleen of rats with a load of iron was observed with increasing the concentration of iron in 1.3-1.4 times (p<0.05) compared with intact animals and control. Such changes may indicate increased as transport pool of iron and its deposition on the background of the introduction of this additional element for 4 weeks in total dose 375 MrFe/kg of body weight.

Mexidol has reduced the level of serum iron in 1.3 times (p<0.05) compared with load iron without pharmacological treatment (see Table 3). The liver and spleen iron content was decreased equally in 1.4 times (p<0,005) compared to the control pathology.

As you can see, the use of Mexidol (50 mg/kg) during chronic load of iron reduces the accumulation of this element in the body and normalize its content in the blood serum and organs depot.

Thus, the experiments shown that there is a possibility of removal of excess iron from the body in the case of acute or chronic load this element through the use of derivatives of 3-hydroxypyridine, in particular 2-ethyl-6-methyl-3-hydroxypyridine succinate, based on the chemical interaction between this production�, relatives and ions of iron (III) with the formation of persistent colored complex with high molecular weight and complex component (organic-inorganic) composition, unidentifiable among the previously known compounds. The inventive method may have a biomedical application that is provided as its immediate result in the form of elimination of excess iron from the body, and the pharmacological and Toxicological properties of 2-ethyl-6-methyl-3-hydroxypyridine succinate (Mexidol), offered as a chelating agent. Effective dose (100-50 mg/kg of body weight in animal experiments) injected simultaneously orally or parenterally in the acute load iron or multiply one of these ways to achieve negative iron balance in chronic situations.

The use of 2-ethyl-6-methyl-3-hydroxypyridine succinate as chelating agents for treatment of an overload of iron in the body or hemochromatosis.



 

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15 tbl, 5 ex

FIELD: medicine.

SUBSTANCE: melanin having water-solubility of at least 80% and an paramagnetic centre concentration of at least 8·1017 spin/g is administered orally into the animals having been exposed to the radiation in a dose high enough to cause a spinal radiation injury; melanin is administered after dissolved in distilled water in the effective concentration. Melanin water is used as drinking water for the mice having been exposed to single and fractionated acute radiation, which is able to cause acute radiation disease. Melanin water is taken from the 1st to 30th day following the single radiation, or from the 1st day of the fractionated radiation to the 30th day on completion of the radiation.

EFFECT: higher survival rate, faster recovered haemopoiesis, body weight and orientation and motion activity.

7 tbl, 5 ex

FIELD: chemistry.

SUBSTANCE: invention provides a composition having antioxidant properties in the form of a tablet, comprising an active agent based on nicotinamide adenine dinucleotide in reduced form (NADH) and inert filling agents, characterised by that the active ingredient is a complex which is a mixture of 10 wt % NADH with 63 wt % vegetable fats, 17 wt % beeswax and 10 wt % chlorophyll, and the inert filling agents are in the form of microcrystalline cellulose, Macrogol 6000, intense sweetener and a food flavourant.

EFFECT: invention provides a new tablet form of NADH.

1 ex

FIELD: medicine.

SUBSTANCE: invention refers to a pharmaceutical composition with anti-ischemic and antioxidant activity in the form of tablets or capsules, and a method for preparing it. The composition contains 4-((3-oxo-3-ethoxypropanoyl)amino)benzoic acid in an amount of 40 to 80 wt %, an amino-containing compound and pharmaceutically acceptable excipients. The amino-containing compound is specified in a group of trometamol, methyl glucamine and L-lysine; 1 mole of 4-((3-oxo-3-ethoxypropanoyl)amino)benzoic acid is accounted for 0.05 to 0.25 mole of the above amino-containing compound. The composition also contains lactose, microcrystalline cellulose, calcium stearate and other pharmaceutically acceptable excipients. According to the method for preparing the composition, taking 4-((3-oxo-3-ethoxypropanoyl)amino)benzoic acid and amino-containing compound in molar ratio 1:0.05 to 1:0.25, pre-mixing, moisturising with a aqueous or alcohol solution of a binding agent, adding pharmaceutically acceptable excipients in such an amount to provide the content of 4-((3-oxo-3-ethoxypropanoyl)amino)benzoic acid from 40 to 80 wt %, granulating the mixture, drying and producing tablets or capsules according to the known technique.

EFFECT: implementing the above application.

6 cl, 7 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: claimed is the application of 5(6)-nitro-1-(1,1-dioxothietanyl-3)-2-chlorobenzimidazole of formula (I) , earlier known as the means with broncholytic and spasmolytic activity, as the means, inhibiting peroxide oxidation of lipids.

EFFECT: realisation of the claimed purpose.

1 dwg

FIELD: chemistry.

SUBSTANCE: claimed is the application of 5(6)-nitro-2-chloro-benzimidazole of formula (I) as the preparation, inhibiting peroxide oxidation of lipids.

EFFECT: achievement of the claimed purpose, activity of the said formula compound is higher than of the comparison medication dibazole.

1 dwg

FIELD: medicine.

SUBSTANCE: method for preparing an agent possessing anti-inflammatory, diuretic and antioxidant activity, involving milling Spiraea salicifolia shoots representing a mixture of leaves, blossom and shoots, extracting them three times by gradual maceration, mixing in infusing, filtering, condensing, separating, drying in the certain environment.

EFFECT: agent shoes the pronounced anti-inflammatory, diuretic and antioxidant activity.

2 dwg, 12 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a compound of the general formula

,

wherein R1/R2 independently represent hydrogen, (CR2)o-C3-7 cycloalkyl optionally substituted by a lower alkyl or hydroxy, or represent a lower alkyl or tetrahydropyranyl, and o represents 0 or 1; and R can be identical or different, and represent hydrogen or a lower alkyl; or R1 and R2 can form together with a N atom to which they are attached, a heterocycloalkyl group specified in a group consisting of pyrrolidinyl, piperidinyl, 3-aza-bicyclo[3.1.0]hex-3-yl or 2-aza-bicyclo[3.1.0]hex-2-yl which are optionally substituted by hydroxy; R3 represents an S-lower alkyl, lower alkyl, lower alkoxy or C3-7 cycloalkyl; R3′ represents hydrogen, a lower alkyl substituted by a halogen, lower alkyl or lower alkoxy; R4 represents a lower alkyl substituted by a halogen; X represents -O- or -CH2-; X' represents -O- or -CH2-; provided one of X or X' always represent -O- and the other represents -CH2-; or a pharmaceutically acceptable acid-additive mixture, a racemic mixture, or a respective enantiomer and/or an optical isomer.

EFFECT: compounds of the general formula (I) are good inhibitors of glycine transporter 1 (GlyT-1) and hence can be used for treating schizophrenia and other neurological conditions, including pain.

13 cl, 1 tbl, 63 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to 2-pyridone compounds, represented by general formula [1], , where A represents benzene ring or pyridine ring, X represents structure, represented by general formula [3], V represents single bond or lower alkylene, W represents single bond, ether bond or lower alkylene, which can include ether bond, or their tautomers or stereoisomers.

EFFECT: obtaining pharmaceutically acceptable salts, which possess excellent activating activity with respect to GK and can be applied as medications.

27 cl, 23 tbl, 371 ex

FIELD: medicine.

SUBSTANCE: method involves a drug therapy with the antidepressant paroxetine in a daily dose of 40-50 mg a day in two stages daily for 30 days, the benzodiazepine anxiolytic diazepam 10-12 mg a day intramuscularly for 20 days, the antioxidant mexidol for the first 20 days intravenously drop-be-drop in a daily dose of 200-250 mg, and for the following 10 days in a dose of 375-400 mg orally in tablets, the immunocorrector Thymogen 0.01% in a dose of 1.2-1.3 ml intramuscularly once a day for 10 days followed by administering 1.2-1.3 ml intramuscularly every second day five times in a combination with hyperbaric oxygenation at excessive pressure 0.8-1.0 atm at compression and decompression rate 0.1 atm a minute with 40-minute isopressure from the first day of treatment for 21 days.

EFFECT: higher clinical effectiveness.

1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to 2-ethyl-6-methyl-3-hydroxypyridinium N-acetyl-aminoethanoate, which can be applied as a neurotropic preparation with anti-hypoxic, neuroprotective, anti-amnestic and vestibular-protective activity.

EFFECT: increase of the compound activity.

8 tbl, 7 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new carboxamide compounds of formula such as below, their tautomers and pharmaceutically acceptable salts, wherein the values R1, R2, R3a, R3b, R4, Q, Y, A and X are presented in clause 1 of formula .

EFFECT: compounds are cell calpain (calcium-dependent cysteine protease) inhibitors that makes them applicable for treating some diseases related to high activity of calpain.

33 cl, 2 tbl, 75 ex, 7 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to novel substance - 2-ethyl-6-methyl-3-hydroxypyridine dichloroacetate of formula I its stable crystalline form and method of obtaining thereof. Substance (I) possesses antiatherosclerotic, hypolipidemic, antihypoxic, nootropic, anxiolytic and adaptogenic action with low toxicity (LD50=30000 mg/kg, rats, intragastrically) and absence of hygroscopicity. Substance is obtained with interaction of equimolar quantities of 2-ethyl-6-methyl-3-hydroxypyridine and dichloroacetic acid in presence of solvent.

EFFECT: increase of stability.

9 cl, 8 dwg, 10 tbl, 12 ex

FIELD: medicine.

SUBSTANCE: treating cerebral oedema is the patients with chronic liver diseases is ensured by administering an ozonised solution of 6-methyl-2-ethyl-3-hydroxypyridine succinate in the ozone concentration of 1.5-7 mg/l.

EFFECT: method enables providing higher clinical effectiveness by activating aerobic paths of glucose oxidation, increasing ATP synthesis, improving tissue microcirculation, stimulating mechanisms of antioxidant protection with no side effects.

9 dwg

FIELD: veterinary medicine.

SUBSTANCE: preparation for normalisation of lipid peroxidation processes in animals comprises, wt %: 2-ethyl-6-methyl-3-hydroxypyridine succinate 20.0-30.0, ascorbic acid 5.0-7.0, selenium (Se°) 0.3-0.5, polyvinylpyrrolidone 3.0-5.0, water for injection - the rest.

EFFECT: antioxidant effect, prevention of excessive formation of lipid peroxidation products, high efficiency in normalisation of functioning of the system of antioxidant protection of animals and the process of lipid peroxidation, low toxicity and ease of administration.

6 dwg, 5 ex

FIELD: medicine.

SUBSTANCE: according to a known method of treating the liver disease accompanying type 2 diabetes mellitus involving the baseline therapy of diabetes mellitus and prescribed hepatoprotectors, the above hepatoprotector is presented by Mexicor in a daily therapeutically effective dose of not less than 16 weeks. The therapeutically effective dose of Mexicor makes 100 mg 4 times a day.

EFFECT: higher clinical effectiveness ensured by eliminating the liver disease more prominently, reducing the length of treatment, normalising the liver function test results over a short period of time, and avoiding any side effects.

2 cl, 2 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, namely to endocrinology, and can be used for treating non-alcoholic liver disease accompanying type 2 diabetes mellitus. The declared preparation Mexicor provides reducing manifestations of cytolysis and cholestasis, decreasing the steatosis index, enables improving metabolic lipid and glycaemic values and reducing insulin resistance. Mexicor is applied in a daily therapeutically effective dose of 100 mg 4 times a day for at least 16 weeks.

EFFECT: high pharmacological activity of Mexicor has been shown by achieving the pronounced and stable elimination of fatty liver disease that enables reducing the length of treatment with no side effects.

2 cl, 2 tbl

FIELD: medicine, immunology, narcology, biochemistry.

SUBSTANCE: invention elates to correction of immune disturbances in opium narcomania. Method is carried out by administration of an immunomodulating agent, 2-ethyl-6-methyl-3-oxypyridine succinate, in period for arresting opium abstinent syndrome by intravenous route, 2 times per a day in daily dose 0.1-0.4 g for 5-7 days, and at the stage of post-abstinent state - by intramuscular route in daily dose 0.1-0.2 g, 2 times per a day for 7-14 days, and at the reablement stage - by intramuscular route in daily dose 0.1-0.2 g every other day for 3-4 weeks. Method provides reducing activity of lymphocyte NO-synthase and normalization of immunocytogram picture.

EFFECT: improved method for correction.

2 ex

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