RU2360683C1 - Corrective composition for pathologic carbohydrate, lipid disbolism and antioxidant organism state involvement - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to pharmaceutical and food industry, particularly a corrective composition for pathologic lipid, carbohydrate disbolism and antioxidant organism state disorder. The corrective composition for pathologic lipid, carbohydrate disbolism and antioxidant organism state involvement representing chitosan and ecinochrome A complex, vitamins and organic acids produced from addition to dehydrated and alcohol-processed chitosan of echinochrome A, ascorbic (vitamin C), lipoic (vitamin N), citric and succinic acids in spirit.

EFFECT: composition described above is effective for correction of pathologic lipid, carbohydrate disbolism and antioxidant organism state disorder.

3 cl, 6 tbl, 5 ex

Description

The invention relates to the food industry, namely to the production of biologically active food additives (BAA) parapharmaceutical series.

An analysis of the mechanisms of development of stress-damaging effects under the influence of adverse environmental factors and conditions of professional activity showed that one of the main mechanisms in the development of disorders of homeostasis is the activation of lipid peroxidation (LPO) and a decrease in the potential of the antioxidant system (AOS). Attention to these processes is not accidental, since the activation of free-radical and peroxide processes and the so-called peroxidation syndrome underlie the pathogenesis of many diseases. In particular, it was found that peroxidation syndrome as a pathogenesis factor underlies the development of lipid and carbohydrate metabolism disorders (metabolic syndrome), which in turn lead to the development of atherosclerosis, coronary heart disease, heart attack, stroke, peptic ulcer, diabetes mellitus, etc. d. [Sidorenko B.A., Revenko V.N. Psycho-emotional stress and cardiovascular disease. // Chisinau: Stinka. 1988.194 s .; Baraboy V.A., Brekhman I.I., Golotin V.G. et al. Peroxidation and stress //. SPb .: Science. 1992. P.148]

Drug correction of disorders caused by oxidative stress, antioxidant therapy, has become in the last 10-15 years one of the leading areas in modern pharmacological and clinical studies. However, the irrational use of food-grade synthetic antioxidants ultimately led to an acute crisis. The severe long-term effects of chronic consumption of elevated doses of the main ones, ionol and butylhydroxyanisole, were reliably established. There was a need to replace the synthetic antioxidants used in the food industry and medicine with natural compounds - components of the human diet. The search for such compounds on a large scale is carried out in Japan, the USA, Finland, Switzerland and other countries. In addition, it should be noted that the use of microbial or animal antiradical enzymes as superoxide dismutase (SOD), catalase, etc. as drugs or food additives causes an inevitable response of the human immune system, and the consumption of such a universal natural antioxidant as tocopherol is limited by its E - vitamin properties. One of the most realistic approaches to the prevention of disorders of homeostasis of the body, which is based on peroxidation syndrome, is the use of dietary supplements based on natural antioxidants and enterosorbents.

Known means of rehabilitation therapy "Hitovit" containing bee bread, chitosan, rosehip powder, fucus powder and filler. The tool promotes optimal functioning and proper nutrition of human organs and tissues at the cellular level, normalizes metabolic processes [RU 2254871 C1, 2005.06.27].

Known biologically active food supplement for the prevention and adjunctive treatment of infectious diseases, containing chitosan polyfraction composition and dry extracts of phyto-harvests in the form of a complex formed by plant lectins. Supplements to food have antibacterial properties, immunocorrective action and provides a prolonging effect of phytocomponents [RU 2270585 C2, 2006.02.27].

The closest analogue of the invention, both in purpose and in composition, is a composition for normalizing lipid metabolism and reducing body weight, which is a complex comprising an equimolecular water-soluble salt of chitosan with ascorbic, succinic and lipoic acids, substances of polyhydroxynaphthoquinone nature from marine echinoderms, polyphenolic compounds from grape ridges, trivalent chromium ions and fructose [RU 2290185 C1, 2006.12.27]. To obtain this composition, chitosan is mixed with an aqueous solution of ascorbic and succinic acids, the mixture is kept under stirring until chitosan is dissolved, an aqueous-alcoholic echinoderm extract containing polyhydroxynaphthoquinones and lipoic acid is added to the resulting homogeneous solution, grape crest extract containing polyphenol compounds is added to the resulting mixture , sweetener and aqueous solution of trivalent chromium.

However, the water-alcohol extract of food commercial urchins, which is part of the additive, is not standardized. It contains mainly lipid substances and only a small amount of natural polyhydroxynaphthoquinones, the biological activity of which has not been studied. In addition, there is no reliable data on their compatibility with other dietary supplement components. It should also be noted that a large amount of fructose (up to 80%) in its composition does not put this product in the category of dietary products that contribute to metabolic correction. Water-fructose syrup must be sterilized or pasteurized, while the shelf life of such products according to the standards adopted in the Russian Federation is limited to 6 months.

The objective of the invention is to expand the range of parapharmaceuticals based on chitosan and biologically active substances of marine aquatic organisms, to obtain a biologically active additive of prolonged action, designed to correct disorders of carbohydrate, lipid metabolism and antioxidant status of the body.

As a result of solving this problem, a composition was created, which is a complex of chitosan with echinochrome A, vitamins and organic acids, obtained by adding echinochrome A, ascorbic acid (vitamin C), lipoic acid (vitamin N), citric and succinic acids to dehydrated and alcohol-treated chitosan in alcohol solution.

The content of echinochrome A in the final product is at least 0.035 wt.%.

The technical result provided by the invention is to obtain a composition that provides correction of pathological disorders of carbohydrate, lipid metabolism and antioxidant status of the body. Studies have shown that the claimed composition has anti-ulcerogenic and cardioprotective activity. Therefore, it can be used for the prevention and additional treatment of cardiovascular diseases and gastric ulcer caused by neurogenic stress.

In contrast to the prototype, the claimed composition contains a standardized polyhydroxynaphthoquinone of sea urchins - echinochrome A, the physiological effect of which is well studied (based on it, drugs of the Histochrome series are widely used in practical medicine). The compatibility of echinochrome A with other components of dietary supplements and the preservation of the biological activity of all ingredients in the composition have been experimentally shown. In addition, the claimed product is obtained in a dry state with a shelf life of at least two years.

The use of standardized echinochrome A in the composition, which belongs to the group of vitamins K, for which there is a dose recommended by WHO for humans - not more than 80-140 mcg / day, allows to standardize the final product according to GMP requirements and guarantee its safety when taken orally.

The inventive composition is obtained in the following way: to dehydrated chitosan add 96% ethyl alcohol and incubated until it is completely swollen. Citric, succinic, ascorbic, lipoic acids and echinochrome A are dissolved in 96% ethanol. A mixture of acids and echinochrome A is poured into a reaction vessel with chitosan, stirred for 30-60 minutes. The alcohol is removed by decantation, followed by purging with steam. The resulting preparation is dried.

Echinochrome A (2,3,5,6,8-pentahydroxy-7-ethyl-1,4-naphthoquinone, a vitamin of group K) - one of the pigments of sea urchins, is a multifunctional natural antioxidant [A. Lebedev, M. Ivanova M. V., Krasnovid N.I. et al. Acidic properties and interaction with superoxide anion radical echinochrome A and its structural analogues. // Questions of medical chemistry. 1999. V. 45. P.123-130] and has a high anti-radical activity, playing the role of an interceptor of reactive oxygen species [Lebedev A.V., Levitskaya E.L., Tikhonova E.V. et al. Antioxidant properties, autooxidation and mutagenic activity of echinochrome A in comparison with its structural analogues. // Biochemistry. 2001.V.66. S.885-893].

Echinochrome A (ECA) forms strong insoluble complexes with ions of variable valence metals (copper, iron), suppressing their effect as initiators of peroxidation processes [Lebedev A.V., Ivanova M.V. How do calcium ions induce free radical oxidation of hydroxyl-1,4-naphthoquinone Ca2 + stabilized the naphtosemiquinone anion-radical of echinochrome A // Arch. Biochem. Biophys. 2003. V.413. P.191-198]. It has been established that echinochrome A acts on enzyme systems involved in the development of oxidative stress (lipokinases, cyclokinases), as well as some pathological complications of aldose reductases. On its basis, drugs for cardiology and ophthalmology are created, which are introduced into medical practice under the name "Histochrome" [RU 2137472 C1, 1999.09.20; RU 2134107 C1, 1999.08.10].

For the introduction of echinochrome A into the body through the gastrointestinal tract, the authors of the invention chose chitosan. This choice was made on the basis of the following considerations. Firstly, echinochrome A interacts well with chitosan to form a complex, which avoids the sorption of echinochrome A on the mucous membranes of the digestive tract. Secondly, chitosan itself is an effective therapeutic and prophylactic agent used as an adsorbent for the treatment of many diseases. To introduce echinochrome A into a complex with chitosan, the authors use ascorbic acid.

Ascorbic acid (Vitamin C) (AK)

Ascorbic acid is one of the most widespread in nature water-soluble vitamins and is found in all organs, tissues, body fluids, especially in large quantities - in the adrenal cortex, where it is involved in metabolic processes associated with the synthesis of corticoids [Lifshits V.M. , Sidelnikova V.I. "Biochemical analyzes in the clinic." Directory. // "Medicine" M. 1993; "Nutritional supplements". Directory. // St. Petersburg. 1996]. It takes part in the implementation of redox processes (electron transport), in the formation of the main substance of connective tissue (in particular, in the hydroxylation of proline, the main amino acid component of its fibrous structures), and affects the metabolism of iron, lipids (primarily sterols), carbohydrate phosphorus metabolism, exhibits an antioxidant effect.

Ascorbic acid enters the human body only with food, because unlike other organisms in humans, primates and guinea pigs, it is not synthesized. The daily requirement of an adult in vitamin C is 50-100 mg. Insufficient intake of vitamin C in the body with food for 1-3 months leads to the initial manifestations of hypovitaminosis, after 3-6 months of the same nutrition - to a clear hypo- and vitamin deficiency, which is manifested by metabolic disorders of many substances, dysfunction of the central nervous and endocrine systems ( weakness, irritability, pain in the extremities), a decrease in resistance to infections, a violation of regenerative processes and the development of scurvy (scout). Vitamin C deficiency is usually accompanied by a deficiency of vitamin R.

In medical practice, ascorbic acid is used to treat hypovitaminosis, stimulate hematopoiesis together with folic acid, vitamin B and iron, to strengthen capillaries with increased bleeding in various diseases, stimulate regenerative processes, lesions of the connective tissue, diseases of the respiratory tract, etc.

Vitamin N (lipoic acid) (LK) is found in many plant and animal organisms in the form of a protein bound ε-lipolysin and is a coenzyme of multienzyme complexes pyruvate dehydrogenase, α-ketoglutarate hydrogenase, which carry out oxidative decarboxylation of α-keto acids and the construction of coenzyme acyl It is involved in the regulation of protein, lipid and carbohydrate metabolism, exhibits a lipotropic effect, affects cholesterol metabolism, improves liver function, has antidote properties, and malizes energy processes ["Nutritional Supplements". Directory. // St. Petersburg. 1996; Pilate T.L., Ivanov A.A. Supplements for food. // Moscow. 2002]. Lipoic acid is widely used in medicine to normalize lipid metabolism, treat certain liver diseases (cirrhosis, Botkin’s disease), diabetes mellitus, atherosclerosis, some poisoning, as well as in pediatrics and in case of visual impairment. Lipoic acid protects the liver from the toxic effects of alcohol. It can also be used as a chelating agent, especially for removing excess copper from the body. The need for lipoic acid is approximately 0.5-2.0 g per day. Specific manifestations of deficiency are not described.

Citric acid (E33O) (LMK) is a well-known acidity regulator, antioxidant and synergist of antioxidants, a complexing agent, dispersant and disintegrant ["Food additives". Directory. // St. Petersburg. 1996]. Citric acid is the mildest compared to other food acids to taste. It has a pleasant sour taste, due to which it is widely used in the food industry. To the greatest extent, citric acid is used in the confectionery industry and in the production of soft drinks, as well as in the production of certain types of canned fish. The advantage of citric acid is the absence of irritating effects on the mucous membranes of the digestive tract.

Succinic acid (UC) improves the processes of energy metabolism in the cells of the brain, myocardium, liver and kidneys. It has a pronounced antihypoxic and detoxicating effect, accelerates cell energy recovery processes ["Nutritional Supplements". Directory. // St. Petersburg. 1996]. As a food acid (acidity regulator), it is allowed in all countries - E363.

Chitosan, obtained from the natural chitin polysaccharide by alkaline deacetylation, is a polysaccharide consisting of D-glucosamine residues linked by β-1,4-glycosidic bonds [Chitosan per os (translated from English) // Edited by A. Riccardo. Muzzarelli. Nizhny Novgorod. Publishing house "Vector - TiS", 2002 - 372 s]. Calcination of chitosan at 170 ° C leads to the removal of water molecules and the formation of free amino groups.

Chitosan is an excellent polycationic adsorbent. Due to this property, it has found wide application in medicine both as an enterosorbent and as a carrier for drug delivery. The amino groups of chitosan easily form salts with mineral and organic acids and other substances that have an acidic function, for example, with vitamin C.

In the production of the claimed composition, the reaction of chitosan with echinochrome A and the above ingredients (organic acids and vitamins) is carried out in a heterogeneous phase according to the following scheme:

As a result of the reaction, a complex is obtained having the following structure:

где R^-=ЭХА^-, АК^-, ЛК^-, ЛМК^- и ЯК^-

where R ^- = ECA ^- , AK ^- , LC ^- , LMK ^- and UC ^-

The invention is illustrated by the following examples.

EXAMPLE 1

600 g of food or medical chitosan are dehydrated by calcination in an oven at a temperature of 170 ° C for 15-20 hours and placed in a round-bottom flask of a rotary evaporator. To chitosan add 1-2 l of freshly distilled ethyl alcohol 96 ° until the chitosan completely swells and an alcohol “mirror” is obtained over chitosan. The selected organic acids (150 g citric acid, 100 g succinic acid), vitamins (45 g ascorbic acid, 75 g lipoic acid and 0.35 g echinochrome A) are dissolved in 96 ° freshly distilled ethanol and poured into a reaction flask with chitosan and stirred within 30 minutes The flask with the mixture is placed on a rotary evaporator, completely ethanol is removed from the reaction mixture in vacuum at a temperature not exceeding 60 ° C. A dry powder is obtained, which is encapsulated in gelatin capsules of 1.5 ml or tablets of 0.25 and 0.5 g.

The composition contains, wt.%: Chitosan - 60.0; ascorbic acid - 4.5; echinochrome A - 0.035; lipoic acid - 7.5; succinic acid - 10.0; citric acid - 15.0; residual moisture - 4.0.

EXAMPLE 2

6.2 kg of food or medical chitosan is dehydrated by calcination in an oven at a temperature of 170 ° C for 15-20 hours and placed in a reactor with a stirrer. To chitosan add 15-20 l of freshly distilled ethyl alcohol of 96 ° until the chitosan completely swells and an alcohol “mirror” is obtained over chitosan. Organic acids (1.5 kg of citric acid, 1 kg of succinic acid) and vitamins (430 g of ascorbic acid, 770 g of lipoic acid and 4.0 g of echinochrome A) are dissolved in a separate container in 96 ° freshly distilled ethyl alcohol, and poured into the reaction vessel with chitosan and stirred for 60 minutes. Alcohol from the reactor is removed by decantation, followed by purging with steam. The resulting preparation is dried in a shelf-type oven for 12-24 hours at a temperature not exceeding 110 ° C. Supplements are Packed in glass jars of 250-500 g or tableted at 0.25 and 0.5 g.

The composition contains, wt.%: Chitosan - 62.0; ascorbic acid - 4.3; echinochrome A - 0.04; lipoic acid - 7.7; succinic acid - 10.0; citric acid - 15.0; residual moisture is 1.0.

EXAMPLE 3

The study of corrective activity of dietary supplements on a model of nutritional hyperlipoproteinemia (HFP)

The alimentary GLP model was reproduced by unbalancing the diet according to the content of proteins, carbohydrates, fats, and cholesterol - as a result, animals on the experimental diet developed hypercholesterolemia (GHS), hypertriglyceridemia (GTG). The content of atherogenic lipoproteins in blood - fractions of low and very low density lipoproteins (LDL, VLDL) increased more than twice. Changes in the lipid background could be characterized as a manifestation of one of the most common types of HFD - type IV according to the WHO classification.

It is known that lipid peroxidation (lipid peroxidation) is enhanced during HFL. Excessive intake of high-calorie foods and unbalanced nutrition contribute to the intensification of lipid peroxidation processes in the blood, as can be judged by an increase of 60% (p <0.01) in the content of malondialdehyde (MDA) in red blood cells. The response of the antioxidant defense system (AOD) was expressed in an increase in the activity of the enzyme glutathione reductase (GR), more than three times, resulting in an increase in the content of the reduced form of glutathione (GL), necessary to neutralize peroxides. The mobilization of all components of the AOD system in response to increased LPO processes during HFL is also proved by a decrease in the integral indicator of the state of AOD - the level of antioxidant activity (AOA) of the blood.

The enhancement of lipid peroxidation in the liver was confirmed by an increase in the content of its products at all stages of the peroxide cascade (tables 1, 2). The marked inhibition of glutathione peroxidase (GP) and GR activity, a decrease in the level of reduced GL and antioxidant vitamins testified to a decrease in AOZ reserves in response to increased LP. Activation of lipid peroxidation processes in the liver is accompanied by the suppression of cholesterol (cholesterol) catabolism in hepatocytes, which, in turn, helps to maintain elevated plasma cholesterol levels. The inclusion in the diet of funds that enhance the reserves of AOZ of the body can inhibit the development of HFL and correct the occurring lipid metabolism disturbances [Lankin V.Z., Vikhert A.M., Tikhadze A.K. et al. // Vopr honey chemistry. 1989. No3. S.18-24; Riccvuti G., Mazzone A., Pasotti D., Deservi S., Specchia G.I. // Atherosclerosis. 1991. V. 91. P.1 .; Avtsin A.P., Zhavoronkov A.A., Rish M.A., Strochkova L.S. // "Human microelementoses." "Medicine" M. 1991; Semenov N.V. // "Biochemical components and constants of liquid media of human tissues." "The medicine". M. 1971].

Intragastric administration of the claimed composition to animals revealed a decrease in the rate of formation of lipoperoxides in the liver, an increase in the activity of glutathione-dependent enzymes, an increase in the content of vitamin A and α-tocopherol with statistical significance of changes compared with untreated animals with HF (tables 1, 2).

An improvement in the state of blood lipid background was confirmed by a decrease in total cholesterol (OXC) by 55% (p <0.01) and triglycerides (TG) by 61% (p <0.01). The content of VLDL and LDL decreased significantly (by 63% and 35%, respectively, p <0.01).

A positive point was the increase in cholesterol (cholesterol) in high density lipoproteins (HDL). A significant increase in the activity of glutathione peroxidase (GP) against the background of a decrease in reduced glutathione (GL) indicates an active reaction to neutralize peroxides in red blood cells, which protects them from hemolytic decomposition.

Table 1 Indicators of lipid metabolism and the system of lipid peroxidation - AOD of the blood of animals with HFP treated with the claimed composition (M ± m) Indicators 1st group control 2nd group of GLP 3rd group of GLP + dietary supplement OHS, mmol / l 1.27 ± 0.02 2.92 ± 0.06 * 1.30 ± 0.12 ** HS VLDL, mmol / l 0.24 ± 0.01 0.59 ± 0.02 ** 0.22 ± 0.01 ** HSLPNP, mmol / l 0.91 ± 0.02 2.17 ± 0.08 ** 1.40 ± 0.07 ** HDL cholesterol, mmol / l 0.11 ± 0.04 0.16 ± 0.03 0.3b ± 0.05 ** TG, mmol / l 0.56 ± 0.01 1.29 ± 0.09 ** 0.50 ± 0.02 ** GL, mmol / g protein 4.58 ± 0.46 9.40 ± 2.15 * 4.61 ± 0.44 * GR, mmol NADP / min / g Hb 26.64 ± 4.66 92.97 ± 21.89 * 40.13 ± 7.84 GP, μmol / min / mg Hb 49.95 ± b, 02 39.96 ± 4.75 99.32 ± 4.88 ** MDA, mmol / g protein 7.85 ± 0.89 12.55 ± 0.57 ** 8.96 ± 0.99 AOA,% 62.50 ± 9.22 59.23 ± 5.21 * 83.32 ± 4.81 * Note. Assessment of significance of differences between groups: 1/2, 3/2 * -p <0.05, ** - p <0.01 table 2 Indicators of the LPO-AOP system in the liver of animals with HFP treated with the claimed composition (M ± t) Indicators 1st group control 2nd group of GLP 3rd group of GLP + dietary supplement GL, nmol / mg total protein 62.32 ± 1.02 42.86 ± 1.03 58.20 ± 0.82 GR, nmol NADP / mg protein / min 1.79 ± 0.14 0.77 ± 0.04 1.12 ± 0.01 GP, nmol GL / hour / mg protein 76.17 ± 1.92 56.00 ± 0.98 65.26 + 1.02 DC, nmol / mg lipids 4.95 ± 0.08 9.66 ± 0.25 6.83 ± 0.24 MDA, nmol / mg protein 1.77 ± 0.09 4.76 ± 0.22 3.20 ± 0.15 Vitamin A, ng / mg lipid 71.05 ± 2.78 54.40 ± 2.73 63.33 ± 2.2 * α-tocopherol, ng / mg lipid 173.15 ± 4.06 67.50 ± 2.30 97.03 ± 2.25 Note. Assessment of significance of differences between groups: 1/2, 3/2 p <0.01; * p <0.05

A 40% increase in the antioxidant activity (AOA) of the blood is also indicative of an increase in the reserves of the AOD system (Table 1).

Thus, the lipid-lowering and antioxidant effect of the claimed composition allows us to recommend it as a prophylactic for lipid metabolism disorders, characterized by the development of hyperlipoproteinemia (HLP), hypercholesterolemia (HCS) and hypertriglyceridemia (GTG).

EXAMPLE 4

Evaluation of the antihyperglycemic activity of dietary supplements on the model of alloxan diabetes

Pharmacological studies in an experimental model of alloxan diabetes were carried out on white female mice of the CD-1 line weighing 20-22 g. Animals were previously left without food for 15 hours. Alloxan diabetes was caused by a single intraperitoneal injection of alloxan at a dose of 120 mg / kg in isotonic solution. The inventive composition was administered orally at 10-fold dilution for 4 days before and 4 days after the induction of diabetes. The therapeutic and prophylactic effect was determined 10 days after the start of the experiment using a glucose tolerance test. To this end, animals on an empty stomach were orally administered 4 g / kg of an aqueous solution of glucose, and after 60 min, blood was taken. OneTouch Ultra glucometer (Germany) was used to analyze the concentration of glucose in the blood, and Novogluk-K, M diagnostic kit (Vector-Best, Russia) was used to determine the enzymatic activity of aspartate aminotransferase (AsAT), alanine aminotransferase (AlAT) in blood plasma. In addition to the experimental groups of treated animals, the experiment included a group of intact animals and a group of untreated negative control with alloxan diabetes induced by the general scheme. The state of the antioxidant defense system in diabetic animals was determined by the level of LPO products [Karpishchenko A.I. Medical laboratory technology. - St. Petersburg: Inter-Medica, 1999].

Alloxan diabetes in laboratory animals is a model of insulin-dependent diabetes mellitus, in the pathogenesis of which the progressive death of pancreatic β-cells plays a key role. The cause of this death is “oxidative stress” induced at the cellular and molecular level and, as a result, insufficient insulin synthesis. One of the striking manifestations of disorders of tissue metabolism reactions in diabetes is the intensification of the processes of free radical lipid oxidation, which is maintained in the physiologically metabolizing organism within strictly limited boundaries, which is ensured by the actively regulated balance in the norm of two countersystems - mechanisms of pro- and antioxidant effects. A significant role in this is assigned to the components of the endogenous system of antioxidant defense of the cell [Burlakova EB, Dzhalyabova MI In: Bioantioxidants in the regulation of metabolism in normal and pathological conditions. M. 1982. S. 113-141].

During the experimental work, the antidiabetic drug Metformin (Berlin-Chemie, Germany) was used as a reference drug. The results are presented in table 3

Table 3 The effect of the claimed composition and "Metformin" on clinical and diagnostic indicators of the pancreas and blood plasma of experimental animals with alloxan diabetes No. p / p Group of animals Plasma glucose, mmol / l TBA reactive products, mcmol / g tissue Diene conjugates, µmol / g tissue AlAT, µmol / l hour AsAT, µmol / l hour Bilirubin, μmol / L one Intact 3.8 ± 0.3 1.1 ± 0.1 5.5 ± 0.5 2.9 ± 0.2 3.5 ± 0.3 56.7 ± 4.2 2 Alloxan, (-) control 10.5 ± 0.8 3.8 ± 0.4 15.4 ± 1.4 4.9 ± 0.4 5.3 ± 0.5 86.6 ± 6.8 3 The inventive composition, 10-fold dilution 5.2 ± 0.5 1.6 ± 0.1 7.4 ± 0.7 3.3 ± 0.3 4.1 ± 0.4 64.8 ± 6.1 four Metformin 10 mg / kg 4,5 ± 0,4 2.1 ± 0.2 12.6 ± 1.1 3.4 ± 0.3 4.1 ± 0.4 62.4 ± 6.4 Note: Results are presented as mean ± standard deviation (p <0.05).

As can be seen from the results presented in table 3, alloxan diabetes was accompanied by hyperglycemia - 10.5 ± 0.8 mmol / L (in the intact control - 3.8 ± 0.3 mmol / L; p <0.05), which can be considered as a manifestation of insulin deficiency and activation of gluconeogenesis.

Carrying out the treatment and prophylactic course against the background of alloxan diabetes with the claimed composition (10-fold dilution) and Metformin at a dose of 10 mg / kg led in both cases to a noticeable (about 2-fold) decrease in glucose compared with the negative control. Thus, the claimed composition on the model of alloxan diabetes exhibits pronounced corrective activity.

“Oxidative stress” caused by alloxan leads to a marked increase in plasma aminotransferases and bilirubin, as well as a marked increase in the level of TBA-reactive products and diene conjugates in the pancreas. Conducting a treatment course with a curled composition against alloxan diabetes leads to a significant restoration of pancreatic function and redox processes in its tissues and to a pronounced normalization of biochemical blood parameters (table 3).

EXAMPLE 5

The study of anti-ulcerogenic and cardioprotective activity of dietary supplements

The pronounced antioxidant properties of the claimed composition suggested that it will have a prophylactic effect in the development of diseases caused by neurogenic stress. Therefore, we have studied its antiulcerogenic and cardioprotective properties in an experimental model of neurogenic stress.

The antiulcer effect of the claimed composition was studied on white outbred mice weighing 20 ± 2 g. Acute stress was modeled by suspending the animals by the neck fold for 22 hours. The studied drug was administered per os using a probe for 6 days and on the seventh day 1 hour before suspension in 5-, 10- and 20-fold dilutions of the original preparation in a volume of 200 μl. Control animals were injected with an equivalent volume of solvent. The antiulcer effect of the claimed composition was evaluated according to [Zueva EP, Krylova SG, Turetskova V.F. et al. Experimental study of antiulcer properties of aspen bark extract. // Experimental and clinical pharmacology. 1997.V.60. S.38-41].

The cardioprotective effect of the claimed composition was studied on a model of neurogenic stress, as described above. For biochemical analyzes in animals, blood was taken by decapitation in the suborbital region. Destructive damage to the heart muscle was determined by measuring the activity of the enzymes aspartate aminotransferase (AsAT) and alanine aminotransferase (AlAT) in blood serum using the diagnostic kits of the company Vector-Best (Novosibirsk).

To assess the overall protective effect of dietary supplements, the ratio of the mass of lymphoid organs of the spleen and thymus to the body weight of animals was determined and expressed as a percentage of the norm (intact animals).

The antioxidant status of the animal organism was assessed by the activity of catalase in blood red blood cells and the level of the end-product LPO malondialdehyde (MDA). The optical density was determined on a CECIL 1011 spectrophotometer (BRUKER, Germany).

Table 4 presents data on the antiulcer activity of the claimed composition against the background of injuries of the gastric mucosa of experimental animals induced by acute stress. The gastroprotective effect of the drug was manifested in a decrease in the number of animals with ulcers and a decrease in the average number of ulcers per animal. If in the control group 100% of the animals had lesions of the gastric mucosa, then the introduction of the claimed composition reduced this indicator to 60%, 0% and 25% at 5, 10 and 20-fold dilution of the drug, with the average number of ulcers per animal compared to control decreased by 54%, 100% and 10%, respectively.

Table 4 The influence of the claimed composition on the development of ulcerative destruction of neurogenic origin Groups of animals Dilution of the drug The number of animals in the group The number of animals with destruction of the mucosa,% The average number of ulcers per animal I - 5 0 0 II - 5 one hundred 8.8 III 5 5 60 6.8 IV 10 5 0 0 V twenty four 25 8.0 Designations: I - intact animals, II - control group of animals, III-V - experimental groups of animals that received the claimed composition with 5-, 10- and 20-fold dilutions, respectively.

The inventive composition has a pronounced anti-ulcerogenic effect. In all animals of the experimental group (negative control), a decrease in hemorrhagic inflammation and a decrease in the intensity of ulcerative lesions were observed. The most pronounced effect was shown with a 10-fold dilution of the drug, in which the animals showed a complete absence of ulceration.

It is known that the content of aminotransferases in blood plasma increases sharply in some pathological conditions, in particular, accompanied by destructive processes in parenchymal organs. Despite the lack of organ specificity, the determination of aspartate aminotransferase (AsAT) and alanine aminotransferase (AlAT) for heart diseases has great diagnostic value [Menshikov VV, Delektorskaya LN, Zolotnitskaya RP // Laboratory methods of research in the clinic: Reference. M .: Medicine. 1987. S.368-370]. The results are presented in table 5.

Table 5 Determination of the enzymatic activity of AsAT and AlAT in intact animals and animals exposed to neurogenic stress Observation group Dilution of the drug AlAT, μmol / l · h AsAT, μmol / l · h I - 4.28 7.64 II - 6.58 9.38 III 5 3.9 6.56 IV 10 4.2 6.54 V twenty 4.76 4.04 Designations. I - intact animals, II - control group of animals (negative control), III-V - experimental groups of animals that received the claimed composition with 5-, 10- and 20-fold dilutions, respectively. AlAT - alanine aminotransferase, AcAT - aspartate aminotransferase.

As follows from table 5, the average level of activity of AsAT in the blood serum of control animals as a result of stress increases by 1.2 times compared with the activity of AlAT. The predominant increase in ACAT activity compared to ALAT reflects a destructive lesion of the muscle tissue of the heart. A clear tendency to normalize the activity of aminotransferases in the blood serum of animals treated with the drug as a preventive measure (before exposure to stress) suggests its cardioprotective effect. The greatest protective effect of the claimed composition is observed when it is used in 10-fold dilution.

In the process of experimental work, the effect of the claimed drug on the antioxidant status of animals exposed to neurogenic stress was also studied. In addition, we studied the dynamics of changes in the mass of the lymphoid organs of the immune system - the thymus and spleen, to assess the overall protective effect of dietary supplements. The results are presented in table 6.

Table 6 An experimental study of the antioxidant protective effect of the claimed composition Group of animals Dilution of the drug MDA,% Catalase,% Thymic index Splenic index I - 32 47 2,32 7.18 II - 44 34 0.91 2,55 III fifty 35 47 1,02 4.06 IV one hundred 24 46 1.42 4.21 V 200 24 51 1.31 3.87 Designations: I - intact animals, II - group, negative control, III-V - experimental groups of animals that received the claimed composition with 5-, 10- and 20-fold dilutions, respectively. MDA is malondialdehyde.

As can be seen from table 6, the prophylactic use of this composition in an experimental model of neurogenic stress partially inhibits the involution of the thymus and spleen, which may indicate its protective effect on the immune system.

In the serum of intact mice there is a certain level of end products of free radical oxidation, in particular MDA. In untreated animals subjected to neurogenic stress, the MDA level increases by 1.4 times, which indicates the induction of the formation of free radicals as a result of stress.

The prophylactic use of the claimed composition in an experimental model of neurogenic stress gives not only a pronounced therapeutic effect, but is also accompanied by a decrease in the products of peroxidation in the blood plasma of experimental animals compared with the control. A clear tendency to normalize the level of catalase activity is observed in animals of all groups treated with the composition. The normalization of free radical processes in treated animals may be due to direct antioxidant action (the ability of individual components of the claimed composition to capture and neutralize free radicals formed in the process of damage to the stomach, heart and other organs) and / or by acting at the cellular level on the activity of antioxidant enzymes of these organs and is mediated by a general increase in body resistance.

Analysis of the results allows to identify a direct correlation between the positive cardioprotective effect of the claimed composition and its antioxidant properties.

Claims (3)

1. Composition for the correction of pathological disorders of lipid, carbohydrate metabolism and the antioxidant status of the body, which is a complex of chitosan with echinochrome A, vitamins and organic acids, obtained by adding echinochrome A, ascorbic acid (vitamin C), lipoic acid (vitamin C) to dehydrated and alcohol-treated chitosan N), citric and succinic acids in an alcohol solution. 2. The composition according to claim 1, where the content of echinochrome A in the final product is not less than 0.035 wt.%. 3. The composition according to claim 1 or 2, which can be used for the prevention and additional therapy of cardiovascular diseases and gastric ulcer caused by neurogenic stress.