RU2519744C2 - Pharmaceutical composition, medication for prevention and treatment of metabolic syndrome and diabetic nephropathy and method of obtaining thereof - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to pharmaceutical industry and represents a pharmaceutical composition for prevention and treatment of the metabolic syndrome and diabetic nephropathy, which contains the following active substances: taurine, a dry extract of motherwort herb, a dry extract of hawthorn fruit and accessory substances, with components present in the composition in a specified ratio in wt %.

EFFECT: invention provides extension of the arsenal of means for prevention and treatment of the metabolic syndrome and diabetic nephropathy.

10 cl, 13 ex, 19 tbl

Description

The invention relates to medicine and the pharmaceutical industry and, more specifically, to a pharmaceutical composition, agent and method for its preparation for the prevention and treatment of metabolic syndrome and diabetic nephropathy and can be used to produce on their basis a drug in the form of tablets, film-coated tablets , or capsules.

A well-known pharmaceutical composition containing the active substances taurine, motherwort herb extract, hawthorn fruit extract and excipients is described in the Ukrainian patent as a Kratal agent for treating diseases of the cardiovascular system (Ukrainian patent No. 39141, IPC АКК 31/195, 35/78, 2001). The known composition is intended to obtain granules containing taurine and thick extracts of the fruits of hawthorn and motherwort, obtained by wet granulation with potato starch, followed by drying.

Known tool Kratal for the treatment of diseases of the cardiovascular system, containing active substances - taurine, motherwort herb extract, hawthorn fruit extract and excipients (Ukrainian patent No. 39141, IPC АКК 31/195, 35/78, 2001). The composition of Kratal granules is selected in the following ratio of components, wt.%:

Taurine 85.0-86.8 Thick motherwort herb extract 8.3-8.7 Thick Hawthorn Fruit Extract 4.1-4.4 Excipients rest

Due to its pharmacotherapeutic properties, the combined Kratal granule agent has a mild cardiotonic effect, antianginal, coronarolytic and antiarrhythmic activity. The mechanism of the pharmacotherapeutic action of the drug is associated with an inhibitory effect on the renin-angiotensin and kallikrein-kinin system with a simultaneous positive effect on the production of cAMP and inhibition of lipid peroxidation.

However, the granules have unpleasant organoleptic properties (extremely pronounced bitterness) when administered orally. The dosage of the drug in granules is also ambiguous and difficult not only for adults, but especially when prescribed to children.

The technology for producing this composition is very problematic, since wet granulation with starch paste of all active components in order to obtain granules, subsequent drying operations, then dry granulation by grinding through sieves and, finally, classification of granules is quite energy-consuming and long-term operation. In addition, the use of starch paste as an agent for granulating water-soluble active ingredients - taurine and thick extracts of plant substances of hawthorn fruit (Crataegus janguinca) and motherwort grass (Leonurus cardiaca L.) is a rather difficult to reproduce technological operation.

An important pharmaceutical problem for Kratal is the selection of reliable blister packs due to the significant hygroscopicity of this drug. The widely used blister based on a polyvinyl chloride film with an aluminum substrate is not always a reliable tool as a hydro-barrier for the preservation of drugs such as Kratal for a long storage time (more than 1 year, pharmacopoeial requirements) even in a narrow range of humidity indicators. In such a package, Kratal, even at room temperature and with an air humidity of 60-70%, does not withstand 2-2.5 months of storage. As a result, the tablets acquire a dark brown color and become elastic, and when lightly exposed to mechanical stress when removed from the blister, they undergo deformations, like rubber.

In accordance with UA patent 39141, Kratal granules are intended for the treatment of the cardiovascular system, which is a very broad definition of the possibilities of medical use of this drug with an ambiguous understanding of its therapeutic efficacy. In the Instructions for medical use for the drug Kratal produced by the NPC “Borshchagovsky Chemical Farm” (registration UA 3866/01/01 until 10/29/2015), the treatment indications are “neurocirculatory dystonia, as part of combination therapy for chronic coronary artery disease, post-radiation syndrome”.

The inventions are based on the task of creating such a pharmaceutical composition, means and method for its production for the prevention and treatment of metabolic syndrome and diabetic nephropathy, in which the choice of a new quantitative and qualitative composition of the components in the composition and the agent based on it, and new conditions for performing operations in the method was would be a correction in relation to the components of the metabolic syndrome - mitochondrial bioenergetic processes of the cardiovascular system, prevention and treatment iabeticheskoy nephropathy.

The problem is solved in that in the pharmaceutical composition for the prevention and treatment of metabolic syndrome and diabetic nephropathy containing the active substances taurine, motherwort herb extract, hawthorn fruit extract and excipients, according to the invention, the components are taken in the following ratio wt.%:

Hawthorn extract liquid * or thick * or dry 2-5 Motherwort extract liquid * or thick * or dry 6-10 Taurine 65-80 Excipients rest

* - (in terms of dry matter)

In addition, dry extracts of hawthorn and motherwort are introduced into the composition in the form of a complex with excipients.

It is advisable that the tool for the prevention and treatment of metabolic syndrome and diabetic nephropathy, containing the active substances taurine, motherwort herb extract, hawthorn fruit extract and excipients, according to the invention contained a pharmaceutical composition in the following ratio, wt.%:

Hawthorn extract liquid * or thick * or dry 2-5 Motherwort extract liquid * or thick * or dry 6-10 Taurine 65-80 Excipients rest

* - (in terms of dry matter)

In addition, substances that act as binders, disintegrating agents, and materials that facilitate the sliding and lubrication of granules during pressing, and film coating materials are selected as auxiliary substances.

In addition, as substances that act as binders, the tool contains starch potato or microcrystalline cellulose to obtain granules for tabletting or encapsulation.

In addition, as agents causing disintegration, the product contains croscarmellose sodium to obtain granules with liquid, or thick, or dry plant extracts.

In addition, as materials that contribute to the sliding of the granulate during pressing and film coating materials, the product contains colloidal silicon dioxide.

In addition, as materials that promote lubrication, the tool contains magnesium stearate, introduced into the composition of dry complexes with plant extracts and upon receipt of granules for tabletting and encapsulation.

In addition, the tool contains components in the form of dosage forms of tablets or tablets, film-coated, or capsules at a daily dose of 100 mg per taurine per kg of body weight.

It is advisable that the method of obtaining funds for the prevention and treatment of metabolic syndrome and diabetic nephropathy was characterized by the fact that previously liquid, thick or dry extracts of hawthorn fruit and motherwort grass were obtained by extraction with an aqueous or aqueous-ethanol solvent by maceration or percolation, filtration extraction, or repercolation followed by vacuum concentration for a thick extract and / or subsequent drying for a dry extract, after which they were mixed with taurine and excipients that perform the functions of binders, disintegrating agents, and materials that promote the sliding and lubrication of granules during pressing.

The pharmaceutical development of dosage forms was based on the use of a pharmaceutical composition with a constant composition of pharmacologically active components at various ratios of excipients using various technological methods.

In preclinical studies, a pronounced antioxidant, antihyperglycemic, hypolipidemic vasodilator, antisclerotic, antiplatelet and antitoxic effect of the new drug was found. It turned out that the revealed pharmacological properties of the drug, aimed at the correction of metabolic disorders induced by insulin resistance and insulin deficiency, are realized by reducing oxidative stress and mitochondrial dysfunction, which plays a key role in the pathogenesis of the above diseases. It should be noted that the normalization of bioenergy processes due to the restoration of mitochondrial function and oxidative status occurs due to the synergism of the pharmacological action of all components of the drug.

In the manufacture of the pharmaceutical composition in the dosage form of tablets or capsules, one or more pharmaceutically acceptable excipients have been used, which act as binders, disintegrating agents, materials that promote the sliding and lubrication of granules during compression.

As binders, potato starch and microcrystalline cellulose were preferred. According to the present invention, the amount of binder material can range from 1 to 50%, preferably from 5 to 25%, based on the total weight of the tablet. The composition of the tablet or capsule according to the binder components was selected so as to provide an acceptable oral dosage form and convenient for the patient with the immediate release of the active ingredients.

Preference was given to croscarmellose sodium and crospovidone as disintegrating agents. The amount of disintegrant can vary in the range from 0.5 to 20%, it has become preferable from 1 to 10%.

Colloidal silica was found to be the preferred agent for sliding the components of the granulate. The amount of component for sliding can vary in the range from 0.1 to 10%, preferably 0.25 to 5% is selected.

Magnesium stearate was used as a lubricant component. The amount of the lubricant component can vary in the range from 0.1 to 5%, from 0.5 to 2% has become preferable.

As materials for coating the tablets with a shell, products such as Opadry® from Colorcon, Advantia® from ISP, Sepifilm® from Seppic, which turned out to be pharmaceutically acceptable components, were used. The amount of film coating can vary in the range from 1 to 10%, from 1.5 to 5% turned out to be acceptable.

According to pharmaceutical development, due to the varying degrees of hygroscopicity of dry extracts, their complexes with excipients and some types of tablets, the use of appropriate packaging was required. When testing dosage forms in various moisture and temperature conditions with a long storage time (pharmacopoeial requirements), the most acceptable were plastic containers for tablets and capsules, blister packs of the polymer / metal type, namely PVC / PVDC / Alu composite (polyvinyl chloride-polyvinyl dichloride-aluminum) or Alu / Alu.

The established combination of pharmacotherapeutic properties of the drug allows us to predict the prospects for the use in the prevention and treatment of metabolic syndrome, type 2 diabetes mellitus, its micro- and macrovascular complications, and diabetic nephropathy.

Example 1

Obtaining a dry complex containing extracts of hawthorn or motherwort, microcrystalline cellulose and silicon dioxide colloidal, the following component composition:

Component of the dry complex Content% Dense hawthorn or motherwort extract (dry matter content 60%), calculated on the dry extract 70.0 Microcrystalline cellulose 25.0 Silicon Colloidal Dioxide 5,0 Total: one hundred

Dense extracts containing 60% solids are placed in a stirred reactor. During mixing, microcrystalline cellulose and silicon dioxide are added, colloidal dioxide, then the homogeneous mixture is dried in a vacuum belt dryer and crushed. The granulate obtained after the addition of excipients is used for tabletting or encapsulation.

Example 2

The preparation of a dry complex differs from Example 1 in that microcrystalline cellulose is introduced into the thick extracts with stirring before drying to obtain the following component composition:

Component of the dry complex Content% Dense hawthorn or motherwort extract (dry matter content 60%), calculated on the dry extract 70.0 Microcrystalline cellulose 30,0 Total: one hundred

Example 3

The preparation of a dry complex differs from Examples 1 and 2 in that potato starch and colloidal silicon dioxide are introduced into the thick extract with stirring before drying to obtain the following component composition:

Component of the dry complex Content% Dense hawthorn or motherwort extract (dry matter content 60%), calculated on the dry extract 72.0 Potato starch 25.0 Silicon Colloidal Dioxide 3.0 Total: one hundred

Example 4

It differs from Examples 1, 2 and 3 in that potato starch is introduced into the thick extract with stirring before drying to obtain the following component composition:

Component of the dry complex Content% Dense hawthorn or motherwort extract (dry matter content 60%), calculated on the dry extract 75.0 Potato starch 25.0 Total: one hundred

Example 5

The preparation of the dry complex differs from Examples 1, 2, 3 and 4 in that the thick extracts of hawthorn and motherwort are mixed in a mass ratio of 1: 2, then microcrystalline cellulose and silicon dioxide colloidal are added to the resulting mixture with stirring before drying to obtain the following component composition:

Product Content% Dense hawthorn extract (dry matter content 60%), calculated on the dry extract 23.3 Thick motherwort extract (dry matter content 60%), calculated on the dry extract 46.7 Microcrystalline cellulose 25.0 Silicon Colloidal Dioxide 5,0 Total: one hundred

Several methods have been developed according to the invention for the preparation of the particular desired dosage form — tablets, coated tablets, or capsules.

Example 6

The pharmaceutical composition in the dosage form of tablets is obtained by mixing all the components in dry form, pressing the mixture with the composition of all components in weight mass and mass%.

The composition of the dosage form in tablets:

Component No. Composition per 1 tablet (mg) % Dry hawthorn extract (6.1) 43 3.91 Dry motherwort extract (6.1) 87 7.91 Taurine (6.2) 867 78.82 Microcrystalline cellulose (6.2) 70 6.36 Croscarmellose sodium (6.3) eleven 1.00 Silicon Colloidal Dioxide (6.3) eleven 1.00 Magnesium stearate (6.3) eleven 1.00 total weight 1100 100.00

Example 7

It differs from Example 6 in that the tablets are obtained using thick extracts of motherwort and hawthorn with a moisture content of up to 50% by weight. By wet granulating the mixture of components (7.2) with extracts (7.1), drying the obtained granulate, calibrating the granulate, mixing with the components (7.3), compressing the new granulate, the resulting tablets are packaged in a blister pack.

The composition of the dosage form in tablets:

Component No. Composition per 1 tablet (mg) % Dense hawthorn extract (dry matter content 50%), calculated on the dry extract (7.1) 43 3.91 Motherwort extract is dense (dry matter content is 50%), calculated on the dry extract (7.1) 87 7.91 Taurine (7.2) 867 78.82 Microcrystalline cellulose (7.2) 70 6.36 Croscarmellose sodium (7.3) eleven 1.00 Silicon Colloidal Dioxide (7.3) eleven 1.00 Magnesium stearate (7.3) eleven 1.00 total weight 1100 100.00

Example 8

In this method, the preparation of tablets for children differs from Examples 6 and 7 in that the tablets are half the dosage and are coated with an aqueous dispersion of the film mixture (8.4).

Composition of tablets for children:

Component Composition per 1 tablet (mg) % Thick hawthorn extract (8.1) 21.5 3,741 Thick motherwort extract (8.1) 43.5 7,561 Taurine (8.2) 433.5 75.39 Microcrystalline cellulose (8.2) 35 6.08 Croscarmellose sodium (8.3) 5.5 0.96 Silicon Colloidal Dioxide (8.3) 5.5 0.96 Magnesium stearate (8.3) 5.5 0.96 Film-forming mixture: (8.4) 25 4.35 total weight 575 100.00

Coating of the tablets may be carried out in machines with a perforated boiler, a boiler with a chimney, or in fluidized bed apparatus.

Example 9

The method of obtaining the mass for filling capsules differs from that described in Example 6 in that the hydration and granulation of a mixture of taurine (9.2) with microcrystalline cellulose (9.2) is carried out with liquid plant extracts (50% or more by weight) (9.1), and also in that the dried granulate is mixed with the components (9.3). The resulting mass is Packed in hard gelatin capsules. This method is characterized by the simultaneous process of wet granulation and drying using a fluidized bed granulator dryer.

The composition of the dosage form in capsules:

Component No. Composition per 1 capsule (mg) % Liquid hawthorn extract (9.1) 43 3.91 Motherwort extract liquid (9.1) 87 7.91 Taurine (9.2) 867 78.82 Microcrystalline cellulose (9.2) 81 7.36 Silicon Colloidal Dioxide (9.3) eleven 1.00 Magnesium stearate (9.3) eleven 1.00 total weight 1100 100.00

Example 10

The method for producing tablets differs from that described in Example 6 in that a mixture of taurine, dry extracts of hawthorn and motherwort, crospovidone (components 10.1) is granulated dry using a compactor or briquetting, the obtained granulate is mixed with magnesium stearate (10.2) and pressed into tablets.

The composition of the dosage form in tablets:

Component No. Composition per 1 tablet (mg) % Dry hawthorn extract (10.1) 43 3.91 Dry motherwort extract (10.1) 87 7.91 Taurine (10.1) 867 78.82 Crospovidone (10.1) 92 8.36 Magnesium stearate (10.2) eleven 1.00 total weight 1100 100.00

Example 11

This method differs from those described in Examples 6-8 and 10 in that the mass obtained above is not compressed into tablets, but filled with hard gelatin capsules.

Example 12

The method of preparation of the dosage form consists in the fact that the dry complex obtained by evaporation of the thick extracts of motherwort, hawthorn and excipients obtained according to one of the following Examples 1-5, is mixed with taurine in the calculated ratio, the mixture is compacted and hard gelatin capsules are filled with granulate.

The composition of the dosage form in capsules:

Component No. Composition per 1 capsule (mg) % Complex extract of hawthorn and motherwort dry (Examples 1-5) (12.1) 193 18.21 Taurine (12.2) 867 81.79 total weight 1060 100.00

Example 13

It differs from Example 12 in that the dry complex is mixed with taurine and other auxiliary components and pressed into tablets.

The composition of the dosage form in tablets:

Component No. Composition per 1 capsule (mg) % Complex extract of hawthorn and motherwort dry (Examples 1-5) (13.1) 193 17.54 Taurine (13.1) 867 78.82 Crospovidone (13.1) 29th 2.64 Magnesium stearate (13.2) eleven 1.00 total weight 1100 100.00

Preclinical studies of the pharmaceutical composition of the above composition were aimed at studying its influence on the main risk factors for cardiovascular disease in the conditions of metabolic syndrome (prediabetes) and diabetes mellitus. In order to establish the mechanisms of the cardioprotective effect of the drug, its effect on bioenergetic processes and the oxidative status of myocardial mitochondria in animals with metabolic syndrome was studied.

The theoretical justification for the prospects of using a pharmaceutical composition containing taurine, extracts of hawthorn and motherwort for the prevention and treatment of metabolic syndrome and diabetic nephropathy was established pharmacotherapeutic properties of the above components.

When studying the effect of taurine on many parts of the pathogenesis of insulin resistance, premature atherosclerosis and diabetic nephropathy, a positive corrective effect of this amino sulfonic acid was shown [1-3]. Taurine was found to improve the sensitivity of peripheral tissues to insulin and reduce abdominal obesity in rats with type 2 diabetes mellitus [1], reduce the concentration of triglycerides in the blood serum of rats with streptozotocin diabetes [4], and increase the level of reduced glutathione in isolated hepatocytes [5] ]. It was shown that the consumption of high doses of taurine prevents the development of functional and structural disorders of the kidneys in animals with streptozocin diabetes, reducing the degree of proteinuria, reducing glomerular hypertrophy and the development of glomerulosclerosis [6]. From literary sources it is known that taurine exhibits a pronounced pharmacological effect on the cardiovascular system. By regulating the functioning of the renin-angiotensinaldesterone and kallikrein-kinin systems, affecting the activity of calcium-dependent enzymes and enzyme systems that perform catalytic functions in the plasma membranes of cardiomyocytes, sarcoplasmic reticulum of cardiomyocytes, and endoplasmic cardiomyocytoma of the brain cells, action [7, 8, 9].

Very important is the hypoglycemic effect of taurine, which has been confirmed in the medicine Dibikor, used for type 2 diabetes mellitus [10, 11]. The pharmacological effects of taurine indicate its specific modulating effect against many biochemical and physiological processes of the cell under normal and pathological conditions.

Given the pharmacological properties of motherwort and hawthorn extracts, due to the high antioxidant activity and the ability to regulate bioenergy processes in myocardial cells, they were expected to show a significant pronounced cardioprotective effect in the pharmaceutical composition.

Earlier in the experiment, it was shown that the antiatherosclerotic effect of hawthorn and motherwort extracts is caused by a complex of pharmacologically active components - flavonoids, phenolic acids, triterpenes, catechins, saponins and cyanidin derivatives [12, 13], i.e. substances with high antioxidant potential. This was confirmed by the fact that cardiotonic medicines of hawthorn exhibit a rhythmic effect, causing a positive isotropic and negative chronotropic effect on the myocardium. This therapeutic effect is manifested in an increase in the amplitude of heart contractions and a decrease in their frequency [14]. It was shown that the subsequent development of the hypotensive effect is directly related to the pharmacological properties of hawthorn extract saponins [15]. It should be noted that the pronounced hypocholesterolemic effect of hawthorn preparations is also associated with these substances [16].

Sedative properties of motherwort extract, depending on the dose, are 2-3 times higher than valerian preparations. This effect is associated with the inhibitory effect of motherwort preparations on the central nervous system, the regulatory effect on CCC with a mandatory decrease in the heart rate, with an increase in diastole followed by a decrease in blood pressure [15]. In the clinic of neurogenic cardiomyopathies uncomplicated by organic disorders, preparations of motherwort extracts showed a pronounced effect on the 5-6th day of their use [17].

The examples of pharmacological effects of taurine, extracts of hawthorn and motherwort indicate the promise of using the claimed pharmaceutical composition for the prevention and treatment of metabolic syndrome and diabetic nephropathy.

In the method for the prevention and treatment of metabolic syndrome (MS) and diabetic nephropathy, the pharmaceutical composition X (FC X) is used in the form of a granulate obtained in Example 6.

The method was tested on male Wistar rats weighing 290-300 g. The effect of FC X on the development of metabolic syndrome and diabetic nephropathy was studied using experimental models of the metabolic syndrome and diabetes mellitus, which are as close as possible to the corresponding pathologies in humans.

Metabolic syndrome induced by a high-fat diet.

Metabolic syndrome was induced by 8 weekly animals on a high-fat diet (VZD), in which proteins, carbohydrates, fats were 4.9; 26.4; 68.8%, respectively, of total calories [18].

FC X was administered orally with a probe in the form of an aqueous suspension in doses of 100 or 300 mg of taurine per kg of body weight for 8 weeks starting from the first day of the experiment.

Metabolic syndrome induced by a high fructose diet.

The model of metabolic syndrome was induced in rats by chronic (within two months) intake of fructose with drinking water at a concentration of 200 g / l [19].

FC X was administered orally with a probe in the form of an aqueous suspension for 8 weeks, starting from the first day of the experiment at a dose of 100 mg of taurine per kg of body weight.

Streptozotocin diabetes model.

Diabetes was caused by a single intraperitoneal injection of streptozotocin at a dose of 60 mg / kg body weight [20]. One week after the induction of diabetes, animals with basal glycemia of more than 14 mmol / L were included in the experiment. FC X was administered orally with a probe in the form of an aqueous suspension in doses of 100 and 300 mg of taurine per kg of body weight from the first day of the experiment for 8 weeks.

Control groups received a placebo in a similar pattern (placebo composition: potato starch; magnesium stearate; aerosil).

An oil solution of vitamin E (Tocopherol acetate, Slovakofarma company) at a dose of 50 mg / kg body weight was used as a comparison drug.

At the end of the experiment, the state of glucose homeostasis was assessed by basal glycemia, glucosuria, during an intraperitoneal glucose tolerance test (WBTG) (3 g / kg body weight) and by the level of fructosamine in blood serum [21]. Insulin sensitivity was determined using an intraperitoneal test of insulin tolerance (WBTI) (insulin 0.5 U / kg, glucose 2 g / kg body weight 10 min after insulin administration) [22] and a short insulin test (0.5 U / kg body weight) [23]. Visceral obesity was characterized by the mass of visceral fat, which was calculated as the sum of epigonadal, epinephral and mesenteric fat.

The level of non-esterified fatty acids (NEFA) in blood serum and liver homogenate was evaluated by the photocolorimetric method [24]. The level of total cholesterol and triglycerides in blood serum and liver and heart homogenates was determined by spectrophotometric methods [25, 26]. Organ biochemical parameters were expressed per mg of protein, which was evaluated by the Brad Ford method [27].

The intensity of oxidative stress was characterized by the concentration of cerulloplasmin [28] and diene conjugates (DC) [29] in blood serum, in the homogenate of the kidneys and heart, as well as by the content of malondialdehyde (MDA) [30] in the homogenate of the kidneys and liver. The state of the antioxidant defense system in the liver and heart was evaluated by the content of reduced glutathione (HB) [31], as well as by the activity of superoxide dismutase (SOD) [32]. The level of stable serum nitric oxide (NOx) metabolites was determined using the Griss reagent [33].

The state of the blood coagulation system was determined by the degree and speed of ADP-induced platelet aggregation [34]. Rat mitochondria were obtained by differential centrifugation. Mitochondrial respiration was determined by the polarographic method using a Clark closed oxygen electrode [35].

The effect of the drug on the state of the antioxidant defense of the mitochondrial system was evaluated by the content of reduced glutathione [31] in a suspension of isolated mitochondria. The rate of free radical oxidation in cardiac mitochondria was estimated by the level of lipid hydroperoxides [36]. When analyzing electrocardiograms (ECG) in three standard leads from the extremities, the following indicators were taken into account: the amplitude of the R, P, S, and T waves, heart rate (HR), the duration of the R-R, T-Q, Q-T intervals, and the QRS complex [37].

In order to verify microvascular pathology in the kidneys of rats with streptozotocin diabetes, a histological examination was performed using electron microscopy [38]. The thickness of the glomerular basement membrane was determined by the method of Hirose et al. [39].

The functional state of the kidneys was determined by daily urine output, urine pH, creatinine [40] and urea [41] in urine and blood serum, as well as by the level of microalbuminuria, which was determined using the Granum ELISA kits.

Statistical analysis of the results was carried out by methods of variation statistics. To determine the nature of the distribution of the results of the study, the Shapiro-Wilk test was used. In the case of a normal distribution, ANOVA analysis was performed, and the Nyumen – Cales test was used for multiple comparisons [42].

The effect of FC X on the development of a metabolic syndrome induced by a high-fat diet in rats.

As a result of the studies, it was found that the use of FC X significantly inhibits the development of insulin resistance and carbohydrate intolerance in rats with UR induced by MS, which was confirmed by a significant increase in the sensitivity coefficient to insulin and a decrease in the area under glycemic curves compared to the indices for the group, which received a placebo (Table 1). It should be noted that the effect of FC X on glucose homeostasis was not inferior to the comparison drug - vitamin E and did not increase with an increase in the dose to 300 mg / kg body weight.

It was found that the use of FC X both at a dose of 100 mg / kg and 300 mg / kg body weight, in contrast to vitamin E, led to a significant decrease in the relative mass of visceral fat by more than 30% and prevented the increase in body weight of rats compared to control animals. Moreover, the use of FC X in a larger dose was accompanied by a more pronounced effect on the relative mass of mesenteric fat (Table 2).

Table 1
Effect of FC X on glucose homeostasis in rats with metabolic syndrome induced by a high-fat diet (X ± Sx, n = 6) Group Basal glycemia, mmol / l The area under the glycemic curve, mmol / l · min The coefficient of sensitivity to insulin,% Intact control 5.14 ± 0.51 852.4 ± 48.2 39.4 ± 3.6 WZD + placebo 5.37 ± 0.36 1365.1 ± 89.4 ¹ 14.0 ± 1.84 ¹ VZD + FC X (100 mg / kg) 5.60 ± 0.47 994.6 ± 36.8 ² 36.5 ± 1.7 ² VZD + FC X (300 mg / kg) 5.04 ± 0.45 893.0 ± 15.9 ² 35.1 ± 2.7 ² VZD + Vitamin E 5.22 ± 0.15 922.0 ± 17.9 ² 40.1 ± 3.7 ² Notes: ¹ - Statistically significant differences in comparison with indicators for the "intact control" group, p <0.05; ² - Statistically significant differences in comparison with indicators for the group "VZHD + placebo", p <0.05; ³ - Statistically significant differences in comparison with indicators for the group "VZHD + FC X (100 mg / kg body weight)", p <0.05; ⁴ - Statistically significant differences in comparison with indicators for the group "VZhD + Vitamin E", p <0.05.

table 2 Effect of FC X on body weight gain and relative visceral fat mass in rats with metabolic syndrome induced by a high-fat diet (X ± Sx, n = 6) Group Weight gain,% The relative mass of visceral fat,% Intact control 42.01 ± 2.71 2.33 ± 0.24 WZD + placebo 69.12 ± 4.10 ¹ 6.28 ± 0.78 ¹ VZD + FC X (100 mg / kg) 41.83 ± 5.51 ^2.4 4.48 ± 0.43 ^1.2.4 VZD + FC X (300 mg / kg) 46.82 ± 5.33 ^2.4 3.84 ± 0.18 ^1.2.4 VZD + Vitamin E 66.23 ± 4.10 ¹ 6.13 ± 0.81 ¹

Note. Reliability indicators are similar to table 1.

As a result of the studies, it was found that the use of FC X in both doses, unlike vitamin E, helps to normalize triglyceridemia (Table 3). The hypolipidemic effect of pharmaceutical composition X also manifested itself in a decrease in the level of total cholesterol in the blood serum, even compared with the intact control.

Table 3 The effect of FC X on lipid metabolism in rat blood serum with metabolic syndrome induced by a high-fat diet (X ± Sx, n = 6) Group Triglycerides, mmol / L Total cholesterol, mmol / l Intact control 0.77 ± 0.04 2.35 ± 0.20 WZD + placebo 1.54 ± 0.20 ¹ 2.01 ± 0.16 VZD + FC X (100 mg / kg) 0.89 ± 0.07 ^2.4 1.61 ± 0.12 ^1.4 VZD + FC X (300 mg / kg) 0.91 ± 0.022 1.57 ± 0.13 ^1.4 VZD + Vitamin E 1.41 ± 0.19 ¹ 2.21 ± 0.11 Note. Reliability indicators are similar to table 1.

The use of FC X in both doses led to a decrease in the level of unesterified fatty acids in the homogenate of rat liver with MS. At the same time, normalization of triglycerides in this organ was observed only when using FC X at a dose of 300 mg / kg (Table 4).

Table 4
Effect of FC X on lipid metabolism in the liver and heart in rats with metabolic syndrome induced by a high-fat diet (X ± Sx, n = 6) Group Liver Heart Triglycerides, nmol / mg protein SFA, nmol / mg protein Triglycerides, nmol / mg protein SFA, nmol / mg protein Intact control 34.68 ± 4.27 89.87 ± 3.01 16.46 ± 0.64 51.74 ± 2.51 WZD + placebo 102.88 ± 17.65 ¹ 128.61 ± 8.02 ¹ 30.48 ± 2.16 ¹ 90.01 ± 3.67 ¹ VZD + FC X (100 mg / kg) 94.33 ± 14.74 ¹ 83.20 ± 8.00 ^2.4 25.84 ± 3.60 ¹ 59.93 ± 7.27 ^2.4 VZD + FC X (300 mg / kg) 44,21 ± 8,13 ^2,3,4 93.14 ± 10.72 ^2.4 14.94 ± 0.75 ^2.3.4 63.55 ± 7.79 ^2.4 VZD + Vitamin E 98.28 ± 13.55 ¹ 121.44 ± 9.22 ¹ 26.54 ± 3.86 ¹ 84.91 ± 4.66 ¹ Note. Reliability indicators are similar to table 1.

It was found that the level of triglycerides and SFA was significantly increased in the heart of rats with metabolic syndrome, while the administration of FC X at a dose of 100 mg / kg of body weight normalized the level of SFA and at a dose of 300 mg / kg, it was SFA and triglycerides (see table. four).

It should be noted that the effect of FC X on the lipid profile significantly exceeded the effect of the vitamin E comparison drug.

It is known that there is a relationship between endothelial dysfunction and insulin resistance, as evidenced by the deterioration of insulin sensitivity in conditions of reduced NO synthase activity [43]. In biological media, the level of stable decomposition products of NO — nitrites and nitrates — is defined as an indicator of NO production in vivo [44].

As a result of the studies, it was found that the concentration of stable NO metabolites in the blood serum and urine of rats contained on VZhD is significantly reduced compared to the intact control (Table 5). At the same time, the use of FC X in both doses and vitamin E led to the normalization of this indicator (see table 5).

Table 5 Effect of FC X on the concentration of stable NO metabolites in blood serum and urine in rats with metabolic syndrome induced by a high-fat diet (X ± Sx, n = 6) Group Serum NOx, µmol / L NOx in urine, µmol / L Intact control 31.97 ± 3.51 63.70 ± 4.26 WZD + placebo 20.26 ± 1.90 ¹ 45.87 ± 3.41 ¹ VZD + FC X (100 mg / kg) 35.67 ± 1.74 ² 66.01 ± 5.69 ² VZD + FC X (300 mg / kg) 31.96 ± 1.51 ² 63.04 ± 4.81 ² VZD + Vitamin E 28.76 ± 2.31 ² 60.34 ± 2.91 ² Note. Reliability indicators are similar to table 1.

A large amount of data has been accumulated indicating the relationship of an increased level of free radicals and insulin resistance, the main component of the metabolic syndrome, type 2 diabetes mellitus, and the leading risk factor for macrovascular pathology [45].

When determining the LPO intensity in the heart of rats with MS, a significant increase in the content of DC was found against a background of a decrease in the content of reduced glutathione and a compensatory increase in the activity of SOD (Table 6). The introduction of FC X in both doses led to a decrease in the level of DC and normalization of SOD activity, while its use at a dose of 300 mg / kg also contributed to an increase in the content of reduced glutathione in the heart of rats with MS compared with the intact control (see table 6).

It should be noted that the effect of FC X on oxidative status indicators was not inferior to the comparison drug vitamin E.

Table 6 The effect of FC X on indicators of cardiac oxidative status in rats with a metabolic syndrome induced by a high-fat diet (X ± Sx, n = 6) Group DC, nmol / mg protein VG, nmol / mg protein SOD, cu / mg protein Intact control 0.38 ± 0.09 59.01 ± 2.98 26.71 ± 4.90 WZD + placebo 1.12 ± 0.11 ¹ 40.12 ± 2.84 ¹ 42.91 ± 7.12 ¹ VZD + FC X (100 mg / kg) 0.44 ± 0.08 ² 44.55 ± 3.34 ¹ 22.00 ± 1.56 ² VZD + FC X (300 mg / kg) 0.39 ± 0.06 ² 65,88 ± 7,86 ^2,3,4 21.25 ± 3.00 ² VZD + Vitamin E 0.43 ± 0.07 ² 55.44 ± 3.14 ² 24.50 ± 2.06 ² Note. Reliability indicators are similar to table 1.

It is known that MS is accompanied by increased production of PAI-1 and fibrinogen, which leads to a decrease in thrombolysis and fibrinolysis and a shift in the hemostatic balance towards a thrombotic state [46, 47].

It was shown that in rats with UR-induced MS, both the degree and rate of ADP-stimulated platelet aggregation increase (Table 7). Unlike vitamin E, the administration of FC X at a dose of 100 mg / kg body weight led to a decrease in the aggregation index, and at the maximum dose to a significant decrease in both indicators.

Table 7 Effect of FC X on the rate and index of platelet aggregation in the blood of rats with metabolic syndrome induced by a high-fat diet (X ± Sx, n = 6) Group Aggregation Index,% Aggregation rate, unit Intact control 29.17 ± 3.16 59.78 ± 4.72 WZD + placebo 47.00 ± 2.07 ¹ 90.40 ± 7.70 ¹ VZD + FC X (100 mg / kg) 32.17 ± 3.96 ^2.4 75.83 ± 7.50 VZD + FC X (300 mg / kg) 28.00 ± 3.51 ^2.4 57.81 ± 4.64 ^2.4 VZD + Vitamin E 43.45 ± 3.47 ¹ 95.32 ± 4.35 ¹ Note. Reliability indicators are similar to table 1.

Thus, the developed FC X at doses of 100 and 300 mg / kg body weight inhibits such manifestations of metabolic syndrome induced by VZhD in rats, such as insulin resistance, carbohydrate intolerance, abdominal obesity, hypertriglyceridemia, oxidative stress, endothelial dysfunction, prothrombotic state. It should be noted that a dose of 100 mg / kg of body weight is sufficient for the manifestation of these pharmacological effects and can be recommended for use in order to reduce cardiovascular disease and prevent type 2 diabetes in people at risk.

The effect of FC X on the development of the metabolic syndrome induced by the high fructose diet in rats.

It was found that the two-month use of FC X, like vitamin E, inhibits the development of insulin resistance and carbohydrate intolerance in rats with metabolic syndrome induced by VFD, which was confirmed by a significant decrease in the area under the glycemic curves during VBTTG and VBTTI compared with the parameters for the group, which received a placebo (table 8). A decrease in the level of fructosamine in the blood serum of animals treated with FC X may be due to both an improvement in glucose homeostasis and its ability to directly inhibit the processes of nonenzymatic glucose addition to proteins [48].

Table 8 The effect of FC X on glucose homeostasis in rats with a metabolic syndrome induced by a high fructose diet (X ± Sx, n = 6) Group Basal glycemia, mmol / l The concentration of fructosamine, mmol / l PPK during VBTTG, mmol / l · min PPK during VBTTI, mmol / l · min Intact control 4.69 ± 0.28 1.24 ± 0.11 683.1 ± 27.3 379.9 ± 34.2 VFD + placebo 4.61 ± 0.35 2.34 ± 0.09 * 1043.7 ± 82.5 * 547.1 ± 18.8 * WFD + FC X 4.26 ± 0.23 1.66 ± 0.09 * ^# 695.5 ± 18.2 ^# 426.7 ± 38.7 ^# VFD + Vitamin E 4.55 ± 0.43 1.83 ± 0.08 * ^# 755.5 ± 19.1 ^# 468.1 ± 12.7 ^# Notes: * - Statistically significant differences in comparison with indicators for the "intact control" group, p <0.05; ^# - Statistically significant differences in comparison with indicators for the “VFD + placebo” group, p <0.05; ** - Statistically significant differences in comparison with indicators for the “VFD + vitamin E” group, p <0.05.

The use of FC X inhibited the increase in body weight in animals with MS, in particular, due to a decrease in the relative mass of visceral fat (Table 9).

Table 9 Effect of FC X on body weight gain and relative visceral fat mass in rats with metabolic syndrome induced by a high-fructose diet (X ± Sx, n = 6) Group Weight gain,% The relative mass of visceral fat. % Intact control 12.24 ± 2.05 1.67 ± 0.19 VFD + placebo 21.19 ± 1.63 * 3.95 ± 0.31 * WFD + FC X 13.54 ± 2.87 ^{# **} 2.37 ± 0.27 * ^{# **} VFD + Vitamin E 20.88 ± 0.93 * 4.02 ± 0.21 * Note. Reliability indicators are similar to table 8.

The introduction of FC X, unlike vitamin E, helped to normalize triglyceridemia and reduce the concentration of total cholesterol in blood serum, even in comparison with the intact control (see table 10).

Table 10 Effect of FC X on the lipid profile and the level of stable NO metabolites in the blood serum of rats with metabolic syndrome induced by a high-fructose diet (X ± Sx, n = 6) Group The concentration of triglycerides, mmol / l Total cholesterol, mmol / l NO _2- + NO _3- , μmol / L Intact control 0.69 ± 0.04 1.67 ± 0.09 31.60 ± 2.51 VFD + placebo 1.32 ± 0.10 * 1.93 ± 0.07 * 25.37 ± 0.94 * WFD + FC X 0.69 ± 0.06 ^{# **} 1.40 ± 0.04 * ^{# **} 31.37 ± 1.94 ^# VFD + Vitamin E 1.26 ± 0.11 * 2.03 ± 0.06 * 33.47 ± 1.67 ^# Note. Reliability indicators are similar to table 8.

As a result of the studies, it was found that the concentration of stable NO metabolites in the blood serum of rats with metabolic syndrome is significantly reduced compared with indicators of intact control (table 10). At the same time, the use of FC X, unlike vitamin E, led to the normalization of this indicator, which may indicate the restoration of NO production under the influence of this drug (see table 10).

When determining the respiration indices of isolated cardiac mitochondria in the presence of glutamate and malate, a decrease in the rate of ADP-stimulated respiration (V3) and respiratory control coefficient in the mitochondria of rats with MS was found to be compared with intact control (Table 11). This indicates the separation of respiration and oxidative phosphorylation, which is one of the main markers of mitochondrial dysfunction.

Table 11 Effect of FC X on the respiratory rate (in the presence of malate and glutamate) of isolated mitochondria of rat heart with a metabolic syndrome induced by a high fructose diet (X ± Sx, n = 6) Group V ₄ natom O / min / mg protein V ₃ natom O / min / mg protein V3 / v4 Intact control 14.24 ± 1.20 82.60 ± 7.48 5.83 ± 0.35 VFD + placebo 14.44 ± 1.18 66.21 ± 5.00 * 4.63 ± 0.25 * WFD + FC X 14.51 ± 1.26 76.04 ± 3.97 ^# 5.38 ± 0.32 ^# Note. Reliability indicators are similar to table 8.

It should be noted that the use of FC X led to an increase in respiratory control due to normalization of the rate of ADP-stimulated respiration of cardiac mitochondria in metabolic state 3 in the presence of malate and glutamate compared with the placebo group (see Table 11).

The results obtained may indicate an improvement in the processes of oxidative phosphorylation at the level of complex I of the electron transport chain under the influence of this drug.

It is known that under conditions of mitochondrial dysfunction, the generation of reactive oxygen species significantly increases, which, in turn, leads to oxidative disturbances in mitochondria and apoptosis and cell necrosis. Thus, there is a vicious circle between mitochondrial dysfunction and oxidative stress, which can be broken by normalizing the oxidative status of mitochondria [49].

It was shown that the use of FC X contributes to the reduction of TBA-active products in the mitochondria of the heart and the normalization of reduced glutathione (see Table 12).

Table 12 Effect of FC X on the content of TBA-active products and reduced glutathione in isolated cardiac mitochondria in rats with a metabolic syndrome induced by a high-fructose diet (X ± Sx, n = 6) Indicator Intact control VFD + placebo WFD + FC X TBA-active products, nmol / mg protein 0.43 ± 0.07 0.88 ± 0.07 * 0.65 ± 0.06 * ^# VG, nmol / mg protein 3.84 ± 0.28 2.10 ± 0.27 * 3.67 ± 0.29 ^# Note. Reliability indicators are similar to table 8.

The results obtained indicate that FC X restores bioenergetic processes in the heart muscle of animals with metabolic syndrome, enhancing respiratory control and preventing the overproduction of ROS in the mitochondria of cardiomyocytes.

Assessment of the functional state of the cardiovascular system of rats with MS showed that the use of FC X leads to a prolongation of the RR ¹ interval and normalization of heart rate, which indicates the resumption of normal heart rhythm and the prevention of sinus tachycardia (see table 13).

Table 13 Effect of FC X on the duration of PQ, RR ¹ intervals and heart rate in rats with a metabolic syndrome induced by a high fructose diet (X ± Sx, n = 6) Group RR ¹ sec Heart rate, bpm P-Q, s Intact control 0.127 ± 0.001 470.9 ± 4.8 0.037 ± 0.001 VFD + placebo 0.119 ± 0.001 ¹⁾ 505.4 ± 6.6 ¹⁾ 0.044 ± 0.001 * WFD + FC X 0.134 ± 0.003 ^# 450.2 ± 11.2 ^# 0.037 ± 0.003 ^# Note. Reliability indicators are similar to table 8.

In addition, the administration of PK X prevented a reduction in the P-Q interval and contributed to the normalization of atrioventricular conduction in rats with MS (see Table 13).

In rats treated with FC X, normalization of the Q-T interval duration, prolongation of the TP interval and a decrease in the QT / TP ratio were observed (Table 14), which indicates the restoration of the processes of ventricular myocardial depolarization and repolarization.

Table 14 The effect of FC X on the duration of Q-T, TP and QT / TP ratios in rats with a high fructose diet-induced metabolic syndrome (X ± Sx, n = 6) Group Q-T, s TR, s QT / TP Intact control 0.074 ± 0.004 0.028 ± 0.002 2.81 ± 0.29 VFD + placebo 0.064 ± 0.003 ¹ 0.018 ± 0.002 * 3.83 ± 0.33 * WFD + FC X 0.075 ± 0.065 ^# 0.024 ± 0.001 ^# 3.28 ± 0.20 ^# Note. Reliability indicators are similar to table 8.

The results allow us to conclude that the normal heart rhythm and atrioventricular conduction resume, the systolic and diastolic activity of the heart improves, and sinus tachycardia in the myocardium is prevented under the influence of pharmaceutical composition X.

Thus, the application improves the functional state of the cardiovascular system due to the restoration of bioenergetic processes in the mitochondria of cardiomyocytes in animals with metabolic syndrome induced by VFD.

The effect of FC X on the development of diabetic rat nephropathy.

The risk of disability and mortality due to cardiovascular disease increases significantly in the presence of diabetic nephropathy, which is accompanied by an increase in blood pressure, a decrease in glomerular filtration and the development of inflammatory processes in the kidneys [50].

As a result of the experiment, it was found that the use of FC X at doses of 100 and 300 mg / kg body weight, unlike vitamin E, leads to a significant decrease in basal hyperglycemia by 30% and 50%, respectively, compared with the diabetic control (Table 15 )

A significant improvement in glycemic control under the influence of FC X was also evidenced by a significant decrease in the level of fructosamine in the blood serum of diabetic animals, which was noted when using the drug at a dose of 300 mg / kg body weight (see table 15).

Table 15 Effect of FC X on glucose homeostasis in rats with streptozotocin diabetes (X ± Sx n = 6) Group Basal glycemia, mmol / l Fructosamine, mmol / L Intact control 4.77 ± 0.13 1.83 ± 0.12 Diabetes + Placebo 15.72 ± 0.53 ¹ 3.19 ± 0.31 ¹ Diabetes + FC X (100 mg / kg) 11.28 ± 0.90 ^1.2.4 3.23 ± 0.22 ¹ Diabetes + FC X (300 mg / kg) 8.49 ± 1.84 ^1.2.4 2.48 ± 0.13 ^1.2.3 Diabetes + Vitamin E 14.47 ± 0.60 2.99 ± 0.12 Notes: ¹ - Statistically significant differences in comparison with indicators for the "intact control" group, p <0.05; ² - Statistically significant differences in comparison with indicators for the group "diabetes + placebo", p <0.05; ³ - Statistically significant differences in comparison with indicators for the group "diabetes + FC X (100 mg / kg)", p <0.05; ⁴ - Statistically significant differences in comparison with indicators for the group "diabetes + vitamin E", p <0.05.

The use of FC X in both doses, unlike vitamin E, reduced daily diuresis and glucosuria in diabetic animals, which correlated with an improvement in glycemic control under its influence (Table 16).

Table 16 Effect of FC X on the functional state of the kidneys and the thickness of the basement membrane of the vessels of the renal glomerulus in rats with streptozotocin diabetes (X ± Sx, n = 6) Indicator Daily diuresis, ml Glucose in urine, mmol / l The thickness of the basement membrane, microns Intact control 4.73 ± 0.50 - 0.197 ± 0.009 Diabetes + Placebo 19.02 ± 0.96 ¹ 134.52 ± 20.17 ¹ 0.357 ± 0.005 ¹ Diabetes + FC X (100 mg / kg) 16.10 ± 0.76 ^1.2 83,11 ± 13,08 ^1,2,4 0.203 ± 0.009 ^2.4 Diabetes + FC X (300 mg / kg) 13.22 ± 0.80 ^1.2.4 72,06 ± 22,34 ^1,2,4 0.201 ± 0.015 ^2.4 Diabetes + Vitamin E 19.89 ± 0.56 ¹ 122.21 ± 11.45 ¹ 0.364 ± 0.009 ¹ Note. Reliability indicators are similar to table 15.

An increase in the mesangial matrix and the thickness of the glomerular basement membrane is considered a characteristic morphological sign of diabetic nephropathy [51]. Using electron microscopy, it was shown that in the group of diabetic control the thickness of the glomerular basement membrane almost doubled that of intact animals (see Table 16), while in animals treated with FC X in both doses, this indicator was practically the same as intact control. It was found that the use of FC X in both doses prevents the development of functional impairment of the kidneys, as evidenced by the normalization of creatinine clearance and glomerular filtration rate in diabetic animals (see table 17).

One of the main pathogenetic mechanisms of diabetic complications is considered to be a violation of the redox balance - oxidative stress [52].

In animals treated with FC X in both doses, there was a significant decrease in the content of primary and secondary LPO products (DC and MDA) and an increase in the level of reduced glutathione in the kidneys compared with the diabetic control (Table 18), which indicates a pronounced antioxidant activity of the drug, not inferior in severity of the pharmacological effect to vitamin E.

Table 17 Effect of FC X on creatinine clearance and microalbuminuria in male rats with streptozotocin diabetes (X ± Sx, n = 6) Group Creatinine in urine, mmol / l Serum creatinine, mcmol / l Creatinine clearance, ml / min Microalbuminuria, mcg / l Intact control 17.38 ± 1.52 55.31 ± 3.16 1.02 ± 0.10 0.63 ± 0.13 Diabetes + Placebo 3.45 ± 0.20 ¹ 125.38 ± 4.26 ¹ 0.36 ± 0.04 ¹ 2.16 ± 0.34 ¹ Diabetes + FC X (100 mg / kg) 5.88 ± 0.61 ^1.2.4 68.83 ± 10.37 ^2.4 1.06 ± 0.19 ^2.4 0.78 ± 0.21 ^2.4 Diabetes + FC X (300 mg / kg) 6.43 ± 0.53 ^1.2.4 51.63 ± 6.32 ^2.4 1.27 ± 0.23 ^2.4 0.80 ± 0.11 ^2.4 Diabetes + Vitamin E 3.92 ± 0.72 ¹ 114.33 ± 7.45 0.40 ± 0.12 1.99 ± 0.14 Note. Reliability indicators are similar to table 15.

Table 18 The effect of FC X on the indicators of oxidative status in the kidneys of rats with streptozotocin diabetes (X ± Sx, n = 6) Show-
bodies Intact control Diabetes + Placebo Diabetes + PCH (100 mg / kg) Diabetes + PCH (300 mg / kg) Diabetes + Vitamin E DC, nmol / mg protein 0.16 ± 0.02 0.56 ± 0.04 ¹ 0.38 ± 0.06 ^1.2 0.30 ± 0.05 ^1.2 0.37 ± 0.05 ^1.2 MDA, nmol / mg protein 0.57 ± 0.04 1.51 ± 0.12 ¹ 0.87 ± 0.07 ² 0.85 ± 0.14 ² 0.77 ± 0.07 ² VG, nmol / mg protein 59.83 ± 4.93 32.67 ± 6.86 ¹ 90,04 ± 5,63 ^1,2,4 85.66 ± 6.86 ^1.2.4 55.14 ± 4.3 ² Note. Reliability indicators are similar to table 15.

The use of FC X, like vitamin E, in both doses normalized the level of nitric oxide metabolites in the blood serum and urine of diabetic animals (Table 19), which is probably due to its ability to suppress the development of oxidative stress in hyperglycemia.

Table 19 Effect of FC X on the level of nitric oxide metabolites in the urine and blood serum of rats with streptozotocin diabetes (X ± Sx, n = 6) Show-
bodies Intact control Diabetes + Placebo Diabetes + FC X (100 mg / kg) Diabetes + FC X (300 mg / kg) Diabetes + Vitamin E Serum NOx, µmol / L 33.79 ± 0.17 55.47 ± 4.71 ¹ 34.25 ± 2.32 ² 30.68 ± 2.42 ² 39.35 ± 2.12 ² NOx in urine, µmol / L 70.89 ± 4.49 100.68 ± 7.48 ¹ 72.60 ± 2.41 ² 71.92 ± 3.75 ² 79.56 ± 5.21 ² Note. Reliability indicators are similar to table 15.

Thus, the results obtained indicate that the use of FC X in doses of 100 and 300 mg / kg helps to reduce basal hyperglycemia, glucosuria, normalize creatinine clearance and glomerular filtration rate, reduce oxidative and nitrosative stress in the kidneys, as well as the severity of microalbuminuria in diabetic animals, which justifies the prospects of its use for the prevention and treatment of diabetic nephropathy. It should be noted that the effect of FC X at a dose of 100 mg / kg on most of the studied parameters was not inferior to its effect when used at a dose of 300 mg / kg body weight, which can be the basis for the use of the minimum dose in clinical practice.

FC X at a dose of 100 mg / kg body weight not only prevents the development of insulin resistance, carbohydrate intolerance, abdominal obesity, hypertriglyceridemia, hypercholesterolemia, oxidative stress, but also improves the functional state of the cardiovascular system due to the restoration of bioenergetic processes in animal mitochondria with cardiomas metabolic syndrome.

The revealed pharmacological properties of FC X substantiate the promise of its use for the prevention and treatment of metabolic syndrome, correction of mitochondrial bioenergetic processes of the cardiovascular system and diabetic nephropathy.

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Claims (10)

1. A pharmaceutical composition for the prevention and treatment of metabolic syndrome and diabetic nephropathy, containing active substances taurine, motherwort herb extract, hawthorn fruit extract and excipients, characterized in that the components are taken in the following ratio, wt.%:
Dry hawthorn fruit extract 2-5 Dry motherwort herb extract 6-10 Taurine 65-80 Excipients rest * - (in terms of dry matter) 2. The pharmaceutical composition according to claim 1, characterized in that the dry extracts of the fruits of hawthorn and motherwort grass are introduced into the composition in the form of a complex with excipients. 3. A tool for the prevention and treatment of metabolic syndrome and diabetic nephropathy, containing the active substances taurine, motherwort herb extract, hawthorn fruit extract and excipients, characterized in that it contains a pharmaceutical composition in the following ratio, wt.%:
Dry hawthorn fruit extract 2-5 Dry motherwort herb extract 6-10 Taurine 65-80 Excipients rest * - (in terms of dry matter) 4. The tool according to claim 3, characterized in that as excipients selected substances that perform the functions of binders, agents that cause disintegration, and materials that contribute to the sliding and lubrication of granules during pressing, and film coating materials. 5. The tool according to PP.3-4, characterized in that as substances that act as binders, contains starch potato or microcrystalline cellulose to obtain granules for tabletting or encapsulation. 6. The tool according to PP.3-4, characterized in that as agents that cause disintegration, contains croscarmellose sodium to obtain granules with dry plant extracts. 7. The tool according to PP.3-4, characterized in that as materials that contribute to the sliding of the granulate during pressing and film coating materials, contains colloidal silicon dioxide. 8. The tool according to claims 3 to 4, characterized in that, as materials that promote lubrication, it contains magnesium stearate, which is introduced into the dry complexes with plant extracts upon receipt of granules for tabletting and encapsulation. 9. The tool according to claims 3-8, characterized in that it contains components in the form of dosage forms of tablets or film-coated tablets or capsules at a daily dose of 100 mg per taurine per kg of body weight. 10. A method of obtaining funds for the prevention and treatment of metabolic syndrome and diabetic nephropathy, characterized in that previously dry extracts of hawthorn fruit and motherwort herb are obtained by extraction with an aqueous or aqueous ethanol solvent, maceration or percolation, filtration extraction, or re -colation followed by drying dry extract, after which they are mixed with taurine and excipients that act as binders, disintegrating agents , and materials that contribute to the sliding and lubrication of the granulate during pressing.