EA001099B1 - Injecting medical preparation "citoflavin" possessing cito-pretecting effect - Google Patents

Abstract

An injecting medical preparation, having cito-protecting effect, comprising an active composition of natural metabolites and coenzymes in a solvent, characterized in that comprises a succinic acid and riboxyne (inosin) as natural metabolites, and nicotinoamide and riboflavin-5-sodium mononucleotide sodium as coenzymes, and water for injecting as a solvent, and further comprises sodium hydroxide and N-methylglycamine, having the following ratio of components, wt. % : Succinic asid 9.5-10.5 Riboxyne (inosin) 1.9-2.1 Nicotinoamide sodium mononucleotide 0.19-0.21 Sodium hydro 3.3-3.7 N-methylglycamine 15.7-17.3 Water for injecting up to 100

Description

The invention relates to medicine, in particular to a complex injectable drug with a cytoprotective effect.

The drug can be used to treat acute myocardial infarction, cerebral ischemia, hypoxic stroke, coronary insufficiency, coronary heart disease, liver and kidney diseases.

A large number of different pharmacological activities of succinic acid, its salts, succinates and derivatives, are known.

Among them, an antihypoxic agent based on succinic acid succinate for the treatment of fetal pathology of the fetus associated with hypoxia and infection (Russian patent 2056842, IPC A 61K 31/19, 06/30/92); hepatoprotective agent based on derivatives of succinic acid (Japanese patent 09/188.664, IPC C 07C 323/51, 07/22/97); a medicine for the treatment of cerebral ischemia based on succinic acid succinate (Russian patent 2108095, IPC A 61K 31/19, 11/22/94).

However, in the list of medicines approved for medical use and introduced into medical practice that contain succinic acid as the main active ingredient, the only dosage form is Limontar tablets, (0.2 g of free succinic acid and 0.05 g of citric acid) used in adults as a diagnostic tool to study the secretory ability of the stomach. (Mashkovsky MD Drugs. T.2 - Ed. 13th, new - Kharkov, “Torsing”, 1997, p. 494).

A wide range of pharmacological activity of succinic acid is due to the fact that it is a natural metabolite of the body, participating in the redox cycle of dicarboxylic acids (Krebs cycle) and thereby ensuring normal cell activity.

Also known are complex preparations based on natural purine nucleosides, for example, inosine (riboxin), guanosine, adenosine, which are used as drugs of narrow action or cardioprotective (Russian patent 1769428, IPC A 61K 31/70, 09.04.90); (Russian patent 2035908, IPC A 61K 31/52, 09/24/91) or hepatoprotective (Japan patent 61,277619, IPC A 61K 3 1/505, 01/01/85).

Closest to the claimed is a comprehensive injectable drug with a narrowly targeted hepatoprotective effect, containing a mixture of metabolites and vitamins (UK patent 1178984, IPC A 61K 25/00, 06/23/66), the following composition:

Calcium salt of 1-b-5-formyl tetrahydrofolic acid 0.9 mg

Hydroxycobalamin hydrochloride 0.5 mg

Cyanobalamin 0.5 mg

Riboxin 60 mg

Adenine 20 mg

Nicotinamide 10 mg

Riboflavin-5-mononucleotide sodium 2.73 mg

The rest is distilled sterile- Up to 3 g naya water for injection

However, with severe pathologies, there is a need for complex medications that protect the affected cell and at the same time normalize its vital activity.

With various pathological processes in the body, the normal vital activity of cells is disrupted, which is manifested in a deterioration in the respiratory (redox) function of the cells, a decrease or termination of the synthesis of proteins and enzymes by the cell, and other disorders. Ultimately, these disorders lead to damage to cell membranes, a decrease in the level of metabolite utilization, and cell death.

Therefore, the creation of complex drugs that protect the cell from destruction and death and contribute to its normal functioning, the so-called cytoprotectors, is an urgent and important task.

Such drugs should contain both substances that enhance the metabolic processes, and components that contribute to the rapid and effective utilization of metabolites in the cells of damaged organs and tissues.

In this regard, the applicant has developed a complex drug with a cytoprotective effect, which includes both metabolite components (succinic acid and riboxin) and cofermet components - vitamins (nicotinamide and riboflavin-5-mononucleotide sodium), in the following ratio of components, wt.%:

Succinic acid 9.5-10.5

Riboxin (Inosine) 1.9-2.1

Nicotinamide 0.95-1.05

Riboflavin-5-mononucleotide sodium 0.19-0.21

Sodium hydroxide 3.3-3.7

Ν-Methylglucamine 15.7-17.3

Water for injection Until 1 00

An example of a specific preparation of "Cytoflavin" for injection

50.0 l of water for injection, 10.0 kg of succinic acid, 3.4 kg of sodium hydroxide, 0.6 kg of riboflavin-5-sodium mononucleotide, 16.5 kg of methyl glucamine, 1 are loaded into a glass apparatus with a 100-liter stirrer. , 0 kg nicotinamide, 2.0 kg riboxin. The resulting mixture is stirred until the components are completely dissolved and then water for injection is added until a volume of 100 l is obtained. The resulting solution is filtered through a Pall type sterilizing filter, poured into 5 ml sterile ampoules and sealed. Receive 18,000 ampoules with a volume of 5 ml. Yield 90%.

Each ampule contains - succinic acid 500 mg (10%), riboxin 100 mg (2%), nicotinamide 50 mg (1%), riboflavin-5 sodium mononucleotide 10 mg (0.2%), sodium hydroxide 170 mg (3.4 %), Ν-methylglucamine 825 mg (16.5%), water for injection up to 100%.

The use of this drug allowed to obtain a high cytoprotective effect on various models of cell damage in pathology of the heart, liver and kidneys.

The study of the cytoprotective activity of the drug

The study of the claimed drug was carried out in comparison with the prototype.

Experience 1. The study of cardioprotective action on the model of antiarrhythmic activity.

Given that arrhythmias are one of the first indicators of the development of myocardial cell pathology in various cardiovascular diseases accompanied by damage to myocardial cells, such as acute myocardial infarction and coronary heart disease, it was necessary to study and evaluate the cytoprotective effect of “Cytoflavin” on myocardial cells first of all, the antiarrhythmic activity of the proposed drug.

The experiment was carried out on linear white CBA mice, divided into five groups of 25 animals.

Experimental cardiac arrhythmia was modeled by the conventional method — by administering an alkonoid aconitine subcutaneously in a single dose of 200 μg / kg of animal weight and propranol subcutaneously in a single dose of 40 mg / kg.

The introduction of the alkaloid aconitine into the body causes a steady increase in sodium permeability of the membranes of cardiomyocytes (Orlov B.N. Poisonous animals and plants of the USSR. - M., Higher School, 1990, p. 192 - 194).

The introduction of propranol causes bradycardia and cardiac arrest due to blockade of β-adrenergic receptors in the myocardium and changes in the intracellular ratio of potassium and sodium ions. (Schottkde Ogoyr op Atbsh1a5-S1gsiayop. 1991, v. 84, No. 4. p. 1831 - 1851).

"Cytoflavin", a prototype and control 0.9% isotonic sodium chloride solution was injected intravenously into the tail vein at a single dose of 1.2 ml / kg of animal weight, once five minutes before administration of aconitine or propranol. Recording of the electrocardiogram in mice was carried out at the beginning of the experiment, and then 10, 30 and 60 minutes after drug administration.

To determine the time of occurrence and duration of extrasystole during the experiment, an electrocardiogram was monitored for 60 minutes.

The obtained experimental data were statistically processed using Student's criterion at P <0.05.

Data on testing the antiarrhythmic activity of the drugs are presented in table 1.

When simulating aconitine arrhythmia, administration of aconitine caused a polytonic extrasystole in 1 00% of the animals (see Table 1).

With the introduction of Cytoflavin at a dose of 1.2 ml / kg 5 minutes before administration of aconitine, extrasystole developed in only 25% of mice, while the latent period of extrasystole decreased by 35%, and the number of extrasystoles itself decreased by 2.4 times.

Thus, “Cytoflavin” showed a pronounced cytoprotective antiarrhythmic effect on myocardial cells in the model of aconitine arrhythmia. Given the mechanism of action of aconitine alkaloid, the mechanism of the protective effect of "Cytoflavin" is due to stabilization of the cell membrane of cardiomyocytes and a decrease in their permeability to sodium ions.

On the contrary, with the introduction of the prototype solution 5 minutes before the administration of aconitine, the opposite picture developed: extrasystole was observed in 75% of mice, and the latent period, duration and total number of extrasystoles did not change at all compared to aconitine.

Thus, the prototype showed a mild protective antiarrhythmic activity on myocardial cells in the model of aconitine arrhythmia and does not have a stabilizing effect on cardiomyocyte cells in this model.

In arrhythmias caused by the administration of propranol, polytonic extrasystole was also caused in 1 00% of mice.

With the preliminary administration of “Cytoflavin” at a dose of 1.2 ml / kg into the tail vein five minutes before the administration of propranol, extrasystole developed in 40% of the animals, and the latent period of the extrasystole decreased by 29%, and the total number of extrasystoles decreased by 3.2 times. The data obtained indicate a high cytoprotective effect of “Cytoflavin” on myocardial cells in a model of arrhythmia caused by the introduction of propranol. The protective mechanism is due to the normalization of the biochemistry of sodium / potassium metabolism through the cell membrane.

With the introduction of the prototype solution, the polytonic extasystole was detected in 50% of the mice, the latent period of the extrasystole practically did not change, and the average number of extrasystoles decreased only 1.7 times (Table 1.). This indicates a moderate protective effect of the prototype on myocardial cells.

Thus, on the model of propranol arrhythmia, Cytoflavin also exhibits a pronounced cytoprotective antiarrhythmic effect, and the prototype solution has only a weak protective antiarrhythmic effect.

This difference is explained by the creation of an optimal combination of natural metabolites - succinic acid (energy component), riboxin (substrate for ATP synthesis) and coenzymes-vitamins to obtain a cytoprotective effect. Due to the acceleration of the metabolism assimilation by the body’s cells, the biochemical processes inside the cell are accelerated and the protective effect on the myocardial cells, cardiomyocytes, occurs.

Experience 2. The study of cardioprotective action on the model of myocardial ischemia.

Ischemia (lack of oxygen) in myocardial cells leads to disruption of biochemical processes inside cardiomyocytes associated with energy consumption, followed by necrosis (destruction) of the cell membrane.

The study of the anti-ischemic activity of "Cytoflavin" in comparison with the prototype solution was carried out on white linear rats of the Wistar breed. All animals were divided into three groups of 25 animals.

For the study, we used the generally accepted model of ischemia caused by occlusion (surgical reduction of the vessel diameter - ligation) of the left coronary artery (\ νίη51ο \ ν E. Mešobk ίοτ 1Нс беУсбоппб акеккieпт! Οί арОггНубишссюубу. / Рогутупо. p. 401-433).

This model best illustrates the development of acute myocardial infarction in humans.

Rats were anesthetized with intraperitoneal administration of sodium etaminal at a dose of 50 mg / kg, the electrocardiogram was recorded and recorded.

Rats with rhythm disturbances were withdrawn from the experiment. A total of 36 rats were selected, divided into three groups of 1 2 animals.

In the remaining animals, the femoral vein and femoral artery were catheterized. Then, with mechanical ventilation of the lungs, the rats were opened in rats and the descending branch of the left coronary artery was ligated at the lower edge of the left atrial ear. All animals recorded mean arterial blood pressure in the femoral artery using a mercury manometer, and an electrocardiogram was monitored for 30 minutes to detect early ventricular arrhythmias developing against myocardial ischemia.

In the first group of animals, Cytoflavin was injected into the tail vein at a dose of 1.2 ml / kg five minutes after ligation.

In the second group, a solution of the prototype was administered at a dose of 1.2 ml / kg.

In the third control group, 0.9% physiological sodium chloride solution was injected into the tail vein at a dose of 1.2 ml / kg.

Throughout the experiment, artificial respiration was continued to prevent respiratory arrest caused by postoperative pneumothorax. According to the electrocardiogram, heart rate, mean blood pressure, incidence and total duration of periods of ventricular fibrillation and ventricular tachycardia were evaluated.

The effectiveness of the drugs according to the general protective cytoprotective effect on body cells was evaluated by mortality in each group.

Throughout the experiment, blood pressure in the femoral vein was recorded in all animals and the electrocardiogram was recorded 1, 5, 15, 30, 45, and 60 min from the beginning of the introduction of solutions into the tail vein.

Statistical significance was evaluated using Student's criterion. Generalized experimental data are presented in table. 2.

The results of the study of the anti-ischemic activity of “Cytoflavin” showed that with ischemia of the heart, the drug significantly reduces 2.62 times the number of ventricular extrasystoles in comparison with the control and 1.6 times in comparison with the prototype.

The duration of tachycardia decreased by 3 times compared with the control and 2.1 times compared with the prototype, although the incidence of tachycardia did not decrease in all cases. “Cytoflavin” also significantly reduced the frequency of ventricular fibrillation by 2.2 times compared with the control and 1.5 times compared with the prototype, although it had practically no effect on the duration of fibrillation time. The data obtained indicate a high cytoprotective activity of "Cytoflavin" on myocardial cells under ischemic conditions.

Data on the survival of animals under experimental conditions also indicate a high cytoprotective activity of Cytoflavin, with the introduction of which 1 00% (1 2 rats) survived after ligation, with the introduction of the prototype - 50% (6 rats), and with the introduction of a physiological saline - only 25% (3 rats) (see tab. 3).

Monitoring of mean arterial pressure and heart rate (see Table 4) showed that Cytoflavin faster than the prototype restored mean arterial pressure in the early period after occlusion, with practically no effect on heart rate. This effect indicates a more pronounced compensatory effect of "Cytoflavin" in comparison with the control and prototype in the restoration of biochemical processes in the cells during ischemia of cardiomyocytes, and therefore, the general cytoprotective effect on the myocardium.

Experience 3. The study of hepatoprotective and renoprotective activity of "Cytoflavin" in case of ethylene glycol poisoning.

To model lesions of liver and kidney cells, a well-known model of toxic hepatitis and nephroso-nephritis poisoning with 1,2-ethylene glycol was used. (Reproduction of diseases in animals for experimental and therapeutic research. Ed.

H. V. Lazareva. M .: "Medgiz", 1964, 392 p.).

In the experiment, 6 experimental groups of animals of 1 to 2 rats were formed: A-intact animals that were injected with saline; B - intact animals, which were introduced "Cytoflavin"; In - intact animals that were introduced the prototype; G-animals with ethylene glycol intoxication without treatment; D - animals with ethylene glycol intoxication and treatment with "Cytoflavin"; E - animals with ethylene glycol intoxication and prototype treatment.

Ethylene glycol was administered once intragastrically through an atraumatic probe in a dose

I, 5 g / kg. "Cytoflavin", the prototype and physiological saline in the control were injected into the tail vein of rats once a day for five days at a dose of 1.2 ml / kg. Biochemical studies were carried out according to standard methods (Steel I.D., Garishvili T.G. Modern methods in biochemistry. M: Medicine, 1978, 95 p. Clinical evaluation of laboratory tests / Edited by N.U. Titsa, M. : Medicine, 1986, 480 p.). The obtained experimental data were processed by statistical methods according to Student Fisher.

The clinical picture in the group of rats receiving ethylene glycol without treatment was characterized by inactivity, inhibition of reactions, ruffled coat and untidiness of animals. On the 2nd day, they developed macrohematuria, and from the 4th day oliguria. Biochemical and histological analysis of the liver and kidneys confirmed the development of damage to the kidneys and liver - nephrosonephritis and toxic hepatitis. Ethylene glycol intoxication disrupted all liver functions: protein-synthesizing and pigmented, which was accompanied by biochemical signs of cytolysis.

All animals, without exception, developed pronounced symptoms of kidney damage: in the analysis of urine, red blood cells, protein, cylinders, oxalic acid crystals, and kidney cells were determined. All kidney functions suffered - excretory, extretory and secretory, and in the blood products of nitrogen metabolism and cytolysis indicators increased.

During a morphological examination in the kidneys, degenerative changes and minor hemorrhages in the organ parenchyma were detected, fatty degeneration of the neuroepithelium was observed, small drops of fat were visible in the cytoplasm of the convoluted tubule cells.

Compared with the prototype, the hepatoprotective efficacy of Cytoflavin was judged by the clinical picture, the content of total bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase, total protein, serum lipids, glycogen, liver glutathione and test hexene .

Experimental data on the hepatoprotective effect of "Cytoflavin" are presented in table 5.

Studies have shown that the introduction of "Cytoflavin" and a prototype in healthy animals (groups A, B, C) does not cause liver dysfunction and behavioral reactions in rats.

The introduction of ethylene glycol (group D) was accompanied by a serious violation of all liver functions.

A decrease in total protein and lipids and glycogen stores in the liver indicates a violation of the protein synthesizing function of the liver.

The increase in the level of aminotransferases, alkaline phosphatase, bilirubin and thymol test indicates a significant destruction of liver cells - hepatocytes and a violation of pigment function.

An increase in the time of hexenal sleep, a decrease in the level of reduced glutathione and creatine phosphate shows a decrease in the detoxifying function of the liver.

The use of “Cytoflavin” for the treatment of liver damage caused by ethylene glycol (group D) showed reliable normalization of all liver functions: protein synthesizing (total protein, lipids, glycogen) to 8091% of normal, pigment (bilirubin, aminotransferase, alkaline phosphatase, thymol test) to 62-97% of the norm detoxifying (hexenal sleep, glutathione, creatine phosphate) to a level of 82-97% of the norm (group A).

Prototype treatment (group E) also had a positive effect after ethylene glycol poisoning, however, the prototype restored liver protein synthesis function by 60-90%, pigmented by 68-71%, and detoxified by only 50-70%.

Thus, “Cytoflavin” has a pronounced cytoprotective effect on liver cells - hepatocytes in case of ethylene glycol poisoning (see table 5).

The effectiveness of “Cytoflavin” as a renoprotector (protection of kidney cells) was evaluated by a set of biochemical parameters in comparison with the norm, control group and prototype: relative weight of the kidneys, daily urine output, urine protein, urine chlorides, urine sugar, urine density, urea, creatinine, lactate dehydrogenase.

Experimental data on the renoprotective effect of "Cytoflavin" are presented in table 6.

Experimental data showed that the introduction of both Cytoflavin and the prototype to healthy animals (groups A, B, C) did not adversely affect the functional activity of the kidneys.

Under conditions of pathology - ethylene glycol poisoning (group D), the weight of the kidneys increased in rats, which indicates edema. A significant increase in urine in the content of protein, chlorides, sugar, urea and an increase in urine density indicates a violation of the extretory and secretory function of the kidneys. A sharp decrease in the daily volume of urine amid an increase in creatinine and the enzyme lactate dehydrogenase indicates the destruction of renal tubule cells, the development of nephrosonephritis and, as a result, anuria.

The use of “Cytoflavin” (group D) for the treatment of kidney damage caused by ethylene glycol showed restoration of functional activity of the kidneys in all respects: edema decreased sharply (kidney mass became 94% of normal), excretory function (diuresis, urine density, creatinine) was restored to 62-100% secretory function (urine protein, sugar, chloride, urea, lactate dehydrogenase level) improved to 6282% of normal.

The use of the prototype (group E) for treatment also had a positive therapeutic effect in nephrosis-nephritis, but according to various indicators, the prototype was less effective than Cytoflavin: kidney mass was 73% of normal, excretory function was restored 51-93% and secretory function by 45-60% of the norm (see table. 6).

Analysis of histological data in animals of groups D and E also showed a sharp decrease in dystrophic changes in the tissue of the kidneys and liver, more pronounced in group D during treatment with "Cytoflavin".

Thus, cytoflavin has a pronounced cytoprotective effect on cells of the nephroepithelium and hepatocytes, superior to the prototype in experimental hepatitis and nephrosonephritis caused by the use of ethylene glycol.

Thus, a study of the biological activity of the drug showed that it has a cytoprotective effect, that is, it has a protective effect on the cells of the heart, liver and kidneys, normalizing the work of these organs in various pathologies.

To create a stable dosage form, the necessary quantities of Ν-methylglucamine and sodium hydroxide were experimentally selected. These components are not the active components of the dosage form, but provide the necessary physicochemical parameters.

Experience 4. The study of the influence of the content of г methylglucamine and sodium hydroxide on the stability of "Cytoflavin".

Considering the complex multicomponent composition of Cytoflavin and the presence of из-nucleoside - Riboxin (inosine) and Riboflavin mononucleotide easily hydrolyzed in aqueous solutions via an лик glycosidic bond in an acidic and alkaline environment (Biochemist Handbook: translated from English / Dawson R., Elliot D ., Jones K.M .: MIR, 1991, pp. 71-88, 115.), a study was made of the effect of the content of the stabilizer and complexing agent of Ν-methylglucamine on the pH of the solution and stability (absence of decomposition of components) during two-year storage at room temperature of aqueous races thief.

"Cytoflavin" is a drug intended for intravenous administration. The solutions introduced into the bloodstream must be physiologically compatible with the physicochemical parameters of the blood, in particular with pH. The introduction of acidic and / or alkaline solutions into the bloodstream is unacceptable, since this can lead to the formation of blood clots, damage to the walls of the veins, precipitation of drugs and the blockage of large and small blood vessels (I.A. Muravyov. -Technology of drugs, T. 2, Moscow, “Medicine”, 1980, S. 622-689, State Pharmacology of the USSR, M., Medicine, 1990 - S. 140).

We studied the effect of the amount of sodium hydroxide on the pH of the drug solution and the properties of the dosage form (table 8).

Studies have shown that the optimal content for this dosage form is the content of sodium hydroxide in an amount of 3.3 - 3.7%.

The experimental data of the experiment are presented in table 7.

Studies have shown that to create a stable drug, the content of methylglucamine in Cytoflavin is optimal from 15.7 to 17.3% - options D, D (Table 7) and sodium hydroxide content from 3.3 - 3.7% - options G, D (table 8).

Experience 5. Determination of acute toxicity of "Cytoflavin".

Acute toxicity of “Cytoflavin” was determined on linear white CBA mice in groups of 10 animals.

The drug was injected into the tail vein in increasing doses according to Wilcoxon-Litchfield and the LD ₅₀ index was determined - the dose of the drug that causes the death of 50% of the animals.

The results of the study are presented in table 9.

The results of the experiment showed that the preparation The word “Cytoflavin” filed an application No. “Cytoflavin” refers to the low-toxicity 98708522 for registration of a trademark in unions: LD ₅₀ is 1146 + 124 mg / kg. Russian Federation, filing date 05/19/98.

Table 1. The effect of the solution "Cytoflavin" and the prototype on aconitine and propranol arrhythmias in mice

The drug used in the experiment The number of mice with extrasystole,% Latent period to extrasystole, min The duration of extrasystole, min Number extrasystoles Aconitine 100.0 32 ± 2.8 * 19.6 ± 1.3 * 306 ± 14 * Aconitine ± "Cytoflavin" 25.0 21.3 ± 1.7 * 11.4 ± 1.5 * 127 ± 11 * Aconitine + Prototype 75.0 29.6 ± 2.2 * 19.2 ± 1.1 * 312 ± 15 * Propranol 100.0 18.6 ± 1.4 * 10.4 ± 1.1 * 359 ± 23 * Propranol + "Cytoflavin" 40,0 13.2 ± 1.2 * 6.2 ± 1.0 * 112 ± 9 * Propranol + Prototype 50,0 16.4 ± 1.2 * 10.5 ± 1.0 * 208 ± 20 *

Note: * - significance of differences at P <0.05

Table 2. Anti-ischemic effect of “Cytoflavin”

Solution* preparations The number of ventricular extrasystoles Tachycardia Ventricular fibrillation Frequency% Duration sec * Frequency occurrence of violations,% Duration sec * "Cytoflavin" 137 ± 12 * one hundred 42 ± 3 * 40 28 ± 5 Prototype 221 ± 15 * one hundred 90 ± 7 * 60 27 ± 3 The control 359 ± 24 * one hundred 127 ± 23 * 86 28 ± 4

Note: * - significance of differences at P <0.05

Table 3. The effect of “Cytoflavin” on the survival of animals in conditions of early postischemic arrhythmias

A drug number operated rats Survival after 60 minutes of myocardial ischemia number % "Cytoflavin" 12 12 one hundred Prototype 12 6 fifty The control 12 3 25

Table 4. The effect of “Cytoflavin” on mean arterial pressure (SBP) and heart rate (HR) in early conditions of early postischemic arrhythmias

Index A drug IN] myocardial ischemia, min 0 5 fifteen thirty 45 60 CAD% of the original "Cytoflavin" one hundred 57 ± 12 67 ± 2 * 72 ± 4 * 73 ± 1 * 73 ± 4 * Prototype one hundred 45 ± 8 59 ± 5 * 65 ± 7 * 67 ± 6 * 73 ± 7 * The control one hundred 31 ± 5 44 ± 5 * 53 ± 9 68 ± 12 73 ± 6 * Heart rate in% of the original "Cytoflavin" one hundred 94 ± 12 93 ± 6 * 95 ± 8 * 95 ± 8 * 96 ± 8 * Prototype one hundred 92 ± 16 90 ± 6 * 86 ± 5 * 84 ± 6 * 82 ± 5 * The control one hundred 85 ± 7 83 ± 4 * 82 ± 5 * 80 ± 6 * 75 ± 8 *

Note: * - significance of differences at P <0.05

Table 5 Hepatoprotective effect of "Cytoflavin" in comparison with the prototype

Indicators Experimental groups BUT (saline) B (cytoflvin) IN (prototype) G (without treatment) D (cytoflavin) E (prototype) Total protein, g / l 64 ± 2 65 ± 5 64 ± 6 40 ± 4 * 58 ± 4 * 46 ± 5 * Total lipids, g / l 3.7 ± 0.3 3.7 ± 0.3 3.5 ± 0.1 3.0 ± 0.3 * 3.4 ± 0.1 * 3.4 ± 0.1 * AST, mkat / l 0.60 ± 0.05 0.60 ± 0.06 0.60 ± 0.06 0.99 ± 0.12 * 0.73 ± 0.06 * 0.88 ± 0.12 * Alkaline phosphatase, mkat / l 0.69 ± 0.11 0.69 ± 0.12 0.69 ± 011 2.65 ± 0.24 * 0.73 ± 0.04 * 0.96 ± 0.14 * CF, mkat / l 0.74 ± 0.12 0.75 ± 0.11 0.73 ± 0.12 1.99 ± 0.20 * 0.72 ± 0.04 * 0.96 ± 0.11 * Thymol test, units 1.46 ± 0.04 1.48 ± 0.03 1.43 ± 0.12 5.77 ± 0.26 * 1.56 ± 0.12 * 2.88 ± 0.13 * ALT mkkat / l 0.20 ± 0.03 0.20 ± 0.02 0.25 ± 0.04 1.68 ± 0.11 * 0.32 ± 0.06 * 0.52 ± 0.08 * Liver glutathione, mg% 160 ± 5 155 ± 5 170 ± 12 70 ± 10 * 130 ± 10 * 100 ± 10 * Liver glycogen, mg% 2500 ± 100 2400 ± 90 2460 ± 110 850 ± 80 * 2000 ± 150 * 1800 ± 200 Hexenal sleep, min 25.0 ± 1.5 26.0 ± 1.8 24.0 ± 1.6 46.5 ± 2.5 * 27.0 ± 1.5 * 35.2 ± 1.0 * Total bilirubin, mmol / l 3.0 ± 0.3 3.0 ± 0.1 3.2 ± 0.2 6.8 ± 0.4 * 3.2 ± 0.2 * 4.2 ± 0.1 intact animals experienced animals

Note: * - significance of differences at P <0.05

AST - aspartate aminotransferase alkaline phosphatase - alkaline phosphatase KF - creatine phosphate ALT - alaminaminotransferase

Table 6. Renoprotective effect of "Cytoflavin" in comparison with the prototype, M ± m

Indicators Experimental groups BUT (saline) B (cytoflavin) IN (prototype) G (for treatment) D (cytoflavin) E (prototype) The body weight of rats, g 184 ± 6 183 ± 7 188 ± 11 154 ± 11 * 181 ± 11 160 ± 11 The relative mass of the kidneys, g 7.9 ± 0.9 7.8 ± 1.2 7.7 ± 1.0 11.2 ± 1.5 * 8.4 ± 0.7 10.8 ± 1.1 Daily diuresis, ml 3.7 ± 0.2 3.5 ± 0.3 3.5 ± 0.4 1.9 ± 0.3 * 2.8 ± 0.4 * 1.9 ± 0.5 Urine protein, mg / ml 4.4 ± 0.5 4,5 ± 0,4 4,5 ± 0,4 12.4 ± 1.3 * 6.0 ± 0.6 * 8.7 ± 0.8 * Urine Chloride, mg / ml 4.0 ± 0.3 3.8 ± 0.4 4.0 ± 0.3 1.2 ± 0.3 * 3.3 ± 0.3 * 2.5 ± 0.4 * Urine Sugar, mmol / L 1.5 ± 0.5 1.4 ± 0.3 1.6 ± 0.1 4.2 ± 0.3 * 2.4 ± 0.4 3.0 ± 0.5 Urea, mol / L 4.12 ± 0.34 4.22 ± 0.28 4.18 ± 0.32 12.35 ± 0.77 * 5.17 ± 0.11 * 8.77 ± 0.52 * Urine density 1.02 ± 0.006 1.02 ± 0.005 1.02 ± 0.004 0.91 ± 0.006 * 1.02 ± 0.003 * 1.00 ± 0.003 * Creatinine mmol / L 0.075 ± 0.012 0.0075 ± 0.012 0.073 ± 0.011 0.065 ± 0.08 * 0.047 ± 0.04 * 0.35 ± 0.09 * Lactate dehydrogenase, mmol / h / l 4.90 ± 0.26 4.77 ± 0.33 4.87 ± 0.31 8.08 ± 0.76 * 5.85 ± 0.55 * 7.93 ± 0.62 * intact animals experienced animals

Note: * - significant differences from intact animals at P <0.05

Table 7. The effect of the content of лю-methylglucamine on the pH and stability of "Cytoflavin"

Option The content of Ν-methylglucamine,% Characterization of the stability of the dosage form Note BUT 0 Precipitation occurs during storage - acid hydrolysis occurs Not suitable for use B 5,0 Precipitation occurs during storage - acid hydrolysis occurs Not suitable for use IN 10.0 The solution is stable during storage, but has an acidic pH. Not suitable for intravenous administration G 15.7 The solution is stable during storage. Suitable for use d 17.3 The solution is stable during storage. Suitable for use E 20,0 The solution is stable during storage, but has an alkaline pH value. Not suitable for intravenous administration F 25.0 The solution during storage turns black alkaline hydrolysis occurs Not suitable for use

Table 8. The effect of sodium hydroxide on the pH of the solution "Cytoflavin"

Option The amount of sodium hydroxide,% PH value of the solution Characterization of the resulting solution Note BUT 0 4.01 Incomplete dissolution of succinic acid The dosage form is not suitable for use B one 4.88 Incomplete dissolution of succinic acid The dosage form is not suitable for use IN 2 5.7 Acidic pH The solution is not suitable for intravenous administration. G 3.3 6.4 Physiologically acceptable value The solution is suitable for intravenous administration. D 3,7 7.2 Physiologically acceptable value The solution is suitable for intravenous administration. E 5 8.3 Alkaline pH The solution is not suitable for intravenous administration.

Table 9. Acute toxicity of Cytoflavin

The dose of "Cytoflavin", mg / kg 794 1000 1260 1580 2000 Mice survived 10 6 4 0 0 The mice fell 0 4 6 10 10

CLAIM

Claims (1)

An injection drug with a cytoprotective effect, containing an active composition of natural metabolites and coenzymes in a solvent, characterized in that it contains succinic acid and riboxin (inosine) as natural metabolites, and nicotinamide and riboflavin-5-mononucleotide sodium as coenzymes, as solvent water for injection, and also additionally contains sodium hydroxide and Ν-methylglucamine in the following ratio, wt.%: succinic acid 9.5 -10.5 Riboxin (Inosine) 1.9-2.1 Nicotinamide 0.95-1.05 Riboflavin-5- sodium mononucleotide 0.19-0.21 Sodium hydroxide 3.3-3.7 Ν-Methylglucamine 15.7-17.3 Water for injections Up to 100 Eurasian Patent Organization, EAPO