RU2205637C1 - "corsilin" as hepatoprotective product - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: hepatoprotective product is suggested in tabletted form containing a substance covered with a multi-layer membrane. The core of tablets contains additional substances except silimarine as an active product at the quantity of 10-20% against core's weight. These tablets are of high velocity to release an active substance. EFFECT: higher efficiency of hepatoprotective action. 1 dwg, 3 tbl

Description

 The invention relates to medicine and relates to the production of a hepatoprotective agent, the active principle of which is silymarin, a flavonoid compound isolated from the fruits of milk thistle.

 Flavonoid compounds have a phenylchromanone structure. Their hepatoprotective effect is explained by antioxidant activity, stimulation of protein synthesis, and normalization of phospholipid metabolism.

 Known tool containing salts of polyhydroxyphenylchromanones (US p. 4061765, A 61 K 31/335, 1977). It is presented in various dosage forms, including in the form of coated tablets.

 Closest to the proposed drug in terms of technical nature and the achieved result are products containing as the active substance the sum of flavonoids from the fruits of milk thistle. These funds are made in the form of dragees, tablets "Silibor", "Legal", "Karsil" (MD Mashkovsky. Medicines, M., 1993, v. 1, p. 611-612).

 The aim of the present invention is to obtain a domestic dosage form of silymarin in the form of coated tablets with bioavailability, improved by increasing the rate of release of the active substance.

 This goal is achieved using the proposed hepatoprotective agent called "Karsilin". The tool is made in the form of tablets containing a core coated with a multilayer shell. The tablet core contains the active substance silymarin in an amount of 10-20% by weight of the core, as well as excipients: microcrystalline cellulose, lactose, corn starch, crospovidone, polyvinylpyrrolidone, sodium bicarbonate, talc and stearic acid and / or its salts.

To obtain the nuclei of Karsilin tablets, the following components are used,% of the mass of the nucleus:
Silymarin - 10-20
Microcrystalline cellulose - 15-30
Corn starch - 15-30
Crospovidone - 1-5
Polyvinylpyrrolidone - 5-15
Sodium bicarbonate - 0.1-1
Talc - No more than 3
Stearic acid and / or its salts - Not more than 1
Lactose - Rest
To obtain Karsilin tablet cores, the active substance, microcrystalline cellulose, lactose, part of starch, crospovidone are mixed and granulation is carried out in a fluidized bed installation. As the granulating liquid, a solution of sodium bicarbonate in an aqueous solution of polyvinylpyrrolidone is used. The granulate obtained is dried, dry granulated and dusted with the remaining crospovidone and starch, talc, stearic acid and / or its salts. The resulting mixture was tabletted. The tablet cores have an average weight of 0.25 g.

The core of the tablets is coated with a shell having the following composition,% of the total shell weight:
I layer
Etazel - 0.05-0.4
Polyethylene glycol 6000 - 0.05-0.5
II layer
Sugar - 70.0-90.0
Polyvinylpyrrolidone - 0.2-0.6
Sodium carboxymethyl cellulose - 0.3-1.2
Twin 80 - 0.08-0.3
Talc - 5.0-9.0
Titanium dioxide - 0.3-3.0
Aerosil - 0.15-0.5
III layer
Sugar - 4.0-9.0
Opalux AS 9840 Brown - 1.0-3.0
IV layer
Polyethylene glycol 6000 - 0.06-0.2
Glycerin - 0.005-0.03
The coating process is carried out in a coating pan. Get tablets "Karsilin" with a brown shell and with an average weight of 0.45 g

 A study of the experimental preparation - tablets "Karsilin" in comparison with the dragee "Karsil" company "Sofarma", Bulgaria.

 A study of the bioavailability of the compared preparations was carried out on white Wistar male rats weighing 190-210 g. Rats were given a suspension of the ground mass of the compared tablets and dragees at a dose of 600 mg / kg in terms of the active substance. A suspension was prepared immediately before administration to the animals. Blood samples were taken after decapitation of rats under ether anesthesia at 1, 2, 3, 4, 5, 6, and 8 hours after drug administration. For each time point, 8 animals were used.

Appropriately prepared blood samples were analyzed on a Milichrom-5 liquid chromatograph with a column filled with Diasorb C ₁₆ . As the mobile phase, the solvent system acetonitrile-water-glacial acetic acid (49: 50: 1) was used. Detection was carried out at a wavelength of 280 nm. The calculation of the content of the drug in blood serum was carried out according to the calibration schedule. Since silymarin is the sum of various flavonolignans that have different solubilities in water, the rate and degree of absorption and elimination, when conducting this study, we took into account only one of the peaks in the chromatograms of the solution of the working standard of silymarin (maximum), the results of which differed more reproducibility.

 The results of a quantitative assessment of the content of silymarin in blood serum by oral administration of the compared dosage forms are presented in the drawing. Table 1 shows the main pharmacokinetic parameters calculated taking into account the two-chamber model, which describes the obtained concentration curves.

скорость всасывания, чем применение драже "Карсил".To assess the pharmacokinetics and bioavailability, we calculated the constants of the rates of absorption, distribution, elimination, the maximum concentration of the drug in the blood and the time it was reached, half-suction and half-life, and average retention time. From the results it follows that the concentration profiles of silymarin for the two dosage forms differ slightly. The maximum amount of the drug in the blood when using both dosage forms is observed after 3 hours. The use of Karsilin tablets provides a slightly higher level of concentrations of active substances in the blood after 3 and 4 hours compared with Karsil. However, the concentration levels do not significantly differ (P> 0.05) and after 3 hours (19.1 ± 3.8) and (16.5 ± 2.9) μg / ml, after 4 hours - (16.2 ± 2.3) and (11.3 ± 1.7) μg / ml, respectively. The constants of the suction rate and half-suction time are K ₀₁ = 0.735 h ^-1 , T _{0.5; 01} = 0.94 h and K ₀₁ = 0.557 h ^-1 , T _{0.5; 0.1} = 1.2 h, respectively. The results allow us to conclude that the use of Karsilin tablets provides

suction rate than the use of tablets "Carsil".

 After reaching the maximum concentration, silymarin is also distributed in organs and tissues at approximately the same rate. The average retention time for the compared dosage forms is also almost the same - 5.8 and 5.5 hours. Calculation of the constants of the transition of the drug into the peripheral chamber from the central and vice versa shows that silymarin does not accumulate in the body.

 At the same time, the areas under the pharmacokinetic curves are different. The relative bioavailability of silymarin when using the experimental dosage form is 126% compared with the comparison drug. The increase in bioavailability is due to the fact that the use of Karsilin tablets ensures a more rapid entry of the drug into the systemic circulation and, as a result of this, a greater amount of the drug enters the bloodstream.

 To confirm the obtained data, we studied the pharmaceutical availability of silymarin from the studied dosage forms by the "rotating basket" method in accordance with the requirements of Global Fund XI. The data obtained are presented in table 2. From the table it is seen that the rate of release of silymarin from the experimental dosage form is 30% higher than the rate of release of silymarin from the reference drug (after 3 hours of observation). This is consistent with the results of a pharmacokinetic experiment.

 Thus, a comparative study of the pharmacokinetics and bioavailability of two dosage forms containing the sum of flavolignans of Milk Thistle, Karsilin tablets and Karsil tablets, showed that Karsilin tablets slightly exceed the comparison drug in terms of bioavailability. At the same time, with the use of Karsilin tablets, the drug is more rapidly absorbed into the blood, which provides higher levels of its concentration in the blood and somewhat greater bioavailability. The increase in relative bioavailability is due to the fact that Karsilin tablets provide a high rate of release of active substances. However, due to the fact that the differences in bioavailability are insignificant, these dosage forms can be considered bioequivalent.

 The study of the specific hepatoprotective activity of Karsilin tablets and Karsil tablets was carried out on 84 randomly bred adult white rats of both sexes weighing 143.3-215.0 g. All animals were divided into 7 groups, each with 6 females and 6 males. The preparations were administered orally to animals using a probe in the form of a suspension at a dose of 200 mg / kg.

The specific hepatoprotective activity of Karsilin tablets and Karsil tablets was studied by means of therapeutic and prophylactic administration of drugs to rats using the acute hepatitis model. As a hepatotropic poison used a 50% solution of carbon tetrachloride (CCl ₄ ) in liquid paraffin in a dose of 3 ml / kg of animal body weight. The drugs were administered within 6 days before the hepatotoxin was administered, and then, in parallel with the studied drugs, carbon tetrachloride was orally administered three times a day one hour after the drug was administered.

 A model of drug hepatitis was created by administering paracetamol at a dose of 700 mg / kg every other day for 2 days.

 Carbon tetrachloride is an industrial hepatotropic poison. Paracetamol is widely used as an antipyretic, analgesic and anti-inflammatory agent, including in children's practice. Its side effect is hepatotoxicity.

 The control groups of rats were injected with carbon tetrachloride and paracetamol in the same doses. Intact animals received 1 ml of water.

 Animals were killed one day after the last dose of the toxicant. At autopsy, the liver was weighed to determine the relative weight. It is known that in acute hepatitis, the size of the liver can increase dramatically. Therefore, it was of interest to partially evaluate the hepatoprotective effect of Karsilin in comparison with Karsil by changing this indicator.

In order to assess the effect on the functional activity of the liver of the compared preparations, the following biochemical studies were carried out:
- total protein was determined in a unified manner with a biuret reagent;
- albumin was determined by reaction with bromocresol green;
- urea was determined by the color reaction with diacetylmonooxime;
- thymol test was performed by the method of Huergo and Popper;
- cholesterol was determined by the Ilk method;
- bilirubin fractions were determined by the diazomethod according to Yendrashik, Cleggorn, Grof;
- the activity of the most important aminotransferases - alanine aminotransferase (A _l AT) and aspartate aminotransferase (A _with AT) was determined by the Wrightman-Frenkel method;
- the activity of alkaline phosphatase (ALP) in blood serum was determined by the reaction of hydrolysis with p-nitrophenyl phosphate.

The relative liver mass in animals with CCl ₄ hepatitis was 27.59% in males and 30.25% more in females than in intact rats.

 Paracetamol hepatitis gave an increase in liver in relation to body weight by 12.53% in females and by 10.88% in males in comparison with healthy rats.

The use of Karsil in animals with the CCl ₄ hepatitis model led to a decrease in the relative mass of the liver in comparison with animals of the control group by 4.69% in females and by 3.91% in males. Against the background of Karsilin tablets, under the same conditions, the relative mass of the liver decreased by 13.03% in female rats and by 12.27% in male rats. That is, “Karsilin” in toxic hepatitis reduced the size of the liver by 8.3% more than “Karsil”.

 Against the background of paracetamol hepatitis in relation to intact animals, “Karsilin” and “Karsil” increased the relative mass of the liver by an average of 10%, and in comparison with control rats - by an average of 2%.

 Thus, the measurement of the relative mass of the liver showed that carbon tetrachloride compared with paracetamol gives more pronounced hepatitis, against which the effect of Karsilin tablets is 8% higher than that of Karsil tablets.

It was found that carbon tetrachloride reduced the content of total protein in blood serum by 36% in females and 39.9% in males. Karsilin and Karsil increased the protein content in the blood of female rats by 12.68 and 15.81%, respectively, in comparison with the protein content in the blood of control female rats (with the CCl ₄ hepatitis model). In male rats, under similar conditions, the blood protein increased by 17.54% with Karsilin and 17.79% with Karsil.

The level of albumin in the blood of females treated with Karsil for therapeutic and prophylactic purposes was 30.59%, and those treated with Karsilin were 24% higher than in the control groups of rats (CCl ₄ hepatitis). In males, this indicator “Karsilin” increased by 29.73%, “Karsil” - by 20.7% compared with the group of untreated male rats.

The content of globulins in the blood of females increased by 3.3% when using both Karsilina and Karsila. In males, the level of globulins increased against the background of Karsil by 14.9%, against the background of Karsilin by 7.14%, that is, in males the globulin fraction of proteins was slightly higher than in females in comparison with the control group of animals (CCl ₄ hepatitis). The albumin / globulin coefficient was similarly changed.

 Thus, "Karsilin" and "Karsil" gave a close, but not very pronounced effect on the restoration of total protein and its fractions in the blood of both females and males.

 Against the background of paracetamol hepatitis in male rats, the albumin fraction against the background of Karsil increased by 17.6%, against the background of Karsilin by 28.4%, the level of globulins increased by 2.7-5.3%, in females albumin and globulin changed in the same way.

 Therefore, against the background of paracetamol hepatitis, the restoration of the level of protein in the blood with the use of Karsilin is on average 10% higher than with Karsil.

With paracetamol hepatitis, the therapeutic and prophylactic effect on the protein-synthetic function of the liver in both “Karsilin” and “Karsil” was higher than with CCl ₄ hepatitis.

The thymol test for CCl ₄ hepatitis increased in females by 560%, in males by 448%, and against paracetamol hepatitis - by 437.5% and 342%, respectively, in females and males compared to intact rats.

Karsil reduced the sample against paracetamol hepatitis to 115%, and Karsilin to 128.8%, that is, both drugs worked almost the same. Against the background of CCl ₄ hepatitis, Karsil reduced the thymol test by only 24%, and Karsilin by 22% in comparison with the control, that is, almost equally, but in comparison with healthy rats this indicator was quite high as in females (402.5 and 413.75%), and males (318 and 300%). In general, the thymol test index both drugs reduced by an average of 20-25%.

The urea content in toxic hepatitis decreased by 41.7% in females and by 46.1% in males. Against the background of paracetamol, detoxification function of the liver decreased to a lesser extent. Karsil and Karsilin increased the urea content in the blood serum of rats with the CCl ₄ hepatitis model by 38–40% in females and by 43.7–42.3% in males. Against the background of paracetamol hepatitis, the blood urea content increased in comparison with the control by 23.3 and 22.5% in females and by 31.2 and 29.0% in males with the use of Karsil and Karsilin, respectively.

Both drugs slightly increased cholesterol against CCl ₄ hepatitis (1-3%), slightly more against paracetamol hepatitis (13-14%), that is, with drug hepatosis, both drugs normalized the rate of fat metabolism to a greater extent.

The content of total bilirubin in acute hepatitis increased by 105.9-154.9% in females and males. The administration of Karsil and Karsilin reduced the level of bilirubin in animals of both sexes with both hepatitis models by an average of 50%. The concentration of direct bilirubin in CCl ₄ hepatitis increased by 400.0 and 278.3% in females and males, respectively. “Karsil” and “Karsilin” reduced it by 2 times in both females and males. Against the background of paracetamol hepatitis, the effect of both drugs manifested similarly.

 Thus, the concentration of total and direct bilirubin in the blood serum of rats treated with Karsil and Karsilin decreased almost equally, which indicates the restoration of pigment metabolism under the influence of drugs.

The activity of transaminases on the background of CCl ₄ hepatitis increased. To AT _And this increase was 200.0% for females and males 184.0% for AT A _l - y 452.6% 430.0% females and males respectively. When using both drugs against the background of CCl ₄ hepatitis, the activity of A _with AT decreased slightly, and against the background of drug hepatitis - by 3.5-4 times in females and 5-17 times in males. "Karsilin" reduced the level of A _with AT to a greater extent. The level of A _l AT decreased by 1/3 with the introduction of "Karsil" and "Karsilina" in rats of both sexes with the model CCl ₄ hepatitis. Against the background of paracetamol hepatitis, the level of A _l AT decreased by 3-4 times in females and males with the use of both drugs. The de Ritis coefficient (ratio of A _to AT / A _l AT) in females of all experimental groups was below normal (1.33). Amino acid balance increased significantly with the introduction of "Karsil" and "Karsilin" to rats with paracetamol hepatitis, but not completely.

 The therapeutic and prophylactic use of drugs reduced the activity of alkaline phosphatase by an average of 2 times in both females and males.

The results of biochemical studies of blood serum showed that the effect of Karsilin tablets and Karsil tablets on the metabolic, metabolic, synthetic, detoxification, choleretic functions of the liver is almost equivalent, although it is less pronounced against CCl ₄ hepatitis than against paracetamol hepatitis A. Differences in the effect of drugs on many indicators are unreliable and indicate a slight decrease in the processes of cytolysis and cholestasis in the liver.

 At autopsy, the state of the liver was visually assessed. In intact rats, the liver was dark red in color, elastic, with a smooth shiny surface, the capsule of the liver was transparent. The tissue of the liver on the cut was homogeneous, the vessels were filled with blood.

In rats with a model of acute CCl ₄ hepatitis, the liver was light brown, dull, friable. Liver tissue was easily injured.

 In animals with a model of acute paracetamol hepatitis, the liver tissue was rather dense, dark red in color, brown blotches in the form of dots of different sizes were visible on the section.

In rats administered Karsilin and Karsil against the background of acute CCl ₄ hepatitis, the liver tissue was denser than in rats with a clean model of CCl ₄ hepatitis. The color of the liver was reddish brown.

 The liver of animals that received drugs on the background of acute paracetamol hepatitis, outwardly looked like healthy, but in some places brown specks were seen on the surface, indicating partial toxic effects of large doses (700 mg / kg) of paracetamol.

 Conducted microscopic studies using "Karsilin" and "Karsil" at a dose of 200 mg / kg against the background of the subsequent administration of paracetamol at a dose of 700 mg / kg. It has been established that signs of subacute drug-induced focal hepatitis with predominantly portal localization persist in the liver. At the same time, the appearance of microstructural equivalents of the development of compensatory-adaptive processes in the liver is noteworthy. They consist of an increase in cell size, activation of their plastic function, nuclear hyperchromia, chromatin contourability, nucleolus swelling, the appearance of binuclear hepatocytes, and activation of cell cooperatives.

 Microscopic studies were carried out using Karsilin and Karsil at a dose of 200 mg / kg, followed by the introduction of carbon tetrachloride in the form of a 50% solution in liquid paraffin at a dose of 3.0 ml / kg. It was found that with the use of Karsil followed by the introduction of carbon tetrachloride in the liver of rats, a diffuse subacute toxic alternatively-exudative parenchymal-interstitial hepatitis with partial nodular transformation and hyperplasia of certain groups of hepatocytes was diagnosed. In this series of experiments, the hepatoprotective effect of Karsil was not found, but with a certain degree of conditionality, the presence of nodular transformation and focal hyperplasia of liver cells can be considered a certain tendency to this.

 In all series of experiments, no fundamental differences were found in the pathology of the liver and the development of reparative processes (they proceeded according to the type of substitution) in rats of both sexes both with the use of Karsilin and against the background of Karsil.

 In addition, a comparative study of the "acute" and chronic toxicity of Karsilin tablets and Karsil tablets was conducted.

 Studies of the general toxic effect of the preparations were carried out on 72 nonlinear white mice of both sexes weighing 25-30 g and on 80 outbred white rats of both sexes weighing 150-220 g.

 The “acute” toxicity of the preparations was determined by the Kerber method.

 The suspension of preparations was administered orally to mice using a needle with olive at doses of 4000, 4500, 4600, 4700, 4800, 5000 mg / kg. Drugs were administered during the day every 3-4 hours. Animals were divided into 6 groups of 6 mice each. Part of the mice received a suspension of the experimental drug, another part - a suspension of the comparison drug. During the first day, the mice were closely monitored, noting the behavior, consumption of food, water, as well as the general condition. Dead mice were opened and examined macroscopically. Surviving animals were observed for 2 weeks.

 When studying the chronic toxicity of drugs, all rats were divided into 4 groups of 20 rats each (10 females and 10 males). Two groups of animals received daily for 2 months an hour before feeding an aqueous suspension of Karsilin tablets: the first group at a dose of 20 mg / kg, the second group - 200 mg / kg. The third group was administered under the same conditions a suspension of dragee Karsil at a dose of 200 mg / kg. The fourth group was poured into the stomach through a tube of 1 ml of water. The amount of food and water for rats was not limited. The condition of the animals was visually assessed daily.

 The general toxic effect of Karsilin tablets was evaluated by their effect on various indicators and body functions. During the first month, the animals were weighed weekly before feeding. The last weight control was carried out before slaughter. In the course of the experiment, we monitored the appearance of the animals, the work of the heart, respiratory system, the state of the hairline, and motor activity. The effect of Karsilin tablets on the central nervous system was evaluated by the open field method. The resulting animal behavior indicators made it possible to judge the effect of Karsilin on physical activity, research and emotional activity, reflecting the central nervous system. Each animal was tested once.

 The nature and degree of toxic effect of Karsilin tablets in comparison with Karsil tablets during long-term use was evaluated by hematological, biochemical and pathological studies.

 Pathomorphological studies were carried out macroscopically at the face, for microscopic studies, organs were prepared accordingly.

With the introduction of the first portions of toxic doses of Karsilin and Karsil, the behavior of white mice was somewhat inhibited for 2-5 minutes, in the next 3-4 hours their motor activity, behavior, appearance did not differ from these indicators in mice of the control group. The heart rate before the administration of the drugs ranged from (450.0 ± 10.2 ^x ) to (500 ± 12.5 ^x ) beats / min (x - changes in the series are significant, P≤0.05). 5-10 minutes after the intragastric administration of suspensions of “Karsilin” and “Karsil” in a volume of 0.5 ml, the rhythm increased from (560.0 ± 8.5 ^x ) to (640.0 ± 16.6 ^x ) beats / min, and after 40-60 minutes the rhythm was restored. The number of initial respiratory movements was on average 146.7 ± 14.8 ^x before administration of the drugs, 160.0 ± 5.4 ^x after 5-10 minutes after administration, then the respiratory rhythm was restored. The mice 'intake of food and water decreased. The inclination of the hairline did not change, convulsions were not observed, the coordination of movements was not disturbed.

With the introduction of drug suspensions in a volume of 0.5 ml 4-6 times at intervals of 3-4 hours (maximum dose), the behavior of the animals changed. Their motor activity decreased, the rhythm of heart contractions and the number of respiratory movements gradually decreased until they completely stopped. Surviving mice were monitored for 2 weeks. One day after the administration of toxic doses, their appearance and behavior did not differ from the appearance and behavior of control mice. The dead mice were opened. Hyperemia was visible in the internal organs, the intestines contained the drug and was significantly expanded due to the large volume of injected studied drugs. Of the mice that received toxic doses of Karsilin, 16 died, and 11 of them died on the first day after the last dose of the toxic dose was administered - mainly after 5-8 hours. The remaining 5 mice died on the second day. When calculating LD ₅₀ for Karsilin tablets, a result of 4666.67 mg / kg was obtained.

With the introduction of toxic doses of the Karsil dragee out of 36 mice, 12 died. The picture of their death was similar to the picture of the death of mice from the maximum doses of Karsilin. The calculation showed that for Karsil, the LD ₅₀ value is 4775.0 mg / kg, that is, the toxicity of Karsil is slightly less than that of Karsilin. The toxic effect of tablets and dragees on females and males was the same, that is, sexual sensitivity to drugs was absent.

 Thus, a study of the “acute” toxicity of Karsilin tablets compared to the Karsil tablets showed that, according to the classification of danger, both drugs can be classified as moderately hazardous substances (class III), and according to the classification of toxicity, they can be classified as relatively harmless ( VI class of toxicity).

 When studying the chronic toxicity of the drugs within 2 months of animal soldering, not a single rat died in either the control or experimental groups. The first 7-10 days of the rat during the administration of the suspension "Karsilina" and "Karsila" showed some anxiety, alertness, responded to pain by hyperreaction. In the future, during the course of the study, these phenomena decreased, the animals behaved calmly both at the time of working with them and throughout the day. Aggression was not observed either among females or among males. Appearance of experimental rats was normal, there was no change in the color of the hair, the inclination of the hairline. The color of the noses and paws was the same in comparison with the animals of the control groups. Animals consumed food in unlimited quantities, shipments were in slightly larger quantities than in control rats, and in general almost always decorated. The animals consumed water in an amount of 200 to 300 ml per day per group consisting of 10 rats.

 In the first week of the experiment, the animals practically did not gain weight, by the end of the first month the weight gain was 11-17%, during the second month of using the drugs, the animal weight stabilized, while in the control groups it increased by 7-10%.

 The results of monitoring the effect of drugs on the central nervous system are shown in table 3.

 The motor activity of males with the introduction of "Karsilin" and "Karsil" decreased in contrast to the control groups by an average of 15%, and in females the number of sectors passed increased by 12-30%. In both groups of animals, approximately cognitive activity decreased. The number of racks against the background of the introduction of both drugs decreased by 60-75%.

The time of washing (grooming) in the control groups was males and females, respectively (16.33 ± 2.76 ^x ) and (16.17 ± 2.9 ^x ) s. When using “Karsila” this indicator was (8.83 ± 0.83 ^x ) s (males) and (9.50 ± 0.5 ^x ) s (females). With the introduction of Karsilin in the same dose, males washed themselves (8.83 ± 2.36) s, and females (16.5 ± 1.65) s. The hardening time against the background of both drugs in males increased to a greater extent (by 43-100%), and in females to a lesser extent (by 4-24%).

 In this regard, we can say that in the doses used, both drugs reduced emotional activity. A general assessment of rat testing in an “open field” suggests that Karsilin, like Karsil, acts on the central nervous system of rats in a depressing (calming) manner.

The results of clinical blood tests showed that the content of red blood cells in males and females treated with Karsilin and Karsil practically did not differ in the experimental or control groups of animals. Hemoglobin (Hb) was 20 g / L higher in animals treated with Karsilin at a dose of 20 g / L than in rats of other groups, both experimental and control. The erythrocyte sedimentation rate (ESR) in all animals did not deviate from the norm, but in males, Karsilin increased ESR by 2.2 times in comparison with Karsil, although these changes were unreliable. The percentage of reticulocytes in the blood, as well as the hematocrit, were within acceptable limits when using both drugs, but their number was within the acceptable limits for both males and females. The osmotic resistance of red blood cells was 0.35-0.45 units, and its increase against the background of "Karsilin" in male rats (in both doses) to 0.45 units. was reliable in comparison with the control and "Carsil" and did not go beyond the norm. In females, the osmotic resistance of red blood cells did not change significantly. The increase in platelet count against the background of “Karsil” and “Karsilin” was insignificant (by 2-3%) and did not go beyond the norm (in animals (522.0 ± 33.0) 10 ⁹ / l). The white blood cell count was (4.10 ± 0.19 ^x ) 10 ⁹ / L - (4.46 ± 0.19 ^x ) 10 ⁹ / L in males and (4.02 ± 0.08 ^x ) 10 ⁹ / L - (4.52 ± 0.47 ^x ) 10 ⁹ / l in females, including against the background of “Karsilin” at a dose of 200 mg / kg, the number of leukocytes was (0.18-0.22) 10 ⁹ / l more, than when using "Karsil". Compared with the control, changes in the number of leukocytes were unreliable. In the leukocyte formula, the changes are unreliable, with the exception of the percentage change in the content of eosinophils with the introduction of Karsilin. The content of eosinophils in the blood of males and females against the background of Karsilin increased significantly in relation to Karsil, but did not go beyond the norm. The number of segmented neurophiles ranged in males from (20.69 ± 0.46 ^x )% (against the background of Karsilin at a dose of 20 mg / kg) to (22.06 ± 0.75 ^x )% (against the background of Karsil) . The percentage of lymphocytes was from 65.56 ± 2.17 ^x (in males on the background of Karsil) to 70.54 ± 1.42 ^x (on females on the background of Karsil). The use of both drugs did not give rough shifts in the leukogram in comparison with the control group of rats. Moreover, changes in the clinical blood parameters of animals with the use of "Karsilin" and "Karsil" did not differ significantly and, in addition to the number of eosinophils, was not significant. All clinical blood counts with prolonged use of "Karsilin" and "Karsil" did not go beyond the norm.

During the study of urine composition, protein, glucose, ketones, hemoglobin, and bile pigments were not detected in either the control or experimental groups of animals. The specific gravity of urine averaged (1.006 ± 0.0005 ^x ) g / ml. The urine pH in the control group was 6.34 ± 0.16 ^x , against the background of "Karsil" (200 mg / kg) in females - 6.65 ± 0.21 ^x , in males - 7.45 ± 0.11. When using "Karsilin" at a dose of 20 mg / kg, the pH was 6.98 ± 0.08 ^x and 7.05 ± 0.14 ^x in females and males, respectively. With an increase in the dose of Karsilin to 200 mg / kg, the urine pH in females increased to 7.07 ± 0.19 ^x , and in males to 7.45 ± 0.15 ^x .

By the 3rd hour, urine collection under water load in rats treated with Karsilin tablets was (4.5 ± 0.05 ^x ) ml (females) and (4.3 ± 0.04 ^x ) ml (males).

Against the background of the “Karsil” dragee, the diuresis in comparison with the “Karsilin” was almost the same for both females and males. Urination in control rats was (3.6 ± 0.05 ^x ) and (3.8 ± 0.04 ^x ) ml in females and males, respectively.

 Thus, it was found that prolonged use of both studied drugs slightly increased diuresis, and almost equally.

 Microscopic analysis of urine sediment showed the absence of red blood cells, hyaline and granular cylinders, renal epithelium, and microflora in it. In the urine sediment of some rats in all groups (both control and experimental), a single urinary tract epithelium was determined - no more than 3-4 cells in the field of view. White blood cells in urine sediment were present in an amount of 1-4 in animals of all groups. The amount of mucus in the urine was slight or moderate. Of the salts found mainly oxalates and in some places urates.

 Thus, a clinical analysis of the composition of urine showed that the Karsilin tablets, like the Karsil tablets, do not cause renal pathology, can shift the pH of the urine to the alkaline side and have a small diuretic effect.

The introduction of "Karsilin" in comparison with "Karsil" to a small extent and unreliably lowered the level of glucose in both females and males, in contrast to the control group. Changes in the content of total protein and protein fractions tended to increase with the introduction of both "Karsil" and "Karsilin", and differences in their action were unreliable (P ₂ > 0.05). The thymol test in females treated with Karsil decreased by 46.25% compared with the control, and against the background of Karsilin in the same dose - by 23.75%. A 10-fold reduction in the dose of Karsilin resulted in a 10% decrease in the thymol test. In males, this indicator decreased to a lesser extent and almost unambiguously (on average by 20%). In a comparative assessment of both drugs on the thymol test value, the values were unreliable.

 Both drugs significantly (P≤0.05) increased the detoxification function of the liver, which is seen by an increase in the level of urea in the blood, but Karsilin did this to a greater extent and reliably with respect to Karsil. Serum cholesterol decreased with the use of Karsil in females by 13.25%, in males by 10.59%. "Karsilin" in both doses (20 and 200 mg / kg) reduced this indicator by 7% on average. With respect to control, the changes were significant.

 The content of total and direct bilirubin in the blood serum of rats decreased, and the percentage decrease in total bilirubin in both females and males was 4-5% lower with the administration of Karsilin than with the Karsil background. In general, indicators indicate a favorable effect of drugs on lipid metabolism.

With the introduction of Karsilin at a dose of 200 mg / kg, the activity of transaminases decreased in females by 4.40% for A _with AT and 6.72% for A _l AT, and in males - by 7.88 and 11.96%, respectively. Against the background of "Karsil" in the same dose, the activity of A _with AT was lower by 8.75%, A _{d of} AT - by 10.45%, and in males - by 4.25 and 8.22%, respectively. With the introduction of Karsilin at a dose of 20 mg / kg, the changes were less pronounced or absent. The activity of alkaline phosphatase in the blood serum of rats against the background of "Carsilin" decreased in females by 23-30%, in males by 15.66-23.14%. With the introduction of "Karsil" this indicator decreased by 36.77% (females) and 25.63% (males).

 When comparing the activity of “Karsilin” and “Karsil,” the changes are unreliable, that is, their activity against the choleretic and anticholeretic functions of the liver is close.

 Thus, biochemical studies of the blood serum of rats, received for two months in the form of a suspension of the Karsilin tablet in comparison with the Karsil tablet, showed that in the metabolic, detoxification, and metabolic functions of vital organs (liver, kidneys) against their background positive changes occurred (activation of functions), and the changes against the backdrop of Karsilin in relation to Karsil were unreliable in many respects, that is, very close.

 Macro and microscopic studies of the internal organs of animals were carried out after a long administration of Karsilin tablets.

 When slaughtering, animals were dissected, internal organs were examined. Some organs were weighed. Significant deviations in organ mass as a percentage of body weight were not detected.

 According to the location of the internal organs in animals of all groups, the norm is ascertained. Tissues of the chest, abdominal cavity are pale pink, shiny, smooth, no fusion sites. The heart is dark red with a little blood. There is no visible pathology in the cavities of the heart. The pericardium is easily separated and has normal humidity. Lung tissue with foamy contents. The stomach is filled with a small amount of food. When cut along a large curvature, a folded gray-pink gastric mucosa is visible. Patency of the gastrointestinal tract is not impaired. The intestine is partially filled with food masses, its tone is reduced in all rats equally.

 The liver has a red-brown color, its surface is smooth, covered with a serous shiny membrane. The liver tissue is dense, in the context of homogeneous, dark red in color with moderate blood vessels. The pancreas is grayish-red in color, lobed, granular, covered with a shiny capsule. Bile in experimental rats is more transparent and lighter than in animals of the control groups.

 The shape of the kidneys is correct, bean-shaped. The kidneys are dark red in color, with a dense consistency. Outside, a fibrous capsule, which is easily removed from the kidneys of all animals. In almost all rats, the kidneys are surrounded by fatty tissue in the form of a capsule. On sections of the kidneys, the cortical and cerebral layers are visible. The renal pelvis is free. The bladder in some animals is slightly full. When cut, the mucous membrane is folded, clean.

 In almost all rats, the spleen is dark cherry in color, the surface is granular, white and red pulp are determined in the section, and the vessels are dilated.

 We studied the micromorphology of the internal organs of rats after a 2-month administration of Karsilin and Karsil at a dose of 20 and 200 mg / kg of animal body weight.

 Liver. The central veins are moderately full-blooded. The beam structure of the hepatic lobules is preserved. Cells have clear boundaries, well-contouring nuclei with chromatin grains. Plethora is observed only in those sections of the sinusoidal capillaries that are adjacent to the central veins. Elongated endothelial cells. Disse spaces are not defined. The topology of Pit-cells, stellate reticuloendotheliocytes, in the structure of the hepatic lobule is not changed. In the cytoplasm of both light and dark hepatocytes, granularity is detected. Cells in a state of hydropic protein dystrophy, as well as in a state of fatty metamorphosis, are absent. Vessels of triads with moderate plethora in the lumens. In the stroma of the portal tracts - the usual cellular composition. Among hepatocytes, binuclear cells can be found.

 Stomach. The integument epithelium is saved. It evenly lines the gastric ridges, extending to the gastric fossa. Noteworthy is the slightly increased content of mucin in the cells of the integumentary epithelium and on its surface. The formula of the gland, that is, the ratio between the individual glandulocytes of the fundus glands, is not changed. The vessels of the own plate of the mucous membrane are moderately full-blooded.

 Small intestine. The villi are thin, lined mainly by limbic enterocytes. The number of goblet cells is not increased. The epithelium of villi, crypts is preserved in all observations. The cellular components of lymphoid follicles (Peyer's patches) are not changed either quantitatively or qualitatively. The vessels of the microvasculature of the mucous plate are moderately full-blooded.

 The kidneys. The glomerular capillaries are moderately full-blooded. The gaps of the capsules are free. Synechia between the visceral and parietal leaves of the capsule was not found. Proliferation of endothelium, mesangial cells is not observed. The epithelium of all departments of the tubular apparatus of the nephron is preserved. The vessels of the cortical and juxtamedullary sections of the kidney are moderately full-blooded. It should be noted the active functioning in the juxtamedullary departments of peritubular intercapillary anastomoses.

 Adrenal glands. The division of structures into the cortical and brain layers is not broken. In the cortical layer, the glomerular, bundle, and mesh zones are clearly differentiated. In the latter two, “voids” remain in the cytoplasm of cells, which was also observed in intact animals. Chromaffin cells of the brain layer are large, with well-contoured nuclei. Between them, sinusoidal capillaries are visible.

 Spleen. The trabecular vessels of the red pulp are full-blooded. They are surrounded by lymphocyte muffs. In the white pulp, the lymphoid follicles are clearly limited. Reactive centers are not expressed. Plasmatization was not found in them. Hyperplastic processes in the thymus-dependent and thymus-independent zones were not detected.

 Lungs. The epithelium in all subsections of the bronchial tree is preserved. The number of goblet cells in medium-caliber bronchioles is not increased. Gleams of terminal bronchioles, respiratory departments of the lungs are free. The interalveolar septa are thin. Gaps, emphysematous swelling in them were not found. The vessels of the interalveolar septa are moderately full-blooded.

 Heart. The fibrous components of the epicardium are thin. The transverse striation of myocardiocytes is clearly visible. Offsets of dark and light discs relative to each other, as well as with respect to the insert plates, were not detected. Intramuscular anastomoses are more distinctly revealed. Intramuscular connective tissue layers are thin without foci of basophilia. Arterioles, venules, capillaries are full-blooded. Gleams of their round shape without signs of manifest dilatation or dystonia.

 Thus, the two-month administration of “Karsilin” and “Karsil” in doses of 20 and 200 mg / kg did not reveal, judging by micromorphological data, destructive processes in the internal organs. Changes found in the liver (intracellular hyperplasia of hepatocytes, binuclear hepatocytes, activation of plastic metabolism in cells, hyperchromatosis of the nuclei), gastrohypermucoidization of the integumentary epithelium, the functioning of peritubular renal anastomoses, as well as intermuscular adaptations and prolonged cardiac arteriomas "Karsila" equally.

Claims (1)

Hepatoprotective agent, characterized in that it is made in tablet form and contains silymarin as an active substance in an amount of 10-20% by weight of the tablet core and target additives: microcrystalline cellulose, lactose, corn starch, crospovidone, polyvinylpyrrolidone, sodium bicarbonate, talc and stearic acid and / or its salts, while the core of the tablets is coated with a multilayer shell, the ingredients of which are from the total weight of the shell, wt.%:
I layer
Etazel - 0.05 - 0.4
Polyethylene glycol 6000 - 0.05 - 0.5
II layer
Sugar - 70.0 - 90.0
Polyvinylpyrrolidone - 0.2 - 0.6
Sodium carboxymethyl cellulose - 0.3 - 1.2
Twin 80 - 0.08 - 0.3
Talc - 5.0 - 9.0
Titanium Dioxide - 0.3 - 3.0
Aerosil - 0.15 - 0.5
III layer
Sugar - 4.0 - 9.0
Opalux AS 9840 Brown - 1.0 - 3.0
IV layer
Polyethylene glycol 6000 - 0.06 - 0.2
Glycerin - 0.005 - 0.03, and

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