RU2770534C1 - Hepatoprotective and antioxidant drug for animals based on aqueous solution of fullerene c60, resveratrol and betaine hydrochloride - Google Patents

Abstract

FIELD: medicine; veterinary pharmacy.

SUBSTANCE: present invention relates to the field of medicine, namely to veterinary pharmacy, and it is a hepatoprotective and antioxidant drug for animals based on an aqueous solution of fullerene C₆₀, resveratrol and betaine hydrochloride, at the following ratio of components, mg: aqueous solution of fullerene C₆₀ stabilized with pluronic F-127 – 1 ml, 1 mg of active substance; resveratrol – 25 mg; betaine hydrochloride – 10 mg.

EFFECT: stimulation of liver protection functions and antioxidant effect, inhibition of free-radical processes in the animal organism, increase in the stress resistance of the organism, reduction in toxicity, increase in the convenience of use and dosage.

1 cl, 3 ex, 10 tbl, 8 dwg

Description

The technical field to which the invention belongs

The invention relates to veterinary pharmacy, in particular to preparations for the prevention and treatment of liver diseases, free radical pathology in animals and can be used in veterinary medicine.

State of the art

Known connection of an aqueous solution of fullerene enC ₆₀ . It has been established that an aqueous solution of fullerene dnC ₆₀ exhibits pronounced virucidal, prophylactic and therapeutic activity. The therapeutic effect in vivo was tested on a model of cutaneous herpetic infection in mice. As a result of treatment of animals with an aqueous solution of fullerene dnC ₆₀ , a statistically significant decrease in the clinical manifestations of herpes infection was observed. The results obtained allow us to conclude that the aqueous solution exhibits a pronounced therapeutic activity against HSV-1 infection, while its therapeutic effect is comparable to the reference drug ACV, when using 500 times lower concentration (See US Pat. No. RU 2694754 C1, publ. 07/16/2019).

The disadvantage of the compound aqueous solution of fullerene dnC ₆₀ is the lack of hepatoprotective and antioxidant effects. Not used in veterinary pharmacology.

A known method of obtaining a vaccine containing fullerene With ₆₀ -ser as an adjuvant. At the same time, a significant (adjuvant) immunostimulating effect of fullerene in relation to antigenic extracts from bacterial biomass is provided. The method involves the treatment of an antigenic extract from the biomass of a microbe with sodium periodate at a concentration of 0.5-1.5 mg/ml in terms of total protein content and a sodium periodate concentration of 20-120 mmol/ml in the reagent mixture at a pH of 5.0-6.0 . Fullerene C ₆₀ -ser is used as an adjuvant. (See patent RU 2184566 C1, publ. 10.07.2002).

The disadvantage of the method for obtaining a vaccine containing fullerene With ₆₀ -ser, as an adjuvant as a drug is the lack of hepatoprotective and antioxidant effects. Not used in veterinary pharmacology.

It is known to use water-soluble fullerene derivatives as neuroprotective and antitumor drugs (See Pat. No. RU 2704483 C2, publ. 29.10.2019).

The disadvantage of water-soluble derivatives of fullerene compounds as neuroprotective and antitumor drugs is the lack of hepatoprotective and antioxidant effects. Not used in veterinary pharmacology.

Known fullerene derivative of glycopeptide formula I: Glc Nac-Mur Nac-Ala-DiGln[Lys-ε-CO(CH ₂ )5NHC60-H], with adjuvant activity. Obtained by synthesis of the fullerene derivative of aminocaproic acid and its subsequent addition to the lysine derivative of the glycopeptide. Experiments carried out on a group of mice (female Balb/c line) showed that at low doses of 1-10 nm/animal, the synthesized compound has high adjuvant properties (See Pat. No. RU 2124022 C1, publ. 27.12.1998) .

The disadvantage of the fullerene derivative of the glycopeptide of formula I: Glc Nac-Mur Nac-Ala-DiGln[Lys-ε-CO(CH ₂ )5NHC60-H], is the lack of hepatoprotective and antioxidant effects. Not used in veterinary pharmacology.

Disclosure of invention

The objective of the invention is to develop a highly effective drug based on an aqueous solution of C ₆₀ fullerene, which has pronounced hepatoprotective and antioxidant, immunostimulating effects, low toxicity, high efficiency in the prevention and treatment of liver pathologies, free radical pathologies in animals, ease of administration and dosage.

The technical result that can be achieved using the present invention is to stimulate the protective functions of the liver and the antioxidant effect, inhibition of free radical processes in the animal body, increase the stress resistance of the body, reduce toxicity, improve ease of use and dosing.

The technical result is achieved using a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride, including an aqueous solution of C ₆₀ Fullerene, resveratrol and betaine hydrochloride, with the following ratio of components in wt. mg:

Aqueous solution of C ₆₀ fullerene, stabilized with F-127 pluronic - 1 ml, 1 mg according to the active substance.

Resveratrol - 25 mg.

Betoin hydrochloride - 10 mg.

The claimed hepatoprotective and antioxidant drug for animals based on an aqueous solution of fullerene C ₆₀ , resveratrol and betaine hydrochloride is a transparent solution of dark brown color, odorless, with a pronounced hepatoprotective, antioxidant, antihypoxic effect, is an inhibitor of free radical processes, increases the body's stress resistance, low toxicity and easy to administer and dose.

As a vital organ, the liver performs a wide range of functions in the body, including detoxification, plasma protein synthesis, and glycogen storage. Oxidative stress is seen as an imbalance between the production of reactive oxygen species (ROS) and the antioxidant defense mechanism. This is the main reason for the development of liver diseases (Kseiko D.A. Antioxidant protein system in the liver in acute blood loss / Ulyanovsk biomedical journal. - 2012. - No. 1. - P. 64-67). Various types of liver diseases are characterized by cirrhosis, jaundice, tumors, metabolic and degenerative lesions, necrosis of liver cells, etc. Treatment of liver diseases is still a problem for modern veterinary medicine (Reshetnyak V.I., Maev I.V., Reshetnyak T.M., Zhuravel S.V., Pisarev V.M. Liver diseases and hemostasis (review) part 1. Non-cholestatic liver diseases and hemostasis - General resuscitation, 2019. - V. 15. - No. 5. - P. 74-87.).

Oxidative stress is one of the most common toxic factors. All types of agents (chemical, physical and biological) can induce oxidative stress in tissues and cells. Moreover, oxidation-mediated reactions are involved in many fundamental aspects of life processes, such as cellular respiration (mitochondria), lipid synthesis, metal metabolism, lysosomes, immunity, and xenobiotic biotransformation of organic compounds (Skripnichenko E.V. Effect of physical activity on redox processes in the body of an athlete: "Physical education and sport in higher educational institutions", collection of articles of the XVI International scientific conference - Belgorod, 2020. - P. 139-144.).

The liver is also affected due to the damaging effects of free radicals and leads to fatty changes in the liver. It is well known that normal liver function is essential for maintaining CNS health. Altered metabolic products accumulated due to hepatocyte dysfunction can cause brain damage and contribute to damage to the central nervous system (CNS). It is important to note that liver failure caused by various factors causes significant changes in neuron-supporting cells called astrocytes. Astrocytes are the most common cell type present in the CNS. They provide metabolic and trophic support for nerve cells, maintain the blood-brain barrier, modulate synaptic activity and protect neurons from damage (Arutyunyan A.V., Milyutina Yu.P., Zaloznyaya I.V., Shcherbitskaya A.D., Kerkeshko G.O. The role of placental neurotrophic factors in fetal brain development in hyperhomocysteinemia - Molecular Medicine - 2019. - V. 17. - No. 3. - P. 3-7.

The release of reactive oxygen species during these processes is neutralized by a suite of antioxidant compounds in an attempt to protect the integrity of cells or tissues.

The delicate balance between antioxidants and reactive oxygen species production can be disturbed by exposure to toxic substances.

Many reviews have been devoted to the antioxidant properties of fullerenes.

Fullerenes are a relatively new group of compounds and are a class of spherical molecules composed entirely of carbon atoms.

Fullerene has two main properties, for which it can be called a two-faced Janus - the ability to neutralize free radicals, acting as an antioxidant, and the ability to generate reactive oxygen species when irradiated (Piotrovsky L.B., Litasova E.V., Dumpis M.A. Piotrovsky L.B., Kiselev O.I. Why do we need fullerenes today? - 2019. - V. 17. - No. 2. - P. 5-15. Fullerenes in biology - St. Petersburg: Rostock, 2006. - 334 p.). It is the ability to neutralize free radicals, to act as a trap, that is, to exhibit the properties of an antioxidant, that is the main property of fullerene, which determines its demand in various areas of the national economy. For the first time, the ability to trap free radicals, that is, the antioxidant effect of fullerene, was discovered by Krusic et al. (Krusic PJ, Wasserman E, Keizer PN, et al. Radical reaction of C _60. Science. 1991;254:1183-1185. https://doi.org/10.1126/science.254.5035.1183). They even called the C60 _fullerene a "radical sponge" because it trap 34 methyl or 15 benzyl radicals. The addition of radicals to C ₆₀ fullerene occurs at the C=C double bond to form the corresponding adducts (McEwen CN, McKay RG, Larsen BS. C ₆₀ as a radical sponge. J Am Chem Soc. 1992;114(11):4412-4414. https://doi.org/10.1021/ja00037a064), which is the chemical basis for the ability of fullerenes to exhibit the properties of antioxidants and cytoprotectors in biological systems (Yin JJ, Lao F, Fu PP, et al. The scavenging of reactive oxygen species and the potential for cell protection by functionalized fullerene materials Biomaterials 2009;30(4):611-621 https://doi.org/10.1016/j.biomaterials.2008.09.061).

The essence of the obtained hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride, in the following ratio of components in wt. mg:

Aqueous solution of C ₆₀ fullerene, stabilized with F-127 pluronic - 1 ml, 1 mg according to the active substance.

Resveratrol - 25 mg.

Betoin hydrochloride - 10 mg.

The solution is mixed under aseptic conditions and packaged.

Implementation of the invention

Examples of a specific implementation of obtaining and testing using a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride.

Example 1

Hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride, with the following ratio of components in wt. mg:

Aqueous solution of C ₆₀ fullerene, stabilized with F-127 pluronic - 1 ml, 1 mg according to the active substance.

Resveratrol - 25 mg.

Betoin hydrochloride - 10 mg.

The resulting solution had a significant impact on the processes of lipid peroxidation and the state of the antioxidant system. The concentration of diene conjugates in the blood serum decreased by 49.8% (Fig. 1).

The concentration of malondialdehyde (MDA) decreased in the blood serum - by 38.8%, in the brain tissue - by 39.4%, in the lungs - by 18.2%, in the kidneys - by 18.1%, in the liver - by 24, 8%, heart - by 14.9%, skeletal muscles - by 12.4%, in the stomach - by 17.6%, in the small intestine - by 19.5% and in the large intestine - by 20.4% (Table . one).

Catalase activity after administration of the drug based on an aqueous solution of fullerene C ₆₀ significantly increased in blood serum by 7.5%, in brain tissue by 10.4%, in lung tissue by 9.7%, in gastric tissue by 11%, 8% and tissues of the small intestine - by 7.6%. In other tissues, the enzyme activity was within the physiological norm (Table 1).

Example 2

Hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride, with the following ratio of components in wt. mg:

Aqueous solution of C ₆₀ fullerene, stabilized with F-127 pluronic - 1 ml, 1 mg according to the active substance.

Resveratrol - 25 mg.

Betoin hydrochloride - 10 mg.

The resulting solution had a significant impact on the state of protein-nitrogen metabolism and urea-forming function of the liver, which was assessed by the dynamics of total protein in the blood, transamination enzymes, as well as metabolites of the ornithine cycle and the glutamic acid-glutamine system (Table 2). The drug was administered subcutaneously to white rats.

It was found that after the introduction of the studied compound, the content of total protein, urea, ammonia and ornithine increased by 8.6%, 10.0%, 10.0%) and 11.7%, respectively, compared with healthy ones (Table 2). There were no significant differences in the content of glutamine in the blood serum before and after the administration of the studied compound.

Urea and glutamine play an important role in acid-base regulation. Renal production of ammonia from glutamine is a well-known response to metabolic acidosis. Since, along with urinary ammonium excretion, glutamine synthesis is the second main mechanism by which mammals can absorb excess ammonia.

After the introduction of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride, the activity of transamination enzymes also increases (Fig. 2, 3).

Tranaminases are usually released at a constant rate, with normal levels in healthy animals representing a balance between normal hepatocyte turnover due to programmed cell death (apoptosis) and plasma clearance of enzymes.

Alanine aminotransferase (ALT) is present only in the cytoplasm of hepatocytes, while aspartate aminotransferase (ACT) is present in both the cytoplasm of hepatocytes and mitochondria. The functions of both of these transaminases represent important metabolic links between carbohydrate and protein metabolism. ALT is involved in the "glucose-alanine cycle" and exchanges alanine and pyruvate and can regenerate glucose consumed by the muscles in the same way that lactate dehydrogenase exchanges lactate and pyruvate in the "Cori cycle" to regenerate glucose from lactate in anaerobic metabolism.

It is very significant that in animals the content of ornithine in the blood after the introduction of the studied compound increased by 11.7%. Since ornithine helps to remove two harmful gases from the body, ammonia and carbon dioxide. Ammonia is highly toxic to the central nervous system and must be eliminated from the body. The stages of this cycle occur in the mitochondria and cytoplasm.

Our results indicate that after the introduction of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride, arginase activity increases by 20.7% compared to control animals (Fig. 4).

As for the activity of lactate dehydrogenase (Fig. 5), after the introduction of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride, it did not change significantly.

It was found that the concentration of total protein after the introduction of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride increased by 8.6% compared to control animals (Table 3). Other studied indicators of protein metabolism changed in different ways. Thus, the content of albumin in the blood increased by 11.5%. At the same time, the concentration of α-globulins after the introduction of the studied compound did not change significantly, β-globulins increased by 10.7% and γ-globulins - by 16.7% relative to the control.

Thus, a hepatoprotective and antioxidant drug for animals based on an aqueous solution of fullerene C ₆₀ , resveratrol and betaine hydrochloride, with the following ratio of components in wt. mg: an aqueous solution of C ₆₀ fullerene, stabilized with pluronic F-127 - 1 ml, 1 mg of the active substance, resveratrol - 25 mg, betoin hydrochloride - 10 mg, has a pronounced hepatoprotective effect.

Example 3

Acute toxicity of a hepatoprotective and antioxidant preparation for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride (24 animals) and white rats (24 animals).

In white rats, the drug was administered subcutaneously in doses of 550, 825, 1100, 2500, 5000 mg/kg of the active substance, in white mice - intragastrically in the same doses. The animals were observed for 14 days after drug administration. The volume of the injected solution was up to 1.0 ml for white rats and up to 0.5 ml for white mice. The research results are shown in tables 4 and 5.

Clinical toxicity after subcutaneous administration of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride in white rats was accompanied by the following symptoms (Table 6).

Animals treated with a hepatoprotective and antioxidant drug for animals based on an aqueous solution of fullerene C ₆₀ , resveratrol and betaine hydrochloride in the studied doses during the entire observation period (14 days) actively moved around the cages, in terms of eating and drinking behavior differed little from rats in the control group, adequate reaction to external stimuli (sound, light) was at the control level.

With intragastric administration of the drug at a dose of 2500 mg/kg, white mice were depressed, inactive, and refused food during the first day. Reaction to external stimuli was reduced. On the second day after the administration of the drug, eating and drinking behavior, reactions to external stimuli were restored (Table 7).

Within 14 days of research, the death of experimental animals was not noted. In this regard, it was not possible to install LD ₅₀ . This allows us to consider a hepatoprotective and antioxidant drug for animals based on an aqueous solution of fullerene C ₆₀ , resveratrol and betaine hydrochloride, non-toxic in the tested range and vary its doses over a wide range.

Example 3

The irritating effect of the hepatoprotective and antioxidant drug for animals based on an aqueous solution of fullerene C ₆₀ , resveratrol and betaine hydrochloride was evaluated on guinea pigs and rabbits based on the results of the method of skin applications and conjunctival tests.

Guinea pigs and rabbits were used in several batches to establish the skin test method for a hepatoprotective and antioxidant preparation for animals based on an aqueous solution of _С60 fullerene, resveratrol, and betaine hydrochloride. The irritating effect of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride was evaluated on guinea pigs in the amount of 10 animals divided into 2 groups.

Guinea pigs of the 1st experimental group were treated with a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride, with the following ratio of components in wt. mg: aqueous solution of C ₆₀ fullerene, stabilized with pluronic F-127 - 1 ml, 1 mg of active ingredient, resveratrol - 25 mg, betoin hydrochloride - 10 mg.

Guinea pigs of the 2nd experimental group were injected with a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride. The solution is prepared by mixing in 1 ml of an aqueous solution of Fullerene C ₆₀ 1 ml, 1 mg of the active ingredient, 2500 mg of resveratrol, 1000 mg of betoin hydrochloride.

A hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol, and betaine hydrochloride was applied to the skin. The injection site was the right lateral surface of the middle of the body. Distilled water was applied to the left lateral surface of the body as a control. Preliminarily, a body area measuring 2 cm × 2 cm was cut short. Solutions of the drug based on an aqueous solution of C ₆₀ fullerene for the prevention and treatment of free radical pathology in animals and distilled water were applied 3 drops once a day for 14 days. To exclude the development of contact dermatitis, the mechanical effect on the experimental skin area was minimized (Fig. 6, 7).

The reaction of the skin of guinea pigs during the cutaneous application of aqueous solutions of C ₆₀ fullerene was assessed according to the Patterson Skin Test Scale (Table 8). After applying the drug, the animals were continuously monitored for 20 minutes. Then the test values were taken every 24 hours. During the experiments, it was found that a hepatoprotective and antioxidant drug for animals based on an aqueous solution of fullerene C ₆₀ , resveratrol and betaine hydrochloride does not irritate the skin of experimental guinea pigs.

The irritating effect of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of fullerene C ₆₀ , resveratrol and betaine hydrochloride was studied in 2 series of experiments. For this, guinea pigs were used in the amount of 10 heads, divided into 2 groups.

Guinea pigs of the 1st experimental group were treated with a hepatoprotective and antioxidant preparation for animals based on an aqueous solution of fullerene C ₆₀ , resveratrol and betaine hydrochloride, with the following ratio of components in May. mg: aqueous solution of fullerene C ₆₀ stabilized with pluronic F-127 - 1 ml, 1 mg of active substance, resveratrol - 25 mg, betoin hydrochloride - 10 mg (Table 9).

Guinea pigs of the 2nd experimental group were injected with a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride. The solution is prepared by mixing in 1 ml of an aqueous solution of Fullerene C ₆₀ - 1 ml, 1 mg of the active substance, 2500 mg of resveratrol, 1000 mg of betoin hydrochloride.

Distilled water was applied to the surface of the back on the right side to take control data.

Daily monitoring of the skin condition at the sites of application of the drug found that the studied drug in the 1st and 2nd form of aqueous solutions does not cause persistent morphological changes in different skin layers in guinea pigs over the entire observation period.

Similar experiments on rabbits were carried out with a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol, and betaine hydrochloride. For this purpose, 2 groups of rabbits were selected, 6 heads each.

Rabbits of the 1st experimental group used a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride, with the following ratio of components in wt. mg: aqueous solution of fullerene C ₆₀ stabilized with pluronic F-127 - 1 ml, 1 mg of active ingredient, resveratrol - 25 mg, betoin hydrochloride - 10 mg (Table 10).

Rabbits of the 2nd experimental group were injected with a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride. The solution is prepared by mixing in 1 ml of an aqueous solution of Fullerene C ₆₀ - 1 ml, 1 mg of the active substance, 2500 mg of resveratrol, 1000 mg of betoin hydrochloride.

It was established that application of solutions 1 and 2 to the skin did not have a pathological effect on the skin of rabbits both on the 7th and 14th days of the experiments.

The conducted studies on the assessment of the irritating effect on the skin of animals of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride allow us to conclude that the drug based on an aqueous solution of C ₆₀ fullerene for the prevention and treatment of free radical pathology in animals does not cause pronounced morphological changes in different layers of the skin of experimental animals.

Guinea pigs were used to perform the conjunctival test. Guinea pigs of the 1st experimental group were treated with a hepatoprotective and antioxidant preparation for animals based on an aqueous solution of fullerene C ₆₀ , resveratrol and betaine hydrochloride, with the following ratio of components in May. mg: aqueous solution of C ₆₀ fullerene, stabilized with pluronic F-127 - 1 ml, 1 mg of active ingredient, resveratrol - 25 mg, betoin hydrochloride - 10 mg.

Guinea pigs of the 2nd experimental group were injected with a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride. The solution is prepared by mixing in 1 ml of an aqueous solution of Fullerene C ₆₀ - 1 ml, 1 mg of the active substance, 2500 mg of resveratrol, 1000 mg of betoin hydrochloride.

Ready-made solutions, 1 drop, were injected with an eye pipette under the upper eyelid of the right eye, and the left eye served as a control, and distilled water was injected. To assess the allergenic effect of each drug, 10 guinea pigs, 5 animals in groups, were used.

Immediate-type hypersensitivity was assessed 15 minutes after drug administration. Delayed type hypersensitivity was assessed after 24-48 hours. The results of the studies (Table 8) were evaluated according to the following scale:

1 - slight reddening of the lacrimal duct;

2 - reddening of the lacrimal duct and sclera towards the cornea;

3 - redness of the entire conjunctiva and sclera. The reaction is accompanied by itching and when combing with the paws, the development of purulent ophthalmitis is possible.

Within 6 hours of continuous observation of animals, there were no changes in the clinical state of the organism of guinea pigs after application of the drug to the conjunctiva (Fig. 9). According to the methodology, after a 24-hour and 48-hour observation period, no changes were recorded in the state of guinea pigs either.

Brief description of drawings and other materials

A graphic representation of the content of diene conjugates in the blood serum of white rats after subcutaneous administration of a hepatoprotective and antioxidant preparation for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride is shown in Fig. one.

The concentration of peroxidation products in the blood of animals and the activity of antioxidant enzymes in the blood of animals are presented in Table 1.

Indicators of protein-nitrogen metabolism in the blood of white rats are presented in Table 2.

A graphical representation of aspartate aminotransferase activity, (u/l), in the blood serum of white rats after subcutaneous administration of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride is shown in figure 2.

A graphic representation of the activity of alanine aminotransferase, (u/l), in the blood serum of white rats after subcutaneous administration of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride is shown in figure 3.

A graphical representation of the activity of arginase, (μmol/urea/g of raw tissue), in the blood serum of white rats after subcutaneous administration of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride is shown in figure 4.

A graphical representation of the activity of lactate dehydrogenase, (mmol/g), in the blood serum of white rats after subcutaneous administration of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride is shown in figure 5.

Indicators of protein metabolism in the blood of white rats are presented in Table 3.

Acute toxicity for white rats of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride is presented in Table 4.

Acute toxicity for white mice of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride is presented in Table 5.

The state of the physiological parameters of white rats with a single subcutaneous injection of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride at a dose of 2500 mg/kg for the active substance is presented in table 6.

The state of the physiological parameters of white mice with a single intragastric administration of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride at a dose of 2500 mg/kg for the active substance is presented in table 7.

In FIG. 6, 7 - illustrations of the skin-resorptive reaction for guinea pigs of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride.

The rating scale for application skin tests is presented in Table 8.

An assessment of the irritating effect of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride for guinea pigs and rabbits is presented in Table 9.

The results of conjunctival tests of a hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride for rabbits are presented in Table 10.

In FIG. Figure 8 illustrates an allergic reaction for guinea pigs to the administration of a hepatoprotective and antioxidant preparation for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol, and betaine hydrochloride.

Claims (4)

Hepatoprotective and antioxidant drug for animals based on an aqueous solution of C ₆₀ fullerene, resveratrol and betaine hydrochloride with the following ratio of components in mg: Aqueous solution of C ₆₀ fullerene, stabilized with F-127 pluronic - 1 ml, 1 mg according to the active substance; Resveratrol - 25 mg; Betoin hydrochloride - 10 mg.

Images