RU2802015C1 - Method of obtaining a tissue preparation with hepatoprotective properties and an agent based on it - Google Patents

Abstract

FIELD: pharmaceutical industry.

SUBSTANCE: group of inventions relates to a method of producing a drug with hepatoprotective properties. A method of obtaining a tissue preparation with hepatoprotective properties includes taking the epidermis of the pelvic limbs of ducks, following freezing the raw material at a temperature of -20°C and defrosting at +22°C with rinsing under running water, followed double treatment with 0.05% chlorhexidine solution in the ratio of raw materials and chlorhexidine 1:4 by weight, and rinsing with 0.9% NaCl solution, after which drying is carried out in a dry-air cabinet at a temperature of 105°C to absolutely dry matter within 24 hours, the dry residue is ground in a mill at a speed of 14,000 rpm for 60 seconds to a particle size of 20–50 microns, the resulting powder is mixed with 0.9% NaCl solution in a ratio of 1:4, homogenized at a speed of 1,500 rpm for 60 seconds, poured into sterile vials, which are filled at 1/3, sterilized at a temperature of 120°C and a pressure of 1.1 atmospheres for 45 minutes. Tissue preparation with hepatoprotective properties obtained by the above method, contains the epidermis and 0.9% NaCl solution.

EFFECT: above method makes it possible to obtain a highly effective hepatoprotective agent from waterfowl processing waste.

2 cl, 6 dwg, 2 ex

Description

The field of technology to which the invention belongs

The present invention relates to veterinary medicine, specifically to methods and drugs for the treatment of animal hepatopathy.

A method of preparing a tissue preparation is known, in which skin and other organs from freshly killed animals are used. The skin is freed from subcutaneous fatty tissue, cut into small pieces weighing 2.5-20.0 g, washed from blood and placed for a day in a 2% solution of chloracid containing 400-500 mg of chlorine per liter. The chloracid solution is changed daily as it becomes discolored for 3-4 days and on the 6th day from the start of canning (Daricheva N.N., Ermolaev V.A. Tissue therapy in veterinary medicine: monograph. Ulyanovsk: UGSHA, 2011. 168 p. ). The disadvantage is that chloracid, according to the instructions, is intended for disinfection of surfaces from various materials of inanimate nature, with prolonged treatment of tissue preparations with it, it is impossible to completely wash them out of it.

A known method of preparing a tissue preparation from the mucous membrane and integumentary epithelium of the muscular stomach of chickens or ducks. Raw materials are taken from a bird at the age of 2-6 months during slaughter, treated with a saline solution containing chlorhexidine in a ratio of 20: 1 for 10-15 minutes, then washed with saline until the specific smell of chlorhexidine disappears, crushed, frozen at a temperature of -20- 30°C for 50-60 minutes, subjected to freeze drying and final drying (RU 2414917 C1). The disadvantage is that freeze-drying is an expensive technological process that does not provide sterility.

A known method of preparing a biogenic preparation from by-products and slaughterhouse waste (liver, spleen, lymph nodes, uterus with a fetus of 2-3 months). The raw materials are kept in a refrigerator for 5-7 days at a temperature of +2-+4°C, crushed, treated with a solution of sodium hypochlorite in the ratio of raw materials: sodium hypochlorite as 1:2-1:3, followed by extraction of raw materials in a refrigerator at a temperature +2-+4°С. Then, filtration, packaging and sterilization in an autoclave for an hour at a temperature of 120 ° C are carried out, extraction under refrigerator conditions is carried out for 8-10 hours, followed by dilution of the extract to a ratio of raw materials: sodium hypochlorite as 1: 4-1: 5 and extraction of the solution under the influence of ultrasound with a frequency of 20 kHz for 2.5-3 hours at a temperature not exceeding 50°C. The active solution of sodium hypochlorite is used at a concentration of 300.0-600.0 mg/l. The biogenic preparation obtained by this method is intended for subcutaneous and intramuscular administration to animals (RU 2698707 C1). The disadvantage is the duration and multi-stage process with a change in temperature.

The closest in technical essence to the claimed technical solution and taken as a prototype is a pharmaceutical oral composition intended for the treatment and prevention of recurrence of liver diseases caused by a violation of lipid-cholesterol metabolism, selected from the group: alcoholic and non-alcoholic steatohepatitis, primary biliary cirrhosis of the liver, cholesterosis gallbladder, drug and toxic damage to the liver, including as active principles a hepatoprotector from the group of bile acids or essential phospholipids, and as a prebiotic lactulose or fructooligosaccharides, taken in effective doses (EA019985 B1 (DIKOVSKY A.V.) 27.12.2014). The disadvantage is the complexity of manufacturing and demanding storage conditions.

Disclosure of invention

The objective of the invention is to develop a highly effective and safe agent for the prevention and treatment of hepatopathy, the manufacture of which contributes to the waste-free processing of waterfowl.

The technical result that can be achieved using the proposed invention is to obtain a highly effective hepatoprotective inexpensive agent from natural materials, waterfowl processing waste, which will lead to a reduction in the cost of treating animals with hepatopathy and an increase in the profitability of enterprises for growing and processing ducks.

The technical result is achieved by the fact that the method for obtaining a tissue preparation with hepatoprotective properties and an agent based on it includes disinfection and autoclaving at the final stage of preparation of the product, wherein the epidermis of the pelvic limbs of ducks, which is a waste product of the processing of waterfowl, is used as a raw material, additional freezing of raw materials is carried out for accumulation and storage at a temperature of -20°C, after which it is thawed at a temperature of 22°C and thoroughly washed under running water, followed by double treatment with 0.05% chlorhexidine solution and washing of raw materials with 0.9% NaCl solution, followed by drying in a dry-air cabinet at a temperature of 105°C until absolutely dry within 24 hours, the dry residue is ground in a laboratory mill at a speed of 14,000 rpm for 60 seconds, a powder with particles from 20 to 50 microns is obtained, containing 100 mg of the product 40.2% active ingredient. The powder is mixed with a 0.9% NaCl solution in a ratio of 1:4 and an additional 5% is added over the volume of distilled water (evaporates during autoclaving), homogenized in a homogenizer at a speed of 1500 rpm for 60 seconds, poured into sterile vials, which are filled 1/3, sterilized at a temperature of 120 ° C and a pressure of 1.1 atmospheres for 45 minutes, sealed with sterile rubber stoppers and run in with aluminum caps.

Under hepatopathy understand a group of liver diseases of various etiologies, characterized by toxic-inflammatory and degenerative damage to the liver parenchyma. Hepatopathy in animals and birds occur in different age periods and cause great damage to livestock and poultry. Therefore, the questions of finding and studying feed additives of a new generation, environmentally friendly and having hepatoprotective properties, are in demand.

The proposed drug is a suspension of powder of fine particles, 20-50 microns in size and 0.9% NaCl solution, bacteriologically safe, non-toxic (Fig. 1-3), with a shelf life of at least 1 year at a temperature of +4°C to +8°C, with hepatoprotective properties with the following characteristics in dry matter: moisture - no more than 2.03% and crude protein - 88.77%. Crude protein consists of 87.831% of amino acids, nitrogenous substances of non-protein origin account for 0.939% (Fig. 4), it also contains seventeen amino acids, seven of which are indispensable for mammals (valine, isoleucine, leucine, lysine, methionine, threonine, phenylalanine) and ten for birds (additionally arginine, histidine, tyrosine). All seventeen amino acids are involved in metabolic processes in the liver, six of which have pronounced antioxidant (glycine, alanine, cystine), detoxification (glutamic acid, threonine, arginine, methionine) and lipotropic (arginine, methionine) properties. The quantitative composition of amino acids in percentage terms in dry matter is as follows: aspartic acid (5.872), threonine (2.734), serine (8.255), glutamic acid (10.583), proline (4.467), glycine (11.546), alanine (3.017), cystine ( 2.133), valine (3.907), methionine (3.464), isoleucine (3.319), leucine (6.781), tyrosine (6.652), phenylalanine (4.271), histidine (1.111), lysine (3.049), arginine (6.676). Three trace elements (Cu, Zn, Mn) were found in dry matter, which play an important role in metabolic processes in the liver (Fig. 5). The combined effect of all the ingredients of the product and has a high hepatoprotective effect.

Brief description of the drawings

In FIG. 1- Particle sizes of the epidermis of the pelvic limbs of ducks after grinding to a powder state; uv. ×400.

In FIG. 2 - Biological safety test report.

In FIG. 3 - Results of chemical and toxicological analysis of the epidermis of the pelvic limbs of ducks.

In FIG. 4 - The amount of crude protein and amino acid composition of the studied samples.

In FIG. 5 - The content of trace elements in the dry matter of the epidermis.

In FIG. 6 - Biochemical parameters of blood of cats diagnosed with steatosis before treatment and on the sixth day after the start of treatment.

Implementation of the invention

The proposed method is carried out as follows. Examples of specific implementation of the proposed method.

Example No. 1. Raw materials are obtained from ducks aged forty or more days at the time of slaughter. Free from feather impurities, frozen at -20°C to accumulate material, thawed at 22°C, thoroughly washed under running water, placed in 0.05% chlorhexidine solution for 1 hour in the ratio of raw materials to chlorhexidine as 1:4 by weight . The bactericidal effect of chlorhexidine is manifested in a concentration of more than 0.01% at a temperature of 22 ° C and exposure for 1 minute, when ingested, it is practically not absorbed from the gastrointestinal tract (from 300 mg in 30 minutes, 0.206 μg / l can be absorbed in the digestive tract, refers to 4 class of low-hazard substances according to GOST 12.1.007-76, 90% is excreted by the intestines and less than 1% is excreted by the kidneys), change the chlorhexidine solution in the same ratio and incubate for another 30 minutes. Then the raw material is washed with a 0.9% NaCl solution, the excess liquid is allowed to drain on a metal sieve. In a dry-air cabinet, 1 kg of raw materials are laid out with a height of no whiter than 10 cm, dried at a temperature of 105 ° C to an absolutely dry substance for 24 hours, the dry residue is weighed on a laboratory electronic analytical balance VLTE-210 with a resolution of 0.0001 g manufactured by NPP Gosmetr , then the material is placed in a dry-air cabinet for 30 minutes, after which the sample is weighed again and drying is stopped at constant mass values. The dry residue is crushed in a laboratory mill at a speed of 14,000 rpm for 60 seconds; when packing, sterile vials of various volumes are filled by 1/3 on the basis that one dose corresponds to 0.6 g of powder and contains 240 mg of the active substance. Before use, 2.5 ml of 0.9% NaCl solution is added per dose, shaken thoroughly until a suspension is formed and administered orally (per os). The unused balance of the drug is stored in the refrigerator for two days. The resulting preparation is a friable powder of beige or light yellow color, well wetted, but insoluble in water, retains its color throughout the year. The disadvantage of this method is the native state of the protein, which slows down its absorption and the need to prepare a suspension before its use.

Example No. 2. Carried out similarly to example No. 1, but after grinding the dry residue in a laboratory mill, it is mixed with a 0.9% NaCl solution in a ratio of 1: 4, added to a volume of 5% distilled water to compensate for evaporation, homogenized in a single-phase homogenizer at a speed 1500 rpm for 60 seconds, pour into sterile vials depending on the number of doses based on the active ingredient per kilogram of animal weight (1 dose = 0.6 g of powder = 240 mg of active ingredient). The vials are filled to 1/3, sterilized by autoclaving at a temperature of 120 ° C and a pressure of 1.1 atmospheres for 45 minutes. The resulting tissue preparation is a suspension of light beige or light yellow color, ready for use, with a specified shelf life of 1 year.

The action of the proposed tissue preparation has been tested in veterinary clinics for the treatment of small animals on cats with established diagnoses of hepatosis (steatosis), which proved its positive effect on the dynamics of recovery during treatment. Animals with an average severity of the disease were prescribed complex treatment according to three schemes.

Scheme No. 1. Dietary feed, vitamins (groups B, D), macroelements (P, K, Mg), microelements (Zn, Cr), "Hepatovet" was used as a hepatoprotective agent according to the instructions.

Scheme No. 2. Dietary feed, vitamins (groups B, D), macroelements (P, K, Mg), microelements (Zn, Cr), as a hepatoprotective agent, the developed agent was used in the form of a powder, which was administered individually orally (per os) during feeding 2 times a day. Used powder packaged in 20 ml vials containing 4 doses for cats up to 4 kg.

Scheme No. 3. Dietary feed, vitamins (groups B, D), macroelements (P, K, Mg), microelements (Zn, Cr), as a hepatoprotective agent, the developed agent was used in the form of a suspension, which was administered individually orally (per os) during feeding 2 times a day, 3 ml (1 dose of 0.6 g of powder and 2.5 ml of 0.9% NaCl solution).

Biochemical parameters of blood before and after application of the tissue preparation are shown in the table (Fig. 6).

Claims (2)

1. A method for obtaining a tissue preparation with hepatoprotective properties, including taking the epidermis of the pelvic limbs of ducks, followed by freezing the raw material at a temperature of -20°C and defrosting at a temperature of +22°C with rinsing under running water, followed by a double treatment with a 0.05% chlorhexidine solution in the ratio of raw materials and chlorhexidine 1:4 by weight and washing with 0.9% NaCl solution, after which drying is carried out in a dry-air oven at a temperature of 105°C to an absolutely dry substance for 24 hours, the dry residue is ground in a mill at a speed of 14000 rpm min for 60 seconds to a particle size of 20-50 microns, the resulting powder is mixed with 0.9% NaCl solution in a ratio of 1:4, homogenized at a speed of 1500 rpm for 60 seconds, poured into sterile vials, which are filled for 1 /3, sterilized at a temperature of 120°C and a pressure of 1.1 atmospheres for 45 minutes. 2. Tissue preparation with hepatoprotective properties, obtained by the method according to p. 1, containing the epidermis and 0.9% NaCl solution.