RU2278678C2 - Method for obtaining preparation to increase body resistance in animals - Google Patents

Abstract

FIELD: veterinary science.

SUBSTANCE: the present innovation deals with freezing the tissues of animal parenchymatous organs followed by defrosting, homogenization and hydrolysis. Hydrolysis should be conducted at the presence of succinic acid at pH being -4.0 and 45°C. Hydrolyzate should be decanted followed by addition of 3%-caustic soda solution up to pH=7.0; moreover, it is necessary to add novocain powder to hydrolyzate up to 0.4-0.5% concentration and gel of aluminium oxide hydrate at 2-3 mg/ml. The preparation obtained should be packed into 50-ml vials to be sterilized due to autoclaving at the mode of 1.0 atm. for 20 min. The innovation provides less reactogenic final product of prolonged action and increased anti-infectious activity.

EFFECT: higher efficiency.

2 ex, 2 tbl

Description

The method of obtaining the drug to increase the resistance of the animal organism.

The invention relates to veterinary medicine, in particular to a method for producing agents that increase the resistance of animals.

A known method of obtaining funds for increasing the resistance of the body of young animals, including cleaning the glandular tissue from connective tissue and fatty elements, washing them and homogenizing. Thymus, spleen and pancreas are used as glandular tissues, which are mixed after homogenization, add pancreas juice and preservative chinosol, and then the mixture is kept for 5-6 days at 38-40 ° С with periodic daily stirring (Aut. St. No. 904707, M. Cl. A 61 K 35/26, 1982. Means for increasing the body resistance of young animals and a method for its production, author A. M. Smirnov and others).

The disadvantage of this method is that the process of fermentation and hydrolysis of tissues using enzymes of the pancreas and gastric juice takes a long time (up to 6 days), and the temperature regime of 38-40 ° C favors the manifestation of growth activity of microflora, which requires the addition of a preservative. The hydrolyzate thus obtained contains a suspension of tissues, which substantially complicates the process of filtering through a gauze filter. To obtain a purified from balance substances hydrolyzate after filtration is subjected to centrifugation. The absence of a prolongator in the preparation shortens the period of its biological action in the body.

To eliminate the above disadvantages, a method for producing a drug that increases the resistance of an animal organism is proposed, including the use of known technological methods, in particular, freezing and thawing animal parenchymal organs, such as the spleen, liver, testicles, their homogenization, acid hydrolysis and purification with succinic acid.

What is new is that upon receipt of the tissue hydrolyzate, succinic acid, novocaine, and alumina hydrate gel are added to the homogenate. Succinic acid is itself a universal stimulator of metabolism of a living cell, a natural endogenous substrate of redox processes (Krebs cycle) in living organisms.

The addition of novocaine is due to the need to relieve pain during the parenteral administration. In addition, tissue preparations prepared on a solution of novocaine have a more rapidly stimulating effect than suspensions prepared on a physiological solution.

Adding alumina hydrate to the gel hydrolyzate allows for a longer period of action, and therefore there is no need to conduct a course of frequent use of the drug.

The result is a less reactogenic drug with a pronounced anti-infectious effect.

Example 1. Obtaining a drug to increase body resistance using spleen tissue.

To obtain the drug used 500 g of the spleen, which was subjected to freezing in the freezer. After partial thawing, they were homogenized. 5 parts of distilled water were added to the obtained homogenizate, in which succinic acid was previously dissolved to pH 4.0. The resulting mixture was thoroughly mixed and placed in a thermostat. Acid hydrolysis was carried out at 45 ° C. After complete separation of the homogenate into the liquid hydrolyzed part and the precipitate from the non-hydrolyzed part of the tissues, the hydrolyzate was decanted. By fractional addition of a 3% sodium hydroxide solution, the pH was adjusted to a neutral pH of 7.0.

The required amount of novocaine powder was calculated so that its concentration in the hydrolyzate was in the range of 0.4-0.5%.

To sorb the biologically active substances of the preparation, an alumina hydrate gel was added to the hydrolyzate at a rate of 2-3 mg / ml. The resulting hydrolyzate was packaged in 50 ml vials and sterilized by autoclaving at 1.0 atm for 20 minutes.

The sterility of the drug was evaluated according to the results of the control culture of the contents of the vials on meat peptone agar and meat peptone broth. No growth of microorganisms was observed, indicating the sterility of the hydrolyzate.

The anti-infectious activity of the indicated spleen hydrolyzate was tested in modeling an acute infectious process in white mice.

The comparative evaluation procedure included a single intraperitoneal administration of splenic hydrolysates at a dose of 0.5 ml to white mice weighing 20-21 g, followed by their infection with E. coli culture at a dose of 4 LD ₅₀ . Infection was carried out one day later, on the 7th and 14th day after immunization. The results of the observations are presented in table 1.

Table 1.
Anti-infectious activity of spleen hydrolysates in modeling an acute infectious process with E. coli culture in white mice. A drug Infection of mice later 12 hours 7 days 14 days Hydrolyzate with Novocaine 10/4 10/4 10/5 Hydrolyzate with Alumina Hydrate Gel Additive 10/4 10/4 10/3 0.85% sodium chloride solution (control) 3/3 3/3 3/3 Note: the numerator is the number of infected mice; in the denominator - from the number of infected fell.

Thus, the addition of alumina hydrate to the gel hydrolyzate made it possible to stabilize the anti-infective activity of the drug in long periods after immunization of mice.

Example 2. Obtaining a drug to increase the body's resistance using liver tissue.

To obtain the drug used 300 g of the liver. Liver tissue was frozen in a freezer. After partial thawing, it was homogenized. 5 parts of distilled water were added to the obtained homogenate, in which succinic acid was dissolved to pH 4.0. The resulting mixture was thoroughly mixed. Acid hydrolysis was carried out at a temperature of 45 ° C. After 3 hours, there was a complete separation of the contents of the bio-bottle with the homogenate into the liquid part of the hydrolyzate and sediment. By decantation, the hydrolyzate was separated from the precipitate. By adding a 3% sodium hydroxide solution, the increased acidity of the hydrolyzate was neutralized. An alumina hydrate gel of 2-3 mg / ml was added to the hydrolyzate.

Packing, sterilization and control on the sterility of the hydrolyzate was carried out, as in example 1.

The anti-infectious activity of the hepatic hydrolyzate was tested on white mice weighing 17-18 g when simulating an acute infection process with a S. aureus culture at a dose of 4LD ₅₀ . The results of the observations are presented in table 2.

Table 2.
Anti-infectious activity of liver hydrolysates in modeling an acute infectious process by S. aureus culture in white mice. A drug Infection of mice later 12 hours 7 days 14 days Hydrolyzate with Novocaine 7/4 7/4 7/6 Hydrolyzate with Alumina Hydrate 7/4 7/3 7/4 The hydrolyzate without the addition of novocaine and aluminum oxide hydrate 7/4 7/4 7/6 0.85% sodium chloride solution (control) 3/3 3/3 3/3 Note: numerator - the number of infected mice; the denominator is the number of fallen mice.

Based on the second series of experiments on testing the amber hydrolyzate of the liver, it was found that the addition of alumina hydrate to the gel preparation prolongs the anti-infection activity of the hydrolyzate in the long-term period after immunization.

The lower level of anti-infectious activity of the liver hydrolyzate with respect to the spleen hydrolyzate is due to the fact that the latter refers to the organs of the immune system, which has a more pronounced immunogenic effect.

Claims (1)

A method of obtaining a drug to increase the resistance of an animal organism, including freezing tissues of parenchymal organs, thawing, homogenization, hydrolysis, characterized in that the hydrolysis is carried out in the presence of succinic acid at a pH of 4.0 at 45 ° C, the hydrolyzate is decanted, the pH of the hydrolyzate is adjusted to 7 , 0, novocaine powder is dissolved in the hydrolyzate to its concentration in the preparation equal to 0.4-0.5%, and an alumina hydrate gel is added at a rate of 2-3 mg / ml.