RU2409380C1 - Method for preparing immunostimulating drug for animals and related immunostimulating drug - Google Patents

Abstract

FIELD: medicine, veterinary science. ^ SUBSTANCE: method for preparing an immunostimulating drug for animals consists in the fact that stevioside is dissolved in 0.9 % physiologic saline, the prepared solution is added to pre-grinded lecithin, then olive oil heated to 35-40C is introduced gradually in the prepared composition and mixed; further the prepared solution is filtered and autoclaved at temperature 115-120C for 15-20 minutes, then the produced sterile drug is bottled and corked up. The immunostimulating drug for animals contains stevioside, 0.9 % physiologic saline, olive oil, lecithin in a certain ratio. ^ EFFECT: higher viability and productivity of animals by correcting body immune responsiveness. ^ 2 cl, 4 tbl, 8 ex

Description

FIELD OF THE INVENTION:

The invention relates to veterinary medicine, in particular to immunostimulating preparations for animals and the method for their preparation, can be used to increase the general and specific resistance of the animal organism.

BACKGROUND

A plant-derived immunostimulant is known, amaranth seed oil. The proposed immunostimulant gives a clearly defined immunostimulating effect and can be used to correct immunodeficiency states in the treatment of diseases of various etiologies (see patent RU No. 2170096, class A61K 35/78, publ. 10.07.2001).

The disadvantage of this immunostimulant is the complexity of the preparation of the drug and the high consumption of materials.

A known method for producing a medicinal product based on a biologically active substance with hypoglycemic and immunomodulating activity, including extraction of plant materials, cuff grass with water, purified by the method of accelerated fractional maceration, in the ratio of raw materials: extractant - 1:10 when heated to 90-95 ° C ( see patent RU No. 2356569, class A61K 36/73, A61P 37/02, A61P 3/10, published on 05.27.2009).

The disadvantage of this method of obtaining an immunostimulant is the difficulty of obtaining the feedstock.

Known immunostimulant of plant origin, which proposes the use of (+) - R-licoricidin (1), a component of Ural licorice root, as well as dried powder and processed extract of Ural licorice root, containing 12-38 wt.% Substance (1) (see Patent RU No. 2166325, class A61K 35/78, A61K 31/58 publ. 05/10/2001).

The disadvantage of this immunostimulant is its short effect on the body.

The closest in technical essence and adopted by the authors for the prototype is a method for producing stevioside comprising the extraction of crushed dry leaves of a plant Stevia Rebaudiana B. liquefied CO ₂ in a subcritical or supercritical state. The invention provides a high degree of purification of the extract, reduces the loss of stevioside and increases the yield and purity of the finished product (see patent RU No. 2250041, class A23L 1/236, A23L 1/22, publ. 04/20/2005).

The disadvantage of this method is the powder form of the resulting substance, which makes it unsuitable for parenteral administration.

SUMMARY OF THE INVENTION

The objective of the invention is the creation of an immunostimulating drug based on stevioside, aimed at the treatment and prevention of immunosuppressive conditions caused by both pathological processes and an unfavorable environmental situation. Contributing to increasing the general and specific resistance of the body, the developed drug is a cheap and highly effective means for correcting the immune status of animals, thereby reducing the risk of death of animals from diseases of various etiologies.

The technical result that can be achieved using the present invention is to increase the viability and productivity of animals by correcting the immunological reactivity of the body.

The technical result is achieved using the method of obtaining an immunostimulating drug for animals, including the preparation of stevioside, while the stevioside is dissolved in 0.9% saline, the resulting solution is added to pre-ground lecithin, then olive oil is added to the resulting mixture, heated to 35-40 ° C. oil and mix, then filter the resulting solution and autoclave at a temperature of 115-120 ° C for 15-20 minutes, then the resulting sterile preparation is poured into a vial us and sealed.

The technical result is achieved using an immunostimulating drug for animals, including stevioside, additionally containing 0.9% saline, olive oil, lecithin, in the following ratio, wt.%:

Stevioside 10-30 Saline 0.9% 9-11 Lecithin 1-3 Olive oil rest

For the preparation of an immunostimulating drug for animals, stevioside was chosen as the main active substance. Stevioside is a diterpenoid glucoside with cardiotonic, hypoglycemic and immunostimulating effects on the animal organism. (Sehar I, and all. "Immune up regulatory response of a non-caloric natural sweetener, stevioside" Chem Biol Interact. 2008 May 28; 173 (2): 115-21. Epub 2008 Jan 31). Saline is needed to dissolve stevioside and prepare the emulsion. Lecithin is an affordable and effective emulsifier. Olive oil serves as a medium for the preparation of an immunostimulant and determines the prolonged effect of the drug on the animal organism.

The essence of the preparation of an immunostimulating drug for animals is as follows:

10, 20 and 30 (wt.%) Stevioside is dissolved in 9-11 (wt.%) 0.9% physiological saline, the resulting solution is added to lecithin 1-3, previously ground in a mortar, then up to 100% is gradually added to the resulting mixture olive oil heated to 35-40 ° C and mixed, then filter the resulting solution through a paper filter and autoclave at a temperature of 115-120 ° C for 15-20 minutes, the resulting sterile preparation is poured into vials and sealed, while the ratio of components to wt.% is:

Stevioside 10-30 Saline 0.9% 9-11 Lecithin 1-3 Olive oil rest

DETAILED DESCRIPTION OF THE INVENTION

Examples of specific performance of the method of preparation of an immunostimulating drug for animals and an immunostimulating drug based on it.

Example 1. 5 (wt.%) Stevioside is dissolved in 9 parts of a 0.9% saline solution, the resulting solution is added to the lecithin previously ground in a mortar, then olive oil heated to 20 ° C is gradually added to the resulting mixture and mixed, then filtered the resulting solution and autoclave at a temperature of 80 ° C for 10 minutes, the resulting preparation is poured into vials and sealed, while the ratio of components in wt.% is:

Stevioside 5 Saline 0.9% 9 Lecithin one Olive oil rest

When the drug was administered to animals, no reactions from the immune system were noted, oil heated to 20 ° C does not mix well with the other components, autoclaving at a temperature of 80 ° C for 10 minutes is not enough, since there were obvious signs of inflammation at the injection site.

Example 2. The drug is prepared analogously to example No. 1, but 7 (wt.%) Stevioside is dissolved in 9 milliliters of 0.9% saline, olive oil is heated to 25 ° C, autoclaved at a temperature of 90 for 15 minutes, while the ratio of components in wt.% is:

Stevioside 7 Saline 0.9% 9 Lecithin one Olive oil rest

When the drug was administered to animals, no immunostimulating effect was noted, the oil warmed up to 25 ° C, mixes very slowly with other components, autoclaving at a temperature of 90 for 15 minutes is not enough, since hyperemia occurred at the injection site.

Example 3. The drug is prepared analogously to example No. 2, but dissolve 10 (wt.%) Stevioside in 10 milliliters of 0.9% saline, olive oil is heated to 35 ° C, autoclaved at a temperature of 115 ° C for 15 minutes, while the ratio of components in wt.% is:

Stevioside 10 Saline 0.9% 10 Lecithin 2 Olive oil rest

A pronounced effect of the drug on the immune system is observed, the number of leukocytes increases, no negative effect on the body of experimental animals is noted. The drug due to the low concentration of stevioside is conveniently used for small animals (cats, dogs, rabbits, etc.).

Example 4. The drug is prepared analogously to example No. 3, but dissolve 20 (wt.%) Stevioside in 10 milliliters of 0.9% saline, olive oil is heated to 35 ° C, autoclaved at a temperature of 120 ° C for 15 minutes, while the ratio of components in wt.% is:

Stevioside twenty Saline 0.9% 10 Lecithin 2 Olive oil rest

This concentration of stevioside and the method of preparation of the drug is the most effective, since the most pronounced effect of the drug on the immune system is observed, and no negative effect on the body of experimental animals is noted. This concentration of stevioside in the preparation is convenient for use by medium-sized animals (sheep, goats, pigs, etc.).

Example 5. The drug is prepared analogously to example No. 4, but almond oil is used, while the ratio of components in wt.% Is:

Stevioside twenty Saline 0.9% 10 Lecithin 2 Almond oil rest

The high cost of the drug due to the scarcity and high cost of almond oil was noted.

Example 6. The drug is prepared analogously to example No. 4, but dissolve 30 (wt.%) Stevioside in 11 milliliters of 0.9% saline, olive oil is heated to 40 ° C, autoclaved at a temperature of 120 ° C for 20 minutes, while the ratio of components in wt.% is:

Stevioside thirty Saline 0.9% eleven Lecithin 3 Olive oil rest

An increase in the number of T- and B-lymphocytes against the background of the harmlessness of the drug on the animal organism indicates the effectiveness and this concentration of stevioside. This method of preparation is effective, but slightly prolonged. This concentration of stevioside should be used in large animals (cattle, horses, etc.).

Example 7. The drug is prepared analogously to example No. 6, but dissolve 40 (wt.%) Stevioside in 15 milliliters of 0.9% saline solution, olive oil is heated to 40-50 ° C, autoclaved at a temperature of 120-130 ° C for 25 -30 minutes, while the ratio of components in wt.% Is:

Stevioside 40 Saline 0.9% fifteen Lecithin four Olive oil rest

This concentration of stevioside caused a violation of biochemical processes and showed an undesirable effect on the immune system of the test animals. An increase in the proportion of saline solution increased the size of the dispersion phase in the emulsion. The decay rate of the emulsion did not decrease due to an increase in the proportion of emulsifier. An increase in temperature, autoclaving time and the proportion of emulsifier increased the cost of production.

Example 8. The drug is prepared analogously to example No. 7, but dissolve 50 (wt.%) Stevioside in 20 milliliters of 0.9% saline, olive oil is heated to 50-60 ° C, autoclaved at a temperature of 130-140 ° C for 30 -40 minutes, while the ratio of components in wt.% Is:

Stevioside fifty Saline 0.9% 25 Lecithin 5 Olive oil rest

Histological studies of the organs of experimental animals after the application of such a concentration of stevioside revealed degenerative changes in the liver. An obvious overspending of the emulsifier and physiological saline was noted, the emulsion contains large droplets of the dispersion phase and quickly decomposes into fractions. Time, autoclaving temperature and overspending of the emulsifier increases the cost of production.

ANIMAL IMMUNITY TEST RESULTS FOR ANIMALS

The developed drug was tested on sheep of 9-10 months of age of the Stavropol breed. The drug, based on a biologically active substance (stevioside), was administered intramuscularly to healthy sheep intramuscularly at a dose of 2 ml per 50 kg of live weight three times every three days. Blood for research from sheep was taken from the jugular vein on the 10th day after the last injection of the drug. The amount of hemoglobin, erythrocytes and leukocytes in the blood was determined, the level of total protein, albumin, α-β and γ-globulins, as well as indicators of immunological reactivity (the content of T and B lymphocytes, lysozyme, phagocytic and bactericidal activity) were determined in blood serum. . The test results of the drug based on stevioside are presented in tables 1, 2, 3 and 4.

Table 1 Hematological blood counts of sheep (M ± m) Group of animals Hemoglobin, g / l Red blood cells, × 10 ¹² / l White blood cells × 10 ⁹ / L Control 107.3 ± 0.6 8.6 ± 0.2 6.4 ± 0.3 Experienced 112.6 ± 1.7 8.7 ± 0.1 8.5 ± 0.3

Based on the data presented in table 1, an increase in the number of leukocytes by 33% (8.5 ± 0.3 × 10 ⁹ / L) in experimental animals was revealed, the number of hemoglobin and erythrocytes did not change significantly.

table 2 Biochemical blood parameters of sheep (M ± m) Group of animals Total protein, g / l Albumin, g / l Globulins, g / l α, g / l β, g / l γ, g / l Control 68.2 ± 1.2 16.9 ± 4.9 13.7 ± 1.8 12.7 ± 2.05 25.1 ± 5.4 Experienced 63.3 ± 1.7 12.3 ± 2.03 13.1 ± 2.9 8.7 ± 1.6 29 ± 4.03

From the data of table 2 it is seen that the immunostimulating drug for animals did not have a significant effect on such biochemical parameters as the amount of total protein and α-globulins. A 13% (29 ± 4.03 g / L) increase was observed relative to the control group of γ-globulins, against a background of a 30% decrease in the number of β-globulins (8.7 ± 1.6 g / L) and a 27% decrease in albumin (12, 3 ± 2.03 g / l).

Table 3 Immunological parameters of sheep blood (M ± m) Group of animals BASK,% FASK,% LASK,% T-lim.,% In-lim.,% Control 7.4 ± 0.4 34.6 ± 8.9 17.9 ± 0.5 60.2 ± 2.4 15.8 ± 0.6 Experienced 22.8 ± 1.1 40 ± 5.5 20.3 ± 1.2 79.6 ± 3.6 36.6 ± 3.7

Analyzing the immunological parameters presented in table 3, it should be noted the growth of bactericidal (BASK) and lysozyme (LASK) activity of blood serum of animals of the experimental group by 67% (22.8 ± 1.1%) and 11% (20.3 ± 1, 2%), respectively. The phagocytic activity of neutrophils (FASK) in relation to the control group increased by 13% (40 ± 5.5%). The relative number of T and B lymphocytes in the experimental group increased by 24% (79.6 ± 3.6%) and 56% (36.6 ± 3.7%), respectively.

Table 4 Productivity and preservation of animals (M ± m) Group of animals Live weight kg Incidence,% Case,% Getting offspring, goal. per 10 queens control 46 ± 2.5 13 ± 0.2 2.8 ± 0.7 11 ± 0.7 experienced 53 ± 1.6 8 ± 0.1 1.3 ± 0.4 13 ± 0.3

Analyzing the data presented in table 4, it should be noted that in the experimental group of animals the incidence decreased by 38% and the death of animals by 53%. After using the drug based on stevioside, the live weight of animals increased by 13% and the birth rate increased by 15%.

Based on the above research results, it is possible to recommend an immunostimulating drug for animals for use in veterinary medicine, as an immunostimulating agent that can act on specific and non-specific protective systems of the body.

The invention in comparison with the prototype and other known technical solutions has the following advantages:

- increase the viability and productivity of animals by correcting the immunological reactivity of the body;

- low cost and environmental friendliness of the drug;

- has a prolonged effect on the animal organism;

- the possibility of parenteral administration;

- various concentrations of the active substance.

Claims (2)

1. A method of obtaining an immunostimulating drug for animals from stevioside, while stevioside is dissolved in 0.9% physiological solution, the resulting solution is added to pre-ground lecithin, then olive oil heated to 35-40 ° C is gradually added to the resulting mixture and mixed then filter the resulting solution and autoclave at a temperature of 115-120 ° C for 15-20 minutes, then the resulting sterile preparation is poured into vials and sealed. 2. An immunostimulating drug for animals, including stevioside, characterized in that it additionally contains 0.9% saline, olive oil, lecithin in a ratio of components, wt.%:
Stevioside 10-30 Saline 0.9% 9-11 Lecithin 1-3 Olive oil rest