Method for preparing immunostimulating drug for animals and related immunostimulating drug

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 35-40°C is introduced gradually in the prepared composition and mixed; further the prepared solution is filtered and autoclaved at temperature 115-120°C 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

 

The technical FIELD TO WHICH the INVENTION RELATES:

The invention relates to veterinary medicine, in particular to the immune system, drugs for animals and method of production thereof, can be used to improve the overall and specific resistance of the organism of animals.

The LEVEL of TECHNOLOGY

Known immunostimulatory agent of vegetable origin, oils from seeds of amaranth. The proposed immune gives clearly marked immunostimulating effect and can be used for the correction of immunodeficiency in the treatment of diseases of different etiology (see patent RU No. 2170096, CL A61K 35/78, published. 10.07.2001).

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

A method of obtaining medicines on the basis of biologically active compounds having hypoglycemic and immunomodulatory activity, including the extraction of vegetable raw materials, herbs cuff with water, purified by the method of accelerated fractional maceration, in the ratio of raw material:extractant - 1:10 when heated to 90-95°C. (see patent RU No. 2356569, CL A61K 36/73, OR 37/02, OR 3/10, publ. 27.05.2009).

The disadvantage of this method of obtaining immunostimulant is the difficulty of obtaining raw material.

Known immunostimulant herbal about the convergence, where the use of (+)-R - limitedin (1), a component of licorice root Ural, and dried in powder form and processed licorice root extract Ural containing 12-38 wt.% substance (1) (see patent RU No. 2166325, CL A61K 35/78, A61K 31/58 publ. 10.05.2001).

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

The closest in technical essence and adopted by the authors for the prototype is a way to get the reveal includes the extraction of crushed dried leaves of the plant Stevia Rebaudiana Century liquefied CO2located in a subcritical or supercritical condition. The invention provides a high degree of purification of the extract, reduces loss of reveal and increases the yield and purity of the finished product (see patent RU No. 2250041, CL A23L 1/236, A23L 1/22, publ. 20.04.2005).

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

DISCLOSURE of INVENTIONS

Object of the invention is the creation of an immunostimulating drug-based reveal aimed at treatment and prevention of immune States caused as pathological processes, and unfavorable environmental conditions. Contributing to the increase in General and the specific resistance is organisma, developed the drug is cheap and highly effective for correction of immune status of animals, thereby reducing the risk of death from diseases of different 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 organism.

The technical result is achieved by using a method of obtaining immunostimulating drug for animals, comprising preparing the reveal, with stevioside dissolved in 0.9% saline solution, the resulting solution was added to pre-powdered lecithin, and then the resulting mixture was gradually heated to contribute 35-40°With olive oil and mix, then make filtering the resulting solution and autoclave at a temperature of 115-120°C for 15-20 minutes, then obtained a sterile preparation is poured into bottles and corked.

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

Stelios is d 10-30
A physiological solution of 0.9%9-11
Lecithin1-3
Olive oilrest

For the preparation of immunostimulating drug for animals as the main active ingredient was chosen stevioside. Stevioside - diterpenoid glucoside having cardiotonic, hypoglycemic and immunostimulating action on the animal organism. (Sehar I, and all. "Up Immune regulatory response of a non-CR natural sweetener, stevioside" Chem Biol Interact. 2008 May 28; 173 (2): 115-21. Epub 2008 Jan 31). Saline solution necessary to dissolve and reveal the emulsion. Lecithin is an affordable and effective emulsifier. Olive oil is the medium for the preparation of immunostimulant and causes prolonged effect of the drug on the animal organism.

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

10, 20 and 30 (wt.%) the reveal is dissolved in 9-11 (wt.%) a 0.9% saline solution, the resulting solution was added to pre-pulverized in a mortar lecithin 1-3, then the resulting mixture was gradually make up to 100% heated to 35-40°With olive oil and mix, then make filtrowanie the resulting solution through a paper filter and autoclave at a temperature of 115-120°C for 15-20 minutes, received sterile drug poured into vials and sealed, the ratio of components, wt.% is:

Stevioside10-30
A physiological solution of 0.9%9-11
Lecithin1-3
Olive oilrest

The IMPLEMENTATION of the INVENTION

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

Example 1. 5 (wt.%) the reveal is dissolved in 9 parts of 0.9% saline solution, the resulting solution was added to pre-pulverized in a mortar lecithin, and then the resulting mixture was gradually heated to contribute 20°With olive oil and mix, then make filtering the resulting solution and autoclave at a temperature of 80°C for 10 minutes, the resulting preparation is poured into vials and sealed, the ratio of components, wt.% is:

td align="right"> 9
Stevioside5
A physiological solution of 0.9%
Lecithin1
Olive oilrest

With the introduction of the drug to animals no reactions from the immune system is not mentioned, oil, warmed to 20°C, poorly mixed with the other components autoclaving at a temperature of 80°C for 10 minutes is not enough, as in the injection site appeared obvious signs of inflammation.

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

Stevioside7
A physiological solution of 0.9%9
Lecithin1
Olive oilrest

With the introduction of the drug to animals no immunostimulating effect was not observed, the oil is heated to 25°C, very slowly mixed with the other components autoclaving at a temperature of 90 the 15 minutes is not enough as the injection site occurred hyperemia.

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

Stevioside10
A physiological solution of 0.9%10
Lecithin2
Olive oilrest

There is a pronounced effect of the drug on the immune system, increases the number of leukocytes, is not observed no negative effect on the organism of experimental animals. The drug due to the small concentrations reveal convenient to use small animals (cats, dogs, rabbits and other).

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

Stevioside
20
A physiological solution of 0.9%10
Lecithin2
Olive oilrest

This concentration of reveal and method of preparation of the drug is the most effective, as it is observed the most pronounced effect of the drug on the immune system, and have not observed any negative effect on the organism of experimental animals. This concentration of reveal in the product easy to use animals of medium size (sheep, goats, pigs and others).

Example 5. The product is prepared analogously to example No. 4, but use almond oil, the ratio of components, wt.% is:

Stevioside20
A physiological solution of 0.9%10
Lecithin2
Almond oilrest

Noted the high cost of the drug due to the scarcity and dearness of almond oil.

Example 6. The drug is prepared similarly, when the ERU No. 4, but dissolving 30 (wt.%) the reveal in 11 ml of 0.9% saline solution, olive oil heated to 40°C, autoclave at a temperature of 120°C for 20 minutes, the ratio of components, wt.% is:

Stevioside30
A physiological solution of 0.9%11
Lecithin3
Olive oilrest

The increase in the number of T - and b-lymphocytes in the background of the harmlessness of the drug on the animal organism, indicates the efficiency and the concentration of the reveal. This method of preparation of a drug is effective, but slightly long. Such a concentration reveal it is advisable to use large animals (cattle, horses and others).

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

Stevioside 40
A physiological solution of 0.9%15
Lecithin4
Olive oilrest

This concentration of reveal has caused the violation of the biochemical processes and showed undesirable effect on the immune system of animals. The increase in the share of saline increased the size of the dispersion phase in the emulsion. The decay rate of the emulsion is not decreased by increasing the proportion of emulsifier. Increasing the temperature, time of autoclaving and the proportion of emulsifier increased the cost of production.

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

Stevioside50
A physiological solution of 0.9%25
Lecithin5
Olive oilrest

Histological examination of the organs of experimental animals after application of such concentrations reveal revealed degenerative changes in the liver. Obvious overuse of emulsifier and saline solution, emulsion contains large droplets of the dispersion phase and quickly breaks down into fractions. Time, temperature autoclaving and wasted emulsifier increases the cost of production.

The TEST RESULTS IMMUNOSTIMULATING DRUG FOR ANIMALS

Developed the drug was tested on sheep 9-10 months of age Stavropol breed. Drug created on the basis of biologically active substances (stevioside), was introduced clinically healthy sheep intramuscularly at a dose of 2 ml per 50 kg of body weight three times every three days. Blood samples from sheep were taken from the jugular vein on day 10 after the last injection of the drug. Was determined in the blood of the amount of hemoglobin, erythrocytes and leukocytes, serum level of total protein, albumin, α - β - and γ-globulins, as well as indices of immunological reactivity (the content of T - and b-lymphocytes, lysozime, phagocytic and bactericidal activity). The results of the test drug on the basis of the reveal are presented in tables 1, 2, 3 and 4.

Tab the Itza 1
Hematological blood parameters of sheep (M±m)
The group of animalsHemoglobin, g/lRed blood cells, ×1012/lThe leukocyte count, ×109/l
Control107,3±0,68,6±0,26,4±0,3
Experienced112,6±1,78,7±0,18,5±0,3

Based on the data presented in table 1, revealed an increase in the number of cells at 33% (8,5±0,3 ×109/l) in the experimental animals, the amount of hemoglobin and red blood cells has not been significantly changed.

Table 2
Biochemical indices of the blood of sheep (M±m)
The group of animalsTotal protein, g/lAlbumin, g/lThe globulin, g/l
α, g/lβ, g/lγ, g/l
Control/td> 68,2±1,216,9±4,913,7±1,812,7±2,0525,1±5,4
Experienced63,3±1,712,3±2,0313,1±2,98,7±1,629±4,03

From table 2 it is seen that an immunostimulating drug for animals did not have a significant impact on these biochemical parameters like total protein content and α-globulin. Was observed in 13% (29±4,03 g/l) increase relative to the control group, γ-globulin, a 30% reduction in the number of β-globulin (8,7±1.6 g/l) and 27% reduction in albumin (12,3±2,03 g/l).

Table 3
Immunological blood parameters of sheep (M±m)
The group of animalsBASK, %FASK, %FOREPLAY, %T-lim., %B-lim., %
Control7,4±0,434,6±8,917,9±0,560,2±2,4 15,8±0,6
Experienced22,8±1,140±5,520,3±1,279,6±3,636,6±3,7

Analyzing immunological parameters, presented in table 3, it should be noted the increase in bactericidal (BASK) and lysozyme (FOREPLAY) activity of blood serum of animals of the experimental group, 67% (22,8±1,1%) and 11% (20,3±1,2%), respectively. Phagocytic activity of neutrophils (FASK) relative to the control group increased by 13% (40±5.5 per cent). The relative amount 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
The productivity and safety of animals (M±m)
The group of animalsLive weight, kgIncidence, %Mortality, %Obtaining a yield goal. per 10 females
control46±2,513±0,22,8±0,711±0,7
the experience of the traveler 53±1,68±0,11,3±0,413±0,3

Analyzing the data presented in table 4, it should be noted that in the experimental group animals by 38% decreased morbidity and 53% of the deaths of animals. After application of the drug based on the reveal on a 13% increase in live weight of animals and a 15% increase in the birth rate.

Based on the above research results, we can recommend an immunostimulating drug for animals for veterinary use, as immunostimulatory agent, which is able to act on specific and non-specific defense system of the organism.

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

- improve the viability and productivity of animals due to the correction of the immunological reactivity of the organism;

- low cost and environmental friendliness of the product;

- has a prolonged effect on the animal organism;

- the possibility of injecting;

different concentrations of the active substance.

1. The method of obtaining immunostimulating drug for animals from the reveal, with stevioside dissolved in 0.9%physiological solution from the first solution is added to pre-powdered lecithin, then, the resulting mixture was gradually heated to contribute 35-40°With olive oil and mix, then make filtering the resulting solution and autoclave at a temperature of 115-120°C for 15-20 min, then obtained a sterile preparation is poured into bottles and corked.

2. Immunostimulating drug for animals, including stevioside, characterized in that it further comprises a 0.9%saline solution, olive oil, lecithin when the ratio of components, wt.%:

Stevioside10-30
A physiological solution of 0.9%9-11
Lecithin1-3
Olive oilrest



 

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