RU2404761C1 - Method for production of complex preparation for prophylactics and treatment of metabolic derangements, microelementoses, improvement of animal organism resistance - Google Patents

Abstract

FIELD: medicine, veterinary science.

SUBSTANCE: invention is related to veterinary science. Method for production of complex preparation for prophylactics and treatment of metabolic derangements, microelementoses, improvement of animal organism resistance includes mixing of water-soluble salts of metals - stimulators of iron, zinc, copper, cobalt with sodium nucleinate as immunostimulant, succinic acid and methionine at the following ratio of components, in the following amounts, g/l: Succinic acid 10.0; Iron sulfate 2.0; Copper sulfate 0.1; Cobalt sulfate 1.0; Zinc sulfate 0.1; Sodium nucleinate 10.0; Methionine 10.0; Water for injections up to 1000 ml and pH of 6.8-7.0.

EFFECT: invention provides for production of efficient preparation for parenteral introduction for prophylactics and treatment of metabolic derangements, microelementoses, improvement of animal organism resistance.

4 tbl, 3 ex

Description

The invention relates to veterinary medicine, in particular to methods for complex preparations for the prevention and treatment of metabolic pathologies and increase the resistance of animals.

Known complex anti-anemic and growth-promoting drug for animals "Gemovit-plus" (Kozlov Yu.M. RF Patent PU 203657 from 06/04/2002). The Hemovit-plus preparation contains a disodium or dipotassium salt of ethylenediamine - N, N ¹ -disuccinic acid, water-soluble salts of iron, manganese, copper, cobalt, zinc and selenium.

Known comprehensive anti-anemic and growth-promoting drug "Hemovit-meyan" (Babich V.A. et al. RF Patent PU 225 2020 dated 05.02.2005). The Hemovit-Meyan preparation includes a chelate complex of iron, manganese, zinc, copper, cobalt, selenium and iodine with an organic ligand. As a ligand, it contains the trisodium salt of methionine succinic acid.

An important distinguishing feature of these drugs is that biogenic metals are associated with succinic acid in a complex compound of the chelate type. At the same time, succinic acid serves not only as a carrier of biogenic metals, but also is a metabolite and a stimulator of metabolic processes. These drugs are intended for oral administration. However, the oral method of administration, despite its simplicity, has a major drawback - a significant part of the salts of trace elements (up to 70%) does not have time to be absorbed by the body and excreted with excrement. In this connection, to achieve a therapeutic effect, a long course of use of the drug is required, which cannot be considered the optimal solution to the problem.

A known method for producing a complex metal-globulin preparation (Klenina N.V. et al. A method for producing a complex preparation of immunoglobulins with metals. A.S. SU 1352696 USSR of 06.26.1985). The combination of immunoglobulins with metals ensures their good bioavailability with the parenteral method of administration, and therefore the drug has a pronounced immunostimulating effect. However, the drug is not effective enough to normalize metabolic processes in case of impaired liver function. In addition, in the manufacture of immunoglobulin inevitably imposed increased health requirements for animals used as donors in the collection of blood serum. At the same time, to ensure the sterility of the drug, the use of a more reliable and simple method of sterilization by autoclaving is excluded.

The objective of the present invention is to provide a comprehensive preparation effective for the prevention and treatment of metabolic disorders, microelementoses, enhance the hepatoprotective function of the liver, increase the resistance of the animal organism.

The problem is achieved by the inclusion in the complex preparation of water-soluble salts of metal stimulants: iron, copper, zinc, cobalt with the activator and carrier of these salts with succinic acid.

As an immunostimulant, sodium nucleinate is included in the composition of the drug.

Sodium Nucleinate is a synthetic immunostimulant. It is used for the prevention and treatment of infectious diseases occurring against the background of immunodeficiency states. Thermostable.

To enhance hepatoprotective activity, methionine is included in the composition of the drug.

Methionine is an essential amino acid and is a “critical” amino acid for newborn animals, since their digestive system cannot synthesize it from nitrogen-containing feed substances. With a lack of methionine, anemia, fatty and toxic liver dystrophy are often observed. In the body, methionine activates the action of a number of enzymes, hormones, vitamin B ₁₂ , folic and ascorbic acids. Many processes of neutralization of toxins in the liver also occur with the participation of methionine.

The proposed drug allows you to get both metabolic, hepatoprotective, immunostimulating effects. The components included in its composition are affordable and standard. Their most optimal ratio is, g / l:

Succinic acid - 10.0

Ferrous Sulfate - 2.0

Copper Sulfate - 0.1

Zinc Sulfate - 0.1

Cobalt Sulfate - 1.0

Methionine - 10.0

Sodium Nucleinate - 10.0

Water for injection - up to 1000 ml

This ratio of succinic acid, metal salts, methionine and sodium nucleinate was found experimentally and provides a synergistic effect with the parenteral (injection) method of administration. Autoclaving sterilization does not change the physicochemical properties and biological activity of the drug.

The technological process of obtaining a complex preparation is illustrated by an example.

Example

To obtain 1 liter of the drug, 800-850 ml of water for injection is poured into the flask. 10.0 g of succinic acid are added to it and heated to 50-60 ° C. Samples containing 2.0 g of iron sulfate, 0.1 g of copper sulfate, 0.1 g of zinc sulfate, 1.0 cobalt sulfate, 10.0 g of methionine, 10 g of sodium nucleate are successively added to the heated solution with stirring. The final volume is adjusted to 1000 ml by adding water for injection.

The resulting solution is poured into dark glass bottles with a capacity of 50 ml and sterilized for 25 minutes.

The composition obtained in the above example was sterile. When the samples of the composition were inoculated on meat-peptone broth (MPB) and MPB under vaseline oil in test tubes and on meat-peptone agar (MPA) in Petri dishes, followed by exposure to an incubator at 37 ° C for 48 hours, microflora was not grown.

When checking the acute and subchronic toxicity of the drug in white mice, it was found that LD ₅₀ was 22500 mg / kg body weight, which allows it to be classified as a low-hazard drug.

The resulting preparation, which we called metallosuccinate plus, when tested on animals, induced a number of vital biological effects, which are illustrated by the corresponding examples.

Example 1

A study of the hematological and growth-stimulating activity of metallosuccinate plus and a drug of the same composition, but without methionine, was carried out in comparison with ferroglukin in newborn piglets. At 4 days of age, piglets of the first experimental group (n = 9) were intramuscularly injected with metallosuccinate plus in a volume of 1.5 ml, piglets of the second experimental group (n = 9) were injected with the basic version of metallosuccinate (without methionine) in the same volume, and piglets of the control groups (n = 7) in the same dose was administered ferroglukin.

At 8 days of age, drug injections were repeated, increasing their dose to 2.0 ml. Blood was taken for hematological examination on the day of the first injection of drugs and 7 days after repeated administration.

The results of the studies are presented in table 1.

Table 1 Blood counts of piglets before and after drug administration Indicators Experimental group No. 1 Experimental group No. 2 Control group before introduction / after before introduction / after before introduction / after Red blood cells, mln / ml

Hemoglobin, g%

Bactericidal activity of blood serum (BASK),%

Phagocytic activity of leukocytes (FAL),%

The results of hematological studies indicate that metallosuccinate plus had a more pronounced stimulating effect on the hematopoiesis system than metallosuccinate without methionine and was superior in efficiency to the iron-containing preparation ferroglukin. In the blood of piglets of the first experimental group, indicators of the content of red blood cells and hemoglobin were significantly higher relative to the level of their peers from the second experimental and control groups. A reliably high level of indicators of natural resistance of BASK and FAL in piglets of the experimental groups indicates a pronounced stimulation of factors of nonspecific resistance of the immune system. It should be noted that in piglets of the first experimental group for 15 days (daily observation period) was not a single case of diarrhea. In 3 cases out of 8 piglets of the second experimental group, a slight course of diarrhea was recorded, while all piglets in the control group constantly showed signs of dyspepsia, which could not but affect their growth activity.

At 30 days of age, the average weight of piglets in the first experimental group was 4.8 kg, in the second experimental group - 4.5 kg versus 3.9 kg in animals in the control group.

The data obtained convincingly indicate that metallosuccinate in combination with methionine has a well-defined stimulating effect both on the hematopoiesis system and on the intensity of metabolic processes.

Example 2. The study of the effect of metallosuccinate plus on the biochemical status of cows with hepatosis

During the formation of the experimental groups, a targeted selection of animals that showed signs of digestion, an increase in the posterior border of the liver, resorption of the last caudal vertebrae, and bile pigments and urobilin in the urine was performed. All of the above signs gave the basis to consider cows with hepatosis.

To establish the degree of imbalance in metabolic processes, biochemical blood tests were performed. Based on the results of biochemical studies, the final selection of experimental animals was carried out, which had similar metabolic rates, indicating a low functional state of the liver.

Experimental cows of the first group (n = 9) were injected intramuscularly with metallosuccinate plus three times with an interval of 24 hours in a volume of 10.0 ml; cows of the second experimental group (n = 7) were injected with cows of the second experimental group without methionine. The cows of the control group (n = 7) were injected with saline in the same way. Control of changes in metabolic processes was carried out on the basis of biochemical studies of blood serum on the 7th and 14th day after the injection of drugs. The results of the studies are shown in table 2.

table 2 The effect of metallosuccinate plus on metabolic processes in cows with hepatosis Indicators Preparations Metallosuccinate Plus Metallosuccinate (without methionine) Saline Protein, g / l before introduction 10.46 ± 1.26 10.58 ± 1.13 10.42 ± 0.73 after 7 days 8.12 ± 0.73 8.37 ± 0.86 10.50 ± 0.25 after 14 days 8.46 ± 0.25 8.50 ± 0.92 10.74 ± 0.38 Reserve alkalinity, mg% before introduction 26.81 ± 3.21 26.91 ± 1.46 25.92 ± 3.46 after 7 days 56.8 ± 3.46 54.23 ± 3.12 26.2 ± 2.98 after 14 days 55.2 ± 2.74 53.81 ± 2.32 27.4 ± 2.12 Ketone bodies, mg% before introduction 20.26 ± 2.34 20.34 ± 2.76 29.64 ± 2.36 after 7 days 8.74 ± 1.12 12.64 ± 1.94 20.12 ± 1.94 after 14 days 6.92 ± 1.25 8.12 ± 1.12 20.17 ± 2.12 Calcium, mg% before introduction 7.24 ± 0.83 7.32 ± 0.54 7.24 ± 0.56 after 7 days 9.72 ± 0.48 8.96 ± 0.38 7.26 ± 0.34 after 14 days 10.46 ± 0.12 9.80 ± 0.14 7.28 ± 0.25 Phosphorus, mg% before introduction 6.98 ± 0.24 7.97 ± 0.11 6.84 ± 0.36 after 7 days 5.39 ± 0.71 5.46 ± 0.28 6.79 ± 0.17 after 14 days 5.25 ± 0.36 5.12 ± 0.22 6.58 ± 0.24 Iron, mg% before introduction 3.27 ± 0.18 3.41 ± 0.16 3.39 ± 0.25 after 7 days 5.35 ± 1.21 5.25 ± 1.12 3.42 ± 0.17 after 14 days 4.98 ± 0.83 4.78 ± 0.42 3.43 ± 0.25 Copper, mg% before introduction 227.81 ± 5.39 236.02 ± 2.53 236.61 ± 2.53 after 7 days 320.42 ± 3.46 232.14 ± 3.25 232.82 ± 3.25 after 14 days 396.71 ± 3.15 278.41 ± 4.36 214.18 ± 4.18 Zinc, mg% before introduction 9.23 ± 0.32 9.21 ± 0.26 9.16 ± 0.18 after 7 days 14.72 ± 1.21 14.19 ± 1.46 8.91 ± 0.46 after 14 days 14.61 ± 1.35 13.21 ± 0.94 8.83 ± 0.27 Cobalt, mg% before introduction 7.23 ± 0.28 7.21 ± 0.14 7.25 ± 0.15 after 7 days 9.65 ± 0.76 9.13 ± 0.82 7.19 ± 0.16 after 14 days 9.43 ± 0.65 8.92 ± 0.38 7.16 ± 0.12

During clinical observation, it was found that one day after the first injection of metallosuccinate plus and metallosuccinate without methionine, cuds and scar reduction were activated in cows. On the 5th day, their appetite increased significantly, chewing gum and scar reduction completely recovered, and a pronounced tendency to increase milk productivity was also noted.

During biochemical blood tests on days 7 and 14 after the start of the experiment, the level of ketone bodies decreased by 2-3 times in all the cows of the first experimental group, which indicated a marked improvement in the functional activity of the liver. In almost all animals of the first experimental group, there was a complete normalization of the content of protein, calcium, phosphorus, reserve alkalinity, which indicated a high metabolic activity of the test drug. Less pronounced, especially with regard to the level of ketone bodies in the blood, changes were observed in cows that were given metallosuccinate without methionine. In cows of the control group, no positive changes were noted.

A pronounced increase in blood levels of iron, copper, zinc, cobalt in animals of the experimental groups indicated an increase in their digestibility by the body. In the control group, the content of trace elements did not significantly change.

Activation of metabolic processes, a marked improvement in the functional activity of the liver, elimination of the acidotic state positively affected the milk production of cows. In the period from 10 to 30 days, the daily milk yield from each cow in the first experimental group increased by 3.5 liters, in cows of the second experimental group - by 2 liters. In the cows of the control group, milk productivity during this period remained almost at the same level. At the same time, the cows of the experimental groups looked better, their hairline was even, while in the control animals it was tousled and dull.

Example 3. The effect of metallosuccinate plus on the immunological reactivity and nonspecific resistance of calves with signs of malnutrition

The experiment was carried out on a month old calves with clinical signs of malnutrition. The first experimental group of calves (n = 7) was injected twice, intramuscularly, with an interval of 24 hours, in a volume of 3.0 ml, metallosuccinate plus injections were given, the calves of the second experimental group (n = 7) were injected with metallosuccinate without methionine according to a similar scheme. The calves of the control group (n = 5) were injected with saline in the same way. Control immunological studies were carried out on 5-7 days. The research results are presented in tables 3 and 4.

Table 3 Dynamics of immunological parameters of calves hypotrophic under the influence of metallosuccinate plus Indicators Preparations Metallosuccinate Plus Metallosuccinate (without methionine) Saline White blood cells, g / l

Lymphocytes,%

T-lymphocytes,%

B-lymphocytes,%

IgM

IgG

Note. Numerator - initial data, denominator - after 7 days.

Table 4 Dynamics of factors of nonspecific resistance of calves hypotrophic under the influence of metallosuccinate plus Indicators Preparations Metallosuccinate Plus Metallosuccinate (without methionine) Saline BASK,%

Lysozyme, mg / ml

FAL,%

Note. Numerator - initial data, denominator - after 7 days.

Based on the analysis of the initial data of table 3, the calves had all the signs of an immunodeficiency state. In addition, a very weak activity of factors of nonspecific resistance was revealed (see Table 4).

The results of the use of metallosuccinate plus revealed a well-defined stimulating effect on the cellular and humoral systems of immunity and nonspecific resistance, which can have a positive value both in the complex accompanying therapy of infectious diseases and in the induction of anti-infection protection.

Claims (1)

A method of obtaining a comprehensive preparation for the prevention and treatment of metabolic disorders, microelementoses, increasing the resistance of the animal organism, including mixing water-soluble metal salts - stimulants of iron, zinc, copper, cobalt with an immunostimulant, characterized in that sodium nucleate is introduced as an immunostimulant and amber is additionally administered acid and methionine in the following ratio of components, calculated in g / l and pH 6.8-7.0:
succinic acid 10.0 Iron sulfate 2.0 Copper sulphate 0.1 Cobalt sulfate 1,0 Zinc sulphate 0.1 Sodium Nucleinate 10.0 Methionine 10.0 Water for injections Up to 1000 ml