RU2155052C1 - Complex antiparasitic preparation without immunosuppression effect - Google Patents

Abstract

FIELD: biology, pharmacology, veterinary science. SUBSTANCE: invention proposes the complex preparation that has avermectin complex as avermectin N from Streptomyces avermitilis VKPM S-1440 mycelium as an antiparasitic agent with increased content of component of the group B (67.8-77.4%). Also, other avermectin complexes (aversectin C, ivermectin or other aver-mectin-base active substances) can be used as drugs. Ribotane is used as an immunostimulating and antiinflammatory agent that enhances body-host resistance and prevents topical response. Ethyl alcohol and benzyl alcohol are used as solvent. Polyvinyl-pyrrolidone and polyethylene glycol-400 are used as disintoxicating, chelating agent that prolongs curative effect of the preparation. Also, distilled water is used and all components are taken in the definite ratio. Preparation is administrated by subcutaneous route and after one-two dose it provides 100% curative antiparasitic effectiveness being without suppression of immune system and topical response in animals. Invention can be used for prophylaxis and treatment of animals with diseases caused by mites, insects, nematodes of different species. EFFECT: enhanced effectiveness of preparation. 3 cl, 1 tbl, 7 ex

Description

 The invention relates to veterinary medicine, pharmacology, biology and is intended for the treatment and prevention of parasitic diseases of animals caused by different types of ticks, insects, nematodes.

 Known means for the treatment of parasitic diseases of animals, including avermectins and carriers (1, 2, 3, 4, 5, 6, 7, 8).

 The closest in technical essence to the claimed invention is the preparation of an avermectin complex for the prevention and complex treatment of diseases caused by endo- and ectoparasites of animals, containing the active principle from the mycelium Streptomyces avermitilis VNIISKHM-54 and a pharmacologically acceptable carrier (2,3). In the injectable dosage form, the preparation contains 18-20 g / l of the avermectin complex in ethanol, including 36.75 ± 1.35% of the components of group B (B1 + B2), and the rest is a pharmacologically acceptable carrier. The disadvantage of foreign and domestic injection forms based on avermectins is the suppression of the animal immune system for 2-3 weeks (9), increased individual sensitivity of some animals, accompanied by local swelling at the injection site, which takes place after 3-7 days (10).

 The aim of the present invention is the creation of an injectable drug with not only high antiparasitic activity due to the higher content of group B avermectins, but also an immunostimulating effect and the absence of local irritant effect.

 The solution to this problem is achieved by introducing an avertin N obtained from Streptomyces avermitilis VKPM-S-1140 as an antiparasitic agent of the avermectin complex, but the avermectin complexes used in other medicines, referred to as either aversectin C or abamectin, can be equally used , or ivermectin, or other active substances based on avermectin. As an immunostimulating and anti-inflammatory agent that increases the resistance of the host organism and prevents a local reaction, ribotan is used (other immunostimulants can be used); as a solvent - ethyl alcohol, benzyl alcohol; as a detoxifying complexing agent, prolonging the action of the drug, polyvinylpyrrolidone (PVM), polyethylene glycol 400; distilled water is used as a filler in the following ratios of components, wt.% (table. 1).

The avermectin complex is produced by soil microorganisms Streptomyces avermitilis, belonging to the family of avermectins from the group of macrocyclic lactones. It consists of 4 main (B _1a , B _b , A _1a , A _1b ) and 4 minor (B _2a , B _2b , A _2a , A _2b ) components that are closely interconnected. The components of group B have a pronounced antiparasitic effect. The advantage of the strain of Streptomyces avermitilis VKPM-S-1140 is that it produces 67.8 - 77.4% of the components of group B (15), in contrast to the strain of Streptomyces avermitilis VNIISKHM-54 - 55, 63 - 58.65% (2.3).

 For the production of an antiparasitic drug that does not have immunosuppression and local irritating effects, but exhibits high parasitocidal activity, the avermectin complex from the Streptomyces avermitilis strain VKPM-S-1140 (the active substance is Avertin N) was used. Avermectin complexes can be used equally: either aversectin C, or abamectin, or ivermectin, or others. All avermectin complexes in a very low therapeutic dose of 0.2 mg / kg body weight have a wide spectrum of antiparasitic activity (4).

 Ribotan is a complex immunocorrector consisting of low molecular weight peptides (MM 0.5 - 1.0 kDa) and low molecular weight RNA fragments. The immunostimulating effect of ribotan is aimed at increasing the functional activity of macrophages, subpopulations of T and B lymphocytes, as well as the synthesis of interferon and lymphokines. Indications for the use of ribotan are parasitic, viral and bacterial infections of animals.

Ethyl alcohol is a solvent. Miscible in all ratios with water, ether, chloroform, glycerin; boils at 78 ^° C (13.14).

Benzyl alcohol is a solvent. It is difficult to dissolve in water, boiling point 128-132 ^o C (14).

 Polyethylene glycol-400 is a good solvent and a good complexing agent, which is often used as a component of the dosage form (12, 13).

 Polyvinylpyrrolidone (PVP) - low molecular weight (12600 + 2700), medical is a powder soluble in water, 95% alcohol, sparingly soluble in carbohydrates and practically insoluble in ether. PVP is used for the preparation of the blood-substitute drug "Hemodez" with a detoxifying effect. PVP forms complexes with a number of pharmaceuticals and helps to prolong their action (13).

 Distilled water - get it by distillation of drinking water using special distillation apparatuses (13).

 From the sources of information available to the applicant, the indicated set of essential features is not known, which makes it possible to obtain the indicated technical result, therefore, the claimed invention meets the patentability condition “novelty”.

 The applicant does not know means of a similar purpose, in which the distinguishing features of the claimed invention would be disclosed with the use of it in any combination of features of the specified technical result. Therefore, the claimed invention meets the condition of patentability "inventive step".

 Evidence of industrial applicability of the claimed drug is given in the following examples.

Example 1. In a 5-liter container, 1.0 kg of PAP (intermediate of antiparasitic preparations from Streptomyces avermitilis) and 1.8 L of 75% ethanol are placed. The mixture is stirred for 1 hour, then filtered, 1.5 kg of PEG added to the filtrate, stirred for 10 minutes and placed in a refrigerator for 24 hours at a temperature of 30-40 ^o C. Then the mixture is placed in a container with a lower descent and kept at room temperature for 48 hours. During this time, the mixture is stratified into 2 layers. For further work, the bottom layer of the mixture is taken, to which 0.5 l of ribotan, 0.2 kg of PVP are added, distilled water to a volume of 5 l and stirred for 20 minutes, then the drug is poured into a container.

 Example 2. In a 5-liter container, 1.6 kg of PAP, 1.45 L of 75% ethyl alcohol, 0.25 L of 75% benzyl alcohol are placed. The mixture is stirred for 1.2 hours, filtered, 1.0 kg of PEG are added, followed by Example No. 1, then 0.3 kg of PVP, 0.4 L of ribotan and distilled water are added. The result is a preparation of the following composition, wt. %: avermectin complex 1.0; ethyl alcohol 40.0; benzyl alcohol 4.0; polyethylene glycol 40.0; polyvinylpyrrolidone 2.0; distilled water - the rest.

 Example 3

 The medicinal composition is prepared according to example No. 1 with the following composition of components: 0.5 kg of PAP, 2.0 l of ethyl alcohol, 0.5 l of benzyl alcohol, 0.2 kg of PEG, 0.8 kg of PVP and 1.0 l of ribotan.

 The proposed complex preparation is tested on different types of animals, both intact and affected by ecto- and endoparasites.

 Example 4. Immunomodulating effect of the complex drug on intact mice.

 Of 40 white outbred mice weighing 20 g, 5 equal groups were formed. The mice of the first group were injected with avertin at a dose of 0.18 ml / goal. (the drug was diluted 300 times with polyethylene glycol), the second - ribotan at a dose of 10 μg / goal., the third - a complex drug at a dose of 0.1 ml / goal., the fifth group was given a placebo.

 The primary immune response to the thymus-dependent antigen was determined at 7 and 10 days after drug administration. The antibody titer in animals of the first group was 4.00 ± 0.40, in the second - 4.83 ± 0.19, in the third - 4.68 ± 0.17, in the fourth - 4.40 ± 0.12, against 3 , 62 ± 0.40 in the control. Therefore, the action index (ID) was 0.38 in the first group, which confirms the suppressive effect of the drug, in the second - 1.21, in the third - 1.08, in the fourth - 0.82.

 Thus, a complex preparation and ribotan at a dose of 10 μg / mouse have a pronounced immunostimulating property, and a complex preparation at a dose of 5 μg for ribotan does not have such properties.

 Example 5. The effect of the complex drug on the immune status of piglets.

 Of the 20 piglets, according to laboratory studies infected with hematopinuses and roundworms, 2 groups were formed. The animals of the first group (12 animals) were injected subcutaneously with a single complex preparation in order to treat invasive diseases and increase the immune status, and 8 animals of the second group served as a control.

An immunological examination found that in piglets of the first group the number of T-immunocytes ranged from 44.2 ± 4.2 to 51.2 ± 2.8 within 20 days after administration of the drug, the stimulation index (the ratio of T-helpers to T-suppressors ) ranged from 3.2 to 4.4, the number of B-lymphocytes was in the range of 26.8 ± 4.2 to 29.4 ± 3.2 also on the 5th, 10th and 20th days of observation. The titers of natural antibodies were log ₂ 4.0 ± 0.1, log ₂ 4.4 ± 0.2 and log ₂ 4.6 ± 0.4, respectively, at 5, 10 and 20 days after administration of the drug.

 The effectiveness of the complex drug against hematopinuses and roundworms was 100%.

In the control group, the number of T-lymphocytes was 22.8 ± 4.2 - 24.6 ± 3.4 - 23.8 ± 2.4%, respectively, on the 5th, 10th and 20th days of the study, IP was 0.6 - 0, eight. The number of B-lymphocytes was 14.4 ± 2.2 - 14.0 ± 0.1 and 12.4 ± 1.1%, and the titers of natural antibodies from 2.0 ± 0.2 to 2.4 ± 0.1 log ₂ .

 Animals of the control group for the entire observation period isolated Ascaris eggs and there were many hematopinuses on the skin.

 Thus, the complex preparation has a stimulating effect on T- and B-systems of immunity, on a subpopulation of T-lymphocytes, causes an increase in the titer of natural antibodies and has a 100% effect against parasitic diseases of pigs.

 Example 6. Anthelmintic and immunostimulating activity of a complex preparation. Of the 20 lambs at the age of three months, spontaneously invaded 100% with strongilates of the gastrointestinal tract and strongyloids, 2 groups were formed. The animals of the first group were injected subcutaneously once a complex preparation made according to example No. 1 at a dose of 1 ml / 50 kg of body weight, and the animals of the second served as a control and the drug was not received. When assessing the immune status, it was found that the number of T-lymphocytes in the experimental lambs was from 37.8 ± 5.8 to 44.4 ± 4.2%, IP from 2.6 to 4.2, the number of B-lymphocytes from 21, 4 to 25.2%. Bactericidal activity and titers of natural antibodies were also increased. In the study of feces by the methods of Fulleborn and Berman, eggs of strongilates and strongyloids were not found in animals of this group. All control animals 100% isolated eggs of different species.

 The results of the experiment are presented in table. 2.

Thus, the introduction of a complex drug prevents nematodoses, increases the immune status and general resistance of animals
Example 7. Therapeutic and immunostimulating efficacy of a complex preparation for sheep psoroptosis.

 Of the 20 sheep with damaged skin surface areas with Psoroptes ovis scabies, 4 equal groups were formed. The sheep of the first group were injected subcutaneously once with the preparation prepared according to example 1, the second - according to example 2; the third is a drug containing 1.0% DV and 10 mg of ribotan; the fourth is a drug containing 1.0% DV, but without ribotan. After 8 days, the injection of the drug was repeated.

 In the sheep of the first three groups, after the first injection, by the fifth day, a marked improvement in the clinical condition appeared (the itching stopped), in the sheep of the fourth group - by the 10th day. After the second injection in animals of the first three groups, full recovery occurred after 40 days, the fourth - after 50 days.

 The first three drugs included an immunostimulant, which in combination with avermectins had its positive effect.

 Thus, the antiparasitic composition of the proposed composition, which includes an avermectin complex and an immunostimulant, has not only a wide spectrum of high antiparasitic action, but also prevents immunosuppression in comparison with just avermectins.

 The combination of components used in the proposed ratio provides high antiparasitic anti-inflammatory effectiveness and increases the resistance of the host organism.

Literature
1. William C. Campbell. lvermectin and abamectin. / New York, Berlin, Heidelberg, London, Paris, Tokyo, 1989, p. 1-25, 114-124.

 2. RF patent N 2054483, 1993.

 3. RF patent N 2054848, 1996.

 4. Berezkina S. V., Golovkina L. P., Drinyaev V. A., Yurkiv V. A., Volkova G. N. Natural avermectins for the treatment of ecto- and endoparasites of animals. Mater. conf. "Systematics, taxonomy and fauna of parasites", M., 1996. - S. 15-16.

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Claims (3)

 1. A complex antiparasitic drug that does not have immunosuppression, including an avermectin complex and excipients, characterized in that it additionally contains an immunostimulant ribotan in the following ratio, wt.%: Avermectin complex 0.5 - 2.0; ribotan 5.0 - 15.0; excipients - the rest.  2. The complex antiparasitic preparation according to claim 1, characterized in that, as an avermectin complex, it contains either Avertin N, or Aversectin C, or Ivermectin, or Abamectin, or others.  3. The complex antiparasitic preparation according to claim 1 is characterized in that it comprises an avermectin complex from Streptomyces avermitilis mycelium VKPM S - 1440 with an increased content of group B.

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