RU2681214C1 - Method of obtaining agent for the treatment of solid-hoofed animals with parasitosis - Google Patents

Abstract

FIELD: veterinary medicine.SUBSTANCE: group of inventions relates to veterinary medicine, in particular, to a method for producing an agent for treating solid-hoofed animals with parasitosis, as well as to the agent for treating solid-hoofed animals and the method of treating solid-hoofed animals with nematodosis and botts of the digestive tract. In the laboratory, the preparation is obtained in stage 3, in the first stage, sugar syrup is prepared in a separate container by heating water to +50 °C, with stirring with the addition of sucrose and sodium alginate, in the second stage, in another container at a temperature of +50 °C prepare a solution of an intermolecular complex of ivermectin based on polymers, PEG-400, propylene glycol, PVP-17, in the third stage, a solution of ivermectin is added to the sugar syrup in 3–4 steps, stirring until a white suspension is obtained, in which thickeners are added in the form of white clay and arabinogalactan, thoroughly mixing to obtain a composition in the form of a stable paste, of a light beige color, in following ratio of components, wt. %: ivermectin – 2.0 %, polyethylene glycol-400 – 7.0 %, polyvinylpyrrolidone-17 – 3.0 %, propylene glycol – 33.0 %, sucrose – 8.0 %, purified water – 12.0 %, sodium alginate – 0.3 %, edible white clay – 14.7 %, arabinogalactan – 20.0 %. Agent for the treatment of solid-hoofed animals with parasitosis is a thick sticky paste of light beige color, containing 2.0 % of ivermectin and 98.0 % of forming components. For ease of use, the paste is packaged in disposable syringes-dispensers with a capacity of 10–20 ml, designed for treatment of animal weighing 500–800 kg. Treatment and prophylaxis of solid-hoofed animals (horses, donkeys, mules) with nematodosis and botts of the digestive tract is carried out by agent obtained by the method described above, where the agent is administered once orally by a syringe dispenser along the toothless edge to the root of the tongue in a therapeutic dose of 0.2 mg/kg based on active substance or 1 ml of paste per 100 kg of animal weight.EFFECT: group of inventions provides an increase in the antiparasitic efficacy of the agent due to the formation of a soluble intermolecular complex, which delivers the drug to the parasite's organism during feeding, as a result of which the nervous system and the carbohydrate metabolism of the parasite are blocked, which causes the parasite to die.3 cl, 6 ex, 6 tbl

Description

The invention relates to agriculture, in particular to veterinary parasitology for the treatment of one-hoofed animals (horses, mules, donkeys) with helminthiases and entomoses.

Up to 100 species of various helminths have adapted to parasitization in horses, which under certain conditions of localization and intensity of invasion cause pathological processes in the body. Parasitizing in various organs and tissues of animals use it as a habitat, nutrition and reproduction of the population. They cause great harm to the definitive host, as well as during the migration of larvae in animals, can serve as a gateway to infectious diseases. The most widespread helminths of the Nematoda class are parascarides (Parascaris equorum) of the Ascaridae family, Strongylata of the order of 50 species, (Alfortia edentatus, Delafondia vulgaris, Srongylus equinus) cause severe diseases, under certain conditions with a fatal outcome, trichonematids - 37 species, oxiuris.

The ubiquitous chronic diseases of single-handed gastrophiliasis caused by the larvae of various types of gastric gadfly belonging to the genus Gastrophilus and the family Gastrophilidae also have pathogenic effects.

According to V.M. Ivashkina, G.M. Dvoinos [5, 6] V.A. Bolshakova [3, 4], L.A. Bundina [2], N.M. Ponamareva [8], D.A. Smirnova [9], B.E. Aituganova [1] and other researchers in certain regions, up to 90.0% of horses of all ages are infested with parasites, but young growth is more susceptible to infection, is more severely sick, and mortality is noted. Usually, mixed invasion occurs, which exacerbates the pathogenic effect on the animal organism, and mortality is noted. As a result, horse breeding causes great economic damage.

Currently, to combat parasitic diseases, drugs from the group of avermectins have become widespread. The fermentation product of the fungus Streptomyces avermitillis is a white crystalline substance, readily soluble in organic solvents and practically insoluble in water. In practice, drugs are used containing 1.0% active ingredient (DW) in a mixture of glycerol, polypropylene glycol, polyvinylpyrrolidone, polyethylene glycol or ethanol. The mechanism of action is that they stimulate the presynaptic secretion of gamma-aminobutyric acid (GABA), a neurotransmitter for inhibition. Exposed to the effects of ivermectin, nematodes and ectoparasites lose their ability to centrally control muscle activity - they paralyze and die. Compounds from the group of avermectins (ivermectin, aversectin, moxidectin, etc.) have high biological activity against nematodes and are 10 times higher than the known anthelmintics in activity. The dose of avermectins for animals is 0.2 mg / kg body weight, the drugs have nematodicidal, insecticidal and acaricidal effects.

When testing injectable forms of ivermectin P.A. Karns, D.G. Luther (1984) reported side effects in 366 of 3,316 horses, which was manifested by edema of the ventral abdominal wall in 10% of treated horses. 15 horses had edema at the injection site, 11 had swelling of the limbs, 4 had swelling of the eyes, 3 had colic, 2 had fever and one horse fell [11].

In connection with the manifestation of side effects in horses due to individual sensitivity and a special type of nervous system, injection forms of avermectins are almost not used. Forcible delivery of dosage forms in the form of suspensions or solutions is unacceptable, since they can enter the lungs due to the peculiarities of the horse's nasopharynx. Therefore, for oral administration to horses, ivermectins began to be used in the form of a paste. However, their use for mass deworming is constrained by the high cost.

To treat parasitosis of horses, domestic and foreign antiparasitic drugs based on ivermectin are used, these are injection solutions and pastes for horses [7, 8, 9, 10, 11, 12].

Ivermek - CJSC Nita-Farm, (Saratov, Russia) 1% injection solution with the addition of 4% vitamin E.

Side effects. With increased individual sensitivity to ivermek, in some animals, excitation, increased salivation (in dogs vomiting), increased bowel movements and urination, ataxia are observed. Weakened, depleted and sick with infectious diseases animals, as well as females in the last third of pregnancy, should not be treated. Collie and sheltie dogs are especially sensitive to the drug, a lethal outcome is possible.

Pasta Alezan - production of NEC "Agrovetzashchita (AVZ), Russia". The preparation contains 10% praziquantel and 2.0% ivermectin, as well as auxiliary components (sucrose, Santoquin, sorbitol, polyvinylpyrrolidone, gelatin and distilled water). A syringe dispenser in the amount of 6 g, at a price of 520 rubles. The disadvantage of the drug is the high price for mass deworming (100-500) horses. Substances from the group of avermectins and praziquantel in the composition of the paste are not dissolved, which up to 50.0% are excreted into the environment with feces, since avermectins adversely affect the bugs of coprophages and microorganisms involved in the decomposition of biological mass. Pastures are contaminated with undecomposed fecal matter during the grazing season.

Equisect paste - manufactured by Farmbiomed, Russia, active substance aversectin C. The drug contains 1% of the active substance aversectin C. The drug is given orally once at a rate of 0.23 mg / kg animal weight (2 g of paste per 10 kg of animal weight), syringe 14 g at a price of 300 rubles.

The disadvantage of the drug is that for the treatment of one horse weighing up to 600 kg, it is necessary to use 3-4 syringes with the drug in the amount of 900-1200 rubles. It is not convenient to introduce horses up to 56 g of paste and is not economically beneficial for mass deworming.

Imported anthelmintic pastes for horses:

Equalan - paste (Equalan, France), manufactured by MERIAL. As the active substance contains 1.87% ivermectin. Syringe dispenser 6.42 g at the price of 994 rubles. The drug for mass deworming of horses is not economically viable.

Equalan Duo (Equalan Duo, France) - paste, as the active substance contains 15.5 mg / g ivermectin and 77.5 mg / g praziquantel.

Auxiliary components: titanium dioxide - 0.155 g, antioxidant (E 320) - 0.02 g, dye (E 110) - 0.03 g and glycerin - up to 7.74 g. Equalan is released in bulk, disposable polypropylene syringes 6.42 g , 1625 rub.

The disadvantage of the drug is its high cost, also the auxiliary components E320 and the dye E110 are inherently carcinogens, and the latter can cause allergic reactions, in addition, side effects are: urticaria, rhinitis, chromosomal damage, swollen kidneys. And also a dietary supplement can cause vaginal pain in the stomach.

Intermectin Duo Paste (Intermedin Duo Paste, Canada) Contains in 1 g: ivermectin 15.5 mg; praziquantel 77.5 mg; 1 g adsorbed carrier. It is used orally in a dose of 0.2 mg invermectin and praziquantel 1 mg per 1 kg of body weight, respectively 1.29 g of paste per 100 kg of body weight with a single use. Each applicator containing 7.74 g of paste to process 600 kg of body weight. Price 1150 rub.

Side effects. Due to the use of Intermedin Duo Paste in some horses, swelling and irritation of the mouth, lips and tongue and salivation are noted.

Bimectin Paste (Bimectin Paste, Cambridge, Ontario, Canada.). A pale yellow paste containing 1.87% of ivermectin for oral use, 6.08 g in each syringe, for deworming of one horse weighing 570 kg, which corresponds to 200 μg of ivermectin per kg of animal weight. Price: 735 rub.

After using Bimectin (ivermectin) in horses, edema and itching are possible requiring symptomatic treatment.

Invest gel 2% (FORT DODGE Animal Health - USA), 1 ml of the drug contains the active substance moxidectin - 19 mg, and in 1 syringe 241.5 mg of moxidectin, a drug of prolonged action, protects horses from re-invasion by ecto- and endoparasites for 3 months with stall and grazing. Dosage, 0.4 mg / kg moxidectin per kg body weight, 1 syringe worth 1,450 rubles. designed for a horse weighing 525 kg. slaughter for food purposes is allowed 3 months after the last injection.

Disadvantages of macrocyclic lactones (avermectins):

1. individual sensitivity, in some animals (Collie shepherd dogs, thoroughbred and sport horses) after deworming side effects and even death are noted.

2. Preparations from the group of avermectins in the digestive tract of horses (cecum) reduce the number of beneficial intestinal microflora and simple single-celled organisms, which play a large role in completing the digestion process. Animals receive more than 65% of a complete protein of microbial origin and 70.0% of protozoa protein. In addition, microorganisms synthesize B vitamins (thiamine, riboflavin, nicotinic acid, folic acid, biotin, cyancobalamin, etc.).

3. Drugs are not removed from the body for a long time with a prolonged action of 14 to 32 days, they are well prescribed for young unproductive animals in the summer period of intensive parasite infestation, they are not given to productive animals, since milk and meat products cannot be used for food during the indicated period.

4. Feces from deworming animals in the pasture during the grazing season do not decompose, since part of the preparation with feces entering the external environment has a detrimental effect on beetles of coprophages and microorganisms. Pastures are littered with undeveloped feces with grass-free bald spots.

5. Injection preparations based on ivermectin cause severe pain and tissue necrosis at the injection site for a long time, non-absorbable seals appear, sometimes goats die from pain shock.

However, preparations based on macrocyclic lactones are very popular in veterinary medicine, as active antiparasitic drugs with a wide spectrum of action on endo- and ectoparasites and researchers are working to eliminate the noted deficiencies.

The aim of the present invention is to develop a highly effective, safe for use antiparasitic agent economically acceptable for mass deworming of domesticated, thoroughbred and sport horses.

The essence of the invention lies in the development of a highly effective ivermectin-based agent for veterinary practice with targeted additives that can have a positive effect on the removal of side effects, since this is important for veterinarians who are cautioned to deworm the thoroughbred and sports horses due to the possible manifestation of a side effect. Of no small importance is the cost of the drug, when processing a large number of horses, as well as the prevention of environmental pollution by chemical preparations excreted in feces.

The proposed tool consists of the active substance ivermectin and target additives: polyethylene glycol-400 (PEG-400); low molecular weight polyvinylpyrrolidone-17 (PVP); propylene glycol; sucrose chda; purified water; sodium alginate; arabinogalactan (AG).

The technical result of the invention is to increase the antiparasitic effectiveness of the drug due to the formation of a soluble intermolecular complex, which ensures the delivery of the drug to the parasite when it is fed, as a result of which the nervous system and the carbohydrate metabolism of the parasite are blocked, causing its death. Economic efficiency is ensured by the use of domestic inexpensive fillers and for the convenience of introducing the paste to animals in use as a dispenser of polyethylene disposable syringes with a capacity of 10-20 ml.

A distinctive characteristic of the paste developed by us is that:

- revealed the solubility of ivermectin in polyethylene glycol 400 when heated to 40-50 ° C.

- the nutrition of helminths was taken into account when parasitizing in the digestive tract, they needed carbohydrates, and for maximum delivery of the drug to the parasite's body, a soluble intermolecular complex of ivermectin (DV) substance with polymer excipients (PVP and AG) was added to the syrup from the sucrose and thoroughly mixed. In this case, targeted delivery of drug molecules to parasites of Drug Delivery Systems (DDS) occurs. As a result, carbohydrates with an integrated complex entering the body of the parasite during nutrition block the nervous system and carbohydrate metabolism, causing parasitic paralysis. This is an effective method for improving drugs using intermolecular / supramolecular complexes of drugs with oligosaccharides - cyclodextrins (CD) of the "guest-host" type.

- in the antiparasitic paste, the active substance ivermectin is in a soluble form compared to prototypes, providing better bioavailability to tissue parasites and larvae.

The listed positive features of the means, in accordance with the technical solution to the criterion of "novelty" allows us to conclude that the technical solution meets the criterion of "significant differences".

Choosing the components and mixing them in different ratios, we found the optimal pharmacopoeial composition required for the preparation of the dosage form in the form of a paste, which took into account: consistency; smell; taste; pH stability and safety.

The preparation according to the invention is illustrated by the following examples.

1. Polyethylene glycol (PEG) is an additive in food E-1521, which is used as a moisture-retaining substance, antifoam. The main properties of polyethylene glycol are an increase in the solubility of substances and their concentration [12].

2. Polyvinylpyrrolidone (PVP) is used as a binder in many pharmaceutical tablets, it is easily soluble in water and alcohol, and also due to the formation of water-soluble complexes it improves the solubility and bioavailability of chemotherapeutic agents [13].

3. Sodium alginate - a polysaccharide of natural origin, the chemical structure is a salt of alginic acid. It has valuable quality to impart viscosity and structure to liquid media. As a sodium thickener, alginate is also used in the production of processed cheeses, ketchup, ice cream, cottage cheese, canned meat, food additive E-401 [14].

4. Kaolin or white clay. Due to the content of calcium, magnesium and aluminum, it has a beneficial effect on the whole body. It is used not only for the treatment of a wide range of diseases, but also for cosmetic purposes.

5. Arabinogalactan “Lavitol (dihydroquercetin)” - the galactan-containing polysaccharide of higher plants is an immunomodulator. Extends shelf life. An excellent stabilizer and thickener, used to increase durability. Significantly reduces toxicity and increases the effectiveness of drugs. It has an overall high biological activity: hepatoprotective, antimutagenic, mitogenic - it stimulates the reproduction of spleen and bone marrow cells, has a noticeable gastroprotective effect, is an immunomodulator. Arabinogalactan has the properties of a probiotic, selectively stimulating the growth and activity of normal intestinal microflora, being food for bifidobacteria and lactobacilli (probiotics). It has high membranotropic activity, which provides targeted drug delivery, so it can be used to increase the absorption of drugs with low bioavailability [10, 15].

6. Propylene glycol is a colorless organic compound, almost odorless, transparent, viscous with a mild sweet taste. Food propylene glycol is used as an additive E-1520, which is a solvent and preservative of food products, and is also used in animal feed [16].

7. Sodium salt of carboxymethyl cellulose (Na CMC) - is used in the food and pharmaceutical industries as an emulsifier, stabilizer and thickener. Being inherently an insoluble polysaccharide, that is, dietary fiber, it is able to stimulate the gastrointestinal tract. In the manufacture of medicines, CMC is especially important when creating a variety of ointments, because it gives them the desired consistency and allows you to "keep in shape" for a long time. From the point of view of health, the main advantage of CMC is its neutrality and harmlessness [17].

Examples of specific performance:

Example 1. Preparation of the drug at the rate of 100 g of the finished product. The components of the drug are placed in a water bath in a certain sequence in compliance with these conditions.

Recipe 1. A mixture of polyethylene glycol-400 - 7 g and propylene glycol - 20 g was heated in a water bath to 50 ° C, 1 g of PVP-17 powder was added, thoroughly mixing with a mixer until complete dissolution (5 min.), Then 2 g of ivermectin was added with constant stirring for 5 min until the latter is dissolved, which is noted for the first time. The result is an intermolecular complex of ivermectin based on soluble polymers. An additional 40 g of propylene glycol and 3 g of sodium alginate were added to this solution to increase the weight of the components. To the resulting suspension, 27 g of white edible clay was added as a thickener while stirring until a paste of acceptable composition was obtained.

The ratio of the components of PEG-400; propylene glycol; ivermectin; Na alginate; white clay, respectively: 7: 60: 1: 2: 3: 27. The resulting product contains a large amount of propylene glycol 60 g per 100 g of product mass, this can have a harmful effect on animals, and may serve as a remark when registering the drug, so it was replaced by NaKMTS.

Recipe number 2.

The 1st stage 7 g of PEG-400 and 30 g of propylene glycol are heated to 50 ° C in a water bath or on an electric stove in an enamel pan, 3 g of PVP-17 powder is added to this solution, stirring with a mixer for 5 minutes until completely dissolved, in this mixture is added 2 g of the powder of the substance ivermectin stirring until it dissolves (5 min.). We obtain a soluble complex of ivermectin based on water-soluble polymers. (PEG-400, propylene glycol, PVP-17, ivermectin - 7.0: 30.0: 3.0: 2.0 - 42%).

2nd stage Pour 18 g of water into a separate dish, heat to 50 ° C and add 2 g of NaKMTs, mix until a stable suspension is obtained (water, NaKMTs, white clay - 18.0: 2.0 - 20%).

Stage 3 The resulting suspension of NaKMTs is added to the solution of the polymer-based ivermectin complex with a mixer. The result is a white precipitate of ivermectin partially precipitated, which is not desirable. When a thickener in the form of white clay 38 g is added, gas bubbles form in the paste, which must be removed in industrial production using vacuum (white clay - 38.0%). Therefore, NaKMTS, as a thickener and emulsifier, was replaced with sodium alginate, and water with sugar syrup, upon addition of which a stable suspension is formed.

The ratio of the product according to the recipe No. 2 PEG-400; propylene glycol; PVP-17; ivermectin; water; Na CMC; white clay is: 7.0: 30.0: 3.0: 2.0: 2.0: 38.0.

Recipe 3.

1st stage. Separately, we prepare sugar syrup in an enameled bowl, in which we fill 12 g of water, heat it in a water bath to 50 ° C and pour 8 g of sucrose and 0.3 g of sodium alginate into it, mix for 10 minutes to obtain a syrup. If water has evaporated, add water to 20 g (water, sucrose, sodium alginate: - 12.0: 8.0: 0.3).

2nd stage. Separately, we prepare a solution of the ivermectin complex. Pour 7 g of PEG-400, 33 g of propylene glycol into enameled dishes and add 3 g of PVP-17 powder at a temperature of + 50 ° C, mix with a mixer for 5 minutes until complete dissolution, then add 2 g of ivermectin, and mix until a homogeneous solution is obtained. The result is a solution of the polymer-based ivermectin intermolecular complex (PEG-400, propylene glycol, PVP-17, ivermectin: - 7.0: 33.0: 3.0: 2.0).

3rd stage. In the resulting solution of the molecular complex of ivermectin, we add the product of the 1st stage (sugar syrup with sodium alginate) in 3-4 doses. As a result of mixing, a stable suspension of white color is formed. Thickeners in the form of white clay and arabinogalactan (14.7: 20.0) - 34.7% are added to the resulting suspension of the product of the third stage, with stirring, to obtain a composition in the form of a persistent paste, light beige.

The ratio of the product according to the recipe No. 3 sugar syrup - (sucrose, water and sodium alginate - 8: 12: 0.3) - 20.3%; polymer-based ivermectin complex intermolecular solution - (PEG-400; propylene glycol; PVP-17; ivermectin - 7.0: 33.0: 3.0: 2.0) - 45.0%; thickeners - (white clay, arabinogalactan - 14.7: 20.0) - 34.7%, respectively, in%: 20.3: 45: 34.7%.

When developing paste No. 2 recipe, we noted that when replenishing the evaporated water, preparing a solution from the complex (PEG-400, propylene glycol, PVP-17, ivermectin), ivermectin partially precipitated. To test the paste prepared according to the recipe No. 3 for the stable solubility of DV, 100 ml of water and artificial gastric juice were poured into measuring cups, 1 g of paste was added and mixed thoroughly. The result is a stable suspension without sediment.

Thus, the resulting paste according to prescription No. 3 corresponds to the dosage form that we aimed for, where there is a stable consistency, when administered orally, unlike other pastes, it sticks slightly and spreads over the oral cavity and the animal has to eat it, has a light coniferous smell, sweet taste, stable preservation, the substance of ivermectin does not precipitate and does not delaminate during storage. In g of paste contains 20 mg of ivermectin (DW).

Antiparasitic paste No. 3 is a thick sticky texture of light beige color, contains 2.0% ivermectin and 98.0% of the forming components. In laboratory conditions, you can cook up to 5 kg of pasta. For ease of use, the paste is packaged in adapted disposable dosing syringes with a capacity of 10-20 ml.

In order to freely fill the syringe with paste, the cannula at its end is expanded with a drill, and the needle attached to the syringe is broken off at the base and ground. After filling the syringe with the paste to the desired volume, we use the polished base of the former needle as a sealing cap.

Industrial applicability

The technological process for the production of antiparasitic paste consists of two stages, based on the preparation of a 40% solution of sucrose in purified water. Further, in one mixer, by preparing a solution of ivermectin and sequentially adding other components with gradual stirring until a homogeneous paste is formed.

Based on the preparation of 100 kg of the finished product, the components of the drug are laid in the reactor in a strictly defined sequence in compliance with these conditions.

1. In reactor No. 1, sucrose, purified water for the preparation of 40% sugar syrup and sodium alginate are placed. Then stirred at a temperature of about 50 ° C until the sugar is completely dissolved. Download is carried out in accordance with the issued number of components and taking into account Krach + 1.004 (coefficient for drug losses).

Scheme for the preparation of sugar syrup (stage 1)

2. Polyethylene glycol 400, propylene glycol, polyvinylpyrrolidone are placed in reactor No. 2 and stirred at a temperature of not more than 50 ° C until complete dissolution. Then add the substance of ivermectin and continue to mix until the latter is dissolved. The heating of the reactor is stopped and gradually, in 4 doses, the sugar syrup obtained in the first reactor is added with constant stirring. A stable white suspension forms. Lastly, with stirring, white clay and arabinogalactan are added in 3-4 doses. Dissolution of the components occurs with stirring and a temperature of 40-50 ° C. Further mixing takes place at a temperature of 20-40 ° C for 20-30 minutes until a homogeneous plastic paste of light beige color is obtained.

Obtaining a solution of ivermectin (stage 2)

Further testing work was carried out with antiparasitic paste obtained according to the recipe No. 3 under the code name Equiverm.

Preclinical and production testing of ivermectin-based antiparasitic agents (Equiverm).

Pharmacotoxicological studies of equiver paste were carried out according to the “Guidelines for the preclinical study of the reproductive toxicity of pharmacological agents”, approved by the pharmacological committee of the Ministry of Health of Russia (protocol No. 8 of July 3, 1997) and approved by the Ministry of Health of Russia on December 18, 1997 [7].

Example 2. Acute oral toxicity was studied in rats and mice of both sexes weighing 160-180 g and 18-20 g, respectively. For each dose, 10 mice and 6 rats were used.

The paste was administered once with a gastric tube in doses of 5000; 10000 and 15000 mg / kg.

As a result, no deaths were recorded in any experimental group; moreover, no symptoms of intoxication were observed over the entire observation period (14 days).

It should be noted that a dose of 15000 mg / kg was administered to animals in two stages in equal portions with an interval of not more than 10 minutes. Immediately after administration, temporary inhibition was observed in animals, associated with a large volume of the substance administered. A dose of 15,000 mg / kg is the maximum possible for oral administration to rats and mice of both sexes. In this regard, the LD _{50 of the} tested paste will exceed this dose.

According to acute oral toxicity, the paste belongs to hazard class 4, according to GOST 12.1.007-76, and its LD ₅₀ will be more than 15000 mg / kg. The drug does not have species and sexual sensitivity.

Despite the fact that the equiverm is intended for oral use, for the safety of personnel working with it, it is necessary to evaluate its effect on the skin.

Example 3. The parameters of acute cutaneous toxicity were performed on 24 male rats weighing 200-220 g. For each dose, 6 rats were used, guided by the "Methodological guidelines for the formulation of studies on the study of irritating properties and substantiation of the maximum permissible concentrations of selectively acting irritating substances in air and working zone ", 1980. Equiverm paste was applied to the clipped part of the skin in doses of 2000; 5000 and 12500 mg / kg. Symptoms of intoxication and death of animals were not recorded. A dose of 12500 mg / kg was applied in equal portions in two stages, since it is the maximum possible for application to the skin.

In this regard, the LD _{50 of the} paste when applied to the skin is more than 12500 mg / kg. With a single application, the paste does not cause irritation. LD ₅₀ when applied to the skin is more than 12500 mg / kg, which corresponds to hazard class 4 according to GOST 12.1.007-76.

The paste does not show irritation when applied once to rat skin (0 points).

Example 4. The study of the pharmacokinetics of antiparasitic paste.

In the Moscow region, 5 horses were selected to study the pharmacokinetics of antiparasitic paste in blood plasma, which were administered oral antiparasitic paste from a dosage of 0.2 mg / kg of animal weight and then blood was taken from the jugular vein after 1; four; 8; 12; eighteen; 24; 36; 48; 60; 72; 84; 96; 108; 120 hours with a volume of 20 ml for the subsequent determination of ivermectin in the blood plasma of horses, by high pressure liquid chromatography, with ultraviolet detection using the method of R.K. Sanyal (1994).

A validated method for the determination of ivermectin in the blood serum of horses has been developed that allows this analyte to be determined in very low concentrations (LOD value is 0.03 ng / ml, LOQ is 0.09 ng / ml).

According to the results of the analysis of the samples, ivermectin is absorbed in the digestive tract of horses after oral administration and reaches determined concentrations in the blood serum within 1-2 hours after oral administration of antiparasitic paste - about 0.2 ng / ml. The maximum concentration of 0.84 ng / ml was recorded after 7 hours, and by 4 days the level of ivermectin gradually decreased to 0.2 ng / ml.

Thus, our results showed that after oral administration of ivermectin-based antiparasitic paste, it is absorbed quite well and enters the systemic circulation, where it circulates for 4 days. Two weeks after the use of the drug, only trace amounts of ivermectin are determined.

Example 5. The effect of antiparasitic paste on the body of horses

To determine the effect of antiparasitic paste on the horse's organism, 3 groups of five horses each were formed. The first group of horses was injected with a dose of 0.2 mg / kg; 2nd - 0.6 and 3rd - 1.0 mg / kg.

The clinical condition and blood sampling for hematological and biochemical studies were performed before and after 1; 3 and 5 days after giving the drug.

Thus, when studying the effect of antiparasitic paste on the body of horses, it was found that in a therapeutic, 3 and 5 times increased dose on the indicators of the clinical condition of horses there was no negative effect. Temperature, pulse and respiration were within the normal range and did not differ from the animals of the control group, and also did not adversely affect the hematological and biochemical parameters of the blood serum of horses.

Clinical Trials of Equiverm Antiparasitic Paste

For clinical trials, disposable dosing syringes with antiparasitic paste, 10 ml in volume, were prepared for the treatment of 300 horses with a live weight of 500 kg.

Example 6. Experience 1. Testing the effectiveness of a prototype antiparasitic paste was carried out in the Moscow region on 20 trotters aged 4-5 years.

To determine the degree of invasion of horses by nematodes, fecal samples were examined using the Fulleborn method using an ammonium nitrate solution to detect helminth eggs, while the eggs were diagnosed with a strong diagnosis of fever and feces were examined no later than one day. The eggs in g / feces were counted in a VIGIS counting chamber. For the diagnosis of oxyurosis, pieces of transparent adhesive tape (Scotch tape) were taken with imprints of perianal folds transferred to a glass slide and examined for the presence of oxyuride eggs.

For horses of experimental groups spontaneously infested with sorghilates, parascarides and oxyurides, the antiparasitic paste was administered orally along the toothless edge from a dosing syringe at a dose of 0.2 mg / kg in a single dose per 1 ml per 100 kg of animal weight (1 g of paste corresponds to 1 ml) .

In a preliminary study of 20 samples of feces of horses, eggs of strongilates were found in all samples on average 174.0 ± 8.7 ind. eggs / goal., in 8 young animals, parascaride eggs were found on average 347.1 ± 43.4 ind./ goal., eggs oxyurid was not found.

As a result, we obtained 100% effectiveness of ivermectin-based antiparasitic paste at a dose of 0.2 mg / kg against strongilates and parascarides.

Experience 2. In the second experiment, 54 outbred horses of different ages and sex in the Southern Federal District owned by a private entrepreneur were studied to determine the therapeutic dose of the drug.

The selection of animals in the experiment and determining the effectiveness of the drug was carried out according to the example of experiment 1.

According to the data of coproovoscopy of 54 studied samples of feces of horses, all eggs of strongilates were found on average 274.0 ± 54.88 ind. eggs / goal., in 20 young animals, parascaride eggs were found on average 447.1 ± 37.2 specimens / goal., in 21 animals, eggs oxyuride from 7 to 16 specimens. Of the horses invaded by Strongilyats, paraskarids and oxiurs at the same time, 4 experimental groups of 5 animals each were formed according to the principle of analogues. In the first group, equiverm paste was administered at a dose of 0.1 mg / kg; the second is 0.15; the third - 0.2 mg / kg; The 4th group for comparison (control) was injected with Equisect paste manufactured by Farmbiomed, Russia at the rate of 0.23 mg / kg animal weight (2 g of paste per 100 kg of animal weight) according to the instructions for its use.

As a result, Equiverm antiparasitic paste in doses of 0.1-0.2 mg / kg had 100% effectiveness against strongilates and hydroxyuris. Against parascarids at a dose of 0.1 mg / kg out of 5 animals, 3 were egg-free, extensivity was 60.0%, and the percentage of egg reduction was on average g / fec. amounted to 58.6%. At a dose of 0.15 mg / kg, out of 5 animals, 4 parascarids were completely freed from eggs (EE = 80.0%), and the number of eggs in g / fec. decreased to 99.7%. At a dose of 0.2 mg / kg, the drug was 100% effective against strongilates, parascarides and oxyuris.

Equisect at a dose of 0.23 mg / kg showed 100% effectiveness against oxyuris, against parascarides (out of 5 animals, 3 were completely released from eggs, EE = 60%), and the average number of eggs decreased from 460.3 to 22, 4 in g / fek. those. by 95.1%. Against strongilates (out of 5 animals, 4 strongilates were released, EE = 99.7), and the average number of eggs decreased from 280.5 to 0.8 in g / fec., By 99.8%

The main nematodes parasitizing in the digestive tract are almost always found in animals as a mixed invasion. Some of them show higher resistance to antiparasitic drugs. Since animals are infected in the form of a mixed invasion, and the equiver at a dose of 0.2 mg / kg showed 100% effectiveness against the main parasites of the digestive tract, we accepted it as a therapeutic one (the results are shown in Table 2, appendices) .

Experience 3. To confirm our data, a commission of leading scientists and veterinary specialists was created. The effectiveness of antiparasitic paste based on ivermectin in the commission experience is shown in table. 3, from which it can be seen that, according to the data of coproovoscopic studies of fecal samples, all 14 horses were invaded by strongylates with an average number of eggs per g / fec. ind., 652.4 ± 46.6 ind., of which 5 - parascarides - 315.2 ± 22.5 ind. and according to imprints from perianal folds in 7 samples, oxyur eggs were found on average 8.4 ± 0.6 ind.

After deworming with ivermectin-based antiparasitic paste at a therapeutic dose of 0.2 mg / kg recommended by us, calculated as 1 ml of paste per 100 kg of animal weight, 100% efficacy against strongilates, oxyuris and parascarides was obtained. Side effects after giving the drug are not marked (the results are shown in table. 3, applications).

Experience 4. To determine the effectiveness of the equiverm against the larvae of gadflies localizing in the digestive tract of horses, 20 horses were selected, which were divided into 2 equivalent groups.

The first group of animals was dewormed with equiverm paste at a therapeutic dose of 0.2 mg / kg, the second group served as a control and the drug was not received. A day after deworming, stools were collected from each horse of the first group and washed thoroughly in a bucket of water. As a result, from 7 to 10 samples, from 33 to 120 copies were detected. pinkish-red larvae of Gasterophilus spp. In washed feces from control animals, gastrophilus larvae were not found. Then the horses of the control group were dewormed with equiverm paste and from 8 feces 935 gadfly larvae were found per animal on average 116.9 specimens, which confirms the high extent-invasion (EI = 75.0%) of horses by gadfly larvae in the Southern Federal County and paste effectiveness equiverm against them. Since larvae of intestinal gadfly are not diagnosed according to coprooscopy, it is necessary to carry out preventive deworming in the summer during the invasion of horses by larvae of the gadfly (Gasterophilus spp) of the digestive tract.

The estimated cost of the components of the drug per 100 g are shown in table No. 4, from which it follows that the cost of the components for preparing 100 g of the drug is 94.5 rubles, 100 g of the drug is enough for 20 doses (100 ÷ 5 = 20 g) per 5 g (5 ml) of the drug per animal weighing 500 kg and it costs (94.5 rubles. ÷ 20 g) 4.7 rubles., plus a disposable syringe 5 rubles. As a result, the cost of one syringe dispenser designed to handle a horse weighing 500 kg is 9.7 rubles without commercial allowances, which is much lower than the cost of its prototypes.

Conclusion The antiparasitic paste developed on the basis of the substance ivermectin under the conditional name “Equiverm” for treating one-hoofed animals (horses, donkeys, mules) from larval and adult stages of the digestive tract parasites, is a thick sticky paste of light brown color, contains 2% ivermectin and 98, 0% forming components. The paste has a stable consistency, when administered orally, unlike other pastes, it sticks slightly and spreads over the oral cavity and the animal is forced to eat it, has a light coniferous smell, sweet taste, has good preservation, the substance of ivermectin does not precipitate and does not separate during storage . G of paste contains 20 mg of ivermectin (DV), which corresponds to 1 ml. For ease of use, the paste is packaged in disposable dosing syringes with a capacity of 10-20 ml.

Preclinical trials have shown that the drug belongs to hazard class 4, according to GOST 12.1.007-76, does not show irritating effects on the undamaged skin of rats, does not have species and sexual sensitivity, and embryotropic activity.

A study of pharmacokinetics showed that after oral administration of ivermectin-based antiparasitic paste, it is absorbed quite well and enters the systemic circulation, where it circulates for 4 days. Two weeks after the use of the drug, only trace amounts of ivermectin are determined. This allows 2 times to reduce the term of slaughter of animals for meat, in contrast to the equiverm prototypes, the metabolites of which ivermectin are excreted from the body of animals within 21-30 days.

When studying the effect of antiparasitic paste on the body of horses, it was found that in a therapeutic, 3 and 5 times increased dose on the indicators of the clinical condition of horses, there was no negative effect, as well as the effect on the hematological and biochemical parameters of the blood of horses.

Clinical trials have shown that the drug at a dose of 0.1 mg / kg, in contrast to prototypes, showed 100% efficacy against paraskarids and gastrophylus larvae and oxiuris and 0.2 mg / kg. The commission test of the equiverm in the therapeutic dose of 0.2 mg / kg we have established has confirmed its high efficiency and safety of use against the main parasites of horses.

The cost of a dosing syringe with Equiverm antiparasitic paste, calculated on the processing of an animal weighing 500 kg, is 9.7 rubles without surcharges, which is much cheaper than prototypes.

Literature.

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application

Recipe 2.

Recipe number 3

Claims (4)

1. A method of obtaining funds for the treatment of one-animal animals with parasitosis, comprising three stages, in the first stage, sugar syrup is prepared in a separate bowl by heating water to + 50 ° C, with stirring with the addition of sucrose and sodium alginate, in the second stage in another bowl with at a temperature of + 50 ° C, a solution of the ivermectin intermolecular complex based on polymers, PEG-400, propylene glycol, PVP-17 is prepared, in the third stage, the ivermectin solution is added to sugar syrup in 3-4 doses, stirring until a stable white suspension is obtained in which thickeners are added in the form of white clay and arabinogalactan, mixing thoroughly to obtain a composition in the form of a persistent paste, light beige in the following ratio of components, mass. %: Ivermectin  2.0 PEG-400  7.0 PVP-17  3.0 Propylene glycol  33.0 Sucrose  8.0 Purified water 12.0 Sodium Alginate  0.3 White clay 14.7 Arabinogalactan  20,0 2. A tool for the treatment of one-animal animals with parasitosis, obtained by the method according to claim 1, where the paste for dosed administration to animals is packaged in adapted disposable syringes with a capacity of 10-20 ml, designed to handle an animal weighing 500-800 kg. 3. A method for the treatment of one-animal animals with nematodoses and larvae of the digestive tract gadgets by the agent obtained by the method according to claim 1, wherein the agent is administered orally once with a syringe-dispenser along the toothless edge to the root of the tongue at a therapeutic dose of 0.2 mg / kg according to the active substance or 1 ml of paste per 100 kg of animal weight.