RU2219913C2 - Anthelminthic preparation - Google Patents

Abstract

FIELD: agriculture, veterinary helminthology. SUBSTANCE: the suggested innovation deals with treating nematodoses in animals. Anthelminthic preparation includes active substances as levamisol and oxphendasol in decreased dosages and supplementary surface-active substance as low-molecular polyvinyl pyrrolidone (mol. weight 10-15 thousand) at the following ratio of components: 2.5 : 1.5 : 0.25. Preparation at therapeutical dosage of 4.5 mg/kg is of high (98.6-100%) anthelminthic activity at strongylatoses of gastrointestinal tract and dictyocaulosis of small cattle and canine toxocariasis. Anthelminthic preparation is of no embryotoxic and teratogenic properties. Its LD₅₀ for mongrel white mice at peroral intake corresponds to 450 mg/kg, for white rats - 700 mg/kg. EFFECT: higher efficiency. 5 tbl

Description

 The invention relates to the field of agriculture, namely to veterinary helminthology, and can be used to treat nematodoses of animals.

 Of the known agents for the treatment of animal nematodoses, piperazine, naphthamone, thiabendazole, tetramisole, fenbendazole, mebendazole, albendazole (I) are widely used.

 The disadvantages of these drugs are high therapeutic doses and a narrow spectrum of anthelmintic action, high toxicity, the presence of embryotropic properties, their deficiency.

 Closest to the claimed technical solution are levamisole and oxfendazole, which are used for strictilatosis of the gastrointestinal tract and dictation of lung lungs of sheep in doses of 7.5 and 4.5 mg / kg, respectively.

 The disadvantages of the drugs include high general toxicity in levamisole (2) and specific toxicity (presence of embryotropic properties) in oxfendazole (3). In addition, oxfendazole does not dissolve in water, is not wetted by water and does not form a suspension.

 The proposed anthelmintic agent includes active substances - levamisole and oxfendazole - in reduced doses and an auxiliary surfactant - polyvinylpyrrolidone (PVP) with a molecular weight of 10-15 thousand - with a ratio of components of 2.5: 1.5: 0.25 .

 According to the mechanism of anthelmintic action, levamisole and oxfendazole are synergists that affect the energy processes of helminths: levamisole inhibits glycolysis, and oxfendazole affects enzymatic processes. Ultimately, paralysis of the muscles of the helminths occurs and their death.

 Polyvinylpyrrolidone is a surfactant, detoxifier and has a prolonging effect (4).

 A comparative comparison of the proposed technical solution with the prototype shows that the claimed tool is characterized in that the active substances levamisole and oxfendazole are taken in lower doses, which make up one third of the therapeutic dose of the substances, and additionally contains an auxiliary surfactant, low molecular weight polyvinylpyrrolidone.

An anthelmintic agent in a therapeutic dose (4.25 mg / kg) and 3 times increased (12.75 mg / kg) does not have an embryotropic effect. LD ₅₀ for white mice 450 mg / kg. In sheep and dogs, the drug in 3- and 5-fold therapeutic doses (13.5-22.5 mg / kg) does not cause toxicosis.

 An anthelmintic agent is a white powder that is well wetted by water and forms a suspension solution with it.

 The drug is used for the treatment of strongilitis in ruminant and toxocariasis dogs at a dose of 4.25 mg / kg, i.e. on DV 4 mg / kg.

 In laboratory and industrial conditions, an anthelmintic agent is obtained by mechanical mixing of 2.5 parts of levamisole, 1.5 parts of oxfendazole and 0.25 parts of PVP.

 For these purposes, you can use mixers of various types, providing uniform mixing of the components.

 The drug is used in ruminants at a dose of 4.25 mg / kg (DV 4 mg / kg) in a mixture with concentrated feed or in the form of a 4% aqueous suspension (0.1 ml / kg), which is prepared on the eve of deworming.

 The drug can also be used in animals in the form of 3 and 2% suspensions (0.15 and 0.2 ml / kg, respectively).

Examples of specific performance
Example 1. The development of options for the ratios of active substances - levamisole and oxfendazole - and the determination of the synergism of their anthelmintic action in case of strongilatosis of the gastrointestinal tract of small cattle.

Several combinations of the ratios of active ingredients - levamisole and oxfendazole have been developed:
1) 4: 2.5 = 6.5,
2) 3: 1.875 = 4.875,
3) 2: 1.25 = 3.25,
4) 3: 2 = 5,
5) 2.5: 1.5 = 4,
which were tested for anthelmintic activity with strong digestive tract sheep and goats.

 Experiment 1. In the experiment, 25 lambs were used, which were naturally infested with strongilates of the gastrointestinal tract (nematodiruses and other strongilates).

 Each dose of the combined preparation (a total of three doses was tested: 6.5; 4.875 and 3.25 mg / kg in DV) was tested in 5 animals. In addition, for comparison, each drug was separately administered to 5 lambs in the recommended dose: levamisole at a dose of 7.5 mg / kg, oxfendazole 4.5 mg / kg. The effectiveness of anthelmintic agents in the tested doses was determined by helminthovoscopic examination of feces according to the Fulleborn method before treatment and 7-10 days after treatment. The number of eggs in 1 g of feces was taken into account.

 The results of the experiment are shown in table 1.

 As a result, it was found that the use of the first and second combinations of levamisole and oxfendazole (6.5 and 4.875 mg / kg) yielded a 100% therapeutic effect in case of strong gastrointestinal tract infections in sheep. When using the 3rd combination of drugs (3.25 mg / kg) of five lambs, 4 animals were completely cured of strongilates and one partially; IE = 99.9%.

 All animals were cured of strongilates from the use of oxfendazole at a therapeutic dose of 4.5 mg / kg (efficiency is 100%), however, when using levamisole at a therapeutic dose of 7.5 mg / kg (oral), four of five lambs were completely cured and one partially ; IE = 98.9%.

 Experiment 2. The experiment was conducted on 22 sheep and goats, naturally invaded by strongilates of the gastrointestinal tract (nematodirosis, etc.).

 The animals of the first experimental group (n = 5) were given the 4th combination of levamisole and oxfendazole (5 mg / kg), the second group (n = 5) - the fifth combination (2.5 + 1.5 = 4 mg / kg), the third group (n = 6) - levamisole at a dose of 2.5 mg / kg, the fourth (= 6) - oxfendazole at a dose of 1.5 mg / kg. The results of the experiment are shown in table 1.

 As a result of helminthovoscopic examinations of sheep and goats 10 days after treatment, it was found that 4 and 5 combinations in total doses of 5 and 4 mg / kg body weight, respectively, showed 100% efficiency. Levamisole and oxfendazole in reduced doses of 2.5 and 1.5 mg / kg, respectively, were ineffective.

 Thus, 4 mg / kg of the animal’s body weight, i.e. 2.5 parts of levamisole is taken 1.5 parts of oxfendazole.

 Example 2. The development of a prescription complex drug.

 In order to develop the optimal variant of the anthelmintic suspension, II suspensions were made, which included the established ratio of active substances - levamisole and oxfendazole - and various amounts of auxiliary surfactant PVP.

The best option is recognized as a prescription for a suspension of anthelmintic, including
levamisole - 2.5 g
oxfendazole - 1.5 mg
PVP - 0.25
water - up to 100 ml
those. 4.25 of the drug powder is suspended in water up to 100 ml.

 This suspension is characterized by the following indicators in a cylinder with a capacity of 50 ml with divisions: after 1 hour of sedimentation, the suspension is clarified by 1 division, i.e. per 1 ml; after 3 hours at 1.2 divisions; after 1 day into 3 divisions; after 3 days at 6 divisions. The sediment at the bottom of the cylinder after 3-day sedimentation of the suspension is completely resuspended after 10-12 unsharp shaking-transfusions of the suspension to the cork and back.

Thus, as a result of the development of the optimal variant of the suspension, it was found that the recipe of the dry powder of the drug includes
levamisole - 2.5 parts
oxfendazole - 1.5 parts
PVP - 0.25 parts
All components must be mixed in a mixer to obtain a uniform powder.

 Example 3. Tests of an anthelmintic agent in case of strongilatosis of the gastrointestinal tract of small cattle.

 Experience 1. In order to clarify the therapeutic dose for strictilatosis of the gastrointestinal tract of small cattle, the drug was tested at a dose of 4.25 mg / kg (DV 4 mg / kg). The experiment was conducted on 23 experimental animals with 7 control. The anthelmintic was given orally and to goats orally after overnight fasting in the form of a 4% suspension (0.1 ml / kg).

 The effectiveness of the drug was determined by the method of "critical test", taking into account the number of eggs in 1 g of feces before treatment and 7-14 days after treatment.

 The results of the experiment are presented in table 2. As can be seen from the table, the drug at a dose of 4.25 mg / kg with nematodirosis showed EE = 94.4%, IE = 98.6%, with other strictilitis (hematosis, ostertagia, bunostomosis) its effectiveness amounted to 100%.

 Experience 2. The production test of the anthelmintic agent was carried out in a therapeutic dose of 4.25 mg / kg (at DV 4 mg / kg) on 43 sheep and goats spontaneously infected with strongilates of the gastrointestinal tract (table 2). The drug was administered to animals in the form of a 2% suspension (0.2 ml / kg) orally on an empty stomach, after nightly fasting. After administration of the drug, no deviations in the physiological state of the animals were observed.

 An examination of animals for infection with strongilates after treatment was carried out after 7, 14 days.

 As a result, it was found that out of 43 treated animals, single eggs of nematodiruses and other strongilates were found only in 2 different samples of feces, all other animals were completely cured of strongilata of the digestive tract.

 Thus, an anthelmintic agent at a dose of 4.25 mg / kg (at DV 4 mg / kg) in the form of a 2% suspension solution (0.2 ml / kg) for individual oral administration has shown high efficacy in case of strong gastrointestinal tract infections tract of small cattle: with nematodirosis EE = 92.9%, IE = 99.5%; with other strongilitis (bunostomosis, hematosis, ostertagia) EE = 97.7%, IE = 99.93%.

 Example 4. Testing an anthelmintic agent for dictation of sheep.

 2 experiments were conducted to test the drug for dictation of sheep dictyocauliasis (table 3). Invasive animals intended for the experiment were detected from flocks using helmintholaroscopic examination of feces according to the Bormann-Orlov method in the modification of Shilnikov. The number of helminth larvae in 1 g of feces was taken into account.

 The first experiment was carried out on 9 sheep, divided into three equivalent groups of 3 animals each. The sheep of the two experimental groups were given individually an anthelmintic in the form of a suspension in the doses of 4.25 and 5.31, by mouth, i.e. for DV 4 and 5 mg / kg, and 3 sheep were left as a control (the drug was not administered).

 The effectiveness of the drug was determined by the method of "critical test", examining the animals for the presence of parasite larvae and their number before treatment and 10 days after application of the drug. As a result, it was found that all the sheep that were used in doses of DV 4 and 5 mg / kg were free from helminths (100% effective), while in three control sheep they found an average of 1 g of feces 19, 3 ± 3.5 dictiocaul larvae.

 The second experiment was carried out on 20 sheep spontaneously infected with diktiocaulus. In the animals of the first experimental group (n = 10), the drug was given in a similar way at a dose of 4.25 mg / kg (at DV 4 mg / kg), the second group (n = 5) at a dose of 5.3 mg / kg (at 5 mg / kg), the sheep of the third group (n = 5; control) the drug was not prescribed. 12 days after deworming, we examined all the sheep for dictiocauliasis.

 As a result, it was established: out of 10 animals of the first experimental group, only one sheep found single larvae of dictiocaulus in a feces sample (on average 0.2 specimens); in 5 animals of the second experimental group, no larvae were detected in samples of feces; in 5 sheep of the control group, the invasion intensity was 11.6 ± 2.3 ind. larvae on average per animal.

 Thus, the drug at a dose of 4.25 mg / kg (at DV 4 mg / kg) for dictation of sheep showed EE = 90%, IE = 97.88%; at a dose of 5.31 (for DV 5 mg / kg), its effectiveness was 100%.

 We took the dose of the anthelmintic agent 4.25 mg / kg (at 4 mg / kg DV) as the therapeutic dose for sheep dictation, which provides a high therapeutic effect (IE = 97.88-100%).

 Example 5. A commission test of the effectiveness of the drug in case of strictilatosis of the gastrointestinal tract and dictation of sheep.

 A commission test of anthelmintics was carried out in the Stavropol’s industrial holding, Stavropol Territory. Under the experiment, we took 20 ewes spontaneously invaded by nematodes of the gastrointestinal tract and lungs.

 Formed 4 groups, numbered and weighed the animals. Sheeps of groups 1-3 were individually individually orally administered, respectively, with an anthelmintic agent at a dose of 4.25 mg / kg (DV 4 mg / kg); levamisole in a dose of 7.5 mg / kg in terms of active ingredient (prototype) - and oxfendazole in a dose of 4.5 mg / kg in terms of active ingredient (prototype). The animals of the fourth group were not administered drugs and they served as a control.

 The actual results of the experiment are presented in table 4.

 As a result of the commission audit, the following was established. During helminth infections and larvoscopic examination of feces samples, 100% infection of animals with nematodes was established prior to drug administration.

 13 days after treatment, no dictiocaul larvae were detected in fecal samples of experimental sheep (EE = 100%). In case of strong gastrointestinal tract infections, the decrease in the rate of egg production was: in the group of animals treated with an anthelmintic, 99.12%, levamisole 100%, oxfendazole 94.75%. Moreover, in animals of the control group, an average of 282.6 ± 88.79% of eggs were registered.

 Thus, the results of the commission test showed a high anthelmintic activity of the anthelmintic agent with a single oral administration at a dose of 4.25 mg / kg (DV 4 mg / kg) of live weight in sheep spontaneously invaded by nematodes of the gastrointestinal tract and dictiocaulus.

 The effectiveness of the drug is not inferior to the prototypes - levamisole and oxfendazole.

 Example 6. Test anthelmintic in dogs with toxocariasis (table 5).

 The drug was tested at a dose of 4.25 mg / kg (DV 4 mg / kg) in 5 puppies spontaneously infected with toxocaras. In parallel, 5 animals were given the basic drug tetramisole at a dose of 10 mg / kg twice after 24 hours. Both drugs were given to puppies in a similar way by free-feeding in a mixture with minced meat.

 The effectiveness of autohelminthics was determined by taking into account departed toxocars within three days after treatment, as well as by examining the animals for the presence of nematode eggs in feces 7 days after treatment.

 As a result, it was found that the anthelmintic agent at a dose of 4.25 mg / kg (4 mg / kg in the DV) once showed 100% effectiveness in dogs with toxocariasis.

 Similar efficacy was obtained from the use of the prototype tetramisole at a dose of 10 mg / kg twice.

 Example 7. Determination of acute toxicity of an anthelmintic agent.

 The acute toxicity of the drug was studied in white mice and rats. The anthelmintic was administered in the form of a suspension orally once to mice in doses from 200 to 600 mg / kg in the DV, to rats from 450 to 1500 mg / kg of body weight. For each test dose, 6 animals were taken. After the introduction of the drug, the animals were monitored for 7 days, taking into account the time of the onset of poisoning symptoms and their nature. The timing of the death of animals was recorded.

The parameters of acute toxicity of the anthelmintic agent for outbred white mice were established: LD ₀ = 300.0; LD ₁₆ = 345.0; LD ₅₀ = 450.0; LD ₈₄ = 550.0 and LD ₁₀₀ = 600 mg / kg.

For white rats, LD ₀ = 450.0; LD ₅₀ = 700.0 and LD ₁₀₀ = 1500 mg / kg. The calculated data for LD ₅₀ were verified experimentally and were fully confirmed in both cases.

 The drug was more toxic to mice than to rats.

 According to the hazard classification of substances, an anthelmintic agent is classified as moderately hazardous.

 Example 8. Testing drug tolerance.

 The experiments were carried out on adult sheep. According to the principle of analogues, 3 groups were formed: 3 animals in each. The drug was administered in 3, 5 and 10-fold therapeutic doses, i.e. the anthelmintic was administered orally to sheep in the form of a suspension in doses of 12, 20 and 40 mg / kg body weight.

 It was established that 3 and 5-fold therapeutic doses of the drug do not cause visible signs of toxicosis, 10-fold doses cause slight short-term excitation in animals.

 Example 9. Determination of chronic toxicity of an anthelmintic agent.

The experiment was performed on 20 rats weighing 110-120 g. The drug was administered to animals orally at a dose of 1/10 of LD ₅₀ , i.e. 70 mg / kg over 2 months.

 Throughout the experiment, animals were monitored and weighed daily.

 As a result, the absence of a cumulative effect in the preparation was established, which is confirmed by the absence of rat death.

 Example 10. The study of the local action of anthelmintic agents.

 The experiment was conducted on 40 rats, divided into 2 equivalent groups. The first group received the drug in a therapeutic dose of 4 mg / kg in terms of DV, the second - in a three-fold therapeutic dose of 12 mg / kg in terms of DV. After 3, 6, 24 and 72 hours, 5 animals from each group were killed and the condition of the mucous membranes of the digestive tract was examined.

 Signs of inflammation were absent in all cases, which indicates the absence of an irritating effect on the drug.

 Example 11. The study of the embryotropic properties of anthelmintic drugs.

 The embryotoxic and teratogenic properties of the drug were studied in therapeutic (4 mg / kg DV) and triple therapeutic doses (12 mg / kg DV). For each dose, 36 female rats were taken, divided into 4 equivalent groups of 9 animals each.

 Group I received an anthelmintic agent during the period from 1 to 7 days of embryogenesis, II - from 8 to 15 days, III - from 16 to 19 days of embryogenesis.

 Group IV animals served as a control and did not receive the drug.

 On the 21st day of pregnancy, the rats were killed and the resulting embryonic material was examined. The number of corpus luteum pregnancy in the ovaries, implant sites, living, dead and ugly fruits, resorptions was calculated. The pre-, post-plantation and total death of the embryos, the size and weight of the fetus and placenta were determined.

 As a result of studies of embryotoxic and teratogenic properties in an anthelmintic agent in a therapeutic and triple increased therapeutic dose was not detected.

Sources of information taken into account when preparing the application
1. Instructions on the prevention and elimination of animal diseases with helminth infections. - M .: Informationgrotech, 1999 - 72 p.

 2. Fetisov V.I. Bull. All Institute of Helminthology. - M., 1974 - 13-14.

 3. Delatour P., Debroye I., Lorgue G., Courtot D. Embryotoxici.te experimentale de l'oxfendasole chez la rat et la mouton // Res. Med. Bet. 1977. - V.153, N 10. - P.639-645.

 4. Mashkovsky M. D. Drugs: In 2 volumes. T.2-11th ed. erased. - M .: Medicine, 1988. - P.103-104.

Claims (1)

Anthelmintic agent containing levamisole, characterized in that it additionally contains oxfendazole and a low molecular weight surfactant polyvinylpyrrolidone in the following ratio of components: levamisole: oxfendazole: low molecular weight polyvinylpyrrolidone - 2.5: 1.5: 0.25.

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