RU2699795C1 - Agent for treating eimeriosis in animals - Google Patents

Abstract

FIELD: veterinary science.SUBSTANCE: invention relates to veterinary science and represents an agent for treating eimeriosis in animals, containing N-(4-bromobenzylidene)octadecan-1-amine as an active substance and sunflower oil as a solvent in the following ratio: N-(4-bromobenzylidene)octadecan-1-amine : sunflower oil = 1:99.EFFECT: invention provides a wider range of products for treating eimeriosis in animals, high anti-parasitic effectiveness and low toxicity and low irritant action.1 cl, 3 tbl, 4 ex

Description

The invention relates to veterinary medicine, more specifically to parasitology, and in particular to agents for treating eimeriosis in animals containing (agent) N- (4-bromobenzelidene) octadecane-1-amine

as an active substance and sunflower oil, as a solvent at their weight ratio of 1:99, can be used as an anti-emeriosis (anticoccidiosis) drug in veterinary medicine for the treatment of farm and domestic animals. The claimed anti-eimeriotic agent is administered orally at a dose of 10 mg / kg of live weight of the animal twice with an interval of 6 days. The claimed tool has high antiparasitic efficacy and the absence of toxicity (LD50 2500 mg / kg), as a result of which it is practically safe when used as directed and provides an increase in the effectiveness of treatment.

It is well known that severe forms of parasitosis in animals are predominantly recorded against the background of immunodeficiency states. In the body of animals and birds with eimeriosis, intoxication occurs with the products of metabolism of parasites and putrefactive microflora with bloody secretions, which leads to impaired function of the nervous system, causing inhibition of animals and birds, up to a coma, muscle tremor and paralysis of limbs. Animals and birds lose weight, meat of ill animals has low nutritional value and quickly deteriorates during storage. In birds, egg production decreases.

Coccidiosis, namely eimeriosis, is an acute, subacute and chronically occurring disease of animals and birds caused by various species of protozoa belonging to the Protozoa kingdom and to the family Eimeriidae. Eimeriosis is caused by protozoa that are parasitic mainly in the intestinal epithelial cells. With a strong degree of invasion, eimeriosis can cause mass death of animals [Akbaev M.Sh., Vasilevich F.I., Akbaev R.M. et al. Parasitology and invasive animal diseases. - M .: Kolos, 2008, - 756 p.].

From the studied prior art, the applicant has identified a number of drugs with anti-emeriosis activity for the treatment of eimeriosis in animals and birds: furazolidone, furacryllin, furazolan (nitrofuran drugs), chemcocide, trichopol, salinomycin, baykoks, sulfanilazolezolinazolin, sulfadimethazole, sulfadimetolazin-azolethoxin ), as well as chloramphenicol, monomyecin and many others [Akbaev M.Sh., Vodyanov A.A., Kosminkov N.E. et al. Parasitology and invasive animal diseases. - M .: Kolos, 2000, - 559 p .; Pleshakov S.A., Larionov S.V., Rozovenko M.V. The effectiveness of furacryllin and furazonal in eimeriosis of rabbits. Veterinary medicine, Saratov, 1997. No. 8, p. 28-30].

However, currently available anti-eimeriosis drugs have one or more negative properties:

- weak anti-eimeriotic effect, high doses and repeated use, high price;

- high toxicity;

- cumulation in edible animal tissues;

- immunosuppressive effect;

- prolonged use in veterinary medicine and, as a consequence, the danger of the development of resistant strains of the pathogen.

Despite the widespread use of drugs as a treatment and prevention of eimeriosis, 100% indicators of the extent and interest effectiveness of the investigated prior art, the applicant has not been identified. Therefore, these drugs are used repeatedly. For example, to treat eimeriosis in rabbits, furazolidone is used at 250 mg / kg for 10 days, and after a 5 day break, sulfadimethoxine is used on the first day, 2 g, and in the next 9 days, 1 g per animal [Ponomarenko V.Ya. Materials of the scientific conference of parasitologists UAASSR. Kiev, 1975, - S.116.].

A disadvantage of the known technical solution is that the effective concentration of known drugs is much higher than the effective concentration (dose) used in the claimed technical solution, while the doses indicated in the known source are administered for several days - up to ten, and repeated treatment with other drugs is required, which, ultimately, leads to high costs of both material and labor resources.

From the studied prior art, the applicant revealed the invention according to the patent of the Russian Federation 2234932, the essence is a method of treating cows suffering from eimeriosis, including the administration of coccidin with food, characterized in that the animals are additionally injected subcutaneously once with a 10% solution of heraconda in a dose of 4.5-5, 5 mg / kg body weight.

The disadvantage of this method is the repeated use of the drug, namely, first, coccidine is administered orally, and then heraconda solution is injected, and the treatment is characterized by complexity and high cost.

From the studied prior art, the applicant revealed the invention according to the patent of the Russian Federation 2322235, the essence is a method of treating eimeriosis in animals and birds, including twice, with an interval of 5 days, feeding xymedon or xymedone hydrochloride in a dose of 30 mg / kg of body weight to small animals and additionally introducing furazolidone at a dose of 20 mg / kg body weight in a five-day course with an interval of 3 days.

The disadvantage of this method is the repeated use of two drugs in high doses.

Thus, the principle of using anti-eimeriotic drugs at the date of submission of application materials is based on their repeated use, which may have a negative side effect on the body (non-indifferent to the body).

Many of the known drugs have toxic, irritating and immunosuppressive effects. In addition, prolonged use of one drug leads to the appearance of dysbiosis, inhibits the enzymatic activity of the gastrointestinal tract, and can cause other functional changes in the body of birds and animals.

From the studied prior art, the invention was revealed according to the patent of the applicant of the Russian Federation 2473334 "Antiemeriose pharmaceutical composition based on the quaternary phosphonium salt and substituted dinitrobenzofuroxan", the essence is the antimeriotic pharmaceutical composition containing n-tetradecyltri-n-butylphosphonium chloride and 5,7-bis- (m- nitroanilino) -4,6-dinitrobenzofuroxan as active components with a weight ratio of 1:10 and glucose as an auxiliary substance. The composition is used by 2-fold administration with an interval of 24 hours at a dose of 4 mg / kg (in terms of the active substance - phosphonium salt) with LD ₅₀ 2500 mg / kg.

A disadvantage of the known technical solution is the need for double use of the drug. In addition, the well-known product is characterized by a rather complex and expensive synthesis, which does not contribute to its widespread use, while neither the drug itself nor its components are commercially available in the Russian Federation, because It is synthesized exclusively in the laboratory of Kazan Federal University, due to the fact that RF patent No. 2473334 belongs to KFU and has not been brought to the stage of industrial production due to lack of investment for preclinical and clinical trials.

The inventive patent of the Russian Federation patent 2557889 “Means for treating coccidiosis in birds and animals”, the essence is an anti-eimeriotic pharmaceutical composition containing n-tetradecyltri-n-butylphosphonium bromide and the drug “Erinit” as active components in their weight 1: 2 ratio and glucose as an auxiliary substance.

The well-known drug is used once, the therapeutic dose is 2.5 mg / kg, while the drug has low toxicity.

A disadvantage of the known technical solution is that the known tool is characterized by a rather complex and expensive synthesis of the active substance n-tetradecyltri-n-butylphosphonium bromide, which does not contribute to its widespread use, while neither the drug itself nor its components are available in the Russian Federation, because It is synthesized exclusively in the laboratory conditions of Kazan Federal University, while the invention according to this patent also belongs to KFU and the invention has not been brought to the stage of industrial production also due to lack of investment.

From the studied prior art, the applicant revealed the invention according to RF patent 2629316 "Means for the treatment of nematodoses and eimerioses in veterinary medicine", the essence is a tool for the treatment of nematodes and eimerioses in veterinary medicine, containing n-hexadecyl triphenylphosphonium bromide as an active component and sunflower oil at a weight ratio of 1 : 99. The known drug has low toxicity (LD ₅₀ 225 mg / kg - hazard III), has high therapeutic efficacy at low therapeutic doses.

A disadvantage of the known technical solution is the difficulty of industrial application, since the production of alkylphosphonium salts in Russia is not established and they are not synthesized by the factory method, but were first synthesized at Kazan Federal University (laboratory synthesis), while the invention according to this patent also belongs to KFU and the invention does not brought to the stage of industrial production also due to lack of investment.

The analogues described above are analogues of the claimed technical solution for its intended purpose.

At the same time, attention should be paid to the fact that the prior art has not revealed any analogues in the structure of the active substance N- (4-bromobenzelidene) octadecane-1-amine from the prior art, therefore, the claims are made without a limiting part.

Thus, from the studied prior art, it can be concluded that the search for new drugs and schemes for their use for the prevention and treatment of eimerioses of animals, birds and then humans, acting on various parts of the pathological process, is an urgent task of both veterinary medicine and pharmaceuticals in general .

The objective and technical result of the claimed technical solution is the creation of an effective, non-toxic, new-generation anti-emeriose agent for veterinary medicine with low therapeutic doses, expanding the arsenal of drugs for this purpose, based on N- (4-bromobenzelidene) octadecane-1-amine, which has a characteristic set of properties:

- low therapeutic doses in terms of the active substance - N- (4-bromobenzelidene) octadecane-1-amine;

- lack of toxicity;

- a convenient method of drug administration during animal feeding in the form of an oil solution;

- high efficiency in relation to eimeria, providing an expansion of the arsenal of known products, which at the filing date of the application materials are mostly ineffective, toxic and expensive.

The essence of the claimed technical solution is a tool for the treatment of eimeriosis in animals, containing N- (4-bromobenzelidene) octadecane-1-amine as an active substance and sunflower oil as a solvent in their weight ratio: N- (4-bromobenzelidene) octadecane-1 -amine: sunflower oil = 1: 99.

The claimed technical solution is illustrated in Fig.1-3.

Figure 1 presents Table 1, which shows the comparative effectiveness of the claimed funds based on N- (4-bromobenzelidene) octadecane-1-amine and the well-known drug furazolidone used in the treatment of eimeriosis in cattle (calves), as the most affordable drug.

Figure 2 presents Table 2, which shows the comparative effectiveness of the claimed funds based on N- (4-bromobenzelidene) octadecane-1-amine and the well-known drug amprolium used in the treatment of eimeriosis of small cattle (kids).

Figure 3 presents Table 3, which shows the comparative effectiveness of the claimed funds based on N- (4-bromobenzelidene) octadecane-1-amine and the well-known drug "Baykoks 2.5%" used in the treatment of eimeriosis of rabbits.

Abbreviations in the Tables mean:

- ind. - the number of eggs;

- AI - intensification;

- IE - intensification;

- EE - extensibility.

The tasks and the claimed technical result are achieved through the synthesis of a fundamentally new tool for the treatment of eimeriosis in animals, obtained for the first time in the world, consisting of the active substance N- (4-bromobenzelidene) octadecane-1-amine dissolved in sunflower oil, with a weight ratio of 1 : 99:

N- (4-bromobenzelidene) octadecan-1-amine

The claimed technical solution is as follows.

Example 1. Obtaining the claimed funds based on N- (4-bromobenzelidene) octadecane-1-amine for the treatment of eimeriosis in animals.

The claimed tool is obtained in two stages.

Step 1. Obtaining the active component of the claimed funds is N- (4-bromobenzelidene) octadecane-1-amine.

The active component of the claimed funds is N- (4-bromobenzelidene) octadecane-1-amine of the formula (1) obtained in a known manner described by the applicant in the patent for the invention of the Russian Federation No. 2640816, publ. 01/18/18, Bull. No. 2. Galkina I.V., Bakhtiyarova Yu.V., Gerasimov A.V., Shulaeva M.P., Pozdeev OK, Galkin V.I. "The use of aromatic azomethines with higher alkyl substituents as a means having bactericidal and fungicidal activity", given (method of preparation) by the applicant further.

(one)

To a solution of 2.063 g (0.0112 mol) of 4-bromo-benzaldehyde in 6 ml of warm ethanol, for example, at 30 ° С, a solution of 3 g (0.0112 mol) of octadecylamine-1 in 15 ml of the same solvent (ethanol) is added . The reaction mixture is heated in a water bath at a temperature of 30-50 ° C for, for example, 2 hours, then the reaction mixture is cooled.

The solvent is removed on a rotary evaporator, the precipitate is washed on a Schott funnel with ethanol. N- (4-bromobenzelidene) octadecane-1-amine is obtained in the form of a white powdery product.

Characteristics of the resulting product: so pl. 59.5 ° C. Yield 4.75 g (97.3%). IR spectrum, cm ^-1 : 1644 (C = N). ¹ H NMR (400 MHz, CDCl ₃ ): 8.23 s (1H, -C H = N-), 7.61 d (2H, NC-C = C H -, ³ J _HH = 8.39 Hz), 7.55 d (2H, BrC = C H -CH =, ³ J _HH = 8.40 Hz), 3.61 t. (2H, = NC H ₂ -, ³ J _HH = 6.81 Hz), 1.70 m. (2H, = NCH ₂ -C H ₂ - , ³ J _HH = 13.60 Hz), 1.27 m (30H, [-C H ₂ -] ₁₅ , ³ J _HH = 15.64 Hz), 0.90 t. (3H, -CH ₂ -C H ₃ , ³ J _HH = 7.00 Hz). Calculated,%: C 68.81; H 9.63; N, 3.21. C ₂₅ H ₄₂ NBr. Found,%: C 68.54; H 9.79; N, 3.09.

Stage 2. Obtaining the target product - the claimed funds for the treatment of eimeriosis in animals.

Actually the claimed tool for the treatment of eimeriosis in animals is prepared as follows.

In a porcelain mortar, 1.0 g of N- (4-bromobenzelidene) octadecane-1-amine obtained in Step 1 is triturated until a colorless, uniform powder is obtained and dissolved in 99.0 g of sunflower oil at a temperature of, for example, 35 ° C. Unrefined sunflower oil, first grade, cold pressed, GOST 1129-2013 was used.

For treatment, an oily dosage form is fed to animals during morning feeding. For dosing in animals using prepared 1% solution of N- (4-bromobenzelidene) octadecane-1-amine in sunflower oil in an amount depending on the weight of the animal.

Example 2. The study of the anti-emery efficacy of the claimed funds based on N- (4-bromobenzelidene) octadecane-1-amine.

The anti-emery efficacy of the claimed drug based on N- (4-bromobenzelidene) octadecane-1-amine was studied in three experiments, for example:

- cattle - calves (experiment 1);

- small cattle - kids (experiment 2);

- rabbits (experiment 3).

Experience 1. The study of the anti-emery efficacy of the claimed funds based on N- (4-bromobenzelidene) -octadecane-1-amine in eimeriosis of cattle (calves).

We studied 30 calves of 4-5 months of age, naturally infested with Eimeria bovis, Eimeria zuernii and Eimeria ellipsoidalis contained in Tatnefteproduct LLC of the Atninsky district of the Republic of Tatarstan.

Clinical signs were characterized by emaciation, periodically appearing diarrhea, general depression and loss of appetite.

Based on the results of a coprological study, calves were divided into 3 groups (10 animals each), two of them experimental (1st and 2nd) and one control (3rd). Animals in groups were selected according to the principle of analogues.

Group 1 calves were once given N- (4-bromobenzelidene) octadecane-1-amine at a dose of 10 mg / kg, twice with an interval of 6 days.

Calves of the 2nd group received the well-known drug furazolidone (20 mg / kg) in two five-day courses with an interval of 3 days.

Group 3 calves served as control - without treatment.

Calves of all groups were kept under identical conditions, and their feeding was carried out in accordance with zootechnical standards. A clinical examination of the livestock was performed daily. Fecal samples for the study were taken before drug administration and 7, 15 and 30 days after treatment, respectively.

The results of a study of the anti-eimeriotic efficacy of the claimed agent on calves naturally invaded by eimeriosis are presented in FIG. 1 in Table 1.

From the data presented in Table 1, it can be seen that the intensity of eimeriotic invasion in naturally infected calves ranged from 287 ± 2.5 to 339 ± 1.9.

7 days after treatment:

- in animals of the 1st and 2nd experimental groups there was a decrease in oocysts. The intensity of the drugs in the 1st group was 40.0%, in the 2nd - 30.0%.

15 days after treatment:

- in animals of the 1st and 2nd experimental groups there was a further decrease in Aimeria oocysts. In the 1st group, their number was 95 ± 1.9, in the 2nd - 166 ± 1.5, and the intensities - 64.7% and 47.2%, respectively. During this period, eight animals from the 1st group and six from the 2nd were completely freed from the eimeria.

30 days after treatment:

- in the 1st group, the intensity of eimeriotic invasion decreased to 25 ± 2.2 oocysts, the efficacy was 88.7%, and the extensivity was 90.0%.

- in the 2nd group after treatment with furazolidone, the average number of oocysts in calves during this period was 100 ± 2.6, the intensity was 68.1, and the extensivity was 70.0%.

- in the 3rd group (untreated), the intensity of eimeriotic invasion in calves during the experiment increased to 375 ± 3.2 oocysts, against 339 ± 1.9 at the beginning of the experiment.

From the above it can be concluded that the intensity of the claimed funds based on N- (4-bromobenzelidene) octadecane-1-amine in cattle eimeriosis in the study after 7 days was higher in comparison with furazolidone by 9.0%, after 15 days - by 17.5%, after 30 days - by 20.6%, and ectensivity was higher by 20.0%, respectively.

Experience 2. The study of the anti-emery efficacy of the claimed funds based on N- (4-bromobenzelidene) -octadecane-1-amine in eimeriosis of small cattle (kids).

The study of the antiparasitic effectiveness of the claimed funds on the basis of N- (4-bromobenzelidene) octadecane-1-amine for eimeriosis of small cattle was carried out in the peasant farm (farm) "Abdrakhmanov AA" Vysokogorsky district of the Republic of Tatarstan on 32 goats 3-4 months age, naturally infected with pathogens of eimeriosis.

Before the start of the experiment, fresh samples of feces were selectively taken in buildings 1 and 2, which were placed in test tubes and moistened with a 2% solution of potassium dichromate. The tubes were labeled and delivered to the Department of Epizootology and Parasitology, Federal State Budget Educational Institution of Higher Health Care. Samples were investigated by the Kotelnikov-Khrenov method. To determine the species affiliation of the revealed eimeria, the samples were transferred to bacteriological plates and placed for maturation of oocysts in a thermostat with a temperature of 27-28 ° С for 10 days. During this time, feces were examined daily for oocyst detection. Oocysts that floated to the surface of the flotation liquid were placed on a glass slide, on top of which a coverslip was placed and the species composition of oocysts (vol. × 40, approx. × 10) was studied according to the generally accepted method. In the studied samples, oocysts of the following species were identified: Eimeria faurei, Eimeria intricate, Eimeria ninaekohljakimovae. Based on the results of a coprological study, the animals were divided into 3 groups of 10 -12 animals each.

Before the introduction of the drugs, the kids were kept on a hungry diet for 12 hours, the watering was not limited.

The group 1 goats were orally treated with a bottle of an oil solution of N- (4-bromobenzelidene) octadecane-1-amine at a dose of 10 mg / kg twice with an interval of 6 days.

The goats of the 2nd group in a mixture with concentrates were given the well-known preparation amprolium 30% at a dose of 240 mg of DV per 1 liter of water for 7 days.

The 3rd group of kids was the control and they were not subjected to treatment.

The kids were monitored for 30 days. Fecal samples were taken and examined after 7, 15 and 30 days, respectively, after treatment. The results of the study are presented in FIG. 2 in Table 2.

From the data presented in Table 2, it is seen that:

7 days after treatment:

- in the kids of the 1st group who received the claimed agent on the basis of N- (4-bromobenzelidene) octadecane-1-amine, the II was 59 ± 2.4, the IE and EE of the drug were 45.4% and 30%, respectively.

- in goats of the 2nd group treated with the drug amprolium, these indicators were 71 ± 1.8; 36.5 and 20.0%, respectively.

- in goats of the 3rd, control group, AI increased and amounted to 120 ± 3.1 oocysts in one gram of feces during this period.

15 days after treatment:

- in the kids of the 1st group, AI, IE and EE were 55 ± 1.7; 66.1% and 70%

- in the kids of the 2nd group, AI, IE and EE were 69 ± 1.4; 45% and 50.0%, respectively.

- in the kids of the 3rd, control group, the AI increased to 125 ± 2.5 (Table 2 in Fig. 2).

30 days after treatment:

- in the kids of the 1st group, the indicators of AI, IE and EE were 4 ± 2.3; 86.3% and 80.0%,

- in kids of the 2nd group, the indicators of AI, IE and EE were 21.8 ± 2.4; 69.1% and 60.0%,

- in the goats of the 3rd, control group, the AI increased to 128 ± 3.4 and there was a depressed state, a deterioration in appetite, feces were liquid with an admixture of blood and mucus with an unpleasant odor, the animals mostly lay and they thirsted.

Thus, from the data shown in Table 2, we can conclude that the claimed drug based on N- (4-bromobenzelidene) octadecane-1-amine in goat eimeriosis is more effective than the well-known drug amprolium.

Experience 3. The study of the anti-emery efficacy of the claimed funds based on N- (4-bromobenzelidene) -octadecane-1-amine on the example of rabbits.

In the third experiment, 30 young rabbits of 2 months of age weighing 2-2.5 kg were used, which were kept in the vivarium of the department of epizootology and parasitology of the FSBEI HE KGAVM. Studies were conducted in October - November 2018.

As a result of a coproscopic study of fecal samples by the Kotelnikov-Khrenov method, it was found that the animals are infected with eimeriosis.

The source of invasion was sick rabbits, which were previously kept in this room. Disinvasion has not been performed. This contributed to the natural infection of rabbits in this experiment.

To determine the species affiliation, Eimeria took samples of feces and delivered to the department of epizootology and parasitology, where they studied the species affiliation, using M.V. Krylova.

When considering each type of eimeria, they paid attention to the shape and color of the shells, the presence of residual bodies in the oocyst and spore, and observed the sporogony process. The sizes of oocysts, spores, sporozoites, the thickness of the shells of oocysts and the size of the caps were determined by measuring 150 oocysts of each species using a screw micrometer, which was fixed on the eyepiece of the microscope. The sporulated oocysts were counted in the VIGIS counting chamber.

As a result of studies, it was found that rabbits are infected with a mixed culture of oocysts of the following species: Eimeria stiedae, Eimeria perforans, Eimeria magna.

After this, rabbits were divided into 2 experimental and 1 control groups:

Group 1 rabbits were orally treated with a bottle of an oil solution of N- (4-bromobenzelidene) octadecane-1-amine at a dose of 10 mg / kg twice with an interval of 6 days.

Rabbits of the 2nd group received the well-known preparation “Baykoks 2.5%” at a dose of 7 mg / kg with water for two days in a row.

Group 3 rabbits served as controls; they were not treated.

The rabbits of each group were kept in individual cells during the experiment. The conditions of feeding and feeding were in accordance with the hygiene standards. Fed with dry food for rabbits.

The results of determining the intensity of invasion, intensification and extensibility are presented in Figure 3 in Table 3.

From the data presented in Table 3, it can be seen that prior to treatment, all rabbits were infected with eimeriosis. The invasion intensity ranged from 148 ± 8.4 to 169 ± 9.1 Eimeria oocysts per 1 g of litter.

In group 1 rabbits treated with N- (4-bromobenzelidene) octadecane-1-amine:

- 3 days after the start of treatment, the intensity of invasion decreased significantly and amounted to 28 ± 1.9 aimeria oocysts, while the efficiency was 79.3%, and the extensivity was 80%.

- after 7 days, there was a further decrease in the intensity of invasion - this indicator was 18 ± 1.9 oocysts, the intensity and extensivity were 85.6% and 90%, respectively.

- after 14 days, the rabbit body was completely freed from Eimeria oocysts. The appetite, the general condition of the rabbits, and the consistency of feces during this period returned to normal. No deaths were noted, but some individuals had loose stools. The intensity and extensivity on days 14 and 21 was 100%, which indicates the effective action of N- (4-bromobenzelidene) octadecane-1-amine in rabbit eimeriosis.

In rabbits of the second group receiving the well-known drug Baykoks 2.5%:

- after 3 days, the intensity of invasion was 41 ± 4.8 Aimeria oocysts, the efficacy was 74.4%, and the extensivity was 50%.

- after 7 days, the intensity of invasion decreased and amounted to 26 ± 1.9, while the efficacy and extensibility were 82.6% and 80%, respectively.

- after 14 days and 21 days there was a further decrease in the intensity of eimeriotic invasion. The drug Baykoks also showed high antiparasitic efficacy, but it was lower than when using the claimed funds based on N- (4-bromobenzelidene) octadecane-1-amine. For example, on day 14, Baycox IE amounted to 84.5%, EE - 90.0%, versus 100% when administering the claimed agent based on N- (4-bromobenzelidene) octadecane-1-amine, that is, it was lower by 15, 5% and 10%, respectively.

A similar pattern was observed in studies and on 3, 7 and 21 days after treatment.

In rabbits of the 3rd, control group, the invasion intensity increased from 164 ± 7.1 to 230 ± 8.1 aimeria oocysts in 1 gram of feces. In animals, by the end of the experiment, the mortality rate was 20% of the total population, clinical signs of eimeriosis were observed: lethargy, weight loss, liquid feces mixed with blood, decreased appetite (the food remained in the feeder for the morning), anemia of the mucous membranes of the eyes, nasal and oral cavity .

Thus, the analysis of the data presented in Table 3 in Figure 3 shows that the claimed drug based on N- (4-bromobenzelidene) octadecane-1-amine at a dose of 10 mg / kg when administered twice with an interval of 6 days completely free the body rabbits from eimeria 14 days after treatment and has a higher anti-eimeriotic efficacy.

Example 3. Determination of toxicity of the claimed funds based on N- (4-bromobenzelidene) octadecane-1-amine.

The toxicity of the claimed N- (4-bromobenzelidene) octadecan-1-amine-based agent was studied in non-linear white mice. The work was carried out at the Department of Epizootology and Parasitology, FSBEI HE KGAVM in the period from December 2018 to January 2019.

The toxic effect was evaluated in accordance with the guidelines for determining the toxic properties of drugs used in veterinary medicine and animal husbandry. Observation of the mice was carried out for 14 days from the time of administration of the drug. At the same time, the appearance and behavior of mice, the condition of the coat and visible mucous membranes, the attitude to the feed, the mobility, the time of occurrence and the nature of intoxication, its severity, reversibility, timing of death, the number of dead and surviving animals were taken into account. Deaths were recorded and an autopsy was performed.

During the experiment, the mice were in the same conditions. Feeding was carried out in accordance with veterinary and zootechnical standards. A clinical examination of the livestock was performed daily.

In the experiment, 42 white mice, of both sexes, weighing 18-22 g. Were used. Laboratory mice, taking into account gender, age, body weight, were divided into 7 groups, 6 experimental and one control (6 animals each). The drug compound was dissolved in warm water, injected once into the stomach in various doses with a club-shaped needle.

The mice of the six experimental groups were given 400 mg / kg, 600 mg / kg, 800 mg / kg, 1000 mg / kg, 3000 mg / kg and 5000 mg / kg. Control mice were injected with 0.5 ml of water. Mice were allowed to feed 2 hours after drug administration. Access to water was not limited.

The clinical condition of the mice of the experimental groups did not differ from that in the mice of the control group. Mice willingly ate food, adequately responded to tactile, pain, sound and light stimuli. The condition of the skin and mucous membranes of the mice of the experimental groups was the same as that of the mice of the control group.

During the experiment, an autopsy was performed on mice. An autopsy was performed on one animal from each group on days 3 and 7 of the experiment. Apparent changes in the organs of the chest and abdominal cavities in mice of both the control and experimental groups were not found by the applicant.

In this case, the average lethal dose (LD50) was not calculated due to the lack of death of animals in all 6 experimental groups. The maximum dose of the claimed funds administered to mice of the 6th group was 5000 mg / kg

Thus, the claimed agent based on N- (4-bromobenzelidene) octadecane-1-amine at a dose of 5000 mg / kg does not have a toxic effect and belongs to hazard class IV.

Example 4. The study of the irritating effect of the claimed funds based on N- (4-bromobenzelidene) octadecane-1-amine on the mucous membranes of the eyes

The irritating effect of the claimed agent based on N- (4-bromobenzelidene) octadecane-1-amine on the mucous membranes of the eyes was studied in 15 rabbits weighing 2.5-3 kg. Animals of the experimental groups on the conjunctiva of the right eye using an eye pipette once instilled 2 drops of 0.1%, 0.3%, 0.5%, 0.7% and 1% aqueous solution of the claimed funds based on N- (4-bromobenzelide ) octadecan-1-amine, and the left - water at room temperature (18-20 ° C). When applying the claimed funds, the upper corner of the conjunctival sac was pulled, then the lacrimal-nasal canal was pressed for 1 minute. The condition of the animals was evaluated after 5, 30, 60 minutes after applying the claimed funds and daily for 14 days. Pay attention to the condition of the lining of the eye, tearing, swelling, hyperemia.

The reaction of the mucous membrane of the eye was scored on a five-point scale.

We monitored the appearance of hyperemia, swelling, injection of scleral vessels and the condition of the pupils. The effect of the solution was also evaluated by the nature of conjunctivitis (superficial, deep), burns of the mucosa of the I, II, III degrees.

Studies have shown that:

- solutions of the claimed funds based on N- (4-bromobenzelidene) octadecane-1-amine in 0.1-0.5% concentration during the study period do not adversely affect the mucous membranes.

- after the administration of a 0.7% solution of the claimed drug, slight lacrimation was noted, which spontaneously stopped 20-30 minutes after the drug was administered.

- after the introduction of the claimed funds in 1% concentration, the rabbits tried to scratch the eye, sat with narrowed eyes, after 2 hours the mucous membrane of the eyes became hyperemic, there was slight swelling, lacrimation. The next day, lacrimation ceased, swelling subsided, the color of the mucous membranes was pink, on the third day it turned pale pink.

Thus, from the foregoing, it can be concluded that the applicant has achieved the stated objectives and the claimed technical results , namely, he created an effective, non-toxic, new generation anti-eimeriotic agent for veterinary medicine with low therapeutic doses, expanding the arsenal of drugs for this purpose, based on N- ( 4-bromobenzelidene) octadecan-1-amine, which has a characteristic set of properties:

- low therapeutic doses in terms of the active substance - N- (4-bromobenzelidene) octadecane-1-amine - the drug is taken orally in the form of an oil solution during morning feeding - once at a dose of 10 mg / kg of animal’s live weight and a second time in the same dose after 6 days, i.e. a total of 20 mg / kg for the entire course of treatment;

- lack of toxicity - LD50 2500 mg / kg (hazard class IV);

- a convenient method of drug administration during animal feeding in the form of an oil solution;

- high efficiency against eymeria, providing an expansion of the arsenal of known ineffective, toxic and expensive drugs for this purpose.

So, for example, with eimeriosis of cattle (calves), the intensity of the claimed agent based on N- (4-bromobenzelidene) octadecane-1-amine in the study after 7 days was higher in comparison with furazolidone by 9.0%, after 15 days - by 17.5%, after 30 days - by 20.6%, and ectensivity was higher by 20.0%, respectively.

The claimed technical solution meets the criterion of "novelty" presented to the invention, since the investigated prior art has not identified technical solutions that have the claimed combination of features that achieve the stated results.

The claimed technical solution meets the criterion of "inventive step" for inventions. A new structure-created compound N- (4-bromobenzelidene) octadecane-1-amine, not known in the world at the date of submission of application materials, is an active component of the claimed anti-emeriosis agent, which is not obvious to a person skilled in the art, due to which the formula The invention is made without a restrictive part.

The claimed technical solution meets the criterion of "industrial applicability" to the invention, because The claimed drug can be obtained through the use of known components using standard equipment and known techniques.

Claims (3)

A drug for the treatment of eimeriosis in animals, containing N- (4-bromobenzelidene) octadecane-1-amine as an active substance and sunflower oil as a solvent in their weight ratio:  N- (4-bromobenzelidene) octadecane-1-amine: sunflower oil = 1: 99.

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