RU2532355C1 - Method of treating poultry with polycationic triviron compound - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves administering into the poultry the polycationic compound 1,5-bis-(4-tetradecyl-1,4-diazoniabicyclo[2.2.2]octan-1-yl)pentan tetrabromide dissolved in 85% formic acid with reducing the concentration to 3 gram per 1 litre of the acid; then the mother solution is diluted with tap water to the concentration of the mother solution of 10 ml per 1 litre of water, and administered orally into the poultry.

EFFECT: invention provides the antiviral and antibacterial action, higher efficacy of treating and preventing the poultry's viral infections caused by viral RNAs, a possibility to administer the therapeutic solution orally through a watering system.

9 cl, 1 tbl, 8 ex

Description

The invention relates to the field of veterinary virology and is intended for the treatment of viral diseases in poultry based on RNA viruses (influenza viruses, acute respiratory infection, hepatitis, blue comb, infectious enteritis, vector-borne enteritis, reovirus enteronephritis, tenosynovitis, viral arthritis, infectious chicken bronchitis, Gamboro disease, pneumovirus, etc.).

Currently, a number of antiviral drugs are known (they belong to different classes of chemical compounds) recommended for use in medical practice. However, due to their high cost, only some of them have been tested as therapeutic agents for viral diseases, as a rule in the treatment of expensive dog and cat breeds. In some cases, when there are no drugs specific for specific viral infections, immunomodulators are used to treat viral diseases. However, such drugs used in medical practice also have a high cost, which excludes the possibility of their widespread use in veterinary practice.

Compounds capable of cleaving RNAs (artificial RNases) have antiviral activity against RNA containing viruses (Goncharova, E.P. et. Al. Inactivation of the tick-borne encephalitis virus by RNA-cleaving compounds // J. Mol. Gen. Med., 2011, V. 5, P. 266-272; Fedorova AA et.al. Inactivation of a non-enveloped RNA virus by artificial ribonucleases: Honey bees and acute bee paralysis virus as a new experimental model for in vivo antiviral activity assessment // Antiviral Res. 2011 V. 91, P. 267-277; RU 2399388, 09.20.2010). In particular, artificial RNases based on derivatives of 1,4-diazabicyclo [2.2.2] octane, namely 1,5-bis- (4-dodecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane dichromide dibromide, have antiviral activity against influenza viruses A / WSN / 33 (H1N1) and tick-borne encephalitis virus (Sofin strain, RU 2399669, 09/20/2010). However, the concentration of this compound necessary for the inactivation of these viruses was 20 μM, which was almost two times the concentration at which toxic effects were observed on cell culture (12 μM).

Some other derivatives of the bis-quaternary salts of 1,4-diazabicyclo [2.2.2] octane, being covalently attached to various surfaces, give the materials thus obtained antiviral properties.

Known drug Argovit (RU 2147237, 10.04.2000). This drug has a wide spectrum of antimicrobial activity against gram-positive and gram-negative bacteria. It exhibits virucidal (antiviral) and fungicidal (antifungal) activity. The disadvantages, in addition to the high cost, include the fact that the drug is available in the form of a stabilized concentrated 20% solution (the silver content in argovite in terms of 100% dry preparation is 5-7%). The form of release of the drug imposes significant restrictions on the conditions and shelf life of the drug. So, during long-term storage and especially under the influence of factors such as light, temperature fluctuations, freezing, aggregation of cluster particles occurs with the formation of colloidal, inactive silver particles. A silver metal film may also appear on the surface of the concentrate. Silver particles are not absorbed, which also limits the treatment of systemic and respiratory viral infections of birds by drinking the drug.

The closest analogue of the claimed invention is a method for the prevention and treatment of diseases of birds of bacterial and viral etiology, in which a sick bird is instilled in the beak 1-7 grains in dissolved form a complex homeopathic remedy containing Lachesis, Echinacea, Aconitum, Bryonia, Belladonna, taken equally, after 0.5-6 hours for 1-6 weeks (RU 2209076, 07.27.2003).

The technical result of the present invention is to expand the arsenal of antiviral agents for veterinary medicine, increase the effectiveness of the treatment of viral diseases in poultry, which are based on RNA viruses, and provide the possibility of prevention and treatment of these diseases, after they have already arisen, as well as the ability to do so using input through the drinking system orally, and not by injection intramuscularly.

The specified technical result is achieved in a method of treating viral diseases of birds caused by RNA viruses, in which the polycationic compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) is introduced into the body of birds. pentane tetrabromide, which is dissolved in formic 85% acid, bringing the concentration to 3 grams per 1 liter of acid, obtaining a mother liquor, then the mother liquor is diluted with tap water to a concentration of 10 ml of the mother liquor per 1 liter of water and administered orally to the bird.

The compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0.00015 ml of mother liquor per 1 bird's head in the first 3 days and 0, 0003 ml of the mother liquor over the next two days.

The compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0.0003 ml of mother liquor per 1 bird head in the first 5 days and 0, 00045 ml of the mother liquor per bird head over the next two days.

The compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0.00045 ml of mother liquor per bird head in the first 5 days and 0.0009 ml of mother liquor over the next three days.

The compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0.00045 ml of mother liquor per bird head in the first 5 days and 0.001 ml in over the next two days.

The compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0.00045 ml of mother liquor per bird head in the first 5 days and 0.0009 ml over the next five days.

The compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0.00025 ml of mother liquor per bird head in the first 5 days and 0.001 ml in as a mother liquor over the next three days.

The compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0.00025 ml of mother liquor per head for the first 5 days and 0.001 ml for the next two days.

The compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0.00025 ml of mother liquor per bird head in the first 6 days and 0.001 ml in for the next three days.

Administration of the preparation 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is carried out as follows.

First, the mother liquor is prepared by dissolving the drug in formic 85% acid, bringing the concentration to 3 grams of the drug per 1 liter of acid, then the mother liquor is diluted with tap water to a concentration of 10 ml of the mother liquor per 1 liter of water. Diluted with water, the mother liquor is introduced into the watering system, ensuring the dosage of the mother liquor = 0.00015-0.001 ml per head per day for 5-10 days.

The compound in undissolved form is stored and retains its properties for 3 years.

Unlike analogues, it is administered orally, which makes it possible to use it in industrial poultry farming for the treatment of viral diseases of birds.

The polycationic compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide (C ₄₇ H ₁₀₀ Br ₄ N ₄ ) belongs to the class of polycation saturated heterocyclic compounds containing hydrophobic alkyl fragments, and is a derivative of 1,4-diazabicyclo [2.2.2] octane and has the following structural formula:

Below is the synthesis scheme for 1,5-bis- (4-dodecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide (I) (hereinafter, the preparation is dry or dissolved in 85% formic acid with the name "TRIVIRON"):

The method of synthesis / preparation of a derivative of 1,4-diazabicyclo [2.2.2] octane with the structural formula

the synthesis is carried out at 55 ° C for 24 hours and a molar ratio of the following components: 1,5-dibromopentane - 1-tetradecyl-4-aza-1-azoniabicyclo [2.2.2] octane 1: 2.16 bromide , in methanol as a solvent, with a yield of the target compound exceeding 89%.

An example of obtaining the preparation "TRIVIRON" (polycationic compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide (I)).

1-Tetradecyl-4-aza-1-azoniabicyclo [2.2.2] octane bromide (26.3 g, 65 mmol) is dissolved by heating to 55 ° C in 20 ml of methanol. To the solution, with stirring on a magnetic stirrer, 1.5-dibromopentane (6.9 g, 30 mmol) are added in two portions at intervals of 12 hours. After the last portion was added, the reaction mixture was stirred for 24 hours at 55 ° C. After cooling to room temperature (18-22 ° C), 20 ml of acetonitrile was added to the reaction mixture, the precipitate formed was filtered off and dried in vacuo (17.5 g, 56%). The mother liquor is evaporated on a rotary evaporator, 40 ml of acetonitrile is added to the viscous syrupy residue, stirred on a magnetic stirrer at the boil under reflux for 1 hour. After cooling to room temperature (18-22 ° C), the precipitate formed is filtered off and dried in vacuum (10, 3 g, 33%). The resulting precipitation is combined. Yield 27.8 g (89.1%).

¹ H NMR spectrum, δ, ppm (J, Hz): 0.93 (br t, 6H, C H ₃ (CH ₂ ) ₁₁ N ⁺ , J 6.6), 1.32-1.45 (m, 36H, CH ₃ (C H ₂ ) ₉ CH ₂ CH ₂ N ⁺ ); 1.32 (m, 2H, ⁺ NCH ₂ CH ₂ C H ₂ CH ₂ CH2N ⁺ ); 1.88 (m, 4H, CH ₃ (CH ₂ ) ₉ C H ₂ CH ₂ N ⁺ ); 1.98 (m, 4H, ⁺ NCH ₂ C H ₂ CH ₂ C H ₂ CH ₂ N ⁺ ); 3.62 (m, 4H, CH ₃ (CH ₂ ) ₁₀ C H ₂ N ⁺ ); 3.75 (m, 4H, CH ₂ C H ₂ N ⁺ ); 4.07-4.12 (m, 24H, ⁺ N (C H ₂ C H ₂ ) ₃ N ⁺ ).

¹³ C NMR spectrum, δ, ppm: 12.95 (2C, C H ₃ (CH ₂ ) ₁₁ N ⁺ ); 21.64 (2C, CH ₃ C H ₂ (CH ₂ ) ₁₀ N ⁺ ); 22.21 (2C, CH ₃ CH ₂ C H ₂ (CH ₂ ) ₉ N ⁺ ); 25.63 (4C, ⁺ NCH ₂ C H ₂ CH ₂ C H ₂ CH ₂ N ⁺ , CH ₃ (CH ₂ ) ₂ C H ₂ (CH ₂ ) ₈ N ⁺ ); 28.66 (2C, CH ₃ (CH ₂ ) ₃ C H ₂ (CH ₂ ) ₇ N ⁺ ); 28.95 (4C, CH ₃ (CH ₂ ) ₄ C H ₂ C H ₂ (CH ₂ ) ₅ N ⁺ ); 29.10 (3C, ⁺ N (CH ₂ ) ₂ C H ₂ (CH ₂ ) ₂ N ⁺ , CH ₃ (CH ₂ ) ₆ C H ₂ CH ₂ (CH ₂ ) ₃ N ⁺ ); 29.21 (2C, CH ₃ (CH ₂ ) ₈ C H ₂ (CH ₂ ) ₂ N ⁺ ); 31.54 (2C, CH ₃ (CH ₂ ) - C H ₂ CH ₂ N ⁺ ); 64.24 (2C, CH ₃ (CH ₂ ) ₁₀ C H ₂ N ⁺ ); 50.81 and 51.03 (6C, ⁺ N ( C H ₂ C H ₂ ) ₃ N ⁺ ); 64.71 (2C, C H ₂ ⁺ N (CH ₂ CH ₂ ) ₃ N ⁺ ).

The drug "TRIVIRON" is used for the prevention and / or treatment of viral diseases of poultry, which are based on RNA viruses, this drug is administered orally, which can significantly improve the health of the bird, reduce its mortality, eliminate delay in weight gain, etc.

Examples of evaluating the antiviral activity of the drug "TRIVIRON" when tested on poultry.

Example No. 1. The use of the drug "TRIVIRON" in an industrial herd in order to reduce mortality in the number of broilers affected by infectious Newcastle disease.

Mortality at the poultry farm was from 28 days - 0.7% per day, the bird was inhibited, the feed did not consume, the litter was liquid.

At autopsy, the pathological signs were: hemorrhagic inflammation of the trachea, aerosacculitis, enterocolitis, hemorrhagic proventriculitis.

The use of the drug "TRIVIRON" began on a population of 30,000 animals at the age of 31 days. The drug was drunk around the clock to ensure consumption by the entire herd of the drug for 5 days. First, a mother liquor was prepared by dissolving in formic acid to a concentration of 3 grams of the drug per 1 liter of 85% formic acid, then the dissolved compound was diluted with tap water to a concentration of 10 ml of mother liquor per 1 liter of water, the drug was introduced into the watering system, providing the amount of mother liquor a solution of the drug 0.00015 ml per 1 head in the first 3 days and 0.0003 ml of the mother liquor over the next two days.

Mortality / mortality decreased after three days of use of the drug to 0.3% per day, i.e. more than 2 times, the bird began to consume food.

A postmortem examination of a dead bird revealed a decrease in the number of fibrinous deposits in the chest and abdominal cavities.

After five days of treatment, mortality decreased to 0.2% per day.

In the experimental group, seroconversion was not observed for the Newcastle disease virus, i.e. antibody growth in response to exposure to the virus.

Example No. 2. was carried out at the enterprise with the productive parental livestock of the Hubbard cross affected by a variant strain of Infectious bird brochitis (the diagnosis was confirmed by virological studies). The drug was used on a stock of birds at the age of 50 weeks for 7 days with water. First, the mother liquor was prepared by dissolving in formic acid to a concentration of 3 grams of the drug per 1 liter of 85% formic acid, then the dissolved compound was diluted with tap water to a concentration of 10 ml of the mother liquor per 1 liter of water, the drug was introduced into the watering system, providing a dose 0.0003 ml of the mother liquor per head of the drug in the first 5 days and 0.0045 ml of the mother liquor per head for the next two days

During pathological observation, the following changes were revealed - a decrease in the influence of the nephropathogenic virus, i.e. deposition of urates (uric acid salts) in the ureters, the total progressive amount of false laying hens also decreased, the existing cysts on the ovaries remained, and the productive period of the herd lasted without a natural (in age) decrease to 63 weeks. The number of fibrinous deposits in the chest and abdominal cavities of a bird was detected less frequently than before treatment. The number of deformed eggs and unfertilized also decreased. The liver condition also improved - hepatoses and cholecystitis were detected less often than before treatment.

Example No. 3. The purpose of the application was to use the drug "TRIVIRON" as a therapeutic agent against viral pneumovirus infection (ART): pneumovirus was one of the leading causes of poultry death in the pre-slaughter period. The diagnosis made at the factory at autopsy was confirmed by laboratory tests of the selected material.

First, the mother liquor was prepared by dissolving in formic acid to a concentration of 3 grams of the drug per 1 liter of 85% formic acid, then, the dissolved compound was diluted with tap water to a concentration of 10 ml of the mother liquor per 1 liter of water, the drug was introduced into the watering system at a dose of 0 , 00045 ml of the mother liquor per head of the drug in the first 5 days and 0,0009 ml of the mother liquor for the next 3 days.

The result of the use of the drug was a decrease in the case at the time of the peak departure of the bird at the age starting from the 24th day of rearing, as well as an improvement in the safety indicators of the bird as a whole, from the 14th day of rearing to slaughter. In parallel, broiler productivity indicators were monitored: safety and average daily gain.

In closed batches, there was a positive tendency to increase the safety, average daily gain and feed conversion in the groups where the TRIVIRON preparation was evaporated, especially where it was evaporated starting from 22-24 days, namely:

- in comparison with the control houses, the average safety index in the houses where TRIVIRON was used was 2.31% higher;

- in comparison with the control houses, the average daily gain in the houses where the TRIVIRON preparation was used was 1.44 grams higher.

The clinical trials of the drug "TRIVIRON" in poultry farms unsuccessful for viral infections caused by RNA viruses are shown in table 1. As a result, we noted the result of the introduction of the drug "TRIVIRON" - an increase in the percentage of safety in closed lots (table 1).

Table 1 No.
an example RNA infectious disease Preservation in the experimental group with the drug "TRIVIRON" Safety in the control group without the drug "TRIVIRON" Dosage of the mother liquor Example No. 4 Chickens Infectious Bronchitis 95% 91.1% 0.00045 ml per head in the first 5 days and 0.001 ml in the second 2 days. Example No. 5 Gumboro's disease 94.7% 89.2% 0.00045 ml in the first 5 days and 0.0009 ml in the second 5 days Example No. 6 Pneumovirus 93.8% 89.1% 0.00025 ml per head in the first 5 days and 0.001 ml in the second 3 days Example No. 7 Reovirus 96.1 90.7% 0.00025 ml per head in the first 5 days and 0.001 ml in the second 2 days. Example No. 8 Pneumovirus 97% 92.1% 0.00025 ml per head in the first 6 days and 0.001 ml in the second 3 days

Thus, the drug "TRIVIRON" has a pronounced antiviral effect against viral diseases of birds, which are based on RNA viruses.

Using the drug "TRIVIRON" allows you to cope with a wide range of infectious diseases, which are based on RNA viruses. Its effectiveness is confirmed by a large number of experiments in factories. A distinctive feature of it is the ability not to prevent and treat these diseases after they have already arisen, as well as the ability to do this using input through the drinking system orally, and not by injection intramuscularly. An advantage is also the absence of side effects.

Claims (9)

1. A method for the treatment of viral diseases of birds caused by RNA viruses, characterized in that the polycationic compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane is introduced into the body of birds tetrabromide, which is dissolved in formic 85% acid, bringing the concentration to 3 grams per 1 liter of acid, obtaining a mother liquor, then the mother liquor is diluted with tap water to a concentration of 10 ml of the mother liquor per 1 liter of water and administered orally to the bird. 2. The method according to claim 1, characterized in that the compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0.00015 ml of mother solution per 1 head of birds in the first 3 days and 0.0003 ml of the mother liquor over the next two days. 3. The method according to claim 1, characterized in that the compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0.0003 ml of mother solution per 1 bird head in the first 5 days and 0.00045 ml of stock solution per bird head over the next two days. 4. The method according to claim 1, characterized in that the compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0,00045 ml of mother solution on the birds head in the first 5 days and 0.0009 ml of the mother liquor over the next three days. 5. The method according to claim 1, characterized in that the compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0,00045 ml of mother solution on the head of birds in the first 5 days and 0.001 ml over the next two days. 6. The method according to claim 1, characterized in that the compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0.00045 ml of mother solution on the birds head in the first 5 days and 0.0009 ml over the next five days. 7. The method according to claim 1, characterized in that the compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0,00025 ml of mother solution on the head of birds in the first 5 days and 0.001 ml in the form of a mother liquor over the next three days. 8. The method according to claim 1, characterized in that the compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0,00025 ml of mother solution per head in the first 5 days and 0.001 ml over the next two days. 9. The method according to claim 1, characterized in that the compound 1,5-bis- (4-tetradecyl-1,4-diazoniabicyclo [2.2.2] octan-1-yl) pentane tetrabromide is administered at the rate of 0,00025 ml of mother solution on the head of birds in the first 6 days and 0.001 ml over the next three days.