RU2751876C1 - Drug for the prevention and treatment of vaginitis in cows - Google Patents

Abstract

FIELD: pharmaceutical industry; veterinary medicine.

SUBSTANCE: invention relates to the field of pharmaceuticals and veterinary medicine, namely to a drug for the prevention and treatment of vaginitis in cows. The drug contains chitosan hydrochloride as an active part with specific molecular weight and ascorbic or boric acid. The drug also contains an auxiliary part, which is a gel-forming base selected from a triblock copolymer of ethylene oxide and propylene oxide or a mixture of gelatin with glycerin, or a triblock copolymer of ethylene oxide and propylene oxide with gelatin, or a triblock copolymer of ethylene oxide and propylene oxide with gelatin and silicon tetraglycerolate, or confectionery fat with beeswax and nonionic surfactant TWEEN-80. The drug also contains water at a certain ratio of components.

EFFECT: invention provides a highly effective biologically active agent based on chitosan hydrochloride for the treatment and prevention of vaginitis in cattle, which does not contain antibiotics and antiseptics, made in the form of an injectable composition or suppository, and the injectable composition at a temperature of less than 4-15°C is a liquid, and at temperatures close to physiological, it undergoes a phase transition into a form-stable hydrogel.

1 cl, 5 tbl, 18 ex, 3 dwg

Description

The invention relates to pharmacology and veterinary medicine, in particular - gynecology, and can be used for the prevention and treatment of vaginitis in cows.

Vaginitis is an inflammation of the vagina that occurs due to the penetration of microorganisms Streptococcus vaginitis into the genitals of an animal. Currently, the main commercially produced veterinary drugs for the treatment of vaginitis contain disinfecting components (furacilin, iodine), antiseptics (potassium permanganate, ichtammol, chlorhexidine hydrochloride) or antibiotics (enrofloxacin, tetracycline, etc.). The presence of antiseptics and antibiotics in the composition of preparations, as a rule, leads to a decrease in the immunity of animals, allergic reactions and other negative consequences. In addition, the widespread use of antibiotics leads to antibiotic resistance of a living organism, necessitating the development and introduction of new dosage forms.

Known drug "Metrolek" in the form of a foam emulsion for the treatment and prevention of postpartum complications, including vaginitis, in cows (see instructions for use of "Metrolek" registration certificate No. PVR-1-35.13 / 02987), containing 0.5% tylosin tartrate in as an active ingredient, 3.3% of excipients (sunflower oil, T-2 emulsifier, beta-carotene) and water for injection.

The disadvantage of the drug is the presence in the composition of the antibiotic - tylosin tartrate, which can initiate the transition of the disease into a latent form without clinical manifestations, and also significantly limits the use of milk for food purposes during and in the first few days after treatment.

As an alternative to antibiotics, it is proposed to use drugs of natural origin with their own antibacterial activity, which can be used as independent veterinary drugs or in the form of a biologically active matrix for prolonged delivery of drugs.

Known agent in the form of an emulsion for the prevention and treatment of gynecological diseases in animals (see RF patent No. 2384323, IPC A61K 9/107, publ. 03/20/2010), containing 20-60% sapropel composition, 5-15% emulsifier OP-10 to stabilize the emulsion and water. Sapropel is a sediment that accumulates on the bottom of fresh water bodies, enriched with biologically active minerals (calcium, phosphorus, sulfur, iron) and trace elements (iodine, cobalt, copper, manganese, bromine).

The disadvantage of the agent should be considered the high content of sapropel in the composition, which is necessary to achieve a therapeutic effect. In small quantities, sapropel has a weak antibacterial and keratolytic effect. The drug is not convenient to use, since, according to the invention, the agent is a concentrate and in order to obtain the required therapeutic concentration for each specific case, it must be pre-diluted.

The natural aminopolysaccharide chitosan can be considered promising for the creation of biologically active drugs. Chitosan in different morphological forms exhibits a wide range of biological activity, and materials based on it have an accelerated wound healing and high anti-inflammatory effect (see RF patent No. 2617501, IPC S08J 3/075, publ. 04/25/2017).

Known biological product "Biometrin" for the prevention and treatment of endometritis in cows, containing a 0.5-1.5% solution of chitosan in 2% acetic acid, a solution of cultures Bacillus subtilis CH 201 and Bacillus licheniformis CH 200 with a spore titer of 10 ⁸ -10 ¹¹ in the ratio 1: 1 and excipients: lactose, CaCO ₃ , whey or beet juice (see RF patent No. 2687041, IPC C12N 1/20, publ. 06.05.2019).

The disadvantage of the presented drug should be considered the use of acetic acid to dissolve chitosan, which can cause irritation of the mucous membrane. According to the invention, a freshly prepared aqueous acidic solution of chitosan is autoclaved at a temperature of 121 ° C, which can lead to destruction and a decrease in the pharmacological activity of chitosan. In this case, a significant decrease in the dynamic viscosity of the initial chitosan solution is also observed. In addition, the use of this biological product is limited to the prevention and treatment of only postpartum endometritis in cows. Treatment of vaginitis in cows was not included in the invention.

The closest technical solution to the claimed invention is an intrauterine device for the treatment and prevention of vulvovaginitis in cows in the form of suppositories (see RF patent No. 2375067, IPC A61K 35/12, publ. 10.12.2009). The candles contain in their composition the natural pharmacopoeial drug "Vivaton", the antiseptic-stimulant drug Dorogov 2nd fraction, consisting of at least 10% of biologically active substances, the emulsifier lanolin, as well as the forming base in the form of cocoa butter, with the following ratio of components to one candle, g: 1.0: 1.0: 0.5: 8.0, respectively. "Vivaton" is a complex of extracts and infusions of medicinal plants and contains 207 components, such as: organic acids and amino acids, including essential ones (lauric, proline, lemon, oily, oxalic, ascorbic, glutamic, folic, succinic, linolenic, apple, etc.), a complex of vitamins (B ₁ , B ₂ , E, C, B ₆ , A, P), choline, provitamins, phytoncides, glycosides, saponins, flavonoids, carotenoids, hatinin, lecithin, catechins, sucrose, terpenoids , tannins, essential oils, macro- and microelements and other components of natural origin (registration certificate No. 98/183/8 dated 4.06.98, TU 112-84-803-3615-001-13). Antiseptic-stimulant Dorogov 2nd fraction - a preparation obtained from livestock products (meat, bones, etc.) by dry sublimation at elevated temperatures (registration certificate No. PVR-3-1.1 / 00937 of 23.08.13). The drug consists of lower carboxylic acids, their ammonium salts, amides and choline esters, various derivatives of ammonia and ammonium salts, choline, methyl mercaptan, compounds with an active sulfhydryl group and water.

The disadvantage is the multicomponent and laboriousness of the manufacturing process of the initial ingredients of the drug, the high content of the emulsifier in the finished product, as well as the dosage in the use of candles - 2 pcs. / 2 times a day for 3-5 days, which is economically costly when used in large farms with high livestock population.

The technical problem of the invention is the creation of a highly effective biologically active agent for the treatment and prevention of vaginitis in cattle, which does not contain antibiotics and antiseptics.

The technical result is to obtain an agent based on chitosan hydrochloride, made in the form of an injectable composition or suppository suppository. An injectable composition should be understood as a composition that at a temperature of less than 4-15 ° C is a liquid, and at temperatures close to physiological (~ 35-37 ° C) undergoes a phase transition into a form-stable hydrogel.

The specified technical result is achieved in that the agent for the prevention and treatment of vaginitis in cows, containing the active and auxiliary parts, according to the invention, contains chitosan hydrochloride as an active part, and a gel-forming base as an auxiliary part, with the following ratio of components, wt% :

chitosan hydrochloride 0.3 - 3.0 gelling base 12.5 - 77.0 water rest

As a gel-forming base, the agent may contain a triblock copolymer of ethylene oxide and propylene oxide in the following ratio of components, wt. %:

chitosan hydrochloride 0.5 - 1.6 triblock copolymer of ethylene oxide and propylene oxide 12.5 - 18.0 water rest

The active part of the agent may additionally contain ascorbic or boric acid in the following ratio of components, wt. %:

chitosan hydrochloride 0.3 - 3.0

ascorbic or boric acid 0.3 - 3.0

gelling base 12.5 - 77.0 water rest

If there is ascorbic or boric acid in the active part of the agent, the agent may contain, as a gel-forming base, a triblock copolymer of ethylene oxide and propylene oxide in the following ratio of components, wt. %:

chitosan hydrochloride 0.9

ascorbic or boric acid 0.9

triblock copolymer of ethylene oxide and propylene oxide 17.5 water rest

The tool may contain in the active part only ascorbic acid, and as a gel-forming base, a mixture of gelatin with glycerin in the following ratio of components, wt. %:

chitosan hydrochloride 3.0 ascorbic acid 3.0 mixture of gelatin with glycerin 40.0 water rest

If only ascorbic acid is present in the active part, the agent may contain, as a gel-forming base, a mixture of a triblock copolymer of ethylene oxide and propylene oxide with gelatin in the following ratio of components, wt. %:

chitosan hydrochloride 0.3 ascorbic acid 0.3 triblock copolymer of ethylene oxide and propylene oxide with gelatin 14.5 water rest

If only ascorbic acid is present in the active part, the agent may contain, as a gel-forming base, a mixture of a triblock copolymer of ethylene oxide and propylene oxide with gelatin and silicon tetraglycerolate in the following ratio of components, wt. %:

chitosan hydrochloride 0.3 ascorbic acid 0.3 triblock copolymer of ethylene oxide and propylene oxide with gelatin and silicon tetraglycerolate 29.0 water rest

If only ascorbic acid is present in the active part, the agent may contain a mixture of confectionery fat and beeswax as a gel-forming base, and additionally contain a non-ionic surfactant TWIN-80 in the following ratio of components, wt. %:

chitosan hydrochloride 0.6 ascorbic acid 0.6 confectionery fat with beeswax 77.0 nonionic surfactant TWIN-80 3.0

Chitosan hydrochloride is used with a molecular weight (MW) of 30-70 kDa and a degree of deacetylation of ≥75-80 mol%.

D- ascorbic acid is an antioxidant, in the composition of veterinary drugs with intravaginal use, it lowers the pH, inhibiting the growth of bacteria, and also helps to restore and maintain normal pH and flora.

Boric acid is a monobasic acid with fungistatic and bacteriostatic activity. It is used in the treatment of fungal diseases.

Pluronic F - 127 (Pl F - 127) is a triblock copolymer of ethylene oxide and propylene oxide with a molecular weight of 12.6 kDa. It is a non-toxic non-ionic surfactant. Approved for use in pharmaceuticals as a base for hydrogels (European Pharmacopoeia 5th Edition, Order Code P2164030).

Silicon tetraglycerolate (Si (OGly) ₄ ) is a solution of silicon glycerolate in glycerol (GlyOH) at a molar ratio of Si (OGly) ₄ : GlyOH = 1: 3. Biologically active substance, exhibits transcutaneous and antibacterial activity. In an aqueous medium, it hydrolyzes with the occurrence of a sol-gel reaction and the formation of a polysiloxane spatial network of Si-О-Si≡ bonds, the nucleation centers of which are free hydroxy groups. Sol-gel hydrogels based on silicon tetraglycerolate are recommended for practical use as independent drugs or bases for pharmaceutical compositions.

Glycerin (Gly (OH) ₃ ) in the water-glycerol medium is a solvent for silicon tetraglycerolate and a plasticizer for chitosan. Approved for use in medicine.

Gelatin is a partially hydrolyzed collagen protein. It is used as a gelling component to obtain medicinal products in the form of gels and suppositories.

Beeswax is a waste product of bees with a wide range of medicinal properties and applications. As a thickening additive, it is included in gels and suppositories.

Confectionery fat is refined animal fat. It is used in pharmaceuticals as an ointment and suppository base.

TWIN-80 is a non-ionic surfactant of natural origin, used to stabilize emulsions, including for pharmacological purposes.

Viscoelastic properties are assessed by the magnitude of the elastic modulus (G ') and losses (G ") in the oscillatory mode on a Rheotest RN4.1 rotational viscometer (Germany) with a" cone-cylinder "measurement system at a constant amplitude of 0.01 mNm in the frequency range (ω) 0.5 -10 Hz at 4, 25 and 37 ° С and thermostating time 30 min.

The time of the phase transition "liquid-hydrogel" is assessed visually at 37 ° C by the time of loss of fluidity of the system by inverting the flask. For the time of gelation, the time is taken, after which the system turns into a gel-like state and does not show a flow for 10-15 seconds.

Antibacterial activity is investigated by diffusion in agar on a culture of gram-negative strains (S. typhimurium55) and gram-positive microorganisms (B. cereus 8833,S. aureus 6538). On Petri dishes with sterile agar, 100 μl of a daily broth culture of the microorganism is applied, spread over the surface with a sterile spatula and placed in a thermostat at 37 ° C for 40 minutes. Then, 50 μl of the test preparations are applied to the surface of the cups and thermostated for 24 hours. The result is assessed by the presence or absence of a zone of inhibition of the growth of microorganisms.

The therapeutic and prophylactic efficacy of the funds obtained is tested on sexually mature cows after calving with serous-catarrhal vaginitis. As a traditional comparison therapy, the drug "Intrauterine sticks with ichthyol" (CJSC NPP "Agrofarm") is used. The cows in the experiment are not treated with other drugs. Before treatment, the external genital organs of animals are washed with a 0.1% aqueous solution of potassium permanganate. The drugs are administered intravaginally after the morning milking of animals. Control of treatment is carried out 2 times a day by examination with a vaginal speculum. The dynamics of recovery is assessed according to the following clinical signs: a decrease in exudative discharge, transparency and consistency of the exudate.

The invention is illustrated by photographs, which show:

in fig. 1 - photo of an injectable hydrogel based on chitosan hydrochloride and Pluronic F-127 at 4 and 37 ° C (example No. 2);

in fig. 2 - photo of a suppository suppository based on chitosan hydrochloride, ascorbic acid, Pluronic F-127 and gelatin (example No. 10);

in fig. 3 - photo of a suppository suppository based on chitosan hydrochloride, ascorbic acid, Pluronic F-127, silicon glycerolate in glycerin and gelatin (example No. 11).

The invention is illustrated by tables in which are presented:

in table 1 - component and mass composition, rheological parameters and gelation time of compositions for obtaining injectable hydrogels based on chitosan hydrochloride (examples No. 1-9);

in table 2 - the component and mass composition of the compositions for obtaining suppositories based on chitosan hydrochloride (examples No. 10-13);

in table 3 - antibacterial activity of compositions based on chitosan hydrochloride (example No. 14);

in table 4 - the effectiveness of treatment of cows with postpartum serous-catarrhal vaginitis using compositions based on chitosan hydrochloride (examples No. 15, 16).

in table 5 - the effectiveness of prevention of serous-catarrhal postpartum vaginitis in cows using compositions based on chitosan hydrochloride (examples No. 17, 18).

The method of obtaining funds is as follows. An aqueous solution of chitosan hydrochloride with a concentration of 1.0-3.0 wt. % in distilled water or in an aqueous solution of organic acid with a concentration of 1.0-3.0 wt. % at a molar ratio of chitosan hydrochloride: acid = 1: 1. The process of dissolving the components is carried out according to the standard procedure at room temperature. The resulting solution is introduced into a pharmaceutically acceptable gel base, preferably consisting of (but not limited to) a solution of Pl F-127 and / or gelatin or a mixture of Pl F-127 and gelatin with a glycerol solution of silicon glycerolate or an aqueous-glycerol solution of gelatin or a mixture of confectionery fat and beeswax, mechanically homogenized in a mixer according to the recipe. The resulting composition is poured into syringes to obtain injectable hydrogels or special forms for obtaining a suppository suppository.

The method for preparing an injectable hydrogel is carried out as follows. Pl F-127 is used as a gelling base. For this, a polymer solution is prepared in distilled water at 4 ° C for 24 hours. An aqueous or aqueous-acid solution of chitosan hydrochloride is introduced into the resulting solution at the ratio of components in the finished product chitosan hydrochloride: Pl F-127 = 0.3-2.1: 7.5-18.0, stirred on a mechanical stirrer at a temperature of 20 ± 2 ° C for 5-10 min, poured into injection syringes of a given volume (1-20 ml) and stored at a temperature not exceeding 4-15 ° C. The optimal conditions for the formation of an injectable hydrogel are the conditions under which the “liquid-hydrogel” phase transition occurs at T≥25-37 ° C, while the value of G> Gʺ, and the gelation time ( t ) is no more than 5 minutes.

The method of forming candle suppositories is carried out as follows. As a gel-forming base used: 6-13% aqueous solution of gelatin or a mixture of 6% aqueous solution of gelatin and glycerin in a ratio of 1: 2 or a mixture of 13% aqueous solution of gelatin and 25% aqueous solution of Pl F-127 or their mixture with glycerol solution of glycerolate silicon or a mixture of confectionery fat and beeswax in a ratio of 5: 1. If necessary, an emulsifier is additionally introduced into the system. All used gel bases are preliminarily melted in a water bath at 60-80 ° C for 1-2 hours, an aqueous or aqueous-acid solution of chitosan hydrochloride is introduced at a component ratio of chitosan hydrochloride: gel-forming base = 0.3-3.0: 14.5-77.0, stirred, poured into a mold for making candles and cooled to 4-15 ° C for 24 hours. The finished suppository in the form of a candlestick is placed in a sealed package and stored in a dark place at a temperature not exceeding 15 ° C.

Group of examples No. 1-6. Obtaining injectable hydrogels based on chitosan hydrochloride (table 1).

Example # 1. Prepare 1 wt.% Aqueous solution of chitosan hydrochloride with MW = 30 kDa and 25 wt. % aqueous solution of Pluronic F-127. Solutions of chitosan hydrochloride and the gel-forming base are mixed in a volume ratio of 30:70, stirred on a mechanical stirrer at 20 ° C for 5-10 minutes, filled into injection syringes with a volume of 10 ml and stored at 4 ° C. Composition and content of components, wt%:

chitosan hydrochloride - 0.3;

gelling base (Pluronic F-127) - 17.5;

water is the rest.

The values of the moduli of elasticity (G ') and losses (Gʺ) are: 4 ° C - 0.7 and 0.7 kPa; 25 ° C - 1.0 and 1.0 kPa; 37 ° С - 5.0 and 5.0 kPa, respectively. Gelation of the composition at a temperature of 30 ° C and above does not occur.

Example # 2. All stages are similar to example # 1. The difference is that chitosan hydrochloride with MW = 70 kDa, its 5 wt% aqueous solution and 15 wt% Pluronic F-127 solution are used, which are mixed in a volume ratio of 10:90. Composition and content of components, wt. %:

chitosan hydrochloride - 0.5;

gelling base (Pluronic F-127) - 13.5;

water is the rest.

The values of the moduli of elasticity (G ') and losses (Gʺ) are: 4 ° C - 0.2 and 0.3 kPa; 25 ° С - 0.2 and 0.6 kPa; 37 ° С - 1.5 and 0.2 kPa, respectively. The gelation time ( t ) at 37 ° C is 3 minutes.

Example No. 3. All stages are similar to example # 2. The difference lies in the fact that a 7 wt.% Aqueous solution of chitosan hydrochloride and 20 wt. % Pluronic F-127 solution. Composition and content of components, wt. %:

chitosan hydrochloride - 0.7;

gelling base (Pluronic F-127) - 18.0;

water is the rest.

Values G 'and Gʺ: 4 ° С - 0.1 and 0.1 kPa; 25 ° С - 0.9 and 0.5 kPa; 37 ° С - 1.6 and 0.5 kPa, respectively; t = 3 min.

Example No. 4. All stages are similar to example # 1. The difference is that chitosan hydrochloride with MW = 38 kDa is used, its 3 wt. % aqueous solution and 25 wt. % water solution Pluronic F-127 solution. Composition and content of components, wt. %:

chitosan hydrochloride - 0.9;

gelling base (Pluronic F-127) - 17.5;

water is the rest.

Values G 'and Gʺ: 4 ° С - 1.0 and 1.0 kPa; 25 ° С - 3.5 and 3.2 kPa; 37 ° С - 32.0 and 28.0 kPa, respectively; t = 3 min.

Example No. 5. All stages of the method for obtaining a hydrogel are similar to example No. 4. The difference lies in the fact that D- ascorbic acid is additionally introduced into the composition of the hydrogel at a molar ratio of chitosan hydrochloride: D- ascorbic acid - 1: 1. Composition and content of components, wt. %:

chitosan hydrochloride - 0.9;

D- ascorbic acid - 0.9;

gelling base (Pluronic F-127) - 17.5;

water is the rest.

Values G 'and Gʺ: 4 ° С - 1.0 and 1.0 kPa; 25 ° С - 12.4 and 12.8 kPa; 37 ° С - 15.3 and 13.0 kPa, respectively; t = 3 min.

Example No. 6.

All stages of the method for obtaining a hydrogel are similar to example No. 5. The difference is that boric acid is used. Composition and content of components, wt. %:

chitosan hydrochloride - 0.9;

boric acid - 0.9;

gelling base (Pluronic F-127) - 17.5;

water is the rest.

Values G 'and Gʺ: 4 ° С - 3.0 and 3.0 kPa; 25 ° С - 10.0 and 20.0 kPa; 37 ° С - 78.0 and 25.0 kPa, respectively; t = 3 min.

Example No. 7. All stages of the method for obtaining a hydrogel are similar to example No. 1. The difference is that 3 wt. % aqueous solution of chitosan hydrochloride, solutions of chitosan hydrochloride and Pluronic F-127 are mixed in a volume ratio of 50:50. Composition and content of components, wt. %:

chitosan hydrochloride - 1.5;

gelling base (Pluronic F-127) - 12.5;

water is the rest.

Values G 'and Gʺ: 4 ° С - 5.0 and 5.0 kPa; 25 ° С - 20.0 and 18.0 kPa; 37 ° С - 25.0 and 24.0 kPa, respectively; t = 5 min.

Example No. 8. All stages of the method of obtaining a hydrogel are similar to example No. 3. The difference is that 8 wt. % aqueous solution of chitosan hydrochloride, solutions of chitosan hydrochloride and Pluronic F-127 are mixed in a volume ratio of 20:80. Composition and content of components, wt. %:

chitosan hydrochloride - 1.6;

gelling base (Pluronic F-127) - 16.0;

water is the rest.

Values G 'and Gʺ: 4 ° С - 0.1 and 0.1 kPa; 25 ° С - 0.4 and 0.3 kPa; 37 ° С - 1.3 and 0.5 kPa, respectively; t = 5 min.

Example No. 9. All stages of the method for obtaining a hydrogel are similar to example No. 4. The difference is that solutions of chitosan hydrochloride and Pluronic F-127 are mixed in a volume ratio of 70:30. Composition and content of components, wt. %:

chitosan hydrochloride - 2.1;

gelling base (Pluronic F-127) - 7.5;

water is the rest.

Values G 'and Gʺ: 4 ° С - 3.0 and 3.0 kPa; 25 ° С - 16.0 and 17.0 kPa; 37 ° С - 20.0 and 20.0 kPa, respectively. Gelation at a temperature of 30 ° C and above does not occur.

Thus, based on examples No. 1-9, we can conclude that the optimal ratio for the formation of an injectable hydrogel is the ratio of chitosan hydrochloride: Pl F-127 = 0.5-1.6: 12.5-18.0. In this range of component ratios, the composition at 4 and 25 ° C is a liquid, and at a physiological temperature of 37 ° C it is a form-stable hydrogel.

Group of examples No. 10-13. Obtaining suppository suppositories based on chitosan hydrochloride (table 2).

Example No. 10. The method of obtaining a suppository is similar to example No. 5. The difference is that a 13% solution of gelatin heated to 60 ° C is additionally introduced into the composition in the ratio of the components solution of chitosan hydrochloride in D- ascorbic acid: Pluronic F-127 solution: gelatin solution = 0.3: 0.7: 2.0, mixing of the components is carried out at 60 ° C, the resulting composition is poured into a mold for making candles and cooled to 4-15 ° C for 24 hours. The finished suppository in the form of a candlestick is placed in a sealed package and stored in a dark place at a temperature not exceeding 15 ° C. Composition and content of components, wt%:

chitosan hydrochloride - 0.3;

D- ascorbic acid - 0.3;

gelling base (Pluronic F-127 + gelatin) - 14.5;

water is the rest.

Example No. 11.

All stages of obtaining a suppository suppository are similar to example No. 10. The difference is that a glycerol solution of silicon glycerolate is additionally introduced into the composition in the ratio of components solution of chitosan hydrochloride in D- ascorbic acid: Pluronic F-127 solution: gelatin solution: silicon glycerolate solution = 0.3: 0.7: 1: 2. Composition and content of components , wt%:

chitosan hydrochloride - 0.3;

D- ascorbic acid - 0.3;

gelling base (Pluronic F-127 + gelatin + glycerin solution of silicon glycerolate) - 29.0;

water is the rest.

Example No. 12. The stages of obtaining a suppository suppository are similar to example No. 10. The difference is that a 6% water-glycerol solution of gelatin heated to 70 ° C is used as a gel- forming base, chitosan hydrochloride and D- ascorbic acid are introduced into the gel-forming base in the form of a mixture of dry powders, the components are mixed at a temperature of 70 ° C. Composition and content of components, wt%:

chitosan hydrochloride - 3;

D- ascorbic acid - 3;

gelling base (glycerin + gelatin) - 40.0;

water is the rest.

Example No. 13.

All stages of obtaining a suppository suppository are similar to example No. 10. The difference is that a mixture of confectionery fat and beeswax (5: 1) heated to 80 ° C is used as a gel-forming base, the components are mixed in the ratio of a solution of chitosan hydrochloride in D- ascorbic acid: a mixture of confectionery fat and beeswax = 1.0: 4.0, the emulsifier TWIN-80 is additionally introduced, the mixing of the components is carried out at a temperature of 80 ° C. Composition and content of components, wt. %:

chitosan hydrochloride - 0.6;

D- ascorbic acid - 0.6;

gel-forming base (confectionery fat + beeswax) - 77.0;

emulsifier TVIN-80 - 3;

water is the rest.

It has been experimentally established that when used in a suppository suppository the ratio of the components of chitosan hydrochloride: gelling base = 0.3-3.0: 14.5-77.0, stated in examples No. 10-13, the result of the invention is achieved.

Example No. 14. Evaluation of the antibacterial activity of compositions based on chitosan hydrochloride (table 3).

In sterile Petri dishes with sterile agar, add 100 μl of a daily broth culture of microorganisms B. cereus 8833 or S. aureus 6538 or S. typhimurium and incubated at 37 ° C for 40 minutes. Then, on the surface of the cups, 50 μl of injectable hydrogels obtained according to examples No. 4-6 are applied and thermostated for 24 hours. Antibacterial activity is assessed by the presence or absence of a zone of inhibition of the growth of microorganisms.

As can be seen from the data in Table 3, all tested agents exhibit antibacterial activity against all tested cultures. The greatest antibacterial activity is observed in the case of gram-positive microorganisms ( B. cereus 8833, S. aureus 6538), as well as gram-negative microorganisms ( S. typhimurium ) for the composition obtained according to example No. 6.

Examples No. 15-18. Treatment and prevention of serous-catarrhal vaginitis using compositions based on chitosan hydrochloride (table 4, 5).

Example No. 15.

Determination of the therapeutic effect of the compositions based on chitosan hydrochloride was carried out on black-finger cows at the age of 3-4 years suffering from serous-catarrhal vaginitis. The research was carried out on the collective farm "Zarya" of the Tamalinsky district of the Penza region. The experimental group of animals (5 cows) used an injectable hydrogel according to example No. 5 at a dosage of 1 syringe per 10 ml once a day for 3 days. The animals of the control group (5 cows) were treated with the traditional preparation "Intrauterine sticks with ichthyol" (manufactured by CJSC NPP "Agrofarm") according to the instructions.

Positive dynamics in animals treated with an injectable hydrogel was noted on the 2nd day of treatment and reached 40% of recovered animals. In the control on the 2nd day of treatment, only 20% of recovered animals were observed. Full recovery of the animals in the experimental group was observed on the 3rd day of treatment, the control group - on the 5th day.

Example No. 16. All stages of treatment of cows with serous-catarrhal vaginitis (experimental group and control) are similar to example No. 15. The difference is that the treatment of animals in the experimental group was carried out using suppositories according to example No. 12.

Positive dynamics in animals treated with a suppository suppository was noted already on the first day of treatment and reached 20% of those who recovered. On the second day of treatment, the number of recovered animals was 60%. Full recovery (100% of recovered animals) was observed on the 3rd day of treatment.

Example No. 17. All stages of prophylactic therapy of cows, aimed at preventing the appearance of serous-catarrhal vaginitis, are similar to example No. 15. The difference is that the therapy is carried out on cows 2-3 days after calving (after closing the cervix), which still do not show signs of serous-catarrhal vaginitis, with a single use of the drug.

In the experimental group, no animals with acute serous-catarrhal vaginitis were observed for 5 days of observation. In the control on the 2nd day, 60% of sick animals were observed. During the next 3 days of observation, the percentage of diseased animals in the control group did not change. In the future, the control group of animals used drug treatment with antimicrobial agents ("Nitox 200", "Ditrim") until complete recovery.

Example No. 18. All stages of preventive therapy for cows are similar to example No. 17. The difference is that the therapy was carried out using suppositories obtained according to example No. 12. In the experimental group, on the 3rd day of observation, 20% of sick animals were noted. During the next 3 days of observation, the percentage of diseased animals in the control group did not change. In the future, the animals of the experimental group were treated similarly to example No. 16 until complete recovery.

From examples No. 15, 16 and table 4 it can be seen that in the groups of animals, the treatment of which was carried out using a composition based on chitosan in the form of an injectable hydrogel or suppository suppository, the efficiency of recovery is significantly higher compared to standard therapeutic treatment.

From examples No. 17, 18 and table 5, it can be seen that in groups of animals whose preventive therapy was carried out using a composition based on chitosan hydrochloride in the form of an injectable hydrogel or suppository suppository, a higher prophylactic effect is observed in comparison with standard therapy.

In addition, the treatment and prevention of serous-catarrhal vaginitis in cows using the developed means based on chitosan hydrochloride does not impose restrictions on the use of milk and animal meat for food purposes both during and in the first few days after treatment.

Table 1

Component and mass composition, rheological parameters and gelation time of compositions for obtaining injectable hydrogels based on chitosan

Reagents Content of starting components, wt% Examples, no. one 2 3 4 five 6 7 eight nine Hydrochloride
chitosan MM, kDa 0.3 - - - - - 1.5 - - thirty 38 - - - 0.9 0.9 0.9 - - 2.1 70 - 0.5 0.7 - - - - 1.6 - D-Ascorbic acid - - - - 0.9 - - - - Boric acid - - - - - 0.9 - - - Pl F-127 17.5 13.5 18.0 17.5 17.5 17.5 12.5 16.0 7.5 Water rest Rheological parameters of hydrogels, kPa T = 4 ° C G ' 0.7 0.2 0.1 1.0 1.0 3.0 5.0 0.1 3.0 Gʺ 0.7 0.3 0.1 1.0 1.0 3.0 5.0 0.1 3.0 T = 25 ° C G ' 1.0 0.2 0.9 3.5 12.4 10.0 20.0 0.4 16.0 Gʺ 1.0 0.6 0.5 3.0 12.8 20.0 18.0 0.3 17.0 T = 37 ° C G ' 5.0 1.5 1.6 32.0 15.3 78.0 25.0 1.3 20.0 Gʺ 5.0 0.2 0.5 28.0 13.0 25.0 24.0 0.5 20.0 t, min - 3 3 3 3 3 five five -

table 2

Component and mass composition of compositions for producing suppositories based on chitosan

Reagents Content of starting components, wt% Examples, no. 10 eleven 12 13 Chitosan hydrochloride
MM = 38 kDa 0.3 0.3 3.0 0.6 D-Ascorbic
acid 0.3 0.3 3.0 0.6 Pl F-127 5.8 5.8 - - Gelatin 8.7 6.5 24.0 - Silicon glycerolate - 4.2 - - Glycerol - 12.5 16.0 - Confectionery fat - - - 65 Beeswax - - - 12 TWIN-80 - - - 3 Water rest

Table 3

Antibacterial activity of compositions based on chitosan

Bacterial strain
test cultures Antibacterial activity The sample obtained according to example No. 4 five 6 B. cereus 8833 ++ ++ ++ S. aureus 6538 ++ ++ ++ S. typhimurium 55 + + ++

+ - moderate antibacterial activity;

++ - high antibacterial activity.

Table 4

The effectiveness of treatment of cows with postpartum serous-catarrhal vaginitis using compositions based on chitosan

Treatment period, days The number of recovered animals,% Examples, no. fifteen sixteen The control one - twenty - 2 40 60 twenty 3 100 100 60 five - - 100

Table 5

Efficiency of prevention of serous-catarrhal postpartum vaginitis in cows using compositions based on chitosan

Observation period, days The number of sick animals,% Examples, no. 17 18 The control one - - - 2 - - 60 3 - twenty 60 five - twenty 60

Claims (2)

An agent for the prevention and treatment of vaginitis in cows, containing an active and auxiliary part, characterized in that it contains chitosan hydrochloride with a molecular weight of 30-70 kDa and ascorbic or boric acid as an active part, and a gel-forming base chosen from a triblock copolymer of ethylene oxide and propylene oxide, or a mixture of gelatin with glycerin, or a triblock copolymer of ethylene oxide and propylene oxide with gelatin, or a triblock copolymer of ethylene oxide and propylene oxide with gelatin and tetraglycerolate silicon, or a confectionery fat and non-beeswax surfactant the following ratio of components, wt%: chitosan hydrochloride 0.3 - 3.0 ascorbic or boric acid 0.3 - 3.0 gelling base 12.5 - 77.0 water rest

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