RU2391110C2 - Medication and method of treating necrotic suppurative diseases of animal extremities - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to field of veterinary, in particular to means and methods of treating necrotic suppurative diseases of animal extremities, such as necrobacteriosis, footrot, suppurative pyodermatites, laminites, Rustergolts ulcers, etc. Claimed medication represents nanostructured composition in form of suspension, containing clathrate of didecyldimethylammonium halagenide with carbamide, copper vitriol, tar, pharmaceutically acceptable monoatomic alcohol and water. Also described is method of treating necrotic suppurative diseases of animal extremities which lies in the following: extremities of animals are treated with efficient amount of medication 1 time a day during 7-10 days in form of external applications and/or foot baths.

EFFECT: medication possesses high penetrating ability into affected tissues of organism, does not damage healthy cells, demonstrates high activity with respect to wide spectrum of microorganisms both in vitro and in vivo.

5 cl, 1 tbl, 8 ex

Description

The invention relates to the field of veterinary medicine, in particular to means and methods for treating purulent-necrotic diseases of animal limbs, such as necrobacteriosis, hoof rot, purulent pyodermatitis, laminitis, Roostergolts ulcers, etc.

Infectious diseases of the extremities are affected by all age and sex groups of animals. According to some data, the incidence in the Russian Federation in various farms varies widely from 12 to 88%. In this case, the most common disease is necrobacillosis.

The leading role in the pathological process is played by Fusobacterium necrophorum (formerly Sphaerophorus necrophorus) and, less commonly, Dichelobacter nodosus (formerly Bacteroides nodosus) - anaerobic toxin-forming microorganisms that cause subacute or acute course of necrobacteriosis of the hoof from one of the skin pathogens or and deeper tissues. With the progression of necrobacteriosis and chronicity of the process, deeper structures of the limbs may be affected, and in advanced cases, joints, ligaments, as well as mucous membranes and internal organs may be involved in the process.

Over the past three years, the number of cases of cattle necrobiosis, according to official statistics, has doubled in Russia.

In this regard, the issue of the development of effective anti-fusobacterial agents raises the issue.

Currently, inactivated associated vaccines against necrobacteriosis of cattle limbs are registered in the Russian Federation (the patented names are “Nekovak”, “Nekovak-Stimulus”, patent of the Russian Federation 2098127 dated 10.12.1997) and an inactivated associated vaccine against infectious diseases of the limbs of sheep (patented name "Ovikon", patent of the Russian Federation 2098128 dated 12/10/1997). However, experience with vaccines did not show their high prophylactic activity. This is due to the characteristics of the biology of the pathogen, pathogenesis and immunogenesis of the disease.

Various veterinary drugs have also been developed to combat infectious diseases of the lower extremities of animals.

There is a method of treating hoof rot of sheep and goats, including the passage of animals through a bath with a solution of copper sulfate, and before the run of animals, they are driven through a bath with an aqueous solution of anionic surfactant, and after the bath with copper sulfate, through a bath with an aqueous solution of a mixture of Quaternary ammonium salts. A 0.1-0.25% aqueous solution of C11-C17 sodium alkyl sulfonate is used as an aqueous solution of an anionic surfactant, and a mixture of the following composition, wt.%: C10-C18 alkyl trimethylammonium, is used as a mixture of quaternary ammonium salts chloride 2.5-5.0; cetylperidinium chloride 0.5-1.0; water is the rest.

The disadvantage of this method is the low therapeutic effect, the complexity, as well as the short duration of the effect of the medicinal product on the affected area.

A known tool for the treatment of purulent-necrotic diseases of animal limbs for individual foot baths on affected limbs of animals, which is an ointment of the following composition, wt.%:

Veterinary Copper Sulphate 7.5-15 Birch tar 9-13 Ichthyol 3-5 Petroleum jelly rest

which is prepared at 36-40 ° C with mechanical stirring, and the duration of the ointment on the affected area is 3-10 days (RF Patent 2025110, publ. 1994.12.30).

However, with this method of treatment, the effectiveness of treatment is low due to the low degree of penetration of the ointment in the affected area.

Also known is an aqueous composition based on a copper salt, also containing a quaternary ammonium compound and peroxide in the following ratio of components, wt.%: Copper salt 2-20; quaternary ammonium compound 0.5-2; peroxide 0.5-5; water the rest. A preferred quaternary ammonium compound in the composition of this invention is a mixture of N-alkyl (C12-18) -N chloride, N-dimethylbenzylammonium chloride and N-alkyl (C12-18) -N chloride, N-dimethylethylbenzylammonium sold under the trade name BTC 2125 M offered by Stepan (Northfield, Illinois). Another preferred quaternary ammonium compound in the composition of this invention is a mixture of N-alkyl (C12-18) -N chloride, N-dimethyl-N-benzylammonium, N-dodecyl-N, N-dimethyl-N-ethylbenzylammonium chloride and N- tetradecyl-N, N-dimethyl-N-benzylammonium sold under the trade name Barquat 4280-2 by Lonza (Fair Lawn, New Jersey) (RF patent 2235546, publ. 2004.09.10).

This technical solution can be considered as the closest analogue (prototype). The known composition does not have a sufficiently effective effect.

The lack of effectiveness of existing drugs for the treatment of purulent-necrotic diseases of animal limbs is associated with a number of features of these diseases.

In diseases of the extremities, the affected organ is constantly in contact with the soil and other environmental objects, which leads to its abundant contamination with various bacterial flora; in addition, one of the characteristic features of anaerobic infections is the simultaneous associative effect on the body of two, three or more types of anaerobic and aerobic microorganisms (Baltrashevich A.K., 1979; Sidorchuk A.A. et al., 2006; Katie RV et al., 1976). The large role of concomitant microflora, which significantly complicates the course of the disease.

In addition, the main causative agent of necrobiosis Fusobacterium necrophorum is a normal component of ruminant rumen microflora. Based on this, it can be assumed that the pathogen has the ability to change in accordance with environmental conditions, forming pathogenic and harmless strains. Therefore, it is necessary to fight not only with the pathogen, but also with the symptoms that arose as a result of improper keeping of animals.

It can also be concluded that pathogens must be destroyed during the initial introduction into the tissue of the inter-experimental gap, when the non-pathogenic strain passes into the pathogenic one.

The objective of the invention is to increase the effectiveness of the treatment of infectious diseases of the lower extremities of animals, especially necrobacteriosis, by developing a tool with a wide spectrum of antimicrobial action with residual antimicrobial effect, which has high penetration into the affected tissues of animals that does not damage healthy cells, which can be used repeatedly.

The problem is solved with the help of a new generation of therapeutic and prophylactic agents, which is a suspension nanostructured composition containing didecyldimethylammonium halide clathrate with urea, copper sulfate, tar, pharmaceutically acceptable monohydric alcohol and water with the following components in wt.%:

didecyldimethylammonium clathrate carbamide halide 2-10 copper sulfate 1-10 tar 2-10 monohydroxy alcohol 2-10 water rest

moreover, the average particle size is from 10 to 100 nm.

The didecyldimethylammonium halide clathrate with urea included in the composition is a known disinfectant that exhibits a tonic, cleansing, anti-inflammatory, antiseptic, antibacterial, antiviral, antifungal effect. Didecyldimethylammonium halide clathrates with carbamide (CDHC) can be presented as chlorides, bromides and iodides.

Tar birch (Pix Betulae, Oleum Rusci) is a preparation obtained by dry distillation of the outer part of the birch bark (selected birch bark). Contains benzene, xylene, cresol, cresot, toluene, guaiacol, phenol, resins and other substances, has strong antiseptic, locally irritating, keratoplastic (3-5%), keratolytic (30-50%), anti-inflammatory, insecticidal, antiparasitic and disinfecting properties , improves blood supply to tissues, moderately irritates sensitive nerve endings, stimulates the regeneration of the epidermis of damaged tissues, enhances the keratinization process, wounds wounds and accelerates their healing. After a short exposure, it kills many bacteria, in particular tubercle bacillus and anthrax spores. In addition, as part of the proposed tool, it is used to consolidate the composition in the cultivated areas.

Copper sulfate is used in veterinary medicine as an antiseptic.

As the monohydric alcohol, it is preferable to use alcohols such as isopropyl or ethyl alcohol.

The tool can be used to treat various types of animals, including cattle, sheep, goats, with lesions of the extremities by external applications and / or foot baths. For therapeutic purposes, the drug is used 1 time per day for 7-10 days. They process wound surfaces, as well as hooves and soles of limbs. For prophylactic purposes, with planned trimming of hooves, limbs are treated once. The therapeutically effective amount depends on the type of animal, the degree of damage, etc.

The form of the substance in the form of nanosomes provides selective accumulation in the foci of infectious lesions. Due to this, the drug practically does not enter the systemic circulation, ensuring the absence of toxicity, while the duration of the drug increases, creating a depot in the focus of the pathological process.

The method for producing the agent consists in mixing an alcoholic solution of didecyldimethylammonium halide clathrate with carbamide and copper sulfate, adding the resulting mixture to birch tar, followed by intensive mixing of the mixture.

To obtain a nanostructured composition with the indicated particle size, methods known in the field of nanotechnology can be used, for example, the use of ultrasonic treatment, high pressure homogenization, etc.

Particle sizes were determined by autocorrelation spectroscopy on a Coulter N4MD submicron laser spectrometer manufactured by Coulter Electronics (France - USA).

The possibility of carrying out the invention is demonstrated by the following examples.

Example 1

didecyldimethylammonium bromide clathrate with urea 10 copper sulfate 2.5 tar 5 isopropyl alcohol 10 water 72.5

Example 2

didecyldimethylammonium chloride clathrate with urea four copper sulfate one tar 5 isopropyl alcohol four water 86

Example 3

didecyldimethylammonium bromide clathrate with urea 2 copper sulfate 10 tar 10 ethanol 2 water 76

Example 4

didecyldimethylammonium bromide clathrate with urea 10 copper sulfate 5 tar 5 isopropyl alcohol 10 water 70

Example 5

didecyldimethylammonium bromide clathrate with urea 5 copper sulfate 0.5 tar 2 isopropyl alcohol 5 water 87.5

Example 6

Determination of activity against Fusobacterium necrophorum.

To determine the bactericidal activity of the preparation, blood agar was seeded with a continuous lawn Fusobacterium necrophorum isolated from necrotic tissue and wounds of cows, paper disks impregnated with the compositions of Examples 1-5, and also a suspension containing only didecyldimethylammonium bromide clathrate with isopropyl carbamide and water, cultivation was carried out in an anaerostat for 24 hours at a temperature of 37 ° C. After incubation, the surface of the medium and the grown culture were viewed in oblique light and ryali zone of no growth of the microorganism under study. It was found that the zone of lack of growth when using the compositions according to examples 1-5 was 3.4-3.8 cm, which indicates a high activity of the claimed funds in relation to the pathogen. The growth zone when using the control composition was 2.6 cm

Example 7

Tests of therapeutic activity

In production tests, 270 cows with necrobacteriosis of the cow were used.

Animals were divided into experimental and control groups. The animals of the experimental groups (5 groups) were treated with the claimed agent according to examples 1-5 daily 1 time per day for 10 days. Before applying the preparation, the hooves were cleaned of dirt and, if necessary, the hoofed horn was cut with a hoofed knife. The control animals were treated with the well-known commercial antibacterial agent Terramycin.

On the second or third day of treatment of the cows of the experimental groups, the swelling of the skin and subcutaneous tissue decreased markedly, the ulcerated skin areas dried out, and the outflow from the wounds decreased. On days 4-5, the signs of inflammation disappeared, the wounds were healed, and a dense scab formed. On the 7th day, the limp of the supporting limbs decreased, and the signs of necrobiosis completely disappeared. In cows of the control group, the healing process was 2 times slower.

Tests were also conducted on 48 sheep affected by hoof rot.

The tool was used in the form of foot baths 1 time per day for 7 days. Sheep was placed for 15 minutes in a metal container filled with the product. The level of the upper meniscus was 5 cm.

Daily, the sheep were examined and the physiological state of the body was taken into account.

On 4-5 days in the sheep, swelling decreased. Wounds dried up and the wounds healed. On the 7th day, the limp of the supporting limbs disappeared. The signs of hoof rot have completely disappeared. All sheep, regardless of the severity of the disease, recovered.

Studies have been conducted on milk and meat of animals treated. It was found that the components of the product were not excreted in milk and did not affect the quality of meat.

Example 8

The study of the stability of the product during storage.

The stability of the properties of the product during storage was determined by the method of “accelerated aging”, based on the increase in the rate of chemical processes at elevated temperatures exceeding the maximum allowable temperature during storage of the product under normal conditions.

For this purpose, 3 series of experimental samples in a volume of 300 cm ³ were placed in a thermostat with a temperature of 50 ° C. The quality of the series was evaluated by changing the mass fraction of the ammonium clathrate compound with urea, the density and refractive index in accordance with STO 719544720-0003-2008.

The experiment was carried out for 12 months. Every 2 months, samples were taken and the mass fraction of the active substance was determined by two-phase titration using an anionic standard solution (sodium dodecyl sulfate) in an alkaline medium in the presence of a blue bromophenol indicator, density using a hydrometer, and refractive index using a refractometric method.

The results of the study are presented in table 1.

Table 1 Duration of storage, month Year corresponding to the shelf life Mass fraction of active substance,% Density g / cm ³ Refractive index ONE HUNDRED Property Stability 2 one 10 1,055 1,353 Acc. Stab. four 2 10 1,055 1,353 Acc. Stab. 6 3 10 1,055 1,353 Acc. Stab. 8 four 10 1,055 1,353 Acc. Stab. 10 5 10 1,055 1,353 Acc. Stab. 12 6 10 1,055 1,353 Acc. Stab.

Thus, the shelf life of the product is at least 6 years.

The proposed tool is highly effective and can be recommended for widespread implementation in veterinary practice.

Claims (5)

1. An agent for external use for the treatment or prevention of purulent-necrotic diseases of animal limbs, containing a copper salt, a quaternary ammonium compound and water, characterized in that it is a nanostructured composition in the form of a suspension with an average particle size of from 10 to 100 nm, including as a quaternary ammonium compound, a cladrate of didecyldimethylammonium halagenide with urea, copper sulfate as a copper salt, and also tar and a pharmaceutically acceptable noatomny alcohol with the following contents, wt.%:
didecyldimethylammonium halagen clathrate with urea 2-10 copper sulfate 1-10 tar 2-10 monohydroxy alcohol 2-10 water rest 2. The tool according to claim 1, characterized in that it contains didecyldimethylammonium halide with urea as a clathrate; didecyldimethylammonium bromide with urea clathrate. 3. The tool according to claim 1 or 2, characterized in that it contains isopropyl alcohol as the monohydroxy alcohol. 4. A method of treating purulent-necrotic diseases of animal limbs, characterized in that the affected areas of the animals are treated with an effective amount of the agent according to any one of claims 1 to 3 1 time per day for 7-10 days by external applications and / or baths. 5. The method according to claim 4, characterized in that said disease is necrobacteriosis.