RU2400218C1 - Method of increasing efficiency of antibiotics - Google Patents

Abstract

FIELD: medicine. ^ SUBSTANCE: invention refers to preparation of antibiotics. The method of increasing the efficiency of antibiotics includes detoxification and polymerisation of antibiotics with 0.150.05% formalin solution at 40.02.0C for 5-7 days followed by preparation of liquid or lyophilised drugs. ^ EFFECT: method according to the invention allows increasing the efficiency of antibiotics by 5010% with simultaneous decrease in their toxicity and allergenic capacity that allows using antibiotic preparations in a medical range with higher therapeutic effect with reduced contra-indications. ^ 1 tbl, 4 ex

Description

The invention relates to microbiology and biotechnology, in particular to the production of antibiotics, including beta-lactam.

It is known that the number of bacteria producing beta-lactamose enzymes that break down antibiotics is constantly increasing.

Long-term use of antimicrobials and exposure to physical and chemical mutagens led to the emergence of antibiotic-resistant microorganisms. Enzymatic cleavage of the antibiotic provides more than 80% bacterial resistance.

To increase the effectiveness of antibiotics, fluorine, piperazine radical are introduced into their chemical structure or combined with other antibiotics: subbactam with ampicillin, succinic, citric, fumaric and other organic acids and their salts - succinates, probiotics (Andreeva N.L., Voitenko V.D. Improving the effectiveness of chemotherapy using organic acids // International Bulletin of Veterinary Medicine. - 2004. - No. 1. - P.55-58.)

The greatest antibacterial effect was obtained by the combination of antibiotics with clavulanic acid, obtained in 1976 from the metabolic product of the fungus Streptomyces clavuligeris. Despite the fact that clavulanic acid itself is a weak antibiotic, it turned out to be an active beta-lactamase inhibitor as a result of penetration into the structure of beta-lactamase and blocking its activity (Korotyaev A.I., Babichev S.A. Medical microbiology, immunology and virology . / Textbook for high schools. - St. Petersburg .: Special literature, 1998. - S.135-142).

The prototype is a combination of amoxicillin and clavulanic acid (Pokrovsky V.I., Pozdeev O.K. Medical Microbiology. - M .: Publishing House GEOTAR, 1999. - P.148-151).

The disadvantages of this method are the high cost and deficiency of the drug, toxicity to the liver, kidneys, the ability to allergize the body, the emergence of drug-resistant microorganisms, sensitivity to an acidic environment, the creation of dysbiosis in the gastrointestinal tract.

To eliminate these drawbacks, to increase the effectiveness of antibiotics, reduce their toxicity and allergize the body, increase resistance to the degrading effect of bacterial enzymes, create a stable structure of antibiotics, polymerization and detoxification of a number of antibiotics using 0.15 ± 0.05% formalin solution at 40 ± 2.0 ° C for 5-7 days.

The legitimacy of using formalin for the polymerization and detoxification of antibiotics - a product of metabolism and destruction of fungi - is justified by the production and use of toxoids, tolerogens (allergoids), toxoid vaccines, inactivated viral vaccines, etc. However, the use of formalin for detoxification, polymerization of the structure and increasing the effectiveness of antibiotics has not been carried out.

The aim of the invention is to increase the effectiveness of antibiotics against resistant microorganisms, gain resistance to the enzymatic action of bacterial enzymes, reduce toxicity and allergenicity to the body.

The goal is achieved by polymerization and detoxification of antibiotics with 0.15 ± 0.05% formalin solution at 40.0 ± 2.0 ° C for 5-7 days. In this case, the initial concentration of formalin is reduced to 0.05% in a liquid solution, and in a lyophilized form, to 0.01% concentration. The resulting antibiotics gain increased effectiveness against drug-resistant microorganisms, resistance to bacterial enzymes, while reducing toxicity and allergenicity to the body.

In the patent and scientific and technical literature, no technical solutions similar to the declared one were revealed.

The use of 0.15 ± 0.05% formalin solution allows to achieve stable detoxification, polymerization of the entire complex of antibiotics and the production of harmless drugs with increased bacteriostatic and bactericidal action against drug-resistant microorganisms, i.e. get a technical result.

The established ability of a 0.15 ± 0.05% formalin solution at 40.0 ± 2.0 ° C for 5-7 days to provide polymerization, detoxification, structural stability, reduced toxicity and allergenicity, and a 50 ± 10% increase in the effectiveness of antibiotics with regard to drug-resistant microorganisms, it allowed to propose a rational technology for the manufacture of effective antibiotics that are harmless to humans and animals.

Under the action of a formalin solution with a concentration of 0.15 ± 0.05%, the process of polymerization of the structure of antibiotics occurs, as a result of which their resistance to the enzymatic action of bacterial enzymes increases. Formalin also provides complete detoxification of antibiotics.

The indicated concentration of 0.15 ± 0.05% formalin and the temperature regime of treatment are optimal to ensure stable polymerization and detoxification of antibiotics. As a result of this treatment, the modified antibiotics showed increased by 50 ± 10% bactericidal activity in relation to the studied microorganisms, as well as harmlessness due to a decrease in the toxicity of antibiotics. A similar effect of formalin on the detoxification processes of toxins of microorganisms was the basis for the manufacture of modified antibiotics.

The invention is illustrated by the following examples.

Example 1. The method of detoxification and polymerization of antibiotics with 0.15 ± 0.05% formalin solution.

We used 60 vials of lyophilized penicillin, erythromycin, kanamycin, tetracycline, gentamicin, polymyxin, neomycin, amoxiclav, into which 5.0 ml of a 0.2% formalin solution was added by syringe for detoxification and polymerization at 42 ° C for 7 days At the end of detoxification and polymerization, half of the bottles of each antibiotic were left to be studied in liquid form, and the second part was lyophilized.

Example 2. Determination of the residual formalin content in antibiotic solutions before and after lyophilization.

Using titration with a solution of iodine, sodium hydroxide, sulfuric acid and hyposulfite, it was found that formalin levels decreased from 0.2% to 0.05% in liquid antibiotic solutions, and after lyophilization, the formalin content was 0.01% instead of 0.2% ("Pharmacopoeia article" - Physico-chemical and immuno-biological methods of drug control. Official publication of the Ministry of Health of the Russian Federation. - M. 2000 - P.51-56).

Example 3. Determination of the safety of modified antibiotics.

The test was conducted on 10 white mice, 6 guinea pigs, 20 calves and 22 piglets.

After subcutaneous administration to 5 white mice of modified penicillin and 5 mice - polymyxin in 0.2-0.3 ml preparations three times, the animals did not show pronounced toxic effects and death when observed within 12 days.

When subcutaneously administered five times with an interval of 1 day to 5 calves of penicillin, 5 calves of gentamicin, 5 calves of tetracycline and 5 calves of polymyxin, all animals remained alive, no purulent necrotic lesions were noted at the injection site for 20 days (observation period).

Subcutaneous administration to 6 guinea pigs of modified penicillin in a volume of 2-3 ml three times with an interval of 24 hours also did not cause the formation of purulent abscesses at the injection site and the death of animals upon observation for 15 days.

Example 4. The determination of the effectiveness of the obtained antibiotics was determined by the method of serial dilution of drugs. The obtained antibiotics indicated by the letter M (modified) were compared with their corresponding commercial preparations.

The bactericidal properties of antibiotics were determined against Escherichia, staphylococcus, Salmonella, Vas. Subtilis and Proteus. The results are presented in the table.

Comparative evaluation of the effectiveness of antibiotics (U / ml, μg / ml) in a liquid medium containing 10,000 microbial cells in 1 ml No. p / p Preparations Name of microorganisms E.coli S.aureus S.dublin You. Subtilis Proteus Vulgaris one. Penicillin 10-12 12-15 12-14 15-16 15-20 2. M-penicillin 5-7 9-12 8-9 10-12 10-12 3. Gentamicin 10-12 12-13 10-12 13-15 15-16 four. M-gentamicin 4-6 9-10 5-6 9-11 10-12 5. Tetracycline 20-25 20-22 22-25 25-30 25-27 6. M-tetracycline 15-17 15-17 17-18 15-16 15-17 7. Amoxiclav 8-9 7-8 10-12 12-15 12-15 8. M-amoxiclav 3-4 4-6 7-8 9-10 9-10 9. Neomycin 20-22 22-25 20-25 25-30 30-35 10. M-neomycin 10-15 15-17 15-17 15-17 15-17 eleven. Polymyxin 50-60 50-60 50-60 60-65 60-65 12. M-polymyxin 30-40 35-40 35-40 35-40 35-40 13. Erythromycin 8-10 10-12 10-12 12-15 13-15 fourteen. M-erythromycin 5-6 7-8 7-8 9-10 9-10 fifteen. Kanamycin 10-15 10-12 13-15 13-15 15-17 16. M-kanamycin 7-8 6-7 7-8 8-9 9-10 M - modified antibiotic

From the data presented in the table, it follows that antibiotic solutions subjected to detoxification, polymerization with 0.2% formalin solution at 42 ° C for 7 days showed an increased by 50 ± 10% bactericidal effect on E. coli, S.aureus, S .dublin, you. Subtilis and Pr. Vulgaris compared to control antibiotics.

Claims (1)

A method of increasing the effectiveness of antibiotics, characterized in that the polymerization and detoxification of antibiotics is carried out by treatment with 0.15 ± 0.05% formalin solution at 40.0 ± 2.0 ° C for 5-7 days and they are used in liquid or lyophilized condition.