The object of the study is the carbonic acid hop extract, new ointment compositions based on EXB. The aim of the study is to provide a microbiological justification for the development of an ointment composition based on carbonic acid hop extract to improve the effectiveness of treatment of infected wounds. Research methods: microbiological; cytological; technological; mathematical and statistical. Theoretical and practical results: Antimicrobial activity of EXB has been characterized by double serial dilutions according to MIC, MBC and MFC indicators on microorganism test strains; by sequential multiple passages of microorganisms on nutrient media in subinhibitory concentrations the rate of resistance formation to EXB in dominant pathogens of wound infection has been studied; The antimicrobial action of experimental samples of ointment compositions on the basis of ECB and auxiliary substances of museum and clinical isolates was determined by the "wells" method. It was found that antimicrobial activity of EXB for test strains of microorganisms of different taxonomic groups ranged from 1.69 mg/ml (S. aureus ATSS 25923) to 3.40 mg/ml (P. aeruginosa ATSS 27853) and 4.15 mg/ml for C. albicans ATSS 885-653, IBK varied from 3.38 to 7.14 mg/ml respectively, and IFC was 8.53 mg/ml. The cytotoxicity of EXB on cultures of transplanted human laryngeal adenocarcinoma epithelial cell lines (Hep-2) and human diploid fibroblasts (M-19) was determined by the minimum toxic concentration (MTK in mg/ml), the cytotoxic concentration that causes 50% cell death (CK50 in mg/ml) and the selectivity index (SE). In the HER-2 and M-19 cell lines, initial cytotoxic effects were found to be induced by concentrations of EXB ≥5.0 and ≥10.0 mg/mL with corresponding CK50 values of 28.7 and 77.3 mg/mL. In this case, taking into account the MIC of EXB for microorganisms of different taxonomic groups, the value of its IC ranged from 6.9 to 45.7, with the highest level of 17.0 and 45.7 for S. aureus ATSS 25923 and the lowest of 6.9 and 18 6 for C. albicans ATSS 885-653. Using technological methods, experimental samples of ointment compositions based on EXB and excipients were produced. A new antimicrobial composition in the form of ointment was developed (containing per 100 g. EXB - 5,0; propylene glycol - 10,0; emulsifier 1 - 5,0; PEO 1500 - 28,0; PEO 400 - 52,0) with high antimicrobial activity against dominant types of wound infection agents (patent for utility model № 110799 UA "Therapeutic and cosmetic phytomedicines for local application"). Novelty: for the first time the antimicrobial activity of EXB was characterized on the reference microorganism test strains by MIC, MIC and IBC, which values were 1.69 - 4.15 mg/ml, 3.38 - 7.14 mg/ml and 8.53 mg/ml, respectively. The slow formation of resistance to the action of increasing concentrations of EXB in clinical strains of S. aureus and P. aeruginosa at 30 passages of the final MIC level (median) for S. aureus - 11.83 mg / ml and P. aeruginosa - 45.3 mg / ml was proved for the first time. For the first time, we obtained the results of studies that determined the cytotoxicity of EXB on linear cultures of human epithelial cells (Hep-2) and fibroblasts (M-19), which showed a low cytotoxic effect of EXB on eukaryotic cells (GC 50 28.7 and 77.3 mg / ml, respectively) and its high selectivity (IC 17.0 - 45.7 for S. aureus ATSS 25923 and P. aeruginosa ATSS 27853 and 6.9 - 18.6 for C. albicans ATSS 885-653). For the first time eight experimental samples (Nos. 1-8) of ointment compositions with EXB and excipients in different concentrations were developed, which fully meet the current requirements of GFU 2 for soft dosage forms. Results of microbiological screening on the reference microorganism test strains showed the highest antimicrobial activity of the ointment composition sample № 8, which contains (wt.%): EXB - 5,0; propylene glycol - 10,0; emulsifier 1 - 5,0; PEO 1500 - 28,0; PEO 400 - 52,0. For the first time it was proved that the new ointment composition with EXB № 8 is superior to the "Calendula ointment" analogue preparation; by its antimicrobial activity with respect to clinical isolates of dominant species of wound infection pathogens: A. baumannii, E. faecalis, K. pneumoniae, P. aeruginosa, C. perfringens, S. aureus, S. epidermidis, S. pyogenes, P. vulgaris, B. subtilis, C. albicans. A new ointment composition based on EXB 5% with excipients with antimicrobial activity is proposed for the next stages of testing the industrial production of the drug with subsequent use in surgical practice to improve the effectiveness of treatment of infected wounds.
Degree of implementation: 1 patent of Ukraine for useful model, 1 guideline, 1 information letter of Ministry of Health of Ukraine, 1 innovation, 4 monographs (3 - foreign), 3 certificates of implementation. Scope (area) of use: health care (medicine).
