This work is devoted to the investigation of biocidal properties of quaternary ammonium compounds (QACs) based on pyridine structures with aromatic spacers, and their widely known analogs, against clinically significant microorganisms. This study is focused on investigating their antimicrobial activity (minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs)), antibiofilm properties (minimum biofilm inhibitory concentrations (MBICs) and minimum biofilm eradication concentrations (MBECs)), synergetic effect with different alcohols in antiseptic formulations, and bacterial resistance development. It was shown that all combined analogue preparations had a higher level of antibacterial activity against the tested bacterial strains, with a 16- to 32-fold reduction in MICs and MBCs compared to previously used antiseptic preparations. Moreover, hit-QACs demonstrated a stable effect against Gram-negative E. coli, K. pneumoniae, and A. baumannii within a month of incubation. Overall results indicated a high level of antibacterial activity of pyridine-based QACs.

The increasing antibiotic resistance is a clinical problem worldwide. Numerous Gram-negative bacteria have already become resistant to the most widely used class of antibacterial drugs, β-lactams. One of the main mechanisms is inactivation of β-lactam antibiotics by bacterial β-lactamases. Appearance and spread of these enzymes represent a continuous challenge for the clinical treatment of infections and for the design of new antibiotics and inhibitors. Drug repurposing is a prospective approach for finding new targets for drugs already approved for use. We describe here the inhibitory potency of known detoxifying antidote 2,3-dimercaptopropane-1-sulfonate (unithiol) against metallo-β-lactamases. Unithiol acts as a competitive inhibitor of meropenem hydrolysis by recombinant metallo-β-lactamase NDM-1 with the K_I of 16.7 µM. It is an order of magnitude lower than the K_I for l-captopril, the inhibitor of angiotensin-converting enzyme approved as a drug for the treatment of hypertension. Phenotypic methods demonstrate that the unithiol inhibits natural metallo-β-lactamases NDM-1 and VIM-2 produced by carbapenem-resistant K. pneumoniae and P. aeruginosa bacterial strains. The 3D full atom structures of unithiol complexes with NDM-1 and VIM-2 are obtained using QM/MM modeling. The thiol group is located between zinc cations of the active site occupying the same place as the catalytic hydroxide anion in the enzyme–substrate complex. The sulfate group forms both a coordination bond with a zinc cation and hydrogen bonds with the positively charged residue, lysine or arginine, responsible for proper orientation of antibiotics upon binding to the active site prior to hydrolysis. Thus, we demonstrate both experimentally and theoretically that the unithiol is a prospective competitive inhibitor of metallo-β-lactamases and it can be utilized in complex therapy together with the known β-lactam antibiotics.

The invention and innovation of highly effective antimicrobials are always crucial tasks for medical and organic chemistry, especially at the current time, when there is a serious threat of shortages of effective antimicrobials following the pandemic. In the study presented in this article, we established a new approach to synthesizing three novel series of bioactive water-soluble tris-quaternary ammonium compounds using an optimized one-pot method, and we assessed their antimicrobial and antibiofilm potential. Five pathogenic microorganisms of the ESKAPE group, including highly resistant clinical isolates, were used as the test samples. Moreover, we highlighted the dependence of antibacterial activity from the hydrophilic–hydrophobic balance of the QACs and noted the significant performance of the desired products on biofilms with MBEC as low as 16 mg/L against bacteria and 8 mg/L against fungi. Particularly notable was the high activity against Pseudomonas aeruginosa and Acinetobacter baumannii, which are among the most resilient bacteria known. The presented work will provide useful insights for future research on the topic.

While the pandemic is behind us, the world community faces a global threat of bacterial resistance outbreak. One of the key ways to combat the spread of multi-resistant bacteria is infection prevention and control tactics using modern antiseptic and disinfectant compositions. Herein, we continue the path to unravel the structure–activity relationship (SAR) of potent pyridine-derived biocide class bis-quaternary ammonium compounds (QACs). In this study, twenty dihydroxynaphthalene-derivative bis-QACs were subjected to extensive microbiological analysis on planktonic cells and biofilms of the ESKAPE microorganisms. Among them, hit compounds were superior in their bacteriostatic and bactericidal action to commercial mono-QACs and were comparable to the best bis-QAC antiseptic on the market. SAR analysis indicated that the linker conformation does not significantly affect the activity, though structure symmetry and especially lipophilicity had an influence on antibacterial performance. Furthermore, we delve deeper in investigation of the antimicrobial potential of bis-QACs and conducted a variety of assays, including time–kill kinetics, bacterial resistance formation, cell morphology, and cytotoxicity. Studies showed promising results for compounds 5d and 6d, indicating 2 to 3-fold less cytotoxicity and hemotoxicity compared to commercial QACs. Moreover, SEM imaging revealed that bis-QACs can cause severe membrane damage to S. aureus and P. aeruginosa strains, confirming great potential of novel compounds as antiseptic and disinfectant.

The in vitro antibacterial activity of 11 commercial disinfectant preparations and 8 antiseptics against 10 strains of the bacteria Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterobacter cloaceae and Providencia stuartii obtained from international collections and isolated from neuroresuscitation patients in Moscow in 2018 was studied. The sensitivity of planktonic cultures to the preparations was determined by the method of serial dilutions in broth and the spot method on solid nutrient media, the sensitivity of biofilms by the applicator method. A general pattern was revealed: the level of sensitivity to tested disinfectants in clinical strains was lower than in reference strains. It was found that the disinfectants «Mikrobak-Forte», «SAT-22», «Neobak-Oksi» at the concentrations recommended by the manufacturers were effective against bacteria of all test strains, both in the plankton state and in the form of biofilms. On the contrary, the disinfectant preparations «Biodez-Optima», «Biodez-Extra DVU», «Novodez-Aktiv», «Triosept-Oksi», «Tristel Fusion for Surfaces», «Effect-Forte Plus», «Lactic-Oxy» did not have sufficient effectiveness in the concentrations recommended by the manufacturers, therefore it is proposed to use these drugs in higher concentrations. It was found that the disinfectant «Biodez-Extra DVU» is able to inhibit the growth of biofilms of bacteria of the species K. pneumoniae. The ability to suppress the growth of bacterial biofilms of K. pneumoniae, A. baumannii, P. aeruginosa was revealed for the «Triestel Fusion for surfaces disinfectant». The bacteria of all used test strains in the planktonic state were sensitive to all tested antiseptic preparations. However, the biofilms of the clinical strains of P. aeruginosa and P. stuartii. possessed resistance to the antiseptics «Octenidol», «Octenisept», «Miramistin», «Hexoral». Our studies indicate the need for sensitivity analysis of antibacterial drugs in representatives of hospital pathogens, including the modeling of bacterial biofilms, which is a very relevant and important scientific direction, necessary to improve the control of nosocomial infections in the Russian Federation.

Introduction. With an increase in the level of acquired antibiotic resistance of pathogens, treatment becomes more complicated and slows down, especially in infections associated with biofilms. There is a growing need for the development and use of new antibacterial drugs with specific antimicrobial activity.Aim. To study the antimicrobial action and the dynamics of the formation of resistance to benzydamine hydrochloride from a various infection agents. Materials and methods. To obtain biofilms, microorganisms were cultivated in flat-bottomed culture plates. Planktonic cells were obtained by suspending and reseeding single colonies of the daily culture into flat-bottomed culture plates. To determine the antimicrobial activity of the studied preparations, two-fold dilutions were prepared and added to the wells of the plate with a bacterial culture. The dynamics of the formation of resistance to benzydamine hydrochloride was studied by passaging the cultures in a liquid nutrient medium with increasing concentrations of the antiseptic by a twofold step. After 2–3 days of incubation from a test tube with the maximum concentration of the drug, in which bacterial growth was observed, the bacteria were transferred to new ones with higher concentrations of the drug.Results. It was shown that benzydamine hydrochloride showed a high level of activity against bacteria M. catarrhalis and yeast-like fungi C. albicans. A slightly lower activity of the drug was noted for bacteria of the species S. aureus and E. coli, however, within the limits of the therapeutic concentration of the drug in finished dosage forms. Benzydamine hydrochloride had a significantly higher level of antibacterial activity against pre-formed biofilms compared to drugs such as chlorhexidine and hexetidine. An analysis of the dynamics of the formation of resistance to the drug benzydamine hydrochloride in microorganisms of various species showed that the possibility of developing resistance to benzydamine hydrochloride is extremely small. The process of adaptation was observed only in E. coli. The studied strains of the species S. aureus, C. albicans, and M. catarrhalis did not acquire resistance to the test drug.Conclusion. Benzydamine hydrochloride can be effectively used against a wide range of pathogens of ENT infections, as it has been shown to have a significantly higher level of antibacterial activity against pre-formed biofilms, various types of bacteria and yeast-like fungi and an extremely low level of resistance compared to other antiseptic drugs.