Cupferron impairs the growth and virulence of Escherichia coli clinical isolates.

Abstract

Multidrug resistance is a worrying problem worldwide. The lack of readily available drugs to counter nosocomial infections requires the need for new interventional strategies. Drug repurposing represents a valid alternative to using commercial molecules as antimicrobial agents in a short time and with low costs. Contextually, the present study focused on the antibacterial potential of the ammonium salt N-nitroso-N-phenylhydroxylamine (Cupferron), evaluating the ability to inhibit microbial growth and influence the main virulence factors. Cupferron cytotoxicity was checked via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and hemolysis assays. The antimicrobial activity was assessed through the Kirby-Bauer disk diffusion test, broth microdilution method and time-killing kinetics. Furthermore, the impact on different stages of the biofilm life cycle, catalase, swimming and swarming motility was estimated via MTT and crystal violet (CV) assay, H2O2 sensitivity and motility tests, respectively. Cupferron exhibited less than 15% cytotoxicity at 200 μg/mL concentration. The 90% bacterial growth inhibitory concentrations (MIC90) values recorded after 24 hours of exposure were 200 and 100 μg/mL for multidrug-resistant (MDR) and sensitive strains, respectively, exerting a bacteriostatic action. Cupferron-treated bacteria showed increased susceptibility to biofilm production, oxidative stress and impaired bacterial motility in a dose-dependent manner. In the new antimicrobial compounds active research scenario, the results indicated that Cupferron could be an interesting candidate for tackling E. coli infections.