Identification of Antibiofilm Agents against Salmonella enterica from the Pathogen Box Compound Library

BackgroundBiofilms are central to Salmonella pathogenesis, and targeting their formation is believed to produce less evolutionary pressure of growth inhibition than traditional antibacterials. In this study, we screened the Medicines for Malaria Venture (MMV) Pathogen Box library to identify anti-biofilm agents against S. enterica that possess drug-like properties. Methodology/ Principal FindingsA crystal-violet-based medium-throughput antibiofilm screen of Salmonella enterica serovar Typhimurium ATCC 14028 and a clinical Salmonella enterica serovar Elisabethville isolate was performed on polystyrene surfaces using the 400-compound Pathogen Box library. Compounds that inhibited biofilm formation by >30% and growth by <10% were identified as hits. Salmonella red-dry-rough and motility phenotypes were explored in mechanism of action studies on one hit compound. The Salmonella antibiofilm hit rate was 0.75% for this library. MMV688371 (benzamide) inhibited biofilm formation of S. Typhimurium ATCC 14028 by 33% without inhibiting growth. An ethambutol analogue (MMV687273) and auranofin (MMV688978) met the hit criteria against S. Elisabethville LLD035X. Auranofin showed concentration-dependent, growth-inhibition-independent antibiofilm activity against typhoidal and non-typhoidal Salmonella from Nigeria, and inhibited the motility of S. Elisabethville LLD035X at 5 {micro}M. At 5 {micro}M aurothioglucose, an auranofin gold (I) analogue, and non-gold analogue 1-Thio-beta-D-glucose tetraacetate, inhibited biofilm formation by 61.30% and 11.39%, respectively, pointing to essentiality of the gold (I) moiety for activity. Conclusions/ SignificanceStructurally diverse small molecules can inhibit biofilm formation by Salmonella, and motility inhibition is an important mechanism for this activity. Auranofin inhibits typhoidal and non-typhoidal Salmonella biofilm formation, with its gold content being required for these activities.