Identification of FDA-Approved Drugs as Antivirulence Agents Targeting the <i>pqs</i> Quorum-Sensing System of Pseudomonas aeruginosa

Abstract

The long-term use of antibiotics has led to the emergence of multidrug-resistant bacteria. A promising strategy to combat bacterial infections aims at hampering their adaptability to the host environment without affecting growth. In this context, the intercellular communication system quorum sensing (QS), which controls virulence factor production and biofilm formation in diverse human pathogens, is considered an ideal target. Here, we describe the identification of new inhibitors of the <i>pqs</i> QS system of the human pathogen <i>Pseudomonas aeruginosa</i> by screening a library of 1,600 U.S. Food and Drug Administration-approved drugs. Phenotypic characterization of <i>ad hoc</i> engineered strains and <i>in silico</i> molecular docking demonstrated that the antifungal drugs clotrimazole and miconazole, as well as an antibacterial compound active against Gram-positive pathogens, clofoctol, inhibit the <i>pqs</i> system, probably by targeting the transcriptional regulator PqsR. The most active inhibitor, clofoctol, specifically inhibited the expression of <i>pqs</i>-controlled virulence traits in <i>P. aeruginosa</i>, such as pyocyanin production, swarming motility, biofilm formation, and expression of genes involved in siderophore production. Moreover, clofoctol protected <i>Galleria mellonella</i> larvae from <i>P. aeruginosa</i> infection and inhibited the <i>pqs</i> QS system in <i>P. aeruginosa</i> isolates from cystic fibrosis patients. Notably, clofoctol is already approved for clinical treatment of pulmonary infections caused by Gram-positive bacterial pathogens; hence, this drug has considerable clinical potential as an antivirulence agent for the treatment of <i>P. aeruginosa</i> lung infections.