Molina Panadero I.Moreno Rodríguez A.Rey Hidalgo A.de la Cruz M.Sánchez P.Tomás Gallardo L.Samernate T.Sencanski M.Glisic S.Genilloud O.Nonejuie P.Pérez-Pulido A.J.Hmadcha A.Smani Y.Mahidol University2025-11-072025-11-072025-01-01EMBO Molecular Medicine (2025)17574676https://repository.li.mahidol.ac.th/handle/123456789/112947High-throughput screening studies provide an additional approach to discovering repurposed drugs for antimicrobial treatments. In this work, we report the identification of ENOblock, an anticancer drug, as an antimicrobial agent. We computationally and experimentally validated that ENOblock synergizes with colistin, the last resort antibiotic. Additionally, we identified enolase as the potential bacterial target for ENOblock. The in silico and in vitro antibacterial activity of ENOblock translated into potent in vivo efficacy in an animal infection model. Collectively, the preclinical data support the selection of ENOblock as a promising candidate for antimicrobial development, with the potential to address the urgent threat of infections caused by Acinetobacter baumannii.Biochemistry, Genetics and Molecular BiologyArticleSCOPUS10.1038/s44321-025-00331-22-s2.0-10502031333917574684