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Herein, the reaction\u2010induced self\u2010assembly (RISA) of polymyxins and natural aldehydes in aqueous solution is presented. The resulting assemblies effectively mask the positively charged nature of polymyxins, reducing their cytotoxicity. Moreover, the representative PMB<jats:bold>A<jats:sub>4<\/jats:sub><\/jats:bold> (composed of polymyxin B (PMB) and (<jats:italic>E<\/jats:italic>)\u20102\u2010heptenal (<jats:bold>A<jats:sub>4<\/jats:sub><\/jats:bold>)) assemblies demonstrate enhanced binding to Gram\u2010negative bacterial outer membranes and exhibit multiple antimicrobial mechanisms, including increased membrane permeability, elevated bacterial metabolism, suppression of quorum sensing, reduced ATP synthesis, and potential reduction of bacterial drug resistance. Remarkably, PMB<jats:bold>A<jats:sub>4<\/jats:sub><\/jats:bold> assemblies reverse drug resistance in clinically isolated drug\u2010resistant strains of Gram\u2010negative bacteria, demonstrating exceptional efficacy in preventing and eradicating bacterial biofilms. PMB<jats:bold>A<jats:sub>4<\/jats:sub><\/jats:bold> assemblies efficiently eradicate Gram\u2010negative bacterial biofilm infections in vivo and alleviate inflammatory response. This RISA strategy offers a practical and clinically applicable approach to minimize side effects, reverse drug resistance, and prevent the emergence of resistance associated with free polymyxins.<\/jats:p>","DOI":"10.1002\/adma.202406156","type":"journal-article","created":{"date-parts":[[2024,7,18]],"date-time":"2024-07-18T05:10:29Z","timestamp":1721279429000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":31,"title":["Reaction\u2010Induced Self\u2010Assembly of Polymyxin Mitigates Cytotoxicity and Reverses Drug Resistance"],"prefix":"10.1002","volume":"36","author":[{"given":"Xiaowen","family":"Hu","sequence":"first","affiliation":[{"name":"Joint Centre of Translational Medicine Key Laboratory of Clinical Laboratory Diagnosis and Translational Research of Zhejiang Province the First Affiliated Hospital of Wenzhou Medical University  Wenzhou Zhejiang 325035 China"},{"name":"Department of Orthodontics School and Hospital of 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