Boaro, AndreiaAgeitos, LuciaTorres, Marcelo Der TorossianBlasco, Esther BrosetOztekin, Sebahatde la Fuente-Nunez, Cesar2024-10-042024-10-0420232666-3864https://doi.org/10.1016/j.xcrp.2023.101459http://hdl.handle.net/20.500.12403/3708Antimicrobial peptides (AMPs) derived from natural toxins and venoms offer a promising alternative source of antibiotics. Here, through structure-function-guided design, we convert two natural AMPs derived from the venom of the solitary eumenine wasp Eu-menes micado into a-helical AMPs with reduced toxicity that kill Gram-negative bacteria in vitro and in a preclinical mouse model. To identify the sequence determinants conferring antimicrobial ac-tivity, an alanine scan screen and strategic single lysine substitutions are made to the amino acid sequence of these natural peptides. These efforts yield a total of 34 synthetic derivatives, including alanine substituted and lysine-substituted sequences with stabilized a-helical structures and increased net positive charge. The resulting lead synthetic peptides kill the Gram-negative pathogens Escheri-chia coli and Pseudomonas aeruginosa (PAO1 and PA14) by rapidly permeabilizing both their outer and cytoplasmic membranes, exhibit anti-infective efficacy in a mouse model by reducing bacte-rial loads by up to three orders of magnitude, and do not readily select for bacterial resistance.eninfo:eu-repo/semantics/openAccessCationic Antimicrobial PeptidesPseudomonas-AeruginosaPolycationic PeptidesResistanceBacterialMembraneAntibioticsMechanismSynergyAnalogsArticle4710.1016/j.xcrp.2023.10145938239869WOS:001047018200001Q1