Natural antimicrobial peptides (AMPs) encounter significant challenges in transitioning to clinical application, primarily due to low bioactivity, high toxicity, and poor stability. This study proposes a strategy to enhance the stability of AMPs through molecular assembly while exploring the advantages of the newly designed self-assembled peptides compared to unimer peptides. We conducted a comprehensive investigation of antimicrobial activity, biocompatibility, in vitro stability, and particularly protease stability, aiming to develop highly efficient and stable designer peptides as alternatives to traditional antibiotics. A series of designer peptides with self-assembling capabilities was constructed by attaching various hydrophobic scaffolds to an enzyme-resistant short peptide sequence. The self-assembled designer peptide Pba* with 1-pyrenebutyric acid (Pba) as the hydrophobic scaffold exhibited the highest antibacterial activity (GMMIC = 2.88) and the greatest clinical potential (GMSI = 44.44), while maintaining excellent biocompatibility and physiological stability. Mechanistic studies revealed that Pba* self-assembled into spherical micelles and nanofibers, trapping bacteria and disrupting cell membranes, interfering with respiration and energy metabolism. Notably, Pba* displayed negligible toxicity and alleviated bacterial infections in mice. This study paves the way for the development of highly effective antimicrobial materials.
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