Overcoming Clinical Barriers: Utilizing Self‐Assembly Systems to Break the Protease Limitations of Antimicrobial Peptides

ABSTRACT 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 (GM MIC = 2.88) and the greatest clinical potential (GM SI = 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.