Cross‐Strand Interaction, Central Bending, and Sequence Pattern Act as Biomodulators of Simplified β‐Hairpin Antimicrobial Amphiphiles

Abstract Novel antimicrobial peptides (AMPs) have revolutionarily evolved into formidable candidates for antibiotic substitute materials against pathogenic infections. However, cost, lability, disorderly sequences, systemic toxicology, and biological profiles have plagued the perennial search. Here, a progressive β‐hairpin solution with the simplest formulation is implanted into an AMP‐based therapeutic strategy to systematically reveal the complex balance between function and toxicity of structural moieties, including cationicity, hydrophobicity, cross‐strand interactions, center bending, and sequence pattern. Comprehensive implementation of structural identification, ten microorganisms, eleven in vitro barriers, four mammalian cells, and a diversified membrane operation setup led to the emergence of β‐hairpin prototypes from a 24‐member library. Lead amphiphiles, WKF‐PG and WRF‐NG, can tackle bacterial infection through direct antimicrobial efficacy and potential inflammation‐limiting capabilities, such as an Escherichia coli challenge in a mouse peritonitis‐sepsis model, without observed toxicity after systemic administration. Their optimal states with dissimilar modulators and the unavailable drug resistance related to membrane lytic mechanisms, also provide an usher for renewed innovation among β‐sheet peptide‐based antimicrobial biomaterials.