Computer-Aided Rational Modification of Hst 5 Based on Ssa1/2 and the Antifungal Activity of the Derivatives against...

The natural antifungal peptide Histatin 5 (Hst 5) is a histidine-rich cationic peptide secreted by human salivary glands and a key component of oral innate immunity, but its moderate activity limits clinical use. Hst 5 enters Candida albicans via the membrane receptor Ssa1/2. Here, we integrated artificial intelligence-assisted and computer-aided drug design to rationally modified the sequence structure of Hst 5. Truncated derivatives of Hst5 were screened for antimicrobial potential using ESM2-AFPpred, and high-probability candidates were docked with Ssa1/2. The Hst 5-22 was identified, then redesigned based on alanine scanning to yield the optimized derivative Hst 5-22-RW. Compared with Hst 5, Hst 5-22-RW has a shorter sequence, stronger Ssa1/2 binding, and improved activity against C. albicans. It also shows superior activity against fluconazole-resistant strains. RT-qPCR and transmembrane tracking confirmed higher cellular transport efficiency in C. albicans. The CADD/AIDD-driven optimization successfully generated the highly active antifungal peptide Hst 5-22-RW, providing a novel strategy for rational modification of antimicrobial peptides.