Computational\nDesign\nof Pore-Forming Peptides with\nPotent Antimicrobial and Anticancer Activities

Peptides that form\ntransmembrane barrel-stave pores are potential\nalternative therapeutics for bacterial infections and cancer. However,\ntheir optimization for clinical translation is hampered by a lack\nof sequence-function understanding. Recently, we have <i>de novo</i> designed the first synthetic barrel-stave pore-forming antimicrobial\npeptide with an identified function of all residues. Here, we systematically\nmutate the peptide to improve pore-forming ability in anticipation\nof enhanced activity. Using computer simulations, supported by liposome\nleakage and atomic force microscopy experiments, we find that pore-forming\nability, while critical, is not the limiting factor for improving\nactivity in the submicromolar range. Affinity for bacterial and cancer\ncell membranes needs to be optimized simultaneously. Optimized peptides\nmore effectively killed antibiotic-resistant ESKAPEE bacteria at submicromolar\nconcentrations, showing low cytotoxicity to human cells and skin model.\nPeptides showed systemic anti-infective activity in a preclinical\nmouse model of Acinetobacter baumannii infection. We also demonstrate peptide optimization for pH-dependent\nantimicrobial and anticancer activity.