Design of TAT-Conjugated Bowman-Birk Trypsin Inhibitor Peptides with Enhanced Antimicrobial and Antiproliferative...

Cell-penetrating peptide (CPP) conjugation represents a promising strategy for enhancing the biological activity of therapeutic peptides. In this study, three analogues were designed by conjugating the trypsin inhibitory loop (TIL) derived from a Bowman-Birk-type inhibitor with the transactivator of transcription (TAT) peptide to improve their bioactivity. All TAT-TIL conjugates exhibited significantly enhanced antimicrobial activity compared with the parent peptide. Notably, the analogue containing a glycine linker (-GG-) showed further improvement in antiproliferative activity against cancer cells, indicating the potential role of linker design in optimizing peptide function. All analogues exhibited low hemolytic activity at the highest tested concentrations, although increased cytotoxicity toward normal HaCaT cells was observed, suggesting the need for further optimization of selectivity. Interestingly, comparable antimicrobial activities were observed regardless of protease inhibitory capacity, indicating that protease inhibition is not essential for the enhanced biological effects. Overall, TAT conjugation significantly improves the biological activity of Bowman-Birk-type inhibitor-derived peptides, and the incorporation of a glycine linker further enhances their functional properties. These findings support CPP-mediated peptide modification as an effective strategy for developing potential antimicrobial and anticancer peptide candidates.