Drug screening to identify compounds to eliminate Burkholderia pseudomallei through Hcp protein.

Burkholderia pseudomallei (B. pseudomallei) exhibits inherent resistance to multiple antibiotics and Strong pathogenicity, highlighting the urgent need for effective antibiotics. The type VI secretion system (T6SS) is a key virulence factor in B. pseudomallei, and its core structural protein hemolysin coregulated protein (Hcp) represents a conserved, essential target for potential intervention. In this study, Hcp was utilized as a molecular target to screen antibacterial agents from the FDA Orange Book database. Subsequent functional assays and mechanistic analyses identified netilmicin sulfate, which specifically binds to Hcp and exhibits a potent bacteriostatic effect. In vitro and in vivo studies demonstrated that it effectively eliminates B. pseudomallei and enhances host survival post-infection. Combination with ceftazidime exhibits synergism, strengthening host cell protection and reducing the required dosage. Mechanistically, it disrupts Hcp's tubular structure, compromising T6SS functional architecture and attenuating B. pseudomallei viability. Beyond identifying a promising antibiotic candidate, this approach underscores a potential strategy for future antimicrobial drug discovery targeting T6SS components.