PaAP-Activatable NIR Probe for Diagnosing, Imaging, and Discovering Small-Molecule Therapeutics against Implant-Associated Biofilm Infections

Biofilm formation on medical implants causes implant-associated infections (IAIs), leading to high morbidity and mortality. Developing molecular tools for precise biofilm detection, along with novel strategies and agents to target biofilm-related IAIs, is crucial for improving treatment options and patient outcomes. Pseudomonas aeruginosa aminopeptidase (PaAP), a key biofilm-associated virulence factor, is a promising target for combating infections. Here, we developed a PaAP-activatable near-infrared (NIR) fluorescent probe, Hcy-NEO-Leu, for real-time, specific, and sensitive detection of PaAP activity. This probe enables noninvasive imaging of the P. aeruginosa biofilm in vitro and in vivo. The probe also identified LY-58, a lycorine derivative that disrupts biofilm formation without affecting bacterial growth or mammalian cell viability, enhancing tobramycin penetration and overcoming antibiotic resistance. This study introduces LY-58 as a promising adjunctive therapy. In conclusion, the PaAP-activatable NIR imaging probe, combined with LY-58, offers innovative tools for the early diagnosis and effective treatment of IAIs.