摘要Given the increasing concern regarding antibacterial resistance,the antimicrobial properties of naphthoquinones have recently attracted significant attention.While 1,4-naphthoquinone and its derivatives have been extensively studied,the antibacterial properties of 5,8-di-hydroxy-1,4-naphthoquinone derivatives remain relatively unexplored.This study presents a comprehensive in vitro and in vivo analysis of the antibacterial activity of 35 naturally sourced and chemically synthesized derivatives of 5,8-dihydroxy-1,4-naphthoquinone.Kirby-Bauer antibiotic testing identified three compounds with activity against methicillin-resistant Sta-phylococcus aureus(MRSA),with one compound(PNP-02)demonstrating activity compar-able to vancomycin in minimum inhibitory concentration,minimum bactericidal concentra-tion(MBC),and time-kill assays.Microscopic and biochemical analyses revealed that PNP-02 adversely affects the cell wall and cell membrane of MRSA.Mechanistic investigations,includ-ing proteomic sequencing analyses,Western blotting,and RT-qPCR assays,indicated that PNP-02 compromises cell membrane integrity by inhibiting arginine biosynthesis and pyrimidine metabolism pathways,thereby increasing membrane permeability and inducing bacterial death.In an in vivo mouse model of skin wound healing,PNP-02 exhibited antibacterial effic-acy similar to vancomycin.The compound demonstrated low toxicity to cultured human cells and in hemolysis assays and remained stable during serum incubation.These findings sug-gest that PNP-02 possesses promising bioactivity against MRSA and represents a potential novel antibacterial agent.