iProXProteomicsRui WangEscherichia Colihttp://www.iprox.org/page/project.html?id=IPX0012231000wangrui@szbl.ac.cnshenzhen bay laboratoryiProXfalseRational Design of Benzo-Dioxygenated FabH Inhibitors Unveiled by Proteomic Profiling: Broad-Spectrum Antibacterial Activity and In Vivo Therapeutic EfficacyThe global escalation of antibiotic-resistant bacterial infections poses a life-threatening challenge to public health, necessitating the urgent development of innovative antibiotics targeting unexploited metabolic vulnerabilities. Through rational design, we developed a series of benzo-dioxygenated FabH inhibitors targeting the bacterial fatty acid biosynthesis pathway. Guided by active-site analysis and pharmacophore-guided optimization, we engineered a Y-shaped scaffold that achieved nanomolar inhibition of FabH (IC50 = 1.90 µM). The lead compound F35 showed broad-spectrum efficacy with MIC values as low as 1.56 µg/mL against Gram-negative and Gram-positive pathogens, outperforming Kanamycin B. Structural analysis revealed key interactions between FabH conserved residues and fluorine-mediated halogen bonding. In vivo assay, F35 accelerated wound closure in S. aureus-infected rodents, demonstrating a favorable biocompatibility. Our study establishes a convergence paradigm that integrates structure design, chemoproteomic identification, and therapeutic development for antibiotics, providing a strategic blueprint to combat multidrug resistance via precision targeting of metabolic chokepoints in bacterial pathogens.Tue Jun 10 00:00:00 GMT+01:00 2025PXD064801562