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Leveraging Protein Dynamics for Selective Inhibition of Threonyl-tRNA Synthetase by Obafluorin Analogs.

Created on 01 Sep 2026

Authors

Zilu Wang, Zhengxuan Zhang, Mingyu Xia, Min Fang, Haoran Xu, Hang Qiao, Wenping Ding, Jiancheng Huang, Guozheng Li, Ke Luo, Wen Liu, Jing Wang, Biao Yu, Pengfei Fang

Published in

Angewandte Chemie (International ed. in English). Pages e5509036. Sep 01, 2026. Epub Sep 01, 2026.

Abstract

The widespread emergence of antibiotic resistance necessitates the development of novel agents with unique mechanisms of action. Obafluorin (OB), a natural β-lactone antibiotic, is a covalent inhibitor of threonyl-tRNA synthetase (ThrRS), but the high conservation of the active site between prokaryote and eukaryote ThrRSs results in minimal selectivity, hindering the therapeutic potential of OB. Here, we report a structure dynamics-based design strategy that transforms OB into a selective antibacterial agent. OB inhibits human and bacterial ThrRSs with nearly equal potency due to identical binding modes. The nitrophenyl moiety of OB is proposed as a 'kinetic sensor' that discriminates between sensitive and resistant ThrRS paralogs. Guided by this insight, we designed a series of OB analogs through rational modification of this moiety. Among them, OB-D4 bearing a para-methoxyphenyl group in place of the nitrophenyl group, exhibited a 241-fold selectivity for bacterial over human ThrRS, along with a markedly improved safety profile with minimal cytotoxicity. In a murine skin infection model, OB-D4 effectively eradicated pathogens, resolving inflammation, and promoting wound healing. Together, this work establishes a 'kinetic sensor' strategy for achieving species selectivity, turning a fundamental challenge in drug discovery-high active-site conservation-into an exploitable opportunity based on dynamic differences.

PMID:
42677972
Bibliographic data and abstract were imported from PubMed on 01 Sep 2026.

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