2. Academic Validation
  3. Discovery of Potent, Allosteric GSTO1 Covalent Inhibitors with a New Binding Mode

Discovery of Potent, Allosteric GSTO1 Covalent Inhibitors with a New Binding Mode

  • J Med Chem. 2025 Nov 27;68(22):24075-24093. doi: 10.1021/acs.jmedchem.5c01724.
Yanli Sun 1 2,  Fengying Zhang 3,  Lixin Zhou 1 4,  Yixuan Feng 1 2 5,  Shuke Yang 1 2,  Qiuxian Lu 3,  Xin Yuan 3,  Wenqing Hou 3,  Biwei Liu 3,  Zhixiang Guo 6,  Lu Chen 1,  Bo Peng 1,  Xianzhen Yin 1,  Yiwen Zhang 5,  Jing Yang 4,  Nan Chen 3,  Wenchao Lu 1 2
Affiliations

Affiliations

  • 1 Lingang Laboratory, Shanghai 200031, China.
  • 2 School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China.
  • 3 ChomiX Biotech Co. Ltd., Nanjing 210000, China.
  • 4 Department of Hematology, Tongji Hospital, Frontier Science Center for Stem Cell Research, Shanghai Key Laboratory of Signaling and Disease Research, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
  • 5 State Key Laboratory of Biotherapy, West China Hospital, Sichuan University and Collaborative Innovation Center for Biotherapy, Chengdu 610041, China.
  • 6 Department of Cardiovascular Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei 230022, China.
PMID: 41233953 DOI: 10.1021/acs.jmedchem.5c01724
Abstract

GSTO1 activates the NLRP3 inflammasome by deglutathionylating NEK7 at Cys253, driving proinflammatory responses. Using PRM-based targeted mass spectrometry, we identified compound A1 as a covalent GSTO1 inhibitor modifying Cys32. Ligand-based optimization generated analogues with diverse activities; among them, A13 exhibited moderate target engagement (kinact/KI = 226 M-1·s-1), excellent cysteine selectivity confirmed by desthiobiotin iodoacetamide (DBIA)- and alkyne-based ABPP, and superior metabolic stability in human liver microsomes. High-resolution crystal structures revealed an unexpected A13 binding mode occupying a new hydrophobic pocket distinct from known GSTO1 inhibitors. Functionally, covalent targeting of GSTO1-C32 by A13 markedly reduced LPS-induced IL-1β and IL-18 secretion in human monocyte-derived macrophages. Collectively, these results identify A13 as a potent, selective, and metabolically stable lead compound for developing next-generation GSTO1 inhibitors targeting inflammatory diseases.

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