Discovery of Potent, Allosteric GSTO1 Covalent Inhibitors with a New Binding Mode
GSTO1 activates the NLRP3 inflammasome by deglutathionylating NEK7 at Cys253, driving proinflammatory responses. Using PRM-based targeted mass spectrometry, we identified compound <b>A1</b> as a covalent GSTO1 inhibitor modifying Cys32. Ligand-based optimization generated analogues with diverse activities; among them, <b>A13</b> exhibited moderate target engagement (<i>k</i><sub>inact</sub>/<i>K</i><sub>I</sub> = 226 M<sup>-1</sup>·s<sup>-1</sup>), 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 <b>A13</b> binding mode occupying a new hydrophobic pocket distinct from known GSTO1 inhibitors. Functionally, covalent targeting of GSTO1-C32 by <b>A13</b> markedly reduced LPS-induced IL-1β and IL-18 secretion in human monocyte-derived macrophages. Collectively, these results identify <b>A13</b> as a potent, selective, and metabolically stable lead compound for developing next-generation GSTO1 inhibitors targeting inflammatory diseases.
Authors
- Wenchao Lu (ORCID: https://orcid.org/0000-0003-1175-365X)
- Biwei Liu
- Wenqing Hou (ORCID: https://orcid.org/0000-0001-9294-9106)
- Xianzhen Yin (ORCID: https://orcid.org/0000-0002-7870-1718)
- Lixin Zhou (ORCID: https://orcid.org/0000-0002-9362-8117)
- Yixuan Feng (ORCID: https://orcid.org/0000-0001-6596-524X)
- Jing Yang (ORCID: https://orcid.org/0000-0002-3833-9996)
- Yiwen Zhang (ORCID: https://orcid.org/0000-0002-4024-4881)
- Lu Chen (ORCID: https://orcid.org/0000-0002-5685-7017)
- Zhixiang Guo (ORCID: https://orcid.org/0009-0006-1987-2660)
- Fengying Zhang (ORCID: https://orcid.org/0000-0002-8246-1518)
- Yanli Sun (ORCID: https://orcid.org/0000-0002-8883-5226)
- Bo Peng (ORCID: https://orcid.org/0000-0001-9411-716X)
- Xin Yuan (ORCID: https://orcid.org/0000-0001-8907-3123)
- Nan Chen (ORCID: https://orcid.org/0000-0002-4816-6910)
- Qiuyuan Lu (ORCID: https://orcid.org/0009-0007-7382-8297)
- Shuke Yang
Institutions
- Tongji University (CN)
- Anhui Medical University (CN)
- Sichuan University (CN)
- ShanghaiTech University (CN)
- 490 BioTech (United States) (US)
- First Affiliated Hospital of Anhui Medical University (CN)
- Viva Biotech (China) (CN)
- Tongji Hospital (CN)
- State Key Laboratory of Biotherapy
Publication Details
- Journal
- Journal of Medicinal Chemistry
- Published
- 2025-11-14
- DOI
- https://doi.org/10.1021/acs.jmedchem.5c01724
- Citations
- 1
- Primary Topic
- Glutathione Transferases and Polymorphisms
- Type
- article
- Field-Weighted Citation Impact
- 0.81