2. Academic Validation
  3. Optimized ebselen derivatives as novel potent Escherichia coli β-glucuronidase covalent allosteric inhibitors

Optimized ebselen derivatives as novel potent Escherichia coli β-glucuronidase covalent allosteric inhibitors

  • Eur J Med Chem. 2025 Jun 5:290:117571. doi: 10.1016/j.ejmech.2025.117571.
Ti-Ti Ying 1,  Hao-Qiang Hu 2,  Xiao-Wen Wu 1,  Xu-Liang Xu 1,  Jian Lv 2,  Shu-Ning Zhang 3,  Hong Wang 4,  Wei Hou 5,  Bin Wei 6,  Guo-Wu Rao 7
Affiliations

Affiliations

  • 1 College of Pharmaceutical Science & Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang Key Laboratory of Green, Low-carbon, and Efficient Development of Marine Fishery Resources, Zhejiang University of Technology, Hangzhou, 310014, China.
  • 2 Institute of Drug Development & Chemical Biology, College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310014, China.
  • 3 Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai, 201210, China.
  • 4 College of Pharmaceutical Science & Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang Key Laboratory of Green, Low-carbon, and Efficient Development of Marine Fishery Resources, Zhejiang University of Technology, Hangzhou, 310014, China; Binjiang Institute of Artificial Intelligence, ZJUT, Hangzhou, 310051, China.
  • 5 Institute of Drug Development & Chemical Biology, College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310014, China. Electronic address: houwei@zjut.edu.cn.
  • 6 College of Pharmaceutical Science & Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang Key Laboratory of Green, Low-carbon, and Efficient Development of Marine Fishery Resources, Zhejiang University of Technology, Hangzhou, 310014, China. Electronic address: binwei@zjut.edu.cn.
  • 7 Institute of Drug Development & Chemical Biology, College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310014, China. Electronic address: rgw@zjut.edu.cn.
PMID: 40168911 DOI: 10.1016/j.ejmech.2025.117571
Abstract

Gut microbial β-glucuronidase (GUS) plays a key role in metabolizing compounds and influencing disease and drug metabolism, highlighting the need for potent inhibitors to improve drug efficacy and intestinal health. To identify Escherichia coli β-glucuronidase (EcGUS) inhibitors, we designed and synthesized fifty 1,2-benzoselenazol-3-one (BSEA) derivatives using a bioisosterism strategy. Among these, twenty-five BSEA derivatives demonstrated greater inhibitory efficacy than the most potent known EcGUS inhibitor, amoxapine (AMX), with compound 49 showing the strongest activity, achieving an IC50 of 12.9 nM. Structure-inhibitory activity relationship analysis suggested that modifications such as adding benzene rings or nitrogenous heterocycles to the BSEA scaffold enhanced inhibitory activity, influenced by the type and position of substituents. The LC-MS analysis confirmed that compounds 31 and 49 covalently modify Cys197 in EcGUS, and additional covalent linkage of compound 49 was observed on Cys28 and Cys443. In addition, the jump dilution assays proved that compounds 31 was irreversible covalent inhibitors, and its kinetic parameter kinact/KI were determined to be 21292.9 M-1s-1. The compounds 49 was reversible covalent inhibitors and its apparent steady-state inhibition constant Ki∗app were determined to be 23.33 nM. Molecular docking predicted specific interactions, such as hydrogen bonds involving Se and the pyrazole NH of compound 49 with Cys28 and Cys449, which may contribute to its inhibitory action. This study reports the first discovery of covalent inhibitors for EcGUS, with optimized BSEA derivatives acting as novel allosteric covalent inhibitors, revealing structure-activity relationships and molecular determinants that establish their potential in drug development.

Keywords

1,2-Benzoselenazol-3-one derivatives; Covalent inhibitor; Gut microbiota; Structure-inhibitory activity relationships; β-glucuronidase.

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