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Discovery of the covalent SARS-CoV-2 M pro inhibitors from anti-viral herbs via integrating target-based high-throughput screening and chemoproteomic approaches
  • +8
  • Guang-Bo Ge,
  • Ya-Ni Zhang,
  • Guang-Hao Zhu,
  • Wei Liu,
  • Xi-Xiang Chen,
  • Yuan-Yuan Xie,
  • Jian-Rong Xu,
  • Mei-Fang Jiang,
  • Xiao-yu Zhuang,
  • Wei-Dong Zhang,
  • Hong-Zhuan Chen
Guang-Bo Ge
Shanghai University of Traditional Chinese Medicine

Corresponding Author:geguangbo@shutcm.edu.cn

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Ya-Ni Zhang
Shanghai University of Traditional Chinese Medicine
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Guang-Hao Zhu
Shanghai University of Traditional Chinese Medicine
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Wei Liu
Shuguang Hospital
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Xi-Xiang Chen
Shanghai University of Traditional Chinese Medicine
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Yuan-Yuan Xie
Shanghai University of Traditional Chinese Medicine
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Jian-Rong Xu
Shanghai University of Traditional Chinese Medicine
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Mei-Fang Jiang
Shanghai University of Traditional Chinese Medicine
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Xiao-yu Zhuang
Shanghai University of Traditional Chinese Medicine
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Wei-Dong Zhang
Shanghai University of Traditional Chinese Medicine
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Hong-Zhuan Chen
Shanghai University of Traditional Chinese Medicine
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Abstract

The main proteases (M pro) are highly conserved cysteine-rich proteins that can be covalently modified by numerous natural and synthetic compounds. Herein, we constructed an integrative approach to efficiently discover covalent inhibitors of M pro from complex herbal matrices. This work begins with biological screening of sixty clinically used antiviral herbal medicines, among which Lonicera japonica (LJ) demonstrated the strongest anti-M pro effect (IC 50 = 37.82 μg/mL). Mass spectrometry-based chemical analysis and chemoproteomic profiling revealed that LJ extract contains at least 50 constituents, of which 22 exhibited the capability to covalently modify M pro. We subsequently verified the anti-M pro effects of these covalent binders. Gallic acid and quercetin were found to potently inhibit SARS-CoV-2 M pro in dose- and time- dependent manners, with the IC 50 values below 10 µM. The inactivation kinetics, binding affinity and binding mode of gallic acid and quercetin were further characterized by fluorescence resonance energy transfer, surface plasmon resonance, and covalent docking simulations. Overall, this study established a practical approach for efficiently discovering the covalent inhibitors of M pro from herbal medicines by integrating target-based high-throughput screening and mass spectrometry-based assays, which would greatly facilitate the discovery of key anti-viral constituents from medicinal plants.
15 Jun 2023Submitted to Journal of Medical Virology
11 Jul 2023Submission Checks Completed
11 Jul 2023Assigned to Editor
11 Jul 2023Review(s) Completed, Editorial Evaluation Pending
17 Jul 2023Reviewer(s) Assigned
07 Aug 2023Editorial Decision: Revise Major
21 Sep 20231st Revision Received
21 Sep 2023Submission Checks Completed
21 Sep 2023Assigned to Editor
21 Sep 2023Review(s) Completed, Editorial Evaluation Pending
21 Sep 2023Reviewer(s) Assigned
17 Oct 2023Editorial Decision: Accept