Strategy for cysteine-targeting covalent inhibitors screening using in-house database based LC-MS/MS and drug repurposing

Xiaolan Hu; Jian-Lin Wu; Quan He; Zhi-Qi Xiong; Na Li

doi:10.1016/j.jpha.2024.101045

Volume 15 Issue 3

Apr. 2025

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Xiaolan Hu, Jian-Lin Wu, Quan He, Zhi-Qi Xiong, Na Li. Strategy for cysteine-targeting covalent inhibitors screening using in-house database based LC-MS/MS and drug repurposing[J]. Journal of Pharmaceutical Analysis, 2025, 15(3): 101045. doi: 10.1016/j.jpha.2024.101045

Citation:

Xiaolan Hu, Jian-Lin Wu, Quan He, Zhi-Qi Xiong, Na Li. Strategy for cysteine-targeting covalent inhibitors screening using in-house database based LC-MS/MS and drug repurposing[J]. Journal of Pharmaceutical Analysis, 2025, 15(3): 101045. doi: 10.1016/j.jpha.2024.101045

Citation:

Xiaolan Hu, Jian-Lin Wu, Quan He, Zhi-Qi Xiong, Na Li. Strategy for cysteine-targeting covalent inhibitors screening using in-house database based LC-MS/MS and drug repurposing[J]. Journal of Pharmaceutical Analysis, 2025, 15(3): 101045. doi: 10.1016/j.jpha.2024.101045

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Strategy for cysteine-targeting covalent inhibitors screening using in-house database based LC-MS/MS and drug repurposing

doi: 10.1016/j.jpha.2024.101045

Xiaolan Hu^a,b,
Jian-Lin Wu^{a
,
,},
Quan He^a,
Zhi-Qi Xiong^c,
Na Li^{a
,
,}

a. State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Taipa, Macau SAR, 999078, China;
b. College of Environment and Climate, Institute of Mass Spectrometry and Atmospheric Environment, Guangdong Provincial Key Laboratory of Speed Capability Research, Jinan University, Guangzhou, 510632, China;
c. Center for Excellence in Brain Science and Intelligence Technology, Institute of Neuroscience and State Key Laboratory of Neuroscience, Chinese Academy of Sciences, Shanghai, 200031, China

Funds:

This work was supported by the Science and Technology Development Fund, Macau SAR, China (Grant Nos.: FDCT 0001/2020/AKP and 006/2023/SKL) and Guangxi Science and Technology Major Program, China (Program No.: Guike AA22096022).

Received Date: Apr. 07, 2024
Accepted Date: Jul. 16, 2024
Rev Recd Date: Jul. 14, 2024
Publish Date: Jul. 18, 2024

Abstract

Abstract

Targeted covalent inhibitors, primarily targeting cysteine residues, have attracted great attention as potential drug candidates due to good potency and prolonged duration of action. However, their discovery is challenging. In this research, a database-assisted liquid chromatography-tandem mass spectrometry (LC-MS/MS) strategy was developed to quickly discover potential cysteine-targeting compounds. First, compounds with potential reactive groups were selected and incubated with N-acetyl-cysteine in microsomes. And the precursor ions of possible cysteine-adducts were predicted based on covalent binding mechanisms to establish in-house database. Second, substrate-independent product ions produced from N-acetyl-cysteine moiety were selected. Third, multiple reaction monitoring scan was conducted to achieve sensitive screening for cysteine-targeting compounds. This strategy showed broad applicability, and covalent compounds with diverse structures were screened out, offering structural resources for covalent inhibitors development. Moreover, the screened compounds, norketamine and hydroxynorketamine, could modify synaptic transmission-related proteins in vivo, indicating their potential as covalent inhibitors. This experimental-based screening strategy provides a quick and reliable guidance for the design and discovery of covalent inhibitors.
- Cysteine-targeting inhibitors screening,
- Drug repurposing,
- Metabolites,
- LC-MS/MS,
- Protein targets

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References(49)

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