Strategy for cysteine-targeting covalent inhibitors screening using in-house database based LC-MS/MS and drug repurposing
10.1016/j.jpha.2024.101045
- Author:
Xiaolan HU
1
;
Jian-Lin WU
;
Quan HE
;
Zhi-Qi XIONG
;
Na LI
Author Information
1. State Key Laboratory of Quality Research in Chinese Medicine,Macau University of Science and Technology,Taipa,Macau SAR,999078,China;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
- Publication Type:Journal Article
- Keywords:
Cysteine-targeting inhibitors screening;
Drug repurposing;
Metabolites;
LC-MS/MS;
Protein targets
- From:
Journal of Pharmaceutical Analysis
2025;15(3):637-650
- CountryChina
- Language:English
-
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 dis-covery is challenging.In this research,a database-assisted liquid chromatography-tandem mass spec-trometry(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,norket-amine 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.
