Discovery and characterization of naturally occurring covalent inhibitors of SARS-CoV-2 M^(pro)from the antiviral herb Ephedra  

作　　者：HU Qing ZHANG Yiwen CHEN Pengcheng ZHANG Yani ZHU Guanghao LIU Wei WANG Chaoran ZHENG Shuilian SHEN Nonger WANG Haonan HUANG Ping GE Guangbo 

机构地区：[1]Center for Clinical Pharmacy,Cancer Center,Department of Pharmacy,Zhejiang Provincial People’s Hospital(Affiliated People’s Hospital),Hangzhou Medical College,Hangzhou 310014,China [2]Shanghai Frontiers Science Center of TCM Chemical Biology,Institute of Interdisciplinary Integrative Medicine Research,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China [3]Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200021,China [4]Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116023,China

出　　处：《Chinese Journal of Natural Medicines》2024年第9期797-807,共11页中国天然药物（英文版）

基　　金：supported by the National Key Research and Development Program of China(No.2022YFC-3502000);the Basic Public Welfare Research Program of Zhejiang Province(No.LGF22H280012);Zhejiang Provincial TCM Science and Technology Plan Project(Nos.2023ZR064,GZY-ZJ-KJ-24004,2024ZL007 and 2022ZB017);the Medical Science and Technology Project of Zhejiang Province(Nos.2022495401,2021KY040 and2022KY069);Zhejiang Provincial Key Projects in Chinese Medicine(Nos.2020ZZ003 and 2021ZZ001);Shanghai Science and Technology Innovation Action Plans(Nos.21S21900600);Zhejiang Province"Ten Thousand People Plan"Science and Technology Innovation Leading Talents Project(No.2020R52029)。

摘　　要：The Chinese herb Ephedra(also known as Mahuang)has been extensively utilized for the prevention and treatment of coronavirus-induced diseases,including coronavirus disease 2019(COVID-19).However,the specific anti-SARS-CoV-2 compounds and mechanisms have not been fully elucidated.The main protease(M^(pro))of SARS-CoV-2 is a highly conserved enzyme responsible for proteolytic processing during the viral life cycle,making it a critical target for the development of antiviral therapies.This study aimed to identify naturally occurring covalent inhibitors of SARS-CoV-2 M^(pro)from Ephedra and to investigate their covalent binding sites.The results demonstrated that the non-alkaloid fraction of Ephedra(ENA)exhibited a potent inhibitory effect against the SARS-CoV-2 M^(pro)effect,whereas the alkaloid fraction did not.Subsequently,the chemical constituents in ENA were identified,and the major constituents'anti-SARS-CoV-2 M^(pro)effects were evaluated.Among the tested constituents,herbacetin(HE)and gallic acid(GA)were found to inhibit SARS-CoV-2 M^(pro)in a time-and dose-dependent manner.Their combination displayed a significant synergistic effect on this key enzyme.Additionally,various techniques,including inhibition kinetic assays,chemoproteomic methods,and molecular dynamics simulations,were employed to further elucidate the synergistic anti-M^(pro)mechanisms of the combination of HE and GA.Overall,this study deciphers the naturally occurring covalent inhibitors of SARS-CoV-2 M^(pro)from Ephedra and characterizes their synergistic anti-M^(pro)synergistic effect,providing robust evidence to support the anti-coronavirus efficacy of Ephedra.

关 键 词：Main protease EPHEDRA Ephedra non-alkaloid fraction Covalent inhibitors Synergistic effect 

分 类 号：R965[医药卫生—药理学]