Identification of novel small-molecule inhibitors of SARS-CoV-2 by chemical genetics  

作　　者：Chris Chun-Yiu Chan Qian Guo Jasper Fuk-Woo Chan Kaiming Tang Jian-Piao Cai Kenn Ka-Heng Chik Yixin Huang Mei Dai Bo Qin Chon Phin Ong Allen Wing-Ho Chu Wan-Mui Chan Jonathan Daniel Ip Lei Wen Jessica Oi-Ling Tsang Tong-Yun Wang Yubin Xie Zhenzhi Qin Jianli Cao Zi-Wei Ye Hin Chu Kelvin Kai-Wang To Xing-Yi Ge Tao Ni Dong-Yan Jin Sheng Cui Kwok-Yung Yuen Shuofeng Yuan 

机构地区：[1]State Key Laboratory of Emerging Infectious Diseases,Carol Yu Centre for Infection,Department of Microbiology,School of Clinical Medicine,Li Ka Shing Faculty of Medicine,the University of Hong Kong,Pokfulam,Hong Kong SAR 999077,China [2]Department of Infectious Diseases and Microbiology,the University of Hong Kong-Shenzhen Hospital,Shenzhen 518000,China [3]Centre for Virology,Vaccinology and Therapeutics,Hong Kong Science and Technology Park,Hong Kong SAR 999077,China [4]Academician Workstation of Hainan Province,Hainan Medical University-the University of Hong Kong Joint Laboratory of Tropical Infectious Diseases,Haikou 571100,China [5]School of Biomedical Sciences,Li Ka Shing Faculty of Medicine,the University of Hong Kong,Pokfulam,Hong Kong SAR 999077,China [6]NHC Key Laboratory of Systems Biology of Pathogens,Institute of Pathogen Biology Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100730,China [7]College of Biology,Hunan Provincial Key Laboratory of Medical Virology,Hunan University,Changsha 410082,China

出　　处：《Acta Pharmaceutica Sinica B》2024年第9期4028-4044,共17页药学学报（英文版）

基　　金：National Natural Science Foundation of China(NSFC)/Research Grants Council(RGC)Joint Research Scheme(N_HKU767/22 and 82261160398);Health and Medical Research Fund(COVID190121);the Food and Health Bureau,The Government of the Hong Kong Special Administrative Region;the National Natural Science Foundation of China(32322087,32300134,and 82272337);Guangdong Natural Science Foundation(2023A1515012907);Health@-InnoHK,Innovation and Technology Commission,the Government of the Hong Kong Special Administrative Region;the Collaborative Research Fund(C7060-21G and C7002-23Y)and Theme-Based Research Scheme(T11-709/21-N)of the Research Grants Council,The Government of the Hong Kong Special Administrative Region;Partnership Programme of Enhancing Laboratory Surveillance and Investigation of Emerging Infectious Diseases and Antimicrobial Resistance for the Department of Health of the Hong Kong Special Administrative Region Government;Sanming Project of Medicine in Shenzhen,China(SZSM201911014);the High Level-Hospital Program,Health Commission of Guangdong Province,China;the research project of Hainan Academician Innovation Platform(YSPTZX202004);Emergency Collaborative Project of Guangzhou Laboratory(EKPG22-01);and the National Key R&D Program of China(projects 2021YFC0866100 and 2023YFC3041600);The University of Hong Kong Seed Fund for Collaborative Research(2207101537);and Hunan University(521119400156);donations of Providence Foundation Limited(in memory of the late Lui Hac Minh).

摘　　要：There are only eight approved small molecule antiviral drugs for treating COVID-19.Among them,four are nucleotide analogues(remdesivir,JT001,molnupiravir,and azvudine),while the other four are protease inhibitors(nirmatrelvir,ensitrelvir,leritrelvir,and simnotrelvir-ritonavir).Antiviral resistance,unfavourable drug‒drug interaction,and toxicity have been reported in previous studies.Thus there is a dearth of new treatment options for SARS-CoV-2.In this work,a three-tier cell-based screening was employed to identify novel compounds with anti-SARS-CoV-2 activity.One compound,designated 172,demonstrated broad-spectrum antiviral activity against multiple human pathogenic coronaviruses and different SARS-CoV-2 variants of concern.Mechanistic studies validated by reverse genetics showed that compound 172 inhibits the 3-chymotrypsin-like protease(3CLpro)by binding to an allosteric site and reduces 3CLpro dimerization.A drug synergistic checkerboard assay demonstrated that compound 172 can achieve drug synergy with nirmatrelvir in vitro.In vivo studies confirmed the antiviral activity of compound 172 in both Golden Syrian Hamsters and K18 humanized ACE2 mice.Overall,this study identified an alternative druggable site on the SARS-CoV-23CLpro,proposed a potential combination therapy with nirmatrelvir to reduce the risk of antiviral resistance and shed light on the development of allosteric protease inhibitors for treating a range of coronavirus diseases.

关 键 词：SARS-CoV-2 High throughput screening Broad-spectrum antiviral treatment 3CLpro inhibitor Allosteric-site inhibitor Animal models Chemical genetics Reverse genetics 

分 类 号：R392.2[医药卫生—免疫学]