High-throughput screening identifies inhibitors of the SARS coronavirus main proteinase

被引:171
作者
Blanchard, JE
Elowe, NH
Huitema, C
Fortin, PD
Cechetto, JD
Eltis, LD [1 ]
Brown, ED
机构
[1] McMaster Univ, Dept Biochem, McMaster High Throughput Screening Lab, Hamilton, ON L8N 3Z5, Canada
[2] Univ British Columbia, Dept Microbiol, Vancouver, BC V6T 1Z3, Canada
[3] Univ British Columbia, Dept Biochem, Vancouver, BC V6T 1Z3, Canada
来源
CHEMISTRY & BIOLOGY | 2004年 / 11卷 / 10期
基金
加拿大自然科学与工程研究理事会;
关键词
D O I
10.1016/j.chembiol.2004.08.011
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The causative agent of severe acute respiratory syndrome (SARS) has been identified as a novel coronavirus, SARS-CoV. The main proteinase of SARS-CoV, 3CL(pro), is an attractive target for therapeutics against SARS owing to its fundamental role in viral replication. We sought to identify novel inhibitors of 3CL(pro) to advance the development of appropriate therapies in the treatment of SARS. 3CL(pro) was cloned, expressed, and purified from the Tor2 isolate. A quenched fluorescence resonance energy transfer assay was developed for 3CL(pro) to screen the proteinase against 50,000 drug-like small molecules on a fully automated system. The primary screen identified 572 hits; through a series of virtual and experimental filters, this number was reduced to five novel small molecules that show potent inhibitory activity (IC50 = 0.5-7 muM) toward SARS-CoV 3CL(pro).
引用
收藏
页码:1445 / 1453
页数:9
相关论文
共 52 条
[41]  
WEBB JL, 1966, ENZYME METABOLIC INH, V3
[42]  
*WHO, 2004, CHIN CONF SARS INF P
[43]  
*WHO, 2004, NEW CAS LAB CONF SAR
[44]  
*WHO, 2003, AL VER PUBL HLTH MAN
[45]  
World Health Organization (WHO), 2003, SUMM TABL SARS CAS C
[46]   Small molecules targeting severe acute respiratory syndrome human coronavirus [J].
Wu, CY ;
Jan, JT ;
Ma, SH ;
Kuo, CJ ;
Juan, HF ;
Cheng, YSE ;
Hsu, HH ;
Huang, HC ;
Wu, D ;
Brik, A ;
Liang, FS ;
Liu, RS ;
Fang, JM ;
Chen, ST ;
Liang, PH ;
Wong, CH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (27) :10012-10017
[47]  
Xiong B, 2003, ACTA PHARMACOL SIN, V24, P497
[48]   HIV protease inhibitor nelfinavir inhibits replication of SARS-associated coronavirus [J].
Yamamoto, N ;
Yang, RG ;
Yoshinaka, Y ;
Amari, S ;
Nakano, T ;
Cinatl, J ;
Rabenau, H ;
Doerr, HW ;
Hunsmann, G ;
Otaka, A ;
Tamamura, H ;
Fujii, N ;
Yamamoto, N .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2004, 318 (03) :719-725
[49]   The crystal structures of severe acute respiratory syndrome virus main protease and its complex with an inhibitor [J].
Yang, HT ;
Yang, MJ ;
Ding, Y ;
Liu, YW ;
Lou, ZY ;
Zhou, Z ;
Sun, L ;
Mo, LJ ;
Ye, S ;
Pang, H ;
Gao, GF ;
Anand, K ;
Bartlam, M ;
Hilgenfeld, R ;
Rao, ZH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (23) :13190-13195
[50]   A simple statistical parameter for use in evaluation and validation of high throughput screening assays [J].
Zhang, JH ;
Chung, TDY ;
Oldenburg, KR .
JOURNAL OF BIOMOLECULAR SCREENING, 1999, 4 (02) :67-73