Stable benzotriazole esters as mechanism-based inactivators of the severe acute respiratory syndrome 3CL protease.
Severe acute respiratory syndrome (SARS) is caused by a newly emerged coronavirus that infected more than 8000 individuals and resulted in more than 800 fatalities in 2003. Currently, there is no effective treatment for this epidemic. SARS-3CL(pro) has been shown to be essential for replication and is thus a target for drug discovery. Here, a class of stable benzotriazole esters was reported as mechanism-based inactivators of 3CL(pro), and the most potent inactivator exhibited a k(inact) of 0.0011 s(-1) and a K(i) of 7.5 nM. Mechanistic investigation with kinetic and mass spectrometry analyses indicates that the active site Cys145 is acylated, and that no irreversible inactivation was observed with the use of the C145A mutant. In addition, a noncovalent, competitive inhibition became apparent by using benzotriazole ester surrogates in which the bridged ester-oxygen group is replaced with carbon.
Wu
Chung-Yi
CY
The Genomics Research Center and Institute of Biological Chemistry, Academia Sinica No. 128, Academia Road Section 2, Nan-Kang, Taipei, 115, Taiwan.
King
Ke-Yung
KY
Kuo
Chih-Jung
CJ
Fang
Jim-Min
JM
Wu
Ying-Ta
YT
Ho
Ming-Yi
MY
Liao
Chung-Lin
CL
Shie
Jiun-Jie
JJ
Liang
Po-Huang
PH
Wong
Chi-Huey
CH
D001665:Binding Sites; D000086782:Coronavirus 3C Proteases; D003545:Cysteine / Q000737:chemistry; D003546:Cysteine Endopeptidases; D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D004952:Esters / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008826:Microbial Sensitivity Tests; D008956:Models, Chemical; D011480:Protease Inhibitors / Q000494:pharmacology / Q000627:therapeutic use; D045169:Severe Acute Respiratory Syndrome / Q000188:drug therapy; D013379:Substrate Specificity; D014230:Triazoles / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use; D014764:Viral Proteins / Q000037:antagonists & inhibitors*
D004952:Esters; D011480:Protease Inhibitors; D014230:Triazoles; D014764:Viral Proteins; C012771:benzotriazole; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D000086782:Coronavirus 3C Proteases; D003545:Cysteine