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| Screening and Inhibition Kinetics of SARS Coronavirus Main Protease Inhibitors |
| ZHU Yun-peng1,2, WANG Peng2, XIA Bo-ran1, TANG Yan-ting2, WANG Quan2 |
1. College of Biological Engineering, Tianjin University of Science and Technology, Tianjin 300457, China;
2. Tianjin International Joint Academy of Biomedicine, Tianjin 300457, China |
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Abstract Heterologous recombinant expression and purification of SARS coronavirus main protease (SARS-CoV Mpro) , Then utilized it as the target, used drug screening model in vitro based on fluorescence resonance energy transfer (FRET), evaluated inhibition activity of 96 compounds from the protease inhibitor focused library, and explored the inhibition of compounds to SARS-CoV Mpro from the perspective of kinetics. The result show that 5 compounds are obtained of which inhibition rate >80%, quenching rate <20% by screened, they are P-1-08,P-1-19,P-2-24,P-2-28 and P-2-54. Their half effective inhibitory concentration (IC50) are 0.69±0.05μmol/L、1.19±0.41μmol/L、0.14±0.01μmol/L、1.36±0.07μmol/L、0.36±0.03μmol/L, respectively. The inhibition of P-1-08, P-1-19, P-2-24 and P-2-54 to SARS-CoV Mpro is irreversible. P-2-28 is reversible inhibition, according to Dixon diagram and Lineweaver-Burk curve found it is competitive inhibition for SARS-CoV Mpro, the inhibition constant Ki is 0.81μmol/L. Through the study about relationship of substrate concentration, IC50 and Ki, validated P-2-28 was competitive inhibition. The discovery of this inhibitor set up the foundation of resource to SARS-CoV Mpro inhibitors, and provided a lead compound for the development of anti-SARS virus drug.
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Received: 21 October 2015
Published: 09 December 2015
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