Identification of novel drug scaffolds for inhibition of SARS-CoV 3-Chymotrypsin-like protease using virtual and high-throughput screenings.
Identification of novel drug scaffolds for inhibition of SARS-CoV 3-Chymotrypsin-like protease using virtual and high-throughput screenings.
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使用虚拟和高通量筛选鉴定新的药物支架，以抑制SARS-COV 3-羟丙烯蛋白酶样蛋白酶。
DOI：
10.1016/j.bmc.2013.11.041
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发表时间：
2014-01-01
影响因子：
3.5
通讯作者：
Johnson ME
中科院分区：
文献类型：
--
作者：
Lee H;Mittal A;Patel K;Gatuz JL;Truong L;Torres J;Mulhearn DC;Johnson ME
我们使用虚拟筛选(VS)和高通量筛选(HTS)技术相结合的方法来识别抗人SARS-CoV 3CLPro的新型非肽类小分子抑制剂。一种基于结构的VS方法集成了对接和基于药效团的方法，用于从锌库中计算筛选621,000个化合物。该筛选方案使用已知的3CLPro抑制剂进行了验证，并针对速度、提高的选择性和适应受体的灵活性进行了优化。随后，开发和优化了一种基于荧光的酶促HTS分析方法，从主要基于VS结果选择的四个结构不同的文库中实验筛选约41,000个化合物。通过使用表面等离子体共振(SPR)的二次正交结合分析，消除了初始HTS命中的假阳性。本研究确定了一种可逆的小分子抑制物，其KI值为11.1μM。总之，这些结果验证了我们的方法是筛选虚拟文库和化学文库的合适方法，我们的研究中新发现的化合物为进一步开发SARS-CoV3CLPro抑制剂提供了一个有前途的结构支架。
We have used a combination of virtual screening (VS) and high-throughput screening (HTS) techniques to identify novel, non-peptidic small molecule inhibitors against human SARS-CoV 3CLpro. A structure-based VS approach integrating docking and pharmacophore based methods was employed to computationally screen 621,000 compounds from the ZINC library. The screening protocol was validated using known 3CLpro inhibitors and was optimized for speed, improved selectivity, and for accommodating receptor flexibility. Subsequently, a fluorescence-based enzymatic HTS assay was developed and optimized to experimentally screen approximately 41,000 compounds from four structurally diverse libraries chosen mainly based on the VS results. False positives from initial HTS hits were eliminated by a secondary orthogonal binding analysis using surface plasmon resonance (SPR). The campaign identified a reversible small molecule inhibitor exhibiting mixed-type inhibition with a Ki value of 11.1 μM. Together, these results validate our protocols as suitable approaches to screen virtual and chemical libraries, and the newly identified compound reported in our study represents a promising structural scaffold to pursue for further SARS-CoV 3CLpro inhibitor development.
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影响因子：
21.1
作者：
Chen S;Jonas F;Shen C;Hilgenfeld R
通讯作者：
Hilgenfeld R
影响因子：
5
作者：
Grum-Tokars, Valerie;Ratia, Kiira;Begaye, Adrian;Baker, Susan C.;Mesecar, Andrew D.
通讯作者：
Mesecar, Andrew D.
影响因子：
--
作者：
Ghosh, Arun K.;Xi, Kai;Johnson, Michael E.;Baker, Susan C.;Mesecar, Andrew D.
通讯作者：
Mesecar, Andrew D.
影响因子：
3
作者：
Duan, Y;Wu, C;Kollman, P
通讯作者：
Kollman, P
影响因子：
7.3
作者：
Charifson, PS;Corkery, JJ;Walters, WP
通讯作者：
Walters, WP