Discovery of SARS-CoV-2 Papain-like Protease Inhibitors through a Combination of High-Throughput Screening and a FlipGFP-Based Reporter Assay.
Discovery of SARS-CoV-2 Papain-like Protease Inhibitors through a Combination of High-Throughput Screening and a FlipGFP-Based Reporter Assay.
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DOI：
10.1021/acscentsci.1c00519
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发表时间：
2021-07-28
影响因子：
18.2
通讯作者：
Wang J
中科院分区：
文献类型：
--
作者：
Ma C;Sacco MD;Xia Z;Lambrinidis G;Townsend JA;Hu Y;Meng X;Szeto T;Ba M;Zhang X;Gongora M;Zhang F;Marty MT;Xiang Y;Kolocouris A;Chen Y;Wang J
SARS-CoV-2的木瓜蛋白酶样蛋白酶（PLpro）是一个有效的抗病毒药物靶点。通过基于荧光共振能量转移的高通量筛选和随后的先导物优化，我们鉴定了几种PLpro抑制剂，包括Jun9 - 72 - 2和Jun9 - 75 - 4，与报道的SARS-CoV PLpro抑制剂GRL 0617相比，其具有改善的酶抑制和抗病毒活性。值得注意的是，我们开发了一种基于细胞的FlipGFP测定法，可用于预测PLpro抑制剂在BSL-2环境中的细胞抗病毒活性。PLpro与GRL 0617复合物的X射线晶体结构显示，GRL 0617与SARS-CoV-2的结合诱导BL 2环的构象变化为更封闭的构象。分子动力学模拟表明，Jun9 - 72 - 2和Jun9 - 75 - 4比GRL 0617参与更广泛的相互作用。总体而言，本研究中鉴定的PLpro抑制剂代表了作为SARS-CoV-2抗病毒药物进一步开发的有希望的候选药物，并且FlipGFP-PLpro测定是在BSL-2环境中筛选PLpro抑制剂的合适替代物。一种基于细胞的FlipGFP报告基因检测被开发用于筛选SARS-CoV-2木瓜蛋白酶样蛋白酶抑制剂，并显示与抗病毒活性呈正相关。
The papain-like protease (PLpro) of SARS-CoV-2 is a validated antiviral drug target. Through a fluorescence resonance energy transfer-based high-throughput screening and subsequent lead optimization, we identified several PLpro inhibitors including Jun9-72-2 and Jun9-75-4 with improved enzymatic inhibition and antiviral activity compared to GRL0617, which was reported as a SARS-CoV PLpro inhibitor. Significantly, we developed a cell-based FlipGFP assay that can be applied to predict the cellular antiviral activity of PLpro inhibitors in the BSL-2 setting. X-ray crystal structure of PLpro in complex with GRL0617 showed that binding of GRL0617 to SARS-CoV-2 induced a conformational change in the BL2 loop to a more closed conformation. Molecular dynamics simulations showed that Jun9-72-2 and Jun9-75-4 engaged in more extensive interactions than GRL0617. Overall, the PLpro inhibitors identified in this study represent promising candidates for further development as SARS-CoV-2 antivirals, and the FlipGFP-PLpro assay is a suitable surrogate for screening PLpro inhibitors in the BSL-2 setting. A cell-based FlipGFP reporter assay was developed for the screening of SARS-CoV-2 papain-like protease inhibitors and was shown to have a positive correlation with antiviral activity.
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DOI：
10.1056/nejmoa2022926
发表时间：
2020-11-19
期刊：
The New England journal of medicine
影响因子：
--
作者：
RECOVERY Collaborative Group;Horby P;Mafham M;Linsell L;Bell JL;Staplin N;Emberson JR;Wiselka M;Ustianowski A;Elmahi E;Prudon B;Whitehouse T;Felton T;Williams J;Faccenda J;Underwood J;Baillie JK;Chappell LC;Faust SN;Jaki T;Jeffery K;Lim WS;Montgomery A;Rowan K;Tarning J;Watson JA;White NJ;Juszczak E;Haynes R;Landray MJ
通讯作者：
Landray MJ
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
SARS-CoV-2 类木瓜蛋白酶的晶体结构
影响因子：
14.5
作者：
Gao, Xiaopan;Qin, Bo;Cui, Sheng
通讯作者：
Cui, Sheng
影响因子：
5.4
作者：
Froggatt HM;Heaton BE;Heaton NS
通讯作者：
Heaton NS
影响因子：
4.4
作者：
FELLER, SE;ZHANG, YH;BROOKS, BR
通讯作者：
BROOKS, BR