Development of a Fluorescence Polarization Assay for the SARS-CoV-2 Papain-like Protease.

IF 4.9 Q1 CHEMISTRY, MEDICINAL

ACS Pharmacology and Translational Science Pub Date : 2025-02-26 eCollection Date: 2025-03-14 DOI:10.1021/acsptsci.4c00642

Kan Li, Prakash Jadhav, Yu Wen, Haozhou Tan, Jun Wang

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引用次数: 0

Abstract

The COVID-19 pandemic has caused significant losses to the global community. Although effective vaccination and antiviral therapeutics provide primary defense, SARS-CoV-2 remains a public health threat, given the emerging resistant variants. The SARS-CoV-2 papain-like protease (PL^pro) is essential for viral replication and is a promising drug target. We recently designed a series of biarylphenyl PL^pro inhibitors with a representative lead Jun12682 showing potent antiviral efficacy in a SARS-CoV-2 infection mouse model. In this study, we designed a fluorescein-labeled biarylphenyl probe Jun12781 and used it to optimize a fluorescence polarization (FP) assay. The FP assay is suitable for high-throughput screening with a Z' factor of 0.69. In addition, we found a positive correlation between the FP binding affinity and the enzymatic inhibitory potency of PL^pro inhibitors, suggesting that the FP assay is valid in characterizing the binding affinity of PL^pro inhibitors.

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SARS-CoV-2木瓜样蛋白酶荧光偏振检测方法的建立。

新冠肺炎疫情给国际社会造成重大损失。尽管有效的疫苗接种和抗病毒治疗提供了主要防御，但鉴于新出现的耐药变体，SARS-CoV-2仍然是一种公共卫生威胁。SARS-CoV-2木瓜蛋白酶（PLpro）对病毒复制至关重要，是一个有希望的药物靶点。我们最近设计了一系列biarylphenyl PLpro抑制剂，具有代表性的先导物Jun12682在SARS-CoV-2感染小鼠模型中显示出有效的抗病毒功效。在这项研究中，我们设计了一个荧光素标记的联芳基苯基探针Jun12781，并用它来优化荧光偏振（FP）测定。FP法适用于高通量筛选，Z′因子为0.69。此外，我们发现FP结合亲和力与PLpro抑制剂的酶抑制效力呈正相关，表明FP测定在表征PLpro抑制剂的结合亲和力方面是有效的。

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来源期刊

ACS Pharmacology and Translational Science Medicine-Pharmacology (medical)

CiteScore

10.00

自引率

3.30%

发文量

133

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