Evaluation of SARS-CoV-2 Main Protease Inhibitors Using a Novel Cell-Based Assay.
Evaluation of SARS-CoV-2 Main Protease Inhibitors Using a Novel Cell-Based Assay.
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DOI：
10.1021/acscentsci.1c00910
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发表时间：
2022-02-23
影响因子：
18.2
通讯作者：
Liu WR
中科院分区：
文献类型：
--
作者：
Cao W;Cho CD;Geng ZZ;Shaabani N;Ma XR;Vatansever EC;Alugubelli YR;Ma Y;Chaki SP;Ellenburg WH;Yang KS;Qiao Y;Allen R;Neuman BW;Ji H;Xu S;Liu WR
作为SARS-CoV-2的必需酶，主蛋白酶（MPro）对其人类细胞宿主触发急性毒性，这种作用可以通过MPro抑制剂减轻。使用这种毒性缓解，我们开发了一种有效的方法，允许大量分析MPro抑制剂的细胞效力。这种新的测定在提供精确的细胞MPro抑制信息以评估MPro抑制剂方面优于抗病毒测定。我们使用该测定法分析了30种已知的MPro抑制剂。与其强大的抗病毒作用和高达10 μM相反，11 a、钙蛋白酶抑制剂II、钙蛋白酶XII、依布硒啉、苄普地尔、氯喹和羟氯喹显示出相对较弱至不可检测的细胞MPro抑制效力，暗示它们在SARS-CoV-2生命周期中干扰除MPro催化以外的关键步骤的作用。我们的研究结果还表明，MPI 5，MPI 6，MPI 7和MPI 8具有较高的细胞和抗病毒效力。作为所有测试化合物中具有最高细胞和抗病毒效力的化合物，MPI 8具有31 nM的显著细胞MPro抑制IC 50值，与其30 nM的EC 50值的强抗病毒作用密切匹配。因此，我们谨慎地建议进一步探索MPI 8用于COVID-19临床前试验。建立了一种基于细胞分选的检测方法，对约30种SARS-CoV-2主要蛋白酶抑制剂进行了研究，发现MPI 8是最有效的蛋白酶抑制剂，其他蛋白酶抑制剂可能具有不同的机制。
As an essential enzyme of SARS-CoV-2, main protease (MPro) triggers acute toxicity to its human cell host, an effect that can be alleviated by an MPro inhibitor. Using this toxicity alleviation, we developed an effective method that allows a bulk analysis of the cellular potency of MPro inhibitors. This novel assay is advantageous over an antiviral assay in providing precise cellular MPro inhibition information to assess an MPro inhibitor. We used this assay to analyze 30 known MPro inhibitors. Contrary to their strong antiviral effects and up to 10 μM, 11a, calpain inhibitor II, calpain XII, ebselen, bepridil, chloroquine, and hydroxychloroquine showed relatively weak to undetectable cellular MPro inhibition potency implicating their roles in interfering with key steps other than just the MPro catalysis in the SARS-CoV-2 life cycle. Our results also revealed that MPI5, MPI6, MPI7, and MPI8 have high cellular and antiviral potency. As the one with the highest cellular and antiviral potency among all tested compounds, MPI8 has a remarkable cellular MPro inhibition IC50 value of 31 nM that matches closely to its strong antiviral effect with an EC50 value of 30 nM. Therefore, we cautiously suggest exploring MPI8 further for COVID-19 preclinical tests. A cell sorting-based assay was developed to study about 30 SARS-CoV-2 main protease inhibitors, revealing MPI8 as the most potent one and others with likely different mechanisms.
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