Cao Wenyue, Cho Chia-Chuan Dean, Geng Zhi Zachary, Shaabani Namir, Ma Xinyu R, Vatansever Erol C, Alugubelli Yugendar R, Ma Yuying, Chaki Sankar P, Ellenburg William H, Yang Kai S, Qiao Yuchen, Allen Robert, Neuman Benjamin W, Ji Henry, Xu Shiqing, Liu Wenshe Ray
Texas A&M Drug Discovery Laboratory, Department of Chemistry, Texas A&M University, College Station, Texas 77843, United States.
Sorrento Therapeutics, Inc., San Diego, California 92121, United States.
ACS Cent Sci. 2022 Feb 23;8(2):192-204. doi: 10.1021/acscentsci.1c00910. Epub 2022 Feb 2.
As an essential enzyme of SARS-CoV-2, main protease (M) triggers acute toxicity to its human cell host, an effect that can be alleviated by an M inhibitor. Using this toxicity alleviation, we developed an effective method that allows a bulk analysis of the cellular potency of M inhibitors. This novel assay is advantageous over an antiviral assay in providing precise cellular M inhibition information to assess an M inhibitor. We used this assay to analyze 30 known M 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 M inhibition potency implicating their roles in interfering with key steps other than just the M 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 M inhibition IC value of 31 nM that matches closely to its strong antiviral effect with an EC value of 30 nM. Therefore, we cautiously suggest exploring MPI8 further for COVID-19 preclinical tests.
作为严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的一种关键酶，主要蛋白酶（M）会对其人类细胞宿主引发急性毒性，而一种M抑制剂可以减轻这种效应。利用这种毒性减轻作用，我们开发了一种有效的方法，可对M抑制剂的细胞活性进行批量分析。这种新型检测方法在提供精确的细胞M抑制信息以评估M抑制剂方面优于抗病毒检测方法。我们使用该检测方法分析了30种已知的M抑制剂。与它们高达10 μM的强效抗病毒作用相反，11a、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无法检测到的细胞M抑制活性，这暗示了它们在干扰严重急性呼吸综合征冠状病毒2生命周期中除M催化之外的关键步骤中所起的作用。我们的结果还表明，MPI5、MPI6、MPI7和MPI8具有高细胞活性和抗病毒活性。作为所有测试化合物中细胞活性和抗病毒活性最高的一种，MPI8具有显著的细胞M抑制IC值31 nM，与其30 nM的强效抗病毒作用紧密匹配。因此，我们谨慎建议进一步探索MPI8用于2019冠状病毒病（COVID-19）的临床前测试。
