Boceprevir, Calpain Inhibitors II and XII, and GC-376 Have Broad-Spectrum Antiviral Activity against Coronaviruses
Boceprevir, Calpain Inhibitors II and XII, and GC-376 Have Broad-Spectrum Antiviral Activity against Coronaviruses
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DOI：
10.1021/acsinfecdis.0c00761
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发表时间：
2021-03-01
影响因子：
5.3
通讯作者：
Wang, Jun
中科院分区：
文献类型：
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作者：
Hu, Yanmei;Ma, Chunlong;Wang, Jun
随着COVID-19疫情持续发展，发病率及死亡率与日俱增。目前急需对SARS- CoV-2进行有效的治疗。我们最近发现了四种SARS-CoV-2主要蛋白酶（M-pro）抑制剂，包括boceprevir，calpain inhibitors II和XII，以及GC-376，它们在细胞培养中对感染性SARS-CoV-2具有有效的抗病毒活性。在本研究中，我们使用SARS-CoV- 2假病毒中和试验进一步表征了这四种化合物的作用机制。发现GC-376和钙蛋白酶抑制剂II和XII通过抑制Vero细胞中的病毒M-pro和宿主组织蛋白酶L具有双重作用机制。为了排除细胞类型依赖性效应，还使用病毒产量减少测定在表达2型跨膜丝氨酸蛋白酶的Caco-2细胞中证实了这四种化合物对SARS-CoV-2的抗病毒活性。此外，我们发现这四个化合物不仅对SARS-CoV-2具有广谱抗病毒活性，而且对SARS-CoV、MERS-CoV以及人冠状病毒（CoV）229 E、OC 43和NL 63也具有广谱抗病毒活性。热位移结合试验和酶促荧光共振能量转移试验证实了其作用机制是通过靶向病毒M-pro。我们进一步表明，这四种化合物与瑞德西韦联合使用时具有相加的抗病毒作用。总之，这些结果表明，boceprevir，钙蛋白酶抑制剂II和XII以及GC-376可能是进一步开发对抗现有人类冠状病毒以及未来新出现的CoV的有希望的起点。
As the COVID-19 pandemic continues to unfold, the morbidity and mortality are increasing daily. Effective treatment for SARS- CoV-2 is urgently needed. We recently discovered four SARS-CoV-2 main protease (M-pro) inhibitors including boceprevir, calpain inhibitors II and XII, and GC-376 with potent antiviral activity against infectious SARS-CoV-2 in cell culture. In this study, we further characterized the mechanism of action of these four compounds using the SARS- CoV- 2 pseudovirus neutralization assay. It was found that GC-376 and calpain inhibitors II and XII have a dual mechanism of action by inhibiting both viral M-pro and host cathepsin L in Vero cells. To rule out the cell-type dependent effect, the antiviral activity of these four compounds against SARS-CoV-2 was also confirmed in type 2 transmembrane serine protease-expressing Caco-2 cells using the viral yield reduction assay. In addition, we found that these four compounds have broad-spectrum antiviral activity in inhibiting not only SARS-CoV-2 but also SARS-CoV, and MERS-CoV, as well as human coronaviruses (CoVs) 229E, OC43, and NL63. The mechanism of action is through targeting the viral M-pro, which was supported by the thermal shift-binding assay and enzymatic fluorescence resonance energy transfer assay. We further showed that these four compounds have additive antiviral effect when combined with remdesivir. Altogether, these results suggest that boceprevir, calpain inhibitors II and XII, and GC-376 might be promising starting points for further development against existing human coronaviruses as well as future emerging CoVs.