Lee Richard Kuan-Lin, Li Tian-Neng, Chang Sui-Yuan, Chao Tai-Ling, Kuo Chun-Hsien, Pan Max Yu-Chen, Chiou Yu-Ting, Liao Kuan-Ju, Yang Yi, Wu Yi-Hsuan, Huang Chen-Hao, Juan Hsueh-Fen, Hsieh Hsing-Pang, Wang Lily Hui-Ching
Institute of Molecular and Cellular Biology, College of Life Science, National Tsing Hua University, Hsinchu 300013, Taiwan.
SMOBIO Technology, Inc., Hsinchu 300096, Taiwan.
Int J Mol Sci. 2022 Apr 6;23(7):4050. doi: 10.3390/ijms23074050.
Entry inhibitors against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are urgently needed to control the outbreak of coronavirus disease 2019 (COVID-19). This study developed a robust and straightforward assay that detected the molecular interaction between the receptor-binding domain (RBD) of viral spike protein and the angiotensin-converting enzyme 2 (ACE2) receptor in just 10 min. A drug library of 1068 approved compounds was used to screen for SARS-CoV2 entry inhibition, and 9 active drugs were identified as specific pseudovirus entry inhibitors. A plaque reduction neutralization test using authentic SARS-CoV-2 virus in Vero E6 cells confirmed that 2 of these drugs (Etravirine and Dolutegravir) significantly inhibited the infection of SARS-CoV-2. With molecular docking, we showed that both Etravirine and Dolutegravir are preferentially bound to primary ACE2-interacting residues on the RBD domain, implying that these two drug blocks may prohibit the viral attachment of SARS-CoV-2. We compared the neutralizing activities of these entry inhibitors against different pseudoviruses carrying spike proteins from alpha, beta, gamma, and delta variants. Both Etravirine and Dolutegravir showed similar neutralizing activities against different variants, with EC50 values between 4.5 to 5.8 nM for Etravirine and 10.2 to 22.9 nM for Dolutegravir. These data implied that Etravirine and Dolutegravir may serve as general spike inhibitors against dominant viral variants of SARS-CoV-2.
迫切需要针对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的进入抑制剂来控制2019冠状病毒病（COVID-19）的爆发。本研究开发了一种强大且简便的检测方法，可在短短10分钟内检测病毒刺突蛋白的受体结合域（RBD）与血管紧张素转换酶2（ACE2）受体之间的分子相互作用。使用包含1068种获批化合物的药物库筛选SARS-CoV-2进入抑制作用，鉴定出9种活性药物为特异性假病毒进入抑制剂。在Vero E6细胞中使用真实的SARS-CoV-2病毒进行蚀斑减少中和试验证实，其中2种药物（依曲韦林和多替拉韦）可显著抑制SARS-CoV-2的感染。通过分子对接，我们表明依曲韦林和多替拉韦均优先结合于RBD结构域上与ACE2相互作用的主要残基，这意味着这两种药物阻断剂可能会阻止SARS-CoV-2的病毒附着。我们比较了这些进入抑制剂对携带来自α、β、γ和δ变体刺突蛋白的不同假病毒的中和活性。依曲韦林和多替拉韦对不同变体均表现出相似的中和活性，依曲韦林的EC50值在4.5至5.8 nM之间，多替拉韦的EC50值在10.2至22.9 nM之间。这些数据表明依曲韦林和多替拉韦可能作为针对SARS-CoV-2主要病毒变体的通用刺突抑制剂。
