A Novel Capsid Binding Inhibitor Displays Potent Antiviral Activity against Enterovirus D68
A Novel Capsid Binding Inhibitor Displays Potent Antiviral Activity against Enterovirus D68
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DOI：
10.1021/acsinfecdis.9b00284
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发表时间：
2019-11-01
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：
Ma, Chunlong;Hu, Yanmei;Wang, Jun
肠病毒D68 （EV-D68）是一种呼吸道病毒病原体，主要感染8岁以下儿童。虽然EV-D68感染通常会导致中度至重度呼吸道疾病，但近年来，EV-D68引发的神经系统并发症（如急性弛缓性脊髓炎（AFM））的病例越来越多。目前没有针对EV-D68的疫苗或抗病毒药物；因此，我们的目标是开发针对EV-D68的有效和特异性小分子抗病毒药物。在这项研究中，我们报告了我们发现的一种病毒衣壳抑制剂R856932，可以抑制多种当代EVD68菌株，具有个位数到亚微摩尔的功效。机制研究表明，四氮化合物R856932与病毒衣壳蛋白VP1的疏水袋结合，从而阻止病毒在感染细胞中脱衣和释放病毒基因组。R856932的作用机制通过添加时间、Western blot、RT-qPCR、病毒热灭活、病毒连续传代和反向遗传学实验得到证实。位于VP1的单个突变A129V赋予了对R856932的抗性。然而，竞争生长试验显示，与野生型EV-D68病毒相比，编码VP1-A129V的重组病毒的复制适应度似乎有所降低。总的来说，本研究中发现的hit化合物R856932代表了一个有希望的起点，其作用机制已得到确认，可以进一步开发成EV-D68抗病毒药物。
Enterovirus D68 (EV-D68) is a respiratory viral pathogen that primarily infects children under the age of 8. Although EV-D68 infection typically leads to moderate to severe respiratory illnesses, recent years have seen increasing cases of EV-D68 triggered neurological complications such as acute flaccid myelitis (AFM). There is currently no vaccine or antiviral available for EV-D68; we therefore aimed to develop potent and specific small molecule antivirals against EV-D68. In this study, we report our discovery of a viral capsid inhibitor R856932 that inhibits multiple contemporary EVD68 strains with single-digit to submicromolar efficacy. Mechanistic studies have shown that the tetrazole compound R856932 binds to the hydrophobic pocket of viral capsid protein VP1, thereby preventing viral uncoating and release of viral genome in the infected cells. The mechanism of action of R856932 was confirmed by time-of-addition, Western blot, RT-qPCR, viral heat inactivation, serial viral passage, and reverse genetics experiments. A single mutation located at VP1, A129V, confers resistance against R856932. However, a recombination virus encoding VP1-A129V appeared to have compromised fitness of replication compared to the wild-type EV-D68 virus as shown by the competition growth assay. Overall, the hit compound identified in this study, R856932, represents a promising starting point with a confirmed mechanism of action that can be further developed into EV-D68 antivirals.