Identification of a small compound that specifically inhibits Zika virus in vitro and in vivo by targeting the NS2B-NS3 protease.
Identification of a small compound that specifically inhibits Zika virus in vitro and in vivo by targeting the NS2B-NS3 protease.
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DOI：
10.1016/j.antiviral.2022.105255
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发表时间：
2022-02
影响因子：
7.6
通讯作者：
Juan Miao;Honggen Yuan;Jingwei Rao;Jiahui Zou;Kelu Yang;Guiqing Peng;S. Cao;Huanchun Chen;Yunfeng Song
中科院分区：
文献类型：
--
作者：
Juan Miao;Honggen Yuan;Jingwei Rao;Jiahui Zou;Kelu Yang;Guiqing Peng;S. Cao;Huanchun Chen;Yunfeng Song
寨卡病毒（ZIKV）已迅速成为全球性威胁，但目前没有ZIKV特异性疫苗或药物可用。在本研究中，从含有4，452个化合物片段的文库中筛选ZIKV NS 2B-NS 3蛋白酶的抑制剂。其中一种化合物6-溴-1，2-萘二酮对ZIKV NS 2B-NS 3蛋白酶表现出高特异性抑制，但对其他病毒蛋白酶没有抑制作用。微量热泳（MST）测定证实化合物以12.26 μM的结合常数（Kd）结合ZIKV NS 2B-NS 3蛋白。间接免疫荧光试验、蛋白质印迹和空斑试验表明，该化合物抑制细胞中的病毒复制。当化合物以1 μM存在时，病毒滴度降低超过75%。时间的添加试验表明，抑制发生在病毒复制阶段，但不是在吸附或入侵阶段。该化合物对HeLa、Vero和BHK-21细胞的半数细胞毒性浓度（CC 50）分别为445.44 μM、123.87 μM和123.64 μM。用该化合物处理的小鼠显示脑、睾丸和卵巢中的组织病理学病变减少。病毒RNA、IL-1β和IL-6 mRNA水平在这些组织中显著降低。总之，本研究已经鉴定了对ZIKV具有高度和特异性抑制作用的小化合物。该化合物可用作治疗剂，也是药物优化的理想起点。
Zika virus (ZIKV) has rapid become a global threat, but no ZIKV-specific vaccines or drugs are currently available. In this study, inhibitors of ZIKV NS2B-NS3 protease were screened from a library containing 4,452 compound fragments. One of the compounds, 6-bromo-1,2-naphthalenedione, exhibited high specific inhibition against ZIKV NS2B-NS3 protease, but had no inhibitory effects against other viral proteases. A microscale thermophoresis (MST) assay confirmed that the compound bound to ZIKV NS2B-NS3 protein with a binding constant (Kd) of 12.26 μM. Indirect immunofluorescence assays, Western blots, and plaque assays indicated that the compound inhibited virus replication in cells. Virus titer was reduced by more than 75% when the compound was present at 1 μM. A time-of-addition assay showed that inhibition occurred at the virus replication stage, but not at the adsorption or invasion stages. The half cytotoxicity concentration (CC50) of the compound on HeLa, Vero, and BHK-21 cells were 445.44 μM, 123.87 μM, and 123.64 μM, respectively.In vivotests using infected AG129 mice demonstrated that treatment with the compound reduced mortality by up to 60%. Mice treated with the compound showed a reduction in histopathological lesions in brain, testis, and ovary. Viral RNA, IL-1β, and IL-6 mRNA levels decreased significantly in these tissues. In summary, this study has identified a small compound with high and specific inhibitory effects on ZIKV. The compound can be used as a therapeutic agent and is also an ideal starting point for drug optimization.