Ogino Minako, Fedorov Yuriy, Adams Drew J, Okada Kazuma, Ito Naoto, Sugiyama Makoto, Ogino Tomoaki
Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
Small Molecule Drug Development Core, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
Viruses. 2019 Sep 14;11(9):856. doi: 10.3390/v11090856.
Vesicular stomatitis virus (VSV) represents a promising platform for developing oncolytic viruses, as well as vaccines against significant human pathogens. To safely control VSV infection in humans, small-molecule drugs that selectively inhibit VSV infection may be needed. Here, using a cell-based high-throughput screening assay followed by an in vitro transcription assay, compounds with a 7-hydroxy-6-methyl-3,4-dihydroquinolin-2(1H)-one structure and an aromatic group at position 4 (named vesiculopolins, VPIs) were identified as VSV RNA polymerase inhibitors. The most effective compound, VPI A, inhibited VSV-induced cytopathic effects and in vitro mRNA synthesis with micromolar to submicromolar 50% inhibitory concentrations. VPI A was found to inhibit terminal de novo initiation rather than elongation for leader RNA synthesis, but not mRNA capping, with the VSV L protein, suggesting that VPI A is targeted to the polymerase domain in the L protein. VPI A inhibited transcription of Chandipura virus, but not of human parainfluenza virus 3, suggesting that it specifically acts on vesiculoviral L proteins. These results suggest that VPIs may serve not only as molecular probes to elucidate the mechanisms of transcription of vesiculoviruses, but also as lead compounds to develop antiviral drugs against vesiculoviruses and other related rhabdoviruses.
水泡性口炎病毒(VSV)是开发溶瘤病毒以及针对重要人类病原体的疫苗的一个有前景的平台。为了安全地控制人类的VSV感染,可能需要选择性抑制VSV感染的小分子药物。在此,通过基于细胞的高通量筛选试验,随后进行体外转录试验,鉴定出具有7-羟基-6-甲基-3,4-二氢喹啉-2(1H)-酮结构且在4位带有芳基的化合物(命名为水泡多聚蛋白,VPI)为VSV RNA聚合酶抑制剂。最有效的化合物VPI A以微摩尔至亚微摩尔的50%抑制浓度抑制VSV诱导的细胞病变效应和体外mRNA合成。发现VPI A抑制前导RNA合成的末端从头起始而非延伸,但不抑制VSV L蛋白的mRNA加帽,这表明VPI A靶向L蛋白中的聚合酶结构域。VPI A抑制钱德普尔病毒的转录,但不抑制人副流感病毒3的转录,这表明它特异性作用于水泡病毒的L蛋白。这些结果表明,VPI不仅可作为阐明水泡病毒转录机制的分子探针,还可作为开发针对水泡病毒和其他相关弹状病毒的抗病毒药物的先导化合物。