Virus entry is a multistep process. It initiates when the virus attaches to the host cell and ends when the viral contents reach the cytosol. Genetically unrelated viruses can subvert analogous subcellular mechanisms and use similar trafficking pathways for successful entry. Antiviral strategies targeting early steps of infection are therefore appealing, particularly when the probability for successful interference through a common step is highest. We describe here potent inhibitory effects on content release and infection by chimeric vesicular stomatitis virus (VSV) containing the envelope proteins of Zaire ebolavirus (VSV-ZEBOV) or severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (VSV-SARS-CoV-2) elicited by Apilimod and Vacuolin-1, small-molecule inhibitors of the main endosomal phosphatidylinositol-3-phosphate/phosphatidylinositol 5-kinase, PIKfyve. We also describe potent inhibition of SARS-CoV-2 strain 2019-nCoV/USA-WA1/2020 by Apilimod. These results define tools for studying the intracellular trafficking of pathogens elicited by inhibition of PIKfyve kinase and suggest the potential for targeting this kinase in developing small-molecule antivirals against SARS-CoV-2.

病毒进入是一个多步骤的过程。当病毒附着在宿主细胞上时它开始，当病毒内容物到达细胞质时结束。遗传上不相关的病毒可以破坏类似的亚细胞机制，并使用类似的贩运途径成功进入。因此，针对感染早期步骤的抗病毒策略很有吸引力，特别是当通过共同步骤成功干扰的可能性最高时。我们在此描述了含有扎伊尔埃博拉病毒 (VSV-ZEBOV) 包膜蛋白或严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) (VSV-SARS- CoV-2）由 Apilimod 和 Vacuolin-1（主要内体磷脂酰肌醇 3-磷酸/磷脂酰肌醇 5-激酶 PIKfyve 的小分子抑制剂）引发。我们还描述了 Apilimod 对 SARS-CoV-2 毒株 2019-nCoV/USA-WA1/2020 的有效抑制作用。这些结果定义了用于研究通过抑制 PIKfyve 激酶引起的病原体细胞内运输的工具，并表明针对该激酶开发针对 SARS-CoV-2 的小分子抗病毒药物的潜力。