Hand, foot, and mouth disease (HFMD) is a common pediatric illness mainly caused by enteroviruses, which are important human pathogens. Currently, there are no available antiviral agents for the therapy of enterovirus infection. In this study, an excellent high-content antiviral screening system utilizing the EV-A71-eGFP reporter virus was developed. Using this screening system, we screened a drug library containing 1042 natural compounds to identify potential EV-A71 inhibitors. Fangchinoline (FAN), a bis-benzylisoquinoline alkaloid, exhibits potential inhibitory effects against various enteroviruses that cause HFMD, such as EV-A71, CV-A10, CV-B3 and CV-A16. Further investigations revealed that FAN targets the early stage of the enterovirus life cycle. Through the selection of FAN-resistant EV-A71 viruses, we demonstrated that the VP1 protein could be a potential target of FAN, as two mutations in VP1 (E145G and V258I) resulted in viral resistance to FAN. Our research suggests that FAN is an efficient inhibitor of EV-A71 and has the potential to be a broad-spectrum antiviral drug against human enteroviruses.

中文翻译：

---

基于报告病毒的高内涵筛选鉴定防己诺林为广谱肠道病毒抑制剂


手足口病（HFMD）是一种常见的儿科疾病，主要由肠道病毒引起，肠道病毒是人类重要的病原体。目前，尚无可用的抗病毒药物用于治疗肠道病毒感染。在这项研究中，开发了一种利用 EV-A71-eGFP 报告病毒的优秀高内涵抗病毒筛选系统。使用该筛选系统，我们筛选了包含 1042 种天然化合物的药物库，以鉴定潜在的 EV-A71 抑制剂。 Fangchinoline (FAN) 是一种双苄基异喹啉生物碱，对引起手足口病的多种肠道病毒（如 EV-A71、CV-A10、CV-B3 和 CV-A16）具有潜在的抑制作用。进一步的研究表明，FAN 针对肠道病毒生命周期的早期阶段。通过选择抗 FAN 的 EV-A71 病毒，我们证明 VP1 蛋白可能是 FAN 的潜在靶标，因为 VP1 的两个突变（E145G 和 V258I）导致病毒对 FAN 产生抗性。我们的研究表明，FAN 是 EV-A71 的有效抑制剂，并有潜力成为针对人类肠道病毒的广谱抗病毒药物。