Identification of fangchinoline as a broad-spectrum enterovirus inhibitor through reporter virus based high-content screening

Virol Sin. 2024 Apr;39(2):301-308. doi: 10.1016/j.virs.2024.02.006. Epub 2024 Mar 5.

Abstract

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.

Keywords: EV-A71; Enterovirus; Fangchinoline (FAN); Hand, foot, and mouth disease (HFMD); High-content screening.

MeSH terms

  • Antiviral Agents* / pharmacology
  • Benzylisoquinolines* / pharmacology
  • Capsid Proteins / antagonists & inhibitors
  • Capsid Proteins / genetics
  • Cell Line
  • Drug Evaluation, Preclinical
  • Drug Resistance, Viral* / genetics
  • Enterovirus / drug effects
  • Enterovirus / genetics
  • Enterovirus A, Human / drug effects
  • Enterovirus A, Human / genetics
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics
  • High-Throughput Screening Assays
  • Humans
  • Virus Replication / drug effects

Substances

  • Antiviral Agents
  • Benzylisoquinolines
  • fangchinoline
  • Capsid Proteins
  • Green Fluorescent Proteins