Rosmarinic acid exhibits broad anti-enterovirus A71 activity by inhibiting the interaction between the five-fold axis of capsid VP1 and cognate sulfated receptors

• Published in: Emerging microbes & infections Vol. 9; no. 1; pp. 1194 - 1205
• Main Authors: Hsieh, Chung-Fan, Jheng, Jia-Rong, Lin, Guan-Hua, Chen, Yu-Li, Ho, Jin-Yuan, Liu, Chien-Jou, Hsu, Kuei-Yang, Chen, Yuan-Siao, Chan, Yoke Fun, Yu, Hui-Ming, Hsieh, Pei-Wen, Chern, Jyh-Haur, Horng, Jim-Tong
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis 01-01-2020; Taylor & Francis Ltd; Taylor & Francis Group
• Subjects: Animals; Antiviral Agents - administration & dosage; Antiviral Agents - pharmacology; Capsid Proteins - antagonists & inhibitors; Capsid Proteins - chemistry; Capsid Proteins - genetics; Cell Line; Cinnamates - administration & dosage; Cinnamates - pharmacology; Depsides - administration & dosage; Depsides - pharmacology; Disease Models, Animal; Drug Evaluation, Preclinical; Enterovirus A, Human - drug effects; Enterovirus A, Human - metabolism; Enterovirus A, Human - pathogenicity; Enterovirus A71; Enterovirus Infections - drug therapy; Enterovirus Infections - virology; five-fold axis; Heparan sulfate; Heparitin Sulfate - metabolism; Humans; Jurkat Cells; Membrane Glycoproteins - metabolism; Mice; Mutation; Original; P-selectin glycoprotein ligand-1; Plant Extracts - administration & dosage; Plant Extracts - pharmacology; Protein Binding - drug effects; receptor; Rosmarinic Acid; Salvia miltiorrhiza - chemistry; scavenger receptor B2; Static Electricity; viral entry; Viral infections; Virulence Factors - antagonists & inhibitors; Virulence Factors - chemistry; Virulence Factors - genetics; Viruses; P-selectin glycoprotein ligand-1; heparan sulfate; receptor; scavenger receptor B2; viral entry; rosmarinic acid; five-fold axis; Enterovirus A71
• ISSN: 2222-1751; 2222-1751
• DOI: 10.1080/22221751.2020.1767512

Enterovirus A71 (EV-A71), a positive-stranded RNA virus of the Picornaviridae family, may cause neurological complications or fatality in children. We examined specific factors responsible for this virulence using a chemical genetics approach. Known compounds from an anti-EV-A71 herbal medicine, Salvia miltiorrhiza (Danshen), were screened for anti-EV-A71. We identified a natural product, rosmarinic acid (RA), as a potential inhibitor of EV-A71 by cell-based antiviral assay and in vivo mouse model. Results also show that RA may affect the early stage of viral infection and may target viral particles directly, thereby interfering with virus-P-selectin glycoprotein ligand-1 (PSGL1) and virus-heparan sulfate interactions without abolishing the interaction between the virus and scavenger receptor B2 (SCARB2). Sequencing of the plaque-purified RA-resistant viruses revealed a N104K mutation in the five-fold axis of the structural protein VP1, which contains positively charged amino acids reportedly associated with virus-PSGL1 and virus-heparan sulfate interactions via electrostatic attraction. The plasmid-derived recombinant virus harbouring this mutation was confirmed to be refractory to RA inhibition. Receptor pull-down showed that this non-positively charged VP1-N104 is critical for virus binding to heparan sulfate. As the VP1-N104 residue is conserved among different EV-A71 strains, RA may be useful for inhibiting EV-A71 infection, even for emergent virus variants. Our study provides insight into the molecular mechanism of virus-host interactions and identifies a promising new class of inhibitors based on its antiviral activity and broad spectrum effects against a range of EV-A71.