1. Chlorcyclizine Inhibits Viral Fusion of Hepatitis C Virus Entry by Directly Targeting HCV Envelope Glycoprotein 1.
- Author
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Hu Z, Rolt A, Hu X, Ma CD, Le DJ, Park SB, Houghton M, Southall N, Anderson DE, Talley DC, Lloyd JR, Marugan JC, and Liang TJ
- Subjects
- Antiviral Agents chemical synthesis, Antiviral Agents chemistry, Antiviral Agents pharmacology, Binding Sites, Biotin chemistry, Diazomethane chemistry, Drug Resistance, Viral drug effects, Genotype, Hepacivirus drug effects, Hepacivirus genetics, Humans, Membrane Fusion drug effects, Molecular Docking Simulation, Piperazines metabolism, Piperazines pharmacology, Ultraviolet Rays, Viral Envelope Proteins metabolism, Virus Internalization drug effects, Hepacivirus metabolism, Piperazines chemistry, Viral Envelope Proteins antagonists & inhibitors
- Abstract
Chlorcyclizine (CCZ) is a potent hepatitis C virus (HCV) entry inhibitor, but its molecular mechanism is unknown. Here, we show that CCZ directly targets the fusion peptide of HCV E1 and interferes with the fusion process. Generation of CCZ resistance-associated substitutions of HCV in vitro revealed six missense mutations in the HCV E1 protein, five being in the putative fusion peptide. A viral fusion assay demonstrated that CCZ blocked HCV entry at the membrane fusion step and that the mutant viruses acquired resistance to CCZ's action in blocking membrane fusion. UV cross-linking of photoactivatable CCZ-diazirine-biotin in both HCV-infected cells and recombinant HCV E1/E2 protein demonstrated direct binding to HCV E1 glycoprotein. Mass spectrometry analysis revealed that CCZ cross-linked to an E1 sequence adjacent to the putative fusion peptide. Docking simulations demonstrate a putative binding model, wherein CCZ binds to a hydrophobic pocket of HCV E1 and forms extensive interactions with the fusion peptide., Competing Interests: Declaration of Interests The authors declare no competing interests., (Published by Elsevier Ltd.)
- Published
- 2020
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