Identification of Compounds Targeting Hepatitis B Virus Core Protein Dimerization through a Split Luciferase Complementation Assay

Antimicrob Agents Chemother. 2018 Nov 26;62(12):e01302-18. doi: 10.1128/AAC.01302-18. Print 2018 Dec.

Abstract

The capsid of the hepatitis B virus is an attractive antiviral target for developing therapies against chronic hepatitis B infection. Currently available core protein allosteric modulators (CpAMs) mainly affect one of the two major types of protein-protein interactions involved in the process of capsid assembly, namely, the interaction between the core dimers. Compounds targeting the interaction between two core monomers have not been rigorously screened due to the lack of screening models. We report here a cell-based assay in which the formation of core dimers is indicated by split luciferase complementation (SLC). Making use of this model, 2 compounds, Arbidol (umifenovir) and 20-deoxyingenol, were identified from a library containing 672 compounds as core dimerization regulators. Arbidol and 20-deoxyingenol inhibit the hepatitis B virus (HBV) DNA replication in vitro by decreasing and increasing the formation of core dimer and capsid, respectively. Our results provided a proof of concept for the cell model to be used to screen new agents targeting the step of core dimer and capsid formation.

Keywords: 20-deoxyingenol; Arbidol; cell model; compound screen; core protein; dimer; hepatitis B virus; split luciferase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antiviral Agents / pharmacology*
  • Capsid / drug effects
  • Capsid / metabolism
  • Capsid / ultrastructure
  • Cell Line
  • DNA Replication / drug effects
  • DNA, Viral / antagonists & inhibitors
  • DNA, Viral / biosynthesis
  • DNA, Viral / genetics
  • Diterpenes / pharmacology*
  • Gene Expression Regulation, Viral*
  • Genes, Reporter
  • HEK293 Cells
  • Hepatitis B virus / drug effects*
  • Hepatitis B virus / genetics
  • Hepatitis B virus / metabolism
  • Hepatocytes / drug effects
  • Hepatocytes / virology
  • High-Throughput Screening Assays
  • Humans
  • Indoles / pharmacology*
  • Luciferases / genetics
  • Luciferases / metabolism
  • Protein Binding / drug effects
  • Protein Multimerization / drug effects*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Viral Core Proteins / antagonists & inhibitors*
  • Viral Core Proteins / genetics
  • Viral Core Proteins / metabolism

Substances

  • Antiviral Agents
  • DNA, Viral
  • Diterpenes
  • Indoles
  • Recombinant Fusion Proteins
  • Viral Core Proteins
  • umifenovir
  • Luciferases