Inhibition of hepatitis B viral entry by nucleic acid polymers in HepaRG cells and primary human hepatocytes

PLoS One. 2017 Jun 21;12(6):e0179697. doi: 10.1371/journal.pone.0179697. eCollection 2017.


Hepatitis B virus (HBV) infection remains a major public health concern worldwide with 240 million individuals chronically infected and at risk of developing cirrhosis and hepatocellular carcinoma. Current treatments rarely cure chronic hepatitis B infection, highlighting the need for new anti-HBV drugs. Nucleic acid polymers (NAPs) are phosphorothioated oligonucleotides that have demonstrated a great potential to inhibit infection with several viruses. In chronically infected human patients, NAPs administration lead to a decline of blood HBsAg and HBV DNA and to HBsAg seroconversion, the expected signs of functional cure. NAPs have also been shown to prevent infection of duck hepatocytes with the Avihepadnavirus duck hepatitis B virus (DHBV) and to exert an antiviral activity against established DHBV infection in vitro and in vivo. In this study, we investigated the specific anti-HBV antiviral activity of NAPs in the HepaRG human hepatoma cell line and primary cultures of human hepatocytes. NAPs with different chemical features (phosphorothioation, 2'O-methyl ribose, 5-methylcytidine) were assessed for antiviral activity when provided at the time of HBV inoculation or post-inoculation. NAPs dose-dependently inhibited HBV entry in a phosphorothioation-dependent, sequence-independent and size-dependent manner. This inhibition of HBV entry by NAPs was impaired by 2'O-methyl ribose modification. NAP treatment after viral inoculation did not elicit any antiviral activity.

MeSH terms

  • Antiviral Agents / chemistry
  • Antiviral Agents / pharmacology*
  • Antiviral Agents / toxicity
  • Cell Survival / drug effects
  • Cells, Cultured
  • Cytidine / analogs & derivatives
  • Cytidine / chemistry
  • DNA, Viral / blood
  • Hepatitis B Surface Antigens / blood
  • Hepatitis B e Antigens / blood
  • Hepatitis B virus / genetics
  • Hepatitis B virus / physiology*
  • Hepatocytes / cytology
  • Hepatocytes / virology
  • Humans
  • Immunoassay
  • Nucleic Acids / chemistry*
  • Phosphates / chemistry
  • Polymers / chemistry
  • Polymers / pharmacology*
  • Polymers / toxicity
  • Real-Time Polymerase Chain Reaction
  • Virus Internalization / drug effects*


  • Antiviral Agents
  • DNA, Viral
  • Hepatitis B Surface Antigens
  • Hepatitis B e Antigens
  • Nucleic Acids
  • Phosphates
  • Polymers
  • Cytidine
  • phosphorodithioic acid
  • 5-methylcytidine

Grant support

The work was funded by Replicor Inc. The funder provided support in the form of salaries for authors [CG, NM, MB and AV], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.