Antiviral Compounds Screening Targeting HBx Protein of the Hepatitis B Virus

Abstract

A functional cure of hepatitis B virus (HBV) infection or HB antigen loss is rarely achieved by nucleos(t)ide analogs which target viral polymerase. HBx protein is a regulatory protein associated with HBV replication. We thought to identify antiviral compounds targeting HBx protein by analyzing HBx binding activity. Recombinant GST-tagged HBx protein was applied on an FDA-approved drug library chip including 1018 compounds to determine binding affinity by surface plasmon resonance imaging (SPRi) using a PlexArray HT system. GST protein alone was used for control experiments. Candidate compounds were tested for anti-HBV activity as well as cell viability using HepG2.2.15.7 cells and HBV-infected human hepatocytes. Of the 1018 compounds screened, 24 compounds showed binding to HBx protein. Of the top 6 compounds with high affinity to HBx protein, tranilast was found to inhibit HBV replication without affecting cell viability using HepG2.2.15.7 cells. Tranilast also inhibited HBV infection using cultured human hepatocytes. Tranilast reduced HB antigen level dose-dependently. Overall, theSPRi screening assay identified novel drug candidates targeting HBx protein. Tranilast and its related compounds warrant further investigation for the treatment of HBV infection.

Molecular Mechanism of Chronic Viral and Non-Viral Liver Diseases

“…Oncogenic potential of HBV is associated with the X gene product (HBx), which may be able to activate a cellular transcription factor at both viral and cellular promotor sequences in hepatocytes [ 6 ]. HBx may be a regulatory protein associated with HBV replication [ 7 ]. Ma et al screened 1018 FDA-approved compounds and measured HBx-binding ability using a surface plasmon resonance imaging (SPRi) screening assay.…”