Identification of a potent and specific retinoic acid-inducible gene 1 pathway activator as a Hepatitis B Virus antiviral through a novel cell-based reporter assay

• Published in: Journal of virological methods Vol. 325; p. 114875
• Main Authors: Shi, Liping, Guo, Guangyang, Zhou, Jinying, Cheng, Zhanling, Zhu, Ren, Kukolj, George, Li, Chris
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-04-2024
• Subjects: Antiviral activity; Cell-based reporter assay; Hepatitis B virus; RIG-I pathway; Small molecule antivirals; Hepatitis B virus; RIG-I pathway; Cell-based reporter assay; Antiviral activity; Small molecule antivirals
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2023.114875

Chronic Hepatitis B Virus (HBV) infection remains a global burden. To identify small molecule RIG-I agonists as antivirals against HBV, we developed an HBV-pgRNA-based interferon-β (IFN-β) luciferase reporter assay with high level of assay sensitivity, specificity and robustness. Through HTS screening, lead compound (JJ#1) was identified to activate RIG-I signaling pathway by inducing TBK1 phosphorylation. Knockdown experiments demonstrated that JJ#1-induced retinoic acid-inducible gene 1 (RIG-I) signaling pathway activation was MAVS-dependent. Furthermore, JJ#1 exhibited HBV antiviral potency in HBV-infected cell models by reducing HBV DNA and antigens (HBsAg and HBeAg).
•An HBV-pgRNA-based IFN-β luciferase reporter assay was established to screen for small molecule activators of the RIG-I pathway with anti-HBV activity.•HTS screening with a small molecule diversity library was performed and lead compound JJ#1 was identified to activate the RIG-I signaling pathway.•Knockdown experiments demonstrated that JJ#1 triggered the RIG-I signaling pathway in a MAVS-dependent manner.•JJ#1 exhibited antiviral activity in HBV-infected cellular models.