Novel Hepatitis B Virus Capsid-Targeting Antiviral That Aggregates Core Particles and Inhibits Nuclear Entry of Viral Cores

• Type of Material: Article
• Authors: Andrew D. Huber, University of Missouri; Dallas L. Pineda, University of Missouri; Dandan Liu, University of Missouri; Kelsey N. Boschert, University of Missouri; Anna T. Gres, University of Missouri; Jennifer J. Wolf, University of Missouri; Emily M. Coonrod, University of Missouri; Jing Tang, University of Minnesota; Thomas G. Laughlin, University of Missouri; Qiongying Yang, University of Missouri; Maritza N. Puray-Chavez, University of Missouri; Juan Ji, University of Missouri; Kamalendra Singh, University of Missouri; Karen Kirby, Emory University; Zhengqiang Wang, University of Minnesota; Stefan Sarafianos, Emory University
• Language: English
• Date: 2019-05-01
• Publisher: ASC Publications
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2018 American Chemical Society
• Final Published Version (URL): http://dx.doi.org/10.1021/acsinfecdis.8b00235
• Title of Journal or Parent Work: ACS Infectious Disease
• Volume: 5
• Issue: 5
• Start Page: 750
• End Page: 758
• Grant/Funding Information: The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.; This work was supported in part by grants from the National Institutes of Health (NIH) (AI100890 and AI121315 to S.G.S.and Z.W.) and Trail to a Cure.; A.D.H. is supported by NIH AI100890- S1, M.N.P.-C. is supported by the Fulbright Student Program, and J.J.W. is supported by a Life Sciences Fellowship from the University of Missouri.; We also acknowledge partial support from Kumamoto University and the Japan Agency for Medical Research and Development.
• Keywords: Pharmacology & Pharmacy; thermal shift assays; Responses; Therapy; Science & Technology; hepatitis B virus; Guidelines; phenylpropenamide derivatives; antiviral; Life Sciences & Biomedicine; Infectious Diseases; Replication; HBV; Infection; capsid; CCCDNA; Chemistry, Medicinal; Management
• Research Categories: Health Sciences, Immunology; Chemistry, Biochemistry; Biology, Virology; Biology, Molecular; Health Sciences, Epidemiology

Abstract

An estimated 240 million are chronically infected with hepatitis B virus (HBV), which can lead to liver disease, cirrhosis, and hepatocellular carcinoma. Currently, HBV treatment options include only nucleoside reverse transcriptase inhibitors and the immunomodulatory agent interferon alpha, and these treatments are generally not curative. New treatments with novel mechanisms of action, therefore, are highly desired for HBV therapy. The viral core protein (Cp) has gained attention as a possible therapeutic target because of its vital roles in the HBV life cycle. Several classes of capsid assembly effectors (CAEs) have been described in detail, and these compounds all increase capsid assembly rate but inhibit HBV replication by different mechanisms. In this study, we have developed a thermal shift-based screening method for CAE discovery and characterization, filling a much-needed gap in high-throughput screening methods for capsid-targeting molecules. Using this approach followed by cell-based screening, we identified the compound HF9C6 as a CAE with low micromolar potency against HBV replication. HF9C6 caused large multicapsid aggregates when capsids were assembled in vitro and analyzed by transmission electron microscopy. Interestingly, when HBV-expressing cells were treated with HF9C6, Cp was excluded from cell nuclei, suggesting that this compound may inhibit nuclear entry of Cp and capsids. Furthermore, mutational scanning of Cp suggested that HF9C6 binds the known CAE binding pocket, indicating that key Cp-compound interactions within this pocket have a role in determining the CAE mechanism of action.