Evaluation of Single and Combination Therapies with Tenofovir Disoproxil Fumarate and Emtricitabine In Vitro and in a Robust Mouse Model Supporting High Levels of Hepatitis B Virus Replication

• Type of Material: Article
• Authors: Raymond F Schinazi, Emory University; Leda C. Bassit, Emory University; Marcia M. Clayton, Temple University; Bill Sun, Temple University; James J Kohler, Emory University; Aleksandr Obikhod, Emory University; Alla Arzumanyan, Temple University; Mark A. Feitelson, Temple University
• Language: English
• Date: 2012-12
• Publisher: American Society for Microbiology
• Publication Version: Final Published Version
• Copyright Statement: © 2012, American Society for Microbiology. All Rights Reserved.
• Final Published Version (URL): http://aac.asm.org/content/56/12/6186
• Title of Journal or Parent Work: Antimicrobial Agents and Chemotherapy
• Volume: 56
• Issue: 12
• Start Page: 6186
• End Page: 6191
• Grant/Funding Information: This work was primarily supported by NIH grant R01-AI-076535 (to R.F.S. and M.A.F.). This work was partially supported by grants 5R37-AI-41980, 5R37-AI-25899, and 5P30-AI-50409 and the U.S. Department of Veterans Affairs (to R.F.S.), and by NIH grants CA48656, CA66971, and CA104025 (to M.A.F.).
• Research Categories: Biology, Microbiology; Health Sciences, Medicine and Surgery

Abstract

Next-generation therapies for chronic hepatitis B virus (HBV) infection will involve combinations of established and/or experimental drugs. The current study investigated the in vitro and in vivo efficacy of tenofovir disoproxil fumarate (TDF) and/or emtricitabine [(−)-FTC] alone and in combination therapy for HBV infection utilizing the HepAD38 system (human hepatoblastoma cells transfected with HBV). Cellular pharmacology studies demonstrated increased levels of (−)-FTC triphosphate with coincubation of increasing concentrations of TDF, while (−)-FTC had no effect on intracellular tenofovir (TFV) diphosphate levels. Quantification of extracellular HBV by real-time PCR from hepatocytes demonstrated the anti-HBV activity with TDF, (−)-FTC, and their combination. Combination of (−)-FTC with TDF or TFV (ratio, 1:1) had a weighted average combination index of 0.7 for both combination sets, indicating synergistic antiviral effects. No cytotoxic effects were observed with any regimens. Using an in vivo murine model which develops robust HBV viremia in nude mice subcutaneously injected with HepAD38 cells, TDF (33 to 300 mg/kg of body weight/day) suppressed virus replication for up to 10 days posttreatment. At 300 mg/kg/day, (−)-FTC strongly suppressed virus titers to up to 14 days posttreatment. Combination therapy (33 mg/kg/day each drug) sustained suppression of virus titer/ml serum (<1 log10 unit from pretreatment levels) at 14 days posttreatment, while single-drug treatments yielded virus titers 1.5 to 2 log units above the initial virus titers. There was no difference in mean alanine aminotransferase values or mean wet tumor weights for any of the groups, suggesting a lack of drug toxicity. TDF–(−)-FTC combination therapy provides more effective HBV suppression than therapy with each drug alone.