Anti-hepatitis B virus activity in vitro of combinations of tenofovir with nucleoside/nucleotide analogues.
BACKGROUND
Long-term management of some chronic hepatitis B patients might require combination therapy using drugs with distinct resistance profiles to sustain viral suppression and to reduce the resistance-associated failure. Tenofovir disoproxil fumarate (TDF), approved for hepatitis B virus (HBV) and HIV-1 treatment, is active against wildtype HBV and HBV containing YMDD mutations, which confer resistance to emtricitabine (FTC), lamivudine (3TC) and telbivudine (LdT) and contribute to entecavir (ETV) resistance. We therefore evaluated the in vitro anti-HBV activity of tenofovir (TFV), the active parent drug of TDF, combined with FTC, 3TC, ETV, LdT and adefovir (AFV).


METHODS
The anti-HBV activities of the compounds were tested using the AD38 cell line that expresses wild-type HBV from a tetracycline-controllable promoter. Intracellular HBV DNA levels were quantified using real-time PCR assay and cytotoxicities were assessed with XTT assays. The antiviral data of the drug combinations were evaluated using MacSynergy analyses on the basis of the Bliss independence model as well as isobologram analyses on the basis of the Loewe additivity theory.


RESULTS
All drug combinations tested, FTC+TFV, 3TC+TFV, ETV+TFV, LdT+TFV and AFV+TFV, showed additive antiviral interactions as analysed by MacSynergy. Isobologram analyses revealed that these combination pairs were additive, with the exception of FTC+TFV, which demonstrated slight synergistic activity. No cytotoxic or antagonistic effects were observed with any of the combinations tested.


CONCLUSIONS
The combination of TFV with FTC, 3TC, ETV, LdT or AFV had additive to slightly synergistic anti-HBV effects in vitro. These results support the use of TDF as a component in combination regimens with currently available anti-HBV nucleoside analogues.
Gilead Sciences, Inc, Durham, NC, USA. yuao.zhu@gilead.com
Yuao
Y
Zhu
Maria
M
Curtis
Xiaoping
X
Qi
Michael D
MD
Miller
Katyna
K
Borroto-Esoda
D000225:Adenine / Q000008:administration & dosage / Q000031:analogs & derivatives*; D000998:Antiviral Agents / Q000008:administration & dosage*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003841:Deoxycytidine / Q000008:administration & dosage / Q000031:analogs & derivatives; D004305:Dose-Response Relationship, Drug; D004359:Drug Therapy, Combination; D000068679:Emtricitabine; D006147:Guanine / Q000008:administration & dosage / Q000031:analogs & derivatives; D006515:Hepatitis B virus / Q000254:growth & development*; D019694:Hepatitis B, Chronic / Q000188:drug therapy*; D006801:Humans; D019259:Lamivudine / Q000008:administration & dosage; D009705:Nucleosides / Q000008:administration & dosage*; D009711:Nucleotides / Q000008:administration & dosage*; D063065:Organophosphonates / Q000008:administration & dosage*; D011744:Pyrimidinones / Q000008:administration & dosage; D018894:Reverse Transcriptase Inhibitors / Q000008:administration & dosage*; D000077712:Telbivudine; D000068698:Tenofovir; D013936:Thymidine / Q000031:analogs & derivatives
D000998:Antiviral Agents; D009705:Nucleosides; D009711:Nucleotides; D063065:Organophosphonates; D011744:Pyrimidinones; D018894:Reverse Transcriptase Inhibitors; D003841:Deoxycytidine; D000077712:Telbivudine; D019259:Lamivudine; C413685:entecavir; D006147:Guanine; C053001:adefovir; D000068698:Tenofovir; D000068679:Emtricitabine; D000225:Adenine; D013936:Thymidine