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. 1996 Feb;40(2):448–453. doi: 10.1128/aac.40.2.448

2',3'-dideoxy-beta-L-5-fluorocytidine inhibits duck hepatitis B virus reverse transcription and suppresses viral DNA synthesis in hepatocytes, both in vitro and in vivo.

F Zoulim 1, E Dannaoui 1, C Borel 1, O Hantz 1, T S Lin 1, S H Liu 1, C Trépo 1, Y C Cheng 1
PMCID: PMC163132  PMID: 8834896

Abstract

beta-L-Nucleoside analogs represent a new class of potent antiviral agents with low cytotoxicity which provide new hope in the therapy of chronic hepatitis B virus (HBV) infections. We evaluated the anti-HBV activity of 2',3'-dideoxy-beta-L-5-fluorocytidine (beta-L-F-ddC), a beta-L-nucleoside analog derived from 2',3'-dideoxycytidine (ddC), in the duck HBV (DHBV) model. This compound was previously shown to inhibit HBV DNA synthesis in a stably transfected hepatoma cell line (F2215). Using a cell-free system for the expression of an enzymatically active DHBV polymerase, we could demonstrate that the triphosphate form of beta-L-F-ddC does inhibit hepadnavirus reverse transcription. In primary duck hepatocyte culture, beta-L-F-ddC showed a potent inhibitory effect on DHBV DNA synthesis which was concentration dependent. Although beta-L-F-ddC was shown to be less active than ddC against the DHBV reverse transcriptase in vitro, beta-L-F-ddC was a stronger inhibitor in hepatocytes. The oral administration of beta-L-F-ddC in experimentally infected ducklings showed that beta-L-F-ddC is a potent inhibitor of viral replication in vivo. Short-term therapy could not prevent a rebound of viral replication after the drug was withdrawn. Preventive therapy with beta-L-F-ddC could delay the onset of viremia by only 1 day compared with the time to the onset of viremia in the control group. The in vivo inhibitory effect of beta-L-F-ddC was much stronger than that of ddC and was not associated with signs of toxicity. Our data show that beta-L-F-ddC inhibits hepadnavirus reverse transcription and is a strong inhibitor of viral replication both in vitro and in vivo.

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Selected References

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  1. Chang C. N., Doong S. L., Zhou J. H., Beach J. W., Jeong L. S., Chu C. K., Tsai C. H., Cheng Y. C., Liotta D., Schinazi R. Deoxycytidine deaminase-resistant stereoisomer is the active form of (+/-)-2',3'-dideoxy-3'-thiacytidine in the inhibition of hepatitis B virus replication. J Biol Chem. 1992 Jul 15;267(20):13938–13942. [PubMed] [Google Scholar]
  2. Chen C. H., Cheng Y. C. Delayed cytotoxicity and selective loss of mitochondrial DNA in cells treated with the anti-human immunodeficiency virus compound 2',3'-dideoxycytidine. J Biol Chem. 1989 Jul 15;264(20):11934–11937. [PubMed] [Google Scholar]
  3. Condreay L. D., Jansen R. W., Powdrill T. F., Johnson L. C., Selleseth D. W., Paff M. T., Daluge S. M., Painter G. R., Furman P. A., Ellis M. N. Evaluation of the potent anti-hepatitis B virus agent (-) cis-5-fluoro-1-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine in a novel in vivo model. Antimicrob Agents Chemother. 1994 Mar;38(3):616–619. doi: 10.1128/aac.38.3.616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Doong S. L., Tsai C. H., Schinazi R. F., Liotta D. C., Cheng Y. C. Inhibition of the replication of hepatitis B virus in vitro by 2',3'-dideoxy-3'-thiacytidine and related analogues. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8495–8499. doi: 10.1073/pnas.88.19.8495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fourel I., Cullen J. M., Saputelli J., Aldrich C. E., Schaffer P., Averett D. R., Pugh J., Mason W. S. Evidence that hepatocyte turnover is required for rapid clearance of duck hepatitis B virus during antiviral therapy of chronically infected ducks. J Virol. 1994 Dec;68(12):8321–8330. doi: 10.1128/jvi.68.12.8321-8330.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fourel I., Gripon P., Hantz O., Cova L., Lambert V., Jacquet C., Watanabe K., Fox J., Guillouzo C., Trepo C. Prolonged duck hepatitis B virus replication in duck hepatocytes cocultivated with rat epithelial cells: a useful system for antiviral testing. Hepatology. 1989 Aug;10(2):186–191. doi: 10.1002/hep.1840100211. [DOI] [PubMed] [Google Scholar]
  7. Fourel I., Li J., Hantz O., Jacquet C., Fox J. J., Trépo C. Effects of 2'-fluorinated arabinosyl-pyrimidine nucleosides on duck hepatitis B virus DNA level in serum and liver of chronically infected ducks. J Med Virol. 1992 Jun;37(2):122–126. doi: 10.1002/jmv.1890370209. [DOI] [PubMed] [Google Scholar]
  8. Fourel I., Saputelli J., Schaffer P., Mason W. S. The carbocyclic analog of 2'-deoxyguanosine induces a prolonged inhibition of duck hepatitis B virus DNA synthesis in primary hepatocyte cultures and in the liver. J Virol. 1994 Feb;68(2):1059–1065. doi: 10.1128/jvi.68.2.1059-1065.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Goff S., Traktman P., Baltimore D. Isolation and properties of Moloney murine leukemia virus mutants: use of a rapid assay for release of virion reverse transcriptase. J Virol. 1981 Apr;38(1):239–248. doi: 10.1128/jvi.38.1.239-248.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kassianides C., Hoofnagle J. H., Miller R. H., Doo E., Ford H., Broder S., Mitsuya H. Inhibition of duck hepatitis B virus replication by 2',3'-dideoxycytidine. A potent inhibitor of reverse transcriptase. Gastroenterology. 1989 Nov;97(5):1275–1280. doi: 10.1016/0016-5085(89)91699-5. [DOI] [PubMed] [Google Scholar]
  11. Lambert V., Chassot S., Kay A., Trepo C., Cova L. In vivo neutralization of duck hepatitis B virus by antibodies specific to the N-terminal portion of pre-S protein. Virology. 1991 Nov;185(1):446–450. doi: 10.1016/0042-6822(91)90796-e. [DOI] [PubMed] [Google Scholar]
  12. Lee B., Luo W. X., Suzuki S., Robins M. J., Tyrrell D. L. In vitro and in vivo comparison of the abilities of purine and pyrimidine 2',3'-dideoxynucleosides to inhibit duck hepadnavirus. Antimicrob Agents Chemother. 1989 Mar;33(3):336–339. doi: 10.1128/aac.33.3.336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lin T. S., Luo M. Z., Liu M. C., Pai S. B., Dutschman G. E., Cheng Y. C. Antiviral activity of 2',3'-dideoxy-beta-L-5-fluorocytidine (beta-L-FddC) and 2',3'-dideoxy-beta-L-cytidine (beta-L-ddC) against hepatitis B virus and human immunodeficiency virus type 1 in vitro. Biochem Pharmacol. 1994 Jan 20;47(2):171–174. doi: 10.1016/0006-2952(94)90002-7. [DOI] [PubMed] [Google Scholar]
  14. Lin T. S., Luo M. Z., Liu M. C., Pai S. B., Dutschman G. E., Cheng Y. C. Synthesis and biological evaluation of 2',3'-dideoxy-L-pyrimidine nucleosides as potential antiviral agents against human immunodeficiency virus (HIV) and hepatitis B virus (HBV). J Med Chem. 1994 Mar 18;37(6):798–803. doi: 10.1021/jm00032a013. [DOI] [PubMed] [Google Scholar]
  15. Marcellin P., Ouzan D., Degos F., Brechot C., Metman E. H., Degott C., Chevalier M., Berthelot P., Trepo C., Benhamou J. P. Randomized controlled trial of adenine arabinoside 5'-monophosphate in chronic active hepatitis B: comparison of the efficacy in heterosexual and homosexual patients. Hepatology. 1989 Sep;10(3):328–331. doi: 10.1002/hep.1840100313. [DOI] [PubMed] [Google Scholar]
  16. Mason W. S., Cullen J., Saputelli J., Wu T. T., Liu C., London W. T., Lustbader E., Schaffer P., O'Connell A. P., Fourel I. Characterization of the antiviral effects of 2' carbodeoxyguanosine in ducks chronically infected with duck hepatitis B virus. Hepatology. 1994 Feb;19(2):398–411. [PubMed] [Google Scholar]
  17. Mason W. S., Taylor J. M. Experimental systems for the study of hepadnavirus and hepatitis delta virus infections. Hepatology. 1989 Apr;9(4):635–645. doi: 10.1002/hep.1840090420. [DOI] [PubMed] [Google Scholar]
  18. Perrillo R. P., Schiff E. R., Davis G. L., Bodenheimer H. C., Jr, Lindsay K., Payne J., Dienstag J. L., O'Brien C., Tamburro C., Jacobson I. M. A randomized, controlled trial of interferon alfa-2b alone and after prednisone withdrawal for the treatment of chronic hepatitis B. The Hepatitis Interventional Therapy Group. N Engl J Med. 1990 Aug 2;323(5):295–301. doi: 10.1056/NEJM199008023230503. [DOI] [PubMed] [Google Scholar]
  19. Popper H., Shafritz D. A., Hoofnagle J. H. Relation of the hepatitis B virus carrier state to hepatocellular carcinoma. Hepatology. 1987 Jul-Aug;7(4):764–772. doi: 10.1002/hep.1840070425. [DOI] [PubMed] [Google Scholar]
  20. Summers J., Smith P. M., Horwich A. L. Hepadnavirus envelope proteins regulate covalently closed circular DNA amplification. J Virol. 1990 Jun;64(6):2819–2824. doi: 10.1128/jvi.64.6.2819-2824.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tuttleman J. S., Pugh J. C., Summers J. W. In vitro experimental infection of primary duck hepatocyte cultures with duck hepatitis B virus. J Virol. 1986 Apr;58(1):17–25. doi: 10.1128/jvi.58.1.17-25.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ueda K., Tsurimoto T., Nagahata T., Chisaka O., Matsubara K. An in vitro system for screening anti-hepatitis B virus drugs. Virology. 1989 Mar;169(1):213–216. doi: 10.1016/0042-6822(89)90057-3. [DOI] [PubMed] [Google Scholar]
  23. Van Draanen N. A., Tisdale M., Parry N. R., Jansen R., Dornsife R. E., Tuttle J. V., Averett D. R., Koszalka G. W. Influence of stereochemistry on antiviral activities and resistance profiles of dideoxycytidine nucleosides. Antimicrob Agents Chemother. 1994 Apr;38(4):868–871. doi: 10.1128/aac.38.4.868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wang G. H., Seeger C. The reverse transcriptase of hepatitis B virus acts as a protein primer for viral DNA synthesis. Cell. 1992 Nov 13;71(4):663–670. doi: 10.1016/0092-8674(92)90599-8. [DOI] [PubMed] [Google Scholar]
  25. Wang G. H., Zoulim F., Leber E. H., Kitson J., Seeger C. Role of RNA in enzymatic activity of the reverse transcriptase of hepatitis B viruses. J Virol. 1994 Dec;68(12):8437–8442. doi: 10.1128/jvi.68.12.8437-8442.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zoulim F., Seeger C. Reverse transcription in hepatitis B viruses is primed by a tyrosine residue of the polymerase. J Virol. 1994 Jan;68(1):6–13. doi: 10.1128/jvi.68.1.6-13.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Zoulim F., Trépo C. Nucleoside analogs in the treatment of chronic viral hepatitis. Efficiency and complications. J Hepatol. 1994 Aug;21(2):142–144. doi: 10.1016/s0168-8278(05)80386-1. [DOI] [PubMed] [Google Scholar]

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