Abstract
The (-) and (+) enantiomers of the nucleoside analog cis-5-fluoro-1-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine (2',3'-dideoxy-5-fluoro-3'-thiacytidine; FTC) have been shown to inhibit hepatitis B virus replication in vitro in HepG2 derivative 2.2.15 (subclone P5A) cells. (-)-FTC and (+)-FTC were anabolized to 5'-monophosphate, 5'-diphosphate, and 5'-triphosphate in this cell line. (-)-FTC was more efficiently phosphorylated to the 5'-triphosphate than (+)-FTC, and levels of 3.6 and 0.2 pmol/10(6) cells, respectively, were detected after incubation with 1 microM compound for 24 h. A time course study showed that nucleotides were formed rapidly in a dose-dependent manner and reached a steady-state intracellular concentration by 3 to 6 h. The intracellular half-life of (-)-FTC 5'-triphosphate was 2.4 h. Both (-)- and (+)-FTC were converted to diphosphocholine derivatives, analogous to CDP-choline, but only (+)-FTC was converted to the diphosphoethanolamine derivative, analogous to CDP-ethanolamine. (-)-FTC was not detectably deaminated at either the nucleoside or nucleotide level. (+)-FTC was partially deaminated by these cells. The transport of (-)-and (+)-FTC was examined in HepG2 cells. (+)-FTC enters these cells by way of the nitrobenzylthioinosine-susceptible, equilibrative nucleoside transporter. In contrast, the influx of (-)-FTC was only partially susceptible to inhibitors of nucleoside transport, indicating that (-)-FTC may have multiple transport mechanisms. These metabolic results are consistent with the conclusion that (-)-FTC 5'-triphosphate mediates the anti-hepatitis B virus activity of (-)-FTC.
Full text
PDF








Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ahluwalia G., Cooney D. A., Hartman N. R., Mitsuya H., Yarchoan R., Fridland A., Broder S., Johns D. G. Anomalous accumulation and decay of 2',3'-dideoxyadenosine-5'-triphosphate in human T-cell cultures exposed to the anti-HIV drug 2',3'-dideoxyinosine. Drug Metab Dispos. 1993 Mar-Apr;21(2):369–376. [PubMed] [Google Scholar]
- Ahluwalia G., Cooney D. A., Mitsuya H., Fridland A., Flora K. P., Hao Z., Dalal M., Broder S., Johns D. G. Initial studies on the cellular pharmacology of 2',3'-dideoxyinosine, an inhibitor of HIV infectivity. Biochem Pharmacol. 1987 Nov 15;36(22):3797–3800. doi: 10.1016/0006-2952(87)90440-0. [DOI] [PubMed] [Google Scholar]
- Beasley R. P., Hwang L. Y., Lin C. C., Chien C. S. Hepatocellular carcinoma and hepatitis B virus. A prospective study of 22 707 men in Taiwan. Lancet. 1981 Nov 21;2(8256):1129–1133. doi: 10.1016/s0140-6736(81)90585-7. [DOI] [PubMed] [Google Scholar]
- Blakley R. L., Harwood F. C., Huff K. D. Cytostatic effects of 2',3'-dideoxyribonucleosides on transformed human hemopoietic cell lines. Mol Pharmacol. 1990 Feb;37(2):328–332. [PubMed] [Google Scholar]
- CLELAND W. W. Computer programmes for processing enzyme kinetic data. Nature. 1963 May 4;198:463–465. doi: 10.1038/198463a0. [DOI] [PubMed] [Google Scholar]
- Cammack N., Rouse P., Marr C. L., Reid P. J., Boehme R. E., Coates J. A., Penn C. R., Cameron J. M. Cellular metabolism of (-) enantiomeric 2'-deoxy-3'-thiacytidine. Biochem Pharmacol. 1992 May 28;43(10):2059–2064. doi: 10.1016/0006-2952(92)90162-c. [DOI] [PubMed] [Google Scholar]
- 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]
- Coates J. A., Cammack N., Jenkinson H. J., Mutton I. M., Pearson B. A., Storer R., Cameron J. M., Penn C. R. The separated enantiomers of 2'-deoxy-3'-thiacytidine (BCH 189) both inhibit human immunodeficiency virus replication in vitro. Antimicrob Agents Chemother. 1992 Jan;36(1):202–205. doi: 10.1128/aac.36.1.202. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cooney D. A., Dalal M., Mitsuya H., McMahon J. B., Nadkarni M., Balzarini J., Broder S., Johns D. G. Initial studies on the cellular pharmacology of 2',3-dideoxycytidine, an inhibitor of HTLV-III infectivity. Biochem Pharmacol. 1986 Jul 1;35(13):2065–2068. doi: 10.1016/0006-2952(86)90571-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Furman P. A., Davis M., Liotta D. C., Paff M., Frick L. W., Nelson D. J., Dornsife R. E., Wurster J. A., Wilson L. J., Fyfe J. A. The anti-hepatitis B virus activities, cytotoxicities, and anabolic profiles of the (-) and (+) enantiomers of cis-5-fluoro-1-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine. Antimicrob Agents Chemother. 1992 Dec;36(12):2686–2692. doi: 10.1128/aac.36.12.2686. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gati W. P., Dagnino L., Paterson A. R. Enantiomeric selectivity of adenosine transport systems in mouse erythrocytes and L1210 cells. Biochem J. 1989 Nov 1;263(3):957–960. doi: 10.1042/bj2630957. [DOI] [PMC free article] [PubMed] [Google Scholar]
- HARTMANN K. U., HEIDELBERGER C. Studies on fluorinated pyrimidines. XIII. Inhibition of thymidylate synthetase. J Biol Chem. 1961 Nov;236:3006–3013. [PubMed] [Google Scholar]
- Hao Z., Cooney D. A., Hartman N. R., Perno C. F., Fridland A., DeVico A. L., Sarngadharan M. G., Broder S., Johns D. G. Factors determining the activity of 2',3'-dideoxynucleosides in suppressing human immunodeficiency virus in vitro. Mol Pharmacol. 1988 Oct;34(4):431–435. [PubMed] [Google Scholar]
- Hao Z., Stowe E. E., Ahluwalia G., Baker D. C., Hebbler A. K., Chisena C., Musser S. M., Kelley J. A., Perno C. F., Johns D. G. Characterization of 2',3'-dideoxycytidine diphosphocholine and 2',3'-dideoxycytidine diphosphoethanolamine. Prominent phosphodiester metabolites of the anti-HIV nucleoside 2',3'-dideoxycytidine. Drug Metab Dispos. 1993 Jul-Aug;21(4):738–744. [PubMed] [Google Scholar]
- Ho H. T., Hitchcock M. J. Cellular pharmacology of 2',3'-dideoxy-2',3'-didehydrothymidine, a nucleoside analog active against human immunodeficiency virus. Antimicrob Agents Chemother. 1989 Jun;33(6):844–849. doi: 10.1128/aac.33.6.844. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jansen R. W., Johnson L. C., Averett D. R. High-capacity in vitro assessment of anti-hepatitis B virus compound selectivity by a virion-specific polymerase chain reaction assay. Antimicrob Agents Chemother. 1993 Mar;37(3):441–447. doi: 10.1128/aac.37.3.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kierdaszuk B., Bohman C., Ullman B., Eriksson S. Substrate specificity of human deoxycytidine kinase toward antiviral 2',3'-dideoxynucleoside analogs. Biochem Pharmacol. 1992 Jan 22;43(2):197–206. doi: 10.1016/0006-2952(92)90278-q. [DOI] [PubMed] [Google Scholar]
- Korba B. E., Milman G. A cell culture assay for compounds which inhibit hepatitis B virus replication. Antiviral Res. 1991 Mar-Apr;15(3):217–228. doi: 10.1016/0166-3542(91)90068-3. [DOI] [PubMed] [Google Scholar]
- 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]
- Müller R., Baumgarten R., Markus R., Schulz M., Wittenberg H., Hintsche-Kilger B., Fengler J. D., Von Wussow P., Meisel H., Klein H. Treatment of chronic hepatitis B with interferon alfa-2b. J Hepatol. 1990;11 (Suppl 1):S137–S140. doi: 10.1016/0168-8278(90)90181-p. [DOI] [PubMed] [Google Scholar]
- 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]
- Schinazi R. F., McMillan A., Cannon D., Mathis R., Lloyd R. M., Peck A., Sommadossi J. P., St Clair M., Wilson J., Furman P. A. Selective inhibition of human immunodeficiency viruses by racemates and enantiomers of cis-5-fluoro-1-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine. Antimicrob Agents Chemother. 1992 Nov;36(11):2423–2431. doi: 10.1128/aac.36.11.2423. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sells M. A., Chen M. L., Acs G. Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc Natl Acad Sci U S A. 1987 Feb;84(4):1005–1009. doi: 10.1073/pnas.84.4.1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shewach D. S., Liotta D. C., Schinazi R. F. Affinity of the antiviral enantiomers of oxathiolane cytosine nucleosides for human 2'-deoxycytidine kinase. Biochem Pharmacol. 1993 Apr 6;45(7):1540–1543. doi: 10.1016/0006-2952(93)90058-5. [DOI] [PubMed] [Google Scholar]
- Staprans S., Loeb D. D., Ganem D. Mutations affecting hepadnavirus plus-strand DNA synthesis dissociate primer cleavage from translocation and reveal the origin of linear viral DNA. J Virol. 1991 Mar;65(3):1255–1262. doi: 10.1128/jvi.65.3.1255-1262.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Starnes M. C., Cheng Y. C. Cellular metabolism of 2',3'-dideoxycytidine, a compound active against human immunodeficiency virus in vitro. J Biol Chem. 1987 Jan 25;262(3):988–991. [PubMed] [Google Scholar]
- Stenberg K., Larsson A., Datema R. Metabolism and mode of action of (R)-9-(3,4-dihydroxybutyl)guanine in herpes simplex virus-infected vero cells. J Biol Chem. 1986 Feb 15;261(5):2134–2139. [PubMed] [Google Scholar]
- Törnevik Y., Eriksson S. 2',3'-Dideoxycytidine toxicity in cultured human CEM T lymphoblasts: effects of combination with 3'-azido-3'-deoxythymidine and thymidine. Mol Pharmacol. 1990 Aug;38(2):237–243. [PubMed] [Google Scholar]
- Vijayalakshmi D., Belt J. A. Sodium-dependent nucleoside transport in mouse intestinal epithelial cells. Two transport systems with differing substrate specificities. J Biol Chem. 1988 Dec 25;263(36):19419–19423. [PubMed] [Google Scholar]
- WILKINSON G. N. Statistical estimations in enzyme kinetics. Biochem J. 1961 Aug;80:324–332. doi: 10.1042/bj0800324. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilson J. E., Martin J. L., Borroto-Esoda K., Hopkins S., Painter G., Liotta D. C., Furman P. A. The 5'-triphosphates of the (-) and (+) enantiomers of cis-5-fluoro-1-[2-(hydroxymethyl)-1,3-oxathiolane-5-yl]cytosine equally inhibit human immunodeficiency virus type 1 reverse transcriptase. Antimicrob Agents Chemother. 1993 Aug;37(8):1720–1722. doi: 10.1128/aac.37.8.1720. [DOI] [PMC free article] [PubMed] [Google Scholar]