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. 1995 Aug;39(8):1802–1808. doi: 10.1128/aac.39.8.1802

Mode of action of (R)-9-[4-hydroxy-2-(hydroxymethyl)butyl]guanine against herpesviruses.

D M Lowe 1, W K Alderton 1, M R Ellis 1, V Parmar 1, W H Miller 1, G B Roberts 1, J A Fyfe 1, R Gaillard 1, P Ertl 1, W Snowden 1, et al.
PMCID: PMC162829  PMID: 7486922

Abstract

The activity, metabolism, and mode of action of (R)-9-[4-hydroxy-2-(hydroxymethyl)butyl]guanine (H2G) against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) and varicella-zoster virus (VZV) were studied. Compared to acyclovir (ACV), H2G has superior activity against VZV (50% inhibitory concentration of 2.3 microM) and Epstein-Barr virus (50% inhibitory concentration of 0.9 microM), comparable activity against HSV-1, and weaker activity against HSV-2. The antiviral effect on HSV-1 showed persistence after removal of compound. H2G was metabolized to its mono-, di- and triphosphate derivatives in virus-infected cells, with H2G-triphosphate being the predominant product. Only small amounts of H2G-triphosphate were detected in uninfected cells (1 to 10 pmol/10(6) cells), whereas the level in HSV-1-infected cells reached 1,900 pmol/10(6) cells. H2G was a substrate for all three viral thymidine kinases and could also be phosphorylated by mitochondrial deoxyguanosine kinase. The intracellular half-life of H2G-triphosphate varied in uninfected (2.5 h) and infected (HSV-1, 14 h; VZV, 3.7 h) cells but was always longer than the half-life of ACV-triphosphate (1 to 2 h). H2G-triphosphate inhibited HSV-1, HSV-2, and VZV DNA polymerases competitively with dGTP (Ki of 2.8, 2.2, and 0.3 microM, respectively) but could not replace dGTP as a substrate in a polymerase assay. H2G was not an obligate chain terminator but would only support limited DNA chain extension. Only very small amounts of radioactivity, which were too low to be identified by high-performance liquid chromatography analysis of the digested DNA, could be detected in purified DNA from uninfected cells incubated with [3H]H2G. Thus, H2G acts as an anti-herpesvirus agent, particularly potent against VZV, by formation of high concentrations of relatively stable H2G-triphosphate, which is a potent inhibitor of the viral DNA polymerases.

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Selected References

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  1. Abele G., Eriksson B., Harmenberg J., Wahren B. Inhibition of varicella-zoster virus-induced DNA polymerase by a new guanosine analog, 9-[4-hydroxy-2-(hydroxymethyl)butyl]guanine triphosphate. Antimicrob Agents Chemother. 1988 Aug;32(8):1137–1142. doi: 10.1128/aac.32.8.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abele G., Karlström A., Harmenberg J., Shigeta S., Larsson A., Lindborg B., Wahren B. Inhibiting effect of (RS)-9-[4-hydroxy-2-(hydroxymethyl)butyl]guanine on varicella-zoster virus replication in cell culture. Antimicrob Agents Chemother. 1987 Jan;31(1):76–80. doi: 10.1128/aac.31.1.76. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Akesson-Johansson A., Harmenberg J., Wahren B., Linde A. Inhibition of human herpesvirus 6 replication by 9-[4-hydroxy-2-(hydroxymethyl)butyl]guanine (2HM-HBG) and other antiviral compounds. Antimicrob Agents Chemother. 1990 Dec;34(12):2417–2419. doi: 10.1128/aac.34.12.2417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Averett D. R., Koszalka G. W., Fyfe J. A., Roberts G. B., Purifoy D. J., Krenitsky T. A. 6-Methoxypurine arabinoside as a selective and potent inhibitor of varicella-zoster virus. Antimicrob Agents Chemother. 1991 May;35(5):851–857. doi: 10.1128/aac.35.5.851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chen M. S., Prusoff W. H. Association of thymidylate kinase activity with pyrimidine deoxyribonucleoside kinase induced by herpes simplex virus. J Biol Chem. 1978 Mar 10;253(5):1325–1327. [PubMed] [Google Scholar]
  6. Cieśla J. M., Stolarski R., Shugar D. Cyclic phosphates of some antiviral acyclonucleosides: relationship between conformation and substrate/inhibitor properties in some enzyme systems. Acta Biochim Pol. 1993;40(2):251–260. [PubMed] [Google Scholar]
  7. Crumpacker C. S., Schnipper L. E., Zaia J. A., Levin M. J. Growth inhibition by acycloguanosine of herpesviruses isolated from human infections. Antimicrob Agents Chemother. 1979 May;15(5):642–645. doi: 10.1128/aac.15.5.642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Denizot F., Lang R. Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods. 1986 May 22;89(2):271–277. doi: 10.1016/0022-1759(86)90368-6. [DOI] [PubMed] [Google Scholar]
  9. Earnshaw D. L., Bacon T. H., Darlison S. J., Edmonds K., Perkins R. M., Vere Hodge R. A. Mode of antiviral action of penciclovir in MRC-5 cells infected with herpes simplex virus type 1 (HSV-1), HSV-2, and varicella-zoster virus. Antimicrob Agents Chemother. 1992 Dec;36(12):2747–2757. doi: 10.1128/aac.36.12.2747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Furman P. A., de Miranda P., St Clair M. H., Elion G. B. Metabolism of acyclovir in virus-infected and uninfected cells. Antimicrob Agents Chemother. 1981 Oct;20(4):518–524. doi: 10.1128/aac.20.4.518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fyfe J. A. Differential phosphorylation of (E)-5-(2-bromovinyl)-2'-deoxyuridine monophosphate by thymidylate kinases from herpes simplex viruses types 1 and 2 and varicella zoster virus. Mol Pharmacol. 1982 Mar;21(2):432–437. [PubMed] [Google Scholar]
  12. Fyfe J. A., McKee S. A., Keller P. M. Altered thymidine-thymidylate kinases from strains of herpes simplex virus with modified drug sensitivities to acyclovir and (E)-5-(2-bromovinyl)-2'-deoxyuridine. Mol Pharmacol. 1983 Sep;24(2):316–323. [PubMed] [Google Scholar]
  13. Ilsley D. D., Lee S. H., Miller W. H., Kuchta R. D. Acyclic guanosine analogs inhibit DNA polymerases alpha, delta, and epsilon with very different potencies and have unique mechanisms of action. Biochemistry. 1995 Feb 28;34(8):2504–2510. doi: 10.1021/bi00008a014. [DOI] [PubMed] [Google Scholar]
  14. Lake-Bakaar D. M., Abele G., Lindborg B., Soike K. F., Datema R. Pharmacokinetics and antiviral activity in simian varicella virus-infected monkeys of (R,S)-9-[4-hydroxy-2-(hydroxymethyl) butyl]guanine, an anti-varicella-zoster virus drug. Antimicrob Agents Chemother. 1988 Dec;32(12):1807–1812. doi: 10.1128/aac.32.12.1807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Larsson A., Stenberg K., Ericson A. C., Haglund U., Yisak W. A., Johansson N. G., Oberg B., Datema R. Mode of action, toxicity, pharmacokinetics, and efficacy of some new antiherpesvirus guanosine analogs related to buciclovir. Antimicrob Agents Chemother. 1986 Oct;30(4):598–605. doi: 10.1128/aac.30.4.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Paff M. T., Averett D. R., Prus K. L., Miller W. H., Nelson D. J. Intracellular metabolism of (-)- and (+)-cis-5-fluoro-1-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine in HepG2 derivative 2.2.15 (subclone P5A) cells. Antimicrob Agents Chemother. 1994 Jun;38(6):1230–1238. doi: 10.1128/aac.38.6.1230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Parker W. B., White E. L., Shaddix S. C., Ross L. J., Buckheit R. W., Jr, Germany J. M., Secrist J. A., 3rd, Vince R., Shannon W. M. Mechanism of inhibition of human immunodeficiency virus type 1 reverse transcriptase and human DNA polymerases alpha, beta, and gamma by the 5'-triphosphates of carbovir, 3'-azido-3'-deoxythymidine, 2',3'-dideoxyguanosine and 3'-deoxythymidine. A novel RNA template for the evaluation of antiretroviral drugs. J Biol Chem. 1991 Jan 25;266(3):1754–1762. [PubMed] [Google Scholar]
  18. Roberts G. B., Fyfe J. A., Gaillard R. K., Short S. A. Mutant varicella-zoster virus thymidine kinase: correlation of clinical resistance and enzyme impairment. J Virol. 1991 Dec;65(12):6407–6413. doi: 10.1128/jvi.65.12.6407-6413.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Roberts G. B., Fyfe J. A., McKee S. A., Rahim S. G., Daluge S. M., Almond M. R., Rideout J. L., Koszalka G. W., Krenitsky T. A. Varicella-zoster virus thymidine kinase. Characterization and substrate specificity. Biochem Pharmacol. 1993 Dec 14;46(12):2209–2218. doi: 10.1016/0006-2952(93)90611-y. [DOI] [PubMed] [Google Scholar]
  20. Soike K. F., Bohm R., Huang J. L., Oberg B. Efficacy of (-)-9-[4-hydroxy-2-(hydroxymethyl)butyl]guanine in African green monkeys infected with simian varicella virus. Antimicrob Agents Chemother. 1993 Jun;37(6):1370–1372. doi: 10.1128/aac.37.6.1370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Spector T., Cleland W. W. Meanings of Ki for conventional and alternate-substrate inhibitors. Biochem Pharmacol. 1981 Jan 1;30(1):1–7. doi: 10.1016/0006-2952(81)90277-x. [DOI] [PubMed] [Google Scholar]
  22. Ståhle L., Oberg B. Pharmacokinetics and distribution over the blood brain barrier of two acyclic guanosine analogs in rats, studied by microdialysis. Antimicrob Agents Chemother. 1992 Feb;36(2):339–342. doi: 10.1128/aac.36.2.339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wang L., Karlsson A., Arnér E. S., Eriksson S. Substrate specificity of mitochondrial 2'-deoxyguanosine kinase. Efficient phosphorylation of 2-chlorodeoxyadenosine. J Biol Chem. 1993 Oct 25;268(30):22847–22852. [PubMed] [Google Scholar]

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