Skip to main content
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1991 Oct;35(10):2003–2011. doi: 10.1128/aac.35.10.2003

Synergistic inhibition of human immunodeficiency virus type 1 replication in vitro by two-drug and three-drug combinations of 3'-azido-3'-deoxythymidine, phosphonoformate, and 2',3'-dideoxythymidine.

X B Kong 1, Q Y Zhu 1, R M Ruprecht 1, K A Watanabe 1, J M Zeidler 1, J W Gold 1, B Polsky 1, D Armstrong 1, T C Chou 1
PMCID: PMC245315  PMID: 1722077

Abstract

The effects of 3'-azido-3'-deoxythymidine (AZT), phosphonoformate (PFA), and 2',3'-dideoxythymidine (ddT) and their combination on human immunodeficiency type 1 (HIV-1) replication were studied by measuring the HIV-1 p24 antigen expression and reverse transcriptase (RT) release in HIV-1-infected MT4 cells in vitro. RT activity was also measured in a cell-free system by using poly(rA)-oligo(dT) as the primer-template, and cell growth inhibition was measured in noninfected MT4 cells. The interactions of these two- and three-drug combinations were evaluated by the combination index (CI) method and isobologram techniques. The 50% effective concentrations (EC50s) of AZT, PFA, and ddT were 0.014 to 0.005, 9.4 to 8.8, and 8.4 to 2.5 microM, respectively, for p24 enzyme-linked immunosorbent assays (ELISAs) and 0.005 to 0.0034, 1.43 to 1.37, and 2.87 to 2.83 microM, respectively, for RT activity in vitro; for RT activity in the cell-free system, the EC50s were 0.00019 to 0.00024, 0.012 to 0.02, and 0.00074 to 0.0005 microM, for AZT-5'-triphosphate, PFA, and ddT-5'-triphosphate, respectively. AZT in combination with PFA (1:200) or ddT (1:5) as well as the combination of these three drugs (1:200:5) synergistically inhibited HIV-1 replication and RT activity in the cell-free system over a wide range of drug concentrations, with the CIs ranging from 0.5 to 0.09, in which CIs of less than 1, 1, and greater than 1 indicate synergism, additive effect, and antagonism, respectively. Three- and two-drug combinations of AZT, PFA, and ddT showed similar degrees of synergism against HIV-1 replication in p24 assays and RT release assays, whereas the combination of AZT and ddT was found to be the most selective in terms of its anti-HIV-1 effect versus cytotoxicity. Dose reduction indices calculated from both HIV-1 replication inhibition, as measured by p24 ELISA and by RT activity in the cell-free system, indicated that two- and three-drug combinations at high effect levels and the selected combination ratios allow 2- to 240-fold dose reduction over the single drug alone in terms of their anti-HIV-1 effects. The three-drug combination showed the highest dose reduction index. These finding suggest that increased efficacy and reduced toxicity may be achieved in AIDS therapy by using AZT, PFA, and ddT in two- or three-drug combinations.

Full text

PDF
2003

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barré-Sinoussi F., Chermann J. C., Rey F., Nugeyre M. T., Chamaret S., Gruest J., Dauguet C., Axler-Blin C., Vézinet-Brun F., Rouzioux C. Isolation of a T-lymphotropic retrovirus from a patient at risk for acquired immune deficiency syndrome (AIDS). Science. 1983 May 20;220(4599):868–871. doi: 10.1126/science.6189183. [DOI] [PubMed] [Google Scholar]
  2. Broder S., Gallo R. C. A pathogenic retrovirus (HTLV-III) linked to AIDS. N Engl J Med. 1984 Nov 15;311(20):1292–1297. doi: 10.1056/NEJM198411153112006. [DOI] [PubMed] [Google Scholar]
  3. Chou T. C., Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul. 1984;22:27–55. doi: 10.1016/0065-2571(84)90007-4. [DOI] [PubMed] [Google Scholar]
  4. Drach J. C., Thomas M. A., Barnett J. W., Smith S. H., Shipman C., Jr Tritiated thymidine incorporation does not measure DNA synthesis in ribavirin-treated human cells. Science. 1981 May 1;212(4494):549–551. doi: 10.1126/science.7209549. [DOI] [PubMed] [Google Scholar]
  5. Eriksson B. F., Schinazi R. F. Combinations of 3'-azido-3'-deoxythymidine (zidovudine) and phosphonoformate (foscarnet) against human immunodeficiency virus type 1 and cytomegalovirus replication in vitro. Antimicrob Agents Chemother. 1989 May;33(5):663–669. doi: 10.1128/aac.33.5.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Hartshorn K. L., Sandstrom E. G., Neumeyer D., Paradis T. J., Chou T. C., Schooley R. T., Hirsch M. S. Synergistic inhibition of human T-cell lymphotropic virus type III replication in vitro by phosphonoformate and recombinant alpha-A interferon. Antimicrob Agents Chemother. 1986 Jul;30(1):189–191. doi: 10.1128/aac.30.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hartshorn K. L., Vogt M. W., Chou T. C., Blumberg R. S., Byington R., Schooley R. T., Hirsch M. S. Synergistic inhibition of human immunodeficiency virus in vitro by azidothymidine and recombinant alpha A interferon. Antimicrob Agents Chemother. 1987 Feb;31(2):168–172. doi: 10.1128/aac.31.2.168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hayashi S., Fine R. L., Chou T. C., Currens M. J., Broder S., Mitsuya H. In vitro inhibition of the infectivity and replication of human immunodeficiency virus type 1 by combination of antiretroviral 2',3'-dideoxynucleosides and virus-binding inhibitors. Antimicrob Agents Chemother. 1990 Jan;34(1):82–88. doi: 10.1128/aac.34.1.82. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Johnson V. A., Barlow M. A., Chou T. C., Fisher R. A., Walker B. D., Hirsch M. S., Schooley R. T. Synergistic inhibition of human immunodeficiency virus type 1 (HIV-1) replication in vitro by recombinant soluble CD4 and 3'-azido-3'-deoxythymidine. J Infect Dis. 1989 May;159(5):837–844. doi: 10.1093/infdis/159.5.837. [DOI] [PubMed] [Google Scholar]
  11. Johnson V. A., Walker B. D., Barlow M. A., Paradis T. J., Chou T. C., Hirsch M. S. Synergistic inhibition of human immunodeficiency virus type 1 and type 2 replication in vitro by castanospermine and 3'-azido-3'-deoxythymidine. Antimicrob Agents Chemother. 1989 Jan;33(1):53–57. doi: 10.1128/aac.33.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Koshida R., Vrang L., Gilljam G., Harmenberg J., Oberg B., Wahren B. Inhibition of human immunodeficiency virus in vitro by combinations of 3'-azido-3'-deoxythymidine and foscarnet. Antimicrob Agents Chemother. 1989 May;33(5):778–780. doi: 10.1128/aac.33.5.778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Larder B. A., Chesebro B., Richman D. D. Susceptibilities of zidovudine-susceptible and -resistant human immunodeficiency virus isolates to antiviral agents determined by using a quantitative plaque reduction assay. Antimicrob Agents Chemother. 1990 Mar;34(3):436–441. doi: 10.1128/aac.34.3.436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lehoang P., Girard B., Robinet M., Marcel P., Zazoun L., Matheron S., Rozenbaum W., Katlama C., Morer I., Lernestedt J. O. Foscarnet in the treatment of cytomegalovirus retinitis in acquired immune deficiency syndrome. Ophthalmology. 1989 Jun;96(6):865–874. doi: 10.1016/s0161-6420(89)32824-7. [DOI] [PubMed] [Google Scholar]
  15. Mitsuya H., Broder S. Inhibition of the in vitro infectivity and cytopathic effect of human T-lymphotrophic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV) by 2',3'-dideoxynucleosides. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1911–1915. doi: 10.1073/pnas.83.6.1911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mitsuya H., Weinhold K. J., Furman P. A., St Clair M. H., Lehrman S. N., Gallo R. C., Bolognesi D., Barry D. W., Broder S. 3'-Azido-3'-deoxythymidine (BW A509U): an antiviral agent that inhibits the infectivity and cytopathic effect of human T-lymphotropic virus type III/lymphadenopathy-associated virus in vitro. Proc Natl Acad Sci U S A. 1985 Oct;82(20):7096–7100. doi: 10.1073/pnas.82.20.7096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sandstrom E. G., Kaplan J. C., Byington R. E., Hirsch M. S. Inhibition of human T-cell lymphotropic virus type III in vitro by phosphonoformate. Lancet. 1985 Jun 29;1(8444):1480–1482. doi: 10.1016/s0140-6736(85)92255-x. [DOI] [PubMed] [Google Scholar]
  18. Scudiero D. A., Shoemaker R. H., Paull K. D., Monks A., Tierney S., Nofziger T. H., Currens M. J., Seniff D., Boyd M. R. Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines. Cancer Res. 1988 Sep 1;48(17):4827–4833. [PubMed] [Google Scholar]
  19. Spira T. J., Bozeman L. H., Holman R. C., Warfield D. T., Phillips S. K., Feorino P. M. Micromethod for assaying reverse transcriptase of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus. J Clin Microbiol. 1987 Jan;25(1):97–99. doi: 10.1128/jcm.25.1.97-99.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Starnes M. C., Cheng Y. C. Inhibition of human immunodeficiency virus reverse transcriptase by 2',3'-dideoxynucleoside triphosphates: template dependence, and combination with phosphonoformate. Virus Genes. 1989 May;2(3):241–251. doi: 10.1007/BF00125341. [DOI] [PubMed] [Google Scholar]
  21. Vogt M. W., Durno A. G., Chou T. C., Coleman L. A., Paradis T. J., Schooley R. T., Kaplan J. C., Hirsch M. S. Synergistic interaction of 2',3'-dideoxycytidine and recombinant interferon-alpha-A on replication of human immunodeficiency virus type 1. J Infect Dis. 1988 Aug;158(2):378–385. doi: 10.1093/infdis/158.2.378. [DOI] [PubMed] [Google Scholar]
  22. Vogt M. W., Hartshorn K. L., Furman P. A., Chou T. C., Fyfe J. A., Coleman L. A., Crumpacker C., Schooley R. T., Hirsch M. S. Ribavirin antagonizes the effect of azidothymidine on HIV replication. Science. 1987 Mar 13;235(4794):1376–1379. doi: 10.1126/science.2435003. [DOI] [PubMed] [Google Scholar]
  23. Vrang L., Bazin H., Remaud G., Chattopadhyaya J., Oberg B. Inhibition of the reverse transcriptase from HIV by 3'-azido-3'-deoxythymidine triphosphate and its threo analogue. Antiviral Res. 1987 Mar;7(3):139–149. doi: 10.1016/0166-3542(87)90002-7. [DOI] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES