Skip to main content
PMC home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1994 Apr;38(4):688–692. doi: 10.1128/aac.38.4.688

Preclinical evaluation of MKC-442, a highly potent and specific inhibitor of human immunodeficiency virus type 1 in vitro.

M Baba 1, S Shigeta 1, S Yuasa 1, H Takashima 1, K Sekiya 1, M Ubasawa 1, H Tanaka 1, T Miyasaka 1, R T Walker 1, E De Clercq 1
PMCID: PMC284526  PMID: 7518216

Abstract

MKC-442 (6-benzyl-1-ethoxymethyl-5-isopropyluracil or I-EBU) has recently been identified as a highly potent and specific inhibitor of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase. Since the compound has favorable pharmacokinetic and toxicity profiles in vivo, we have evaluated MKC-442 for its inhibitory effect on the replication of HIV-1 in various cell cultures, including human peripheral blood lymphocytes and monocyte-macrophages. The 50 and 90% effective concentrations for HIV-1 (HTLV-IIIB strain) replication in MT-4 cells were 15 and 98 nM, respectively. MKC-442 was also inhibitory to HIV-1 replication in peripheral blood lymphocytes and monocyte-macrophages as determined by the production of p24 antigens in the culture supernatant. Fluorescence-activated cell sorter analysis revealed that MKC-442 was equally active against zidovudine-resistant mutants and zidovudine-susceptible strains. Furthermore, combinations of MKC-442 with either 3'-azido-3'-deoxythymidine, 2',3'-dideoxycytidine, or 2',3'-dideoxyinosine synergistically inhibited the replication of HIV-1. Thus, MKC-442 has been considered as a candidate for clinical efficacy studies.

Full text

PDF
688

Selected References

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

  1. Baba M., De Clercq E., Tanaka H., Ubasawa M., Takashima H., Sekiya K., Nitta I., Umezu K., Nakashima H., Mori S. Potent and selective inhibition of human immunodeficiency virus type 1 (HIV-1) by 5-ethyl-6-phenylthiouracil derivatives through their interaction with the HIV-1 reverse transcriptase. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2356–2360. doi: 10.1073/pnas.88.6.2356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baba M., De Clercq E., Tanaka H., Ubasawa M., Takashima H., Sekiya K., Nitta I., Umezu K., Walker R. T., Mori S. Highly potent and selective inhibition of human immunodeficiency virus type 1 by a novel series of 6-substituted acyclouridine derivatives. Mol Pharmacol. 1991 Jun;39(6):805–810. [PubMed] [Google Scholar]
  3. Baba M., Ito M., Shigeta S., Tanaka H., Miyasaka T., Ubasawa M., Umezu K., Walker R. T., De Clercq E. Synergistic inhibition of human immunodeficiency virus type 1 replication by 5-ethyl-1-ethoxymethyl-6-(phenylthio)uracil (E-EPU) and azidothymidine in vitro. Antimicrob Agents Chemother. 1991 Jul;35(7):1430–1433. doi: 10.1128/aac.35.7.1430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baba M., Shigeta S., Tanaka H., Miyasaka T., Ubasawa M., Umezu K., Walker R. T., Pauwels R., De Clercq E. Highly potent and selective inhibition of HIV-1 replication by 6-phenylthiouracil derivatives. Antiviral Res. 1992 Apr;17(4):245–264. doi: 10.1016/0166-3542(92)90021-v. [DOI] [PubMed] [Google Scholar]
  5. Baba M., Tanaka H., De Clercq E., Pauwels R., Balzarini J., Schols D., Nakashima H., Perno C. F., Walker R. T., Miyasaka T. Highly specific inhibition of human immunodeficiency virus type 1 by a novel 6-substituted acyclouridine derivative. Biochem Biophys Res Commun. 1989 Dec 29;165(3):1375–1381. doi: 10.1016/0006-291x(89)92756-3. [DOI] [PubMed] [Google Scholar]
  6. Baba M., Yuasa S., Niwa T., Yamamoto M., Yabuuchi S., Takashima H., Ubasawa M., Tanaka H., Miyasaka T., Walker R. T. Effect of human serum on the in vitro anti-HIV-1 activity of 1-[(2-hydroxyethoxy)methyl]-6-(phenylthio)thymine (HEPT) derivatives as related to their lipophilicity and serum protein binding. Biochem Pharmacol. 1993 Jun 22;45(12):2507–2512. doi: 10.1016/0006-2952(93)90232-l. [DOI] [PubMed] [Google Scholar]
  7. Balzarini J., Karlsson A., Pérez-Pérez M. J., Camarasa M. J., De Clercq E. Knocking-out concentrations of HIV-1-specific inhibitors completely suppress HIV-1 infection and prevent the emergence of drug-resistant virus. Virology. 1993 Oct;196(2):576–585. doi: 10.1006/viro.1993.1513. [DOI] [PubMed] [Google Scholar]
  8. Balzarini J., Karlsson A., Pérez-Pérez M. J., Vrang L., Walbers J., Zhang H., Oberg B., Vandamme A. M., Camarasa M. J., De Clercq E. HIV-1-specific reverse transcriptase inhibitors show differential activity against HIV-1 mutant strains containing different amino acid substitutions in the reverse transcriptase. Virology. 1993 Jan;192(1):246–253. doi: 10.1006/viro.1993.1027. [DOI] [PubMed] [Google Scholar]
  9. Balzarini J., Pérez-Pérez M. J., San-Félix A., Schols D., Perno C. F., Vandamme A. M., Camarasa M. J., De Clercq E. 2',5'-Bis-O-(tert-butyldimethylsilyl)-3'-spiro-5''-(4''-amino-1'',2''- oxathiole-2'',2'-dioxide)pyrimidine (TSAO) nucleoside analogues: highlyselective inhibitors of human immunodeficiency virus type 1 that are targeted at the viral reverse transcriptase. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4392–4396. doi: 10.1073/pnas.89.10.4392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Davey R. T., Jr, Dewar R. L., Reed G. F., Vasudevachari M. B., Polis M. A., Kovacs J. A., Falloon J., Walker R. E., Masur H., Haneiwich S. E. Plasma viremia as a sensitive indicator of the antiretroviral activity of L-697,661. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5608–5612. doi: 10.1073/pnas.90.12.5608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. De Clercq E. HIV-1-specific RT inhibitors: highly selective inhibitors of human immunodeficiency virus type 1 that are specifically targeted at the viral reverse transcriptase. Med Res Rev. 1993 May;13(3):229–258. doi: 10.1002/med.2610130303. [DOI] [PubMed] [Google Scholar]
  13. Debyser Z., Pauwels R., Baba M., Desmyter J., De Clercq E. Common features in the interaction of tetrahydroimidazo[4,5,1-jk][1,4]benzodiazepin-2(1H)-one and -thione and 1-[(2-hydroxyethoxy)methyl]-6-(phenylthio)thymine derivatives with the human immunodeficiency virus type 1 reverse transcriptase. Mol Pharmacol. 1992 May;41(5):963–968. [PubMed] [Google Scholar]
  14. Debyser Z., Vandamme A. M., Pauwels R., Baba M., Desmyter J., De Clercq E. Kinetics of inhibition of endogenous human immunodeficiency virus type 1 reverse transcription by 2',3'-dideoxynucleoside 5'-triphosphate, tetrahydroimidazo-[4,5,1-jk][1,4]-benzodiazepin-2(1H)-thion e, and 1-[(2-hydroxyethoxy)methyl]-6-(phenylthio)thymine derivatives. J Biol Chem. 1992 Jun 15;267(17):11769–11776. [PubMed] [Google Scholar]
  15. Goldman M. E., Nunberg J. H., O'Brien J. A., Quintero J. C., Schleif W. A., Freund K. F., Gaul S. L., Saari W. S., Wai J. S., Hoffman J. M. Pyridinone derivatives: specific human immunodeficiency virus type 1 reverse transcriptase inhibitors with antiviral activity. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6863–6867. doi: 10.1073/pnas.88.15.6863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Goldman M. E., O'Brien J. A., Ruffing T. L., Nunberg J. H., Schleif W. A., Quintero J. C., Siegl P. K., Hoffman J. M., Smith A. M., Emini E. A. L-696,229 specifically inhibits human immunodeficiency virus type 1 reverse transcriptase and possesses antiviral activity in vitro. Antimicrob Agents Chemother. 1992 May;36(5):1019–1023. doi: 10.1128/aac.36.5.1019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Koyanagi Y., O'Brien W. A., Zhao J. Q., Golde D. W., Gasson J. C., Chen I. S. Cytokines alter production of HIV-1 from primary mononuclear phagocytes. Science. 1988 Sep 23;241(4873):1673–1675. doi: 10.1126/science.241.4873.1673. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Larder B. A., Darby G., Richman D. D. HIV with reduced sensitivity to zidovudine (AZT) isolated during prolonged therapy. Science. 1989 Mar 31;243(4899):1731–1734. doi: 10.1126/science.2467383. [DOI] [PubMed] [Google Scholar]
  20. Larder B. A., Kemp S. D. Multiple mutations in HIV-1 reverse transcriptase confer high-level resistance to zidovudine (AZT). Science. 1989 Dec 1;246(4934):1155–1158. doi: 10.1126/science.2479983. [DOI] [PubMed] [Google Scholar]
  21. Merluzzi V. J., Hargrave K. D., Labadia M., Grozinger K., Skoog M., Wu J. C., Shih C. K., Eckner K., Hattox S., Adams J. Inhibition of HIV-1 replication by a nonnucleoside reverse transcriptase inhibitor. Science. 1990 Dec 7;250(4986):1411–1413. doi: 10.1126/science.1701568. [DOI] [PubMed] [Google Scholar]
  22. Miyasaka T., Tanaka H., Baba M., Hayakawa H., Walker R. T., Balzarini J., De Clercq E. A novel lead for specific anti-HIV-1 agents: 1-[(2-hydroxyethoxy)methyl]-6-(phenylthio)thymine. J Med Chem. 1989 Dec;32(12):2507–2509. doi: 10.1021/jm00132a002. [DOI] [PubMed] [Google Scholar]
  23. Nunberg J. H., Schleif W. A., Boots E. J., O'Brien J. A., Quintero J. C., Hoffman J. M., Emini E. A., Goldman M. E. Viral resistance to human immunodeficiency virus type 1-specific pyridinone reverse transcriptase inhibitors. J Virol. 1991 Sep;65(9):4887–4892. doi: 10.1128/jvi.65.9.4887-4892.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pauwels R., Balzarini J., Baba M., Snoeck R., Schols D., Herdewijn P., Desmyter J., De Clercq E. Rapid and automated tetrazolium-based colorimetric assay for the detection of anti-HIV compounds. J Virol Methods. 1988 Aug;20(4):309–321. doi: 10.1016/0166-0934(88)90134-6. [DOI] [PubMed] [Google Scholar]
  25. Pauwels R., De Clercq E., Desmyter J., Balzarini J., Goubau P., Herdewijn P., Vanderhaeghe H., Vandeputte M. Sensitive and rapid assay on MT-4 cells for detection of antiviral compounds against the AIDS virus. J Virol Methods. 1987 Jun;16(3):171–185. doi: 10.1016/0166-0934(87)90002-4. [DOI] [PubMed] [Google Scholar]
  26. Perno C. F., Yarchoan R., Cooney D. A., Hartman N. R., Gartner S., Popovic M., Hao Z., Gerrard T. L., Wilson Y. A., Johns D. G. Inhibition of human immunodeficiency virus (HIV-1/HTLV-IIIBa-L) replication in fresh and cultured human peripheral blood monocytes/macrophages by azidothymidine and related 2',3'-dideoxynucleosides. J Exp Med. 1988 Sep 1;168(3):1111–1125. doi: 10.1084/jem.168.3.1111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Richman D., Rosenthal A. S., Skoog M., Eckner R. J., Chou T. C., Sabo J. P., Merluzzi V. J. BI-RG-587 is active against zidovudine-resistant human immunodeficiency virus type 1 and synergistic with zidovudine. Antimicrob Agents Chemother. 1991 Feb;35(2):305–308. doi: 10.1128/aac.35.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Richman D., Shih C. K., Lowy I., Rose J., Prodanovich P., Goff S., Griffin J. Human immunodeficiency virus type 1 mutants resistant to nonnucleoside inhibitors of reverse transcriptase arise in tissue culture. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11241–11245. doi: 10.1073/pnas.88.24.11241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Romero D. L., Busso M., Tan C. K., Reusser F., Palmer J. R., Poppe S. M., Aristoff P. A., Downey K. M., So A. G., Resnick L. Nonnucleoside reverse transcriptase inhibitors that potently and specifically block human immunodeficiency virus type 1 replication. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8806–8810. doi: 10.1073/pnas.88.19.8806. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tramontano E., Cheng Y. C. HIV-1 reverse transcriptase inhibition by a dipyridodiazepinone derivative: BI-RG-587. Biochem Pharmacol. 1992 Mar 17;43(6):1371–1376. doi: 10.1016/0006-2952(92)90515-k. [DOI] [PubMed] [Google Scholar]
  31. Yuasa S., Sadakata Y., Takashima H., Sekiya K., Inouye N., Ubasawa M., Baba M. Selective and synergistic inhibition of human immunodeficiency virus type 1 reverse transcriptase by a non-nucleoside inhibitor, MKC-442. Mol Pharmacol. 1993 Oct;44(4):895–900. [PubMed] [Google Scholar]
  32. de Vreese K., Debyser Z., Vandamme A. M., Pauwels R., Desmyter J., de Clercq E., Anné J. Resistance of human immunodeficiency virus type 1 reverse transcriptase to TIBO derivatives induced by site-directed mutagenesis. Virology. 1992 Jun;188(2):900–904. doi: 10.1016/0042-6822(92)90550-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES