Skip to main content
NCBI home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1993 Mar;37(3):580–591. doi: 10.1128/aac.37.3.580

Mevinolin (lovastatin) potentiates the antiproliferative effects of ketoconazole and terbinafine against Trypanosoma (Schizotrypanum) cruzi: in vitro and in vivo studies.

J A Urbina 1, K Lazardi 1, E Marchan 1, G Visbal 1, T Aguirre 1, M M Piras 1, R Piras 1, R A Maldonado 1, G Payares 1, W de Souza 1
PMCID: PMC187710  PMID: 8460926

Abstract

We have studied the antiproliferative effects of mevinolin (lovastatin), an inhibitor of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase, on the protozoan parasite Trypanosoma (Schizotrypanum) cruzi and its ability to potentiate the action of specific ergosterol biosynthesis inhibitors, such as ketoconazole and terbinafine, both in vitro and in vivo. Against the epimastigote form in vitro, mevinolin produced a dose-dependent reduction of the growth rate up to 25 microM, but at 50 and 75 microM, complete growth arrest and cell lysis took place after 144 and 96 h, respectively. A systematic study of the effects of mevinolin combined with ketoconazole and terbinafine, which act at different points in the ergosterol biosynthesis pathway, on the proliferation of epimastigotes indicated a synergic action, as shown by concave isobolograms and fractional inhibitory concentration indexes ranging from 0.17 to 0.54. Analysis of the sterol composition and de novo sterol synthesis in control and treated cells by thin-layer and gas-liquid chromatographies showed that the antiproliferative effects of the drug alone and in combination were correlated with the depletion of the endogenous ergosterol pool and particularly with a critical (exogenous) cholesterol/endogenous 4-desmethyl sterol ratio in the cells. When we studied the effects of mevinolin on the amastigote form proliferating inside Vero cells in vitro, only very modest effects on the parasites were observed up to 0.75 microM; above this concentration, significant deleterious effects on the host cells were found. However, when the same concentration of the drug was combined with ketoconazole, it was able to reduce by a factor of 10 the concentration of the azole required to eradicate the parasite (from 10 to 1 nM), again indicating a synergic action. On the other hand, a combination of mevinolin and terbinafine had only additive effects on amastigotes, but a ternary combination of mevinolin, ketoconazole, and terbinafine was again clearly synergistic. In vivo studies with a murine model of Chagas' disease showed that mevinolin can also potentiate the therapeutic effects of ketoconazole in this system; combined treatment with the two drugs at doses that alone offered only limited protection against the parasite was able to essentially eliminate circulating parasites and produce complete protection against death. These results confirm the synergic action against the proliferative stages of T. cruzi both in vitro and in vivo and in vivo of combined ergosterol biosynthesis inhibitors that act at different points in the pathway and suggest that mevinolin combined with azoles, such as ketoconazole, can be used in the treatment of human Chagas' disease.

Full text

PDF
580

Selected References

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

  1. Alberts A. W., Chen J., Kuron G., Hunt V., Huff J., Hoffman C., Rothrock J., Lopez M., Joshua H., Harris E. Mevinolin: a highly potent competitive inhibitor of hydroxymethylglutaryl-coenzyme A reductase and a cholesterol-lowering agent. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3957–3961. doi: 10.1073/pnas.77.7.3957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beach D. H., Goad L. J., Holz G. G., Jr Effects of antimycotic azoles on growth and sterol biosynthesis of Leishmania promastigotes. Mol Biochem Parasitol. 1988 Nov;31(2):149–162. doi: 10.1016/0166-6851(88)90166-1. [DOI] [PubMed] [Google Scholar]
  3. Beach D. H., Goad L. J., Holz G. G., Jr Effects of ketoconazole on sterol biosynthesis by Trypanosoma cruzi epimastigotes. Biochem Biophys Res Commun. 1986 May 14;136(3):851–856. doi: 10.1016/0006-291x(86)90410-9. [DOI] [PubMed] [Google Scholar]
  4. Berman J. D. Activity of imidazoles against Leishmania tropica in human macrophage cultures. Am J Trop Med Hyg. 1981 May;30(3):566–569. doi: 10.4269/ajtmh.1981.30.566. [DOI] [PubMed] [Google Scholar]
  5. Berman J. D., Goad L. J., Beach D. H., Holz G. G., Jr Effects of ketoconazole on sterol biosynthesis by Leishmania mexicana mexicana amastigotes in murine macrophage tumor cells. Mol Biochem Parasitol. 1986 Jul;20(1):85–92. doi: 10.1016/0166-6851(86)90145-3. [DOI] [PubMed] [Google Scholar]
  6. Berman J. D., Holz G. G., Jr, Beach D. H. Effects of ketoconazole on growth and sterol biosynthesis of Leishmania mexicana promastigotes in culture. Mol Biochem Parasitol. 1984 May;12(1):1–13. doi: 10.1016/0166-6851(84)90039-2. [DOI] [PubMed] [Google Scholar]
  7. Borelli D. A clinical trial of itraconazole in the treatment of deep mycoses and leishmaniasis. Rev Infect Dis. 1987 Jan-Feb;9 (Suppl 1):S57–S63. doi: 10.1093/clinids/9.supplement_1.s57. [DOI] [PubMed] [Google Scholar]
  8. Docampo R., Moreno S. N., Turrens J. F., Katzin A. M., Gonzalez-Cappa S. M., Stoppani A. O. Biochemical and ultrastructural alterations produced by miconazole and econazole in Trypanosoma cruzi. Mol Biochem Parasitol. 1981 Jul;3(3):169–180. doi: 10.1016/0166-6851(81)90047-5. [DOI] [PubMed] [Google Scholar]
  9. Endo A. Chemistry, biochemistry, and pharmacology of HMG-CoA reductase inhibitors. Klin Wochenschr. 1988 May 16;66(10):421–427. doi: 10.1007/BF01745510. [DOI] [PubMed] [Google Scholar]
  10. Endo A. Monacolin K, a new hypocholesterolemic agent produced by a Monascus species. J Antibiot (Tokyo) 1979 Aug;32(8):852–854. doi: 10.7164/antibiotics.32.852. [DOI] [PubMed] [Google Scholar]
  11. Florin-Christensen M., Florin-Christensen J., Garin C., Isola E., Brenner R. R., Rasmussen L. Inhibition of Trypanosoma cruzi growth and sterol biosynthesis by lovastatin. Biochem Biophys Res Commun. 1990 Feb 14;166(3):1441–1445. doi: 10.1016/0006-291x(90)91028-q. [DOI] [PubMed] [Google Scholar]
  12. Goad L. J., Berens R. L., Marr J. J., Beach D. H., Holz G. G., Jr The activity of ketoconazole and other azoles against Trypanosoma cruzi: biochemistry and chemotherapeutic action in vitro. Mol Biochem Parasitol. 1989 Jan 15;32(2-3):179–189. doi: 10.1016/0166-6851(89)90069-8. [DOI] [PubMed] [Google Scholar]
  13. Goad L. J., Holz G. G., Jr, Beach D. H. Sterols of ketoconazole-inhibited Leishmania mexicana mexicana promastigotes. Mol Biochem Parasitol. 1985 Jun;15(3):257–279. doi: 10.1016/0166-6851(85)90089-1. [DOI] [PubMed] [Google Scholar]
  14. Hallander H. O., Dornbusch K., Gezelius L., Jacobson K., Karlsson I. Synergism between aminoglycosides and cephalosporins with antipseudomonal activity: interaction index and killing curve method. Antimicrob Agents Chemother. 1982 Nov;22(5):743–752. doi: 10.1128/aac.22.5.743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hart D. T., Lauwers W. J., Willemsens G., Vanden Bossche H., Opperdoes F. R. Perturbation of sterol biosynthesis by itraconazole and ketoconazole in Leishmania mexicana mexicana infected macrophages. Mol Biochem Parasitol. 1989 Mar 1;33(2):123–134. doi: 10.1016/0166-6851(89)90026-1. [DOI] [PubMed] [Google Scholar]
  16. Ikeura R., Murakawa S., Endo A. Growth inhibition of yeast by compactin (ML-236B) analogues. J Antibiot (Tokyo) 1988 Aug;41(8):1148–1150. doi: 10.7164/antibiotics.41.1148. [DOI] [PubMed] [Google Scholar]
  17. Larralde G., Vivas J., Urbina J. A. Concentration and time dependence of the effects of ketoconazole on growth and sterol synthesis by Trypanosoma (Schizotrypanum) cruzi epimastigotes. Acta Cient Venez. 1988;39(2):140–146. [PubMed] [Google Scholar]
  18. Lazardi K., Urbina J. A., de Souza W. Ultrastructural alterations induced by ICI 195,739, a bis-triazole derivative with strong antiproliferative action against Trypanosoma (Schizotrypanum) cruzi. Antimicrob Agents Chemother. 1991 Apr;35(4):736–740. doi: 10.1128/aac.35.4.736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lazardi K., Urbina J. A., de Souza W. Ultrastructural alterations induced by two ergosterol biosynthesis inhibitors, ketoconazole and terbinafine, on epimastigotes and amastigotes of Trypanosoma (Schizotrypanum) cruzi. Antimicrob Agents Chemother. 1990 Nov;34(11):2097–2105. doi: 10.1128/aac.34.11.2097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lorenz R. T., Parks L. W. Effects of lovastatin (mevinolin) on sterol levels and on activity of azoles in Saccharomyces cerevisiae. Antimicrob Agents Chemother. 1990 Sep;34(9):1660–1665. doi: 10.1128/aac.34.9.1660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Marr J. J., Docampo R. Chemotherapy for Chagas' disease: a perspective of current therapy and considerations for future research. Rev Infect Dis. 1986 Nov-Dec;8(6):884–903. doi: 10.1093/clinids/8.6.884. [DOI] [PubMed] [Google Scholar]
  22. McCabe R. E., Remington J. S., Araujo F. G. In vitro and in vivo effects of itraconazole against Trypanosoma cruzi. Am J Trop Med Hyg. 1986 Mar;35(2):280–284. doi: 10.4269/ajtmh.1986.35.280. [DOI] [PubMed] [Google Scholar]
  23. McCabe R. E., Remington J. S., Araujo F. G. Ketoconazole inhibition of intracellular multiplication of Trypanosoma cruzi and protection of mice against lethal infection with the organism. J Infect Dis. 1984 Oct;150(4):594–601. doi: 10.1093/infdis/150.4.594. [DOI] [PubMed] [Google Scholar]
  24. McCabe R. E., Remington J. S., Araujo F. G. Ketoconazole promotes parasitological cure of mice infected with Trypanosoma cruzi. Trans R Soc Trop Med Hyg. 1987;81(4):613–615. doi: 10.1016/0035-9203(87)90430-5. [DOI] [PubMed] [Google Scholar]
  25. Raether W., Seidenath H. Ketoconazole and other potent antimycotic azoles exhibit pronounced activity against Trypanosoma cruzi, Plasmodium berghei and Entamoeba histolytica in vivo. Z Parasitenkd. 1984;70(1):135–138. doi: 10.1007/BF00929583. [DOI] [PubMed] [Google Scholar]
  26. Stütz A. Synthesis and structure-activity correlations within allylamine antimycotics. Ann N Y Acad Sci. 1988;544:46–62. doi: 10.1111/j.1749-6632.1988.tb40388.x. [DOI] [PubMed] [Google Scholar]
  27. Urbina J. A., Lazardi K., Aguirre T., Piras M. M., Piras R. Antiproliferative effects and mechanism of action of ICI 195,739, a novel bis-triazole derivative, on epimastigotes and amastigotes of Trypanosoma (Schizotrypanum) cruzi. Antimicrob Agents Chemother. 1991 Apr;35(4):730–735. doi: 10.1128/aac.35.4.730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Urbina J. A., Lazardi K., Aguirre T., Piras M. M., Piras R. Antiproliferative synergism of the allylamine SF 86-327 and ketoconazole on epimastigotes and amastigotes of Trypanosoma (Schizotrypanum) cruzi. Antimicrob Agents Chemother. 1988 Aug;32(8):1237–1242. doi: 10.1128/aac.32.8.1237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Urbina J. A., Vivas J., Ramos H., Larralde G., Aguilar Z., Avilán L. Alteration of lipid order profile and permeability of plasma membranes from Trypanosoma cruzi epimastigotes grown in the presence of ketoconazole. Mol Biochem Parasitol. 1988 Aug;30(2):185–195. doi: 10.1016/0166-6851(88)90111-9. [DOI] [PubMed] [Google Scholar]
  30. Urcuyo F. G., Zaias N. Oral ketoconazole in the treatment of leishmaniasis. Int J Dermatol. 1982 Sep;21(7):414–416. doi: 10.1111/j.1365-4362.1982.tb03163.x. [DOI] [PubMed] [Google Scholar]
  31. Vanden Bossche H., Marichal P., Gorrens J., Geerts H., Janssen P. A. Mode of action studies. Basis for the search of new antifungal drugs. Ann N Y Acad Sci. 1988;544:191–207. doi: 10.1111/j.1749-6632.1988.tb40404.x. [DOI] [PubMed] [Google Scholar]
  32. de Maio A., Urbina J. A. Trypanosoma (Schizotrypanum) cruzi: terminal oxidases in exponential and stationary growth phase emipastigotes cultured in vitro. Acta Cient Venez. 1984;35(2):136–141. [PubMed] [Google Scholar]

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

RESOURCES