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. 1997 May;41(5):1042–1045. doi: 10.1128/aac.41.5.1042

Efficacy of trimethoprim in murine experimental infection with a thymidine kinase-deficient mutant of Escherichia coli.

T Tokunaga 1, K Oka 1, A Takemoto 1, Y Ohtsubo 1, N Gotoh 1, T Nishino 1
PMCID: PMC163847  PMID: 9145866

Abstract

The antimicrobial activity of trimethoprim is antagonized by thymidine in in vitro susceptibility tests. The purpose of this investigation was to determine whether this antagonism also occurred during experimental infection in mice, which have high serum thymidine concentrations. We derived a mutant strain of Escherichia coli, TT-48, incapable of utilizing exogenous thymidine from parent strain E. coli KC-14 and then investigated the in vitro and in vivo antimicrobial activities of trimethoprim, sulfamethoxazole, cefdinir, and ofloxacin against these strains. E. coli TT-48 lacked the activity of thymidine kinase, which catalyzes the conversion of thymidine to thymidylate, but its growth curve remained close to that of the parent strain. The MICs of all of the antimicrobial agents tested, except cefdinir, for the mutant strain were slightly inferior to those for the parent strain. The bactericidal effect of trimethoprim against the parent strain was antagonized by thymidine at concentrations of more than 1 microg/ml, while that against the mutant strain was not affected by thymidine even at the highest concentration (10 microg/ml). The therapeutic efficacy of trimethoprim in experimental murine infections was significantly higher when the mutant rather than the parent strain was used, whereas the therapeutic efficacy of cefdinir or ofloxacin, whose antimicrobial action is independent of folic acid synthesis, was the same with both strains. Unexpectedly, sulfamethoxazole also had similar efficacy against both strains. Thus, high thymidine concentrations antagonized the antimicrobial activity of trimethoprim in vitro and in vivo.

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Selected References

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  1. Burchall J. J., Hitchings G. H. Inhibitor binding analysis of dihydrofolate reductases from various species. Mol Pharmacol. 1965 Sep;1(2):126–136. [PubMed] [Google Scholar]
  2. FURLONG N. B. A rapid assay for nucleotide kinases using C14-or H3-labeled nucleotides. Anal Biochem. 1963 Jun;5:515–522. doi: 10.1016/0003-2697(63)90071-x. [DOI] [PubMed] [Google Scholar]
  3. Grunberg E., DeLorenzo W. F., Titsworth E. Demonstration of the therapeutic effectiveness of sulfamethoxazole in experimental infections of mice. Chemotherapy. 1970;15(2):99–104. doi: 10.1159/000220671. [DOI] [PubMed] [Google Scholar]
  4. Hamilton-Miller J. M. Reversal of activity of trimethoprim against gram-positive cocci by thymidine, thymine and 'folates'. J Antimicrob Chemother. 1988 Jul;22(1):35–39. doi: 10.1093/jac/22.1.35. [DOI] [PubMed] [Google Scholar]
  5. Hiraga S., Igarashi K., Yura T. A deoxythymidine kinase-deficient mutant of Escherichia coli. I. Isolation and some properties. Biochim Biophys Acta. 1967 Aug 22;145(1):41–51. doi: 10.1016/0005-2787(67)90652-1. [DOI] [PubMed] [Google Scholar]
  6. Hitchings G. H., Burchall J. J. Inhibition of folate biosynthesis and function as a basis for chemotherapy. Adv Enzymol Relat Areas Mol Biol. 1965;27:417–468. doi: 10.1002/9780470122723.ch9. [DOI] [PubMed] [Google Scholar]
  7. Hitchings G. H. Species differences among dihydrofolate reductases as a basis for chemotherapy. Postgrad Med J. 1969 Nov;45(Suppl):7–10. [PubMed] [Google Scholar]
  8. KRENITSKY T. A., BARCLAY M., JACQUEZ J. A. SPECIFICITY OF MOUSE URIDINE PHOSPHORYLASE. CHROMATOGRAPHY, PURIFICATION, AND PROPERTIES. J Biol Chem. 1964 Mar;239:805–812. [PubMed] [Google Scholar]
  9. Koch A. E., Burchall J. J. Reversal of the antimicrobial activity of trimethoprim by thymidine in commercially prepared media. Appl Microbiol. 1971 Nov;22(5):812–817. doi: 10.1128/am.22.5.812-817.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Meshi T., Sato Y. Studies on sulfamethoxazole-trimethoprim. Absorption, distribution, excretion and metabolism of trimethoprim in rat. Chem Pharm Bull (Tokyo) 1972 Oct;20(10):2079–2090. doi: 10.1248/cpb.20.2079. [DOI] [PubMed] [Google Scholar]
  12. Nottebrock H., Then R. Thymidine concentrations in serum and urine of different animal species and man. Biochem Pharmacol. 1977 Nov 15;26(22):2175–2179. doi: 10.1016/0006-2952(77)90271-4. [DOI] [PubMed] [Google Scholar]
  13. Schwartz D. E., Rieder J. Pharmacokinetics of sulfamethoxazole plus trimethoprim in man and their distribution in the rat. Chemotherapy. 1970;15(6):337–355. doi: 10.1159/000220701. [DOI] [PubMed] [Google Scholar]
  14. Schwartz D. E., Vetter W., Englert G. Trimethoprim metabolites in rat, dog and man: qualitative and quantitative studies. Arzneimittelforschung. 1970 Dec;20(12):1867–1871. [PubMed] [Google Scholar]
  15. Stokes A., Lacey R. W. Effect of thymidine on activity of trimethoprim and sulphamethoxazole. J Clin Pathol. 1978 Feb;31(2):165–171. doi: 10.1136/jcp.31.2.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sugino A., Peebles C. L., Kreuzer K. N., Cozzarelli N. R. Mechanism of action of nalidixic acid: purification of Escherichia coli nalA gene product and its relationship to DNA gyrase and a novel nicking-closing enzyme. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4767–4771. doi: 10.1073/pnas.74.11.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Then R., Angehrn P. The biochemical basis of the antimicrobial action of sulfonamides and trimethoprim in vivo--I. Action of sulfonamides and trimethoprim in blood and urine. Biochem Pharmacol. 1974 Nov 1;23(21):2977–2982. doi: 10.1016/0006-2952(74)90272-x. [DOI] [PubMed] [Google Scholar]
  18. Tomasz A. The mechanism of the irreversible antimicrobial effects of penicillins: how the beta-lactam antibiotics kill and lyse bacteria. Annu Rev Microbiol. 1979;33:113–137. doi: 10.1146/annurev.mi.33.100179.000553. [DOI] [PubMed] [Google Scholar]
  19. Wachsman J. T., Kemp S., Hogg L. Comparative effects of 5-fluorouracil on strains of Bacillus megaterium. J Bacteriol. 1964 May;87(5):1011–1018. doi: 10.1128/jb.87.5.1011-1018.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]

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