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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1975 Oct;72(10):4018–4022. doi: 10.1073/pnas.72.10.4018

New approach to antifolate treatment of certain cancers as demonstrated in tissue culture.

R M Halpern, B C Halpern, B R Clark, H Ashe, D N Hardy, P Y Jenkinson, S C Chou, R A Smith
PMCID: PMC433129  PMID: 1081693

Abstract

The selective toxicity of antifolates for a variety of cancers can be improved, as illustrated by the combined administration of N5-methyltetrahydrofolate and methotrexate in tissue culture. When a variety of neoplastic cell types characterized by a deficiency of vitamin B12-dependent N5-methyltetrahydrofolate methyltransferase (5-methyltetrahydropteroyl-L-glutamate:L-homocysteine S-methyltransferase, EC 2.1.1.13) and normal adult cells are grown in media containing methotrexate and either N5-methyltetrahydrofolate or N5-formyltetrahydrofolate, not only is the selective toxicity of methotrexate demonstrated, but the advantage of using N5-methyltetrahydrofolate in place of N5-formyltetrahydrofolate is also revealed. The implications and applications of this particular combination in the treatment of human cancer are discussed.

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Selected References

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

  1. Ashe H., Clark B. R., Chu F., Hardy D. N., Halpern B. C., Halpern R. M., Smith R. A. N5-methyltetrahydrofolate: homocysteine methyltransferase activity in extracts from normal, malignant and embryonic tissue culture cells. Biochem Biophys Res Commun. 1974 Mar 25;57(2):417–425. doi: 10.1016/0006-291x(74)90947-4. [DOI] [PubMed] [Google Scholar]
  2. BERTINO J. R. THE MECHANISM OF ACTION OF THE FOLATE ANTAGONISTS IN MAN. Cancer Res. 1963 Sep;23:1286–1306. [PubMed] [Google Scholar]
  3. BURCHENAL J. H., BABCOCK G. M., BROQUIST H. P., JUKES T. H. Prevention of chemotherapeutic effects of 4-amino-N10-methyl-pteroylglutamic acid on mouse leukemia by citrovorum factor. Proc Soc Exp Biol Med. 1950 Aug;74(4):735–737. doi: 10.3181/00379727-74-18031. [DOI] [PubMed] [Google Scholar]
  4. Baker B. R., Lourens G. J. Irreversible enzyme inhibitors. CV. Differential irreversible inhibition of vertebrate dihydrofolic reductases by derivatives of 4,6-diamino-1,2-dihydro-2,2-dimethyl-1-phenyl-s-triazines substituted with a terminal sulfonyl fluoride. J Med Chem. 1967 Nov;10(6):1113–1122. doi: 10.1021/jm00318a029. [DOI] [PubMed] [Google Scholar]
  5. Borsa J., Whitmore G. F. Studies relating to the mode of action of methotrexate. II. Studies on sites of action in L-cells in vitro. Mol Pharmacol. 1969 Jul;5(4):303–317. [PubMed] [Google Scholar]
  6. Bruce W. R., Meeker B. E., Valeriote F. A. Comparison of the sensitivity of normal hematopoietic and transplanted lymphoma colony-forming cells to chemotherapeutic agents administered in vivo. J Natl Cancer Inst. 1966 Aug;37(2):233–245. [PubMed] [Google Scholar]
  7. Bruce W. R., Meeker B. E., Valeriote F. A. Comparison of the sensitivity of normal hematopoietic and transplanted lymphoma colony-forming cells to chemotherapeutic agents administered in vivo. J Natl Cancer Inst. 1966 Aug;37(2):233–245. [PubMed] [Google Scholar]
  8. Clark B. R., Ashe H., Halpern R. M., Smith R. A. A method for determination of methionine containing radioactivity in the thiomethyl moiety. Anal Biochem. 1974 Sep;61(1):243–247. doi: 10.1016/0003-2697(74)90351-0. [DOI] [PubMed] [Google Scholar]
  9. Cohen S. S. On the nature of thymineless death. Ann N Y Acad Sci. 1971 Nov 30;186:292–301. doi: 10.1111/j.1749-6632.1971.tb31155.x. [DOI] [PubMed] [Google Scholar]
  10. Djerassi I., Rominger C. J., Kim J. S., Turchi J., Suvansri U., Hughes D. Phase I study of high doses of methotrexate with citrovorum factor in patients with lung cancer. Cancer. 1972 Jul;30(1):22–30. doi: 10.1002/1097-0142(197207)30:1<22::aid-cncr2820300105>3.0.co;2-b. [DOI] [PubMed] [Google Scholar]
  11. FUTTERMAN S. Enzymatic reduction of folic acid and dihydrofolic acid to tetrahydrofolic acid. J Biol Chem. 1957 Oct;228(2):1031–1038. [PubMed] [Google Scholar]
  12. GOLDIN A., HUMPHREYS S. R., MANTEL N., VENDITTI J. M. Modification of treatment of schedules in the management of advanced mouse leukemia with amethopterin. J Natl Cancer Inst. 1956 Aug;17(2):203–212. [PubMed] [Google Scholar]
  13. Goldman I. D. The characteristics of the membrane transport of amethopterin and the naturally occurring folates. Ann N Y Acad Sci. 1971 Nov 30;186:400–422. doi: 10.1111/j.1749-6632.1971.tb46996.x. [DOI] [PubMed] [Google Scholar]
  14. HATCH F. T., LARRABEE A. R., CATHOU R. E., BUCHANAN J. M. Enzymatic synthesis of the methyl group of methionine. I. Identification of the enzymes and cofactors involved in the system isolated from Escherichia coli. J Biol Chem. 1961 Apr;236:1095–1101. [PubMed] [Google Scholar]
  15. HERBERT V., ZALUSKY R. Interrelations of vitamin B12 and folic acid metabolism: folic acid clearance studies. J Clin Invest. 1962 Jun;41:1263–1276. doi: 10.1172/JCI104589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. HOLLAND J. F. Symposium on the experimental pharmacology and clinical use of antimetabolites. VII. Folic acid antagonists. Clin Pharmacol Ther. 1961 May-Jun;2:374–409. doi: 10.1002/cpt196123374. [DOI] [PubMed] [Google Scholar]
  17. Halpern B. C., Clark B. R., Hardy D. N., Halpern R. M., Smith R. A. The effect of replacement of methionine by homocystine on survival of malignant and normal adult mammalian cells in culture. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1133–1136. doi: 10.1073/pnas.71.4.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Halpern B. C., Ezzell R., Hardy D. N., Clark B. R., Ashe H., Halpern R. M., Smith R. A. Effect of methionine replacement by homocystine in cultures containing both malignant rat breast carcinosarcoma (Walker-256) cells and normal adult rat liver fibroblasts. In Vitro. 1975 Jan-Feb;11(1):14–19. doi: 10.1007/BF02615317. [DOI] [PubMed] [Google Scholar]
  19. Hryniuk W. M., Bertino J. R. Growth rate and cell kill. Ann N Y Acad Sci. 1971 Nov 30;186:330–342. [PubMed] [Google Scholar]
  20. JUKES T. H., FRANKLIN A. L., STOKSTAD E. L. R. Pteroylglutamic acid antagonists. Ann N Y Acad Sci. 1950 Jul 7;52(8):1336–1341. doi: 10.1111/j.1749-6632.1950.tb54033.x. [DOI] [PubMed] [Google Scholar]
  21. Jaffe N., Paed D., Farber S., Traggis D., Geiser C., Kim B. S., Das L., Frauenberger G., Djerassi I., Cassady J. R. Favorable response of metastatic osteogenic sarcoma to pulse high-dose methotrexate with citrovorum rescue and radiation therapy. Cancer. 1973 Jun;31(6):1367–1373. doi: 10.1002/1097-0142(197306)31:6<1367::aid-cncr2820310611>3.0.co;2-6. [DOI] [PubMed] [Google Scholar]
  22. KATZEN H. M., BUCHANAN J. M. ENZYMATIC SYNTHESIS OF THE METHYL GROUP OF METHIONINE. 8. REPRESSION-DEREPRESSION, PURIFICATION, AND PROPERTIES OF 5,10-METHYLENETETRAHYDROFOLATE REDUCTASE FROM ESCHERICHIA COLI. J Biol Chem. 1965 Feb;240:825–835. [PubMed] [Google Scholar]
  23. KISLIUK R. L. The source of hydrogen for methionine methyl formation. J Biol Chem. 1963 Jan;238:397–400. [PubMed] [Google Scholar]
  24. Kamely D., Littlefield J. W., Erbe R. W. Regulation of 5-methyltetrahydrofolate: homocysteine methyltransferase activity by methionine, vitamin B12, and folate in cultured baby hamster kidney cells. Proc Natl Acad Sci U S A. 1973 Sep;70(9):2585–2589. doi: 10.1073/pnas.70.9.2585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kessel D., Hall T. C., Roberts D. Modes of uptake of methotrexate by normal and leukemic human leukocytes in vitro and their relation to drug response. Cancer Res. 1968 Mar;28(3):564–570. [PubMed] [Google Scholar]
  26. Kutzbach C., Stokstad E. L. Mammalian methylenetetrahydrofolate reductase. Partial purification, properties, and inhibition by S-adenosylmethionine. Biochim Biophys Acta. 1971 Dec 15;250(3):459–477. doi: 10.1016/0005-2744(71)90247-6. [DOI] [PubMed] [Google Scholar]
  27. Metz J., Kelly A., Swett V. C., Waxman S., Herbert V. Deranged DNA synthesis by bone marrow from vitamin B-12-deficient humans. Br J Haematol. 1968 Jun;14(6):575–592. doi: 10.1111/j.1365-2141.1968.tb00364.x. [DOI] [PubMed] [Google Scholar]
  28. Mudd S. H., Uhlendorf B. W., Hinds K. R. Deranged B 12 metabolism: studies of fibroblasts grown in tissue culture. Biochem Med. 1970 Nov;4(3):215–239. doi: 10.1016/0006-2944(70)90050-5. [DOI] [PubMed] [Google Scholar]
  29. Nixon P. F., Bertino J. R. Impaired utilization of serum folate in pernicious anemia. A study with radiolabeled 5-methyltetrahydrofolate. J Clin Invest. 1972 Jun;51(6):1431–1439. doi: 10.1172/JCI106939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. OSBORN M. J., FREEMAN M., HUENNEKENS F. M. Inhibition of dihydrofolic reductase by aminopterin and amethopterin. Proc Soc Exp Biol Med. 1958 Feb;97(2):429–431. doi: 10.3181/00379727-97-23764. [DOI] [PubMed] [Google Scholar]
  31. PHILIPS F. S., THIERSCH J. B., FERGUSON F. C. Studies of the action of 4-aminopteroylglutamic acid and its congeners in mammals. Ann N Y Acad Sci. 1950 Jul 7;52(8):1349–1359. doi: 10.1111/j.1749-6632.1950.tb54036.x. [DOI] [PubMed] [Google Scholar]
  32. SCHOENBACH E. B., COLSKY J., GREENSPAN E. M. Observations on the effects of the folic acid antagonists, aminopterin and amethopterin, in patients with advanced neoplasms. Cancer. 1952 Nov;5(6):1201–1220. doi: 10.1002/1097-0142(195211)5:6<1201::aid-cncr2820050617>3.0.co;2-1. [DOI] [PubMed] [Google Scholar]
  33. SILVERMAN M., KERESZTESY J. C., KOVAL G. J., GARDINER R. C. Citrovorum factor and the synthesis of formylglutamic acid. J Biol Chem. 1957 May;226(1):83–94. [PubMed] [Google Scholar]
  34. SKIPPER H. E., MITCHELL J. H., Jr, BENNETT L. L., Jr Inhibition of nucleic acid synthesis by folic acid antagonists. Cancer Res. 1950 Aug;10(8):510–512. [PubMed] [Google Scholar]
  35. SKIPPER H. E., SCHABEL F. M., Jr, BELL M., THOMSON J. R., JOHNSON S. On the curability of experimental neoplasms. I. Amethopterin and mouse leukemias. Cancer Res. 1957 Aug;17(7):717–726. [PubMed] [Google Scholar]
  36. STEVENS A., SAKAMI W. Biosynthesis of methionine in liver. J Biol Chem. 1959 Aug;234(8):2063–2072. [PubMed] [Google Scholar]
  37. SULLIVAN R. D., MILLER E., SIKES M. P. Antimetabolite-metabolite combination cancer chemotherapy. Effects of intraarterial methotrexate-intramuscular Citrovorum factor therapy in human cancer. Cancer. 1959 Nov-Dec;12:1248–1262. doi: 10.1002/1097-0142(195911/12)12:6<1248::aid-cncr2820120619>3.0.co;2-2. [DOI] [PubMed] [Google Scholar]
  38. SULLIVAN R. D., WOOD A. M., CLIFFORD P., DUFF J. K., TRUSSEL R., NARY D. K., BURCHENAL J. H. Continuous intraarterial methotrexate with simultaneous, intermittent, intramuscular citrovorum factor herapy in carcinoma of the cervix. Cancer Chemother Rep. 1960 Jul;8:1–6. [PubMed] [Google Scholar]
  39. TABOR H., WYNGARDEN L. The enzymatic formation of formiminotetrahydrofolic acid, 5,10-methenyltetrahydrofolic acid, and 10-formyltetrahydrofolic acid in the metabolism of formiminoglutamic acid. J Biol Chem. 1959 Jul;234(7):1830–1846. [PubMed] [Google Scholar]
  40. WEISSBACH H., PETERKOFSKY A., REDFIELD B. G., DICKERMAN H. STUDIES ON THE TERMINAL REACTION IN THE BIOSYNTHESIS OF METHIONINE. J Biol Chem. 1963 Oct;238:3318–3324. [PubMed] [Google Scholar]
  41. WERKHEISER W. C. THE BIOCHEMICAL, CELLULAR, AND PHARMACOLOGICAL ACTION AND EFFECTS OF THE FOLIC ACID ANTAGONISTS. Cancer Res. 1963 Sep;23:1277–1285. [PubMed] [Google Scholar]
  42. ZAKRZEWSKI S. F., NICHOL C. A. On the enzymic reduction of folic acid by a purified hydrogenase. Biochim Biophys Acta. 1958 Feb;27(2):425–426. doi: 10.1016/0006-3002(58)90360-3. [DOI] [PubMed] [Google Scholar]

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