New dose-time relationships of folate antagonists to sustain inhibition of human lymphoblasts and leukemic cells in vitro.
In this study, three methods are utilized to analyze toxicity produced by methotrexate and the lipid-soluble antifolate, 2,4-diamino-5-(3',4'-dichlorophenyl)-6-methylpyrimidine, in human lymphoblasts (WIL-2) and leukemic cells. These methods detect increasingly severe metabolic damage; inhibition of deoxyuridine incorporation into DNA, the reversibility of inhibition of deoxyuridine incorporation by supplementation with formyltetrahydrofolate as Ca2+ leucovorin, and the ability of cells to form clones in soft agarose. The critical dose and exposure time for establishing and maintaining the metabolic toxicity of methotrexate is examined in detail. It is shown that, if an initial loading dose of methotrexate is of high enough concentration or is maintained for a sufficient period to achieve greater than 98% inhibition of deoxyuridine incorporation, this inhibition can be sustained by low concentrations of 2,4-diamino-5-(3',4'-dichlorophenyl)-6-methylpyrimidine or methotrexate. Concentrations of methotrexate or 2,4-diamino-5-(3',4'-dichlorophenyl)-6-methylpyrimidine that equal or exceed 0.1 microM are sufficient for maintenance of inhibition by an initial loading dose of methotrexate but escape from inhibition that occurs if lower levels of drug are used. The possible implications of these observations for in vivo protocols are discussed.
D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003857:Deoxyuridine / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005493:Folic Acid Antagonists / Q000008:administration & dosage* / Q000494:pharmacology; D006801:Humans; D002955:Leucovorin / Q000494:pharmacology; D007938:Leukemia / Q000188:drug therapy; D008727:Methotrexate / Q000008:administration & dosage* / Q000378:metabolism / Q000494:pharmacology; D011739:Pyrimethamine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology; D013997:Time Factors
D005493:Folic Acid Antagonists; C006308:metoprine; D002955:Leucovorin; D003857:Deoxyuridine; D008727:Methotrexate; D011739:Pyrimethamine