Intracellular metabolism of 5,10-dideazatetrahydrofolic acid in human leukemia cell lines.
5,10-Dideazatetrahydrofolic acid (DDATHF) is a new potent antitumor agent that specifically inhibits purine biosynthesis, primarily through inhibition of glycinamide ribonucleotide transformylase, the first of the tetrahydrofolate-requiring enzymes in the de novo synthesis pathway. DDATHF has been shown to be an excellent substrate for mouse liver folylpolyglutamate synthetase in vitro, suggesting that intracellular conversion to polyglutamates could play an important role in the action of this antifolate. In this report, metabolic studies of the 6R-diastereomer of DDATHF in the cultured human leukemia cell lines CCRF-CEM and HL-60 are presented. At both 1 and 10 microM (6R)-DDATHF was rapidly converted to polyglutamates in both cell lines. DDATHF(Glu)5 and DDATHF(Glu)6 were the main intracellular metabolites. After incubation in drug-free medium, (6R)-DDATHF polyglutamates were better retained intracellularly with increasing glutamate chain length. (6R)-DDATHF showed reduced cytotoxicity toward a folylpolyglutamate synthetase-deficient cell line, CCRF-CEM30/6 related to a dramatically diminished accumulation of polyglutamates. The activity of (6R)-DDATHF in CCRF-CEM30/6 cells was decreased after both short and prolonged exposures. These results suggest that polyglutamylation of (6R)-DDATHF not only represents a mechanism for trapping the drug inside the cells but also produces a more potent inhibitor of the target enzyme.
Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06510.
G
G
Pizzorno
J A
JA
Sokoloski
A R
AR
Cashmore
B A
BA
Moroson
A D
AD
Cross
G P
GP
Beardsley
D002455:Cell Division; D006801:Humans; D007945:Leukemia, Lymphoid / Q000378:metabolism*; D011099:Polyglutamic Acid / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D013763:Tetrahydrofolates / Q000737:chemistry / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000378:metabolism; D011623:gamma-Glutamyl Hydrolase / Q000378:metabolism
D013763:Tetrahydrofolates; D011099:Polyglutamic Acid; C045894:lometrexol; D011623:gamma-Glutamyl Hydrolase