In vitro metabolism of deoxycoformycin in human T lymphoblastoid cells. Phosphorylation of deoxycoformycin and incorporation into cellular DNA.
The biochemical and metabolic effects of deoxycoformycin, a potent inhibitor of adenosine deaminase, were investigated using two human T lymphoblastoid cell lines. A dose-response analysis demonstrated that the concentration of deoxycoformycin at which there was 50% inhibition of growth was greater than 1 X 10(-3) M in lymphoblastoid cells. Uptake of deoxycoformycin was biphasic and occurred much more slowly than for natural nucleosides, and lower saturation levels were reached. The intracellular concentration of deoxycoformycin achieved was 0.4 to 0.5 microM when the extracellular concentration was 1 microM. At 10 microM extracellular concentration, the intracellular concentration was 3-4 microM. Although deoxycoformycin at very low concentrations (1 or 10 microM) did not have any detectable effects on the growth of these cells, the nucleoside was found to be metabolized, and was phosphorylated to give the mono-, di-, and triphosphate derivatives. The triphosphate derivative was incorporated into cellular DNA with little incorporation into cellular RNA. Metabolism of deoxycoformycin in several mutant lymphoblastoid cells deficient in adenosine kinase and/or deoxycytidine kinase was found to be unchanged from wild-type cells, indicating that these major nucleoside kinases do not play a significant role in the phosphorylation of deoxycoformycin. These results may account, at least in part, for the differences that are observed between the pharmacologic inhibition of adenosine deaminase, and the inherited deficiency of adenosine deaminase.
D058892:Adenosine Deaminase Inhibitors; D000248:Adenosine Kinase / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003070:Coformycin / Q000031:analogs & derivatives / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D003839:Deoxyadenosines / Q000378:metabolism; D003842:Deoxycytidine Kinase / Q000378:metabolism; D006801:Humans; D009700:Nucleoside Deaminases / Q000037:antagonists & inhibitors*; D015649:Pentostatin / Q000031:analogs & derivatives; D010766:Phosphorylation; D012313:RNA / Q000378:metabolism; D012263:Ribonucleosides / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism*; D014740:Vidarabine / Q000494:pharmacology
D058892:Adenosine Deaminase Inhibitors; D003838:Deoxyadenine Nucleotides; D003839:Deoxyadenosines; D012263:Ribonucleosides; D003070:Coformycin; D015649:Pentostatin; D012313:RNA; C022803:2'-deoxycoformycin 5'-phosphate; D004247:DNA; D000248:Adenosine Kinase; D003842:Deoxycytidine Kinase; D009700:Nucleoside Deaminases; D014740:Vidarabine; C026600:2'-deoxyadenosine triphosphate