Structural specificity of inhibition of human folylpolyglutamate synthetase by ornithine-containing folate analogs.
A series of folate analogs containing ornithine instead of glutamate was synthesized and tested for inhibition of folylpolyglutamate synthetase (FPGS) and other folate-dependent enzymes of human leukemia cell lines. Reduced derivatives of 2-amino-4-oxo-10-methyl-pteroyl-ornithine had dramatically increased inhibitory potency against FPGS compared to the oxidized parent. The amino-pterin analog (2,4-diamino-pteroylornithine) was a potent inhibitor of both dihydrofolate reductase and FPGS. It was a much more potent linear competitive inhibitor of human FPGS than the corresponding methotrexate derivative previously described (Ki = 0.15-0.26 and 3 microM respectively). A quinazoline folate analog, 2-amino-4-oxo-5,8-dideazapteroyl-ornithine, was a relatively poor inhibitor of isolated dihydrofolate reductase and thymidylate synthase; however, it is the most potent human FPGS inhibitor identified to date (Ki = 100-150 nM). Because of the lack of appreciable interaction with other folate-dependent enzymes, structures incorporating the 2-amino-4-oxo-5,8-dideazapteroate nucleus may thus lead to selective inhibition of FPGS. Substitution of ornithine for glutamate caused a profound decrease in cytotoxic potency for these analogs; this was apparently the result of poor transport. Together with earlier studies, these data indicate that the potency of FPGS inhibition by an analog containing ornithine closely parallels the relative substrate activity of its glutamate-containing counterpart. The substitution of ornithine apparently does not perturb the pterin specificity of FPGS. The close parallel between substrate and inhibitor specificity may thus allow the use of currently available structure-activity studies on FPGS to design more potent and more selective inhibitors of FPGS.
McGuire
J J
JJ
Grace Cancer Drug Center, Roswell Park Memorial Institute, Buffalo, NY 14263.
Bolanowska
W E
WE
Piper
J R
JR
D042931:Aldehyde Oxidase; D000445:Aldehyde Oxidoreductases / Q000502:physiology; D000818:Animals; D005492:Folic Acid / Q000031:analogs & derivatives*; D006801:Humans; D007942:Leukemia, Experimental / Q000473:pathology; D008727:Methotrexate / Q000378:metabolism; D009952:Ornithine / Q000031:analogs & derivatives*; D010453:Peptide Synthases / Q000037:antagonists & inhibitors*; D011817:Rabbits; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D005492:Folic Acid; D009952:Ornithine; D000445:Aldehyde Oxidoreductases; D042931:Aldehyde Oxidase; D010453:Peptide Synthases; C023103:folylpolyglutamate synthetase; D008727:Methotrexate