Methotrexate analogues. 32. Chain extension, alpha-carboxyl deletion, and gamma-carboxyl replacement by sulfonate and phosphonate: effect on enzyme binding and cell-growth inhibition.
Analogues of methotrexate (MTX) and aminopterin (AMT) with aminophosphonoalkanoic, aminoalkanesulfonic, and aminoalkanephosphonic acid side chains in place of glutamate were synthesized and tested as inhibitors of folylpolyglutamate synthetase (FPGS) from mouse liver. The aminophosphonoalkanoic acid analogues were also tested as inhibitors of dihydrofolate reductase (DHFR) from L1210 murine leukemia cells and as inhibitors of the growth of MTX-sensitive (L1210) and MTX-resistant (L1210/R81) cells in culture. The optimal number of CH2 groups in aminophosphonoalkanoic acid analogues of AMT was found to be two for both enzyme inhibition and cell growth inhibition but was especially critical for activity against FPGS. Deletion of the alpha-carboxyl also led to diminished anti-FPGS activity in comparison with previously studied homocysteic acid and 2-amino-4-phosphonobutyric acid analogues. In the aminoalkanesulfonic acid analogues of MTX without an alpha-carboxyl, anti-FPGS activity was low and showed minimal variation as the number of CH2 groups between the carboxamide and sulfonate moieties was changed from one to four. In similar aminoalkanephosphonic acid analogues of MTX, anti-FPGS activity was also low, was comparable for two and three CH2 groups between the carboxamide and phosphonate moieties, and was diminished by monoesterification of the phosphonate group. These effects demonstrate that the alpha-carboxyl group of folate analogues is involved in binding to the active site of FPGS, and that an alpha-carboxyl group should be retained as part of the structure of FPGS inhibitors.
Rosowsky
A
A
Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115.
Forsch
R A
RA
Moran
R G
RG
Kohler
W
W
Freisheim
J H
JH
D000630:Aminopterin / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D055598:Chemical Phenomena; D002621:Chemistry; D004351:Drug Resistance; D005493:Folic Acid Antagonists; D007939:Leukemia L1210 / Q000201:enzymology / Q000473:pathology; D008099:Liver / Q000201:enzymology; D008727:Methotrexate / Q000031:analogs & derivatives* / Q000494:pharmacology; D051379:Mice; D063065:Organophosphonates / Q000138:chemical synthesis / Q000494:pharmacology; D010453:Peptide Synthases / Q000037:antagonists & inhibitors*; D013329:Structure-Activity Relationship; D013451:Sulfonic Acids / Q000138:chemical synthesis / Q000494:pharmacology
D005493:Folic Acid Antagonists; D063065:Organophosphonates; D013451:Sulfonic Acids; D010453:Peptide Synthases; C023103:folylpolyglutamate synthetase; D000630:Aminopterin; D008727:Methotrexate