title: Exploitation of folate and antifolate polyglutamylation to achieve selective anticancer chemotherapy.
abstract: Synthesis of poly(gamma-glutamate) metabolites of natural folates and antifolates is a critical process. Folypolyglutamates are essential for cell proliferation. Polyglutamates of glutamate (Glu)-containing antifolates are often critical for their cytotoxic action and are relevant to antifolate resistance. However, the role of polyglutamate synthesis in selectivity is less clear. We have undertaken a research program to further define the significance of polyglutamate metabolism and to devise ways to exploit this metabolism to achieve greater therapeutic selectivity in cancer chemotherapy. This article briefly reviews several approaches tested thus far. Inhibition of folypolyglutamate synthesis should lead to cell death. Current ornithine (Orn)-containing folate-based inhibitors of the enzyme responsible for their synthesis, folypolyglutamate synthetase (FPGS), are poorly transported, apparently because of interference by the protonated delta-amine. Replacement of Orn with 4, 4-difluoroOrn, the delta-amine of which has a much lower pKa and is thus less protonated at physiological pH, was explored. Since it is unclear how polyglutamylation contributes to selectivity, we explored generic means either to eliminate or to enhance polyglutamylation. The data indicate that substitution for Glu in an antifolate by some Glu analogs in which the gamma-COOH is either altered or replaced (e.g., gamma-tetrazole-Glu) leads to loss of both FPGS substrate activity and binding; antifolate target specificity is unchanged, while uptake is actually enhanced. Substitution of 3,3-difluoroGlu for Glu leads to enhanced polyglutamylation (although probably only to the diglutamate), retention of target specificity, and at least equal uptake. Comparative studies of the same antifolate containing different replacements for Glu, such as gamma-tetrazole-Glu (no polyglutamylation) or 3,3-difluoroGlu (enhanced polyglutamylation), will be useful in exploring the role and significance of polyglutamylation.
authors.affiliation: Grace Cancer Drug Center, Roswell Park Cancer Institute, Buffalo, NY, USA.
authors.forename: J J
authors.initials: JJ
authors.lastname: McGuire
authors.forename: T
authors.initials: T
authors.lastname: Tsukamoto
authors.forename: B P
authors.initials: BP
authors.lastname: Hart
authors.forename: J K
authors.initials: JK
authors.lastname: Coward
authors.forename: T I
authors.initials: TI
authors.lastname: Kalman
authors.forename: J
authors.initials: J
authors.lastname: Galivan
mesh_terms: D000970:Antineoplastic Agents / Q000627:therapeutic use*; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000627:therapeutic use*; D005493:Folic Acid Antagonists / Q000378:metabolism / Q000627:therapeutic use*; D006801:Humans; D015394:Molecular Structure; D010453:Peptide Synthases / Q000037:antagonists & inhibitors*; D011624:Pteroylpolyglutamic Acids / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D005493:Folic Acid Antagonists; D011624:Pteroylpolyglutamic Acids; D010453:Peptide Synthases; C023103:folylpolyglutamate synthetase