Abstract

The proposed project is part of a long range research effort aimed at the development of new approaches to the chemotherapy of neoplastic diseases. The main objective of this research is to design novel target-oriented antifolates based on biochemical and mechanistic rationales derived from structural and functional information and to study their effects at the cellular and molecular level. As specific targets, the proposed study focuses on enzymes of two interrelated metabolic cycles, one responsible for the formation and breakdown of the poly-gamma-glutamates of folates and antifolates and the other the folylpolyglutamate dependent biosynthesis of thymidylate (the TS cycle). The proposed research sets the following specific aims: To design and synthesize potential inhibitors of folylpolyglutamate synthetase, gamma-glutamyl hydrolase (conjugase), thymidylate synthase, dihydrofolate reductase and serine hydroxymethyl transferase; to evaluate the effects of the inhibitors on their respective targets in cellular enzyme systems; to determine the growth inhibitory effects of the target compounds on L1210 mouse and CCRF-CEM human leukemia cells in culture; to study the mechanism of selected inhibitor-enzyme interactions at the molecular level using techniques of enzymology, X-ray crystallography, molecular mechanics and computer graphics; to study the polyglutamylation of active antifolates and its importance for biological activity; to correlate cellular and cell-free enzyme inhibitory activity with in vitro cytotoxicity data and based on these correlations select candidates for in vivo biological testing. The study of drug-enzyme interactions may further our knowledge of the mechanisms of folate dependent enzymes and furnish new rationales for inhibitor design. Since folate metabolism is intimately linked to nucleic acid biosynthesis and cellular proliferation, this research may lead to important observations in tumor biology and new approaches to cancer treatment.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA035212-09
Application #
3172825
Study Section
Experimental Therapeutics Subcommittee 1 (ET)
Project Start
1983-09-30
Project End
1997-03-31
Budget Start
1993-04-01
Budget End
1994-03-31
Support Year
9
Fiscal Year
1993
Total Cost
Indirect Cost

  Institution

Name
State University of New York at Buffalo
Department
Type
Schools of Pharmacy
DUNS #
038633251
City
Buffalo
State
NY
Country
United States
Zip Code
14260

  Publications

Saxl, Ruth L; Reston, James; Nie, Zhe et al. (2003) Modification of Escherichia coli thymidylate synthase at tyrosine-94 by 5-imidazolylpropynyl-2'-deoxyuridine 5'-monophosphate. Biochemistry 42:4544-51
Maurer, T S; Pan, J; Booth, B P et al. (2000) Examination of N-hydroxylation as a prerequisite mechanism of nitric oxide synthase inactivation. Bioorg Med Chem Lett 10:1077-80
Mao, Z; Pan, J; Kalman, T I (1996) Design and synthesis of histidine analogues of folic acid and methotrexate as potential folylpolyglutamate synthetase inhibitors. J Med Chem 39:4340-4
Abraham, T W; Kalman, T I; McIntee, E J et al. (1996) Synthesis and biological activity of aromatic amino acid phosphoramidates of 5-fluoro-2'-deoxyuridine and 1-beta-arabinofuranosylcytosine: evidence of phosphoramidase activity. J Med Chem 39:4569-75
Kalman, T I (1993) Mechanism-based approaches to inhibition of the synthesis and degradation of folate and antifolate polyglutamates. Adv Exp Med Biol 338:639-43
Cody, V; Luft, J R; Ciszak, E et al. (1992) Crystal structure determination at 2.3 A of recombinant human dihydrofolate reductase ternary complex with NADPH and methotrexate-gamma-tetrazole. Anticancer Drug Des 7:483-91
Wagh, P V; Kalman, T I (1992) A rapid colorimetric assay for gamma-glutamyl hydrolase (conjugase). Anal Biochem 207:1-5
Harvison, P J; Kalman, T I (1992) Synthesis and biological activity of novel folic acid analogues: pteroyl-S-alkylhomocysteine sulfoximines. J Med Chem 35:1227-33
Kalman, T I; Marinelli, E R; Xu, B et al. (1991) Inhibition of cellular thymidylate synthesis by cytotoxic propenal derivatives of pyrimidine bases and deoxynucleosides. Biochem Pharmacol 42:431-7
McGuire, J J; Russell, C A; Bolanowska, W E et al. (1990) Biochemical and growth inhibition studies of methotrexate and aminopterin analogues containing a tetrazole ring in place of the gamma-carboxyl group. Cancer Res 50:1726-31

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