title: The effect of side-chain, para-aminobenzoyl region, and B-ring modifications on dihydrofolate reductase binding, influx via the reduced folate carrier, and cytotoxicity of the potent nonpolyglutamatable antifolate N(alpha)-(4-amino-4-deoxypteroyl)-N(delta)-hemiphthaloyl-L- ornithine.
abstract: N(alpha)-(4-Amino-4-deoxypteroyl)-N(delta)-hemiphthaloyl-L-o rnithine (PT523) is an unusually tight-binding dihydrofolate reductase (DHFR) inhibitor and is efficiently taken up into cells via the reduced folate carrier (RFC). Unlike classical DHFR inhibitors with a glutamate side chain, such as methotrexate and aminopterin, PT523 cannot form polyglutamates. Thus, it resembles lipophilic antifolates such as trimetrexate in not requiring metabolic activation by folylpolyglutamate synthetase in order to produce its antifolate effect. However, in contrast to trimetrexate, PT523 retains growth inhibitory activity in cells with the multidrug resistance phenotype. As part of the preclinical development of this drug, we have performed systematic modification of several regions of the PT523 molecule, with the aim of defining the optimal structural features for DHFR binding, influx into cells via the RFC, and the ability to inhibit cell growth. The following structure-activity correlations have emerged from this ongoing investigation, and are discussed: (1) the hemiphthaloylornithine side chain has the optimal length; (2) the preferred location of the aromatic carboxyl group is the ortho position; and (3) replacement of the phenyl ring of the para-aminobenzoic acid moiety by naphthalene, of nitrogen at the 10-position of the bridge by carbon, and of nitrogen at the 5- and/or 8-position of the B-ring by carbon are all well tolerated. Several of the second generation analogs of PT523 are more potent DHFR inhibitors and better RFC substrates than PT523 itself, and are more potent inhibitors of tumor cell growth in culture.
authors.affiliation: Dana-Farber Cancer Institute and Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 44 Binney Street, Boston, MA, USA. andre_rosowsky@dfci.harvard.edu
authors.forename: A
authors.initials: A
authors.lastname: Rosowsky
authors.forename: J E
authors.initials: JE
authors.lastname: Wright
authors.forename: C M
authors.initials: CM
authors.lastname: Vaidya
authors.forename: R A
authors.initials: RA
authors.lastname: Forsch
mesh_terms: D010129:4-Aminobenzoic Acid / Q000737:chemistry / Q000378:metabolism; D000630:Aminopterin / Q000031:analogs & derivatives / Q000737:chemistry; D000970:Antineoplastic Agents / Q000493:pharmacokinetics* / Q000494:pharmacology; D001665:Binding Sites; D002455:Cell Division; D018432:Drug Resistance, Multiple; D004354:Drug Screening Assays, Antitumor; D005492:Folic Acid / Q000378:metabolism; D005493:Folic Acid Antagonists / Q000493:pharmacokinetics* / Q000494:pharmacology; D006801:Humans; D009952:Ornithine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000494:pharmacology; D011622:Pterins / Q000493:pharmacokinetics* / Q000494:pharmacology; D019163:Reducing Agents; D013329:Structure-Activity Relationship; D013762:Tetrahydrofolate Dehydrogenase / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D005493:Folic Acid Antagonists; D011622:Pterins; D019163:Reducing Agents; C085590:N(alpha)-(4-amino-4-deoxypteroyl)-N(delta)-hemiphthaloyl-L-ornithine; D005492:Folic Acid; D009952:Ornithine; D013762:Tetrahydrofolate Dehydrogenase; D000630:Aminopterin; D010129:4-Aminobenzoic Acid