title: 10-Formyl-5,10-dideaza-acyclic-5,6,7,8-tetrahydrofolic acid (10-formyl-DDACTHF): a potent cytotoxic agent acting by selective inhibition of human GAR Tfase and the de novo purine biosynthetic pathway.
abstract: The synthesis of 10-formyl-DDACTHF (3) as a potential inhibitor of glycinamide ribonucleotide transformylase (GAR Tfase) and aminoimidazole carboxamide ribonucleotide transformylase (AICAR Tfase) is reported. Aldehyde 3, the corresponding gamma- and alpha-pentaglutamates 21 and 25 and related agents were evaluated for inhibition of folate-dependent enzymes including GAR Tfase and AICAR Tfase. The inhibitors were found to exhibit potent cytotoxic activity (CCRF-CEM IC(50) for 3=60nM) that exceeded their enzyme inhibition potency [K(i) (3)=6 and 1 microM for Escherichia coli GAR and human AICAR Tfase, respectively]. Cytotoxicity rescue by medium purines, but not pyrimidines, indicated that the potent cytotoxic activity is derived from selective purine biosynthesis inhibition and rescue by AICAR monophosphate established that the activity is derived preferentially from GAR versus AICAR Tfase inhibition. The potent cytotoxic compounds including aldehyde 3 lost activity against CCRF-CEM cell lines deficient in the reduced folate carrier (CCRF-CEM/MTX) or folylpolyglutamate synthase (CCRF-CEM/FPGS(-)) establishing that their potent activity requires both reduced folate carrier transport and polyglutamation. Unexpectedly, the pentaglutamates displayed surprisingly similar K(i)'s versus E. coli GAR Tfase and only modestly enhanced K(i)'s versus human AICAR Tfase. On the surface this initially suggested that the potent cytotoxic activity of 3 and related compounds might be due simply to preferential intracellular accumulation of the inhibitors derived from effective transport and polyglutamation (i.e., ca. 100-fold higher intracellular concentrations). However, a subsequent examination of the inhibitors against recombinant human GAR Tfase revealed they and the corresponding gamma-pentaglutamates were unexpectedly much more potent against the human versus E. coli enzyme (K(i) for 3, 14nM against rhGAR Tfase versus 6 microM against E. coli GAR Tfase) which also accounts for their exceptional cytotoxic potency.
authors.affiliation: Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Marsilje
authors.forename: Marc A
authors.initials: MA
authors.lastname: Labroli
authors.forename: Michael P
authors.initials: MP
authors.lastname: Hedrick
authors.forename: Qing
authors.initials: Q
authors.lastname: Jin
authors.forename: Joel
authors.initials: J
authors.lastname: Desharnais
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Baker
authors.forename: Lata T
authors.initials: LT
authors.lastname: Gooljarsingh
authors.forename: Joseph
authors.initials: J
authors.lastname: Ramcharan
authors.forename: Ali
authors.initials: A
authors.lastname: Tavassoli
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Ian A
authors.initials: IA
authors.lastname: Wilson
authors.forename: G Peter
authors.initials: GP
authors.lastname: Beardsley
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Benkovic
authors.forename: Dale L
authors.initials: DL
authors.lastname: Boger
mesh_terms: D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000494:pharmacology; D002352:Carrier Proteins / Q000502:physiology; D002455:Cell Division / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology; D058973:Folate Receptors, GPI-Anchored; D006801:Humans; D019877:Hydroxymethyl and Formyl Transferases / Q000037:antagonists & inhibitors*; D010453:Peptide Synthases / Q000502:physiology; D051027:Phosphoribosylaminoimidazolecarboxamide Formyltransferase; D051026:Phosphoribosylglycinamide Formyltransferase; D011687:Purines / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D011956:Receptors, Cell Surface; D013329:Structure-Activity Relationship; D013763:Tetrahydrofolates / Q000138:chemical synthesis*; D014407:Tumor Cells, Cultured
chemical_list: C475811:10-formyl-5,10-dideaza-acyclic-5,6,7,8-tetrahydrofolic acid; D000970:Antineoplastic Agents; D002352:Carrier Proteins; D004791:Enzyme Inhibitors; D058973:Folate Receptors, GPI-Anchored; D011687:Purines; D011956:Receptors, Cell Surface; D013763:Tetrahydrofolates; D019877:Hydroxymethyl and Formyl Transferases; D051026:Phosphoribosylglycinamide Formyltransferase; D051027:Phosphoribosylaminoimidazolecarboxamide Formyltransferase; D010453:Peptide Synthases; C023103:folylpolyglutamate synthetase