Abstract
6-Substituted classical pyrrolo[2,3-d]pyrimidine antifolates with a three- to six-carbon bridge between the heterocycle and the benzoyl-L-glutamate (compounds 2-5, respectively) were synthesized starting from methyl 4-formylbenzoate and a Wittig reaction with the appropriate triphenylphosphonium bromide, followed by reduction and conversion to the alpha-bromomethylketones. Cyclocondensation of 2,4-diamino-4-oxopyrimidine with the alpha-bromoketones, coupling with diethyl-L-glutamate, and saponification afforded 2-5. Compounds 2-5 had negligible substrate activity for RFC but showed variably potent (nanomolar) and selective inhibitory activities toward Chinese hamster ovary cells that expressed FRalpha or FRbeta and toward FRalpha-expressing KB and IGROV1 human tumor cells. Inhibition of KB cell colony formation was also observed. Glycinamide ribonucleotide formyl transferase (GARFTase) was identified as the primary intracellular target of the pyrrolo[2,3-d]pyrimidines. The combined properties of selective FR targeting, lack of RFC transport, and GARFTase inhibition resulting in potent antitumor activity are unprecedented and warrant development of these analogues as antitumor agents.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Animals
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Antineoplastic Agents / chemical synthesis*
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Antineoplastic Agents / chemistry
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Antineoplastic Agents / pharmacology
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CHO Cells
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Carrier Proteins / antagonists & inhibitors
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Cell Line
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Cell Proliferation / drug effects
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Cricetinae
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Cricetulus
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Enzyme Inhibitors / chemical synthesis*
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Enzyme Inhibitors / chemistry
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Enzyme Inhibitors / pharmacology
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Folate Receptors, GPI-Anchored
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Folic Acid Antagonists / chemical synthesis*
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Folic Acid Antagonists / chemistry
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Folic Acid Antagonists / pharmacology
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Humans
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KB Cells
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Membrane Transport Proteins / drug effects
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Phosphoribosylglycinamide Formyltransferase / antagonists & inhibitors*
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Pyrimidines / chemical synthesis*
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Pyrimidines / chemistry
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Pyrimidines / pharmacology
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Pyrroles / chemical synthesis*
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Pyrroles / chemistry
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Pyrroles / pharmacology
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Receptors, Cell Surface / antagonists & inhibitors
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Reduced Folate Carrier Protein
Substances
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Antineoplastic Agents
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Carrier Proteins
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Enzyme Inhibitors
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Folate Receptors, GPI-Anchored
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Folic Acid Antagonists
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Membrane Transport Proteins
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Pyrimidines
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Pyrroles
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Pyrrolo(2,3-d)pyrimidine
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Receptors, Cell Surface
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Reduced Folate Carrier Protein
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SLC19A1 protein, human
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Phosphoribosylglycinamide Formyltransferase