Abstract
Selective protein kinase inhibitors were developed on the basis of the unexpected binding mode of 2,6,9-trisubstituted purines to the adenosine triphosphate-binding site of the human cyclin-dependent kinase 2 (CDK2). By iterating chemical library synthesis and biological screening, potent inhibitors of the human CDK2-cyclin A kinase complex and of Saccharomyces cerevisiae Cdc28p were identified. The structural basis for the binding affinity and selectivity was determined by analysis of a three-dimensional crystal structure of a CDK2-inhibitor complex. The cellular effects of these compounds were characterized in mammalian cells and yeast. In the latter case the effects were characterized on a genome-wide scale by monitoring changes in messenger RNA levels in treated cells with high-density oligonucleotide probe arrays. Purine libraries could provide useful tools for analyzing a variety of signaling and regulatory pathways and may lead to the development of new therapeutics.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Adenine / analogs & derivatives*
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Adenine / chemistry
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Adenine / metabolism
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Adenine / pharmacology
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Binding Sites
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CDC2-CDC28 Kinases*
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CDC28 Protein Kinase, S cerevisiae / antagonists & inhibitors
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Cell Division / drug effects
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Crystallography, X-Ray
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Cyclin A / metabolism
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Cyclin-Dependent Kinase 2
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Cyclin-Dependent Kinases / antagonists & inhibitors*
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Drug Evaluation, Preclinical
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Flavonoids / chemistry
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Flavonoids / metabolism
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Flavonoids / pharmacology
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Gene Expression Regulation, Fungal / drug effects
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Genes, Fungal
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Humans
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Hydrogen Bonding
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Oligonucleotide Probes
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Phosphates / metabolism
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Piperidines / chemistry
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Piperidines / metabolism
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Piperidines / pharmacology
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Protein Serine-Threonine Kinases / antagonists & inhibitors
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Purines / chemical synthesis
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Purines / chemistry
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Purines / metabolism
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Purines / pharmacology*
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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Saccharomyces cerevisiae / enzymology
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Saccharomyces cerevisiae / genetics
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Structure-Activity Relationship
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Transcription, Genetic / drug effects
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Tumor Cells, Cultured
Substances
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Cyclin A
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Flavonoids
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Oligonucleotide Probes
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Phosphates
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Piperidines
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Purines
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RNA, Messenger
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purvalanol B
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alvocidib
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Protein Serine-Threonine Kinases
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CDC2-CDC28 Kinases
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CDC28 Protein Kinase, S cerevisiae
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CDK2 protein, human
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Cyclin-Dependent Kinase 2
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Cyclin-Dependent Kinases
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Adenine