Exploiting chemical libraries, structure, and genomics in the search for kinase inhibitors.
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.
Howard Hughes Medical Institute, University of California, Berkeley, CA 94720, USA.
N S
NS
Gray
L
L
Wodicka
A M
AM
Thunnissen
T C
TC
Norman
S
S
Kwon
F H
FH
Espinoza
D O
DO
Morgan
G
G
Barnes
S
S
LeClerc
L
L
Meijer
S H
SH
Kim
D J
DJ
Lockhart
P G
PG
Schultz
D000225:Adenine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D001665:Binding Sites; D042846:CDC2-CDC28 Kinases; D017489:CDC28 Protein Kinase, S cerevisiae / Q000037:antagonists & inhibitors; D002455:Cell Division / Q000187:drug effects; D018360:Crystallography, X-Ray; D019925:Cyclin A / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors*; D004353:Drug Evaluation, Preclinical; D005419:Flavonoids / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005800:Genes, Fungal; D006801:Humans; D006860:Hydrogen Bonding; D015345:Oligonucleotide Probes; D010710:Phosphates / Q000378:metabolism; D010880:Piperidines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors; D011687:Purines / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured
D019925:Cyclin A; D005419:Flavonoids; D015345:Oligonucleotide Probes; D010710:Phosphates; D010880:Piperidines; D011687:Purines; D012333:RNA, Messenger; C113584:purvalanol B; C077990:alvocidib; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; D017489:CDC28 Protein Kinase, S cerevisiae; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases; D000225:Adenine