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1.
Nat Chem Biol ; 10(4): 305-12, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24584101

ABSTRACT

Concomitant inhibition of multiple cancer-driving kinases is an established strategy to improve the durability of clinical responses to targeted therapies. The difficulty of discovering kinase inhibitors with an appropriate multitarget profile has, however, necessitated the application of combination therapies, which can pose major clinical development challenges. Epigenetic reader domains of the bromodomain family have recently emerged as new targets for cancer therapy. Here we report that several clinical kinase inhibitors also inhibit bromodomains with therapeutically relevant potencies and are best classified as dual kinase-bromodomain inhibitors. Nanomolar activity on BRD4 by BI-2536 and TG-101348, which are clinical PLK1 and JAK2-FLT3 kinase inhibitors, respectively, is particularly noteworthy as these combinations of activities on independent oncogenic pathways exemplify a new strategy for rational single-agent polypharmacological targeting. Furthermore, structure-activity relationships and co-crystal structures identify design features that enable a general platform for the rational design of dual kinase-bromodomain inhibitors.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/chemical synthesis , Antineoplastic Combined Chemotherapy Protocols/pharmacology , Drug Design , Polypharmacology , Protein Kinase Inhibitors/chemical synthesis , Protein Kinase Inhibitors/pharmacology , Blotting, Western , Calorimetry , Cell Line, Tumor , Crystallization , Drug Interactions , Drug Screening Assays, Antitumor , Epigenesis, Genetic , High-Throughput Screening Assays , Humans , Neoplasms/drug therapy , Neoplasms/genetics , Proto-Oncogene Proteins c-myc/metabolism , Pteridines/pharmacology , Pyrrolidines/pharmacology , Recombinant Proteins/pharmacology , Signal Transduction/drug effects , Structure-Activity Relationship , Sulfonamides/pharmacology
2.
Nat Biotechnol ; 29(11): 1046-51, 2011 Oct 30.
Article in English | MEDLINE | ID: mdl-22037378

ABSTRACT

We tested the interaction of 72 kinase inhibitors with 442 kinases covering >80% of the human catalytic protein kinome. Our data show that, as a class, type II inhibitors are more selective than type I inhibitors, but that there are important exceptions to this trend. The data further illustrate that selective inhibitors have been developed against the majority of kinases targeted by the compounds tested. Analysis of the interaction patterns reveals a class of 'group-selective' inhibitors broadly active against a single subfamily of kinases, but selective outside that subfamily. The data set suggests compounds to use as tools to study kinases for which no dedicated inhibitors exist. It also provides a foundation for further exploring kinase inhibitor biology and toxicity, as well as for studying the structural basis of the observed interaction patterns. Our findings will help to realize the direct enabling potential of genomics for drug development and basic research about cellular signaling.


Subject(s)
Protein Kinase Inhibitors/chemistry , Protein Kinases/chemistry , Catalysis , Drug Design , Enzyme Stability , High-Throughput Screening Assays , Humans , Protein Binding , Protein Kinase Inhibitors/classification , Protein Kinases/classification , Proteomics , Signal Transduction , Substrate Specificity
3.
Chem Biol ; 17(11): 1241-9, 2010 Nov 24.
Article in English | MEDLINE | ID: mdl-21095574

ABSTRACT

Interactions between kinases and small molecule inhibitors can be activation state dependent. A detailed understanding of inhibitor binding therefore requires characterizing interactions across multiple activation states. We have systematically explored the effects of ABL1 activation loop phosphorylation and PDGFR family autoinhibitory juxtamembrane domain docking on inhibitor binding affinity. For a diverse compound set, the affinity patterns correctly classify inhibitors as having type I or type II binding modes, and we show that juxtamembrane domain docking can have dramatic negative effects on inhibitor affinity. The results have allowed us to associate ligand-induced conformational changes observed in cocrystal structures with specific energetic costs. The approach we describe enables investigation of the complex relationship between kinase activation state and compound binding affinity and should facilitate strategic inhibitor design.


Subject(s)
Protein Kinase Inhibitors/chemistry , Proto-Oncogene Proteins c-abl/antagonists & inhibitors , Small Molecule Libraries/chemistry , Amino Acid Sequence , Computer Simulation , Molecular Sequence Data , Phosphorylation , Protein Binding , Protein Kinase Inhibitors/pharmacology , Protein Structure, Tertiary , Proto-Oncogene Proteins c-abl/metabolism , Proto-Oncogene Proteins c-kit/antagonists & inhibitors , Proto-Oncogene Proteins c-kit/metabolism , Receptor, Macrophage Colony-Stimulating Factor/antagonists & inhibitors , Receptor, Macrophage Colony-Stimulating Factor/metabolism , Receptor, Platelet-Derived Growth Factor beta/chemistry , Receptor, Platelet-Derived Growth Factor beta/metabolism , Small Molecule Libraries/pharmacology , fms-Like Tyrosine Kinase 3/antagonists & inhibitors , fms-Like Tyrosine Kinase 3/metabolism
4.
Nat Biotechnol ; 26(1): 127-32, 2008 Jan.
Article in English | MEDLINE | ID: mdl-18183025

ABSTRACT

Kinase inhibitors are a new class of therapeutics with a propensity to inhibit multiple targets. The biological consequences of multi-kinase activity are poorly defined, and an important step toward understanding the relationship between selectivity, efficacy and safety is the exploration of how inhibitors interact with the human kinome. We present interaction maps for 38 kinase inhibitors across a panel of 317 kinases representing >50% of the predicted human protein kinome. The data constitute the most comprehensive study of kinase inhibitor selectivity to date and reveal a wide diversity of interaction patterns. To enable a global analysis of the results, we introduce the concept of a selectivity score as a general tool to quantify and differentiate the observed interaction patterns. We further investigate the impact of panel size and find that small assay panels do not provide a robust measure of selectivity.


Subject(s)
Phosphotransferases/antagonists & inhibitors , Protein Interaction Mapping/methods , Protein Kinase Inhibitors/chemistry , Proteome/chemistry , Quantitative Structure-Activity Relationship , Binding Sites , Enzyme Activation , Humans , Protein Binding
5.
Proc Natl Acad Sci U S A ; 102(31): 11011-6, 2005 Aug 02.
Article in English | MEDLINE | ID: mdl-16046538

ABSTRACT

To realize the full potential of targeted protein kinase inhibitors for the treatment of cancer, it is important to address the emergence of drug resistance in treated patients. Mutant forms of BCR-ABL, KIT, and the EGF receptor (EGFR) have been found that confer resistance to the drugs imatinib, gefitinib, and erlotinib. The mutations weaken or prevent drug binding, and interestingly, one of the most common sites of mutation in all three kinases is a highly conserved "gatekeeper" threonine residue near the kinase active site. We have identified existing clinical compounds that bind and inhibit drug-resistant mutant variants of ABL, KIT, and EGFR. We found that the Aurora kinase inhibitor VX-680 and the p38 inhibitor BIRB-796 inhibit the imatinib- and BMS-354825-resistant ABL(T315I) kinase. The KIT/FLT3 inhibitor SU-11248 potently inhibits the imatinib-resistant KIT(V559D/T670I) kinase, consistent with the clinical efficacy of SU-11248 against imatinib-resistant gastrointestinal tumors, and the EGFR inhibitors EKB-569 and CI-1033, but not GW-572016 and ZD-6474, potently inhibit the gefitinib- and erlotinib-resistant EGFR(L858R/T790M) kinase. EKB-569 and CI-1033 are already in clinical trials, and our results suggest that they should be considered for testing in the treatment of gefitinib/erlotinib-resistant non-small cell lung cancer. The results highlight the strategy of screening existing clinical compounds against newly identified drug-resistant mutant variants to find compounds that may serve as starting points for the development of next-generation drugs, or that could be used directly to treat patients that have acquired resistance to first-generation targeted therapy.


Subject(s)
ErbB Receptors/antagonists & inhibitors , ErbB Receptors/genetics , Oncogene Proteins v-abl/antagonists & inhibitors , Oncogene Proteins v-abl/genetics , Protein Kinase Inhibitors/pharmacology , Proto-Oncogene Proteins c-kit/genetics , Proto-Oncogene Proteins c-kit/metabolism , Aminoquinolines , Aniline Compounds , Cell Line , Drug Resistance, Neoplasm/genetics , Humans , Indoles/pharmacology , Kinetics , Morpholines/pharmacology , Mutation , Naphthalenes/pharmacology , Neoplasms/drug therapy , Neoplasms/enzymology , Neoplasms/genetics , Organic Chemicals/pharmacology , Piperazines/pharmacology , Pyrazoles/pharmacology , Pyrroles/pharmacology , Sunitinib
6.
Proc Natl Acad Sci U S A ; 102(29): 10357-62, 2005 Jul 19.
Article in English | MEDLINE | ID: mdl-16002470

ABSTRACT

The current model to explain the organization of the mammalian nervous system is based on studies of anatomy, embryology, and evolution. To further investigate the molecular organization of the adult mammalian brain, we have built a gene expression-based brain map. We measured gene expression patterns for 24 neural tissues covering the mouse central nervous system and found, surprisingly, that the adult brain bears a transcriptional "imprint" consistent with both embryological origins and classic evolutionary relationships. Embryonic cellular position along the anterior-posterior axis of the neural tube was shown to be closely associated with, and possibly a determinant of, the gene expression patterns in adult structures. We also observed a significant number of embryonic patterning and homeobox genes with region-specific expression in the adult nervous system. The relationships between global expression patterns for different anatomical regions and the nature of the observed region-specific genes suggest that the adult brain retains a degree of overall gene expression established during embryogenesis that is important for regional specificity and the functional relationships between regions in the adult. The complete collection of extensively annotated gene expression data along with data mining and visualization tools have been made available on a publicly accessible web site (www.barlow-lockhart-brainmapnimhgrant.org).


Subject(s)
Biological Evolution , Central Nervous System/metabolism , Gene Expression Profiling , Gene Expression Regulation, Developmental , Mice/metabolism , Models, Neurological , Algorithms , Animals , Cluster Analysis , Databases, Genetic , Mice/embryology , Microarray Analysis
7.
Nat Biotechnol ; 23(3): 329-36, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15711537

ABSTRACT

Kinase inhibitors show great promise as a new class of therapeutics. Here we describe an efficient way to determine kinase inhibitor specificity by measuring binding of small molecules to the ATP site of kinases. We have profiled 20 kinase inhibitors, including 16 that are approved drugs or in clinical development, against a panel of 119 protein kinases. We find that specificity varies widely and is not strongly correlated with chemical structure or the identity of the intended target. Many novel interactions were identified, including tight binding of the p38 inhibitor BIRB-796 to an imatinib-resistant variant of the ABL kinase, and binding of imatinib to the SRC-family kinase LCK. We also show that mutations in the epidermal growth factor receptor (EGFR) found in gefitinib-responsive patients do not affect the binding affinity of gefitinib or erlotinib. Our results represent a systematic small molecule-protein interaction map for clinical compounds across a large number of related proteins.


Subject(s)
Drug Design , Escherichia coli Proteins/metabolism , Escherichia coli/metabolism , Pharmaceutical Preparations/metabolism , Piperazines/metabolism , Protein Interaction Mapping/methods , Protein Kinase Inhibitors/metabolism , Pyrimidines/metabolism , Benzamides , Imatinib Mesylate , Microchemistry/methods , Protein Binding
8.
Proc Natl Acad Sci U S A ; 99(24): 15669-74, 2002 Nov 26.
Article in English | MEDLINE | ID: mdl-12441396

ABSTRACT

We have examined the progression of hepatitis C virus (HCV) infections by gene expression analysis of liver biopsies in acutely infected chimpanzees that developed persistent infection, transient viral clearance, or sustained clearance. Both common responses and outcome-specific changes in expression were observed. All chimpanzees showed gene expression patterns consistent with an IFN-alpha response that correlated with the magnitude and duration of infection. Transient and sustained viral clearance were uniquely associated with induction of IFN-gamma-induced genes and other genes involved in antigen processing and presentation and the adaptive immune response. During the early stages of infection, host genes involved in lipid metabolism were also differentially regulated. We also show that drugs that affect these biosynthetic pathways can regulate HCV replication in HCV replicon systems. Our results reveal genome-wide transcriptional changes that reflect the establishment, spread, and control of infection, and they reveal potentially unique antiviral programs associated with clearance of HCV infection.


Subject(s)
Gene Expression Profiling , Gene Expression Regulation , Hepatitis C/metabolism , Animals , Antigen Presentation , Fatty Acids/metabolism , Genes, Reporter , Hepacivirus/drug effects , Hepacivirus/genetics , Hepacivirus/physiology , Hepatitis C/genetics , Hepatitis C/immunology , Interferons/biosynthesis , Lipid Metabolism , Pan troglodytes , Replicon , Transcription, Genetic , Transfection , Viremia/genetics , Viremia/immunology , Virus Replication/drug effects
9.
Genome Res ; 12(8): 1210-20, 2002 Aug.
Article in English | MEDLINE | ID: mdl-12176929

ABSTRACT

Short open reading frames (ORFs) occur frequently in primary genome sequence. Distinguishing bona fide small genes from the tens of thousands of short ORFs is one of the most challenging aspects of genome annotation. Direct experimental evidence is often required. Here we use a combination of expression profiling and mass spectrometry to verify the independent transcription of 138 and the translation of 50 previously nonannotated genes in the Saccharomyces cerevisiae genome. Through combined evidence, we propose the addition of 62 new genes to the genome and provide experimental support for the inclusion of 10 previously identified genes.


Subject(s)
Genes, Fungal/genetics , Genome, Fungal , Saccharomyces cerevisiae/genetics , Amino Acid Sequence , Animals , Chromosome Mapping , Chromosomes, Fungal/genetics , Computational Biology/methods , Conserved Sequence/genetics , Conserved Sequence/physiology , Databases, Genetic/classification , Drosophila Proteins/genetics , Gene Expression Profiling/methods , Genes, Fungal/physiology , Mice , Molecular Sequence Data , Open Reading Frames/genetics , Open Reading Frames/physiology , Peptides/analysis , Peptides/physiology , Protein Biosynthesis/genetics , Protein Biosynthesis/physiology , Saccharomyces cerevisiae Proteins/analysis , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae Proteins/physiology , Sequence Alignment , Sequence Homology, Amino Acid , Transcription, Genetic/physiology
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