Abstract
The detailed molecular mechanism of action of second-generation BCR-ABL tyrosine kinase inhibitors, including perturbed targets and pathways, should contribute to rationalized therapy in chronic myeloid leukemia (CML) or in other affected diseases. Here, we characterized the target profile of the dual SRC/ABL inhibitor bosutinib employing a two-tiered approach using chemical proteomics to identify natural binders in whole cell lysates of primary CML and K562 cells in parallel to in vitro kinase assays against a large recombinant kinase panel. The combined strategy resulted in a global survey of bosutinib targets comprised of over 45 novel tyrosine and serine/threonine kinases. We have found clear differences in the target patterns of bosutinib in primary CML cells versus the K562 cell line. A comparison of bosutinib with dasatinib across the whole kinase panel revealed overlapping, but distinct, inhibition profiles. Common among those were the SRC, ABL and TEC family kinases. Bosutinib did not inhibit KIT or platelet-derived growth factor receptor, but prominently targeted the apoptosis-linked STE20 kinases. Although in vivo bosutinib is inactive against ABL T315I, we found this clinically important mutant to be enzymatically inhibited in the mid-nanomolar range. Finally, bosutinib is the first kinase inhibitor shown to target CAMK2G, recently implicated in myeloid leukemia cell proliferation.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Aniline Compounds / chemistry
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Aniline Compounds / pharmacology*
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Antineoplastic Agents / pharmacology*
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Calcium-Calmodulin-Dependent Protein Kinase Type 2 / antagonists & inhibitors
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Dasatinib
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Drug Delivery Systems
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Drug Screening Assays, Antitumor
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Fusion Proteins, bcr-abl / antagonists & inhibitors
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Gene Expression Profiling
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Humans
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K562 Cells / drug effects*
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K562 Cells / enzymology
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Leukemia, Myeloid, Accelerated Phase / enzymology*
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Leukemia, Myeloid, Accelerated Phase / pathology
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Leukocytes, Mononuclear / drug effects
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Leukocytes, Mononuclear / enzymology
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Mitogen-Activated Protein Kinases / antagonists & inhibitors
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Neoplasm Proteins / antagonists & inhibitors*
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Nerve Tissue Proteins / antagonists & inhibitors
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Nitriles / chemistry
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Nitriles / pharmacology*
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Protein Kinase Inhibitors / pharmacology*
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Protein Serine-Threonine Kinases / antagonists & inhibitors
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Protein-Tyrosine Kinases / antagonists & inhibitors*
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Proto-Oncogene Proteins c-abl / antagonists & inhibitors
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Pyrimidines / pharmacology
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Quinolines / chemistry
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Quinolines / pharmacology*
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Signal Transduction / drug effects
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Substrate Specificity
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Thiazoles / pharmacology
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src-Family Kinases / antagonists & inhibitors
Substances
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Aniline Compounds
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Antineoplastic Agents
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Neoplasm Proteins
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Nerve Tissue Proteins
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Nitriles
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Protein Kinase Inhibitors
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Pyrimidines
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Quinolines
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Thiazoles
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bosutinib
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STK24 protein, human
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Tec protein-tyrosine kinase
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Protein-Tyrosine Kinases
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Fusion Proteins, bcr-abl
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Proto-Oncogene Proteins c-abl
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src-Family Kinases
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Protein Serine-Threonine Kinases
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CAMK2G protein, human
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Calcium-Calmodulin-Dependent Protein Kinase Type 2
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Mitogen-Activated Protein Kinases
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Dasatinib