Global target profile of the kinase inhibitor bosutinib in primary chronic myeloid leukemia cells.
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.
Remsing Rix
L L
LL
Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
Rix
U
U
Colinge
J
J
Hantschel
O
O
Bennett
K L
KL
Stranzl
T
T
Müller
A
A
Baumgartner
C
C
Valent
P
P
Augustin
M
M
Till
J H
JH
Superti-Furga
G
G
D000814:Aniline Compounds / Q000737:chemistry / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000037:antagonists & inhibitors; D000069439:Dasatinib; D016503:Drug Delivery Systems; D004354:Drug Screening Assays, Antitumor; D016044:Fusion Proteins, bcr-abl / Q000037:antagonists & inhibitors; D020869:Gene Expression Profiling; D006801:Humans; D020014:K562 Cells / Q000187:drug effects* / Q000201:enzymology; D015465:Leukemia, Myeloid, Accelerated Phase / Q000201:enzymology* / Q000473:pathology; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000201:enzymology; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors*; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors; D009570:Nitriles / Q000737:chemistry / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D016315:Proto-Oncogene Proteins c-abl / Q000037:antagonists & inhibitors; D011743:Pyrimidines / Q000494:pharmacology; D011804:Quinolines / Q000737:chemistry / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D013379:Substrate Specificity; D013844:Thiazoles / Q000494:pharmacology; D019061:src-Family Kinases / Q000037:antagonists & inhibitors; D000099317:Hippo Kinases
D000814:Aniline Compounds; D000970:Antineoplastic Agents; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D000069439:Dasatinib; D016044:Fusion Proteins, bcr-abl; D020928:Mitogen-Activated Protein Kinases; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D009570:Nitriles; D047428:Protein Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases; D016315:Proto-Oncogene Proteins c-abl; D011743:Pyrimidines; D011804:Quinolines; D013844:Thiazoles; D019061:src-Family Kinases; C471992:bosutinib; C109163:STK24 protein, human; D000099317:Hippo Kinases; C089848:Tec protein-tyrosine kinase; C517662:CAMK2G protein, human