Dual-specific Src and Abl kinase inhibitors, PP1 and CGP76030, inhibit growth and survival of cells expressing imatinib mesylate-resistant Bcr-Abl kinases.
The leukemogenic tyrosine kinase Bcr-Abl contains a highly conserved inhibitor-binding pocket (IBP), which serves as a binding site for imatinib mesylate. Mutations at the IBP may lead to resistance of the Abl kinase against imatinib mesylate. To examine the mechanisms of imatinib mesylate binding and resistance in more detail, we created several point mutations at amino acid positions 315 and 380 of Abl, blocking the access to the IBP and rendering Bcr-Abl imatinib mesylate-resistant. Moreover, introduction of a mutation destabilizing the inactive conformation of Abl (Asp276Ser/Glu279Ser) also led to imatinib mesylate resistance, suggesting that the inhibitor required inactivation of the kinase prior to binding. These Bcr-Abl mutants were then used to evaluate the binding mode and specificity of 2 compounds, PP1 and CGP76030, originally characterized as Src kinase inhibitors. Both compounds inhibited Bcr-Abl in a concentration-dependent manner by overlapping binding modes. However, in contrast to imatinib mesylate, PP1 and CGP76030 blocked cell growth and survival in cells expressing various inhibitor-resistant Abl mutants. Studies on the potential signaling mechanisms demonstrated that in cells expressing inhibitor-resistant Bcr-Abl mutants, PP1 and CGP76030 inhibited the activity of Src family tyrosine kinases and Akt but not signal transducer and activator of transcription-5 (STAT5) and JUN kinase (Jnk). The results suggest that the use of Src kinase inhibitors is a potential strategy to prevent or overcome clonal evolution of imatinib mesylate resistance in Bcr-Abl(+) leukemia.
Warmuth
Markus
M
Klinische Kooperationsgruppe für Gentherapie, GSF-Forschungszentrum für Umwelt und Gesundheit, Munich, Germany.
Simon
Nicola
N
Mitina
Olga
O
Mathes
Ruth
R
Fabbro
Doriano
D
Manley
Paul W
PW
Buchdunger
Elisabeth
E
Forster
Karin
K
Moarefi
Ismail
I
Hallek
Michael
M
D017209:Apoptosis / Q000187:drug effects; D001549:Benzamides; D001665:Binding Sites / Q000235:genetics; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D016044:Fusion Proteins, bcr-abl / Q000737:chemistry / Q000235:genetics; D000068877:Imatinib Mesylate; D007938:Leukemia / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D016374:Oncogene Proteins v-abl / Q000037:antagonists & inhibitors*; D010879:Piperazines; D011487:Protein Conformation; D011720:Pyrazoles / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*; D011758:Pyrroles / Q000494:pharmacology; D019061:src-Family Kinases / Q000037:antagonists & inhibitors*
C108123:4-amino-5-(4-methylphenyl)-7-(tert-butyl)pyrazolo(3,4-d)pyrimidine; D001549:Benzamides; C472714:CGP 76030; D004791:Enzyme Inhibitors; D016374:Oncogene Proteins v-abl; D010879:Piperazines; D011720:Pyrazoles; D011743:Pyrimidines; D011758:Pyrroles; D000068877:Imatinib Mesylate; D016044:Fusion Proteins, bcr-abl; D019061:src-Family Kinases