Effects of AMN107, a Novel Aminopyrimidine Tyrosine Kinase Inhibitor, on Human Mast Cells Bearing Wild-Type or Mutated Codon 816 c-kit.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3528-3528 ◽  
Author(s):  
Srdan Verstovsek ◽  
Cem Akin ◽  
Giles J. Francis ◽  
Manshouri Taghi ◽  
Ly Huynh ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Tyrosine Kinase ◽  
Tyrosine Kinase Inhibitor ◽  
Cell Line ◽  
Cell Lines ◽  
Cellular Proliferation ◽  
Kinase Inhibitor ◽  
Wild Type ◽  
Kit Mutation

Abstract Background. Majority of adult patients with systemic mastocytosis (SM) have activating mutation in codon 816 of c-kit (CD117), a receptor on the surface of mast cells. This abnormality is responsible for the pathogenesis of the disease. Methods. We investigated the effects of a newly designed tyrosine kinase inhibitor, AMN107, by comparing its in vitro inhibitory potency on c-kit mutated mast cell lines and patient samples with that of imatinib mesylate, another tyrosine kinase inhibitor, effective in some patients with SM. Two cell lines, subclones of HMC-1 cells, were used: HMC-1560 carrying juxtamembrane domain mutation in codon 560 of c-kit, and HMC-1560, 816 carrying both codon 560 mutation and tyrosine kinase domain mutation in codon 816 of c-kit. Results. In HMC-1560 mast cell line carrying wild-type codon 816, AMN107 was as potent as imatinib in inhibiting cellular proliferation, with IC50 values of 108 and 74 nM respectively, while in HMC-1560, 816 cell line carrying 816 mutation, neither medication had an effect. AMN107 was also as effective as imatinib in inhibiting phosphorylation of c-kit tyrosine kinase in HMC-1560 cells. The inhibition of cellular proliferation was associated with induction of apoptosis in HMC-1560 cells. AMN107 in concentrations up to 1 uM had no effect on bone marrow mast cells carrying D816V c-kit mutation obtained from patients with mastocytosis. Conclusions. Our results suggest similar potency of AMN107 and imatinib in mast cells that carry wild-type codon 816, but no activity against codon 816 mutation carrying cells.

Tyrosine Kinase Inhibitor STI571 (Imatinib) Cooperates with Wild-Type p53 on K562 Cell Line to Enhance Its Proapoptotic Effects

2005 ◽  
Vol 114 (3) ◽  
pp. 150-154 ◽  
Author(s):  
Gianluca Brusa ◽  
Manuela Mancini ◽  
Fabio Campanini ◽  
Alberto Calabrò ◽  
Elisa Zuffa ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Tyrosine Kinase Inhibitor ◽  
Cell Line ◽  
K562 Cell ◽  
Kinase Inhibitor ◽  
Wild Type ◽  
K562 Cell Line ◽  
Wild Type P53

Effects of AMN107, a novel aminopyrimidine tyrosine kinase inhibitor, on human mast cells bearing wild-type or mutated codon 816 c-kit

2006 ◽  
Vol 30 (11) ◽  
pp. 1365-1370 ◽  
Author(s):  
Srdan Verstovsek ◽  
Cem Akin ◽  
Taghi Manshouri ◽  
Alfonso Quintás-Cardama ◽  
Ly Huynh ◽  
...  
Keyword(s):  
Mast Cells ◽  
Tyrosine Kinase ◽  
Tyrosine Kinase Inhibitor ◽  
Kinase Inhibitor ◽  
Wild Type ◽  
Human Mast Cells

Efficacy of Ponatinib Against ABL Tyrosine Kinase Inhibitor Resistant Leukemia Cells

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4413-4413
Author(s):  
Seiichi Okabe ◽  
Tetsuzo Tauchi ◽  
Kazuma Ohyashiki
Keyword(s):  
Cell Proliferation ◽  
Tyrosine Kinase ◽  
Tyrosine Kinase Inhibitor ◽  
Cell Line ◽  
Cell Lines ◽  
Kinase Inhibitor ◽  
Resistant Cell ◽  
Point Mutant ◽  
Abl Tyrosine Kinase ◽  
Abl Tyrosine Kinase Inhibitor

Abstract Abstract 4413 Dasatinib (Sprycel®) and nilotinib (Tasigna®) have each shown superior efficacy as front line treatment for patients with chronic myeloid leukemia (CML)-chronic phase (CP) in comparison with imatinib. Dasatinib and nilotinib are also used for the treatment of CML patients resistant or intolerant to imatinib therapy. However, a substantial number of patients are acquired resistance to nilotinib or dasatinib, the management of CML following the development of ABL tyrosine kinase inhibitor (TKI) resistance remains a challenge. Ponatinib, also known as AP24534, is an oral, the multi-targeted TKI. Ponatinib is currently being investigated in a pivotal phase 2 clinical trial (PACE trial) in patients with resistant or intolerant CML and Philadelphia positive acute lymphoblastic leukemia (Ph+ ALL). However, the molecular and functional consequences of ponatinib against ABL TKI resistant cells are not fully known. In this study, we investigated the ponatinib efficacy by using the BCR-ABL positive cell line, K562 and ABL TKI resistant (K562 imatinib resistant (K562IR), K562 nilotinib resistant (K562NR), K562 dasatinib resistant (K562DR) cells and murine Ba/F3 cell line which was transfected BCR-ABL random mutation, and established the new imatinib and nilotinib resistant Ba/F3 BCR-ABL point mutant (E334V) cells. We first examined the cell proliferation by using resistant cell lines. The proliferation of K562IR and K562 NR and K562DR did not decrease after imatinib (10 μM) or nilotinib (2 μM) or dasatinib (1 μM) treatment compared with parental cell line, K562. The BCR-ABL kinase domain mutation was not found. Point mutant Ba/F3 cell (E334V) was also highly resistant to imatinib (IC50: 15μM) and nilotinib (IC50: 7.5μM). We next examined the intracellular signaling by using these cell lines. Phosphorylation of BCR-ABL and Crk-L was not decreased by ABL TKIs in K562IR, K562NR and Ba/F3 BCR-ABL point mutant cells (E334V). We found the one of src family kinase, Lyn was activated in K562IR and K562NR cells. Co-treatment src kinase inhibitor, PP2 and imatinib or nilotinib significantly reduced the cell proliferation of K562IR and K562NR cells. We also found the phosphorylation of Lyn was reduced and poly (ADP-ribose) polymerase (PARP) was activated. We next examined the efficacy of ponatinib against imatinib and nilotinib resistant cell lines. 72 hours treatment of ponatinib exhibits cell growth inhibition against K562 (IC50: 0.02nM), K562IR (IC50: 15nM), and K562NR (IC50: 3.5nM) cells. We also found the phosphorylation of BCR-ABL, Lyn and Crk-L was reduced and PARP was activated after ponatinib treatment. We next examined the imatinib and nilotinib resistant Ba/F3 cells with point mutant (E334V). We found the cell proliferation was significantly decreased after ponatinib treatment (IC50: 3nM). We also found the phosphorylation of BCR-ABL, Crk-L was reduced and PARP was activated after ponatinib treatment. We next investigated the ponatinib activity against dasatinib resistant cells. We found K562DR cells were highly resistant to ponatinib. IC50 was 400nM. These results suggest that the expression and protein activation signatures identified in this study provide insight into the mechanism of resistance to ABL TKIs. We also demonstrate ponatinib has anti-leukemia effect by reducing ABL and Lyn kinase activity and development of ponatinib resistance and suggests that this information may be of therapeutic relevance. Disclosures: No relevant conflicts of interest to declare.

Effects of tyrosine kinase inhibitor STI571 on human mast cells bearing wild-type or mutated c-kit

2003 ◽  
Vol 31 (8) ◽  
pp. 686-692 ◽  
Author(s):  
Cem Akin ◽  
Knut Brockow ◽  
Claudio D'Ambrosio ◽  
Arnold S Kirshenbaum ◽  
Yongsheng Ma ◽  
...  
Keyword(s):  
Mast Cells ◽  
Tyrosine Kinase ◽  
Tyrosine Kinase Inhibitor ◽  
Kinase Inhibitor ◽  
Wild Type ◽  
Human Mast Cells

EXEL-0862, a novel tyrosine kinase inhibitor, induces apoptosis in vitro and ex vivo in human mast cells expressing the KIT D816V mutation

Blood ◽  
2006 ◽  
Vol 109 (1) ◽  
pp. 315-322 ◽  
Author(s):  
Jingxuan Pan ◽  
Alfonso Quintás-Cardama ◽  
Hagop M. Kantarjian ◽  
Cem Akin ◽  
Taghi Manshouri ◽  
...  
Keyword(s):  
Mast Cell ◽  
Tyrosine Kinase ◽  
Tyrosine Kinase Inhibitor ◽  
Cell Lines ◽  
Kinase Inhibitor ◽  
Human Mast Cell ◽  
Activation Loop ◽  
Dependent Manner ◽  
Juxtamembrane Domain ◽  
Dose Dependent

Abstract Gain-of-function mutations of the receptor tyrosine kinase KIT play a key role in the pathogenesis of systemic mastocytosis (SM), gastrointestinal stromal tumors (GISTs), and some cases of acute myeloid leukemia (AML). Whereas KIT juxtamembrane domain mutations seen in most patients with GIST are highly sensitive to imatinib, the kinase activation loop mutant D816V, frequently encountered in SM, hampers the binding ability of imatinib. We investigated the inhibitory activity of the novel tyrosine kinase inhibitor EXEL-0862 against 2 subclones of human mast cell line-1 (HMC-1)—HMC-1.1, harboring the juxtamembrane domain mutation V560G, and HMC-1.2, carrying V560G and the activation loop mutation D816V, found in more than 80% of patients with SM. EXEL-0862 inhibited the phosphorylation of KIT in a dose-dependent manner and decreased cell proliferation in both mast cell lines with higher activity against HMC-1.2 cells. The phosphorylation of KIT-dependent signal transducer and activator of transcription-3 (STAT3) and STAT5 was abrogated upon exposure to nanomolar concentrations of EXEL-0862. In addition, EXEL-0862 induced a time- and dose-dependent proapoptotic effect in both mast cell lines and caused a significant reduction in mast-cell content in bone marrow samples from patients with SM harboring D816V and from those without the D816V mutation. We conclude that EXEL-0862 is active against KIT activation loop mutants and is a promising candidate for the treatment of patients with SM and other KIT-driven malignancies harboring active site mutations.

scholarly journals Effect of tyrosine kinase inhibitor STI571 on the kinase activity of wild-type and various mutated c-kit receptors found in mast cell neoplasms

Oncogene ◽  
2003 ◽  
Vol 22 (5) ◽  
pp. 660-664 ◽  
Author(s):  
Yael Zermati ◽  
Paulo De Sepulveda ◽  
Frederic Féger ◽  
Sebastion Létard ◽  
Joelle Kersual ◽  
...  
Keyword(s):  
Mast Cell ◽  
Tyrosine Kinase ◽  
Tyrosine Kinase Inhibitor ◽  
Kinase Activity ◽  
Kinase Inhibitor ◽  
Wild Type

1J373 Is a Promising Agent Against C-KIT driven Acute Myeloid Leukemia

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1448-1448
Author(s):  
Hui-Jen Tsai ◽  
Tsu-An John Hsu ◽  
Chiung-Tong Chen ◽  
Weir-Torn Jiang ◽  
Hui-You Lin ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Tyrosine Kinase Inhibitor ◽  
Cell Lines ◽  
Myeloid Leukemia ◽  
Kinase Inhibitor ◽  
Leukemia Cell ◽  
Kit Mutation ◽  
Leukemia Cell Lines ◽  
Myeloid Leukemia Cell

Abstract Acute myeloid leukemia (AML) carrying t(8;21)(q22;q22) and inv(16)/t(16;16)(p13;q22) are classified as French-American-British (FAB) AML subtype M2 or monocytic with eosinophilic differentiation (M4Eo) by morphology and as core binding factor (CBF)-AML according to pathogenesis. CBF-AML accounts for approximately 15% of AML and frequently harbors c-KIT mutation (17∼46%). C-KIT mutated CBF-AML patients usually have higher baseline white blood cell count, higher relapse rate and shorter event free survival/overall survival after conventional chemotherapy than those without c-KIT mutation. It is conceived that c-KIT mutation is a crucial hit and cooperates with AML1-ETO resulting from t(8;21)(q22;q22) to cause overt AML. Imatinib, a tyrosine kinase inhibitor suppressing c-KIT activation, has been used in c-KIT mutated AML, systemic mastocytosis (SM) and gastrointestinal stromal tumor (GIST). However, c-KIT exon17 D816V mutation, a frequent mutation of CBF-AML and SM, is associated with primary imatinib resistance. 1J373, a multi-targeted tyrosine kinase inhibitor, which was initially designed as a FLT3 inhibitor but later found to target c-KIT as well. It has been shown to effectively inhibit the proliferation of FLT3-ITD mutated leukemia cell lines, MV4;11 and MOLM-13, both in vitro and in vivo. (unpublished data) Among a series of myeloid leukemia cell lines without FLT3-ITD mutation, including THP-1, HL-60, K562, KG-1, and kasumi-1, the sensitivity to 1J373 is closely associated with the presence of constitutive c-KIT activation (Figure 1A). The IC50 of 1J373 for cells with (K562, KG-1 and kasumi-1) and without (THP-1 and HL-60) constitutive c-KIT activation was below 50 nM and beyond 1000 nM, respectively. 1J373 suppressed the phosphorylation of c-KIT for cell lines with constitutively activated c-KIT, which suggested that 1J373 may suppress the proliferation of KG-1, K562, and kasumi-1 by inhibiting c-KIT (Figure 1B).We further compared the efficacy of 1J373 and imatinib in kasumi-1, a cell line with t(8;21)/AML1-ETO and c-KIT exon 17 N822K mutation. At 1000nM of concentration, the phosphorylation of c-KIT was effectively inhibited by imatinib at 2-hour but partially recovered after 8 hours; while 1J373 treatment resulted in a sustained inhibition for 24 hours. The inhibition of c-KIT activation by both agents was accompanied with corresponding changes in the phosphorylation status of its downstream signaling pathway molecules, including PI3K, AKT, mTOR, and MAPK (Figure 2). 1J373 induced cell cycle arrest of kasumi-1 at G1 phase with increase of subG1 population time-dependently and induced apoptosis of kasumi-1 through activation of caspase 8 and 9, and upregulation of proapoptotic proteins Bax and Bak. The in vivo experiments are in progress. In conclusion, 1J373, a multi-targeted tyrosine kinase inhibitor, can effectively inhibit the proliferation and induce the apoptosis of c-KIT activated leukemia cells. It has the potential to be used in clinical practice to treat c-KIT driven, particularly c-KIT mutated, AML.Figure 1c-KIT and phosphorylated c-KIT expression in myeloid leukemia cell linesFigure 1. c-KIT and phosphorylated c-KIT expression in myeloid leukemia cell linesFigure 2c-KIT and its downstream signalings expression in kasumi-1 cells treated with imatinib and 1J373Figure 2. c-KIT and its downstream signalings expression in kasumi-1 cells treated with imatinib and 1J373 Disclosures: No relevant conflicts of interest to declare.

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