Dasatinib

Dasatinib is a novel, potent and multi-targeted inhibitor that targets Abl, Src and c-Kit, with IC50 of <1 nM, 0.8 nM and 79 nM in cell-free assays, respectively.

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Dasatinib Chemical Structure

Dasatinib Chemical Structure
Molecular Weight: 488.01

Validation & Quality Control

Product Use Citation(59)

Customer Product Validation(10)

Quality Control & MSDS

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Product Information

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  • Research Area
  • Combination Therapy
    Combination Therapy

Product Description

Biological Activity

Description Dasatinib is a novel, potent and multi-targeted inhibitor that targets Abl, Src and c-Kit, with IC50 of <1 nM, 0.8 nM and 79 nM in cell-free assays, respectively.
Targets Abl [1]
(Cell-free assay)
Src [1]
(Cell-free assay)
c-Kit (D816V) [2]
(Cell-free assay)
c-Kit (wt) [2]
(Cell-free assay)
IC50 0.6 nM 0.8 nM 37 nM 79 nM
In vitro Dasatinib is more effective than imatinib in inhibiting the proliferation of Ba/F3 cells expressing wild-type Bcr-Abl and Bcr-Abl mutants, with the exception of T315I. Dasatinib has a two-log (∼325-fold) increased potency relative to imatinib. Dasatinib potently inhibits wild-type Abl kinase and all mutants except T315I over a narrow range. Dasatinib directly targets wild-type and mutant Abl kinase domains and inhibits autophosphorylation and substrate phosphorylation in a concentration-dependent manner. Dasatinib displays 325-fold greater potency compared with imatinib against cells expressing wild-type Bcr-Abl. [1] The percent of colonies of TgE bone marrow cells are decreased from 100% in untreated wells to 4.12% in Dasatinib treated wells. In the presence of Dasatinib, the difference in the percentage of colonies formed by WT and TgE bone marrow cells is statistically significant. Expression of LMP2A is able to promote B lymphocyte survival and proliferation, which can be inhibited by targeting Lyn and/or c-Abl kinases through Dasatinib. [3] Dasatinib treatment inhibits Src signaling, decreases growth, and induces cell cycle arrest and apoptosis in a subset of thyroid cancer cells. Treatment with increasing doses of Dasatinib (0.019 μM to 1.25 μM) for 3 days inhibits the growth of the C643, TPC1, BCPAP, and SW1736 cell lines by about 50% at low nanomolar concentrations, while higher concentrations are required to inhibit the growth of the K1 cell line. Treatment with 10 nM or 50 nM Dasatinib results in a 9-22% increase of cells in the G1 population among BCPAP and SW1736 and K1 cells, and a corresponding 7-18% decrease in the percentage of cells in the S phase. [4]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity Description
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EVSA-TMkLVS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?MVHJR|UxRTlwMke3PVMh|ryP
GT3TKBMXrHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?MnqyTWM2OD17LkO1OVQ3KM7:TR?=
NB6MVPHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?NFnxV|VKSzVyPUmuPVIzPTlizszN
EHEBNEnUbYlIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=NF\tUGdKSzVyPUGwMlA3PTZizszN
HELNHi0bXRIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=MmHSTWM2OD1zMD60O|c3KM7:TR?=
ALL-PONX63XGVIT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NXq1b4d2UUN3ME2xNE44QTN6IN88US=>
TGWMYfHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?NHHWb|JKSzVyPUGxMlI5OjhizszN
BC-3MlroS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?M{mzUGlEPTB;MUKuNVE{QCEQvF2=
IA-LMMn3LS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?MojsTWM2OD1zMj60OFQ2KM7:TR?=
UACC-257NVe4[2JtT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=M1GzUWlEPTB;MUKuPVE6QCEQvF2=
KP-N-YSMWPHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?MoHkTWM2OD1zMj65Nlg{KM7:TR?=
RajiMUHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?Mn3DTWM2OD1zMz63OFk4KM7:TR?=
SF539MV3Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?MULJR|UxRTF|Lki1OVch|ryP
DMS-153NFjkU3dIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=MlHKTWM2OD1zND6wNFI5KM7:TR?=
L-540MmPwS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?NUXVeYNUUUN3ME2xOU4xPjd{IN88US=>
MN-60MVHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?NIf1[XRKSzVyPUG1MlE6PzlizszN
RPMI-8866M3;rbGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7NUPJR49vUUN3ME2xO{41PDV2IN88US=>
NCI-H510AMVvHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?NIfuT4pKSzVyPUG5MlM6PzNizszN
NB13NXnMXXFJT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NF3HUpJKSzVyPUG5MlQ5PzdizszN
HAL-01MV;Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?MkHxTWM2OD1zOT63OVQ{KM7:TR?=
NCI-H720MVrHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?MoPNTWM2OD1{MD6yO|M{KM7:TR?=
REHNXXPVGVLT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NF\a[|RKSzVyPUKwMlY{PTdizszN
KNS-81-FDNXP1NW9KT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NXLhd3RtUUN3ME2yN{4yPDZizszN
HC-1NXryO5Y4T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NIjRUopKSzVyPUK0MlU2PTFizszN
NCI-H2141MnLFS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?NW\USnpoUUN3ME2yOE44PzV2IN88US=>
MOLT-4MWnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?MkLMTWM2OD1{Nj62O|U{KM7:TR?=
OMC-1NYjoOGZFT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=MnHpTWM2OD1{Nz6xOFIzKM7:TR?=
LC-1FMUPHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?MljhTWM2OD1{Nz6zNlQ2KM7:TR?=
NCI-H1304NHLlZYpIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=Mkj3TWM2OD1{OD6xOlI5KM7:TR?=
BC-1NWXET4NNT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=M{LBT2lEPTB;MkiuOlUyKM7:TR?=
NCI-H64NGPlb4dIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=MV7JR|UxRTJ7Lk[yOVMh|ryP
MOLT-16MXjHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?MVPJR|UxRTJ7Lk[yPVIh|ryP
U-87-MGNGHt[XFIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=MULJR|UxRTNyLke2OkDPxE1?
GAKMYDHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?M1HIcGlEPTB;M{GuNlY5PiEQvF2=
ES8MnW4S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?MoXDTWM2OD1|Mj6xNlUzKM7:TR?=
HCC1599NVX1VHdvT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NYfjTpFHUUN3ME2zNk4{OzJ3IN88US=>
EB-3Mo\KS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?NX3sNm5tUUN3ME2zOE4{OTF5IN88US=>
HCC1187M3\Q[Wdzd3e2aDDJcohq[mm2aX;uJGF{e2G7NIXDSVFKSzVyPUO1MlgxPTJizszN
SK-PN-DWM4i4dWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7NGm0Z41KSzVyPUO2MlE6PDNizszN
JVM-3Mo\6S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?NHW2SpRKSzVyPUO3MlI{OzhizszN
HCC2157M2O3b2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7NFXTXGxKSzVyPUO3Mlk6PDZizszN
A4-FukNYD1NY9QT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=MUHJR|UxRTN6LkGwNFkh|ryP
COR-L279MYHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?NVXmTFFVUUN3ME20NE4zQDVzIN88US=>
DELM1HXbWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7NX7RfY5ZUUN3ME20NU46ODh4IN88US=>
NCI-H1395NEHY[|dIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=NEPBUYRKSzVyPUSyMlAyPjNizszN
MHH-NB-11NXTBd3JKT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=MXzJR|UxRTR|LkC4NVgh|ryP
NCI-H2107MU\Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?NV22VI1nUUN3ME20N{41QDR4IN88US=>
NEC8NEnUTJRIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=NEXwfoZKSzVyPUS0MlM{PiEQvF2=
COLO-684NWHNT2R{T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=MVrJR|UxRTR4LkKyOVgh|ryP
LS-411NMWjHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?Mkj0TWM2OD12OD60O|Q5KM7:TR?=

... Click to View More Cell Line Experimental Data

In vivo Dasatinib reverses splenomegaly in LMP2A/MYC double transgenic mice. Dasatinib specifically prevents colony formation by LMP2A expressing bone marrow B cells and decreased spleen size in the TgE mice. Spleen mass is significantly decreased among Dasatinib treated Tg6/λ-MYC mice when compared to the control group. Dasatinib inhibits lymphadenopathy in LMP2A/MYC double transgenic mice. Dasatinib reverses splenomegaly in Rag1KO mice engrafted with tumor cells from LMP2A/MYC double transgenic mice. Dasatinib therapy inhibits Lyn phosphorylation in B lymphocyte tumors expressing LMP2A. [3]
Features

Protocol(Only for Reference)

Kinase Assay:

[1]

Kinase autophosphorylation assays Kinase assays using wild-type and mutant glutathione S-transferase (GST)-Abl fusion proteins (c-Abl amino acids 220-498) are done. GST-Abl fusion proteins are released from glutathione-Sepharose beads before use; the concentration of ATP is 5 μM. Immediately before use in kinase autophosphorylation and in vitro peptide substrate phosphorylation assays, GST-Abl kinase domain fusion proteins are treated with LAR tyrosine phosphatase. After 1-hour incubation at 30 °C, LAR phosphatase is inactivated by addition of sodium vanadate (1 mM). Immunoblot analysis comparing untreated GST-Abl kinase to dephosphorylated GST-Abl kinase is routinely done using phosphotyrosine-specific antibody 4G10 to confirm complete (>95%) dephosphorylation of tyrosine residues and c-Abl antibody CST 2862 to confirm equal loading of GST-Abl kinase. The Dasatinib concentration range is extended to 1,000 nM for mutant T315I. These same inhibitor concentrations are used for the in vitro peptide substrate phosphorylation assays. The three inhibitors are tested over these same concentration ranges against GST-Src kinase and GST-Lyn kinase.

Cell Assay:

[1]

Cell lines Ba/F3 cell lines
Concentrations ~32 nM
Incubation Time 72 hours
Method

Ba/F3 cell lines are seeded in triplicate and incubated with escalating concentrations of Dasatinib for 72 hours. Proliferation is measured using a methanethiosulfonate-based viability assay. IC50 and IC90 values are reported as the mean of three independent experiments done in quadruplicate. The inhibitor concentration ranges are 0 nM to 32 nM (Dasatinib). The Dasatinib concentration range is extended to 200 nM for mutant T315I.

Animal Study:

[3]

Animal Models EμLMP2A (TgE and Tg6 strains), MYC (λ-MYC), and LMP2A/λ-MYC double transgenic mice (Tg6/λ-MYC)
Formulation DMSO
Dosages 30 mg/kg
Administration Administered via i.p.

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDog
Weight (kg)0.020.151.80.40.0810
Body Surface Area (m2)0.0070.0250.150.050.020.5
Km factor36128520
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)
1

References

[1] O'Hare T, et al. Cancer Res. 2005, 65(11), 4500-4505.

[2] Shah NP, et al. Blood, 2006, 108(1), 286-291.

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Clinical Trial Information( data from http://clinicaltrials.gov, updated on 2015-06-27)

NCT Number Recruitment Conditions Sponsor
/Collaborators
Start Date Phases
NCT02420717 Not yet recruiting Leukemia M.D. Anderson Cancer Center|Incyte Corporation August 2015 Phase 2
NCT02389309 Not yet recruiting Solid Tumors M.D. Anderson Cancer Center June 2015 Phase 1
NCT02297139 Not yet recruiting Multiple Indications Cancer Bristol-Myers Squibb June 2015 Phase 2
NCT02428855 Not yet recruiting Cholangiocarcinoma Massachusetts General Hospital|Bristol-Myers Squibb|Targe  ...more Massachusetts General Hospital|Bristol-Myers Squibb|TargetCancer Foundation April 2015 Phase 2
NCT02326311 Not yet recruiting Leukemia, Chronic Myeloid Azienda Ospedaliera Spedali Civili di Brescia January 2015 Phase 3

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Chemical Information

Download Dasatinib SDF
Molecular Weight (MW) 488.01
Formula

C22H26ClN7O2S

CAS No. 302962-49-8
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms BMS-354825
Solubility (25°C) * In vitro DMSO 98 mg/mL (200.81 mM)
Water <1 mg/mL (<1 mM)
Ethanol <1 mg/mL (<1 mM)
In vivo 1% DMSO/30% polyethylene glycol/1% Tween 80 30 mg/mL
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Name N-(2-chloro-6-methylphenyl)-2-(6-(4-(2-hydroxyethyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide

Customer Product Validation (10)


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Rating
Source Cell Res 2011 21, 1080-1087. Dasatinib purchased from Selleck
Method Microangiography
Cell Lines embryos
Concentrations 30 μM
Incubation Time
Results MEK1/2 may cooperate with VEGFR to regulate blood vessel lumen diameter. We examined this hypothesis by combining VEGFR inhibitor Sunitinib and MEK1/2 inhibitor U0126. Indeed, U0126 (10 µM) combined with Sunitinib (20 µM) resulted in a reduction of vessel lumen size (Figure F), whereas individually they did not. Combined treatment of another MAP kinase signaling pathway inhibitor Dasatinib (20 µM) with Sunitinib (20 µM) produced a similar phenotype as PP1, whereas each individual inhibitor did not result in any vessel shrinkage even at much higher concentrations ( Figure C).Since there are three major VEGF receptors, it is desirable to determine which receptor is involved in combinatory action with MEK1/2. To address this issue, additional highly selective VEGFR inhibitors PTK787 and ZM323881 were tested. Both of these inhibitors (PTK787 or ZM323881), when individually combined with Dasatinib or U0126, induced a reduction of vessel lumen size (Figure D, E, G, and H).

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Rating
Source Clin Cancer Res 2011 17, 762-770. Dasatinib purchased from Selleck
Method Assessment of cell viability and apoptosis
Cell Lines B-CLL patient leukemic cells
Concentrations 2-10 μM
Incubation Time 48 h
Results The Dasatinib+Nutlin-3 combination promotes synergistic cytotoxicity in primary CLL cells as well as in p53 wild-type and p53 deleted/ mutated leukemic cell lines.

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Rating
Source Clin Cancer Res 2011 17, 762-770. Dasatinib purchased from Selleck
Method Western blot
Cell Lines Leukemic cell lines
Concentrations 10 μM
Incubation Time 24 h
Results Dasatinib marginally affected both the basal levels of p53 and the accumulation of p53 induced by Nutlin-3 in the p53 wild -type cells (Fig. A).Interestingly, however, levels of both MDM2 and p21 proteins, 2 of the best characterized transcriptional targets of p53, weres significantly (P < 0.05) decreased in cells treated with Dasatinib+ Nutlin-3 with respect to cells treated with Nutlin-3 alone (Fig. B).

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Rating
Source Clin Cancer Res 2011 17, 762-770. Dasatinib purchased from Selleck
Method Phospho-kinase array analysis
Cell Lines EHEB cell lines/BJAB cell lines
Concentrations 10 μM
Incubation Time 16 h
Results Although Nutlin-3 had no effects on these pathways, Dasatinib alone as well as Dasatinib+Nutlin-3 markedly inhibited the basal phosphorylation of ERK1/2, p38/MAPK, and Akt . Of interest, the combination of Dasatinib+Nutlin-3 selectively enhanced (P < 0.05) the inhibition of Akt phosphorylation with respect to Dasatinib alone, as validated by Western blot analysis.

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Rating
Source Biochem Pharmacol 2011 82, 1457-1466. Dasatinib purchased from Selleck
Method MTT assay
Cell Lines MOLM-13 cells, HL-60 cells
Concentrations 1-15 μM
Incubation Time 24-72 h
Results Dasatinib and other TKI decreased the viability of the two myeloid cell lines in a dose- and time-dependent manner. Nevertheless, the anti-leukemic efficacy of Dasatinib and other TKI varied considerably in both cell lines.

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Rating
Source Biochem Pharmacol 2011 82, 1457-1466. Dasatinib purchased from Selleck
Method Evaluation of differentiation
Cell Lines MOLM-13 cells, HL-60 cells
Concentrations 5 μM
Incubation Time
Results In a first series of experiments, we assessed four signs of morphological differentiation, May-Gruenwald-Giemsa staining of MOLM-13 cells demonstrated that dasatinib induced a dose-dependent decrease in cytoplasmic basophilia and in the nucleo-cytoplasmic ratio. Dasatinib also led to the appearance of dented nuclei and cytoplasmic granulation( Fig.A and B).In HL-60 cells,dasatinib(5 μM) once more exhibited the highest differentiation-inducing capacity. Thus dasatinib diminished basophilia of the cytoplasm concomitantly with the nucleo-cytoplasmic ratio, and induced the appearance of nuclear lobulation and cytoplasmic granules(Fig. G). Next, we determined the degree of CD11b expression, a marker indicating myeloid differentiation, in TKI-treated MOLM-13 dasatinib exhibited the most pronounced capacity to increase CD11b surface expression( Fig. C and D).To corroborate the TKI-driven differentiation, we quantified the enzymatic activity of alkaline phosphatase by means of the NBT-reduction assay after an incubation period of 6 days in MOLM-13 cells.,Once again, the NBT-reductive activity was highest in cells treated with dasatinib, which was as active as the positive control, ATRA( Fig. E and F)

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Rating
Source Invest New Drugs 2010 30, 417-422. Dasatinib purchased from Selleck
Method Assessment of cell viability
Cell Lines leukemic cells
Concentrations 10 μM
Incubation Time 24/48 h
Results Dasatinib dose-dependently reduced cell viability in all leukemic cell lines, with variable efficacy irrespect ively of the p53 status. In particular, when used at 10 μM, Dasatinib induced a progressive decrease of the total number of viable cells ranging from approximately 30% (JVM-2 and MEC-2) to 50% (EHEB and BJAB) after 48 h of treat ment (Fig.a). In order to investigate whet her the decreased leukemic cell viability was mainly due to either cytotoxic or cytostatic activity, we have analyzed the effects of Dasatinib on apoptosis,As shown in Fig. b, exposure to Dasatinib induced a significant (p < 0.05) increase of apoptosis in all leukemic cell lines, as evaluated by Annexin-V/PI staining at 48 h of treatment.

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Rating
Source Invest New Drugs 2010 30, 417-422. Dasatinib purchased from Selleck
Method phospho-kinase array analysis
Cell Lines EHEB (p53wt) cell lines, BJAB (p53mut) B cell lines
Concentrations
Incubation Time 24 h
Results As shown in Fig. a , the most evident effects of Dasatinib in both cell models were represented by a significant down-regulation of the phosphorylation levels of p38 MAPK, ERK1/2 and CREB. In addition, densitometric analysis of the phospho-kinase array demonstrated that Dasatinib also inhibited the phosphorylation levels of several STAT family members (STAT1, STAT2, STAT3, STAT5a/b and STAT6), the most evident effect in both p53 wild-type (EHEB) and p53 mutated (BJAB) cell lines being on STAT2, STAT3 and STAT6 (Fig. b)

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Rating
Source Invest New Drugs 2010 30, 417-422. Dasatinib purchased from Selleck
Method Western blot
Cell Lines p53wild-type(EHEB, JVM-2) cell lines, p53mutated(MEC-2, BJAB) cell lines
Concentrations 10 μM
Incubation Time 24 h
Results "Upon exposure to Dasatinib, a decrease of phosphorylated proteins was confirmed by Western blot analys is in all cell lines investigated without significant differences between p53wild-type and p53mutated cell lines.

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Rating
Source Dr. Thomas Kruwel of Fraunhofer-Institute for Toxicology and Experimental Medicine-Dasatinib (BMS-354825) purchased from Selleck. Dasatinib purchased from Selleck
Method Cell vability assays
Cell Lines A2C12, Beta D5, Gamma A3, Gamma D12, A549, CaCo2, HepG2 cell lines
Concentrations 0.001-1000 nM
Incubation Time 24/96 h
Results

Product Use Citation (59)

Tech Support & FAQs

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Tel: +1-832-582-8158 Ext:3

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