A-419259-trihydrochloride-RK-20449-trihydrochloride-DataSheet-生命科學(xué)試劑-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEA 419259 trihydrochlorideCat. No.: HY-15764ACAS No.: 1435934-25-0Synonyms: RK 20449 trihydrochloride分式: CHClNO分量: 592作靶點(diǎn): Src作通路: Protein Tyrosine Kinase/RTK儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶

2、解性數(shù)據(jù)體外實(shí)驗(yàn) H2O : 55 mg/mL (92.91 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 1.6892 mL 8.4459 mL 16.8919 mL5 mM 0.3378 mL 1.6892 mL 3.3784 mL10 mM 0.1689 mL 0.8446 mL 1.6892 mL請(qǐng)根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲(chǔ)備液，并請(qǐng)注意儲(chǔ)備液的保存式和期限。BIOLOGICAL ACTIVITY物活性 A 419259

3、trihydrochloride是Src家族激酶抑制劑，對(duì) Src，Lck 和Lyn 的 IC50 分別為9 nM，3 nM，3 nM。IC50 & Target IC50: 9 nM (Src), 1體外研究A 419259 Trihydrochloride is a second-generation pyrrolo-pyrimidine designed to enhance selectivity towards1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEthe Src family relative to other cytopl

4、asmic tyrosine kinases. A-419259 inhibits K-562 cells with an IC50between 0.1 and 0.3 M, and Meg-01 proliferation with an IC50 of approximately 0.1 M. A-419259 alsopotently induces apoptosis in K-562 cells beginning at 0.1 M and increasing in a dose-dependent manner.PP2 inhibits Src kinase autophosp

5、horylation in both Ph+ cell lines (K-562 and Meg-01) with an IC50 between3 and 10 M, while A-419259 blocks kinase activation between 0.1 and 0.3 M. A-419259 strongly inhibitsDAGM/Bcr-Abl cell proliferation in the absence of IL-3 with an IC50 between 0.1 and 0.3 M 1. A-419259 isa broad-spectrum pyrro

6、lo-pyrimidine inhibitor, and blocks proliferation and induces apoptosis in CML celllines. A-419259 inhibits overall SFK activity in K562 and other CML cell lines with an IC50 value of 0.1-0.3 M 2.PROTOCOLKinase Assay 1 In vitro kinase assays are performed on His(6)-tagged Lck (residues 62-509) and f

7、ull-length c-Abl purifiedfrom Sf-9 cells, and commercial sources of Lyn, Src and PKC. Lck, Lyn, Src and Abl activities are measuredwith an ELISA-based assay. Flat bottom 96-well ELISA plates are incubated with a 200 g/mL solution ofPoly(Glu,Tyr) 4 : 1 substrate in phosphate buffered saline (PBS) for

8、 1 h at 37C and then washed with PBScontaining 0.1% Tween-20 (PBS-T). Inhibitor dilutions are added to the washed plates already containing theappropriate enzyme in kinase assay buffer (250 mM Mopso, pH 6.75, 10 mM MgCl2, 2 mM MnCl2, 2.5 mMDTT, 0.02% BSA, 2 mM Na3VO4, 5% DMSO, 5 M ATP). After incuba

9、tion at room temperature for 20 min,plates are washed three times with PBS-T and plate-bound phosphotyrosine is detected with an anti-phosphotyrosine-HRP antibody conjugate and subsequent color development with K-Blue reagents. Allassays are optimized to use the least amount of enzyme necessary for

10、a reproducible signal-to-noise ratio1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 K-562 cells are grown in RPMI 1640 supplemented with 10% fetal calf serum (FCS), and 50 g/mLGentamycin. Meg-01 cells are cultured in Vitacell modified RPM

11、I 1640 (ATCC), supplemented with 10% FCSand 50 g/mL Gentamycin. The human GM-CSF-dependent myeloid leukemia cell line TF-1 is grown inRPMI 1640 supplemented with 10% FCS, 50 g/mL Gentamycin, and 1 ng/mL of recombinant human GM-CSF. DAGM murine myeloid leukemia cells are cultured in RPMI 1640 supplem

12、ented with 10% FCS, 50 g/mL Gentamycin, and 0.5 ng/mL recombinant IL-3. Concentrated stock solutions of PP2 (5 mM) and A-419259 (2 mM) are prepared in DMSO and stored at -20C. Cellular proliferation is measured using theLive/Dead growth assay. This assay employs calcein-AM, a fluorogenic esterase su

13、bstrate that is taken upby viable cells and hydrolyzed intracellularly, releasing a green fluorescent product. Briefly, 104 cells areplated per well in 96-well plates for each day of a 4-day time course. Various concentrations of PP2, A-419259 or vehicle control are added to the wells (five wells pe

14、r concentration per day) and the plates areincubated at 37C. At each time point, one plate is centrifuged at 1500 g for 10 min to pellet the cells. Cellsare washed with phosphate buffered saline (PBS), and calcein-AM is added to each well to a finalconcentration of 1 M. Plates are incubated in the d

15、ark at room temperature for 1 h. The plates are then readat 485/530 nm (excitation/emission) using a SpectraMax Gemini XS fluorescent plate reader and data areanalysed with SoftMax Pro software 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.2/3 Maste

16、r of Small Molecules 您邊的抑制劑師www.MedChemE戶(hù)使本產(chǎn)品發(fā)表的科研獻(xiàn) Blood. 2016 Jun 23;127(25):3237-52. Patent. US20170333436A1.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Wilson MB, et al. Selective pyrrolo-pyrimidine inhibitors reveal a necessary role for Src family kinases in Bcr-Abl signal transduction andoncogenesis. Oncogene. 2002 Nov 21;21(53):8075-88.2. Pene-Dumitrescu T, et al. An inhibitor-resistant mutant of Hck protects CML cells against the antiproliferative and apoptotic effects of thebroad-spectrum Src family ki