WH-4-023-Dual-LCK-SRC-inhibitor-DataSheet-生命科學試劑-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEWH-4-023Cat. No.: HY-12299CAS No.: 837422-57-8Synonyms: Dual LCK/SRC inhibitor分式: CHNO分量: 568.67作靶點: Src作通路: Protein Tyrosine Kinase/RTK儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實驗 DMSO : 46.8

2、mg/mL (82.30 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲備液1 mM 1.7585 mL 8.7924 mL 17.5849 mL5 mM 0.3517 mL 1.7585 mL 3.5170 mL10 mM 0.1758 mL 0.8792 mL 1.7585 mL請根據(jù)產品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存式和期限。體內實驗 請根據(jù)您的實驗動物和給藥式選擇適當?shù)娜芙獍?，配制前請先配制澄清的儲備液，再依次添加助溶?

3、為保證實驗結果的可靠性，體內實驗的作液，建議您現(xiàn)現(xiàn)配，當天使；澄清的儲備液可以根據(jù)儲存條件，適當保存；以下溶劑前的百分 指該溶劑在您配制終溶液中的體積占)：1. 請依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 0.77 mg/mL (1.35 mM); Clear solution2. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 0.77 mg/mL (1.35 mM); Clear solution1/2 Master of Small Molecules 您邊的抑制

4、劑師www.MedChemEBIOLOGICAL ACTIVITY物活性 WH-4-023種有效的選擇性的 Lck/Src 抑制劑，IC50 分別為 2 nM/6 nM，對p38 和 KDR作效果稍弱。IC50 & Target IC50: 2 nM (Lck), 6 nM (Src) 1體外研究 WH-4-023 shows a similar potency increase on Lck as 2-substituted variants 1.PROTOCOLKinase Assay 1 The Lck HTRF kinase assay involves ATP-dependent p

5、hosphorylation of a biotinylated substrate peptide ofgastrin in the presence or absence of inhibitor compound. The final concentration of gastrin is 1.2 M. Thefinal concentration of ATP is 0.5 M (Km app =0.60.1 M), and the final concentration of Lck (a GST-kinasedomain fusion (AA 225509) is 250 pM.

6、Buffer conditions are as follows: 50 mM HEPES pH=7.5, 50 mMNaCl, 20 mM MgCl2, 5 mM MnCl2, 2 mM DTT, 0.05% BSA. The assay is quenched and stopped with 160 Lof detection reagent. Detection reagents are as follows: Buffer made of 50 mM Tris, pH=7.5, 100 mM NaCl, 3mM EDTA, 0.05% BSA, 0.1% Tween20. Prior

7、 to reading, Streptavidin allophycocyanin (SA-APC) is added ata final concentration in the assay of 0.0004 mg/mL, along with europilated anti-phosphotyrosine Ab (Eu-anti-PY) at a final conc of 0.025 nM. The assay plate is read in a Discovery fluorescence plate reader withexcitation at 320 nm and emi

8、ssion at 615 and 655 nm 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 The purpose of this assay is to test the potency of T cell receptor (TCR; CD3) and CD28 signaling pathwayinhibitors in human T cells. T cells are purified from human

9、peripheral blood lymphocytes (hPBL) andpreincubated with or without compound prior to stimulation with a combination of an anti-CD3 and an anti-CD28 antibody in 96-well tissue culture plates (1105 T cells/well). Cells are cultured for 20 h at 37C in 5%CO2 and then secreted IL-2 in the supernatants i

10、s quantified by cytokine ELISA. The cells remaining in thewells are then pulsed with 3H-thymidine overnight to assess the T cell proliferative response. Cells areharvested onto glass fiber filters and 3H-thymidine incorporation into DNA is analyzed by liquid scintillationcounter. For comparison purp

11、oses, phorbol myristic acid (PMA) and calcium ionophore are used incombination to induce IL-2 secretion from purified T cells. Potential inhibitor compounds are tested forinhibition of this response as described above for anti-CD3 and -CD28 antibodies. Human whole-blood anti-CD3/CD28-induced IL-2 se

12、cretion assays are run in a similar fashion as described above using whole bloodfrom normal volunteers diluted 50% in tissue culture medium prior to stimulation 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Martin MW, et al. Novel 2-aminopyrimidine carbamates as potent and orally active inhibitors of Lck: synthesis, SAR, and in vivoantiinflammatory activity. J Med Chem. 2006 Aug 10;49(16):4981-91.McePdfHeight2/2 Master of Small Molecules 您邊的抑制劑師www.MedChem