GW843682X-GW843682-DataSheet-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEGW843682XCat. No.: HY-11003CAS No.: 660868-91-7Synonyms: GW843682分式: CHFNOS分量: 477.46作靶点: Polo-like Kinase (PLK)作通路: Cell Cycle/DNA Damage储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 DMSO : 33.

2、33 mg/mL (69.81 mM; Need ultrasonic)H2O : 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (5.24 mM); Suspended solution; Need ultrasonic2. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (5.24 mM); Clear solution1/3 Master of Small Molecules 您边的抑制剂师www.MedChemEBIOLOGICAL ACTIVITY物活性 GW8

3、43682X种选择性的，ATP-竞争性的 PLK1 and PLK3 抑制剂，IC50 值分别为 2.2 nM and 9.1 nM，选择性是其他 30 种激酶的 100 多倍。IC50 & Target PLK1 PLK3 PDGFR1 VEGFR22.2 nM (IC50) 9.1 nM (IC50) 160 nM (IC50) 360 nM (IC50)Aurora A CDK2/cyclin A4800 nM (IC50) 7600 nM (IC50)体外研究 GW843682X (compound 1) is effective on inhibition of growth of

4、tumor cells, with IC50s of 0.41, 0.57, 0.11,0.38, and 0.70 M for A549, BT474, HeLa, H460 and HCT116 cell lines. GW843682X dose-dependentlyinhibits PLK1 phosphorylation of Ser15-p53, with an IC50 of 0.14 M. GW843682X (3 M) causes a strongG2-M arres in HDF cells and H460 cells after treatment for 24,

5、48, and 72 h. GW843682X (5 M) leads toapoptosis in H460 cells instead of HDF cells 1. GW843682X inhibits proliferation of U937 cells with anEC50 of 120 nM. GW843682X (500 nM) in combination with 5 M VP-16 suppresses 50% of entry intomitosis in U937 cells 2. GW843682X (0.06-1M) has inhibitory activit

6、ies against proliferation of acuteleukemia cells, and potentiates the anti-proliferative activity of vincristine. Moreover, GW843682X (0.1-1M)induces apoptosis of leukemia cells in a dose- and time-dependent manner. GW843682X (0.5-1M)dephosphorylates Bcl-xl in leukemia cells 3.PROTOCOLKinase Assay 1

7、 PLK1 and PLK3 proteins are prepared from baculovirus-infected Trichoplusia ni cells. Enzyme activity forPLK1 and PLK3 is determined as follows. All measurements are obtained under conditions where signalproduction increased linearly with time and enzyme. Test compounds are added to white 384-well a

8、ssayplates (0.1 L for 10 L and some 20 L assays, 1 L for some 20 L assays) at variable knownconcentrations in 100% DMSO. DMSO (1-5% final) and EDTA (65 mM) are used as controls. Reaction Mixcontaines the following components at 22C: 25 mM HEPES (pH 7.2); 15 mM MgCl2; 1 M ATP; 0.05 Ci/well -33PATP (1

9、0 Ci/mmol); 1 M substrate peptide (Biotin-Ahx-SFNDTLDFD); 0.15 mg/mL bovineserum albumin; 1 mM DTT; and 2 nM PLK1 kinase domain or 5 nM full-length PLK3. Reaction Mix (10 or 20L) is quickly added to each well immediately following addition of enzyme via automated liquid handlers andincubated for 1 t

10、o 1.5 h at 22C. The 20 L enzymatic reactions are stopped with 50 L of stop mix 50 mMEDTA, 4.0 mg/mL streptavidin SPA beads in Dulbeccos PBS (without Mg2+ and Ca2+), 50 M ATP perwell. The 10 L reactions are stopped with 10 L of stop mix 50 mM EDTA, 3.0 mg/mL streptavidin-coupledSPA Imaging Beadsin Du

11、lbeccos PBS (without Mg2+ and Ca2+), 50 M ATP per well. Plates are sealed,spun at 500 g for 1 min or settled overnight, and counted in Packard TopCount for 30 s/well or imaged witha Viewlux imager. Signal above background (EDTA controls) is converted to percent inhibition relative to thatobtained in

12、 control (DMSO-only) wells 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Assays are carried out and data analyzed. In these assays, H460 cells are plated at a density of 2,000 per2/3 Master of Small Molecules 您边的抑制剂师www.MedChemEwell, HD

13、F cells are plated at 5,000 per well, and the drug-resistant cell line MES-SA/DX5 and its sensitiveparent line MES-SA are plated at 7,000 and 6,000 per well, respectively, in a 96-well plate. These densitiesallowed vehicle controls to grow logarithmically during the course of the 3-day assay. All ce

14、lls are exposed to3-fold dilutions of the compound (30-0.00152 M) in low-glucose DMEM containing 5% FBS, 50 g/mLgentamicin, and 0.3% (v/v) DMSO (HDF cells); RPMI 1640 containing 5% FBS, 50 g/mL gentamicin, and0.3% (v/v) DMSO (H460); or McCoys 5A containing 5% FBS, 50 g/mL gentamicin, and 0.3% (v/v)

15、DMSO(MES-SA and MES-SA/DX5) 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Sci Rep. 2017 Aug 17;7(1):8629. Harvard Medical School LINCS LIBRARYSee more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Lansing TJ, et al. In vit

16、ro biological activity of a novel small-molecule inhibitor of polo-like kinase 1. Mol Cancer Ther. 2007 Feb;6(2):450-9. Epub 2007 Jan 31.2. Didier C, et al. Evaluation of Polo-like Kinase 1 inhibition on the G2/M checkpoint in Acute Myelocytic Leukaemia. Eur J Pharmacol.2008 Sep 4;591(1-3):102-5.3. Ikezoe T, et al. A novel treatment strategy targeting polo-like kinase 1 in hematological ma