AR-A014418-AR-0133418-DataSheet-生命科學(xué)試劑-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEAR-A014418Cat. No.: HY-10512CAS No.: 487021-52-3Synonyms: AR 0133418; GSK 3 inhibitor VIII; AR 014418分式: CHNOS分量: 308.31作靶點(diǎn): GSK-3作通路: PI3K/Akt/mTOR; Stem Cell/Wnt儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1

2、month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 100 mg/mL (324.35 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 3.2435 mL 16.2174 mL 32.4349 mL5 mM 0.6487 mL 3.2435 mL 6.4870 mL10 mM 0.3243 mL 1.6217 mL 3.2435 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。體內(nèi)實(shí)驗(yàn) 請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥式選擇適當(dāng)

3、的溶解案，配制前請(qǐng)先配制澄的儲(chǔ)備液，再依次添加助溶劑(為保證實(shí)驗(yàn)結(jié)果的可靠性，體內(nèi)實(shí)驗(yàn)的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使；澄的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)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(qǐng)依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (8.11 mM); Clear solution1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEBIOLOGICAL ACTIVITY物活性 AR-A014418種有效，選擇性，ATP

4、競(jìng)爭(zhēng)性的 GSK3 抑制劑，IC50 值為 104 nM。IC50 & Target GSK-3104 nM (IC50)體外研究 AR-A014418 blocks the phosphorylation of tau at a GSK3-specific site (Ser-396) in 3T3 fibroblastsexpressing human four-repeat tau protein, with an IC50 of 2.7 M, and protects cultured N2A cells fromdeath cuased by PI3K/PKB pathway blo

5、ckage. AR-A014418 also shows inhibitory effect onneurodegeneration mediated by beta-amyloid peptide in hippocampal slices 1. AR-A014418 decreasesneuroendocrine markers and suppresses neuroblastoma cell growth in NGP and SH-5Y-SY cells 2.體內(nèi)研究 AR-A014418 (0-4 mg/kg, i.p.) delays the onset of symptoms,

6、 enhances motor activity, blocks diseaseprogression, and postpons the endpoint of the disease in ALS mouse model with the G93A mutant humanSOD1 3. Furthermore, AR-A014418 suppresses acetic acid- and formalin-induced nociception in mice viamodulating NMDA and metabotropic receptor signaling as well a

7、s TNF- and IL-1 transmission in the spinalcord 4.PROTOCOLKinase Assay 1 The competition experiments are carried out in duplicate with 10 concentrations of the inhibitor in clear-bottomed microtiter plates. The biotinylated peptide substrate, biotin-AAEELDSRAGS(PO3H2)PQL, is addedat a final concentra

8、tion of 2 M in an assay buffer containing 6 milliunits of recombinant human GSK3 (equalmix of both and ), 12 mM MOPS, pH 7.0, 0.3 mM EDTA, 0.01% -mercaptoethanol, 0.004% Brij 35, 0.5%glycerol, and 0.5 g of bovine serum albumin/25 L and preincubated for 10-15 min. The reaction is initiatedby the addi

9、tion of 0.04 Ci of -33PATP and unlabeled ATP in 50 mM Mg(Ac)2 to a final concentration of 1M ATP and assay volume of 25 L. Blank controls without peptide substrate are used. After incubation for20 min at room temperature, each reaction is terminated by the addition of 25 L of stop solution containin

10、g 5mM EDTA, 50 M ATP, 0.1% Triton X-100, and 0.25 mg of streptavidin-coated SPA beads corresponding toappr 35 pmol of binding capacity. After 6 h the radioactivity is determined in a liquid scintillation counter.Inhibition curves are analyzed by non-linear regression using GraphPad Prism.MCE has not

11、 independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Cell viability is assessed by calcein/propidium iodide uptake. Calcein AM is taken up and cleaved byesterases present within living cells, yielding yellowish-green fluorescence, whereas PI is only taken

12、up bydead cells, which become orange-red fluorescent. In brief, N2A cells are cultured for 2 days in vitro and thentreated with 50 M LY-294002 in the presence of AR-A014418 or vehicle (DMSO) for 24 h. Subsequently,N2A cells are incubated for 30 min with 2 M PI and 1 M calcein-AM. The cultures are th

13、en rinsed threetimes with Hanks buffered saline solution containing 2 mM CaCl2, and the cells are visualized byfluorescence microscopy using a Zeiss Axiovert 135 microscope. Three fields (selected at random) areanalyzed per well (appr 300 cells/field) in at least three different experiments. Cell de

14、ath is expressed aspercentage of PI-positive cells from the total number of cells. In every experiment, specific cell death isobtained after subtracting the number of dead cells present in vehicle-treated cultures.2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEMCE has not independently confirmed t

15、he accuracy of these methods. They are for reference only.Animal First, to examine the effects of GSK-3 inhibition on the clinical symptoms, life span, and motor behaviorAdministration 3 function of ALS, 56 Tg mice are divided into four groups. In each group, 0.5 mL of normal saline is mixed witheit

16、her 0 g (control group), 1 g (group A), 2 g (group B) or 4 g (group C) of AR-A014418 per gram ofmouse, and injected intraperitoneally into 14 animals per group 5 days a week beginning 60 days after birth.The mice are sacrificed at the endpoint described below.MCE has not independently confirmed the

17、accuracy of these methods. They are for reference only. Cell Syst. 2018 Apr 25;6(4):424-443.e7. J Exp Clin Cancer Res. 2018 Jun 25;37(1):122. J Cell Physiol. 2019 Jul;234(7):10411-10420. Toxicol Sci. 2018 Apr 1;162(2):475-487.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Bhat

18、 R, Xue Y, Berg S, Structural insights and biological effects of glycogen synthase kinase 3-specific inhibitor AR-A014418. J BiolChem. 2003 Nov 14;278(46):45937-45.2. Carter YM, et al. Specific glycogen synthase kinase-3 inhibition reduces neuroendocrine markers and suppresses neuroblastoma cellgrowth. Cancer Biol Ther. 2014 May;15(5):510-5.3. Koh SH, et al. Inhibition of glycogen synthase kinase-3 suppresses the onset of symptoms and disease progression of G93A-SOD1mouse model of ALS. Exp Neurol. 2007 Jun;205(2):336-464. Martins DF, et al. The antinociceptive effects of AR-A014418, a selecti