1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemECHIR-99021 trihydrochlorideCat. No.: HY-10182BCAS No.: 1782235-14-6Synonyms: CT99021 trihydrochloride分式: CHClN分量: 574.72作靶點(diǎn): GSK-3; Autophagy作通路: PI3K/Akt/mTOR; Stem Cell/Wnt; Autophagy儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn solven

2、t -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 32 mg/mL (55.68 mM)H2O : 19 mg/mL (33.06 mM; Need ultrasonic and warming)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 1.7400 mL 8.6999 mL 17.3998 mL5 mM 0.3480 mL 1.7400 mL 3.4800 mL10 mM 0.1740 mL 0.8700 mL 1.

3、7400 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)?shù)娜芙獍?，配制前?qǐng)先配制澄清的儲(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；澄清的儲(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: 2.58 mg/mL (4.49 mM); Clear solution2. 請(qǐng)依序添加每種溶劑： 10%

4、DMSO 90% (20% SBE-CD in saline)1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemESolubility: 2.58 mg/mL (4.49 mM); Clear solution3. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.58 mg/mL (4.49 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 CHIR-99021 (CT99021)個(gè)GSK-3/的抑制劑。IC50 & Target GSK-3 GSK-3 cdc26.7

5、nM (IC50) 10 nM (IC50) 8800 nM (IC50)體外研究 CHIR 99021inhibits human GSK-3 with Ki values of 9.8 nM 1. CHIR 99021 is a small organic molecule thatinhibits GSK3 and GSK3 by competing for their ATP-binding sites.In vitro kinase assays reveal that CHIR99021 specifically inhibits GSK3 (IC50=5 nM) and GSK3

6、 (IC50=10 nM), with little effect on otherkinases 2. In the presence of CHIR-99021 the viability of the ES-D3 cells is reduced by 24.7% at 2.5 M,56.3% at 5 M, 61.9% at 7.5 M and 69.2% at 10 M CHIR-99021 with an IC50 of 4.9 M 3.體內(nèi)研究 In ZDF rats, a single oral dose of CHIR 99021 (16 mg/kg or 48 mg/kg)

7、 rapidly lowers plasma glucose, with amaximal reduction of nearly 150 mg/dl 3-4 h after administration 1. CHIR99021 (2 mg/kg) given once, 4 hbefore irradiation, significantly improves survival after 14.5 Gy abdominal irradiation (ABI). CHIR99021treatment significantly blocks crypt apoptosis and accu

8、mulation of p-H2AX+ cells, and improves cryptregeneration and villus height. CHIR99021 treatment increases Lgr5+ cell survival by blocking apoptosis, andeffectively prevents the reduction of Olfm4, Lgr5 and CD44 as early as 4 h 4.PROTOCOLKinase Assay 2 Kinases are purified from SF9 cells through use

9、 of their His or Glu tag. Glu-tagged proteins are purified, andHis-tagged proteins are purified. Kinase assays are performed in 96-well plates with appropriate peptidesubstrates in a 300-L reaction buffer (variations on 50 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 1 mM EGTA, 1mMdithiothreitol, 25 mM-glycero

10、phosphate, 1 mM NaF, and 0.01% bovine serum albumin). Peptides hasKm values from 1 to 100 M. CHIR 99021 or CHIR GSKIA is added in 3.5 L of Me2SO, followed by ATP toa final concentration of 1 M. After incubation, triplicate 100-L aliquots are transferred to Combiplate 8plates containing 100 L/well of

11、 50 M ATP and 20 mM EDTA. After 1 hour, the wells are rinsed five timeswith phosphate-buffered saline, filled with 200 L of scintillation fluid, sealed, and counted in a scintillationcounter 30 min later. All of the steps are at room temperature. The percentage of inhibition is calculated as100(inhi

12、bitor-no enzyme control)/(Me2SO control-no enzyme control) 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 3 The viability of the mouse ES cells is determined after exposure to different concentrations of GSK3 inhibitorsfor three days using

13、 the MTT assay. The decrease of MTT activity is a reliable metabolism-based test forquantifying cell viability; this decrease correlates with the loss of cell viability. 2,000 cells are seededovernight on gelatine-coated 96-well plates in LIF-containing ES cell medium. On the next day the medium is2

14、/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEchanged to medium devoid of LIF and with reduced serum and supplemented with 0.1-1 M BIO, or 1-10 MSB-216763, CHIR-99021 or CHIR-98014. Basal medium without GSK3 inhibitors or DMSO is used ascontrol. All tested conditions are analyzed in triplicates 3.

15、MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Rats 1Administration 14 Primary hepatocytes from male Sprague Dawley rats that weighed 6cells in 1 mL of DMEM/F12 medium plus0.2% BSA and CHIR 99021(orally at 16 or 48 mg/kg) or controls are incubat

16、ed in 12-well plates on a low-speed shaker for 30 min at 37C in a CO2-enriched atmosphere, collected by centrifugation and lysed byfreeze/thaw in buffer A plus 0.01% NP40; the GS assay is again performed.Mice 4Mice 6-10 weeks old are used. The PUMA+/+ and PUMA-/- littermates on C57BL/6 background (F

17、10) andLgr5-EGFP (Lgr5-EGFP-IRES-creERT2) mice are subjected to whole body irradiation (TBI), or abdominalirradiation (ABI). Mice are injected intraperitoneally (i.p.) with 2 mg/kg of CHIR99021 4 h before radiation or 1mg/kg of SB415286 28 h and 4 h before radiation. Mice are sacrificed to collect s

18、mall intestines for histologyanalysis and western blotting. All mice are injected i.p. with 100 mg/kg of BrdU before sacrifice.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Nat Med. 2016 May;22(5):547-56. Mol Cell. 2017 Mar 2;65(5):873-884

19、.e8. Biomaterials. 2018 Dec 6;193:30-46. Biomaterials. 2018 Oct;180:12-23. Theranostics. 2018 Jul 30;8(15):4262-4278.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Ring DB, et al. Selective glycogen synthase kinase 3 inhibitors potentiate insulin activation of glucose transport and utilization in vitroand in vivo. Diabetes. 2003 Mar;52(3):588-95.2. Bennett CN