GSK-690693 - Akt 抑制剂 - 生命科学试剂 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEGSK-690693Cat. No.: HY-10249CAS No.: 937174-76-0分式: CHNO分量: 425.48作靶点: Akt; Autophagy; AMPK作通路: PI3K/Akt/mTOR; Autophagy; Epigenetics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 : 25 mg/mL

2、 (58.76 mM; Need ultrasonic)Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 2.3503 mL 11.7514 mL 23.5029 mL5 mM 0.4701 mL 2.3503 mL 4.7006 mL10 mM 0.2350 mL 1.1751 mL 2.3503 mL请根据产品在不同溶剂中的溶解度，选择合适的溶剂配制储备液，并请注意储备液的保存式和期限。体内实验 请根据您的实验动物和给药式选择适当的溶解案，配制前请先配制澄清的储备液，再依次添加助溶剂(为保证实验结果的可靠性，体内实验的作液，建议您现现配，

3、当天使；澄清的储备液可以根据储存条件，适当保存；以下溶剂前的百分 指该溶剂在您配制终溶液中的体积占)：1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (5.88 mM); Clear solution2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (5.88 mM); Clear solution3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5

4、 mg/mL (5.88 mM); Clear solution1/4 Master of Small Molecules 您边的抑制剂师www.MedChemEBIOLOGICAL ACTIVITY物活性 GSK-690693是ATP竞争型的 Akt 抑制剂，对Akt1，Akt2和Akt3 的IC50 分别为2，13，9 nM。IC50 & Target Akt1 Akt3 Akt2 PKC2 nM (IC50) 9 nM (IC50) 13 nM (IC50) 2 nM (IC50)PKC PrkX PAK6 PAK42 nM (IC50) 5 nM (IC50) 6 nM (IC50)

5、10 nM (IC50)PKC PKC1 PKC PKA14 nM (IC50) 19 nM (IC50) 21 nM (IC50) 24 nM (IC50)PKG1 AMPK PAK5 DAPK333 nM (IC50) 50 nM (IC50) 52 nM (IC50) 81 nM (IC50)Autophagy体外研究 GSK690693 is very selective for the Akt isoforms versus the majority of kinases in other families. However,GSK690693 is less selective f

6、or members of the AGC kinase family including PKA, PrkX, and PKC isozymeswith IC50 of 24 nM, 5 nM, and 2-21 nM, respectively. GSK690693 also potently inhibits AMPK and DAPK3from the CAMK family with IC50 of 50 nM and 81 nM, respectively, and PAK4, 5, and 6 from the STE familywith IC50 of 10 nM, 52 n

7、M, and 6 nM, respectively. GSK690693 inhibits the phosphorylation of GSK3 intumor cells with IC50 ranging from 43 nM to 150 nM. GSK690693 treatment leads to a dose-dependentincrease in the nuclear accumulation of the transcription factor FOXO3A. GSK690693 potently inhibits theproliferation of T47D,

8、ZR-75-1, BT474, HCC1954, MDA-MB-453, and LNCaP cells with IC50 of 72 nM, 79nM, 86 nM, 119 nM, 975 nM, and 147 nM, respectively. GSK690693 treatment induces apoptosis atconcentrations 100 nM in both LNCaP and BT474 cells 1. Consistent with the role of AKT in cell survival,GSK690693 induces apoptosis

9、in sensitive ALL cell lines 2.体内研究 A single administration of GSK690693 inhibits GSK3 phosphorylation in human breast carcinoma (BT474)xenografts in a dose- and time-dependent manner. Similarly, GSK690693 induces a reduction inphosphorylation of the Akt substrates, PRAS40, and FKHR/FKHRL1. GSK690693

10、 also results in an acuteincrease in blood glucose, returning to baseline 8 to 10 hours after drug administration. Administration ofGSK690693 induces reductions in phosphorylated Akt substrates in vivo, and potently inhibits the growth ofhuman SKOV-3 ovarian, LNCaP prostate, and BT474 and HCC-1954 b

11、reast carcinoma xenografts, withmaximal inhibition of 58% to 75% at the dose of 30 mg/kg/day 1. GSK690693 exhibits efficacy irrespectiveof the mechanism of Akt activation involved. GSK690693 is most effective in delaying tumor progression inLck-MyrAkt2 mice expressing a membrane-bound, constitutivel

12、y active form of Akt 3.PROTOCOLKinase Assay 1 His-tagged full-length Akt1, 2, or 3 are expressed and purified from baculovirus. Activation is carried out with2/4 Master of Small Molecules 您边的抑制剂师www.MedChemEpurified PDK1 to phosphorylate Thr308 and purified MK2 to phosphorylate Ser473. To more accur

13、atelymeasure time-dependent inhibition of Akt, activated Akt enzymes are incubated with GSK690693 at variousconcentrations at room temperature for 30 minutes before the reaction is initiated with the addition ofsubstrate. Final reaction contains 5 nM to 15 nM Akt1, 2, and 3 enzymes; 2 M ATP; 0.15 Ci

14、/L-33PATP;1 M Peptide (Biotin-aminohexanoicacid-ARKR-ERAYSFGHHA-amide); 10 mM MgCl2; 25 mM MOPS (pH7.5); 1 mM DTT; 1 mM CHAPS; and 50 mM KCl. The reactions are incubated at room temperature for 45minutes, followed by termination with Leadseeker beads in PBS containing EDTA (final concentration, 2mg/

15、mL beads and 75 mM EDTA). The plates are then sealed, the beads are allowed to settle for at least 5hours, and product formation is quantitated using a Viewlux Imager.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Cells are plated at densi

16、ties that allow untreated cells to grow logarithmically during the course of a 3-dayassay. Briefly, cells are plated in 96- or 384-well plates and incubated overnight. Cells are then treated withGSK690693 (ranging from 30 M-1.5 nM) and incubated for 72 hours. Cell proliferation is measured usingthe

17、CellTiter Glo reagent. Data are analyzed using the XLFit curve-fitting tool for Microsoft Excel. IC50 valuesare obtained by fitting data to Eq, 2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Tumors are initiated by injection of tumor cell susp

18、ension (HCC1954, MDA-MB-453, and LNCaP) or tumorAdministration 1 fragments (BT474, SKOV-3, and PANC1) s.c. in 8- to 12-wk-old CD1 Swiss Nude mice (LNCaP, SKOV-3,and PANC1) or SCID mice (HCC1954, MDA-MB-453, and BT474). When tumors reach a volume of 100 to200 mm3, mice are randomized and divided into

19、 groups of 8 to 12 mice per group. GSK690693 isadministered once daily at 10, 20, and 30 mg/kg by i.p. administration. Animals are euthanized by inhalationof CO2 at the completion of the study. Tumor volume is measured twice weekly by calipers, using theequation: tumor volume (mm3)=(length width2)/2

20、. Results are reported as % inhibition on day 21 oftreatment=100 1-(average growth of the drug-treated population/average growth of vehicle-treated controlpopulation). Statistical analysis is done using two-tailed t test.MCE has not independently confirmed the accuracy of these methods. They are for

21、 reference only.户使本产品发表的科研献 Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093. Oxid Med Cell Longev. 2019 Apr 28;2019:2717986. PLoS One. 2019 May 15;14(5):e0216759. PLoS One. 2016 Jan 28;11(1):e0147682. Oncotarget. 2016 May 17;7(20):29131-42.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Rhodes N, et al. Characterizat