PHT-427 - Akt 抑制劑 - 生命科學(xué)試劑 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEPHT-427Cat. No.: HY-12063CAS No.: 1191951-57-1分式: CHNOS分量: 409.61作靶點: Akt作通路: PI3K/Akt/mTOR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 : 50 mg/mL (122.07 mM; Need ultrasonic)Mass Solvent1

2、 mg 5 mg 10 mg Concentration制備儲備液1 mM 2.4413 mL 12.2067 mL 24.4135 mL5 mM 0.4883 mL 2.4413 mL 4.8827 mL10 mM 0.2441 mL 1.2207 mL 2.4413 mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存式和期限。體內(nèi)實驗 請根據(jù)您的實驗動物和給藥式選擇適當(dāng)?shù)娜芙獍福渲魄罢埾扰渲瞥蔚膬湟?，再依次添加助溶?為保證實驗結(jié)果的可靠性，體內(nèi)實驗的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使；澄的儲備液可以根據(jù)儲存條件，適當(dāng)保存；以下溶劑前的百分 指該溶劑在您配制終溶

3、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: 2.5 mg/mL (6.10 mM); Clear solution2. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (6.10 mM); Clear solutionBIOLOGICAL ACTIVITY1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE物活性 PHT-427種 Akt 和 PDPK1 雙重 抑制劑，與 Akt 和

4、 PDPK1 中的 PH 結(jié)構(gòu)域有親和性，Ki 分別為 2.7 M 和5.2 M。IC50 & Target Akt PDPK12.7 M (Ki) 5.2 M (Ki)體外研究 The effects of PHT-427 on cell signaling are investigated by RPPA using a panel of 86 antibodies tophospho- and non-phosphorylated signaling protein related to PtdIns-3-K/PDPK1/Akt signaling in PC-3prostate cel

5、ls where PtdIns-3-K/PDPK1/Akt signaling is activated because of homozygous PTEN mutation.After 16 hours, a reduction is observed in phospho-Ser241-PDPK1 phospho-Thr308-Akt by both 10 M PH-427 and 0.1 M Wortmannin. Finally, phospho-Ser657-protein kinase C (PKC) and total SGK1 are decreasedby treatmen

6、t with both PHT-427 and Wortmannin. These results suggest that at 10 M PHT-427 inhibits bothAkt and PDKP1. The BxPC-3 and MiaPaCa-2 pancreatic cancer cell lines are probed by Western blottingfollowing up to 24 hr exposure to 10 M PHT-427, which is below the IC50 for cell growth inhibition of around3

7、0 M, to determine the effects of PHT-427 on of the PtdIns-3-K/PDPK1/Akt signaling pathway components1.體內(nèi)研究 Mice with BxPC-3 pancreatic, MCF-7 breast or A-549 NSCL cancer xenografts are administered PHT-427, orits analogs with a C-4, C-6 or C-8 alkyl chain by oral gavage twice a day for 10 days. The

8、results show thatPHT-427 has the greatest antitumor activity with the C-8 chain analog having less activity, and analogs with aC-4 or C-6 chain very little activity. All further antitumor studies are conducted using compound PHT-427.Plasma levels of PHT-427 following oral administration to mice of a

9、 dose of 200 mg/kg show rapidabsorption, without a lag phase, Cmax is 8.2 g/mL 1 hr following dosing, and the elimination half-life is 1.4hr with a terminal PHT-427 concentration of 0.1 g/mL 10 hr after dosing. The plasma concentration of PH-427 is above the level which gave inhibition of Akt and PD

10、PK1 signaling in cells of 10 M (4 g/mL) for atleast 3 hr 1.PROTOCOLCell Assay 1 Panc-1 cells stably transfected with green fluorescent protein (GFP) tagged Akt or PDKP1 PH domains areserum starved in phenol red free growth medium on glass-bottom 96-well imaging plates for 16 hours. Theyare then trea

11、ted with PHT-427 at 1, 5, and 10 M or PI-103 for 4 hr, and stimulated with 50 ng/mL IGF-1 for10 min. Images are taken before and after IGF-1 treatment using an IN Cell Analyzer 1000 instrument with aNikon Plan Fluor ELWD 20X/0.45 objective loaded and using a 300msec exposure time 1.MCE has not indep

12、endently confirmed the accuracy of these methods. They are for reference only.Animal Mice 1Administration 1 Female C57Bl/6 mice are administered PHT-427 as a single oral dose of 200 mg/kg. The mice are killed atdifferent times (3 mice at each time point), blood collected into heparinized tubes and p

13、lasma prepared andstored frozen at -80C. For assay 0.2 mL plasma is mixed with 0.2 mL of 0.1 M sodium phosphate buffer, pH4.0, and extracted for 1 hr by inversion with 1 ml ethyl acetate. After centrifugation 0.8 mL of the organic layeris removed, evaporated under N2 and redissolved in 0.2 mL ethano

14、l and 10 L injected onto a WatersQuattro Ultima tandem mass spectrometer using a Phenomenex Luna 3.0 m, 2.050 mm C8 analytical2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEcolumn with detection and quantification by multiple reaction monitoring with the mass spectrometeroperating in electrospray

15、positive ionization mode.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 J Funct Foods. 2019 Feb.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Meuillet EJ, et al. Molecular pharmacology and antitumor activity of PHT-427, a novel Akt/phosphatidylinositide-dependent proteinkinase 1 pleckstrin homology domain inhibitor. Mol Canc