GNE-493 - PI3K 抑制劑 - 生命科學(xué)試劑 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEGNE-493Cat. No.: HY-10811CAS No.: 1033735-94-2分式: CHNOS分量: 372.44作靶點(diǎn): PI3K; mTOR作通路: PI3K/Akt/mTOR儲(chǔ)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(shí)驗(yàn) DMSO : 45 mg/mL (120.82 mM)* means soluble, but sa

2、turation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 2.6850 mL 13.4250 mL 26.8500 mL5 mM 0.5370 mL 2.6850 mL 5.3700 mL10 mM 0.2685 mL 1.3425 mL 2.6850 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。BIOLOGICAL ACTIVITY物活性 GNE-493種有效的，選擇性的 PI3K/mTOR 抑制劑，抑制 PI3K，PI3K，PI3K，PI3K 和 mTOR。IC50

3、 為 3.4 nM，12 nM，16 nM，16 nM 和 32 nM。IC50 & Target PI3K PI3K PI3K PI3K3.4 nM (IC50) 12 nM (IC50) 16 nM (IC50) 16 nM (IC50)1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEmTOR30 nM (IC50)體外研究 GNE-493 is a low molecular weight, potent dual inhibitor of pan-PI3 kinases and mTOR. GNE-493 displaysapproxi

4、mately equipotent inhibition of Class I PI3K isoforms, is submitted for screening in a 142 kinase panelprovided by Invitrogens SelectScreen service. Of these kinases, only three are subject to greater than 50%inhibition by GNE-493, and none are inhibited greater than 80% when tested at 1 M. Subseque

5、ntlymeasured IC50s demonstrated that GNE-493 is more than 100-fold selective for PI3K over these threeunrelated kinases (Aurora A IC5010 M, MLK1 IC50=591 nM and SYK IC50=371 nM) 1.體內(nèi)研究 To confirm and compare in vivo efficacy, GNE-493 is examined in the human MCF7.1 breast cancerxenograft model that

6、harbors a PI3K activating mutation. Mice bearing xenografts are dosed orally oncedaily with 10 mg/kg of GNE-493 for 21 continuous days. Similar to observations made in the PC3 prostatecancer xenograft model, 10 mg/kg of GNE-493 results in 73% tumor growth inhibition at day 21 whencompared to vehicle

7、 control animals. When achieving comparable levels of drug exposure, GNE-493 showsa similar suppression of the PI3K pathway and consequently, a similar efficacy profile against MCF7.1 breasttumors 1.PROTOCOLKinase Assay 1 Enzymatic activity of the Class I PI3K isoforms is measured using a fluorescen

8、ce polarization assay thatmonitors formation of the product 3,4,5-inositoltriphosphate molecule as it competes with fluorescentlylabeled PIP3 for binding to the GRP-1 pleckstrin homology domain protein. An increase in phosphatidylinositide-3-phosphate product results in a decrease in fluorescence po

9、larization signal as the labeledfluorophore is displaced from the GRP-1 protein binding site. Class I PI3K isoforms are expressed andpurified as heterodimeric recombinant proteins. Tetramethylrhodamine-labeled PIP3 (TAMRA-PIP3), di-C8-PIP2, and PIP3 detection reagents are used. PI3K isoforms are ass

10、ayed under initial rate conditions in thepresence of 10 mM Tris (pH 7.5), 25 M ATP, 9.75 M PIP2, 5% glycerol, 4 mM MgCl2, 50 mM NaCl, 0.05%(v/v) Chaps, 1 mM dithiothreitol, 2% (v/v) DMSO at the following concentrations for each isoform: PI3K, PI3 at 60 ng/mL; PI3K at 8 ng/mL; PI3K at 45 ng/mL. After

11、 assay for 30 min at 25C, reactions areterminated with a final concentration of 9 mM EDTA, 4.5 nM TAMRA-PIP3, and 4.2 g/mL GRP-1 detectorprotein before reading fluorescence polarization on an Envision plate reader. IC50s are calculated from the fitof the doseresponse curves to a 4-parameter equation

12、 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 1Administration 1 Human prostate cancer PC3 cells are resuspended in Hanks Balanced Salt Solution and 3106 cellsimplanted subcutaneously into the right hind flank of athymic nu/nu (nude) mic

13、e. Tumors are monitored untilthey reached a mean tumor volume of 150-200 mm3 prior to the initiation of dosing. MCF7.1 cellsresuspended in a 1:1 mixture of Hanks Buffered Salt Solution and Matrigel Basement Membrane Matrix,were 5106 subcutaneously implanted into the right hind flank of athymic nu/nu

14、 (nude) mice. Prior to cellinoculation, 17 -estradiol (0.36 mg/pellet, 60-day release) are implanted into the dorsal shoulder blade areaof each nude mouse. After implantation of cells, tumors are monitored until they reached a mean tumor2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEvolume of 250-

15、350 mm3 prior to initiating dosing. Female nude (nu/nu) mice that are 6-8 weeks old andweighed 20-30 g are used. Tumor bearing mice are dosed orally daily with 10 mg/kg of GNE-493 for 14continuous days. Tumor volume is measured in two dimensions (length and width) and is analyzed usingExcel version

16、11.2. Animal body weights are measured. Tumor sizes are recorded twice weekly over thecourse of the study (14-21 days)MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Sutherlin DP, et al. Discovery of (thienopyrimidin-2-yl)aminopyrimidines as potent, selective, and orally available pan-PI3-kinase anddual pan-PI3-kinase/mTOR inhibitors for the treatment of