NVP-AEW541-AEW541-DataSheet-生命科学试剂-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemENVP-AEW541Cat. No.: HY-50866CAS No.: 475489-16-8Synonyms: AEW541分式: CHNO分量: 439.55作靶点: IGF-1R; Insulin Receptor; Autophagy作通路: Protein Tyrosine Kinase/RTK; Autophagy储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C

2、1 month溶解性数据体外实验 DMSO : 51 mg/mL (116.03 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 2.2751 mL 11.3753 mL 22.7505 mL5 mM 0.4550 mL 2.2751 mL 4.5501 mL10 mM 0.2275 mL 1.1375 mL 2.2751 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。BIOLOGICAL ACTIVITY物活

3、性 NVP-AEW541种有效的 IGF-1R 抑制剂，IC50 为 0.15 M，也抑制 InsR，IC50 为 0.14 M。IC50 & Target IC50: 0.15 0.036 M (IGF-IR), 0.140.039 M (InsR), 0.420.11 M (Flt-3), 20.61 M (PDGFR), 2.40.38M (c-Src), 3.31.4 M (c-Kit) 11/3 Master of Small Molecules 您边的抑制剂师www.MedChemE体外研究 NVP-AEW541 inhibits the in vitro kinase activ

4、ity of the recombinant IGF-IR kinase domain with an IC50 valueof 0.15 M and to be equipotent against the recombinant InsR kinase domain. NVP-AEW541 is confirmedactive toward the IGF-IR kinase (IC50=86 nM) and shown to be selective at the cellular level. Indeed, NVP-AEW541 is found to be 27-fold more

5、 potent toward the native IGF-IR, as compared to the structurally relatednative InsR (IC50=2.3 M). NVP-AEW541 suppresses the IGF-I-mediated survival, soft agar and proliferationof MCF-7 cells with IC50 of 0.162 M, 0.105 M and 1.64 M, respectively 1.体内研究 Oral administration of NVP-AEW541 (20, 30, or

6、50 mg/kg) results in abrogation of basal and IGF-I-inducedreceptor, and PKB and MAPK phosphorylation in the NWT-21 tumor xenograft 1. NVP-AEW541 isadministered by oral gavage 50 mg/kg in 0.2 mL of 25 mM L-(+)-tartaric acid twice a day for 14 consecutivedays. The control group is similarly treated wi

7、th 0.2 mL carrier 25 mM L-(+)-tartaric acid twice a day. Tumorvolume and animal weight are measured thrice a week till the end of the treatment. At that time, animals aresacrificed and tumors are collected and formalin fixed for histologic and immunohistochemical analyses. Inboth cases, NVP-AEW541 t

8、reatment causes tumor shrinkage that reached the statistical significance(P=0.0156 and P=0.0111 for HTLA-230 and SK-N-BE2c, respectively) 2.PROTOCOLKinase Assay 1 The activities of protein kinases are assayed in the presence or absence of inhibitors by measuring theincorporation of 33P from 33PATP (

9、1000 Ci/mmol) into appropriate substrates. The protein kinase assaysare carried out in 96-well plates at RT under conditions described in details below andterminated by theaddition of 20 L of 125 mM EDTA. Subsequently, 30 L (c-Abl, c-Src, IGF-1R) or 40 L (all other kinases) ofthe reaction mixture ar

10、e transferred onto Immobilon-PVDF pre-soaked for 5 min with methanol, rinsed withwater, then soaked for 5 min with 0.5 % H3PO4 and mounted on vacuum manifold. After spotting allsamples, vacuum is connected and each well rinsed with 200 L 0.5 % H3PO4. Membranes are removedand washed 4 on a shaker wit

11、h 1% H3PO4, once with ethanol. After drying, mounting in Packard TopCount96- well frame, and adding of 10 L/well of Microscint, membranes are counted. IC50 values are calculatedby linear regression analysis of the percentage inhibition of each compound in duplicate, at fourconcentrations (usually 0.

12、01, 0.1, 1, and 10 M).One unit of protein kinase activity is defined as 1 nmole of33P transferred from 33PATP to the substrate protein per minute per mg of protein at 37C 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Between 3000 and 60

13、00 cells/well are seeded in 96-well plates with a total media volume of 100 L/well.Increasing concentrations of the compound are added 24 hr thereafter in quadruplicate. 72 hr later, cells arefixed by addition of 25 L/well Glutaraldehyde (20%) and incubation for 10 min at RT. Cells are then washed2

14、with 200L/well H2O and 100L Methylene Blue (0.05%) is added. After incubation for 10 min at RT, cellsare washed 3 with 200 L/well H2O. 200 L/well HCl (3%) is added, and following incubation for 30 min atRT on a plate shaker, absorbance is measured at 650 nm 1.MCE has not independently confirmed the

15、accuracy of these methods. They are for reference only.Animal Mice 1Administration 1 Female Harlan athymic nude mice are used NWT-21 cells are grown in DMEM (high glucose, 4.5 g/L), 10%FCS, 1% L-glutamine, and 1% Na-pyruvate. 5106 cells/animal are initially injected s.c. into the right flank of2/3 M

16、aster of Small Molecules 您边的抑制剂师www.MedChemEfive mice. For the in vivo efficacy experiment, tumors of 500 to 800 mm3 are excised and nonnecrotic areasare cut to fragments of 333 mm. Tumor fragments are washed in sterile PBS and one tumor fragment peranimal is trans- planted s.c. into the right flank

17、. Tumor volumes (lengthwidth height/6) and body weightsare determined three times weekly. At the first day of treatment (day 0), the therapy group (NVP-AEW541)and the control group (vehicle only) are selected by stratification (8 animals per group, average tumor volumeof about 95 mm3 per group). Ani

18、mals are treated p.o. twice daily, 7 days/week either with NVP-AEW541 (20,30, or 50 mg/kg; 10 mL/kg dissolved in 25 mM L(+)-tartaric acid, therapy group) or with 25 mM L(+)-tartaricacid (control group). Antitumor activity is expressed as T/C%(mean increase of tumor volumes of treatedanimals divided

19、by the mean increase of tumor volumes of control animals multiplied by 100). Theexperiment is terminated when the mean tumor volume is about 1500 mm3.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093. Endocr Relat Cancer. 2018 Nov 1. pii: ERC-18-0370.R2. PLoS One. 2018 Feb 7;13(2):e0192214. Harvard Medical School LINCS LIBRARYSee