NMS-873 - p97 抑制剂 - 生命科学试剂 - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemENMS-873Cat. No.: HY-15713CAS No.: 1418013-75-8分式: CHNOS分量: 520.67作靶点: p97作通路: Cell Cycle/DNA Damage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 : 20.5 mg/mL (39.37 mM; Need ultrasonic and

2、warming)Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 1.9206 mL 9.6030 mL 19.2060 mL5 mM 0.3841 mL 1.9206 mL 3.8412 mL10 mM 0.1921 mL 0.9603 mL 1.9206 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物活性 NMS-873种有效，选择性的 VCP/p97 变构抑制剂，IC50 值为 30 nM。IC50 & Target IC50: 30 nM 1体外研究

3、NMS-873 has antiproliferative effect on a panel of tumor cell lines with IC50 values in the range of 0.08 M to2 M. For HCT116 and HeLa cells, the IC50 values are 0.4 M and 0.7 M, respectively. NMS-873 reducesVCP sensitivity to trypsin digestion, preventing degradation of the linker-D2 domain. NMS-87

4、3 induces clear,1/2 Master of Small Molecules 您边的抑制剂师www.MedChemEdose-dependent accumulation of poly-Ub proteins and stabilization of cyclin E and Mcl-1 at doses consistentwith its antiproliferative IC50 value 1.PROTOCOLKinase Assay 1 The ATPase activity and the kinetic parameters of recombinant wil

5、d-type VCP and its mutants are evaluatedby monitoring ADP formation in the reaction, using a modified NADH-coupled assay46.As ADP and NADH are ATP-competitive inhibitors of VCP ATPase activity, the standard protocol for theNADH-coupled assay is modified into a two-step procedure. In the first part,

6、an ATP-regenerating system (40U/mL pyruvate kinase and 3 mM phosphoenolpyruvate) recycles the ADP produced by VCP activity, keepsthe substrate concentration constant (thus preventing product inhibition) and accumulates a stoichiometricamount of pyruvate. In the second part, the VCP enzymatic reactio

7、n is quenched with 30 mM EDTA and 250M NADH and stoichiometrically oxidized by 40 U/mL lactic dehydrogenase to reduce accumulated pyruvate.The decrease of NADH concentration is measured at 340 nm using a Tecan Safire 2 reader plate. The assayis performed in 96- or 384-well UV platesin a reaction buf

8、fer with 50 mM Hepes, pH 7.5, 0.2 mg/mL BSA, 10mM MgCl2 and 2 mM DTT.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Cells are seeded at 1,600 cells per well in 384-well white clear-bottom plates. Twenty-four hours afterseeding, cells are t

9、reated with the compounds (eight dilution points, in duplicate, for each compound) andincubated for an additional 72 h at 37C under a 5% CO2 atmosphere. Cells are then lysed, andthe ATP content in each well is determined using a thermostable firefly luciferase-based assay from Promegaas a measure of

10、 cell viability. IC50 values are calculated using the percentage of growth of treated cellsversus the untreated control.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 Cell Chem Biol. 2019 Jun 4. pii: S2451-9456(19)30175-8. J Cell Mol Med. 2016 Jan;20(1):58-70.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Paola Magnaghi, et al. Covalent and allosteric inhibitors of the ATPATP ase VCP/p97 induce cancer cell death. Nat Chem Biol. 2013 Jul28. doi: 10.1038/nchembio.1313.McePdfHeightCaution: Product has not be