Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEVE-821Cat.No.:HY-14731CASNo.:1232410-49-9分?式:C??H??N?O?S分?量:368.41作?靶點:ATM/ATR作?通路:CellCycle/DNADamage;PI3K/Akt/mTOR儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數據體外實驗DMSO:50mg/mL(135.72mM;Needultrasonic)H2O:<0.1mg/mL(ultrasonic;warming;heatto60°C)(insoluble)MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.7144mL13.5718mL27.1437mL5mM0.5429mL2.7144mL5.4287mL10mM0.2714mL1.3572mL2.7144mL請根據產品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；?旦配成溶液，請分裝保存，避免反復凍融造成的產品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲存時，請在6個?內使?，-20°C儲存時，請在1個?內使?。體內實驗VE-821ispreparedimvehicle(10%PEG300,2.5%Tween-80,pH4)[4].BIOLOGICALACTIVITY?物活性VE-821?種有效的ATP競爭性的ATR抑制劑，Ki/IC50為13nM/26nM。1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEIC50&TargetATRATMDNA-PKPI3Kγ13nM(Ki)16μM(Ki)2.2μM(Ki)3.9μM(Ki)體外研究VE-821showsexcellentselectivityforATRwithminimalcross-reactivityagainsttherelatedPIKKsATM,DNA-PK,mTORandPI3Kγ(Kisof16μM,2.2μM,>1μMand3.9μM,respectively)andagainstalargepanelofunrelatedproteinkinases[1].VE-821(compound27)alsoinhibitsATMandDNA-PKwirhIC50of>8μM,and4.4μM,respectively[2].VE-821significantlyenhancesthesensitivityofPSN-1,MiaPaCa-2andprimaryPancMpancreaticcancercellstoradiationandGemcitabineunderbothnormoxicandhypoxicconditions.ATRinhibitionbyVE-821leadstoinhibitionofradiation-inducedG2/Marrestincancercells.InbothPSN-1andMiaPaCa-2cells,1μMVE-821inhibitsphosphorylationofChk1(Ser345)aftertreatmentwithGemcitabine(100nM),radiation(6Gy)orboth,at2hpost-irradiation[3].PROTOCOLKinaseAssay[2]Theabilityofcompounds(e.g.,VE-821)toinhibitATR,ATMorDNAPKkinaseactivityistestedusingaradiometric-phosphateincorporationassay.Astocksolutionispreparedconsistingoftheappropriatebuffer,kinase,andtargetpeptide.Tothisisaddedthecompoundofinterest,atvaryingconcentrationsinDMSOtoafinalDMSOconcentrationof7%.Assaysareinitiatedbyadditionofanappropriate[g-33P]ATPsolutionandincubatedat25°C.Assaysarestopped,afterthedesiredtimecourse,byadditionofphosphoricacidandATPtoafinalconcentrationof100mMand0.66μM,respectively.Peptidesarecapturedonaphosphocellulosemembrane,prepared,andwashedsixtimeswith200μLof100mMphosphoricacid,priortotheadditionof100μLofscintillationcocktailandscintillationcountingona1450MicrobetaLiquidScintillationCounter.Dose?responsedataareanalyzedusingGraphPadPrismsoftware[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[3]MiaPaCa-2,PSN-1andPanc1cells(5×104)areplatedin96-wellplatesandafter4htreatedwithincreasingconcentrationsofVE-821at1hbeforeirradiationwithasingledoseof4Gy.Mediumisreplaced72hpost-irradiationatwhichpointviabilityismeasuredusingtheusingtheAlamarBlueassay.Cellsareallowedtoproliferateandcellviabilityisagainanalyzedatday10forthedifferenttreatmentconditions.Cellviabilityandsurvivingfractionarenormalizedtotheuntreated(control)group[3].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產品發表的科研?獻?MolCell.2022Apr14:S1097-2765(22)00290-8.?SciTranslMed.2018Jul18;10(450).pii:eaaq1093.?NucleicAcidsRes.2020Sep18;48(16):9109-9123.?NatCommun.2018Oct8;9(1):4139.?CellSyst.2020Jan22;10(1):66-81.e11.Seemorecustomervalidationsonwww.MedChemEREFERENCES2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE[1].ReaperPM,etal.SelectivekillingofATM-orp53-deficientcancercellsthroughinhibitionofATR.NatChemBiol.2011Apr13;7(7):428-30.[2].CharrierJD,etal.DiscoveryofpotentandselectiveinhibitorsofataxiatelangiectasiamutatedandRad3related(ATR)proteinkinaseaspotentialanticanceragents.JMedChem.2011Apr14;54(7):2320-30.[3].PrevoR,etal.ThenovelATRinhibitorVE-821increasessensitivityofpancreaticcancercellstoradiationandchemotherapy.CancerBiolTher.2012Sep;13(11):1072-81.[4].MuralidharanSV,etal.BETbromodomaininhibitorssynergizewithATRinhibitorstoinduceDNAdamage,apoptosis,senescence-associatedsecretorypathwayandERstressinMyc-inducedlymphomacells.Oncogene.2016Sep