Pinometostat-EPZ-5676-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEPinometostatCat.No.:HY-15593CASNo.:1380288-87-8Synonyms:EPZ-5676分⼦式:C₃₀H₄₂N₈O₃分⼦量:562.71作⽤靶点:HistoneMethyltransferase作⽤通路:Epigenetics储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:100mg/mL(177.71mM;Needultrasonic)Ethanol:100mg/mL(177.71mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM1.7771mL8.8856mL17.7711mL5mM0.3554mL1.7771mL3.5542mL10mM0.1777mL0.8886mL1.7771mL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；⼀旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存⽅式和期限：-80°C,6months;-20°C,1month。-80°C储存时，请在6个⽉内使⽤，-20°C储存时，请在1个⽉内使⽤。体内实验请根据您的实验动物和给药⽅式选择适当的溶解⽅案。以下溶解⽅案都请先按照InVitro⽅式配制澄的储备液，再依次添加助溶剂：(为保证实验结果的可靠性，澄的储备液可以根据储存条件，适当保存；体内实验的⼯作液，建议您现⽤现配，当天使⽤；以下溶剂前显⽰的百分⽐指该溶剂在您配制终溶液中的体积占⽐；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的⽅式助溶)1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE1.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(4.44mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(4.44mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(4.44mM);Clearsolution4.请依序添加每种溶剂：10%EtOH>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(4.44mM);Clearsolution5.请依序添加每种溶剂：10%EtOH>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(4.44mM);Clearsolution6.请依序添加每种溶剂：10%EtOH>>90%cornoilSolubility:≥2.5mg/mL(4.44mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性Pinometostat(EPZ-5676)有效的DOT1L组蛋⽩甲转移酶抑制剂，Ki为80pM。IC50&TargetKi:<80pM(DOT1Lhistonemethyltransferase)体外研究Pinometostat(EPZ-5676)inhibitsH3K79me2withIC50valuesof3nMand5nMinMV4-11andHL60cells,respectively.Pinometostat(EPZ-5676)isapotentinhibitorofMV4-11proliferationwithanIC50valueof3.5nM[1].Pinometostat(EPZ-5676)inducesasynergisticanddurableantiproliferativeeffect,increasesexpressionofdifferentiationmarkersandapoptosisassingleagent,anddemonstratescombinationbenefitincombinationwithAMLstandardofcaredrugsinMLL-rcells[2].体内研究Pinometostat(EPZ-5676)(70mg/kg,i.p.)causescompleteandsustainedregressioninaratxenograftmodelofMLL-rearrangedleukemia.Pinometostat(EPZ-5676)(70,35mg/kg,i.v.)reducesHOXA9andMEIS1mRNAlevelsoftumorstakenfromrats,andreducesMLL-fusiontargetgeneexpressioninvivo[1].PROTOCOLCellAssay[1]ToanalyseinhibitionofhistonemethylationinMV4-11cellsfollowingPinometostattreatment,extractedhistones(400ng)arefractionatedona10-20%TrisHClgelswithTris-GlycineSDSrunningbufferunderdenaturingconditionsandtransferredtonitrocellulosefilters.Filtersarecutintostripsandincubatedfor1hourinblockingbufferatroomtemperature(RT)andthenincubatedovernightat4°Cinblockingbuffer.Filtersarewashed3timesfor5minuteswithwashbuffer(Phosphatebufferedsaline(PBS)including0.01%Tween20(PBST))andincubatedwithinfraredtaggedsecondaryantibodyatRTfor1hour.FiltersarewashedinPBSTandreprobedfor1houratRTwiththeappropriatetotalhistoneantibodycontrol(mouseanti-histoneH3(1:20,000),CST3638,ormouseanti-histoneH4(1:10,000),CST2935).FiltersarewashedagaininPBSTandincubatedwithinfraredtaggedsecondaryantibody(IRDye800Cwdonkey-anti-mouseIgG(1:20,000),Li-Cor926-32212)atRTfor1hour.AfterafinalishinPBST,filtersarescannedusingtheOdysseyinfaredimager(Li-cor).ToanalyseinhibitionofH3K79methylationinperipheralbloodmononuclear2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEcells(PBMCs)fromratsdosedwithPinometostat(EPZ-5676),20μLofPBMCwholecelllysateisfractionatedondenaturinggelsandanalysedbyimmunoblottingwithantibodiestoH3K79me2ortotalH3.SignalintensitiesspecificfortheH3K79me2antibodyandtotalhistoneH3controlantibodyarequantifiedusingOdysseysoftware.TheH3K79me2signalintensityisnormalizedbydividingitbythetotalhistoneH3controlsignalintensityinthesamelane.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.Animal0.2mLofaMV4-11cellsuspension(1×107cells)inPBSisinjectedsubcutaneouslyintofemaleathymicAdministration[1]nudemice(Crl:NU(Ncr)-Foxn1nu).Tumorsaremeasuredbycalipersandmicearerandomizedaccordingtotumorsizeintotreatmentgroups(n=10)beforetheinitiationofdosingwithPinometostat(EPZ-5676)whentumorvolumesreacheapproximately100mma3.Pinometostatisadministeredintraperitoneallythreetimesdailyfor28daysat10and20mg/kgin10%ethanolinsaline.Miceareweighedandtumorsmeasuredwithcaliperstwiceweeklyuntiltheendofthestudy.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•CellSyst.2018Apr25;6(4):424-443.e7.•CellRep.2021Sep21;36(12):109739.•CellDeathDis.2018Jan26;9(2):129.•ActaPharmacolSin.2021Apr13.•Phytomedicine.2021,153499.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].DaigleSR,etal.PotentinhibitionofDOT1LastreatmentforMLL-fusionleukemia.Blood.2013Jun25.[Epubaheadofprint][2].KlausCR,etal.DOT1LinhibitorEPZ-5676displayssynergisticantiproliferativeactivityincombinationwithstandardofcaredrugsandhypomethylatingagentsinMLL-rear