Pevonedistat-MLN4924-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEPevonedistatCat.No.:HY-70062CASNo.:905579-51-3Synonyms:MLN4924分⼦式:C₂₁H₂₅N₅O₄S分⼦量:443.52作⽤靶点:NEDD8-activatingEnzyme作⽤通路:MetabolicEnzyme/Protease储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:62.5mg/mL(140.92mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.2547mL11.2734mL22.5469mL5mM0.4509mL2.2547mL4.5094mL10mM0.2255mL1.1273mL2.2547mL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.请依序添加每种溶剂：1%DMSO>>99%saline1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemESolubility:0.5mg/mL(1.13mM);Suspendedsolution;Needultrasonic2.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.08mg/mL(4.69mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(4.69mM);Clearsolution4.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(4.69mM);Clearsolution5.请依序添加每种溶剂：10%DMSO>>90%salineSolubility:5mg/mL(11.27mM);Suspendedsolution;Needultrasonic6.请依序添加每种溶剂：5%DMSO>>95%salineSolubility:2.5mg/mL(5.64mM);Suspendedsolution;NeedultrasonicBIOLOGICALACTIVITY⽣物活性Pevonedistat(MLN4924)⼀种有效，选择性的NEDD8活化酶(NAE)抑制剂，IC50为4.7nM。IC50&TargetIC50:4.7nM(NAE)[1]体外研究Pevonedistat(MLN4924)isapotentinhibitorofNAE,andisselectiverelativetothecloselyrelatedenzymesUAE,SAE,UBA6andATG7(IC50=1.5,8.2,1.8and>10μM,respectively)whenevaluatedinpurifiedenzymeassaysthatmonitortheformationofE2-UBLthioesterreactionproducts.Pevonedistat(MLN4924)selectivelyinhibitsNAEactivitycomparedtothecloselyrelatedubiquitin-activatingenzyme(UAE,alsoknownasUBA1)andSUMO-activatingenzyme(SAE;aheterodimerofSAE1andUBA2subunits),inpurifiedenzymeandcellularassays.MLN4924exhibitspotentcytotoxicactivityagainstavarietyofhumantumour-derivedcelllines[1].体内研究Pevonedistat(MLN4924)(sc,10mg/kg,30mg/kg,or60mg/kg)inhibitstheNEDD8pathwayresultinginDNAdamageinMicebearingHCT-116xenografts[1].Pevonedistat(sc,120mg/kg)andTNF-α(10μg/kg)synergisticallycauseliverdamageinSDrats[2].PROTOCOLCellAssay[1]HCT-116cellsgrownin6-wellcell-culturedishesaretreatedwith0.1%DMSO(control)or0.3μMPevonedistat(MLN4924)for24h.Wholecellextractsarepreparedandanalysedbyimmunoblotting.ForanalysisoftheE2-UBLthioesterlevels,lysatesarefractionatedbynon-reducingSDSandimmunoblottedwithpolyclonalantibodiestoUbc12,Ubc9andUbc10.Foranalysisofotherproteins,lysatesarefractionatedbyreducingSDSandprobedwithprimaryantibodiesasfollows:mousemonoclonalantibodiestoCDT1,p27,geminin,ubiquitin,securin/PTTGandp53orrabbitpolyclonalantibodiestoNRF2,CyclinB1andGADD34[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEAdministration[1][2]MicebearingHCT-116tumoursof300-500mm3areadministeredasinglePevonedistat(MLN4924)dose(of10,30or60mg/kg),andtumorsareexcisedatvarioustime-pointsoverthesubsequent24hperiod.TherelativelevelsofNEDD8-cullinandNRF2areestimatedbyquantitativeimmunoblotanalysisusingAlexa680-labelledanti-IgGasthesecondaryantibody.ThestatisticaldifferencebetweenthegroupsforNEDD8-cullininhibitionisdeterminedusingtheKruskal-Wallistest.FortheanalysisofCDT1andphosphorylatedCHK1(Ser317)levelsintumoursections,formalin-fixed,paraffin-embeddedtumoursectionsarestainedwiththerelevantantibodies,amplifiedwithHRP-labelledsecondaryantibodiesanddetectedwiththeChromoMapDABKit.Slidesarecounterstainedwithhaematoxylin.ImagesarecapturedusinganEclipseE800microscopeandRetigaEXicolourdigitalcameraandprocessedusingMetamorphsoftware.CDT1andphosphorylatedCHK1levelsareexpressedasafunctionoftheDABsignalarea.Rats[2]Ten-week-oldmaleSprague-Dawleyratsareused.Acrosstwostudies,atotalofeightanimalsineachgrouparedosedwithvehicle,TNF-α,Pevonedistat(MLN4924),orPevonedistat(MLN4924)+TNF-α.Animalsarefirstintravenouslyadministeredeithervehicle(1×PBS)or10μg/kgTNF-α.Onehourlater,theyaresubcutaneouslyadministeredvehicle(20%sulfobutyletherbeta-cyclodextrinin50mMcitratebuffer,pH3.3)or120mg/kgPevonedistat(MLN4924).Scheduledeuthanasiaoccurred24hpostdose.Unscheduledeuthanasiaisperformedwhenanimalsexhibitedmoribundconditions.SerumiscollectedatnecropsyandanalyzedbyIdexxLaboratoriesforserumchemistrymarkersofliverdamage.Additionally,theliversfromfiveanimalsineachgroupareremoved,separatedintotwosectionsandeitherfrozenat-80°Cforsubsequentproteinanalysisorfixedin10%neutralbufferedformalin,embeddedinparaffin,sectionedat4-6μm,mountedonglassslides,stainedwithhematoxylinandeosin,andanalyzedwithanOlympusBX51lightmicroscopeforhistopathologyassessment.Microscopicfindingsarerecordedinconcordancewiththestandardizednomenclatureforclassifyinglesionswithintheliversofrats.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•Nature.2020Dec;588(7836):164-168.•Cell.2019Jul11;178(2):330-345.e22.•CellRes.2021Mar;31(3):291-311.•CancerCell.2020Mar1