Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEPevonedistatCat.No.:HY-70062CASNo.:905579-51-3Synonyms:MLN4924分?式:C??H??N?O?S分?量:443.52作?靶點(diǎn):NEDD8-activatingEnzyme作?通路:MetabolicEnzyme/Protease儲(chǔ)存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:62.5mg/mL(140.92mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM2.2547mL11.2734mL22.5469mL5mM0.4509mL2.2547mL4.5094mL10mM0.2255mL1.1273mL2.2547mL請(qǐng)根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲(chǔ)備液；?旦配成溶液，請(qǐng)分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲(chǔ)備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?，-20°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?。體內(nèi)實(shí)驗(yàn)請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥?式選擇適當(dāng)?shù)娜芙?案。以下溶解?案都請(qǐng)先按照InVitro?式配制澄的儲(chǔ)備液，再依次添加助溶劑：(為保證實(shí)驗(yàn)結(jié)果的可靠性，澄的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)保存；體內(nèi)實(shí)驗(yàn)的?作液，建議您現(xiàn)?現(xiàn)配，當(dāng)天使?；以下溶劑前顯?的百分?指該溶劑在您配制終溶液中的體積占?；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的?式助溶)1.請(qǐng)依序添加每種溶劑：1%DMSO>>99%saline1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemESolubility:0.5mg/mL(1.13mM);Suspendedsolution;Needultrasonic2.請(qǐng)依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.08mg/mL(4.69mM);Clearsolution3.請(qǐng)依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(4.69mM);Clearsolution4.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(4.69mM);Clearsolution5.請(qǐng)依序添加每種溶劑：10%DMSO>>90%salineSolubility:5mg/mL(11.27mM);Suspendedsolution;Needultrasonic6.請(qǐng)依序添加每種溶劑：5%DMSO>>95%salineSolubility:2.5mg/mL(5.64mM);Suspendedsolution;NeedultrasonicBIOLOGICALACTIVITY?物活性Pevonedistat(MLN4924)?種有效，選擇性的NEDD8活化酶(NAE)抑制劑，IC50為4.7nM。IC50&TargetIC50:4.7nM(NAE)[1]體外研究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].體內(nèi)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?邊的抑制劑?師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.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?Nature.2020Dec;588(7836):164-168.?Cell.2019Jul11;178(2):330-345.e22.?CellRes.2021Mar;31(3):291-311.?CancerCell.2020Mar1