Nutlin-3a-Rebemadlin-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemENutlin-3aCat.No.:HY-10029CASNo.:675576-98-4Synonyms:Rebemadlin分?式:C??H??Cl?N?O?分?量:581.49作?靶點(diǎn):MDM-2/p53;E1/E2/E3Enzyme;Autophagy;Apoptosis作?通路:Apoptosis;MetabolicEnzyme/Protease;Autophagy儲(chǔ)存?式:Powder-20°C3yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:≥100mg/mL(171.97mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM1.7197mL8.5986mL17.1972mL5mM0.3439mL1.7197mL3.4394mL10mM0.1720mL0.8599mL1.7197mL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(guò)程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過(guò)加熱和/或超聲的?式助溶)1.請(qǐng)依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%saline1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemESolubility:≥2.5mg/mL(4.30mM);Clearsolution2.請(qǐng)依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:2.5mg/mL(4.30mM);Suspendedsolution;Needultrasonic3.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(4.30mM);Clearsolution4.請(qǐng)依序添加每種溶劑：50%PEG300>>50%salineSolubility:8mg/mL(13.76mM);Clearsolution;NeedultrasonicBIOLOGICALACTIVITY?物活性Nutlin-3a(Rebemadlin)Nutlin-3的活性對(duì)映體，?種有效的MDM2抑制劑。Nutlin-3a可抑制MDM2-p53相互作?，穩(wěn)定p53蛋?，從?誘導(dǎo)細(xì)胞?噬(autophagy)和凋亡(apoptosis)。Nutlin-3a有潛??于TP53野?型巢癌的研究。IC50&TargetMDM2-p53[1]體外研究Nutlin-3aisatherapeuticwhichinhibitsMDM2,activateswild-typep53,andinducesapoptosis-asatherapeuticcompoundforTP53wild-typeovariancarcinomas.Threecelllines(HOC-7,OVCA429andA2780)withwild-typeTP53arehighlysensitivetoNutlin-3a(IC50=4to6μM).SKOV3cellshaveanIC50of38μMtoNutlin-3a.Thetworemainingovarianclearcelllines(TOV21GandOVAS),bothwithTP53wild-type,arerelativelymoresensitivetogrowthinhibitionwithNutlin-3a(IC50=14and25μmrespectively)thantheTP53mutantcelllines[1].Nutlin-3aistheactiveenantiomerofNutlin-3.Nutlin-3aisahighlyselectiveMDM2antagonistandp53inducer.Sevendaysofincubationwith10μMNutlin-3aleadsto>90%inhibitionofNIH/3T3cells’growthbutdoesnotaffecttheproliferationofMEFinwhichbothtargetsofthedrugareeliminated.Nutlin-3aeffectivelyarrestescell-cycleprogressioninallcelllines,depletingtheS-phasecompartmentto0.2-2%andincreasingtheG1-andG2/M-phasecompartments,indicatingG1andG2arrest.Thep53targetsp21andMDM2areelevatedsignificantly3hafterNutlin-3aadditionandreachmaximallevelsat8h.Nutlin-3ainducesapoptosisin≈60%ofSJSA-1andMHMcellsafter40h,whichincreasefurtherafter60h(85%and65%,respectively)[2].體內(nèi)研究Nutlin-3aisefficaciousinallmodelswithaveragetumorgrowthinhibition≥98%.Nutlin-3asuppressesxenograftgrowthinadose-dependentfashionwiththehighestdose(200mg/kg)showingasubstantialtumorshrinkage(eightpartialandonefullregressions).TheestablishedSJSA-1andMHMosteosarcomaxenograftswithNutlin-3acausesextensivetumorregression[2].PROTOCOLCellAssay[1]All15celllinesareplatedatadensityof1×103cellsperwellin96-wellplates.After24h,mediaisexchangedandcellsaretreatedwithincrementalconcentrationsofNutlin3a(1μM,5μM,10μM,25μM,50μM,and70μM).After72hofincubation,WST-1isaddedtoeachwell,andamicroplatereaderisusedatanabsorbanceof450nmtomeasurethenumberofremainingviablecells.ExperimentsarerepeatedwithsmallertitrationsofNutlin3aasneededtodeterminetheexactIC50ofcelllines.TheIC50isdefined.Cell2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemElinesareagainplatedinamanneridenticaltoaboveandtreatedwithNutlin3aattheirrespectiveIC50,andWST-1isaddedwithcellviabilitymeasurementat24,48,and72h[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[2]Administration[2]Nudemicebearings.c.tumorxenografts(10micepergroupintheSJSA-1,LnCaP,and22Rv1studyand15micepergroupintheMHMstudy)aredosedorallytwicedailywithNutlin3a(50-200mg/kg)orvehicle(1%Klucel,0.1%Tween80)for2weeks(22Rv1andLnCap)or3weeks(SJSA-1andMHM).Tumorvolumeismeasuredwithacaliperandcalculated.ForWesternblotanalysis,nudemicewithestablishedSJSA-1tumors(200-400mm3,threeanimalspergrouparetreatedwiththreedosesofNutlin3aat150mg/kg(at0,8,and24h),andtumorsareharvested3hafterthelastdose.Tumorsamplesareflash-frozenandprocessed[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶(hù)使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?NatCommun.2022Aug4;13(1):4534.?EMBOJ.2022Jan5;e108946.?EMBOJ.2020Dec22;e104844.?DevCell.2022Feb23;S1534-5807(22)00079-X.?ProcNatlAcadSciUSA.2020Jul28;117(30):17808-17819.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].CraneEK,etal.Nutlin-3a:APotentialTherapeuticOpportunityforTP53Wild-TypeOvarianCarcinomas.PLoSOne.2015Aug6;10(8):e0135101.[2].TovarC,etal.Small-moleculeMDM2antagonistsrevealaberrantp53signalingincancer:implicationsfortherapy.ProcNatlAcadSciUSA.2006Feb7;103(6):1888-93.[3].MUlrich,etal.Murinetumormodelsfortheinvivoevaluationofnaturalc