ZM-447439-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEZM-447439Cat.No.:HY-10128CASNo.:331771-20-1分?式:C??H??N?O?分?量:513.59作?靶點(diǎn):AuroraKinase;Apoptosis作?通路:CellCycle/DNADamage;Epigenetics;Apoptosis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:25mg/mL(48.68mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM1.9471mL9.7354mL19.4708mL5mM0.3894mL1.9471mL3.8942mL10mM0.1947mL0.9735mL1.9471mL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%salineSolubility:≥2.5mg/mL(4.87mM);Clearsolution1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE2.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(4.87mM);ClearsolutionBIOLOGICALACTIVITY?物活性ZM-447439極光激酶(aurora)抑制劑，對(duì)auroraA和B的IC50值分別為110和130nM。IC50&TargetAuroraAAuroraB110nM(IC50)130nM(IC50)體外研究CellstreatedwithZM-447439progressthroughinterphase,entermitosisnormally,andassemblebipolarspindles.However,chromosomealignment,segregation,andcytokinesisallfail.ZM-447439inhibitscelldivisionandinhibitmitoticphosphorylationofhistoneH3.ZM-447439preventschromosomealignmentandsegregation.ZM-447439compromisesspindlecheckpointfunction.ZM-447439inhibitskinetochorelocalizationofBubR1,Mad2,andCenp-E[1].InhibitionofAurorakinasebyZM-447439reduceshistoneH3phosphorylationatSer10inHep2carcinomacells.MultipolarspindlesareinducedintheseZM-treatedG2/M-arrestedcellswithaccumulationof4N/8NDNA,similartocellswithgeneticallysuppressedAurora-B.ZM-447439treatmentinducescellapoptosis.ZM-447439inhibitionofAurorakinaseispotentlyinassociationwithdecreaseofAktphosphorylationatSer473anditssubstratesGSK3α/βphosphorylationatSer21andSer9[2].PROTOCOLKinaseAssay[1]1ngpurifiedrecombinantenzymeisaddedtoareactioncocktailcontainingbuffer,10μMpeptidesubstrate,10μMforAuroraAor5μMATPforAuroraB,and0.2μCiγ[33P]ATP,andisthenincubatedatroomtemperaturefor60min.Reactionsarestoppedbyadditionof20%phosphoricacid,andtheproductsarecapturedonP30nitrocellulosefiltersandassayedforincorporationof33PwithaBetaplatecounter.NoenzymeandnocompoundcontrolvaluesareusedtodeterminetheconcentrationofZM-447439,whichgave50%inhibitionofenzymeactivity[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]Todeterminecloningefficiency,MCF7cellsareplatedinphenolredfreeDMEplus5%strippedserum,andarethentreatedwithorwithouttheanti-estrogenICI182780at1μMfor48h.ZM-447439isthenaddedattheindicatedconcentrationsfor72h.Thecellsareharvested,washed,and-400cellsplatedineachwellofa6-wellplateincompletemediawithoutZM-447439.After10d,thecoloniesarefixed,stainedwithcrystalviolet,andcounted.ThecloningefficiencyrepresentsthenumberofcoloniesonZM-447439-treatedplatescomparedwithDMSO-treatedcontrols[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?EBioMedicine.2021Aug5;70:103510.2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE?Elife.2020Dec7;9:e61405.?bioRxiv.2021Feb5.?HarvardMedicalSchoolLINCSLIBRARYSeemorecustomervalidationsonwww.MedChemEREFERENCES[1].DitchfieldC,etal.AuroraBcoupleschromosomealignmentwithanaphasebytargetingBubR1,Mad2,andCenp-Etokinetochores.JCellBiol.2003Apr28;161(2):267-80.[2].LongZJ,etal.ZM447439inhibitionofaurorakinaseinducesHep2cancercellapoptosisinthree-dimensionalculture.CellCycle.2008May15;7(10):1473-9.MceP