Tozasertib-VX-680-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemETozasertibCat.No.:HY-10161CASNo.:639089-54-6Synonyms:VX680;MK-0457分⼦式:C₂₃H₂₈N₈OS分⼦量:464.59作⽤靶点:AuroraKinase;Autophagy作⽤通路:CellCycle/DNADamage;Epigenetics;Autophagy储存⽅式:4°C,protectfromlight*Insolvent:-80°C,6months;-20°C,1month(protectfrom

light)溶解性数据体外实验DMSO:≥106.67mg/mL(229.60mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制备储备液1mM2.1524mL10.7622mL21.5244mL5mM0.4305mL2.1524mL4.3049mL10mM0.2152mL1.0762mL2.1524mL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(protectfromlight)。-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：10%DMSO>>40%PEG300>>5%Tween-80>>45%saline1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemESolubility:≥2.08mg/mL(4.48mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(4.48mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(4.48mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性Tozasertib(VX680;MK-0457)AuroraA/B/C激酶抑制剂，Ki值分别为0.6，18，4.6nM。IC50&TargetAuroraAAuroraBAuroraC0.6nM(Ki)18nM(Ki)4.6nM(Ki)体外研究TozasertibinducessimilarcytotoxicitywithIC50ofapproximately300nMandexhibitsanAURB-likeinhibitoryphenotypeofG2/Marrest,endoreduplicationandapoptosisinBaF3cellstransfectedwithABLorFLT-3(mutantandwildtype)kinases.TozasertibpreventstheCAL-62proliferationinatime-dependentmanner.Tozasertibtreatmentfor14dayssignificantlydecreasesthenumberandsizeofcoloniesbyapproximately70%inthe8305Cand90%intheCAL-62,8505CandBHT-101.TreatmentofthedifferentATCcellswithTozasertibinhibitsproliferationwiththeIC50between25and150nM.TheTozasertibsignificantlyimpairstheabilityofthedifferentcelllinestoformcoloniesinsoftagar.Analysisofcaspase-3activityindicatesthatTozasertibinducesapoptosisinthedifferentcelllines.CAL-62cellsexposedfor12hourstoTozasertibshowanaccumulationofcellswith≥4NDNAcontent.Time-lapseanalysisdemonstratesthatTozasertib-treatedCAL-62cellsexitmetaphasewithoutdividing.Moreover,histoneH3phosphorylationisabrogatedfollowingTozasertibtreatment[2].TozasertibhassignificantinhibitoryactivityagainstBCR-AblbearingtheT315Imutationinpatient-derivedsamples[3].PROTOCOLKinaseAssay[3]TheconsumptionofATPiscoupledviathepyruvatekinase/lacticdehydrogenaseenzymepairtotheoxidationofNADH,whichcanbemonitoredthroughthedecreaseinabsorptionat340nm.Reactionscontains100mMTris(pH8),10mMMgCl2,2.2mMATP,1mMphosphoenolpyruvate,0.6mg/mLNADH,75units/mLpyruvatekinase,105units/mLlactatedehydrogenase,and0.5mMsubstratepeptide(sequence:EAIYAAPFAKKK).Reactions(75μL)arestartedbyaddingsufficientkinasetobringthereactionsto30nMkinaseconcentrationandthedecreaseinabsorbanceismonitoredover30minutesat30°Cinamicrotiterplatespectrophotometer.Inhibitoryconstantsareobtainedthroughadditionof3.75μLTozasertibin100%DMSOorDMSOalone.Kivaluesarecalculatedasfollows,Ki=IC50/(1+[S]/Kd),where[S]=[ATP]=2.2mM,andKd(ofATPtoAbl)=70μM.ThesevaluesarecalculatedassumingaKd(ATP)of70μMforwildtypeandH396PAblkinasedomain.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[2]TheCAL-62cellsareculturedintheabsence(dimethylsulfoxide,DMSO)orthepresenceof500nMTozasertibfordifferentperiodsoftime(1-5days).Thedose-dependenteffectsofTozasertiboncell2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEproliferationareevaluatedbytreatingthedifferentATCcellsfor4dayswithdifferentconcentrationsoftheAurorainhibitor(5-500nM).Thecellsarepulselabeledwith30mMBrdUfor2hoursbeforetheendoftheincubationtime.TheBrdUincorporationisanalyzedbymeansofacolorimetricimmunoassayusingthecellproliferationELISAkit.TheresultsfromTozasertib-treatedcellsarecomparedwiththoseobservedincontrolcellsandexpressedasafoldofvariationversuscontrol.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalFortheHL-60study,femaleathymicNCr-numiceareinoculatedsubcutaneouslywith107HL-60(TB)Administration[1]leukemiacellsintotherightaxillaryarea.Treatmentisadministeredi.p.b.i.d.aftertumorsreached150−200mm3.Tozasertibispreparedinavehicleof50%PEG300in50mMphosphatebuffer.Cisplatin,formulatedinsaline,isadministeredi.p.q.4.d.foratotalofthreeinjections,atadoseof5.4mg/kg.FortheMIAPaCa-2studies,femaleMF1nudemiceareinoculatedwith107MIAPaCa-2cellsintothedorsalflank.Treatmentisadministeredi.p.b.i.d.aftertumorsreach175mm3.Tozasertibispreparedinavehicleof50%PEG300in50mMphosphatebuffer.5-fluorouracil,formulatedinsaline,isadministeredi.v.q.4.d.atadoseof50mg/kg.FortheHCT116study,femaleHsdRHrnu/nuratsareinoculatedwith107HCT116cellsintotherightflank.Treatmentisadministeredoncethetumorsreached700−950mm3.Tozasertibisadministeredcontinuouslythroughanindwellingfemoralcatheter,followedbyasalineinfusionfor4dbeforerepeatingthedosecycle.Forallstudies,tumorvolumeisdeterminedbycalipermeasurementsthreetimesaweek.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•CellDiscov.2022Sep14;8(1):92.•SciTranslMed.2018Jul18;10(450).pii:eaaq1093.•CellRep.2022Nov22;41(8):111707.•BrJPharmacol.2020Jun;177(12):2848-2859.•AmJCancerRes.2021Jun15;11(6):3021-3038.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].HarringtonEA,etal.VX-680,apotentandselectivesmallmoleculeinhibitorof