Tozasertib-VX-680-DataSheet-生命科學試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemETozasertibCat.No.:HY-10161CASNo.:639089-54-6Synonyms:VX680;MK-0457分?式:C??H??N?OS分?量:464.59作?靶點:AuroraKinase;Autophagy作?通路:CellCycle/DNADamage;Epigenetics;Autophagy儲存?式:4°C,protectfromlight*Insolvent:-80°C,6months;-20°C,1month(protectfrom

light)溶解性數(shù)據(jù)體外實驗DMSO:≥106.67mg/mL(229.60mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.1524mL10.7622mL21.5244mL5mM0.4305mL2.1524mL4.3049mL10mM0.2152mL1.0762mL2.1524mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；?旦配成溶液，請分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month(protectfromlight)。-80°C儲存時，請在6個?內(nèi)使?，-20°C儲存時，請在1個?內(nèi)使?。體內(nèi)實驗請根據(jù)您的實驗動物和給藥?式選擇適當?shù)娜芙?案。以下溶解?案都請先按照InVitro?式配制澄的儲備液，再依次添加助溶劑：(為保證實驗結(jié)果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當保存；體內(nèi)實驗的?作液，建議您現(xiàn)?現(xiàn)配，當天使?；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:≥2.08mg/mL(4.48mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(4.48mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(4.48mM);ClearsolutionBIOLOGICALACTIVITY?物活性Tozasertib(VX680;MK-0457)AuroraA/B/C激酶抑制劑，Ki值分別為0.6，18，4.6nM。IC50&TargetAuroraAAuroraBAuroraC0.6nM(Ki)18nM(Ki)4.6nM(Ki)體外研究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?邊的抑制劑?師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.戶使?本產(chǎn)品發(fā)表的科研?獻?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