VX-11e-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEVX-11eCat.No.:HY-14178CASNo.:896720-20-0分⼦式:C₂₄H₂₀Cl₂FN₅O₂分⼦量:500.35作⽤靶点:ERK作⽤通路:MAPK/ERKPathway;StemCell/Wnt储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:≥100mg/mL(199.86mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制备储备液1mM1.9986mL9.9930mL19.9860mL5mM0.3997mL1.9986mL3.9972mL10mM0.1999mL0.9993mL1.9986mL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。-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:≥3.25mg/mL(6.50mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥3.25mg/mL(6.50mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性VX-11e⼀种⾼效的，具有选择性的，可⼝服的ERK抑制剂，Ki值<2nM。IC50&TargetERK2GSK3CDK2AURA2nM(Ki)395(Ki)852(Ki)540(Ki)体外研究VX-11eisactiveintheHT29cellproliferationassay(IC50=48nM)[1].体内研究VX-11eisorallybioavailableinbothratandmice[1].VX-11e(50mg/kg,p.o.)resultsinrobustinhibitionofpRSK,andinhibitstumorgrowthinNSGmicebearinghumanmelanomaRPDXtumors.VX-11ewithBKM120significantlyimprovestumorgrowthinhibition[2].PROTOCOLKinaseAssay[1]CompoundsareassayedfortheinhibitionofERK2byaspectophotometriccoupled-enzymeassay.Inthisassay,afixedconcentrationofactivatedERK2(10nM)isincubatedwithvariousconcentrationsofthecompoundsinDMSO(2.5%)for10min.at30°Cin0.1mol/LHEPESbuffer,pH=7.5,containing10mMMgCl2,2.5mMphosphoenolpyruvate,200μMNADH,150μg/mLpyruvatekinase,50μg/mLlactatedehydrogenaseand200μMerktidepeptide.Thereactionisinitiatedbytheadditionof65μMATP.Therateofdecreaseofabsorbanceat340nMismonitored.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]Cellproliferationismeasuredby3H-thymidineincorporation.Thecellsareplatedataconcentrationof10,000cells/wellina96-wellplateusinggrowthmedia,RPMI1640containing10%FBS.Seriallydilutedcompoundsareadded.Thecellsandcompoundsareincubatedfor48hoursat37°Cincubator.After48hours,0.4μCiof3H-thymidineisaddedtoeachwellsfor8hoursandreturnedtothe37°Cincubator.ThecellsareharvestedusingaTomtec96-wellcellharvesterandtheCPMisdeterminedusingtheWallac1205BETAPLATEliquidscintillationcounter.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalHumanmelanomaRPDXtumorsareexpandedinvivousingNSGmicepriortothetherapyexperiments.Administration[2]Pooledtumorchunksbankedfromearlymousepassagesareimplantedinto50NSGmice(1:10expansion).Thesetumorsareharvestedwhenreachingthemaximumvolumeallowedontheprotocol(1,000mm3),digested,andbankedaslivecells.Thelargerpartofthisstockisretainedasamasterbank,andtheotherpartisimplantedata1:5ratiointoNSGmicetouseinthetherapyexperiments.TheexpansionphaseisundercontinuousdrugpressurewithPLX4720200ppmchemicaladditivedietatapproximatelyclinicalplasmalevels.TheplasmalevelsofPLX4720(103.7μg/mL±3.2after7days)aresimilartosteady-state2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemElevelsinpatientstreatedwithvemurafenib960mgtwiceaday(130.6μg/mL±71.78).Whentumorshavereached200mm3percalipermeasurement,animalsarerandomizedintotreatmentgroupsfollowedbya3-dayishoutphase.Tumorsizeisassessedtwiceweeklypercalipermeasurement.Micearesacrificedaftertwoweeksoftreatmentorwhennecessaryforanimalwelfare.Dosingisprolongedwhentumorcontrolisachievedasindicated.Tumortissueisconservedinformalin(forFFPE)andsnap-frozeninliquidN2forproteinextraction.Treatmentgroupsaresacrificed4hoursafterlastdose.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•SciTranslMed.2018Jul18;10(450).pii:eaaq1093.•CellDeathDis.2022May12;13(5):451.•JInvestDermatol.2020Sep9;S0022-202X(20)32055-8.•StemCellsDev.2020Jul1;29(13):863-875.•ACSCombSci.2019Dec9;21(12):805-816.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].Aronov,AlexM.,etal.Structure-GuidedDesignofPotentandSelectivePyrimidylpyrroleInhibitorsofExtracellularSignal-RegulatedKinase(ERK)UsingConformationalControl.JournalofMedicinalChemistry(2009),52(20),6362-6368.[2].KreplerC,etal.PersonalizedPreclinicalTrialsinBRAFInhibitor-ResistantPatient-DerivedXenograftModelsIdentifySecond-LineCombinationTherapie