Ro-5126766-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•Agonists•ScreeningLibrarieswww.MedChemERo5126766Cat.No.:HY-18652CASNo.:946128-88-7分⼦式:C₂₁H₁₈FN₅O₅S分⼦量:471.46作⽤靶点:MEK;Raf作⽤通路:MAPK/ERKPathway储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:100mg/mL(212.11mM;Needultrasonic)扫描⼆维码，H2O:<0.1mg/mL(insoluble)运⽤溶解⽅案计算器获得适合您实验体系的溶解⽅案MassSolvent1mg5mg10mgConcentration制备储备液1mM2.1211mL10.6054mL21.2107mL5mM0.4242mL2.1211mL4.2421mL10mM0.2121mL1.0605mL2.1211mL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，并请注意储备液的保存⽅式和期限。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%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.08mg/mL(4.41mM);Clearsolution此⽅案可获得≥2.08mg/mL(4.41mM，饱和度未知)的澄溶液。以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到400μLPEG300中，混合均匀；向上述体系中加⼊50μLTween-80，混合均匀；然后继续加⼊450μL⽣理盐⽔定容⾄1mL。1/4www.MedChemEwww.MedChemE2.请依序添加每种溶剂：10%DMSO90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(4.41mM);Clearsolution此⽅案可获得≥2.08mg/mL(4.41mM，饱和度未知)的澄溶液。以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到900μL20%的SBE-β-CD⽣理盐⽔⽔溶3.液中，混合均匀。请依序添加每种溶剂：10%DMSO90%cornoilSolubility:≥2.08mg/mL(4.41mM);Clearsolution此⽅案可获得≥2.08mg/mL(4.41mM，饱和度未知)的澄溶液，此⽅案不适⽤于实验周期在半个⽉以上的实验。以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到900μL⽟⽶油中，混合均匀。BIOLOGICALACTIVITY⽣物活性Ro5126766(CH5126766)⼀种有效的双重MEK/RAF抑制剂，抑制BRAFV600E，CRAF,MEK，和BRAF，IC50分别为8.2nM,56nM,160nM和190nM。IC50&TargetMEKBRafV600EBrafCRAF160nM(IC50)8.2nM(IC50)190nM(IC50)56nM(IC50)体外研究Ro5126766(RO5126766)isanallostericinhibitorthatbindsdirectlytoMEKandpreventsitsphosphorylationbyRAFthroughtheformationofastableRAF-MEKcomplex.Ro5126766inhibitsboththephosphorylationofMEKbyRAFandtheactivationofERKbyMEK.Incell-freeMEKandRAFkinaseassays,Ro5126766effectivelyinhibitsactivationofERK2byMEK1withanIC50of160nM(SD=±0.043)andinhibitsthephosphorylationofMEK1proteinbyBRAF(IC50=190nM,SD=±0.003),BRAFV600E(IC50=8.2nM,SD=±0.0015),andCRAF(IC50=56nM,SD=±0.016).Ro5126766effectivelyinhibitsbothMEKandERKphosphorylationinapanelofhumantumorcelllinesincludingKRAS/HRASandBRAFmutantcelllinesandKRAS/HRASandBRAFwild-typecells[1].InordertoinvestigatewhetherthemevalonatepathwayaffectsthesensitivitytoMEKinhibitors,humanbreastcancerMDA-MB-231cellsharboringKRASandBRAFmutationsaretreatedRo5126766(CH5126766),withorwithoutstatins,whichinhibitsHMG-CoAreductase,therate-limitingenzymeinthemevalonatepathway.ThecombinedtreatmentofRo5126766withXU62-320demonstratesmoresignificantreductionincellgrowthinadose-dependentmannerthanthesingletreatmentofRo5126766.ThemarkedcombinedeffectsofRo5126766at40nMandXU62-320at0.3μMisalsoconfirmedonthesuppressionofthecolonyformationofthecells[2].体内研究InKRAS-mutantxenograftmodels,Ro5126766(RO5126766)inhibitsgrowthandcausestumorregressionsmoreeffectivelythananotherallostericMEKinhibitor,PD0325901.PreclinicaldatafromaseriesofhumantumormousexenograftmodelsindicatesanED50forRo5126766of0.03to0.23mg/kgandanED90of0.15to1.56mg/kg.Theseeffectivedosesareassociatedwithtargettroughconcentrationsof17to133ng/Land87to901ng/mL,respectively.[1].Inthisexperiment,Ro5126766(CH5126766)orPD0325901isadministratedattheirmaximumtolerateddose(MTD)intheHCT116model(1.5and25mg/kg,respectively).ThesedosesinhibitpERKandERKsignalingoutputatsimilardegreesinthetumorsfromthedrug-treatedmiceat4hoursfromthefirstdrugadministration.Moreover,inHCT116models,theED50forRo5126766andPD0325901are0.056and0.80mg/kg,respectively.Therefore,thedosesusedforthisexperimentare2/4www.MedChemEwww.MedChemE26.8-and31.3-foldhigherdosesthanthe50%effectivedoses,respectively.Dailyoraladministrationofeitherdrugcausessignificanttumorregressionofeachthesetumors.However,whereasinhibitionoftumorgrowthismaintainedfortheentire28-daytreatmentperiodinRo5126766-treatedmice,tumormodelsreceivingPD0325901becomerefractoryafter10daysoftreatment[3].PROTOCOLCellAssay[2]ThenumberofviablecellsisassessedwithaCellCountingKit-8assay.HumanbreastcancerMDA-MB-231cells,humanmelanomaSK-MEL-28cells,andhumannon-smallcelllungcancerA549cellsareseededatadensityof2,000cellsperwellin96-wellplatesandincubatedfor24h,andthentreatedwithRo5126766(10,20,40,and80nM)for72h.Afterafurther4hincubationwiththekitreagent,theabsorbanceat450nmofthesamplesismeasuredusingamulti-platereader[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[3]Administration[3]FemaleBALB-nu/numice(CAnN.Cg-Foxn1nu/CrlCrljnu/nu)aregivenaccesstostandardmousechowandwateradlibitum.Atotalof5×106(HCT116)or1×107(Calu-6andCOLO205)tumorcellspermouseareinjectedsubcutaneouslyintotherightflankofthe7-to9-week-oldmice.Whentumorvolumereachesto200mm3(day0),themicearerandomizedandvehicle[5%DMSOand10%2-hydroxypropyl-β-cyclodextrin(HPCD)solutionindistilledwater],Ro5126766(1.5mg/kgor2.0mg/kg)orPD0325901(25mg/kg)isadministeredorallyonceaday.Drugsareadministratedatthemaximumtolerateddose(MTD).Tumorgrowthinhibition(TGI)iscalculated.Thevalueofthe50%effectivedose(ED50)foreachcompoundiscalculated[3].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•ClinSci(Lond).2019Apr16;133(8):919-932.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].Martinez-GarciaM,etal.First-in-human,phaseIdose-escalationstudyofthesafety,pharmacokinetics,andpharmacodynamicsofRO5126766,afirst-in-classdualMEK/RAFinhibitorinpatientswithsolidtumors.ClinCancerRes.2012Sep1;18(17):4806-19.[2].Iizuka-OhashiM,etal.Bloc