Ro-5126766-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?Agonists?ScreeningLibrarieswww.MedChemERo5126766Cat.No.:HY-18652CASNo.:946128-88-7分?式:C??H??FN?O?S分?量:471.46作?靶點:MEK;Raf作?通路:MAPK/ERKPathway儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:100mg/mL(212.11mM;Needultrasonic)掃描?維碼，H2O:<0.1mg/mL(insoluble)運(yùn)?溶解?案計算器獲得適合您實驗體系的溶解?案MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.1211mL10.6054mL21.2107mL5mM0.4242mL2.1211mL4.2421mL10mM0.2121mL1.0605mL2.1211mL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，并請注意儲備液的保存?式和期限。體內(nèi)實驗請根據(jù)您的實驗動物和給藥?式選擇適當(dāng)?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(dāng)保存；體內(nèi)實驗的?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%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.08mg/mL(4.41mM);Clearsolution此?案可獲得≥2.08mg/mL(4.41mM，飽和度未知)的澄溶液。以1mL?作液為例，取100μL20.8mg/mL的澄DMSO儲備液加到400μLPEG300中，混合均勻；向上述體系中加?50μLTween-80，混合均勻；然后繼續(xù)加?450μL?理鹽?定容?1mL。1/4www.MedChemEwww.MedChemE2.請依序添加每種溶劑：10%DMSO90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(4.41mM);Clearsolution此?案可獲得≥2.08mg/mL(4.41mM，飽和度未知)的澄溶液。以1mL?作液為例，取100μL20.8mg/mL的澄DMSO儲備液加到900μL20%的SBE-β-CD?理鹽??溶3.液中，混合均勻。請依序添加每種溶劑：10%DMSO90%cornoilSolubility:≥2.08mg/mL(4.41mM);Clearsolution此?案可獲得≥2.08mg/mL(4.41mM，飽和度未知)的澄溶液，此?案不適?于實驗周期在半個?以上的實驗。以1mL?作液為例，取100μL20.8mg/mL的澄DMSO儲備液加到900μL??油中，混合均勻。BIOLOGICALACTIVITY?物活性Ro5126766(CH5126766)?種有效的雙重MEK/RAF抑制劑，抑制BRAFV600E，CRAF,MEK，和BRAF，IC50分別為8.2nM,56nM,160nM和190nM。IC50&TargetMEKBRafV600EBrafCRAF160nM(IC50)8.2nM(IC50)190nM(IC50)56nM(IC50)體外研究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].體內(nèi)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.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?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