Foretinib-XL880-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEForetinibCat.No.:HY-10338CASNo.:849217-64-7Synonyms:XL880;GSK1363089;GSK089;EXEL-2880分⼦式:C₃₄H₃₄F₂N₄O₆分⼦量:632.65作⽤靶点:VEGFR;c-Met/HGFR作⽤通路:ProteinTyrosineKinase/RTK储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:≥38mg/mL(60.06mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制备储备液1mM1.5807mL7.9033mL15.8065mL5mM0.3161mL1.5807mL3.1613mL10mM0.1581mL0.7903mL1.5807mL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)1/4MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE1.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(3.95mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(3.95mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(3.95mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性Foretinib⼀种多靶点酪氨酸激酶抑制剂，抑制Met和KDR的IC50分别为0.4nM和0.9nM。IC50&TargetKDRc-Met0.9nM(IC50)0.4nM(IC50)体外研究ForetinibinhibitsHGFreceptorfamilytyrosinekinaseswithIC50valuesof0.4nMforMetand3nMforRon.ForetinibalsoinhibitsKDR,Flt-1,andFlt-4withIC50valuesof0.9nM,6.8nMand2.8nM,respectively.ForetinibinhibitscolonygrowthofB16F10,A549andHT29cellswithIC50of40nM,29nMand165nM,respectively[1].ArecentstudyindicatesForetinibaffectscellgrowthdifferentlyingastriccancercelllinesMKN-45andKATO-III.ForetinibinhibitsphosphorylationofMETanddownstreamsignalingmoleculesinMKN-45cells,whiletargetsGFGR2inKATO-IIIcells[2].体内研究Foretinib(100mg/kg,p.o.)resultsinsubstantialinhibitionofphosphorylationofB16F10tumorMetandligand(e.g.,HGForVEGF)-inducedreceptorphosphorylationofMetinliverandFlk-1/KDRinlung,whichbothpersistthrough24hours.Foretinib(30-100mg/kg,oncedaily,p.o.)resultsinreductionintumorburden.Thelungsurfacetumorburdenisreducedby50%and58%followingtreatmentwith30and100mg/kgForetinib,respectively.ForetinibtreatmentofmicebearingB16F10solidtumorsalsoresultsindose-dependenttumorgrowthinhibitionof64%and87%at30and100mg/kg,respectively.Forbothstudies,administrationofForetinibiswelltoleratedwithnosignificantbodyweightloss[1].ForetinibisdevelopedtotargetabnormalsignalingofHGFthroughMetandsimultaneouslytargetseveralreceptorstyrosinekinaseinvolvedintumorangiogenesis.Foretinibcausestumorhemorrhageandnecrosisinhumanxenograftswithin2to4hours,andmaximaltumornecrosisisobservedat96hours(afterfivedailydoses),resultingincompleteregression[3].PROTOCOLKinaseAssay[1]Kinaseinhibitionisinvestigatedusingoneofthreeassayformats:[33P]phosphoryltransfer,luciferase-coupledchemiluminescence,orAlphaScreentyrosinekinasetechnology.IC50sarecalculatedbynonlinearregressionanalysisusingXLFit.33P-PhosphorylTransferKinaseAssayReactionsareperformedin384-wellwhite,clearbottom,high-bindingmicrotiterplates.Platesarecoatedwith2μg/wellofproteinorpeptidesubstrateina50μLvolumeofcoatingbuffercontained40μg/mLsubstrate(poly(Glu,Tyr)4:1,22.5mMNa2CO3,27.5mMNaHCO3,50mMNaCland3mMNaN3.Coatedplatesarewashedoncewith50μLofassaybufferfollowingovernightincubationatroomtemperature(RT).Testcompoundsandenzymesarecombinedwith33P-γ-ATP(3.3μCi/nmol)inatotalvolumeof20μL.ThereactionmixtureisincubatedatRT2/4MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEfor2hoursandterminatedbyaspiration.Themicrotiterplatesaresubsequentlywashed6timeswith0.05%Tween-PBSbuffer(PBST).Scintillationfluid(50μL/well)isaddedandincorporated33PismeasuredbyliquidscintillationspectrometryusingaMicroBetascintillationcounter.Luciferase-CoupledChemiluminescenceAssayReactionsareconductedin384-wellwhite,mediumbindingmicrotiterplates.Inafirststepenzymeandcompoundarecombinedandincubatedfor60minutes;reactionsareinitiatedbyadditionofATPandpeptidesubstrate(poly(Glu,Tyr)4:1)inafinalvoumeof20μL,andincubatedatRTfor2-4hours.Followingthekinasereaction,a20μLaliquotofKinaseGloisaddedandluminescencesignalismeasuredusingaVictorplatereader.TotalATPconsumptionislimitedto50%.AlphaScreenTMTyrosineKinaseAssayDonorbeadscoatedwithstreptavidinandacceptorbeadscoatedwithPY100anti-phosphotyrosineantibodyareused.Biotinylatedpoly(Glu,Tyr)4:1isusedasthesubstrate.Substratephosphorylationismeasuredbyadditionofdonor/acceptorbeadsbyluminescencefollowingdonor-acceptorbeadcomplexformation.Kinaseandtestcompoundsarecombinedandpreincubatedfor60minutes,followedbyadditionofATP,andbiotinylatedpoly(Glu,Tyr)inatotalvolumeof20μLin384-wellwhite,mediumbindingmicrotiterplates.Reactionmixturesareincubatedfor1houratroomtemperature.Reactionsarequenchedbyadditionof10μLof15-30μg/mLAlphaScreenbeadsuspensioncontaining75mMHepes,pH7.4,300mMNaCl,120mMEDTA,0.3%BSAand0.03%Tween-20.After2-16hoursincubationatroomtemperatureplatesarereadusinganAlphaQuestreader.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]B16F10,A549,andHT29cells(1.2×103perwell)aremixedwithsoftagarandseededina96-wellplatecontaining10%FBSandEXEL-2880overabaseagarlayer.Fornormoxicconditions,theplatesareincubated(37°C)for12to14daysin21%oxygen,5%CO2,and74%nitrogen,whereasincubation(37°C)underhypoxicconditionsisdoneinahypoxiachamberin1%oxygen,5%CO2,and94%nitrogen.Thenumberofcoloniesisevaluatedundereachconditionfollowingadditionof50%AlamarBlueandfluorescencedetection.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalInvivotargetmodulationstudiesaredoneinnaivemiceormicebearingB16F10tumors.ForetiniborvehicleAdministration[1](0.9%normalsaline)isadministeredat10mL/kgviaoralgavage.ForexaminationofMetphosphorylationinliver,HGF(10μg/mouse)isadministeredi.v.10minbeforeharvest.ForexaminationofFlk-1/KDRphosphorylationinlung,VEGF(10μg/mouse)isadministeredi.v.30minbeforeharvest0.5hlater.Receptorphosphorylationanalysisisdeterminedbyimmunoblotanalysis.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•CancerDiscov.2021Jan;11(1):126-141.•NatBiomedEng.2018Aug;2(8):578-588.•SciTranslMed.2018Jul18;10(450).pii:eaaq1093.•CellChemBiol.2018Feb15;25(2):206-214.e11.•RSCAdv.2019,9,4862-4869Seemorecustomervalidationsonwww.MedChemE3/4MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEREFERENCES[1].Qi