Foretinib-XL880-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEForetinibCat.No.:HY-10338CASNo.:849217-64-7Synonyms:XL880;GSK1363089;GSK089;EXEL-2880分?式:C??H??F?N?O?分?量:632.65作?靶點(diǎn):VEGFR;c-Met/HGFR作?通路:ProteinTyrosineKinase/RTK儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗DMSO:≥38mg/mL(60.06mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM1.5807mL7.9033mL15.8065mL5mM0.3161mL1.5807mL3.1613mL10mM0.1581mL0.7903mL1.5807mL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。-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(shí)驗請根據(jù)您的實(shí)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(shí)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(shí)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)1/4MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE1.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(3.95mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(3.95mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(3.95mM);ClearsolutionBIOLOGICALACTIVITY?物活性Foretinib?種多靶點(diǎn)酪氨酸激酶抑制劑，抑制Met和KDR的IC50分別為0.4nM和0.9nM。IC50&TargetKDRc-Met0.9nM(IC50)0.4nM(IC50)體外研究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].體內(nèi)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?邊的抑制劑?師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.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?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?邊的抑制劑?師www.MedChemEREFERENCES[1].Qi