BFH772-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEBFH772Cat.No.:HY-100419CASNo.:890128-81-1分⼦式:C₂₃H₁₆F₃N₃O₃分⼦量:439.39作⽤靶点:VEGFR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:100mg/mL(227.59mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.2759mL11.3794mL22.7588mL5mM0.4552mL2.2759mL4.5518mL10mM0.2276mL1.1379mL2.2759mL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>>90%cornoilSolubility:≥2.5mg/mL(5.69mM);Clearsolution1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEBIOLOGICALACTIVITY⽣物活性BFH772⼀种有效的，具有⼝服活性的VEGFR2抑制剂，IC50为3nM[1]。IC50&TargetVEGFR23nM(IC50)体外研究BFH772ishighlyselective;apartfrominhibitingVEGFR2at3nMIC50,italsotargetsB-RAF,RET,andTIE-2,albeitwithatleast40-foldlowerpotency.BFH772isinactive(IC50>10μM;>2μMforcKIT)againstallothertyrosinespecific-andserine/threonine-specificproteinkinasestested.BFH772inhibitsVEGFR2withIC50of4.6±0.6nMinCHOcells.BFH772inhibitsVEGFR2withIC50of3nMinHUVECcells.BFH772inhibitstheligandinducedautophosphorylationofRET,PDGFR,andKITkinases,withIC50valuesrangingbetween30and160nM.BFH772isselective(IC50values>0.5μM)againstthekinasesofEGFR,ERBB2,INS-R,andIGF-1RandagainstthecytoplasmicBCR-ABLkinase.IC50ofBFH772([1].体内研究BFH772at3mg/kgorallydosedonceperdaypotentlyinhibitsmelanomagrowth(by54-90%forprimarytumorand71-96%formetastasisgrowth)asdepictedbytreatmenttocontrolratios.Dose–responsecurvesofBFH772at0.3,1,and3mg/kgdemonstratethatevenatthelowestconcentrations,thisnaphthalene-1-carboxamideinhibitsVEGFinducedtissueweightandTIE-2levelsbutonlyreachesstatisticalsignificanceat1mg/kgandabove[1].PROTOCOLCellAssay[1]DifferentBa/F3celllinesrenderedIL-3independentbytransductionwithvariousconstitutivelyactivetyrosinekinasesaregrowninRPMI1640mediumcontaining10%fetalcalfserum.FormaintenanceofparentalBa/F3cells,themediumisadditionallysupplementedwith10ng/mLinterleukin-3(IL-3).Forproliferationassays,Ba/F3cellsareseededon96-wellplatesintriplicatesat10000cellsperwellandincubatedwithvariousconcentrationsofcompoundsfor72hfollowedbyquantificationofviablecellsusingaresazurinsodiumsaltdyereductionreadout(commerciallyknownasAlamarBlueassay).IC50saredeterminedwiththeXLFitExcelAdd-Inusingafour-parameterdoseresponsemodel[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1]FemaleFVBmiceweighingbetween18and20garehousedingroupsofsix.PorouschamberscontainingVEGF(2μg/mL)in0.5mLof0.8%w/vagar(containingheparin,20U/mL)areimplantedsubcutaneouslyintheflankofthemice(n=6pergroup).VEGFinducesthegrowthofvascularizedtissuearoundthechamber.Thisresponseisdose-dependentandcanbequantifiedbymeasuringtheweightandTIE-2levelsofthetissue.Micearetreatedeitherorallyoncedailywithcompoundsorvehicle(PEG200100%,5mL/kg)starting4-6hbeforeimplantationofthechambersandcontinuingfor4days.Theanimalsaresacrificedformeasurementofthevascularizedtissues24hafterthelastdose.TissueweightistakenandthenalysatepreparedforTIE-2ELISAanalysis.Rats[1]2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemECathetersareimplantedintothefemoralarteryandveinofnaïvefemaleratsstrainOFAforBFH772,andBAW2881,orinthejugularveinandfemoralarteryinfemaleSprague-Dawleyratsforcompounds4,9,and10.Animalsareallowedtorecoverfor96handarehousedinsinglecageswithfreeaccesstofoodandwaterthroughouttheexperiment.FemaleOFAratsreceived2.5mg/kgofBAW2881dissolvedinethanol/dimethylisosorbide/polyethyleneglycol400/D5W(10/15/35/40v/v)or1mg/kgofBFH772dissolvedinN-methylpyrrolidone/polyethyleneglycol200(30:70,v/v)viainjectionintothefemoralvein.D5Wisglucose5%/water(v/v).Oraladministration:BAW2881andBFH772areformulatedasamicronizedsuspension(dissolved/suspendedin0.5%carboxymethylcelluloseindistilledwater)andadministeredbygavagetofemaleOFAratstodeliveradoseof25mg/kgforBAW2881or3mg/kgBFH772(n=4ratspergroup).Forcompounds4,9,and10,femaleSprague-Dawleyratsat8weeksofagereceivedanintravenousdoseof3mg/kg4,9,and10,formulatedinethanol/NMP/polyethyleneglycol400/D5W(10/10/50/30)(n=2ratspergroup),orasuspensionin0.5%carboxymethylcelluloseindistilledwaterdosedat50mg/kg(n=3ratspergroup).Attheallottedtimes,bloodsamplesarecollectedintoheparinizedtubes,andtheamountofcompoundinplasmadeterminedbyHPLC/MS-MS.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].BoldG,etal.ANovelPotentOralSeriesofVEGFR2InhibitorsAbrogateTumorGrowth