BFH772-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEBFH772Cat.No.:HY-100419CASNo.:890128-81-1分?式:C??H??F?N?O?分?量:439.39作?靶點(diǎn):VEGFR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(yàn)DMSO:100mg/mL(227.59mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.2759mL11.3794mL22.7588mL5mM0.4552mL2.2759mL4.5518mL10mM0.2276mL1.1379mL2.2759mL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(shí)，請?jiān)?個(gè)?內(nèi)使?，-20°C儲存時(shí)，請?jiān)?個(gè)?內(nèi)使?。體內(nèi)實(shí)驗(yàn)請根據(jù)您的實(shí)驗(yàn)動(dòng)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(yàn)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(yàn)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%DMSO>>90%cornoilSolubility:≥2.5mg/mL(5.69mM);Clearsolution1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEBIOLOGICALACTIVITY?物活性BFH772?種有效的，具有?服活性的VEGFR2抑制劑，IC50為3nM[1]。IC50&TargetVEGFR23nM(IC50)體外研究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].體內(nèi)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?邊的抑制劑?師www.MedChemECathetersareimplantedintothefemoralarteryandveinofna?vefemaleratsstrainOFAforBFH772,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