LY2109761-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemELY2109761Cat.No.:HY-12075CASNo.:700874-71-1分⼦式:C₂₆H₂₇N₅O₂分⼦量:441.52作⽤靶点:TGF-βReceptor;Autophagy作⽤通路:TGF-beta/Smad;Autophagy储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:25mg/mL(56.62mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.2649mL11.3245mL22.6490mL5mM0.4530mL2.2649mL4.5298mL10mM0.2265mL1.1325mL2.2649mL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：0.5%CMC-Na>>0.5%Tween-80Solubility:10mg/mL(22.65mM);Suspenedsolution;Needultrasonic1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE2.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.08mg/mL(4.71mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(4.71mM);Clearsolution4.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(4.71mM);Clearsolution5.请依序添加每种溶剂：5%DMSO>>40%PEG300>>5%Tween-80>>50%salineSolubility:≥1.25mg/mL(2.83mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性LY2109761可⼝服，选择性的TGF-βreceptortypeI/II抑制剂，Ki分别为38nM和300nM。IC50&TargetKi:38nM(TβRI),300nM(TβRII)体外研究LY2109761significantlyinhibitsthegrowthofL3.6pl/GLTsoftagarcoloniesinadose-dependentmanner,andresultsinappr33%inhibitionat2μMand73%inhibitionat20μM.TargetingTβRI/IIkinaseactivitywithLY2109761(5μM)almostcompletelysuppressesboththebasalandTGF-β1-stimulatedmigrationofL3.6pl/GLTcells[1].LY2109761inducesadose-dependentreductioninphosphorylationofSmad-2.HLEendogenousphosphorylationofSmad-2isinhibitedbyLY2109761.LY2109761blocksmigrationondifferentECMproteinsandinvasionofbothHLEandHLFthrougha3-dimensionalstructure.LY2109761increasesE-cadherinmRNAexpressionafter24hoursandproteinlevelsafter48hours[2].LY2109761pretreatmentreducesclonogenicsurvivalincellculturesofU87MGandT98followingradiation,resultinginanincreaseintheradiosensitivitywithaDEF0.1of1.30and1.37,respectively[3].体内研究LY2109761(50mg/kg,p.o.)greatlyreducesthetumorvolumeandincreasesthemediansurvivaldurationofthemiceto45.0days.ThemicetreatedwithLY2109761developsignificantlyfewermetastaticlesionsand,insomeofthem,nometastaticlesion,asindicatedbytheGFPsignal,canbeidentifiedintheabdomen[1].LY2109761enhancesradiation-inducedtumorgrowthdelayinaU87MGsubcutaneousxenografttumormodelinBALB/cnudemice.LY2109761increasessurvivalinanorthotopicalCSLCglioblastomamodelandenhancedantitumoractivityofradiation[2].PROTOCOLCellAssay[1]L3.6pl/GLTcellsarewashedtwicewithcoldPBSandlysedat4°CintoRIPAbuffer(50mMTrisHCl[pH8],150mMNaCl,1%NP-40,0.5%sodiumdeoxycolate,and0.1%SDS).TumorandliverspecimensarehomogenizedintoRIPAbufferwithanelectrichomogenizerandthenmaintainedinconstantagitationfortwohoursat4°C.Thelysatesareclearedbycentrifugation.Eachlysate(20µgofprotein)isseparatedby8%SDSandprobedwithpolyclonalrabbitantibodiesagainsttotalERK1/2,TβRI,andTβRIIorwithmonoclonalmouseantibodiesagainstphosphorylatedSmad2,totalSmad2,phosphorylatedERK1/2,andphosphorylatedandtotalJNK.ImmunoreactiveproteinsarevisualizedwithLumi-LightWesternblottingsubstrateaccordingtothemanufacturer'sinstructions.phosphorylatedSmad2/Smad2banddensityratios2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEarecalculatedusingtheImageQuantsoftware.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalFiftymicearerandomLyallocatedintofivegroups(n=5micepergroup)toreceivep.o.vehiclefor50μLofAdministration[1]LY2109761or50mg/kgLY2109761twiceadayp.o.Onday0,miceareanesthetizedwith1.5%isofluorane-airmixture,asmallleftabdominalflankincisioniscreated,andthespleeniscarefullyexteriorized.L3.6pl/GLTorC5LM2/GLTcells(1.0×105cells/50μLofHBSS),culturedinthepresenceofLY2109761(5μM)orDMSOfromday−5today0,areinoculatedintothespleenwitha30-gaugeneedle.Avisiblepalingofthespleenisthecriterionforsuccessfulinoculation.After10min,thespleenisremovedusingahigh-temperaturecauterytoavoidthepossibilitythattheectopicgrowthofpancreatictumorcellsinthespleencouldbeaconfoundingsourceofhematogenouslivermetastaticcells.Theabdominalwallisclosedinonelayerwithwoundclips.Treatmentwith50mg/kgLY2109761twiceadayp.o.(days1-5ofeachweekafterinoculation)iscontinuedforonegroupofuntreatedmiceinoculatedwithuntreatedcells.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•CellRes.2020Jul;30(7):610-622.•JHepatol.2015Oct;63(4):863-73.•SciTranslMed.2020Mar25;12(536):eaay8456.•NatCommun.2017Aug18;8(1):286.•Hepatology.2022Feb2.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].MelisiD,etal.LY2109761,anoveltransforminggrowthfactorbetareceptortypeIandtypeIIdualinhibitor,asatherapeuticapproachtosuppressingpancreaticcancermetastasis.MolCancerTher,