Hotline:400-820-3792Inhibitors ? ScreeningLibraries ? Proteinswww.MedChemEBY13Cat.No.:HY-176225分子式:C??H??N?O?分子量:645.71作用靶點:PROTACs;Src;EstrogenReceptor/ERR;Apoptosis作用通路:PROTAC;ProteinTyrosineKinase/RTK;VitaminDRelated/NuclearReceptor;Apoptosis儲存方式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY生物活性BY13是一種SRC-3PROTAC降解劑，其DC50為0.031μM。BY13通過下調ERα水平選擇性阻斷ER信號通路，其高于對雄激素受體(AR)的選擇性。BY13通過誘導細胞周期停滯于G1期并誘導細胞凋亡(apoptosis)，有效克服乳腺癌的內分泌耐藥性，其作用優(yōu)于Fulvestrant(HY-13636)。BY13顯著抑制LCC2異種移植小鼠模型中耐藥乳腺腫瘤的生長，且無明顯毒性[1]。粉色：SRC-3ligand(SI-2)(HY-101447)；藍色：CRBNligaseligand(HY-41547)；黑色：linker(HY-176226)IC50&TargetERαSRC-30.031μM(DC50)體外研究BY13(0.1-10μM,24h)dose-dependentlyreducesSRC-3andERαproteinlevelinMCF-7cellswith71%and85%degradation,respectively[1].BY13(0.01-10μM,36h)potentlyinhibitsproliferationofwild-type,mutant-typeanddrug-resistantbreastcancercells(IC50of0.003-0.35μMforMCF-7,LCC2andMCF-7D538G/Y537S/EGFRcells),anddose-dependentlydegradesSRC-3andERαinmutant-typecells(particularlyinMCF-7Y537Scellsat0.1μM)[1].BY13(0.1-10μM,3-48h)effectivelyreducesSRC-3andERαproteinlevel,superiortoFulvestrant(HY-13636),andreachesthemaximaldegradationafter36hwhilealmostnolongerincreasewithtimeinLCC2cells[1].BY13(0.01-20μM,36h)dramaticallydecreasestheproteinlevelofSRC-3(0.1μMonly)andERα(1μMonly)(DC50of0.031μMforSRC-3)withsubtypeselectivityfavoringSRC-1overSRC-2inMCF-7cells[1].BY13(0.01-5μM,24h)down-regulatestheproteinlevelofARwhilethiseffectisweakercomparedtoERαinMCF-7cells,andmoderatelyinhibitsAR-overexpressingLNCaPcells(IC50:1.43μM)[1].BY13(1μM,36h)significantlyreducesSRC-3proteinleveldependingonaubiquitinproteasomesystem(UPS)pathwayinMCF-7cells(similarphenomenaobservedinLCC2cells)[1].1/4 MasterofBioactiveMolecules—您身邊的抑制劑大師www.MedChemEBY13(1μM,6h,40-76℃)canentertumorcellsanddirectlybindtoSRC-3withsignificantlyenhancementoftheSRC-3proteinthermalstabilityathightemperatureinMCF-7cells[1].BY13(10μM,6h)inducesthespatialproximityofSRC-3andCRBN,followedbypromotingtheformationoftheSRC-3-BY13-CRBNternarycomplexinMCF-7cells[1].BY13(0.01-10μM,36h)significantlyincreasesthemRNAexpressionofSRC-3withtheincreasingconcentrationsandeffectivelyreducesthemRNAlevelofERαinLCC2cells[1].BY13(1-20μM,48h)significantlyinducesbreastcancercellsapoptosiswithenhancementofbothearlyandlateapoptosisinMCF-7andLCC2cells.BY13(5-20μM,48h)dose-dependentlyarrestsLCC2cellsintheG1phase,andthecellproportionissignificantlyhigherthanthatintheSphase.BY13(0.01-100μM)hassufficientmetabolicstabilityandanacceptablesafetyprofilewithIC50of2.73and1.1μMforCYP3A4andhERGchannel.WesternBlotAnalysis[1]CellLine:MCF-7cells,LCC2cellsConcentration:0.01,0.1,1,5,10μMIncubationTime:24hResult:DegradedbothSRC-3andERαproteininMCF-7cellsat1μMwith71%and85%degradation,respectively.Dose-dependentlyreducedSRC-3andERαproteinlevelinMCF-7cellswithoutacanonical“hookeffect”aswellassignificantlydegradesSRC-3andERαinLCC2cells,superiortoFulvestrant(HY-13636).Down-regulatedtheproteinlevelofARwhilethiseffectisweakercomparedtoERαinMCF-7cellsWesternBlotAnalysis[1]CellLine:MCF-7cells,LCC2cellsConcentration:1μMIncubationTime:MCF-7cells(3,6,9,12,24,36h),LCC2cells(3,6,9,12,24,36,48h)Result:EffectivelyreducedtheSRC-3proteinlevelafter24handreachedthemaximaldegradationinMCF-7cellsat36h.SignificantlyreducedtheproteinlevelofSRC-3afteronly9hoftreatment,andreachedthemaximaldegradationinLCC2cellsat36hwithalmostnolongerincreaseofdegradation.WesternBlotAnalysis[1]CellLine:MCF-7cells,LCC2cellsConcentration:1μMafter1μMofMG-132,Bortezomib,Chloroquineor(BY13-Neg,SI-2and2/4 MasterofBioactiveMolecules—您身邊的抑制劑大師www.MedChemEPomalidomide)for2hIncubationTime:36hResult:SignificantlyreducedSRC-3proteinleveldependingonaubiquitinproteasomesystem(UPS)pathway,butthiseffectwasreversedbytwoproteasomeinhibitors(MG-132,BortezomibandSI-2)notbyBY13-NeginMCF-7cells(similarsymptomsobservedinLCC2cells).ApoptosisAnalysis[1]CellLine:MCF-7cells,LCC2cellsConcentration:1,5,10,20μMIncubationTime:48hResult:EffectivelyinducedcellapoptosiswithenhancementofbothearlyandlateapoptosisinMCF-7cellsatincreasedconcentrations(35%maximalearlyapoptosisrate).Mainlyinducedthelateapoptosisat1μM,butresultedinearlyapoptosisat10μMandinducedastrongearlyapoptoticeffectat20μM(＞30%earlyapoptosisrate)inLCC2cells.體內研究BY13(3-10μM/kg,i.p.,onceeverytwodaysfor23?days)hasstrongantiendocrine-resistantactivitythroughtargetingtheSRC-3andERαdegradationwithahighsafetyproperty,andimprovesthepoorprognosisofendocrine-resistantbreastcancerinLCC2xenograftmicemodel[1].AnimalModel:FemaleBalb/cnudemice(4weeksold)wereinjectedsubcutaneouslyintotherightloweraxillawithLCC2cells(5?×?106cells/mouse)toinduceLCC2xenograftmicemodel[1].Dosage:3,510μM/kg,Tamoxifen(10μM/kg)andFulvestrant(5μM/kg)Administration:i.p.,onceeverytwodaysfor23?daysaftertumorsreachedapproximately100?mm3,andthencollectedsampleat36day1.Result:Significantlyinhibitedtumorgrowthat3μM/kg,withsuperioreffectof54%tumorgrowthinhibitionrate54%at5μM/kg.AlmostcompletelydegradedSRC-3proteinwithinthetumortissuesat3μM/kg,andtheproteinlevelsofbothSRC-3andERαwerefurtherdecreasedat5μM/kg.Hadawidetherapeuticwindow,withouteffectweightgainofmicemodelupto10μM/kg.Hadahighsafetypropertywithoutobservablehistopathologicalchangesintheorgantissuesofmicemodelduringtheadministrationperi