GPX4-AUTAC-生命科学试剂-MCE

Hotline:400-820-3792Inhibitors • ScreeningLibraries • Proteinswww.MedChemEGPX4-AUTACCat.No.:HY-176220分子式: C₅₀H₅₇Cl₂FN₁₂O₉S₂分子量: 1124.1作用靶点: AUTACs;Autophagy;GlutathionePeroxidase;Ferroptosis作用通路: Autophagy;PROTAC;Apoptosis;MetabolicEnzyme/Protease储存方式: PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY生物活性GPX4-AUTAC是一种靶向GPX4的自噬介导降解剂(AUTAC)。GPX4-AUTAC由抑制剂ML162-yne(HY-153748)、降解标签FBnG(HY-W073762)和glycollinker(HY-W021401)组成。GPX4-AUTAC能够促进E3连接酶TRAF6对GPX4的泛素化修饰，并增强其与GPX4和p62的结合，从而实现GPX4的选择性自噬依赖性降解。GPX4-AUTAC显著诱导铁死亡(ferroptosis)，并在乳腺癌细胞、乳腺癌来源的类器官(PDOs)和MDA-MB-231肿瘤异种移植小鼠模型中表现出强大的抗癌活性，与Sulfasalazine(SAS)(HY-14655)或化疗药物(Paclitaxel(HY-B0015)或Cisplatin(HY-17394))联合使用时具有强效协同作用[1]。体外研究GPX4-AUTAC(5-80μM,12-72h)significantlyreducestheproteinlevelofGPX4inadose-andtime-dependentmanner,butbarelyaffectsthemRNAlevelofGPX4inMDA-MB-231andMCF-7cells[1].GPX4-AUTAC(20-40μM,24h)effectivelyreducestheproteinlevelofGPX4inovariancancer,lungcancer,melanoma,andgliomacells[1].GPX4-AUTAC(10μM,24-96h,37-57℃)continuouslydown-regulatesGPX4proteinlevelfor96handsignificantlyenhancesthethermalstabilityofGPX4inheat-denaturedintactcellsandcelllysatesinMDA-MB-231andMCF-7cells[1].GPX4-AUTAC(5-80μM,24h)selectivelydegradesGPX4withoutinfluenceotherselenoproteinsexpression,suchasofGPX1,TXNRD1(GPX4homologs)orHK2(aTRAF6/p62substrate)[1].GPX4-AUTACselectivelydegradesGPX4andinducesferroptosisintumorcells(MDA-MB-231cells)withminimaleffectonnormalcells(MCF-10Acells)[1].GPX4-AUTAC(10μM,24h)autophagy-dependentlyacceleratesGPX4degradation,increasespolyubiquitylationofGPX4,specificallytheendogenousandexogenousK63-linkedubiquitinchainsinHEK293Tcells[1].GPX4-AUTAC(10-40μM,12h)isreversedbyPYR-41,NH4Cland3-MAtoreducethedown-regulationofGPX4inMDA-MB-231cells[1].1/5 MasterofBioactiveMolecules—您身边的抑制剂大师www.MedChemEGPX4-AUTAC(24h)inducesco-localizationofGPX4andp62andincreasesco-localizationofGPX4withLC3BorLAMP2inPDOs[1].GPX4-AUTAC(40 μM,24 h)selectivelytargetsGPX4andferroptosisissignificantlyenrichedinMDA-MB-231cells[1].GPX4-AUTAC(5-40μM,72h)hasaspecificferroptosisinductiveeffect,andsignificantlyenhancedthelipidROSandpromotedaccumulationofFe2+inMDA-MB-231cells,butonlyfullyrescuedbytheferroptosisinhibitorFerrostatin-1(Fer-1)(HY-100579)inMCF-7cells[1].GPX4-AUTAC(5-80μM)inducesferroptosiswithmitochondrialdysfunction(abnormalmorphology,densecontentanddisruptedcrista)andsignificantincreaseinmRNAlevelsofPTGS2inMDA-MB-231[1].GPX4-AUTAC(5-40μM,24-96h)dose-andtime-dependentlyinhibitedcellviabilityandproliferation,withinsensitivitytoknock-downofGPX4inbothMDA-MB-231andMCF-7cells[1].GPX4-AUTAC(5-40μM,4days)significantlyinhibitsthegrowthofPDOswithreducingdiameterandbright-field,andPDOswithhighexpressionofGPX4increasessensitivity[1].GPX4-AUTAC(10-20μM,4days)down-regulatesGPX4expressionandinducesferroptosiswithhighlevelof4-HNE,andinhibitscancercellproliferationinPDOswithlowlevelofKi67[1].GPX4-AUTACinducesmuchmoreferroptosisandhasastrongeranti-cancereffectincombinationwithSulfasalazinecomparedtoSulfasalazinealonetreatmentinMDA-MB-231andMCF-7cellsandPDOs[1].GPX4-AUTAC(10 μM,72hor4days)sensitizesMDA-MB-231andHCC1806cellstochemotherapyandsynergisticallyinhibitscellviabilityandproliferationandthegrowthofPDOsincombinationwithchemotherapydrugs(PaclitaxelorCisplatin)[1].CellViabilityAssay[1]CellLine:MCF-7cellsConcentration:5,10,20,40μMIncubationTime:72hafterFer-1,Z-VAD-FMK,Nec-1,OlapariborVX7651hResult:Hadaspecificferroptosisinductiveeffect,onlyfullyrescuedbytheferroptosisinhibitorferrostatin-1(Fer-1),butnotbytheinhibitorofapoptosis(Z-VAD-FMK),necroptosis(Nec-1),parthanatos(Olaparib),orpyroptosis(VX765)inMCF-7cells.CellViabilityAssay[1]CellLine:MDA-MB-231cells,MCF-7cellsConcentration:MDA-MB-231cells,MCF-7cells(5,10,20,40μM),shNT,shGPX4cells(10,20,40,80μM)IncubationTime:72hResult:Dose-dependentlyinhibitedtumorcellviabilityinbothMDA-MB-231,MCF-7andshGPX4cells.CellProliferationAssay[1]2/5 MasterofBioactiveMolecules—您身边的抑制剂大师www.MedChemECellLine:MDA-MB-231cells,MCF-7cellsConcentration:5,10μMIncubationTime:24,48,72,96hResult:Dose-andtime-dependentlyinhibitedtumorcellproliferationinbothMDA-MB-231andMCF-7cells.WesternBlotAnalysis[1]CellLine:MDA-MB-231cells,MCF-7cells,MCF-10AcellsConcentration:MDA-MB-231cells,MCF-7cells,MCF-10Acells(5,10,20,40,80μM)IncubationTime:12,24,48,72hResult:SignificantlyreducedtheexpressionofGPX4proteininadose-andtime-dependentmanner.SignificantlyreducedtheexpressionofGPX4inMDA-MB-231cellswithminimaleffectonMCF-10Acellsat24h.WesternBlotAnalysis[1]CellLine:MDA-MB-231cells,MCF-7cellsConcentration:10μMIncubationTime:24,48,72,96hResult:Continuouslydown-regulatedGPX4proteinlevelfor96hafterwash-outinMDA-MB-231andMCF-7cells.SignificantlyenhancedthethermalstabilityofGPX4inheat-denaturedintactcellsandcelllysatesinMDA-MB-231andMCF-7cellsat37-57℃and24h.WesternBlotAnalysis[1]CellLine:MDA-MB-231cells,HEK293TcellsConcentration:10,40μMIncubationTime:12hResult:SignificantlyacceleratedtheturnoverrateofGPX4inMDA-MB-231cellswhenaddedCHXat10μM.Down-regulatedGPX4expression,butthiseffectreversedbyPYR-41(aninhibitorofcell-permeableubiquitinE1enzyme)inMDA-MB-231cellsat10μM.Enhancedthedown-regulationofGPX4promotedbyoverexpressionofp62,butrescued3/5 MasterofBioactiveMolecules—您身边的抑制剂大师www.MedChemEbyknockdownofp62inHEK293Tcellsat40μM.RealTimeqPCR[1]CellLine:MDA-MB-231cells,MCF-7cellsConcentration:5,10,20,40,80μMIncubationTime:24hResult:Playedaregulatoryroleatthepost-translationallevel,andbarelyaffectedthemRNAlevelofGPX4inbothMDA-MB-231andMCF-7cells.Immunofluorescence[1]CellLine:MDA-MB-231cellsConcentration:5,10,20,40μMIncubationTime:72hResult:SignificantlyenhancedthelipidROSandpromotedaccumulationofFe<>sup2+inMDA-MB-231cells.体内研究GPX4-AUTAC(10-20mg/kg,i.p.,dailyfor10-12days)hasapotentanti-canceractivity,andsignificantlyreducestumorweightsandvolumes,GPX4proteinlevels,andpercentageofKi67whileincreasing4-HNElevelswithnosignificanttoxicityinvitalorgans,throughpreferentiallyaccumulationandselectivelydegradationofGPX4intumortissuesofMDA-MB-231tumorxenograftmicemodel(minimaleffectonnormaltissues)[1].GPX4-AUTAC(10-20mg/kgcombinedSulfasalazine100 mg/kg,i.p.,dailyfor10-12days)synergizeswithSulfasalazinetoexertstrongtumorsuppressiveactivityviainducingferroptosiswithnomeasurabletoxicityinMDA-MB-231tumorxenograftmicemodel[1].AnimalModel:FemaleBALB/cnudemice(4-6weeksold)wereinjectedsubcutaneouslyintotherightflankwithMDA-MB-231cells(5 × 106cells/mouse)[1].Dosage:10,20mg/kgAdministration:i.p.,dailyfor10-12dayswhenthetumorsizereached50-100 mmResult:SelectivelydegradedGPX4intumorstissues,butnotinthenormaltissuesincludingtheheart,lung,liver,spleen,andkidney.SignificantlydecreasedtumorweightsandvolumesinMDA-MB-231tumorxenograftmicemodel.MarkedlydecreasedtheproteinlevelofGPX4inMDA-MB-231tumorxenograftmicemodel.4/5 MasterofBioactiveMolecules—您身边的抑制剂大师www.MedChemEEffectivelydecreasedpercentageofKi67,butincreasedpercentageofKi674-HNEinMDA-MB-231tumorxenograftmicemodel.AnimalModel:FemaleBALB/cnudemice(4-6weeksold)were