Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEDI-1548Cat.No.:HY-156240CASNo.:2247060-97-3分?式:C??H??N?O?S分?量:597.81作?靶點:E1/E2/E3Enzyme;Keap1-Nrf2作?通路:MetabolicEnzyme/Protease;NF-κB儲存?式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY?物活性DI-1548?種強效、選擇性、不可逆的DCN1共價抑制劑(IC50=4.6nM)。DI-1548能在納爾濃度下強效且選擇性地抑制Cullin3的neddylation修飾，對其它Cullin蛋?(包括Cullin1、2、4A、4B和5)的neddylation過程?明顯響，且?細胞毒性。DI-1548與DCN1的共價結(jié)合會破壞DCN1-UBC12相互作?，導(dǎo)致neddylation復(fù)合物崩解，進?使CRL3失活，最終導(dǎo)致NRF2積累及其靶因的上調(diào)，從?在??模型中起到保護肝臟的作?。DI-1548可?于肝損傷相關(guān)研究[1]。體外研究DI-1548(1-1000nM,24h)inhibitscullin3neddylationatconcentrationsaslowas1nMandareapproximately1000timesmorepotentthanDI-591(HY-124602)inU2OScells[1].DI-1548(0.3-1000nM,24h)inhibitstheneddylationofcullin3withover1000-foldselectivityoverothercullinmembersinvariouscelllines,includingU2OS,MDA-MB-231,KYSE70,andHCT116[1].DI-1548(0-1000nM,72h)showsnocytotoxicityinU2OS,MDA-MB-231,HCT116andKYSE70cellsatconcentrationsupto1000nM[1].DI-1548(30nM,0-48h)hasfastkineticsinselectiveinhibitionoftheneddylationofcullin3overcullin1[1].DI-1548(0.3-1000nM,1h)selectivelyandpotentlystabilizescellularDCN1inadose-dependentmanneratnanomolarconcentrations,withnoobservableeffectonDCN3[1].WesternBlotAnalysis[1]CellLine:U2OScellsConcentration:1,10,100and1000nMIncubationTime:24h1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEResult:ReducedN-Cul3levelsat1nMandwasapproximately1000timesmorepotentthanDI-591.WesternBlotAnalysis[1]CellLine:U2OS,MDA-MB-231,HCT116andKYSE70cellsConcentration:0.3,1,3,10,30,100,and1000nMIncubationTime:24hResult:SignificantlyreducedthelevelofN-CUL3atconcentrationsaslowas0.3nM,andachievedprofoundinhibitionoftheneddylationofcullin3at1-3nM.Demonstrateover300-foldgreaterpotencythanDI-591acrossvariouscelllines.Hadnoobviouseffectontheneddylationofothercullinmembersthatwereexamined,includingcullin1,2,4A,4B,and5atconcentrationsupto1000nM.InducedsignificantaccumulationofNRF2proteinatconcentrationsaslowas0.3-1nM,andwas100-1000timesmorepotentthanDI-591.HadnoobviouseffectonproteinlevelsofKEAP1andp62inU2OScells.Hadnoeffectonthelevelsofp21andBIMproteins,substratesofCRL1,andCDT1,asubstrateofCRL4A,atconcentrationsupto1000nM,consistentwithitsinabilitytoinhibitneddylationofthesecullinmembers.CellViabilityAssay[1]CellLine:U2OS,MDA-MB-231,HCT116andKYSE70cellsConcentration:0-1000nMIncubationTime:72hResult:Showednocytotoxicityinfourcancercelllinesatconcentrationsupto1000nM.WesternBlotAnalysis[1]CellLine:U2OScellsConcentration:30nMIncubationTime:5and30min,6,24,and48hResult:Inhibitedtheneddylationofcullin3within5minbuthadnoeffectontheneddylationofcullin1withtreatmenttimeupto48h.WesternBlotAnalysis[1]CellLine:U2OScells2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEConcentration:0.3,1,3,10,30,100,and1000nMIncubationTime:1hResult:EnhancedthethermalstabilityofDCN1proteininadose-dependentmanneratconcentrationsaslowas0.3nM.Exhibitedover1000-foldgreaterpotencythanDI-591inenhancingthethermalstabilityofcellularDCN1protein.ShowednodiscernibleeffectonthethermalstabilityofcellularDCN3proteinatconcentrationsupto1000nM.體內(nèi)研究DI-1548(25mg/kg,i.p.,once)elevateshepaticNRF2proteinlevelsinC57BL/6WTmice[1].AnimalModel:MaleC57BL/6WTmice(~8weeks)[1]Dosage:25mg/kgAdministration:i.p.,onceResult:InducedrobustupregulationofNRF2proteininthemouseliver.ProducedasustainedPDeffectintheliverdespitebeingclearedfromsystemiccirculationwithin3hours.REFERENCES[1].ZhouH,etal.Selectiveinhibitionofcullin3neddylationthroughcovalenttargetingDCN1protectsmicefromacetaminophen-inducedlivertoxicity.NatCommun.2021May11;12(1):2621.Mc