Latrepirdine-Dimebolin-生命科學試劑-MCE

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemELatrepirdineCat.No.:HY-14862CASNo.:3613-73-8Synonyms:Dimebolin;Dimebone分?式:C??H??N?分?量:319.44作?靶點:Amyloid-β;HistamineReceptor;AdrenergicReceptor;5-HTReceptor;Autophagy作?通路:NeuronalSignaling;GPCR/GProtein;Immunology/Inflammation;Autophagy儲存?式:PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY?物活性Latrepirdine?種神經(jīng)活性化合物，對組胺受體(histaminergicreceptor)，α-腎上腺素能受體(α-adrenergicreceptor)和5-羥?胺受體(serotonergicreceptor)具有拮抗活性。Latrepirdine刺激淀粉樣前體蛋?(APP)分解代謝和β-淀粉樣蛋?(amyloid-β,Aβ)分泌。IC50&TargetAmyloid-β(Aβ),Histaminergicreceptor,α-adrenergicreceptor,Serotonergicreceptor[1]體外研究Latrepirdinehasbeenreportedtopossessseveralpropertiesthatarepotentiallyrelevanttothetreatmentofneurodegenerativediseases:(1)protectionofculturedcellsfromthecytotoxicityofamyloid-β(Aβ)peptide;(2)stabilizationofmitochondrialfunctionandcalciumhomeostasis;(3)modulationofAβreleasefromculturedcells,isolatedintactnerveterminals,andfromhippocampalneuronsinlivingmousebrain;and(4)promotionofneurogenesisinthemurinehippocampus.TreatmentofculturedmammaliancellswithLatrepirdineleadstoenhancedmTOR-andAtg5-dependentautophagy.LatrepirdinemodulatesAtg5-dependentautophagicactivityinadose-dependentmannerandviathemTOR-signalingpathway.HeLacellsstablyexpressingLC3fusedaretreatedwithEGFP(eGFP-LC3)for3or6hoursintheabsenceorpresenceof50μMLatrepirdine.TreatmentwithLatrepirdinefor3or6hoursmarkedlyenhancesthenumberofeGFP-LC3punctae,indicatingthatLatrepirdineinducesformationofautophagosomes.Next,mouseN2aneuroblastomacellsaretreatedintheabsence(vehicle)orpresenceof5nM,500nMor50μMLatrepirdinefor3or6hoursinordertodeterminetheeffectsofacutedrugtreatmentontheregulationofautophagy.Asignificantanddose-dependentincreaseisobservedinLC3-IIlevelsinN2acellsfollowing3-or6-hourtreatmentwitheither500nMor50μMLatrepirdine.Asignificantdecreaseofp-mTORandp-S6KfromN2a1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEcellstreatedwith50μMLatrepirdinefor3hoursisobserved,whereasthetotalmTORandp70S6Klevelsremainrelativelyconstant[1].體內(nèi)研究LatrepirdinetreatmentofTgCRND8transgenicmiceisassociatedwithimprovedlearningbehaviorandwithareductioninaccumulationofAβ42andα-synuclein.Male,90-day-oldTgCRND8miceortheirwild-typelittermates(nTg)receive31consecutiveoncedailyi.p.injectionsofeither3.5mg/kgLatrepirdineor0.9%saline(vehicle).Attheculminationoftreatment,micearetestedforcuedandcontextualfearconditioningusingaparadigmthathasbeenwidelyacceptedforevaluatinglearningandmemorydeficitsinAPPtransgenicmice.AsignificantincreaseincuedmemoryonlyamongLatrepirdine-versusvehicle-treatedTgCRND8mice(p=0.01)isobserved.Aweak,non-significanttrendtowardanimprovementincontextualmemoryamongLatrepirdine-versusvehicle-treatedmice(p=0.099)isalsoobserved[1].PROTOCOLCellAssay[1]N2acells,stablehumancervicalcarcinoma(HeLa)cellsexpressingEGFP-LC3,andmouseembryonicfibroblasts(MEFs)derivedfromwildtypemiceorATG5-/-micearemaintainedin“growthmedium”(highglucoseDulbecco'smodifiedEagle'smediumsupplementedwith10%FBSand100units/mLPenicillin/Streptomycin)at37°C,5%CO2.N2acellsstablytransfectedwithAPPK670N,M671Laremaintainedingrowthmediumsupplementedwith0.2mg/mLG418.Cellsarewashed1×withicecoldPBS(pH7.4)thenincubatedwitheitherLatrepirdine(5nM,500nMor50μM)orvehicle(growthmedium).Following3-,6-,or24-houroftreatment,cellsarewashed1xwithicecoldPBS,andcollectedinlysisbuffer(50mMTris-HCl,150mMNaCl,1mMPepstatin,1mMPMSF,1%TritonX-100,EDTA-freemini-completeproteaseinhibitorcocktailtablet)thencentrifuged(14,000RPM)for15minutesat4°C.Fortime-courseexperiments,cellsarewashed2×withice-coldPBS(pH7.4)andincubatedfortheindicatedtimeinserum-freeDMEMcontaining50μg/mLCHXor50μg/mLCycloheximide(CHX)+50μg/mLChloroquine(CQ).Baseline(T0)samplesarecollectedimmediatelypriortotreatment[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1]Male53-55-day-oldTgCRND8mice(N=25)arerandomlydistributedintoeitherofthetwotreatmentgroups:Latrepirdine(n=13TgCRND8)orvehicle(n=12TgCRND8).Animalsreceive21consecutiveoncedailyintraperitonealinjectionsofeither3.5mg/kgLatrepirdineor0.9%saline(vehicle).90-day-oldmaleTgCRND8mice(N=28)ortheirwild-typelittermates(N=56)arerandomlydistributedintoeitheroftwotreatmentgroups:Latrepirdine(n=13TgCRND8;n=21nTg)orvehicle(n=15TgCRND8;n=25nTg).Followingtreatment,animalsaresacrificedandtranscardiallyperfusedwithice-coldPBS(pH7.4).Male90-day-old(n=5pergenotype)or120-day-old(n=6pergenotype)TgCRND8miceortheirnon-transgeniclittermatesaresacrificedandtranscardiallyperfusedwithice-coldPBS(pH7.4).Onehemispherefromeachmouseispost-fixedin4%paraformaldeyhdeinPBS(pH7.4)forhistologicalanalysisandtheotherhemisphere