BMS-299897-DataSheet-生命科學試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEBMS299897Cat.No.:HY-50883CASNo.:290315-45-6分?式:C??H??ClF?NO?S分?量:511.94作?靶點:γ-secretase作?通路:NeuronalSignaling;StemCell/Wnt儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:≥30mg/mL(58.60mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM1.9534mL9.7668mL19.5335mL5mM0.3907mL1.9534mL3.9067mL10mM0.1953mL0.9767mL1.9534mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；?旦配成溶液，請分裝保存，避免反復凍融造成的產(chǎn)品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲存時，請在6個?內(nèi)使?，-20°C儲存時，請在1個?內(nèi)使?。體內(nèi)實驗請根據(jù)您的實驗動物和給藥?式選擇適當?shù)娜芙?案。以下溶解?案都請先按照InVitro?式配制澄的儲備液，再依次添加助溶劑：(為保證實驗結(jié)果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當保存；體內(nèi)實驗的?作液，建議您現(xiàn)?現(xiàn)配，當天使?；以下溶劑前顯?的百分?指該溶劑在您配制終溶液中的體積占?；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的?式助溶)1.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%saline1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemESolubility:≥2.5mg/mL(4.88mM);ClearsolutionBIOLOGICALACTIVITY?物活性BMS299897?種磺胺類γ-secretase抑制劑，作?于穩(wěn)定過表達淀粉樣前體蛋?(APP)的HEK293細胞，抑制Aβ產(chǎn)?，IC50為7nM。IC50&TargetIC50:7nM(Aβ,inHEK293cells)[1]體外研究BMS-299897reducesthelevelsofeachoftheAβpeptides.At1μM,BMS-299897decreasesthesepeptidestolevelsrangingfrom20to50%ofthevehiclecontrol.BMS-299897treatmentreducestheportionofQD-BDNFsignalsmovingintheretrogradedirection(p=0.0198)withaconcomitantincreaseintheportionofsignalsmovingintheanterogradedirection(p=0.0147)[2].體內(nèi)研究BMS-299897showsdose-andtime-dependentreductionsofamyloidβ-peptide(Aβ)inbrain,cerebrospinalfluid(CSF),andplasmainyoungtransgenicmice,withacorrelationbetweenbrainandCSFAβlevels.BMS-299897reducesbothbrainandplasmaAβ1-40inAPP-YACmiceandincreasesbrainconcentrationsofAPPcarboxy-terminalfragments,consistentwithγ-secretaseinhibition.BMS-299897,attenuatesthisAβ25-35-inducedAβ1-42seedingandtoxicity.BMS-299897isadministeredat0.1-1nmol/mouse,concomittantlywithAβ25-35(9nmol)inmaleSwissmice.Afteroneweek,thecontentsinAβ1-42andAβ1-40,andthelevelsinlipidperoxidationareanalyzedinthemousehippocampus.Micearesubmittedtospontaneousalternation,passiveavoidanceandobjectrecognitiontoanalyzetheirshort-andlong-termmemoryabilities.Aβ25-35increasesAβ1-42content(+240%)butfailstoaffectAβ1-40.BMS-299897blockstheincreaseinAβ1-42contentanddecreasedAβ1-40levelssignificantly.ThecompounddoesnotaffectAβ25-35-inducedincreaseinhippocampallipidperoxidation.Behaviorally,BMS-299897blockstheAβ25-35-induceddeficitsinspontaneousalternationornovelobjectrecognition,usinga1hintertrialtimeinterval.Theco-administrationoftheγ-secretaseinhibitorBMS-299897,inthe0.1-1μmol/mousedose-range,completelyblockstheAβ25-35-inducedincreaseinAβ1-42content[1].PROTOCOLCellAssay[2]NeuronalculturesaretreatedatindicatedDIVsfor24hrswith1μMBMS-299897,2.5μMsGSM41orthevehicleDMSO(finalconcentration:0.1%)formostoftheexperiments,orfordifferentperiodsoftimeasindicatedinspecificexperiments.TransfectionsofsiRNAsareperformedon100,000neuronsatDIV4usingNTERNanoparticletransfectionsystemfollowingtheprotocolprovided.ThesiRNAsusedare:1)siRNAagainstratAPP,and2)theMISSIONsiRNA.KnockdownexperimentsareoptimizedbyexaminingbothmRNAandproteinexpressionofAPP.Aknockdownefficiencyof80%atthemRNAlevelasquantitatedwiththe7300RealTimePCRSystemand30%attheproteinlevelbyimmunoblottingisroutinelyachieved[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1]MaleSwissOF1mice,aged7-9weeksandweighing32±2gareused.Dosesof0.1,0.3and1μmolare2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEinjectedi.c.v.in1μLsimultaneouslywiththeAβ25-35peptide.Animalsareusedatday7-9afteri.c.v.injectionsforbehavioraltestingorsacrifice,beforebiochemicalmeasures.Allexperimentsareconductedonseparatebatchofmice,exceptspontaneousalternationandpassiveavoidance,whicharedoneinseriesinthesameanimals.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].MeunierJ,etal.Theγ-secretaseinhibitor2-[(1R)-1-[(4-chlorophenyl)sulfonyl](2,5-difluorophenyl)amino]ethyl-5-fluorobenzenebutanoicacid(BMS-299897)alleviatesAβ1-42seedingandshort-termmemorydeficitsintheAβ25-35mousemodelofAlzheimer'sd[2].WeissmillerAM,etal.Aγ-secretaseinhibitor,butnotaγ-secretasemodulator,induceddefectsinBDNFaxonaltraffickingandsignaling:evidenceforaroleforAPP.PLoS