Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEGI254023XCat.No.:HY-19956CASNo.:260264-93-5Synonyms:GI4023;SRI028594分?式:C??H??N?O?分?量:391.5作?靶點(diǎn):MMP作?通路:MetabolicEnzyme/Protease儲(chǔ)存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:100mg/mL(255.43mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM2.5543mL12.7714mL25.5428mL5mM0.5109mL2.5543mL5.1086mL10mM0.2554mL1.2771mL2.5543mL請(qǐng)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(chǔ)備液；?旦配成溶液，請(qǐng)分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲(chǔ)備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?，-20°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?。體內(nèi)實(shí)驗(yàn)請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥?式選擇適當(dāng)?shù)娜芙?案。以下溶解?案都請(qǐng)先按照InVitro?式配制澄的儲(chǔ)備液，再依次添加助溶劑：(為保證實(shí)驗(yàn)結(jié)果的可靠性，澄的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)保存；體內(nèi)實(shí)驗(yàn)的?作液，建議您現(xiàn)?現(xiàn)配，當(dāng)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(qǐng)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(6.39mM);Clearsolution2.請(qǐng)依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(6.39mM);Clearsolution3.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(6.39mM);ClearsolutionBIOLOGICALACTIVITY?物活性GI254023X?種有效的MMP9和ADAM10抑制劑,IC50s分別為2.5和5.3nM。IC50&TargetIC50:2.5nM(MMP9),5.3nM(ADAM10)[1]體外研究Incellularassay25μMandevenaconcentrationof1μMGI254023XstronglyreducesconstitutiveRAGEshedding;alsoPACAP-inducingsheddingofRAGEissignificantlyreduced.Ataconcentrationof100nM,aslightinhibitionofRAGEsheddingisstillobserved.Ininvitroassayswithrecombinantproteinases,GI254023XdiscriminatesbetweenADAM17(IC50=541nM)andADAM10(IC50=5.3nM)/MMP9(IC50=2.5nM)[1].CXCL16sheddingisinhibitedbyADAMproteaseinhibitors(e.gGI254023x).A2780cellsareincubatedwiththeADAM-10/ADAM-17inhibitorTAPI-2,aswellastheADAM-10-selectiveinhibitorGI254023x,asthelevelofexpressedADAM-10isonaverage9.8-foldhigheronmRNAlevelcomparewithADAM-17.Inaddition,GI254023xalsopreventsCXCL16sheddingfromthecellmembraneandisevenmorepotentthanTAPI-2[2].WhenapplythespecificADAM10(α-secretase)inhibitorGI254023X(5mM)toserum/glucose-deprivedslices,PIcountsaresignificantlyincreasedincomparisonwithDMSO(carrier)-treatedcontrols[3].PROTOCOLCellAssay[3]Celldeathisquantifiedbasedonplasmamembranepermeabilization.WhenapplytheADAM10(a-secretase)inhibitorGI254023X(5mM),slicesareculturedinserum-/glucose-freemediumfor48hcontainingtheinhibitororitsrespectivecarrier(DMSO)ascontrol.Roundcirclesofidenticalsize(?500mm)arepositionedinequivalentlocationswithintheCA1regionofeachhippocampusimageandallPI-stainedcellsarecountedusingsoftware.Cellviabilityassaysareperformedwithacommercialkitaccordingtothemanufacturer’sinstructions.TheassayquantitatesATPlevels,anindicatorofmetabolicallyactivecells,photometricallywithafluorescenceplatereader.Additionally,thelive-deadcellstainingkitareappliedaccordingtothemanual.Cellsaresimultaneouslystainedwithgreenfluorescentcalcein-AM(4mM;ex/em:495/515nm)todetectintracellularesteraseactivity(viablecells)andredfluorescentethidiumhomodimer-3(2mM;ex/em:530/635nm)toindicatelossofplasmamembraneintegrity(deadcells)[3].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?Elife.2021Sep20;10:e67261.?GenesDis.2020Nov21;8(6):867-881.2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE?Traffic.2022Nov22.?FrontAgingNeurosci.2021Apr15;13:660249.?MediatInflamm.2019Oct31;2019:5306541.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].VerenaV.Metz,etal.InductionofRAGESheddingbyActivationofGProtein-CoupledReceptors.PLoSOne.2012.[2].MJMGooden,etal.ElevatedserumCXCL16isanindependentpredictorofpoorsurvivalinovariancancerandmayreflectpro-metastaticADAMproteaseactivity.BritishJournalofCancer(2014)110,1535–1544.[3].NMilosch,etal.Holo-APPandG-protein-mediatedsignalingarerequiredforsAPPa-inducedactivationoftheAkt.CellDeathDis.2014Aug28;5:e1