NIK-SMI1-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemENIKSMI1Cat.No.:HY-112433CASNo.:1660114-31-7分⼦式:C₂₀H₁₉N₃O₄分⼦量:365.38作⽤靶点:NF-κB作⽤通路:NF-κB储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:125mg/mL(342.11mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.7369mL13.6844mL27.3688mL5mM0.5474mL2.7369mL5.4738mL10mM0.2737mL1.3684mL2.7369mL请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；⼀旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存⽅式和期限：-80°C,6months;-20°C,1month。-80°C储存时，请在6个⽉内使⽤，-20°C储存时，请在1个⽉内使⽤。体内实验请根据您的实验动物和给药⽅式选择适当的溶解⽅案。以下溶解⽅案都请先按照InVitro⽅式配制澄的储备液，再依次添加助溶剂：(为保证实验结果的可靠性，澄的储备液可以根据储存条件，适当保存；体内实验的⼯作液，建议您现⽤现配，当天使⽤；以下溶剂前显⽰的百分⽐指该溶剂在您配制终溶液中的体积占⽐；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的⽅式助溶)1.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.08mg/mL(5.69mM);Clearsolution1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(5.69mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(5.69mM);Clearsolution4.请依序添加每种溶剂：5%DMSO>>40%PEG300>>5%Tween-80>>50%salineSolubility:≥2.5mg/mL(6.84mM);Clearsolution5.请依序添加每种溶剂：5%DMSO>>95%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(6.84mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性NIKSMI1⼀种有效的选择性NF-κB诱导激酶(NIK)抑制剂，可抑制NIK催化的ATP⽔解为ADP，IC50为0.23±0.17nM。IC50&TargetNIK[1]体外研究NIKSMI1(Compound4f)inhibitsNIK-catalyzedhydrolysisofATPtoADP(fluorescencepolarization,FP)withanIC50of0.23±0.17nM.NIKSMI1inhibitstheexpressionofNIKSMI1responseelementregulatedfireflyluciferasereportergeneinHEK293cellswithanIC50of34±6nM.ConsistentwithexpectationsforaNIKinhibitor,NIKSMI1isshowntoinhibitnucleartranslocationofp52(RelB)(IC50=70nM).NIKSMI1inhibitsBAFF-inducedBcell(mouse)survivalinvitrowithanIC50of373±64nM[1].体内研究C57BL/6micearetreatedtwicedailyfor7dayswithorallyadministeredNIKSMI1orwiththreeinjectionsofrecombinantBAFFreceptorfusionprotein(Br3-mIgG2a)overthecourseofthe7-dayexperimentasapositivecontrol.Thenonlinearityofexposurerelativetodosebetween100and200mg/kgisaresultofsaturationofclearancemechanisms.ThepharmacologyofNIKSMI1isexaminedinSDrat,CD-1mouse,beagle,andcynomologousmonkeywith20,32,18,and7.8mL/kgpermin,respectively.Volumeofdistribution(Vd,L/kg)is1.35,1.58,0.778,and1.39,respectively[1].PROTOCOLCellAssay[1]HumanBcellsarere-suspendedinRPMIwith10%FBSfortheproliferationassaysand2.5%FBSforthesurvivalassays.MouseBcellsareplatedinCo-star96-wellplatesateither50,000cells/wellforthesurvivalassaysorat150,000cells/wellfortheproliferationassays.Compounds(e.g.,NIKSMI1)dilutedinDMSO(finalDMSOassayconcentration=0.1%)areaddedtothecells.ThecellsareincubatedwithNIKSMI1foronehourat37°C.Stimulusisthenaddedtotheplatesandsurvivalorproliferationismeasuredafterfourdays.Fortheproliferationassays,cellsaretreatedwitheitherAnti-IgM(20ꢀµg/mL)orrhCD40L(10ꢀµg/mL)oranti-mouseCD40(100ꢀng/mL).FortheBAFFsurvivalassay,cellsaretreatedwithhumanormouserBAFFat10ꢀng/mLfollowedbyCellTiterGlotomeasuresurvivalondayfour[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEAdministration[1]Age-matchedC57BL/6miceareused.Onlyfemalemiceareusedintheseexperiments.ThesingleoraldosesofNIKSMI1are10,20,60,100,and200mg/kg.ForPOdosing,animalsaremanuallyrestrained,thendosedviaoralgavageusinganappropriatelysizedgavageneedle.Animalsaremonitoredforanysignsofaspirationordistress-respiratoryabnormalities,lethargy,paleextremities,etc.Forsamplecollection,3micepergrouparebledatotalof8timesviatailprickusinga27Gneedle(lateraltailvein).10μLofbloodiscollectedateachtimepointanddepositedintoapre-filledcostarclustertubecontaining40μLof1.7mg/mLEDTA/water,thetubeiscapped,votexedfor5seconds,thenstoredondryice.Samplesaretransferredtoa-80°Cfreezerforstorage[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•NatImmunol.2020May;21(5):535-545.•SciImmunol.2022Aug12;7(74):eabn3800.•MolNeurobiol.2021Jan13.•JImmunolRes.2020Jul31;2020:1859260.•ResearchSquarePrint.2022Jun.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].BlaquiereN,etal.Scaffold-HoppingApproachToDiscoverPotent,Selective,andEfficaciousInhibitorsofNF-κBInducingKinase.JMed