Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemENIKSMI1Cat.No.:HY-112433CASNo.:1660114-31-7分?式:C??H??N?O?分?量:365.38作?靶點:NF-κB作?通路:NF-κB儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:125mg/mL(342.11mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.7369mL13.6844mL27.3688mL5mM0.5474mL2.7369mL5.4738mL10mM0.2737mL1.3684mL2.7369mL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(fù)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(dāng)?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(dāng)保存；體內(nèi)實驗的?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.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.08mg/mL(5.69mM);Clearsolution1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(5.69mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(5.69mM);Clearsolution4.請依序添加每種溶劑：5%DMSO>>40%PEG300>>5%Tween-80>>50%salineSolubility:≥2.5mg/mL(6.84mM);Clearsolution5.請依序添加每種溶劑：5%DMSO>>95%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(6.84mM);ClearsolutionBIOLOGICALACTIVITY?物活性NIKSMI1?種有效的選擇性NF-κB誘導(dǎo)激酶(NIK)抑制劑，可抑制NIK催化的ATP?解為ADP，IC50為0.23±0.17nM。IC50&TargetNIK[1]體外研究NIKSMI1(Compound4f)inhibitsNIK-catalyzedhydrolysisofATPtoADP(fluorescencepolarization,FP)withanIC50of0.23±0.17nM.NIKSMI1inhibitstheexpressionofNIKSMI1responseelementregulatedfireflyluciferasereportergeneinHEK293cellswithanIC50of34±6nM.ConsistentwithexpectationsforaNIKinhibitor,NIKSMI1isshowntoinhibitnucleartranslocationofp52(RelB)(IC50=70nM).NIKSMI1inhibitsBAFF-inducedBcell(mouse)survivalinvitrowithanIC50of373±64nM[1].體內(nèi)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(20?μg/mL)orrhCD40L(10?μg/mL)oranti-mouseCD40(100?ng/mL).FortheBAFFsurvivalassay,cellsaretreatedwithhumanormouserBAFFat10?ng/mLfollowedbyCellTiterGlotomeasuresurvivalondayfour[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]2/3MasterofBioactiveMolecules—您?邊的抑制劑?師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.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?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