Resiquimod-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•Agonists•ScreeningLibrarieswww.MedChemEResiquimodCat.No.:HY-13740CASNo.:144875-48-9分⼦式:C₁₇H₂₂N₄O₂分⼦量:314.38作⽤靶点:Toll-likeReceptor(TLR);HCV作⽤通路:Immunology/Inflammation;Anti-infection储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMF:50mg/mL(159.04mM;Needultrasonic)扫描⼆维码，DMSO:≥30mg/mL(95.43mM)运⽤溶解⽅案计算器Methanol:25mg/mL(79.52mM;Needultrasonic)获得适合您实验体系的溶解⽅案H2O:<0.1mg/mL(insoluble)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制备储备液1mM3.1809mL15.9043mL31.8086mL5mM0.6362mL3.1809mL6.3617mL10mM0.3181mL1.5904mL3.1809mL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，并请注意储备液的保存⽅式和期限。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%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.08mg/mL(6.62mM);Clearsolution此⽅案可获得≥2.08mg/mL(6.62mM，饱和度未知)的澄溶液。1/4www.MedChemEwww.MedChemE以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到400μLPEG300中，混合均匀；向上述2.体系中加⼊50μLTween-80，混合均匀；然后继续加⼊450μL⽣理盐⽔定容⾄1mL。请依序添加每种溶剂：10%DMSO90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(6.62mM);Clearsolution此⽅案可获得≥2.08mg/mL(6.62mM，饱和度未知)的澄溶液。以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到900μL20%的SBE-β-CD⽣理盐⽔⽔溶3.液中，混合均匀。请依序添加每种溶剂：10%DMSO90%cornoilSolubility:≥2.08mg/mL(6.62mM);Clearsolution此⽅案可获得≥2.08mg/mL(6.62mM，饱和度未知)的澄溶液，此⽅案不适⽤于实验周期在半个⽉以上的实验。4.以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到900μL⽟⽶油中，混合均匀。请依序添加每种溶剂：5%DMSO40%PEG3005%Tween-8050%salineSolubility:≥2.5mg/mL(7.95mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性Resiquimod⼀种Toll样受体7和8(TLR7/TLR8)的激动剂，诱导细胞因⼦如TNF-α，IL-6和IFN-α的上调。IC50&TargetTLR7/TLR8体外研究Resiquimod(R-848)inducesbothhapten-andallergen-specificcirculatingTcells,includingTH2effectors,toproduceIFN-γandeventolosetheabilitytoproduceIL-4[2].Resiquimod(R848)enhancesPBLproliferationinadose-dependentmanner,andincreasesthenumberofBrdU-positivecellsinBrdUincorporationassay.CellstreatedwithR848exhibitssignificantlyincreased(3.5-fold)luciferase(areporterofNF-κBactivity)activity[3].体内研究Resiquimod(R-848)(50μg/bird,i.m.route)significantlyup-regulatestheexpressionofIFN-α,IFN-β,IFN-γ,IL-1β,IL-4,iNOSandMHC-IIgenesinSPFchicken[1].PROTOCOLKinaseAssay[3]Forluciferaseassay,FG-9307cellsaretransfectedwiththefireflyNF-κB-specificluciferasereportervectorpNFκB-Met-Luc2.Transfectionefficiencyismonitoredbyco-transfectionwiththepSEAP2controlvector,whichconstitutivelyexpressesthehumansecretedenhancedalkalinephosphatase(SEAP).ThenthecellsaretreatedwithResiquimod(R848,1µg/mL),CQ(10µM),CQplusR848orPBSandincubatedat22°Cfor24h.TheculturemediumofthetransfectantsisthenanalyzedforluciferaseactivityandSEAPactivityusingLuciferaseAssayKitandtheGreatEscAPe™SEAPChemiluminescenceDetectionKit,respectively.Theassayisperformedthreetimes.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[3]Forinhibitionoflysosomalacidification,cellsareincubatedwith10µMCQfor1hbeforeResiquimod(R848)treatment.Aftertreatment,20µLof5mg/mLMTTisaddedtotheplate.Theplateisincubatedat22°Cfor4h,and200µLdimethylsulfoxideisaddedtotheplatetodissolvethereducedformazan.Theplateisthen2/4www.MedChemEwww.MedChemEreadat490nmwithamicroplatereader.TodeterminetheeffectofMyd88inhibitiononR848-inducedcellproliferation,theMyd88inhibitorPepinh-MYDandthecontrolpeptidePepinh-ControlareaddedtoPBLattheconcentrationof50µM,andtheplateisincubatedat22°Cfor6h.Afterincubation,thecellsaretreatedwithR848andsubjectedtoMTTassayasabove.TodeterminetheeffectofNF-κBinactivationonR848-inducedcellproliferation,BAY-11-7082,anirreversibleinhibitorofIκB-αphosphorylation,isaddedtothecellsattheconcentrationof1µM,andtheplateisincubatedat22°Cfor1h.Afterincubation,thecellsaretreatedwithR848andsubjectedtoMTTassayasearlier.Allexperimentsareperformedthreetimes.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalAtotalof40SPFchickensoftwo-weekoldareallottedtooneofthefollowingfourexperimentalgroupsAdministration[1](n=10/group):GroupA:PBScontrol;GroupB:inactivatedNDVvaccine;GroupC:commercialoiladjuvantedinactivatedNDVvaccinepreparedfromlentogenicstrainandGroupD:combinationofinactivatedNDVvaccineandR-848(50μg/bird).VaccineorPBSisadministeredbyintramuscularrouteinthethighmuscle.Aboosterdoseisgiven14-daypostimmunization(d.p.i).Twoweekspost-booster,experimentalSPFbirdsarechallengedwithvelogenicstrainofNDV(105ELD50perbird)intramuscularly.Clinicalsignsandmortalityareobserveddailytill14daypost-challenge(d.p.c).Cloacalswabs(n=6/group)arecollectedfromthebirdsonday0,4,7and14post-challengeandinoculatedinto10-dayoldembryonatedchickeneggs(n=3eggs/sample)throughintra-allantoicroute.Threedaypost-inoculation,theallantoicfluidischeckedfortheNDVgrowthbyspothaemagglutinationusing10%chickenRBC.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•NatBiomedEng.2018Aug;2(8):578-588.•Small.2020Dec;16(50):e2004905.•ArthritisRheumatol.2020Jan;72(1):166-178.•ActaPharmSinB.2020June29.•IntJNanomedicine.2019Aug30;14:7053-7064.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].SachanS,etal.AdjuvantpotentialofresiquimodwithinactivatedNewcastlediseasevaccineanditsmechanismofactioninchicken.Vaccine.2015Aug26;33(36):4526-32.[2].BrugnoloF,etal.ThenovelsyntheticimmuneresponsemodifierR-848(Resiquimod)shiftshumanaller