Hotline:400-820-3792Inhibitors?Agonists?ScreeningLibrarieswww.MedChemEResiquimodCat.No.:HY-13740CASNo.:144875-48-9分?式:C??H??N?O?分?量:314.38作?靶點(diǎn):Toll-likeReceptor(TLR);HCV作?通路:Immunology/Inflammation;Anti-infection儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMF:50mg/mL(159.04mM;Needultrasonic)掃描?維碼，DMSO:≥30mg/mL(95.43mM)運(yùn)?溶解?案計(jì)算器Methanol:25mg/mL(79.52mM;Needultrasonic)獲得適合您實(shí)驗(yàn)體系的溶解?案H2O:<0.1mg/mL(insoluble)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM3.1809mL15.9043mL31.8086mL5mM0.6362mL3.1809mL6.3617mL10mM0.3181mL1.5904mL3.1809mL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，并請注意儲備液的保存?式和期限。體內(nèi)實(shí)驗(yàn)請根據(jù)您的實(shí)驗(yàn)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(shí)驗(yàn)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(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.請依序添加每種溶劑：10%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.08mg/mL(6.62mM);Clearsolution此?案可獲得≥2.08mg/mL(6.62mM，飽和度未知)的澄溶液。1/4www.MedChemEwww.MedChemE以1mL?作液為例，取100μL20.8mg/mL的澄DMSO儲備液加到400μLPEG300中，混合均勻；向上述2.體系中加?50μLTween-80，混合均勻；然后繼續(xù)加?450μL?理鹽?定容?1mL。請依序添加每種溶劑：10%DMSO90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(6.62mM);Clearsolution此?案可獲得≥2.08mg/mL(6.62mM，飽和度未知)的澄溶液。以1mL?作液為例，取100μL20.8mg/mL的澄DMSO儲備液加到900μL20%的SBE-β-CD?理鹽??溶3.液中，混合均勻。請依序添加每種溶劑：10%DMSO90%cornoilSolubility:≥2.08mg/mL(6.62mM);Clearsolution此?案可獲得≥2.08mg/mL(6.62mM，飽和度未知)的澄溶液，此?案不適?于實(shí)驗(yàn)周期在半個?以上的實(shí)驗(yàn)。4.以1mL?作液為例，取100μL20.8mg/mL的澄DMSO儲備液加到900μL??油中，混合均勻。請依序添加每種溶劑：5%DMSO40%PEG3005%Tween-8050%salineSolubility:≥2.5mg/mL(7.95mM);ClearsolutionBIOLOGICALACTIVITY?物活性Resiquimod?種Toll樣受體7和8(TLR7/TLR8)的激動劑，誘導(dǎo)細(xì)胞因?如TNF-α，IL-6和IFN-α的上調(diào)。IC50&TargetTLR7/TLR8體外研究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].體內(nèi)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.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?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