Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEMD2-IN-1Cat.No.:HY-103483CASNo.:111797-22-9分?式:C??H??O?分?量:358.39作?靶點(diǎn):Toll-likeReceptor(TLR)作?通路:Immunology/Inflammation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:50mg/mL(139.51mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM2.7903mL13.9513mL27.9026mL5mM0.5581mL2.7903mL5.5805mL10mM0.2790mL1.3951mL2.7903mL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(guò)程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過(guò)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%salineSolubility:≥3.25mg/mL(9.07mM);Clearsolution1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE2.請(qǐng)依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:3.25mg/mL(9.07mM);Suspendedsolution;Needultrasonic3.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥3.25mg/mL(9.07mM);ClearsolutionBIOLOGICALACTIVITY?物活性MD2-IN-1髓樣分化蛋?2(MD2)的抑制劑，對(duì)重組?MD2(rhMD2)的KD值為189μM。IC50&TargetKD:189μM(rhMD2)[1]體外研究Myeloiddifferentiationprotein2(MD2)isaco-receptorofTLR4.Amongthosederivatives,MD2-IN-1(compound20)showsthestrongestinhibitoryeffectonLPS-inducedexpressionofbothTNF-αandIL-6.Comparetothevehicle,LPSalonelargelyincreasestheamountofTLR4/MD2complex,whilepretreatmentwithMD2-IN-1inhibitstheincreaseofTLR4/MD2complextothevehiclelevel.SPRanalysisshowsthatMD2-IN-1exhibitsrecognizablebindingtorhMD2proteininadose-dependentmanner,withaKDvalueof189?μM,whiletheKDvalueofxanthohumolbindingtoMD2is460?μM.Pre-treatmentwithdifferentdosesofMD2-IN-1dose-dependentlyreducesFITC-LPSbindingtoMD2incellsurfacemembranes,witha65%inhibitionat10?μMintermsofmeanfluorescenceintensity.PretreatmentwithMD2-IN-1alsodose-dependentlyblocksLPS-inducedMAPKphosphorylationintheMPMs[1].體內(nèi)研究AdministrationofMD2-IN-1evidentlyreducestheLPS-inducedincreaseinproteinconcentrationsinBALF.Thelungwet/dryweightratioismarkedlyhigherintheLPS-treatedgroupthanthecontrolgroup,andMD2-IN-1treatmentreducesLPS-inducedpulmonaryedema.LPSalsocausesobservablelunghistopathologicchanges,includingareasofinflammatoryinfiltration,hemorrhage,interstitialedema,thickeningofthealveolarwall,andlungtissuedestruction.ThesehistopathologicalchangesareamelioratedintheMD2-IN-1treatmentgroup[1].PROTOCOLCellAssay[1]MouseRAW264.7macrophagesarestarvedfor3?hbeforeexperimentation.CellsareincubatedwithorwithoutFITC-LPS(50?μg/mL)inthepresenceorabsenceofMD2-IN-1(0.1,1and10?μM)for30?min.Afterincubation,macrophagesarefixedwithparaformaldehydefor10?minat4°CandwashedwithPBSbeforebeinganalyzedbyflowcytometry[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMaleSpragueDawley(SD)ratsarerandomlydividedintothreegroups,designated“control”(5rats,onlyAdministration[1]receivethevehicleof0.9%saline),“LPS”(7rats,receive5?mg/kgLPSalone)and“MD2-IN-1(20)?+?LPS”(6rats,receivebothMD2-IN-1and5?mg/kgLPS).PriortoLPS-inducedAcutelunginjury(ALI),theMD2-IN-1+LPSgroupratsaretreatedintragastricallywithMD2-IN-1atadosageof20?mg/kg/daycontinuouslyforoneweek.Underetheranesthesia,alltheratsareexposedtheirtracheaandchallengedwithintratrachealinstillationof50?μLofLPS,whilethecontrolgroupchallengedwithintratrachealinstillationof50?μLof0.9%2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEsaline.Ratsaretheneuthanizedwithketamineafter6?hofLPSinduction[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].ZhangY,etal.DiscoveryofnewMD2inhibitorfromchalconederivativeswithanti-inflammatoryeffectsinLPS-inducedacutelunginjury.SciRep.2016Apr27;6:251