Description | BaricitinibisaselectiveorallybioavailableJAK1/JAK2inhibitorwithIC50of5.9nMand5.7nM,respectively. |
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IC50&Target | IC50:5.9nM(JAK1),5.7nM(JAK2),>400(JAK3),53nM(Tyk2)[1] |
InVitro | Incell-basedassays,Baricitinib(INCB028050)provestobeapotentinhibitorofJAKsignalingandfunction.InPBMCs,BaricitinibinhibitsIL-6-stimulatedphosphorylationofthecanonicalsubstrateSTAT3(pSTAT3)andsubsequentproductionofthechemokineMCP-1withIC50valuesof44nMand40nM,respectively.InisolatednaiveT-cells,INCB028050alsoinhibitspSTAT3stimulatedbyIL-23(IC50=20nM).Importantly,thisinhibitionpreventedtheproductionoftwopathogeniccytokines(IL-17andIL-22)producedbyTh17cells-asubtypeofhelperTcellswithdemonstrableinflammatoryandpathogenicproperties-withanIC50valueof50nM.Instarkcontrast,thestructurallysimilarbutineffectiveJAK1/2inhibitorsINCB027753andINCB029843hasnosignificanteffectinanyoftheseassayssystemswhentestedatconcentrationsupto10μM[1]. |
InVivo | Baricitinib(INCB028050)treatment,compareswithvehicle,inhibitstheincreaseinhindpawvolumesduringthe2wkoftreatmentby50%atadoseof1mg/kgand>95%atdosesof3or10mg/kg.Becausebaselinepawvolumemeasurementsaretakenontreatmentday0-inanimalswithsignificantsignsofdisease-itispossibletohave>100%inhibitioninanimalsshowingmarkedimprovementinswelling[1].Baricitinib(0.7mg/day)treatedmiceexhibitssubstantiallyreducedinflammationasassessedbyH&Estaining,reducedCD8infiltration,andreducedMHCclassIandclassIIexpressionwhencomparedwithvehicle-controltreatedmice.CD8+NKG2D+cells,criticaleffectorsofdiseaseinmurineandhumanalopeciaareata(AA),aregreatlydiminishedinBaricitinibtreatedmicecomparewithvehiclecontroltreatedmice[2]. |
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PreparingStockSolutions |
Pleaserefertothesolubilityinformationtoselecttheappropriatesolvent. | ||||||||||||||||
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KinaseAssay [1] | Enzymeassaysareperformedusingahomogeneoustime-resolvedfluorescenceassaywithrecombinantepitopetaggedkinasedomains(JAK1,837-1142;JAK2,828-1132;JAK3,718-1124;Tyk2,873-1187)orfull-lengthenzyme(cMETandChk2)andpeptidesubstrate.Eachenzymereactionisperformedwithorwithouttestcompound(11-pointdilution),JAK,cMET,orChk2enzyme,500nM(100nMforChk2)peptide,ATP(attheKmspecificforeachkinaseor1mM),and2.0%DMSOinassaybuffer.ThecalculatedIC50valueisthecompoundconcentrationrequiredforinhibitionof50%ofthefluorescentsignal.AdditionalkinaseassaysareperformedatCerepusingstandardconditionsat200nM.Enzymestestedincluded:Abl,Akt1,AurA,AurB,CDC2,CDK2,CDK4,CHK2,c-kit,EGFR,EphB4,ERK1,ERK2,FLT-1,HER2,IGF1R,IKKα,IKKβ,JNK1,Lck,MEK1,p38α,p70S6K,PKA,PKCα,Src,andZAP70[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly. | ||||||||||||||||
CellAssay [1] | Baricitinib(INCB028050)isdissolvedinstocksolutions,andthendilutedwithappropriatemediabeforeuse[1]. HumanPBMCsareisolatedbyleukapheresisfollowedbyFicoll-Hypaquecentrifugation.ForthedeterminationofIL-6-inducedMCP-1production,PBMCsareplatedat3.3×105cellsperwellinRPMI1640+10%FCSinthepresenceorabsenceofvariousconcentrationsofINCB028050(1nM,10nM,100nM,1μM,and10μM).Followingpreincubationwithcompoundfor10minatroomtemperature,cellsarestimulatedbyadding10ng/mLhumanrecombinantIL-6toeachwell.Cellsareincubatedfor48hat37°C,5%CO2.SupernatantsareharvestedandanalyzedbyELISAforlevelsofhumanMCP-1.TheABIlityofINCB028050toinhibitIL-6-inducedsecretionofMCP-1isreportedastheconcentrationrequiredfor50%inhibition(IC50).ProliferationofBa/F3-TEL-JAK3cellsisperformedover3dusingCell-TiterGlo[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly. | ||||||||||||||||
AnimalAdmiNISTration [1][2] | Baricitinib(INCB028050)issUSPendedin0.5%methylcellulose(Rat)[1]. Rat[1] | ||||||||||||||||
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MolecularWeight | 371.42 | ||||||||||||
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Formula | C₁₆H₁₇N₇O₂S | ||||||||||||
CASNo. | 1187594-09-7 | ||||||||||||
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Shipping | RoomtemperatureincontinentalUS;mayvaryelsewhere | ||||||||||||
Solvent&Solubility | DMSO:25mg/mL Baricitinibissuspendedin0.5%methylcellulose[3]. *"<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">1> | ||||||||||||
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Purity:99.70%
SouthernBiotech/Goat Anti-Human Lambda-Alexa Fluor® 555/2070-32/1.0 mg
vectorlabs/Biotinylated Aleuria Aurantia Lectin (AAL)/B-1395/1 mg
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Mouse IgG)/PK-6102/1 kit
vectorlabs/VECTASTAIN® Elite® ABC-HRP Kit (Peroxidase, Universal)/PK-6200/1 kit
vectorlabs/Unconjugated Musa Paradisiaca (Banana) Lectin (BanLec)/L-1410/5 mg
GiottoBiotech/Calmodulin N60D/5 mg/G02CLM60cn