Description | PCI-32765isaselective,irreversibleBtkinhibitorwithIC50valueof0.5nM. |
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IC50&Target | IC50:0.5nM(Btk) |
InVitro | PCI-32765selectivelyinhibitsB-cellsignalingandactivation.ItinhibitsautophosphorylationofBtk(IC50=11nM),phosphorylationofBtk"sphysiologicalsubstratePLCγ(IC50=29nM),andphosphorylationofafurtherdownstreamkinase,ERK(IC50=13nM)[1].PCI-32765inhibitsBCR-activatedprimaryBcellproliferation(IC50=8nM).FollowingFcγRstimulation,PCI-32765inhibitsTNFα,IL-1βandIL-6productioninprimarymonocytes(IC50=2.6,0.5,3.9nM,respectively)[3]. |
InVivo | PCI-32765(3.125-50mg/kg,p.o.)reducesthelevelofcirculatingautoantibodiesandcompletelysuppressesdiseaseinmicewithcollagen-inducedarthritis.PCI-32765inhibitsautoantibodyproductionandthedevelopmentofkidneydiseaseintheMRL-Fas(lpr)lupusmodel.PCI-32765(3.125-50mg/kg,p.o.)reducesrenaldiseaseandautoantibodyproductioninMRL-Fas(lpr)mice[1].PCI-32765(0.1μM)inhibitsactivation-inducedproliferationofCLLcells,inducesselectivecytotoxicityinBcellscomparedwithTcells,butaltersactivationinducedT-cellcytokineproduction[2].PCI-32765dose-dependentlyandpotentlyreversesarthriticinflammationinatherapeuticCIAmodelwithanED50of2.6mg/kg/day.PCI-32765alsopreventsclinicalarthritisinCAIAmodels[3]. |
ClinicalTrial | ViewMoreCollapse |
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Pleaserefertothesolubilityinformationtoselecttheappropriatesolvent. | ||||||||||||||||
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CellAssay [3] | PCI-32765isdissolvedinDMSO. PrimaryhumanBcellsareisolatedfromperipheralbloodmononuclearcellofhealthyhumanvolunteersbyFicoll-HypaquegrADIentseparationfollowedbynegativeselectionusinghumanMiltenylhumanBcellIsolationKitII.In0.2mLRPMIplus10%FBS,100,000BcellsaretreatedwithPCI-32765(0.3nM-10μM)intriplicatewellsorvehiclecontrolin0.1%DMSOfinalconcentrationfor30minutesat37°C,5%CO2,thencellsarestimulatedwith10μg/mLanti-IgMF(ab")2,5μg/mLanti-CD3/CD28asanegativecontrolor0.5μg/mLPMA(Phorbal12-myristate13-acetate)asapositivecontrol.Bcellsarestimulatedfor72hoursat37°C,5%CO2.ProliferationismeasuredwithCellTiterGloreagentandmeasuredonaluminometer.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly. | ||||||||||||||||
AnimalAdmiNISTration [3] | CI-32765isformulatedin1%methylcellulose,0.4%Cremephor®EL,and98.09%water. MaleDBA1/1OlaHsdmiceareinjectedondays0and21withFreunds"CompleteAdjuvantcontainingbovinetypeIIcollagen.Ondays21to35,micearerandomizedintotreatmentgroupswhentheaverageclinicalscoreofeachanimalis1.5(inascaleof5).PCI-32765treatment(1.56-12.5mg/kg,p.o.)isinitiatedfollowingenrollmentandcontinuesfor18days.Clinicalscoresaregiventoeachmousedailyforeachpaw.Clinicalscoreassessmentismadeusingthefollowingcriteria:0=normal;1=onehindpaworforepawjointaffectedorminimaldiffuseerythemaandswelling;2=twohindorforepawjointsaffectedormilddiffuseerythemaandswelling;3=threehindorforepawjointsaffectedormoderatediffuseerythemaandswelling;4=markeddiffuseerythemaandswellingorfourdigitjointsaffected;5=severediffuseerythemaandsevereswellingofentirepaw,unabletoflexdigits.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly. | ||||||||||||||||
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MolecularWeight | 440.5 |
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Formula | C₂₅H₂₄N₆O₂ |
CASNo. | 936563-96-1 |
Storage | 4°C,protectfromlight |
Shipping | RoomtemperatureincontinentalUS;mayvaryelsewhere |
Solvent&Solubility | DMSO:≥52mg/mL PCI-32765isdissolvedinDMSOandthensUSPendedinnormalsaline(0.9%NaCl,NS)[4]. *"<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">1> |
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Purity:99.87%ee.:99.50%
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