CyclosporinAisanimmunosuppressiveagent,thatbindstothecyclophilinandtheninhibitscalcineurinwithIC₅₀of7nMinacell-freeassay,andiswidelyusedinorgantransplantationtopreventrejection.

IC50:7nM(calcineurin)

CyclosporinAbindstothecyclophilin(immunophilin)inTcells^[1],andformsaCyclophilin-CyclosporinAcomplexwhichinhibitscalcineurin^[2]. CyclosporinAinhibitscalcineurinwithIC₅₀of7nM^[3],thenblocksthenucleartranslocationofNF-AT^[4]. CyclosporinAalsoaffectsmitochondriabypreventingtheMTP(mitochondrialpermeABIlitytransitionpore)fromopeningwithanIC₅₀of39nM^[5]. 

CyclosporinAisanimmunosuppressiveagentthatiseffectivefollowingeitherparenteralororaladmiNISTrationinmice,ratandguineapigs^[6]. CyclosporinAisclinicallyusedinorgantransplantationtopreventrejection^[7]. 

• [1].HandschumacherRE,etal.Cyclophilin:aspecificcytosolicbindingproteinforcyclosporinA.Science.1984Nov2;226(4674):544-7.

  [2].LiuJ,etal.Calcineurinisacommontargetofcyclophilin-cyclosporinAandFKBP-FK506complexes.Cell.1991Aug23;66(4):807-15.

  [3].FrumanDA,etal.CalcineurinphosphataseactivityinTlymphocytesisinhibitedbyFK506andcyclosporinA.ProcNatlAcadSciUSA.1992May1;89(9):3686-90.

  [4].FlanaganWM,etal.NuclearassociationofaT-celltranscriptionfactorblockedbyFK-506andcyclosporinA.Nature.1991Aug29;352(6338):803-7.

  [5].NicolliA,etal.Interactionsofcyclophilinwiththemitochondrialinnermembraneandregulationofthepermeabilitytransitionpore,andcyclosporinA-sensitivechannel.JBiolChem.1996Jan26;271(4):2185-92.

  [6].BorelJF,etal.Effectsofthenewanti-lymphocyticpeptidecyclosporinAinanimals.Immunology.1977Jun;32(6):1017-25.

  [7].Williams,R,etal.Randomisedtrialcomparingtacrolimus(FK506)andcyclosporininpreventionofliverallograftrejection.EuropeanFK506MulticentreLiverStudyGroup.Lancet,1994,344(8920),423-428.

Reactionmixtureswithpurifiedenzymecontains100nMcalcineurin,100nMcalmodulin,and5μM ³²P-labeledphosphopeptide,in60μL(totalvolume)ofassaybuffercontaining20mMTris(pH8),100mMNaCl,6mMMgCl₂,0.5mMdithiothreitol,0.1mgofbovineserumalbuminpermL,andeither0.1mMCaCl₂or5mMEGTA.Reactionmixtureswithcelllysatescontains20μLofundilutedlysate,5μM ³²P-labeledphosphopeptide,and40μLofassaybuffer.Reactionmixturescontains50μMpeptide412or413and/or500nMokadaicacid,aspecificinhibitorofphosphatases1and2A;500nMokadaicacidissufficientforinhibitionofCa²⁺-independentphosphatases,whereashigherconcentrationspartiallyinhibitCa²⁺-dependentactivityaswell.After15minat30°C,reactionsareterminatedbytheadditionof0.5mLof100mMpotassiumphosphatebuffer(pH7.0)containing5%trichloroaceticacid.FreeinorganicphosphateisisolatedbyDowexcation-exchangechromatographyandquantitatedbyscintillationcountingasdescribed.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

CyclosporinAisdissolvedinethanol.

Immunosuppressiveagentsaredissolvedinethanolatconcentrations1000-foldmorethantheconcentrationdesiredforcelltreatments.Cells(10⁶)aresUSPendedin1mLofcompletemediuminmicrocentrifugetubes;1μLofethanoloroftheethanolicsolutionofCyclosporinAisadded,andthecellsareincubatedat37°Cfor1hr.Cellsarewashedtwicewith1mLofPBSoniceandlysedin50μLofhypotonicbuffercontaining50mMTris(pH7.5);0.1mMEGTA;1mMEDTA;0.5mMdithiothreitol;and50μgofphenylmethylsulfonylfluoride,50μgofsoybeantrypsininhibitor,5μgofleupeptin,and5μgofaprotininpermL.Lysatesaresubjectedtothreecyclesoffreezinginliquidnitrogenfollowedbythawingat30°Candthenarecentrifugedat4°Cfor10minat12,000×g.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

CyclosporinAissuspendedin0.5%solutionoftragacanth.

Ratsareimmunizedonday0i.p.with05mLandmicei.v.with02mLofa10%suspensionofwashedsheeperythrocytes(SE).Toelicitasecondaryresponse,miceareboosted8-11weeksaftertheprimaryimmunizationwithani.v.injectionof0-2mLof025%washedSE(10⁷cells).Forprolongedtreatmenttheanimalsreceiveontheaverage45mg/kgcyclosporinAdailyinthefoodduring13weeks.Theseratsareimmunized5daysbeforekilling.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

• [1].HandschumacherRE,etal.Cyclophilin:aspecificcytosolicbindingproteinforcyclosporinA.Science.1984Nov2;226(4674):544-7.

  [2].LiuJ,etal.Calcineurinisacommontargetofcyclophilin-cyclosporinAandFKBP-FK506complexes.Cell.1991Aug23;66(4):807-15.

  [3].FrumanDA,etal.CalcineurinphosphataseactivityinTlymphocytesisinhibitedbyFK506andcyclosporinA.ProcNatlAcadSciUSA.1992May1;89(9):3686-90.

  [4].FlanaganWM,etal.NuclearassociationofaT-celltranscriptionfactorblockedbyFK-506andcyclosporinA.Nature.1991Aug29;352(6338):803-7.

  [5].NicolliA,etal.Interactionsofcyclophilinwiththemitochondrialinnermembraneandregulationofthepermeabilitytransitionpore,andcyclosporinA-sensitivechannel.JBiolChem.1996Jan26;271(4):2185-92.

  [6].BorelJF,etal.Effectsofthenewanti-lymphocyticpeptidecyclosporinAinanimals.Immunology.1977Jun;32(6):1017-25.

  [7].Williams,R,etal.Randomisedtrialcomparingtacrolimus(FK506)andcyclosporininpreventionofliverallograftrejection.EuropeanFK506MulticentreLiverStudyGroup.Lancet,1994,344(8920),423-428.

1202.61

C₆₂H₁₁₁N₁₁O₁₂

59865-13-3

RoomtemperatureincontinentalUS;mayvaryelsewhere

10mMinDMSO

*"<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">

Purity:>98.0%

SelectBatch:HY-B0579-11724

DataSheet(124KB)SDS(405KB)

COA(94KB)HNMR(332KB)

HandlingInstructions(1252KB)

• [1].HandschumacherRE,etal.Cyclophilin:aspecificcytosolicbindingproteinforcyclosporinA.Science.1984Nov2;226(4674):544-7.

  [2].LiuJ,etal.Calcineurinisacommontargetofcyclophilin-cyclosporinAandFKBP-FK506complexes.Cell.1991Aug23;66(4):807-15.

  [3].FrumanDA,etal.CalcineurinphosphataseactivityinTlymphocytesisinhibitedbyFK506andcyclosporinA.ProcNatlAcadSciUSA.1992May1;89(9):3686-90.

  [4].FlanaganWM,etal.NuclearassociationofaT-celltranscriptionfactorblockedbyFK-506andcyclosporinA.Nature.1991Aug29;352(6338):803-7.

  [5].NicolliA,etal.Interactionsofcyclophilinwiththemitochondrialinnermembraneandregulationofthepermeabilitytransitionpore,andcyclosporinA-sensitivechannel.JBiolChem.1996Jan26;271(4):2185-92.

  [6].BorelJF,etal.Effectsofthenewanti-lymphocyticpeptidecyclosporinAinanimals.Immunology.1977Jun;32(6):1017-25.

  [7].Williams,R,etal.Randomisedtrialcomparingtacrolimus(FK506)andcyclosporininpreventionofliverallograftrejection.EuropeanFK506MulticentreLiverStudyGroup.Lancet,1994,344(8920),423-428.

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Medchemexpress/Plerixafor octahydrochloride(Synonyms: AMD3100 octahydrochloride; JM3100 octahydrochloride; SID791 octahydroch

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