CustomerValidation
- •JNuclMed.2017May;58(5):762-767.
- •JMedChem.2014Nov13;57(21):9204-10.
- •PLoSOne.2016Nov11;11(11):e0166415.
- •Shivashankar.2014.9.15.
Description | PreladenantisapotentcompetitiveantagonistofthehumanA2Areceptor(Ki=1.1nM)andhas>1000-foldselectivityoverallotheradenosinereceptors. |
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IC50&Target | Ki:1.1nM(AdenosineA2Areceptor)[1] |
InVitro | PreladenantalsocompletelyantagonizescAMPincellsexpressingtherecombinanthumanA2Areceptor.PreladenantisdeterminedtohasKBvaluesof1.3nMattheA2Areceptor;thevalueisingoodagreementwiththeKivaluedeterminedintherADIoligandbindingassay.AsimilarfunctionalassaywithA2Breceptor-expressingcellsisusedtodemonstrateselectivityoverA2Breceptors.Inthisassay,theKBvalueforPreladenantis1.2μM,indicatingthatPreladenantis923-foldselectivefortheA2AreceptorovertheA2Breceptor[1]. |
InVivo | Preladenant(1mg/kg)inhibitsL-Dopa-inducedbehavioralsensitizationafterrepeateddailyadministration,whichsuggestsareducedriskofthedevelopmentofdyskinesias.Preladenantexhibitsantidepressant-likeprofilesinmodelsofbehavioraldespair,namelythemousetailsUSPensiontestandthemouseandratforcedswimtest[1].Preladenantproducesadose-dependentreductioninparkinsonianscoresatdosesof1mg/kg(minscore:9.0)and3mg/kg(minscore:6.5).AsubthresholddoseofPreladenantreducesminimumandmeanparkinsonianscoresinanimalstreatedwith3mgkgofL-Dopato5.25and6.88respectively.AWilcoxintestisusedtocompareindividualtreatmentsagainstvehicle.Preladenant(3mg/kg),L-Dopa(3,6,and12mg/kg),andthecombinationofPreladenantandL-Dopa(1or3mg/kg+3mg/kg)areallsignificantlyimprovedontheminimumparkinsonianscore.Inaddition,boththe12mg/kgL-DopaandL-Dopa+Preladenantgroupsaresignificantlyimprovedonbothminimumandmeanparkinsonianscoresrelativetothe3mg/kgL-Dopagroup[2]. |
ClinicalTrial | ViewMoreCollapse |
References |
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Pleaserefertothesolubilityinformationtoselecttheappropriatesolvent. | ||||||||||||||||
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KinaseAssay [1] | Receptorbindingisperformedusingmembranespreparedfromcellswithrecombinantexpressionofadenosinereceptorsasfollows:humanA2AandHEK293,ratA2AandChinesehamsterovary,humanandratA1andChinesehamsterovary,andhumanA3andHEK293.Radioligandcompetitionbindingassaysareperformedin96-wellplatesinatotalassayvolumeof200μLusingafinaltestdrugconcentrationrangeofbetween0.1and3μM.Membranesaredilutedinassaybuffer,pH7.4(A1andA2A,Dulbecco"sphosphate-bufferedsalinewith10mMMgCl2;A3,50mMTris-HCl,120mMNaCl,10mMMgCl2).Toremoveendogenousadenosinefromthemembranepreparations,4U/mLadenosinedeaminaseisaddedtothemembranes,whicharethenincubatedatroomtemperaturefor15min.Radioligandisaddedtoafinalconcentrationof0.5([3H]SCH58261,A2A),1([3H]DPCPX,A1),or0.25([125I]AB-MECA,A3)nM.Nonspecificbindingisdefinedbyadding100nMCGS15923(A2A),100nMNECA(A1),or100nMDPCPX(A3).Platesareincubatedatroomtemperaturewithagitationfor1.5h(A2AandA1)or2h(A3).MembranesarefilteredontoPackardGF-Bfilterplatesandwashedinice-coldassaybufferusingaBrandelcellharvestertoseparateboundandfreeradioligand.Theplatesaredriedbeforeadditionof45μLofMicroscint20toeachwell.IC50valuesaredeterminingbyfittingthedisplacementcurvesusinganiterativecurve-fittingprogram.KivaluesarecalculatedusingtheCheng-Prusoffequation[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly. | ||||||||||||||||
CellAssay [1] | PreladenantisdissolvedinDMSOandstored,andthendilutedwithappropriatemediabeforeuse[1]. HEK293cellsstablyexpressingeitherhumanA2AorA2Breceptorsaregrowntoconfluence,harvestedusingenzyme-freecelldissociationbufferandpelletedbycentrifugation(1000g;5min).Thecellsarewashedanddilutedtoafinaldensityof4×106cells/mLinHanks"balancedsaltsolutionsupplementedwith10nMHPS,pH7.4,,5mMMgCl2,and0.2%bovineserumalbumin.Preladenantisdilutedintheabovebufferwithinclusionofthefollowingcomponentstoachievetherespectivefinalassayconcentrations:0.25%DMSO,2U/mLadenosinedeaminase,and100μMRo201724.Cellsuspensions(20μL)arepreincubatedfor15minatroomtemperaturein96-wellplatescontaining25μLofvehicleorPreladenant.CGS-21680(A2A)or5-N-cyclopropylcarboxamidoadenosine(A2B)at10-foldthedesiredconcentrationisthenadded,andthereactionsareincubatedfor15minat37°C.Thereactionsareterminatedbytheadditionof50μLofassay/lysisbuffer.TheconcentrationresponsecurvesforCGS-21680inthepresenceandabsenceofPreladenantareplotted,andtheEC50valuesaredeterminedbyfittingthecurvesusingGraphPadPrismsoftware[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly. | ||||||||||||||||
AnimalAdministration [1][2] | Preladenantispreparedin50%polyethyleneglycol400(RatandMice)[1]. MiceandRat[1] | ||||||||||||||||
References |
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MolecularWeight | 503.56 | ||||||||||||
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Formula | C₂₅H₂₉N₉O₃ | ||||||||||||
CASNo. | 377727-87-2 | ||||||||||||
Storage |
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Shipping | RoomtemperatureincontinentalUS;mayvaryelsewhere | ||||||||||||
Solvent&Solubility | DMSO *"<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">1> Purity:99.08%
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