Decernotinib-VX-509-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEDecernotinibCat.No.:HY-12469CASNo.:944842-54-0Synonyms:VX-509;VRT-831509分?式:C??H??F?N?O分?量:392.38作?靶點(diǎn):JAK作?通路:Epigenetics;JAK/STATSignaling;ProteinTyrosineKinase/RTK;StemCell/Wnt儲(chǔ)存?式:4°C,protectfromlight,storedundernitrogen*Insolvent:-80°C,6months;-20°C,1month(protectfrom

light,storedundernitrogen)溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:≥50mg/mL(127.43mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM2.5485mL12.7427mL25.4855mL5mM0.5097mL2.5485mL5.0971mL10mM0.2549mL1.2743mL2.5485mL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(protectfromlight,storedundernitrogen)。-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)1/4MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE1.請(qǐng)依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(6.37mM);Clearsolution2.請(qǐng)依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(6.37mM);Clearsolution3.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(6.37mM);ClearsolutionBIOLOGICALACTIVITY?物活性Decernotinib?種有效的，可?服的JAK3抑制劑，對(duì)JAK3，JAK1，JAK2和TYK2的Ki值分別為2.5，11，13和11nM。IC50&TargetJAK3JAK1Tyk2JAK22.5nM(Ki)11nM(Ki)11nM(Ki)13nM(Ki)FLT3ROCKIGSK3βCDK2/CycA1μM(Ki)1.5μM(Ki)1.8μM(Ki)2.6μM(Ki)PknB8μM(IC50)體外研究Decernotinib(VX-509)isapotentJAK3inhibitor,withKisof2.5,11,13and11nMforJAK3,JAK1,JAK2,andTYK2,respectively.DecernotinibpotentlyblocksT-cellproliferationwithameanIC50of170±101nM,andinhibitsIL-2-stimulatedT-cellproliferation(IC50,140and400nM).VX-509isalsocytotoxictoB-cellinresponsetoCD40LandIL-4(IC50,50nM)[1].體內(nèi)研究Decernotinib(VX-509,10,25,or50mg/kg,p.o.)significantlyanddose-dependentlyinhibitstheincreasesinanklediameterandpawweightoccuringinresponsetocollageninjectionsinrats.Decernotinibpotentlyalleviatescartilagedamageandboneresorptioninrats.Decernotinib(10,25,or50mg/kg,p.o.,b.i.d.)suppressesearedemainamousemodelofdelayed-typehypersensitivity[1].PROTOCOLKinaseAssay[1]TheeffectofDecernotinibonJAK3activityisassessedbymeasuringtheresidualkinaseactivityoftherecombinantlyexpressedJAK3kinasedomainusingaradiometricassay.Thefinalconcentrationsofthecomponentsintheassayareasfollows:100mMHEPES(pH7.5),10mMMgCl2,1mMdithiothreitol(DTT),0.01%BSA,0.25nMJAK3,0.25mg/mLpolyE4Y,and5μM33P-γ-ATP(200μCi/μmol).A10mMstocksolutionofDecernotinibispreparedinDMSO,fromwhichadditionaldilutionsareprepared.Asubstratemixture(100mMHEPES,10mMMgCl2,0.5mg/mLpolyE4Y,and10μM33P-γ-ATP)isaddedandmixedwithDecernotinibstocksolution.Thereactionisinitiatedbytheadditionofanenzymemixture[100mMHEPES(pH7.5),10mMMgCl2,2mMDTT,0.02%BSA,0.5nMJAK3].After15minutes,thereactionisquenchedwith20%trichloroaceticacid(TCA).ThequenchedreactionistransferredtotheGF/Bfilterplatesandwashedthreetimeswith5%TCA.FollowingtheadditionofUltimateGoldscintillant(50μL),thesamples2/4MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEarecountedinaPackardTopCountgammacounter.Inthisprocedure,theradioactivitytrappedisameasureoftheresidualJAK3kinaseactivity.FromtheactivityversusconcentrationofDecernotinibtitrationcurve,theKivalueisdeterminedbyfittingthedatatoanequationforcompetitivetightbindinginhibitionkinetics[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]Whole-bloodsamplesfromhealthyvolunteersareusedtocollectperipheralbloodmononuclearcells,whichareplatedinT75tissuecultureflasksatadensityof1×106/mL.Cellsarestimulatedwith10μg/mLphytohemagglutininat37°Cfor72hours.After72hours,cellsaredetachedfromtheflaskbyscraping,washed,andplatedatadensityof1×105/wellina96-wellplate.Decernotinib(9.7nMto10μM)isadded,andplatesareincubatedfor30minutesat37°C,followedbystimulationwithhumanIL-2.Intworows,onlyDMSOisadded;onerowisnotstimulatedwithIL-2,andonerowisstimulatedwithIL-2toserveastheproliferationcontrol.Platesareincubatedat37°Cfor2days.Onday2,cellsarepulsedwith20μCi/mLmethyl-[3H]thymidinefor18-24hoursandharvestedontofiltersforradiographicdetermination.DataareanalyzedtogenerateanIC50valueusingSoftmaxprosoftware[1]MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalRat[1]Administration[1]Thecollagen-inducedarthritis(CIA)ratmodelisusedtoevaluatetheeffectsoforalDecernotinib[10mg/kgb.i.d.,25mg/kgb.i.d.,50mg/kgb.i.d.,50mg/kgq.d.,or100mg/kgq.d.]onjointinflammationand

histopathology.FemaleLewisrats(157-187g)areanesthetizedwithisofluraneandinjectedwith300μL

Freund’sincompleteadjuvant,containing2mg/mLbovinetypeIIcollagen,atthebaseofthetailandtwo

sitesonthebackondays0and6.Theratsarerandomizedtostudygroupsattheonsetofpawswelling

(arthritis),whichoccursbetweendays10and11.DosingofeitherDecernotiniborvehicleviaoralgavageis

initiatedonthefirstdayofestablishedarthritisandcontinuedtoday6ofarthritis.Dosingvolumeis5mL/kg.

Groupsarecontrols(nocollageninjectionplusvehicle;n=4),collagenplusvehicle(n=5),collagenplus

Decernotinib10mg/kgb.i.d.(n=8);collagenplusDecernotinib10mg/kgb.i.d.(n=8);collagenplus(n=8);

collagenplus(n=8);collagenplus(n=8);collagenplu(n=8);andcollagenplus(n=8);collagenplu(n=

8);alltreatmentsareadministeredfor6days.Anadditionalgroupofratsisgivencollagenplus10mg/kg

subcutaneousetanercept,ahumantumornecrosisfactor-αantagonist,onstudydays11and14.Caliper

measurementsofnormal(baseline)anklejointsbeginonday9andcontinuethroughthelastdayofstudy.

DifferencesinmeananklediameteraretestedforsignificanceusingStudent’sttest,withsignificancesetat

P≤0.05.Theratsareeuthanizedonday7ofarthritis,whichisstudyday17or18dependingonwhen

animalsarerandomizedtogroups;pawsandkneesareharvestedtodeterminepawweightandtoconducta

histopathologicalanalysisofinflammation(kneeandankle),pannusformation(ankle),cartilagedestruction

(knee),andboneresorption(kneeandankle).Scoresrangefrom0(normal)to5(severepathology)andare

assignedbyaveterinarypathologist.Percentinhibitioniscalculatedusingthefollowingformula:[(meanof

treatmentgroup)?(meanofcontrol)]÷[(meanofcollagen+vehicle)?(meanofcontrol)].Kruskal-Wallis

one-wayanalysisofvariancenonparametrictestsareusedtodeterminestatisticalsignificanceamongthe

histopathologygroups,withsigni