Ruxolitinib-INCB18424-DataSheet-生命科學試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemERuxolitinibCat.No.:HY-50856CASNo.:941678-49-5Synonyms:INCB18424分?式:C??H??N?分?量:306.37作?靶點:JAK;Autophagy;Mitophagy;Apoptosis作?通路:Epigenetics;JAK/STATSignaling;ProteinTyrosineKinase/RTK;StemCell/Wnt;Autophagy;Apoptosis儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實驗DMSO:≥100mg/mL(326.40mM)H2O:<0.1mg/mL(ultrasonic;warming;heatto60°C)(insoluble)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM3.2640mL16.3201mL32.6403mL5mM0.6528mL3.2640mL6.5281mL10mM0.3264mL1.6320mL3.2640mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；?旦配成溶液，請分裝保存，避免反復凍融造成的產(chǎn)品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲存時，請在6個?內(nèi)使?，-20°C儲存時，請在1個?內(nèi)使?。體內(nèi)實驗請根據(jù)您的實驗動物和給藥?式選擇適當?shù)娜芙?案。以下溶解?案都請先按照InVitro?式配制澄的儲備液，再依次添加助溶劑：(為保證實驗結(jié)果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當保存；體內(nèi)實驗的?作液，建議您現(xiàn)?現(xiàn)1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE配，當天使?；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?；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的?式助溶)1.請依序添加每種溶劑：0.5%Methylcellulose/salinewaterSolubility:5mg/mL(16.32mM);Suspendedsolution;Needultrasonic2.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.08mg/mL(6.79mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(6.79mM);Clearsolution4.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(6.79mM);Clearsolution5.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(6.79mM);Clearsolution6.請依序添加每種溶劑：5%DMACin0.5%methylcelluloseaqueoussolutionSolubility:5mg/mL(16.32mM);Suspendedsolution;NeedultrasonicBIOLOGICALACTIVITY?物活性Ruxolitinib(INCB18424)有效，選擇性的JAK1/2抑制劑，IC50值分別為3.3nM和2.8nM，選擇性是JAK3的130多倍。Ruxolitinib誘導?噬(autophagy)，通過毒性線粒體?噬(mitophagy)殺死腫瘤細胞。IC50&TargetJAK2JAK1Tyk2JAK32.8nM(IC50)3.3nM(IC50)19nM(IC50)428nM(IC50)體外研究RuxolitinibpotentlyandselectivelyinhibitsJAK2V617F-mediatedsignalingandproliferation,markedlyincreasesapoptosisinadosedependentmanner,andat64nMresultsinadoublingofcellswithdepolarizedmitochondriainBa/F3cells.RuxolitinibdemonstratesremarkablepotencyagainsterythroidcolonyformationwithIC50of67nM,andinhibitsproliferatingoferythroidprogenitorsfromnormaldonorsandpolycythemiaverapatientswithIC50valuesof407nMand223nM,respectively[1].體內(nèi)研究Ruxolitinib(180mg/kg,orally,twiceaday)resultsinsurviverateofgreaterthan90%byday22andmarkedlyreducessplenomegalyandcirculatinglevelsofinflammatorycytokines,andpreferentiallyeliminatedneoplasticcells,resultinginsignificantlyprolongedsurvivalwithoutmyelosuppressiveorimmunosuppressiveeffectsinaJAK2V617F-drivenmousemodel[1].IntheRuxolitinibgroup,theprimaryendpointisreachedin41.9%ofpatients,ascomparedwith0.7%intheplacebogroupinthedouble-blindtrialofmyelofibrosis.Ruxolitinibresultsinmaintainingofreductioninspleenvolumeandimprovementof50%ormoreinthetotalsymptomscore[2].PROTOCOLKinaseAssay[1]RecombinantproteinsareexpressedusingSf21cellsandbaculovirusvectorsandpurifiedwithaffinitychromatography.JAKkinaseassaysuseahomogeneoustime-resolvedfluorescenceassaywiththepeptidesubstrate(-EQEDEPEGDYFEWLE).EachenzymereactioniscarriedoutwithRuxolitiniborcontrol,JAK2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEenzyme,500nMpeptide,adenosinetriphosphate(ATP;1mM),and2%dimethylsulfoxide(DMSO)for1hour.The50%inhibitoryconcentration(IC50)iscalculatedasINCB018424concentrationrequiredforinhibitionof50%ofthefluorescentsignal.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]Cellsareseededat2×103/wellofwhitebottom96-wellplates,treatedwithRuxolitinib(INCB018424)fromDMSOstocks(0.2%finalDMSOconcentration),andincubatedfor48hoursat37°Cwith5%CO2.ViabilityismeasuredbycellularATPdeterminationusingtheCell-TiterGloluciferasereagentorviablecellcounting.Valuesaretransformedtopercentinhibitionrelativetovehiclecontrol,andIC50curvesarefittedaccordingtononlinearregressionanalysisofthedatausingPRISMGraphPad.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMicearefedstandardrodentchowandprovidedwithwateradlibitum.Ba/F3-JAK2V617Fcells(105perAdministration[1]mouse)areinoculatedintravenouslyinto6-to8-week-oldfemaleBALB/cmice.Survivalismonitoreddaily,andmoribundmicearehumanelykilledandconsidereddeceasedattimeofdeath.Treatmentwithvehicle(5%dimethylacetamide,0.5%methocellulose)orRuxolitinib(INCB018424)beginwithin24hoursofcellinoculation,twicedailybyoralgavage.HematologicparametersaremeasuredusingaBayerAdvia120analyzed,andstatisticalsignificanceisdeterminedusingDunnetttesting.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻?NatMed.2018Aug;24(8):1143-1150.?Nature.2022Sep;609(7928):785-792.?Cell.2021Apr15;184(8):2167-2182.e22.?CancerDiscov.2018May;8(5):616-631.?Blood.2014Dec18;124(26):3924-31.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].Quintas-CardamaA,etal.PreclinicalcharacterizationoftheselectiveJAK1/2inhibi