Ruxolitinib-INCB18424-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemERuxolitinibCat.No.:HY-50856CASNo.:941678-49-5Synonyms:INCB18424分⼦式:C₁₇H₁₈N₆分⼦量:306.37作⽤靶点:JAK;Autophagy;Mitophagy;Apoptosis作⽤通路:Epigenetics;JAK/STATSignaling;ProteinTyrosineKinase/RTK;StemCell/Wnt;Autophagy;Apoptosis储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:≥100mg/mL(326.40mM)H2O:<0.1mg/mL(ultrasonic;warming;heatto60°C)(insoluble)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制备储备液1mM3.2640mL16.3201mL32.6403mL5mM0.6528mL3.2640mL6.5281mL10mM0.3264mL1.6320mL3.2640mL请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；⼀旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存⽅式和期限：-80°C,6months;-20°C,1month。-80°C储存时，请在6个⽉内使⽤，-20°C储存时，请在1个⽉内使⽤。体内实验请根据您的实验动物和给药⽅式选择适当的溶解⽅案。以下溶解⽅案都请先按照InVitro⽅式配制澄的储备液，再依次添加助溶剂：(为保证实验结果的可靠性，澄的储备液可以根据储存条件，适当保存；体内实验的⼯作液，建议您现⽤现1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE配，当天使⽤；以下溶剂前显⽰的百分⽐指该溶剂在您配制终溶液中的体积占⽐；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的⽅式助溶)1.请依序添加每种溶剂：0.5%Methylcellulose/salinewaterSolubility:5mg/mL(16.32mM);Suspendedsolution;Needultrasonic2.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.08mg/mL(6.79mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(6.79mM);Clearsolution4.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(6.79mM);Clearsolution5.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(6.79mM);Clearsolution6.请依序添加每种溶剂：5%DMACin0.5%methylcelluloseaqueoussolutionSolubility:5mg/mL(16.32mM);Suspendedsolution;NeedultrasonicBIOLOGICALACTIVITY⽣物活性Ruxolitinib(INCB18424)有效，选择性的JAK1/2抑制剂，IC50值分别为3.3nM和2.8nM，选择性是JAK3的130多倍。Ruxolitinib诱导⾃噬(autophagy)，通过毒性线粒体⾃噬(mitophagy)杀死肿瘤细胞。IC50&TargetJAK2JAK1Tyk2JAK32.8nM(IC50)3.3nM(IC50)19nM(IC50)428nM(IC50)体外研究RuxolitinibpotentlyandselectivelyinhibitsJAK2V617F-mediatedsignalingandproliferation,markedlyincreasesapoptosisinadosedependentmanner,andat64nMresultsinadoublingofcellswithdepolarizedmitochondriainBa/F3cells.RuxolitinibdemonstratesremarkablepotencyagainsterythroidcolonyformationwithIC50of67nM,andinhibitsproliferatingoferythroidprogenitorsfromnormaldonorsandpolycythemiaverapatientswithIC50valuesof407nMand223nM,respectively[1].体内研究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—您⾝边的抑制剂⼤师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.户使⽤本产品发表的科研⽂献•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