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(diǎn):JAK;Autophagy;Mitophagy;Apoptosis作?通路:Epigenetics;JAK/STATSignaling;ProteinTyrosineKinase/RTK;StemCell/Wnt;Autophagy;Apoptosis儲(chǔ)存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:≥100mg/mL(326.40mM)H2O:<0.1mg/mL(ultrasonic;warming;heatto60°C)(insoluble)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM3.2640mL16.3201mL32.6403mL5mM0.6528mL3.2640mL6.5281mL10mM0.3264mL1.6320mL3.2640mL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。-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)1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE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.請(qǐng)依序添加每種溶劑：0.5%Methylcellulose/salinewaterSolubility:5mg/mL(16.32mM);Suspendedsolution;Needultrasonic2.請(qǐng)依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.08mg/mL(6.79mM);Clearsolution3.請(qǐng)依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.08mg/mL(6.79mM);Clearsolution4.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(6.79mM);Clearsolution5.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(6.79mM);Clearsolution6.請(qǐng)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(dǎo)?噬(autophagy)，通過(guò)毒性線粒體?噬(mitophagy)殺死腫瘤細(xì)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(xiàn)?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