Litronesib-LY2523355-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemELitronesibCat.No.:HY-14846CASNo.:910634-41-2Synonyms:LY2523355分⼦式:C₂₃H₃₇N₅O₄S₂分⼦量:511.7作⽤靶点:Kinesin作⽤通路:CellCycle/DNADamage;Cytoskeleton储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:≥50mg/mL(97.71mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制备储备液1mM1.9543mL9.7713mL19.5427mL5mM0.3909mL1.9543mL3.9085mL10mM0.1954mL0.9771mL1.9543mL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)1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE1.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(4.89mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(4.89mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(4.89mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性Litronesib(LY2523355)⼀种选择性的有丝分裂特异性运动蛋⽩Eg5抑制剂，具有抗肿瘤的活性[1]。IC50&TargetEg5体外研究Litronesib(LY2523355)isaselectiveEg5inhibitor.Litronesib(25nM)inducescancercellstodeathduringmitoticarrest,andthisneedssustainedactivationofspindle-assemblycheckpoint(SAC)[1].体内研究Litronesib(LY2523355;1.1,3.3,10,and30mg/kg,i.v.)showsantitumoractivityinadose-dependently,andcausesadramaticincreaseincancercellsimmuno-positiveforhistoneH3phosphorylationinColo205xenografttumors[1].PROTOCOLCellAssay[1]Cancercellsareplatedinpoly-d-lysinecoated96-wellplatesandincubatedovernight.CellsarethentreatedwithindicatedconcentrationsofLitronesibforvarioustimeperiods.Cellsarethenfixedwith3.7%formaldehydeinPBSfor45minutesor1×Preferfixativesolutionfor30minutes,atroomtemperature,andpermeabilizedwithcoldmethanolfor10minutesandthen0.2%TritonX-100inPBSfor10minutes.CellsarewashedthreetimeswithPBS.Cellsarethenincubatedwith100μg/mLDNase-freeRNaseand10μg/mLofpropidiumiodidefor1hourandscannedformitoticindex(MI)measurementbasedonDNAcondensationaspercentageofcellswithcondensedDNA.ForMIbasedonhistoneH3phosphorylationorapoptosisanalysis,cellsareincubatedovernightat4°Cwithanti-phospho-histoneH3antibodyoranti-phospho-histoneH2AXat1:1,000dilutionwith5%bovineserumalbumin(BSA)inPBS,respectively.Cellsarewashedthreetimeswith0.2%Triton-X100inPBSandincubatedwithAlexa488secondaryantibody(1:1,000inPBS-2%BSA)for60minutesatroomtemperature.CellsarewashedthreetimesagainwithPBSandstainedfor15minutesinPBScontaining10μg/mLpropidiumiodideand100μg/mLRNaseA.ThestainedcellsarescannedwithanAcumenExplorereX3microplatecytometer.Theresultsareexpressedaspercentageofcellspositiveforphospho-histoneH3-Ser10orphospho-histoneH2AX[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalPrimaryhuman-tumorxenograftmodelsareestablishedandmaintainedinnudemice.AntitumorefficacyinAdministration[1]subcutaneousxenografttumor-bearingmicewith10micepertreatmentgroupfromeitherestablishedcancercelllinesorfragmentsofhumantumorexplantsisevaluatedastumorvolumebyserialcalipermeasurementsandiscalculated.Thep388syngeneictumormodelisdevelopedforhigh-contentimaging2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEanalysisusingfemaleBDF1mice,weighing20to23g,whichareacclimatedin-houseforoneweekbeforetheiruseinexperiments.p388murinelymphocyticleukemiacellsareauthenticatedbySTRassayandmaintainedinRPMI1640mediumcontaining10%FBS.Forinoculation,thecellsarewashedwithserum-freemediumthreetimesand1.25millioncellsareimplantedbyintraperitonealinjectionintomice.Onday5afterimplantation,micearetreatedwithLitronesib,viaeitherintravenousbolusorintravenousinfusionatappropriatedosesanddurations.Miceareeuthanizedandtheascitic(intraperitoneal)fluidcontainingthep388tumorcellsisdrawnandanalyzedbyacumen,flowcytometry,andTUNELassaysforphospho-histoneH3,G2-M,andapoptosis.Forpharmacokineticstudy,thebloodsamplesarecollectedviacardiacpunctureandgeneratedplasmawithEDTAanddeterminedLitronesibexposureintheplasma[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].YeXS,etal.ANovelEg5Inhibitor(LY2523355)CausesMitoticArrestandApoptosisinCancerCellsandShowsPotentAntitumorActivityinXenograftTumorModels.M