Litronesib-LY2523355-DataSheet-生命科學(xué)試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemELitronesibCat.No.:HY-14846CASNo.:910634-41-2Synonyms:LY2523355分?式:C??H??N?O?S?分?量:511.7作?靶點(diǎn):Kinesin作?通路:CellCycle/DNADamage;Cytoskeleton儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:≥50mg/mL(97.71mM)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM1.9543mL9.7713mL19.5427mL5mM0.3909mL1.9543mL3.9085mL10mM0.1954mL0.9771mL1.9543mL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(fù)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?jiān)?個?內(nèi)使?，-20°C儲存時，請?jiān)?個?內(nèi)使?。體內(nèi)實(shí)驗(yàn)請根據(jù)您的實(shí)驗(yàn)動物和給藥?式選擇適當(dāng)?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(shí)驗(yàn)結(jié)果的可靠性，澄的儲備液可以根據(jù)儲存條件，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)1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE1.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(4.89mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(4.89mM);Clearsolution3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(4.89mM);ClearsolutionBIOLOGICALACTIVITY?物活性Litronesib(LY2523355)?種選擇性的有絲分裂特異性運(yùn)動蛋?Eg5抑制劑，具有抗腫瘤的活性[1]。IC50&TargetEg5體外研究Litronesib(LY2523355)isaselectiveEg5inhibitor.Litronesib(25nM)inducescancercellstodeathduringmitoticarrest,andthisneedssustainedactivationofspindle-assemblycheckpoint(SAC)[1].體內(nèi)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?邊的抑制劑?師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