GSK343-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEGSK343Cat.No.:HY-13500CASNo.:1346704-33-3分⼦式:C₃₁H₃₉N₇O₂分⼦量:541.69作⽤靶点:HistoneMethyltransferase;Autophagy作⽤通路:Epigenetics;Autophagy储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:15.62mg/mL(28.84mM;Needultrasonic)H2O:<0.1mg/mL(ultrasonic;warming;heatto60°C)(insoluble)MassSolvent1mg5mg10mgConcentration制备储备液1mM1.8461mL9.2304mL18.4607mL5mM0.3692mL1.8461mL3.6921mL10mM0.1846mL0.9230mL1.8461mL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.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%saline1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemESolubility:≥1.56mg/mL(2.88mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥1.56mg/mL(2.88mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥1.56mg/mL(2.88mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性GSK343⼀种⾼效,选择性的EZH2抑制剂，IC50为4nM。IC50&TargetIC50:4nM(EZH2),240nM(EZH1)[1]体外研究GSK343,whichcontainsann-propylgroupatthe4-positionofthepyridone,hasEZH2Kiapp=1.2±0.2nM.Inthis6-dayproliferationassay,amongthecelllinesevaluatedinthisstudy,theprostatecancercelllineLNCaPisthemostsensitivetoEZH2inhibition,withgrowthIC50valueof2.9μMforGSK343[1].GSK343isfoundtohavehalfmaximalinhibitoryconcentrationvaluesof13μMinHeLacellsand15μMinSiHacells[2].体内研究Comparewiththecontrols,GSK343(5mg/kg)-treatedmiceexhibitssignificantlyinhibitedtumorgrowth.TheaveragetumorvolumeandweightoftheGSK343-treatedcohortisremarkablyreduced.Asearlyas20dayspost-implantation,asignificantreductionintumorgrowthisobservedintheGSK343-treatedcohortrelativetothecontrolcohort;thisdifferencepersistedthroughtheremainderofthestudy.Inaddition,comparewiththecontrolcohort,theGSK343-treatedanimalsinthexenograftmodelshowaremarkableincreaseinmessengerRNAlevelsofE-cadherinbutasignificantdecreaseinvimentinmessengerRNAlevels[2].PROTOCOLKinaseAssay[1]ActivityagainstEZH2isassessedusing5memberPRC2complex(Flag-EZH2,EED,SUZ12,AEBP2,RbAp48).Theassayprotocolmaybesummarizedasfollows:10mMstocksofGSK343arepreparedfromsolidin100%DMSO.An11pointserialdilutionmasterplateispreparedin384wellformat(1:3dilution,columns6and18areequalvolumeDMSOcontrols)anddispensedtoassayreadyplatesusingacousticdispensingtechnologytocreatea100nLstampofGSK343andDMSOcontrols.Theassayadditionsconsistedofequalvolumeadditionsof10nMEZH2andthesubstratesolution(5μg/mLHeLanucleosomesand0.25μM[3H]-SAM)dispensedintoassayplatesusingamulti-dropcombidispense.Reactionplatesareincubatedfor1hrandquenchedwithanequalvolumeadditionof0.5mg/mLPS-PEIImagingBeads(RPNQ0098)containing0.1mMunlabeledSAM.Theplatesaresealed,darkadaptedfor30minutes,anda5minuteendpointluminescenceimageisacquiredusingaViewluximager.PlatestatisticssuchasZ’andsignaltobackgroundaswellasdoseresponsecurvesareanalyzedusingActivityBaseXE.TheinvitrobiochemicalactivityofEZH1isassessedaspartofa5memberPRC2complexusinga384wellSPAassayidenticaltoEZH2.Buffercomponents,reagentdispensing,GSK343platepreparation,quenchconditionsanddataanalysisareidenticalforEZH1andEZH2withfinalassayconcentrationsof20nMEZH1,5μg/mLHeLanucleosomesand0.25μM[3H]-SAM.Furtherdataanalysis,pIC50pivotsandvisualizationsareenabledbyTIBCOSpotfire[1].2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEMCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]Toaccountforvaryingdoublingratesamongcancercelllines,theoptimalcellseedingisdeterminedempiricallyforallcelllinesbyexaminingtheirgrowthina384-wellplateover6dayswithawiderangeofseedingdensities.Cellsarethenplatedattheoptimalseedingdensityandallowedtoadhereovernight.Cellsaretreatedinduplicatewitha20-point2-folddilutionseriesofGSK343or0.147%DMSO(vehiclecontrol)andincubatedfor6daysat37°Cin5%CO2.Cellsarethenlysedwith25μLCellTiter-GloperwellandchemiluminescenceisquantifiedwithaTECANSafire2microplatereader.Inaddition,anuntreatedplateofcellsisharvestedatthetimeofGSK343addition(T0)toquantifythestartingnumberofcells.CTGvaluesafter6daysoftreatmentareexpressedasapercentoftheT0valueandplottedagainstGSK343concentration.Dataarefitwitha4-parameterequationtogenerateaconcentrationresponsecurveandtheconcentrationofGSK343requiredtoinhibit50%ofgrowth(gIC50)isdetermined[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[2]Administration[2]Six-week-oldfemalenudeBALB/cmiceareused.TostudytheeffectoftheEZH2inhibitorGSK343,5mg/kgin100-μLphosphate-bufferedsalineisinjectedintraperitoneallyeveryotherdayintoBALB/cnudemice(n=6)afterthetumorvolumereaches100mm3.Inthisanalysis,thenegativecontrolgroup(n=6)receivedsaline.After40days,themicearekilled,andthesubcutaneoustumorsaresurgicallyexcised,weighed,photographed,sectioned,andfixedin10%formalin.TheexpressionlevelsofE-cadherin,N-cadherin,andvimentininthetumoursaremeasuredbyreal-timereversetranscriptionpolymerasechainreaction.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•CellStemCell.2019May2;24(5):769-784.e6.•JCachexiaSarcopeniaMuscle.2021Dec4.•Theranostics.2019Jul9;9(17):5020-5034.•Theranostics.201