GSK126-GSK2816126A-DataSheet-生命科學試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEGSK126Cat.No.:HY-13470CASNo.:1346574-57-9Synonyms:GSK2816126A分?式:C??H??N?O?分?量:526.67作?靶點:HistoneMethyltransferase作?通路:Epigenetics儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數據體外實驗DMSO:12.5mg/mL(23.73mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲備液1mM1.8987mL9.4936mL18.9872mL5mM0.3797mL1.8987mL3.7974mL10mM0.1899mL0.9494mL1.8987mL請根據產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，避免反復凍融造成的產品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲存時，請在6個?內使?，-20°C儲存時，請在1個?內使?。體內實驗請根據您的實驗動物和給藥?式選擇適當的溶解?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：(為保證實驗結果的可靠性，澄的儲備液可以根據儲存條件，適當保存；體內實驗的?作液，建議您現(xiàn)?現(xiàn)配，當天使?；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.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%saline1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemESolubility:≥1.25mg/mL(2.37mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:1.25mg/mL(2.37mM);Suspendedsolution;Needultrasonic3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥1.25mg/mL(2.37mM);Clearsolution4.請依序添加每種溶劑：20%SBE-β-CDadjustedtopH4-4.5with1NaceticSolubility:20mg/mL(37.97mM);Clearsolution;Needultrasonicandwarming5.請依序添加每種溶劑：5%DMSO>>40%PEG300>>5%Tween-80>>50%salineSolubility:≥0.82mg/mL(1.56mM);Clearsolution6.請依序添加每種溶劑：5%DMSO>>95%(20%SBE-β-CDinsaline)Solubility:≥0.82mg/mL(1.56mM);Clearsolution7.請依序添加每種溶劑：50%PEG300>>50%salineSolubility:10mg/mL(18.99mM);Suspendedsolution;NeedultrasonicBIOLOGICALACTIVITY?物活性GSK126(GSK2816126A)?種有效，選擇性的EZH2甲轉移酶抑制劑，IC50為9.9nM。IC50&TargetEZH29.9nM(IC50)體外研究GSK126potentlyinhibitsbothwild-typeandmutantEZH2methyltransferaseactivitywithsimilarpotencies(Ki=0.5-3nM)independentofsubstrateused,andiscompetitivewithS-adenosyl-methionine(SAM)andnon-competitivewithpeptidesubstrates.GSK126ishighlyselectiveagainstothermethyltransferasesandmultipleotherproteinclasses(EZH1,IC50=680nM)[1].TreatmentofthreeSCLCcelllineswithGSK126,inducesgrowthinhibition.SCLCcelllines(Lu130,H209,andDMS53)aretreatedwith0.5,2,and8μMGSK126,andgrowthcurveisanalyzedbyWST-8assay.InhibitionofcellulargrowthbyGSK126treatmentisobservedat8μMinallthethreecelllines,whileLu130andH209aremoresensitivetoGSK126,evenatlowerdoses[2].體內研究GSK126isadministeredintraperitoneallyatadosevolumeof0.2mLper20gbodyweightinfemalebeigeSCIDmice.GSK126effectivelyinhibitstheproliferationofEZH2mutantDLBCLcelllinesandmarkedlyinhibitsthegrowthofEZH2mutantDLBCLxenograftsinmice[1].PROTOCOLKinaseAssay[1]Thefive-memberPRC2complex(Flag-EZH2,EED,SUZ12,AEBP2,RbAp48)containingeitherwild-typeormutant(A677G,Y641N,Y641C,Y641H,Y641SorY641F)EZH2isprepared.GSK126isdissolvedinDMSOandtestedatconcentrationsof0.6nMto300nMwithafinalDMSOconcentrationof2.5%.Incontrasttowild-typeEZH2whichprefersH3K27me0asasubstrateinvitro,EZH2Y641mutantspreferH3K27me2andhavelittleactivitywithH3K27me0orH3K27me1.TheA677Gmutantisdistinctfromboththewild-typeandY641mutantformsofEZH2inthatitefficientlymethylatesH3K27me0,H3K27me1,andH3K27me2;2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEtherefore,histoneH3peptides(residues21-44;10μMfinal)witheitherK27me0(wildtype,A677GEZH2),K27me1(A677GEZH2),orK27me2(A677G,Y641N,Y641C,Y641H,Y641SandY641FEZH2)areusedasmethyltransferasesubstrates.GSK126isaddedtoplatesfollowedbyadditionof6nMEZH2complexandpeptide.AsthepotencyofGSK126isatornearthetightbindinglimitofanassayrunat[SAM]=Km,IC50valuesaremeasuredatahighconcentrationofthecompetitivesubstrateSAMrelativetoitsKm(7.5μMSAMwheretheSAMKmis0.3μM)[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[2]JUB-andPTRF-introducedDMS53cellsareseededatdensityof1×103cells/wellin96-wellplate,andcellulargrowthisanalyzedusingWST-8kitat12,36,60,and84h.CellulargrowthofLu130,H209,andDMS53withtreatmentbyDZNeporGSK126isalsoanalyzedusingWST-8kit.DZNepisdissolvedinPBSat5mM,andcellsareculturedatthefinalconcentrationof5μM.GSK126isdissolvedinDMSOat10mM,andcellsareculturedat0.5,2,and8μM[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1]GSK126orvehicleisadministeredintraperitoneallyatadosevolumeof0.2mLper20gbodyweight.PfeifferorKARPAS-422cells(1×107)in100%MatrigelareimplantedsubcutaneouslyinfemalebeigeSCIDmice.Tumorsaremeasuredwithcalipers,andblockrandomizedaccordingtotumoursizeintotreatmentgroups.Forefficacystudies,10micearerandomizedineachtreatmentgroupbeforetheinitiationofdosingandGSK126treatmentisinitiatedoncethetumourvolumesareapproximately200mm3inthePfeifferandKARPAS-422studiesand500mm3intheKARPAS-422intermittentdosingstudy.Miceareweighedandtumorsmeasuredwithcaliperstwiceweekly.Two-tailedt-testsareconductedassumingtwosamplesofequalvariance.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產品發(fā)表的科研?獻?Cell.2018Sep20;175(1):186-199.e19.?Blood.2022Aug19;blood.2022015668.?JExpMed.2018May7;215(5):1365-1382.?JExpClinCancerRes.2021Oct19;40(1):330.?CellDeathDiffer.2022May14.Seemore