Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEBETd-260Cat.No.:HY-101519CASNo.:2093388-62-4Synonyms:ZBC260分?式:C??H??N??O?分?量:798.89作?靶點:PROTACs;EpigeneticReaderDomain;Apoptosis作?通路:PROTAC;Epigenetics;Apoptosis儲存?式:-80°C,protectfromlight,storedundernitrogen溶解性數(shù)據(jù)體外實驗DMSO:25mg/mL(31.29mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲備液1mM1.2517mL6.2587mL12.5174mL5mM0.2503mL1.2517mL2.5035mL10mM0.1252mL0.6259mL1.2517mL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(protectfromlight,storedundernitrogen)。-80°C儲存時，請在6個?內(nèi)使?。體內(nèi)實驗請根據(jù)您的實驗動物和給藥?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：(為保證實驗結(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)實驗的?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.請依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥0.83mg/mL(1.04mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥0.83mg/mL(1.04mM);Clearsolution1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEBIOLOGICALACTIVITY?物活性BETd-260(ZBC260)由Cereblon配體和BET配體相連的PROTAC，在??病細胞RS4;11中，在30pM的低濃度下，能夠降低BRD4蛋?活性。BETd-260能顯著抑制肝細胞癌HCC細胞的活性并誘導(dǎo)細胞凋亡(apoptosis)。IC50&TargetBRD4BRD2BRD350)30-100pM(IC50)30-100pM(IC50)體外研究BETd-260(ZBC260;Compound23)iscapableofinducingdegradationofBRD2,BRD3,andBRD4proteinsat30–100pMintheRS4;11leukemiacells.BETd-260showsinhibitoryactivityagainstthegrowthofRS4;11leukemiacellsandMOLM-13cellswithIC50sof51pMand2.2nM,respectively,andinducesapoptosisinbothRS4;11andMOLM-13celllinesat3-10nM[1].BETd-260reciprocallymodulatestheexpressionofseveralapoptoticgenesinHCCcells,i.e.,suppressingtheexpressionofanti-apoptoticMcl-1,Bcl-2,c-Myc,andXIAP,whereasincreasingtheexpressionofpro-apoptoticBad[2].體內(nèi)研究BETd-260(5mg/kg,i.v.,everyotherday,thriceaweekfor3weeks)causesrapidtumorregressionwithamaximumof>90%regressioninmicebearingRS4;11xenografttumors,andwithnobodyweightlossorothersignsoftoxicityinmice.BETd-260(5mg/kg,i.v.)degradestheBRD2,BRD3,andBRD4proteinsformorethan24h,withrobustcleavageofPARPandcaspase-3,andstrongdown-regulationofc-MycproteininRS4;11xenograftmicemodel[1].PROTOCOLCellAssay[1]Incellgrowthexperiments,cellsareseededin96-wellcellcultureplatesatadensityof10000?20000cells/wellin100μLofculturemedium.BETd-260isseriallydilutedintheappropriatemedium,and100μLofthedilutedsolutioncontainingBETd-260isaddedtotheappropriatewellsofthecellplate.AfteradditionofBETd-260,thecellsareincubatedfor4daysat37°Cinanatmosphereof5%CO2.Cellgrowthisevaluatedbyalactatedehydrogenase-basedWST-8assayusingamultimodemicroplatereader.TheWST-8reagentisaddedtotheplate,incubatedforatleast1h,andreadat450nm.ThereadingsarenormalizedtotheDMSO-treatedcells,andtheIC50iscalculatedbynonlinearregressionanalysisusingGraphPadPrism6software[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1]Todevelopxenografttumors,5×106RS4;11cellswith50%Matrigelareinjectedsubcutaneouslyonthedorsalsideofseverecombinedimmunodeficient(SCID)mice,onetumorpermouse.Whentumorsreachappr100mm3,micearerandomlyassignedtoBETd-260treatmentandvehiclecontrolgroups.Animalsaremonitoreddailyforanysignsoftoxicityandweighed2-3timesperweekduringthetreatmentandweighedatleastweeklyafterBETd-260treatmentend.Tumorsizeismeasured2-3timesperweekbyelectroniccalipersduringthetreatmentperiodandatleastweeklyafterthetreatmentisend.Tumorvolumeis2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEcalculatedasV=LW2/2,whereListhelengthandWisthewidthofthetumor[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].ZhouB,etal.DiscoveryofaSmall-MoleculeDegraderofBromodomainandExtra-Terminal(BET)ProteinswithPicomolarCellularPotenciesandCapableofAchievingTumorRegression.JMedChem.2018Jan25;61(2):462-481.[2].ZhangH,etal.TargetingBETProteinsWithaPROTACMoleculeElicitsPotentAnticancerActivityinHCCCells.FrontOncol.2020;9:1471.Publish