Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEROC-325Cat.No.:HY-103706CASNo.:1859141-26-6分?式:C??H??ClN?OS分?量:503.06作?靶點(diǎn):Autophagy;Apoptosis作?通路:Autophagy;Apoptosis儲(chǔ)存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMSO:32mg/mL(63.61mM;Needultrasonic)H2O:1mg/mL(1.99mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM1.9878mL9.9392mL19.8783mL5mM0.3976mL1.9878mL3.9757mL10mM0.1988mL0.9939mL1.9878mL請(qǐng)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(chǔ)備液；?旦配成溶液，請(qǐng)分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲(chǔ)備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?，-20°C儲(chǔ)存時(shí)，請(qǐng)?jiān)?個(gè)?內(nèi)使?。BIOLOGICALACTIVITY?物活性ROC-325?種有效的具有?服活性的?噬(autophagy)抑制劑，具有很強(qiáng)的抗癌活性。ROC-325引起溶酶體脫酸，?噬體積累和?噬通量中斷。ROC-325還誘導(dǎo)腎細(xì)胞癌凋亡(apoptosis)。IC50&TargetAutophagy[1]1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE體外研究ROC-325antagonizesrenalcellcarcinoma(RCC)growthandsurvivalinanATG5/7-dependentmanner,inducesapoptosis,andexhibitsfavorableselectivity.ROC-325inhibitscellsgrowthwithIC50valuesof4.9μM,11μM,4.6μM,5.4μM,7.4μM,11μM,8.2μM,5.8μM,5.0μM,11μM,8.4μMand6.0μMforA498,A549,CFPAC-1,COLO-205,DLD-1,IGROV-1,MCF-7,MiaPaCa-2,NCI-H69,PC-3,RLandUACC-62cells,respectively.ROC-325induceshallmarkfeaturesofautophagyinhibitionandantagonizesautophagicflux[1].ROC-325triggersahighlysignificantincreaseincathepsinD(CTSD)levels.Treatmentwith5μMROC-325for24hoursleadstotheformationofLC3BpunctaeandarobustincreaseinLC3BlevelsinbothA498and786-0RCCcells.ImmunoblottinganalysisconductedinbothA498and786-0cellsdemonstratesthatROC-325promotesadose-dependentincreaseinLC3Bexpressioninamannerthatcorrelatedwithacorrespondingincreaseinthelevelsofp62andcathepsinD[1].體內(nèi)研究OraladministrationofROC-325(25mg/kg,40mg/kg,50mg/kg,)tomicebearing786-0RCCxenograftsiswelltolerated,significantlymoreeffectiveatinhibitingtumorprogressionthanHydroxychloroquine,andinhibitsautophagyinvivo[1].PROTOCOLKinaseAssay[1]RenalcancercellsareincubatedwithROC-325for24hours.Cellsareharvestedandthenlysed.Approximately50μgoftotalcellularproteinfromeachsamplearesubjectedtoSDS,proteinsaretransferredtonitrocellulosemembranes,andthemembranesareblockedwith5%nonfatmilkinaTris-bufferedsalinesolutioncontaining0.1%Tween-20for1hour.Theblotsarethenprobedovernightat4°Cwithprimaryantibodies,washed,andprobedwithspecies-specificsecondaryantibodiescoupledtohorseradishperoxidase.Immunoreactivematerialisdetectedbyenhancedchemiluminescence[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]CellviabilityisestimatedbytheMTTassay.Cellsareseededinto96-wellmicrocultureplatesat10,000cellsperwellandallowedtoattachfor24hours.CellsarethentreatedwithROC-325for72hours.FollowingROC-325treatment,MTTisaddedandformazanabsorbanceisquantifiedusingamicroplatereader.Theestimatedcellviabilityundereachexperimentalconditioniscalculatedbynormalizingtherespectiveformazanopticaldensitytothedensityofcontrolcells.ProapoptoticeffectsfollowinginvitroROC-325exposurearequantifiedbypropidiumiodide(PI)stainingandfluorescence-activatedcellsorting(FACS)analysisofsub-G0/G1DNAcontentandbymeasurementofactivecaspase-3byflowcytometryusingacommercialkit[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.Animal786-0renalcancercells(5×106)aresuspendedinamixtureofHBSSandMatrigelandsubcutaneouslyAdministration[1]implantedintofemalenudemice.Tumor-bearinganimalsfromeachcelllinexenograftarerandomizedintotreatmentgroups.Micearetreatedwithvehicle(water),ROC-325(25,40,and50mg/kgPO)QD×5for6weeks.Micearemonitoreddailyandtumorvolumesaremeasuredtwiceweekly.Atstudycompletion,tumorsfromrepresentativeanimalsareexcisedfromeachgroup,formalin-fixed,andparaffin-embeddedforimmunohistochemicalanalysis[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE[1].CarewJS,etal.DisruptionofAutophagicDegradationwithROC-325AntagonizesRenalCellCarcinomaPathogenesis.ClinCancerRes.2017Jun1;