SR9009-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•Agonists•ScreeningLibrarieswww.MedChemESR9009Cat.No.:HY-16989CASNo.:1379686-30-2分⼦式:C₂₀H₂₄ClN₃O₄S分⼦量:437.94作⽤靶点:Autophagy作⽤通路:Autophagy储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:≥100mg/mL(228.34mM)扫描⼆维码，H2O:<0.1mg/mL(insoluble)运⽤溶解⽅案计算器*"≥"meanssoluble,butsaturationunknown.获得适合您实验体系的溶解⽅案MassSolvent1mg5mg10mgConcentration制备储备液1mM2.2834mL11.4171mL22.8342mL5mM0.4567mL2.2834mL4.5668mL10mM0.2283mL1.1417mL2.2834mL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，并请注意储备液的保存⽅式和期限。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：1%DMSO99%saline2.Solubility:0.5mg/mL(1.14mM);Suspendedsolution;Needultrasonic请依序添加每种溶剂：10%DMSO40%PEG3005%Tween-8045%salineSolubility:2.08mg/mL(4.75mM);Suspendedsolution;Needultrasonic此⽅案可获得2.08mg/mL(4.75mM)的均匀悬浊液，悬浊液可⽤于⼝服和腹腔注射。1/3www.MedChemEwww.MedChemE以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到400μLPEG300中，混合均匀；向上述3.体系中加⼊50μLTween-80，混合均匀；然后继续加⼊450μL⽣理盐⽔定容⾄1mL。请依序添加每种溶剂：10%DMSO90%(20%SBE-β-CDinsaline)Solubility:2.08mg/mL(4.75mM);Suspendedsolution;Needultrasonic此⽅案可获得2.08mg/mL(4.75mM)的均匀悬浊液，悬浊液可⽤于⼝服和腹腔注射。以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到900μL20%的SBE-β-CD⽣理盐⽔⽔溶4.液中，混合均匀。请依序添加每种溶剂：10%DMSO90%cornoilSolubility:≥2.08mg/mL(4.75mM);Clearsolution此⽅案可获得≥2.08mg/mL(4.75mM，饱和度未知)的澄溶液，此⽅案不适⽤于实验周期在半个⽉以上的实验。5.以1mL⼯作液为例，取100μL20.8mg/mL的澄DMSO储备液加到900μL⽟⽶油中，混合均匀。请依序添加每种溶剂：5%DMSO40%PEG3005%Tween-8050%saline6.Solubility:2.5mg/mL(5.71mM);Suspendedsolution;Needultrasonic请依序添加每种溶剂：5%DMSO95%(20%SBE-β-CDinsaline)7.Solubility:2.5mg/mL(5.71mM);Suspendedsolution;Needultrasonic请依序添加每种溶剂：6%DMSO10%CremophorEL84%ddH2OSolubility:20mg/mL(45.67mM);Suspendedsolution;NeedultrasonicBIOLOGICALACTIVITY⽣物活性SR9009REV-ERBα/β激动剂，作⽤REV-ERBα和REV-ERBβ的IC50分别为670nM和800nM。IC50&TargetIC50:670nM(Rev-ErbBα),800nM(Rev-ErbBβ)[1]体外研究SR9009dose-dependentlyincreasestheREV-ERB-dependentrepressoractivityassessedinHEK293cellsexpressingachimericGal4DNABindingDomain(DBD)-REV-ERBligandbindingdomain(LBD)αorβandaGal4-responsiveluciferasereporter(SR9009:REV-ERBαIC50=670nM,REV-ERBβIC50=800nM).SR9009potentlyandefficaciouslysuppressestranscriptioninacotransfectionassayusingfull-lengthREV-ERBαalongwithaluciferasereporterdrivenbytheBmal1promoter(IC50=710nM).SR9009suppressestheexpressionofBMAL1mRNAinHepG2cellsinaREV-ERBα/β-dependentmanner.DirectbindingoftheSR9009toREV-ERBαisalsoconfirmedusingcirculardichrosimanalysis(Kd=800nM)[1].体内研究Whilethestressofhandlingandtwice-dailyinjectionscausedweightlossinvehicle-treatedcontrols,weightlossofSR9009-treatedanimalsis60%greater.SR9009(100mg/kg,i.p.)treatedmiceexhibitamoreseverereductioninadiposity.Plasmanon-esterifiedfattyacids(NEFA)arealsoreduced(23%)alongwithplasmaglucose(19%)intheSR9009treatedanimals.Inthewhiteadiposetissue(WAT),SR9009treatmentresultsinadecreaseinexpressionofgenesencodingenzymesinvolvedintriglyceride(TG)synthesisasisalsoobservedinleanmice[1].PROTOCOLCellAssay[1]HEK293cellsaregrownin96-wellplates(1×106/well)andaretransientlytransfectedusingLipofectamine.Cellsaretransfectedwithatotalof200ngofDNAperwellconsistingofthepGL4mIL-17fireflyluciferase2/3www.MedChemEwww.MedChemEreporterconstruct,thepGL4mIL-17+CNS-5fireflyluciferasereporterconstruct,orthepGL4mIL-172kBROREmutant(100ng/well),anactinpromoterRenillareniformisluciferasereporter(50ng/well),andeithercontrolvectoraloneorthetestDNA(full-lengthRORαorfull-lengthRORγat50ng/well).All48humannuclearreceptorsarerepresentedinthespecificityassayandSR9009istestedataconcentrationof20μM.TheformatoftheassayisacotransfectionassaywithGal4DNAbindingdomain-nuclearreceptorfusionsinHEK293cells[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1]ForcircadiangeneexpressionexperimentsmaleC57BL6mice(8-10weeksofage)areeithermaintainedonaL:D(12h:12h)cycleoronconstantdarkness.Atcircadiantime(CT)0animalsareadministeredasingledoseof100mg/kgSR9009orSR9011(i.p.)andgroupsofanimals(n=6)aresacrificedatCT0,CT6,CT12andCT18.GeneexpressionisdeterminedbyrealtimeQPCR.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•NatCommun.2018Oct12;9(1):4246.•JNeuroinflammation.2020Jan31;17(1):43.•Chemosphere.2021Jan;263:128020.•CellProlif.2021Jan13;e12988.•BiochemPharmacol.2020Feb;172:113773.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].SoltLA,etal.RegulationofcircadianbehaviourandmetabolismbysyntheticREV-ERBagonists.Nature.2012Mar29;485(7396):62-68.McePdfHeigh