ML346-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•Agonists•ScreeningLibrarieswww.MedChemEML346Cat.No.:HY-18669CASNo.:100872-83-1分⼦式:C₁₄H₁₂N₂O₄分⼦量:272.26作⽤靶点:HSP作⽤通路:CellCycle/DNADamage;MetabolicEnzyme/Protease储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:12.5mg/mL(45.91mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM3.6730mL18.3648mL36.7296mL5mM0.7346mL3.6730mL7.3459mL10mM0.3673mL1.8365mL3.6730mL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，配制前请先配制澄的储备液，再依次添加助溶剂(为保证实验结果的可靠性，体内实验的⼯作液，建议您现⽤现配，当天使⽤；澄的储备液可以根据储存条件，适当保存；以下溶剂前的百分⽐指该溶剂在您配制终溶液中的体积占⽐)：1.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥1.25mg/mL(4.59mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性ML346Hsp70和HSF-1活性的激活剂，针对Hsp70的EC50为4.6μM。ML346恢复构象疾病模型中的蛋1/3MasterofSmallMolecules—您⾝边的抑制剂⼤师www.MedChemE⽩质折叠，⽽没有明显的细胞毒性或缺乏特异性。ML346诱导了热休克反应(HSR)的因和蛋⽩质效应⼦的特异性增加，包括伴侣蛋⽩如Hsp70，Hsp40和Hsp27。IC50&TargetHSP704.6μM(EC50,HeLacells)体外研究ML346isanactivatorofHsp70,withanEC50of4600nMinHeLacells.ML346(10μM)restoresproteostasis,restoresCFTR-mediatediodideconductance,andenhancesthecorrectfoldingofproteinsexpressedintwodifferentcellularcompartments[1].ML346(CompoundF1)inducesmultipleresponsesandstronglyinducesHsp70,theoxidativestressresponsegenes(HO1andGCLM),anda2.5-foldupregulationofBiPinWTMEFcells.ML346(0.5-25μM)exhibitscytoprotectiveeffectsincellsaftera35minsevereheatshock,andalsocausesatwo-foldprotectionfromH2O2-inducedapoptosis[2].体内研究ML346suppresstheaggregationofpolyQ35inaC.elegansmodel,suggestingtheprobehasefficacyinmodifyingproteinaggregationandassociatedtoxicity[1].PROTOCOLKinaseAssay[2]Inbrief,HeLacellsareincubatedwitheitherDMSO(negativecontrol),thepositivecontrolsMG132(10μM)andlactacystin(6μM)orthePRsA1,A3andML346(F1)for3and6hoursandthenharvested.Cellsarelysedinhomogenizationbuffer(50mMTris-HCl,pH7.5,250mMsucrose,5mMMgCl2,2mMATP,1mMDTT,0.5mMEDTA,0.025%digitonin)for5minonice,andtotalproteinconcentrationofwholecellextractisdetermined.3μgofwholecellextractsarecombinedwithassaybuffer(50mMTris-HCl,pH7.5,40mMKCl,5mMMgCl2,0.5mMATP,1mMDTT,0.05mg/mLBSA)inablack96-wellplateandthereactionisinitiatedbytheadditionofa2×(200μM)fluorogenicpeptidesubstrateSuc-LLVY-AMC.Fluorescenceismeasuredevery10minusingaSynergyH4multi-modemicroplatereader[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[2]HeLacellsareplatedatadensityof10,000cellsperwellinblack96-wellplatesin100μLofDMEMsupplementedwith10%FBSand1%Pen/Strep/Neo.Platesareincubatedfor16hoursat37°C,5%CO2and95%relativehumiditybeforecompoundaddition.1μLofhitcompounds(ML346)inDMSOorDMSOaloneareaddedtothesampleorcontrolwells,respectively.Platesarethenplacedbackintheincubatorfor24hours.Afterincubation,cellsarewashed2×with200μLofPBSand200μLofasolutionof1μg/mLofcalceinAMisaddedtoeachwell.Cellsarethenincubatedfor45minat37°C,5%CO2beforefluorescencemeasurementusinganAnalystGTmultimodereader.PercentcytotoxicityisexpressedrelativetowellscontainingcellstreatedwithDMSOonly(100%)[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.REFERENCES[1].CalaminiB,etal.ML346:ANovelModulatorofProteostasisforProteinConformationalDiseases.ProbeReportsfromtheNIHMolecularLibrariesProgram.Bethesda(MD):NationalCenterforBiotechnologyInformation(US);2010-.2012Dec17.[2].CalaminiB,etal.Small-moleculeproteostasisregulatorsforproteinconformationaldiseases.NatChemBiol.2011Dec25;8(2):185-96.2/3MasterofSmallMolecules—您⾝边的抑制剂⼤师www.MedChemEMcePd