ARS-853-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEARS-853Cat.No.:HY-19706CASNo.:1629268-00-3分⼦式:C₂₂H₂₉ClN₄O₃分⼦量:432.94作⽤靶点:Ras;Apoptosis作⽤通路:GPCR/GProtein;Apoptosis储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:25mg/mL(57.74mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.3098mL11.5489mL23.0979mL5mM0.4620mL2.3098mL4.6196mL10mM0.2310mL1.1549mL2.3098mL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；⼀旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存⽅式和期限：-80°C,6months;-20°C,1month。-80°C储存时，请在6个⽉内使⽤，-20°C储存时，请在1个⽉内使⽤。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：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(5.77mM);Clearsolution1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(5.77mM);Clearsolution3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(5.77mM);Clearsolution4.请依序添加每种溶剂：5%DMSO>>95%(20%SBE-β-CDinsaline)Solubility:≥2mg/mL(4.62mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性ARS-853选择性，共价的KRASG12C抑制剂，IC50为2.5μM。ARS-853通过与GDP结合的癌蛋⽩结合并阻⽌激活来抑制突变KRAS驱动的信号传导。IC50&TargetKRAS(G12C)2.5μM(IC50)体外研究ARS853isdesignedtobindKRASG12Cwithhighaffinity.TreatmentofKRASG12C-mutantlungcancercellswithARS853reducesthelevelofGTP-boundKRASbymorethan95%(10μM).ARS853inhibitsproliferationwithaninhibitoryconcentration50%(IC50)of2.5μM,whichissimilartoitsIC50fortargetinhibition.ARS853(10μM)inhibitseffectorsignalingandcellproliferationtovaryingdegreesinsixKRASG12Cmutantlungcancercelllines,butnotinnon-KRASG12Cmodels.Similarly,itcompletelysuppressestheeffectsofexogenousKRASG12CexpressiononKRAS-GTPlevels,KRAS-BRAFinteraction,andERKsignaling.ARS-853treatmentalsoinducesapoptosisinfourKRASG12Cmutantcelllines.ARS853selectivelyreducesKRAS-GTPlevelsandRAS-effectorsignalinginKRASG12C-mutantcells,whileinhibitingtheirproliferationandinducingcelldeath[1].ARS-853inhibitsmutantKRAS-drivensignalingbybindingtotheGDP-boundoncoproteinandpreventingactivation[2].PROTOCOLKinaseAssay[1]PurifiedKRAS(1μM)isincubatedEDTA(10mM)andGDP(1mM)orGTPγS(1mM)atroomtemperaturefor1hfollowedbyadditionofMgCl2(1mM)toterminatethereaction.ARS853(1μM)isthenaddedandthemixtureisincubatedforanotherhouratroomtemperature.HEK293cellsexpressingvariousKRASmutantsaretreatedwithARS853.Proteinsareextractedusingabuffercontaining9Murea,10mMDTTand50mMammoniumbicarbonate,pH8,heatedto65°Cfor15minandalkylatedusing50mMiodoacetamideat37°Cfor30min.ThesamplesaredesaltedbygelfiltrationinZebaspindesaltingplatesfollowedbyadditionofsequencing-gradetrypsintoaconcentrationof10μg/ml,andincubationforonehourat37°C.Heavyisotopicstandards(25fmol)oftheKRASG12CtargetpeptideandKRASnormalizationpeptideareaddedtothesamplesfollowedbydesaltinginStrata-Xpolymericreversephaseplates.LC-MS/MSanalysisisperformedinaQExactivequadrupoleorbitrapmassspectrometerunderstandardcondition.TheamountofKRASG12CboundbythedrugisdeterminedbytheratioofthemodifiedG12Cpeptidetothatoftheheavyisotopicstandards[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE户使⽤本产品发表的科研⽂献•CancerDiscov.2020Dec;10(12):1950-1967.•THEDEPARTMENTOFCANCERBIOLOGY.2020.28103931.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].LitoP,etal.Allele-specificinhibitorsinactivatemutantKRASG12Cbyatrappingmechanism.Science.2016Feb5;351(6273):604-8.[2].PatricelliMP,etal.SelectiveInhibitionofOncogenicKRASOutputwithSmallMoleculesTargetingtheInactiveState.CancerDiscov.2016Mar;6(3):316-29.M