Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEARS-853Cat.No.:HY-19706CASNo.:1629268-00-3分?式:C??H??ClN?O?分?量:432.94作?靶點(diǎn):Ras;Apoptosis作?通路:GPCR/GProtein;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:25mg/mL(57.74mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制備儲(chǔ)備液1mM2.3098mL11.5489mL23.0979mL5mM0.4620mL2.3098mL4.6196mL10mM0.2310mL1.1549mL2.3098mL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?。體內(nèi)實(shí)驗(yàn)請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥?式選擇適當(dāng)?shù)娜芙?案。以下溶解?案都請(qǐng)先按照InVitro?式配制澄的儲(chǔ)備液，再依次添加助溶劑：(為保證實(shí)驗(yàn)結(jié)果的可靠性，澄的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)保存；體內(nèi)實(shí)驗(yàn)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(qǐng)依序添加每種溶劑：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.5mg/mL(5.77mM);Clearsolution1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE2.請(qǐng)依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:≥2.5mg/mL(5.77mM);Clearsolution3.請(qǐng)依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(5.77mM);Clearsolution4.請(qǐng)依序添加每種溶劑：5%DMSO>>95%(20%SBE-β-CDinsaline)Solubility:≥2mg/mL(4.62mM);ClearsolutionBIOLOGICALACTIVITY?物活性ARS-853選擇性，共價(jià)的KRASG12C抑制劑，IC50為2.5μM。ARS-853通過與GDP結(jié)合的癌蛋?結(jié)合并阻?激活來抑制突變KRAS驅(qū)動(dòng)的信號(hào)傳導(dǎo)。IC50&TargetKRAS(G12C)2.5μM(IC50)體外研究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?邊的抑制劑?師www.MedChemE戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?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