Salubrinal-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemESalubrinalCat.No.:HY-15486CASNo.:405060-95-9分⼦式:C₂₁H₁₇Cl₃N₄OS分⼦量:479.81作⽤靶点:Phosphatase;HSV;Autophagy;Apoptosis作⽤通路:MetabolicEnzyme/Protease;Anti-infection;Autophagy;Apoptosis储存⽅式:4°C,protectfromlight*Insolvent:-80°C,6months;-20°C,1month(protectfrom

light)溶解性数据体外实验DMSO:50mg/mL(104.21mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM2.0842mL10.4208mL20.8416mL5mM0.4168mL2.0842mL4.1683mL10mM0.2084mL1.0421mL2.0842mL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(protectfromlight)。-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：45%PEG300>>5%Tween-80>>50%salineSolubility:10mg/mL(20.84mM);Suspendedsolution;Needultrasonic1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEBIOLOGICALACTIVITY⽣物活性Salubrinal有效的选择性eIF2α去磷酸化抑制剂。Salubrinal作为双特异性磷酸酶2(Dusp2)抑制剂，抑制抗胶原蛋⽩抗体诱导的关节炎[2]。Salubrinal具有抗HSV-1病毒的活性，并抑制由HSV-1蛋⽩ICP34.5介导的eIF2α的去磷酸化。IC50&TargetHSV-1Dusp2体外研究Salubrinal,arecentlyidentifiedPP1inhibitorcapabletoprotectagainstendoplasmicreticulum(ER)stressinvariousmodelsystems,stronglysynergizedwithproteasomeinhibitorstoaugmentapoptoticdeathofdifferentleukemiccelllines.SalubrinalpreferentiallyseemstotargetthePP1/GADD34complex,SalubrinalisofinteresttoexaminewhethertheeffectofSalubrinalcouldalsoberecapitulatedbyanotherinhibitorofthisphosphatase.Forthispurposecantharidin,wisselected,whichislesstoxicthanokadaicacid,butwhichalsoblocksPP1(IC50=1.7µM)activities[1].体内研究Salubrinalisasyntheticchemicalthatinhibitsde-phosphorylationofeukaryotictranslationinitiationfactor2alpha(eIF2α).SalubrinalsignificantlysuppressesinflammationofthepawsofCAIAmice.Forinstance,theclinicalscoresare1.94±1.7(placebo)and0.31±0.6(Salubrinal)onday6;and4.63±3.4(placebo)and1.09±1.6(Salubrinal)onday12.Consistentwiththeclinicalscores,thethickeningofthepawsisalsoreducedintheSalubrinal-treatedgroup.Furthermore,Salubrinalreducesthehistologicalscoresfrom1.47±1.10(N=16;placebo)to0.59±0.64(N=16;Salubrinal)(p=0.01)[2].PROTOCOLKinaseAssay[1]Phosphataseactivitiesaredeterminedonimmunoprecipitatesofthephosphatases.Briefly,2×106K562cellsaretreatedfor18hrwithSalubrinal(20µM),PSI(10nM),thecombinationofbothdrugsorokadaicacid(100nM).AfterwashingwithPBS,cellsarelysedfor15minoniceeitherinPP1LB(fordeterminationofPP1γ-activity;20mMTris-HCl,pH7.5,1%TritonX-100,10%glycerol,132mMNaCl,Rochecompleteproteaseinhibitor)orinRIPA(forPP2A),supplementedwithRochecompleteproteaseinhibitor).Celllysatescontaining500µg(PP1γ)or300µg(PP2A)proteinareimmunoprecipitatedovernightat4°Cwith2-3µgoftheappropriateantibodiesandthenincubatedwithProteinA-Sepharose.Immunoprecipitatesarewashedthreetimesinlysisbuffer,followedbyresuspensioninphosphataseassaybuffer(PP2A:20mMTris-HCl,pH7.5,0.1mMCaCl2;PP1γ:50mMTrisHClpH7.0,0.2mMMnCl2,0.1mMCaCl2,125µg/mLBSA,0.05%Tween20),supplementedwith100µM6,8-difluoro-4-methyl-umbelliferylphosphate(DiFMUP).Precipitatesareallowedtoreactwithsubstratefor1hrat37°ConanEppendorfThermoshaker,centrifugedandDiFMUfluorescenceismeasuredonaBioTekLambdaFluoro320microplatereader(360nmex/460nmem).Phosphataseactivitiesaregivenaspercentchangerelativetothecontrol(DMSOtreatedcells)[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]CellularviabilityisassessedbytheWST-1colorimetricassay.Assaysareperformedon96wellplateswith2×104K562cells/wellintriplicatewithSalubrinalconcentrationsrangingfrom5-75µM(totalvolumeof200µL,18hrs).Untreatedcellsservedasnegativecontrolsample[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEAnimalMice[2]Administration[2]UsingBalb/cfemalemice(~nineweeksold),CAIAisinducedbyintravenousinjectionofa2mgcocktailofArthritoMAbantibodiesonday0followedbyintraperitonealinjectionof100µgLPSonday3.MicearerandomlydividedintoaplacebogroupandaSalubrinal-treatedgroup.Salubrinal(2.0mg/kg)isintravenouslyadministereddailyfromday0,whileasolvent(49.5%PEG400and0.5%Tween80inPBS)isadministeredtotheplacebogroup.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•ActaBiomater.2020Jun;109:229-243.•EMBORep.2022Apr11;e53932.•CellProlif.2021Sep28;e13133.•BiomedPharmacother.2020May;125:109819.•EcotoxicolEnvironSaf.2022Aug15;243:113966.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].DrexlerHC.SynergisticApoptosisInductioninLeukemicCellsbythePhosphataseInhibitorSalubrinalandProteasomeInhibitors.PLoSOne.2009;4(1):e4161.[2].HamamuraK,etal.SalubrinalactsasaDusp2inhibitorandsuppressesinflammationinanti-collagenantibody-inducedarthritis.CellSignal.2015Apr;27(4):828-35.[3].BryantKF,etal.ICP34.5-dependentand-independentactivitiesofsalubrinalin