Salubrinal-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•Agonists•ScreeningLibrarieswww.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(protectfromlight)溶解性数据体外实验DMSO:≥50mg/mL(104.21mM)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1mM2.0842mL10.4208mL20.8416mL5mM0.4168mL2.0842mL4.1683mL10mM0.2084mL1.0421mL2.0842mL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，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：10%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.5mg/mL(5.21mM);Clearsolution此⽅案可获得≥2.5mg/mL(5.21mM，饱和度未知)的澄溶液。以1mL⼯作液为例，取100μL25.0mg/mL的澄DMSO储备液加到400μLPEG300中，混合均匀；向上述2.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)1/4www.MedChemEwww.MedChemESolubility:2.5mg/mL(5.21mM);Suspendedsolution;Needultrasonic此⽅案可获得2.5mg/mL(5.21mM)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100μL25.0mg/mL的澄DMSO储备液加到900μL20%的SBE-β-CD⽣理盐⽔⽔溶3.液中，混合均匀。请依序添加每种溶剂：10%DMSO90%cornoilSolubility:≥2.5mg/mL(5.21mM);Clearsolution此⽅案可获得≥2.5mg/mL(5.21mM，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。以1mL⼯作液为例，取100μL25.0mg/mL的澄DMSO储备液加到900μL⽟⽶油中，混合均匀。BIOLOGICALACTIVITY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&TargetDusp2HSV-1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%2/4www.MedChemEwww.MedChemETween20),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.AnimalMice[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.•CancerBiolMed.2019Feb;16(1):38-54.•CancerBiolMed.2019Feb;16(1):38-54.•FrontCellDevBiol.2020May12;8:269.•BiomedPharmacother.2020May;125:109819.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].DrexlerHC.SynergisticApoptosisInductioninLeukemicCellsbythePhosphataseInhibitorSalubrinalandProteasomeInhibitors.PLoSOne.2009;4(1