Salubrinal-DataSheet-生命科學試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?Agonists?ScreeningLibrarieswww.MedChemESalubrinalCat.No.:HY-15486CASNo.:405060-95-9分?式:C??H??Cl?N?OS分?量:479.81作?靶點:Phosphatase;HSV;Autophagy;Apoptosis作?通路:MetabolicEnzyme/Protease;Anti-infection;Autophagy;Apoptosis儲存?式:4°C,protectfromlight*Insolvent:-80°C,6months;-20°C,1month(protectfromlight)溶解性數(shù)據(jù)體外實驗DMSO:≥50mg/mL(104.21mM)掃描?維碼，H2O:<0.1mg/mL(insoluble)運?溶解?案計算器*"≥"meanssoluble,butsaturationunknown.獲得適合您實驗體系的溶解?案MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.0842mL10.4208mL20.8416mL5mM0.4168mL2.0842mL4.1683mL10mM0.2084mL1.0421mL2.0842mL請根據(jù)產品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存?式和期限。體內實驗請根據(jù)您的實驗動物和給藥?式選擇適當?shù)娜芙?案。以下溶解?案都請先按照InVitro?式配制澄的儲備液，再依次添加助溶劑：為保證實驗結果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當保存；體內實驗的?作液，建議您現(xiàn)?現(xiàn)配，當天使?；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.請依序添加每種溶劑：10%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.5mg/mL(5.21mM);Clearsolution此?案可獲得≥2.5mg/mL(5.21mM，飽和度未知)的澄溶液。以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲備液加到400μLPEG300中，混合均勻；向上述2.體系中加?50μLTween-80，混合均勻；然后繼續(xù)加?450μL?理鹽?定容?1mL。請依序添加每種溶劑：10%DMSO90%(20%SBE-β-CDinsaline)1/4www.MedChemEwww.MedChemESolubility:2.5mg/mL(5.21mM);Suspendedsolution;Needultrasonic此?案可獲得2.5mg/mL(5.21mM)的均勻懸濁液，懸濁液可?于?服和腹腔注射。以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲備液加到900μL20%的SBE-β-CD?理鹽??溶3.液中，混合均勻。請依序添加每種溶劑：10%DMSO90%cornoilSolubility:≥2.5mg/mL(5.21mM);Clearsolution此?案可獲得≥2.5mg/mL(5.21mM，飽和度未知)的澄溶液，此?案不適?于實驗周期在半個?以上的實驗。以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲備液加到900μL??油中，混合均勻。BIOLOGICALACTIVITY?物活性Salubrinal有效的選擇性eIF2α去磷酸化抑制劑。Salubrinal作為雙特異性磷酸酶2(Dusp2)抑制劑，抑制抗膠原蛋?抗體誘導的關節(jié)炎[2]。Salubrinal具有抗HSV-1病毒的活性，并抑制由HSV-1蛋?ICP34.5介導的eIF2α的去磷酸化。IC50&TargetDusp2HSV-1體外研究Salubrinal,arecentlyidentifiedPP1inhibitorcapabletoprotectagainstendoplasmicreticulum(ER)stressinvariousmodelsystems,stronglysynergizedwithproteasomeinhibitorstoaugmentapoptoticdeathofdifferentleukemiccelllines.SalubrinalpreferentiallyseemstotargetthePP1/GADD34complex,SalubrinalisofinteresttoexaminewhethertheeffectofSalubrinalcouldalsoberecapitulatedbyanotherinhibitorofthisphosphatase.Forthispurposecantharidin,wisselected,whichislesstoxicthanokadaicacid,butwhichalsoblocksPP1(IC50=1.7μM)activities[1].體內研究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.戶使?本產品發(fā)表的科研?獻?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