SB225002-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•Agonists•ScreeningLibrarieswww.MedChemESB225002Cat.No.:HY-16711CASNo.:182498-32-4分⼦式:C₁₃H₁₀BrN₃O₄分⼦量:352.14作⽤靶点:CXCR作⽤通路:GPCR/GProtein;Immunology/Inflammation储存⽅式:4°C,storedundernitrogen*Insolvent:-80°C,6months;-20°C,1month(storedundernitrogen)溶解性数据体外实验DMSO:≥100mg/mL(283.98mM)扫描⼆维码，*"≥"meanssoluble,butsaturationunknown.运⽤溶解⽅案计算器获得适合您实验体系的溶解⽅案MassSolvent1mg5mg10mgConcentration制备储备液1mM2.8398mL14.1989mL28.3978mL5mM0.5680mL2.8398mL5.6796mL10mM0.2840mL1.4199mL2.8398mL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(7.10mM);Clearsolution此⽅案可获得≥2.5mg/mL(7.10mM，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(7.10mM);Clearsolution此⽅案可获得≥2.5mg/mL(7.10mM，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(7.10mM);Clearsolution此⽅案可获得≥2.5mg/mL(7.10mM，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SB225002⼀种有效的选择性CXCR2⾮肽拮抗剂，抑制125I-IL-8和CXCR2结合的IC50为22nM。IC50&Target125I-IL-8-CXCR222nM(IC50,inCHOcellmembrane)体外研究SB225002(SB225002)isanantagonistof125I-IL-8bindingtoCXCR2withanIC50=22nM.SB225002shows>150-foldselectivityoverCXCR1andfourother7-TMRstested.SB225002isapotentantagonistofrabbitCXCR2,inhibitingrabbitPMNchemotaxisinresponsetooptimalconcentrationsofhumanIL-8orGROα(IC50valuesof30and70nM,respectively.Inthesecells(PMN,HL60,CXCR1-RBL-2H3),SB225002producesaconcentration-dependentinhibitionofbothIL-8-andGROα-mediatedcalciummobilizationwithIC50valuesof8and10nM,respectively.In3ASubEcellsstablytransfectedwithCXCR2,SB225002dose-dependentlyinhibitscalciummobilizationinducedbybothGROαandIL-8,withIC50valuesof20and40nM,respectively[1].WHCO1cellstreatedwithSB225002exhibitsa40%reductionincellproliferation.BlockingCXCR2signalinginWHCO1cellswith400nMSB225002(SB225002)significantlydecreasescellproliferationby~40%to50%[2].体内研究SB225002(SB225002)selectivelyblocksIL-8-inducedneutrophilmarginationinrabbits[1].CXCR2isblockedusingtheselectiveantagonistSB225002(2mg/kg)orneutralizingCXCR2antiserum.TheCXCR2antagonistSB225002decreasesneutrophilcountsinischemichemispheresofApoE−/−miceonWesterndietandwildtypemiceonnormaldiet[3].SB225002significantlyattenuatesmicroglialactivationandBBBdamage,increasesmyelination,andreducesastrogliosisinthewhitematterafterLPS-sensitizedHI[4].PROTOCOLKinaseAssay[1]CHO-CXCR1andCHO-CXCR2membranesareprepared.Assaysareperformedin96-wellmicrotiterplateswherethereactionmixturecontained1.0μg/mLmembraneproteinin20mMBis-Tris-propane,pH8.0,with1.2mMMgSO4,0.1mMEDTA,25mMNaCl,and0.03%CHAPSandSB225002(10mMstockinMe2SO)addedattheindicatedconcentrations,thefinalMe2SOconcentrationis<1%understandardbindingconditions.Bindingisinitiatedbyadditionof0.25nM125I-IL-8(2,200Ci/mmol).After1-hincubationatroomtemperaturetheplateisharvestedusingaTomtec96-wellharvesterontoaglassfiberfiltermatblockedwith1%polyethyleneimine,0.5%BSAandwashedthreetimeswith25mMNaCl,10mMTris•HCl,1mMMgSO4,2/4www.MedChemEwww.MedChemE0.5mMEDTA,0.03%CHAPS,pH7.4.Thefilterisdried,sealedinasamplebagcontaining10mLofWallac205Betaplateliquidscintillationfluid,andcountedwithaWallac1205Betaplateliquidscintillationcounter[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[2]ThreeesophagealsquamouscellcarcinomacelllinesWHCO1,WHCO5,andWHCO6originallyestablishedfromsurgicalbiopsiesofprimaryesophagealsquamouscellcarcinomasareculturedinDMEMcontaining10%FCSat37°Cinahumidifiedatmosphereof5%CO2.MTTassaysarecarriedoutusingtheCellProliferationkit.Briefly,1.5×103cellsareplatedin96-wellplatesinafinalvolumeof180μLDMEMperwell.SB225002(400nM)isaddedtocellsand0.001%DMSO(solvent)isaddedasacontrol.Aftertheindicatedincubationperiod,18μLoftheMTTlabelingreagent(finalconcentration0.5mg/mL)isaddedtoeachwellandincubatedfor4hoursinahumidifiedatmosphere.Onehundredeightymicrolitersofthesolubilizationsolutionareaddedtoeachwellandtheplatesareleftovernightat37°C.Thespectrophotometricabsorbanceofsamplesismeasuredat595nmusingamicrotiterplatereader[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[3]Administration[3][4]Male7-8weeksoldwildtype(C57BL/6J,Harlan)andApoE−/−mice,whicharegeneratedonthesameC57BL/6background,areeitherfedwithanormalchoworacholesterolrichchowfor6weeksandsubmittedto20minofleft-sidedmiddlecerebralarteryocclusion(MCAO)orshamsurgery.Animalsarerandomlyattributedtotreatmentparadigms,andexperimentersareblindedatallstagesofinterventionsanddataanalysis.TheselectiveCXCR2antagonistSB225002(2mg/kg)orvehicle(1%DMSOinPBS)isinjectedintraperitoneally(i.p.)at0,24and48hourspost-ischemia.Inotherexperiments,CXCR2isspecificallyblockedbyi.p.injectionofaneutralizingrabbitanti-CXCR2serum(300μL)at0hours,24hoursand48hourspost-ischemia.Inthelatterstudies,normalrabbitserum(NRS)servedascontrol.Insomeexperiments,neutrophilsaredepletedbyi.p.injectionof200μganti-mouseLy6G24hoursbeforeand24hoursafterischemia.Intheseexperiments,200μgofanisotypecontrolantibodyisdeliveredascontrol.Rats[4]Inthisstudy,10-12Sprague-Dawleyratpupsperdamareused.Thepupsreceiveintraperitonealinjections

ofSB225002(1or3mg/kg,dilutedinNScontaining0.33%Tween80)orvehicle(NSsolutioncontaining0.33%Tween80)30minbeforelipopolysaccharide(LPS)administrationandimmediatelyafterhypoxic

ischemia(HI).Thepupsarerandomlyassignedtofourgroups:control(pupsunexposedtoLPSorHI,N=14),

vehicle(NSinjections30minbeforeLPSadministrationandimmediatelyafterHI,N=18),andSB-1(1mg/kg,

N=14)andSB-3(3mg/kg,N=18)(SB225002injections30minbeforeLPSadministrationandimmediately

afterHI).MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•NatCommun.2020Feb28;11(1):1126.•CancerRes.2018Oct1;78(19):5586-5599.•BioactMater.6(2021)2039-2057.•JAutoimmun.2018May;89:30-40.•JCellPhysiol.2021Apr;236(4):3114-3128.3/4www.MedChemEwww.MedChemESeemorecustomervalidationsonwww.MedChemEREFERENCES[1].WhiteJR,etal.Identificationofapotent,selectivenon-peptideCXCR2antagonistthatinhibitsinterleukin-8-inducedneutrophilmigration.JBiolChem.1998Apr24;273(17):10095-8.[2].WangB,etal.Agrowth-relatedoncogene/CXCchemokinereceptor2autocrineloopcontributestocellularproliferationinesophagealcancer.CancerRes.20