SP600125-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•Agonists•ScreeningLibrarieswww.MedChemESP600125Cat.No.:HY-12041CASNo.:129-56-6分⼦式:C₁₄H₈N₂O分⼦量:220.23作⽤靶点:JNK;Autophagy;Apoptosis;Ferroptosis作⽤通路:MAPK/ERKPathway;Autophagy;Apoptosis储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:12.5mg/mL(56.76mM;Needultrasonic)扫描⼆维码，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1mM4.5407mL22.7035mL45.4071mL5mM0.9081mL4.5407mL9.0814mL10mM0.4541mL2.2704mL4.5407mL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(11.35mM);Clearsolution此⽅案可获得≥2.5mg/mL(11.35mM，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1/4www.MedChemEwww.MedChemE2.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)Solubility:≥0.2mg/mL(0.91mM);Clearsolution此⽅案可获得≥0.2mg/mL(0.91mM，饱和度未知)的澄溶液。以1mL⼯作液为例，取100μL2.0mg/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:≥0.2mg/mL(0.91mM);Clearsolution此⽅案可获得≥0.2mg/mL(0.91mM，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100μL2.0mg/mL的澄DMSO储备液加到900μL⽟⽶油中，混合均匀。BIOLOGICALACTIVITY⽣物活性SP600125⼀种⼝服有效的，可逆的，ATP竞争性的JNK抑制剂，抑制JNK1，JNK2和JNK3的IC50分别为40，40，90nM。SP600125⼀种有效的铁死亡(ferroptosis)抑制剂。SP600125抑制⾃噬(autophagy)，诱导凋亡(apoptosis)。IC50&TargetJNK1JNK2JNK3Autophagy40nM(IC50)40nM(IC50)90nM(IC50)体外研究SP600125isanATP-competitiveinhibitorofJNK2withaKivalueof0.19±0.06μM.SP600125inhibitsthephosphorylationofc-JunwithIC50of5μMto10μMinJurkatTcells.InCD4+cells,suchasTh0cellsisolatedfromeitherhumancordorperipheralblood,SP600125blockscellactivationanddifferentiationandinhibitstheexpressionofinflammatorygenesCOX-2,IL-2,IL-10,IFN-γ,andTNF-α,withIC50of5μMto12μM[1].InamousebetacellsMIN6,SP600125(20μM)inducesthephosphorylationofp38MAPKanditsdownstreamCREB-dependentpromoteractivation[2].InHCT116cells,SP600125(20μM)blockstheG2phasetomitosistransitionandinducesendoreplication.ThisabilityofSP600125isindependentofJNKinhibition,butduetoitsinhibitionofCDK1-cyclinBactivationupstreamofAuroraAandPolo-likekinase1[3].体内研究AdministrationofSP600125at15or30mg/kgi.v.significantlyinhibitsTNF-αserumlevels,whereasoraladministrationdose-dependentlyblocksTNF-αexpressionwithsignificantinhibitionobservedat30mg/kgperos[1].SP600125attenuatesLPS-inducedALIinratsinvivo.TheexpressionlevelsofTNF-αandIL-6intheBALFinratsintheSP600125grouparesignificantlydecreased[4].PROTOCOLCellAssay[1]DeterminationofmRNAhalf-lifeisperformedessentially,exceptthatCD14+cellsarestimulatedwith(bacterial)lipopolysaccharide(LPS;50ng/mL)for2hbeforeadditionofactinomycinD(5μg/mL).SP600125(25μM)orvehicle(0.5%DMSOvol/vol)isaddedimmediatelyfollowingtheactinomycinD.Analysisis2/4www.MedChemEwww.MedChemEperformedbyusingreal-timereversetranscription(RT)-PCR.TotalRNAisextractedwithanRNeasyMinikit.TNFmRNAismeasuredbyrealtimeRT-PCR,usingaTNFTaqmanprobe.Alldataarenormalizedbyusingglyceraldehyde-3-phosphatedehydrogenase(GAPDH)expression.TheTNF-αforwardprimeris5′-CTGGCCCAGGCAGTCAGAT-3′andthereverseprimeris5′-TATCTCTCAGCTCCACGCCATT-3′.TheTaqmanprobesequenceis5′-FAM-CCTGTAGCCCATGTTGTAGCAAACCCTCA-TAMRA-3′[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[1]Administration[1][4]FemaleCD-1mice(8-10weeksofage)aredosedi.v.orperoswithSP600125inPPCESvehicle(30%PEG-400/20%polypropyleneglycol/15%CremophorEL/5%ethanol/30%saline),finalvolumeof5mL/kg,15minbeforei.v.injectionwithLPSinsaline(0.5mg/kg).At90min,aterminalbleedisobtainedfromtheabdominalvenacava,andtheserumisrecovered.SamplesareanalyzedformouseTNF-αbyusinganELISA.Rats[4]Atotalof40maleWistarratsarerandomlydividedintofourgroups(n=10):thecontrolgroup,LPSgroup,normalsalinegroup(NS)andtheSP600125group.Acutelunginjury(ALI)isinducedviaintratrachealinjectionofLPS.NormalsalineorSP600125isadministeredviaintraperitonealinjection(15mg/kg)10minaftertheLPSinjection.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•SciTranslMed.2019Feb6;11(478).pii:eaau5266.•SciTranslMed.2018Jul18;10(450).pii:eaaq1093.•MolCell.2020Jan2;77(1):95-107.e5.•NatCommun.2020Jan3;11(1):71.•CellDeathDiffer.2020Jun;27(6):1938-1951.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].BennettBL,etal.SP600125,ananthrapyrazoloneinhibitorofJunN-terminalkinase.ProcNatlAcadSciUSA,2001,98(24),13681-13686.[2].VaishnavD,etal.SP600125,aninhibitorofc-junN-terminalkinase,activatesCREBbyap38MAPK-mediatedpathway.BiochemBiophysResCommun,2003,307(4),855-860.[3].KimJA,etal.SP600125suppressesCdk1andinducesendoreplicationdirectlyfromG2phase,independentofJNKinhibition.Oncogene,2010,29(11),1702-1716.[4].ZhengY,etal.JNKinhibitorSP600125protectsagainstlipopolysaccharide-inducedacutelunginjuryviaupregulationofclaudin-4.ExpTherMed.2014Jul;8(1):153-158.[5].ZhangH,e