Doramapimod-BIRB-796-DataSheet-生命科学试剂-MedChemExpress

Hotline:400-820-3792Inhibitors•ScreeningLibraries•Proteinswww.MedChemEDoramapimodCat.No.:HY-10320CASNo.:285983-48-4Synonyms:BIRB796分⼦式:C₃₁H₃₇N₅O₃分⼦量:527.66作⽤靶点:p38MAPK;Raf;Autophagy作⽤通路:MAPK/ERKPathway;Autophagy储存⽅式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性数据体外实验DMSO:125mg/mL(236.89mM;Needultrasonic)Ethanol:33.33mg/mL(63.17mM;Needultrasonic)MassSolvent1mg5mg10mgConcentration制备储备液1mM1.8952mL9.4758mL18.9516mL5mM0.3790mL1.8952mL3.7903mL10mM0.1895mL0.9476mL1.8952mL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。-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)1/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemE1.请依序添加每种溶剂：10%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.08mg/mL(3.94mM);Clearsolution2.请依序添加每种溶剂：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:2.08mg/mL(3.94mM);Suspendedsolution;Needultrasonic3.请依序添加每种溶剂：10%DMSO>>90%cornoilSolubility:≥2.5mg/mL(4.74mM);ClearsolutionBIOLOGICALACTIVITY⽣物活性Doramapimod(BIRB796)⼀种具有⼝服活性的，⾼效的p38MAPK抑制剂，IC50值分别为p38α=38nM，p38β=65nM，p38γ=200nM，p38δ=520nM。Doramapimod对p38激酶具有⽪尔亲和⼒(Kd=0.1nM)。Doramapimod也抑制B-Raf和Abl，IC50分别为83nM和14.6μM[1][2]。IC50&Targetp38αp38βp38δp38γ38nM(IC50)65nM(IC50)520nM(IC50)200nM(IC50)B-RafAblp38MAPkinase83.4nM(IC50)14600nM(IC50)0.1nM(Kd)体外研究Doramapimod(BIRB796)isusuallyassociatedwithinflammationbecauseofitsroleinT-cellproliferationandcytokineproduction[1].Doramapimod(BIRB796)blocksthestress-inducedphosphorylationofthescaffoldproteinSAP97,furtherestablishingthatthisisaphysiologicalsubstrateofSAPK3/p38γ.ThebindingofDoramapimodtothep38MAPKsorJNK1/2isimpairingtheirphosphorylationbytheupstreamkinaseMKK6orMKK4[3].体内研究ThemeanxenograftweighofDoramapimod(BIRB796)islighterthancontrol.TheinhibitionrateofDoramapimodis1.93%[4].TheDoramapimod(BIRB796)treatmentslightlyreducesbloodpressure(166±7mmHgatweek7;P[5].PROTOCOLCellAssay[3]Humanembryonickidney(HEK)293andHeLacellsareexposedto0.5Msorbitolfor30minor100ng/mLEGFfor10minandthenlysedinbufferA(50mMTris-HCl,pH7.5,1mMEGTA,1mMEDTA,1mMsodiumorthovanadate,10mMsodiumfluoride,50mMsodiumβ-glycerophosphate,5mMpyrophosphate,0.27Msucrose,0.1mMphenylmethylsulfonylfluoride,1%(v/v)TritonX-100)plus0.1%(v/v)2-mercaptoethanolandCompleteproteinaseinhibitormixture.Lysatesarecentrifugedat18,000×gfor5minat4°C,andthesupernatantsareremoved,quick-frozeninliquidnitrogen,andstoredat-20°Cuntiluse.Whenrequired,cellsarepreincubatedfor1hwithoutorwith10μMSB203580or10μMPD184352orwithdifferentconcentrationsofDoramapimodforthetimesindicatedinthefigures[3].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[4]2/3MasterofBioactiveMolecules—您⾝边的抑制剂⼤师www.MedChemEAdministration[4][5]Athymicnudemice(BALB/c-nu/nu),6to8weeksofageandweighing18to24g,areused.ThemicearetreatedwithDoramapimod(10mg/kgp.o.,every3days×5).Thebodyweightsoftheanimalsandthetwoperpendiculartumordiameters(AandB)arerecordedevery3days,andthetumorvolume(V)isestimated.Rats[5]MaletransgenicdTGRs(RCCLtd)andage-matchednontransgenicSprague-Dawley(SD)rats(MDC)areuse.2differentprotocolsareperformed.Inprotocol2,untreateddTGR(n=15),dTGR+BIRB796(30mg/kgperdayinthedietfor3weeks;n=11),andSD(n=8eachgroup)ratsareanalyzed.Systolicbloodpressureismeasuredweeklybytailcuff.Twenty-four-hoururinesamplesarecollectedinmetaboliccagesfromweeks5to7.Serumiscollectedatweek7.SerumcreatinineandcystatinCaremeasuredbyclinicalroutineassays.Urinaryratalbuminisdeterminedbyenzyme-linkedimmunosorbentassay.Theaimofprotocol2istofocusonelectrophysiologicalalterationsandmortality.UntreateddTGR(n=10),dTGR+BIRB796(n=10),andSD(n=10)ratsarestudieduptoweek8.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.户使⽤本产品发表的科研⽂献•Nature.2019Jul;571(7763):127-131.•CellRes.2020Jul;30(7):574-589.•SciTranslMed.2018Jul18;10(450).pii:eaaq1093.•DevCell.2021Dec20;56(24):3334-3348.e6.•ExpMolMed.2021Sep21.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].DietrichJ,etal.Thedesign,synthesis,andevaluationof8hybridDFG-outallosterickinaseinhibitors.BioorgMedChem.2010Aug1;18(15):5738-48[2].CicenasJ,etal.JNK,p38,ERK,andSGK1InhibitorsinCancer.Cancers(Basel).2017Dec21;10(1).pii:E1.[3].KumaY,etal.BIRB796inhibitsallp38MAPKisoformsinvitroandinvivo.JBiolChem,2005,280(20),19472-19479.[4].HeD,etal.BIRB796,theinhibitorofp38mitogen-activatedproteinkinase,enhancestheefficacyofchemotherapeuticagentsinABCB1overexpressioncells.PLoSOne.2013;8(1):e54181.[5].ParkJK,etal.p38mitogen-activatedproteinkinaseinhibitionamelioratesangiotensinI