Hotline:400-820-3792Inhibitors?ScreeningLibraries?Proteinswww.MedChemEAGI-5198Cat.No.:HY-18082CASNo.:1355326-35-0Synonyms:IDH-C35分?式:C??H??FN?O?分?量:462.56作?靶點(diǎn):IsocitrateDehydrogenase(IDH)作?通路:MetabolicEnzyme/Protease儲存?式:Powder-20°C3years4°C2yearsInsolvent-80°C6months-20°C1month溶解性數(shù)據(jù)體外實(shí)驗(yàn)DMF:≥50mg/mL(108.09mM)DMSO:20.83mg/mL(45.03mM;Needultrasonic)*"≥"meanssoluble,butsaturationunknown.MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.1619mL10.8094mL21.6188mL5mM0.4324mL2.1619mL4.3238mL10mM0.2162mL1.0809mL2.1619mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；?旦配成溶液，請分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲備液的保存?式和期限：-80°C,6months;-20°C,1month。-80°C儲存時(shí)，請?jiān)?個(gè)?內(nèi)使?，-20°C儲存時(shí)，請?jiān)?個(gè)?內(nèi)使?。體內(nèi)實(shí)驗(yàn)請根據(jù)您的實(shí)驗(yàn)動物和給藥?式選擇適當(dāng)?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(shí)驗(yàn)結(jié)果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當(dāng)保存；體內(nèi)實(shí)驗(yàn)的?作液，建議您現(xiàn)?現(xiàn)配，當(dāng)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1/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemE、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%DMSO>>40%PEG300>>5%Tween-80>>45%salineSolubility:≥2.08mg/mL(4.50mM);Clearsolution2.請依序添加每種溶劑：10%DMSO>>90%(20%SBE-β-CDinsaline)Solubility:2.08mg/mL(4.50mM);Suspendedsolution;Needultrasonic3.請依序添加每種溶劑：10%DMSO>>90%cornoilSolubility:≥2.08mg/mL(4.50mM);ClearsolutionBIOLOGICALACTIVITY?物活性AGI-5198(IDH-C35)有效和選擇性的突變體IDH1R132H抑制劑，IC50為0.07μM。IC50&TargetIDH1體外研究MeasurementsofR-2HGconcentrationsinpelletsofTS603gliomacellsdemonstratesdose-dependentinhibitionofthemutantIDH1enzymebyAGI-5198.AGI-5198doesnotimpaircolonyformationoftwopatient-derivedgliomalinesthatexpressonlythewild-typeIDH1allele(TS676andTS516)[1].CancercellsheterozygousfortheIDH1(R132H)mutationexhibitslessIDH-mediatedproductionofNADPH,suchthatafterexposuretoionizingradiation(IR),therearehigherlevelsofreactiveoxygenspecies,DNAdouble-strandbreaks,andcelldeathcomparedwithIDH1wild-typecells.TheseeffectsarereversedbytheIDH1(R132H)inhibitorAGI-5198[2].體內(nèi)研究AGI-5198(450mg/kg,p.o.)causes50to60%growthinhibitionofthetumorgrowthfromhumangliomaxenografts.TumorsfromAGI-5198-treatedmiceshowreducedstainingwithanantibodyagainsttheKi-67protein.AGI-5198doesnotaffectthegrowthofIDH1wild-typegliomaxenografts[1].PROTOCOLKinaseAssay[1]InhibitorypotencyagainsttheIDH2R140QandIDH2R172KenzymesisdeterminedinanendpointassayinwhichtheamountofNADPHremainingattheendofthereactionismeasuredbytheadditionofalargeexcessofdiaphoraseandresazurin.IDH2R140Qisdilutedto0.25μg/mLin40μL1XAssayBuffer(150mMNaCl,50mMpotassiumphosphatepH7.5,10mMMgCl2,10%glycerol,2mMB-ME,0.03%BSA)andincubatedfor16hoursat25°Cinthepresenceof1μLofcompoundinDMSO.Thereactionisstartedwiththeadditionof10μLofSubstrateMix(20μMNADPH,8μMalpha-ketoglutarate,in1XAssayBuffer)andincubatedfor1hourat25°C.Then,remainingNADPHismeasuredbytheadditionof25μLofDetectionMix(36μg/mLdiaphorase,18μMresazurinin1XAssaybuffer),incubatedfor5minutesat25°C,andreadasdescribedabove.IDH2R172KisassayedasforIDH2R140Qwiththefollowingmodifications:1.25μg/mLofproteinisused,theSubstrateMixcontained50μMNADPHand6.4μMalpha-ketoglutarate,andthecompoundisincubatedfor1hourbeforestartingthereaction.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[1]TS603cellsaregrowninmediumcontainingeitherAGI-5198(1.5μM)orDMSOvehiclecontrol.Oneweek2/3MasterofBioactiveMolecules—您?邊的抑制劑?師www.MedChemEpriortoharvestcellsareransferredtodifferentiationmedium(DMEMF12;15mMHEPES;0.06%glucose;B27withoutvitaminA;N2;Insulin/transferrin;1%FBS)containingfreshlyaddedretinoicacid(1μM).ChIPofnon-crosslinkedcellsisthencarriedoutusingestablishedChIPmethods.350μgoflysateisimmunoprecipitated-usinganti-H3K9Me3,H3K27me3orRabbitControlIgG.Afterwashing,ChIPDNAiselutedfromproteinGbeadsandanalyzedbyRT-PCRusingSYBRgreen.RelativeoccupancyiscalculatedusingthestandardcurvemethodandfoldenrichmentversusIgG.EnrichmentinAGI-5198-treatedcellsisnormalizedtovehiclecontrol.Meansandstandarddeviationarecalculatedfrom4technicalreplicates.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalSCIDmiceareinjectedsubcutaneouslywith106gliomacells,whicharesuspendedin100μLofa50:50Administration[1]mixtureofgrowthmediaandMatrigel.Oncetumorshavereachedameasurablesize,micearerandomizedintotheindicatedtreatmentgroups.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻(xiàn)?ClinCancerRes.2018Apr1;24(7):1705-1715.?CancerRes.2018Nov15;78(22):6386-6398.?CancerRes.2015Nov15;75(22):4790-802.?FASEBJ.2018Jun7:fj201800547R.?CancerMetab.2019May20;7:4.Seemorecustomervalidationsonwww.MedChemEREFERENCES[1].RohleD,etal.AninhibitorofmutantIDH1delaysgrowthandpromotesdifferentiationofgliomacells.Science.2013May3;340(6132):626-30.[2].MolenaarRJ,etal.RadioprotectionofIDH1-MutatedCancerCellsbytheIDH1-Muta