ABT-199isahighlypotent,orallybioavailableandBcl-2-selectiveinhibitorwithK_iof<0.01 nm.="">

Ki:<0.01 nm="" (bcl-2),="" 48="" nm="" (bcl-xl),="" 245="" nm="" (bcl-w),="">444nM(Mcl-1)^[1]

ABT-199potentlykillsFL5.12-BCL-2cells(EC₅₀=4nM),ABT-199showsmuchweakeractivityagainstFL5.12-BCL-XLcells(EC₅₀=261nM).ABT-199alsoshowsselectivityincellularmammaliantwo-hybridassays,whereitdisruptsBCL-2-BIMcomplexes(EC₅₀=3nM)butismuchlesseffectiveagainstBCL-XL-BCL-XS(EC₅₀=2.2μM)orMCL-1-NOXAcomplexes^[1].

Afterasingleoraldoseof12.5mgperkgbodyweightinxenograftsderivedfromRS4;11cells(ALL),ABT-199causesamaximaltumorgrowthinhibition(TGI_max)of47%(P<0.001) and="" tumor="" growth="" delay="" (tgd)="" of="" 26%=""><>^[1].Treatmentofestablishedxenografted(amousexenograftmodeloftheT-ALLcelllineLOUCY)tumorswith100mgABT-199/kgfor4daysresultedinasignificantreductionofleukemicburden(P=0.0048)^[2].

• [1].SouersAJ,etal.ABT-199,apotentandselectiveBCL-2inhibitor,achievesantitumoractivitywhilesparingplatelets.NatMed.2013Feb;19(2):202-8.

  [2].PeirsS,etal.ABT-199mediatedinhibitionofBCL-2asanoveltherapeuticstrategyinT-cellacutelymphoblasticleukemia.Blood.2014Dec11;124(25):3738-47.

  [3].BiC,etal.Inhibitionof4EBPphosphorylationmediatesthecytotoxiceffectofmechaNISTictargetofrapamycinkinaseinhibitorsinaggressiveB-celllymphomas.Haematologica.2017Apr;102(4):755-764.

ABT-199isdissolvedinDMSOandstored,andthendilutedwithappropriatemediabeforeuse^[1].

RS4;11cellsareseededat50,000perwellin96-wellplatesandtreatedwithcompoundsdilutedinhalf-logstepsstartingat1μMandendingat0.00005μM.Allotherleukemiaandlymphomacelllinesareseededat15,000-20,000cellsperwellintheappropriatemediumandincubatedwithABT-199orNavitoclaxfor48h.EffectsonproliferationaredeterminedusingCellTiterGloreagent.EC₅₀valuesaredeterminedbynonlinearregressionanalysisoftheconcentration-responsedata.MouseFL5.12-BCL-2andFL5.12-BCL-XLcellsarepropagatedandassessed.Bak^-/-Bax^-/-doubleknockoutmouseembryonicfibroblastsareseededinto96-wellmicrotiterplatesat5,000cellsperwellinDMEMsupplementedwith10%FBS.ABT-199inthesameculturemediumisaddedinhalf-logdilutionsstartingat5μM.Thecellsarethenincubatedat37°C(5%CO₂)for48h,andtheeffectsonproliferationaredeterminedusingCellTiterGloreagent^[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

ABT-199isformulatedin60%phosal50propyleneglycol,30%polyethyleneglycol400,and10%ethanol(Mice)^[2].

Mice^[2]
Nonobesediabetic/severecombinedimmunodeficientγ(NSG)miceareinjectedat6weeksofageinthetailveinwith150µLphosphate-bufferedsalinecontaining5×10⁶luciferase-labeledLOUCYcells.Atregulartimepoints,thebioluminescenceismeasuredusingtheIVISLuminaIIimagingsystem.At6weeks,thecellsareengraftedandthemicearerandomlydividedinto2groups(withanequalnumberofmalesandfemalesinbothgroups),andthetreatmentisstartedonday0.Micearetreatedwith100mgABT-199/kgbodyweightorwithvehicleviaoralgavagefor4consecutivedays.Atdays0,2,and4thebioluminesceneismeasured.Beforeimaging,themiceareinjectedintraperitoneallywith200µLofa15mg/mLfireflyD-luciferinpotassiumsaltsolutionandanesthetizedbyinhalationof5%isoflurane.Themiceareimaged10minutesafterluciferininjection.Thetotalbioluminescencesignalineachmouseiscalculatedviatheregionofinteresttool(totalcounts)intheLivingImagesoftware.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

• [1].SouersAJ,etal.ABT-199,apotentandselectiveBCL-2inhibitor,achievesantitumoractivitywhilesparingplatelets.NatMed.2013Feb;19(2):202-8.

  [2].PeirsS,etal.ABT-199mediatedinhibitionofBCL-2asanoveltherapeuticstrategyinT-cellacutelymphoblasticleukemia.Blood.2014Dec11;124(25):3738-47.

  [3].BiC,etal.Inhibitionof4EBPphosphorylationmediatesthecytotoxiceffectofmechanistictargetofrapamycinkinaseinhibitorsinaggressiveB-celllymphomas.Haematologica.2017Apr;102(4):755-764.

868.44

C₄₅H₅₀ClN₇O₇S

1257044-40-8

RoomtemperatureincontinentalUS;mayvaryelsewhere

DMSO:≥51mg/mL

ABT-199ispreparedin60%phosal50PG,30%PEG400and10%ethanol^[3].

*"<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">

• [1].SouersAJ,etal.ABT-199,apotentandselectiveBCL-2inhibitor,achievesantitumoractivitywhilesparingplatelets.NatMed.2013Feb;19(2):202-8.

  [2].PeirsS,etal.ABT-199mediatedinhibitionofBCL-2asanoveltherapeuticstrategyinT-cellacutelymphoblasticleukemia.Blood.2014Dec11;124(25):3738-47.

  [3].BiC,etal.Inhibitionof4EBPphosphorylationmediatesthecytotoxiceffectofmechanistictargetofrapamycinkinaseinhibitorsinaggressiveB-celllymphomas.Haematologica.2017Apr;102(4):755-764.