Medchemexpress/(+)-JQ-1(Synonyms: JQ1)/HY-13030/10mM*1mL in DMSO

(+)-JQ-1isaBETbromodomaininhibitor,withIC₅₀of77nM/33nMforthefirstandsecondbromodomain(BRD4(1/2)).

IC50:77/33nM(BRD4(1/2))^[1]

(+)-JQ1representsapotent,highlyspecificandKaccompetitiveinhibitorfortheBETfamilyofbromodomains.(+)-JQ1(100nM,48h)promptssquamousdifferentiationexhibitedbycellspindling,flatteningandincreasedexpressionofkeratin.(+)-JQ1(250nM)inducesrapidexpressionofkeratinintreatedNMC797cellscomparedto(-)-JQ1(250nM)andvehiclecontrols,asdeterminedbyquantitativeimmunohistochemistry.(+)-JQ1(250nM)elicitsatime-dependentinductionofstrong(3+)keratinstainingoftreatedNMC797cells,comparedto(-)-JQ1(250nM)^[1].(+)-JQ1isapotentthienodiazepineinhibitor(K_d=90nM)oftheBETfamilycoactivatorproteinBRD4,whichisimplicatedinthepathogenesisofcancerviatranscriptionalcontroloftheMYConcogene.Dose-rangingstudiesof(+)-JQ1demonstratespotentinhibitionofH4Kac4bindingwithaIC50valueof10nMformurineBRDT(1)and11nMforhumanBRDT(1)^[2].

Pharmacokineticstudiesof(+)-JQ1areperformedinCD1micefollowingintravenousandoraladmiNISTration.Meanplasmaconcentration-timeprofilesof(+)-JQ1afterintravenousdosing(5mg/kg).Thepharmacokineticparametersforintravenous(+)-JQ1demonstrateexcellentdrugexposure(AUC=2090hr*ng/mL)andanapproximatelyonehourhalf-life(T1/2).Meanplasmaconcentration-timeprofilesof(+)-JQ1afteroraldosing(10mg/kg).Thepharmacokineticparametersfororal(+)-JQ1demonstrateexcellentoralbioavailABIlity(F=49%),peakplasmaconcentration(C_max=1180ng/mL)anddrugexposure(AUC=2090hr*ng/mL)^[1].

• [1].FilippakopoulosP,etal.SelectiveinhibitionofBETbromodomains.Nature.2010Dec23;468(7327):1067-73.

  [2].MatzukMM,etal.Small-moleculeinhibitionofBRDTformalecontraception.Cell.2012Aug17;150(4):673-84.

  [3].PeirsS,etal.TargetingBETproteinsimprovesthetherapeuticefficacyofBCL-2inhibitioninT-cellacutelymphoblasticleukemia.Leukemia.2017Feb3.

  [4].TögelL,etal.DualTargetingofBromodomainandExtraterminalDomainProteins,andWNTorMAPKSignaling,Inhibitsc-MYCExpressionandProliferationofColorectalCancerCells.MolCancerTher.2016Jun;15(6):1217-26.

  [5].SahniJM,etal.BromodomainandExtraterminalProteinInhibitionBlocksGrowthofTriple-negativeBreastCancersthroughtheSuppressionofAuroraKinases.JBiolChem.2016Nov4;291(45):23756-23768.

  [6].NakamuraY,etal.Targetingofsuper-enhancersandmutantBRAFcansuppressgrowthofBRAF-mutantcoloncancercellsviarepressionofMAPKsignalingpathway.CancerLett.2017Aug28;402:100-109.

  [7].BhattacharyyaS,etal.Alteredhydroxymethylationisseenatregulatoryregionsinpancreaticcancerandregulatesoncogenicpathways.GenomeRes.2017Nov;27(11):1830-1842.

(+)-JQ1isdissolvedinDMSOandstored,andthendilutedwithappropriatemedia(DMSO0.025%)beforeuse^[1].

NUTmidlinecarcinomapatientcelllines(797and11060)areplatedinT-25flasksandgrowninDMEM(797)orRPMI(11060)containing10%fetalbovineserumand1%Penicillin/Streptomycin.Cellsaretreatedwitheither250nM(+)-JQ1,250nM(-)-JQ1ortheequivalentvolumeofDMSO(0.025%).Atthedesiredtimepoint,2×10⁶cellsarespunat500×gfor5minutesat4°CandwashedwithPBS.PelletsareresUSPendedin1mLofcoldPBSandaddeddropwisewhilegentlyvortexingto9mL70%ethanolina15mLpolypropylenecentrifugetube.Fixedcellsarethenfrozenat-20°Covernight.Thenextday,cellsarecentrifugedat500×gfor10minutesat4°Candwashedwith3mLofcoldPBS.Cellsareresuspendedin500μLofpropidiumiodidestainingsolution(0.2mg/mLRNAseA,0.02mg/mLpropidiumiodide,0.1%Triton-XinPBS)andincubatedfor20minutesat37°C.SamplesarethentransferredtoiceandanalyzedonaBDFACSCantoII.HistogramsaregeneratedandcellcycleanalysisisperformedusingFlowJoflowcytometryanalysissoftware^[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

(+)-JQ1isformulatedforintravenousinjectionin10%DMSOand10%HP-β-CD(Mice)^[1].
(+)-JQ1isformulatedinsaline,10%2-Hydroxypropyl-β-cyclodextrinand10%DMSO(Rat)^[2].

Mice^[1]
MaleCD1mice(24-29g)aretreatedwithasingledoseof(+)-JQ1at5mg/kgforintravenoustailveininjectionstudiesand10mg/kgfororalgavagestudies.Approximately150μLofbloodaretakenfromanimalsbyretro-orbitalpunctureunderanesthesiawithIsofluraneintoEDTAtubesatpre-specifiedtimeintervals:0.033,0.083,0.25,0.5,1,2,4,5,8and24hours.Threeanimalsareanalyzedpertimepoint.Bloodsamplesareputoniceandcentrifugedtoobtainplasmasamples(2000×g,5minunder4°C)within15minutespost-sampling.Plasmasamplesarestoredatapproximately-70°Cuntilanalysisisperformed.Miceareprovidedfreeaccesstofoodandwaterthroughoutthestudy.
Rat^[2]
AdultmaleSprague-Dawleyratsaretreatedwithvehicleor(+)-JQ1(10mg/kg).TreatmentisadministeredIPat1/100bodymass.Ratsarecheckedtwice-dailyformortalityandweighedondays1,3,7,14,and21.Thetreatmentregimenutilized4daysof50mg/kgJQ1administereddailywhichisdecreasedto10mg/kgtwicedailyfortheremainderofthestudyduetotheappearanceofadverseeffectsinasubsetofanimals.Forallanimalscompleting3weeksoftreatment,testismass,spermmotility,andspermcountsaredeterminedasdescribedformousestudies.Inbrief,testesarefixedinBouin’sandpreparedforhistology.TheotherhalfismincedinwarmM16bufferandusedforspermcountsandmotilitystudies.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

• [1].FilippakopoulosP,etal.SelectiveinhibitionofBETbromodomains.Nature.2010Dec23;468(7327):1067-73.

  [2].MatzukMM,etal.Small-moleculeinhibitionofBRDTformalecontraception.Cell.2012Aug17;150(4):673-84.

  [3].PeirsS,etal.TargetingBETproteinsimprovesthetherapeuticefficacyofBCL-2inhibitioninT-cellacutelymphoblasticleukemia.Leukemia.2017Feb3.

  [4].TögelL,etal.DualTargetingofBromodomainandExtraterminalDomainProteins,andWNTorMAPKSignaling,Inhibitsc-MYCExpressionandProliferationofColorectalCancerCells.MolCancerTher.2016Jun;15(6):1217-26.

  [5].SahniJM,etal.BromodomainandExtraterminalProteinInhibitionBlocksGrowthofTriple-negativeBreastCancersthroughtheSuppressionofAuroraKinases.JBiolChem.2016Nov4;291(45):23756-23768.

  [6].NakamuraY,etal.Targetingofsuper-enhancersandmutantBRAFcansuppressgrowthofBRAF-mutantcoloncancercellsviarepressionofMAPKsignalingpathway.CancerLett.2017Aug28;402:100-109.

  [7].BhattacharyyaS,etal.Alteredhydroxymethylationisseenatregulatoryregionsinpancreaticcancerandregulatesoncogenicpathways.GenomeRes.2017Nov;27(11):1830-1842.

456.99

C₂₃H₂₅ClN₄O₂S

1268524-70-4

RoomtemperatureincontinentalUS;mayvaryelsewhere

DMSO:≥45mg/mL

(+)-JQ-1(JQ1)isformulatedin10%DMSOand90%ofa10%2-hydroxypropyl-β-cyclodextrinsolution^[3].
(+)-JQ-1(JQ1)ispreparedinvehicle(20%hydroxypropyl-β-cyclodextrin,5%DMSO,0.2%Tween-80insaline)^[4].
(+)-JQ-1(JQ1)ispreparedinvehicle(1:1propyleneglycol:water)^[5].
(+)-JQ-1(JQ1)ispreparedinvehicle(5%DMSOin10%2-hydroxypropyl-ß-cyclodextrinsolution)^[6].
(+)-JQ-1ispreparedinvehicle(10%hydroxypropylβ-cyclodextrininwater)^[7].

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

• [1].FilippakopoulosP,etal.SelectiveinhibitionofBETbromodomains.Nature.2010Dec23;468(7327):1067-73.

  [2].MatzukMM,etal.Small-moleculeinhibitionofBRDTformalecontraception.Cell.2012Aug17;150(4):673-84.

  [3].PeirsS,etal.TargetingBETproteinsimprovesthetherapeuticefficacyofBCL-2inhibitioninT-cellacutelymphoblasticleukemia.Leukemia.2017Feb3.

  [4].TögelL,etal.DualTargetingofBromodomainandExtraterminalDomainProteins,andWNTorMAPKSignaling,Inhibitsc-MYCExpressionandProliferationofColorectalCancerCells.MolCancerTher.2016Jun;15(6):1217-26.

  [5].SahniJM,etal.BromodomainandExtraterminalProteinInhibitionBlocksGrowthofTriple-negativeBreastCancersthroughtheSuppressionofAuroraKinases.JBiolChem.2016Nov4;291(45):23756-23768.

  [6].NakamuraY,etal.Targetingofsuper-enhancersandmutantBRAFcansuppressgrowthofBRAF-mutantcoloncancercellsviarepressionofMAPKsignalingpathway.CancerLett.2017Aug28;402:100-109.

  [7].BhattacharyyaS,etal.Alteredhydroxymethylationisseenatregulatoryregionsinpancreaticcancerandregulatesoncogenicpathways.GenomeRes.2017Nov;27(11):1830-1842.