Medchemexpress/LY294002(Synonyms: NSC 697286; SF 1101)/HY-10108/10mg

LY294002isabroad-spectruminhibitorofPI3K,withIC₅₀of0.5/0.57/0.97μMforPI3Kα/δ/β,respectively,alsopotentlyinhibitsCK2withIC₅₀of98nM.

IC50:0.5/0.57/0.97μM(PI3Kα/δ/β)^[1]
IC50:98nM(CK2)^[2]

LY294002(5μM)completelyinhibitsthephosphorylationofPKBInHepG2cells.LY294002(5μM)isalsoshowntoblockinsulin-inducedphosphorylationofPKBSer⁴⁷³inCHO-IRcells^[1].LY294002isalsoapotentinhibitorofCK2(caseinkinase2)withIC₅₀of98nM.LY294002isalsoabletoreducethekinaseactivityofbothisoformsoftheserine/threoninekinasesGSK3αandβ^[2].WhentheCNE-2ZcelllineisculturedinmediumcontainingLY294002(0μM,10μM,25μM,50μM,and75μM)for24hand48h,cellproliferationisremarkablydecreasedinadose-dependentfashion^[3].

TreatmentwithLY294002(i.p.,50mg/kg,75mg/kg)significantlyreducesmeanNPCtumorburdenascomparedwiththecontrolgroup.Treatmentwith10mg/kgor25mg/kgLY294002islesseffectiveindecreasingtumorburden.MeanNPCtumorburdentreatedwithLY294002isremarkablydecreasedinadose-dependentmanner,whereasmeanbodyweightisnoobviousdifferencebetweencontrolandtreatedgroups(LY294002,10mg/kg,25mg/kg,50mg/kg,and75mg/kg)^[3].

• [1].ChaussadeC,etal.EvidenceforfunctionalredundancyofclassIAPI3Kisoformsininsulinsignalling.BiochemJ.2007Jun15;404(3):449-58.

  [2].GharbiSI,etal.ExploringthespecificityofthePI3KfamilyinhibitorLY294002.BiochemJ.2007May15;404(1):15-21.

  [3].JiangH,etal.Phosphatidylinositol3-kinaseinhibitor(LY294002)inducesapoptosisofhumannasopharyngealcarcinomainvitroandinvivo.JExpClinCancerRes.2010Apr22;29:34.

  [4].UedaK,etal.DifferentialeffectsofLY294002andwortmanninonneuronsandvascularendothelialcellsintheratretina.PharmacolRep.2013;65(4):854-62.

PI3KinhibitionbyPI828andLY294002isdeterminedinarADIometricassayusingpurified,recombinantenzymes(classIAandclassIB)with1μMATP.Thekinasereactioniscarriedoutfor1hatroomtemperature(24°C)andisterminatedbyadditionofPBS.IC₅₀valuesaresubsequentlydeterminedusingasigmoidaldose-responsecurvefit(variableslope).CK2andGSK3β(glycogensynthasekinase3β)inhibitionisestablishedbykinaseselectivityscreening.Inhibitor(10μM;PI828andLY294002)istestedagainsttheUpstatepanelofkinasesin10μMATP^[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

LY294002isdissolvedinDMSOandstored,andthendilutedwithappropriatemedia(DMSO0.5%)beforeuse^[3].

HumannasopharyngealcarcinomacelllineCNE-2Zisseededinto96-wellplatesat5000cells/well.Twenty-fourhoursaftercellsareseeded,themediumisremovedandreplacedinthepresenceofLY294002(0μM,10μM,25μM,50μM,and75μM)dissolvedinDMSOorDMSOonlyforanadditional24hand48h.ToavoidanynonspecifictoxiceffectsofDMSOoncellgrowth,DMSOconcentrationsaremaintainedat0.5%inallexperiments.MTTdye(5mg/mL)isaddedtoeachwell.ThereactionisstoppedbytheadditionofDMSO,andopticaldensityismeasuredat490nmonamultiwellplatereader.Backgroundabsorbanceofthemediumintheabsenceofcellsissubtracted.Allsamplesareassayedintriplicate,andthemeanforeachexperimentiscalculated.Resultsareexpressedasapercentageofcontrol,whichisconsideredtobe100%^[3].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

LY294002isdissolvedinvehicle(DMSO).

Mice^[3]
Athymicnudemiceareusedwhentheyare6-8weeks.Micearerandomlydividedintofreeseparatedintofivegroups(n=4mice).Micearehousedinthesameenvironmentwithcontrolledtemperature,humidity,anda12hlight/darkcycle.MiceareinoculatedsubcutaneouslywithCNE-2Zcells(1×10⁶cells/mousein200μLofRPMI-1640)intotheflank.Thetumortakerateis100%.After1week,anintraperitonealinjectionisperformedtothexenograftmicewithdifferentdosageofLY294002(10mg/kg,25mg/kg,50mg/kg,and75mg/kgtwiceweekly(n=4mice),eachgroupfor4weeks.Treatedmicearemonitoredanysigns.Bodyweightandtumorssizearemeasuredtwiceaweek.Tumorsizeismeasuredusingcalipersandtumorvolumeiscalculated(volume=longaxis×shortaxis²).Attheendofthetreatment,allmiceareeuthanized.Onepartoftumortissueisfixedinformalinandembeddedinparaffin,andanotherpartisstoredat-70°C.
Rat^[4]
MaleSprague-Dawleyratsweighing220-240gareanesthetizedbyintraperitoneallyinjectingpentobarbitalsodium(50mg/kg).Theanimalsaredividedinto3groups:NMDA+vehicle(DMSO)(n=46),NMDA+LY294002(50nmol)(n=25),andNMDA+Wortmannin(50nmol)(n=23).EitherLY294002orwortmanninmixedwith200nmolofNMDAinatotalvolumeof5μLisinjectedintothevitreouscavityofoneeye.ThesamevolumeofDMSOisinjectedintothevitreouscavityofthecontralateraleye,whichisusedasacontrol.Theinjectionsareperformedunderamicroscopeusinga32-gaugeneedle,whichisconnectedtoamicrosyringe.Theneedleisinsertedapproximately1mmbehindthecorneallimbus.Damagetoneuronsandbloodvesselsintheretinaisassessedat2and7daysaftertheinjection.TheeffectsoftheintravitrealtreatmentwitheitherLY294002orWortmanninaloneonretinalneuronsandbloodvesselsarealsoexamined.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

• [1].ChaussadeC,etal.EvidenceforfunctionalredundancyofclassIAPI3Kisoformsininsulinsignalling.BiochemJ.2007Jun15;404(3):449-58.

  [2].GharbiSI,etal.ExploringthespecificityofthePI3KfamilyinhibitorLY294002.BiochemJ.2007May15;404(1):15-21.

  [3].JiangH,etal.Phosphatidylinositol3-kinaseinhibitor(LY294002)inducesapoptosisofhumannasopharyngealcarcinomainvitroandinvivo.JExpClinCancerRes.2010Apr22;29:34.

  [4].UedaK,etal.DifferentialeffectsofLY294002andwortmanninonneuronsandvascularendothelialcellsintheratretina.PharmacolRep.2013;65(4):854-62.

307.34

C₁₉H₁₇NO₃

154447-36-6

RoomtemperatureincontinentalUS;mayvaryelsewhere

DMSO:14.9mg/mL

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