Medchemexpress/Y-27632 dihydrochloride/HY-10583/50mg

Y-27632dihydrochlorideisanATP-competitiveinhibitorofROCK-IandROCK-II,withK_iof220nMand300nMforROCK-IandROCK-II,respectively.

Ki:220/300nM(ROCK-I/II)^[1]

Y-27632inhibitstheROCKfamilyofkinases100timesmorepotentlythanotherkinasesincludingproteinkinaseC,cAMP-dependentkinaseandmyosinlightchainkinase.Y-27632prolongsthelagtimeanddelaystheappearanceofBrdU-labeledcellsinaconcentration-dependentmanner,delaysofabout1and4harenoticedintheSwiss3T3cellstreatedwith10and100μMY-27632,respectively^[1].Y-27632promotesneuronaldifferentiationofADIposetissue-derivedstemcells(ADSCs).Comparedto1.0and2.5μMY-27632inducedgroups,percentagesofneuroal-likecellsachievedapeakinthe5.0μMY-27632inducedgroup^[2].

Y-27632(5and10mg/kg)significantlyprolongstheonsettimeofmyoclonicjerkswhencomparewithsalinegroup.Y-27632(5and10mg/kg)significantlyprolongstheonsettimeofclonicconvulsionswhencomparewithsalinegroup^[3].TreatmentwithDimethylnitrosamine(DMN)causesasignificantdecreaseinratbodyandliverweight(DMN-Sgroup)comparedwithcontrolanimals(S-Sgroup).OralY27632(30mg/kg)essentiallypreventsthisDMN-inducedratbodyandliverweightloss(DMN-Ygroup)^[4].

• [1].IshizakiT,etal.PharmacologicalpropertiesofY-27632,aspecificinhibitorofrho-associatedkinases.MolPharmacol.2000May;57(5):976-83.

  [2].XueZW,etal.Rho-associatedcoiledkinaseinhibitorY-27632promotesneuronal-likedifferentiationofadulthumanadiposetissue-derivedstemcells.ChinMedJ(Engl).2012Sep;125(18):3332-5.

  [3].InanS,etal.AntiepilepticeffectsoftwoRho-kinaseinhibitors,Y-27632andfasudil,inmice.BrJPharmacol.2008Sep;155(1):44-51.

  [4].TadaS,etal.AselectiveROCKinhibitor,Y27632,preventsdimethylnitrosamine-inducedhepaticfibrosisinrats.JHepatol.2001Apr;34(4):529-36.

RecombinantROCK-I,ROCK-II,PKN,orcitronkinaseisexpressedinHeLacellsasMyc-taggedproteinsbytransfectionusingLipofectamine,andisprecipitatedfromthecelllysatesbytheuseof9E10monoclonalanti-MycantibodycoupledtoGprotein-Sepharose.Recoveredimmunocomplexesareincubatedwithvariousconcentrationsof[³²P]ATPand10mgofhistonetype2assubstratesintheabsenceorpresenceofvariousconcentrationsofeitherY-27632orY-30141at30°Cfor30mininatotalvolumeof30μLofthekinasebuffercontaining50mMHEPES-NaOH,pH7.4,10mMMgCl₂,5mMMnCl₂,0.02%Briji35,and2mMdithiothreitol.PKCaisincubatedwith5μM[³²P]ATPand200μg/mLhistonetype2assubstratesintheabsenceorpresenceofvariousconcentrationsofeitherY-27632orY-30141at30°Cfor10mininakinasebuffercontaining50mMTris-HCl,pH7.5,0.5mMCaCl2,5mMmagnesiumacetate,25μg/mLphosphatidylserine,50ng/mL12-O-tetradecanoylphorbol-13-acetateand0.001%leupeptininatotalvolumeof30μL.Incubationisterminatedbytheadditionof10μLof43Laemmlisamplebuffer.Afterboilingfor5min,themixtureissubjectedtoSDS-polyacrylamidegelelectrophoresisona16%gel.ThegelisstainedwithCoomassieBrilliantBlue,andthendried.Thebandscorrespondingtohistonetype2areexcised,andtheradioactivityismeasured^[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

Y-27632isdissolvedinwaterandstored^[1].

HeLacellsareplatedatadensityof3×10⁴cellsper3.5-cmdish.ThecellsareculturedinDMEMcontaining10%FBSinthepresenceof10mMThymidinefor16h.AfterthecellsarewashedwithDMEMcontaining10%FBS,theyareculturedforanadditional8h,andthen40ng/mLofNocodazoleisadded.After11.5hoftheNocodazoletreatment,variousconcentrationsofY-27632(0-300μM),Y-30141,orvehicleisaddedandthecellsareincubatedforanother30min^[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

Y-27632isdissolvedin0.9%NaCl(saline)(Mice)^[3].
Y-27632isdissolvedinsaline(finalconcentration2%)(Rat)^[4].

Mice^[3]
Male,inbredSwissalbinomice(2-3monthsold)weighing25-30gareused.Miceareinjectedwithasub-convulsivedoseofPTZ(35mg/kg,i.p.)(onMondays,WednesdaysandFridays)ofeachweekforatotalof11injections.AftereachPTZinjection,miceareobservedfor30minandtheoccurrenceofconvulsiveactivityisrecorded.After30min,themicearetheninjectedwitheitherFasudil(25mg/kg,i.p.)orY-27632(5mg/kg,i.p.)andreturnedtotheirhomecagesuntilthenextinjection.ControlmiceforFasudilandY-27632receivessaline.
Rat^[4]
MaleWistarKindArats(200-250g)areused.DMN(1g/mL)isdilutedtentimeswithsaline(finalconcentration1%)and10mg/kgperdayofDMNisinjectedintraperitoneally(i.p.)onthefirst3daysofeachweekfor4weeks.Y27632isgivenorallyonceperdayatadoseof30mg/kgfor4weeksstartingonthedayofthefirstinjectionofDMN.Thedoseof30mg/kgcorrectshypertensioninseveralratmodelswithouttoxicity.Twentyratsarerandomizedintofourexperimentalgroups(n=5ineachgroup)asfollows:(1)S-S(injectionofsalinei.p.andoraladministrationofsaline);(2)S-Y(injectionofsalinei.p.andoraladministrationofY27632);(3)DMN-S(DMNi.p.andoraladministrationofsaline);(4)DMN-Y(DMNi.p.andoraladministrationofY27632).Theratsareweighedeveryweek.Theyaresacrificedattheendofthefourthweekandtheliverisexcised.Inaddition,abloodsampleistakenimmediatelybeforetheratsaresacrificed.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

• [1].IshizakiT,etal.PharmacologicalpropertiesofY-27632,aspecificinhibitorofrho-associatedkinases.MolPharmacol.2000May;57(5):976-83.

  [2].XueZW,etal.Rho-associatedcoiledkinaseinhibitorY-27632promotesneuronal-likedifferentiationofadulthumanadiposetissue-derivedstemcells.ChinMedJ(Engl).2012Sep;125(18):3332-5.

  [3].InanS,etal.AntiepilepticeffectsoftwoRho-kinaseinhibitors,Y-27632andfasudil,inmice.BrJPharmacol.2008Sep;155(1):44-51.

  [4].TadaS,etal.AselectiveROCKinhibitor,Y27632,preventsdimethylnitrosamine-inducedhepaticfibrosisinrats.JHepatol.2001Apr;34(4):529-36.

320.26

C₁₄H₂₃Cl₂N₃O

129830-38-2

RoomtemperatureincontinentalUS;mayvaryelsewhere

DMSO:≥32mg/mL

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