SimvastatinisacompetitiveinhibitorofHMG-CoAreductasewithK_dof0.1-0.2nM.

Ki:0.1-0.2nM(HMG-CoAreductase)

Priortouseincellassays,simvastatinneedstobeactivatedbyNaOHinEtOHtreatment.SimvastatininhibitscholesterolsynthesisinmouseL-Mcell(fibroblast),ratH4IIEcell(liver),andhumanHepG2cell(liver)withIC₅₀of19.3nM,13.3nMand15.6nM,respectively^[1].Simvastatintreatmentleadstoadose-dependentincreaseinserine473phosphorylationofAktwithin30minutes,withmaximalphosphorylationoccurringat1.0µM.Simvastatin(1.0μM)enhancesphosphorylationoftheendogenousAktsubstrateendothelialnitricoxidesynthase(eNOS),inhibitsserum-freemediaundergoapoptosisandacceleratesvascularstructureformation^[2].Simvastatindisplaysanti-inflammatoryeffectsinvitro.Simvastatin(10μM)reducesanti-CD3/anti-CD28antibody-stimulatedproliferationofPB-derivedmononuclearcellsandsynovialfluidcellsfromrheumatoidarthritisblood,aswellasIFN-γrelease.Simvastatin(10μM)suppressescell-mediatedmacrophageTNF-γreleaseinducedviacognateinteractionsbyappr30%^[3].

SimvastatinorallyadmiNISTrationinhibitstheconversionofrADIolabeledacetatetocholesterolwithIC₅₀of0.2mg/kg^[1].Simvastatin(4mg/day)orallyadministrationfor13weekstorabbitsfedanatherogencicholesterol-richdiet,returnsthecholesterol-inducedincreasesintotalcholesterol,LDL-cholesterolandHDL-cholesteroltonormallevel^[4]. Simvastatin(6mg/kg)producesanincreaseinLDLreceptor-dependentbindingandincreasesthenumberofhepaticLDLreceptorsinrabbitsfedadietcontaining0.25%cholesterol^[5].Simvastatininfluencesinflammationindependentofitseffectonplasmacholesterollevel.Incynomolgusmonkeysconsumedanatherogenicdiet,Simvastatin(20mg/kg/day)inducesa1.3-foldlessmacrophagecontentinlesions,and2-foldlessvascularcelladhesionmolecule-1,interleukin-1beta,andtissuefactorexpression,companiedbya2.1-foldincreasesinlesionalsmoothmusclecellandcollagencontent^[6].

• [1].Slater,E.E.,etal.MechanismofactionandBIOLOGicalprofileofHMGCoAreductaseinhibitors.Anewtherapeuticalternative.Drugs,1988.36Suppl3:p.72-82.

  [2].Kureishi,Y.,etal.TheHMG-CoAreductaseinhibitorsimvastatinactivatestheproteinkinaseAktandpromotesangiogenesisinnormocholesterolemicanimals.NatMed,2000.6(9):p.1004-10.

  [3].LeungBP,etal.Anovelanti-inflammatoryroleforsimvastatinininflammatoryarthritis.JImmunol.2003Feb1;170(3):1524-30.

  [4].KobayashiM,etal.PreventiveeffectofMK-733(simvastatin),aninhibitorofHMG-CoAreductase,onhypercholesterolemiaandatherosclerosisinducedbycholesterolfeedinginrabbits.JpnJPharmacol.1989Jan;49(1):125-33.

  [5].IshidaF,etal.Comparativeeffectsofsimvastatin(MK-733)andpravastatin(CS-514)onhypercholesterolemiainducedbycholesterolfeedinginrabbits.BiochimBiophysActa.1990Feb23;1042(3):365-73.

  [6].SukhovaGK,etal.Statinsreduceinflammationinatheromaofnonhumanprimatesindependentofeffectsonserumcholesterol.ArteriosclerThrombVascBiol.2002Sep1;22(9):1452-8.

  [7].ZhongzhongLiu,etal.PretreatmentDonorsafterCirculatoryDeathwithSimvastatinAlleviatesLiverIschemiaReperfusionInjuryThroughaKLF2-dependentMechanisminRat.OxidMedCellLongev.24July2017.

ForassessmentofAktproteinkinaseactivityinvitro,substrate(2μghistoneH2Bor25μgeNOSpeptide)isincubatedwithAktimmunoprecipitatedfromcelllysateusinggoatpolyclonalanti-Akt1antibody.KinasereactionsareinitiatedfollowingtheadditionofreactioncomponentstoafinalconcentrationofATP(50μM)containing10μCiof³²P-γATP,dithiotreitol(1mM),HEPESbuffer(20mM,pH7.4),MnCl₂(10mM),MgCl₂(10mM).Afterincubationfor30minat30°C,phosphorylatedhistoneH2BisvisualizedafterSDS-PAGE(15%)andautoradiography.Toestimatetheextentof³²PincorporationintoeNOSpeptides,eachreactionmixtureismeasuredbyspottingontophosphocellulosediscfilterandtheamountofphosphateincorporatedismeasuredbyCerenkovcounting.Thewild-typepeptidesequenceis1174-RIRTQSFSLQERHLRGAVPWA-1194,andthemutanteNOSpeptideisidenticalexceptthatserine1179issubstitutedbyalanine.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

Thirty-nineadultmalecynomolgusmonkeysareinitiallyfedamoderatelyatherogenicdietcontaining0.28mgcholesterolpercalorieofdiet,with16.7%fromprotein,45.1%fromlipids,and38.1%fromcarbohydrates.Afterconsumingtheatherogenicdietfor3months,themonkeysaredividedinto3groups(n=13each)thatareequivalentintheirtotalplasmacholesterol(TPC),LDL-C,andHDLcholesterol(HDL-C)concentrations;thesegroupsconsumetheatherogenicdietandreceivestatin(orcontrol)treatmentforanadditional15months.Controlmonkeysarefedtheatherogenicdietwithnoaddedstatins.Prava-treatedmonkeyshave40mgPrava/kgbodywtperdayaddedtotheatherogenicdiet.Simvastatin(Simva)-treatedmonkeysconsumed20mgSimva/kgbodywtperday.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

• [1].Slater,E.E.,etal.MechanismofactionandbiologicalprofileofHMGCoAreductaseinhibitors.Anewtherapeuticalternative.Drugs,1988.36Suppl3:p.72-82.

  [2].Kureishi,Y.,etal.TheHMG-CoAreductaseinhibitorsimvastatinactivatestheproteinkinaseAktandpromotesangiogenesisinnormocholesterolemicanimals.NatMed,2000.6(9):p.1004-10.

  [3].LeungBP,etal.Anovelanti-inflammatoryroleforsimvastatinininflammatoryarthritis.JImmunol.2003Feb1;170(3):1524-30.

  [4].KobayashiM,etal.PreventiveeffectofMK-733(simvastatin),aninhibitorofHMG-CoAreductase,onhypercholesterolemiaandatherosclerosisinducedbycholesterolfeedinginrabbits.JpnJPharmacol.1989Jan;49(1):125-33.

  [5].IshidaF,etal.Comparativeeffectsofsimvastatin(MK-733)andpravastatin(CS-514)onhypercholesterolemiainducedbycholesterolfeedinginrabbits.BiochimBiophysActa.1990Feb23;1042(3):365-73.

  [6].SukhovaGK,etal.Statinsreduceinflammationinatheromaofnonhumanprimatesindependentofeffectsonserumcholesterol.ArteriosclerThrombVascBiol.2002Sep1;22(9):1452-8.

  [7].ZhongzhongLiu,etal.PretreatmentDonorsafterCirculatoryDeathwithSimvastatinAlleviatesLiverIschemiaReperfusionInjuryThroughaKLF2-dependentMechanisminRat.OxidMedCellLongev.24July2017.

418.57

C₂₅H₃₈O₅

79902-63-9

RoomtemperatureincontinentalUS;mayvaryelsewhere

10mMinDMSO

Simvastatinispreparedinvehicle(DMSO0.1%)^[7].

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

• [1].Slater,E.E.,etal.MechanismofactionandbiologicalprofileofHMGCoAreductaseinhibitors.Anewtherapeuticalternative.Drugs,1988.36Suppl3:p.72-82.

  [2].Kureishi,Y.,etal.TheHMG-CoAreductaseinhibitorsimvastatinactivatestheproteinkinaseAktandpromotesangiogenesisinnormocholesterolemicanimals.NatMed,2000.6(9):p.1004-10.

  [3].LeungBP,etal.Anovelanti-inflammatoryroleforsimvastatinininflammatoryarthritis.JImmunol.2003Feb1;170(3):1524-30.

  [4].KobayashiM,etal.PreventiveeffectofMK-733(simvastatin),aninhibitorofHMG-CoAreductase,onhypercholesterolemiaandatherosclerosisinducedbycholesterolfeedinginrabbits.JpnJPharmacol.1989Jan;49(1):125-33.

  [5].IshidaF,etal.Comparativeeffectsofsimvastatin(MK-733)andpravastatin(CS-514)onhypercholesterolemiainducedbycholesterolfeedinginrabbits.BiochimBiophysActa.1990Feb23;1042(3):365-73.

  [6].SukhovaGK,etal.Statinsreduceinflammationinatheromaofnonhumanprimatesindependentofeffectsonserumcholesterol.ArteriosclerThrombVascBiol.2002Sep1;22(9):1452-8.

  [7].ZhongzhongLiu,etal.PretreatmentDonorsafterCirculatoryDeathwithSimvastatinAlleviatesLiverIschemiaReperfusionInjuryThroughaKLF2-dependentMechanisminRat.OxidMedCellLongev.24July2017.