维甲酸自乳化给药系统的体外溶出及比格犬药物动力学研究

维甲酸自乳化给药系统的体外溶出及比格犬药物动力学研究 维甲酸自乳化给药系统的体外溶出及比格 犬药物动力学研究 JournalofChinesePharmaceuticalSciences2005,14(2):105—109105 InvitroDissolutionandPharmacokineticsinBeagleDogsofa Self-EmulsifyingFormulationofTretinoin QUANDong—qinandXUGui—xia (InstituteofPhamuwolog?andToxicolog?,4cadem)ofMilita0Medica1.%iences,Beijing100850,China) Abstract:AimInvitrodissolutiontestandpharmacokineticsinbeagleclogswereconductedto assessthefommlationoftretinoininself-emulsifyingsystems.MethodsTheconcentrationsoftretinoin weredeterminedbyHPLC.Aerossoverstudywasperformedinfourfastingbeagledogswiththefommla— tionofself-emulsifyingsystemsandcommercialcapsules.ResulIsTheresultsshowedthatthedissolu— tionratein15minoftretinoininself-emulsifyingsystemswashierthan80r/rwhilethatofthecomnler— cialcapsuleswaslowerthan5%.rheareaundertheplasmaconcentration— timecuiwe(AUC)oftheself- emulsifyingformulationwassignificantlyhigherandCwasapproximatelytwotimes~'eaterthanthoseof conunercialcapsule,respectively.Inaddition,thetimetakentoreachpeakwasshorter(2hto1.25h) forself-emulsifyingfommlationoftretinoin.ConclusionIeself— emulsifyingdrugdelivery. systemsCall significantlyincreasetretinoinvitrodissolutionandvivoabsorption. Keywords:self-emulsifyingsystenrs;tretinoin;bioavailability;dissolution CLCnumber:R969.1Documentcode:AArticleID:1O03—1057(2005)2—105—05 Introduction Tretinoinorall—tpans—retinoicacid(R4),ametalMite ofvitaminA,iscapableofregulatingepithelialcellgrowth anddifferentiation,andsebumD~tuction,especiallyfor theirpotentialincancerchemoprophylaxisandtherapy. Duetothelowsolubilityinwater,oraldosageformsofRA suchastabletsandcapsulesinthenkarkethavelowbioavail— ability.FortheclinicaluseofRAincancertreatment.the pooraqueoussolubilityofRAshouldbeimproved.Several meansforimprovingitsaqueoussolubilityhavel:~enstudied suchasliposomes,SLNformulation,andinclusion withcyclodextrin.Self-emulsifyingdrugdeliverysys— tents(SEDDS)areisotropicmixturesofoils,surfactants anddrugs.Uponmildtationfollowedbydilution inaqueousmedia,suchasgastrointestinal(GI)fluids, thesesystenrscanformfineoil—in—water(O/W)emulsions ormicroemulsions(SMEDDS).n1esmalleroildroplets providealargeinterracialareaandtherebypro— Reeeiveeldate:2(x)4—09.18 Correspondingauthor:Tel86—10—66931636, E—mailw出qdq@s0hu.(!onl motetherapidreleaseofthedrug?.Self-enmlsifying fommlationsreadilyspreadintheGItract,andthediges— tiremotilityofthestomachandtheintestineprovidethe agitationnecessaryforself—emulsification".SEDDSare morestablethanemulsions.Forlipophilicdrugcoin— poundsthatexhibitdissolutionrate—limitedabsorption, SEDDSmayofferanimprovementintherateandextentof absorption?,andresuItin171orereprcxtuciblebloed— timeprofiles.Theabsorptionimprovementisincreasedby dissolutionrateofthedrug.Theobjectiveofpresentstudy wastodevelopandcharacterizetheself-emulsifyingdrug deliverysvstems(SEDDS)ofRA. MaterialsandMethods Materialsandapparatus SEDDSofRAwereprepa~bythisl'a~ratory(10n unit),RAcapsuleswerepumtuasedfronlChongqingHtuC,ang PharmaceuticalCong~any(10rag/unit,batchNo. 21301(NI2).Beagledogs(13—15kg,male)werefromAni— realCenterofAcademyofMilitaryMedicaliences. Highspeedcentrifuge(HemlleZ200A)wasnladein German,HPLC(HitachiL一7100)wasmadeinJapan, vortexnlixer(QL-901)wasmadeinShallghai,China, UV一160Av/s—uhravioletspectrophotometer(Shimazu)was 106JournalofChinesePharmaceuticalSciences2005,14(2):105—109 madeinJapan:drugdissolutiontester(RC一3B)wasmade inTianjinUniversityFactory,China. PreparationofSEDDSofRA Theformulationusedfor12vitroandinvivoevalu— ationwasdevelopedafteroilsandsurfactantswere screenedaccordingtothesolubilityofRA.Triglycerides (MIGLYOL812N)oilwasusedasoilphase.Tween80 and1.2.propanolwereusedassurfaetantandcosui~ac— rant.respectively.RA.vitamineEandBHTwereadded tothemixtureofoil/surfactant(3/2,W/W)and1,2一pro— panolbytheaidofstirringfor30minuntilaclearsolu— tionwasachieved.Onegramofthesolutioncontained20 mgRA.I1heSEDDSsoftcapsuleswerepreparedbyfilling with0.50gsolution,equivalentto10n唱ofRApercap— slde. /nvitrodissolutiontest DissolutionwasmeasuredaccordingtoMethodIin ChinesePharmacolx~eia(CPaX~.part?).rn1eIn~un3was de-ionizedwaterandmaintainedat37?0.5cc.rI11ecom— rf~reialcapsulesandself-emulsifyingformulationsofKA_in softcapsuleswerepositionedinthebasketatt=0min. Samplesof5mLwerewithdrawnat5,10,15,20,30,45, 60.and90min.andcentrifugedfor10minat10000r? Inl'n, ,andthenthesupematantwasfiheredwitha0.22p.m filter.Aftersampling,5mLwaterwassimultaneouslyadded tothedissolutionflask.Dissolutiontestforcommercialcap— suleswasperformedinthemediUlTlof0.02%Tween80solu— tion(500n也).Eachdissolutionwasrepeatedforthree times.rI11eaccun~tlatedamountofreleaseddrugateach samplingpointwascorrectedwitI1thevolumeofthedissolu— tionhi.urn.n1econcentrationofRAineachsamplewas determinedbyI-IPI~.rI11emobilephaseconsistedofaceto— I1ile-0.1%ana~niumacetatesolution(90:10,V/V), theflowrate"~gdS1IT1I?min.andthecolumnwasKro— macil—CI8(4.6nm1×150iY~ll,5tan).Analiquotof20 wasinjectedonto}眦.anddeterminationwavelengthwas 340Bil1.rheretentiontimeofRAwasaround9min.1he linearrangeoftretinoinwasfrom0.5mLto10O. mL,.rI11ecalibrationequationwasC=一9018.7+ 172039A(n=8),whereCistheconcentrationofRA.and Aisthepeak—arP_.floftretinoin.I11ecol'l~lationcoefficient was0.9998.rI11edetectionlimitoftheassaywasabout3lag atasign-to-noiseratioof3:1.Precisiondatawereobtained byrepeatedanalysisofsamplesinconcentrationsof5.0,10, and12/xg'n1I一_..Win-dayRsD(n=5)were1.40%. 1.30%,and0.90%,respectively,andbetween.clayRSD% (n=3)were2.2O%,1.01%,and2.01%,respectively. 3herecoverieswere100.3%,99.5%,and102.5%(n= 5),respectively. /nvivostudyinbeagledogs Theanimalswereadministeredfromcrossoverdesign asshowninTable1withawashoutperiodof1week.M— teranovernight(12h)fasting,fourbeagledogswere orallyadministeredwithasingledoseof10mg.Awas givencommercialcapsules;BwasVenself-emulsifying formulations.Waterwasallowedat2handfoodat6h post—dosing.Approximately3mLbloodsamplesweretak— enfromcephalicveincathetersat0(priortodosing), 0.25,0.5,0.75,1.0,1.25,1.5,2.0,2.5,3.0, 4.0,6.0,and8.0h.Plasmawasseparatedbycentri— fugationat3500r.min,for20min.andfrozenat一20~C untilanalysis Table1Drugadministrationtobeagledogs Administrationperiod?min~inun~oer l234 MeasurementofplasmaRAconcentration Analiquotof500ttLphosphatebuffersolution (pH7.0)containing10Lofnimodipine(0.5mg. mL,,asanintemalstandard)wasaddedto0.5mL plasmasolution,andthen2mLmixtureofether.acetate ester(50:50,V/V)wasadded.Thesampleswereshak— enfor5minbyvortexmixerandcentrifugedat3000r? minf0r5min.I1hesupemantwasseparatedandevapo— ratedtodrynessat50ccundernitrogen.Residueswere dissolvedin50HLmobilephaseandassayedbythefol— lowingmodified瑚PLCmethod:mobilephaseconsistedof acetonirile-methanol—water一10%arnnloniumacetatesolu— tion(60:16:20:4,V/V),andflowratewas1n1IJ? nll'n, .I1hecolumnandthedetectionwavelengthwerethe saIneasthosevitrodissolutiontests.I1hestandard cu1weforRArangedfrom50to10000ng?n1IJ,.I1he calibrationequationwasC=一9.224+1193.3A(n= 5),wereCistheconcentrationofRAinplasma.andA isthepeak—arearatioofRAtointemalstandard.I1hecor— relationcoefficientwas0.9984.I1hedetectionlimitof theassaywasabout0.4ngatasign-to-noiseratioof3:1. Precisiondatawereobtainedbyrepeatedanalysisofplas. masamplesinconcentrationsof50.5oO.and10000ng? mL,,within-dayRSD(n=5)were6.33%,8.51%. QUANI~g-clin,et:Inv/troDissolutionandPharmacokineficsinBeag0e 【)0ofaSelf-Emul~iOingFom~lationoflbetin—oin107 and5.96%,between—dayRSD(n=3)were7.44%, 9.37%,and9.3%;themethodl~coveries(n=5)were 85.3%,93.0%,and103.9%,respectively. 3hea瑚undertheplasmaconcentration—timecurve (AUCot)wascalculatedfromobserveddatapointsbyt}1e hneartrapezoidalrule.AUCowascalculatedasAbCot+ Ct/K,wereKwaseliminationconstantobtainedby 3p87program.Themaximumplasmaconcentration (C一)andthecorrespondingpeaktime(f,)wereob— servedvalues.I'heparametersfordiflfPrentformulations werecomparedbyFtestusingSARSsoftware. Results Invitrodissolutiontest Dissolutionresultsshowedthatnearly80%ofRA dissolvedfromtheserf-emulsifyingformulationswithin15 minasshowninFigure1.andonly5%ofRAwasre— leasedfromthecornmercialcapsuleswithinthesainetime period,demonstratingthattheserf-emulsifyingsystenrs couldsignificantlyincreaseRAdissolutionrate. 0102030405060708090l00 Cmin Figure1DissolutionprofilesofRApreparations/nv/tro(n=6) ~Self-emulsifyingformulationinsoftcapsules~Commercialcapsules Pharmacokineticsinbeagledogs MeanplasmaprofilesofRAai'eshowninFigure2. ThepharmacokineticparametersarelistedinTable2. TheresultsshowedthattheAUCff0rRAadministered asserf-emulsifyingsystenrs67%greaterthanthatfor commercialcapsules.ThemeanvalueofCforserf- emulsifyingsystenrswasnearlytwotimesgreaterthanthe C—obtainedwiththesallledoseofRAadministeredas marketedcapsules(1531.71ng?mL,793.18ng? mL).fforserf-emulsifyingformulationswasearlier thanthatofthemarketedproducts(1.25hDersll~2.0h). TheAUC(o)(P<0.O1),AUC(((P<0.01),and C呲(P<0.01)betweenthetwoformulationsweresig— nificantlydifferent. 0246 t/min Figure2Plasmaconcentration-timeprofilesofRApreparations inb~gledogs(n=4).~Self-emulsifyingfomlulationinsoftcap— sules~Commercialcapsules Table2Pharmacokineticparametersoftretinoinpreparations inb~g[edogs(n=4) Discussion ToprepareSEDDSofRA.severaloilsandsuffacta— ntswereused.AccordingtothesolubilityofRA,medium chaintriglyceride(MICLY0L812N)andTween80were selected,buttheycouldnotdissolvemarkedanlountsof RAuntillargeamounts(>80%)ofTween80wasadd— ed.1,2一propanolshouldbeaddedasacosuffactant.and only20%(W/W)Tween80inthefommlationCOUldDro— duceemulsionswithadropletsizebetween300and500 rm1. Tretinoinwasrapidlyisomerisedanddestroyedinthe presenceoflightandoxidant;thereforeconsiderableat— tentionmustbeVentothecollection,handlingandstor— ageofbiologicalsamples.Allsamplesshouldcarefully ?????????0蚴?瑚?蛐?枷瑚0 加???们加0 一一口0一善10??弓J0 1O8JournalofChinesePharmaceuticalSciences2005,14(2):105—109 avoidlightandsamplesshouldbehandledassoonaspos— sible.T}lestabilityofRAsampleswasinvestigatedinour experimentalcondition;theresultsshowedthattheywere stablewithin3d,andthussampleassayshouldbecorn— pletedwithin3d(datanotshown). Fornvitrodissolutionstudies.whenwaterwithout Tween80wasusedforthedissolutionmediumofcommer— cialcapsules.wefoundthatyellowRApowderfloatedon thesurface,andRAwasreleasedsolittlethatthedrug couldnotbedetectedbyHPLC.Itseemedthattheaddit— ionofTween80mightincreasethewettingof1La_.Hence, theeffectofincreasedwettingofRAofTween80in SEDDSformulationmaybeduetotheincFeaseddissolu— tionrateofthedrug,thusleadingtoanimprovedbio— availability. WaterwithaddedTween80(0.02%,W/V)for measuringthedissolutionofcommercialcapsulewasused inordertomaintainthesalllefinalconcentrationof Tween80inthemeditm1. Conclusion Self-emulsifyingdrugdeliverysystemsignificantly increasedinvitrodissolutionandinvivobioavailabilityof tretinoin. Acknowledgements WewouldliketoexpressourgratitudetoProf.R Jinxiuforconsultationinplasmaconcentrationmeasur— ment.ThisworkwassupportedbyNationalMedical FoundationofPIJAinChina. [1jChivotM.Retinoid~empyforacne:acomparativereview lJj.AminDermatol,2005,6(1):13—19. [2jDaiX,YamasakiK,ShirakataY,eta1.刈一trans—retinoic acidinducesinterleukin一8v/athenuclearfactor-kappaBand p38mitogen?-activatedproteinkinasepathwaysinnormalhu?? mallkemtinoeytes[JJ.J,,,Demur>z,2004,123(6): 10r78—1085. [3jParthasarathyR,GilbertB,MehtaK.Aerosoldeliveryofli— posomalall—trans—retinoicacidtothelungs[J].CancerChe— motherPharmacol,1994,43:277—283. 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QUANDong-qin,d:/nv/troDissolutionandHaan~mkineticsinBeag0eofaErr山FomLtlafionofT~tin—oin109 [20]BokKiKang,JinSooLee,SeKangChon,eto1.Develop— mentofself-microemulsifyingdragdeliverysystems (SMEDDS)fororalbioavailabilityenhmmememofsimvastatin inbeagledogs[J].Ira.,P^n册,2004,274:65—73. 维甲酸自乳化给药系统的体外溶出及比格犬药物动力学研究 全东琴,徐贵霞 (军事医学科学院毒物药物研究所,北京100850) 摘要:目的评价维甲酸(RA)自乳化释药系统的体外溶出及在比格犬体内的药动学.方法采用HPLC法测定比格犬 血浆药物浓度.用四条比格犬交叉实验考察RA自乳化制剂与市售胶囊体内药动学.结果实验明与市售胶囊剂相比:以 水为溶出介质,维甲酸RA自乳化制剂15min可以溶出80%以上;而市售胶囊只溶出不到5%.比格犬体内药动学研究结果 表明:与市售胶囊剂相比,自乳化制剂达峰时间提前,,,=1.25h而市售胶囊,一=2h;最大血药浓度达到市售胶囊剂的近两 倍,AUC.,)提高了67%.结论自乳化制剂可以显着提高RA的体外溶出及体内吸收. 关键词:自乳化;维甲酸;生物利用度;溶出