application of biorelevant dissolution tests

European Journal of Pharmaceutical Sciences 37 (2009) 434441

Contents lists available at ScienceDirect

European Journal of Pharmaceutical Sciences

journa l homepage: www.e lsev ier .co

Applica theperform l mpellet d

Ekarat Ja ValMaria Vea Institute of Ph 438 Fb Eurand SpA, Mc Faculty of Pha

a r t i c l

Article history:Received 28 January 2009Received in revised form 23 March 2009Accepted 27 March 2009Available online 5 April 2009

Keywords:Biorelevant diBio-DisFlow-throughIn vitroin vivoFood effectsDiclofenac sod

In vitro biorelevant dissolution tests enabling the prediction of in vivo performance of an oral modied-release (MR)dosage formweredeveloped in this study. In vitrodissolutionofMRdiclofenac sodiumpelletscontaining 100mg active ingredient was evaluated under simulated pre- and postprandial conditionsusing USP Apparatus 3 (reciprocating cylinder, Bio-Dis) and 4 (ow-through cell) and results comparedwith compendial methods using USP Apparatus 1 (basket) and 2 (paddle). In vivo, the effects of food

1. Introdu

For someber of in vivof the apprtion (IVIVCrespect to tapplied as aFDA, 1997bare productstep to drudifferent aspounds houal., 1995; DSunesen etin general (

CorresponE-mail add

0928-0987/$ doi:10.1016/j.essolution tests

cellcorrelations

ium

on the absorption of diclofenac sodium from the pellet dosage form were investigated by administeringthe product to 16 healthy volunteers pre- and postprandially in a crossover-design study. The in vitroresults were compared with the in vivo data by means of Level A in vitroin vivo correlation (IVIVC) andWeibull distribution analysis. The compendial dissolution tests were not able to predict food effects. Thebiorelevant dissolution tests predicted correctly that the release (and hence absorption) of diclofenacsodium would be slower in the fed state than in the fasted state. No signicant differences in extent ofabsorption due to changes in extent of release were predicted or observed. The results demonstrate goodcorrelations between in vitro drug release and in vivo drug absorption in both pre- and postprandial statesusing the biorelevant dissolution test methods.

2009 Elsevier B.V. All rights reserved.

ction

years attempts have been made to minimize the num-o studies necessary to approve new drug products. Oneoaches currently used is the in vitroin vivo correla-) (Uppoor, 2001; Emami, 2006; US FDA, 1997b). Withhe IVIVC concept, in vitro (mainly dissolution) tests aretool to predict drug product performance in vivo (US

; USP 30, 2008). The best candidates for IVIVC analysiss for which dissolution is the overall rate-determiningg absorption. There are many studies demonstratingpects of IVIVC for products containing BCS Class II com-sed in immediate-release (IR) dosage forms (Amidon etressman and Reppas, 2000; Wei and Lbenberg, 2006;al., 2005) and for modied-release (MR) dosage formsKortejrvi et al., 2002; Balan et al., 2001; Frick et al.,

ding author. Tel.: +49 69 798 29685; fax: +49 69 798 29694.ress: [email protected] (E. Jantratid).

1998; Takka et al., 2003; Wingstrand et al., 1990; Abrahamssonet al., 1994). Some of these studies employed dissolution mediadescribed in thepharmacopeias (so-called compendial approach)(Kortejrvi et al., 2002; Balan et al., 2001; Frick et al., 1998; Takka etal., 2003), while others added synthetic surfactants to compendialmedia (Wingstrand et al., 1990; Abrahamsson et al., 1994; Rossi etal., 2007). As these conditions do not comprehensively representthe gastrointestinal (GI) tract environment, it can be inferred thatthe results can only be interpreted on an empirical basis.

As a part of a general drive to develop predictive in vitro models,biorelevant media were proposed and have evolved over the lastdecade as a tool for in vitro biorelevant dissolution tests (Dressmanet al., 1998; Galia et al., 1998; Nicolaides et al., 1999; Vertzoni etal., 2004). Recently, the media have been updated to more nearlyrepresent both the pre- and postprandial states in the proximal gut(Dressman et al., 2007; Lue et al., 2008; Jantratid et al., 2008b). Themedia compositionshavebeenne-tunedaccording to thephaseofdigestion both in the stomach and the upper small intestine, sincethese factors can dramatically affect drug solubility and dissolutionrate. By employing these biorelevant media it should be possible to

see front matter 2009 Elsevier B.V. All rights reserved.jps.2009.03.015tion of biorelevant dissolution tests toance of diclofenac sodium from an oraosage form

ntratida,, Vincenzo De Maiob, Emanuela Rondab,rtzonic, Jennifer B. Dressmana

armaceutical Technology, Johann Wolfgang Goethe University, Max-von-Laue-Str. 9, 60ilan, Italyrmacy, National and Kapodistrian University of Athens, Athens, Greece

e i n f o a b s t r a c tm/locate /e jps

prediction of in vivoodied-release

entina Mattavelli b,

rankfurt am Main, Germany

E. Jantratid et al. / European Journal of Pharmaceutical Sciences 37 (2009) 434441 435

predict the effects of food on drug exposure in vivo, which can differby as much as one or two orders of magnitude.

One limitation of the compendial approaches to simulating thein vivo relearesults areited numbethe compenmediumduto the changally exposeddrug produtory of thepolymer cosubsequentreduction inupon passaset up the isuch away (effects can

The USP(ow-throufrom the dolating the GBio-Dis andtion mediahistory ofgenerate an

Diclofenwas used alated intoactive pharammonio mas a releasfrom MR pow-througassembly).hence its abulating theet al., 2005et al., 2008netic data oA IVIVC (U1965) and R2005) wermodel.

2. Materia

2.1. Materia

Diclofentured by Eugrade and ophosphatidwas purchaerylmonoolot 4010380Denmark. HMallinckroddrous, lot 1Eur., 0.51UAG, Buchs, Spurchasedmany. Sodi

Riedel-deHan, Seelze,Germany. Sodiumtaurocholate (NaTC, >99%pure, lot 2006040099) was purchased from Prodotti Chimici e Ali-mentari SpA, Basaluzzo, Italy. Ammonium dihydrogen phosphate,

omethane, sodiumdihydrogenphosphatemonohydrate anddrofuran (THF)were obtained fromMerckKGaA,Darmstadt,ny. Iilanchlohyd

arloaseLab

anti

amed uhe sya W

e Arrzu, S4, 5ed ocetos sete inje75namo

by en) pTheC. Tatermcon

zu Sectrotationm,rch Perck0.4%.0mn vo.

odie

lofennd t

led innac spe Ar, w

ssolu

lity cand

is IIIrougsolutformse proles of dosage forms with MR properties is thattypically run in one medium at a time. Only in a lim-r of cases (most notably enteric coated products) doesdial method call for a change in the composition of thering the test (USP 30, 2008). This is obviously in contrasting environment to which the dosage forms are gener-as they pass through the GI tract. Additionally, for MR

cts, the release of drug may be inuenced by the his-dosage form after ingestion. For example, swelling of

mponents under gastric conditions may inuence therelease under intestinal conditions or lead to such amechanical resistance that the dosage form breaks up

ge through the ileocecal valve. Thus, it is desirable ton vitro release test conditions for MR dosage forms ini.e. with a series ofmedia in one experiment) that thesebe observed and predicted.Apparatus 3 (reciprocating cylinder, Bio-Dis) and 4

gh cell) offer the advantages of determining releasesage form under various, consecutive conditions simu-I physiology. The release experiments performed withow-through cell can be set up with a series of dissolu-in one single run, thus making it possible to mimic thethe dosage form as it passes through the GI tract and toIVIVC on an a priori basis.ac sodium, a non-steroidal anti-inammatory drug,s a model compound in this study. It was formu-

capsules containing MR pellets consisting of 100mgmaceutical ingredient (API) per dosage unit usingethacrylate copolymer type A (NF) (USP 30, 2008)

e-modifying agent. The release of diclofenac sodiumellet dosage forms was compared using Bio-Dis andh cell with USP Apparatus 1 (basket) and 2 (paddleFood effects on the release of diclofenac sodium, andsorption, were predicted using biorelevant media sim-fasted and fed states in the human GI tract (Vertzoni; Fotaki et al., 2005; Dressman et al., 2007; Jantratidb). In vitro results were compared with pharmacoki-btained in the fed and fasted human subjects. LevelSP 30, 2008; Levy and Hollister, 1964; Levy et al.,RSBW (Weibull) distribution analysis (Langenbucher,

e applied to assess predictability of the in vitro

ls and methods

ls

ac sodium capsules (lot P200750226) were manufac-rand SpA. Acetonitrile and methanol were of gradientbtained from Merck KGaA, Darmstadt, Germany. Eggylcholine (Lipoid E PC, 97.9% pure, lot 108027-1/045)sed from Lipoid GmbH, Ludwigshafen, Germany. Glyc-leate (GMO, RyloMG19Pharma, 99.5%monoglyceride,689) was a gift from Danisco Specialities, Brabrand,ydrochloric acid (37%, fuming) was purchased fromt Baker, Milan, Italy. Calcium chloride (97% pure, anhy-309867), ortho-phosphoric acid (85%) and pepsin (Ph./mg, lot 1296935) were obtained from Fluka Chemiewitzerland. Maleic acid (99% pure, lot 056K5473) wasfrom SigmaAldrich Chemie GmbH, Steinheim, Ger-um oleate (82.7% pure, lot 61720) was obtained from

dichlortetrahyGermaBDH, Msodiumsodiumfrom CUSP lipProtein

2.2. Qu

Theanalyztem. Tpump,todiodShimad125mmcolumnconsist29.5% aratewaple. Thset at 2

Thetiedsolutio(95:5).at 50

trile/wsystemShimatning spWorksumn, 5UpchuI.D. (M53% ofset at 1injectio275nm

2.3. M

Dic2mm aand ldiclofemer typolyme

2.4. Di

Quapaddle(Bio-Dow-ththe disdosagesopropyl alcohol and n-hexane were purchased from, Italy. Glacial acetic acid, sodium acetate trihydrate,ride, sodium dihydrogen phosphate monohydrate androxidepelletswereall of analytical gradeandpurchasedErba Reagenti SpA, Milan, Italy. Pancreatin powder (72Units/mg, lot 1279-0057) was obtained from Scienticoratories LLC, WI, USA.

tative analysis of diclofenac sodium

ounts of diclofenac dissolved in the samples weresing a validated isocratic reversed phase HPLC sys-stem consisted of a Waters Alliance 2695 quaternaryaters Alliance 2695 autosampler, a Waters 996 Pho-ay Detector (Milford, MA, USA), an integrator (CR5Ahimadzu, Kyoto, Japan) and an RP-18-e column, 5m,mm LiChrospher 100 connected with a RP-18-e guard-m, 4mm4mm LiChrospher 100. The mobile phasef 63% of 0.05M ammonium phosphate bufferpH 5.0,nitrile and 7.5% tetrahydrofuran (by volume). The owat 1.2mL/min resulting in a run time of 15min per sam-ction volumewas 20L. The detectionwavelengthwasm.unts of diclofenac in the plasma samples were quan-xtracting the acidied (with 0.5N hydrochloric acidlasma with an n-hexane/isopropyl alcohol mixtureorganic layer was evaporated under nitrogen streamhe dried residues were reconstituted with acetoni-

ixture (1:1) and then injected into theHPLC. TheHPLCsisted of a Spectra Physics SP 8800 ternary pump, aIL-9A autosampler, a Spetra Physics SP 8480 XR scan-photometer, a PCAX2 (Epson)withOmega 2Analyticalsoftware integrator (Perkin Elmer) and an HS 5C8 col-

15mm4.6mm I.D. (Perkin Elmer) connected with anerisorb RP-18 guardcolumn, 3040m, 3 cm2.1mm). The mobile phase consisted of 47% acetonitrile andortho-phosphoric acid (by volume). The ow rate was

L/min resulting in a run time of 10min per sample. Thelume was 50L. The detection wavelength was set at

d-release formulations

ac sodium pellets (with a pellet size of approximatelyhe average API content of 100.7%) were manufacturedhard gelatin capsules at a label strength of 100mg of

odiumper dosage unit. Ammoniomethacrylate copoly-(NF) (USP 30, 2008), a pH-independent, permeable

as used as a release-modifying agent.

tion testing

ontrol (QC) compendial methods using the basket andbiorelevant methods using the reciprocating cylinderExtended Release Tester, Varian Inc., CA, USA) and theh cell (Sotax S.r.l., Bergamo, Italy)were used to evaluateion behavior of diclofenac sodium from the MR pellet.

436 E. Jantratid et al. / European Journal of Pharmaceutical Sciences 37 (2009) 434441

Table 1Biorelevant dissolution media simulating the preprandial conditions in the gastrointestinal tract.

Medium FaSSGFa New-FaSSIFb Half-FaSSIF FaSSIF-sans SCoFc

Bile secretions- NaTC 1.5mM - -- Lecithin 0.2mM - -

Enzyme- Pepsin - - -- Pancreatin - - -

pH 7.0 7.5 5.8Osmolality (m 270 270 295Buffer capacity 10 10 29

Abbreviations tinal Fluid; SCoFSimulated Colonic Fluid; NaTCsodium taurocholate.a Vertzoni eb Dressmanc Fotaki et a

Table 2Biorelevant di .

Medium Half-FeSSIF FaSSIF-sansc SCoFd

Bile secretions- NaTC- Lecithin

Lipolytic prod- GMO- Sodium ole

Enzyme- Pepsin- Pancreatin

pHOsmolality (mBuffer capacity

Abbreviationsa Jantratid eb Dressmanc The condit

conditions.d Fotaki et a

2.4.1. QualiThe QC

900mL phoket rotationThe paddleket methodat 75 or 12The sampliwasperformume withdpoint. The sroethyleneat 275nm.

2.4.2. Biore2.4.2.1. Bio-the Bio-Dissodium fropostprandiadissolutionexperiment(Klein et alpreparation2005; Fotak2008b). Theof 220mL p80M 3.0mM20M 0.2mM

0.1mg/mL -(lipase Unit/mL) - -

1.6 6.5Osm/kg) 120.7 180(mEqL1 pH1) - 10

: FaSSGFFasted State Simulated Gastric Fluid; FaSSIFFasted State Simulated Intest al. (2005).et al. (2007) and Jantratid et al. (2008a,b).l. (2005).

ssolution media simulating the postprandial conditions in the gastrointestinal tract

FeSSGFa New-FeSSIFb

- 7.5mM- 2.0mM

ucts- 5.0

ate - 0.8

- -(lipase Unit/mL) - 1005.0 5.8Osm/kg) 400 390(mEqL1 pH1) 25 25

: FeSSGFFed State Simulated Gastric Fluid; FeSSIFFed State Simulated Intestinal Fluid;t al. (2008a,b). This medium contains 50% full fat (3.5%) UHT-milk.et al. (2007).ions during the late postprandial state in the upper small intestine are similar to the pr

l. (2005).

ty control methodtest conditions for the basket method consisted ofsphate buffer, pH 6.8 as a dissolution medium, the bas-speed of 50 rpm, and a temperature of 37 C0.5 C.

method employed the same conditions as for the bas-, except for the paddle rotational speed, which was set5 rpm. Experiments were conducted in six replicates.ng times were 30, 60, 120, 180 and 240min. Samplingedautomatically through the samplingdevice. Thevol-

rawn was approximately 5mL for each sampling timeamples were ltered through a 0.45m polytetrauo-(PTFE) lter and then analyzed spectrophotometrically

levant methodsDis method. A biorelevant, pH-gradient method usingtester was applied to simulate release of diclofenac

m the MR pellet dosage form in the GI tract pre- andlly. The biorelevant media used in this study and thetest set-up are shown in Tables 13. The dissolutional design was modied from that proposed previously., 2005, 2008). The detailed compositions and mediahave mostly been described elsewhere (Vertzoni et al.,i et al., 2005; Dressman et al., 2007; Jantratid et al.,dissolution conditions consisted of a media volume

er vessel with a dip rate of 10dpm. The top and bot-

tom mesh spreprandial540 and 750270, 330, 4was maintareplicates. Sdevice (Van

Table 3Preprandial antests of MR dic

Segment of th

StomachDuodenum/JejJejunum/IleumDistal ileumColon

Segment of th

StomachDuodenum/JejJejunum/IleumDistal ileumColon3.0mM - -1.0mM - -

- - -- - -

- - -- - -6.5 7.5 5.8270 270 29510 10 29

GMOglyceryl monooleate

eprandial state. Therefore, FaSSIF-sans was used for simulating both

ize was 405m (40 mesh). The sampling times for thesimulation were 30, 60, 105, 150, 210, 270, 360, 450,min and for the postprandial simulation 120, 165, 210,

50, 630 and 810min. The temperature in the vesselsined at 370.5 C. Experiments were conducted in sixamplingwasperformedautomaticallyusinga samplingKel VK 8000 Dissolution Sampling Station, Varian Inc.,

d postprandial pH-gradient schedule for the biorelevant dissolutionlofenac sodium pellets using Bio-Dis method.

e GI tract pH-gradient preprandial Residence time (min)

Biorelevant medium pH

FaSSGF 1.6 60unum New-FaSSIF 6.5 45

Half-FaSSIF 7.0 45FaSSIF-sans 7.5 120SCoF 5.8 480

e GI tract pH-gradient postprandial Residence time (min)

Biorelevant medium pH

FeSSGF 5.0 120unum New-FeSSIF 5.8 45

Half-FeSSIF 6.5 45FaSSIF-sans 7.5 120SCoF 5.8 480


CA, USA). The sample volume withdrawn was approximately 5mL.With the exception of FeSSGF, the samples were ltered througha 0.45m PTFE lter and then analyzed by HPLC. The amount ofdrug releasthe innityof vessels coset-up in thrstve rowwas exhausin all vessel(100mg), th

2.4.2.2. Flowtester equipwas used tpellets duriA 5mm-sizof 1.7 g of 1glass bead,diameter, Mcell. DuringExperimenttion mediaand ascendfor the Bioous fasted aow rates wow-cell waexchanged

The conlyzed by HPexperimentltration. Inferred to a tfor 1min, thclear supern

2.5. Compa

The MRvolunteersold (range5283kg); mand fed staperiod of ondard proceddrug absorpDeclarationlocal ethicssamples wepleswere cowere determ

2.6. Analysi

Differenusing the mfactor (f2) a

f2 = 50 log

where n isof the refertest at time

in the percent (%) drug dissolution between the two curves. Valuesof 50 or above (50100) ensure similarity (difference10%) of thecurves.

alysi

-comacokrsionA) annd,Wet thtratincenwhilrisons.

vitro

plasd p

bsorbe rencen

istrateers (ere taFa v

ll dis

max

Wt isaxi

n parimerocee Siing Senervantta, i.e pare co965rison

ults

relev

elevan

simu, i.e.wasvantof mreced pons, as andthe

ity aned in the FeSSGF was indirectly determined by usingpoint approach (Klein, 2005) by adding one more rowntaining phosphate buffer, pH6.8 to the series ofmediae postprandial state. After the release was tested in thes, pelletswereexposed to thismediumuntil the release

ted (20dpm, 6h). By subtracting the cumulative releases analyzed from the label strength of the dosage forme drug release in the rst row can be estimated.

-through cell method. The ow-through dissolutionpedwith22.6mmdiameter test cells andapistonpumpo evaluate the release of diclofenac sodium from theng exposure to the biorelevant, pH-gradient methods.ed glass bead was placed in the tip of the cell. A totalmm-sized glass beads were added above the 5mmwhile a glass ber (MNGF1, 0.7m pore size, 25mmachery-Nagel, Germany) was placed on the top of thethe experiment, the capsule was mounted on a holder.s were performed in triplicate at 370.5 C in dissolu-simulating the compositions of gastric, small intestinaling colonic contents in the fasted and fed states (as-Dis experiments). Duration of exposure to the vari-nd fed state simulating media and the correspondingere adapted from Fotaki et al. (2005). Fluid exiting thes collected in a volumetric cylinder. The cylinders were

every 20min up to 420min.tent of diclofenac sodium in each cylinder was ana-LC. Except for FeSSGF, the samples collected from theswere immediately injected into the HPLC system aftercaseof FeSSGF, 1mLof the collected sampleswas trans-ube and 2mL of acetonitrile was added. After vortexinge sample was centrifuged for 10min at 4000 rpm. Theatant was then injected into the HPLC system.

rative bioavailability studies

diclofenac sodium pellets were given to 16 healthy[nine males and seven females, mean age24.9 years2033 years old); mean body weight65kg (rangeean height173 cm (range 164185 cm)] in the fasted

tes on a randomized crossover basis with a wash-oute week. The study was conducted following the stan-ure of the US FDA for the assessment of food effects ontion (US FDA, 2002) and the recommendations of theof Helsinki. The study protocol was approved by thecommittee prior to the beginning of the study. Bloodre collected every hour up to 12h and the last sam-llected at 24h. Thediclofenac concentrations in plasmained using the aforementioned HPLC method.

s of in vitro dissolution data

ces in the in vitro dissolution proles were assessedodel-independent approach based on the similarity

s follows (US FDA, 1997a):{[1 + 1

nnt=1(Rt Tt)2

]0.5 100

}(1)

the number of time points, Rt is the dissolution valueence at time t, and Tt is the dissolution value of thet. The f2 is basically a measurement of the similarity

2.7. An

Nonpharmtion veCA, USRedmointerprconcendrug costates,compaanalyse

2.8. In

Thepre- andrug aimpulsdrug coadminvoluntles w

TheWeibu

Wt = Wwhereis the mlocatiois the tof the pwith thtion us

Togbioreletion dathe timtion) aet al., 1compa

3. Res

3.1. Bio

Bior(Dressmbettercessorsstudybioreledesignon thepre- anpositiochangetract insolubils of in vivo pharmacokinetic data

partmental analysis was applied to the evaluation ofinetic parameters using WinNonlin Professional Edi-4.1 software (Pharsight Corporation, Mountain View,

d Microsoft Ofce Excel 2003 (Microsoft Corporation,A,USA). Students paired t-testwasused to statistically

e differences between the area under the plasma drugontime curves (AUC0t) and the maximum plasmatration (Cmax) after administration in the fed and fastede Wilcoxons signed-rank test was used for the Tmax. A probability level of 0.01 was applied to all statistical

in vivo correlations

ma drug concentrationtime proles obtained in theostprandial states were deconvoluted to the fractioned (Fa) proles using WinNonlin software. The unit

sponse was determined from the literature plasmatrationtime data obtained following the intravenousion of 50mg diclofenac sodium to seven healthy femaleWillis et al., 1979). The fraction drug dissolved (Fd) pro-ken directly from the dissolution data.s Fd curve comparisons were determined using thetribution as described by the following equation:

[1 e[(t)/d] ] (2)the fraction of drug dissolved/absorbed at time t,Wmaxmum cumulative fraction dissolved/absorbed, is theameter (the lag time before the onset of dissolution), dparameter (provides information about the overall ratess), and is the shape parameter. The proles were tgmoidalWeibull distribution with a slight modica-igmaPlot software, version 10.0 (Erkrath, Germany).ate LevelA IVIVC thedissolution results fromtheQCandmethods were plotted against the in vivo drug absorp-e. the relationship between Fa and Fd values, in whichirs in vitro (drug dissolution) and in vivo (drug absorp-rrelated (USP 30, 2008; Levy and Hollister, 1964; Levy). No time scale corrections were made for the prole.

and discussion

ant dissolution media

ant dissolution media have been updated recentlyet al., 2007; Lue et al., 2008; Jantratid et al., 2008b) tolate theproximalhumanGI conditions than their prede-FaSSIF and FeSSIF (Galia et al., 1998). In fact, the presentperformed during the development of the updateddissolution media (Jantratid et al., 2008b) and theedia compositions described herein was based mostlynt in vivo data collected from human aspirates in thestprandial states (Kalantzi et al., 2006). Themedia com-s tabulated in Tables 1 and 2, reect the physiologicalthe on-going digestive processes along the human GI

fasted and fed states. Crucial parameters affecting drugd dissolution including (i) the levels of bile secretions,


(ii) the presence of lipolytic products and enzymes, and (iii) the pH,buffer capacity and osmolality, were all taken into consideration. Inthis study, media representing the fasted and fed stomach (FaSSGFand FeSSGFFaSSIF, andFaSSIF-sansadopted froDressman ewas used folower partponents is vused as a dicolon in bot

3.2. Quanti

Validatiosodium assation (LOD)w0.3g/mL.concentratiof the assay

For the athe LOD waery of 99.7102000ngthan 10%.

3.3. Dissolu

3.3.1. QualiFig. 1A s

the MR pellgave compamethod, rotdifferences(f2 value =6all test cond

3.3.2. BioreThe diss

lets using tFig. 1B. In tto simulatethe pre- anmedia comin the differthe dosageDependingmultiparticcally differeet al., 1986;thus arrivalis generallythe gastricthan monol1991). Baselets in diffeconditions (were appliedescribed in

The resuthe releasethan in anTable 4) ind

issolution prole comparison of cumulative diclofenac sodium dissolu-ime proles using (A) the QC methods (basket and paddle); and (B) theant methods in the fasted and fed state using the Bio-Dis and ow-throughs.

teristics in the fasted and fed states using either Bio-Dis-through cell. Owing to the weakly acidic properties ofnac (pKa 3.8) (OConnor and Corrigan, 2001), the release inted stomach in normal acid secretors is expected to be poor.er, the gastric residence time is short and it can be expectede release would go up immediately after the dosage formen emptied into the duodenum. By contrast, after ingestionmeal, the gastric pH initially increases (Russell et al., 1993;

parameters derived from the fraction drug absorbed and the fraction drugd proles.

Weibullparameters

Fain vivoa FdBio-Disb FdFlow-throughc

d (min) 243.7 167.3 193.2 1.7 2.8 2.9

d (min) 420.5 249 289 2.6 4.3 2.7

n vivofraction of drug absorbed deconvoluted from the in vivo plasma

io-Disfraction of drug dissolved obtained from Bio-Dis method.low-throughfraction of drug dissolved obtained from ow-through cell), the small intestine in the fasted (New-FaSSIF, Half-FaSSIF-sans) and fed state (New-FeSSIF, Half-FeSSIF, and), and the ascending colon (SCoF) were developed ormthe literature (Vertzoni et al., 2005; Fotaki et al., 2005;t al., 2007; Jantratid et al., 2008b). FaSSIF-sans (pH 7.5)r both the pre- and postprandial states to simulate theof the small intestine, where reuptake of the bile com-irtually complete and pH is higher. Likewise, SCoF was

ssolutionmedium to represent the conditions in humanh the pre- and postprandial states (Fotaki et al., 2005).

tative analysis of diclofenac sodium

n of the chromatographic method for diclofenacy from the in vitro tests showed that the limit of detec-as0.09g/mLand the limit of quantication (LOQ)was

The mean recovery of 99.0% was observed within theon range of 15150g/mL. The accuracy and presicionwere less than 5%.nalysis of diclofenac sodium concentration in plasma,s 2ng/mL and the LOQ was 10ng/mL. The mean recov-% was observed within the concentration range of/mL. The accuracy and presicion of the assay were less

tion testing

ty control methodshows the dissolution proles of diclofenac sodium fromets using the QC methods. Basket and paddle methodsrable dissolution results. Additionally, using the paddleational speeds of 75 and 125 rpm showed no signicantin the release rate of diclofenac sodium form the pellets7.3). Approximately 85% drug release was observed initions within 120min.

levant methodsolution proles of diclofenac sodium from the MR pel-he Bio-Dis and ow-through methods are shown inhis study, the biorelevant dissolution tests were usednot only differences in GI uid compositions betweend postprandial states, as reected by the dissolutionpositions, but also differences in the residence timesent parts of the GI tract, as reected by the passage ofform in the biorelevant dissolution test set-up (Table 3).on the dimension of the dosage units, e.g. monolithic vsulate dosage forms, the residence times can be drasti-nt especially in the stomach (Hardy et al., 1993; DavisCoupe et al., 1991). For example, gastric emptying andat regions in the small intestine with higher pH valuesquicker in the fasted state than in the fed state. Further,residence time is usually shorter for multiparticulatesiths (Hardy et al., 1993; Davis et al., 1986; Coupe et al.,d on the literature data, the residence times of the pel-rent regions of the human GI tract in the fasted and fedHardy et al., 1993; Davis et al., 1986; Coupe et al., 1991)d to the design of the biorelevant dissolution tests, asTable 3.

lts in Fig. 1B show that under postprandial conditionsof diclofenac sodium from the pellets would be slowerempty stomach. Weibull analysis (d and values;icates an apparent difference between the dissolution

Fig. 1. Dtion vs tbiorelevapparatu

characor owdiclofethe fasHowevthat thhas beof the

Table 4Weibulldissolve

Fasted

Fed

a Faiproles.

b FdBc FdF

method.


Fig. 2. (A) Plasing a single o16 healthy volconcentration50mg diclofenthe mean plas

Dressman efed gastric2008b). Atconditions iHowever, thfaster in thetying inducwould reachcan occur, a1986; Coupas the pH inlution resuldrug in both

3.4. Compa

Fig. 2A ssignicant dles followiin the pre-nicantly d

ean flutionsted st

-rank, wasFig. 3. Mdeconvoin the fa

signedas AUCma diclofenac sodium concentrationtime proles obtained follow-ral administration of modied-release diclofenac sodium pellets tounteers in the fasted and fed states; (B) plasma diclofenac sodiumtime proles obtained following an intravenous administration ofac sodium to 7 volunteers (Willis et al., 1979). Each point representsma diclofenac sodium concentration.

t al., 1990; Kalantzi et al., 2006), as reected by thedissolution medium, FeSSGF (pH 5.0) (Jantratid et al.,this pH, diclofenac is more soluble than in the acidicn the fasted state and part of the drug can be released.e results show that the dissolution rate is not actuallyfed gastric conditions. With the delayed gastric emp-

ed by food commonly observed in vivo, the dosage formthe proximal small intestine, where signicant release

fter approximately 24h (Hardy et al., 1993; Davis et al.,e et al., 1991). Subsequently, along the small intestine,creases the release is expected to continue. The disso-ts indicate that after approximately 6h the release offed and fasted states are expected to be complete.

rative bioavailability studies

hows a slow onset of absorption after meal intake, withifferences in the plasma drug concentrationtime pro-ng oral administration of MR diclofenac sodium pelletsand postprandial states. Both Cmax and Tmax were sig-ifferent (p0.01, Students paired t-sults are in agreement with previously reported data., 1981; Riad et al., 1995).

nalysis and curve comparisons

volution of pharmacokinetic dataintravenous data was available from this study, theytaken from the literature (Fig. 2B) (Willis et al.,

rform numerical deconvolution using the WinNonlin

herefore, the deconvoluted data cannot be inter-00% accurate. By tting the intravenous plasma drugontime data to the Exponential Decay regressionPlot program, tri-exponential functionswere found toe prole appropriately. The unit impulse response wasted from the coefcients of tri-exponential equation.ows the Fa proles obtained following the numericalion of the plasma proles in the fasted and fed states.erns to those measured in the in vitro dissolution pro-ed from the biorelevant methods were observed. Bye QC dissolution methods give dissolution patterns thatly different from the in vivo absorption proles.comparisonsbull distributionhaspreviouslybeenapplied to theeval-IVC (Nicolaides et al., 2001; Jantratid et al., 2008a). Inwas used to explain the incremental changes in the Fa

es over the same time-frame. As demonstrated in Fig. 4,les from both the QC and the biorelevant methods arethe Fa proles in the fastedand fed states.However, the

om the QC dissolution methods could be excluded fromderations since it runsmuch ahead of the biorelevant Fdroles, and is exponential in shape. A sigmoidal shapee well described by using the Weibull distribution wasr the Fd proles from the Bio-Dis and the ow-throughs, as well as the Fa proles. The parameters obtainedll analysis of these proles are given in Table 4. The dch represent the overall rate of dissolution/absorption,t the time-frames required for thedissolutioncurves areter than those required for the absorption curve bothand fed states. The values, which describe the shape

es, indicate that although the curve increments in vitro


Fig. 4. Compadissolution tes(B). The data w

and in vivothe fed stat

The obsevivo absorpvariations ismall intestup in this stin vivo dataCoupe et al.gastric residlead to in vprole. Thetion to theof diclofenachanges, e.gproperties aTherefore thstate, can btine. These,affected by

3.5.3. In vitFig. 5dem

states, the l(r2 >0.95), i

n vitrothe tdisso

methsati

leaserepr

al (thFig. 5. Ibetweenand drug

lutionsodiumvitro rein the pprandirison of the fraction drug dissolved (Fd) obtained from the biorelevantts vs the fractiondrugabsorbed (Fa) in the fasted state (A) and fed stateere tted to Weibull distribution.

are both sigmoidal ( >1) the changes (particularly ine) are more obvious in vitro than in vivo.rved difference between the in vitro dissolution and intion, especially in the fed state, can be explained by then the gastric residence time and the variations of theinal pHs. The passage time used for the dissolution set-udywas set as2h for the fedstate stomach.However, therange from2 to 4h (Hardy et al., 1993; Davis et al., 1986;, 1991). It can therefore be expected that the longer fedence time invoked for the biorelevant methods would

itro dissolution proles closer to the in vivo absorptionother reason for the deviation of the in vitro dissolu-

in vivo absorption is that the solubility and dissolutionc sodium is known to be inuenced strongly by pH. conditions in the GI tract, owing to the weakly acidicnd the pKa of the drug itself (Chuasuwan et al., 2009).e releaseof drug in vivo, particularly in thepostprandiale very sensitive to the conditions in the small intes-in turn, show interindividual variation as well as beingthe type of meal administered (Clarysse et al., 2009).

roin vivo correlationsonstrates the LevelA IVIVCplots. Inboth fastedand fed

east square regressions yield essentially linear patternsndicating that the proposed in vitro biorelevant disso-

cell, the in vpre- and po1.2928, resp

4. Conclus

The biorthis study wperformancdrug releascompendiathe time paresults indierally apprMR diclofenslightly sup

Acknowled

Part of tMeeting antion Center,

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ions

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Application of biorelevant dissolution tests to the prediction of in vivo performance of diclofenac sodium from an oral modified-release pellet dosage formIntroductionMaterials and methodsMaterialsQuantitative analysis of diclofenac sodiumModified-release formulationsDissolution testingQuality control methodBiorelevant methodsBio-Dis methodFlow-through cell method

Comparative bioavailability studiesAnalysis of in vitro dissolution dataAnalysis of in vivo pharmacokinetic dataIn vitro-in vivo correlations

Results and discussionBiorelevant dissolution mediaQuantitative analysis of diclofenac sodiumDissolution testingQuality control methodsBiorelevant methods

Comparative bioavailability studiesIVIVC analysis and curve comparisonsDeconvolution of pharmacokinetic dataCurve comparisonsIn vitro-in vivo correlations

ConclusionsAcknowledgementReferences

application of biorelevant dissolution tests

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