International Journal of CURRENT PHARMACEUTICAL RESEARCH

ISSN- 0975-1491 Vol 2, Issue 4, 2010

ResearchArticle

BIOEQUIVALENCEANDPHARMACOKINETICEVALUATIONOFTWOFORMULATIONSOF PARACETAMOLER650MG:ASINGLEDOSERANDOMIZEDTWOPERIODCROSSOVER COMPARISONINHEALTHYINDIANADULTVOLUNTEERS

DHANESHWARSHEP1,ASHWINIOJHA2,SWETAPATEL3,RAJESHWARIRATHOD4,MANISHNIVSARKAR5,VIJAYA JAISWAL6ANDHARISHPADH7
1Sr.Manager,MedicalServices,TroikaaPharmaceuticalsLtd.,Ahmedabad,2ScientistA,DepartmentofBioanalytical,B.V.PatelPERD Centre,Ahmedabad,3ResearchAssistant,DepartmentofBiostatistics,B.V.PatelPERDCentre,Ahmedabad,4ScientistA,Departmentof Bioanalytical,B.V.PatelPERDCentre,Ahmedabad,5JointDirector,DepartmentofPharmacologyandToxicology,B.V.PatelPERDCentre, Ahmedabad,6VicePresident,MedicalServices,TroikaaPharmaceuticalsLtd,CommerceHouse1,SatyaMarg,Bodakdev,Ahmedabad 54,PhoneNo91792685624245,Fax:917926856246.7Director,B.V.PatelPERDCentre&ProjectDirectorNIPER,Ahmedabad.

Email:medicalservices@troikaapharma.com Received:10Jun2010,RevisedandAccepted:11July2010 ABSTRACT Background:Paracetamolisoneofthemostpopularandwidelyuseddrugsforthetreatmentofpainandfever.Duetoitsgoodtolerabilityprofile, paracetamolisoftentheanalgesicorantipyreticofchoice,especiallyinpatientsinwhomsalicylatesorothernonsteroidalantiinflammatorydrugs arecontraindicated.OBJECTIVE:Theaimofthisstudywastocomparethebioavailabilityandtolerabilityof2oralformulationsofparacetamolER 650mg.Methods:Thissingledose,randomized,singlelabel,2periodcrossoverstudyinhealthyIndianadultvolunteerswasconductedatPERD Centre,Ahmedabad.SubjectsreceivedparacetamolER650mgofeithertestorreferenceformulationwithawashoutperiodof7days.Afterstudy drugadministration,serial bloodsamples were collected over aperiod of24 hours.Plasmawasanalyzed forparacetamol concentration using a validated highperformance liquid chromatography method. Pharmacokinetic (PK) parameters Cmax, Tmax, t1/2, AUC0t, AUC 0, and kel, were determinedforthe2paracetamolformulations.TheformulationsweretobeconsideredbioequivalentifthelogtransformedratiosofCmax,AUC0t, andAUC0werewithinthepredeterminedbioequivalencerangeof80%to125%.Results:Atotalof18subjectswereenrolled(meanBMI21.97,a meanageof29.4years,meanweightof60.7kgandameanheightof166.6cm).Nosignificantdifferenceswerefoundbasedonanalysisofvariance, withmeanvaluesand90%confidenceintervalsoftest/referenceratiosfortheseparametersasfollows:Cmax,6.02versus6.17g/mL(89.9109.9); AUC0t,30.54versus31.12g.hr/mL(92.31104.1);andAUC 0,31.36versus31.69g.hr/mL(91.58106.6).CONCLUSION:InthesehealthyIndian volunteers,resultsfromthePKanalysissuggestedthatthetestandreferenceformulationsofparacetamolER650mgtabletswerebioequivalent, basedontheregulatorydefinition. Keywords:Paracetamol,Bioequivalenceevaluation INTRODUCTION Paracetamol is one of the most popular and widely used drugs for thetreatmentofpainandfever.1Duetoitsgoodtolerabilityprofile, paracetamolisoftentheanalgesicorantipyreticofchoice,especially in patients in whom salicylates or other nonsteroidal anti inflammatorydrugsarecontraindicated.2 Paracetamoliswellabsorbedfromtheproximalsmallboweland is notsubjecttosignificantfirstpassmetabolismintheliver,withoral bioavailability estimated at between 6389% in adults. 3,4 Paracetamolisnotsignificantlyboundtoplasmaproteins,andhasa volume of distribution of 0.71 L/kg. Maximal analgesic and antipyreticactivityoccurs12hafterpeakplasmalevels, 3,5andthe time to achieve this varies with the route of administration. Peak plasma concentration (Cmax) is achieved approximately at 45 min.3 Metabolism of paracetamol occurs primarily in the liver, while elimination occurs almost entirely through the kidney. Following absorptionoftherapeuticdoses,approximately90%ismetabolised by glucuronidation and sulphation to form nontoxic metabolites, whichareexcretedintheurine. Paracetamol,likemanyotheranalgesics,hasashorthalflifearound 23 hours which necessitates frequent dosing.6 In UK the recommendedregimenis5001000mgevery46hours.However, it would be advantageous if the duration of action were longer so that fewer daily doses could maintain therapeutic plasma levels. This wouldimprovepatient convenienceand complianceand beof benefittothepatientatnighttime.6 A generic version of the extended release formulation of paracetamol has been developed by Troikaa Pharmaceuticals Ltd, Indiawhichcombinesextendedandimmediatereleaseparacetamol in a bilayer tablet. This formulation (denoted as ER paracetamol) hasbeendesignedtobetakenthreetimesdaily.Eachbilayertablet contains 650 mg paracetamol (325 as immediate release and 325 mgasextendedrelease). Theaimofthisstudywastocomparethebioavailabilityofthenewly developedbilayertabletformulationofparacetamolwithinnovator brand Tylenol extended release caplets in healthy Indian adult malevolunteers. MATERIALANDMETHODS The study was carried out at the B. V. Patel Pharmaceutical Education and Research Development centre, Ahmedabad. All the subjects provided written informed consent to participate in the study prior to enrolment and were free to withdraw at any time duringthestudy.Thestudywasapprovedbytheinstitutionalethics committee and was conducted in accordance with good clinical practiceandthedeclarationofHelsinki. Studysubjects The study population consisted of 18, adult, male healthy subjects withmeanBMI21.97,ameanageof29.4years,meanweightof60.7 kgandameanheightof166.6cm. Design The study was designed as Double blind, Balanced, Randomized, TwoTreatment,TwoSequence,TwoPeriod,SingleDose,Crossover Bioequivalencestudywith7dayswashoutperiod. Thevolunteerswereadministeredoneofthetwostudydrugsafter anovernightfast.Thedoseadministrationwasperformedasperthe randomization schedule generated at B.V. Patel PERD Centre, Ahmedabad.Subjectsreceivedsingleoraldoseofthetestformulation (paracetamol ER 650 mg, Troikaa Pharmaceuticals Ltd. India) and referenceformulation(paracetamolER650mg,McNeil,USA).

Dhaneshwaretal. IntJCurrPharmRes,Vol2,Issue4,2831 Bloodsampling performance liquid chromatography method with ultravoilet detectionat242nm.100Lplasmasamplewasextractedintoethyl Atotalof19bloodsampleswerecollectedduringeachperiod.Blood acetateafteradditionofinternalstandard(4aminoacetophenone). sampleswerecollectedthroughanindwellingcannulaplacedinthe The organic layer was evaporated under nitrogen and the sample forearm vein using disposable syringe or with disposable syringes reconstitutedinmobilephasebeforeinjectiononhighperformance and needles. 4 mL of blood samples (including 0.2 mL discarded liquidchromatograph(Jasco,Japan).Thecolumnusedforseparation heparinisedblood)werewithdrawnatpredoseand0.25,0.50,0.75, from endogenous plasma components was HQ Sil HS C18, 250 X 1.0,1.25, 1.50,2.0,2.50,3.0, 3.50, 4.0, 4.50, 5.0, 6.0,8.0, 10.0, 14.0, 4.6mm,5mfromKyaTechnologiesandthemobilephasewas20% 24.0 hrs following drug administration in each period. After acetonitrile and 80% 10mM ammonium acetate. Precision and centrifugation,plasmaseparatedfrombloodsampleswasstored at accuracy were validated over the concentration range of 0.05 to 205Cforinterimstorageandthenat705Cuntilanalysis. 10g/mL.Theintraandinterdayprecision(%cv)ofthemethod at low, medium and high concentrations were less than 15%. The Methodofanalysis method accuracy ranged from 91 to 112%.The lower limit of The analytical method used for determination of drug quantitationwas0.05g/mLandthemethodrecoveryrangedfrom concentrations in invivo plasma samples was a validated high 72to81%. Table 1:Summary ofpharmacokineticparameters ofParacetamol,following administration ofthereferenceandtest formulations Products Parameters MEAN SD SEM %CV Cmax (g/mL) 6.17 1.47 0.35 23.76 Tmax (h) 1.06 0.72 0.17 67.75 Reference AUC0t (g.h/mL) 31.12 8.02 1.89 25.76 AUC0 (g.h/mL) 31.69 8.12 1.91 25.63 t1/2 (h) 7.00 2.10 0.50 30.02 Kel (h )
1

Test Cmax (g/mL) 6.02 1.56 0.37 25.98 Tmax (h) 1.22 0.80 0.19 65.40 AUC0t (g.h/mL) 30.54 7.57 1.78 24.78 AUC0 (g.h/mL) 31.36 7.70 1.81 24.55 t1/2 (h) 7.73 1.64 0.39 21.22 Kel (h1) 0.09 0.03 0.01 26.49

0.11 0.03 0.01 29.95

Cmax: Maximum measured plasma concentration; T max: Time of maximum measured plasma concentration; AUC0t: The area under the plasma concentrationversustimecurvefromtimezerotothelastmeasurableconcentration;AUC0:Theareaundertheplasmaconcentrationverses time curve from zero to infinity; t 1/2: Time required for the plasma drug concentration to decrease by one half; Kel: Apparent first order elimination or terminal rate constant; SEM: Standard error of mean; %CV: Coefficient of variation; Test: Troikaa Pharmaceuticals Ltd. India, Reference:McNeil,US. Table2:Pointestimateand90%confidenceintervalsfortheratiooftheproductsaveragesofTestandreferenceformulations Parameter Cmax AUC0t AUC0 Pointestimate test:reference 0.98 0.981 0.990 Lowerconfidencelimit 89.9 92.31 91.58 Upperconfidencelimit 109.9 104.1 106.6

Cmax: Maximum measured plasma concentration; AUC0t: The area under the plasma concentration versus time curve from time zero to the last measurableconcentration;AUC0:Theareaundertheplasmaconcentrationversestimecurvefromzerotoinfinity Pharmacokineticandstatisticalanalyses Maximal plasma concentration (Cmax) and time to reach the peak concentration(Tmax)wereobtaineddirectlybythevisualinspection ofeachsubject'splasmaconcentrationtimeprofile.Theslopeofthe terminal loglinear portion of the concentrationtime profile was determined by leastsquares regression analysis and used as the elimination rate constant (Kel). The elimination halflife was obtainedfromtheformula,t1/2=ln(2)/KelTheAUC0tfromtimezero to the last quantifiable point (Ct) was calculated using the trapezoidalruleandtheextrapolatedAUCfromCttoinfinity(AUC0 ) was to be determined as Ct/ Kel. The area under the plasma concentrationtimefrom0toinfinity(AUC 0)wascalculatedasthe sumoftheAUC0tplustheratioofthelastmeasurableconcentration totheeliminationrateconstant. To test the bioequivalence of the test and reference formulations, analysis of variance (ANOVA) for the crossover design was conducted on logtransformed Cmax, AUC0t, and AUC0. The formulations were to be considered bioequivalent if the log transformedratios(test/reference)ofCmax,AUC0t,andAUC0were within the predetermined bioequivalence range of 80% to 125% andifPwas>0.05forthe90%confidenceintervals.7 Safetyandtolerability General clinical safety was assessed via physical examinations and vital signs conducted at screening and at the end of the study. ClinicallaboratorytestsandECGswerealsoconductedatscreening, before dosing within each treatment period, and at the end of the study.Adverseeventswereassessedforseverityandrelationshipto treatmentthroughoutthestudy. RESULTS Pharmacokineticanalysis Themeanserumconcentrationtimecurvesof2ERformulationsof paracetamol products each administered as a single 650 mg oral doseto18healthyIndianmalevolunteersareshowninthefigure1. TheprimaryPKparametersforbothformulationsarelistedinTable 1.Themean(SD)Cmaxvaluesofthetestandreferenceformulations were 6.02 (1.56) and 6.17 (1.47) g/mL, respectively. The mean (SD) Tmax values were 1.22 (0.8) and 1.06 (0.72) hours. Results for theextentofabsorption,asdeterminedfrommean(SD)AUC 0t and AUC0 values, were 30.54 (7.57) and 31.36 (7.70) g/mL/h respectively,afteradministrationofthetestformulation;and31.12 (8.02) and 31.69 (8.12) g/mL/h after administration of the referenceformulation.Themean(SD)t1/2was7.73(1.64)hoursfor the test formulation and 7.0 (2.10) hours for the reference formulation.OnANOVA,noperiod,formulationorsequenceeffects wereobservedforanyPKproperty.The90%confidenceintervalsof the ratios (test vs reference) for the natural log (ln)transformed Cmax, AUC0t, and AUC0 are shown in Table 2 and summary statisticsareshowninTable3.The90%confidenceintervalsforthe ratiosofCmax,AUC0t,andAUC0were89.9to109.9,92.31to104.1 and91.58to106.6respectively,meetingthepredeterminedcriteria forbioequivalence.

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Dhaneshwaretal. IntJCurrPharmRes,Vol2,Issue4,2831 Safetyandtolerability adverse events were recorded, and there were no clinically significant changes in vital signs, clinical laboratory variables, ECG All 18 subjects completed the study and there were no premature parametersorphysicalexaminationfindingsduringthestudy.There withdrawals, replacements or death during the study. No serious werenoadverseeventsreportedduringthestudy. Table3:Summarystatisticsofparacetamolin18adult,subjectsunderfastingconditions. Parameters Summarystatistics GeometricMean LeastSquareMean(LSM) LSMRatioB/A% 90%ConfidenceInterval:B/A LowerLimit UpperLimit pvalue(ANOVA) Period Formulation Sequence IntrasubjectVariability:CV(%) Product Test Reference Test Reference Cmax (g/mL) 5.47 5.90 5.47 5.90 92.7 89.9 109.9 >0.05 >0.05 >0.05 17.45 AUC0t (g.h/mL) 28.68 29.27 28.68 29.27 98.0 92.31 104.1 >0.05 >0.05 >0.05 10.5 AUC0 (g.h/mL) 29.46 29.82 29.46 29.82 98.8 91.58 106.6 >0.05 >0.05 >0.05 28.1

A: Reference Product; B: Test Product; ANOVA: Analysis of variance; B/A: Bioavailability ratio Test (B) / Reference (A); %CV: Coefficient of variation.

Fig.1:linearplotofmeanparacetamolconcentrationversustimein18malesubjectsunderfastingconditions DISCUSSION This study examined the PK properties and bioequivalence of 2 formulations of paracetamol ER a newly developed extended releasebilayertabletandanestablishedbrandedtabletinhealthy Indian adult male volunteers. The 90% confidence intervals were completelycontainedwithin thepredefined bioequivalence criteria of 80% to 125% for the primary end point of Cmax and AUC. The studyresultsrevealedthatthe2formulationsofparacetamolwere similar in PK characteristics among these healthy Indian male volunteers. The 90% confidence intervals for the ratios of Cmax, AUC0t,andAUC0were89.9to109.9,92.31to104.1and91.58to 106.6 respectively, meeting the predetermined criteria for bioequivalence.Themeant1/2obtainedinthisstudywas7.73hours for the test formulation, which was comparable to that of the reference formulation at 7.0 hours. The mean Cmax of the test was 6.02 g/mL, which was comparable to that of the reference formulation6.17g/mL. The ER formulation of paracetamol and immediate release (IR) paracetamol have been reported to be clinically and statistically equivalent.Bothformulationsweresimilarintermsofbothonsetof analgesiaandpeakanalgesiceffect.6 The ER paracetamol taken three times daily was reported to be statisticallyandtherapeuticallynoninferiortoIRparacetamoltaken fourtimesdailyinpatientswithkneepainduetoosteoarthritis.The ER paracetamol may thus, be more convenient for patients with chronic pain and has the potential to enhance compliance and therefore pain relief.8 It may be of benefit to the patient at night time.6 Paracetamol is very well tolerated. Systematic reviews have found the rate of adverse events following its administration is not significantly different to that following administration of placebo,3,9,10whilehypersensitivityreactionsarerare.3,11 Although the major concern with paracetamol administration relates to the potential for hepatotoxicity, this is extremely rare following therapeutic dosing.3,12 In the present study both formulations were well tolerated and no adverse events were reportedduringthestudy. CONCLUSION In this study in healthy Indian adult male volunteers, a single 650 mg dose of the extend release bilayer formulation (test) of paracetamol met the regulatory criteria for bioequivalence to a single650mgdoseoftheestablishedtabletformulation(reference) basedontherateandextentofabsorption.Bothformulationswere welltolerated. ACKNOWLEDGEMENTS We are thankful to Troikaa Pharmaceuticals Ltd., Ahmedabad for providing the drug samples. We are also grateful to Ms. Rita Rana andotherclinicalstaffofPERDCentreforassistanceintheconduct ofpharmacokineticstudy. REFERENCES 1. Bertolini A, Ferrari A, Ottani A, Guerzoni S, Tacchi R, Leone S. Paracetamol: new vistas of an old drug. CNS Drug Rev 30

Dhaneshwaretal. IntJCurrPharmRes,Vol2,Issue4,2831 2006;12(34):25075. Portoles A, Puerro M, Terleira A, Rodriguez A, Caturla M C, Fernandez N, Vargas E. A new highabsorptionrate Paracetamol500mgformulation:acomparativebioavailability study in healthy volunteers. Curr Ther Res Clin Exp 2003; 64:401411. Oscier CD, Milner QJ. Perioperative use of paracetamol. Anaesthesia2009;64(1):6572. Rawlins M, Henderson D, Hijab JR. Pharmacokinetics of paracetamol after intravenous and oral administration. EuropeanJournalofClinicalPharmacology1977;11:2836. Ward B, AlexanderWilliams JM. Paracetamol revisited: a reviewofthepharmacokineticsandpharmacodynamics.Acute Pain1999;2:1409. CoulthardP,HillCM,FrameJW,BarryH,RidgeBD,BaconTH. Pain control with paracetamol from a sustained release formulation and a standard release formulation after third molar surgery: a randomised controlled trial. Br Dent J 2001;191(6):31924. US FoodandDrug Administration,CenterforDrug Evaluation and Research. Guidance for industry statistical approaches to establishing bioequivalence. http://www.fda.gov/cder/guidance/3616fnl.htm. Accessed on April6,2010. 8. Bacon,T.H.,Hole,J.G.,North,M.&Burnett.Analgesicefficacy ofsustainedreleaseparacetamolinpatientswithosteoarthritis of the knee. British Journal of Clinical Pharmacology 2002;53(6):629636. 9. Barden J, Edwards J, Moore A, McQuay H. Single dose oral paracetamol (acetaminophen) for postoperative pain. CochraneDatabaseofSystematicReviews2004;1:CD004602. 10. WeilK,HooperL,AfzalZ,etal.Paracetamolforpainreliefafter surgicalremovaloflowerwisdomteeth.CochraneDatabaseof SystematicReviews2007;3:CD004487. 11. Graham G, Scott K, Day RO. Tolerability of paracetamol. Drug Safety2005;28:22740. 12. Prescott LF. Therapeutic misadventure with paracetamol: fact or fiction. American Journal of Therapeutics 2000; 7:99114 7.

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