A RANDOMIZED, OPEN LABEL, BALANCED, TWOTREATMENT, TWO PERIOD, TWO SEQUENCE, SINGLE DOSE, TWO WAY CROSSOVER,ORAL ,BIOEQUIVALENCE

STUDY OF METOPROLOL SUCCINATE 50mg TABLET IN 24 HEALTHY ADULT HUMAN VOLUNTEER UNDER FASTING CONDITION

PRESENTED BY P VENKATA SREEDHAR REDDY 11Q31S0107

UNDER THE GUIDANCE OF Miss.RAMYA NAGASURI, M.Pharm Assistant Professor, Department Of Pharmacology , VCPRC, NELLORE Beloved Principal Dr. HARIBHASKAR, M. Pharm.,Ph.D. VCPRC, NELLORE.
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CONTENTS
INTRODUCTION AIM AND OBJECTIVE LITERATURE REVIEW DRUG PROFILE MATERIALS AND METHODOLOGY CLINICAL PHASES BIO ANALYSIS STATISTICAL METHODOLOGY RESULTS AND DISCUSSION CONCLUSION BIBILOGRAPHY
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INTRODUCTION
Why we need Bioavailability and Bioequivalence study?

Ensuring uniformity in standards of quality, efficacy and safety of pharmaceutical product.

Bioavailability can be generally documented by a systemic exposure profile obtained by measuring drug and/or metabolite concentration in the systemic circulation over time. Bioequivalence studies should be conducted for the comparison of two medicinal products containing the same active substance.
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Definitions: Bioavailability Bioavailability may be defined as the rate and extent (amount) of absorption of unchanged drug from it's dosage form . Bioequivalence: It is a relative term which denotes that the drug substance in two or more identical dosage forms, reach the systemic circulation at the same relative rate and extent i.e. their plasma concentration time profile will be identical with out significant statistical different .
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AIM AND OBJECTIVE

The aim of this study is to evaluate the Bioequivalence of Metoprolol Succinate 50mg extended release tablets in healthy, adult, human subjects under fasting conditions. To find out whether the test and reference products are biologically equivalent or not To find out bioequivalence of the Test (T) with Reference (R) product. To analyse Metoprolol Succinate content in human Plasma through bio-analytical method validation using LC-MS/MS.

To monitor the adverse events and to ensure the safety of the subjects.
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DRUG PROFILE AND CLINICAL PHARMACOLOGY

Drug Name Structure : : Metoprolol Succinate

Chemical Name

()1- (isopropylamino)-3-[p-(2methoxyethyl) phenoxy]-2propanol succinate (2:1) (salt) (C15H25NO3)2.C4H6O4 652.8 a white, crystalline powder Freely soluble in water & methanol AntiHypertensive, Antiangina, CCF
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Empirical formula : Mol.mass : colour : solubility : Therapeutic category :

PHARMACOKINETICS
Absorption Distribution

Metabolism
Elimination Pharmacology and Mechanism of Action: Metoprolol succinate is a betal-selective (cardio selective) adrenoceptor blocking agent, for oral blocking agent, for oral administration, available as extended release tablets. Metoprolol succinate extended release tablet has been formulated to provide a controlled and predictable release of Metoprolol for once daily administration.
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MATERIALS & METHODOLOGY

MATERIALS: Test formulation Reference formulation STUDY DESIGN Study Design Scheme table
Number of study
Sequence participants= N Period 1 Washout Period 2

Sequence I

24

Treatment A

7days

Treatment B

Sequence II

24

Treatment B

7 days

Treatment A
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STUDY FLOWCHART
SCREENING

ELIGIBILITY CRITERIA SUBJECT SECLECTION (n=24)

RANDOMIZATION TTREATMENT-T PERIOD-I WASHOUT -07 DAYS CROSS OVER TREATMENT-R

TREATMENT-R

PERIOD-II

TREATMENT-T

STUDY SAMPLE ANALYSIS (N24) PK & STATISTICAL ANALYSIS N24

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ETHICAL CONSIDERATIONS
Basic Principles Institutional Review Board Composition, Functions, and Operations Subject Selection :
Screening: The screening will be carried out after taking a written informed consent for screening from volunteers and will include the following: Demographic data, Medical history, Complete physical examination, Chest X-ray, ECG. Laboratory Parameter Investigations: Hematology, Blood grouping & Rh typing and bleeding time, Biochemistry, Hepatic profile Renal profile Urine Examination, Drugs of abuse on the day of study check-in of each period, Screening for infectious diseases.
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SUBJECT SELECTION
Inclusion Criteria: Subjects who will provided written consent form. Healthy males within the age range of 18 to 45 years Preferably Non- Smokers Willingness to provide written informed consent to participate in the study Body mass index of 18.5 kg/m2 and 24.9 kg/m2, with body weight not less than 50 kg Absence of significant disease or clinically significant abnormal laboratory values or laboratory evaluation, medical history or physical examination during the screening Have a normal 12-lead ECG or one with abnormality considered to be clinically insignificant Have a normal chest X-ray . Comprehension of the nature and purpose of the study and compliance with the requirement of the distributed ICF.
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SUBJECT SELECTION
Exclusion Criteria: Personal/Family history of allergy or hypersensitivity to the drug or allied drugs. Any major illness in the past 90 days or any clinically significant ongoing chronic medical illness e.g. Congestive Cardiac Failure (Heart failure), Hepatitis, Hypotensive episodes, Hyperglycemia etc., Presence of any clinically significant abnormal values during screening e.g. significant abnormality of liver function test, renal (kidney) function test etc., Severe cardiac, renal or liver impairment, gastro-intestinal disease or other conditions, any other organ or system impairment History of seizures, epilepsy or any kind of Neurological disorders Past history of Anaphylaxis or angioedema Presence of disease markers of HIV 1 and 2 and hepatitis B, C virus
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SUBJECT SELECTION
Exclusion Criteria: Consumed alcohol within 48 hrs prior to dosing Consumption of Xanthine containing derivatives (coffee, tea, cola drinks, chocolate) or tobacco products within 48 hours prior to dosing Use of any recreational drug or a history of drug addiction Participation in any clinical trial within the past 90 days History of difficulty with donating blood or difficulty in accessibility of veins in left or right arm Donation of blood (one unit or 350 ml) within 90 days prior to receiving the first dose of study medication Receipt of any other prescription drug or over the counter (OTC) drugs within two weeks prior to receiving the first dose of study medication or repeated use of drugs within the last four weeks An unusual diet for whatever reason e.g. low sodium diet, for two weeks prior to receiving any medication and throughout subjects participation in the study Recent history of dehydration from diarrhoea, vomiting or any other reason within a period of 24 hours prior to the study
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SUBJECT SELECTION
Withdrawal Criteria: Subjects may be withdrawn from the study by the principal investigator or co-investigators for any of the following reasons during the course of the study: If the subject suffers from significant illness If the subject requires concomitant medications which may interfere with pharmacokinetic of the study drug If the subject has entered the study in violation of the inclusion and the exclusion criteria If the subject is found to be non co-operative If the subject decides to voluntarily dropout from the study
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RANDOMIZATION
Subject No 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 Treatment Sequence RT TR TR RT` RT TR TR RT RT TR RT TR TR RT RT TR RT TR TR RT RT TR RT TR Period I R T T R R T T R R T R T T R R T R T T R R T R T Period II T R R T T R R T T R T R R T T R T R R T T R T R

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DRUG SUPPLIES, LABELING, ACCOUNTABILITY AND STORAGE

The sufficient quantities of test and reference products with product name, strength, number of dosage units, manufacturer, lot/batch no, expiry date and storage conditions and their CoAs and In-vitro dissolution data will be received from the sponsor, for the conduct of the study. The drug products will be stored in the pharmacy under prescribed storage conditions and will be logged-in the Drug Receipt, Dispensing and Accountability Log Book.

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DRUG DISPENSING & DOSING

Drug Dispensing:
The Pharmacist will use required number of drug products for dispensing in to the drug-dispensing containers as unit doses based on total number of study subjects to conduct each period. Remaining drug products will be stored in their original container as retention samples. The issued drug products will be transferred to the drug-dispensing containers as unit doses.

Dosing and Treatments:

The Principal Investigator/Clinical Investigator/Physician will verify the treatment as per the randomization schedule. A single oral dose of drug 50 mg x 1 tablet of Reference (R) or Test (T) product will be administered during each period by study personnel After an overnight fasting of at least 10 hours,. Subjects will receive the alternate treatment in the subsequent period after crossover with the following treatment sequence i.e. either R-T or T-R.
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DIETARY PLAN
Subjects will be provided standard meal (dinner) (Day 0), at dinner consisting of approximately 1000-1200 calories.

Standard meals comprising of 2200-2400 calories per day was provided at 4.00, 8.00 and 13.00 hours post-dose (i.e. lunch, snacks, dinner respectively) on Day 1 during both the periods
Drinking water will not be permitted one hour before dosing and until one hour post-dose, at other times drinking water will be permitted ad libitum.

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PK SAMPLING
A total of 16 blood samples (1 5 ml) were collected in pre-labeled K2 EDTA vacutainers in each period. The blood samples will be collected prior to administration of dose in each period at pre-dose (0.00) Two blood samples were withdrawn and a single sample each time at 0.33, 0.67, 1.00, 1.33, 1.67, 2.00, 2.50, 3.00, 4.00, 6.00, 9.00, 12.00, 16.00 and 24.00 hours post-dose.

Handling of Blood Samples:

After collection the blood samples were centrifuged at 4000 rpm for 10 minutes at 4C for separating the plasma. Centrifugation of all samples will be done within 30 minutes after each sample draw time point. All plasma samples in cryovials were separated and are aliquoted into two set(s) and are transferred to deep freezer ,maintained at -20C + 5C from 0.00 (pre-dose) to 12.00 hours post-dose, after which the samples were directly transferred to a deep freezer maintained at 70oC + 10oC.
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SAFETY MONITORING
Vital Signs: In each period, vital signs monitoring will be done in sitting posture (except respiration rate which will be taken in supine position) at the time of check-in, at pre-dose (0.0) and at 2.00, 6.00, 12.00, and 24.00

hours post dose.

Handling and Reporting of Adverse Event: No Adverse Event was Observed

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BIO ANALYTICAL PROCEDURE AND SAMPLE ANALYSIS

The bio analytical part of bioequivalence trials was conducted according to the applicable principles of Good Laboratory Practice (GLP). (EMEA / OECD GLP / WHO GLP STANDARD). Sample analysis: Samples were assayed for plasma drug using a validated method developed at the bioanalytical unit. The method was validated over a concentration range of 15.00 ng/mL to 3000.00 ng/mL for Metoprolol Succinate .

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DATA ANALYSIS
Data Entry: Pharmacokinetic Parameters and Analysis Pharmacokinetic parameters for plasma drug were calculated by using WinNonlin software. Tmax, Cmax, AUC0t, AUC0, Kel and T1/2 Statistical analysis Statistical analysis was performed on pharmacokinetic data of samples assayed and quantified for plasma drug using the SAS version 9.1.3 and WinNonlin 5.1 version software. The following statistical information was provided for Tmax, Cmax, AUC0t, AUC0 for drug: Geometric Mean , Ratio of Means, 90% Confidence Intervals. Analysis of Variance (ANOVA) The Untransformed and Lntransformed pharmacokinetic parameters (Cmax, AUC0-t and AUC0-) was statistically analysed using General Linear Model (PROC GLM) of SAS version 9.1.3 software
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Pharmacokinetic terms
C max :This is the maximum drug concentration achieved in systemic circulation following drug administration. T max :It is the time required to achieve maximum drug concentration in systemic circulation . AUC o-t : Area under the plasma concentration-time curve from 0 hr to the last quantifiable concentration to be calculated using the trapezoidal rule . AUC 0- : Area under the plasma concentration-time curve from 0 hr to infinity to be calculated as the sum of AUC o-t plus the ratio of the lest measurable concentrations to the elimination rate constant. K el : Apparent first-order terminal elimination rate constant calculated from a semi-log plot of the plasma concentration versus time curve . T 1/2 : Elimination half life of a drug is the time necessary to reduce the drug concentration in the blood , plasma, or serum to one-half after equilibrium is reached.
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Pharmacokinetic data
The details of the ratios of means of test and reference parameters such as In-transformed Cmax, AUC0-t, AUC0- of Test and Reference at 90% confidence interval were as follows: 90% C.I. for log transformed data Metoprolol Cmax 106.93 - 118.03

Parameters

AUC0-t
AUC0-

96.02 - 111.95
91.54 - 109.25
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RESULTS AND DISCUSSION

PK Parameters T
Tmax (hr)
Cmax(ng/mL) AUC0-t (ng.h/mL) AUC0- (ng.h /mL) T (hr) Kel(1/hr) AUC_% Extrap_Obs
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Metoprolol Mean ( S.D.) R

9.506 (3.8720) 20.5371 (9.97919) 407.7738 (260.24935) 461.4856 (320.91953) 7.645 (3.5970) 0.1049 (0.03506) 12.2143 (12.19323)
11/3/2013

5.500 (3.8814) 22.6539 (9.93367) 417.8271 (271.10889) 457.8906 (315.61618) 7.358 (2.3436) 0.1025 (0.02980) 9.6239 (8.74828)

RESULTS AND DISCUSSION

Comparative Linear Plot of Mean Plasma Concentration of Metoprolol Succinate Versus Time in Healthy Adult Human Subjects ( N=24), under Fasting Conditions Metoprolol Succinate _ Linear Mean Plot

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RESULTS AND DISCUSSION

Comparative Semilogrithmic Plot of Mean Plasma Concentration of Metoprolol Succinate Versus Time in Healthy, Adult, Human Subjects ( N=24), under Fasting Conditions Metoprolol Succinate Semi log Mean Plot

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RESULTS AND DISCUSSION

The test product, Metoprolol Succinate 50 mg tablet is bioequivalent with the reference product, TOPROL- XL 50 mg tablet (contains Metoprolol Succinate 50 mg ) in terms of rate and extent of absorption under fasting conditions. The ratios for geometric least square means lie with in the acceptance ranges 80-125 % for log transformed C max , AUC 0-t and AUC 0-inf. The 90% confidence interval for the ratio of geometric least square means values were within the acceptance limit of 80%- 125% for Log transformed Cmax , AUC 0-t and AUC 0-inf for Metoprolol Succinate . The results obtained for Metoprolol Succinate were, lower limites 106.93,96.02,91.54 and upper limits 118.03,111.95,109.25.C max, AUU0-t and, AUC0-, respectively.
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CONCLUSION
All the study procedures followed were in compliance with the protocol and the ICH-GCP guidelines, Declaration of Helsinki and Schedule Y. From the clinical data it can be concluded that the study objectives like the safety and efficacy of the test product has been achieved. Based on clinical, pharmacokinetic and statistical data obtained from 24 healthy, adult, male, human subjects under fasting conditions, it was concluded that a single dose of test formulation T containing drug Metoprolol Succinate 50 mg was found to be safe and bioequivalent to the reference formulation R TOPROL-XL containing Metoprolol Succinate 50 mg .
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BIBLIOGRAPHY
Guidance for Industry. Bioavailability and Bioequivalence Studies for Orally Administered Drug Products . General Considerations. U.S. Department of Health and Human Services.Food and Drug Administration ,Center for Drug Evaluation and Research (CDER) March 2003 . Guidelines for bioavailability & bioequivalence studies, Central Drugs Standard Control Organization, Directorate General of Health Services, Ministry of Health & Family Welfare, Government of India, New Delhi. (March 2005). http://en.wikipedia.org http://www.drugs.com/ http://www.FDA.gov

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BIBLIOGRAPHY
Brahmankar.D.M, Sunil, B. Jaiswal , Biopharmaceutics And Pharmacokinetics, Published by M.K. Jain for Vallabh Prakashan ,Delhi110034,1st Edition reprint 2007,P.282. Chobanian AV, Bakris GL, Black HR et al. (December 2003). "Seventh report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure". Hypertension 42 (6): 120652.doi:10.1161/01.HYP.0000107251.49515.c2. PMID 14656957. Carretero OA, Oparil S (January 2000). "Essential hypertension. Part I: definition and etiology". Circulation 101 (3): 329 35. doi:10.1161/01.CIR.101.3.329. PMID 10645931. Fisher ND, Williams GH (2005). "Hypertensive vascular disease". In Kasper DL, Braunwald E, Fauci AS, et al.. Harrison's Principles of Internal Medicine (16th ed.). New York, NY: McGraw-Hill. pp. 1463 81. ISBN 0-07-139140-1. Palatini P, Julius S (June 2009). "The role of cardiac autonomic function in hypertension and cardiovascular disease". Curr. Hypertens. Rep. 11 (3): 199205. doi:10.1007/s11906-009-0035-4. PMID 19442329
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