PHAR 3352 Pharmacokinetics and Biopharmaceutics

Bioequivalence Study Presentation

Article 6 Bioequivalence Study of Amlodipine Maleate versus Amlodipine
Besylate in Healthy Volunteers
NUR AQILAH BINTI ADNAN 2010-2580
ALIAH NUR SAADAH BINTI SHARIL 2010-2405
ENUSHA DEVI A/P RAMESH 2010-2487
NUR QISTINA HUMAIRA
2010-2493
EE XUAN RU
ANITH SYAHIRAH YAAKUB 2010-2848
YEONG MIN YUE 1908-1028
BAHRI OMAR 2010-2572
2010-3016
 Introduction
❖ Amlodipine is a long-acting dihydropyridine type Ca2+ channel blocker
(CCB).
❖ Main therapeutic indications include essential hypertension,chronic stable
angina, vasospastic angina, myocardial ischemia, and Raynaud’s disease.
❖ Administration of amlodipine relaxes vascular smooth muscle, induces
decreased after load, decreases systemic blood pressure and increases coronary
blood flow.
❖ Amlodipine was developed by Pfizer and is marketed primarily as a besylate
salt.
❖ Other salts of amlodipine were developed and are used in clinical practice.
 Subjects
Subjects (24) ● 18–45 years of age;
● The mean weight for males was 71.5 ± 5.3 Kg, and 56.4 ± 3.9 Kg for
females
● Good general clinical condition, as assessed by clinical history, clinical
examination, and electrocardiogram (ECG), and with results within the
normal range of laboratory values for hematology, clinical chemistry, and
urinalysis;
● Negative test results for serum hepatitis B surface antigen, hepatitis C
antibody, and human immunodeficiency (HIV) antibody;
● Capability of understanding purposes and requirements of the study.
 Drugs
10 mg tablets containing amlodipine were used:

Experimental Drug Reference Drug

Amlodipine maleate, batch Amlodipine besylate, Norvasc®

number S013/7, certified by 10 Pfizer Italia S.p.A. batch No.
Omicron Pharma S.r.l. No. 510005531I, expiration date
359/05 01/2010.
Study Design & Procedures
❖ Single-dose, randomised, two-period crossover design
❖ Open for investigators, but blind for the analyst
❖ Screening period of 40 days
❖ Amlodipine maleate or besylate was administered orally once to each
individual
❖ A balanced random number generator was used to create a randomization
schedule that determined the order of administration
 Study Design & Procedures
12 Volunteers
(6 men & 6 Women)

Amlodipine besylate Amlodipine maleate

10 mg 10 mg

8.00 a.m. after fasting 10 hr

Amlodipine maleate Amlodipine besylate

10 mg 10 mg

12 Volunteers 14 days
(6 men & 6 Women)

❖ Food was served 5-9 hours after the dose.

❖ Grapefruit juice, alcohol, caffeine were not allowed 48 hours before the dose till the last blood sample was taken
❖ No medication was permitted throughout the trial period unless it was necessary to treat an adverse event.
❖ Blood was taken at 0, 1, 2, 4, 5, 6, 8, 10, 12, 24, 48, 72, 120, and 144 hours.
❖ Test tubes with blood sample were centrifuged at 1,000 g for 10 min at 20°C then kept at -30°C for analysis.
Analytical Methods
Pharmacokinetics & Statistics Analysis
AUC & Cmax - Detects 20% differences in the Pk parameters
- Power = 80%
- Alpha error = 0.05

Laboratory data Microsoft Excel worksheet

Analysis PCModfit (version 3.0), EquivTest (version 2.0), Origin (version 7.0)

Validation Procedures included in the distribution kit

Continuous parameters - Logarithmic normal distribution

- Interval estimators including geometric mean and exponential function of mean ±
standard deviation, and median, min, and max values

Non-continuous parameter parameter of the time to attain peak of plasma concentrations (tmax), only median,
min, and max values were reported

❖ Two-way analysis of variance (ANOVA) for Latin square design was used to evaluate differences
between treatments, periods, and subjects.
❖ For the non-continuous parameter (tmax), the non-parametric Hodges-Lehmann test was used to
compare paired data of the two treatments.
Pharmacokinetics & Statistics
❖ Area under the curve (AUC) & Peak plasma concentration (Cmax) =
detection of 20% differences in the pharmacokinetic parameters between the two formulations, with
a power of 80% and an alpha error of 0.05.
❖ All data from laboratory were transferred into Microsoft Excel worksheet.
❖ Analysis of data : PCModfit (version 3.0), EquivTest (version 2.0), and Origin (version 7.0).
❖ Validation of programs were according to procedures included in the distribution kit.
❖ For continuous parameters : A logarithmic normal distribution was assumed together with interval
estimators including geometric mean and exponential function of mean ± standard deviation, and
median, min, and max values.
❖ For the non-continuous parameter, only median, min, and max values are reported.
❖ Two-way analysis of variance (ANOVA) for Latin square design was used to evaluate differences
between treatments, periods, and subjects.
❖ For the non-continuous parameter (tmax), the non-parametric Hodges-Lehmann test was used to
compare paired data of the two treatments.
Amlodipine Maleate versus Amlodipine Besylate
 Test of Bioequivalence
Statistical analysis European Agency for the Evaluation of Medicinal Products (EMEA)

Pk parameters International regulatory authorities

Cmax & tmax Plasma concentration–time data

AUCt (0-144 hours) Linear trapezoidal rule

AUCinf AUCinf = AUCt + Ct /Ke

t½ t½ = ln2/Ke

❖ To determine whether these two drugs are pharmacokinetically equivalent, the calculated individual
Cmax, AUCt, and AUCinf, as well as their ratios (test/reference), using log transformed data were assessed.
❖ The drugs were considered pharmacokinetically equivalent if the difference between the compared
parameters was statistically not significant (p < 0.05) and the 90% confidence limit for these parameters fell
within 0.8 to 1.25.
❖ These limits were broadened to 0.75 to 1.33 for Cmax, according to EMEA, in view of the high
within-subject variability and in absence of toxicity associated with Cmax .
 Tolerability Assessment
❖ Blood pressure and heart rate were measured using an automatic monitor (Mortara Rangoni,
Italy) in supine position at 3, 6, and 9 h
❖ ECG was done at 8 h post-dose.
❖ To assess tolerability profiles of the two salts of amlodipine, hematological cytometric analysis,
lipids profile, electrophoretic analysis of proteins, and tests for hepatic and renal function were
performed at baseline and at the end of the study.
❖ Adverse events (AEs) were also monitored and recorded on case-report forms based on
patient interview and physical examination.
Results
Plasma Level
 Results
Amlodipine Maleate Amlodipine Besylate

AUC Range (ng*h/g) 170.88 - 446.37

AUC Mean Values (ng*h/g) 304.69 320.41

Cmax Range (ng/g) 5.04-10.67 3.40-10.67

Inter-individual Coefficient 22.94 21.73

(%)
 Results
Bioequivalence Evaluation
Using Westlake’s 90% confidence interval:
- AUC: both formulation shows confidence
limits between the acceptance limits of 0.80
and 1.80
- Cmax: both formulation shows confidence
limits between the acceptance limits of 0.75
and 1.33
Using Schuirmann’s two one-sided t-tests
procedure
- The p value observed is below 0.05 which
consider as accept to the bioequivalence for
the formulations
Tolerability
1. 6h post-dose there is a report on elevation of
heart rate for both formulations
- 8.5beats/min → amlodipine maleate
- 8.3beats/min → amlodipine besylate
2. 11 adverse effect reported during the study
which probably or possibly due to the treatment
- 4 after exposed to the amlodipine maleate
- 7 after given amlodipine besylate
- AE such as dizziness and tiredness
→ Not much different in terms of AE between
amlodipine maleate and amlodipine besylate
 Conclusion
All recognize pharmacokinetically equivalent two drugs if the
difference of compared parameters is statistically not significant (p <
0.05) within the 90% confidence interval, following this statement
and according to clinical result obtained from amlodipine plasma
levels all measuring criterias above do not have that significant
difference, These findings suggest that the two salts of amlodipine
investigated (maleate and besylate) may be equivalent in the
therapeutic indications of the drug.