BIOEQUIVALENCE TESTING

Roger K. VERBEECK, Ph.D.

SCHOOL OF PHARMACY
UCL – Brussels
BELGIUM

[email protected]
 OUTLINE
 Concept of bioavailability
 Historical cases of bio-inequivalence
 Bioequivalence testing : EMA and FDA
guidelines
 Study designs
 Bioequivalence criteria
 Special issues
 A representative sample of the
slides that will be presented and
discussed during the course is
displayed hereafter.
 PHARMACOKINETICS

Temporal characteristics of drug effect and relationship to the

therapeutic window (e.g., single dose, oral administration).
 BIOAVAILABILITY

 The extent and the rate to which a drug

substance or its therapeutic moiety is
delivered from a pharmaceutical form
into the general circulation
 absolute bioavailability
 relative bioavailability
 ABSOLUTE BIOAVAILABILITY

AUCoral DOSEiv
F 
AUCiv DOSEoral
 RELATIVE BIOAVAILABILITY

 tablet A
□ tablet B
AUC formA
Frel 
AUC formB
 BIOAVAILABILITY

AUC1AUC2AUC AUC1=AUC2=AUC3
Cmax 3

Tmax

Changes in extent and/or rate of absorption influence the

drug plasma concentration-time profile and may therefore
affect the therapeutic efficacy
 BIOAVAILABILITY

formulation and physicochemical factors influencing drug

absorption and bioavailability
 BIOAVAILABILITY

 The practical importance of BA/BE testing has been

demonstrated by a number of clinical reports in the
60’s and the 70’s documenting medical problems due
to bio-inequivalence
 To allow prediction of the therapeutic effect the
performance of the pharmaceutical dosage form
containing the active substance should be known and
reproducible
 BIOAVAILABILITY

 Outbreak of phenytoin
intoxication around 1970 in
Queensland, Australia

 Calcium sulphate dihydrate in

the phenytoin capsules had
been replaced by lactose

Tyrer et al.: Outbreak of anticonvulsant intoxication in

an Australian city. Brit. Med. J. 4: 271-273, 1970.
 BIOAVAILABILITY

2.5 Burroughs Wellcome

Davis-Edwards
CONCENTRATION

2.0 APC batch B15703

APC batch 24928
SERUM

(ng/ml)

1.5

1.0

0.5

0.0
0 1 2 3 4 5 6
TIME (hours)

Lindenbaum et al.: Variation in the biologic availability of digoxin

from four preparations. New Engl. J. Med. 1344-1347, 1971.
 BIOEQUIVALENCE

 “Two medicinal products containing the same active

substance are considered bioequivalent if they are
pharmaceutically equivalent or pharmaceutical
alternatives and their bioavailabilities (rate and extent
of absorption) after administration in the same molar
dose lie within acceptable predefined limits. These
limits are set to ensure comparable in vivo
performance, i.e. similarity in terms of safety and
efficacy.”

GUIDELINE ON THE INVESTIGATION OF BIOEQUIVALENCE, EMA

(LONDON), 2010.
 BIOEQUIVALENCE TESTING

BE tests are not only required

for the registration of generic
drugs but are also carried out
during the development of
new drugs: e.g. during
scale-up and for post-approval
changes (SUPAC).
 IMMEDIATE RELEASE DOSAGE FORMS

http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2010/01/
WC500070039.pdf
 FDA Guidance for Industry

http://www.fda.gov/downloads/Drugs/
GuidanceComplianceRegulatoryInformation/Guidances/ucm072872.pdf
 EMA Guideline on the Investigation
of Bioequivalence, 2010

 STUDY DESIGN
- standard design: randomized, two-period,
two-sequence, single dose cross-over
design
- alternative designs: parallel design
(substances with very long half-lives) and
replicate designs (in case of highly
variable drugs or drug products)
 2 x 2 CROSS-OVER DESIGN

R period 1 period 2
A
N
D sequence 1 R W T
A
O
S
M
subjects H
I
O
Z
U
A
T sequence 2 T T R
I
O
N
 TWO-GROUP PARALLEL DESIGN

R
A
N group 1 TEST
D
O
M
subjects I
Z
A
T group 2 REFERENCE
I
O
N
 REPLICATE DESIGN
4-period, 2-sequence, 2-formulation design

period
R I II III IV
A
N
D sequence 1 T R T R
O
M
subjects I
Z
A
sequence 2 R T R T
T
I
O
N wash-out
 EMA Guideline on the Investigation
of Bioequivalence, 2010
 STUDY SUBJECTS
- the number should be based on an appropriate
sample size calculation ( 12),
- healthy volunteers to reduce variability,
- strict inclusion/exclusion criteria,
- subjects could belong to either sex,
- preferably non-smokers and without a history of
alcohol or drug abuse,
- patients, if the investigative active substance is
known to have serious adverse effects considered
unacceptable for healthy volunteers
 EMA Guideline on the Investigation
of Bioequivalence, 2010

 SAMPLING TIMES
o “The sampling schedule should
include frequent sampling around
the predicted tmax to provide a
reliable estimate of peak exposure.
In particular, the sampling schedule
should be planned to avoid Cmax
being the first point of the
concentration-time curve.”

Sampling times: 0, 0.5, 1, 2, 4, 6, 8 and 12 hours.

 EMA Guideline on the Investigation
of Bioequivalence, 2010
 SAMPLING TIMES

“The sampling schedule should also cover the plasma

concentration time curve long enough to provide a
reliable estimate of the extent of exposure which is
achieved if AUC0-t covers at least 80% of AUC0-.
At least 3 to 4 samples are during the terminal log-
linear phase in order to reliably estimate the terminal
rate constant, which is needed for a reliable estimate
of AUC0-.”
 EMA Guideline on the Investigation
of Bioequivalence, 2010

 PRIMARY PARAMETERS:

- AUC0-t or AUC0-72h: extent of absorption

- Cmax: extent and rate of absorption

- Tmax: rate of absorption
 RATE OF ABSORPTION
partial AUC
 For drug products where rapid absorption is of importance, partial AUCs can
be used as a measure of early exposure (FDA Guidance for Industry, 2003).
 The partial area can in most cases be truncated at the population median of
Tmax values for the reference formulation: AUCtmax
 EMA Guideline on the Investigation
of Bioequivalence, 2010

 BIOANALYTICAL METHODOLOGY

The bioanalytical part of BE trials should be

performed in accordance with the principles
of Good Laboratory Practice (GLP).
EMA draft Guideline on Validation of
Bioanalytical Methods, London, November
2009.
 EMA Guideline on the Investigation
of Bioequivalence, 2010

 BIOEQUIVALENCE CRITERIA:
are based on the calculation of a 90% confidence interval
according to the two one-sided tests procedure of Schuirmann

D.J. Schuirmann: A comparison of the two one-sided tests procedure

and the power approach for assessing the equivalence of average
bioavailability. J. Pharmacokinet. Biopharm. 15: 657-680, 1987.
 EMA Guideline on the Investigation
of Bioequivalence, 2010
 The pharmacokinetic parameters under consideration (e.g. AUC 0-t, Cmaxin case of a single dose
BE study) should be analysed using ANOVA.
 The data should be transformed prior to analysis using a logarithmic transformation.
 The terms to be used in the ANOVA model are usually sequence, subject within sequence, period
and formulation.
 A statistical evaluation of t max is not required. However, if rapid release is claimed to be clinically
relevant and of importance for onset of action or is related to adverse events , there should be
no apparent difference in median t max and its variability between test and reference product.
 90% CONFIDENCE INTERVAL

a 90% confidence interval has to be

calculated around the ratio of geometric
means obtained for AUC (and Cmax)
following administration of test and
reference preparation
 this ratio of geometric means is called
the point estimate
 EMA Guideline on the Investigation
of Bioequivalence, 2010

 For Cmax and AUC0-t the 90% confidence interval for the ratio of the test and
reference products should be contained within the acceptance interval of
80.00-125.00%.
 In specific cases of products with a narrow therapeutic range, the
acceptance interval may need to be tightened. Moreover, for highly variable
drug products the acceptance interval for Cmax may in certain cases be
widened (by using the reference scaled average bioequivalence approach).
90% CONFIDENCE INTERVAL

90% CI  1.04 (0.91 – 1.20)

point
estimate


0.80
 1.25
1.04
NARROW THERAPEUTIC INDEX DRUGS
EMA BE guideline - 2010

 In specific cases of products with a narrow therapeutic index (NTI),

the acceptance interval for AUC should be tightened to 90.00-
111.11%.
 Where Cmax is of particular importance for safety, efficacy or drug
level monitoring the 90.00-111.11% acceptance interval should also
be applied for this parameter.
 It is not possible to define a set of criteria to categorise drugs as
NTI drugs and it must be decided case by case if an active
substance is and NTI drug based on clinical considerations.
BIOPHARMACEUTICS CLASSIFICATION
SYSTEM (BCS)
permeability

dissolution

solubility

 solubility, dissolution and permeability are the 3

major factors controlling the oral absorption of drug
substances from IR oral medicinal products
BIOPHARMACEUTICS CLASSIFICATION
SYSTEM (BCS)

Class I Class II
High solubility Low solubility
High permeability High permeability

Class III Class IV

High solubility Low solubility
Low permeability Low permeability

Amidon et al.: A theoretical basis for a biopharmaceutic drug classification: the

correlation of in vitro drug product dissolution and in vivo bioavailability. Pharm.
Res. 12: 413-420, 1995.
BIOPHARMACEUTICS CLASSIFICATION
SYSTEM (BCS)

 the BCS was developed for regulatory

applications: to provide a basis for replacing,
in certain cases, in vivo BE studies by
equally or more accurate in vitro tests

 biowaiver: an acceptance for replacing an in

vivo BE study with in vitro dissolution testing