Bioequivalence Studies

DR. ASIM KUMAR MITRA

 What and Why?
• What?
– Bioavailability is defined as the rate and extent
to which the active drug ingredient is absorbed
and becomes available at the site of drug action
– Two drug products are said to be bioequivalent if
they are pharmaceutical equivalent or
pharmaceutical alternatives, and if their rates
and extents of absorption do not show a
significant difference.
 What and Why?
Fundamental Bioequivalence
Assumption

When a generic drug is claimed

bioequivalent to a brand-name drug, it is
assumed that they are
therapeutically equivalent.
 Points to Clear
¨ Bioavailability – rate and extent at which a drug
substance... becomes available in the general
system (product characteristic!)

¨ Bioequivalence – equivalent bioavailability

within pre-set acceptance ranges

¨ Pharmaceutical equivalence  Bioequivalence

¨ Bioequivalence  Therapeutic equivalence

 Generic Drug: Definition

• Same active ingredient (s)

• Same route of administration
• Same dosage form
• Same strength
• Same indications
Compares to reference listed drug (RLD)
 Bioequivalence (BE): Definition
“the absence of a significant difference in the rate
and extent to which the active ingredient or active
moiety in pharmaceutical equivalents or
pharmaceutical alternatives becomes available at
the site of drug action when administered at the
same molar dose under similar conditions in
an appropriately designed study.”

CDER U.S. Food & Drug Administration

 Bioequivalence
90
80
Concentration (ng/mL)

70
60
Test/Generic
50
Reference/Brand
40
30
20
10
0
0 5 10 15 20 25 30
Time (hours)
 Goals of BE
Bioequivalence studies focuses the impact of
changes to the dosage form to ensure that the
product is comparable to the standard
product available in the market

• Establish that a new formulation has therapeutic equivalence

in the rate and extent of absorption to the reference drug
product.
• Important for linking the commercial drug product to clinical
trial material at time of NDA
• Important for post-approval changes in the marketed drug
formulation
Why do we need Bioequivalence
studies?
 No clinical studies have been performed in
patients with the Generic Product to support
its Efficacy and Safety.
 With data showing similar in vivo
performance (= Bioequivalence)
Efficacy and Safety can be extrapolated from
the Innovator Product to the Generic
Product.
Bioequivalence

Bioequivalence

Bioavailability

Pharmaceutical
equivalent

Pharmaceutical
alternatives
 Bioequivalence

Two medicinal products are bioequivalents if

they are pharmaceutical equivalents or alternatives
and if their bioavailabilities (rate and extent) after
administration in the same molar dose are similar
to such degree that their effects, with respect
to both efficacy and safety, will essentially be
the same.
 Bioequivalence
Therapeutic equivalence of a multiscource product can
be assured when the multiscource product is both
pharmaceutically equivalent/alternative and
bioequivalent.

Concept of interchangeability includes the

equivalence of the dosage form as well as for the
indications and instructions for use.

TE = PE + BE
 Bioequivalence
• Pharmaceutical equivalent does not necessarily
imply therapeutic equivalence:

- difference excipients
drug
- difference manufacturing process
performance?
- other variables
 Bioequivalence
• Therapeutic equivalent does not necessarily imply
bioequivalence:

- sensitivity
- different formulations equivalence?
- different active substance
 Bioequivalence
Important PK parameters

Cmax:
the observed maximum concentration
of a drug  measure of the rate of
absorption
AUC:
area under the concentration-time
Curve  measure of the extent of
absorption tmax:
time at which Cmax is observed
 measure of the rate of absorption
Plasma concentration time profile

Cmax
AUC

Tmax
 Bioequivalence – single dose
Basic design considerations:

minimize variability not

attributable to formulations

minimize bias

goal: compare performance

 Bioequivalence – single dose
Golden standard study design:

single dose, two-period,

crossover

healthy volunteers

Reference (comparator)/
Test (generic)
 Study Design: Basic design consideration

• Minimize variability not attributable to

formulations
• Minimize bias

To compare performance of two products!!!

 Study Designs
• Single-dose, two-way crossover, fasted
• Single-dose, two-way crossover, fed

Alternative
• Single-dose, parallel, fasted (Long half-life)
• Single-dose, replicate design (Highly Variable
Drugs)
• Multiple-dose, two-way crossover, fasted
(Less Sensitive, non-linear kinetic)
 Study Designs

• Duration of washout period for cross-over

design
- should be approximately > 5 times the
plasma apparent terminal half-life
- However, should be adjusted accordingly for
drugs with complex kinetic model
 Planning a BE Study
Ethical Considerations

¨ An independent body of medical, scientific and non-

scientific members
¨ Responsibility is to ensure the protection of the
rights, safety and well-being of human subjects
involved in a trial by,
¨ Among other things, reviewing, approving, and
providing continuing review of trial protocol and
amendments and of the methods and material to
be used in obtaining and documenting informed
consent of the trial subjects;
 Planning a BE Study
Ethical Considerations
Composition requirements ICH GCP
¨ At least 5 members
¨ At least one member whose primary area of
interest is a non-scientific area
¨ At least one member who is independent of the
trial site
¨ Members without conflicting interest

 Only those members independent of the investigator and the

sponsor should review on a trial-related matter
 Planning a BE Study
Ethical Considerations
e.g. additional US FDA requirement for
IRB composition:

¨ Diverse backgrounds (race, gender, cultural,

qualification)
¨ Not entirely one gender
¨ Special expertise may be invited but without
voting rights
 Planning a BE Study
Ethical Considerations
Required documents

¨ Protocol (signed at least by the principal

investigator)
¨ Patient Information Sheet/Consent Form
¨ Investigator´s Brochure
¨ Subject recruitement procedures (e. g.
advertisements)
 Planning a BE Study
Ethical Considerations
Approval notification to Investigator
¨ Timely written approval
- Identification of study (title, protocol number, version, investigator,
site)
- Specify all items reviewed
- Date & place of review
- Trial/study related decisions
- Reasons for modifications & disapprovals
Minimum information required by ICH-GCP:
¨ Date of the meeting
¨ Documents reviewed (versions & dates)
¨ List of members
 Planning a BE Study
Study Protocol

♦ „A document that describes the objective(s),

design, methodology, statistical
consideration and organisation of a trial. It
usually gives the background and rationale of
the trial …“
Ref.: ICH GCP Guidance
 Planning a BE Study
Study Protocol
General Information/Title Page
♦ Title
♦ Protocol Number
♦ Version Number/Date
♦ Sponsor Details
♦ Name, Address, Telephone
♦ Monitor/Medical Personnel
♦ Investigator Details
♦ Principal Investigator, Medical Doctor
♦ Other Laboratory/Institution Details

 Responsibilities!
 Planning a BE Study
Study Protocol
Protocol Development

Definition of Responsibilities

¨ Organisation, premises, personnel & QMS

¨ Clinical phase
¨ Bioanalytical phase
¨ Statistics and reporting
¨ Archival
 Planning a BE Study
Protocol Development
Drug substance / Drug products

basic knowledge about particularities e.g.

¨ pharmacokinetics (t1/2, peak concentration,

metabolism…)

¨ practicability of roughly anticipated

measurement period and/or wash-out period
(crossover study possible?)
 Planning a BE Study
Protocol Development
Drug substance / Drug products

basic knowledge about particularities e.g.

¨ important side effects (acceptable for healthy

volunteers?)
 Planning a BE Study
Protocol Development
Drug substance / Drug products
basic knowledge about particularities e.g.

¨ concept of bioanalytical method available?

¨ plasma concentrations sufficiently

quantifiable (LOQ) (administration of more than one
dosage form necessary/possible?)
 Planning a BE Study
Protocol Development
Drug Products
¨ Availability
¨ Certification
¨ Content
¨ In vitro dissolution
¨ Preparation of investigative products per volunteer
acc. to GMP

F Protocol amendment for product details frequently

necessary
(e. g. labeling)
 Planning a BE Study
Study Subjects
¨ Selection of subjects
♦ description of volunteers; smoker, vegetarian,
phenotyping….
♦ verifying health of volunteers ( e. g. ECG, clinical blood
chemistry, blood pressure…)
♦ number of volunteers depending on variability; at
least 12 (EU: healthy, 18-55y; FDA: both sexes, > 18y)
♦ randomisation
objective: minimising interindividual variability in order to
detect product differences!
 Planning a BE Study
Study Subjects
¨ Selection of subjects
¨ participation of healthy volunteers
¨ reasonable inclusion and exclusion criteria
¨ comprehensive verbal and written information
and informed consent
¨ volunteers´ insurance
¨ reimbursement
 Planning a BE Study
Study Subjects
¨ Number of subjects

¨ Required sample size depends on variability

either known through reasonable literature or by
means of a pilot study
¨ “low” variability: ~ 12 – 20 volunteers
¨ “high” variability: ~ 24 – 26 (plus) volunteers
 Planning a BE Study
Study Subjects
¨ Number of subjects ctd.

¨ Required sample size depends on the expected

mean difference between the test and reference
formulation

F For sample size calculation see literature data (e.g. Eur J Drug Metab
Pharmacokinet 30 (2005) 41; J Biopharm Stat 13 (2003) 529; Stat Med 18
(1999) 93 …)

¨ Consideration of possible withdrawals

 Planning a BE Study
Study Subjects
¨ Subject withdrawals
¨ subject must adhere to study requirements but …

¨ they are free to break off at any time

¨ definition of “drop-outs” in the protocol (reason,
reimbursement policy, handling of data, follow-
up…)
¨ concomitant medication
¨ reporting
 Planning a BE Study
Study Design
¨ Crossover-design
“latin square” / balanced / randomized

F Intra-individual comparison!
¨ Parallel group design
¨ Replicate design
 Planning a BE Study
Standardisation
¨ Procedure of drug intake
¨ time of administration (fasted or fed state)
¨ liquid volume
¨ traceability of administrations

¨ cave: e.g. granules, suspensions liquid formulations!

(require ‘method sheet’)
 Planning a BE Study
Standardisation
¨ Standardized fluid and food intake (time,
composition, amount)
¨ Prohibition of alcohol
¨ Restriction of xanthins (coffee*, coke, chocolate,
chewing gum, grapefruit….)
¨ Standardized posture
¨ Restriction of physical activities
…
*cave: withdrawal may cause headache
 Planning a BE Study
Standardisation
¨ Fasted state

¨ Confinement of subjects at least 10 h prior to

drug administration
¨ Last food intake ~10 h prior to drug intake
¨ No food or fluids ~2 h prior to drug intake
¨ Drug administration with ~150-200 ml (e.g.)
water
¨ Light standardized meal not before ~4 h post-
dose
 Planning a BE Study
Standardisation
¨ Fed state

¨ Define time of drug administration and

food intake, (e. g. drug intake within 30 min. before,
immediately before or after the standardised meal)

¨ High fat meal may serves to investigate the

„worst case“ scenario
 Planning a BE Study
Study Samples
♦ Sampling
♦ number of samples
♦ sampling times (Cmax!)
♦ time of sampling (extrapolated AUC max. 20 %)
♦ wash-out-phase (4 – 5 half-lifes)

 knowledge of basic pharmacokinetics of the particular

drug substance is inevitable!

objective: characterisation of ‚drug input‘!

(see e.g. sect. 3.1 of the EU guidance 1401/98)
 Planning a BE Study
Study Samples
¨ Number of samples

¨ sufficient to “describe” at least 80 % of

total AUC

¨ usually ~12– 18 samples (minimum)

 Planning a BE Study
Study Samples
¨ Sampling times

¨ appr. 3 – 4 to describe drug “input”

¨ appr. 3 sampling times around peak
concentration
¨ appr. 3 – 4 to describe elimination

 Minimum!
 Planning a BE Study
Study Samples
¨ Wash-out-phase

¨ must be long enough to avoid residual

concentrations
¨ closely related to the limit of
quantitation
¨ metabolites may be considered
 Planning a BE Study
Sampling
¨ Blood withdrawal equipment (consider
bioanalytical method)
¨ Preparation of plasma or serum
¨ cooling
¨ centrifugation
¨ aliquotation
¨ labeling
¨ freezing
¨ transport…
 Planning a BE Study
Bioanalytical Method
¨ The protocol should state

¨ the bioanalytical method/detection

¨ the limit of quantitation (1/10 of the
expected peak concentration should be
measurable)
¨ the validation concept
¨ whether metabolites are to be considered
 Planning a BE Study
Calculations

¨ The protocol should state (-among others-)

¨ the transfer of bioanalytical results for

biostatistical calculations
¨ the handling of missing data

¨ the handling of digits

 Planning a BE Study
Modified Release Products
¨ The protocol should state/ensure/consider

¨ direct switching vs. wash-out

¨ primary characteristics (e.g. AUCtau, Cmax,

Cmin…)
¨ consideration of fluctuation (e.g. Ptf…)

¨ compare Cmin to ensure steady-state

 Planning a BE Study -
Adverse Events
¨ Definitions and handling/information

¨ Evaluation of seriousness
¨ Evaluation of relation to investigative
drugs

¨ Treatment (cave: concomitant drug intake should be tested ‘a

priori’ for possible analytical interferences)
Planning a BE Study-
Results…
 Guidelines for BE-Studies

1996: ICH GUIDELINES

2000: REVISION OF ICH GUIDELINES
2001 : INDIAN GCP GUIDELINES
2003 : REVISION OF BE GUIDELINES
2005 : REVISION OF SCHEDULE Y
 GCP guidelines for BE

Bioequivalence studies are

pharmacokinetic studies, they have
to follow the same GCP guidelines
as the clinical trials. The ICH-GCP
guidelines and Indian GCP
guidelines covers pharmacokinetic
studies under the clinical trial.
 Clinical trial
• 122-DAA
– “Clinical trial” means a systematic
study of new drug(s) in human subject(s)
to generate data for discovering and / or
verifying the clinical, pharmacological
(including pharmacodynamic and
pharmacokinetic) and /or adverse effects
with the objective of determining safety
and / or efficacy of the new drug.
 IMPORTANT CONSIDERATIONS

i. For drugs approved elsewhere in the world and

absorbed systemically, bioequivalence with the
reference formulation should be carried out.
ii. Evaluation of the effect of food
iii. Dissolution and bioavailability data to be
submitted
iv. All bioavailability and bioequivalence studies
should be conducted according to the
Guidelines for Bioavailability and
Bioequivalence studies as prescribed (ICMR
guidelines)