BE studies- Part 3 : Statistical Phase,

Special Situations, Guidelines

Dr. Ammar Raza

Clinician, Clinical Affairs
Manager, Medical
Advisor
Introduction
 Performance will never be identical
– Two formulations
– Two batches of the same formulation?
– Two tablets within a batch?
 Purpose of bioequivalence (BE)
– Demonstrate that performance is not
“significantly” different
– Same therapeutic effect
– What constitutes a ‘significant’
difference?
Phases of BE studies
Focus on

 Clinical Phase  Bioanalytical  Statistical Phase

– Screening Phase – Data analysis
– Selection – Storage of – Anova (SAS)
Samples – PK (WinNonlin)-
– ICF &
– Method Devpt AUC, Cmax etc.
Recruitment
– Method Validation – T/R ratio
– Dosing
– Analysis of – 90% CI
– Sampling samples
– Monitoring – QC checks
– AEs
BE Studies: Scientific Basis

Two different formulations of a drug

resulting in SIMILAR systemic
concentration-time profiles will always
achieve similar concentration time
profiles at the site of efficacy or toxicity
Metrics for BE studies
 Concentration vs. time profiles
– Area under the curve (AUC)
Observable exposure AUC-t (zero to last detectable concn) Overall

 Complete exposure AUCinf extent
– Maximal concentration (Cmax) Both rate
– Time to Cmax (Tmax) & extent
 Statistical measures of BE metrics
– Mean
– Variance
 Stat BE: comparing the means of two products
PK analysis: Approaches

 BEor rate & extent of BA or exposure can be

proven using
– Compartmental approach - not preferred in BE
– Non-compartmental approach
 Based on calculation of AUC –body’s exposure
 Favored to prove BE- robustness
 Min of 15 samples- to calculate AUC
Non-compartmental approach
Determination of BE

 Modern concept of BE is based on a survey of

physicians carried out by Westlake in 70s - a 20%
diff in dose b/n 2 formulations - no clinical
significance for most drugs
– BE limits were set at 80 - 120%.
 Pl concn. dependent measures - Cmax or AUC are
not normally distributed
– Are log normal,
– BE limits became 80 - 125% (or ± 0.225 on natural log
scale)
Statistical Determination of BE

 Past method
– tested null hypothesis- no difference between means
 Not adequate in 1980s
 Current method
– proves similarity between two products
– BE- diff <20%
 Avg rate & extent of BA of T within ±20% that of R
– Log transformed scale- limits of ratio b/n 0.8 and 1.25
The statistical procedure ..
 ‘two one-sided test’
– introduced by Hauck
 Method – defines error α- probability of concluding
BE when in reality it is not true
– Usually fixed to a min- 5%
– BE concluded if 90% CI of the ratio is within 80-125
– Power of study (regulatory)=80%
– 20% probability of not demonstrating BE even if they are
truly BE
– No of sub: based on variability of metric that study must
pass on
 Cmax: most variable metrics
Sample size
Analysis of Variance

 ANOVA
– Most common technique of analysis and estimation
 Lognormal distribution
– Raw data must be log transformed
– Comparison of means & variances of transformed data
– Geometric mean (GM)
– Results reported in original scale
Confidence Intervals (CI)

 Inference from study to wider world

 Range of values within which we can have a
chosen confidence that the population value
will be found
 Study findings expressed in scale of original
data measurement
Confidence Intervals (cont.)
 Width of CI indication of (im) precision
of sample estimates
 Width partially dependent on:
– Sample size
– Variability of characteristic being
measured
 Between subjects
 Within subjects
 Measurement error
 Other error
Confidence Intervals cont.
 Degree of confidence required
– More confidence = wider interval
 Width of equivalence limits represents allowable
boundary for ratio (or difference) of means b/n
products in comparison
 In other words, width of CI dependent on:
– Standard error (SE)
 Standard deviation, sample size
– Degree of confidence required
Statistical Analysis
(Two One-sided Tests Procedure)

 Statistical analysis of pharmacokinetic

measures
– Confidence intervals
– Two one-sided tests
 AUC and Cmax
– 90% Confidence Intervals (CI) must fit between
80%-125%
 Typical BE
Assessment Criteria

 90% confidence interval

 Ratio of geometric means
 Acceptance criteria: 80 – 125%

 Log transformed AUCT & Cmax

Statistical Approaches for BE
 Average bioequivalence
 Population bioequivalence
 Individual bioequivalence

Average BE
 Conventional method
 Compares only population averages
 Does not compare products variances
 Does not assess subject x formulation
interaction
Statistical Approaches for BE

 Population and individual BE

– Include comparisons of means and variances
 Population BE
– Assesses total variability of the measure in the
population
 Individual BE
– Assesses within subject variability
– Assesses subject x formulation interaction
Statistical effects in model

 Sequence effect
 Subject (SEQ) effect
 Formulation effect
 Period effect
 Carryover effect
 Residual
Statistical Analysis
 Bioequivalence criteria
– Two one-sided tests procedure
 Test(T) is not significantly less than reference
 Reference (R) is not significantly less than test

 Significant difference is 20% (α = 0.05 significance

level)
– T/R = 80/100 = 80%
– R/T = 80% (all data expressed as T/R so this
becomes 100/80 = 125%)
Special Situations
 Highlyvariable drugs
 Endogenous substances
 Parent/ metabolite issues
 Long half-life drugs
Highly Variable Drugs (HVDs)

 Intrasubject variability (CV%) ≥30%

– Significant first pass metab or to a poor or erratic
absorption process
 Sample size in BE studies is determined -by
BA parameter with highest variability
– most cases, Cmax has higher variability than
AUC
 May not pass even when the reference
product is tested against itself
Factors Contributing to the Variability
 Related to Formulation  NON-related to
– Disintegration Formulation
– Dissolution  Absorption:
– Permeability – Rate of GI transit:
Stomach to the colon
– Transport through GI mucosa
 Pancreatic or bile acid
secretion
 Drug metabolism
– induction
– inhibition
– Liver blood flow
 Excretion
– Renal blood flow
HVDP

 highly variable drug product (HVDP) - formulation of

poor pharmaceutical quality - drug itself is not highly
variable- big component of within formulation
variability (WFV)
– cannot be detected in traditional 2-treatment, 2-period, 2-
sequence cross-over design studies
– Replicate designs- facilitate their detection - within-subject
variabilities of test & reference formulations can be
estimated separately
 When they are v. different - one of the formulations is a HVDP
Approaches
 Evaluate bioequivalence at steady-state
– Variability expected at steady-state is < that after single dose
– Always true?
– Can not be applied to all HVD/HVDP
 Assessment of BE on the metabolite
– When parent undetectable- metabolite is less variable
– smaller sample size - BE studies for HVD if based on the
metabolite
 Add-on
 Individual BE- may help overcome existing problems ABE
 Average BE with scaling approach & widen CI
IBE Vs. ABE

 Study periods are duplicated 4 vs. 2

– Bad: duration & cost x 2
– Good: may reduce pool size of volunteers - HVD
 Hasnot found unanimous consensus in the
scientific community
– Remains under investigation
– Subject of discussion in future
Wider CI

 Major regulatory agencies have provisions -

can accommodate effect of higher variability
associated with Cmax on design of BE
studies
 EMEA -expanded limits (e.g., 75-133%) for
Cmax in certain cases -NO safety or efficacy
concerns
 MCC, SA - allow for expanded limits for
Cmax in certain cases
Example

 2 BE studies on formulations of drugs A & B

– same no. of sub in each study
– GMR is the same in both
– Two One-Sided Test - only difference b/n 2 studies is
magnitude of CV
 Drug A - low within-subject variability (ANOVACV
15%) - 90% CI falls comfortably within BE limits
 Drug B is highly variable - ANOVA-CV of 35%
– Study on drug B was underpowered- simple remedy -
repeat study with a greater no. of sub
Progesterone
The Poster Drug for High Variability

A repeat measures study of Prometrium®

2x200 mg caps in 12 healthy PM females
yielded:
Intrasubject CV for AUC of 61%
Intrasubject CV for Cmax of 98%
 Generic company calculated that a 2 period
crossover BE study - require dosing in 300 PM
women to achieve adequate statistical power
Endogenous substances

 Pose a major problem

 Baseline levels present
– Administration of drug can alter levels / feedback
mechanism
– Oral admin- frequently produces only a negligible
inc in baseline; wide variability
– Baseline should be measured throughout the day
before dosing
Endogenous substances

Issues
 100s or 1000s of vols to operate with net post-dose
values
– Not acceptable from ethical or financial point of view
 Lesser no of vols- post dose values without baseline
subtraction
 Steady state studies
– preferred design when possible
 Assay sensitivity issues
Predominating active metabolite

Parent/ metabolite issues

 Parent more variable
 Difficult to get detectable concn. (absent or
marginally present)
 Rate of abs- adequately evaluated only assaying
parent
 Measure metabolite
– Less of subs reqd
– Easier to prove BE
Allopurinol, flutamide, terfenadine
Long half-life drugs
 Crossover- adequate washout to avoid carryover –
study lasts 4-6 m or more
 Parallel design
– More no. of sub (n=18 in crossover is stat equivalent to
~n=50 in parallel)
 Costly
 Approaches: steady state or truncated AUC
(stopping at 24 or 48 h)
– crossover -washout cannot be shortened, duration of study
partially reduced
– Parallel design- markedly reduce duration of study
Topical application

Three classes
 Administered topically for absorption into sys
circulation e.g. patches
– can use usual BE
 Designed to exert topical activity only – absorption is
negligible e.g. ointments, creams
– PD study or clinical efficacy study
 Designed to exert local activity – absorbed to a
certain extent only e.g. vaginal prep etc.
– Considered individually
BE vs. Clinical Trial: Differences

Clinical Trial BE study

 Multicentric  Single centre
 Subjects: mostly  Subjects: Mostly
patients (except Ph I) healthy vol; rarely pts.
 Multiple doses  Single dose; sometimes
 Costly and time multiple dose
consuming  Cheaper and require
less time
Focus on FDA Guidance

 Two main guidances

– General : “Bioavailability and Bioequivalence Studies for Orally
Administered Drug Products — General Considerations”
http://www.fda.gov/cder/guidance/5356fnl.pdf
– “Food effect bioavailability studies and fed Bioequivalence studies”
http://www.fda.gov/cder/guidance/5194fnl.pdf
– Drug specific guidances
 Levothyroxine
 Potassium hydrochloride
– Biowaiver
– Retention samples
Hatch-Waxman Amendments
to FFD&C Act - 1984

 Considered one of the most successful pieces

of legislation ever passed
 Created the generic drug industry
 Increased availability of generics
 1984 12% prescriptions were generic
 2000 44% prescriptions were generic - yet only 8% of
revenue for prescription drugs
 Compromise legislation to benefit both brand
and generic firms
Hatch-Waxman Amendments
to FFD&C Act - 1984

 Allowed generic firms to rely on findings of

safety and efficacy of innovator drug after
expiration of patents and exclusivities (do not
have to repeat expensive clinical and pre-
clinical trials)
 Allowed patent extensions and exclusivities to
innovator firms
Requirements for generic drugs

• Labeling
• Chemistry/Microbiology
• Bioequivalence
• Legal
Labeling

• “Same” as brand name labeling

• May delete portions of labeling protected by
patent or exclusivity
• May differ in excipients, PK data and how
supplied
Chemistry

• Components and composition

• Manufacturing and controls
• Batch formulation and records
• Description of facilities
• Specs and tests
• Packaging
• Stability
Manufacturing Compliance
Programs
 Purpose - To assure quality of marketed
drug products
 Mechanisms - Product Testing
– Surveillance
– Manufacturing/Testing plant inspections
– Assess firm’s compliance with good
manufacturing processes
Guidance for CROs

Scope: Guidance to organizations involved

in the conduct and analysis of in vivo
bioequivalence (BE) studies

Note: BE studies should be performed in

compliance with:

• General regulatory requirements

• Good clinical practice (GCP)
• Good laboratory practices (GLP)
 Guidelines

Guideline provides information on:

- organization and management
- study protocols
- clinical phase of a study
- bio-analytical phase of a study
- pharmacokinetic & statistical analysis
- study report
Comparison of guidelines
Importance

 Understandingthe generic drug approval

process and the issues surrounding BE is of
paramount importance to both clinicians and
scientists

Welage LS, Kirking DM, Ascione FJ, Gaither CA.J Am Pharm Assoc (Wash). 2001 Nov-
Dec;41(6):856-67
Resources

Text book of pharmacokinetics

http://pharmacy.creighton.edu/pha443/pdf/pkin08.pd
Summary
 Planning is important
– Study design, sample size, sampling schedule, incl & excl criteria
 Conduct
– Clinical & ethical: Protocol approval, selection of volunteers,
housing, dosing, sampling, AE recording and reporting, ambulatory
samples,
– Bioanalytical
 assay method, equipment (HPLC / LC MS/MS), SOPs
– PK & Statistical
 Software (WinNonlin, SAS etc.)
 Reporting
– 3 Parts, CRFs, TMF, Chromatograms
 Thank You
dr.razadiacare@rediffmail.com
Biowaiver

 Recommended for a solid oral Test product that

exhibit rapid (85% in 30 min) and similar in vitro
dissolution under specified conditions to an approved
Reference product when the following conditions are
satisfied:
– Products are pharmaceutical equivalent
– Drug substance is highly soluble and highly permeable
and is not considered have a narrow therapeutic range
– Excipients used are not likely to effect drug absorption