be studies- part 3 : statistical phase,

special situations, guidelines

dr. ammar raza
clinician, clinical affairs manager, medical advisor
introduction
l performance will never be identical
l two formulations
l two batches of the same formulation?
l two tablets within a batch?
l purpose of bioequivalence (be)
l demonstrate that performance is not “significantly” different
l same therapeutic effect
l what constitutes a ‘significant’ difference?
phases of be studies
l clinical phase
l screening
l selection
l icf & recruitment
l dosing
l sampling
l monitoring
l 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
l concentration vs. time profiles
l area under the curve (auc)
l observable exposure auc-t (zero to last detectable concn)
l complete exposure aucinf
l maximal concentration (cmax)
l time to cmax (tmax)
l statistical measures of be metrics
l mean
l variance
l stat be: comparing the means of two products
pk analysis: approaches
l be or rate & extent of ba or exposure can be proven using
l compartmental approach - not preferred in be
l non-compartmental approach
l based on calculation of auc –body’s exposure
l favored to prove be- robustness
l min of 15 samples- to calculate auc
non-compartmental approach
determination of be
l 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
l be limits were set at 80 - 120%.
l pl concn. dependent measures - cmax or auc are not normally distributed
l are log normal,
l be limits became 80 - 125% (or ± 0.225 on natural log scale)

statistical determination of be
l past method
l tested null hypothesis- no difference between means
l not adequate in 1980s
l current method
l proves similarity between two products
l be- diff <20%
l avg rate & extent of ba of t within ±20% that of r
l log transformed scale- limits of ratio b/n 0.8 and 1.25

the statistical procedure ..

l ‘two one-sided test’
l introduced by hauck
l method – defines error α- probability of concluding be when in reality it is not true
l usually fixed to a min- 5%
l be concluded if 90% ci of the ratio is within 80-125
l power of study (regulatory)=80%
l 20% probability of not demonstrating be even if they are truly be
l no of sub: based on variability of metric that study must pass on
l cmax: most variable metrics

sample size
analysis of variance
l anova
l most common technique of analysis and estimation
l lognormal distribution
l raw data must be log transformed
l comparison of means & variances of transformed data
l geometric mean (gm)
l results reported in original scale
confidence intervals (ci)
l inference from study to wider world
l range of values within which we can have a chosen confidence that the
population value will be found
l study findings expressed in scale of original data measurement
confidence intervals (cont.)
l width of ci indication of (im) precision of sample estimates
l width partially dependent on:
l sample size
l variability of characteristic being measured
l between subjects
 l within subjects
l measurement error
l other error
confidence intervals cont.
l degree of confidence required
l more confidence = wider interval
l width of equivalence limits represents allowable boundary for ratio (or difference) of
means b/n products in comparison
l in other words, width of ci dependent on:
l standard error (se)
l standard deviation, sample size
l degree of confidence required
statistical analysis
(two one-sided tests procedure)
l statistical analysis of pharmacokinetic measures
l confidence intervals
l two one-sided tests
l auc and cmax
l 90% confidence intervals (ci) must fit between 80%-125%
typical be
assessment criteria
l 90% confidence interval
l ratio of geometric means
l acceptance criteria: 80 – 125%
l log transformed auct & cmax
statistical approaches for be
l average bioequivalence
l population bioequivalence
l individual bioequivalence
statistical approaches for be
l population and individual be
l include comparisons of means and variances
l population be
l assesses total variability of the measure in the population
l individual be
l assesses within subject variability
l assesses subject x formulation interaction
statistical effects in model
l sequence effect
l subject (seq) effect
l formulation effect
l period effect
l carryover effect
 l residual
statistical analysis
l bioequivalence criteria
l two one-sided tests procedure
l test (t) is not significantly less than reference
l reference (r) is not significantly less than test
l significant difference is 20% (α = 0.05 significance level)
l t/r = 80/100 = 80%
l r/t = 80% (all data expressed as t/r so this becomes 100/80 = 125%)

special situations
l highly variable drugs
l endogenous substances
l parent/ metabolite issues
l long half-life drugs

highly variable drugs (hvds)

l intrasubject variability (cv%) ≥30%
l significant first pass metab or to a poor or erratic absorption process
l sample size in be studies is determined -by ba parameter with highest
variability
l most cases, cmax has higher variability than auc
l may not pass even when the reference product is tested against
itself
factors contributing to the variability
l related to formulation
l disintegration
l dissolution
l permeability

l non-related to formulation
l absorption:
l rate of gi transit:
stomach to the colon
l transport through gi mucosa
l pancreatic or bile acid secretion
l drug metabolism
l induction
l inhibition
l liver blood flow
l excretion
l renal blood flow

hvdp
l highly variable drug product (hvdp) - formulation of poor pharmaceutical quality - drug
itself is not highly variable- big component of within formulation variability (wfv)
l cannot be detected in traditional 2-treatment, 2-period, 2-sequence cross-over design studies
l replicate designs- facilitate their detection - within-subject variabilities of test & reference
formulations can be estimated separately
l when they are v. different - one of the formulations is a hvdp
approaches
l evaluate bioequivalence at steady-state
l variability expected at steady-state is < that after single dose
l always true?
l can not be applied to all hvd/hvdp
l assessment of be on the metabolite
l when parent undetectable- metabolite is less variable
l smaller sample size - be studies for hvd if based on the metabolite
l add-on
l individual be- may help overcome existing problems abe
l average be with scaling approach & widen ci
ibe vs. abe
l study periods are duplicated 4 vs. 2
l bad: duration & cost x 2
l good: may reduce pool size of volunteers - hvd
l has not found unanimous consensus in the scientific community
l remains under investigation
l subject of discussion in future

wider ci
l major regulatory agencies have provisions - can accommodate effect of
higher variability associated with cmax on design of be studies
l emea -expanded limits (e.g., 75-133%) for cmax in certain cases -no
safety or efficacy concerns
l mcc, sa - allow for expanded limits for cmax in certain cases
example
l 2 be studies on formulations of drugs a & b
l same no. of sub in each study
l gmr is the same in both
l two one-sided test - only difference b/n 2 studies is magnitude of cv
l drug a - low within-subject variability (anovacv 15%) - 90% ci falls comfortably within
be limits
l drug b is highly variable - anova-cv of 35%
l study on drug b was underpowered- simple remedy - repeat study with a greater no. of sub
progesterone
the poster drug for high variability
endogenous substances
l pose a major problem
l baseline levels present
l administration of drug can alter levels / feedback mechanism
l oral admin- frequently produces only a negligible inc in baseline; wide variability
l baseline should be measured throughout the day before dosing
endogenous substances
issues
l 100s or 1000s of vols to operate with net post-dose values
l not acceptable from ethical or financial point of view
 l lesser no of vols- post dose values without baseline subtraction
l steady state studies
l preferred design when possible
l assay sensitivity issues
predominating active metabolite
parent/ metabolite issues
l parent more variable
l difficult to get detectable concn. (absent or marginally present)
l rate of abs- adequately evaluated only assaying parent
l measure metabolite
l less of subs reqd
l easier to prove be
long half-life drugs
l crossover- adequate washout to avoid carryover – study lasts 4-6 m or more
l parallel design
l more no. of sub (n=18 in crossover is stat equivalent to ~n=50 in parallel)
l costly
l approaches: steady state or truncated auc (stopping at 24 or 48 h)
l crossover -washout cannot be shortened, duration of study partially reduced
l parallel design- markedly reduce duration of study
topical application
three classes
l administered topically for absorption into sys circulation e.g. patches
l can use usual be
l designed to exert topical activity only – absorption is negligible e.g. ointments, creams
l pd study or clinical efficacy study
l designed to exert local activity – absorbed to a certain extent only e.g. vaginal prep
etc.
l considered individually
be vs. clinical trial: differences
clinical trial
l multicentric
l subjects: mostly patients (except ph i)
l multiple doses
l costly and time consuming

be study
l single centre
l subjects: mostly healthy vol; rarely pts.
l single dose; sometimes multiple dose
l cheaper and require less time

focus on fda guidance

l two main guidances
l general : “bioavailability and bioequivalence studies for orally administered drug products — general
considerations” http://www.fda.gov/cder/guidance/5356fnl.pdf
l “food effect bioavailability studies and fed bioequivalence studies”
http://www.fda.gov/cder/guidance/5194fnl.pdf
l drug specific guidances
 l levothyroxine
l potassium hydrochloride
l biowaiver
l retention samples

hatch-waxman amendments
to ffd&c act - 1984
l considered one of the most successful pieces of legislation ever passed
l created the generic drug industry
l increased availability of generics
l 1984 12% prescriptions were generic
l 2000 44% prescriptions were generic - yet only 8% of revenue for prescription drugs
l compromise legislation to benefit both brand and generic firms
hatch-waxman amendments
to ffd&c act - 1984
l 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)
l allowed patent extensions and exclusivities to innovator firms

requirements for generic drugs

l labeling
l chemistry/microbiology
l bioequivalence
l legal
labeling
l “same” as brand name labeling
l may delete portions of labeling protected by patent or exclusivity
l may differ in excipients, pk data and how supplied
chemistry
l components and composition
l manufacturing and controls
l batch formulation and records
l description of facilities
l specs and tests
l packaging
l stability
manufacturing compliance programs
l purpose - to assure quality of marketed drug products
l mechanisms - product testing
l surveillance
l manufacturing/testing plant inspections
l assess firm’s compliance with good manufacturing processes
guidance for cros
guidelines
comparison of guidelines
importance
l understanding the generic drug approval process and the issues
surrounding be is of paramount importance to both clinicians and
scientists

resources
text book of pharmacokinetics
http://pharmacy.creighton.edu/pha443/pdf/pkin08.pdf

summary
l planning is important
l study design, sample size, sampling schedule, incl & excl criteria
l conduct
l clinical & ethical: protocol approval, selection of volunteers, housing, dosing, sampling, ae recording and
reporting, ambulatory samples,
l bioanalytical
l assay method, equipment (hplc / lc ms/ms), sops
l pk & statistical
l software (winnonlin, sas etc.)
l reporting
l 3 parts, crfs, tmf, chromatograms

thank you
dr.razadiacare@rediffmail.com
biowaiver
l 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