This In-house presentation was given as part of MSc. Clinical Research and consist only the Design and Conduct of BA/BE Studies of CDSCO Guideline. (INDIA)

1. Design and Conduct
of
BA/BE Studies
Central Drug Standard Control Organization’

2. Different Types of Studies :
 Pharmacokinetic Studies.
 Pharmacodynamic Studies.
 Comparative Clinical trials
 In-vitro studies.

3. Pharmacokinetic Studies.
1. Study design.
2. Study population.
3. Study condition.
4. Characteristics to be investigated.
5. Bioanalytical methodology.
6. Statistic evaluation
7. Sp. consideration for modified release drug product.

4. 1.Study design.
 ? To be answered.
 Nature of I.P.
 Evaluation method.
 Benefits/risk.
 Design
 period, phases, sequences & washout period.
 other designs.
 Single dose studies demand steady state for:
 Dose or time dependency, modified release products,
plasma concentration, intra-individual variability.

5. 2.Study population.
Selection of number of subjects
•Error variance with 1o characteristics
•Significance value should be 0.05.
•Expected deviations.
•Power
•Number of subjects (min 16).
•Sp. Cases se in subjects.
Selection criteria of subjects.
•To minimize the intra and inter subject variability.
•Risk to women
•For elderly subjects (60 years or more)
•Sex ratio.
•Narrowing the subjects.
• Risk of toxicity.
Genetic phenotyping.
•Considered for exploratory bioavailability for crossover and II studies.
•Genetic polymorphism.

6. 3.Steady State Studies.
Selection of Blood Sampling Points.
•Single dose-immediate release product -3 ½ lifes.
•Sampling Points-
•3 During Absorption Phase
•3-4 During Projected Tmax.
•4 During Elimination Phase.
•Interval Between two Sampling Points- not over ½ hour.
•Urine analysis.
Fasting and Fed State Considerations.
•Fasting State:
•Single Dose -Fasting (10+4) hours
•Multiple dose- Fasting (2 ) hours before and after dose.
• Fed State: Drug Under Fed State/modified release product.
•Fed state is maintain by High fat breakfast (950-1000 Kcal.)
15 min before dose.
Steady State Studies.
•Long Elimination ½ life and single dose blood concentration is not maintained
•Assay sensitivity.
•Compulsion of Cytotoxic Drugs.
•Modified release products to see dosage fluctuations in plasma.
•Drugs with intrinsic capabilities like their own metabolism.
•Enteric-coated preparations where coating is innovative.
•Combination products.
•Drugs with non-linear pharmacokinetics.
•Drugs likely to accumulate in body.

7. 4.Characteristics to be Investigated.
BA/BE evaluation other than
Biological Matrix.
• Concentration of drug/ metabolite is Low in Biological
matrix.
• Limitation of Analytical method.
• Unstable or stable Drugs / metabolites.
• Drugs with short half Life.
• In Case of Prodrugs.
• Intrinsic Characteristics of drug like spatial arrangement
i.e. racemate mixtures ( where achirality assay should be
performed).

8. 5.Bioanalytical Methodology
{A} Pre-Study Phase
Stability of Drug/ Metabolites in BM.
•Standard Condition for experiment.
•Freezing and Thawing data.
Specificity/Selectivity.
•Integrity of assay
Sensitivity.
•Capacity of test procedure.
Steady State Studies.
•Elimination ½ life and single concentration is not maintained
•Assay sensitivity.
Precision and Accuracy.
•Precision by replicate assay at several concentrations.
•Accuracy atleast 3 concentrations(low, medium, high).

9. 5.Bioanalytical Methodology
{A} Pre-Study Phase
Recovery.
•Documentation of concentration (High, medium, low).
•If low recovery, alternative method of Investigation.
Range and Linearity.
• Quantitative Relation between concentration and
response.
• Linear relation by standard curve with 5 concentrations
Analytical System Stability.
•Assay Precision should be monitored.
•Analytical standard at start and end of assays.

10. 5.Bioanalytical Methodology
{B} Study Phase
* Acceptable variability :
“Analysis of biological samples can
be done by single determination without need of
reanalysis.”
* Duplicate analysis as per case to case.
* Standard curve for each analyte & used for
unknown sample analyte concentration.
* No extrapolation of unknown sample.
* Instead re-determine Std. Curve or re-analyze
sample after dilution.
* Quality control sample accept/reject.

11. 5.Bioanalytical Methodology
{C} Quality Control Samples
* QCS:
“Samples with known concentration
prepared by spiking biological fluid with drug.”
* QCS and Standard Samples preparation at
different concentrations..
* QCS with stable analyte preparation in fluid of
interest.
* Less stable analyte preparation daily/weekly.
* QCS concentration assays with study
concentration and reference concentration to that
days calibration standard.
* Batch reanalysis if control sample are within
± 15 of expected concentration.

12. 6.Statistical Evaluation.
:Data Analysis:
* Primary concern in BE assessment is to limit consumer’s
risk and minimize manufacturer’s risk.
:Deviation from Study Plan:
* Method of analysis should be defined in the
protocol.
* Method of handling drop-outs and for handling
biologically implausible outlier should be
mentioned.
* Post hoc exclusion of outlier is not
recommended.
* Scientific explanation for volunteer exclusion is
needed

13. 6.Statistical Evaluation
:Statistical Analysis:
• SOP in protocol & BE should lead symmetry in two
formulation.
• It should account the source of variations.
• 90% confidence interval for population means or 5%
significance level for parameter under consideration.
• Logarithmic transformation for Cmax and AUC before
statistical analysis. But not recommended.
• Tmax analysis if clinically relevant, using non-parametric
methods.

14. 6.Statistical Evaluation
:Criteria for Bioequivalence:
• BE range 80-125% to obtain 90% confidence interval, which
is equivalent to rejection of two one sided t-test with null
hypothesis of non-equivalence at 5% significance.
• Permissible difference in bioavailability:
• Narrow therapeutic index.
• A serious dose related toxicity.
• A steep dose/effect curve.
• Non-linear pharmacokinetic within therapeutic range.
• Suprabioavailability:
• Need of reformulation with fresh BE study or
• Clinical data supporting the application.

15. 7.Sp.Considerations for Modified
Release Product.
General.
• MRP Include delayed , sustained, immediate + sustained
delayed + sustained, immediate + delayed.
• The Product Should:
• Act as Modified release & claim label.
• Preclude dose dumping effect.
• Provide therapeutic performance.
• Produce plasma level for proposed dosing interval at steady
state.
• .
Study Parameters
• BA should be obtained.
• Factor consideration: First market entry and Extent of
accumulation.
Study Design.
• Single and Single + Multiple dose depending on accumulation,
both in fasting and non-fasting state.
• Effect of food on reference:
• If known- two way crossover and
• If not known –three way crossover which include:
• --Reference in fasting state.
• --test in fasting state.
• --test in fed state

16. Pharmacodynamic Studies.
 Healthy or patients may be used.
 Necessary if quantitative analysis of drug (s) or their metabolites in plasma
or urine is not possible.
 Required if drug concentration is not surrogate endpoint for efficacy and
safety demonstration.
Requirements while Planning, Conducting and
Assessing Pharmacodynamic Studies.
• Response should claim efficacy and or safety.
• Method validation for precision, accuracy and specificity.
• Investigation of Dose response: should not be maximal of test & reference.
• Quantitative measurement of response under double blind condition and
recorded with instruments which act as substitute of plasma concentration.
• Non responder should be excluded prior to screening .
• Crossover or parallel design should be used.
• Statistical considerations for assessment.

17. Comparative Clinical Studies.
Items to be defined in the protocol in advance:
• Target parameters that represent the clinical
endpoints from which intensity
and onset of action can be derived.
• Size of acceptance range.
• Statistical method of analysis should be defined.
• Whenever applicable placebo design should be
defined.
• Include safety endpoints.

18. In Vitro-Studies.
Dissolution Studies:
• Carried out under mild condition at 370 ± 0.5 and
at physiological pH.
• More than 1 batch should be tested.
• Comparative dissolution profile rather than single.
• Design:
• Individual testing of 12 dosage units of each batch where
mean and individual results should be reported.
• % of nominal content released at spaced time to achieve
virtual complete dissolution.
• Dissolution profile in 3 aq media (1-6.8 or upto 8 pH).
• Same apparatus if possible on same or consecutive days.

19. By: Musale M.D. MSc. Clinical Research –Ist year.
“School of Biomedicalsciences, Aurangabad”
MGM’s University of Health Sciences,
Navi Mumbai.
E. Mail : mgmmusale@gmail.com