SUBJECT: DOSAGE FORM DESIGN

SUBJECT CODE: BPHM 801

SEMESTER: 8TH
TOPIC: BIOAVAILABILITY AND BIOEQUIVALENCE
STUDIES
SUBJECT IN-CHARGE: MRS. PRABHJOT KAUR
Bioavailability and
Bioequivalence studies
Bioavailability
Indicates a measurement of the rate & extent of therapeutically active drug
that reaches the systemic circulation.
Equivalence
It is the relation in terms of bioavailability, therapeutic response, or a set
of established standards of one drug to another.

Generic substitution
Process of dispensing a different brand or unbranded drug product in
place of the prescribed drug product

Substituted drug product contains the same active ingredient or

therapeutic moiety as the same salt or ester in the same dosage form but
is made by the different manufacturer.
Pharmaceutical alternatives
Drug products that contain the same therapeutic moiety but as different salts,
esters, and complexes.
e.g. Tetracycline phosphate or hydrochloride equivalent to 250 mg Tetracycline
base
e.g. extended release & immediate release

Pharmaceutical equivalents
Drug products that contain the same active ingredient (same salt, ester, or
chemical form) & are identical in strength/conc., dosage form and route of
administration
Also chemical equivalents
Must meet identical standards (strength, quality, purity and identity) but
may differ in characteristics (e.g. color, flavor, shape, packaging,
preservatives)
 Pharmaceutic substitution
Process of dispensing a pharmaceutical alternative for the prescribed drug
product
Requires physician’s approval
e.g.. Amoxicillin capsules instead Ampicillin suspension
Therapeutic equivalents
These drugs are pharmaceutically equivalent and can be expected to
have the same clinical effect and safety profile.

Therapeutic alternatives
Drug products containing different active ingredients that are indicated for the
same therapeutic or clinical objective.
Therapeutic drug products may differ in certain characteristics (e.g. color,
flavor, packaging).
Purpose of Bioavailability Studies

Ensure the drug product is safe & effective for its labeled indication for use
prior to its marketing
Those requiring bioavailability studies
Unmarketed drugs which do not have full NDA (new drug application)

New formulations of active drug ingredients or therapeutic moieties that

have full NDA approval & are approved for marketing.
Drug with new NDA
Example: Metformin, antidiabetic drug given as treatment for Polycystic Ovarian
syndrome (PCOS).
Characterize the essential pharmacokinetics of the active drug ingredient (e.g. rate &
extent of systemic absorption, elimination half-life, rates of excretion and metabolism
after single or multiple dose administration)

New Formulation of a Drug that have full NDA approval

Example: Enalapril sustained release or sublingual tab in place of immediate release

tab,
 Paracetamol capsule in place of paracetamol IV

To determine the bioavailability and characterize the pharmacokinetics of the new
formulation, new dosage form or new salt or ester relative for a reference formulation
 Bioequivalence studies
 Relative and absolute Bioavailability

Absolute Bioavailability

 requires I.V. administration

 Ratio of the oral: intravenous AUC values normalized for
dose
 Fabs= (AUC oral / AUC *(Dose iv / Dose oral)
iv)

Relative Bioavailability

 no I.V. reference
 comparison AUC values (ratio) of different dosage forms /
formulations
 Frel = (AUC a / AUC b) *(Dose b /Dose a)
 5 5

4 4

Solution
Capsule
Conc. [mg/L]

Conc. [mg/L]
3 3

2
2

1
1

0
0
0 4 8 12 16 20 24
0 4 8 12 16 20 24
Time [hours]
Time [hours]

20 mg administered as 80 mg given as a solution

an i.v. bolus (Diovan) and a capsule (Diovan)
 F=0.6

100

Solution F=0.4*
Capsule 16
80
14
% of f remaining to be absorbed

F=0.2*
12
60
10

AUC
40 8
6
20 4
2
0
0
0 2 4 6 8 10
I.v. (20 mg) P.O. (80 mg) P.O. (80 mg)
Time [hours] Capsule Solution

*dose - adjusted
Methods of Assessing Bioavailability:

 1. PLASMA DATA
a. tmax - time of peak plasma concentration
- the time required to reach the maximum drug
concentration after drug administration.
- as rate of absorption === tmax

b. Cmax – maximum plasma drug concentration obtained

after the administration of the drug.
- indicates that the drug sufficiently and
systematically absorbed to provide a therapeutic response .
As,
- Dose === Cmax
-Absorption rate ==== Cmax

-
c. AUC
 Area Under the Curve

 Area under the plasma level time curve

 Measure of quantity of drug in the body

◦ If one knows the AUC for a given dose size upon IV administration of a
drug, the fraction of drug absorbed (F = absolute bioavailability) upon
EV route of administration can be calculated from the AUC obtained by
the latter route
It is directly proportional to the dose
Measures the extent of bioavailability
Unit: µg/ml.hrs or µg hrs/ml
Indepent of the route of administration and processes of drug elimination as
long as the elimination processes do not change
 Relative magnitude of the AUC can be obtained by either of the
following methods:
1. counting method
2. weighing method

Exact calculation of the AUC is done by either one of the following

two methods:
1. trapezoidal rule
2. using blood level equations
Trapezoidal rule: compute for the individual area of the trapezoids.
Total AUC = 0 → infinity

Infinite time – time beyond which the area is insignificant

 [AUC]0∞ = [AUC]0t + [AUC]t∞

 AuC1 = ( C0 + C1) ∙ ( t1 - t0)
2
Informations that we can get from a typical conc-time curve

 MEC – min. conc. required by the receptors to produce the desired

effect

 MTC – min. conc. needed to just barely produce toxic effect

 Desired therapeutic range (therapeutic window) – distance between

MEC and MTC; it is where the drug produces its characteristic effects

 Onset time – time required for drug to reach MEC

 Duration of action – the difference between onset time and the time for
the drug to decline back to MEC; length of time the drug level remains
above MEC
 2. urine data

Urinary excretion of the unchanged drug vs time data can be used to

Estimate the bioavailability of a drug product.
Acc. To this method the rate of urinary excretion of a drug is directly
proportional to the concentration of the drug in the blood

a. Du – cumulative amount of drug excreted in the urine

- it signifies, how much drug is present in the urine
- directly related to the total amount of drug absorbed.

Informations required to prepare/determine the patient’s urine profile

1. subjects & dose of drug
2. determine the total amount of urine collected
3. sample is analyzed for drug content
4. construct a graph of cumulative amount of drug in the urine vs time
b. rate of drug excretion in the urine dDu
dt
dDu/dt - dependent on the first order kinetics and the plasma
concentration .

c. time for the maximum urinary excretion (t∞) -

F= [XU∞]oral ₓ [Dose]iv/ [XU ∞] X [Dose]oral

Cumulative amount of drug in urine VS Time
Rate of drug excretion (dDu/dt) vs Time
3.Acute pharmacological effect

1. An established dose-related response curve.

2. An easily measurable pharmacological response such as heart rate,
ECG, blood pressure, pupil diameter, etc.

4. clinical response
- toxicity
- no response
- therapeutic failure
- good therapeutic
response
 Bioequivalence is a comparison of bioavailability of two or
more product i.e. Formulation containing the same active
ingredient are said to be bio-equivalent if their rate & extent
of absorption is same.
 One of the best way to compare two product is by clinical
studies. But performing clinical studies is extremely dificult
& expensive. Hence Bio-equivalence studies are generally
used to compare two products so as to compare their
biological equivalence.
 Bioequivalence is receiving increasing attention because of
increased availability & importance of generic drugs in
today's p’ceutical market.

 However bioequivalence is not straight forward for all the drugs .
Many drugs shows bioinequivalence.
 In 1973 ad hoc committee on drug product selection of American
Pharmaceutical Association published a list of drug that show
bioinequivalence.
 Based on this list drug has been divided into 3 categories
HIGH RISK POTENTIAL MODERATE RISK LOW RISK POTENTIAL
POTENTIAL
Aminophylline Amphetamine Acetaminophen
Bishydroxy coumarine Ampicillin Codeine
Digoxin Chloramphenicol Hydrochlorothiazide
phenytoin Digitoxin Ephedrine
prednisolone Erythromycin Isoniazide
prednisone Griesofulvin Meprobamate
quinidine Penicillin G Penicillin V
warfarin Pentobarbital Sulfixazole
 Bioequivalence Studies

 Difference in predicted clinical response may be due to difference

in pharmacokinetics, pharmacodynamics or bioavailability of drug
products .
 Bioequivalent drug products that have same systemic drug
bioavailability will have same predictable response .
 Difference in pharmacodynamics behavior is due to age, drug
tolerance, drug interaction etc.
 Bioavaliability of drug may be more during empty stomach
 When drug is used on daily basis, nature of an individuals diet &
lifestyle may affect plasma drug level because variable absorption
in the presence of food.
Bases for determining bioequivalence

 Bioequivalence is established if the in-vivo bioavaliability

of test drug product (usually generic product) does not
differ significantly in rate & extent of drug absorption,
from reference listed drug (brand name product)
administered at same dose, under similar experimental
conditions.
 BIOEQUIVALENCE PROBLEMS
Bioequivalence problem occurs due to following reason-
 Active drug ingredient has low solubility in water . (less than 5 mg/ml) .
 Dissolution rate is low.
 Certain structural forms of active drug ingredient (e.g. polymorphic
forms, solvates, complexes & crystal modifications) dissolve poorly,
thus altering the absorption.
 Drug product that have high ratio of excipients to active ingredients
(e.g. greater than 5:1) .
 Specific ingredients such as hydrophilic & hydrophobic excipient &
lubricant may interfere with absorption .
 Active ingredients absorbed in particular segment of GIT.
 The degree of absorption of active drug ingredient is poor when
administered in pure form.
 Rapid metabolism in intestinal wall or in liver during absorption
process.
BIOEQUIVALENCE STUDY

Parameters
1. AUC:- Area under plasma drug concentration time
curve. AUC Provides information regarding amount
of drug in plasma i.e. extent of release.

2. C max:- Maximum plasma drug concentration. Partly

depends on rate of drug release from the formulation.

3. T max:- Time required to reach maximum plasma

drug concentration
Tmax is also dependent on rate of drug release from
the formulations.
 Pharmacokinetic Studies
Key Measurements

Study Compound  AUC

Reference Compound  Area under the concentration- time
Cmax curve
 Cmax
 Maximum concentration
 A difference of greater than 20% in
Concentration

Cmax or the AUC represents a

significant difference between the
study and reference compounds
 Tmax
 Time to maximum concentration
AUC

Tmax Time
4. Plasma Through fluctuation(%):-

 This parameter is generally used in

bioequivalence studies of sustained release
formulation.

It can be calculated as
 %PTF= C max-C min
C average
Where C average is Avg Plasma drug conc. During
dosing period.
5. T1/2:- Elimination half life .It provides information
regarding elimination of drug from the body.
6. Normalized C max:- C max & T max are known
to show significant intersubject variability & hence
normalized C max is used in some cases.
It can be calculated as-
Normalized C max= C max
AUC
7. Mean residence time(MRT) : It provides an
idea about time a drug molecule spend in the body
before it get excreted.
1.Title
Study Protocol :-
a) Principle investigator( Study director)
b) Project/protocol number & date.
2.Study objective
3. Study design
a) Design
b) Drug products
1.Test products
2. Reference Product
c) Dosage regimen
d) Sample collection schedule
e) Housing/ confinement
f) Fasting/meal schedule
g) Analytical methods
4. Study population
a) Subjects
b) Subject selection
1. Medical history
2. Physical examination.
3. Laboratory test.
 c) Inclusion and exclusion criteria
d) Restriction / prohibitions
5. Clinical procedures
A) Dosage and drug administration
B) Biological sampling schedule
C) Activity of subject
6. Ethical Consideration
A) Basic principles
B) Institutional review board
C) Informed consent
D) Adverse reactions
7. Facilities
8. Data analysis
A) Analytical validation procedure
B) Statistical treatment of data
9. Drug accountability
10. Appendix
 STUDY DESIGN
 Bioequivalence study should be designed in such away so as to
identify treatment effects that are produced after administration of
products.
Two types of design
1. Crossover
2. Parallel
 Generally crossover is used in practice in this design both test and
reference product are compared in each subject that is each subject
act as his/her own control
 Hence inter subject variable such as age ,weight,
metabolism etc. are minimal.
 Because of minimal intersubject variability, crossover design is
widely used for routine studies.
 In this design first subjects are randomly divided in to 2 groups and
sequence of drug is randomly assigned.
ISF COLLEGE OF PHARMACY
 If there are two formulations – test (T) and
reference (R) ; in period 1 , group 1 will receive T
followed by R.
 Time period between administrations of two
formulations will be sufficiently long considering
adequate ‘wash out period’.
 Wash out period is essential to ensure complete
elimination of drug before next administration.
 10 half lives ensures more than 99% of elimination
and hence wash out period should be at least 10
half lives.
 Period II will start after completion of wash out period now in period II,
group 1 will receive ‘R’ followed by ‘T’ .
 If drug require washout period of 7 days then sequence of administration
of 2 formulation can be shown as

Group Period I (day1) Period II (8 day )

I T R

II R T

 Thus sequence of administration for group I is T-R & group II is R-T .

 Two formulation trial is always a 2 period trial,
 Thus in period I , 50% subject ( group I) receive T & 50% subject
(group II) receive R .
 In period II order is reversed
Latin square cross over
design
 For biequvalence study in human volenteers
 Compares 3 different drug formulations- A,B,& C.
 In this design , each subject receives each drug
product only once
 Adequate time is provided between medications
for elimination of drug from the body.
 In this design each subject is having his /her own
control hence subject to subject variation is
reduced.
LATIN –SQUARE CROSS OVER DESIGN FOR BIOEQUIVALENCE STUDY OF 3 DRUG
PRODUCTS IN 6 HUMAN VOLUNTEERS

SUBJECT STUDY PERIOD 1 STUDY PERIOD 2 STUDY PERIOD 3

1 A B C

2 B C A

3 C A B

4 A C B

5 C B A

6 B A C
 Subject:-
 This study is usually performed with healthy volunteers of
both sexes.
 Usually subjects should be between 18-55 yrs old.
 Weight of subject is within normal range.
 It is important to reduce the intra & inter subject variability
 Products are usually administered after overnight fasting &
meal taken should standardized.
 During the study subject should not taking any other
medication .
 Subject should not allowed to take any alcoholic or xanthine
containing beverages e.g. coffee, tea ,cola ,soft drinks etc.
 If subject is smoker then this should be mentioned in their
medical history.
 Reference & Test Product:-
 In bioequivalence study the test product is usually compared
with reference product.
 Reference product is generally the innovator product (Product
of company who 1st invented this product.)
 Before proceeding study the both the test product & reference
product are tested for in vitro dissolution profile.
 Evaluation of data:-
Analytical Method:-
 The analytical method for measurement of drug
must be validated for accuracy, precision ,sensitivity,
& specificity.
 Only 1 analytical method is used during
bioequivalence study because different method may
yield different values.
 Data should be presented in both graphical &
tabulated form for evaluation.
Pharmacokinetic evaluation of data
 For single dose studies-
Pharmacokinetic analysis include calculation of
AUC, Cmax, Tmax.
 For multiple dose-
Calculation of steady state AUC, Tmax, Cmax,
Normalized Cmax.
Statistical evaluation of data-
This can be done by using analysis of variance
(ANOVA) test.
REFERENCES

Venkateswarlu V. , “Biopharmaceutics and Pharmacokinetics”, edition 2004,

PharmaMed press, Hyderabad.

Brahmankar D.M., Jaiswal B. Sunil, “bipharmaceutics and pharmacokinetics- a

treatise”, first edition 1995, Vallabh prakashan, New Delhi.
Sharge leon , yu pc andrew, applied and pharmacokinetics, third edition.
Thank you