Biopharmaceutics and

Pharmacokinetics
Faculty of Pharmacy, University of Sadat city
Spring (2023-2024)
Course coordinator: Isra H. Ali, Ph.D.
Lecturer of Pharmaceutics
Prepared By: Hend Abdel-bar, Ph.D.
Associate Professor of Pharmaceutics
1
Biopharmaceutics

Objectives
• Bioavailability and
bioequivalence

2
Bioavailability and Bioequivalence
Absolute bioavailability compares the
bioavailability (estimated as area under
Bioavailability is a the curve, or AUC) of the active drug in
measurement of the extent of a systemic circulation following non-
therapeutically active drug that intravenous administration (i.e., after
oral, rectal, transdermal, subcutaneous
reaches the systemic administration), with the bioavailability
circulation and is available at of the same drug following intravenous
the site of action. administration.
𝐴𝑈𝐶 𝑝𝑜∗𝑑𝑜𝑠𝑒 𝐼𝑉
F= 𝐴𝑈𝐶 𝐼𝑉∗𝑑𝑜𝑠𝑒 𝑝𝑜

Relative bioavailability measures the

bioavailability (estimated as area under
the curve, or AUC) of a certain drug
when compared with another formulation
of the same drug, usually an established
standard, or through administration via a
different route
𝐴𝑈𝐶 𝐴∗𝑑𝑜𝑠𝑒 𝐵
F= 𝐴𝑈𝐶 𝐵∗𝑑𝑜𝑠𝑒 𝐴
3
Objectives of Bioavailability Studies

Primary stages of development of

a suitable dosage form for a new Determination of influence of Development of new formulations
drug entity to obtain evidence of factors affecting drug absorption of the existing drugs
its therapeutic utility

Control of quality of a drug

Comparison of availability of a
product during the early stages of
drug substance from different
marketing in order to determine
dosage forms or from the same
the influence of processing
dosage form produced by
factors, storage and stability on
different manufacturers
drug absorption

4
 Factors influencing bioavailability:
Factors related
Physicochemical factors: absorption: to Dosage form related
factors:

Dissolution and solubility- Noyes

Whitney equation, micronization, SAA pH partition theory and limitations Route
effects
Salt formation, Drug crystal forms
(amorphous, hydrates and solvates, physiological consideration Dosage forms
chirality)
Adsorption, complexation, micellar
solubilisation lymphatic absorption excipients effect

5
 Measurement of bioavailability
I. Pharmacokinetic methods
1) Plasma Level—Time Studies

Based on the Single Dose

assumptions:
Studies:
Pharmacokinetic Two dosage forms that exhibit
profile reflects the
therapeutic
superimposable plasma level-
time profile or urinary
Multiple Dose
excretion rate versus time result
effectiveness of a drug in identical therapeutic activity Studies:
6
 a) Single Dose studies:
At least 3 sample points should
I.V dose sampling should start
be taken if the drug follows
blood samples are collected for within 5 minutes of drug
one-compartment kinetics & 5
2 to 3 biological half-lives after administration and subsequent
to 6 points if it fits two-
drug administration. samples taken at 15 minute
compartment model to describe
intervals.
disposition process.

In Oral dose, at least 3 points The points for disposition or

should be taken on the descending phase of the curve Bioavailability is determined
ascending part of the curve for must be taken in a manner using Cmax, Tmax , AUC
accurate determination of Ka. similar to that for IV dose.

7
 b) Multiple Dose Studies:

A blood sample should be taken at

Drug administered for 5 biological Dosing interval should be greater
the end of previous dosing interval
half-lives (time to reach the than or equal to biological half-
and 8 to 10 samples after the
steady-state). life.
administration of next dose.

Bioavailability can also be

The extent of bioavailability is determined from the peak plasma
given as Fr and τ is the dosing concentration at steady-state Css,max
interval as follow as follow
[AUC]test Dstd τ test (Css ) Dstd
Fr= Fr= (Css max )test Dtest τ test
[AUC]std Dtest τstd max std τstd

8
2. Urinary Excretion Studies

It is based on the principle that the urinary excretion of unchanged

drug is directly proportional to the plasma concentration of drug

Bioavailability using urinary excretion data

This is widely used for drugs extensively
is valid if at least 20% of administered dose
excreted in urine or if urine as site of action.
is excreted unchanged in the urine

Estimation of bioavailability by urinary excretion

method shows high degree of variability & is less
reliable than plasma studies. But is used in conjunction
with blood level data for confirmatory purposes.

9
 2. Urinary Excretion Studies Method

Drug is administered to Collection of urine for Analysis of unchanged

the patient 7 biological half lives drug

(dXu/dt) max: Maximum urinary excretion Determination of

rate & analogous to Cmax its value↑ rate & amount of drug
extent of Abs ↑ excreted in each
interval and cumulative
(tu) max: Time for maximum urinary amount of drug
excretion rate. Analogous to Tmax. excreted
10
2. Urinary Excretion Studies Method
[Xu∞]oral D iv
• F=
The extent of [Xu∞]iv D oral
Xu∞ test D st
• F=
bioavailability is Xu∞ st D test
• Xu: Cumulative amount of drug excreted in the urine
given by

Bioavailability • Fr=
(Xu ss) test Dstd τ test
(Xu ss)std Dtest τstd
multiple dose study at • Xu,ss is the amount of drug excreted unchanged at
steady-state
steady state is given
by
11
Pharmacokinetic methods

Bioavailability can also be

determined for a few drugs
by assay of biologic fluids

theophylline- salivary
excretion

cephalosporin in CSF and

bile

12
II. Pharmacodynamics methods
Acute Pharmacological Response Disadvantages

Bioavailability is determined by
construction of pharmacological effect-
time curve as well as dose-response graphs.
The pharmacological response like Observed response may
The method requires ECG or EEG readings, pupil be due to an active
measurement of responses for diameter tends to be more variable metabolite whose
and accurate correlation between concentration is not
at least 3 biological half-lives. measured response and drug Conc. considered for the
is difficult. pharmacological effect.

It includes determination of
onset of action, duration of
action, intensity of action

13
III. Clinical trials

Limitations:
Improper quantification of observed
response.
based on observing the
clinical response to a drug Assumes that physiological status of the
patient does not change significantly over the
formulation given to patients duration of study.

suffering from disease for A patient who required the drug for disease
would be able to receive only single dose of
drug every week or for a few days.
which it is intended to be
Drug -drug interaction.
used

14
 In vitro tests
Dissolution test Approaches of IVIVC
In vitro-in vivo correlation (IVIVC)
These studies are done in vitro in artificial In vitro dissolution alone will be
gastric fluid, artificial intestinal fluid, insufficient to predict its therapeutic
artificial saliva, and artificial rectal fluid.
The amount of drug released from the
efficacy Establishing a linear
dosage form is measured over time to Correlation between in-vitro dissolution relationship between the in
determine if drug dissolution plays a role in & in-vivo bioavailability must be
limiting the absorption rate of the drug. established to predict its therapeutic vitro and the in vivo parameters
efficacy

Using data from previous in-

IVIVC is mathematical model that vivo studies to modify the in-
describes the relationship between an in- Objective of developing IVIVC is to vitro to develop IVIVC
vitro property (rate and extent of enable the dissolution test to serve as an
dissolution) of a dosage form and an in- alternate for in vivo bioavailability
vivo response (plasma drug concentration studies.
or amount of drug absorbed)

15
 In vitro studies

Applications of IVIVC Correlations

To ensure batch-to-batch consistency in therapeutic
efficacy of a drug product based in vitro test Based on pharmacological
Based on urinary excretion response: acute
data: dissolution parameter pharmacological effect
To develop a new dosage form with desired in-vivo are related to urinary drug such as LD50 is related to
performance parameters any of the dissolution
parameters.
Validating dissolution specifications &
development of bio- waiver guidelines Based on the plasma level
data : here linear
Statistical moments
To estimate the magnitude of the error in predicting relationship between
theory: relate MDT to
the in-vivo bioavailability results from in-vitro dissolution parameter and
MRT
dissolution data plasma level data are
established
16
IVIVC studies
In vitro dissolution parameter In vivo plasma data parameters
Time for specific amount of drug to dissolve (e.g. 50% AUC, Cmax
of the dose)
Amount dissolved at a specific time point Fraction absorbed, absorption rate constant
Ka
Mean dissolution time Mean residence time , mean absorption
time

17
IVIVC levels
Level A
Highest category correlation representing point- to- point relationship between in vitro and in
vivo parameters. In vitro dissolution and in vivo absorption rate curves are superimposable
In vitro dissolution curve serves as an alternate for in vivo testing and can accurately predict
its therapeutic efficacy

Level B
Not a point-to-point correlation utilizing principles of statistical moment analysis. Here mean
dissolution time is compared to either the mean residence time or in vivo dissolution time.
Cannot justify changes in manufacturing or modification in formula based on level B correlation.
In vitro data cannot be used for in-vivo quality control standards
Level C
• It is a single point correlation. Relates one
dissolution time point (T50%) to one PK parameter
such as AUC, Tmax , Cmax. Useful as guide in
formulation development or quality control

18
Types of dissolution apparatus

19
Drug permeability
To produce a biologic response, the
drug molecule must first cross a
biologic membrane. The biologic The interrelationship of the
membrane acts as a lipid barrier to dissociation constant, lipid solubility, Data obtained from the basic
most drugs and permits the and pH at the absorption site with the physicochemical studies, specifically,
absorption of lipid soluble substances absorption characteristics of various pKa, solubility, and dissolution rate,
by passive diffusion, while lipid- drugs are the basis of the pH partition provide an indication of absorption.
insoluble substances can diffuse theory.
across the barrier only with
considerable difficulty if at all.

The partition coefficient, P, is a

To enhance these data, a technique
measure of the differential solubility
using the everted intestinal sac may This method allows evaluation of
of a compound in two immiscible
be used to evaluate absorption both passive and active transport.
solvents. The most commonly used
characteristics of drug substances.
solvent system is octan-1-ol/water.

The partition coefficient is the

descriptor of lipophilicity for neutral
compounds, or where the compound
exists in a single form.

20
 Drug permeability
For ionisable solutes, the compound may
exist as a variety of different species in
each phase at any given pH. D, the
logP and logD both describe the same
distribution coefficient, is the appropriate
Partition Coefficient (P) = [Compound] physical property (lipophilicity) it is
descriptor for ionizable compounds since
octanol / [Compound] water critical that we understand the differences
it is a measure of the pH-dependant
between them and apply them accordingly.
differential solubility of all species in the
octanol/water system (typically used in the
logarithmic form logD).

LogP describes lipophilicity for neutral

LogD is the appropriate descriptor for compounds only, and while it can be a
lipophilicity of ionizable compounds very useful reference point for the
because it accounts for the pH dependence comparison of overall trends it should be
of a molecule in aqueous solution. applied with caution, especially when
working with ionizable compounds.

21
 Caco-2 cell line:
The Caco-2 cell line is a
Although in vivo studies yield The permeability of the
human colon adenocarcinoma Caco-2 cells can also be used
much definitive information cellular monolayer may vary
cell line that differentiates in to study interactions of drugs
about drug permeability in with the stage of cell growth
culture and resembles the with the transporter P-gp
humans, they are tedious and and the cultivation method
epithelial lining of the human discussed below.
costly to perform. used.
small intestine.

Permeability studies using the

Drug permeability using the In some cases, the drug Caco-2 cell line have been
Caco-2 cell line has been permeability may appear to be suggested as a method for
suggested as an in vitro low due to efflux of drugs via classifying the permeability of
method for passively membrane transporters such as a drug according to the
transported drugs. P-gp. Biopharmaceutics
Classification System (BCS).

22
The parallel artificial membrane
permeability assay (PAMPA)
Artificial lipid membranes are supported on a filter between two fluid compartments,
one of which contains the drug candidate.

The rate of appearance into the opposite compartment is then measured to determine the
permeability of the compound.

Several models and variations of this approach are available, and investigators should
pay attention particularly to the lipid composition of the artificial membranes as well as
other experimental details. Notably, the PAMPA can only predict simple diffusional
permeability, which does not involve uptake or efflux transporters.

23
In silico method: Lipinski rule of five and
applications.
Lipinski's rule states that, in general, an
Lipinski's rule of five is a rule orally active drug has no more than one
of thumb to evaluate the drug violation of the following criteria:
permeability and absorption.
However, the rule does not No more than 5 hydrogen bond donors
predict if a compound is (the total number of nitrogen–hydrogen
pharmacologically active. and oxygen–hydrogen bonds)
The rule is important to keep in mind during drug No more than 10 hydrogen
discovery when a pharmacologically active lead bond acceptors (all
structure is optimized step-wise to increase the nitrogen or oxygen atoms)
activity and selectivity of the compound as well
as to ensure drug-like physicochemical properties A molecular
are maintained as described by Lipinski's rule. mass less than
500 daltons
Candidate drugs that conform
to the rule of 5 tend to have An octanol-water
lower attrition rates during partition coefficient (log
clinical trials and hence have P) that does not exceed 5
an increased chance of
reaching the market.
24
Biopharmaceutical classification system (BCS):
Class Sol Per Abs. Pattern Examples Challenges in Drug Delivery
The two properties of biopharmaceutical
interest, that is, the properties that predict I High High Well Diltiazem CR forms need to limit drug
the ability of the drug candidate to move absorbed Propranol release or dissolution since
from dosage form to receptor, are water ol absorption of released drug is
solubility and membrane permeability. Metoprolo rapid.
l
II Low High Variable Nifedipine overcome solubility or
Naproxen dissolution problems

III High Low Variable Insulin ↑ permeability

Metformi
n
IV Low Low Poorly Taxol Improve both dissolution and
absorbed Furosemid permeability
e

25
BCS drug for immediate–release drug
products and IVIVC expectations
Class Sol Per IVIVC Predicting
Expectations IVIVC From
dissolution data
I High High If dissolution rate is Yes
slower than gastric
emptying rate.
II Low High In vitro dissolution Yes
rate = in-vivo
dissolution rate
III High Low Absorption is rate No
determining no
IVIVC from
dissolution data
IV Low Low Limited no IVIVC No
26
Bioequivalence Studies

Chemical Pharmaceutical Therapeutic

Equivalence:
Equivalence: Equivalence: Equivalence:

relative term that two or more drug products two or more drug
two or more drug
compares drug products are identical in strength, products that contain
products contain the
with respect to a specific quality, purity, content the same
same active ingredient
characteristic/function or a uniformity and therapeutically active
in the same amount
defined set of standards disintegration and ingredient eliciting
dissolution characteristics identical
but may differ in terms of pharmacological
excipients used effects and can
control the disease to
the same extent.
Bioequivalence: Drug in two or more identical dosage forms reaches systemic
circulation at the same relative rate and extent i.e. their plasma concentration-time
profiles will be identical without significant statistical differences
27
Bioequivalence Studies
Types of bioequivalence studies
Objectives
If a new product is
intended to be a substitute
for an approved medicinal
product as a
In vivo
pharmaceutical
equivalent/alternative

In vitro
The equivalence with this
product should be shown or
justified in order to ensure
clinical performance

28
I. In vivo (Studies are conducted by
Pharmacokinetic and Dynamic methods
for)
1. Oral immediate release products
with systemic action
Narrow therapeutic margin as digoxin and phenytoin

Pharmacokinetics complicated by absorption, nonlinear

kinetics, pre-systemic elimination

Unfavorable physiochemical properties, 2. Non-oral immediate

release products

Documented evidence for bioavailability problems

No relevant in vivo data available

3. Modified release products
with systemic action

29
 II. In vitro (Bio-wavers: In vitro studies, i.e., dissolution
studies can be used as alternate to in vivo bioequivalence under
certain circumstances as follow)
1. New drug product differs only in strength
of the active substance it contains, provided
all the following conditions hold:

The qualitative
Pharmacokinetics are
linear.
composition is the 2. The drug product has
same.
been slightly reformulated
or manufacturing method,
The ratio between
active substance and
Both products are slightly modified without
produced by the same
excipients is the same
or the ratio between the
manufacturer at the affecting bioavailability
same production site
excipients is the same
30
II. In vitro (Bio-wavers: In vitro studies, i.e., dissolution
studies can be used as alternate to in vivo bioequivalence under
An acceptable
certain circumstances as follow) IVIVC with
similar in vitro
3. The drug product meets all the dissolution rate
as approved
following requirements medicinal
product
It is in solution or solubilized The product is
form intended for topical
administration, oral
administration but
Active ingredient is in the same not intended to be
concentration as the approved absorbed.
drug product For parental solution; same active substance with same
concentration, same excipient, oral solution; same active
substance with same concentration, excipient not affecting GI
The product contains no transit or absorption, gas, otic / ophthalmic/topical solution;
excipients that effect absorption same active substance with same concentration, same excipient
and inhalational product/ nasal spray; administered with or w/o
of the drug same device as reference product; prepared as aqueous solution;
same active substance with same concentration, same excipient.
31
Bioequivalence study designs
The Crossover Design
The Parallel Design
A crossover design is a modified, randomized block
design in which each block receives more than one
formulation of a drug at different time periods.
A parallel design is a complete randomized
design in which each subject receives one
and only one formulation of a drug in a
random fashion. It is preferred if: For bioavailability and bioequivalence studies, the
crossover design is viewed favourably because of the
A-the drug is following advantages:
potentially toxic or
has a very long
elimination half-life
1. Each subject serves as his or her own control. It allows a
B-the population within subject comparison between formulations.
2. It removes the intersubjective variability from the comparison
of interest between formulations.
consists of very 3. With a proper randomization of subject to the sequence of
ill patients formulation administrations, it provides the best unbiased
estimates for the differences (or ratios) between formulations.
32
Washout period:
The washout period is defined as the rest
period between two treatment periods.
A suitable washout period should be
long enough to return any relevant
changes that influence bioavailability to
baseline (usually, at three times the
blood-plasma elimination half-life of the
active ingredient).

33