BIOPHARMACEUTICS 3

Expression of drug concentrations

Drug Bioavailability
Relative Bioavailability
Bioequivalence
Concentration and Dose -response curve
• One way to monitor the amount of drug in the body and its
effect at the site of action is to use a dose-response curve
• To measure analgesic response to a drug e.g. paracetamol, the
dose is increased and plotted against the degree of relief in
pain
• A specific dose is administered in a clinical trial to many
subjects and its effect measured
• A number of individuals respond to low doses, while some
respond to a higher dose to get an equal response. This is due
to Human variation
• It is therefore not ideal for relating the amount of drug in the
body to its effect
 Bioavailability
Definition: Refers to the rate and extent at which the active
moiety (drug or metabolite) enters circulation and thereby
accesses the site of action
• Rate at which drug becomes available to the body and the
extent to which the dose is ultimately absorbed after
administration
• It is the measure of the fraction of administered dose of a drug
that reaches systemic circulation in the unchanged form
 Bioavailability of drug given by IV bolus injection is 100%
 Bioavailability of drug after administration by a particular route
is calculated by comparing the plasma levels of the drug
relative to the level achieved by IV injection
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A typical blood plasma drug concentration-
time curve (oral admin)
https://1.bp.blogspot.com/-gb1DRJlFUKM/UmBMmZ5FLAI/AAAAAAAAAes/ktF967EFlOs/s1600/Screen+shot+2013-10- 17+at+4.45.57+PM.png

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 Blood plasma drug –time concentration curve
following IV and 3 oral dosage forms
https://www.boomer.org/c/p4/c20/Fig2012.jpg

• AUC

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Calculation of
Bioavailability
•Can be determined
by measurement of
drug concentration in
body fluids e.g. Whole
blood, plasma, serum,
urine or saliva.
•Calculated compared
to IV plasma levels
•Analytical method
sensitive to measure
parent compounds
and active metabolites

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 Absolute Bioavailability
• AUC= (F x Dose)/Total body clearance
• F=1 for IV bolus injection
• Foral = (AUC after oral administration)/
(AUC after IV bolus injection)
• F= Absolute Bioavailability of the drug
from the dosage form when compared to
IV administration of the same drug (as
the reference)

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 Relative Bioavailability
• F<1 via routes of administration other than IV
bolus
• F= Relative Bioavailability when the reference is
another dosage form (e.g. a solution form or
innovator brand) other than IV bolus injection
• Bioavailability variation of drugs with a low
therapeutic index (like digoxin) and where dosage
needs precise control are monitored closely given
that slight increases in fraction of absorbed dose
may lead to toxic effects
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 Drug Concentration in Urine
• Measurement of drug/metabolite concentrations in urine
can be uses to assess bioavailability
• Advantages:
– is safer, easier and less intrusive and traumatic for the subject
than blood sampling
– Urine has less proteins
– Has higher drug concentrations thus simpler to analyze
– Ideal for drugs and metabolites extensively excreted in urine
– Rate of urinary excretion and cumulative amount excreted can be
calculated
– Where the rate of drug excretion is proportional to the plasma
concentration, the urinary excretion rate – time curve will be
similar in shape to the plasma concentration-time curve (there
are exceptions to this proportionality)
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 Drug Concentration in Urine ctd.
• Absolute Bioavailability (F) =
( Uoral /UIV) x 100
(Uoral and UIV are cumulative amounts excreted
after oral and /IV administration respectively.

• Disadvantage: incomplete collection of urine

from subjects

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 Saliva concentration of drug
• Drugs excreted in saliva in good concentration
(e.g. paracetamol, digoxin, phenytoin, litium, etc.)
• It has been proven that concentrations in saliva
are proportional to blood plasma concentrations
• Measurement of salivary excretion rate provides
an alternative method to assess bioavailability
• Adv: sample collection is simple, non-invasive,
• Caution: orally administered drug could
contaminate early sample-concentrations if
residue is left on tongue
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 Bioequivalence 1
Drug formulations are deemed bioequivalent if:
The rate and extent of absorption of drug from the
test products do not show a significant difference
from the rate and extent of absorption of drug
from the reference product when administered at
the same molar dose of the therapeutic agent
under similar experimental conditions in either a
single dose or multiple doses
OR

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 Bioequivalence 2
The rate and extent of absorption of drug from the test
product do not show a significant difference from the
rate and extent of absorption of drug from the
reference product when administered at the same
molar dose of the therapeutic agent under similar
experimental conditions in either a single dose or
multiple doses, and the difference from the reference
product in the rate of absorption of the drug is
intentional , is reflected on the proposed labeling, is
not essential to the attaining of effective body drug
concentrations on chronic use , and is considered
medically insignificant for the drug.
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 Bioequivalence and Bioinequivalence
RELEVANCE
1. Prior to introduction of a new formulation to clinical
use, a demonstration of satisfactory bioavailability is
required by regulatory authorities. When:
– Original manufacturer wishes to vary excipients or change
the process of manufacture
– A product similar to one already on the market is being
introduced
– A product not identical but contains same active entity as
existing licensed one is being introduced, e.g. a salt,
isomer, hydrate, etc. form,
– Its a new product with modified release claims
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 Relevance
2. When inequivalence would be hazardous to the patient,
e.g.:
– Change of product could lead to increase in bioavailability
for drug with narrow therapeutic index, toxicity concerns
– In case of inefficacy of dose patient could suffer serious
consequences of the disease, e.g. anti-epileptic s,
anticoagulants, etc
– Product is intended for high risk patient groups; e.g.
pediatric or geriatric patients
– Drug product is a modified release dosage form thus
contains more drug than a normal single dose, risk of dose-
dumping
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 When bioequivalence tests are not required

• Bioequivalence studies are not required when:

– It has been demonstrated that an in vitro
dissolution test is an accurate predictor of
performance in vivo
– IV and topical preparations do not require
bioequivalence testing
– (accumulated information on the therapeutic
agent, dosage form, formulation and disease state
should be considered to justify testing).

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 Clinical equivalence
– used to ensure that products from different sources or
manufacturers can be used interchangeably.
– requires demonstration in clinical practice that the
same therapeutic effect is produced as measured by
control of disease or symptomatic relief
– It is possible for products that are not bioequivalent to
be clinically equivalent
– Drugs with a narrow therapeutic index that require
careful control of plasma concentration, bioequivalence
is paramount, the reference product being of proven
clinical equivalence
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 Bioequivalence Tests
• Are performed in vivo in clinical trials
• In healthy volunteers
• Each product administered as a single dose under
fasting, in random order in a cross-over design
• Study must have significant statistical power to
detect variances (Sample size)
• Rate of formation of an active metabolite after
administration should be assessed as part of the
bioequivalence study
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 In vitro – in vivo correlation
• Laboratory (dissolution) tests are not
predictive of release rates from a formulation
in vivo
• However, experience has shown that
formulations with poor release characteristics
in vitro often present (predict) low
bioavailability in clinical study, and vice versa.

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Factors affecting the bioavailability of a drug
1. First-pass hepatic metabolism
2. Solubility of the drug
3. Chemical stability
4. Drug formulation factors:
– Affecting absorption from the GIT,
• Use of inactive polymorphs
• Particle release and dissolution from dosage form
• Change in method of production
• Particle size variation for sparingly soluble drug
• brand variations

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Modification/Alteration of release from oral dosage
form:

• Formulation as a water-miscible mixture

• Reduction of particle size, increase area for dissolution
• Use of a more soluble salt
• A water soluble pro-drug
• Selection of a more soluble metastable form of drug (if
polymorphic)
• Selection of a solvated form (if non-solvated, solvated
forms)
• Molecular dispersion in a water soluble carrier, e.g.
cyclo-dextrins
• HW: When is a reduced release rate desirable?
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 Formulation factors affecting the
Absorption of drug
HW: look at factors affecting release:
• from injections
• From implants

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BIOPHARMACEUTICS 4

PHARMACOKINETICS
Drug Half-life
Simple Examples in PK
 PHARMACOKINETICS
Is the study of the quantitative
changes of absorption,
distribution, metabolism and
excretion of a drug with time,
following its administration to
man or animals.

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 PK

• Is the change of drug

concentration as it moves
through the different
compartments of the body

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 Concentration
• [Concentration]= Mass/Volume
• Mass=Amount of drug (mg or mcg)
• Volume (mL or L)
– Blood volume = 5L
– Plasma Volume = Blood – cells = 2.5L

Q1: If a 20mg dose of drug is 100%

absorbed, what is its plasma drug
concentration?
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 Solution 1
• Conc = Mass/Volume
Plasma drug concentration
= 20mg/2.5L
= 8mg/L

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 Drug Half-life t1/2
• Is the time it takes for the total amount of
drug in blood plasma to decline to one-half
an initial value, once the drug reaches peak
concentration
• It is the time it takes to eliminate 50% of the
drug at any given time
• It is often referred to as the Elimination Half-
life of the drug.

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Dosing intervals and drug removal
• Time to peak (tp)= is the time it takes for drug
to get to peak concentration (Cmax) after
administration of the dose
• Dosing intervals relate to half-life: in general,
the next dose of drug is given once drug
reaches its half-life
• Five times the half-life is used to estimated the
time it essentially takes to remove the drug
from the body
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 Example Q1
• Suppose after a single oral dose of drug P,
assessment of a plasma sample concentration
at time t=5hrs is 60mcg/mL, and at t=11hrs is
30mcg/mL;
SOLUTION 1
• the estimated half-life of the drug is:
11hrs - 5hrs =6hrs
• time it essentially takes to remove the drug
from the body
i.e. 5 x6hrs = 30hrs for drug P.
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 Example Q2
After administration of a single oral dose of
Drug AKs, the peak time is 11/2hours, and the
peak concentration is 1200mcg/mL. Its half
life is three hours. What will the plasma levels
be after 101/2 hours? Tabulate.

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 Solution 2
Time (hrs) Half -life Plasma concentrations
(mcg/mL)

1.5 hrs 0 1200

4.5 hrs 1st 600

7.5 hrs 2nd 300

101/2 hrs 3rd 150

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 Practical Significance of Drug Half-life

1. In general, the next dose is given

when the previous dose reaches its
half life.
• The normal dose interval is based on
one half life.
2. To know how long it takes to
eliminate all the drug from the body
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 Calculation of amount of drug remaining

• No matter the concentration of drug you begin with in plasma, 95% of

it will be eliminated after about 4 1/2 Half lives
• Half-life is inversely proportional to the rate of drug metabolism:
t1/2= 0.693/Ke ; where Ke is the first order elimination rate constant (i.e.
for most dosages, the proportion of drug metabolized over time is
constant)
• If the rate of elimination of drug(K e) increases, the half-life decreases
t1/2= 0.693/Ke =(0.693 Vd)/CL; where CL is the volume of from which drug is
completely removed per unit time (mL/min); V d
• Amount of drug remaining can be expressed as”
– A Fraction of peak concentration
– As a %
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 Amount of drug remaining
Time Plasma Drug Remaining
(Hrs) Concn Fraction %
(mcg/mL)

1.5 hrs 1200 1 100

4.5 hrs 600 ½ (1/2)1 50

7.5 hrs 300 ¼ (1/2)2 25

101/2 hrs 150 1

/8 (1/2)3 12.5
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 Why drugs have different Half-lives

Differences in:
• Structural
• Physicochemical properties

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 Drug examples: Aspirin and Naproxen
sodium
• https://www.sigmaaldrich.com/content/dam/sigma-aldrich/structure1/180/mfcd00002430.eps/_jcr_content/renditions/mfcd00002430-mediu
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• https://www.sigmaaldrich.com/content/dam/sigma-aldrich/structure9/178/mfcd00058507.eps/_jcr_content/renditions/mfcd00058507-large.
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Multiple dosing regimen
http://www.thebody.com/content/art875.html

Biopharmaceutics 38
 Multiple dosing regimen
• Is the manner in which the drug is administered in suitable
doses by a suitable route with sufficient frequency that
ensures maintenance of plasma concentrations within the
therapeutic window for the entire period of therapy.
• Dosing frequency: Is the time interval between doses
• Dose size: Is the quantity of drug administered [the larger
the dose size, the greater the fluctuation between
maximum concentration at steady state (Cssmax) and
minimum concentration at steady state (Cssmin]
• Both dose size and dose frequency can be adjusted.
• What is a trough concentration (CT)?
• What is concentration at steady state? Cssmax? Cssmin?
Biopharmaceutics 39
40 PK
Best of wishes!