Part I:

Biopharmaceutics

Topic: 4
Bioavailability and Bioequivalence

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 Bioavailability
 Bioavailability is defined as the rate and extent (amount) of absorption of

unchanged drug from its dosage form.

 Bioavailability studies on drug can be required for a variety of reasons :

 Results from clinical studies indicate that different drug products

produce different therapeutics results.

 Results from bioavailability studies indicate that different products are

not bioequivalent.
 Drugs with a narrow therapeutic range.

 Low solubility and/or large dose and absorption is considerably less

than 100%
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 Bioavailability of a drug from its dosage form depends upon 3 major

factors :

1. Pharmaceutics factors related to physicochemical properties of the

drug and characteristics of the dosage form

2. Patient related factors

3. Route of administration
 The influence of route of administration on drug`s bioavailability is

generally in order parenteral > oral > rectal > topical.

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 Most drug are administered orally in such case, the dose available to

the patient called as bioavailable dose.

 Bioavailable dose is often less than the administered dose.

 The amount of drug that reaches the systemic circulation is called as

systemic availability or simply availability.

 The term bioavailable fraction F, refers to the fraction of

administered dose that enters the systemic circulation.

F ═ Bioavailable dose/ Administered dose

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 Objective of Bioavailability studies

Bioavailability studies are important in the –

1. Primary stages of development of a suitable dosage form for a new

drug entity to obtain evidence of its therapeutic utility.

2. Determination of influence of

- excipients,

- patient related factors,

- possible interaction with other drugs on the efficiency of

absorption.

3. Development of new formulations of the existing drugs.

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4. Control of quality of a drug product during the early stages of
marketing in order to determine the influence of processing factors,
storage and stability on drug absorption.

5. Comparison of availability of a drug substance from different dosage

forms or from the same dosage form produced by different
manufacturers.

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 Significance of bioavailability
 Drugs having low therapeutic index, e.g. cardiac glycosides,

quinidine, phenytoin etc. and

 Narrow margin of safety e.g. antiarrythmics, antidiabetics, adrenal

steroids, theophylline .
 Drugs whose peak levels are required for the effect of drugs, e.g.

phenytoin, phenobarbitone, primidone, sodium valporate, anti-

hypertensives, antidiabetics and antibiotics.
 Drugs that are absorbed by an active transport, e.g. amino acid

analogues, purine analogues etc.

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 In addition, any new formulation has to be tested for its

bioavailability profile.
 Drugs which are disintegrated in the alimentary canal and liver, or

those which under go first pass metabolism. e.g.chlorpromazine etc.

 Formulations that give sustained release of drug, formulations with

smaller disintegration time than dissolution rate.

 Drugs with steep dose response relationship i.e. drugs obeying zero

order kinetics / mixed order elimination kinetics ( e.g. warfarin ,

phenytoin, digoxin, aspirin at high doses, phenylbutazone)

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 Components of bioavailability
 Rate of absorption – The rapidity with which the drug is absorbed.

- Rapid onset : conditions like acute attack of asthma, intense acute

pain

- Slower onset : To prolong duration of action.

- To avoid adverse effects.

 Extent of absorption -chronic conditions like Epilepsy.

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 Bioavailability and bioequivalence
 Bioavailability- indicates a measurement of the rate and extent

(amount) of therapeutically active drug which reaches the general

circulation.
 Bioavailability can be divided in to

1. "Absolute" bioavailability (F)

2. "Relative" or “Comparative” bioavailability

1. "Absolute" bioavailability (F) - is the fraction of an administered

dose which actually reaches the systemic circulation, and ranges
from F = 0 (no drug absorption) to F =1 (complete drug absorption).

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2. "Relative" or “Comparative” bioavailability- refers to the
availability of a drug product as compared to another dosage form or
product of the same drug given in the same dose.
 The relative bioavailability of product A compared to product B, both
products containing the same dose of the same drug, is obtained by
comparing their respective AUCs.
 Where drug product B is the reference standard.

Relative Bioavailability =AUCA/AUCB

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 Route Bioavailability (%) Characteristics

Intravenous 100 (by definition) Most rapid onset

(IV)

Intramuscular 75 to ≤ 100 Large volumes often possible; may be

(IM) painful

Subcutaneous 75 to ≤ 100 Smaller volumes than IM; may be painful

(SC)

Oral (PO) 5 to < 100 Most convenient; first pass effects may be
significant

Rectal (PR) 30 to < 100 Less first-pass effects than oral

Inhalation 5 to < 100 Often very rapid onset

Transdermal 80 to ≤ 100 Usually very slow absorption; used for

lack of first-pass effects; prolonged duration
of action
 Bioequivalence
 Bioequivalence means the absence of a significant difference in the

rate and extent to which the active ingredient or active moiety in

pharmaceutical equivalents or pharmaceutical alternatives becomes
available at the site of drug action when administered at the same
molar dose under similar conditions in an appropriately designed study.

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Definitions
 Equivalence is more relative term that compares one drug product

with another or with a set of established standards. Equivalence may be

defined in several ways:
 Chemical equivalence indicates that two or more dosage forms

contain the labeled quantities of drug.

 Clinical equivalence occurs when the same drug from two or more

dosage forms gives identical in vivo effects as measured by a

pharmacological response or by control of a symptom or a disease.

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 Therapeutic equivalence implies that one structurally different

chemical can yield the same clinical result as another chemical.

 Brand-name drug is a drug marketed under a proprietary, trademark-

protected name.
 Generic drug is the same as a brand- name drug in dosage, safety,

strength, how it is taken, quality, performance, and intended use.

 Pharmaceutical equivalents drug products are considered to be

pharmaceutical equivalents if they contain the same active

ingredient(s), have the same dosage form and route of administration,
and are identical in strength or concentration.

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 Pharmaceutical alternatives these are drug products that contain the

same active moiety but contain different chemical forms such as esters
or salts of the active moiety or they may differ from the innovator’s
product in the dosage form or strength.
 Reference listed drug (RLD) a reference listed drug is an approved

drug product to which new generic versions are compared to show that
they are bioequivalent.

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Factors affecting bioavailability
 Before the therapeutic effect of an orally administered drug can be

realized, the drug must be absorbed.

 The systemic absorption of an orally administered drug in a solid

dosage form is comprised of three distinct steps:

disintegration of the drug product

dissolution of the drug in the fluids at the absorption site

transfer of drug molecule across the membrane lining the GIT into

the systemic circulation.

 Any factor that affects any of these three steps can alter the drug's

bioavailability and thereby its therapeutic effect.

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 The various factors that can influence the bioavailability of a drug can

be broadly classified as dosage form-related or patient-related.

A. Bioavailability factors related to the dosage form
 Physicochemical properties of the drug

Particle size

Crystalline structure

Degree of hydration of crystal and salt or ester form

 Formulation and manufacturing variables

Amount of disintegrant

Amount of lubricant

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Special coatings, nature of diluents and compression force
 B. Bioavailability factors related to the patient
 Physiologic factors

Variations in pH of GI fluids

Gastric emptying rate/time and intestinal motility

Perfusion of GI tract

Age, sex, weight

Disease states

Variations in absorption power along GI tract

Pre-systemic and first-pass metabolism

 Interactions with other substances- food, fluid volume and other drugs

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 Effect of Food on Drug Absorption

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 Drug interactions affecting absorption

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 Evaluation of the Bioavailability of a Drug
 Bioavailability testing is a means of predicting the clinical efficacy of a

drug.
 The estimation of the bioavailability of a drug in a given dosage form

is direct evidence of the efficiency with which a dosage form performs

its intended therapeutic function.
 The evaluation of a drug product bioavailability study involves the

consideration of various factors.

1. Drug
 The drug substance in each product must be the same and the only time

23 that this rule is relaxed is in the case of pro-drug administration.

2. Drug product
 Usually the comparison is made between two (or more) similar

products, containing exactly the same chemical substance.

 However, different dosage forms can be compared (when they contain

the same drug).

 For example we could compare an IM dosage form with an IV one.

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3. Subjects:
 A number of factor are of concern; health, age, weight, enzyme status,

number.

A. Health

 Healthy volunteers are often preferred by drug product evaluation

studies.
 Informed consent should be obtained from each volunteer and some
biochemical and medical examination will be used to confirm their
medical state.
 For some drugs there may be special disease states which may
cause the exclusion of some volunteers.
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B. Age

 Subjects between the ages of 18 to 35 years are preferred.

C. Enzyme status

 Smokers or subjects taking certain other drugs may have altered

kinetics for the drug of interest.

 This can be caused by alteration of enzyme activity or by drug-drug

interactions.
 These effects add complications to a study and an attempt is usually

made to minimize these factors.

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D. Weight

 The apparent volume of distribution is usually proportional to weight

in subjects of normal weight and height. However, in overweight (or

underweight) subjects the V in L/kg maybe somewhat different.

E. Number

The number of subjects included in the study should be sufficient to

see any real differences in bioavailability.

Usually 10 - 20 subjects are used in these studies.

In clinical studies where the end-point is some clinical response,

much larger numbers are required because of the greater variability

in clinical response.
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F. Assay
The same assay method should be used for all phases of the study.

It is not much use using one assay for product A samples and another

assay for product B samples.

Also the assay method should be sensitive and specific.

G. Design

Usually a complete cross-over design is used.

With this design each subject receives all products with a wash-out

period between each dose administration.

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H. Data analysis

 The rate of absorption can be characterized by the pka value and also

the time of peak concentration.

 The extent of drug absorption is characterized by the F value or the

peak concentration or total AUC values.

1. In vitro dissolution and bioavailability

 Trials for correlating physicochemical property and availability

 Reduce or sometimes eliminate the need for bioavailability tests

 Cost, time and safety

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A. Disintegration tests

Most pertinent in vitro parameter – must disintegrate before

significant dissolution or absorption can occur

Poor index of bioavailability

B. Dissolution test

It is the most sensitive in vitro parameter most likely to correlate

with bioavailability
Two official methods –basket and paddle method

Formulation factors (particle size, lubricant, coating) result in

product to product variation (b/n or with in batches of drug product).

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 Instrument variance and absence of a standard method

 Degree of agitation

 Size and shape of container

Composition of dissolution medium (pH, ionic strength, viscosity, δ)

To of dissolution medium

Volume of dissolution medium

Evaporation

Hydrodynamic (flow pattern)

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 Difference b/n in vitro and in vivo environment

In vitro in one or two standardized solvents

In vivo various PH, enzyme, surface tension, motility, presence of

other substance and absorption surface

In vivo environment is far more complex, variable, unpredictable

Proper selection of the in vitro and in vivo parameters to be

correlated is critical in achieving

e.g., single point type – dissolution

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2. In vivo methods

A. Demonstration of a clinically significant effect

B. Quantification of pharmacological effect (response vs conc.)

C. Measurement of drug conc. in blood or urine

 Normal healthy adult (18- 35 years old) male, normal heights and

weight
 Blood

key parameters to note are Auc, Cmax , t m ax (rate and extent )

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 Urine

cumulative amount of unchanged drug in urine

Useful for drugs not metabolized extensively

 At least 20% should be excreted unchanged

 Urine should be collected five times the half-life

 Less reliable than Cp analysis used in conjugation with Cp

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 BA and BE get increased attention due to increased generic drug

availability
 Generic substitution is only possible for PE and BE

 Generic and brand product should not statistically different in Cp

 BE & TE are →used interchangeably

 FDA consider two products to be therapeutic equivalents if

PE

BE

Compliance with compendia standard

Adequately labeled

35 Manufactured by GMP
 Drugs are categorized in to 3 depending on various risk potential for

inequivalence
i. High risk potential
Examples: Aminophiline, Digoxin, Warfarin
ii. Moderate risk potential
Examples: Ampiclin, Digitoxin , Griseofulvin
iii. Low or negligible risk potential

Examples: Acetaminophen, Codeine, Ferrous sulfate

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 Biopharmaceutics classification systems

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 Biopharmaceutics classification systems…

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 Biopharmaceutics classification systems…

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 Biopharmaceutics classification systems…

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 Biopharmaceutics classification systems…

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 Assessment of Biopharmaceutical properties

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 Assessment/Stability in GI fluids…

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 Assessment of Biopharmaceutical properties…

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 Assessment/Permeability…

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 Assessment/Partition coefficient…

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 Assessment/Partition coefficient…

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 Assessment/Partition coefficient…

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 Limitations of BA/BE studies
 Difficult for drugs with a long elimination half life.

 Highly variable drugs may require a far greater number of subjects

 Drugs that are administered by routes other than the oral route

drugs/dosage forms that are intended for local effects have minimal
systemic bioavailability.

E.g. ophthalmic, dermal, intranasal and inhalation drug products.

 Biotransformation of drugs make it difficult to evaluate the

bioequivalence of such drugs: e.g. stereoisomerism

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 Group Assignment

1. Distribution of drugs (G1 – Abdi Bedasa to Bontu Gadisa )

2. Protein binding of drugs (G2 – Chala Desalegn to Getwech Dech )

3. Biotransformation of drugs (G3 – Gelet Kebede to Lemi Tesfaye)

4. Prodrugs (G4 – Leta Adugna to Olira Dufera)

5. Excretion of drugs (G5 – Paulos Emiru to Gemechu Desta)

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1. Distribution of drugs (for G1)
 Introduction
 Tissue permeability of drugs
• Physicochemical prosperities of the drug
• Physiological barriers to distribution of drugs
 Organ/tissue size and perfusion rate
 Binding of drugs and perfusion rate
 Miscellaneous factors affecting drug distribution
 Volume of distribution

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2. Protein binding of drugs (for G2)
 Introduction
 Binding of drugs to blood components
• Plasma protein-drug binding
• Binding of drug to blood cells
 Tissue binding of drugs (tissue localization of drugs)
 Factor affecting protein-drug binding
• Drug related factors
• Protein/tissue related factors
• Drug interactions and patient related factors
 Significance of protein/tissue binding of drugs
 Kinetics of protein-drug binding
3. Biotransformation of drugs (for G3)
 Introduction
 Drug metabolizing organs and enzymes
 Chemical pathways of drug biotransformation
 Phase I reactions
• Oxidative reactions
• Reductive reactions
• Hydrolytic reactions
 Phase II reactions
• Conjugation with glucuronic acid
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• Conjugation with sulfate moieties
 • Conjugation with alpha-amino acids
• Conjugation with glutathione and mercapturic acid formation
• Acetylation
• Methylation
• Miscellaneous Conjugation reactions
 Factors affecting biotransformation of drugs
• Physiochemical properties of the drug
• Chemical factors
• Biological factors
 Bioactivation and tissue toxicity
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4. Prodrugs (for G4)
 Introduction
 Applications of prodrugs design
• Improvement of taste
• Change of physical form of the drug
• Reduction of GI irritation
• Reduction of pain on injection
• Enhancement of drug solubility and dissolution rate
• Enhancement of chemical stability of drug
• Enhancement of bioavailability (lipophilcity)
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 • Prevention of presystemic metabolism
• Prolongation of duration of action
• Reduction of toxicity
• Site-specific drug delivery (drug targeting )
 Limitation of prodrug design

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5. Excretion of drugs (for G5)
 Introduction
 Renal excretion of drugs
• Glomerular filtration
• Active tubular secretion
• Tubular reabsorption
• Concept of clearance
• Factors affecting renal excretion
• Dose adjustment in renal failure
• Dialysis and hemoperfusion
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 Non-renal routes of drug excretion
• Biliary excretion of drugs-enterohepatic cycling
• Plumonary excretion
• Salivery excretion
• Mammary and skin excretion
• Gastrointestinal and genital excretion

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