Bioavailability, Bioequivalence

and BCS system

By Dr. Ashwani Kumar Verma

10-07-2019 By Dr. Ashwani Kumar Verma 1

 Bioavailability and Bioequivalence
General concepts and overview

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 Drug: ADME

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Routes of Absorption, Distribution and Excretion
Routes of Absorption:
1. Ingestion
2. Inhalation
3. Dermal
4. Inhalation
5. Intravenous
6. Intraperitoneal
7. Intramuscular
8. Subcutaneous
Routes of Distribution:
1. Systemic circulation
2. Portal circulation
3. Lymphatic system
4. Fat
5. Extracellular fluid
6. Organs
Routes of Excretion:
1. Feces
2. Urine
3. Expired air
4. secretions
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 Bioavailability (F)
• The extent and rate at which its active moiety is delivered from pharmaceutical form and becomes
available in the systemic circulation
• Relative amount of an administered dose which reaches the general circulation and the rate at
which this occurs, i.e. Fraction of drug absorbed (F)
• Relative to either an IV dose (Absolute bioavailability) or
to another formulation of unknown F (Relative bioavailability).

Cmax Pharmacokinetics
conc. vs time
Conc.(mg/L)

Duration
of
Minimum action
effective
concentration

Onset
0.0
of 0 25

action Tmax Offset Time (h)

of
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action
 Pharmacokinetic measurement

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 Bioavailability and Factors Affecting It
1. The “true dose” is not the drug swallowed; BUT is the drug available to exert its effect.
2. Variable bioavailability may produce variable exposure
3. It is an Important component of overall efficacy of a drug

Drug’s
Pharmaceutial Patient’s related Clinical
Physicochemical
factors factors factors
properties
Physiologic factors

1. Chemical stability 1.Disintegration and dissolution time 1. Variations in pH of GI fluids Route of administration:
of drug 2.Pharmaceutical ingredients 2. Gastric emptying rate 1.Inhalation
3. Intestinal motility 2.Parenteral
2. Solubility (Water & 3.Special coatings 4. Pre-systemic and first-pass 3.Oral
lipid solubility) 4.Nature and type of dosage form metabolism/ Stability of drug to
enzymes 4.Topical/ Skin patches
3. Permeability 5.Rectal
5. Age, sex
• Particle size 6. Disease states/ Blood flow to GI 6.Vaginal
• Crystalline tract
structure 7. Specificity for carrier proteins and
• Salt form enzymes (Pgp or MDR1)
Interactions with other substances.
• Molecular size 1. Food
• pKa 2. Fluid volume
3. Other drugs
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 Bioequivalence
The term applied to generic formulations of the same active ingredient(API) and
dose, are said to be bioequivalent when the profiles of drug or its metabolite(s)
or both are similar (considers both rate & extent).

1. A comparison of the bioavailability of two or more drug products.

2. Two products or formulations containing the same active ingredient are

bioequivalent if their rates and extents of absorption are the same

3. Bioequivalence may be demonstrated through in-vivo or in-vitro test

methods, comparative clinical trials, or pharmacodynamic studies

Bioequivalence implies that with similar profiles, two bioequivalent drug products can be expected
to have the same systemic effect (both therapeutic and adverse)
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 Bioequivalence
90
80
Concentration (ng/mL)

70
60
Tes t/Generic
50
R eference/B rand
40
30
20
10
0
0 5 10 15 20 25 30
Time (hours)

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 NDA vs. ANDA Review Process
Original Drug Generic Drug
NDA Requirements ANDA Requirements
1. Chemistry
1. Chemistry
2. Manufacturing
2. Manufacturing
3. Controls
3. Controls 4. Labeling
4. Labeling 5. Testing
5. Testing 6. Animal Studies
6. Animal Studies 7. Clinical Studies
7. Clinical Studies 8. Bioequivalence
Study (In-vivo, In-
vitro)
Note: Generic drug applications are termed "abbreviated" because they are generally not required to include preclinical (animal) and clinical
(human) data to establish safety and effectiveness. Instead, generic applicants must scientifically demonstrate that their product is bioequivalent
(i.e., performs in the same manner as the original drug).
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 Bioequivalence: Factors
Pharmaceutical Equivalent
Products
Reference Test
Possible Differences
Drug particle size, ..
Excipients
Manufacturing process
Equipment
Site of manufacture
Batch size ….
Documented Bioequivalence
= Therapeutic Equivalence
(Note: Generally, same dissolution spec.)
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 FDA Definitions Used in Bioequivalence Determinations
Pharmaceutical equivalents Pharmaceutical alternatives Therapeutic equivalents
contain the same contain the same 1. Pharmaceutical equivalents
1. active ingredient(s), 1. therapeutic moiety, 2. Bioequivalent approved as
safe and effective
2. have the same dosage form 2. are different salts, esters, or
3. Adequately labeled
and route of administration, and complexes of the same moiety,
4. Manufactured in compliance
are identical in strength or are different dosage forms, or with current Good
concentration are different strengths Manufacturing Practice
regulations
may differ in characteristics, such Different dosage forms Therapeutic equivalents are
as and strengths within a expected to have the same
1. shape, single product line by a clinical effect and safety profile
2. release mechanisms, and single manufacturer
3. Packaging
Extended-release formulations
when compared with immediate-
or
standard-release formulations
Home-work: Collect 5 examples for each of the categories
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FDA Methods to Determine Bioequivalence
• Generic drug manufacturers must demonstrate that a
drug is bioequivalent to a reference drug product
• In order of FDA preference, methods used to define
bioequivalence
• Pharmacokinetic studies
• Pharmacodynamic studies
• Comparative clinical trials
• In-vitro/in-vivo studies
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 Pharmacokinetic Studies:
Key Measurements
Study Compound • AUC
Reference Compound • Area under the concentration- time curve
Cmax • Cmax
• Maximum concentration
• A difference of greater than 20% in Cmax or
Concentration

the AUC represents a significant difference

between the study and reference
compounds
• Tmax
• Time to maximum concentration
AUC

Tmax Time
 Conducting a Bioavailability Study-1
• Test and Reference given in a 1000
minimum number of subjects Test
(12) generally young males Reference

• Controlled Conditions on 100

Concentration (ng/mL)
separate occasions
• Assumption:
10
• Products shown to be
bioequivalent in young males
are assumed to be
1
bioequivalent in the population
that will use them, regardless
of age, sex, and ethnic
background. 0.1
0 2 4 6 8 10 12
Hours
 Conducting a Bioavailability Study-2
• Test and Reference in a 2-way 1000
cross-over design in a Test
minimum number of subjects Reference
(12) 100

Concentration (ng/mL)
• under controlled conditions

10
• Measure drug concentration
with a specific and sensitive
method
1

• Estimate AUC and CMAX

0.1
0 2 4 6 8 10 12
Hours
 Parameters for Estimating Bioavailability-1

• Estimating Rate: Is Cmax a good Cmax

measure of absorption rate?
7.0
• No ! ….
6.0
• Cmax is not a pure measure of rate …

Concentration (mg/L)
5.0
if we double the dose, Cmax will
4.0
change (ka might not) But it is easy to
3.0
estimate and in a profile there is no Tmax
argument about the highest 2.0

measured concentration. 1.0

0.0
0.0 2.0 4.0 6.0 8.0 10.0

Measures of Rate: Hours

Maximum concentration [Cmax],
absorption rate constant [ka]
 Parameters for Estimating Bioavailability-2

AUC
• Estimating Extent:- Is AUC a good 7.0
measure? 6.0

Concentration (mg/L)
• Yes ! …. 5.0

• Measures of Extent: Area Under 4.0

the Curve [AUC] Answer:

3.0

2.0

1.0

0.0
0.0 2.0 4.0 6.0 8.0 10.0

Hours
 Study Designs
1. Comparative Clinical Studies
Pharmacokinetic profile not possible
Lack of suitable pharmacodynamic endpoint e.g. Nasal
suspensions
• Single-dose, two-way crossover vs parallel design
• Multiple-dose studies
• Fast vs Fed
2. In-vivo (PK-PD)
3. In-vitro

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 Crossover vs. Parallel Designs
• Crossover design preferred
• Intra-subject comparison
• Lower variability
• Generally fewer subjects
required
The main problem with the cross
over design is: The carry over
effect………!
• Parallel design may be useful
• Drug with very long half-life
• Crossover design not practical

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 Multiple-dose and Fasted vs. Fed Designs
1. Multiple-dose design are employed in case of/when:-
• Drugs too potent/toxic
• Extended/modified release products
2. Fasted vs. Fed Design
Fasted study design preferred
• Minimize variability not attributable to formulation
• Better able to detect formulation differences
Fed Study Designs may be employed when:
• Significant gastrointestinal (GI) disturbance caused by fasted
administration
• Product labeling restricts administration to fed state

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 Comparative Pharmacodynamic (PD) Studies

• Not recommended when:

• active ingredient is well absorbed into the systemic
circulation
• pharmacokinetic study can be conducted and concluded
• Local action / no systemic absorption e.g. : Topical
Corticosteroid

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When equivalence studies are NOT necessary?
(Biowaivers)
• Aqueous parenteral solutions
• Solutions for oral use ( syrups, elixirs, tinctures & other
soluble forms but not suspensions)
• reconstitution as a solution
• ophthalmic aqueous solutions
• Topical aqueous solutions
• Nasal sprays or aqueous nebulizing inhalations

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 Waiver of Bioavailability and Bioequivalence
Studies for Immediate-Release Solid Oral
Dosage Forms Based on a
Biopharmaceutics Classification System
(B.C.S)

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 BCS Classifications
According to the BCS, drug substances are
classified as follows:

Class I - High Permeability, High Solubility

Class II - High Permeability, Low Solubility
Class III - Low Permeability, High Solubility
Class IV - Low Permeability, Low Solubility

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 Conditions for BCS Bio-waivers
Firms can request waivers of in vivo testing for Class 1 drug substances
Drug products must meet these criteria:
1. Immediate-release solid oral dosage forms

2. Highly soluble1, highly permeable2 drug substance

3. Rapid in vitro dissolution3

1. Solubility:- A drug substance is considered highly soluble when the highest dose strength is soluble in 250 ml
or less of aqueous media over the pH range of 1 - 7.5 (WHO , pH: 1.2 – 6.8)
2. Permeability:- A drug substance is considered to be highly permeable when the extent of absorption in
humans is determined to be 90% or more of an administered dose ( WHO, 85% ).
3. Dissolution:- A drug product is considered to be RAPIDLY DISSOLVING when > 85% of the labeled amount of drug
substance dissolves within 30 minutes using USP apparatus I or II in a volume of < 900 ml buffer solutions.

Note: Waivers not applicable for narrow therapeutic range therapeutic range (Digoxin,
Lithium, phenytoin, warfarin) drugs
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BCS-based biowaiver study: Requirements
Requirements for a BCS-based biowaiver study include:
1. Dissolution Test in 3 different media (in 900 ml and at 37°C) which are:
• Buffer pH 1.2, simulated gastric fluid (SGF) without enzymes or 0.1N
HCl.
• Buffer pH 4.5.
• Buffer pH 6.8 or simulated intestinal fluid without enzymes.
2. 12 samples in each media, paddle rotating at 50 rpm or basket at 100 rpm
3. Sampling times are 10, 15, 20, 30, 45 and 60 minutes.
4. The profiles of the test and reference products must be similar in all three
media.
The products are similar if the similarity factor f2 ≥ 50 and both products show
≥ 85% dissolution in 15 min.
USP Apparatus I (100 rpm) or II (50 rpm)

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 Summary
• Drug Bioavailability and Biopharmaceutical Factors Affecting it
• Pharmacokinetics(PK):- Basic consideration,
• Drug Product Performance, In Vivo: Bioavailability and Bioequivalence: Drug
Product Performance, Purpose of Bioavailability Studies, Relative and Absolute
Availability. Methods for Assessing Bioavailability,
• Bioequivalence Studies, Design and Evaluation of Bioequivalence Studies,
Study Designs, Crossover Study Designs, Generic Biologics (Biosimilar Drug
Products), Clinical Significance of Bioequivalence Studies.

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 Back-up slides

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 Biopharmaceutical
Consideration
1. Introduction,
2. Biopharmaceutical Factors Affecting Drug Bioavailability,
3. Solubility: Experimental methods.
4. Permeability: In-vitro, in-situ and In-vivo methods.
5. In-Vitro: Dissolution and Drug Release Testing, Alternative
Methods of Dissolution Testing Transport models,
6. Biopharmaceutics Classification System.

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 Pharmacokinetics(PK) and
Toxicokinetics(TK)
• Pharmacokinetics(PK):- Basic consideration
• Drug interaction (PK-PD interactions),
• The effect of protein-binding interactions, The effect of tissue-binding interactions,
• Cytochrome P450-based drug interactions,
• Drug interactions linked to transporters.
• Microsomal assays
• Toxicokinetics-Toxicokinetic evaluation in preclinical studies, Importance and applications of
toxicokinetic studies. LC-MS in bioactivity screening and proteomics.

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 Metabolite identification
• In-vitro / in-vivo approaches, protocols and sample preparation.
• Microsomal approaches (Rat liver microsomes (RLM) and Human liver microsomes (HLM) in
Met –ID. Regulatory perspectives. In-vitro assay of drug metabolites & drug metabolizing
enzymes.
• Drug Product Performance, In Vivo: Bioavailability and Bioequivalence: Drug Product
Performance, Purpose of Bioavailability Studies, Relative and Absolute Availability.
Methods for Assessing Bioavailability,
• Bioequivalence Studies, Design and Evaluation of Bioequivalence Studies, Study
Designs, Crossover Study Designs, Generic Biologics (Biosimilar Drug Products),
Clinical Significance of Bioequivalence Studies.

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 Rate versus Extent of Absorption

Extent of absorption is reflected by AUC

Rate of absorption, ka, is reflected by Tmax
Both Rate and Extent of absorption affect Cmax

Leads to 4 possible relative scenarios:

􀂄 (R) Rapid, (E) Complete Absorption•
yields a short Tmax, high Cmax, high AUC
􀂄 (R) Rapid, (E) incomplete absorption•
yields a short Tmax, low Cmax, low AUC
􀂄 (R) Slow, (E) complete absorption•
yields a long Tmax, high Cmax, high AUC
􀂄 (R) Slow, (E) incomplete absorption•
yields a long Tmax, low Cmax, low AUC
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