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:Use of in vitro studies

Full biowaiver

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Bioequivalence
Pharmaceutical Equivalent
Products
Reference Test
Possible Differences
Drug particle size, ..
Excipients
Manufacturing process
Equipment
Site of manufacture
Batch size ….

Documented Bioequivalence
= Therapeutic Equivalence
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Full biowaiver for new formulation (e.g. generic)

No in vivo bioequivalence study necessary

The in vivo BE study is waived based upon:

 Solubility (drug property)
 permeability
 comparability in excipients
 comparative dissolution data
Concept: Biowaivers based on
Biopharmaceutics Classification System
(BCS)
high permeability low permeability

high HS/HP HS/LP

solubility Class I Class III

LS/HP LS/LP
low
Class II Class IV
solubility
 BCS VIEW of BIOEQUIVALENCE

• if two products, containing the same drug, have the same

concentration-time profile at the intestinal membrane surface

then they will have the same rate and extent of absorption

• same in vivo dissolution profile under all luminal conditions

(formulation components do not effect the membrane
permeability and/or intestinal transit)

(Amidon et al.1995)
 Dissolution, solubility, and intestinal permeability are the three
major factors that govern the rate and extent of absorption of a
drug that is stable in the GI tract

Fluid volume
pH Stomach
hydrodynamics
surface tension Small Intestine (major site for absorption)

other….

TIME (hours)
 BCS Class Boundaries: Objectives

Dissolution
(Product)
Very rapid/rapid dissolution - ensure that
in vivo dissolution is not likely to be the
“rate determining” step
Solubility
(Drug) High solubility- ensure that solubility
is not likely to limit dissolution and,
therefore, absorption
Permeability High permeability - ensure that drug
(Drug) is completely absorbed during the limited
transit time through the small intestine
Rapid (and similar) High
Dissolution Solubility

Candidates
for
Biowaivers

High Therapeutic
Permeability Window

low permeable drugs also candidates!

To be noted:

BCS-based ‘Biowaiver’.....
.....is defined as

 in vitro instead of in vivo ‘bioequivalence’ testing

 comparison of test and reference

....is not defined as no equivalence test

Evaluation of drug substance and drug product

Drug substance
 pharmacodynamic / therapeutic aspects
 physicochemical aspects

Drug product
 in vitro dissolution
To be noted:

RISK assessment

♦ “Do not accept critical use medicines”

♦ “Do not accept narrow therapeutic index
drugs”
♦ “take into account documented evidence
for BA or BE problems
♦ “take into account scientific evidence that
API polymorphs, excipients or the
manufacturing process affects BE”
 meaningful literature data may be used
for drug substance characteristics
(and excipients)
 Primary studies.

 product related data must always be actually

generated for the particular product
Class I and III drugs are candidates for a biowaiver:
but what to be addressed?

High solubility
 the highest single unit dose that is completely soluble in 250
ml or less of aqueous solution at pH 1 - 6.8 (37 degrees C)

generate a pH-solubility profile

 possible stability problems have to be considered
 discussion on ‘intermediate solubility’, i.e., pH-
dependent (high) solubility
Solubility
 Generate a pH-solubility profile:

 Equilibrium solubility of the drug substance over the pH

range of 1.2 – 6.8 at 37 ± 1°C.

 At least 3 buffers, including pH 1.2, 4.5 and 6.8

 Solubility at the pKa should be included, if within this range

To be noted:

- pH should be checked at start and end of test:

Example 1:

Example 2:
Example 3:
To be noted:……
 possible stability problems have to be considered

 stability testing:
- dissolve amount of drug in the buffer
- obtain a filtered sample
- store sample at 37±1oC and analyse at certain time
points
- FDA: sample should be at least stable for 1h at pH 1.2 and
at least 3h at pH 4.5 and higher (<5% degradation)
To be noted:

- possible stability problems have to be considered:

Example:
High permeability
♦ at least 85 % absorption in humans
Pharmacokinetic studies in humans:
- Mass balance
- absolute bioavailability
Fraction of Dose Absorbed (F%) Vs.
Permeability (Peff)

Intestinal perfusion methods 100

- Humans (in vivo) 90

80
70
60
High Permeability

- Animals (in vivo or in situ) 50

40
30
20
10
0
0 1 2 3 4 5 6 7 8 9 10

Peff x 10 -4(c m/sec)

In vitro methods using appropriate membranes

- Monolayer of functional cultured human intestinal cells

In case >85% absorption cannot be proven, drug should

be classified as Class III
Permeability:……
 At the moment only in vivo data acceptable as support.
 In vitro (Caco2 cell) only supportive.
 High permeability if:

 Absolute bioavailability is ≥85%

 ≥ 85% of the dose is recovered in urine, as parent and/or Phase I
oxidative and Phase 2 conjugative metabolites
 ≥ 85% of the dose is recovered in urine in feces; feces only
absorbed metabolites.
Formulation aspects: Biowaivers Class I

 Well‑established excipients in usual amounts

 Excipients that might affect the bioavailability of the API,
e.g. mannitol, sorbitol or surfactants, should not differ
qualitatively and must be quantitatively similar between
the test product and comparator product.
 There is some flexibility in the non critical excipients
employed.
 The generic and comparator product are very rapidly
dissolving or similarly rapidly dissolving;
Formulation aspects: Biowaivers Class III (not
included in the ASIAN guideline)

critical parameter permeability;

but to which extent?

less dependent on formulation

often exhibit site-dependent absorption

(transit time may be critical:
dissolution criteria!!)
Formulation aspects: Biowaivers Class III (not
included in the ASEAN guideline)

 ALL excipients in the proposed product formulation should be

qualitatively the same and quantitatively similar to that of the
comparator product
 The generic and comparator product are very rapidly dissolving.

What is similar??
Formulation aspects: similarity
 Not defined EMA/ASEAN guideline.
 WHO in line with SUPAC

 ICH (M09) under discussion

Formulation aspects
Similar dissolution profiles:

 generic vs comparator/reference
 media: pH 1.2, 4.5, and 6.8 (pharmacopeia)
 and pKa
 no surfactants!
 paddle 50 rpm or basket 100 rpm
Formulation aspects
Similarity in profiles:

 both ≥ 85% in 15 minutes (very rapidly)

 both ≥ 85% in 30 min: f2 ≥ 50 (rapidly)
 pre-conditions met e.g. RSD
 note: ICH (M09) proposes only f2.

 Class I: rapidly or very rapidly

 Class III: very rapidly
And the risk for other classes?
need for BE study

LS/LP
Class 4
LS/HP
Class 2
HS/LP
Class 3
HS/HP
Class 1

risk for differences

To be noted:
 Full Dose Combination: biowaiver possible taking into account all
actives.
 Biowaivers in principle only possible for pharmaceutical equivalent
products:
 tablet vs. tablet, capsule vs. capsule
 Biowaiver for a pro-drug only if the pro-drug is absorbed.
 Formulation aspects cannot be taken into account for the metabolite; in
dissolution you measure the parent (pro-drug) and not the metabolite.
 Prevent a biowaiver on a biowaiver.
 If the highest strength is accepted based upon a biowaiver, the additional
strength cannot be accepted based upon a biowaiver for additional
strengths.
 Prevent a biowaiver on a biowaiver.

100 mg biowaiver
100 mg Test
Reference

no biowaiver
no
bio
wa
iv er

50 mg 50 mg; line
extension of
Reference 100 mg Test
biowaiver
 FDA EMA WHO
CLASS I CLASS II CLASS I CLASS II CLASS I CLASS II

Highly Highly Highly Highly Highly Highly

permeable permeable permeable permeable permeable permeable
Highly soluble Low soluble Highly soluble Low soluble Highly soluble Low soluble

Eligible Not Eligible Eligible Not Eligible Eligible Not Eligible

CLASS III CLASS IV CLASS III CLASS IV CLASS III CLASS IV

Low permeable Low permeable Low permeable Low permeable Low permeable Low permeable
Highly soluble Low soluble Highly soluble Low soluble Highly soluble Low soluble

Eligible Not Eligible Eligible Not Eligible Eligible Not Eligible

 ICH (M09) proposes BCS Class I and III

Example: 1.
Example: 2.

Combivir

Lamivudine
Class III
End

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