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In Vitro In Vivo correlation

Prepared by
Hussein Abu-Ghazaleh
R&D Trainee

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 Definition
•  In Vitro In Vivo Correlation (IVIVC) : A mathematical model that predicts
pharmacokinetic parameters(Cmax, AUC) from in vitro dissolution data.
• Once an IVIVC is established for a set of formulations, then in vitro dissolution tests can
be used in place of further bioequivalence studies in the production or modification of
different formulations

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 Applications of IVIVC

• To Waive bioequivalence studies which may be needed for changing :

• Manufacturing site, supplier of raw materials, method of manufacturing, equipment
changes and formulation changes.
• Approval of dosage strength higher or lower than the doses that have been shown to
be safe and effective in clinical trials.
• Approval of another sponsor’s extended-released product even with the same
release mechanism.
• Approval of a formulation change involving a non-release controlling excipient in the
drug product that may significantly affect drug absorption.

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 Levels of IVIVC
• Level A
• Level B
• Level C
• Multiple Level C correlation.

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 Various parameters used in IVIVC depending on the
level.

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 Level A
• Level A is the most recommended correlation by the FDA and is considered the
most informative.
• Level A correlation is generally linear and represents a point-to-point
relationship between in vitro dissolution rate and in vivo input rate, although
nonlinear correlations may be accepted by FDA if found appropriate.
• It is commonly estimated by a two-stage procedure that includes: deconvolution
followed by establishing a link model that correlates the fraction of drug
absorbed to the fraction of drug dissolved.
• Level A should be used when demonstrating an IVIVC relationship for two or
more formulations with different release rates.
• The FDA guidance suggests that two or more formulations with different release
rates (e.g., slow, medium, and fast release formulations differing by 10%)
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 Point to Point Relationship
point-to-point models relate the in vitro
and in vivo amounts at the same time
according to a straight line, i.e. it's relating
the amount released at a specific time to
the amount absorbed at a specific time 

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 Level B correlation
• Level B correlation uses the same data used in Level A, but is based on the
principle of statistical moment analysis.
• The mean in vitro profile of the drug is compared to the mean in
vivo profile
• Level B is the least useful for regulatory purposes because it does not reflect
the actual in vivo plasma level curve, also, different profiles can give the
same parameter values.
• level b correlation isn’t a point-to-point correlation.

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 Level B correlation

• Different profiles could have the

same mean in vivo dissolution time
so this method isn’t useful.

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 Level C
• It establishes a single-point relationship between in vitro dissolution data, and in
vivo pharmacokinetic (PK) parameters (e.g. Cmax, AUC).
• e.g.,% dissolved in 4 hours and Cmax.
• It is not sufficient for obtaining a biowaiver, but Level C can predict Cmax and AUC,
which can help to establish BA and BE
• It is useful in formulation selection and development but not for regulatory purposes
because it does not reflect the full profile

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 Multiple Level C

• Multiple Level C correlation relates one or several PK parameters of interest to the

amount of drug dissolved at several time points.
• It can be as beneficial as Level A correlation because it’s accepted by the authority.
• If multiple Level C correlation is possible, the existence of Level A correlation is also
highly likely and often preferable

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 Multiple Level C

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 IVIVC mathematical methodologies
There are different mathematical methods to establish an IVIVC and they can be classified
into:
The two-stage method which is deconvolution
The One stage method which convolution

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 Deconvolution method
• In the first stage, a deconvolution method is used to estimate the in vivo
absorption or dissolution time course, i.e. fraction absorbed versus time.
• In the second stage, a link model is established between in vivo absorption–
time profile and in vitro dissolution or release profile.
• Then, plasma concentrations are predicted from in vitro release data using
the link model.

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 Steps involved in IVIVC model development
• 1- determine the BCS class of the drug, IVIVC is likely to succeed when
dissolution is the rate-limiting step in absorption.

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 IVIVC correlation expectation for immediate release product based on biopharmaceutic class.

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 Steps involved in IVIVC model development
• 2-data requirements:
• I. vitro data from all formulations
that are investigated.
• Ideally, The most common and the
best way to develop IVIVC
correlation is to make fast,
intermediate, and slow-release
formulations with at least a 10%
difference in dissolution profiles
between the formulations.
• Also, a dissolution method should
be able to discriminate between the
release characteristics of each
formula.

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 Steps involved in IVIVC model development
• 2-data requirements:
•
• II. The developed formulations
should be tested in a PK study to
determine corresponding in
vivo data from a crossover study.
• It’s expected that the slowest
formula has the lowest
bioavailability, and the fastest
formula has the higher
bioavailability, if that doesn’t occur
correlation cant be established

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 Steps involved in IVIVC model development

3- Deconvolution:
The method of deriving in vivo absorption profile for each corresponding formulation from
the plasma concentration profile for comparison with in vitro dissolution data.

Deconvolution

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 Steps involved in IVIVC model development
• 4-Establishes Fraction absorbed vs. fraction dissolved plot

The way to develop the

correlation is trying to take
the dissolution profile and
try to correlate it with the
fraction of drug absorbed
that was got from the plasma
profile by deconvolution and
the relationship between the
individual’s per cent of drug
absorbed and per cent of
drug dissolved will be
established.
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 Steps involved in IVIVC model development

• Determination fraction of
drug absorbed versus the
fraction of drug dissolved
allow investigators to
predict plasma profile from
in vitro dissolution, so
decreasing the need for
more studies during
development and
submission.

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 How to determine if your model is able to predict in vivo
performance from in vitro data?
• When you graph each formulation alone, or you but all formulations
together on one graph you will get the same correlation between the
fraction dissolved and the fraction absorbed.
• If each formula gives you a different mathematical model this means the
method isn’t robust and can’t predict in vivo data accurately
• So the correlation should be the same for all formulations with different
release characteristic

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 Summary

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 Conventional IVIVC generally uses one of three deconvolution methods:

• Model dependent:
• Wagner-Nelson method
• Loo-Riegelman method
• Model independent:
• numerical deconvolution

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 Wagner-Nelson
• Wagner and Nelson developed an equation for calculating the absorption rate
constant (Ka) and the fraction of dose absorbed from plasma drug
concentration-time profile (in vivo data) for the open-compartment model.
• This method is restricted to being applied to drugs that undergo linear
elimination, so it cant be used for non-linear elimination
• It treats the body as a single compartment(ie. one-compartment model drugs)
Thus, this method is not appropriate for drugs that follow multiple-
compartment characteristics.
• It does not assume that the absorption follows zero- or first-order kinetics.
Finally, it has the advantage of being able to calculate the fraction of drug
absorbed over time without requiring IV plasma drug concentration-time data.

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 Loo-Riegelman
• This method is restricted to being applied to drugs that undergo linear
elimination, so it cant be used for non-linear elimination
•  Loo-Riegelman Method takes a compartmental modelling approach, so it is
used for two-compartment model drugs
• it requires concentration-time data from both extravascular and
intravenous administration of the drug to the same subject.

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  Numerical deconvolution
 • These methods are model-independent. It makes no assumptions on the
number of compartments or kinetics of absorption.
• It requires both extravascular and reference data from an oral
solution/immediate release formulation or IV data.
• In addition, these methods assume that the drug undergoes linear
distribution and elimination and is time-invariant.
• Numerical methods also assume that the input site is the same for all
formulations and that the input rate is constant (similar to infusion)
between two-time points.
• Numerical deconvolution may have an advantage in that in vivo dissolution
may be determined, using a program such as PCDCON

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 How to evaluate if we had good correlation or
not?
By comparing observed values and predicted values and see if they are similar
or close enough to each other

Plasma profile Observed plasma profile Predicted

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 Predictability of correlation

• Comparison between predicted bioavailability and observed bioavailability is done and %

P.E is calculated.

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 Evaluation procedures
• Internal predictability: Based on data used to define the IVIVC model.
• External predictability: Based on additional test data sets.
• In the first case you use data from formulations used to develop the correlation.
• And in the second case you use data sets never used to develop the correlation, like from
different study, or different formulation.
• It is recommended to use data from 3 or more formulations with different release rates

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 Internal predictability criteria
• %PE(abs) Cmax and AUC
• According to FDA guidelines, the average absolute %P.E should be
below 10% and %P.E for individual formulation should be below 15%
for the establishment of IVIVC.
• If criteria aren’t met, proceed to an evaluation of external
predictability.

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 External predictability criteria
• %PE (abs) Cmax and AUC
• 10% or less is acceptable
• 10-20% is inconclusive and you need to further evaluations
• Greater than 20% means that the correlation is unacceptable

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 Prediction of plasma concentration

1- convert cumulative dissolution profile

to dissolution rate.
2- use dissolution rate profile with
concentration profile results from unite
input response to predict plasma
concentration-time profile by
convolution technique.
3- compare data from observed plasma
level with predicted plasma level to
assess the predictability of the IVIVC
model.

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 Evaluation of predictability
internal

In this example,
we have three
formulations, all
of which have
individual PE%
below 15%, and
average below
10%.

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 Evaluation of predictability
External
They took the data
from the fasted arm of
the food effect study
using the dissolution
data they had and they
want to see if they
could predict the
plasma profile

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 General Consideration about in vivo studies
1- No restrictions on study design, it can be cross over which is preferred, or
parallel.
2- it is recommended to include a reference treatment such as:
Oral solution, IV solutions, or immediate release product.
3- studies usually are conducted in the fasted state, but when a drug isn’t
tolerated in the fasted state, studies may be conducted in the fed state.

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 General consideration about in vitro studies
• 1- any in vitro dissolution method can be utilized
• 2- the dissolution conditions once defined should be the same for all
formulations tested in the bio study, this means that you can’t use for one
formulation specific conditions and for another formulation use different
conditions
• 3- the preferred dissolution apparatus is USP I OR II, and it should be used at
a compendial recognized speed.
• 4- An aqueous medium either water or buffered solutions not exceeding pH
6.8.
• 5- there is no need to use complex dissolution methods like bio relevant
media to establish the correlation

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 General considerations about in vitro studies
• 6-For poorly soluble drugs, addition of a surfactant may be appropriate
• 7- In general, non-aqueous and hydroalcoholic systems are discouraged.
• 7- dissolution profiles should be obtained from at least 12 units, if less, the robustness of
the method will decrease.
• 8- The coefficient of variation (%CV) for the mean dissolution of a single batch should be
less than 10%.
• 9- dissolution method should be discriminative between formulations with different
release characteristics

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 General considerations
• To establish the IVIVC model you need at least two formulations with
different release rates, however, there is an exception where you can only
use one formulation, if dissolution is independent of dissolution conditions,
for example, if the release characteristics don’t change if the speed of the
apparatus changed or what media used it will remain the same, example for
this formulations is osmotic-controlled release oral delivery system.

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 Software that may be used to establish IVIVC
• Kinetica
• Phoenix WinNonlin Software
https://www.certarauniversity.com/store/668608-102-od-ivivc-toolkit-for-ph
oenix-winnonlin#description
• SASIETS® Software for Analysis of Pharmacokinetic Data
• NONMEM
• STELLA II for convoluting data, making model modifications, or testing
different dosing regimens
• ADAPT II program for simulation and nonlinear regression

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 Back up slides

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 Therapeutic Index of drug
• Narrow therapeutic index drugs
• External predictability is necessary because of the nature of index of the
drug
• Non-narrow therapeutic drugs
• Internal predictability
• External predictability is recommended but not necessary if internal
pred. criteria are met
• This mean that internal predictability is adequate, but if the criteria aren’t
met, then external predictability should be performed.

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 Wagner–Nelson deconvolution.
• applied only to one-compartment drugs
• This method is based on the mass balance theory, where no kinetic model
for the absorption process is assumed.
• WN method does not require IV drug administration, because it assumes
identical elimination rate coefficient (kel) between intra- and extra-vasal
administrations and, therefore kel can be estimated from the final stage of
the oral curve

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 Wagner–Nelson deconvolution.
• Fabs is the fraction absorbed
• At is the drug amount absorbed at time t
• A1 is the drug amount absorbed at infinite time
• Ct is the drug concentration at time t
• Kel is the elimination rate coefficient
• AUCt 0 is the area under the curve from time 0 to time t
• AUC1 0 is the area under the curve from WNtime 0 that
equation to infinity
represents the fraction
absorbed of the bioavailable dose at time t

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 Convolution method
• convolution procedure models the relationship between in vitro dissolution
and plasma concentration in a single step. Plasma concentrations predicted
from the model and those observed are compared directly.
• For these methods, a reference treatment is desirable, but the lack of one
does not preclude the ability to develop an IVIVC.
• Whatever the method used to establish a Level A IVIVC, the model should
predict the entire in vivo time course from the in vitro data

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 Convolution method
one-stage modeling approaches, which directly relate the in vivo release to
the in vitro release.
a drug input profile based on in vitro dissolution data can be solved together
with distribution and elimination profile.
For these methods, a reference administration could be useful, but it is not
mandatory.
The advantage of this method against two stage methods is that the
relationship between the in vitro release and plasma concentrations of the
drug are set in one step, so that the modeling is focused on the ability to
predict the in vivo behavior.

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 Advantages of convolution relative to deconvolution
• The relationship between measured quantities (in vitro release and plasma
drug concentrations) is modeled directly in a single stage rather than via an
indirect two stage approach.
• • The model directly predicts the plasma concentration time course. As a
result: • The modeling focuses on the ability to predict measured quantities
(not indirectly calculated quantities such as the cumulative amount
absorbed). • The results are more readily interpreted in terms of the effect
of in vitro release on conventional bioequivalence metrics. • It is easier to
construct methods that do not require the administration of an IV, oral
solution, or IR reference dose.

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 Convolution vs deconvolution
Convolution
One- stage method
deconvolution
Two-stage method
the most widely used and are mathematical methodology
required by the FDA to establish an IVIVC

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 Convolution vs deconvolution

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 1.Historically, the overall FDA acceptance rate of IVIVC submissions has
been less than 50%. Factors that may contribute to low IVIVC success rates
include:
• Not selecting appropriate formulation amounts and types for IVIVC
development and validation
• Not reviewing exploratory plots to help guide model building and selection
• Not investigating the reasons behind inconclusive predictability, quality, and
richness of input data
• Not choosing enough parameters for parameterization

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 • IVIVC studies for BCS class I and III compounds may not be needed because
these compound classes have the potential for biowaivers.15 The in vivo
dissolution rate of a BCS I must be slower than the gastric emptying for an
IVIVC. BCS III and IV compounds have low permeability (potentially rate
limiting), which reduces the probability of achieving an IVIVC. The case
study presented here is for a BCS IV compound. The highest probability of
achieving an IVIVC for an IR product is a BCS II compound with low solubility
and high permeability

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 In vitro dissolution media
• A common dissolution medium is dearated water, simulated gastric fluid (pH
1.2), or intestinal fluid (pH 6.8 or 7.4) without enzyme, and buffers with a pH
range of 4.5 to 7.5 and be maintained at 37°C. For sparingly water-soluble
drugs, use of surfactants in the dissolution medium is recommended. A
simple aqueous dissolution media is also recommended for BCS Class I drug
as this type of drug exhibits lack of influence of dissolution medium
properties. Water and simulated gastric fluid then are the default mediums
for most of the Class I drugs. A typical medium volume is 500 to 1000 ml

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 notes
Ivivc shouldn’t be linear it may be non linear
Reference treatment is a drug that a the tested product is compared to. The
reference treatment can be iv solution, oral solution, or immediate release
product.
Point to point models are models that relate the amount released at a
specific time to the amount absorbed at a specific time (usually the same
time)

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 IVIVC FOR IR AND XR PRODUCTS

IVIVCs have been developed in the past mostly for controlled-release products,
and the US FDA has a guidance on developing an IVIVC for extended-release (ER)
oral dosage forms. The FDA guidance on dissolution testing of immediate-
release (IR) forms provides generalities about IVIVC development, but not with
as much detail as outlined in the ER guidance.
The development of an IVIVC for an IR oral dosage can be challenging to achieve
because the physiological digestive processes (gastric emptying, hydrodynamic,
and pH variations along the GI tract, etc.) have a significant effect on the IR
dosage form dissolution rate, in addition to absorption parameters. IVIVCs for IR
dosage forms may be achieved if the drug’s dissolution process in vivo is the
rate-limiting step in the overall absorption process.

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 IR VS XR
• a higher degree of correlation may be expected with controlled-release
formulations, since release from the dosage form tends to become the rate-
limiting step in absorption, overcoming permeation rate limitations.
• Also, the time frame allowed to characterize the profile of dissolution or
absorption is much longer than with immediate-release dosage forms,
which permits a greater degree of accuracy and precision in characterizing
the dissolution and absorption profile

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 A number of rational considerations should be taken into account before attempting an in
vitro-in vivo correlation for solid oral dosage forms

• if a drug has a fairly narrow therapeutic window, an in vitro-in vivo correlation may
still not be acceptable as a surrogate for bioequivalency testing
• degree of linearity in pharmacokinetics of the drug may limit degree of correlation,
and variation in the rate and extent of absorption or disposition may limit correlation
• the physical absorption of drug should not be the rate-limiting step in the absorption
process (i.e. is not permeation rate-limited), the release from the dosage form is the
rate limiting step in the overall absorption process (i.e. is dissolution rate-limited)
• dose (mg) and solubility (mg/mL) will determine the volume of dissolution media
necessary to evaluate the formulation in vitro, sink conditions may be difficult to
maintain for large volumes and may indicate problems in vitro and in vivo; however,
in these cases the USP Apparatus III and USP Apparatus IV may be investigated (12).

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 Scaling factor
• If there is a lag time between dissolution and absorption this mean that you
don’t have the correlation, this mean that there is a delay between in vitro
dissolution and fraction absorbed, so in this case its acceptable to use a lag
time to correct and establish the correlation.

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 Reference drug
• plasma concentration data resulting from an IV dose or from the
administration of an IR dosage form, preferably an aqueous oral solution.
More correctly, it requires the results of a dose where the in vivo absorption
or release time course is known, at least approximately.
• unit impulse response: the plasma concentration time course resulting from
the instantaneous in vivo release (or absorption) of a unit amount of drug.

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 goal of an IVIVC
• The ultimate goal of an IVIVC is to establish a meaningful relationship
between in vivo behavior of a dosage form and in vitro performance of the
same dosage form, which would allow in vitro data to be used as a surrogate
for in vivo behavior
• An IVIVC has a mathematical description that provides accurate predictive in
vivo results across the range of dissolution profiles investigated. An IVIVC is
a key parameter in aiding formulation optimization with respect to human
PK response.
•  Once an IVIVC is established for a set of formulations, then in
vitro dissolution tests can be used in place of further bioequivalence studies
in the production or modification of different formulations

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 Parameters used for correlating in vitro dissolution
and in vivo data
In vitro parameters In vivo parameters

Time for a specific amount dissolved Area under the curve

Amount dissolved at a specific a time point Fraction absorbed, absorption rate constant

Mean dissolution time Mean residence time

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 • The linear system analysis is compartment independent model based
on system approach and usually accomplished mathematically by using
the convolution and deconvolution method.
• This method requires the availability of a weighting function for the
human body system i.e. the unit input response (UIR). The convolution
and deconvolution operations are based on two basic assumptions
namely time invariance and superposition.

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