1

Applications of Physiologically Based

Pharmacokinetic Modeling
A Course on Physiologically Based Pharmacokinetic (PBPK)
Modeling and its Applications
March 28
th
- April 1
st
, 2011
Center for Human Health Assessment
The Hamner Institutes for Health Sciences
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3
2
1
0
1
2
3
0 50 100 150
LnConc(uM)
Time(min)
Hepatic
Clearance
Copyright 2011 by The Hamner Institutes for Health Sciences. May not be reproduced without permission
Plasma Protein
Binding
Estimated Renal
Clearance
ReverseDosimetry
Exposure TissueDose
Dose to
Critical Biological Units
Inhalation
Exposure
Effect
Organ
Biologically Based
Dose-Response Modeling
Pharmacokinetics
Inhalation
Ingestion
Dermal
Cancer
Other
Toxicity
Pharmacodynamics
Tissue
Macromolecules
Cells
PBPK
BBDR
2
Role of PBPK Modeling
The purpose of a PBPK model is to define the
relationship between an external measure of
(administered) exposure/dose and an internal
measure of (biologically effective) exposure/dose in ( g y ) p /
both the experimental animal and the human
Physiologically Based Pharmacokinetic Model
C
I
C
X Q
P
Lung
Q
C
C
A
Q
C
Liver
Fat
Q
L
Q
F
Q
R
C
VR
C
VF
C
VL
Rapidly Perfused
(brain, kidneys, etc.)
Slowly Perfused
(muscle, bone, etc.)
Q
S
C
VS
3
Physiologically Based Pharmacokinetic Model
Basis of Description
Model structure
anatomy
QC
Plasma
iv dose
metabolism / transport processes
Model parameters
physiological data (organ weights, blood flows)
biochemical data (partition coefficients,
metabolism)
Model equations
system of mass-balance differential equations
one equation for each tissue
connected by equation for blood
QKid
Kidney
QRap
Rapidly Perfused
QSkn
Skin
QSlw

L L M L L L L A L L
P C K P C V P C C Q dt dA / / / / /
max

connected by equation for blood
E.g., metabolizing tissue (liver):
Slowly Perfused
QLiv
Liver
VMax, KM, KMI
oral dose
Chemical risk assessment
Applications of PBPK Modeling
Chemical risk assessment
Drug development research and evaluation
Interpretation of human biomonitoring data
In vitro to in vivo extrapolation In vitro to in vivo extrapolation
Evaluation of early-life susceptibility
4
Assess the biological determinants that governthe
How are PBPK Models Used
in Human Health Risk Assessment?
Assess the biological determinants that govern the
kinetic behavior
Calculate tissue dose metrics for risk assessment
calculations
Support extrapolation across dose-routes, between
species, from high to low dose levels, and over
various dosing scenarios
Assess mechanisms of response (PD) based on
their relationship with dose metrics for target tissues
Use of PBPK Modeling in
Human Health Risk Assessment by
EPA and Other Agencies
Methylene Chloride (EPA OSHA ATSDR Health Canada) Methylene Chloride (EPA, OSHA, ATSDR, Health Canada)
2-Butoxy Ethanol (EPA, Health Canada)
Vinyl Chloride (EPA)
Chloroform (Health Canada)
Perchlorate (EPA)
Acrylamide (EPA)
Trichloroethylene (EPA)
Perchloroethylene (EPA)
5
Example of the Use of a PBPK Model to
Improve Dosimetry in a Risk Assessment:
Vinyl Chloride
Cross-species correspondence of a rare tumor type:
liver angiosarcoma in mouse, rat, and human
Carcinogenic at doses with no evidence of enhanced cell
proliferation, receptor interaction, or cytotoxicity
Mode of Action Information
Mutagenic; metabolized to reactive intermediates
associated with DNA adduct formation and mistranscription
Expect linear dose-response below the experimental range
Vinyl Chloride Vinyl Chloride Vinyl Chloride
Metabolism of Vinyl Chloride
Chloroethylene
Epoxide
P450
EpoxideHydrolase
DNA Adducts CO
2
H
2
O
GSH
Chloroethylene
Epoxide
Chloroethylene
Epoxide
P450
EpoxideHydrolase
DNA Adducts DNA Adducts CO
2
CO
2
H
2
O
GSH
Chloroacetaldehyde Tissue Adducts
Glutathione
Conjugates
GSH
Chloroacetaldehyde Chloroacetaldehyde Tissue Adducts Tissue Adducts
Glutathione
Conjugates
Glutathione
Conjugates
GSH
Dose metric: concentration (AUC) of chloroethylene epoxide
6
QP
QC
QF
Lungs
Fat
CI CX
CVF
CA
QP
QC
QF
Lungs Lungs
Fat Fat
CI CX
CVF
CA
PBPK Model for Vinyl Chloride
QS
QR
QL
Rapidly Perfused Tissues
Slowly Perfused Tissues
Liver
CVR
CVS
CVL
CA
CA
VMAX2
KM2
VMAX1
KM1
CA
KZER
KA
QS
QR
QL
Rapidly Perfused Tissues Rapidly Perfused Tissues
Slowly Perfused Tissues Slowly Perfused Tissues
Liver Liver
CVR
CVS
CVL
CA
CA
VMAX2
KM2
VMAX1
KM1
CA
KZER
KA
Reactive Metabolites CO
2
Glutathione Conjugate
Tissue/DNA Adducts
KCO
2
KGSM
KM2 KM1
KFEE
GSH
KGSM
KS KO
KA
KB
Reactive Metabolites Reactive Metabolites CO
2
CO
2
Glutathione Conjugate Glutathione Conjugate
Tissue/DNA Adducts Tissue/DNA Adducts
KCO
2
KGSM
KM2 KM1
KFEE
GSH GSH
KGSM
KS KO
KA
KB
Dose metric: production of reactive metabolites per gram liver
(Clewell et al. 2001)
Model Parameterization
Physiological Parameters (from literature)
i l bl d fl tissue volumes, blood flows
alveolar ventilation, cardiac output
Partition coefficients (measured in vitro)
Rodent: blood:air and tissue:air
Human: blood:air
Metabolism (estimated by fitting in vivo data)
Rodent: closed chamber gas uptake, disposition studies
H l d h b t k Human: closed chamber gas uptake
7
10000
Estimation of Metabolism Parameters
Gas Uptake Studies in Male F344 Rats
100
1000
C
h
a
m
b
e
r

C
o
n
c
e
n
t
r
a
t
i
o
n

(
p
p
m
)
1
10
0 1 2 3 4 5 6
Hours
C
250 ppm
550 ppm
1250 ppm
3200 ppm
8000
10000
Estimation of Metabolism Parameters
Radiolabel Studies in Male Sprague-Dawley Rats
4000
6000
8000
T
o
t
a
l

A
m
o
u
n
t

M
e
t
a
b
o
l
i
z
e
d

(
m
g
)
0
2000
0.1 1 10 100 1000 10000
Concentration (ppm)
T
8
10
KM1=0.1
KM1=1.0
Estimation of Metabolism Parameters
Human Inhalation Study Subject A
1
C
h
a
m
b
e
r

C
o
n
c
e
n
t
r
a
t
i
o
n

(
p
p
m
)
0.1
0 0.1 0.2 0.3 0.4 0.5
Hours
10
KM1=0.1
KM1=10
Estimation of Metabolism Parameters
Human Inhalation Study Subject B
1
C
h
a
m
b
e
r

C
o
n
c
e
n
t
r
a
t
i
o
n

(
p
p
m
)
KM1=1.0
0.1
0 0.1 0.2 0.3 0.4 0.5
Hours
C
9
Human risk estimates (per million) for lifetime exposure
to 1 ppb vinyl chloride in air based on the incidence
Cross-Species and Cross-Route
Correspondence Using PBPK Dose Metric
of liver angiosarcoma in animal bioassays
Animal Bioassay Study 95% UCL Risk / million / ppb
Males Females
Maltoni - Mouse Inhalation 1. 52 3. 27
Maltoni - Rat Inhalation 5. 17 2. 24
Feron - Rat Diet 3. 05 1. 10
Maltoni - Rat Gavage 8. 68 15. 70
Comparison of Cancer Risk
Estimates for Vinyl Chloride
Basis Inhalation Drinking Water
Old EPA -- Animal
(mg/kg/day -- BSA)
PBPK -- Animal
(1 ug/m
3
)
84.0 x 10
-6
2.7 x 10
-6
(1 ug/L)
54.0 x 10
-6
1.1 x 10
-6
PBPK -- Human
(Epidemiology)
1.7 x 10
-6
10
Applications of PBPK Models in Drug
Development Research and Evaluation
Integrate information from different studies
Different routes of exposure
Different species
Different dosing regimens
Provide a validated platform for predictive simulation of
alternative dosing methods drug-drug interactions (DDI)
Improve understanding of PD by relating effect to dose at
target tissue or binding to target protein (PBPK/PD)
Support more accurate estimation of equivalent human dosing
to achieve same dose to target protein as in test animals g p
Predict fetal exposure and lactation transfer
Estimate variability of PK across special populations
Obese, Elderly, Infants, Diseases
Polymorphisms
Use of PBPK Modeling to Optimize
the Dosage Regimen of
an Antiparasitic Prodrug
Zhixia (Grace) Yan
Division of Pharmacotherapy and Experimental Therapeutics
UNC Eshelman School of Pharmacy
11
Prodrug X demonstrated reversible hepatotoxicity
in humans
Prodrug X is rapidly metabolized to the active
metabolite Y in the liver metabolite, Y, in the liver
The active metabolite, Y accumulated significantly
in the liver
C ld th t i it h b id d Could the toxicity have been avoided
using a more rational dosage regimen?
Dosage Optimization Strategy
Characterize the hepatobiliary disposition of
pafuramidine and furamidine pafuramidine and furamidine.
Develop and validate a whole-body
physiologically-based pharmacokinetic (PBPK)
model for pafuramidine and furamidine in rats.
Develop a whole-body PBPK model for
f idi d f idi i h pafuramidine and furamidine in humans.
Optimize dosage regimen based on the human
PBPK model.
12
1
PBPK Predicted Prodrug X/Active metabolite Y
Disposition in Human Plasma
l
a
s
m
a

0.1
1
X
Y
0 001
0.01
0.1
C
o
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c
e
n
t
r
a
t
i
o
n

i
n

P
(

M
)
0.001
0.01
0 6 12 18 24
0.001
0 48 96 144 192 240
Terminal t
1/2
(h) Observed Predicted
X 11 19
Y 14 64 (2.5 d)
Time (h)
3x
An Optimized Dosage Regimen for
Efficacy and Safety
1000 1000
t
i
o
n
100 mg X twice daily
40 mg Y once daily
Liver
AUC
0 14d
5x
0.1
1
10
100
0.1
1
10
100
d
i
n
e

C
o
n
c
e
n
t
r
a
t
(

M
)
Liver
AUC
0-14d
5x
NOAEL
0.001
0.01
0 2 4 6 8 10 12 14
0.001
0.01
0 2 4 6 8 10 12 14
Time (d)
F
u
r
a
m
i
d
C
eff,min
Plasma
13
Application of PBPK Modeling for the Interpretation
of Human Biomonitoring Data
Chemical concentrations Chemical concentrations
Margin of safety
Chemical concentrations
in human blood from
biomonitoring studies
Chemical concentrations
in animal blood in
toxicity studies
Pharmacokinetic
modeling
Pharmacokinetic
Modeling
F
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w
a
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d

d
o
s
i
R
e
v
e
r
s
e

d
o
s
i
Human exposures
(Chemical concentrations in
environment)
Animal exposures
(Administered doses in
toxicity studies)
Traditional risk assessment
m
e
t
r
y
m
e
t
r
y
Exposure Reconstruction Using a PBPK Model
Iraqi woman exposed during pregnancy
to grain contaminated with methylmercury
Linking Internal Dose to Health Outcome:
450 6
Maternal
E
42 g/kg/day
200
250
300
350
400
M
e
H
g

i
n

H
a
i
r

(
p
p
m
)
3
4
5
M
e
H
g

i
n

B
l
o
o
d

(
p
p
m
)
Maternal hair
Maternal blood
Infant blood
Exposure
108 days
(Clewell et al., 2000)
0
50
100
150
0 200 400 600 800
Days
M
e
0
1
2
M
e
Pregnancy
14
QSAR
In Vitro
Kinetics
Partitioning
P i l
Application of PBPK Modeling for
In Vitro to In Vivo Extrapolation
PBPK
Model
In Vitro
Dynamics
Partitioning
Metabolism
etc.
Potential
Target Tissues
Target Tissue
Responses
In Vivo
Exposure Profile
In Vivo
Human
Toxicity
Estimate
Nature of
Toxicity
In Vivo
Dose-Response
IVIVE Example:Impact of CYP2C9 Polymorphism
on Coumaden (Warfarin) Internal Dose
QC
Plasma
iv dose
QKid
Kidney
QRap
Rapidly Perfused
QSkn
Skin
(Gentry et al., 2002)
QSlw
Slowly Perfused
QLiv
Liver
VMax, KM, KMI
oral dose
15
Intrinsic
Clearance
Allele Reference Mean CV Mean CV (VmaxC/Km)
CYP2C9*1 Haining et al ., 1996
1
1.61 1.85 0.87
Km (mg/L) Vmax (mg/hr/kg
3/4
)
Metabolic Parameters for (S)-Coumaden
for Three CYP2C9 Alleles
Takahashi et al ., 1998b
2
2.13 0.0036 0.81 0.12 2.6
Sullivan-Klose et al ., 1996
2
1.01 0.046 3.57 0.078 0.28
Rettie et al ., 1994
3
3.2 1.26 2.5
Rettie et al ., 1994
4
3.2 1.05 3
CYP2C9*2 Sullivan-Klose et al ., 1996
2
1.26 0.031 3.85 0.056 0.33
Rettie et al ., 1994
3
0.21 0.52 0.4
Rettie et al ., 1994
4
0.36 0.65 0.55
Rettie et al ., 1999
5
1.1 0.036 1.85 0.17 0.59
CYP2C9*3 Haining et al ., 1996
1
0.31 9.24 0.034
Takahashi et al ., 1998b
2
0.51 0.22 3.2 0.16 0.16 ,
Sullivan-Klose et al ., 1996
2
1.37 0.044 28.4 0.059 0.048
1
baculovirus/insect cell system, purified enzyme
2
yeast expression, microsomes
3
Hep G2 cells, cell lysate
4
Hep G2 cells, particulate preparation
5
expressed in insect cells, purified enzymes
(Haber et al., 2002)
Average Prevalence of CYP2C9 Alleles
in the U.S. Population
Prevalence
S1 homozygous 78%
S1/S2 heterozygous 12%
S1/S3 heterozygous 9%
S2 homozygous 1%
(Haber et al., 2002)
S2/S3 heterozygous 1%
S3 homozygous 0.5%
16
Simulation of impact of genetic
polymorphism on Coumaden internal dose
A . C Y P 2 C 9 * 1 A l l e l e
1
2
3
4
5
a
s
m
a
C
o
n
c
e
n
tra
tio
n
(m
g
/L
)A . C Y P 2 C 9 * 1 A l l e l e
1
2
3
4
5
a
s
m
a
C
o
n
c
e
n
tra
tio
n
(m
g
/L
)
0
0 1 0 0 0 2 0 0 0 3 0 0 0 4 0 0 0 5 0 0 0 6 0 0 0
H o u r s
P
la
B . C Y P 2 C 9 * 2 A l l e l e
0
1
2
3
4
5
0 1 0 0 0 2 0 0 0 3 0 0 0 4 0 0 0 5 0 0 0 6 0 0 0
H o u r s
P
la
s
m
a
C
o
n
c
e
n
tra
tio
n
(m
g
/L
)
C . C Y P 2 C 9 *3 A lle le
5
g
/L
)
0
0 1 0 0 0 2 0 0 0 3 0 0 0 4 0 0 0 5 0 0 0 6 0 0 0
H o u r s
P
la
B . C Y P 2 C 9 * 2 A l l e l e
0
1
2
3
4
5
0 1 0 0 0 2 0 0 0 3 0 0 0 4 0 0 0 5 0 0 0 6 0 0 0
H o u r s
P
la
s
m
a
C
o
n
c
e
n
tra
tio
n
(m
g
/L
)
C . C Y P 2 C 9 *3 A lle le
5
g
/L
)
(Gentry et al., 2002)
0
1
2
3
4
0 1 0 0 0 2 0 0 0 3 0 0 0 4 0 0 0 5 0 0 0 6 0 0 0
H o u r s
P
la
s
m
a
C
o
n
c
e
n
tr
a
tio
n
(
m
g
0
1
2
3
4
0 1 0 0 0 2 0 0 0 3 0 0 0 4 0 0 0 5 0 0 0 6 0 0 0
H o u r s
P
la
s
m
a
C
o
n
c
e
n
tr
a
tio
n
(
m
g
Simulation of Impact of Genetic Polymorphism
on Warfarin Internal Exposure
200
250
A
U
C
Case 2
Case 3
Normal population
Total population
Simulation of impact of genetic
polymorphism on coumaden internal dose






50
100
150
F
r
e
q
u
e
n
c
y

o
f

(
S
)
-
W
a
r
f
a
r
i
n

A
F
r
e
q
u
e
n
c
y










0
0
5
0
1
0
0
1
5
0
2
0
0
2
5
0
3
0
0
3
5
0
4
0
0
4
5
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5
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5
0
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0
0
8
5
0
9
0
0
9
5
0
1
0
0
0
(S)-Warfarin AUC
(Gentry et al., 2002)
(S)-Coumaden AUC
17
Application of PBPK Modeling to
Investigate Age-Dependent Susceptibility
An age-dependent PBPK model was developed to
simulate the physiological and biochemical changes in
humans associated with growth and aging.
All physiological and biochemical parameters in the
model change with time based on empirical data; only
the chemical specific parameters remain constant.
This model was used to simulate blood concentrations This model was used to simulate blood concentrations
of nicotine and its metabolite cotinine for a constant
daily oral dose of 1 mg/kg/day nicotine from birth to 75
years
Age-Dependent Internal Exposure to Ingested Nicotine (1 ug/kg/day)
4 . 0 E - 4 2 . 5 E - 3
N i c o ti n e
C o ti n i n e
1 . 0 E - 4
2 . 0 E - 4
3 . 0 E - 4
B
l
o
o
d

C
o
n
c
.

o
f

N
i
c
o
t
i
n
e

(
m
g
/
L
)
5 0 E 4
1 . 0 E - 3
1 . 5 E - 3
2 . 0 E - 3
B
l
o
o
d

C
o
n
c
.

o
f

C
o
t
i
n
i
n
e

(
m
g
/
L
)
0 . 0 E +0
0 2 0 4 0 6 0 8 0
A g e ( y e a r s )
0 . 0 E +0
5 . 0 E - 4
18
Clewell, H.J., Andersen, M.E., and Barton, H.A. 2002. A consistent approach for the
application of pharmacokinetic modeling in cancer and noncancer risk assessment.
Environmental Health Perspectives 110:85-93.
Clewell, H.J., Gearhart, J.M., Gentry, P.R., Covington, T.R., VanLandingham, C.B., Crump,
K.S., and Shipp, A.M. 1999. Evaluation of the uncertainty in an oral Reference Dose for
methylmercury due to interindividual variability in pharmacokinetics Risk Anal 19:547-
References
methylmercury due to interindividual variability in pharmacokinetics. Risk Anal 19:547-
558.
Clewell, H.J., Gentry, P.R., Covington, T.R., Sarangapani, R., and Teeguarden, J.G. 2004.
Evaluation of the potential impact of age- and gender-specific pharmacokinetic differences
on tissue dosimetry. Toxicol. Sci. 79:381-393.
Clewell, H.J., Gentry, P.R., Gearhart, J.M., Allen, B.C.,and Andersen, M.E. 2001.
Comparison of cancer risk estimates for vinyl chloride using animal and human data with
a PBPK model. Science of the Total Environment 274(1-3):37-66.
Clewell, H.J., Reddy, M.B., Lave, T., and Andersen, M.E. 2007. Physiologically based
pharmacokinetic modeling In: Gad SC ed Preclinical Development Handbook John pharmacokinetic modeling. In: Gad, SC, ed. Preclinical Development Handbook. John
Wiley and Sons, Hoboken, NJ.
Clewell, H. J., Tan, Y. M., Campbell, J.L., and Andersen, M.E. 2008. Quantitative
interpretation of human biomonitoring data. Tox. Appl. Pharmacol., 231:122-133.
Gentry, P.R., Hack, C.E., Haber, L., Maier, A., and Clewell, III, H.J. 2002. An Approach for
the Quantitative Consideration of Genetic Polymorphism Data in Chemical Risk
Assessment: Examples with Warfarin and Parathion. Toxicol Sci 70:120-139.
Rowland M, Balant L, and Peck C. 2004. Physiologically Based Pharmacokinetics in
Drug Development and Regulatory Science: A Workshop Report, AAPS PharmSci. 6(1):E6.