Assessment of the Physiologically-Based iDEATM Predictive Model Using an External (Blinded) Data Set
G. Leesman1, D. Norris1, G. Timony1, Y. Chen1, W. Pirson2, F. P. Theil2, A. H. Schmitt-Hoffman2, Y. H. Lee1, J. Doerr-Stevens1, P. Gordon1, R. Retajczyk1, S. Tran1, K.-J. Lee1, N. Johnson1, J. Castelo1, R. Christopher1, G. Grass1, P. Sinko1
Caco-2 Permeability:
Directionality: AB and BA
Caco-2: 20-23 day/ Passage 30-40/Surface Area: 0.31 cm2
Donor conc: 100 µM including 1% DMSO
A: 300 µL pH 7.4 (or pH 6.5 Mes)/ B: 1200 µL pH 7.4 Ringers buffer
Donor side sampling: 20 µL at beginning and final
Receiver side sampling: 100 µL at 30, 50, 70, and 90 min
Incubation at 50 oscillations per minute, 37 °C, 5%CO2, 95% humidity
N=4 wells per compound
Analysis: LC-MS, HPLC-UV, or LSC
Peff (cm/sec) = (dX/dt)/(A*Co*60), where X: mass transported, A: surface area and Co: initial donor drug concentration
Monolayer integrity: Pre and post-test TEER measurements
Transport markers: Atenolol, propranolol
Rabbit Permeability Model
FDp Predictions Rabbit
RESULTS
Solubility Profiles:
100
90
80
70
60
The physiologically-based iDEA predictive model was trained using a database of 56 non-metabolized compounds and a total of 85 drug-dose
combinations. See Presentation Number: 3523 (Timony et al.) for details. The in vivo training data was obtained from clinical pharmacokinetic studies
provided by Genentech, Parke-Davis, Schering-Plough, SmithKline Beecham, Trega Biosciences and others. The in vitro data inputs for the model were
permeability from rabbit intestinal or Caco-2 and solubility determined at various pH values.
F. Hoffman-La Roche, Ltd. supplied the external validation compound set in a blinded fashion. The external set of drugs was a set of eight compounds
with diverse properties. These drugs were not used to develop the model.
pH 1.5
pH 5.0
A
ng/mL
75.93  23.85
37.74  7.46
B
mg/mL
>100
>100
C
mg/mL
>100
>100
D
mg/mL
0.008  0.001
0.154  0.050
E
mg/mL
25.91  0.77
24.39  0.63
G
mg/mL
>100
>100
H
mg/mL
0.554
3.576  0.154
I
 g/mL
0.97  0.13
1.59  0.04
pH 6.5
pH 7.0
pH 7.5
22.70  8.76
40.00  39.04
21.76  14.25
>100
>100
>100
>100
>100
>100
0.548  0.274
0.764  0.400
0.799  0.017
23.86  0.94
22.70  2.54
24.03  0.63
>100
>100
>100
0.547  0.010
0.552  0.009
0.651 0.011
20.08  8.65
96.90  4.57
433.7  156.0
50
40
30
20
10
0
Jejunum
30
15
A to B
B to A
10
5
25
20
A to B
15
B to A
10
5
30
25
20
A to B
15
B to A
10
0
0
A
B
C
D
E
G
H
I
5
B
C
D
E
G
H
I
Roche Compounds
Roche Compounds
A
B
C
A  B Permeability
• Compounds B, D and H: >5 x 10 -6 cm/sec (high permeability)
•
•
•
•
•
•
•
•
•
•
•
• Compounds C and G: 1 - 5 x 10 -6 cm/sec (intermediate permeability)
• Compounds A, E, and I: <1 x
10 -6
cm/sec (low permeability)
Transport processes
• Passive transport (AB  BA): Compounds D and H
• Facilitated transport
- Influx transport (AB > BA): Compounds A, B, C
- Efflux transport (AB< BA): Compounds E, G and I
D
E
G
Roche Compounds
Rabbit Intestinal Permeability:
Directionality: AB and BA
Male New Zealand white rabbits (2.5-3 kg) (overnight fasting)
Intestinal Segments: duodenum, jejunum, ileum, and descending colon
Ussing side-by-side diffusion chamber/ 37 °C/ gas lift mechanism using 1-2 mL/min 95% O2 and 5% CO2
Donor conc: 100 µM including 1% DMSO
Apical and Basolateral: 1.5 mL pH 7.4 Ringers buffer
Donor sampling: 100 µL at beginning and final
Receiver sampling: 100 µL at 30, 50, 70, and 90 min
N=3 chambers per compound
Analysis: LC-MS/ HPLC-UV/ LSC
Peff (cm/sec) = (dX/dt)/(A*Co*60), where X: mass transported, A: surface area, 0.636 cm2 and Co: initial donor drug concentration
40
20
15
A to B
B to A
10
5
H
I
35
30
25
A to B
20
B to A
15
A
B
C
D
E
G
Roche Compounds
H
I
2
1.5
1
0.5
1
20
40
60
80
0
1
100
10
100
1000
10000
0
0.5
1
1.5
2
2.5
3
3.5
Cmax Predictions Caco-2
100
90
80
70
60
50
40
30
20
10
0
tmax Predictions Caco-2
3
10000
2.5
1000
100
10
20
40
TM
iDEA
60
Predicted FDp
80
100
1.5
1
0.5
1
0
2
0
1
10
100
TM
iDEA
1000
Predicted Cmax (ng/mL)
10000
0.5
1
1.5
2
2.5
iDEATM Predicted tmax (hr)
5
A
B
C
D
E
G
Roche Compounds
H
I
4
iDEATM Predicted tmax (hr)
iDEATM Predicted Cmax (ng/mL)
10
0
0
0
A
Permeability (E-6 cm/sec)
20
Caco-2 at pH 7.4/pH 7.4
25
35
Permeability (E-6 cm/sec)
Simulated gastric fluid (pH 1.5, NaCl/HCl)
Simulated intestinal fluid (pH 5.0, 6.5, 7.0 and 7.5; KH2PO4/NaOH)
N = 3 determinations for each pH
Incubate at 37 oC (rotation), 4 hours
pH adjust, if necessary, and incubate additional 2 hours
Filter aliquot (0.45 M)
Analysis: HPLC/UV
Permeability (E-6 cm/sec)
•
•
•
•
•
•
•
Permeability (E-6 cm/sec)
25
Colon
Ileum
Permeability (E-6 cm/sec)
Duodenum
Solubility Testing:
Caco-2 Permeability
10
FDp Predictions Caco-2
Known FDp
Rabbit Intestinal Permeability
METHODS
100
3
2.5
Caco-2 Permeability Model
• Low Solubility (<1 mg/mL): Compounds A, D, H (except pH 5) and I
Permeability Profiles:
1000
iDEATM Predicted FDp
• High Solubility (>90 mg/mL): Compounds B, C and G
The model was evaluated on its ability to predict both rate and extent of absorption. Extent of absorption was compared by FDp (Fraction Dose absorbed
into the portal vein at 24 hours. The rate of absorption was evaluated by comparing predicted and observed Cmax and tmax. Predicted Cmax and tmax were
determined by linking the output of the absorption model to the appropriate pharmacokinetic distribution model.
4
3.5
Known C max (ng/mL)
Compound
tmax Predictions Rabbit
10000
0
INTRODUCTION
Cmax Predictions Rabbit
Known t max (hr)
•
•
•
•
•
•
•
•
•
•
•
•
Performance of the physiologically-based iDEA predictive models:
Known t max (hr)
Purpose. To evaluate, in a blinded manner, the performance of the physiologically-based iDEA predictive models using a set of eight diverse
compounds, which were not used to develop the model. Methods. Physiologically based absorption models were trained using a database of 56 nonmetabolized compounds and a total of 85 drug-dose combinations. The in vivo training data was obtained from clinical pharmacokinetic studies provided
by Genentech, Parke-Davis, Schering-Plough, SmithKline Beecham, Trega Biosciences and others. The in vitro data inputs for the model were rabbit
intestinal permeability from various regions or Caco-2 permeability and solubility determined at various pH values. F. Hoffmann-La Roche, Ltd.
supplied the external validation compound set in a blinded fashion. The compounds were very diverse in terms of structure, solubility (22 ng/mL to >100
mg/mL), rabbit permeability (0.14 to 25 x 10-6 cm/s), Caco-2 permeability (0.43 to 32.5 x 10-6 cm/s) , and pharmacokinetic properties. The acceptance
criteria for accuracy of the predictions were established for fraction of dose absorbed into the portal vein (FDp), Cmax and tmax prior to the evaluation.
Results. The rabbit permeability model predictions for FDp, which ranged from 4 to 100%, were highly correlated (r2 = 0.88, absolute mean error =
8.8%) with the observed values, with all 8 compounds predicted within the acceptance criteria. The rabbit permeability model predictions for Cmax and
tmax could be evaluated for only seven of the compounds. The rabbit permeability model accurately predicted Cmax for 5 of 7 drugs and tmax for 6 of 7
drugs. The Caco-2 permeability model predictions for FDp were also highly correlated (r2 = 0.85, absolute mean error = 12.9%) with the observed
values, 7 of 8 compounds predicted within the acceptance criteria. The Caco-2 permeability model accurately predicted Cmax for 6 of 7 drugs and tmax for
6 of 7 drugs. Conclusions. The physiologically-based IDEA predictive models accurately predicted relevant biopharmaceutical outcomes for a diverse
blinded external validation set of eight drugs.
Biosciences, Inc.: San Diego, CA United States ; 2F. Hoffmann-La Roche Ltd.: Basel, Switzerland
Known FDp
ABSTRACT
1Trega
Known C max (ng/mL)
trega
CONCLUSION
• The physiologically-based iDEA predictive model has been trained using a database of 56 non-metabolized compounds and a total of
85 drug-dose combinations.
• A set of eight diverse drug compounds provided by F. Hoffmann-La Roche, Ltd. Were processed through in vitro solubility and permeability assays.
The results from those assays were used for evaluating the performance of the physiologically-based iDEA predictive models in a blinded fashion.
• This external set of drugs varied in their physical and chemical properties: MW: 200 to 552, solubility (22 ng/mL to >100 mg/mL), permeability
(Rabbit: 0.14 to 25  10-6 cm/s; Caco-2: 0.43 to 32.5  10-6 cm/s), transport process and pharmacokinetic properties (%FDp 4 to 100%).
• iDEA predictive models accurately predicted the relevant biopharmaceutical outcomes (FDp, Cmax and tmax) for a diverse blinded external
validation set of eight drugs.
3