Chemistry & Biology 14

Supplemental Data
Examination of the Role of Intestinal Fatty
Acid-Binding Protein in Drug Absorption Using
a Parallel Artificial Membrane Permeability Assay
Tony Velkov, James Horne, Aisha Laguerre, Eric Jones, Martin J. Scanlon, and
Christopher J.H. Porter

FENAMATES
O

OH
HO

OH

H
N

HO

Cl
H
N

H
N

CF3

HO

CH3

H
N

CH3

Cl

H
N

CH3

CH3

Cl

Cl

Cl

Tolfenamic Acid
Ki=2.8 0.7 M
logP=5.9; pKa=3.7
logD(7.4)=2.3

Flufenamic Acid
Ki= 3.7 0.8M
logP=5.6; pKa=3.7
logD(7.4)=2.0

Meclofenamic Acid
Ki=8.9 2.0 M
logP=5.9; pKa=4.2
logD(7.4)=2.3

Mefenamic Acid
Ki= 62.810.2 M
logP=5.3; pKa=3.7
logD(7.4)=1.8

Diclofenac
Ki=152.7 9.6 M
logP=3.3; pKa=4.2
logD(7.4)=0.1

PROPIONIC ACID DERIVATIVES

O

CH3

CH3

OH

OH

OH

O
CH3

CH3

OH

Fenoprofen
Ki=14.2 0.5M
logP=3.8; pKa=4.5
logD(7.4)=0.7

Flurbiprofen
Ki= 15.42.8 M
logP=4.1; pKa=4.3
logD(7.4)= 0.9

Ketoprofen
Ki=23.7 3.0 M
logP=2.8; pKa=4.6
logD(7.4)=-0.3
CH3

CH3

CH3

OH

CH3

CH3

Nabumetone
Ki=NB
logP=2.8; pKa=NI
logD(7.4)=2.8

Ibuprofen
Ki= 32.24.2 M
logP=3.7; pKa=4.4
logD(7.4)=0.8

Naproxen
Ki=56.1 4.2 M
logP=3.0; pKa=4.2
logD(7.4)=0.03
O

OH

CH3

CH3

Indoprofen
Ki= 129.11.8 M
logP=1.7; pKa=4.4
logD(7.4)=-1.2

FIBRATES
O
O

CH3

OH
CH3

O
O

Cl

Cl

OH
H
N

OH

CH3

CH3
O

Cl

CH3

Cl

Fenofibric Acid
Ki= 1.00.2 M
logP=3.9; pKa=3.1
logD(7.4)=-0.04

Ciprofibrate
Ki=23.5 1.1 M
logP=2.5; pKa=3.3
logD(7.4)=-1.3

CH3

Bezafibrate
Ki=32.9 0.7 M
logP=3.6; pKa=3.3
logD(7.4)=-0.2

OH

CH3
Cl

CH3

Clofibric Acid
Ki=51.5 1.8 M
logP=2.7; pKa=3.2
logD(7.4)=-1.1

NAPHTHALENE DERIVATIVES

HN

NH

NH

OSO3H

SO3-

O3S

bisANS
Ki=2.00.5 M
logP=5.8; pKa=1.7
logD(7.4)=-2.2

SO3-

H3C

ANS
Ki= 3.20.3 M
logP=?; pKa=1.4
logD(7.4)=-1.3

N
H

TNS
Ki= 45.010.1 M
logP=3.3; pKa=1.0
logD(7.4)=-0.8

BRIDGED BI-PHENYL DERIVATIVES

CH3
O
O
CH3

CH3

O
CH3

CH3

CH3
O

CH3
O

NH

HN

N
CH3

CH3

CH3

Verapamil
Ki=NB
logP=5.0; pKa=8.9
logD(7.4)=3.4

Pyrilamine
Ki=NB
logP=3.3; pKa=9.0
logD(7.4)=1.6

Phenytoin
Ki= NB
logP=2.92; pKa=8.3
logD(7.4)=2.47

CH3
O

CH3

O
O

Cl

N+
OH

CH3

CH3

OH

Cl

Bifenox
Ki=NB
logP=5.8; pKa=NI
logD(7.4)=5.8

Benzyl salicylate
Ki= NB
logP=4.0; pKa=8.1
logD(7.4)=3.9

Mepronil
Ki=NB
logP=4.1; pKa=13.1
logD(7.4)=4.1

NH

H3C

CH3

N-Phenyl-2,3-xylidine
Ki=NB
logP=3.9; pKa=0.8
logD(7.4)=3.9

OTHER MONO-CARBOXYLIC ACID DERIVATIVES

O
OH

OH

Cl

OH

NO2

Acifluorfen
Ki= 4.21.2 M
logP=5.9; pKa=1.9
logD(7.4)=1.9

2-Naphthoxyacetic acid
Ki=7.2 1.9 M
logP=2.6; pKa=3.2
logD(7.4)=-1.3

1-Naphthaleneacetic acid
Ki=32.1 5.7 M
logP=2.7; pKa=4.3
logD(7.4)=-0.3

O
H
O

OH

OH

OH

CH2
C

OH

Benzilic Acid
Ki= 110.813.9 M
logP=3.0; pKa= 3.0
logD(7.4)=-1.1

CH3

Jasmonic Acid
Ki= 140.120.8M
logP=1.7; pKa=4.5
logD(7.4)=-1.2

CH3

OH

CH3

CH3

OH

Asprin
Ki=304.9 10.7 M
logP=1.29; pKa=3.5
logD(7.4)=-2.5

Nalidixic Acid
Ki=NB
logP=0.2; pKa=5.0
logD(7.4)=-2.9

INDOLE ACIDS
N

OH

OH

Indole-3-butyric Acid
Ki=72.3 12.2 M
logP=2.3
pKa=17.4 -NH
4.8 -COOH
logD(7.4)=-0.2

3-Indoleacetic Acid
Ki=92.8 15.6 M
logP=1.4
pKa=16.7 -NH
4.8 -COOH
logD(7.4)=-1.2

HETEROARYL ACETIC ACID DERIVATIVES

O

CH3
O

N
N
O

CH3

HO

Tolmetin
Ki= 134080 M
logP=1.6; pKa=3.5
logD(7.4)=-1.4

OH

Ketorolac
Ki=1670 110 M
logP=0.8; pKa=3.5
logD(7.4)=-1.9

Valproate
Ki=242.723.1 M
logP=2.8; pKa=5.0
logD(7.4)=0.2

CH3

STEROIDS
CH3

CH3

HO
CH3

OH

HO

OH

OH

CH3

OH

OH
CH3

CH3

CH3

CH3

CH3

OH

O
CH3

Progesterone
Ki=20.1 4.6 M
logP=4.0; pKa=NI
logD(7.4)=4.0

Prednisolone
Ki=95.2 9.4 M
logP=1.7; pKa=12.5
logD(7.4)=1.7

Dexamethasone
Ki=1100 109 M
logP=2.1; pKa=12.1
logD(7.4)=2.1

Cortexolone
Ki=1570 89 M
logP=2.7; pKa=12.5
logD(7.4)=2.7

BENZODIAZEPINS

CH3

Cl

H
N

O-

N+

Diazepam
Ki=1980 87 M
logP=2.9; pKa=3.4
logD(7.4)=2.9

H
N

Cl

OH

Cl

Nitrazepam
Ki=2010 95 M
logP=2.8
pKa=10.4
3.02
logD(7.4)=2.8

Lorazepam
Ki=2100 92 M
logP=2.5
pKa=10.8
0.03
logD(7.4)=2.5

TRICYCLIIC DERIVATIVES
CH3
N
CH3
N

N
Cl

N
N

N
N
H

R1

Dibenzazepines
Drug
R1
Imipramine Ki=NB; logP=4.5; pKa=9.5; logD(7.4)=2.4 -(CH2)3N(CH3)2
Desipramine Ki=NB; logP=3.9;pKa=10.2; logD(7.4)=1.2 -(CH2)3NHCH3
Trimipramine Ki=NB; logP=4.8; pKa=9.4; logD(7.4)=2.9 - CH2CH2(CH3)CH2N(CH3)2

Clozapine
Ki=NB
logP=3.5; pKa=8.0
logD(7.4)=2.4

Mianserin
Ki=NB
logP=3.8; pKa=7.1
logD(7.4)=2.8

CH3
N
CH3

R1

Dibenzcycloheptenes
Drug
Amitriptyline Ki=NB; logP=6.1; pKa=9.4; logD(7.4)=4.3
Nortriptyline Ki=NB; logP=5.7; pKa=9.7; logD(7.4)=3.1

R1
=CH(CH2)2N(CH3)2
=CH(CH2)2NHCH3

Dothiepin
Ki=NB
logP=5.6; pKa=10.09
logD(7.4)=3.8

-ADRENERGIC BLOCKERS
OH

CH3

OH
CH3

CH3
O

HN

CH3

CH3

N
H

HN

Metoprolol
Ki=NB
logP=1.8; pKa=9.56
logD(7.4)=-0.4

Atenolol
Ki=NB
logP=0.1; pKa=9.5
logD(7.4)=-2.0

CH3

Acebutolol
Ki=NB
logP=2.6; pKa=9.5
logD(7.4)=0.5

Nadolol
Ki=NB
logP=1.3; pKa=9.7
logD(7.4)=-0.8
CH3

H
N

CH3

N
H

CH3

OH

H
N

CH3

CH3

OH

OH

CH3

H2N

OH

O
CH3

OH

CH3

N
HN

CH3
N

OH

HN

H2N

OH

CH3

H
N

CH3

HN

HO

CH3
H

Timolol
Ki=NB
logP=-0.2; pKa=13.48
logD(7.4)=-2.0

CH3

OH

CH3

Pindolol
Ki=NB
logP=1.9; pKa=9.5
logD(7.4)=-0.2

Labetalol
Ki=NB
logP=2.9
pKa=9.4
7.5
logD(7.4)=0.9

Propranolol
Ki=NB
logP=3.1; pKa=9.5
logD(7.4)=0.9

MICELLANEOUS COMPOUNDS

HO

OH

Cl

OH

CH3

CH3

Nalbuphine
Ki=NB
logP=2.3
pKa=9.6

NH

N
S

N
H

N
H

Cimetadine
Ki=NB
logP=0.4; pKa=6.8
logD(7.4)=0.2

H
N

NH
Cl

Clonidine
Ki=NB
logP=1.5; pKa=8.2
logD(7.4)=-0.8

7.2
logD(7.4)=0.4

NB: No binding detected.

NI: No ionizable groups.

Figure S1. The Chemical Structures of Generic Drugs Examined for I-FABP
Binding Affinity
The I-FABP binding affinity as measured by the ANS fluorescence displacement
assay (under the low salt buffer condition) and physicochemical properties of
each compound are indicated.

Figure S2. The 1H,15N HSQC Spectra of apo-I-FABP (red) Superimposed with
the 1H,15N HSQC Spectra of holo-Drug Bound I-FABP
(A) holo-Fenofibric acid I-FABP, purple (B) holo-Tolfenamic acid I-FABP, yellow
(C) holo-Clofibric acid I-FABP, blue.

Table S1. Membrane Retention Data for the PAMPA Drug Transport Model
Membrane retention at 7h (% initial)
% reduction in
membrane
retention

I-FABP binding
affinity (Ki, M)

4.86 0.7

94.6

0.06 0.01

17.4 3.3

1.03 0.2

94.0

2.8 0.7

Progesterone

83.4 2.7

76.4 5.3

8.39

20 4.6

Ibuprofen

1.8 0.2

0.5 0.1

72.2

58 8.4

Diazepam

42.1 0.6

35.8 3.1

14.9

2000 87

Propranolol

11.5 0.5

11.6 0.9

0.86

NBa

Donor

Buffer

Buffer

Acceptor

Buffer

I-FABP

Oleic Acid

89.9 3.1

Tolfenamic
acid

NB: No binding detected