human TLE
B
pilocarpine model of epilepsy
sham
control
refractory TLE
§
0
S-Lic (µM)
*
*
*
300 13 (8)
*
* *
100 9 (5)
100
300 15 (12)
300 4 (4)
0
100 6 (2)
*
ns
100 7 (6)
100
*
*
200
30 8 (6)
*
Increase of fast (%)
*
#
30 6 (2)
Increase of fast (%)
ns
200
epileptic
30 14 (5)
A
S-Lic (µM)
Supplementary Figure 1: S-Lic effects on recovery from inactivation. A, S-Lic effects on
recovery from inactivation are dose-dependent in patients with clinically refractory TLE. B, In
animal model of epilepsy, S-Lic shows dose-dependent effects in epileptic animals (red
bars). Asterisks indicate p<0.05, paired t-test, while # and § indicate p<0.05, Man-Whitney U
test. n-numbers for the recorded cells are shown as insets in bars with numbers in brackets
denoting number of patients/animals.
60
40
*
*
20
0
sham control
epileptic
60
ns
40
* * * * * *
20
0
S-Lic 30 6 (4)
S-Lic 100 6 (4)
S-Lic 300 6 (4)
*
ns
B pilocarpine model of epilepsy
S-Lic 30 6 (4)
S-Lic 100 6 (4)
S-Lic 300 6 (4)
ns
S-Lic 30 5 (5)
S-Lic 100 5 (5)
S-Lic 300 5 (5)
Red.of max. firing frequency (%)
human TLE
Red.of max.firing frequency (%)
A
Supplementary Figure 2: Effects of different concentrations of S-Lic on maximal firing
frequency in A, human TLE and B, experimental epilepsy. N-numbers for all experiments
given within the bars with number of patients/ animals in brackets. Asterisks indicate p<0.05.
A
B
Eslicarbazepine
Oxcarbazepine
300
200
100
400
Plasma (nmol/ml)
Plasma (nmol/ml)
400
300
200
100
0
0
0
4
8
12
16
20
24
0
4
8
400
Eslicarbazepine
Oxcarbazepine
300
200
100
0
0
4
8
12
16
20
400
16
20
24
Eslicarbazepine
Oxcarbazepine
300
200
100
0
24
0
4
8
Time (h)
12
16
20
24
Time (h)
F
100
Eslicarbazepine
Oxcarbazepine
75
50
25
0
0
4
8
12
16
20
24
Brain whole volume (nmol/g)
E
Brain whole volume (nmol/g)
12
D
Brain organic phase (nmol/ml)
C
Brain organic phase (nmol/ml)
Eslicarbazepine
Oxcarbazepine
100
Eslicarbazepine
Oxcarbazepine
75
50
25
0
0
4
Time (h)
8
12
16
20
24
Time (h)
Supplementary Figure 3: Plasma (A and B) and brain (C, D, E, F) concentration–time
profiles of eslicarbazepine and oxcarbazepine following a single oral administration of ESL
150 mg/kg (A, C, E) or 300 mg/kg (B, D, F). The levels in brain organic phase reflect the
assumption that eslicarbazepine and oxcarbazepine distribute mainly in the non-aqueous
brain (that represents 20% of whole brain volume). Symbols represent the mean values ±
SEM of four determinations per time point (n = 4 mice).
B
ESL treated 16 (5)
vehicle treated 18 (4)
0
*
* *
40
sham control
vehicle treated
ESL treated
20
0
ESL treated
100
*
sham control
*
60
vehicle treated
*
sham control 12 (3)
Increase of fast (%)
epileptic
200
C
80
reduction of gmax (%)
A
-6
*
sham control
vehicle treated
ESL treated
*
*
-4
-2
0
V1/2,inact (mV)
Supplementary Figure 4: Effects of S-Lic on INaT in control and chronically epileptic
mice. A, Mice were treated with pilocarpine and developed chronic epilepsy. One group
(ESL treated) was treated orally with 300 mg/kg ESL , while another group received vehicle.
Sham control mice received neither pilocarpine nor ESL or vehicle. After in-vivo monitoring,
mice were used for in-vitro patch-clamp recordings. Bargraphs show effects of 300 µM S-Lic
on time course of recovery in hippocampal neurons. B, 300 µM S-Lic reduced maximal
conductance similarly in all groups. C, Changes in the voltage of half maximal inactivation
caused by application of 300 µM S-Lic. Asterisks indicate p<0.05, paired t-test. N-numbers
for all panels given in bars in A.
ESL (150 mg/kg)
S-Lic
OXC
Plasma
Cmax (nmol/ml)
AUC0-t (nmol.h/ml)
Brain (organic phase)
Cmax (nmol/ml)
AUC0-t (nmol.h/ml)
Brain (whole volume)
Cmax (nmol/g)
AUC0-t (nmol.h/g)
ESL (300 mg/kg)
S-Lic
OXC
21425
64851
213
13911
31847
1639164
192
23919
29560
1450116
11116
754115
41158
3554586
12520
1378236
6413
32325
244
16325
8912
766126
274
29751
Supplementary Table 1: Pharmacokinetic parameters for plasma and brain
eslicarbazepine (S-Lic) and oxcarbazepine (OXC) after oral administration of 150 or
300 mg/kg eslicarbazepine acetate (ESL) in the mouse. The levels in brain organic
phase reflect the assumption that eslicarbazepine and oxcarbazepine distribute
mainly in the non-aqueous brain (that represents 20% of whole brain volume).
Dose
(mg/Kg)
Vehicle
10
25
50
100
200
250
400
500
Eslicarbazepine
acetate (ESL)
168.2 ± 11.8
n.d.
160.1 ± 12.6 ns
180.0 ± 0.0
ns
165.9 ± 14.1 ns
142.1 ± 22.8 ns
119.3 ± 22.9 ns
53.1 ± 21.9
***
16.3 ± 8.0
***
Supplementary Table 2. Effects of eslicarbazepine acetate (ESL) on motor performance of
mice expressed as time spent (cut-off: 180 sec) in the Modified Rotarod Test (mean±SEM,
10 mice/group). Compound was given p.o. 60 minutes before the test on day 2. Mean
reference substance (diazepam 8 mg/kg) drop-off time: 54 sec.
Student´s t test: ns = not significant; *** = P < 0.001.
n.d. = not determined.