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1 Neuroscience Therapeutic Area 28 Mar 2014
Influence of positive allosteric modulation of the mGlu2-receptor on the behavioral responses in animal models of depression
Neuroscience Discovery
Janssen Research and Development, a Division of Janssen Pharmaceutica NV,
Luc Ver Donck, J Adriaan Bouwknecht, Hilde Lavreysen
ADAA – Chicago, March 28, 2014
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Disclosure: Luc Ver Donck
• Employment: Janssen Pharmaceutica NV, a Johnson&Johnson company
• Stock shareholder: Johnson&Johnson
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Glutamate
• Primary excitatory neurotransmitter in CNS (Riaza Bermudo-Soriano et al, 2012)
• Activates glutamate receptors (Kew & Kemp, 2005):
– Ionotropic: NMDA, AMPA
– Metabotropic GPCRs (mGlu)
• mGlu receptors (Niswender & Conn, 2010):
– 8 subtypes, 3 main groups
– modulate glutamate tone and phasic release in a subtle manner
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The metabotropic glutamate 2 receptor (mGlu2)
• Pre-synaptic group II receptor, Gi-coupled (Cartmell & Schoepp, 2000)
• Expression in prefrontal cortex, thalamus, striatum, hippocampus, amygdala (Wright et al, 2001, 2013; Lavreysen et al, 2013)
• mGlu2-activation reduces EPSPs: glutamatergic neurotransmission (Kew et al, 2001)
(Ferragutti & Shigemoto, 2006)
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Excessive glutamatergic transmission
• Dysfunctional glutamatergic neurotransmission may be implicated in the pathophysiology of psychiatric illness. (Weinberger, 2007; Olney et al, 1999)
• Hypothesis: therapeutic efficacy by activation of group II mGlu receptors to normalise glutamatergic hyperexcitability. (Fell et al, 2011; Coyle, 2006; Johnson et al, 2005)
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mGlu2 receptor activation
Tateyama et al, 2004 Havlackova et al, 2005
Rondard et al, 2006 Brock et al, 2007
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mGlu2 receptor activation
Tateyama et al, 2004 Havlackova et al, 2005
Rondard et al, 2006 Brock et al, 2007
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mGlu2 receptor activation
Tateyama et al, 2004 Havlackova et al, 2005
Rondard et al, 2006 Brock et al, 2007
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JNJ-40411813
mGlu2 receptor modulation via allosteric site
Tateyama et al, 2004 Havlackova et al, 2005
Rondard et al, 2006 Brock et al, 2007
PAM
• Structurally diverse and subtype-selective, wider chemical space
• Potentially better drug-like properties
• PAMs enhance transmission by endogenous transmitters and preserve temporal pattern of signaling (effective and safer approach?)
Orthosteric agonist
• No subtype-selectivity
• Possibility of over-activation, desensitisation (tolerance)
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JNJ-40411813, an mGlu2 Positive Allosteric Modulator (PAM)
hmGlu2_CHOK1_glutamate_GTPγS; Mean ± SEM, n=17 Lavreysen et al, 2013
• Positive efficacy cooperativity • Minor intrinsic agonism
EC50:147 nM
-9 -8 -7 -6 -5 -4 -3 -2
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
3 .1 0- 6
M J N J -4 0 4 1 1 8 1 3
1 0- 5
M J N J -4 0 4 1 1 8 1 3
c o n tro l
1 0- 8
M J N J -4 0 4 1 1 8 1 3
3 .1 0- 8
M J N J -4 0 4 1 1 8 1 3
1 0- 7
M J N J -4 0 4 1 1 8 1 3
3 .1 0- 7
M J N J -4 0 4 1 1 8 1 3
1 0- 6
M J N J -4 0 4 1 1 8 1 3
g lu ta m a te , L o g (M )
cp
m
control
10-8
3.10-8 10-7 3.10-7 10-6 3.10-6
JNJ (M) 10-5
[Glutamate], Log (M)
cp
m
-1 0 -9 -8 -7 -6 -5 -4
0
5 0
1 0 0
1 5 0
2 0 0
2 5 0
3 0 0 in p re s e n c e o f E C 2 0 G lu
(P A M )
w ith o u t G lu
(A G O )
[J N J -4 0 4 1 1 8 1 3 ] , L o g (M )%
of
ma
xim
al
Glu
re
sp
on
se
no Glu (AG)
+Glu[EC20] (PAM)
[JNJ-40411813], Log (M) %
max G
lu r
esp
on
se
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JNJ-40411813, a PAM at native mGlu2 receptors
Lavreysen et al, 2013
• Acts as PAM at native mGlu2 receptors
No effect in mGlu2 KO-mice
0 1 10
0
1
10
JNJ (µM)
glutamate (µM)
Brain Slices-[35S]GTPγS autoradiography
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JNJ-40411813: target engagement at mGlu2
Lavreysen et al, 2013
• 5HT2A occupancy due to rat-specific metabolite
WT mGlu2 KO
rat
mouse
[3H]JNJ-46281222
% m
Glu
2 o
ccupancy
[JNJ-40411813]plasma (ng/ml)
observed predicted
ex vivo occupancy
EC50:1032 ng/ml
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Rodent tests of despair
• Acute tests:
– Forced swim test
– Tail suspension test
• (sub)-chronic test:
– Dominant/submissive test
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Forced swim test in mice
Key measures:
• Time Immobile
• Distance Moved -30 min
Test
0-6 min
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mGlu2-PAM does not reduce immobility time in forced swim test in mice
NMRI mice, n=10/group. 1w-ANOVA: * p<0.05 vs. vehicle Treatment: 30 min pre-test
FSTM_100430_40411813
0
100
200
300
0 20 0 5 10 20
imipramine 40411813-AAA
Dose (mg/kg s.c.)
Distance
*
(cm)
0
20
40
60
0 20 0 5 10 20
imipramine 40411813-AAA
Dose (mg/kg s.c.)
Immobility
*
(%)
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Tail suspension test in mice
Key measures:
• Time Immobile
• Distance Moved -30 min
Test
0-6 min
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mGlu2-PAM does not reduce immobility time in tail suspension test in mice
NMRI mice, n=15/group. 1w-ANOVA: * p<0.05 vs. vehicle Treatment: 30 min pre-test
TST_100504_40411813-AAA
0
500
1000
0 20 0 5 10 20
Imipramine 40411813-AAA
Dose (mg/kg s.c.)
Distance
* *
0
100
200
0 20 0 5 10 20
Imipramine 40411813-AAA
Dose (mg/kg s.c.)
Immobility
*
* (cm) (sec)
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D/S : dominant/submissive test in rat
• Principle: competition for food between dominant-submissive pair of food deprived rats
Malatynska et al, J Neurosci Meth 165 :175–182,2007
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D/S : dominant/submissive test in rat
• Model of human depression
– Hypothesis: submissive behavior in social animals is related to human depression (Price, 1967)
• Like depressed humans, subordinate rats show: (Malatynska & Knapp, 2005)
– Increased defensive behavior, weight loss
– Major alterations in sleep, eating and active behaviors
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D/S : dominant/submissive test in rat
• Reduction of Submissive Behavior Model:
– Rodent model for depression
• ‘depressed state’ is represented by the behavior of the submissive animal
– Clinically effective antidepressants attenuate submissive behaviour:
• (des)imipramine, fluoxetine
– No effect: neuroleptics, anxiolytics
Malatynska et al, 2002, 2007; Malatynska & Knapp, 2005
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D/S : dominant/submissive test in rat
• Principle: competition for food between dominant-submissive pair of food deprived rats
Malatynska et al, J Neurosci Meth 165 :175–182,2007
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Dominant/submissive: study design
D/S: Dominant/Submissive Male Sprague Dawley rats, age 8-10 w (*) : imipramine after test
Week:
Selection of D/S pairs
(Δ>25%) (n=220)
1 2
Baseline drinking times (n=80)
• 23h food deprivation 5-min D/S test 1h food
3 4 5
Submissive: vehicle/drug Dominant: vehicle (n=10 pairs/group)
+ Daily D/S-test
QD
Key measures:
• Feeding score per rat
• Dominance level
• Treatments: 1h prior to test (*)
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Imipramine reverses submissive behaviour in rat
Mean ± SEM, n= 9 pairs; vehicle = 20% β-OH-cyclodextrin, imipramine: 10 mg/kg IP, QD week 3 through 5 **, *** P<0.01, 0.001 vs. dominance feeding score (student-t test)
IVP-016-10
vehicle
***
** ** **
2 3 4 5
Time (week)
0
50
100
150
200
250
Feeding
score (s)
Dominant
Submissive vehicle
*** ***
0
50
100
150
200
250
Feeding
score (s)
2 3 4 5
Time (week)
imipramine
Dominant
Submissive
imipramine
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JNJ-40411813 reverses submissive behaviour
Mean ± SEM, n= 9 pairs; JNJ-40411813: SC, QD week 3 through 5 *, **,*** P<0.05, 0.01, 0.001 vs. dominance feeding score (student-t test)
2 3 4 5
Time (week)
0
50
100
150
200
250
Feeding
score (s)
***
**
Dominant
Submissive
10 mg/kg 0
50
100
150
200
250
Feeding
score (s)
2 3 4 5
Time (week)
*** ***
*
Dominant
Submissive
20 mg/kg
IVP-016-10
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Imipramine and JNJ-40411813 reduce dominance levels in rat
Mean ± SEM, n= 9 pairs; vehicle (β-OH-cyclodextrin) or drugs, QD week 3 through 5 *,**,*** P<0.05, 0.01, 0.001 vs .dominance level of second week (2w-ANOVA followed by Bonferroni post tests)
IVP-016-10
20% CD
JNJ-40411813 10 mg/kg
JNJ-40411813 20 mg/kg
imipramine 10 mg/kg
-50
0
50
100
150
** *** Dominance
Level (s) -50
0
50
100
150
*
-50
0
50
100
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*
** -50
0
50
100
150
(week) 2 3 4 5 2 3 4 5
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Summary
• No effect of mGlu2-PAM in “acute” stress-related paradigms
• mGlu2-PAM reduces dominance level
– Suggestive for antidepressant effect
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Interpretation of drug effect in D/S test: potential caveats
• No mGlu2-PAM effect observed in (acute) depression paradigms forced swim and tail suspension tests
– Different “state of mind” in sub-chronic D/S-test?
• Increased aggression in submissive animals?
– mGlu2-PAM reduces aggressive behaviour in mice.
• Anxiolytic effects?
– mGlu2-PAM had no effect in elevated plus maze, stress-induced hyperthermia, hole board.
• Change in food intake or appetite
– mGlu2-PAM reduces food intake.
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Conclusions
• The dominant-submissive model differentiates itself from the classical “acute” models of depression.
– Animals are continuously exposed to psychosocial stressor (aggressor stress by the dominant partner)
• Subchronic stress conditions to submissive rat may affect:
– Neurochemical signaling
– Associated behaviour
• Continued treatment with an mGlu2-PAM under this subchronic stress condition:
– Elicited antidepressant-like effects
– Reflected by normalization of the behavioral responses
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Acknowledgements
• Janssen Neuroscience Discovery and Early Development Teams
• Addex Therapeutics
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