Influence of positive allosteric modulation of the mGlu2 ... Donck_310.pdfInfluence of positive allosteric modulation of the mGlu2-receptor on the behavioral responses in animal models

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


Disclosure: Luc Ver Donck

• Employment: Janssen Pharmaceutica NV, a Johnson&Johnson company

• Stock shareholder: Johnson&Johnson


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


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)


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)


mGlu2 receptor activation

Tateyama et al, 2004 Havlackova et al, 2005

Rondard et al, 2006 Brock et al, 2007










JNJ-40411813

mGlu2 receptor modulation via allosteric site



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)


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


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


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


Rodent tests of despair

• Acute tests:

– Forced swim test

– Tail suspension test

• (sub)-chronic test:

– Dominant/submissive test


Forced swim test in mice

Key measures:

• Time Immobile

• Distance Moved -30 min

Test

0-6 min


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

*

(%)


Tail suspension test in mice

Key measures:

• Time Immobile

• Distance Moved -30 min

Test

0-6 min


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)


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



• 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



• 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



• Principle: competition for food between dominant-submissive pair of food deprived rats

Malatynska et al, J Neurosci Meth 165 :175–182,2007


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 (*)


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


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


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

150

*

** -50

0

50

100

150

(week) 2 3 4 5 2 3 4 5


Summary

• No effect of mGlu2-PAM in “acute” stress-related paradigms

• mGlu2-PAM reduces dominance level

– Suggestive for antidepressant effect


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.


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


Acknowledgements

• Janssen Neuroscience Discovery and Early Development Teams

• Addex Therapeutics


Documents

Influence of positive allosteric modulation of the mGlu2 ... Donck_310.pdfInfluence of positive allosteric modulation of the mGlu2-receptor on the behavioral responses in animal models