The Vasopressin V Receptor as a Therapeutic Target in ...€¦ · Psychiatric Drug Discovery and Development Princeton, June 23-24, 2003 Schematic representation of the endocrine,

Psychiatric Drug Discovery and DevelopmentPrinceton, June 23-24, 2003

The Vasopressin V1b Receptoras a Therapeutic Target in Stress-Related Disorders

Guy Griebel


Schematic representation of the endocrine, behavioral and autonomic responses to stress mediated by vasopressin (AVP), and the

consequences of repeated stress

The blockade of V1b receptors in the hypothalamus may prevent the

deleterious effects of an hyperactive HPA (stress) axis


The vasopressin pathways in the brain• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •


Immunohistochemical localization of the V1b receptor in the rat brain

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •


Immunohistochemical localization of V1b receptors in brainareas known to modulate anxiety behaviors in rats

Bed Nucleus of theStria Terminalis

Lateral Septum Amygdala Dendate Gyrus

Control

V1bR


The V1b receptor and stress

Rabadan-Diehl et al., J. Neuroendocrinol. 7 : 903-10, 1995

Immo

V 1b

rece

ptor

mR

NA

(per

cent

of c

ontr

ol)

0

50

100

150

200

250

Control8 days14 days

i.p. HS

**

*

*

Eight or 14 days immobilization stress or hypertonic saline injection (ip HS) increases V1b receptor mRNA


SSR149415 : Chemical Structure

Chemical name : : (2S, 4R)-1-[5-chloro-1-[(2,4-dimethoxyphenyl)sulfonyl]-3-(2-methoxyphenyl)-2-oxo-2,3-dihydro-1H-indol-3-yl]-4-hydroxy-N,N-dimethyl-2-pyrrolidinecarboxamide, isomer(-)

N

NO

O

S

Cl N

O

OH

OO

O

O

C30 H32 Cl N3 O8 SMW = 630.12


Selectivity profile of SSR149415 for vasopressin andoxytocin receptors

SSR149415 is selective for the rat and human V1b receptor

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

Mammary270 ± 39

Kidney2897 ± 509

Liver1050 ± 112

CHO1.3 ± 0.9

Hypophysis3.3

Rat

Itk174 ± 35

CHO1412 ± 214

CHO91 ± 23

CHO1.5 ± 0.8

Hypophysis6.0

Human

OTV2V1aV1bKi (nM)


Efficacy of SSR149415 at the human V1b receptor

Inhibition by SSR149415 of AVP-induced Ca2+

increase in CHO cells transfected with the human V1b receptor.

SSR149415 is a competitive antagonist

10-10 10-9 10-8 10-7

SSR 149415 (M)

0

20

40

60

80

100

IC 50 = 3.8 nMIn

hibi

tion

(% o

f Con

trol

)

ICIC5050 = 3.8 = 3.8 nMnM


• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

Effects of SSR149415 on vasopressin-induced ACTHsecretion in conscious rats

Basal

AC

TH (

pg/m

l)

AVP + SSR149415 (mg/kg p.o., 2 h)

AVP 1 3 10 300

50

100

150

200

250

300

350

AC

TH (p

g/m

l)

**

**

**

AVP + SSR149415 (mg/kg p.o. 2 h)

Basal

SSR149415 decreased in a dose-dependent manner vasopressin-induced secretion of ACTH


Animal models used and psychiatric conditions* modeled to investigate the effects of SSR149415 on emotional processes

• 4-Plate• Light/dark

• Social Interaction• Punished Drinking• Elevated Plus-Maze

Generalized Anxiety Disorder

• Mouse Defense Test Battery

• Forced Swimming• Chronic Mild Stress

• Chronic social stress

PanicDisorder

AcuteStress

Disorder

Major Depressive

Disorder

*According to the DSM-IV classification (1994)

Risk Assessment

Flight Defensive Aggression

• Restraint stress-induced ACTH secretion and

physiological changes• Tail-pinch-induced NE

release• Social Defeat

• Conditioned fear• Distress vocalizations in

rats or guinea pigs

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •


0 10 3 10 30

% T

IME

OPE

N A

RM

S

0

10

20

30

40

50

*

*

mg/kg, po

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

Effects of SSR149415 in two classical models of anxiety: The elevated plus-maze and Vogel conflict tests in rats

SSR149415 produced weak anxiolytic-like activity in the elevated plus-maze and Vogel conflict tests in rats

mg/kg, ip0 .3 1 3 0 1 3 10

NU

MB

ER O

F PU

NIS

HED

R

ESPO

NSE

S

0

9

18

27

36

45

54

* **

* *

Diazepam SSR149415

Elevated Plus-maze Vogel Conflict


mg/kg p.o.0.1 0.3 1 3

TIM

E IN

OPE

N A

RM

S (%

)

0

10

20

30

40

50

60

70

80

90

100

**

$

****

**

$

$$

non stress control stress control stress + SSR149415

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

Effects of SSR149415 in the elevated plus-maze test in mice following social defeat

SSR149415 antagonized the heightened emotionality in the elevated plus-maze produced by prior (stressful) exposure to an aggressive isolated resident


• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

The mouse defense test battery

FLIGHT

RISK ASSESSMENT

DEFENSIVE AGGRESSION


Effects of SSR149415 in the mouse defense test battery

FLIGHT RISKASSESSMENT

DEFENSIVEATTACK

0 1 3 10 0 1 3 10 300

3

6

9

12

**

STO

PS

0 1 3 10 0 1 3 10 300

40

80

120

160

200

**

*

AVO

IDAN

CE D

ISTA

NCE

(cm

)

0 1 3 10 0 1 3 10 300.0

0.5

1.0

1.5

2.0

2.5

3.0

**

*

*

**

BITI

NG

S

SSR149415 reduced defensive aggression, but no other aspects of defensive behaviors

mg/kg, po

Diazepam SSR149415


0 1 10 30

Dur

atio

n (s

)

0

10

20

30

40

50

** **

0 1 10 300

5

10

15

20

25

30

* *

0 1 10 300

2

4

6

8

10

12

14

**

**

Effects of SSR149415 on offensive aggression in hamsters

SSR149415, mg/kg, po

Olfactory Investigation Chase Flank Marking

SSR149415 reduced both conspecific offensive attack and olfactory investigation in hamsters


Effects of SSR149415 on maternal separation-induced distress vocalizations in rat or guinea pig pups

Rat Pups Guinea Pig Pups

SSR149415 produced a dose-dependent decrease in both sonic and ultrasonic distress vocalizations

0 3 10 30 0 3 10 30

ULT

RA

SON

IC D

ISTR

ESS

VOC

ALI

ZATI

ON

S (s

)

0

40

80

120

160

200

240

0 1 3 10 0 10 20 30 SO

NIC

DIS

TRES

SVO

CA

LIZA

TIO

NS

(s)

0

40

80

120

160

200

240

*

**

***

*

* *mg/kg, ip mg/kg, sc

Fluoxetine SSR149415


• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

Effects of SSR149415 on acute stress-induced ACTH or NE secretion in rats

Restraint Stress-induced increase in plasma ACTH levels

Tail Pinch Stress-induced release in NE in the prefrontal cortex

SSR149415 prevented both restraint and tail pinch stress-induced ACTH and NE releases, respectively

ACTH

(pg/

ml)

050

100150200250300350

* *

0 3 10 N

OR

ADR

ENAL

INE

( % b

asel

ine)

100

125

150

175

200

Basal 0 3 10 SSR149415, mg/kg, po SSR149415, mg/kg, ip


• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

Effects of SSR149415 in an animal model of depression: The forced-swimming test in rats

0 10 20 40 0 3 10 30 0 3 10 30

IMM

OB

ILIT

Y T

IME

(se

c)

0

50

100

150

200

250

300

mg/kg, PO

SR149415

***

*

*

*

IMIPRAMINE FLUOXETINE

SSR149415 produced dose-dependent antidepressant-like activity


Chronic sequential appplication of mild stressors*

1 2 3 4 5 6 7 8 9 weeks

Treatments

Tests

•Restraint

•Water and food deprivation

•Paired housing in damp sawdust

•Light/dark cycle modification

•Forced swimming

The Chronic Mild Stress Procedure in Mice :A model of depression

Non-stressedmouse Stressed mouse

*

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •


Effects of repeated treatment (39 days/once a day, ip) of SSR149415 in the chronic mild stress model in mice

Physical state

WEEKS

1 2 3 4 5 6 7

PH

YS

ICA

L S

TATE

SC

ALE

1.0

1.5

2.0

2.5

3.0STRESSED CONTROLSFLUOXETINE (10 mg/kg)SSR149415 (10 mg/kg)SSR149415 (30 mg/kg)

*

CMS CMS+TREAT

**

**

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

Repeated administration of SSR149415 reversed the degradation of the physical state produced by stress


Effects of 39-day treatment (once a day, ip) of SSR149415 in the chronic mild stress model in mice

Anxiety in the Elevated plus-maze

OPE

N A

RM

EN

TRIE

S

0

5

10

15

20

** *

( g g)

*

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

NU

MB

ER O

F O

RIE

NTA

TIO

NS

0

1

2

3

4

5

6

** * *

Risk Assessment in the Mouse Defense Test Battery

Repeated administration of SSR149415 reversed anxiety produced by stress

SSR149415 (10 mg/kg)FLUOXETINE (10 mg/kg)

SSR149415 (30 mg/kg)

NON-STRESSED CONTROLSSTRESSED CONTROLS


No stress

Stress

24h post BrdU

Chronic mild stress decreases the number of BrdU-positive cells

Cell proliferation in the hippocampal dentate gyrus of stressed and non-stressed mice

Num

ber o

f Brd

U p

ositi

ve c

ells

per d

enta

te g

yrus

0

1000

2000

3000

4000

**

no stress stress


Repeated treatment with SSR149415 prevented stress-induced decrease of cell proliferation in the subgranular zone and neurogenesis in the granular cell layer of the dendate gyrus

Effect of SSR149415 on chronic mild stress-induced decrease in neurogenesis in the hippocampus of mice

Num

ber o

f Brd

U p

ositi

ve c

ells

per d

enta

te g

yrus

0

500

1000

1500

2000

t

*

t

no stress stress

vehicleSSR149415Fluoxetine

Num

ber o

f Brd

U p

ositi

ve c

ells

per d

enta

te g

yrus

0

500

1000

1500

2000

2500t

t

*

no stress stress

t

Cell proliferation (24 h post BrdU) Neurogenesis (30 days post BrdU)


Phenotype of BrdU-labeled cells 30 days after the end of stress exposure

Mature neuron

Glial cells 9%

73%

No difference in phenotypic expression patterns between groups

The population of surviving BrdU-positive cells essentially mature into neurons

0

25

50

75

100

Phen

otyp

es o

f Brd

U p

ositi

ve c

ells

(%)

0

25

50

75

100

no stress stress

stress +SSR149415 fluoxetine

NeuNGFAP


The Visible Burrow System: A Realistic Model of Depression in Rats

DOMINANT MALE RAT

FEMALE RATS

SUBORDINATE MALE RATS


Effects of repeated treatment with SSR149415 on agonistic behavior in socially stressed rats in a visible burrow system

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

Fluoxetine and SSR149415-treated animals showed higher wound counts than did controls rats

DAY1 2 3 4 5 6 7 8 9 10 11 12 13 14

NU

MB

ER O

F W

OU

ND

S

0

10

20

30

40

50

60

**

***

SSR149415 (10 mg/kg)FLUOXETINE (10 mg/kg)CONTROLS

ACTH

(pg/

ml)

0

50

100

150

200

DOMINANTSNON-STRESSED CONTROLSSTRESSED CONTROLSFLUOXETINE (10 mg/kg po)SSR149415 (10 mg/kg po)

Fighting intensity with the dominant rat ACTH secretion following restraint stress

SSR149415-treated rats showed much higher plasma ACTH levels relative to vehicle subordinates, suggesting

normalization of this HPA axis parameter


Expected clinical spectrum of therapeutic activity of SSR149415 in anxiety/depressive disorders

Generalized Anxiety Disorder

Panic Disorder Acute Stress Disorder

Major Depressive Disorder

Benzodiazepines

Tricyclics, SSRIs and mixed 5HT/NARIs

SSR149415

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •


SSR149415 : Safety studies

Central depressant effects in mice : rotarod, traction test and spontaneous activity

No effect up to 100 mg/kg, p.o.

Sleep pattern in rats : EEGNo modification up to 30 mg/kg, p.o.

Food intake and weight gain : Obese (ob/ob) andLean female mice, normoglycemic mice and rats

No effect up to 30 mg/kg, p.o.

• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •


Effects of SSR149415 on spatial memory in mice: TheMorris water maze

day1 2 3 4 5

late

ncy

(s)

0

10

20

30

40

50

60

visibleplatform

SALINESSR149415 30mg/kg

DIAZEPAM 3mg/kg

SSR149415 had no effect on either the acquisition of the test or on recalling the platform position after removal.


Effects of SSR149415 in the forced-swimming test in hypophysectomized rats

SSR149415 is still effective in hypophysectomized rats, indicating that the antidepressant-like effects do not depend on blocking only the hypothalamic V1b receptors

0 3 10 30

IMM

OB

ILIT

Y TI

ME

(sec

)

0

20

40

60

80

100

120

SSR149415 (mg/kg, po)

*


Effects of local infusions of SSR149415 in the forced-swimming test in rats

Lateral Septum Central Amygdala

The antidepressant-like effects of SSR149415 are mediated by the V1b receptors located in the lateral septum and the amygdala

Imm

obili

ty ti

me

(s)

0

50

100

150

Saline 0.10.01

**

SSR149415 (µg)

Saline

Imm

obili

ty ti

me

(s)

0

50

100

150

10.10.010.001

***

SSR149415 (µg)


Conclusion• • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • • •

The V1b receptor antagonist SSR149415 is able to attenuate some but not all stress-related behaviors in rodents.

The V1b receptor antagonist showed clear effects only in particularly stressful situations, and in tests sensitive to social or aggression cues.

SSR149415 is devoid of central depressant effects, even at high doses, and does not affect cognitive processes or food intake, suggesting a large therapeutic window.

The lateral septum and the central nucleus of the amygdala participate in the antidepressant-like action of SSR149415

V1b receptor antagonists might be useful as a treatment for major depression and stress disorders that result from traumatic events


CHEMISTRY

ELECTROPHYSIOLOGICAL STUDIES

P. AVENETM. DECOBERTD. FRANCON

BEHAVIORAL STUDIES

NEUROCHEMICAL STUDIES

M. ARNONEO. BERGISD.C. BLANCHARDR.J. BLANCHARDE. DUCONSEILLEJ. SIMIANDP. SOUBRIEJ. STEMMELIN

R. ALONSOL. LUKOVICC. SERRADEIL-LE GALR. STEINBERG

Acknowledgments

J. WAGNON

Documents

The Vasopressin V Receptor as a Therapeutic Target in ...€¦ · Psychiatric Drug Discovery and Development Princeton, June 23-24, 2003 Schematic representation of the endocrine,