Perospirone, a Novel Antipsychotic Drug, Inhibits Marble-Burying Behavior Via 5-HT1A Receptor in Mice: Implications for Obsessive-Compulsive Disorder

Home , Perospirone

Perospirone, a Novel Antipsychotic Drug, Inhibits Marble-Burying Behavior via 5-HT1A Receptor in Mice: Implications for Obsessive-Compulsive Disorder

Michihiko Matsushita1, Nobuaki Egashira2, Satoko Harada2, Ryoko Okuno2, Kenichi Mishima2, Katsunori Iwasaki2, Ryoji Nishimura1, and Michihiro Fujiwara2,* 1Department of Psychiatry, Fukuoka University School of Medicine, Fukuoka University, 7-45-1, Nanakuma, Jonan-ku, Fukuoka 814-0180, Japan 2Department of Neuropharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Jonan-ku, Fukuoka 814-0180, Japan

Received January 31, 2005; Accepted August 2, 2005

Abstract. Perospirone is a novel atypical antipsychotic drug with dopamine (DA) D2- and serotonin (5-hydroxytryptamine, 5-HT) 5-HT2A-receptor antagonist, and 5-HT1A-receptor agonist properties. In the present study, we examined the effect of perospirone on marble-burying behavior, which has been considered an animal model of obsessive-compulsive disorder (OCD), compared with the effects of other antipsychotics such as haloperidol and risperidone. Perospirone at a dose of 10 mg/kg (p.o.) inhibited marble-burying behavior without affecting the locomotor activity in mice. On the other hand, haloperidol (0.1 mg/kg, i.p.) and risperidone (1 mg/kg, p.o.) showed significant suppression of locomotor activity at the dose that inhibited marble-burying behavior. Furthermore, the inhibition of marble-burying behavior by perospirone was antagonized by WAY100135 (10 mg/kg, i.p.), a selective 5-HT1A-receptor antagonist. WAY100135 at the same dose also antagonized the inhibition of marble-burying behavior by 8- OH-DPAT (3 mg/kg, i.p.), a selective 5-HT1A-receptor agonist. These findings suggest that perospirone may exhibit anti-OCD activity in clinical use and that 5-HT1A-receptor agonistic activity may be involved in the inhibition of marble-burying behavior by perospirone.

Keywords: marble-burying behavior, perospirone, antipsychotic drug, 5-HT1A receptor, obsessive-compulsive disorder

Introduction OCD (4 – 6). In general, marble-burying behavior has been consid- Obsessive-compulsive disorder (OCD) is characterized ered an animal model of OCD (7, 8). In fact, SRI and by recurrent and persistent thoughts, impulses, or images SSRI, which have been used to treat human OCD (obsessions) and/or repetitive, seemingly purposeful symptoms, inhibit marble-burying behavior without behaviors (compulsions) (1). Currently, the serotonin affecting locomotor activity (7 – 9). Moreover, the (5-hydroxytryptamine, 5-HT)-reuptake inhibitor (SRI) inhibition of marble-burying behavior by fluvoxamine, a and selective 5-HT-reuptake inhibitor (SSRI) are SSRI, was antagonized by NAN-190, a 5-HT1A-receptor considered the “first choice” agents for pharmacological antagonist (7). These findings suggest that the 5-HT1A treatment of OCD (2). However, up to 50% of patients receptor may be involved in the inhibition of marble- with OCD fail to respond to a SSRI trial (3). Recently, burying behavior by SSRI. antipsychotics, including haloperidol and risperidone, Perospirone is a novel atypical antipsychotic drug. have reportedly augmented the effect of SSRI therapy in Like other atypical antipsychotics, perospirone antago- nizes the 5-HT2A receptor more potently than the *Corresponding author. FAX: +81-92-863-0389 dopamine (DA) D2 receptor. In addition, perospirone E-mail: [email protected] acts as a 5-HT1A-receptor agonist (10 – 13). The 5-HT1A-

154 Perospirone and Marble-Burying Behavior 155 receptor agonists commonly cause anxiolytic and anti- Drugs depressant effects (14 – 16). Perospirone has also been Haloperidol solution (5 mg/ml, Serenace Injection; reported to exert anxiolytic-like effects in conditioned Dainippon Pharmaceutical Co., Ltd., Osaka) was diluted defensive burying and social interaction tests (17). with saline. Risperidone (Janssen Research Foundation, Based on the unique profile of perospirone’s affinity to Beerse, Belgium), (±)-8-hydroxy-2-(di-n-propyl- the 5-HT1A, 5-HT2A, and D2 receptors, perospirone has amino)tetralin hydrobromide (8-OH-DPAT; Sigma- been postulated to be of possible benefit in treating Aldrich, Inc., St. Louis, MO, USA) and WAY100135 OCD. However, the effect of perospirone on marble- (Tocris Cookson, Inc., Ellisville, MO, USA) were dis- burying behavior has not been studied. In the present solved in saline. Perospirone (Sumitomo Pharmaceutical study, we examined the effect of perospirone on marble- Co., Osaka) was suspended in 0.5% methylcellulose burying behavior compared with the effects of other solution. Risperidone or perospirone was administered antipsychotics such as haloperidol and risperidone. We orally 60 min before the test. Haloperidol, 8-OH-DPAT, also examined the effect of WAY100135 on the inhibi- or WAY100135 was administered intraperitoneally tion of marble-burying behavior by perospirone to (i.p.) 30 min before the test. All drugs were administered investigate the involvement of the 5-HT1A receptor. at a volume of 0.1 ml/10 g of body weight.

Materials and Methods Statistical analyses The values were expressed as means ± S.E.M. The Animals Kruskal-Wallis test followed by the Mann-Whitney U- Five-week old male ICR mice (Nihon SLC Co., test was used for statistical analysis of the data on Shizuoka) were used in each experiment. For at least the number of buried marbles. The results of locomotor seven days before the behavioral tests, the mice were activity were analyzed by a one-way analysis of variance housed in a room under controlled temperature (ANOVA) followed by the Bonferroni/Dunn test. (23 ± 2°C), relative humidity (60 ± 10%), and a cycle of 12 h of light and 12 h of darkness, with the period of Results light starting at 7:00 A.M. Food and water were avail- able ad libitum. The experiments were conducted in Haloperidol at a dose of 0.1 mg/kg (i.p.) significantly compliance with guidelines stipulated by the Animal reduced the number of buried marbles (H(3) = 7.939, Care and Use Committee of Fukuoka University. P<0.05, 0.1 mg/kg: P<0.05, Fig. 1A). At this dose, it also significantly reduced locomotor activity during the Marble-burying behavior test marble-burying behavior (F(3, 30) = 6.834, P<0.01, The marble-burying behavior test is based on the 0.1 mg/kg: P<0.05, Fig. 1B). Risperidone also signifi- method of Ichimaru et al. (7). All experiments were cantly reduced the number of buried marbles at a dose conducted between 10:00 and 17:00. The mice were of 1 mg/kg (p.o.) (H(3) = 11.676, P<0.01, 1 mg/kg: placed individually in clear plastic boxes (30 × 30 × P<0.01, Fig. 2A). At the same dose, it also significantly 28 cm) containing 25 glass marbles (1.5 cm in diameter) reduced locomotor activity during the marble-burying evenly spaced on sawdust 5-cm-deep without food or behavior (F(3, 36) = 6.961, P<0.01, 1 mg/kg: P<0.01, water. At the same time, the locomotor activity of mice Fig. 2B). Conversely, perospirone significantly reduced was measured using an automated activity counter the number of buried marbles without affecting loco- (NS-AS01; Neuroscience, Inc., Tokyo) placed 15-cm motor activity at a dose of 10 mg/kg (p.o.) (H(2) = above the same plastic boxes. This counter has a pyro- 6.518, P<0.05, 10 mg/kg: P<0.01, Fig. 3). Moreover, electric sensor, which can generate a voltage-change by the inhibition of marble-burying behavior by perospirone altering the amount of infrared radiation emitted from was antagonized by WAY100135, a 5-HT1A-receptor the sensor window. The activity was measured with the antagonist (H(2) = 8.823, P<0.05, perospirone + illumination of a 100-W bulb. The results of marble- WAY100135: P<0.05, Fig. 4A). No change in loco- burying behavior were expressed as the number of motor activity during the marble-burying behavior was marbles buried at least two-thirds deep in this paradigm observed at this time (Fig. 4B). Similarly, a 5-HT1A- within 30 min. The average of total counts of locomotor receptor agonist 8-OH-DPAT significantly reduced the activity for 30 min. in the group treated with vehicle was number of buried marbles without affecting locomotor expressed as 100%; values of the group treated with drug activity at a dose of 3 mg/kg (p.o.) (H(3) = 10.001, were expressed as a percentage of variations from values P<0.05, 3 mg/kg: P<0.05, Fig. 5). WAY100135 also of the group treated with vehicle. antagonized the inhibition of marble-burying behavior by 8-OH-DPAT (H(2) = 12.801, P<0.01, 8-OH-DPAT + 156 M Matsushita et al

Fig. 1. Effect of haloperidol on the marble-burying behavior in mice. A: Marble-burying behavior, B: locomotor activity. The values Fig. 2. Effect of risperidone on the marble-burying behavior in ± = are expressed as the means S.E.M. of n 7 – 11 mice per group. mice. A: Marble-burying behavior, B: locomotor activity. The values < *P 0.05, compared to the group treated with vehicle. are expressed as the means ± S.E.M. of n = 8 – 12 mice per group. **P<0.01, compared to the group treated with vehicle.

WAY100135: P<0.01, Fig. 6). In addition, WAY100135 alone at a dose of 10 mg/kg (i.p.) had no effect on the that inhibited marble-burying behavior. Therefore, these number of buried marbles or locomotor activity (data not inhibitions of marble-burying behavior are thought to be shown). due to the suppression of locomotor activity. Perospirone has an agonistic effect on 5-HT1A recep- Discussion tors as well as an antagonistic effect on 5-HT2A and D2 receptors (10 – 13). In the present study, the inhibition In the present study, we demonstrated that perospirone of marble-burying behavior by perospirone was antago- inhibited marble-burying behavior, which has been nized by WAY100135, the selective 5-HT1A-receptor considered an animal model of OCD (7, 8), without antagonist. Therefore, the 5-HT1A-receptor agonistic affecting the locomotor activity in mice. These effects activity may be involved in the inhibition of marble- were therefore not attributable to non-specific sedative burying behavior by perospirone. On the other hand, the effects or a deficit of motor function. Similarly, SRI inhibition of marble-burying behavior by fluvoxamine and SSRI, which have been used to treat human OCD was antagonized by NAN-190, the 5-HT1A-receptor symptoms (2), have also been reported to inhibit marble- antagonist (7). Moreover, the 5-HT1A-receptor agonist burying behavior without affecting locomotor activity 8-OH-DPAT has reportedly inhibited marble-burying (7 – 9). These findings suggest that perospirone, as well behavior without affecting locomotor activity (7). We as SRI and SSRI, may exhibit anti-OCD activity. also found that 8-OH-DPAT inhibited the marble-bury- Conversely, haloperidol and risperidone showed signifi- ing behavior without affecting the locomotor activity, cant suppression of locomotor activity at the dose and this inhibition was clearly antagonized by Perospirone and Marble-Burying Behavior 157

Fig. 3. Effect of perospirone on the marble-burying behavior in mice. A: Marble-burying behavior, B: locomotor activity. The values are expressed as the means ± S.E.M. of n = 8 – 11 mice per group. **P<0.01, compared to the group treated with vehicle. Fig. 4. Effect of WAY100135 on the inhibition of marble-burying behavior by perospirone in mice. A: Marble-burying behavior, B: locomotor activity. The values are expressed as the means ± S.E.M. of n = 8 – 12 mice per group. **P<0.01, compared to the WAY100135. These findings suggest that the 5-HT1A group treated with vehicle; †P<0.05, compared to the group treated receptor may play an important role in marble-burying with perospirone alone. behavior. Several studies suggested that 5-HT2A-receptor antagonistic activity also contributes to the anti-OCD the increases in DA release were greater for a combina- effects. Some drugs with 5-HT2A-receptor antagonistic tion of perospirone and fluoxetine than for a combina- activity potentiated the effect of zimeldine (a SSRI) on tion of risperidone and fluoxetine (18). These findings marble-burying behavior (9). Moreover, the co-admin- may be attributable to the fact that perospirone, unlike istration of risperidone, which exhibits 5-HT2A- and risperidone, has an agonistic effect on 5-HT1A receptors D2-receptor antagonistic activities, with SSRI has been as well as an antagonistic effect on 5-HT2A receptors. reported to be effective in SSRI-refractory OCD patients Based on perospirone’s affinity to not only 5-HT2A (4, 6). However, the pharmacological mechanism receptors but 5-HT1A receptors as well, perospirone responsible for this augmentation remains unclear. The may prove effective in SSRI-refractory OCD patients. combination of risperidone and fluoxetine (a SSRI) has In conclusion, perospirone significantly inhibited been reported to produce remarkable increases in DA marble-burying behavior, which is a model for evaluat- release in the medial prefrontal cortex that are greater ing clinical potential in the treatment of OCD, without than the increases produced by either alone (18). affecting the locomotor activity in mice. Moreover, it Similarly, the combination of perospirone and fluoxetine was thought that 5-HT1A-receptor agonistic activity is markedly increases DA release, and these increases in involved in the inhibition of marble-burying behavior by DA release were suppressed by WAY100635, the perospirone. These findings suggest that perospirone selective 5-HT1A-receptor antagonist (18). Moreover, may exhibit anti-OCD activity in clinical use. 158 M Matsushita et al

Fig. 5. Effect of 8-OH-DPAT on the marble-burying behavior in mice. A: Marble-burying behavior, B: locomotor activity. The values are expressed as the means ± S.E.M. of n = 8 – 12 mice per group. Fig. 6. Effect of WAY100135 on the inhibition of marble-burying *P<0.05, compared to the group treated with vehicle. behavior by 8-OH-DPAT in mice. A: Marble-burying behavior, B: locomotor activity. The values are expressed as the means ± S.E.M. of n = 8 – 9 mice per group. **P<0.01, compared to the group treated with vehicle; ††P<0.01, compared to the group treated Acknowledgments with 8-OH-DPAT alone.

We thank the Janssen Research Foundation (Belgium) and Sumitomo Pharmaceutical Co. (Osaka) for gener- A double-blind, placebo-controlled study of risperidone addition ously donating the risperidone and perospirone used in in serotonin reuptake inhibitor-refractory obsessive-compulsive this study. disorder. Arch Gen Psychiatry. 2000;57:794–801. 5 McDougle CJ, Goodman WK, Leckman JF, Lee NC, Heninger GR, Price LH. Haloperidol addition in fluvoxamine-refractory References obsessive-compulsive disorder. A double-blind, placebo- controlled study in patients with and without tics. Arch Gen 1 Rasmussen SA, Eisen JL. The epidemiology and clinical Psychiatry. 1994;51:302–308. features of obsessive-compulsive disorder. Psychiatr Clin North 6 Sun TF, Lin PY, Wu CK. Risperidone augmentation of specific Am. 1992;15:743–758. serotonin reuptake inhibitors in the treatment of refractory 2 Greist JH, Bandelow B, Hollander E, Marazziti D, Montgomery obsessive-compulsive disorder: report of two cases. Chang SA, Nutt DJ, et al. WCA recommendations for the long-term Gung Med J. 2001;24:587–592. treatment of obsessive-compulsive disorder in adults. CNS 7 Ichimaru Y, Egawa T, Sawa A. 5-HT1A-receptor subtype Spectr. 2003;8:7–16. mediates the effect of fluvoxamine, a selective serotonin 3 Goodman WK, Price LH, Rasmussen SA, Delgado PL, Heninger reuptake inhibitor, on marble-burying behavior in mice. Jpn J GR, Charney DS. Efficacy of fluvoxamine in obsessive-compul- Pharmacol. 1995;68:65–70. sive disorder. A double-blind comparison with placebo. Arch 8 Njung’e K, Handley SL. Evaluation of marble-burying behavior Gen Psychiatry. 1989;46:36–44. as a model of anxiety. Pharmacol Biochem Behav. 1991;38:63– 4 McDougle CJ, Epperson CN, Pelton GH, Wasylink S, Price LH. 67. Perospirone and Marble-Burying Behavior 159

9 Njung’e K, Handley SL. Effects of 5-HT uptake inhibitors, development of novel anxiolytic drugs: models, mechanisms and agonists and antagonists on the burying of harmless objects by future directions. Psychopharmacology (Berl). 1993;112:1–12. mice; a putative test for anxiolytic agents. Br J Pharmacol. 15 Cervo L, Samanin R. Potential antidepressant properties of 8- 1991;104:105–112. hydroxy-2-(di-n-propylamino)tetralin, a selective serotonin1A 10 Hirose A, Kato T, Ohno Y, Shimizu H, Tanaka H, Nakamura M, receptor agonist. Eur J Pharmacol. 1987;144:223–229. et al. Pharmacological actions of SM-9018, a new neuroleptic 16 Kennett GA, Marcou M, Dourish CT, Curzon G. Single admin- drug with both potent 5-hydroxytryptamine2 and dopamine2 istration of 5-HT1A agonists decreases 5-HT1A presynaptic, but antagonistic actions. Jpn J Pharmacol. 1990;53:321–329. not postsynaptic receptor-mediated responses: relationship to 11 Kato T, Hirose A, Ohno Y, Shimizu H, Tanaka H, Nakamura M. antidepressant-like action. Eur J Pharmacol. 1987;138:53–60. Binding profile of SM-9018, a novel antipsychotic candidate. 17 Sakamoto H, Matsumoto K, Ohno Y, Nakamura M. Anxiolytic- Jpn J Pharmacol. 1990;54:478–481. like effects of perospirone, a novel serotonin-2 and dopamine-2 12 Shiwa T, Amano T, Matsubayashi H, Seki T, Sasa M, Sakai N. antagonist (SDA)-type antipsychotic agent. Pharmacol Biochem Perospirone, a novel antipsychotic agent, hyperpolarizes rat Behav. 1998;60:873–878. dorsal raphe neurons via 5-HT1A receptor. J Pharmacol Sci. 18 Yoshino T, Nishijima K, Shioda K, Yui K, Katoh S. 2003;93:114–117. Perospirone, a novel atypical antipsychotic drug, potentiates 13 Tanaka H, Ohno Y, Nakamura M. Localization and pharmaco- fluoxetine-induced increases in dopamine levels via multi- logical characterization of [3H] perospirone-binding sites in rat receptor actions in the rat medial prefrontal cortex. Neurosci brain. Gen Pharmac. 1998;31:159–164. Lett. 2004;364:16–21. 14 Barrett JE, Vanover KE. 5-HT receptors as targets for the