0090-9556/07/3508-1387–1392$20.

00
DRUG METABOLISM AND DISPOSITION Vol. 35, No. 8
Copyright © 2007 by The American Society for Pharmacology and Experimental Therapeutics 15768/3230423
DMD 35:1387–1392, 2007 Printed in U.S.A.

6-Hydroxybuspirone Is a Major Active Metabolite of Buspirone:

Assessment of Pharmacokinetics and 5-Hydroxytryptamine1A
Receptor Occupancy in Rats
Harvey Wong,1 Randy C. Dockens, Lori Pajor, Suresh Yeola, James E. Grace, Jr.,
Arlene D. Stark, Rebecca A. Taub,2 Frank D. Yocca,3 Robert C. Zaczek, and Yu-Wen Li
Departments of Metabolism and Pharmacokinetics (H.W., L.P., J.E.G.), Clinical Discovery (R.C.D.), Biotransformation (S.Y.), and
Neuroscience Biology (A.D.S., R.A.T., F.D.Y., R.C.Z., Y.W.L.), Pharmaceutical Research Institute, Bristol-Myers Squibb,
Wallingford, Connecticut
Received March 16, 2007; accepted May 8, 2007

Downloaded from dmd.aspetjournals.org at ASPET Journals on May 8, 2015

ABSTRACT:

The pharmacokinetics and in vivo potency of 6-hydroxybuspirone respectively. Both compounds appeared to be ⬃4-fold more po-
(6-OH-buspirone), a major metabolite of buspirone, were investi- tent in occupying presynaptic 5-HT1A receptors in the dorsal raphe
gated. The plasma clearance (47.3 ⴞ 3.5 ml/min/kg), volume of than the postsynaptic receptors in the hippocampus. Oral dosing
distribution (2.6 ⴞ 0.3 l/kg), and half-life (1.2 ⴞ 0.2 h) of 6-OH- of buspirone in rats resulted in exposures (area under the concen-
buspirone in rats were similar to those for buspirone. Bioavailabil- tration-time profile) of 6-OH-buspirone and 1-(2-pyrimidinyl)-piper-
ity was higher for 6-OH-buspirone (19%) compared with that for azine (1-PP), another major metabolite of buspirone, that were ⬃12
buspirone (1.4%). After intravenous infusions to steady-state levels (6-OH-buspirone)- and 49 (1-PP)-fold higher than the exposure of
in plasma, 6-OH-buspirone and buspirone increased 5-hydroxy- the parent compound. As a whole, these preclinical data suggest
tryptamine (HT)1A receptor occupancy in a concentration-depen- that 6-OH-buspirone probably contributes to the clinical efficacy of
dent manner with EC50 values of 1.0 ⴞ 0.3 and 0.38 ⴞ 0.06 ␮M in the buspirone as an anxiolytic agent.
dorsal raphe and 4.0 ⴞ 0.6 and 1.5 ⴞ 0.3 ␮M in the hippocampus,

Buspirone (Fig. 1) is a potent and selective 5-HT1A receptor partial humans and rats (Albert et al., 1990). They are abundant as presyn-
agonist that has been prescribed for the treatment of generalized aptic (somatodendritic) autoreceptors on serotonergic neurons, pri-
anxiety disorders (Sramek et al., 2002; Blier and Ward, 2003; Good- marily in the midbrain dorsal raphe nucleus; activation of the 5-HT1A
man, 2004). Although the onset of action of buspirone is relatively autoreceptors inhibits the neuronal activity and results in a reduction
slow and the efficacy is only obtained after chronic treatment, therapy of 5-HT release in terminal synapses of the serotonergic neurons
with buspirone is not associated with undesirable side effects such as (Blier and Ward, 2003). 5-HT1A receptors are also present as postsyn-
sedation, cognitive impairment, withdrawal symptoms, and potential aptic receptors in the forebrain limbic structures; activation of these
abuse liability that can occur with benzodiazepine treatment (Eison receptors inhibits activities of postsynaptic neurons innervated by
and Temple, 1986; Tunnicliff, 1991; Argyropoulos and Nutt, 1999). serotonergic axonal terminals. It has been hypothesized that prolonged
Buspirone has also been suggested to have a role in the treatment of activation of 5-HT1A autoreceptors in the dorsal raphe with 5-HT1A
depressive disorders (Thase et al., 1998; Blier and Ward, 2003).
agonists, such as buspirone and its analogs, results in desensitization
Current hypotheses on the clinical mechanisms of action of buspi-
of the receptors and consequently increases releases of 5-HT in the
rone focus on its agonist activities on 5-HT1A receptors (Schreiber and
limbic regions (Blier and Ward, 2003). The enhanced serotonergic
De Vry, 1993; Blier and Ward, 2003). Belonging to the superfamily
functions are believed to be responsible for anxiolytic and antidepres-
of G-protein-coupled receptors, 5-HT1A receptors share a high iden-
tity (89%) of their transmembrane-spanning amino acid sequences in sant effects of these agents. Although behavioral/clinical effects of
buspirone and other 5-HT1A agonists are believed to be mediated
1
through occupying and acting on 5-HT1A receptors, the level of
Current affiliation: Genentech, Inc., South San Francisco, California.
2 5-HT1A receptor occupancy required for the effects has not been well
Current affiliation: Roche Pharmaceuticals, Nutley, New Jersey.
3
Current affiliation: AstraZeneca Pharmaceuticals, Wilmington, Delaware. investigated. In two human positron emission tomography studies,
Article, publication date, and citation information can be found at little or no occupancy of 5-HT1A receptors has been observed after
http://dmd.aspetjournals.org. administration of clinically efficacious doses of buspirone (Rabiner et
doi:10.1124/dmd.107.015768. al., 2000; Passchier et al., 2001). Preclinically, the requirement of

ABBREVIATIONS: 5HT, 5-hydroxytryptamine; OH, hydroxy; 1-PP, 1-(2-pyrimidinyl)-piperazine; LC/MS/MS, liquid chromatography-tandem mass
spectrometry; WAY-100635, [O-methyl-3H]-N-(2-(4-(2-methoxyphenyl)-1-piperazinyl)ethyl)-N-(2-pyridinyl)cyclohexanecarboxamide trihydrochlo-
ride; AUC, area under the plasma-concentration time profile; EMD 128 130, sarizotan.

1387
1388 WONG ET AL.

FIG. 1. Chemical structure of buspirone

and 6-OH-buspirone.

5-HT1A occupancy for buspirone and its analogs at behaviorally formic acid)] on a Betasil-C18 column (2 ⫻ 100 mm, 5 ␮m) (Thermo Electron
active doses has not been investigated. Corporation, Waltham, MA) at a flow rate of 250 ␮l/min with an analysis time
In humans and rats, buspirone is extensively metabolized and has of 4 min. Detection was performed in positive, multiple reaction monitoring
low oral bioavailability (⬍5%) (Caccia et al., 1983; Jajoo et al., 1989). mode using a Micromass Quattro liquid chromatograph (Waters, Milford, MA)
with an electron ionization source as the LC/MS/MS interface.
The metabolic disposition is similar in the two species with three
In Vivo Autoradiography Studies. Male Sprague-Dawley rats (weighing
major metabolic pathways being N-dealkylation to 1-(2-pyrimidinyl)- 250 –350 g) with dual jugular vein catheterization (Charles River Laboratories,
piperazine (1-PP) and hydroxylation to either 5-hydroxybuspirone or Wilmington, MA) were used in this study. Rats were housed in polycarbonate
6-hydroxybuspirone (Fig. 1). Of these metabolites, 1-PP has been the cages and maintained on a 12:12 h light/dark cycle with free access to standard
most extensively investigated in its role as an active metabolite chow and water. Buspirone (Bristol-Myers Squibb, Wallingford, CT), 6-OH-

Downloaded from dmd.aspetjournals.org at ASPET Journals on May 8, 2015

(Caccia et al.,1986; Zuideveld et al., 2002). 1-PP behaves as an buspirone (Bristol-Myers Squibb), and [3H]WAY-100635 (a selective 5-HT1A
␣2-adrenoceptor antagonist with a low affinity to the 5-HT1A receptor antagonist; GE Healthcare, Chalfont St. Giles, Buckinghamshire, UK) dosing
(Caccia et al., 1986; Gobbi et al., 1991) and therefore is unlikely to solutions were freshly prepared using sterile saline as vehicle. For buspirone
play an important role in the anxiolytic effects of buspirone. Much experiments, rats were injected i.v. with a loading dose of buspirone (0.45–12
less is known about the pharmacological properties of 6-OH-buspi- mg/kg), immediately followed by a continuous infusion of the buspirone at
0.45 to 12 mg/kg/h for 90 min through one jugular vein. For 6-OH-buspirone
rone. Conversion to 6-OH-buspirone has been shown to be the pre-
experiments, rats were injected i.v. with a loading dose of 6-OH-buspirone
dominant metabolic pathway involved in buspirone elimination in (2.4 –23.7 mg/kg), immediately followed by a continuous infusion of 6-OH-
human liver microsomes (Zhu et al., 2005). In addition, plasma levels buspirone at 2.4 to 23.7 mg/kg/h for 90 min through one jugular vein. For both
of 6-OH-buspirone have been recently reported to be 40-fold greater groups of rats, blood samples (0.3– 0.4 ml) were taken at 0, 40, 50, and 60 min
than those of buspirone after oral administration to humans (Dockens postdose through the second jugular vein. In previous pilot experiments, it was
et al., 2006). More recently, 6-OH-buspirone has been found to determined that steady-state plasma concentrations of buspirone and 6-OH-
possess anxiolytic activity in rats using the fear-induced ultrasonic buspirone were achieved by 40 min after the start of the infusion under the
vocalization paradigm (A. D. Stark, unpublished observations). The protocol described (data not shown). Steady-state plasma concentrations (Css)
primary aims of the present study were 1) to evaluate the pharmaco- of buspirone and 6-OH-buspirone are presented as the mean ⫾ S.D. of the 40-,
kinetics of 6-OH-buspirone in rats, 2) to characterize the in vivo 50-, and 60-min time points.
Immediately after the last blood sample, 10 ␮Ci/100 g b.wt. [3H]WAY-
potency of 6-OH-buspirone and buspirone at the 5-HT1A receptor by
100635 (in 0.6 – 0.7 ml of saline) was injected i.v. Rats were decapitated 30
measuring receptor occupancy using in vivo autoradiography, and 3) min later, the brains were collected, frozen, and sectioned (20 ␮m) using a
to investigate the requirement of 5-HT1A occupancy for buspirone at Cryostat, and sections were mounted on Superfrost slides (VWR, Wilmington,
behaviorally active doses. DE). Brain sections were exposed to tritium-sensitive phosphor screens
(PerkinElmer Life and Analytical Sciences, Shelton, CT) for 2 to 3 weeks, and
Materials and Methods images of [3H]WAY-100635 binding in the brain were captured and analyzed
Pharmacokinetic Studies in Rats. Male Sprague-Dawley rats (weighing using a Cyclone Storage Phosphor Imaging System (PerkinElmer Life and
250 –350 g) (Charles River, Wilmington, MA) with single carotid artery Analytical Sciences). The cerebellum, where 5-HT1A receptor density is nom-
catheterization were used in this study. Rats (n ⫽ 3/dose group) received either inal, was used as a reference region for defining nonspecific binding. The
a single 5 mg/kg i.a. dose or a single 10 mg/kg p.o. dose of 6-OH-buspirone percentage occupancy at 5-HT1A receptors in the region of interest was
or buspirone. The vehicle for both the i.a. and p.o. dosing was 0.2 M sodium calculated as 100% ⫺ percent ([3H]WAY-100635 binding in drug-treated ⫺
acetate buffer, pH 4. Serial blood samples (0.25 ml) were collected at predose [3H]WAY-100635 binding in cerebellum)/([3H]WAY-100635 binding in ve-
and 0.1, 0.2, 0.25, 0.5, 1, 2, 4, 6, 8, and 12 h after i.a. dosing and at predose hicle [ [3H]WAY-100635 binding in cerebellum).
and 0.25, 0.5, 0.75, 1, 1.5, 2, 4, 6, 8 and 12 h after p.o. dosing. Immediately Concentrations of buspirone and 6-OH-buspirone in the plasma were quan-
upon collection, the blood was mixed with K3EDTA and stored on ice. Within titated using a LC/MS/MS method. Briefly, 0.1 ml of plasma, 50 ␮l of 200 nM
60 min, blood samples were centrifuged at approximately 1000g for 15 min at internal standard solution, and 0.1 ml of 0.1 M Na2CO3 were mixed followed
4°C, and plasma was harvested. The plasma samples were stored at ⫺70°C by the addition of 1.0 ml of 1:1 methyl tert-butyl ether /ethyl acetate. Samples
until use. were vortexed and centrifuged, and the organic layer was transferred and
Concentrations of buspirone, 6-OH-buspirone, and 1-PP in plasma were evaporated to dryness under nitrogen at 60°C. Residues were reconstituted
quantitated using a liquid chromatography tandem mass spectrometry (LC/ with 0.1 ml of H2O/CH3CN/HCOOH (50:50:0.1, v/v/v). High-performance
MS/MS) method. Briefly, 50 ␮l of plasma, 50 ␮l of 10 ng/ml internal standard liquid chromatography separation was achieved using an acetonitrile (0.1%
solution, and 0.2 ml of phosphate-buffered saline were mixed. Samples were formic acid)/water (0.1% formic acid) gradient on a Zorbax SB-C18 column
passed through a conditioned C18 (EC) solid phase extraction cartridge, (2 ⫻ 50 mm, 5 ␮m) (Agilent Technologies, Palo Alto, CA) at a flow rate of
washed with 1 ml of water and 0.5 ml of 50:50 (v/v) methanol/water, and 200 ␮l/min with an analysis time of 5 min. Detection was performed in
eluted with approximately 2 ml of 3% ammonium hydroxide in acetonitrile. positive, multiple resolution mode using a Micromass Quattro Ultima mass
The eluted sample was transferred and evaporated to dryness under nitrogen at spectrometer (Waters) with an electron ionization source as the LC/MS/MS
40°C. Residues were reconstituted with 0.1 ml of 10:1 (v/v) 100 mM ammo- interface.
nium acetate/ethanol, and 10 ␮l was analyzed using LC/MS/MS. High- Data Analysis. Plots of 5HT1A receptor occupancy versus plasma concen-
performance liquid chromatography separation was achieved using a mobile tration were fitted to a one-site binding model using nonlinear regression
phase consisting of 50% A [aqueous 5 mM ammonium acetate (0.1% formic according to the following equation: % occupancy ⫽ Bmax ⫻ C/(EC50 ⫹ C),
acid)] and 50% B [90:10 methanol/water 5 mM ammonium acetate (0.1% where Bmax is the maximal binding, C is the drug concentration, and EC50 is
 PHARMACOKINETICS AND 5-HT1A RECEPTOR OCCUPANCY 1389
TABLE 1
Pharmacokinetics of 6-OH-buspirone and buspirone in male Sprague-Dawley
rats (n ⫽ 3 per dose route) after single intra-arterial or oral administration
6-OH-Buspirone Buspirone

5 mg/kg 10 mg/kg 5 mg/kg 10 mg/kg

Route i.a. p.o. i.a. p.o.

CL (ml/min/kg) 47.3 ⫾ 3.5 68.7 ⫾ 8.7
Vss (l/kg) 2.6 ⫾ 0.3 2.5 ⫾ 0.3
t1/2 (h) 1.2 ⫾ 0.2 1.2 ⫾ 0.3 0.9 ⫾ 0.4 1.0 ⫾ 0.0
Cmax (␮M) 0.59 ⫾ 0.16 0.04 ⫾ 0.02
tmax (h)a 0.25 (0.25–4.0) 4.0 (0.25–4.0)
F (%) 19.1 ⫾ 9.1 1.4 ⫾ 0.4
CL, plasma clearance; F, oral bioavailability.
a
tmax is presented as median (observed range).

the concentration required for 50% receptor occupancy. Nonlinear regression

was performed using GraphPad Prism (version 3.00; GraphPad Software, San
Diego, CA). Estimates of EC50 are reported as the estimate ⫾ S.E.
All pharmacokinetic parameters were calculated by noncompartmental

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methods as described by Gibaldi and Perrier (1982). Pharmacokinetic param-
eters [aside from the time at which Cmax is observed (tmax)] are reported as the
mean ⫾ S.D. tmax is presented as the median along with the observed range in
parentheses.

Results
The pharmacokinetic parameters of 6-OH-buspirone and buspirone
in rats are summarized in Table 1. Plasma clearances for both buspi-
rone and 6-OH-buspirone were high in relation to liver blood flow in FIG. 2. Plasma concentration-time profile of buspirone and its metabolites, 6-OH-
rats, being 68.7 ⫾ 8.7 and 47.3 ⫾ 3.5 ml/min/kg, respectively (Table buspirone and 1-PP, after i.a. (5 mg/kg) (A) and p.o. (10 mg/kg) (B) administration
1). Estimates of volume of distribution at steady-state (Vss), and of buspirone.
half-life (t1/2) were also were comparable. 6-OH-buspirone had
greater exposure after oral administration than buspirone with higher TABLE 2
estimates of both bioavailability and the maximum observed concen- Exposures of buspirone, 6-OH-buspirone, and 1-PP in male Sprague-Dawley rats
tration achieved after oral dosing (Cmax). (Table 1). Figure 2 shows after single intra-arterial or oral administration of buspirone
plasma concentration-time profiles of buspirone, 6-OH-buspirone, 6-OH-
Buspirone 1-PP
and 1-PP in rats after a single 5 mg/kg i.a. or 10 mg/kg p.o. dose of buspirone

buspirone. Plasma concentrations of both 6-OH-buspirone and 1-PP Buspirone, 5 mg/kg i.a. (n ⫽ 3)
were noticeably higher than those of the parent compound when Cmax (␮M) 5.51 ⫾ 0.65 0.26 ⫾ 0.14 0.69 ⫾ 0.05
AUC (␮M ⴱ h) 3.18 ⫾ 0.38 0.39 ⫾ 0.18 2.24 ⫾ 0.31
buspirone was administered orally. Mean area under the concentra- Buspirone, 10 mg/kg p.o. (n ⫽ 3)
tion-time profile (AUC) estimates for the buspirone metabolites were Cmax (␮M) 0.04 ⫾ 0.02 0.26 ⫾ 0.06 0.69 ⫾ 0.19
approximately 12 (6-OH-buspirone)- and 48 (1-PP)-fold higher than AUC (␮M ⴱ h) 0.09 ⫾ 0.03 1.09 ⫾ 0.21 4.41 ⫾ 0.56
AUC estimates for the parent compound after oral dosing (Fig. 2;
Table 2).
The distribution pattern of 5-HT1A receptors labeled by intravenous In Fig. 4 the relationship between brain 5-HT1A receptor occupancy
injections of [3H]WAY-100635 in the rat brain is consistent with that in hippocampus and dorsal raphe and plasma concentrations of bu-
reported previously (Hume et al., 1994; Khawaja, 1995). A high spirone and 6-OH-buspirone is examined. The data for each brain
density of [3H]WAY-100635 binding appeared in the cortex, septum, region were fitted to a one-site binding model, and the estimated in
hippocampus, hypothalamus, and raphe nuclei in the brainstem. An vivo EC50 values are presented in Table 5. The in vivo affinity of
intravenous infusion of buspirone or 6-OH-buspirone inhibited buspirone (or its metabolites) appeared more potent for 5-HT1A re-
[3H]WAY-100635 in all these regions in a dose-dependent manner. ceptors in the dorsal raphe than in the hippocampus (Table 5; Fig. 4).
Figure 3 shows representative autoradiograms of the inhibitory effect Likewise, 6-OH-buspirone exhibited a higher in vivo affinity in the
of 6-OH-buspirone on in vivo [3H]WAY-100635 binding in the dorsal raphe than in the hippocampus.
forebrain areas including the hippocampus from rats receiving various
doses of 6-OH-buspirone (Fig. 3, A–C). Discussion
The percentage of 5-HT1A receptor occupancy in two representa- The clearance of buspirone in rats estimated from the current study
tive brain regions, the hippocampus and the dorsal raphe nucleus, is similar to the clearance estimate of 51 ml/min/kg observed by
steady-state plasma concentrations, and infusion rates for buspirone Caccia et al. (1983). 6-OH-buspirone showed pharmacokinetics sim-
and 6-OH-buspirone are summarized in Tables 3 and 4. In rats dosed ilar to buspirone in rats with the exception of mean oral bioavailabil-
via an i.v. infusion with buspirone, 6-OH-buspirone Css values were ity, which was ⬃13-fold higher (Table 1). A recent pharmacokinetic
⬃6- to 20-fold less than buspirone Css values. The range of buspirone study in humans reported a similar range of half-lives for both
and 6-OH-buspirone infusion rates resulted in a wide range of steady- buspirone (2.8 – 4.6 h) and 6-OH-buspirone (4.7– 4.3 h) after oral
state concentrations. Occupancy of 5-HT1A receptors increased with administration of buspirone, which is consistent with similar elimina-
increasing steady-state concentrations of both compounds. tion characteristics of both compounds in humans (Dockens et al.,
1390 WONG ET AL.

TABLE 4
Steady-state concentrations and 5-HT1A receptor occupancy in the hippocampus
and dorsal raphe after intravenous infusions of 6-OH-buspirone
5-HT1A Receptor Occupancy
Rat I.D. Infusion Rate Css
Dorsal Raphe Hippocampus

mg/kg/h ␮M %
1 23.7 22.4 ⫾ 2.8 90 85
2 23.7 24.1 ⫾ 1.4 96 98
3 23.7 20.0 ⫾ 2.4 93 93
4 11.9 12.1 ⫾ 0.6 77 75
5 11.9 11.0 ⫾ 2.2 86 77
6 11.9 11.2 ⫾ 0.5 85 78
7 2.4 2.3 ⫾ 0.2 55 35
8 2.4 2.9 ⫾ 0.3 78 52
9 2.4 2.7 ⫾ 0.1 74 44

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FIG. 3. Inhibition of [3H]WAY-100635 binding in rat brain after intravenous infu-
sions of 6-OH-buspirone at various infusion rates. A, vehicle; B, 2.4 mg/kg/h; C,
11.9 mg/kg/h. Hip, hippocampus. Scale bar, 2 mm.

TABLE 3
Steady-state concentrations of buspirone and 6-OH-buspirone, and 5-HT1A
receptor occupancy in the hippocampus and dorsal raphe after intravenous
infusions of buspirone
5-HT1A Receptor
Occupancy
Rat Infusion Buspirone 6-OH-
ID Rate iCss buspirone Css
Dorsal
Hippocampus Raphe

mg/kg/h ␮M ␮M %
A 12.0 11.1 ⫾ 0.1 1.7 ⫾ 0.3 95 100
B 12.0 12.0 ⫾ 0.5 1.6 ⫾ 0.1 85 100
C 4.0 6.36 ⫾ 1.92 0.29 ⫾ 0.04 72 96
D 1.3 1.27 ⫾ 0.09 0.11 ⫾ 0.01 44 87
E 0.45 0.46 ⫾ 0.12 0.04 ⫾ 0.02 28 53
FIG. 4. Relationship between 5-HT1A receptor occupancy in two brain regions
(hippocampus and dorsal raphe) and plasma concentration after intravenous infu-
2006). Currently, there is no literature information on the oral bio- sions of buspirone (A) and 6-OH-buspirone (B).
availability of 6-OH-buspirone in humans.
The in vivo affinity/potency of buspirone and 6-OH-buspirone at TABLE 5
5-HT1A receptors in the brain was examined after intravenous infu- Estimated in vivo EC50 of buspirone and 6-OH-buspirone to 5-HT1A receptors in
sions of both compounds to steady state in the rat. In vivo binding of the hippocampus and dorsal raphe of the rat
[3H]WAY-100635, a selective 5-HT1A antagonist, in the hippocam- In Vivo EC50
pus and dorsal raphe was dose dependently inhibited by buspirone and Compound
Hippocampus Dorsal Raphe
6-OH-buspirone, indicating their interaction with 5-HT1A receptors in
vivo. The in vivo affinity at 5-HT1A receptors for both compounds ␮M
was 3- to 4-fold higher in the dorsal raphe than in the hippocampus Buspirone 1.5 ⫾ 0.3 0.38 ⫾ 0.06
(Table 5). As elucidated in the Introduction, the forebrain regions 6-OH-buspirone 4.0 ⫾ 0.6 1.0 ⫾ 0.3
including the hippocampus express postsynaptic 5-HT1A receptors,
whereas the dorsal raphe posses 5-HT1A somatodendritic autorecep- to underlie the effect of pindolol for augmentation of the selective
tors. A higher affinity to the autoreceptors relative to that for the serotonin reuptake inhibitor antidepressant efficacy (Raurich et al.,
postsynaptic receptors has been observed with pindolol, a ␤-adreno- 1999; Rabiner et al., 2000; Martinez et al., 2001). The dorsal raphe has
receptor antagonist with a high 5-HT1A affinity, and been postulated also been considered to play an important role in the anxiolytic effects
 PHARMACOKINETICS AND 5-HT1A RECEPTOR OCCUPANCY 1391
of 5-HT1A partial agonists, including buspirone, whose effects have after single or multiple clinical doses capable of activating central
been hypothesized to be mediated by desensitizing 5-HT1A autore- 5-HT1A receptor functions (Nakayama et al., 2002; Rabiner et al.,
ceptors in the dorsal raphe (Sim-Selley et al., 2000). Our observation 2002; Bantick et al., 2004). Results of the current study are consistent
of a higher affinity binding of buspirone and its major metabolite, with these literature observations, suggesting that high levels of
6-OH-buspirone, to 5-HT1A autoreceptors in the dorsal raphe supports 5-HT1A receptor occupancy are not required to elicit an anxiolytic
the hypothesis. effect either preclinically in rats and clinically in humans.
Although 6-OH-buspirone was present in the plasma after buspi- In summary, the present study demonstrates that 6-OH-buspi-
rone infusions, the levels of the metabolite were 6- to 20-fold less than rone is the major active metabolite of buspirone with similar in
that of the parent compound. As the in vivo affinity of 6-OH- vivo potency at the 5-HT1A receptor. 6-OH-buspirone has im-
buspirone at the 5-HT1A receptor is comparable to that of buspirone proved oral exposure in comparison with buspirone and could be
(Table 5), the contribution of this metabolite in occupying 5-HT1A an effective anxiolytic agent alternative to buspirone. Finally,
receptors in the buspirone experiments is probably low. Although results of our current study are consistent with literature reports
1-PP was not quantitated in this experiment, it has been shown to have suggesting that a low 5-HT1A receptor occupancy requirement is
low in vitro affinity and selectivity at 5-HT1 receptors in rat brain needed for anxiolytic activity.
(Caccia et al., 1986; Gobbi et al., 1991) and thus is not likely to
contribute significantly to the 5-HT1A occupancy observed in our Acknowledgments. We thank our colleagues Vincenzo Calandra,
studies. Xiao-Xin Yan, Todd Strong, Shelly X. Ren, and Rick Pieschl for
To our knowledge, this is the first published report on the activity contributing to this study.
of 6-OH-buspirone at the 5-HT1A receptor. On the basis of the current

Downloaded from dmd.aspetjournals.org at ASPET Journals on May 8, 2015

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