Synthesis and Pharmacological Evaluation of 3-(3,4-Dichlorophenyl)-1-indanamine Derivatives as Nonselective Ligands for Biogenic Amine Transporters

Article abstract of DOI:10.1021/jm0305873

In our efforts toward developing a nonselective ligand that would block the effects of stimulants such as methamphetamine at dopamine (DA), serotonin (5-HT), and norepinephrine (NE) transporters, we synthesized a series of 3-(3,4-dichlorophenyl)-1-indanam

Full text of DOI:10.1021/jm0305873

2624
J . Med. Chem. 2004, 47, 2624-2634
Syn th esis a n d P h a r m a cologica l Eva lu a tion of
3-(3,4-Dich lor op h en yl)-1-in d a n a m in e Der iva tives a s Non selective Liga n d s for
Biogen ic Am in e Tr a n sp or ter s
Han Yu,‡,§ In J ong Kim,‡ J ohn E. Folk,‡ Xinrong Tian,‡,† Richard B. Rothman,| Michael H. Baumann,|
Christina M. Dersch,| J udith L. Flippen-Anderson,#, Damon Parrish,# Arthur E. J acobson,‡ and
Kenner C. Rice*,‡
Laboratory of Medicinal Chemistry, Building 8, Room B1-23, National Institute of Diabetes and Digestive and Kidney
Diseases, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland, 20892-0815,
Clinical Psychopharmacology Section, Intramural Research Program, National Institute on Drug Abuse,
National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland 21224,
and Laboratory for the Structure of Matter, Naval Research Laboratory, Washington, D.C. 20375
Received November 21, 2003
In our efforts toward developing a nonselective ligand that would block the effects of stimulants
such as methamphetamine at dopamine (DA), serotonin (5-HT), and norepinephrine (NE)
transporters, we synthesized a series of 3-(3,4-dichlorophenyl)-1-indanamine derivatives. Two
of the examined higher affinity compounds had a phenolic hydroxyl group enabling preparation
of a medium to long chain carboxylic acid ester that might eventually be useful for a long-
acting depot formulation. The in vitro data indicated that (-)-(1R,3S)-trans-3-(3,4-dichlorophe-
nyl)-6-hydroxy-N-methyl-1-indanamine ((-)-(1R,3S)-11) displays high-affinity binding and
potent inhibition of uptake at all three biogenic amine transporters. In vivo microdialysis
experiments demonstrated that intravenous administration of (-)-(1R,3S)-11 to rats elevated
extracellular DA and 5-HT in the nucleus accumbens in a dose-dependent manner. Pretreating
rats with 0.5 mg/kg (-)-(1R,3S)-11 elevated extracellular DA and 5-HT by approximately 150%
and reduced methamphetamine-induced neurotransmitter release by about 50%. Ex vivo
autoradiography, however, demonstrated that iv administration of (-)-(1R,3S)-11 produced a
dose-dependent, persistent occupation of 5-HT transporter binding sites but not DA transporter
sites.
In tr od u ction
amine-induced subjective effects in humans.6 There are,
presently, no specific treatment agents available for
stimulant abuse, although GBR 12909 is currently being
evaluated as a potential pharmacotherapy for that
purpose7,8 and GBR 12935 has been considered for use
as a treatment agent.
The abuse of methamphetamine and other amphet-
amine-like stimulants has had a disturbing effect on
public health in the United States.1-3 Studies have
shown that methamphetamine can act as a substrate
for transporters of the biogenic amine neurotransmit-
ters, dopamine (DA), serotonin (5-HT), and norepineph-
rine (NE), and can be transported by these transmem-
brane proteins into the nerve terminal.4 The substrate
activity of methamphetamine and related agents evokes
the nonexocytotic release of biogenic amines transmit-
ters. Increased release of DA in the central nervous
system is thought to mediate the dependence-producing
effects of methamphetamine, whereas increased release
of NE in both the peripheral and central nervous system
is thought to induce undesirable cardiovascular side
effects.5 However, recent data suggest that norepineph-
rine may play an important role in mediating amphet-
High-affinity dopamine uptake inhibitors, such as
GBR 12909, selectively block the ability of metham-
phetamine9 to increase extracellular dopamine. We
believe that this effect follows in part from the ability
of GBR 12909 to persistently occupy DA transporter
binding sites.9 However, a limitation of such agents is
that they would not protect against the adverse effects
of the amphetamine-like stimulants that are mediated
by 5-HT and NE transporters. Therefore, we sought to
develop a nonselective transporter ligand that would
bind with high affinity to all three biogenic amine
transporters. This type of medication would ideally
neutralize the neurochemical effects of stimulants such
as methamphetamine and not only attenuate the ad-
dictive effects of these agents but also block cardiovas-
cular and neurotoxic effects.10 Moreover, we wanted this
agent to be amenable to formulation as a long-acting
depot medication. One method that has been used to
prepare an oil-soluble prodrug is to esterify a polar
hydroxyl-containing compound with a medium to long
chain alkanoic acid.9
* To whom correspondence should be addressed. Phone: (301) 496
1856. Fax: (301) 402 0589. E-mail: kr21f@nih.gov.
‡ National Institute of Diabetes and Digestive and Kidney Disease.
† Present address: Procter & Gamble Pharmaceuticals, HCRC,
Mason, OH 45040.
§ Present address: Eli Lilly & Co., Lilly Corporate Center, DC 4813,
Indianapolis, IN 46285.
| National Institute on Drug Abuse.
#
Naval Research Laboratory.
Present address: PDB, Rutgers, the State University of New
J ersey, Piscataway, NJ 08854.
10.1021/jm0305873 This article not subject to U.S. Copyright. Published 2004 by the American Chemical Society
Published on Web 04/14/2004

Indanamine Derivatives
J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 10 2625
Sch em e 1a
a
Reagents and conditions: (a) KOH, EtOH/H2O; (b) TFA, 130 °C; (c) NaBH4, MeOH; (d) SOCl2, 1,4-dioxane; (e) CH3NH2, toluene, 40
°C; (f) diethylamine, toluene, 40 °C; (g) 48% HBr, reflux or BBr3, CH2Cl2, 0 °C.
Indatraline ((()-trans-3-(3,4-dichlorophenyl)-N-meth-
yl-1-indanamine, 1) is an uptake inhibitor with high
cis diastereomer. The alcohol 4 was converted to the
corresponding chloride with a cis/trans ratio of 7:3.
Reacting the chloroindan mixtures with methylamine
or diethylamine in toluene led to the inversion of the
stereocenter to afford the N-methylamine 5 or N,N-
diethylamine 6, respectively, both in cis/trans ratios
of 3:7. The cis and trans isomers of 5 and 6 were
separated and purified by crystallization to give the
racemic cis N-methylamine isomer 7 the cis N,N-
diethylamine isomer 8, the trans N-methylamine isomer
9 and the trans N,N-diethylamine isomer 10. Cleavage
of the methoxy group in the trans amine 9 gave the
phenol 11, and cleavage of the methoxy group in the
trans diethylamine 10 afforded the phenolic compound
12.
affinity for DA, 5-HT, and NE transporters.10,11 It blocks
the transport of methamphetamine into nerve terminals
and therefore blocks the ability of methamphetamine
to release these transmitters.10 Since indatraline (1)
does not contain a hydroxyl group that could serve as
the anchor for the medium to long chain ester function-
ality, we synthesized a series of methoxyl and phenolic
bearing analogues. We previously demonstrated11 that
the biological activity of indatraline is less sensitive to
substitution on the aromatic ring of the indane than
elsewhere in the molecule. Further, a methoxy substitu-
ent on the C6 position of the indane has the least
negative effect on the binding affinity of indatraline
(1).11 Therefore, our synthetic efforts were directed
toward compounds with C6-monosubstituted oxygen-
containing groups.
Optical resolution of 9 was initially accomplished
through diastereomeric salt formation with S-(+)- and
R-(-)-mandelic acids; however, the method was difficult
to replicate. For the reproducible resolution of relatively
small quantities, trans isomer 9 was reacted with (R)-
(-)-1-(1-naphthyl)ethylisocyanate13 to give chromato-
graphically separable diastereomeric ureas (13 and 14,
Scheme 2). The ureas 13 and 14 were converted to (-)-
(1S,3R)-9‚HCl and (+)-(1R,3S)-9‚HCl, respectively, by
transamination.14 The optical purity of (-)-(1S,3R)-9
and (+)-(1R,3S)-9 was determined by 500 MHz NMR
spectra of their diastereomeric ureas 13 and 14 to show
99.8% ee and 97.6% ee, respectively.15
However, to obtain larger quantities of product for
future pharmacological studies in the monkey, we
developed an easily replicable chemical resolution of
racemate 9 using (-)-(R)- and (+)-(S)-O-acetylmandelic
acid (Scheme 3). This gave good yields, and multigram
quantities, initially of a levorotatory R-(-)-O-acetyl-
mandelate salt from which the trans-base (+)-(1S,3R)-
trans-3-(3,4-dichlorophenyl)-6-methoxyl-N-methyl-1-in-
danamine ((+)-(1S,3R)-9) was obtained. A levorotatory
hydrochloride salt was prepared from this (+)-base. The
filtrates from the (-)-(R)-O-acetylmandelate salt were
basified to give a base enriched with (-)-(1R,3S)-trans-
3-(3,4-dichlorophenyl)-6-methoxyl-N-methyl-1-indan-
Ch em istr y
Initial attempts to prepare the needed intermediate,
3-(3,4-dichlorophenyl)-6-methoxylindan-1-one (3), fol-
lowing the procedure of Bogeso12 gave us low yields.
We then attempted to prepare 3 from the 1-(3,4-
dichlorophenyl)-3-(3-methoxylphenyl)-1-propen-3-one (2,
Scheme 1). Cyclizing ketone 2 with AlCl3 or BF3‚OEt
also gave a low yield of 3. However, 3 was obtained in
high yield by heating ketone 2 in neat trifluoroacetic
acid at 120 °C in a sealed tube. Following a modification
of the published procedure,12 3 was first reduced to
3-(3,4-dichlorophenyl)-6-methoxylindan-1-ol (4) as the

2626 J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 10
Yu et al.
Sch em e 2a
a
Reagents and conditions: (a) (R)-(-)-1-(1-naphthyl)ethyl isocyanate, C6H6, room temp; (b) aniline, LiCl, 150 °C.
Sch em e 3a
a
Reagents and conditions: (a) R-(-)-O-acetylmandelic acid, acetone; (b) 20% NH4OH; (c) 48% HBr, reflux; (d) (1) NH4OH, (2) S-(+)-
O-acetylmandelic acid, acetone.
amine ((-)-(1R,3S)-9). Treatment with S-(+)-O-acetyl-
mandelic acid, recrystallization of the dextrorotatory
diastereomeric salt, and subsequent basification gave
multigram quantities of the desired (-)-(1R,3S)-trans-9
as the free base. It was converted to the dextrorotatory
hydrochloride salt. The (-)-(1R,3S)-trans-9 base was
also converted to its ditoluoyl-D-tartaric acid salt for an
X-ray crystallographic study (Figure 1). The optical
purity of both enantiomers of trans amine 9 was found
to be >99% ee, determined as described above in the
chromatographic resolution of 9.
finitive proof of the 1R,3S absolute configuration (Figure
1). The fact that the anion was a D-tartrate salt was
used to determine the absolution configuration of the
cation. The (-)-(1R,3S)-trans-9 and (+)-(1S,3R)-trans-9
were converted to the phenolic compounds (-)-(1R,3S)-
trans-3-(3,4-dichlorophenyl)-6-hydroxy-N-methyl-1-in-
danamine ((-)-(1R,3S)-11) and (+)-(1S,3R)-trans-3-(3,4-
dichlorophenyl)-6-hydroxy-N-methyl-1-indanamine ((+)-
(1S,3R)-11), respectively, with HBr. Compounds 7, 8,
and 10 were not resolved because of their weak affinity
for the norepinephrine transporter (Table 1).
Single-crystal X-ray diffraction analysis of the ditolu-
oyl-D-tartaric acid salt of (-)-(1R,3S)-trans-9 gave de-
We found that in order to prepare a long-chain
decanoate ester, it was necessary to first protect the

Indanamine Derivatives
J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 10 2627
amphetamine-induced (0.5 mg/kg, iv) DA release. As
expected, methamphetamine administration to saline-
pretreated rats produced a rapid and robust increase
in extracellular DA. (-)-(1R,3S)-11 significantly blunted
the peak effect of methamphetamine on dialysate DA
(F[1,20] ) 10.32, P < 0.01). Figure 4 shows the effect of
(-)-(1R,3S)-11 on methamphetamine-induced 5-HT re-
lease in the same rats. In contrast to the DA data, (-)-
(1R,3S)-11 pretreatment did not significantly affect the
peak effect of methamphetamine. A valid interpretation
of the effects of (-)-(1R,3S)-11 pretreatment is compli-
cated by the fact that (-)-(1R,3S)-11 alone increases
dialysate transmitter levels. In an attempt to account
for this problem, the data in Figure 5 show the effects
of (-)-(1R,3S)-11 on methamphetamine-induced trans-
mitter release expressed as a percent of preexisting
baseline (i.e., baseline immediately prior to metham-
phetamine challenge). When the data are expressed in
this manner, (-)-(1R,3S)-11 pretreatment significantly
reduced the peak effect of methamphetamine on the
release of DA (F[1,20] ) 13.13, P <0.001) and 5-HT
(F[1,20] ) 4.27, P <0.05) in vivo.
The ability of (-)-(1R,3S)-11 to persistently occupy
DA and 5-HT transporter binding sites was assessed
using ex vivo autoradiography techniques. Rats were
injected with saline and 1 or 3 mg/kg of (-)-(1R,3S)-11
and were sacrificed 1 h later. Slide-mounted sections
were prepared and labeled with [125I]RTI-55 under
conditions chosen to minimize dissociation of prebound
(-)-(1R,3S)-11. As shown visually in Figure 6, (-)-
(1R,3S)-11 produced a dose-dependent reduction in
5-HT transporter, but not DA transporter, binding sites.
When quantified, the effect on 5-HT transporters was
quite striking, especially compared to the lack of effect
seen with DA transporters (Figure 7).
F igu r e 1. Results of the X-ray analysis on (-)-9 drawn from
the experimentally determined coordinates with the thermal
parameters at the 20% probability level.
secondary amine with a BOC protecting group and then
to react the resulting phenolic compound with 2.8 equiv
of decanoic anhydride to form the decanoate ester.
Removal of the BOC group under mild conditions (CAN,
reflux)16 afforded the decanoate ester 15 (Scheme 4).
Ester 15 was stable for at least a month at room
temperature.
Resu lts a n d Discu ssion
The binding data shown in Table 1 indicate that all
of the compounds have the highest affinity at the 5-HT
transporter, followed by the DA and then the NE
transporter. 3-(3,4-Dichlorophenyl)-6-methoxy-N, N-di-
ethyl-1-indanamines 8, 10, and 12 bind poorly with the
NE transporter. N-Methyl-1-indanamines are generally
potent at all three transporters. As reported,12 racemic
trans amine 9 is more potent than the racemic cis amine
7. Resolution of the trans amine 9 gave the two
enantiomers (+)-(1S,3R)-9 and (-)-(1R,3S)-9. The (-)-
(1S,3R)-9‚HCl is about twice as potent as the racemate
at all of the transporters and considerably more potent
than its enantiomer (+)-(1R,3S)-9‚HCl. This trend was
also observed for the trans phenolic enantiomers (-)-
(1R,3S)-11 and (+)-(1S,3R)-11, derived from the meth-
oxy compounds (-)-(1R,3S)-9 and (+)-(1S,3R)-9, respec-
tively.
On the basis of the fact that there is a significant
difference in the binding affinities of the two enanti-
omers, we next tested the effects of enantiomers (-)-
(1S,3R)-9‚HCl, (+)-(1R,3S)-9‚HCl and (-)-(1R,3S)-11,
(+)-(1S,3R)-11 on the inhibition of DA, 5-HT, and NE
uptake (Table 2). In the methoxy series, the (+)-(1R,3S)-
9‚HCl enantiomer is better able to inhibit the uptake
of neurotransmitters at the DA, 5-HT, and NE trans-
porters, and with the phenolic compounds, the (-)-
(1R,3S)-11 enantiomer was better. In both cases the
enantiomer with the 1R,3S configuration was the more
potent ligand.
Con clu sion s
In our efforts to develop a single molecular entity that
would block the effects of methamphetamine at all three
biogenic amine transporters, we synthesized a series of
3-(3,4-dichlorophenyl)-1-indanamine oxygen-containing
derivatives. All of these compounds had high affinities
at the 5-HT transporter. Their affinities at the NE
transporter, however, were quite different. The relative
and absolute stereochemistry at the C1 and C3 positions
of the indane ring had profound effect on the pharma-
cological activity of the compound. The binding assays
and uptake inhibition data indicated that (-)-(1R,3S)-
trans-3-(3,4-dichlorophenyl)-6-hydroxy-N-methyl-1-in-
danamine ((-)-(1R,3S)-11) had high affinity and high
uptake inhibition potency at all three transporters.
Initial experiments indicated that trans-(-)-(1R,3S)-11
blocked methamphetamine-induced release of DA, 5-HT,
and NE.17 Consistent with this observation, in vivo
microdialysis experiments indicated that trans-(-)-
(1R,3S)-11 inhibited DA and 5-HT transporters, since
iv administration of trans-(-)-(1R,3S)-11 elevated ex-
tracellular DA and 5-HT in a dose-dependent manner.
Although we did not measure extracellular NE in these
experiments, we anticipate that trans-(-)-(1R,3S)-11
will also elevate NE.
The in vivo activity of (-)-(1R,3S)-11 was assessed
using in vivo microdialysis. As shown in Figure 2, iv
administration of (-)-(1R,3S)-11 produced significant
dose-dependent elevations in extracellular DA (F[3,19]
) 8.05, P < 0.001) and 5-HT (F[3,19] ) 7.11, P < 0.002)
in rat nucleus accumbens. The rise in both transmitters
was slow in onset and persistent in duration, even with
iv administration. Figure 3 shows the effects of pre-
treatment with (-)-(1R,3S)-11 (0.5 mg/kg, iv) on meth-
When assessing the effect of putative blocking agents
on methamphetamine-induced neurotransmitter re-
lease, it is important to realize that the uptake inhibitor

2628 J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 10
Yu et al.
Ta ble 1. Binding Affinities of 3-(3,4-Dichlorophenyl)-1-indanamines at the DA, 5-HT, and NE Transporters
Ki ( SD,a nM
5-HTb
26 ( 2
compd
R1
R2
R3
trans or cis
R or S
DAb
NEc
8
CH3
CH3
H
CH3
CH3
CH3
CH3
H
CH2CH3
CH2CH3
CH2CH3
CH2CH3
CH3
cis
racemate
racemate
racemate
racemate
racemate
1R, 3S
1310 70
140 ( 8
40 ( 4
>4700
10
12
7
CH2CH3
trans
trans
cis
1.3 ( 0.1
0.7 ( 0.07
0.3 ( 0.03
1.1 ( 0.08
0.7 ( 0.1
51 ( 4
1010 ( 57
1330 ( 160
170 ( 19
15 ( 0.6
7 ( 0.7
CH2CH3
H
H
H
H
H
H
H
96 ( 2
9
CH3
trans
trans
trans
trans
trans
trans
12 ( 0.3
9 ( 0.3
600 ( 32
11 ( 0.2
12 ( 0.7
86 ( 2
(+)-9‚HCl
(-)-9‚HCl
11
CH3
CH3
1S, 3R
820 ( 35
24 ( 1.4
9 ( 1
CH3
racemate
1R, 3S
3 ( 0.2
(-)-11
H
H
CH3
2 ( 0.3
(+)-11
CH3
1S, 3R
43 ( 4
440 ( 30
indatraline (1)d
1.7 ( 0.2e
0.4 ( 0.04e
6 ( 0.4e
a
b
The Ki values of the test agents were determined in the above assays as described in Biological Methods in ref 16. Dopamine (DA)
and serotonin (5-HT) transporter binding sites labeled with [125I]RTI-55. c Norepinephrine (NE) transporter site labeled with [3H]nisoxetine.
Physical data can be found in ref 10. e Data from ref 9.
d
Sch em e 4a
a
Reagents and conditions: (a) di-tert-butyl dicarbonate, Et3N,
MeOH; (b) decanoic anhydride, NaOH, 2-propanol/water; (c) ceric
ammonium nitrate, MeCN, room temp, then reflux.
Ta ble 2. Reuptake Inhibitory Activities of Indatraline
Analogues at the DA, 5-HT, and NE Transporters
Ki ( SD,a nM
compd
R
R or S
DA
5-HT
NE
F igu r e 2. Effects of (-)-11 on extracellular DA and 5-HT in
rat nucleus accumbens. Rats received iv injections of saline
or (-)-11 at time zero, and dialysate samples were collected
at 20 min intervals for 2 h postinjection. Data are the mean (
SEM for n ) 6 rats/group expressed as % baseline. * ) P <
0.05 compared to saline-injected control rats.
(+)-9‚HCl
(-)-9‚HCl
(-)-11
CH3 1R, 3S 12 ( 0.3 19 ( 1.9 34 ( 1.8
CH3 1S, 3R 600 ( 23 109 ( 4
650 ( 33
20 ( 2
160 ( 12
H
H
1R, 3S 3 ( 0.1
1S, 3R 32 ( 1
2 ( 0.1
41 ( 1
(+)-11
indatraline (1)b
2 ( 0.10c 3 ( 0.16c 11 ( 1.3c
a
The Ki value of the test agents were determined in the above
assays as described in Biological Methods in ref 18. b Physical data
can be found in ref 10. c Data from ref 9.
the baseline observed in the presence of the uptake
inhibitor (peak effect minus blocker baseline, type 2
reduction). On the basis of our experience with GBR
12909 and GBR 12909 decanoate, we expected that
trans-(-)-(1R,3S)-11 would decrease methamphetamine-
induced neurotransmitter release by both measures.
Instead, trans-(-)-(1R,3S)-11 decreased methamphet-
amine-induced DA and 5-HT release only with the
second measurement method (Figure 5). Also, on the
basis of our experience with GBR 12909 and the
itself will elevate extracellular neurotransmitter. The
effect of the uptake inhibitor on methamphetamine-
induced neurotransmitter release can be calculated in
two ways. The first is the reduction in neurotransmitter
release compared to the saline baseline (peak effect
minus saline baseline, type 1 reduction) and second is
the reduction in neurotransmitter release compared to

Indanamine Derivatives
J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 10 2629
F igu r e 3. Effect of pretreatment with (-)-11 (0.5 mg/kg, iv)
on methamphetamine-induced (0.5 mg/kg, iv) DA release. * )
P < 0.05 with respect to saline-pretreated rats.
F igu r e 5. Effect of pretreatment with (-)-11 (0.5 mg/kg, iv)
on methamphetamine-induced (0.5 mg/kg, iv) DA and 5-HT
release when expressed as % change in pre-existing baseline.
* ) P < 0.05 compared to acute saline challenge, and # ) P <
0.05 with respect to saline-pretreated rats.
DA transporters. The modest effect of trans-(-)-(1R,3S)-
11 on methamphetamine-induced neurotransmitter re-
lease and its inability to persistently block DA trans-
porters in the rat suggest that further research will be
needed to determine what properties of reuptake inhibi-
tors predict high efficacy as inhibitors of methamphet-
amine-induced neurotransmitter release.
Viewed collectively, these data indicate that factors
beyond ligand affinity contribute to the ability of an
uptake blocker to attenuate methamphetamine-induced
neurotransmitter release. Although 1R,3S-trans-(-)-11
has high affinity for DA and 5-HT transporters, it
persistently occupies only 5-HT transporters and has
only a modest effect on methamphetamine-induced
neurotransmitter release. This compound will be further
evaluated in monkeys to see whether there are species
differences, and the results will be reported in due
course.
Exp er im en ta l Section
Ch em istr y. Melting points were determined on a MEL-
TEMP II capillary melting-point apparatus and were uncor-
rected. Proton nuclear magnetic resonance (1H NMR) spectra
were recorded in CDCl3 with tetramethylsilane (TMS) as the
internal standard on Varian Gemini-300 and Bruker DMX500
spectrometers. Mass spectra (MS) were recorded on a VG
7070E spectrometer or a Finnigan 4600 spectrometer in the
chemical ionization mode (MS, CI-NH3) and a J EOL SX102a
mass spectrometer in the FAB mode with xenon gas. Optical
rotations were obtained at 20 °C on a Perkin-Elmer 341
polarimeter. Thin-layer chromatography (TLC) was performed
on Analtech silica gel GHLF 0.25 mm plates. Flash column
chromatography was performed with Fluka silica gel 60 (mesh
220-240). Medium-pressure column chromatography (using
silica gel) was carried out at 30 psi with an RT Scientific Co.
(Manchester, NH) high-pressure chromatography system.
F igu r e 4. Effect of pretreatment with (-)-11 (0.5 mg/kg, iv)
on methamphetamine-induced (0.5 mg/kg, iv) 5-HT release.
similarly relatively high affinity of trans-(-)-(1R,3S)-
11 for DA transporters, we anticipated that administra-
tion of trans-(-)-(1R,3S)-11 would persistently occupy
DA transporters. We have speculated, for example, that
the ability of GBR 12909 to persistently occupy DA
transporters contributes to its ability to almost com-
pletely attenuate methamphetamine-induced DA re-
lease. The data (Figures 6 and 7) showed, however, that
trans-(-)-(1R,3S)-11 persistently occupies 5-HT but not

2630 J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 10
Yu et al.
F igu r e 7. Effect of (-)-11 on rat brain biogenic amine
transporters. * ) P < 0.05 compared to saline-injected control
rats.
(()-cis- a n d tr a n s-3-(3,4-Dich lor op h en yl)-6-m eth oxyl-
N-m eth yl-1-in d a n a m in e ((()-7 a n d (()-9). Thionyl chloride
(17.32 g, 145.6 mmol) was added dropwise with stirring to a
solution of (()-4 (30.0 g, 97.0 mol) in dioxane (500 mL) at 0-10
°C. The mixture was warmed to room temperature and was
stirred for 3 h. Then it was quenched with saturated aqueous
NaHCO3, the layers were separated, and the aqueous layer
was extracted with EtOAc (3 × 200 mL). The combined organic
layers were dried (Na2SO4) and evaporated in vacuo to give
an isomeric mixture of the crude cis and trans chlorides in a
7:3 ratio, respectively, that were dissolved in toluene (600 mL).
This mixture was cooled in a dry ice/acetone bath, and
methylamine was passed in until 80.0 g of amine was dissolved
in the solution. The mixture was warmed at 40 °C in a sealed
bottle for 60 h. After removal of the volatile material in vacuo,
the residue was partitioned between H2O and diethyl ether
(400 mL each). The diethyl ether layer was separated and
extracted with 10% aqueous citric acid (400 mL), and the
extract was basified with concentrated NH4OH and extracted
with diethyl ether (400 mL). The diethyl ether was evaporated
to give a mixture of the cis and trans bases ((()-7 and (()-9,
respectively) as a brown oil in a 3:7 ratio. This mixture of bases
(20.0 g, 62.0 mmol) was dissolved in methanol (200 mL), and
oxalic acid (2.79 g, 31 mmol, 1.0 acid equivalent) was added.
After the mixture stood for 2 h at room temperature, the
precipitate was collected and washed with methanol (50 mL).
The solid was partitioned between 10% aqueous NH4OH and
diethyl ether (200 mL each). The aqueous layer was extracted
with diethyl ether (75 mL), and the combined organic layers
were dried (Na2SO4) and evaporated. The resulting oil was
dissolved in acetone (200 mL), and the solution was acidified
with HCl in diethyl ether. The crystalline material that formed
was recrystallized from a mixture of acetone (500 mL) and H2O
(70 mL) to afford 10.3 g (29.6% from (()-4) of the trans salt,
(()-9‚HCl: mp 248 °C; TLC (CH2Cl2/MeOH/concentrated NH4-
F igu r e 6. Ex vivo autoradiographic assessment of the ability
of (-)-11 to persistently occupy DA and 5-HT transporter
binding sites.
Atlantic Microlabs, Inc., Norcross, Georgia, performed elemen-
tal analyses, and the results were within (0.4% of the
theoretical values.
1-(3,4-Dich lor oph en yl)-3-(3-m eth oxylph en yl)-1-pr open -
3-on e (2). By use of a previously described procedure,18,19
2
was prepared11 in 93% yield as a colorless solid from 3,4-
dichlorobenzaldehyde and 3-methoxyacetophenone: mp 116
1
°C; H NMR δ 7.74-7.14 (m, 8H), 3.89 (s, 3H).
3-(3,4-Dich lor op h en yl)-6-m eth oxylin d a n -1-on e ((()-3).
A solution of propenone 2 (34.0 g, 110.0 mmol) in trifluoroacetic
acid (200 mL) was heated at 120 °C for 4 h in a sealed tube.
The trifluoroacetic acid was removed in vacuo, and the
resultant oil was poured into ice/water. The aqueous layer was
separated and extracted with ethyl acetate (400 mL). The
combined organic fractions were neutralized by washing with
saturated NaHCO3 solution. After the fractions were washed
with brine and dried (Na2SO4), the solvent was removed in
vacuo and the product was crystallized from isopropyl ether
to afford 26.1 g (76.8%) of (()-3: mp 81 °C (lit.12 67-69 °C);
1H NMR δ 7.49-6.93 (m, 6H), 4.51-4.47 (m, 1H), 3.88 (s, 3H),
3.30-3.21 (m, 1H), 2.65-2.58 (m, 1H); EIMS m/z 307 (M)+.
3-(3,4-Dich lor op h en yl)-6-m et h oxylin d a n -1-ol ((()-4).
NaBH4 (2.34 g, 62.0 mmol) was added portionwise to a solution
of ketone (()-3 (19.15 g, 62.4 mmol) in MeOH (200 mL) at 0
°C. The reaction mixture was stirred at room temperature for
2 h, and the MeOH was evaporated in vacuo. The resulting
oil was partitioned between diethyl ether and H2O. The aque-
ous layer was extracted with diethyl ether, the combined or-
ganic extracts were dried (Na2SO4), and the solvent was re-
moved in vacuo. Crystallization from a mixture of hexane (40
mL) and EtOAc (40 mL) gave (()-4 as a white solid (14.7 g,
76%): mp 102 °C (lit.12 104-106 °C); 1H NMR δ 7.40-6.83
(m, 6H), 5.26-5.24 (m, 1H), 4.11 (t, J ) 6.2 Hz, 1H), 3.84 (s,
1H), 3.06-2.97 (m, 1H), 1.89-1.83 (m, 1H); CIMS m/z 310
(MH)+.
1
OH, 24:1:0.05) Rf ) 0.45; H NMR δ 7.30-6.71 (m, 6H), 4.18
(t, J ) 7.1 Hz, 1H), 4.06 (t, J ) 6.9 Hz, 1H), 3.75 (s, 1H), 2.96-
2.84 (m, 1H), 2.48 (s, 1H), 1.69-1.55 (m, 1H); CIMS m/z 323
(MH)+. Anal. (C17H17NOCl2‚HCl) C, H, N, Cl.

Indanamine Derivatives
J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 10 2631
The filtrate from the oxalate salt precipitation was evapo-
rated, and the residue was partitioned between aqueous NH4-
OH and diethyl ether. The organic portion was dried (Na2SO4)
and evaporated. The residue was dissolved in acetone, and the
solution was acidified with HCl in diethyl ether. The crystal-
line material was collected after 1 h at room temperature and
recrystallized twice from 95% ethanol/diethyl ether to afford
1.3 g (3.7% from (()-4) of the salt of the cis isomer, (()-7‚HCl:
mp 267 °C (lit.12 262-264 °C); TLC (CH2Cl2/MeOH/concentrated
(d, J ) 8.4 Hz, 1H), 7.11 (d, J ) 2.4 Hz, 1H), 6.94-6.78 (m,
4H), 6.11 (t, J ) 7.5 Hz, 1H), 5.96-5.78 (m, 1H), 4.70 (d, J )
6.9 Hz, 1H), 4.34 (dd, J ) 8.4 and 4.5 Hz, 1H), 3.81 (s, 3H),
2.57 (s, 3H), 2.49-2.28 (m, 2H), 1.71 (d, J ) 6.9 Hz, 3H); MS
(FAB) m/z 519 (M)+. Anal. (C30H28Cl2N2O2) C, H, N, Cl. (-)-
14: mp 88-89 C; [R]20 -117.4 (c 0.89, CHCl3); 1H NMR 8.24
D
(d, J ) 8.4 Hz, 1H), 7.87 (d, J ) 8.4 Hz, 1H), 7.80 (d, J ) 7.5
Hz, 1H), 7.60-7.44 (m, 3H), 7.30 (d, J ) 8.1 Hz, 1H), 7.14 (d,
J ) 2.1 Hz, 1H), 6.93 (d, J ) 7.8 Hz, 1H), 6.87 (dd, J ) 8.4
and 2.1 Hz, 1H), 6.78 (dd, J ) 8.1 and 2.1 Hz, 1H), 6.68 (d, J
) 2.1 Hz, 1H), 6.17 (t, J ) 8.1 Hz, 1H), 5.96-5.86 (m, 1H),
4.72 (d, J ) 7.2 Hz, 1H), 4.40 (dd, J ) 8.7 and 3.9 Hz, 1H),
3.74 (s, 3H), 2.56 (s, 3H), 2.51-2.32 (m, 2H), 1.70 (d, J ) 6.6
Hz, 1H); MS (FAB) m/z 519 (M)+. Anal. (C30H28Cl2N2O2‚
0.5H2O) C, H, N, Cl.
1
NH4OH, 24:1:0.05) Rf ) 0.57; H NMR δ 7.36-6.78 (m, 6H),
4.44 (t, J ) 7.2 Hz, 1H), 4.26-4.23 (m, 1H), 3.82 (s, 3H), 2.51
(s, 3H), 2.49-2.40 (m, 1H), 2.30-2.21 (m, 1H); CIMS m/z 323
(MH)+. Anal. (C17H17NOCl2‚HCl) C, H, N, Cl.
(()-cis- a n d tr a n s-3-(3,4-Dich lor op h en yl)-6-m eth oxyl-
N,N-d ieth yl-1-in d a n a m in e ((()-8 a n d (()-10). Thionyl
chloride (2.60 g, 21.8 mmol) was added to a solution of (()-4
(4.5 g, 14.6 mmol) in dioxane at 0 °C. The mixture was warmed
to room temperature and was stirred for 3 h. After the reaction
was quenched with saturated aqueous NaHCO3, the layers
were separated and the aqueous layer was extracted with
EtOAc three times. The combined organic layers were dried
(Na2SO4) and evaporated in vacuo to give an isomeric mixture
of the crude cis and trans chlorides. The mixture was dissolved
in toluene and diethylamine (8.0 g, 0.11 mol) and was heated
at 40 °C for 60 h. The mixture was cooled and evaporated in
vacuo. The residue was dissolved in diethyl ether, washed with
H2O, and extracted with 10% aqueous citric acid. The extract
was basified with concentrated ammonium hydroxide and
extracted with diethyl ether. The solvent was evaporated to
give a mixture of cis and trans amines in a 3:7 ratio. The
mixture was dissolved in MeOH and was acidified with a
saturated solution of HCl in diethyl ether. The cis amine
hydrochloride ((()-8‚HCl) crystallized as a white solid. It was
recrystallized twice from a mixture of ethanol and diethyl ether
to give (()-8‚HCl (954 mg, 2.63 mmol, 18% yield from (()-4):
mp 200 °C; 1H NMR δ 7.39-6.75 (m, 6H), 4.54 (t, J ) 8.4 Hz,
1H), 4.04 (t, J ) 9.3 Hz, 1H), 3.83 (s, 1H), 2.73-2.39 (m, 5H),
1.88-1.77 (m, 1H), 1.12 (t, J ) 7.2 Hz, 4H); CIMS m/z 365
(MH+). Anal. (C20H23NOCl2‚HCl) C, H, N, Cl. The above filtrate
was concentrated, basified, and extracted with diethyl ether.
The diethyl ether extract was dried, and the solvent was
removed in vacuo. The residue was dissolved in acetone and
acidified with HCl in diethyl ether to give the trans amine
(()-10‚HCl. The hydrochloride salt was recrystallized from a
mixture of acetone and diethyl ether to give pure (()-10‚HCl
(1.75 g, 4.82 mmol, 33% yield from (()-4): mp 131 °C; 1H NMR
δ 7.32-6.77 (m, 6H), 4.67 (t, J ) 6.6 Hz, 1H), 4.33 (dd, J )
3.9 Hz, J ) 13.2 Hz, 1H), 3.83 (s, 1H), 2.62-2.37 (m, 5H), 2.03-
1.95 (m, 1H), 1.06 (t, J ) 7.0 Hz, 4H); CIMS m/z 365 (MH)+.
Anal. (C20H23NOCl2‚HCl) C, H, N, Cl.
(-)-(1R,3S)-tr a n s-3-(3,4-Dich lor op h en yl)-6-m et h oxyl-
N-m eth yl-1-in d a n a m in e ((-)-1R,3S-9) fr om (+)-tr a n s-1-
[3-(3,4-Dich lor op h en yl)-6-m eth oxy-in d a n -1-yl]-1-m eth yl-
3-(1-n aph th alen -2-yl-eth yl)-u r ea ((+)-13), an d (+)-(1S,3R)-
t r a n s-3-(3,4-Dich lor op h e n yl)-6-m e t h oxyl-N -m e t h yl-1-
in d a n a m in e ((+)-(1S,3R)-9) fr om (-)-tr a n s-1-[3-(3,4-
Dich lor op h en yl)-6-m et h oxy-in d a n -1-yl]-1-m et h yl-3-(1-
n a p h th a len -2-yleth yl)u r ea ((-)-14). A mixture of urea (+)-
13 (129.3 mg, 0.248 mmol) and lithium chloride (12.7 mg, 0.3
mmol) in aniline (1.5 mL) was heated at 150 °C for 6 h. After
removal of aniline in vacuo, the residue was diluted with ethyl
acetate and washed with 2 N HCl. The aqueous layer was
extracted with ethyl acetate (2×). The combined organic layer
was washed with H2O and brine, dried over Na2SO4, and
evaporated in vacuo. The residue was purified with column
chromatography (eluted with 3% MeOH in CHCl3 + 0.1%
diethylamine) to give (-)-(1R,3S)-9 (55.2 mg, 69%). A similar
hydrolysis of urea (-)-trans-1-[3-(3,4-dichlorophenyl)-6-meth-
oxyindan-1-yl]-1-methyl-3-(1-naphthalen-2-ylethyl)urea ((-)-
14, 124.6 mg, 0.24 mmol) gave (+)-(1S,3R)-9 (55.0 mg, 71%).
The enantiomeric excess of (-)-(1R,3S)-trans-3-(3,4-dichlo-
rophenyl)-6-methoxyl-N-methyl-1-indanamine ((-)-(1R,3S)-9)-
and (1S,3R)-(+)-trans-3-(3,4-dichlorophenyl)-6-methoxyl-N-
methyl-1-indanamine ((+)-(1S,3R)-9) was determined as
follows.15,20,21 The (-)-(1R,3S)-9 and (+)-(1S,3R)-9 were con-
verted to their diastereomeric ureas (+)-13 and (-)-14, re-
spectively, by reaction with (-)-(R)-1-(1-naphthyl)ethyl isocy-
1
anate. The H NMR 500 MHz spectrum of a 1:1 mixture of 13
and 14 showed the methoxy group at 3.74 and 3.81 ppm in a
ratio of 1:1. The baseline separation allowed quantitation of
the signals. The 1H NMR spectrum of (+)-13 showed two
singlets at 3.74 and 3.81 ppm in a ratio of 99.9:0.1 (99.8% ee),
whereas that of (-)-14 showed two singlets at 3.74 and 3.81
ppm in a ratio of 1.2:98.8 (97.6% ee). The addition of known
amounts of (-)-14 to (+)-13 increased the size of the second
singlet at 3.81 ppm in the spectrum of (+)-13. The size of the
3.74 ppm signal increased, again as expected, when a known
amount of (+)-13 was added to (-)-14.
1R,3S-(-)-tr a n s-3-(3,4-Dich lor op h en yl)-6-m eth oxyl-N-
m eth yl-1-in dan am in e ((-)-(1R,3S)-9) an d (-)-1S,3R-tr a n s-
3-(3,4-Dich lor op h en yl)-6-m et h oxyl-N-m et h yl-1-in d a n -
a m in e ((+)-(1S,3R)-9) fr om Dia ster eom er ic Op tica l Re-
solu tion of (()-tr a n s-3-(3,4-Dich lor op h en yl)-6-m eth oxyl-
N-m eth yl-1-in d a n a m in e ((()-9). A solution of (()-9 (34.33
g, 0.107 mol), prepared from (()-9‚HCl, in acetone (500 mL)
was treated with (-)-(R)-O-acetylmandelic acid (20.68 g, 0.107
mol). Crystallization occurred rapidly upon scratching. After
1 h at room temperature, the crystals were collected, washed
with acetone, and dried to give 24.2 g of fine cotton-like
needles, mp 174-175 °C. Recrystallization was accomplished
by dissolving this solid in a minimum amount of boiling
methanol (ca. 200 mL) and reducing the volume by boiling with
efficient stirring until crystals appeared. At this point acetone
(900 mL) was added and boiling continued until a thick slurry
of needles was obtained (volume ca. 500 mL). This was
repeated with 800 mL of acetone to give a thick slurry of
crystals in ca. 600 mL of acetone. After addition of an
additional 600 mL of acetone, the mixture was brought to room
temperature and allowed to stand for 1 h. The crystals were
(()-tr a n s-3-(3,4-Dich lor op h en yl)-6-h yd r oxyl-N-m eth yl-
1-in d a n a m in e ((()-11). BBr3 was added slowly to a solution
of trans amine (()-9 (1.60 g, 4.98 mmol) in CH2Cl2 (10 mL) at
0 °C. The reaction mixture was stirred at 0 °C for 2 h. The
reaction mixture was poured into a well-stirred mixture of ice
(30 g) and concentrated NH4OH (8 mL), and the two-phase
system was kept at 0 °C overnight. The solid was collected to
give 1.09 g (3.54 mmol, 71% yield) of (()-11. The compound
was recrystallized from EtOH/CH2Cl2 (1:1) to give a white
solid: mp 214 °C (lit.12 206-210 °C); 1H NMR δ 7.36-6.70 (m,
6H), 4.42 (t, J ) 7.8 Hz, 1H), 4.22 (m, 1H), 2.50 (s, 3H), 2.50-
2.41 (m, 1H), 2.27-2.17 (m, 1H); CIMS m/z 309 (MH)+. Anal.
(C16H15NOCl2) C, H, N, Cl.
tr a n s-1-[3-(3,4-Dich lor op h en yl)-6-m eth oxy-in d a n -1-yl]-
1-m eth yl-3-(1-n a p h th a len -2-yl-eth yl)u r ea ((+)-13 a n d (-)-
14). A mixture of racemic 9 (1.84 g, 5.7 mmol) and (R)-(-)-1-
(1-naphthyl)ethyl isocyanate (1.127 g, 5.7 mmol) in benzene
(30 mL) was stirred at room temperature for 1 h. After removal
of solvent, the compounds in the residue was separated with
medium-pressure column chromatography (eluted with 20%
ethyl acetate in hexanes) to give ureas (+)-13 and (-)-14 as
white films in quantitative yield. (+)-13: mp 87-88 C; [R]20
D
+42.0 (c 0.98, CHCl3); 1H NMR 8.20 (d, J ) 8.1 Hz, 1H), 7.89-
7.85 (m, 1H), 7.79 (d, J ) 8.1 Hz, 1H), 7.58-7.43 (m, 3H), 7.30

2632 J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 10
Yu et al.
collected, washed with acetone, and dried to give the R-(-)-
O-acetylmandelate salt of the (+)-(1S,3R)-9 base (18.65 g,
was removed in vacuo, and the product was extracted with
CH2
Cl2 (2 × 3 mL), washed with water (5 mL), and dried (Na2-
67.8%): mp 176-177 °C; [R]20 -53.1° (c 1.1, MeOH). Anal.
SO4). The mixture was concentrated to give an oil that was
chromatographed to give trans-3-(3,4-dichlorophenyl)-6-hy-
droxyl-N-methyl-N-(tert-butoxycarbonyl)-1-indanamine (260
mg, 0.64 mmol, 79% yield) as a clear oil: TLC Rf ) 0.40 (20%
EtOAc/petroleum ether); 1H NMR δ 7.38-6.71 (m, 6H), 6.01-
5.88 (m, 1H), 4.45-4.35 (m, 1H), 2.79 (s, 3H), 2.79-2.41 (m,
1H), 2.38-2.22 (m, 1H), 1.51 (s, 9H); CIMS m/z 410 (MH)+.
D
(C27H27Cl2NO5) C, H, N. The (+)-1S,3R-9 base was obtained
as an oil from its diastereomeric salt using 20% ammonium
hydroxide and diethyl ether extraction, [R]20 +13.4° (c 1.2,
365
MeOH). The (+)-(1S,3R)-9 base from another preparation was
rendered solvent-free by sparging with nitrogen at 60 °C until
constant weight was reached. This material crystallized: mp
45-47 C; [R]20 +13.8° (c 1.365, MeOH). The optical purity
365
To the above amine (260 mg, 0.64 mmol) in 2-propanol (4
mL) was added a solution of NaOH (70 mg, 1.74 mmol) in 1
mL of H2O. Decanoic anhydride (568 mg, 1.74 mmol) was
gradually added, and the reaction mixture was stirred for 30
min at room temperature. Solvent was removed in vacuo, and
the product was extracted with CH2Cl2 (2 × 3 mL), washed
with H2O (5 mL), and dried (Na2SO4). The mixture was
concentrated and chromatographed in a mixture of CH2Cl2 and
MeOH (9:1) to give trans-3-(3,4-dichlorophenyl)-6-decanoate-
N-methyl-N-(tert-butoxycarbonyl)-1-indanamine (311 mg, 0.64
mmol, 87% yield) as a clear oil: TLC Rf ) 0.48 (10% EtOAc/
petroleum ether); 1H NMR δ 7.35-6.89 (m, 6H), 6.03-5.78 (m,
1H), 4.48-4.43 (m, 1H), 2.62 (s, 3H), 2.60-2.42 (m, 3H), 2.39-
2.23 (m, 1H), 1.80-1.69 (m, 2H), 1.58 (s, 9H), 1.57-1.21 (m,
12H), 0.89 (t, J ) 5.9 Hz, 3H); CIMS m/z 565 (MH)+.
To the above amine (311 mg, 0.64 mmol) in 2.5 mL of
acetonitrile was added ammonium cerium nitrate (60 mg, 0.11
mmol). The reaction mixture was stirred at room temperature
for 3 h, then was refluxed at 85 °C for another 3 h. The solvent
was removed in vacuo and the residue was purified by
chromatography to give the deprotected decanoate (()-15 (235
mg, 0.51 mmol, 92% yield) as a clear oil: TLC (50% EtOAc/
petroleum ether) Rf ) 0.42; 1H NMR δ 7.41-6.92 (m, 6H),
4.98-4.88 (m, 1H), 4.68-4.62 (m, 1H), 3.01-2.91 (m, 1H), 2.75
(s, 3H), 2.51 (t, J ) 5.9 Hz, 2H), 2.52-2.37 (m, 1H), 1.75-1.63
(m, 2H), 1.39-1.25 (m, 21H), 0.88 (t, J ) 6.0 Hz, 3H); CIMS
m/z 464 (MH)+.
Biologica l Meth od s. [125I]RTI-55 (0.01 nM, SA ) 2200 Ci/
mmol) was used in the binding assays for the DA (DAT) and
5-HT transporters (SERT).22,23 [3H]nisoxetine (1 nM, SA ) 80
Ci/mmol) was used in the binding assays for the NE transport-
ers (NET).24 All binding assays were performed twice, each in
triplicate, using membranes prepared from frozen rat caudate
for the DAT and SERT assays and frozen whole rat brain
membranes for NET. Briefly, 12 mm × 75 mm polystyrene
test tubes were prefilled with 100 µL of drug, 100 µL of
radioligand, and 50 µL of a blocker or buffer. Drugs and
blockers were made up in 55.2 mM sodium phosphate buffer,
pH 7.4 (with the addition of 5 mM KCl and 300 mM NaCl for
NET binding), containing 1 mg/mL BSA (BB/BSA). Radioli-
gands were made up in a protease inhibitor cocktail containing
BB/BSA, chymostatin (25 µg/mL), leupeptin (25 µg/mL), EDTA
(100 µM), and EGTA (100 µM). Membranes added in 750 µL
of BB initiated the assay. The samples were incubated for 18-
24 h at 4 °C (equilibrium) in a final volume of 1 mL. Brandel
cell harvesters were used to filter the samples over Whatman
GF/B filters, which were presoaked in wash buffer (ice-cold
10 mM Tris-HCl/150 mM NaCl, pH 7.4, containing 2% PEI).
The [3H]DA, [3H]5-HT, and [3H]NE uptake assays were run
as previously reported.25 All uptake assays were performed
twice, each in triplicate, using crude synaptosomes prepared
from rat caudate (for DA) or from rat whole brain minus
caudate (for 5-HT and NE). Briefly, the assays were initiated
by the addition of 100 µL of synaptosomes to 12 mm × 75 mm
polystyrene test tubes, which were prefilled with 750 µL of
[3H]ligand in a Krebs phosphate buffer (pH 7.4) containing 1
mg/mL ascorbic acid and 50 µM pargyline. [3H]NE assays were
performed in the presence of 5 nM 3-(4-iodophenyltropane)-
2-pyrrolidine carboxamide tartrate to prevent reuptake of [3H]-
NE into DA nerves. [3H]5-HT uptake assays were performed
in the presence of 100 nM nomifensine and 100 nM GBR 12935
to prevent reuptake into NE and DA nerves. The incubations
were terminated after 15 min ([3H]DA) or 30 min ([3H]5-HT
and [3H]NE) by adding wash buffer (10 mM Tris-HCl/150 mM
of the (+)-(1S,3R)-9 base, which was determined as above from
the NMR of the urea obtained from reaction with (-)-R-1-(1-
naphthyl)ethyl isocyanate, was >99.4% ee. The (-)-(1S,3R)-
9‚HCl salt was formed upon treatment of the (+)-(1S,3R)-9
base with HCl in diethyl ether. It was recrystallized from
2-propanol isopropyl ether: mp 225-226 °C; [R]20365 -16.4° (c
0.9, MeOH). Anal. (C17H17Cl2NO‚HCl) C, H, N.
The filtrates from the crystallization and recrystallization
of the (+)-(1S,3R)-9‚(-)-(R)-O-acetylmandelate salt were com-
bined, evaporated, and converted to the free base (NH4OH,
diethyl ether) to give material enriched in (-)-(1R,3S)-9 (21.75
g, 0.0675 mol). (+)-(S)-O-acetylmandelic acid (13.1 g, 0.068
mol) was added to the enriched material in acetone. After
standing for 1 h at room temperature, a salt (24.1 g) was
obtained that was recrystallized in a manner similar to its
diastereomer (see above) to give 20.1 g (73.1%) of the (+)-(S)-
O-acetylmandelic acid salt of (-)-(1R,3S)-9: mp 177.5-178.5
C; [R]20 +53.4° (c 0.91, MeOH). Anal. (C27H27Cl2NO5) C, H,
D
N. (-)-(1R,3S)-9 was obtained as an oil from the diastereomeric
salt in a manner similar to its diastereomeric relative (see
above): [R]20 -13.9° (c 1.68, MeOH). The optical purity of
365
the (-)-(1R,3S)-9 base, determined from the NMR of the urea
obtained from reaction with (-)-R-1-(1-naphthyl)ethyl isocy-
anate, was found to be >99.8% ee. The (-)-(1R,3S)-9 base was
converted to its (+)-(1R,3S)‚HCl salt: mp 225-226 °C; [R]20
365
+14.85° (c 0.93, MeOH). Anal. (C17H17Cl2NO‚HCl) C, H, N. The
(-)-(1R,3S)-9 base was also converted to its di-p-toluoyl-D-(+)-
tartrate salt by addition of an equimolar amount of acid to
the base in MeOH. Recrystallization from MeOH afforded the
pure salt: mp 164-166 °C. Anal. (C37H35Cl2NO9‚0.5H2O) C,
H, N.
(-)-(1R,3S)-tr a n s-3-(3,4-Dich lor op h en yl)-6-h yd r oxy-N-
m eth yl-1-in d a n a m in e ((-)-(1R,3S)-11). A mixture of (-)-
(1R,3S)-9 (6.3 g, 19.6 mol) and 48% HBr (85 mL) was heated
at reflux for 45 min. Most of the HBr was removed in vacuo
at 60 °C, the residual oil was dissolved in H2O (150 mL), and
the pH was adjusted to 9.5 with concentrated NH4OH. The
solid formed after standing at 4 °C overnight was recrystallized
from 95% EtOH to give pure (-)-(1R,3S)-11 (2.83 g). An
additional amount of (1R,3S)-(-)-11 was obtained by adjust-
ment of the pH of the mother liquor from the above recrys-
tallization to 9.5 with concentrated NH4OH, followed by
recrystallization from 95% EtOH (total yield 4.81 g, 79.6%):
mp 202-203 °C; [R]20D -19.2° (c 0.9, MeOH); [R]20365 -77.3° (c
0.9, MeOH). Anal. (C16H15Cl2NO) C, H, N.
(+)-(1S,3R)-tr a n s-3-(3,4-Dich lor op h en yl)-6-h yd r oxy-N-
m eth yl-1-in d a n a m in e ((+)-(1S,3R)-11). The (+)-(1S,3R)-11
isomer was prepared in 76% yield in a manner similar to (-)-
(1R,3S)-11 above: mp 200-202 °C; [R]20 +18.3° (c 1.1,
D
MeOH); [R]20365 +76.8° (c 1.1, MeOH). Anal. (C16H15Cl2NO) C,
H, N.
(()-tr a n s-3-(3,4-Dich lor op h en yl)-6-h yd r oxyl-N,N-d i-
eth yl-1-in d a n a m in e ((()-12). The compound was prepared
from amine (()-10 using the procedure described for (()-11:
mp 239 °C; 1H NMR δ 9.74 (s, 1H), 7.61-6.83 (m, 6H), 5.25-
5.22 (m, 1H), 4.59 (t, J ) 8.1 Hz, 1H), 3.32-2.80 (m, 5H), 2.40-
2.29 (m, 1H), 1.31 (t, J ) 6.9 Hz, 3H), 1.22 (t, J ) 6.8 Hz, 3H);
CIMS m/z 351 (MH)+. Anal. (C16H15NOCl2) C, H, N, Cl.
(()-tr a n s-3-(3,4-Dich lor op h en yl)-6-d eca n oa te-N-m eth -
yl-1-in d a n a m in e ((()-15). To a solution of triethylamine (0.5
mL) in methanol (3 mL) at room temperature was added amine
(()-11 (248 mg, 0.81 mmol) in 1 mL of methanol and then di-
tert-butyl dicarbonate (351 mg, 1.61 mmol). The resulting
mixture was stirred at room temperature for 1 h. The solvent

Indanamine Derivatives
J ournal of Medicinal Chemistry, 2004, Vol. 47, No. 10 2633
NaCl, pH, 7.4) and rapidly filtering over Whatman GF/B filters
on a Brandel cell harvester.
crodialysis experiments. The X-ray crystallographic
work was supported in part by NIDA, NIH, DHHS, and
the Office of Naval Research.
In vivo microdialysis studies followed published proce-
dures.26 For autoradiography experiments, rats received injec-
tions of vehicle (n ) 5 animals), 1 mg/kg (-)-(1R,3S)-11 (n )
6 animals), or 3 mg/kg (-)-(1R,3S)-11 (n ) 5 animals) and were
sacrificed 1 h afterward. By use of published methods, 20 µm
coronal sections taken at the caudate level were thaw-mounted
onto subbed glass slides, vacuum-desiccated overnight, and
stored at -80 °C until assayed. Slide-mounted sections were
labeled with [125I]RTI-55 for autoradiographic analysis of
transporters. Slides were thawed at room temperature for 5
min, rinsed for 5 min (10 mM Tris-HCl, pH 7.4, 150 mM NaCl,
1 mg/mL bovine serum albumin, 4 °C), and placed into
cytomailers containing 10 mL of assay buffer, which consisted
of 55.2 mM sodium phosphate buffer (pH 7.4), 1 mg/mL bovine
serum albumin, 1 µg/mL chymostatin, 1 µg/mL leupeptin, 100
nM EDTA, 100 nM EGTA, 1 nM [127I]RTI55, 0.01 nM [125I]-
RTI-55, and appropriate selective transporter blockers. Adja-
cent sections from each brain were incubated in the absence
of blocking drug (total labeling), 100 nM citalopram to select
for DA transporter labeling, 1 µM GBR 12909 to select for 5-HT
transporter labeling, or 10 µM indatraline to determine
nonspecific labeling. After 30 min at 4 °C, slides were rinsed
five times for 1 min (10 mM Tris-HCl, pH 7.4, 150 mM NaCl,
1 mg/mL bovine serum albumin, 4 °C) followed by a 10 s rinse
in deionized water at 4 °C. Slides were vacuum-desiccated
overnight and stored at room temperature until they were
analyzed.
Autoradiographic distribution of 5-HT and DA transporters
was revealed using a Cyclone storage phosphor system (Pack-
ard). Slides were placed into photographic film cassettes and
apposed to storage phosphor screens of the supersensitive
variety. Apposition times were 15 min for DA transporters and
6 h for 5-HT transporters. Transporter levels were quantified
by NIH image software, using [125]iodine microscale reference
standards (Amersham). DA transporter levels were determined
by subtracting nonspecific labeling from labeling in the pres-
ence of 100 nM citalopram. 5-HT transporter levels were
determined by subtracting nonspecific labeling from labeling
in the presence of 1 µM GBR 12909. Results were expressed
as the mean SEM. Statistical significance was determine using
the Students t-test.
Su p p or tin g In for m a tion Ava ila ble: Tables 1-6 of X-ray
crystal data and structural refinement analysis, atomic coor-
dinates, bond lengths and bond angles, anisotropic displace-
ment parameters, hydrogen coordinates, and hydrogen bond
lengths and angles of 1R,3S-(-)-9‚di-p-toluoyl-D-(+)-tartrate.
This material is available free of charge via the Internet at
http://pubs.acs.org.
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