'Click' D1 receptor agonists with a 5-HT1A receptor pharmacophore producing D2 receptor activity

Article abstract of DOI:10.1016/j.bmc.2009.06.019

A series of new 1-aryl-3-benzazepine derivatives containing an arylpiperazinyl function as the N3 substituent were synthesized by combining a D₁ receptor agonistic pharmacophore and a 5-HT_1A receptor pharmacophore through Click reaction. Interestingly, these compounds generally do not have good binding affinity at the D₁ receptor, but most compounds are potent at both D₂ and 5-HT_1A receptors. Compound 8h, containing 1-m-tolyl-benzazepine scaffold and 2-methoxyphenylpiperazine core, displayed good affinity at all tested receptors, with K_i values of 144, 80, and 133 nM, for the D₁, D₂, and 5-HT_1A receptors, respectively. Compound 13 with the triazole moiety formed differently from that in 8h showed the highest affinity at the D₂ receptor with K_i value of 19 nM. This compound also showed moderate affinity at the 5-HT_1A (K_i, 105 nM), and D₁ (K_i, 551 nM) receptors. Functional assays indicated that both compounds 13 and 8h are antagonists at D₁ and D₂ receptors, whereas full agonistic activity at the 5-HT_1A receptor was observed. In agreement with the binding affinity, compound 13 is a high efficacy D₂ antagonist and 5-HT_1A agonist.

Full text of DOI:10.1016/j.bmc.2009.06.019

Bioorganic & Medicinal Chemistry 17 (2009) 4873–4880
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
‘Click’ D1 receptor agonists with a 5-HT1A receptor pharmacophore producing
D2 receptor activity q
a,
Jing Zhang a, , Hai Zhang b, , Wenxian Cai b, Leiping Yu b, Xuechu Zhen b, , Ao Zhang
*
*
a Synthetic Organic and Medicinal Chemistry Laboratory (SOMCL), Shanghai Institute of Materia Medica (SIMM), Chinese Academy of Sciences, Shanghai 201203, China
b Neuropharmacological Laboratory, Shanghai Institute of Materia Medica (SIMM), Chinese Academy of Sciences, Shanghai 201203, China
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of new 1-aryl-3-benzazepine derivatives containing an arylpiperazinyl function as the N3 substi-
tuent were synthesized by combining a D1 receptor agonistic pharmacophore and a 5-HT1A receptor
pharmacophore through Click reaction. Interestingly, these compounds generally do not have good bind-
ing affinity at the D1 receptor, but most compounds are potent at both D2 and 5-HT1A receptors. Com-
pound 8h, containing 1-m-tolyl-benzazepine scaffold and 2-methoxyphenylpiperazine core, displayed
good affinity at all tested receptors, with Ki values of 144, 80, and 133 nM, for the D1, D2, and 5-HT1A
receptors, respectively. Compound 13 with the triazole moiety formed differently from that in 8h showed
the highest affinity at the D2 receptor with Ki value of 19 nM. This compound also showed moderate affin-
ity at the 5-HT1A (Ki, 105 nM), and D1 (Ki, 551 nM) receptors. Functional assays indicated that both com-
pounds 13 and 8h are antagonists at D1 and D2 receptors, whereas full agonistic activity at the 5-HT1A
receptor was observed. In agreement with the binding affinity, compound 13 is a high efficacy D2 antag-
onist and 5-HT1A agonist.
Received 27 May 2009
Revised 10 June 2009
Accepted 11 June 2009
Available online 16 June 2009
Keywords:
Arylbenzazepine
Arylpiperazine
Click reaction
Dopamine receptor
Serotonin 5-HT1A receptor
Ó 2009 Elsevier Ltd. All rights reserved.
1. Introduction
optimal option for treating these disorders.14–17 In fact, sarizotan,
a compound possessing high binding at both D2 and 5-HT1A recep-
Dopamine (DA) D1 and D2 receptors represent the most abun-
dant DA receptors in the mammalian brains, and are implicated
in the pathophysiology of several neurobehavioral disorders, such
as Parkinson’s disease, and a number of other movement and
hyperactivity disorders, including schizophrenia, mania, depres-
sion, substance abuse, and eating disorders.1,2 Indeed, D1 and D2
receptors are the most studied therapeutic targets for these neuro-
logical and psychiatric diseases, and compounds targeting these
two receptors generally have utilities in the treatment of Parkin-
son’s disease, depression, and schizophrenia.3
Recently, accumulating evidences have indicated that serotonin
5-HT1A receptor is also implicated in many CNS disorders, includ-
ing anxiety,4 depression,5 neuroprotection,6,7 schizophrenia,8,9 Par-
kinson’s disease,10–12 and Alzheimer disease13 by acting alone or
together with other neurotransmitter receptors, especially DA
receptor. Therefore, it has been proposed that 5-HT1A agonists
combined with DA receptor agonism or antagonism may be an
tors, and acting as a 5-HT1A agonist and D2 receptor partial agonist/
antagonist, has been found in effectively improving DA-induced
motor complications by reducing striatal serotoninergic nerve im-
pulse activity without altering
have been in clinical trial as an innovative bisfunctional drug for
treating dyskinesias associated with
-dopa therapy in PD.18
L-dopa efficacy. These compounds
L
1-Aryl-3-benzazepines, including SKF-38393 (1) and SKF-83959
(2) (Fig. 1), represent the prototypical structural scaffold possessing
D1 receptor activities.19–22 These compounds generally display
high binding affinity and selectivity at the D1 receptor, and are use-
ful tool drugs for the study of the receptor and its therapeutic indi-
cations. However, poor intrinsic activity, low metabolic stability,
and several unwanted side effects are generally associated with
these compounds.19 We recently found that compounds (e.g., 3)
with a larger lipophilic substituent at the C6 position displayed
equi-potent or even higher binding affinity than the C6 non-substi-
tuted prototypes.20,21 As a continuation of this work toward a full
understanding on the SAR of this scaffold, we decided to explore
the relatively not-well-explored N3 position of the benzazepine
template where only a few N-substituents including H, Me, allyl,
propargyl, and few alkylamino groups have been reported.19
Our original objective is to incorporate a 5-HT1A agonistic phar-
macophore as the N3 side chain in the 1-aryl-3-benzazepine scaf-
fold to achieve compounds possessing both D1 and 5-HT1A
agonistic properties.21 Since 1-arylpiperazin-4-yl functionality is
q
This work has been partially presented at the 237th American Chemical Society
National Meeting at Salt Lake City, UT 2009, MEDI 0311.
* Corresponding authors. Tel./fax: +86 21 50806750 (X.Z.); tel.: +86 21 50806035;
fax: +86 21 50806600 (A.Z.).
E-mail addresses: xzhen@mail.shcnc.ac.cn (X. Zhen), aozhang@mail.shcnc.ac.cn
(A. Zhang).
These two authors contributed equally to this work.
0968-0896/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2009.06.019

4874
J. Zhang et al. / Bioorg. Med. Chem. 17 (2009) 4873–4880
Cl
azidation with NaN3 in 70–75% overall yield. Treating 5 and 6 by
Me
N
6
Click reaction28 under CuI/DIPEA provided triazole 7 in 82–99%
yields. Compounds 7 were O-demethylated using BBr3 (1 M, in
CH2Cl2) at ꢀ78 °C yielding the final compounds 8a–h in 17–70%
yields. The general low yields were due to the incomplete demeth-
ylation during this reaction which led to monohydroxy products.
Similarly, benzazepine 4 (R = 20-Me) was treated with chloropropyl
bromide to yield the corresponding chloride 9 in 80% yield
(Scheme 1). Azidation of chloride 9 with NaN3 provided azide 10
in 90% yield. Reaction of the corresponding arylpiperazine with
pent-4-ynyl 4-methylbenzenesulfonate afforded compound 11 in
76% yield. Click reaction of azide 10 and alkyne 11 yielded the dim-
ethoxybenzazepine 12 in 97% yield, which was then treated with
BBr3 (1 M, in CH2Cl2) at ꢀ78 °C providing the final compound 13
(R = 20-Me, n = m = 1, Ar = 2-MeO-Ph). However, this compound
was not stable at storage, and a side compound with very similar
polarity on TLC was observed after two days. After several trials
on chromatography and preparative TLC, we were unable to isolate
the side compound in pure form, therefore no other analogues
were made in this series.
HO
HO
HO
HO
3N-Me
NH
1
OH
HO
1, (+)-SKF-38393
3
2, (+)-SKF-83959
HO
HO
N
N
N
n
m
N
N
N
R
R'
5-HT1A pharmacophore
Proposed 1-aryl-3-benzazepines
"Click" reaction
D1 pharmacophore
Figure 1. Representative and proposed 1-aryl-3-benzazepines.
a well-established pharmacophore for the 5-HT1A receptor,23–25 it
was selected to attach to the N3 position of 1-aryl-3-benzazepines
through a linker by a Click reaction (Fig. 1). To our surprise, all
these new compounds significantly lost binding affinity at the D1
receptor, and instead high affinity at the D2 receptor, along with
good 5-HT1A binding, was observed. Herein, in this report, we de-
scribe the details of the synthesis and pharmacological investiga-
tions of these compounds.
The control compound 16 was prepared in a similar manner.
Treatment of diethylamine with tosylate 14 which was prepared
from pent-4-yn-1-ol provided N,N-diethylpent-4-yn-1-amine 15
in quantitative yield. Click reaction of 15 with azide 6 in the pres-
ence of CuI/DIEPA in THF at RT provided triazole 16 as a colorless
liquid in 90% yield (Scheme 2).
2. Chemistry
3. Results and discussion
20-Methyl or 30-methyl substituted benzazepines 4 were pre-
pared from 3,4-dimethoxyphenylethylamine and corresponding
styrene oxide by using a literature procedure.20,26,27 Treating 427
with pentyn-4-yl 4-methylbenzenesulfonate which was prepared
by tosylation of the corresponding alcohol provided N-alkylated
benzazepine 5 in 60–80% yield. Azide 6 was prepared from the cor-
responding arylpiperazine and chloroalkyl bromide followed by
All the new compounds (8a–h, and 13, 16) were racemic, and
were converted to their HBr or TFA salts for the bioassay. The bind-
ing affinity of these compounds was assayed at D1, D2, and 5-HT1A
receptors using membrane preparation obtained from stable
transfected HEK293 or CHO cells (Table 1). This procedure is sim-
ilar to those reported by us previously.15,19,20 [3H]SCH23390,
MeO
MeO
MeO
MeO
MeO
NH
R
N
R
Cl
N
R
N3
Br
Cl
d
n
n
n
MeO
c
4
9
10
a
a
MeO
MeO
N
Ar
N
Ar
N
NH
m
N
R
11
n
b
c, d
N
Ar
N
N3
m
5
6
b
MeO
MeO
MeO
Ar MeO
N
N
N
m
N
n
N
m
N
R
N
N
n
N
N
N
N
Ar
R
e
7
12
13
e
HO
HO
HO
HO
N
m
N
N
N
N
N
m
N
n
n
N
N
N
N
N
Ar
Ar
R
R
(R = 2'-Me, n = m = 1, Ar = 2-MeO-Ph)
8
a, R = 2'-Me, n = m = 1, Ar = 2-Pym
b, R = 2'-Me, n = m = 1, Ar = 2-MeO-Ph
c, R = 2'-Me, n = m = 1, Ar = 2-OH-Ph
d, R = 3'-Me, n = m = 1, Ar = 2-Pym
e, R = 3'-Me, n = m = 1, Ar = 2-MeO-Ph
f, R = 3'-Me, n = m = 1, Ar = 3-BIT
g, R = 3'-Me, n = 0, m = 1, Ar = 2-MeO-Ph
h, R = 3'-Me, n = 2, m = 1, Ar = 2-MeO-Ph
N
S
N
N
Pym
BIT
Scheme 1. Synthesis of compounds 8a–h, and 13 via Click reaction as the key step. Reagents and conditions: (a) alkynyl tosylate, K2CO3, MeCN, reflux; (b) CuI, DIPEA, THF, rt,
overnight; (c) chloroalkyl bromide, K2CO3, MeCN, 50 °C; (d) DMSO, NaN3, 100 °C; (e) BBr3, CH2Cl2.

J. Zhang et al. / Bioorg. Med. Chem. 17 (2009) 4873–4880
4875
a
the benzo[d]isothiazol-3-yl moiety (BIT) provided compound 8f
which is equi-potent at all the three tested receptors (D1, D2, 5-
HT1A) with Ki values of 221, 200, and 235 nM, respectively. The
length of the linker between the benzazepine core and the triazole
moiety played a remarkable influence on the binding at the D2
receptor, but only a minor influence on binding affinity at the D1
and 5-HT1A receptors. Compared to 8e, compound 8g with a short-
er linkage lost binding affinity at the D2 receptor; however, com-
pound 8h containing a longer chain displayed an enhancement
for both D1 and D2 receptors, and the affinity at the 5-HT1A receptor
was well retained. Therefore, compound 8h has good affinity at all
the tested receptors (D1, D2, 5-HT1A) with Ki values of 144, 80, and
133 nM, respectively. Interestingly, compound 13 with the triazole
moiety formed differently from that in 8a–h showed the highest
affinity at the D2 receptor with Ki value of 19 nM. This compound
also showed moderate affinity at the 5-HT1A receptor (Ki,
105 nM), along with moderate affinity at the D1 receptor (Ki,
551 nM). Therefore, compound 13 is D2-selective binder and is
5.5- and 29-fold more potent at this receptor against the 5-HT1A
and D1 receptor, respectively.
OTs
N
NEt2
15
14
MeO
OCH3
N
N
N
N
N3
N
Et2N
N
b
16
6
Scheme 2. Synthesis of control compound 16. Reagents and conditions: (a)
diethylamine, K2CO3, MeCN, reflux; (b) CuI, DIPEA, THF, rt, overnight.
Table 1
In vitro binding assays of new compounds 8a–h, 13, and 16 at D1, D2, and 5-HT1A
receptors in HEK293 or CHO cellsa
Compounda
Ki (nM)
D1 ([3H]SCH23390)
D2 ([3H]Spiperone)
5-HT1A ([3H]8-
OH-DPAT)
1, ( )-SKF-38393
2 ( )-SKF-83959
393 5 (190b)
1.93 0.47 (1.18c)
NA
NA (720,000b)
NA
NA
>1000
8a
8b
8c
8d
8e
8f
8g
8h
13
16
NA
799 37
85 15
98 11
648 83
133 5.4
235 78
457 87
133 6.2
105 28
381 92
Since all the final compounds (8a–h, 13) only contain pharma-
cophores for the D1 (benzazepine core) and 5-HT1A (arylpiperazine
core) receptors, it is intriguing that where the extra D2 binding
affinity observed from most of these compounds is originated. To
examine the possible role of the triazole-containing linker,
compound 16 which does not contain the benzazepine core was
prepared as a control and was evaluated in the same conditions.
To our surprise, this compound is indeed inactive at both D1 and
D2 receptors, except its moderate 5-HT1A receptor binding (Ki,
381 nM). Therefore, it is likely that substitution on the N3 of ben-
zazepine scaffold produced the observed D2 receptor affinity.
Compound 13, which is the most potent at the D2 receptor, and
compound 8h, which has good affinity at all the three tested recep-
501 74
NA
39.1 10.5
NA
963 171
655 0.4
221 29
492 165
144 50
551 0.1
NA
NA
115 20
200 35
NA
80.0 10.3
19.1 1.3
NA
a
Values are means of three to five experiments, and all compounds were tested
as HBr or THF salts in racemates, NA = not active (less than 80% of inhibition in
radioligand binding at 10
l
M), NT = not tested.
b
Data from Ref. 26.
Data from Ref. 22.
c
tors, were further evaluated in the [35S]GTP
cS binding assays to
determine their agonistic and antagonistic activities. The results
are summarized in Figure 2 and Table 2.
[3H]Spiperone, and [3H]8-OH-DPAT were used as the standard
radioligands for DA D1, D2, and serotonin 5-HT1A receptors, respec-
tively. Racemic ( )-SKF-38393 (1) and ( )-SKF-83959 (2) were also
tested in our assays for comparison.
As shown in Figure 2 and Table 2, both compounds 8h and 13
did not show appreciable stimulation at both D1 and D2 receptors
(Fig. 2 a and b) in the [35S]GTP
cS binding assays indicating that
Compound 8a which contains an ortho-tolyl as the C1 substitu-
ent in the benzazepine core and pyrimidin-2-yl (Pym) as the aryl
group did not show binding affinity at both D1 and D2 receptors;
however, some affinity (Ki, 800 nM) was observed at the 5-HT1A
receptor. Modification of the aryl fragment in the arylpiperazine
core provided compounds 8b,c. Compound 8b containing 2-MeO-
Ph as the aryl group displayed significant binding affinity at both
D2 and 5-HT1A receptors with Ki values of 39 and 85 nM, while
moderate affinity (Ki, 501 nM) was obtained at the D1 receptor.
This compound can be viewed as a bisfunctional compound for
D2 and 5-HT1A receptors, with a twofold binding selectivity for
the D2 receptor. Removal of the methyl group in the 2-MeO-Ph
fragment yielded compound 8c which was inactive at both D1
and D2 receptors. Interestingly, compound 8c retained good affin-
ity at the 5-HT1A receptor (Ki, 98 nM). This result indicated that
the aryl group in the arylpiperazine function is not only a determi-
nant factor for the 5-HT1A receptor, but also a crucial factor for both
D1 and D2 receptors.
they lack agonistic activities at the two DA receptors. However,
co-administration of these compounds with their corresponding
standard full agonists (SKF 38393 for the D1, and quinpirole for
the D2 receptors), these two compounds displayed a strong inhibi-
tion to the binding of the standard agonists at the corresponding
receptors. Therefore, compounds 8h and 13 are antagonists at both
D1 and D2 receptors. In relevance to their binding affinity, com-
pound 13 has stronger antagonistic activity than compound 8h,
especially at the D2 receptor with an IC50 value of 480 nM
(1.57 lM for 8h). Different from the functional profiles at the D1
and D2 receptors, compounds 8h and 13 displayed strong agonistic
activity (Emax >100%) at the 5-HT1A receptor (Table 2, Fig. 2 c),
which is even higher than the standard agonist, 5-HT (Emax = 100%).
These compounds are full 5-HT1A agonist with EC50 values of 200
and 330 nM, respectively.
4. Conclusion
Among the C1 meta-tolyl substituted benzazepine derivatives
8d–h, the Pym containing compound 8d showed moderate affinity
at both D1 and 5-HT1A receptors, and is inactive at the D2 receptor.
Again, replacing the Pym function with 2-MeO-Ph yielded com-
pound 8e showing a significant enhancement in binding to the
D2 receptor, together with a moderate increase in binding to the
D1 and 5-HT1A receptors. This compound is equipotent at both D2
and 5-HT1A (Ki, 115 and 133 nM, respectively), and is fivefold more
potent against the D1 receptor. Further changing the 2-MeO-Ph to
In summary,
a series of new benzazepine derivatives
containing an arylpiperazinyl function at the N3 side chain were
synthesized by using a Click reaction as the key step. Although
these compounds were prepared by combining the D1 and 5-
HT1A receptor agonistic pharmacophores, their binding potency
at the D1 receptor is generally lower, and most compounds
showed good to moderate affinity at both D2 and 5-HT1A
receptors. Compound 8h, containing 1-m-tolyl-benzazepine

4876
J. Zhang et al. / Bioorg. Med. Chem. 17 (2009) 4873–4880
Figure 2. [35S]GTP
c
S binding assays of compounds 8h and 13 at D1, D2, and 5-HT1A receptors.
Table 2
D1, D2, and 5-HT1A receptor binding and [35S]GTP
c
S studies of compounds 8h and 13a
Compound
D1
D2
5-HT1A
Cpd alone
Cpd + SKF38393
Cpd alone
Cpd + Quinpirole
Cpd alone
Emax (%)/EC50
Imax (%)/IC50
Emax (%)/EC50
Imax (%)/IC50
Emax (%)/EC50
8h
NA
100%/32.7
l
M
NA
100%/1.57
l
M
136%/200 nM
13
NA
70%/19.8
l
M
NA
100%/480 nM
125%/330 nM
SKF38393
Quinpirole
5-HT
100%/300 nM
—
—
—
—
—
—
—
—
—
—
—
100%/240 nM
—
100%/150 nM
a
Values are means of three to five experiments, and NA indicates no significant stimulation was detected; Dash lines indicate not applicable.
scaffold and 2-methoxyphenylpiperazine core, displayed good
affinity at all tested receptors, with Ki values of 144, 80, and
133 nM, for the D1, D2, and 5-HT1A receptors, respectively. Com-
pound 13 with the triazole moiety formed differently from that
in 8h showed the highest affinity at the D2 receptor with Ki value
of 19 nM. This compound also showed good affinity at the 5-HT1A
receptor (Ki, 105 nM), and moderate affinity at the D1 receptor (Ki,
551 nM). Functional assays indicated that both compounds 13 and
8h are antagonists at D1 and D2 receptors, whereas full agonistic
activity at the 5-HT1A receptor was observed. In agreement with
the binding affinity, compound 13 is a high efficacy D2 antagonist
and 5-HT1A agonist.
5. Experimental
5.1. Chemistry
Melting points were determined on a Thomas–Hoover capillary
tube apparatus and are reported uncorrected. 1H and 13C NMR
spectra were recorded on a Brucker AC300 spectrometer using tet-
ramethylsilane as an internal reference. Element analyses, per-
formed by the Analytic Lab, SIMM, were within 0.4% of
theoretical values. Analytical thin-layer chromatography (TLC)
was carried out on 0.2-mm Kieselgel 60F 254 silica gel plastic
sheets (EM Science, Newark). The column output was monitored
with TLC. Yields of all the reactions were not optimized. Com-
pounds 4 (20-tolyl or 30-tolyl) were prepared using a reported
procedure.27
Since it has been well documented19,20,22 that a chloro-sub-
stituent at the C7 of the benzazepine core is a determinant
for D1 receptor antagonistic activity, the antagonism of com-
pounds 8h and 13 at the D1 receptor suggested that an addi-
tional antagonistic binding site for the D1 receptor may exist
at the N3 side chain. Further, our current results also indicated
that a lipophilic binding site for the D2 receptor is existed at the
N3 side chain in the D1 receptor agonistic scaffold, 1-aryl-3-ben-
zazepine skeleton.
5.1.1. General procedure for the preparation of N3-substituted
benzazepines 5 and 11
Alkynol tosylate (1.1 equiv) was added to a solution of benzaze-
pine 4 (1.0 equiv) and K2CO3 (2.0 equiv) in acetonitrile. The

J. Zhang et al. / Bioorg. Med. Chem. 17 (2009) 4873–4880
4877
mixture was stirred at 90 °C overnight. After cooling to rt, the mix-
ture was filtered and concentrated. The crude material was puri-
fied by silica gel chromatography to give the corresponding
benzazepines 5 and 11.
5.1.2.3. 3-(4-(3-Azidopropyl)piperazin-1-yl)benzo[d]isothiazole
(6, Ar = 3-BIT, m = 1). Yield 73% (for two steps); 1H NMR
(300 MHz, CDCl3): d = 7.68 (dd, J = 0.9, 8.1 Hz, 1H), 7.62 (dd,
J = 1.2, 7.5 Hz, 1H), 7.55 (dt, J = 1.5, 7.5 Hz, 1H), 7.27 (m, 1H), 3.20
(t, J = 6.6 Hz, 2H), 3.09 (t, J = 4.8 Hz, 4H), 2.52 (t, J = 5.1 Hz, 4H),
2.41 (t, J = 6.9 Hz, 2H), 1.73 ppm (m, 2H).
5.1.1.1. 7,8-Dimethoxy-3-(pentyn-4-yl)-1-o-tolyl-2,3,4,5-tetra-
hydro-1H-benzo[d]azepine (5, R = 20-Me, n = 1). Yield 61%; 1H
NMR (300 MHz, CDCl3): d = 7.21 (m, 4H), 6.68 (s, 1H), 5.92 (s,
1H), 4.46 (d, J = 9.0 Hz, 1H), 3.85 (s, 3H), 3.47 (s, 3H), 3.29 (m,
2H), 3.14 (dd, J = 6.6, 12.0 Hz, 1H), 2.82 (m, 4H), 2.41 (m, 3H),
2.16 (s, 3H), 1.96 (t, J = 2.4 Hz, 1H), 1.73 ppm (m, 2H).
5.1.2.4. 3-(3-Azidopropyl)-7,8-dimethoxy-1-o-tolyl-2,3,4,5-tet-
rahydro-1H-benzo[d]azepine (10, R = 20-Me, n = 1). Yield 71%
(for two steps); 1H NMR (300 MHz, CDCl3): d = 7.22 (m, 4H), 6.68
(s, 1H), 5.93 (s, 1H), 4.46 (d, J = 9.0 Hz, 1H), 3.85 (s, 3H), 3.47 (s,
3H), 3.29 (m, 4H), 3.12 (dd, J = 6.3, 11.7 Hz, 1H), 2.77 (m, 2H),
2.62 (t, J = 6.6 Hz, 2H), 2.33 (t, J = 11.1 Hz, 1H), 2.17 (s, 3H),
1.78 ppm (m, 2H).
5.1.1.2. 7,8-Dimethoxy-3-(pent-4-ynyl)-1-m-tolyl-2,3,4,5-tetra-
hydro-1H-benzo [d]azepine (5, R = 30-Me, n = 1). Yield 75%; 1H
NMR (300 MHz, CDCl3): d = 7.23 (m, 1H), 7.06 (d, J = 7.5 Hz, 1H),
6.98 (m, 2H), 6.67 (s, 1H), 6.30 (s, 1H), 4.23 (dd, J = 3.0, 6.0 Hz,
1H), 3.89 (s, 3H), 3.72 (s,3H), 3.00 (m, 3H), 2.81 (m, 2H), 2.56 (m,
3H), 2.34 (s, 3H), 2.20 (m, 2H), 1.94 (t, J = 2.7 Hz, 1H), 1.72 ppm
(m, 2H).
5.1.3. General procedure for Click reaction28
A mixture of alkyne (1.0 equiv), CuI (0.2 equiv), and azide
(1.1 equiv) in THF (5 mL) was stirred for 5 min, and then N,N-diiso-
propylethylamine (DIPEA, 5.0 equiv) was added slowly, and the
mixture was stirred at rt overnight. The solution was filtered and
concentrated to give the crude material, which was chromato-
graphed to afford the desired cyclization products 7, 12, and 16.
5.1.1.3. 3-(But-3-ynyl)-7,8-dimethoxy-1-m-tolyl-2,3,4,5-tetrahy-
dro-1H-benzo[d]azepine (5, R = 30-Me, n = 0). Yield 74%; 1H NMR
(300 MHz, CDCl3): d = 7.24 (m, 1H), 7.03 (m, 3H), 6.68 (s, 1H), 6.30
(s, 1H), 4.24 (m, 1H), 3.87 (s, 3H), 3.64 (s, 3H), 3.11 (m, 2H), 2.90 (m,
5H), 2.62 (t, J = 10.2 Hz, 1H), 2.38 (m, 5H), 1.97 ppm (t, J = 2.4 Hz,
1H).
5.1.3.1. 7,8-Dimethoxy-3-(3-(1-(3-(4-(pyrimidin-2-yl)piperazin-
1-yl)propyl)-1H-1,2,3-triazol-4-yl)propyl)-1-o-tolyl-2,3,4,5-tet-
rahydro-1H-benzo[d]azepine (7, R = 20-Me, n = m = 1, Ar = 2-
Pym). Yield 85%; 1H NMR (300 MHz, CDCl3): d = 8.28 (d,
J = 4.8 Hz, 2H), 7.21 (m, 5H), 6.66 (s, 1H), 6.46 (t, J = 4.8 Hz, 1H),
5.89 (s, 1H), 4.45 (d, J = 9.0 Hz, 1H), 4.38 (t, J = 6.9 Hz, 2H), 3.82
(m, 7H), 3.45 (s, 3H), 3.28 (m, 2H), 3. 13 (dd, J = 6.3, 12.3 Hz, 1H),
2.74 (m, 4H), 2.59 (t, J = 7.5 Hz, 2H), 2.44 (t, J = 4.8 Hz, 4H), 2.33
(m, 3H), 2.14 (s, 3H), 2.06 (m, 2H), 1.88 ppm (m, 2H).
5.1.1.4. 3-(Hex-5-ynyl)-7,8-dimethoxy-1-m-tolyl-2,3,4,5-tetra-
hydro-1H-benzo[d]azepine (5, R = 30-Me, n = 2). Yield 79%; 1H
NMR (300 MHz, CDCl3): d = 7.22 (m, 1H), 7.06 (d, J = 7.8 Hz, 1H),
6.99 (m, 2H), 6.68 (s, 1H), 6.28 (s, 1H), 4.24 (dd, J = 2.4, 6.6 Hz,
1H), 3.86 (s, 3H), 3.63 (s, 3H), 3.01 (m, 3H), 2.83 (m, 2H), 2.50
(m, 3H), 2.34 (s, 3H), 2.19 (dt, J = 2.7, 6.9 Hz, 2H), 1.93 (t,
J = 2.7 Hz, 1H), 1.57 ppm (m, 4H).
5.1.3.2. 7,8-Dimethoxy-3-(3-(1-(3-(4-(2-methoxyphenyl)pipera-
zin-1-yl)propyl)-1H-1,2,3-triazol-4-yl)propyl)-1-o-tolyl-2,3,4,5-
tetrahydro-1H-benzo[d]azepine (7, R = 20-Me, n = m = 1, Ar = 2-
MeO-Ph). Yield 82%; 1H NMR (300 MHz, CDCl3): d = 7.21 (m, 4H),
6.93 (m, 4H), 6.67 (s, 1H), 5.90 (s, 1H), 4.46 (d, J = 9.3 Hz, 1H), 4.38
(t, J = 6.6 Hz, 2H), 3.85 (s, 3H), 3.84 (s, 3H), 3.45 (s, 3H), 3.29 (m,
2H), 3.13 (m, 5H), 2.75 (m, 4H), 2.60 (m, 6H), 2.34 (m, 3H), 2.15
(s, 3H), 2.07 (m, 2H), 1.90 ppm (m, 2H).
5.1.1.5. 1-(2-Methoxyphenyl)-4-(pent-4-ynyl)piperazine (11,
Ar = 2-MeO-Ph, m = 1). Yield 76%; 1H NMR (300 MHz, CDCl3):
d = 6.91 (m, 4H), 3.84 (s, 3H), 3.08 (br s, 4H), 2.64 (br s, 4H),
2.49 (t, J = 7.2 Hz, 2H), 2.24 (dt, J = 2.7, 7.2 Hz, 2H), 1.95 (t,
J = 2.7 Hz, 1H), 1.74 ppm (m, 2H).
5.1.2. General procedure for the preparation of azides 6 and 10
The corresponding arylpiperazines or benzazepine 4 (1.0 equiv),
chloroalkyl bromide (10.0 equiv), and K2CO3 (2.0 equiv) were stir-
red at 50 °C for 2 h. Then the solution was filtered and concen-
trated. The crude material was chromatographed to give the
corresponding chloride derivatives that were used directly for
the next step.
The mixture of the chlorides obtained above (1.0 equiv) and
NaN3 (2.0 equiv) was dissolved in DMSO (5 mL) and stirred at
100 °C for 2 h. After cooling to rt, the reaction mixture was diluted
with CHCl3, and washed with water, brine, and dried. The solution
was concentrated and chromatographed to give the corresponding
azides 6 and 10.
5.1.3.3. 7,8-Dimethoxy-3-(3-(1-(3-(4-(pyrimidin-2-yl)piperazin-
1-yl)propyl)-1H-1,2,3-triazol-4-yl)propyl)-1-m-tolyl-2,3,4,5-tet-
rahydro-1H-benzo[d]azepine (7, R = 30-Me, n = m = 1, Ar = 2-
Pym). Yield 88%; 1H NMR (300 MHz, CDCl3): d = 8.28 (d,
J = 4.8 Hz, 2H), 7.23 (m, 2H), 7.00 (m, 3H), 6.65 (s, 1H), 6.47 (t,
J = 7.8 Hz, 1H), 6.24 (s, 1H), 4.83 (t, J = 6.9 Hz, 2H), 4.23 (d,
J = 6.9 Hz, 1H), 3.82 (m, 7H), 3.60 (s, 3H), 2.86 (m, 6H), 2.45 (m,
10H), 2.31 (s, 3H), 2.07 (m, 2H), 1.88 ppm (m, 2H).
5.1.3.4. 7,8-Dimethoxy-3-(3-(1-(3-(4-(2-methoxyphenyl)pipera-
zin-1-yl)propyl)-1H-1,2,3-triazol-4-yl)propyl)-1-m-tolyl-2,3,4,5-
tetrahydro-1H-benzo[d]azepine (7, R = 30-Me, n = m = 1, Ar = 2-
MeO-Ph). Yield 99%; 1H NMR (300 MHz, CDCl3): d = 7.22 (m, 1H),
6.95 (m, 7H), 6.66 (s, 1H), 6.25 (s, 1H), 4.38 (t, J = 6.6 Hz, 2H), 4.25
(d, J = 7.5 Hz, 1H), 3.85 (s, 6H), 3.60 (s, 3H), 2.68 (m, 18H), 2.39 (t,
J = 6.6 Hz, 2H), 2.32 (s, 3H), 2.05 (m, 2H), 1.90 ppm (m, 2H).
5.1.2.1. 2-(4-(3-Azidopropyl)piperazin-1-yl)pyrimidine (6, Ar =
2-Pym, m = 1). Yield 75% (for two steps); 1H NMR (300 MHz,
CDCl3): d = 8.26 (d, J = 4.5 Hz, 2H), 6.43 (t, J = 4.8 Hz, 1H), 3.79 (t,
J = 4.8 Hz, 4H), 3.33 (t, J = 6.9 Hz, 2H), 2.43 (m, 6H), 1.76 ppm (m,
2H).
5.1.3.5.
3-(4-(3-(4-(3-(7,8-Dimethoxy-1-m-tolyl-4,5-dihydro-
1H-benzo[d]azepin-(2H)-yl)propyl)-1H-1,2,3-triazol-1-yl)pro-
pyl)piperazin-1-yl)benzo[d]isothiazole (7, R = 3’-Me, n = m = 1,
Ar = 3-BIT). Yield 84%; 1H NMR (300 MHz, CDCl3): d = 7.67 (d,
J = 7.5 Hz, 1H), 7.62 (d, J = 7.5 Hz, 1H), 7.55 (dt, J = 1.2, 7.5 Hz, 1H),
7.28 (m, 1H), 7.22 (m, 2H), 7.04 (d, J = 7.5 Hz, 1H), 6.97 (m, 2H), 6.66
(s, 1H), 6.24 (s, 1H), 4.33 (t, J = 6.9 Hz, 2H), 4.23 (d, J = 7.2 Hz, 1H),
5.1.2.2. 1-(3-Azidopropyl)-4-(2-methoxyphenyl)piperazine (6,
Ar = 2-MeO-Ph, m = 1). Yield 70% (for two steps); 1H NMR
(300 MHz, CDCl3): d = 6.95 (m, 3H), 6.86 (m, 1H), 3.86 (s, 3H),
3.36 (t, J = 6.9 Hz, 2H), 3.09 (br s, 4H), 2.65 (br s, 4H), 2.49 (t,
J = 6.9 Hz, 2H), 1.81 ppm (m, 2H).

4878
J. Zhang et al. / Bioorg. Med. Chem. 17 (2009) 4873–4880
3.85 (s, 3H), 3.60 (s, 3H), 3.03 (m, 7H), 2.83 (m, 2H), 2.69 (t, J = 7.2 Hz,
2H), 2.52 (m, 7H), 2.32 (m, 5H), 2.01 (m, 2H), 1.87 ppm (m, 2H).
5.1.4.2. 3-(3-(1-(3-(4-(2-Methoxyphenyl)piperazin-1-yl)propyl)-
1H-1,2,3-triazol-4-yl)propyl)-1-o-tolyl-2,3,4,5-tetrahydro-1H-
benzo[d]azepine-7,8-diol (8b). Yield 17%; 1H NMR (300 MHz,
CD3OD + CDCl3): d = 7.45 (s, 1H), 7.07 (m, 3H), 6.91 (m, 2H), 6.78
(m, 3H), 6.53 (s, 1H), 5.76 (s, 1H), 4.64 (d, J = 9.0 Hz, 1H), 4.27 (t,
J = 6.9 Hz, 2H), 3.74 (s, 3H), 3.44 (m, 3H), 3.10 (m, 7H), 2.61 (m,
8H), 2.35 (t, J = 6.9 Hz, 2H), 2.00 ppm (m, 7H); 13C NMR
(100 MHz, CD3OD + CDCl3): d = 152.0, 146.1, 142.7, 140.5, 136.2,
130.5, 126.9, 126.6, 126.1, 123.2, 121.7, 120.8, 118.0, 116.4,
114.4, 111.1, 60.4, 57.9, 55.0, 54.7, 54.6, 52.9, 50.0, 48.0, 41.3,
32.5, 26.7, 26.0, 24.2, 22.6, 19.1 ppm; MS (EI) m/z 610 [M+]; Anal.
Calcd for C36H46N6O3ꢁ1.5HBrꢁ0.5H2O: C, 58.34; H, 6.60; N, 11.34.
Found: C, 58.60; H, 6.56; N, 11.04.
5.1.3.6. 7,8-Dimethoxy-3-(2-(1-(3-(4-(2-methoxyphenyl)pipera-
zin-1-yl)propyl)-1H-1,2,3-triazol-4-yl)ethyl)-1-m-tolyl-2,3,4,5-
tetrahydro-1H-benzo[d]azepine (7, R = 30-Me, n = 0, m = 1,
Ar = 2-MeO-Ph). Yield 96%; 1H NMR (300 MHz, CDCl3): d = 7.20
(m, 1H), 7.13 (s, 1H), 6.96 (m, 6H), 6.85 (m, 1H), 6.67 (s, 1H),
6.33 (s, 1H), 4.31 (m, 3H), 3.86 (s, 3H), 3.84 (s, 3H), 3.64 (s, 3H),
3.14 (m, 6H), 2.89 (m, 7H), 2.63 (m, 5H), 2.38 (t, J = 7.2 Hz, 2H),
2.27 (s, 3H), 2.04 ppm (m, 2H).
5.1.3.7. 7,8-Dimethoxy-3-(4-(1-(3-(4-(2-methoxyphenyl)pipera-
zin-1-yl)propyl)-1H-1,2,3-triazol-4-yl)butyl)-1-m-tolyl-2,3,4,5-
tetrahydro-1H-benzo[d]azepine (7, R = 30-Me, n = 2, m
= 1,
5.1.4.3. 3-(3-(1-(3-(4-(2-Hydroxyphenyl)piperazin-1-yl)propyl)-
1H-1,2,3-triazol-4-yl)propyl)-1-o-tolyl-2,3,4,5-tetrahydro-1H-
benzo[d]azepine-7,8-diol (8c). Yield 32%; 1H NMR (300 MHz,
CD3OD): d = 7.86 (s, 1H), 7.18 (m, 4H), 6.92 (m, 2H), 6.82 (m, 2H),
6.68 (s, 1H), 5.86 (s, 1H), 4.71 (d, J = 9.3 Hz, 1H), 4.40 (t,
J = 6.6 Hz, 2H), 3.62 (m, 2H), 3.40 (m, 1H), 3.28 (m, 1H), 3.01 (br
s, 6H), 2.77 (m, 8H), 2.50 (t, J = 6.6 Hz, 2H), 2.12 ppm (m, 7H); 13C
NMR (100 MHz, CD3OD): d = 152.4, 147.9, 145.4, 145.3, 141.8,
140.8, 138.0, 134.9, 132.4, 131.8, 128.9, 128.8, 128.2, 126.2,
124.4, 121.6, 121.1, 118.5, 116.9, 116.3, 62.1, 59.7, 56.4, 56.3,
54.6, 51.9, 42.7, 33.3, 28.1, 25.6, 24.1, 20.1 ppm; MS (EI) m/z 596
[M+]; Anal. Calcd for C35H44N6O3ꢁ2.25HBr: C, 53.98; H, 5.99; N,
10.79. Found: C, 54.30; H, 6.00; N, 10.42.
Ar = 2-MeO-Ph)
.
Yield 91%; 1H NMR (300 MHz, CDCl3): d = 7.27
(s, 1H), 7.22 (m, 1H), 6.95 (m, 7H), 6.66 (s, 1H), 6.24 (s, 1H), 4.39
(t, J = 7.2 Hz, 2H), 4.25 (d, J = 7.5 Hz, 1H), 3.85 (s, 6H), 3.60 (s,
3H), 6.90 (m, 10H), 2.63 (brs, 4H), 2.47 (m, 5H), 2.32 (s, 3H), 2.11
(m, 3H), 1.62 ppm (m, 4H).
5.1.3.8. 7,8-Dimethoxy-3-(3-(4-(3-(4-(2-methoxyphenyl)pipera-
zin-1-yl)propyl)-1H-1,2,3-triazol-1-yl)propyl)-1-o-tolyl-2,3,4,5-
tetrahydro-1H-benzo[d]azepine (12, R = 20-Me, n = m = 1, Ar = 2-
MeO-Ph). Yield 97%; 1H NMR (300 MHz, CDCl3): d = 7.21 (m, 5H),
6.93 (m, 4H), 6.68 (s, 1H), 5.95 (s, 1H), 4.39 (m, 3H), 3.86 (s, 3H),
3.85 (s, 3H), 3.48 (s, 3H), 3.25 (m, 2H), 3.07 (m, 5H), 2.73 (m,
8H), 2.59 (m, 4H), 2.34 (t, J = 11.4 Hz, 1H), 2.18 (s, 3H), 2.07 (m,
2H), 1.90 ppm (m, 2H).
5.1.4.4.
3-(3-(1-(3-(4-(Pyrimidin-2-yl)piperazin-1-yl)propyl)-
1H-1,2,3-triazol-4-yl)propyl)-1-m-tolyl-2,3,4,5-tetrahydro-1H-
benzo[d]azepine-7,8-diol (8d). Yield 34%; 1H NMR (300 MHz,
CD3OD): d = 8.30 (d, J = 4.8 Hz, 2H), 7.77 (s, 1H), 7.23 (t, J = 7.5 Hz,
1H), 7.07 (d, J = 7.8 Hz, 1H), 6.97 (m, 2H), 6.59 (m, 2H), 6.09 (s,
1H), 4.42 (t, J = 6.9 Hz, 2H), 4.27 (d, J = 7.8 Hz, 1H), 3.77 (t,
J = 4.8 Hz, 4H), 3.15 (m, 4H), 2.67 (m, 6H), 2.45 (t, J = 5.1 Hz, 4H),
2.35 (m, 5H), 2.09 (m, 2H), 1.95 ppm (m, 2H); 13C NMR
(100 MHz, CD3OD): d = 163.3, 159.5, 148.4, 144.9, 144.8, 144.2,
140.1, 136.2, 132.6, 130.6, 130.3, 129.0, 127.0, 124.3, 118.6,
117.8, 111.8, 62.2, 59.7, 56.8, 56.6, 54.4, 48.9, 34.8, 28.6, 26.7,
24.4, 22.1 ppm; MS (EI) m/z 582 [M+]; Anal. Calcd for
C33H42N8O2ꢁ1.2HBrꢁ0.9H2O: C, 56.94; H, 6.52; N, 16.10. Found: C,
57.38; H, 6.51; N, 15.72.
5.1.3.9. N,N-Diethyl-3-(1-(3-(4-(2-methoxyphenyl)piperazin-1-
yl)propyl)-1H-1,2,3-triazol-4-yl)propan-1-amine
(16). Yield
90%; 1H NMR (300 MHz, CDCl3): d = 7.30 (s, 1H), 6.90 (m, 4H),
4.39 (t, J = 6.9 Hz, 2H), 3.83 (s, 3H), 3.11 (br s, 4H), 2.70 (t,
J = 7.5 Hz, 2H), 2.60 (br s, 4H), 2.50 (m, 6H), 2.38 (t, J = 6.9 Hz,
2H), 2.07 (m, 2H), 1.82 (m, 2H), 0.99 (t, J = 6.9 Hz, 6H); 13C NMR
(100 MHz, CDCl3) d 151.8, 147.4, 140.8, 122.6, 120.7, 120.6,
117.7, 110.8, 55.0, 54.4, 53.0, 51.8, 50.3, 47.6, 46.5, 27.1, 26.4,
23.3, 11.3 ppm; MS (EI) m/z 414 [M+]; HRMS: m/z [M+] calcd for
C23H38N6O: 414.3107, found: 414.3099.
5.1.4. General procedure for O-demethylation of compounds 7
and 12
A solution of 7 or 12 (1.0 equiv) in 9 mL dry CH2Cl2 was stirred
at ꢀ78 °C under nitrogen for 30 min, then 1 M BBr3 in CH2Cl2
(3.0 equiv) was added slowly. The mixture was stirred for addi-
tional 10 min at ꢀ78 °C and then at rt for 20 min. The reaction
was quenched with MeOH at ꢀ78 °C for 1 h, and concentrated.
The obtained residue was treated with MeOH again and concen-
trated. The crude material was purified with preparative TLC to
give the final compounds 8 and 13.
5.1.4.5. 3-(3-(1-(3-(4-(2-Methoxyphenyl)piperazin-1-yl)propyl)-
1H-1,2,3-triazol-4-yl)propyl)-1-m-tolyl-2,3,4,5-tetrahydro-1H-
benzo[d]azepine-7,8-diol (8e). Yield 29%; 1H NMR (300 MHz,
CDCl3 + CD3OD): d = 7.45 (s, 1H), 7.13 (t, J = 7.2 Hz, 1H), 6.88 (m,
7H), 6.55 (s, 1H), 6.00 (s, 1H), 4.38 (d, J = 8.4 Hz, 1H), 4.28 (t,
J = 7.8 Hz, 2H), 3.76 (s, 3H), 3.55 (m, 2H), 3.36 (m, 1H), 3.10 (m,
6H), 2.80 (m, 2H), 2.65 (m, 7H), 2.38 (t, J = 7.5 Hz, 2H), 2.21 (s,
3H), 2.00 ppm (m, 4H); 13C NMR (100 MHz, CDCl3 + CD3OD):
d = 152.0, 146.1, 142.6, 141.3, 140.4, 138.3, 134.1, 130.2, 128.8,
128.5, 127.5, 125.1, 123.2, 121.8, 120.8, 118.1, 116.7, 115.8,
111.0, 61.0, 59.8, 57.7, 55.1, 54.6, 52.9, 49.9, 48.0, 45.7, 32.2,
29.4, 26.6, 24.2, 22.5, 21.1 ppm; MS (EI) m/z 610 [M+]; Anal. Calcd
for C36H46N6O3ꢁ2.3HBrꢁ0.2H2O: C, 54.02; H, 6.13; N, 10.50. Found:
C, 54.36; H, 6.11; N, 10.07.
5.1.4.1.
3-(3-(1-(3-(4-(Pyrimidin-2-yl)piperazin-1-yl)propyl)-
1H-1,2,3-triazol-4-yl) propyl)-1-o-tolyl-2,3,4,5-tetrahydro-1H-
benzo[d]azepine-7,8-diol (8a). Yield 35%; 1H NMR (300 MHz,
CD3OD): d = 8.30 (d, J = 4.5 Hz, 2H), 7.82 (s, 1H), 7.20 (m, 4H),
6.66 (s, 1H), 6.58 (t, J = 4.8 Hz, 1H), 5.85 (s, 1H), 4.67 (dd, J = 3.6,
9.0 Hz, 1H), 4.43 (t, J = 6.6 Hz, 2H), 3.77 (m, 4H), 3.62 (m, 2 H),
3.34 (m, 1H), 3.24 (m, 1H), 3.04 (t, J = 7.8 Hz, 2H), 2.86 (dd,
J = 6.0, 15.6 Hz, 1H), 2.75 (m, 3H), 2.47 (t, J = 5.1 Hz, 4H), 2.38 (t,
J = 6.9 Hz, 2H), 2.14 (s, 3H), 2.03 ppm (m, 4H); 13C NMR
(100 MHz, CD3OD): d = 163.2, 159.6, 147.9, 145.4, 141.9, 137.9,
132.3, 128.9, 128.7, 128.2, 124.4, 118.5, 116.3, 111.8, 62.3, 59.8,
56.5, 54.4, 45.0, 43.0, 33.4, 28.5, 25.7, 24.1, 20.5, 20.1 ppm; MS
(EI) m/z 582 [M+]; Anal. Calcd for C33H42N8O2ꢁ1.2HBrꢁ1.0H2O: C,
56.80; H, 6.53; N, 16.06. Found: C, 57.25; H, 6.87; N, 15.45.
5.1.4.6. 3-(3-(1-(3-(4-(Benzo[d]isothiazol-3-yl)piperazin-1-yl)-
propyl)-1H-1,2,3-triazol-4-yl)propyl)-1-m-tolyl-2,3,4,5-tetrahy-
dro-1H-benzo[d]azepine-7,8-diol (8f). Yield 28%; 1H NMR
(300 MHz, CD3OD): d = 7.97 (d, J = 8.4 Hz, 1H), 7.88 (d, J = 8.4 Hz,
1H), 7.76 (s, 1H), 7.51 (t, J = 7.5 Hz, 1H), 7.40 (t, J = 7.5 Hz, 1H),
7.20 (t, J = 8.1 Hz, 1H), 7.04 (d, J = 7.8 Hz, 1H), 6.92 (m, 2H), 6.60
(s, 1H), 6.09 (s, 1H), 4.43 (t, J = 7.2 Hz, 2H), 4.26 (d, J = 8.4 Hz,
1H), 3.57 (m, 4H), 3.25 (m, 1H), 3.08 (m, 3H), 2.65 (m, 10H), 2.41

J. Zhang et al. / Bioorg. Med. Chem. 17 (2009) 4873–4880
4879
(t, J = 6.9 Hz, 2H), 2.29 (s, 3H), 2.10 (m, 2H), 1.94 ppm (m, 2H); 13
C
GF/B glass fiber filter and subsequent washing with cold buffer
(50 mM Tris, 5 mM EDTA, pH 7.4) using a Brandel 24-well cell har-
vester. Scintillation cocktail was added and the radioactivity was
determined in a MicroBeta liquid scintillation counter. The IC50
and Ki values were calculated by nonlinear regression (PRISM,
Graphpad, San Diego, CA) using a sigmoidal function.
NMR (100 MHz, CD3OD): d = 165.8, 154.3, 148.7, 144.7, 139.9,
136.7, 133.1, 130.7, 130.2, 129.6, 129.5, 128.8, 127.0, 125.9,
125.7, 124.2, 122.2, 118.5, 117.8, 62.7, 62.5, 59.8, 56.9, 56.6, 54.4,
51.4, 49.1, 35.3, 28.6, 27.0, 24.5, 22.1 ppm; MS (EI) m/z 637 [M+];
Anal. Calcd for C36H43N7O2Sꢁ1.3HBrꢁ1.0H2O: C, 56.67; H, 6.38; N,
12.85. Found: C, 56.77; H, 6.09; N, 12.60.
5.3. [35S]GTP
cS binding
5.1.4.7. 3-(2-(1-(3-(4-(2-Methoxyphenyl)piperazin-1-yl)propyl)-
1H-1,2,3-triazol-4-yl)ethyl)-1-m-tolyl-2,3,4,5-tetrahydro-1H-
benzo[d]azepine-7,8-diol (8g). Yield 42%; 1H NMR (300 MHz,
CDCl3): d = 7.13 (t, J = 7.5 Hz, 1H), 7.00 (m, 3H), 6.86 (m, 5H), 6.70
(s, 1H), 6.13 (s, 1H), 4.21 (m, 3H), 3.84 (s, 3H), 2.97 (m, 12H),
2.64 (m, 6H), 2.39 (m, 2H), 2.25 (s, 3H), 2.00 ppm (m, 2H); 13C
NMR (100 MHz, CDCl3): d = 152.1, 145.5, 143.1, 142.9, 142.4,
140.6, 138.1, 134.9, 131.5, 129.1, 128.4, 127.1, 125.3, 123.2,
122.2, 120.9, 118.2, 116.8, 116.5, 111.2, 59.7, 57.3, 55.3, 54.7,
54.2, 53.1, 49.8, 48.2, 48.1, 33.6, 26.8, 22.9, 21.4 ppm; MS (EI) m/z
597 [M+H]+; Anal. Calcd for C35H44N6O3ꢁ0.65HBr: C, 64.74; H,
6.93; N, 12.94. Found: C, 64.94; H, 7.06; N, 12.58.
For detection of the agonism action of the compounds, the
35S]GTP
S binding assay was performed at 30 °C for 30 min con-
taining 10 g membrane protein in a final volume of 100 L with
various concentrations of the drug. The antagonism effects of the
compounds were tested in the presence of 10 M SKF38393 for
D1 receptor or 10 M quinpirole for D2 receptor. The binding buffer
contains 50 mM Tris, pH 7.5, 5 mM MgCl2, 1 mM EDTA, 100 mM
NaCl, 1 mM DTT, and 40 M GDP. The reaction was initiated by
adding [35S]GTP
S (final concentration of 0.1 nM). Nonspecific
binding was measured in the presence of 100 M Gpp(NH)p. The
reaction was terminated by the addition of 1 mL of ice-cold wash-
ing buffer (50 mM Tris, pH 7.5, 5 mM MgCl2, 1 mM EDTA, 100 mM
NaCl) and was rapidly filtered with GF/C glass fiber filters (What-
man) and washed three times. Radioactivity was determined by li-
quid scintillation counting.
[
c
l
l
l
l
l
c
l
5.1.4.8. 3-(4-(1-(3-(4-(2-Methoxyphenyl)piperazin-1-yl)propyl)-
1H-1,2,3-triazol-4-yl)butyl)-1-m-tolyl-2,3,4,5-tetrahydro-1H-
benzo[d]azepine-7,8-diol (8h). Yield 67%; 1H NMR (300 MHz,
CDCl3): d = 7.32 (s, 1H), 7.12 (m, 1H), 6.98 (m, 2H), 6.84 (m, 5H),
6.66 (s, 1H), 6.00 (s, 1H), 4.42 (m, 1H), 4.30 (m, 2H), 3.82 (s, 3H),
3.17 (m, 7H), 2.62 (m, 10H), 2.38 (m, 2H), 2.23 (s, 3H), 2.04 (m,
2H), 1.61 ppm (m, 4H); 13C NMR (100 MHz, CDCl3): d 152.1,
147.0, 143.0, 142.8, 141.9, 140.8, 138.3, 134.5, 130.5, 129.1,
128.6, 127.6, 125.3, 123.0, 121.4, 120.9, 118.1, 117.1, 116.1,
111.1, 60.2, 58.1, 55.3, 54.7, 53.1, 50.0, 48.0, 45.7, 32.4, 29.6,
26.9, 26.6, 24.7, 24.2, 21.4 ppm; MS (EI) m/z 624 [M+]; Anal. Calcd
for C37H48N6O3ꢁ0.45HBrꢁ2.0H2O: C, 63.73; H, 7.58; N, 12.05. Found:
C, 64.11; H, 7.39; N, 11.36.
Acknowledgments
This work was financially supported by grants from Chinese Na-
tional Science Foundation (30772625, 30672517), Shanghai Com-
mission of Science and Technology (07pj14104), and grant from
Ministry of Science and Technology (2007AA022163). Support
from Chinese Academy of Sciences, and Shanghai Institute of Mate-
ria Medica is also appreciated.
5.1.4.9. 3-(3-(4-(3-(4-(2-Methoxyphenyl)piperazin-1-yl)propyl)-
1H-1,2,3-triazol-1-yl)propyl)-1-o-tolyl-2,3,4,5-tetrahydro-1H-
benzo[d]azepine-7,8-diol (13). Yield 36%; 1H NMR (300 MHz,
CD3OD): d = 8.00 (s, 1H), 7.22 (m, 4H), 7.05 (m, 1H), 6.94 (m, 3H),
6.70 (s, 1H), 5.86 (s, 1H), 4.84 (d, J = 9.9 Hz, 1H), 4.51 (t,
J = 6.9 Hz, 2H), 3.84 (s, 3H), 3.74 (m, 2H), 3.47 (m, 3H), 3.34 (m,
3H), 3.21 (m, 8H), 2.92 (m, 2H), 2.81 (t, J = 7.5 Hz, 2H), 2.44 (m,
2H), 2.16 ppm (m, 5H); 13C NMR (100 MHz, CD3OD): d = 154.3,
147.9, 145.3, 145.2, 141.6, 141.2, 137.9, 134.8, 132.3, 131.7,
129.0, 128.8, 128.2, 125.8, 124.7, 122.7, 120.3, 118.5, 116.3,
113.4, 62.2, 57.9, 57.3, 56.6, 56.3, 54.1, 49.7, 49.0, 42.6, 33.1,
26.7, 25.4, 23.9, 20.2 ppm; MS (EI) m/z 610 [M+]; Anal. Calcd for
C36H46N6O3ꢁ1.2TFAꢁ1.0H2O: C, 60.24; H, 6.48; N, 10.98. Found: C,
60.49; H, 6.69; N, 11.02.
Supplementary data
Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.bmc.2009.06.019.
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