Carboline- and phenothiazine-derivated heterocycles as potent SIGMA-1 protein ligands

Article abstract of DOI:10.1016/j.ejmech.2014.10.053

Sigma 1 receptors are associated with neurodegenerative and psychiatric disorders. These receptors, via their chaperoning functions that counteract endoplasmic reticulum stress and block neurodegeneration, may serve as a target for a new generation of antidepressants or neuroprotective agents. The involvement of these receptors has also been observed in neuropathic pain and cancer. Only a few ligands, such as Igmesine and Anavex 2-73, have been involved in clinical trials. Thus the development of sigma 1 ligands is of interest to a new generation of drugs. Previous work in our lab underlined the potency of benzannulated bicyclic compounds as interesting ligands. Herein the work was extended to a series of novel tricyclic compounds. Carboline- and phenothiazine-derivated compounds were designed and synthesized. In vitro competition binding assays for sigma 1 and 2 receptors showed that most of them have high affinity for sigma 1 receptor (K_i = 2.5-18 nM), and selectivity toward sigma 2 receptor, without cytotoxic effects on SY5Y cells.

Full text of DOI:10.1016/j.ejmech.2014.10.053

European Journal of Medicinal Chemistry 89 (2015) 198e206
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Carboline- and phenothiazine-derivated heterocycles as potent
SIGMA-1 protein ligands
a
,
b
,
, c
Marion Donnier-Marechal , Paul-Emmanuel Larchanche b, Delphine Le Broc a
,
ꢀ
,
ꢀ a
Christophe Furman a c, Pascal Carato a *, Patricia Melnyk a
,
b,
, b, *
a Univ Lille Nord de France, F-59000 Lille, France
b UDSL, EA 4481, UFR Pharmacie, F-59000 Lille, France
c UDSL, EA 4483, UFR Pharmacie, F-59000 Lille, France
a r t i c l e i n f o
a b s t r a c t
Article history:
Sigma 1 receptors are associated with neurodegenerative and psychiatric disorders. These receptors, via
their chaperoning functions that counteract endoplasmic reticulum stress and block neurodegeneration,
may serve as a target for a new generation of antidepressants or neuroprotective agents. The involve-
ment of these receptors has also been observed in neuropathic pain and cancer. Only a few ligands, such
as Igmesine and Anavex 2-73, have been involved in clinical trials. Thus the development of sigma 1
ligands is of interest to a new generation of drugs. Previous work in our lab underlined the potency of
benzannulated bicyclic compounds as interesting ligands. Herein the work was extended to a series of
novel tricyclic compounds. Carboline- and phenothiazine-derivated compounds were designed and
synthesized. In vitro competition binding assays for sigma 1 and 2 receptors showed that most of them
have high affinity for sigma 1 receptor (Ki ¼ 2.5e18 nM), and selectivity toward sigma 2 receptor, without
cytotoxic effects on SY5Y cells.
Received 26 May 2014
Received in revised form
16 October 2014
Accepted 17 October 2014
Available online 18 October 2014
Keywords:
Sigma receptors
Carboline
Tetrahydrobenzofuropyridine
Phenoxazine
Phenothiazine
© 2014 Published by Elsevier Masson SAS.
1. Introduction
intra-mitochondrial calcium homoeostasis, ER stress and signal
transduction. Indeed, when ER Ca2þ is depleted, S1R chaperone IP3
Sigma 1 receptors (S1R) are unique non-opioid receptors located
at the endoplasmic reticulum (ER)emitochondrion interface
known as the MAM (mitochondrion associated endoplasmic re-
ticulum membrane) [1,2]. They are ubiquitously expressed in the
central nervous system, but are also found in peripheral organs
including lung, liver, kidney, eye and heart [1]. One S1R subtype
was cloned in 1996 [3]. It is an integral membrane protein of 223
amino acids with three hydrophobic domains [1]. In contrast, sigma
2 receptors (S2R) have not been cloned yet.
receptors localize at the MAM to ensure Ca2þ transmission between
the ER and mitochondria [4]. S1R are chaperones that correct
misfolded proteins, stabilize other ER proteins and regulate ER-
associated degradation (ERAD) [5]. Moreover, S1R localization is
dynamic, and S1R are able to translocate from the MAM to the
plasma membrane, where they may regulate a variety of functional
proteins, including ion channels, receptors and kinases. Thus these
receptors are involved in the regulation of numerous neurotrans-
mitter systems [4].
S1R, as an ER chaperone protein, is regulated by its association
with another chaperone protein, BiP (or GRP 78). S1R regulates
The involvement of S1R in depression [6,7], drug abuse [8],
neurodegenerative diseases [9,10], but also in pain [11,12] and
cancer proliferation [13], has been reviewed.
Several compounds have undergone clinical trials, but no se-
lective S1R ligands have so far been marketed, although many
drugs on the market show affinity for S1R. The selective S1R ligand
Igmesine has been studied in phase 3 clinical trials for depression
Abbreviations: ER, endoplasmic reticulum; MAM, mitochondrion associated
endoplasmic reticulum membrane; S1R, sigma 1 receptors; S2R, sigma 2 receptors;
BiP, immunoglobulin heavy-chain binding protein; GRP78, glucose-regulated pro-
tein 78.
* Corresponding authors. EA4481, UFR Pharmacie, 3 Rue du Pr Laguesse, BP83,
59006 Lille, France.
[14].
A phase 2 study is underway to evaluate the mixed
muscarinic-S1R agonist Anavex 2-73 in patients with Alzheimer's
disease [15,16].
E-mail addresses: marion.donnier-marechal@univ-lille2.fr (M. Donnier-
ꢀ
ꢀ
In previous work in our laboratory,
tetrahydroisoquinoline-hydantoin (tic-hydantoin)ederived
a
series of
Marechal), paul-emmanuel.larchanche@univ-lille2.fr (P.-E. Larchanche), delphine.
lebroc@univ-lille2.fr (D. Le Broc), christophe.furman@univ-lille2.fr (C. Furman),
pascal.carato@univ-lille2.fr (P. Carato), patricia.melnyk@univ-lille2.fr (P. Melnyk).
http://dx.doi.org/10.1016/j.ejmech.2014.10.053
0223-5234/© 2014 Published by Elsevier Masson SAS.

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M. Donnier-Marechal et al. / European Journal of Medicinal Chemistry 89 (2015) 198e206
199
compounds were described, with potent affinity for S1R in the
nanomolar range, high selectivity toward S2R and low cytotoxicity
(IC50 > 100 mM), underlining the interest of compounds 1 and 2
(Fig. 1) [17e20]. These ligands were evaluated in different phar-
macological models, and compound 1 showed high efficiency in a
cocaine addiction model [21], ischaemia and experimental auto-
immune encephalomyelitis [22].
tricyclic derivative [30]. The tricyclic cyclopentabenzoxazolone
derivatives (geh) were prepared in three steps starting from
benzoxazolone in a FriedeleCrafts reaction with a mixture of
aluminium trichloride in DMF and acetic acid chloride. Crotoni-
zation of the resulting derivatives with potassium hydroxide and
the corresponding benzaldehyde in ethanol, followed by cycli-
zation in polyphosphoric acid, yielded the tricyclic benzox-
azolone derivatives (geh) [31].
Tic-hydantoin derivatives
1 showed all of the properties
compatible with development except metabolic stability. The ma-
jority of metabolites resulted from tic-hydantoin instability. How-
ever, demethylated and debenzylated compounds have been
identified among all the metabolites of compound 1 (unpublished
data). An isoindoline moiety was therefore introduced to avoid this
metabolic instability (compound 2).
According to the general procedure shown in Scheme 2, a library
of 22 novel tricyclic derivatives was synthesized.
Starting from 3-bromo-1-chloropropane, reaction with appro-
priate amines yielded amino side chains 4aec and 5. These in-
termediates were prepared in CH3CN or DMF in the presence of
K2CO3 with the appropriate substituted amine such as N-methyl-
benzylamine, N-methyl-N-phenylethylamine, pyrrolidine or iso-
indoline (yields: 30e95%). The preparation of compounds 6e9
involved direct nucleophilic substitution using eight tricyclic scaf-
folds (aeh) under various conditions (procedure A, B, C) with
different bases (K2CO3, NaOH, NaH), in CH3CN or DMF. Tricyclic
moieties were all introduced on disubstituted amine (6aef, 7aef,
9aef), except for pyrrolidine (8aeb) and tricyclic benzoxazolone
(6geh), to give final compounds.
Procedure A (K2CO3, CH3CN) was adopted with tricyclic com-
pounds aed, geh and gave acceptable yields. This procedure A was
also applied to tricyclic scaffold c to afford compound 6c with very
low yield (3%), indeed we observed dramatic degradation in this
reaction. For the synthesis of compounds 7c and 9c, procedure B
was applied (NaOH, H2O, CH3CN), which gave better yields
(respectively 15% and 26%) with fewer degradation. For tricyclic
compounds e and f, a stronger base was necessary, procedure C
(NaH, DMF) was thus applied.
To solve this problem of instability, we looked for Tic-hydantoin
substitutes. Considering the hydrophobic moiety of the S1R phar-
macophore [23], the interest of heterocycles such as benzomor-
phanes, benzofurans [24], benzothiazolinones [25] and
benzoxazolinones [26] has been underlined. We recently described
benzannulated derivatives as potent and selective S1R ligands [27].
These compounds included benzoxazolinone, benzoxazinone,
benzothiazinone, benzimidazolone and benzimidazole as hetero-
cycles with propylbenzylmethylamine and propylisoindoline.
In this paper, we report our efforts to replace the tic-hydantoin
core with different tricyclic heterocycles. We decided to focus our
attention on the evaluation of fused cyclopentabenzoxazole as
tricyclic analogues of our previously described bicyclic ligands,
carboline and phenothiazine moieties. In the light of our previous
studies, we selected 3-(N-benzyl-N-methylamino)propyle and 3-
(isoindolin-2-yl)propyle moieties, already introduced on de-
rivatives 1, 2 [18,20]. 3-(N-phenylethyl-N-methylamino)propyle
and 3-(pyrrolidin-1-yl)propyle were also selected for this study
[27].
The structures of all final compounds were determined by 1H
NMR, 13C NMR and LC-MS analyses. Purity was evaluated under two
different HPLC conditions, and exceeded 96%. Results are presented
in the experimental section (SI). For the pharmacological evalua-
tion, all final products were converted into their water soluble
hydrochloride salts.
2. Chemistry
A series of tricyclic derivatives was synthesized by nucleo-
philic substitution of amino heterocycles with chloroalkylamino
side chains (Scheme 1). The heterocycles (aeh) were either
commercially available, such as 2,3,4,9-tetrahydro-1H-pyrido[3,4-
b] indole (beta carboline, a), phenoxazine (e) and phenothiazine
(f), or synthesized (Scheme 1). 1-Phenyl-2,3,4,9-tetrahydro-1H-
pyrido[3,4-b] indole (b) was synthesized, starting from trypt-
amine, benzaldehyde and trifluoroacetic acid in dichloromethane,
by PicteteSpengler reaction [28]. 2,3,4,5-Tetrahydro-1H-pyrido
[4,3-b]indole (gamma-carboline, c) was prepared in one step,
starting from phenyl hydrazine and 4-piperidinone in ethanol
[29]. 1,2,3,4-Tetrahydrobenzofuro[3,2-c]pyridine (d) was synthe-
sized in two steps from the corresponding O-phenylhydroxyl-
amine and ethyl-4-oxopiperidine-1-carboxylate in ethanol,
followed by deprotection of the carbamate to generate the
3. Results and discussion
3.1. Biological evaluations and SAR
S1R and S2R affinities were investigated in competition exper-
iments with radioligands according to the methods described by
Ganapathy et al. [32]. In the S1R assay, the selective ligand [3H]
(þ)-pentazocine was employed as the radioligand. Since a selective
S2R radioligand was not commercially available, the non-selective
ligand [3H]-DTG was employed in the S2R assay, in the presence
of an excess of non-tritiated (þ)-pentazocine, which selectively
occupies S1R. In both assays, Jurkat cell membranes were used as a
source of receptors.
O
O
N
N
N
N
N
N
O
O
1 (R):
1 (S):
2
Ki (nM)
S1R
S2R S2R/S1R
1 (R)
1 (S)
2
1000
496
141
110
7.1
4.5
545
5.3
103
Fig. 1. Structure of Tic-hydantoin-derivated compounds 1 and 2.

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M. Donnier-Marechal et al. / European Journal of Medicinal Chemistry 89 (2015) 198e206
200
NH2
NH
i
N
N
H
H
O
b
O
H
H
N
N
N
ii
iii
iv
NH2
NH2
R
N
O
O
N
O
H
H
c
d
H
H
N
H
N
H
N
R
N
vii
vi
v
O
O
O
O
O
O
O
O
O
O
g : R = H
h : R = Cl
O
Reagents and conditions: i: benzaldehyde, trifluoroacetic acid, CH2Cl2, rt, 16h (70%); ii: 4-
piperidonone, absolute ethanol, rt, 2h (64%); iii: ethyl-4-oxopiperidin-1-yl carboxylate, HCl, absolute
ethanol, reflux, 2h (84%); iv: potassium hydroxide, water, ethanol, reflux, 24h (94%); v: CH3COCl,
AlCl3, DMF, 80°C, 4h (75%); vi: potassium hydroxide, water, ethanol, rt, 16h (R= H, Cl: 85%); vii:
polyphosphoric acid, 120°C, 1h (R= H: 70% and R= Cl: 30%).
Scheme 1. Preparation of tricyclic compounds b, d, g, h.
The Ki values for S1R were determined from the corresponding
cyclopentabenzoxazolone. From the results obtained, it appears
that the replacement of the tic-hydantoin core with different
tricyclic derivatives can strongly affect the affinity for S1R and
S2R. Indeed, although a variety of modulations leads to a dra-
matic loss of S2R/S1R selectivity compared to the lead com-
pounds 1 and 2, high affinity for S1R is often maintained and
sometimes improved.
IC50 values for each compound 6e9. For compounds showing high
affinity for S1R, the Ki values for S2R and the S2R/S1R selectivity
ratios were also calculated (Table 1).
The tic-hydantoin moiety was replaced with various tricyclic
scaffolds such as beta- or gamma-carboline, tetrahy-
drobenzofuropyridine,
phenoxazine,
phenothiazine
and
Scheme 2. Synthesis of final compounds 6aeh, 7aef, 8aeb, 9aef.

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M. Donnier-Marechal et al. / European Journal of Medicinal Chemistry 89 (2015) 198e206
201
Table 1
obtained in our previously described Tic-hydantoin series (S2R/
S1R ¼ 141 for compound 1(R)) [17e19], or even in spiro[2]benzo-
pyran-1,40-piperidine series (S2R/S1R superior to 1000) [33].
Using phenoxazine as the heterocyclic moiety gave disap-
pointing results, with low affinity for S1R (Ki > 200 nM), and
without selectivity for compounds 6e and 9e. Derivative 7e showed
good affinity for S1R (Ki ¼ 18 nM), and moderate affinity for S2R
(Ki ¼ 80 nM); however it showed very low S2R/S1R selectivity (4).
Replacement of the oxygen atom by a sulphur atom resulted in
compounds 6f, 7f and 9f. Compared to the phenoxazine series,
phenothiazine compounds 6f, 7f and 9f showed better affinity for
S1R (Ki ¼ 11e28 nM) and S2R/S1R selectivity (8e28). Compounds
6geh, with cyclopentabenzoxazole moieties, displayed disap-
pointing affinity for S1R (Ki ¼ 72e160 nM) and no S2R/S1R selec-
tivity (0.3e2.5). We observed that introduction of a chlorine atom
on compound 6h, compared to 6g, induced more affinity for S2R
(Ki ¼ 50 nM) than for S1R (Ki ¼ 160 nM). Consequently, the other
disubstituted amines (7e9) were not introduced on tricyclic ben-
zoxazolone moieties g and h. The addition of a cyclopentyle ring to
afford the cyclopentabenzoxazole scaffold induced a dramatic loss
of affinity for S1R and selectivity for S2R compared to our previ-
ously reported benzoxazolinone compounds [27].
Affinity for S1R and S2R, and cytotoxicity evaluation of target compounds 6e9.
Compound
Ki (nM)
Ki (s2)/Ki (s1
)
IC50
(
m
M) SY5Y IC50/Ki (s1)
s1
s2
175
Haloperidol
11.9
4.5
7.1
5.3
6.2
4.3
18
2.0
>200
28
15
110
108
78
2.4
23
7
<5
<1
10
2.5
0.3
13
8
e
e
1(S)
1(R)
2
6a
6b
6c
6d
6e
6f
6g
6h
7a
7b
7c
7d
7e
7f
8a
8b
9a
9b
9c
9d
9e
9f
496a
764a
416a
15
>100
19.5
nd
19.7
9.3
16.9
68.9
71.9
17.1
>100
5.0
11.6
11.8
33.2
25.4
46.9
8.6
32.9
47.8
24.9
18.3
50.0
78.9
11.0
83.3
>20,000
2746
nd
3177
2163
939
34,450
<350
611
>1389
31
1611
1073
3018
10,160
2606
782
<160
1328
3112
2179
694
100
130
<10
180
280
180
50
91
93
200
78
80
310
>400
310
110
110
180
9.9
140
94
72
160
7.2
11
11
2.5
18
11
>200
36
8.0
8.4
72
18
31
4
28
e
9
14
13
2.5
0.9
<1
8
Among the N-methylbenzylamine, N-methyl-N-phenylethyl-
amine and isoindoline moieties, N-methyl-N-phenylethylamine
gave the best S2R/S1R selectivity (31 for 7d and 28 for 7f) in the
tricyclic series (7aef), with good affinity for S1R (Ki ¼ 2.5e18 nM)
and moderate affinity for S2R (Ki ¼ 78e310 nM).
11
>200
12
7173
<55
6942
Mean IC50 values for 2e3 independent experiments are shown with less than 10%
deviation.
a
Rat cerebral cortex membranes were used as a source of s2 receptors. nd: Not
3.2. Evaluation of cytotoxic effects
determined.
To determine the potential cytotoxic effects of our synthetic
derivatives, the human neuroblastoma cell line SY5Y was treated
with the whole compounds at different concentrations up to
The nature of the tricyclic moieties influences affinity for S1R
and S2R, and therefore the S2R/S1R selectivity. In the beta-
carboline series, with or without a phenyl ring at the 1 position,
compounds 6aeb, 7aeb, and 9aeb displayed good affinity for S1R
(Ki ¼ 4.3e11 nM) and moderate affinity for S2R (Ki ¼ 91e110 nM),
except for derivative 6a (Ki ¼ 15 nM). The S2R/S1R selectivity of
these compounds (from 2.4 to 23) was lower than the tic-hydantoin
derivatives (greater than 103). For beta-carboline derivatives with
pyrrolidine, the affinity for S1R was low for 8a (Ki > 200 nM) and
moderate for 8b (Ki ¼ 36 nM). Despite a moderate affinity, this last
compound displayed S2R/S1R selectivity (9) due to a low affinity for
S2R (Ki ¼ 310 nM). Therefore, the pyrrolidine side chain was not
introduced on the other heterocycles. Substitution with gamma-
carboline as heterocycle gave compounds 6c, 7c, and 9c. While
derivatives 6c and 7c showed good affinity for S1R, with Ki values of
18 and 11 nM respectively, compound 9c showed only moderate
affinity (Ki ¼ 72 nM). Compared to the beta-carboline series,
modulation of the position of the nitrogen atom with the gamma-
carboline derivatives induced lower affinity for S1R and S2R of
compounds 6c, 7c, and 9c (Ki ¼ 130e200 nM), but similar S2R/S1R
selectivity (values from 2.5 to 18). Replacement of the NH group in
the gamma-carboline by an oxygen atom resulted in the tetrahy-
drobenzofuropyridine series. This series showed high affinity for
S1R, with Ki values between 2.0 and 11 nM for compounds 6d, 7d
and 9d. Compared to the gamma carboline series, replacement of
the NH group by an oxygen atom in the tetrahydrobenzofuropyr-
idine compounds induced a significant increase in affinity for S1R,
but mainly for S2R, with a Ki value lower than 10 nM and a Ki value
of 9.9 for 6d and 9d respectively, leading to a loss of S2R/S1R
selectivity (<5 and 0.9). Therefore, among the 22 evaluated tricyclic
compounds, it is worth emphasizing that the best selectivity (S2R/
S1R ¼ 31) was obtained for compound 7d (Ki (S1R) ¼ 2.5 nM and Ki
(S2R) ¼ 78 nM). It has to be noted that better selectivity was
100
mM. Cell viability was calculated using a colorimetric MTT
assay. All tricyclic derivatives exhibited moderate to low cytotox-
icity (Table 1). Compound 6h showed the highest cytotoxicity,
5.0
m
M, with a low selectivity index, IC50(SY5Y)/Ki (S1R) ratio of 31.
M, but a very good
Compound 6b showed high cytotoxicity, 9.3
m
selectivity index, IC50(SY5Y)/Ki (S1R) ratio of 2163. Excellent selec-
tivity indexes were obtained with compounds 6d, 7d and 9d, with
ratios of 7173 to 34,450. All these derivatives contained tetrahy-
drobenzofuropyridine as the tricyclic moiety. The gamma-
carboline series, with compounds 6c and 9c, showed low selec-
tivity index, 939 and 694 respectively. Derivatives 6e, 6h, 8a and
9e showed both low affinities for S1R (superior to 160 nM) and low
selectivity index (IC50(SY5Y)/Ki
ratio: 31e350). Finally, com-
(S1R)
pounds (6aeb, 7aeb, 8aeb and 9aeb), with a beta-carboline
moiety, have intermediate but high selectivity index, with values
from 1073 to 3177.
4. Conclusion
S1R is an ER chaperone protein that has been implicated in
many neurodegenerative and neuropsychiatric disorders, and in
the response to cellular oxidative stress. Thus, its ligands may be of
interest as therapeutic agents. We synthesized compounds with
tricyclic scaffolds and various disubstituted amines, and evaluated
their affinity for sigma receptors and their cytotoxic effect on
neuronal cells. This study completed our previously work on ben-
zannulated bicyclic compounds as interesting S1R ligands. Most of
these tricyclic compounds showed high affinity for S1R and very
low cytotoxicity. Their selectivity indexes against S2R, ranging from
1 to 31, provide non-selective and selective sigma-1 ligands
respectively. This study identified compound 7d as a very efficient
and non-cytotoxic derivative. Though it has moderate selectivity

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M. Donnier-Marechal et al. / European Journal of Medicinal Chemistry 89 (2015) 198e206
202
for S2R, this compound 7d was selected for further in vivo evalua-
tions. Work is in progress.
J ¼ 7.1 Hz, 2H, CH2). LCMS (ESIþ): Calc. for [MþH]: 198.09; 200.09.
Found: 198.01; 199.96. HPLC (C4, 35 min): tR 8.01 min, PHPLC 96%;
HPLC (C18, 35 min): tR 14.69 min, PHPLC 96%.
5. Experimental section
5.1.2.2. 3-Chloro-N-methyl-N-phenylethylpropan-1-amine 4b [27].
Purification by column chromatography (DCM:MeOH(NH3), 97:3
(v/v)) was performed and enabled to collect the product as a
yellowish liquid (2.28 g, 65%). TLC: Rf 0.7 (DCM:MeOH(NH3), 90:10,
5.1. Chemistry
5.1.1. General
Chemicals and solvents were obtained from commercial sour-
ces, and used without further purification unless otherwise noted.
Reactions were monitored by TLC performed on MachereyeNagel
Alugram® Sil 60/UV254 sheets (thickness 0.2 mm). Purification of
products was carried out by either column chromatography or thick
layer chromatography. Column chromatography was carried out on
using MachereyeNagel silica gel (230e400 mesh). Thick layer
chromatography was performed on glass plates coated with
MachereyeNagel Sil/UV254 (thickness 2 mm), from which the pure
compounds were extracted with the following solvent system:
DCM/MeOH(NH3), 90:10. NMR spectra were recorded on a Bruker
DRX 300 spectrometer (operating at 300 MHz for 1Hand 75 MHz for
13C). Chemical shifts are expressed in ppm relative to either tetra-
methylsilane (TMS) or to residual proton signal in deuterated sol-
v/v). 1H NMR (300 MHz, CDCl3)
d: 7.33e7.27 (m, 2H, Haro); 7.42e7.16
(m, 3H, Haro); 3.57 (t, J ¼ 7.2 Hz, 2H, CH2); 2.78 (m, 2H, CH2); 2.63
(m, 2H, CH2); 2.56 (t, J ¼ 7.1 Hz, 2H, CH2); 2.31 (s, 3H, CH3); 1.93 (p,
J ¼ 7.0 Hz, 2H, CH2). LCMS (ESIþ): Calc. for [MþH]: 212.1; 214.1;
Found: 212.1; 214.1.
5.1.2.3. 1-(3-Chloropropyl)pyrrolidine 4c [27,35]. Purification by
column chromatography (DCM:MeOH(NH3), 99:1 (v/v)) was per-
formed and enabled to collect the product as a brown oil (703 mg,
30%). TLC: Rf 0.5 (DCM:MeOH(NH3), 9:1, v/v). 1H NMR (300 MHz,
CDCl3)
d
: 3.60 (t, J ¼ 6.9 Hz, 2H, CH2); 2.85 (t, J ¼ 7.0 Hz, 2H, CH2);
2.49 (m, 4H, 2 CH2); 1.98 (p, J ¼ 7.1 Hz, 2H, CH2); 1.78 (m, 4H, 2 CH2).
LCMS (ESIþ): Calc. for [MþH]: 148.1; 150.1; Found: 148.0; 150.0.
vents. Chemical shifts are reported as position
(
d
in ppm),
5.1.3. N-(3-Chloropropyl)isoindoline 5 [36]
multiplicity (s ¼ singlet, d ¼ doublet, t ¼ triplet, q ¼ quartet,
p ¼ pentet, dd ¼ double doublet, br ¼ broad and m ¼ multiplet),
coupling constant (J in Hz), relative integral and assignment. The
attributions of protons and carbons were achieved by analysis of 2D
experiments (COSY, HSQC and HMBC). Mass spectra were recorded
on a Varian triple quadrupole 1200 W mass spectrometer equipped
with a non-polar C18 TSK-gel Super ODS (4.6 ꢀ 50 mm) column,
using electrospray ionisation and a UV detector (diode array). The
purity of final compounds was verified by two types of high pres-
sure liquid chromatography (HPLC) columns: C18 Interchrom
UPTISPHERE and C4 Interchrom UPTISPHERE. Analytical HPLC was
performed on a Shimadzu LC-2010AHT system equipped with a UV
detector set at 254 nm and 215 nm. Compounds were dissolved in
A 1.0 g (8.4 mmol) amount of isoindoline was dissolved in 25 mL
of acetonitrile. After addition of potassium carbonate (2.3 g,
16.8 mmol), the resulting mixture was heated at 70 ꢁC. After
30 min, the solution was allowed to cool to room temperature. A
4.2 mL (47.2 mmol) amount of 1-bromo-3-chloropropane was
added and the reaction mixture stirred at room temperature for
24 h. The solvent was removed under reduced pressure and 60 mL
water added to the residue. The product was extracted with
3 ꢀ 30 mL dichloromethane. The combined organic fractions were
washed with water and dried over sodium sulphate. The solvent
was evaporated under reduced pressure. Purification by column
chromatography (DCM:MeOH(NH3), 99:1 (v/v)) was performed
and enabled collection of the product as a brown liquid (1.27 g,
77%). TLC: Rf 0.7 (DCM:MeOH(NH3), 9:1, v/v). 1H NMR (300 MHz,
50 mL methanol and 950 mL buffer A, and injected into the system.
The following eluent systems were used: buffer A (H2O/TFA,
100:0.1) and buffer B (CH3CN/H2O/TFA, 80:20:0.1). HPLC retention
times (HPLC tR) were obtained at a flow rate of 0.2 mL/min for
35 min using the following conditions: a gradient run from 100% of
buffer A over1 min, then to 100% of buffer B over the next 30 min.
CDCl3)
d
: 7.21 (s, 4H, Haro); 3.95 (s, 4H, 2 CH2); 3.69 (t, J ¼ 6.8 Hz, 2H,
CH2); 2.90 (t, J ¼ 7.1 Hz, 2H, CH2); 2.07 (p, J ¼ 7.0 Hz, 2H, CH2). 13
C
NMR (75 MHz, CDCl3) d: 140.0 (2 Caro); 126.7 (2 Caro); 122.3 (2 Caro);
59.1 (2 CH2); 53.0 (CH2); 43.1 (CH2); 31.9 (CH2). LCMS (ESIþ): Calc.
for [MþH]: 196.05; 198.07. Found: 195.95; 197.91.
5.1.2. General procedure for synthesis of compounds 4aec
5.1.4. General procedure for final compounds
A 1.00 g (16.9 mmol) amount of N-methylbenzylamine, N-
methyl-N-phenylethylamine, or pyrrolidine was dissolved in 25 mL
of DMF. After addition of potassium carbonate (4.6 g, 33.0 mmol),
the resulting mixture was heated at 70 ꢁC. After 30 min, the solu-
5.1.4.1. Procedure A. One equivalent of appropriate heterocyclic
derivative was dissolved in acetonitrile. Three equivalents of po-
tassium carbonate and 1.2 equivalents of appropriate 3-
chloropropan-1-amine derivative were added. The resulting
mixture was heated at 70 ꢁC until the disappearance of starting
material. The reaction was monitored by TLC. After 24e96 h, the
solvent was removed under reduced pressure and a 60 mL amount
of water added to the residue. The crude product was extracted
with dichloromethane. The combined organic fractions were
washed with water and dried over magnesium sulphate. Purifica-
tion by thick layer chromatography or column chromatography was
performed.
tion was allowed to cool to room temperature.
A 4.9 mL
(49.5 mmol) amount of 1-bromo-3-chloropropane was added and
the reacting mixture was stirred at room temperature for 24 h. The
solvent was removed under reduced pressure and 60 mL water
added to the residue. The product was extracted with 3 ꢀ 30 mL of
dichloromethane. The combined organic fractions were washed
with water and dried over sodium sulphate. The solvent was
evaporated under reduced pressure. Purification by column chro-
matography (DCM:MeOH(NH3), 99:1 (v/v)) was performed and
enabled to collect the product as an oil.
5.1.4.2. Procedure B. One equivalent of appropriate heterocyclic
derivative and 1.2 equivalents of appropriate 3-chloropropan-1-
5.1.2.1. N-Benzyl-3-chloro-N-methylpropan-1-amine
4
[34].
amine derivative were dissolved in a mixture of water:-
Purification by column chromatography (DCM) was performed and
enabled collection of the product as a colourless oil (3.2 g, 95%).
TLC: Rf 0.4 (DCM:MeOH(NH3), 99:1, v/v). 1H NMR (300 MHz, CDCl3)
acetonitrile (2:1, v/v). Three equivalents of sodium hydroxide were
added. The resulting mixture was heated at 50 ꢁC until the disap-
pearance of starting material. The reaction was monitored by TLC.
After 24e48 h, the product was extracted with ethyl acetate. The
combined organic fractions were washed with water and dried over
d: 7.40e7.22 (m, 5H, Haro); 3.65 (t, J ¼ 7.1 Hz, 2H, CH2); 3.52 (s, 2H,
CH2); 2.55 (t, J ¼ 7.0 Hz, 2H, CH2); 2.21 (s, 3H, CH3); 1.98 (p,

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203
magnesium sulphate. Purification by thick layer chromatography or
column chromatography was performed.
5.1.4.3.4. 2-[3-(N-Benzyl-N-methylamino)propyl]-1,2,3,4-
tetrahydrobenzofuro[3,2-c]pyridine 6d. Procedure A. The compound
was purified by column chromatography (DCM:MeOH(NH3), 99:1
(v/v)) and obtained as a yellowish oil (65 mg, 57%). TLC: Rf 0.4
5.1.4.3. Procedure C. The reaction was carried out under a nitrogen
atmosphere. One equivalent of appropriate heterocyclic derivative
was dissolved in anhydrous DMF. Four equivalents of sodium hy-
dride were added carefully and the resulting mixture was stirred at
room temperature for 30 min. Then 1.2 equivalents of appropriate
3-chloropropan-1-amine derivative were added. The reacting
mixture was stirred at room temperature until the disappearance of
starting material. The reaction was monitored by TLC. The reaction
was quenched with 50 mL of water and the crude product extracted
with ethyl acetate. The combined organic fractions were washed
with water and dried over magnesium sulphate. Purification by
column chromatography or thick layer chromatography was
performed.
(DCM:MeOH(NH3), 9:1, v/v). 1H NMR (300 MHz, CDCl3)
d: 7.46e7.24
(m, 7H, Haro); 7.23e7.18 (m, 2H, Haro); 3.64 (br t, J ¼ 2 Hz, 2H, H1);
3.54 (s, 2H, CH2); 2.94e2.85 (m, 4H, 2 CH2); 2.68 (t, J ¼ 7 Hz, 2H,
CH2); 2.50 (t, J ¼ 7 Hz, 2H, CH2); 2.25 (s, 3H, CH3); 1.87 (p, J ¼ 7 Hz,
2H, CH2). 13C NMR (75 MHz, CDCl3)
d: 154.7 (Caro); 152.1 (Caro);
129.3 (2 Caro); 128.4 (2 Caro); 127.4 (Caro); 127.1 (Caro); 123.2 (Caro);
122.4 (Caro); 118.2 (2 Caro); 111.6 (Caro); 111.0 (Caro); 62.0 (CH2); 55.3
(CH2); 55.2 (CH2); 50.3 (CH2); 48.7 (CH2); 41.9 (CH3); 25.1 (CH2);
24.2 (CH2). LCMS (ESIþ): Calc. for [MþH]: 335.2; Found: 335.1. HPLC
(C4, 35 min): tR 17.0 min, PHPLC 97%; HPLC (C18, 35 min): tR 15.0 min,
PHPLC > 99%.
5.1.4.3.5. 10-[3-(N-Benzyl-N-methylamino)propyl]phenoxazine
5.1.4.3.1. 2-[3-(N-benzyl-N-methylamino)propyl]-2,3,4,9-
tetrahydro-1H-pyrido[3,4-b] indole 6a. Procedure A. The compound
was purified by column chromatography (DCM: MeOH(NH3), 9:1, v/
v) and obtained as orange solid (202 mg, 35%). Melting point:
112 ꢁC. TLC: Rf 0.5 (DCM:MeOH(NH3), 8:2, v/v). 1H NMR (300 MHz,
6e. Procedure C. The compound was purified by column chroma-
tography (CycloHex:EtOAc, 3:2 (v/v) þ 0.1% MeOH(NH3)) and ob-
tained as a purple oil (219 mg, 58%). TLC: Rf 0.5 (CycloHex:EtOAc,
3:2, v/v). 1H NMR (300 MHz, CDCl3)
d: 7.39e7.29 (m, 5H, Haro);
6.81e6.50 (m, 8H, Haro); 3.58 (t, J ¼ 7 Hz, 2H, CH2); 3.54 (s, 2H, CH2);
DMSO-d6) d: 10.71 (br s, 1H, NH); 7.37e7.19 (m, 7H, Haro); 7.04e6.90
2.50 (t, J ¼ 7 Hz, 2H, CH2); 2.26 (s, 3H, CH3); 1.85 (p, J ¼ 7 Hz, 2H,
(m, 2H, Haro); 3.60 (br s, 2H, CH2); 3.48 (s, 2H, CH2); 2.74 (m, 2H,
CH2); 2.66 (m, 2H, CH2); 2.52 (t, J ¼ 7 Hz, 2H, CH2); 2.42 (t, J ¼ 7 Hz,
2H, CH2); 2.14 (s, 3H, CH3); 1.73 (p, J ¼ 7 Hz, 2H, CH2). 13C NMR
CH2). 13C NMR (75 MHz, CDCl3)
d: 145.0 (2 Caro); 133.2 (2 Caro);
129.7 (Caro); 128.7 (2 Caro); 123.7 (3 Caro); 121.0 (3 Caro); 115.5 (3
C
aro); 111.4 (2 Caro); 61.4 (CH2); 53.8 (CH2); 41.5 (CH2); 41.4 (CH3);
(75 MHz, DMSO-d6)
d
: 139.6 (Caro); 136.4 (Caro); 133.3 (Caro); 129.2
22.1 (CH2). LCMS (ESIþ): Calc. for [MþH]: 345.2; Found: 345.2. HPLC
(2 Caro); 128.6 (2 Caro); 127.3 (Caro); 127.1 (Caro); 120.7 (Caro); 118.7
(Caro); 117.8 (Caro); 111.4 (Caro); 106.9 (Caro); 62.0 (CH2); 55.8 (CH2);
55.3 (CH2); 51.3 (CH2); 50.6 (CH2); 42.3 (CH3); 25.1 (CH2); 21.7
(CH2). LCMS (ESIþ): Calc. for [MþH]: 334.2; Found: 334.0. HPLC (C4,
35 min): tR 10.2 min, PHPLC 97%; HPLC (C18, 35 min): tR 15.6 min,
(C4, 35 min): tR 14.0 min, PHPLC 98%; HPLC (C18, 35 min): tR 21.3 min,
PHPLC 98%.
5.1.4.3.6. 10-[3-(N-Benzyl-N-methylamino)propyl]phenothiazine
6f. Procedure C. The compound was purified by column chroma-
tography (CycloHex:EtOAc, 3:2 (v/v) þ 0.1% MeOH(NH3)) and ob-
tained as purple oil (238 mg, 66%). TLC: Rf 0.4 (CycloHex:EtOAc, 3:2,
PHPLC 97%.
5.1.4.3.2. 2-[3-(N-Benzyl-N-methylamino)propyl]-1-phenyl-
v/v). 1H NMR (300 MHz, CDCl3)
d: 7.30e7.13 (m, 9H, Haro);
2,3,4,9-tetrahydro-1H-pyrido[3,4-b] indole 6b. Procedure A. The
compound was purified by thick layer chromatography (cyclo-
hexane:ethyl acetate, 6:4 (v/v) þ 0.1% MeOH(NH3)) and obtained as
yellow solid (160 mg, 47%). TLC: Rf 0.5 (CycloHex:EtOAc,1:1, v/v). 1H
6.97e6.88 (m, 4H, Haro); 3.98 (t, J ¼ 7 Hz, 2H, CH2); 3.62 (s, 2H, CH2);
2.67 (t, J ¼ 7 Hz, 2H, CH2); 2.26 (s, 3H, CH3); 2.11 (p, 3J ¼ 7 Hz, 2H,
CH2). 13C NMR (75 MHz, CDCl3)
d: 145.1 (2 Caro); 129.5 (2 Caro); 129.2
(Caro); 128.5 (2 Caro); 127.7 (Caro); 127.5 (2 Caro); 127.3 (2 Caro); 125.4
(2 Caro); 122.6 (2 Caro); 115.7 (2 Caro); 61.6 (CH2); 54.1 (CH2); 44.8
(CH2); 41.5 (CH3); 23.9 (CH2). LCMS (ESIþ): Calc. for [MþH]: 361.1;
NMR (300 MHz, CDCl3) d: 7.54 (br s, 1H, NH); 7.42e7.23 (m, 11H,
H
aro); 7.22e7.05 (m, 3H, Haro); 4.60 (br t, J ¼ 2 Hz, 1H, CH); 3.46 (s,
2H, CH2); 3.35 (m, 1H, CH2); 2.93 (m, 2H, CH2); 2.79e2.62 (m, 2H,
CH2); 2.50e2.35 (m, 2H, CH2); 2.27 (m, 1H, CH2); 2.16 (s, 3H, CH3);
Found: 361.2. HPLC (C4, 35 min): tR 14.6 min, PHPLC 96%; HPLC (C18
35 min): tR 24.7 min, PHPLC 97%.
,
1.79 (p, J ¼ 7 Hz, 2H, CH2). 13C NMR (75 MHz, CDCl3)
d: 141.4 (Caro);
5.1.4.3.7. 3-[3-(N-Benzyl-N-methylamino)propyl]-2,3-dihydro-
2,5-dioxo-7-phenylcyclopenta-benzoxazole 6g. Procedure with
139.1 (Caro); 136.2 (Caro); 135.0 (Caro); 129.1 (2 Caro); 129.0 (2 Caro);
128.6 (2 Caro); 128.3 (2 Caro); 127.2 (Caro); 126.9 (Caro); 121.5 (Caro);
119.3 (Caro); 118.3 (Caro); 110.8 (Caro); 109.1 (Caro); 109.0 (Caro); 64.7
(CH2); 62.2 (CH2); 55.4 (CH2); 52.1 (CH2); 48.4 (CH2); 42.2 (CH3);
25.0 (CH2); 21.3 (CH2). LCMS (ESIþ): Calc. for [MþH]: 410.2; Found:
410.0. HPLC (C4, 35 min): tR 19.4 min, PHPLC 96%; HPLC (C18, 35 min):
tR 18.0 min, PHPLC 97%.
A
DMF. The compound was purified by column chromatography
(petroleum ether:ethyl acetate, 7:3 (v/v) þ 0.1% MeOH(NH3)) and
obtained as a beige solid (190 mg, 39%). Melting point: 93 ꢁC.TLC: Rf
0.2 (PE:EtOAc 7:3, v/v). 1H NMR (300 MHz, CDCl3)
d: 7.56 (s, 1H,
H
aro); 7.39e7.14 (m, 8H, Haro); 7.08 (dd, J ¼ 8 Hz, J ¼ 2 Hz, 2H, Haro);
6.85 (s, 1H, Haro); 4.54 (dd, J ¼ 8 Hz, J ¼ 3 Hz, 1H, CH2); 2.88 (m, 2H,
CH2); 3.38 (d, J ¼ 5 Hz, 2H, CH2); 3.26 (dd, J ¼ 19 Hz, J ¼ 4 Hz, 1H,
CH2); 2.71 (dd, J ¼ 19 Hz, J ¼ 4 Hz, 1H, CH2); 2.38 (t, J ¼ 7 Hz, 2H,
CH2); 2.02 (s, 3H, CH3); 1.91 (p, J ¼ 7 Hz, 2H, CH2). 13C NMR (75 MHz,
5.1.4.3.3. 2-[3-(N-Benzyl-N-methylamino)propyl]-2,3,4,5-
tetrahydro-1H-pyrido[4,3-b]indole 6c. Procedure A. The compound
was purified by column chromatography (DCM:MeOH(NH3), 95:5
to 94:6 (v/v)) and obtained as a yellow oil (15 mg, 3%). TLC: Rf 0.4
CDCl3) d: 204.0 (CO); 155.4 (Caro); 154.3 (CO); 143.4 (Caro); 142.8
(DCM: MeOH(NH3), 8:2, v/v). 1H NMR (300 MHz, CDCl3)
d
: 8.16
(Caro); 138.7 (Caro); 138.2 (Caro); 131.5 (Caro); 129.1 (2 Caro); 128.8 (2
Caro); 128.3 (2 Caro); 127.5 (2 Caro); 127.2 (Caro); 127.1 (Caro); 105.6
(Caro); 104.1 (Caro); 62.0 (CH2); 53.9 (CH2); 47.0 (CH2); 44.5 (CH2);
41.9 (CH3); 40.6 (CH2); 25.4 (CH2). LCMS (ESIþ): Calc. for [MþH]:
427.19; Found: 427.01. HPLC (C4, 35 min): tR 21.8 min, PHPLC 99%;
HPLC (C18, 35 min): tR 20.5 min, PHPLC 99%.
(br s, 1H, NH); 7.40 (m, 1H, Haro); 7.37e7.22 (m, 6H, Haro);
7.17e7.03 (m, 2H, Haro); 3.78 (br s, 2H, CH2); 3.55 (s, 2H, CH2);
2.97e2.81 (m, 4H, 2 CH2); 2.73 (t, J ¼ 7 Hz, 2H, CH2); 2.52 (t,
J ¼ 7 Hz, 2H, CH2); 2.26 (s, 3H, CH3); 1.93 (p, J ¼ 7 Hz, 2H, CH2).
13C NMR (75 MHz, CDCl3)
d: 138.7 (Caro); 136.1 (Caro); 131.9 (Caro);
129.2 (2 Caro); 128.3 (2 Caro); 127.2 (Caro); 126.0 (Caro); 121.3
(Caro); 119.3 (Caro); 117.5 (Caro); 110.7 (Caro); 108.0 (Caro); 62.3
(CH2); 55.9 (CH2); 55.4 (CH2); 50.7 (CH2); 49.6 (CH2); 42.2 (CH3);
25.0 (CH2); 23.5 (CH2). LCMS (ESIþ): Calc. for [MþH]: 334.2;
5.1.4.3.8. 3-[3-(N-Benzyl-N-methylamino)propyl]-2,3-dihydro-
2,5-dioxo-7-(4-chlorophenyl)
cyclopentabenzoxazole
6h.
Procedure A with DMF. The compound was purified by column
chromatography (DCM:MeOH(NH3), 98:2, v/v) and obtained as a
yellow solid (150 mg, 27%). Melting point: 146 ꢁC.TLC: Rf 0.5
Found: 334.0. HPLC (C4, 35 min): tR 5.3 min, PHPLC 97%; HPLC (C18
35 min): tR 14.9 min, PHPLC 97%.
,
(DCM:MeOH(NH3), 98:2, v/v). 1H NMR (300 MHz, CDCl3)
d: 7.57 (s,

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M. Donnier-Marechal et al. / European Journal of Medicinal Chemistry 89 (2015) 198e206
204
1H, Haro); 7.34e7.24 (m, 4H, Haro); 7.25e7.15 (m, 3H, Haro); 6.99 (d,
J ¼ 8 Hz, 2H, Haro); 6.82 (s, 1H, Haro); 4.50 (dd, J ¼ 8 Hz, J ¼ 4 Hz, 1H,
CH2); 3.88 (m, 2H, CH2); 3.38 (d, J ¼ 4 Hz, 2H, CH2); 3.25 (dd,
J ¼ 19 Hz, J ¼ 8 Hz, 1H, CH2); 2.65 (dd, J ¼ 19 Hz, J ¼ 4 Hz, 1H, CH2);
2.39 (t, J ¼ 7 Hz, 2H, CH2); 2.02 (s, 3H, CH3); 1.91 (p, J ¼ 7 Hz, 2H,
(DCM: MeOH(NH3), 9:1, v/v). 1H NMR (300 MHz, CDCl3)
d:
7.49e7.14 (m, 9H, Haro); 3.64 (br t, J ¼ 2 Hz, 2H, CH2); 3.00e2.73 (3,
8H, 4 CH2); 2.71e2.67 (m, 4H, 2 CH2); 2.51 (s, 3H, CH3); 1.96 (p,
J ¼ 7 Hz, 2H, CH2). 13C NMR (75 MHz, CDCl3)
d: 154.7 (Caro); 152.1
(Caro); 138.8 (Caro); 128.7 (2 Caro); 128.6 (2 Caro); 127.1 (Caro); 126.5
(Caro); 123.3 (Caro); 122.4 (Caro); 118.2 (Caro); 111.5 (Caro); 111.0
(Caro); 58.8 (CH2); 55.2 (CH2); 55.0 (CH2); 50.4 (CH2); 48.7 (CH2);
41.6 (CH3); 32.6 (CH2); 24.3 (CH2); 24.1 (CH2). LCMS (ESIþ): Calc. for
[MþH]: 349.2; Found: 349.1. HPLC (C4, 35 min): tR 10.9 min,
CH2). 13C NMR (75 MHz, CDCl3)
d: 203.5 (CO); 154.8 (Caro); 154.3
(CO); 142.9 (Caro); 141.9 (Caro); 138.7 (Caro); 138.5 (Caro); 133.0
(Caro); 131.5 (Caro); 129.2 (2 Caro); 128.9 (2 Caro); 128.8 (Caro); 128.3
(2 Caro); 127.1 (2 Caro); 105.5 (Caro); 104.2 (Caro); 62.0 (CH2); 53.9
(CH2); 46.8 (CH2); 43.9 (CH2); 41.8 (CH3); 40.7 (CH2); 25.5 (CH2).
LCMS (ESIþ): Calc. for [MþH]: 461.16; 463.16; Found: 461.01;
PHPLC > 99%; HPLC (C18, 35 min): tR 15.8 min, PHPLC > 99%.
5.1.4.3.13. 10-[3-(N-Methyl-N-phenylethylamino)propyl]phenox-
463.04. HPLC (C4, 35 min): tR 23.7 min, PHPLC 96%; HPLC (C18
35 min): tR 17.2 min, PHPLC 96%.
,
azine 7e. Procedure C. The compound was purified by column
chromatography (CycloHex:EtOAc, 3:2 (v/v) þ 0.1% MeOH(NH3))
and obtained as a yellow oil (200 mg, 51%). TLC: Rf 0.2 (Cyclo-
5.1.4.3.9. 2-[3-(N-Methyl-N-phenylethylamino)propyl]-2,3,4,9-
tetrahydro-1H-pyrido[3,4-b] indole 7a. Procedure A. The compound
was purified by column chromatography (DCM:MeOH(NH3), 99:1
to 97:3 (v/v)) and obtained as an orange oil (300 mg, 51%). TLC: Rf
Hex:EtOAc, 3:2, v/v). 1H NMR (300 MHz, CDCl3)
d
: 7.33e7.18 (m, 5H,
Haro); 6.82e6.74 (m, 2H, Haro); 6.68e6.61 (m, 4H, Haro); 6.54e6.50
(m, 2H, Haro); 3.53 (t, J ¼ 7 Hz, 2H, CH2); 2.83 (m, 2H, CH2); 2.65 (m,
0.6 (DCM:MeOH(NH3), 9:1, v/v). 1H NMR (300 MHz, CDCl3)
d: 8.11
2H, CH2); 2.54 (t, J ¼ 7 Hz, 2H, CH2); 2.35 (s, 3H, CH3); 1.81 (p,
(br s, 1H, NH); 7.48 (m, 1H, Haro); 7.34e7.27 (m, 3H, Haro); 7.25e7.18
(m, 3H, Haro); 7.17e7.08 (m, 2H, Haro); 3.63 (br s, 2H, CH2);
2.91e2.77 (m, 6H, 3 CH2); 2.73 (m, 2H, CH2); 2.62 (t, J ¼ 7 Hz, 2H,
CH2); 2.55 (t, 3J ¼ 7 Hz, 2H, CH2); 2.36 (s, 3H, CH3); 1.82 (p, J ¼ 7 Hz,
J ¼ 7 Hz, 2H, CH2). 13C NMR (75 MHz, CDCl3)
d: 145.0 (2 Caro); 140.0
(Caro); 133.3 (2 Caro); 128.7 (2 Caro); 128.5 (2 Caro); 126.2 (2 Caro);
123.7 (Caro); 120.8 (2 Caro); 115.3 (2 Caro); 111.42 (2 Caro); 59.3 (CH2);
54.7 (CH2); 42.0 (CH2); 41.7 (CH3); 33.6 (CH2); 22.7 (CH2). LCMS
(ESIþ): Calc. for [MþH]: 359.2; Found: 359.1. HPLC (C4, 35 min): tR
13.63 min, PHPLC 96%; HPLC (C18, 35 min): tR 24.0 min, PHPLC 96%.
5.1.4.3.14. 10-[3-(N-Methyl-N-phenylethylamino)propyl]pheno-
thiazine 7f. Procedure C. The compound was purified by column
chromatography (CycloHex:EtOAc, 3:2 (v/v) þ 0.1% MeOH(NH3))
and obtained as a brown oil (150 mg, 40%). TLC: Rf 0.2 (Cyclo-
2H, CH2). 13C NMR (75 MHz, CDCl3)
d: 140.5 (Caro); 136.2 (Caro);
131.9 (Caro); 128.7 (2 Caro); 128.4 (2 Caro); 127.2 (Caro); 126.1 (Caro);
121.3 (Caro); 119.3 (Caro); 118.0 (Caro); 110.7 (Caro); 108.3 (Caro); 59.9
(CH2); 55.9 (CH2); 55.6 (CH2); 51.3 (CH2); 50.4 (CH2); 42.2 (CH3);
33.7 (CH2); 25.3 (CH2); 21.3 (CH2). LCMS (ESIþ): Calc. for [MþH]:
348.2; Found: 348.0. HPLC (C4, 35 min): tR 9.3 min, PHPLC > 99%;
HPLC (C18, 35 min): tR 16.4 min, PHPLC > 99%.
Hex:EtOAc, 3:2, v/v). 1H NMR (300 MHz, CDCl3)
d: 77.32e7.10 (m,
5.1.4.3.10. 2-[3-(N-Methyl-N-phenylethylamino)propyl]-1-
9H, Haro); 6.97e6.84 (m, 4H, Haro); 3.94 (t, J ¼ 7 Hz, 2H, CH2);
phenyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]
indole
7b.
2.78e2.62 (m, 2H, CH2); 2.62e2.51 (m, 4H, 2 CH2); 2.35 (s, 3H, CH3);
Procedure A. The compound was purified by column chromatog-
raphy (DCM:MeOH(NH3), 98:2 to 96:4 (v/v)) and obtained as a
yellow oil (135 mg, 38%). TLC: Rf 0.6 (DCM:MeOH(NH3), 9:1, v/v). 1H
2.04 (p, J ¼ 7 Hz, 2H, CH2). 13C NMR (75 MHz, CDCl3)
d: 145.2 (2
C
aro); 139.6 (Caro); 128.7 (2 Caro); 128.5 (2 Caro); 127.6 (Caro); 127.3 (2
Caro); 126.2 (2 Caro); 125.4 (2 Caro); 122.6 (2 Caro); 115.8 (2 Caro); 59.1
NMR (300 MHz, CDCl3)
d
: 7.62 (m, 1H, Haro); 7.40e7.28 (m, 9H, Haro
(CH2); 54.6 (CH2); 44.9 (CH2); 42.0 (CH3); 32.8(CH2); 26.3 (CH2).
LCMS (ESIþ): Calc. for [MþH]: 375.1; Found: 374.9. HPLC (C4,
35 min): tR 14.4 min, PHPLC 97%; HPLC (C18, 35 min): tR 25.4 min,
and NH); 7.26e7.08 (m, 6H, Haro); 4.59 (br t, J ¼ 2 Hz, 1H, CH); 3.35
(m, 1H, CH2); 2.95 (m, 2H, CH2); 2.82e2.58 (m, 7H, CH and 3 CH2);
2.57e2.38 (m, 2H, CH2); 2.32 (s, 3H, CH3); 1.76 (p, J ¼ 7 Hz, 2H, CH2).
PHPLC 98%.
13C NMR (75 MHz, CDCl3)
d: 141.4 (Caro); 140.3 (Caro); 136.3 (Caro);
5.1.4.3.15. 2-[3-(Pyrrolidin-1-yl)propyl]-2,3,4,9-tetrahydro-1H-
134.9 (Caro); 129.1 (2 Caro); 128.7 (2 Caro); 128.6 (2 Caro); 128.4 (2
aro); 128.0 (Caro); 127.1 (Caro); 126.1 (Caro); 121.5 (Caro); 119.3 (Caro);
118.3 (Caro); 110.8 (Caro); 109.0 (Caro); 64.7 (CH2); 59.4 (CH2); 55.5
(CH2); 52.0 (CH2); 48.4 (CH2); 42.0 (CH3); 33.5 (CH2); 24.8 (CH2);
21.3 (CH2). LCMS (ESIþ): Calc. for [MþH]: 424.2; Found: 424.1. HPLC
(C4, 35 min): tR 11.0 min, PHPLC 97%; HPLC (C18, 35 min): tR 18.6 min,
pyrido[3,4,-b]indole 8a. Procedure A. The compound was purified
by column chromatography (DCM:MeOH(NH3), 96:4 (v/v)) and
obtained as a yellow oil (100 mg, 20%). TLC: Rf 0.6 (DCM:
C
MeOH(NH3), 9:1, v/v). 1H NMR (300 MHz, CDCl3)
d: 8.35 (br s, 1H,
NH); 7.49 (d, J ¼ 8 Hz, 1H, H8); 7.34 (d, 3J5-6 ¼ 8 Hz, 1H, H5); 7.17 (td,
J ¼ 8.2 Hz, J ¼ 1.2 Hz, 1H, H6); 7.11 (td, J ¼ 8.1 Hz, J ¼ 1.1 Hz, 1H, H7);
4.70 (br s, 2H, CH2); 4.25 (m, 2H, CH2); 3.85 (m, 2H, CH2); 2.85 (m,
2H, CH2); 2.77 (m, 6H, 3 CH2); 2.04 (p, J ¼ 7 Hz, 2H, CH2); 1.91e1.84
PHPLC 96%.
5.1.4.3.11. 2-[3-(N-Methyl-N-phenylethylamino)propyl]-2,3,4,5-
tetrahydro-1H-pyrido[4,3-b]indole 7c. Procedure B. The compound
was purified by thick layer chromatography (DCM:MeOH(NH3), 9:1
(v/v)) and obtained as a yellow oil (25 mg, 15%). TLC: Rf 0.3
(m, 4H, 2 CH2). 13C NMR (75 MHz, CDCl3)
d: 136.2 (Caro); 130.4
(Caro); 127.0 (Caro); 121.8 (Caro); 119.6 (Caro); 118.0 (Caro); 111.0 (Caro);
63.7 (CH2); 54.1 (CH2); 53.4 (2 CH2); 50.8 (CH2); 42.2 (CH2); 27.8
(CH2); 23.5 (2 CH2); 21.5 (CH2). LCMS (ESIþ): Calc. for [MþH]: 284.2;
(DCM:MeOH(NH3), 9:1, v/v). 1H NMR (300 MHz, CDCl3)
d: 7.96 (br s,
1H, NH); 7.41 (m, 1H, Haro); 7.33e7.18 (m, 6H, Haro); 7.17e7.05 (m,
2H, Haro); 3.74 (s, br, 2H, CH2); 2.95e2.79 (m, 6H, 3 CH2); 2.73e2.65
(m, 4H, 2 CH2); 2.58 (t, J ¼ 7.1 Hz, 2H, CH2); 2.39 (s, 3H, CH3); 1.89 (p,
Found: 284.1. HPLC (C4, 35 min): tR 10.6 min, PHPLC 96%; HPLC (C18
35 min): tR 15.7 min, PHPLC 96%.
,
5.1.4.3.16. 1-Phenyl-2-[3-(pyrrolidin-1-yl)propyl]-2,3,4,9-
tetrahydro-1H-pyrido[3,4-b] indole 8b. Procedure A. The compound
was purified by column chromatography (DCM:MeOH(NH3), 99:1
to 95:5 (v/v)) and obtained as a yellow oil (35 mg, 12%). TLC: Rf 0.6
J ¼ 7.3 Hz, 2H, CH2). 13C NMR (75 MHz, CDCl3)
d: 140.2 (Caro); 136.1
(Caro); 131.9 (Caro); 128.7 (2 Caro); 128.4 (2 Caro); 126.1 (Caro); 126.1
(Caro); 121.2 (Caro); 119.3 (Caro); 117.5 (Caro); 110.6 (Caro); 108.5
(Caro); 59.4 (CH2); 55.9 (CH2); 55.6 (CH2); 50.7 (CH2); 49.6 (CH2);
42.1 (CH3); 33.5 (CH2); 25.0 (CH2); 23.6 (CH2). LCMS (ESIþ): Calc. for
[MþH]: 348.2; Found: 348.0. HPLC (C4, 35 min): tR 8.3 min, PHPLC
98%; HPLC (C18, 35 min): tR 16.1 min, PHPLC 98%.
(DCM: MeOH(NH3), 8:2, v/v). 1H NMR (300 MHz, CDCl3)
d: 7.55 (m,
1H, Haro); 7.39e7.31 (m, 5H, Haro); 7.22e7.06 (m, 3H, Haro); 4.59 (br
t, J ¼ 2 Hz,1H, CH2); 3.33 (m, 1H, CH2); 2.99 (m, 2H, CH2); 2.82e2.77
(m, 2H, CH2); 2.62e2.51 (m, 5H, CH and 2 CH2); 2.45 (m, 1H, CH2);
2.33 (m, 1H, CH2); 1.90e1.67 (m, 6H, 3 CH2). 13C NMR (75 MHz,
CDCl3) d: 141.4 (Caro); 136.3 (Caro); 134.9 (Caro); 129.1 (2 Caro); 128.6
(2 Caro); 128.0 (Caro); 127.1 (Caro); 121.5 (Caro); 119.3 (Caro); 118.3
(Caro); 110.8 (Caro); 108.9 (Caro); 64.7 (CH2); 54.3 (CH2); 54.1 (2 CH2);
5.1.4.3.12. 2-[3-(N-Methyl-N-phenylethylamino)propyl]-1,2,3,4-
tetrahydrobenzofuro[3,2-c]pyridine 7d. Procedure A. The compound
was purified by thick layer chromatography (DCM:MeOH(NH3), 9:1
(v/v)) and obtained as a colourless oil (71 mg, 60%). TLC: Rf 0.5

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205
51.9 (CH2); 48.3 (CH2); 26.2 (CH2); 23.4 (2 CH2); 21.2 (CH2). LCMS
(ESIþ): Calc. for [MþH]:360.2; Found: 360.0. HPLC (C4, 35 min): tR
9.9 min, PHPLC 99%; HPLC (C18, 35 min): tR 15.4 min, PHPLC 98%.
5.1.4.3.17. 2-[3-(Isoindolin-2-yl)propyl]-2,3,4,9-tetrahydro-1H-
pyrido[3,4-b] indole 9a. Procedure A. The compound was purified
by column chromatography (DCM:MeOH(NH3), 99:1 to 96:4 (v/v))
and obtained as an orange solid (100 mg, 36%). Melting point:
197 ꢁC. TLC: Rf 0.5 (DCM: MeOH(NH3), 9:1, v/v). 1H NMR (300 MHz,
1H NMR (300 MHz, CDCl3)
d: 7.27 (s, 4H, Haro); 6.83e6.76 (m, 2H,
H
aro); 6.69e6.60 (m, 6H, Haro); 3.98 (s, 4H, 2 CH2); 3.68 (t, J ¼ 7 Hz,
2H, CH2); 2.85 (t, J ¼ 7 Hz, 2H, CH2); 1.94 (p, J ¼ 7 Hz, 2H, CH2). 13
C
NMR (75 MHz, CDCl3) d: 144.9 (2 Caro); 139.1 (2 Caro); 133.3 (2 Caro);
127.1 (2 Caro); 123.7 (2 Caro); 122.4 (2 Caro); 120.9 (2 Caro); 115.4 (2
C
aro); 111.4 (2 Caro); 59.2 (2 CH2); 53.8 (CH2); 41.6 (CH2); 24.2 (CH2).
LCMS (ESIþ): Calc. for [MþH]: 343.1; Found: 343.0. HPLC (C4,
35 min): tR 13.2 min, PHPLC 99%; HPLC (C18, 35 min): tR 23.9 min,
CDCl3)
d: 7.85 (br s, 1H, NH); 7.49 (m, 1H, Haro); 7.31 (m, 1H, Haro);
PHPLC > 99%.
7.21 (s, 4H, Haro); 7.17e7.06 (m, 2H, H6 and H7); 3.97 (s, 4H, 2 CH2);
3.73 (br t, J ¼ 2 Hz, 1H, CH); 2.95e2.81 (m, 6H, 3 CH2); 2.75 (t,
J ¼ 7 Hz, 2H, CH2); 1.94 (p, J ¼ 7 Hz, 2H, CH2). 13C NMR (75 MHz,
5.1.4.3.22. 10-[3-(Isoindolin-2-yl)propyl]phenothiazine
9f.
Procedure C. The compound was purified by column chromatog-
raphy (CycloHex:EtOAc, 3:2 (v/v) þ 0.1% MeOH(NH3)) and obtained
as a brown oil (199 mg, 56%). TLC: Rf 0.2 (CycloHex:EtOAc, 3:2, v/v).
CDCl3) d: 140.0 (2 Caro); 136.1 (Caro); 131.8 (Caro); 126.8 (2 Caro);
122.3 (2 Caro); 121.3 (Caro); 119.3 (Caro); 118.0 (Caro); 110.6 (Caro);
108.0 (Caro); 59.1 (2 CH2); 55.8 (CH2); 54.2 (CH2); 51.2 (CH2); 50.6
(CH2); 26.9 (CH2); 21.4 (CH2). LCMS (ESIþ): Calc. for [MþH]: 332.2;
1H NMR (300 MHz, CDCl3)
d: 7.23e7.10 (m, 8H, Haro); 6.98e6.88 (m,
4H, Haro); 4.02 (t, J ¼ 7 Hz, 2H, CH2); 3.93 (s, 4H, 2 CH2); 2.89 (t,
J ¼ 7 Hz, 2H, CH2); 2.09 (p, J ¼ 7 Hz, 2H, CH2). 13C NMR (75 MHz,
Found: 332.0. HPLC (C4, 35 min): tR 9.0 min, PHPLC > 99%; HPLC (C18
35 min): tR 15.2 min, PHPLC > 99%.
,
CDCl3) d: 145.3 (2 Caro); 139.6 (2 Caro); 127.5 (2 Caro); 127.3 (2 Caro);
126.9 (2 Caro); 125.3 (2 Caro); 122.5 (2 Caro); 122.3 (2 Caro); 115.7 (2
5.1.4.3.18. 2-[3-(Isoindolin-2-yl)propyl]-1-phenyl-2,3,4,9-
tetrahydro-1H-pyrido[3,4-b] indole 9b. Procedure A. The compound
was purified by thick layer chromatography (cyclohexane: ethyl
acetate, 1:1 (v/v) þ 0.1% MeOH(NH3)) and obtained as a yellow oil
(200 mg, 59%). TLC: Rf 0.5 (DCM: MeOH(NH3), 9:1, v/v). 1H NMR
C
aro); 59.2 (2 CH2); 53.6 (CH2); 45.2 (CH2); 26.3 (CH2). LCMS (ESIþ):
Calc. for [MþH]: 359.1; Found: 358.9. HPLC (C4, 35 min): tR
13.6 min, PHPLC 97%; HPLC (C18, 35 min): tR 24.0 min, PHPLC 96%.
5.2. In vitro testing
(300 MHz, CDCl3) d: 7.55 (m, 1H, Haro); 7.41e7.28 (m, 5H, Haro);
7.25e7.07 (m, 7H, Haro); 4.61 (br t, J ¼ 2 Hz, 1H, CH); 3.89 (s, 4H, 2
CH2); 3.39 (m, 1H, CH2); 2.95 (m, 2H, CH2); 2.82e2.67 (m, 3H, CH2);
2.65e2.46 (m, 2H, CH2); 1.84 (p, J ¼ 7 Hz, 2H, CH2). 13C NMR
5.2.1. Assay for binding to
s receptors
The binding assays were performed by CEREP (Poitiers,
s
France), according to Ganapathy et al. [32]. The s1 binding assay
was carried out by incubating Jurkat cell membranes (10e20 mg
protein per tube) with [3H](þ)-pentazocine (15 nM) and a range of
concentrations of test compounds, at 37 ꢁC for 2 h, in 5 mM Tris/HCl
buffer (pH ¼ 7.4). The s2 binding assay was performed by incu-
bating Jurkat cell membranes (10e20 mg protein per tube) with
(75 MHz, CDCl3) d: 141.5 (Caro); 139.9 (2 Caro); 136.3 (Caro); 135.0
(Caro); 129.2 (2 Caro); 128.7 (2 Caro); 128.0 (Caro); 127.1 (Caro); 126.7
(2 Caro); 122.2 (2 Caro); 121.5 (Caro); 119.3 (Caro); 118.3 (Caro); 110.8
(Caro); 109.0 (Caro); 64.7 (CH2); 59.0 (2 CH2); 54.1 (CH2); 51.9 (CH2);
48.4 (CH2); 26.7 (CH2); 21.3 (CH2). LCMS (ESIþ): Calc. for [MþH]:
408.2; Found: 408.0. HPLC (C4, 35 min): tR 11.9 min, PHPLC 98%; HPLC
(C18, 35 min): tR 17.0 min, PHPLC 96%.
[3H]-DTG (25 nM) in the presence of (þ)-pentazocine (1
mM) to
saturate s1 receptors, and a range of concentrations of test com-
pounds, at room temperature for 1 h in 5 mM TrisHCl buffer
(pH ¼ 7.4). The final assay volume was 0.5 mL. Binding was
terminated by rapid filtration through Whatman GF/B filters, which
were then washed with 5 ꢀ 1 mL ice-cold NaCl solution and
allowed to dry before bound radioactivity was measured using
liquid scintillation counting. Nonspecific binding was determined,
in both assays, under similar conditions, but in the presence of
5.1.4.3.19. 2-[3-(Isoindolin-2-yl)propyl]-2,3,4,5-tetrahydro-1H-
pyrido[4,3-b]indole 9c. Procedure B. The compound was purified by
thick layer chromatography (cyclohexane:ethyl acetate, 1:1 (v/
v) þ 0.1% MeOH(NH3)) and obtained as a yellow oil (41 mg, 26%).
TLC: Rf 0.4 (CycloHex:EtOAc, 1:1, v/v). 1H NMR (300 MHz, CDCl3)
d:
7.98 (br s, 1H, NH); 7.41 (m, 1H, Haro); 7.30 (m, 1H, Haro); 7.21 (s, 4H,
H
aro); 7.17e7.04 (m, 2H, Haro); 4.00 (s, 4H, Haro); 3.82 (br t, J ¼ 2 Hz,
2H, CH2); 3.02e2.77 (m, 8H, 4 CH2); 2.01 (p, J ¼ 7 Hz, 2H, CH2). 13
C
10 mM unlabelled haloperidol. Inhibition constants (Ki) were
calculated from the IC50 values according to the method of Cheng
and Prusoff (1973).
NMR (75 MHz, CDCl3)
d: 139.7 (2 Caro); 136.1 (Caro); 131.8 (Caro);
126.8 (2 Caro); 126.1 (Caro); 122.3 (2 Caro); 121.3 (Caro); 119.4 (Caro);
117.5 (Caro); 110.7 (Caro); 108.0 (Caro); 59.1 (2 CH2); 55.7 (CH2); 54.2
(CH2); 50.6 (CH2); 49.6 (CH2); 26.6 (CH2); 23.5 (CH2). LCMS (ESIþ):
Calc. for [MþH]: 332.2; Found: 332.0.
5.2.2. Cell culture and cytotoxicity assay
The human neuroblastoma cell line (SY5Y) was cultured in
DMEM (Dulbecco's Modified Eagle Medium) (Gibco) supplemented
5 .1. 4 . 3 . 2 0 . 2 - [ 3 - ( I s o i n d o l i n - 2 - yl ) p r o p yl ] - 1, 2 , 3 , 4 -
tetrahydrobenzofuro[3,2-c]pyridine 9d. Procedure A. The compound
was purified by thick layer chromatography (DCM:MeOH(NH3), 9:1
(v/v)) and obtained as a brown oil (67 mg, 58%). TLC: Rf 0.4 (DCM:
with 2 mM L-glutamine, 100 mg/ml streptomycin, 100 IU/mL peni-
cillin, 1 mM non-essential amino acids and 10% (v/v) heat-
inactivated foetal bovine serum (Sigma Aldrich), and grown at
37 ꢁC in a humidified incubator with 5% CO2.
MeOH(NH3), 9:1, v/v). 1H NMR (300 MHz, CDCl3)
d: 7.43e7.39 (m,
2H, Haro); 7.24e7.19 (m, 6H, Haro); 4.01 (s, 4H, Haro); 3.68 (br t,
J ¼ 2 Hz, 2H, CH2); 2.97e2.84 (m, 6H, 3 CH2); 2.78 (t, J ¼ 7 Hz, 2H,
Cells were seeded at 2000 cells per well onto 96-well plates in
DMEM medium. Cells were starved for 24 h to obtain synchronous
cultures, and were then incubated in culture medium that con-
tained various concentrations of test compounds, each dissolved in
less than 0.1% DMSO. After 72 h of incubation, cell growth was
estimated by the colorimetric MTT (thiazolyl blue tetrazolium
bromide) assay.
CH2); 1.97 (p, J ¼ 7 Hz, 2H, CH2). 13C NMR (75 MHz, CDCl3)
d: 154.7
(Caro); 152.1 (Caro); 139.6 (2 Caro); 127.2 (Caro); 126.9 (2 Caro); 123.2
(Caro); 122.4 (2 Caro); 122.3 (Caro); 118.2 (Caro); 111.6 (Caro); 111.0
(Caro); 59.1 (2 CH2); 55.4 (CH2); 54.2 (CH2); 50.4 (CH2); 48.7 (CH2);
26.9 (CH2); 24.3 (CH2). LCMS (ESIþ): Calc. for [MþH]: 333.2; Found:
333.2. HPLC (C4, 35 min): tR 16.7 min, PHPLC 97%; HPLC (C18, 35 min):
tR 14.7 min, PHPLC 96%.
Acknowledgements
5.1.4.3.21. 10-[3-(Isoindolin-2-yl)propyl]phenoxazine
9e.
Procedure C. The compound was purified by column chromatog-
raphy (CycloHex:EtOAc, 3:2 (v/v) þ 0.1% MeOH(NH3)) and obtained
as a brown oil (314 mg, 83%). TLC: Rf 0.5 (CycloHex:EtOAc, 3:2, v/v).
We express our thanks to Jiaqui He for her contribution in
ꢀ
organic synthesis and Amelie Barczyk for some in vitro experi-
ꢀ
ments. The 300 MHz NMR facilities were funded by the Region

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M. Donnier-Marechal et al. / European Journal of Medicinal Chemistry 89 (2015) 198e206
206
muscarinic and s₁ receptor agonist, in a nontransgenic mouse model of Alz-
ꢁ
Nord-Pas de Calais (France), the Ministere de la Jeunesse, de l'Ed-
ucation Nationale et de la Recherche (MJENR) and the Fonds
heimer's disease, Neuropsychopharmacol 38 (2013) 1706e1723.
[17] J. Charton, A.C. Gassiot, S. Girault-Mizzi, M.A. Debreu-Fontaine, P. Melnyk,
C. Sergheraert, Synthesis and pharmacological evaluation of Tic-hydantoin
ꢀ
ꢀ
ꢀ
Europeens de Developpement Regional (FEDER).
This work was supported by Lille 2 University, FRI “J'innove”
(A1112007 N), PRES Univ Lille Nord de France and Nord de France
SATT Grants. MDM is the recipient of a fellowship from Lille 2
University.
derivatives as selective
s1 ligands. Part 1, Bioorg. Med. Chem. Lett. 15
(2005) 4833e4837.
[18] A.C. Gassiot, J. Charton, S. Girault-Mizzi, P. Gilleron, M.A. Debreu-Fontaine,
C. Sergheraert, P. Melnyk, Synthesis and pharmacological evaluation of Tic-
hydantoin derivatives as selective
s1 ligands. Part 2, Bioorg. Med. Chem.
Lett. 15 (2005) 4828e4832.
[19] M. Toussaint, D. Mousset, C. Foulon, U. Jacquemard, C. Vaccher, P. Melnyk,
Sigma-1 ligands: Tic-hydantoin as a key pharmacophore, Eur. J. Med. Chem.
45 (2010) 256e263.
Appendix A. Supplementary data
[20] M. Toussaint, M.A. Debreu-Fontaine, T. Maurice, P. Melnyk, New synthesis of
tic-hydantoins sigma-1 ligands and pharmacological evaluation on cocaine-
induced stimulant effects, Med. Chem. 6 (2010) 355e373.
Supplementary data related to this article can be found at http://
dx.doi.org/10.1016/j.ejmech.2014.10.053.
[21] M. Toussaint, B. Delair, C. Foulon, N. Lempereur, C. Vaccher, T. Maurice,
P. Melnyk, Tic hydantoin sigma-1 agonist: pharmacological characterization
on cocaine-induced stimulant and appetitive effects, Eur. Neuro-
psychopharmacol. 19 (2009) 504e515.
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