Synthesis and antitumor activity of novel dithiocarbamate substituted chromones

Article abstract of DOI:10.1016/j.ejmech.2009.04.004

A series of chromone derivatives bearing diverse dithiocarbamate moieties were designed and synthesized via a three-component reaction protocol. Their in vitro antitumor activities were evaluated by MTT method against HCCLM-7, Hela, MDA-MB-435S, SW-480, H

Full text of DOI:10.1016/j.ejmech.2009.04.004

European Journal of Medicinal Chemistry 44 (2009) 3687–3696
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis and antitumor activity of novel dithiocarbamate substituted chromones
,
a,
**
Wei Huang a, Yu Ding a, Yan Miao a, Ming-Zhen Liu a, Yan Li b , Guang-Fu Yang
*
a Key Laboratory of Pesticide & Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, PR China
b Cancer Center of Wuhan University, Zhongnan Hospital of Wuhan University, Wuhan 430072, PR 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 chromone derivatives bearing diverse dithiocarbamate moieties were designed and synthe-
sized via a three-component reaction protocol. Their in vitro antitumor activities were evaluated by MTT
method against HCCLM-7, Hela, MDA-MB-435S, SW-480, Hep-2 and MCF-7. Two compounds (3-chloro-
4-oxo-4H-chromen-2-yl)methyl piperidine-1-carbodithioate (Iq) and (6-chloro-4-oxo-4H-chromen-3-
yl)methyl piperidine-1-carbodithioate (IIu), were identified as the most promising candidate due to their
high potency and broad-spectrum. Further flow-activated cell sorting analysis revealed that compounds
Iq and IIu arrest the cell cycle of SW-480 and MDA-MB-435s both in G2/M phase with dose-dependent
effect and might display apoptosis-inducing effect on these tumor cell lines.
Received 3 March 2009
Received in revised form
31 March 2009
Accepted 1 April 2009
Available online 9 April 2009
Keywords:
Flavonoids
Chromone
Ó 2009 Elsevier Masson SAS. All rights reserved.
Dithiocarbamate
Antitumor activity
1. Introduction
delay progression to AIDS. PDTC is a stable pyrrolidine derivative of
dithiocarbamates and an antioxidant. Previous studies have shown
As one of the most representative families of plant secondary
metabolites, flavonoids have been found to be associated with
a remarkable spectrum of biological activities [1–3]. In last decades,
medicinal chemists have paid great attentions on the isolation,
screening and structural modifications of new flavonoids. Most
interestingly, it has been found recently that some flavonoids dis-
played anticancer activity with novel mechanisms, such as carcin-
ogen inactivation, antiproliferation, cell cycle arrest, induction of
apoptosis and differentiation, inhibition of angiogenesis, anti-
oxidation and reversal of multidrug resistance [4]. To date, some
flavonoids with novel action mechanism have entered clinical
trials. For example, as shown in Fig. 1, flavopiridol was identified as
the first cyclin-dependent kinase inhibitor and entered Phase II
clinical trials [5].
that PDTC strongly inhibits replication of human rhinoviruses and
coxsackievirus myocarditis. PDTC also showed inhibitory ability
against murine colon adenocarcinoma bearing mice through the
inhibition of nuclear factor
kB in the tumor tissue [6,7]. Recently,
Dou et al. reported that the DSF–Cu complex showed inhibition of
proteasome activity and induction of apoptotic cell death [7].
Based on the above considerations, we proposed that chro-
mones bearing DTC moiety should display some interesting anti-
cancer activity. Therefore, we designed compounds I, II, III and IV
as shown in Fig. 2 with the aim to discover lead structure with
anticancer activity. Herein, we described the detailed synthetic
route, screening results and structure–activity relationships of
these designed compounds. Fortunately, two compounds with
promising broad-spectrum anticancer activity were identified.
Dithiocarbamate (DTC) derivatives are well known as organic
intermediates, rubber additive, additive of polluted water, vulca-
nizing agents and fungicides [6]. As shown in Fig.1, for example, DSF,
an irreversible inhibitor of aldehyde dehydrogenase, is one of the
two drugs approved by FDA for treatment of alcoholism [7]. Clinical
trials have shown the efficacy of DSF without toxicity. In clinical
trials DDTC was used in patients with HIV-1 infection and found to
2. Results and discussion
2.1. Chemistry
The synthetic route for compound I is outlined in Scheme 1.
According to the reported procedures [8,9], 2-methyl chromone 1
underwent bromination reaction with NBS to give the desired
2-bromomethyl chromone 2 in yields of 70–82%. Then, at the
presence of potassium phosphate as base [10], a subsequent three-
component reaction of the corresponding amine, carbon disulfide
and the intermediate 2 produced compound I in good yields
* Corresponding author.
** Corresponding author. Tel.: þ86 27 67867800; fax: þ86 27 67867141.
E-mail address: gfyang@mail.ccnu.edu.cn (G.-F. Yang).
0223-5234/$ – see front matter Ó 2009 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2009.04.004

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W. Huang et al. / European Journal of Medicinal Chemistry 44 (2009) 3687–3696
N
OH
O
S
H
S
S
HO
S
N
N
S
N
S
N
S
Cl
S
OH O
Flavopiridol
DSF
DDTC
PDTC
Fig. 1. Structures of flavopiridol and some representative dithiocarbamates.
(72–87%). As shown in Schemes 2 and 3, compounds II and III could
be easily prepared by the similar three-component reaction using
(mammary adenocarcinoma cell), were assayed by MTT method
[14] and the results expressed as IC50 were summarized in Table 1.
As shown in Table 1, the substituent on benzene ring of
compound I displayed remarkable effect on the antitumor activity.
For the derivatives bearing the same DTC moiety, introduction of
methoxyl group onto the position-7 always improved significantly
the antitumor activity, such as compounds Ia and If, Ic and Ig, Id
and Ih. However, introduction of Cl atom onto the position-7 did
not improve the activity. However, most interestingly, some
compounds bearing a chlorine atom at 3-position were found to
display good antitumor activities with broad-spectrum. For
3-choloromethyl chromone
3 and 3-bromochromone 4 as
substrate, respectively [11,12]. It should be noted that sodium
methoxide rather than potassium phosphate was used as base for
the three-component reaction of 3-bromochromone 4. Scheme 4
showed the synthetic route of compound IV. The bromination of
the starting material 5 with NBS afforded 6-bromomethyl chro-
mone 6 smoothly in a yield of 80% [9]. Then, three-component
reaction of the corresponding amine, carbon disulfide and inter-
mediate 6 produced the desired compound IV in good yields
(82–88%) when potassium phosphate was used as base.
The structures of all the target compounds were characterized
by elemental analyses, 1H NMR and EI-MS spectrum. In addition,
the crystal structures of Ic, IIs and IIIb were determined by X-ray
diffraction analyses [13]. As shown in Fig. 3, it can be found that the
special tropism of DTC side chain shows big difference. The dihedral
angles between the pyrone ring and DTC plane are 75.05ꢀ (Ic),
36.61ꢀ (IIs) and 78.67ꢀ (IIIb), respectively. The distance between
the centroids of pyrone ring and DTC unit are 4.46 Å (Ic), 4.65 Å (IIs)
and 3.91 Å (IIIb), respectively.
example, the IC50 values of compound Ip range from 0.6
Hep-2 to 2.3 M against MCF-7. The IC50 values of compound Iq are
0.77 M, 0.69 M, 0.94 M and 1.0 M against MDA-MB-435s, SW-
480, Hep-2 and MCF-7, respectively.
mM against
m
m
m
m
m
In most cases, compound II except for IIh and IIi displayed
higher antitumor activity than the corresponding compounds I. For
example, compound IIa and Ia (R ¼ H, D ¼ D2), IIb and Ib (R ¼ H,
D ¼ D3), IIc and Ic (R ¼ H, D ¼ D4), IId and Id (R ¼ H, D ¼ D5), IIj and
If (R ¼ 7-OMe, D ¼ D2), IIl and Ig (R ¼ 7-OMe, D ¼ D4), IIm and Ih
(R ¼ 7-OMe, D ¼ D5), IIt and Ik (R ¼ 7-OMe, D ¼ D3), IIu and Il
(R ¼ 7-OMe, D ¼ D4), IIv and Im (R ¼ 7-OMe, D ¼ D5). Most fortu-
nately, compound IIu displayed excellent antitumor activity with
broad-spectrum. Its IC50 values against the tested six cell lines are
2.2. Pharmacology
all under 1.0 mM, identified itself as the most promising candidate.
However, after the bridge CH2 group between the DTC moiety and
the chromone ring of compound II was removed, the antitumor
activity of the resulted compound III reduced significantly, indi-
cating that the flexibility of the DTC group might be a vital factor for
the activity. In addition, when the DTC moiety was moved onto
The in vitro antiproliferative activities of the synthesized
compounds I, II, III, and IV against six cancer cell lines, including
HCCLM-7 (hepatoma carcinoma cell), Hela (cervical carcinoma
cell), MDA-MB-435S (mammary adenocarcinoma cell), SW-480
(colon carcinoma cell), Hep-2 (laryngocarcinoma cell) and MCF-7
O
O
S
N
N
S
X
S
R
R
S
O
O
II
I
S
O
N
S
O
DTC
flavonoids
O
O
CO2Et
O
S
S
N
N
S
R
S
O
III
IV
Fig. 2. Design of the title compounds.

W. Huang et al. / European Journal of Medicinal Chemistry 44 (2009) 3687–3696
3689
S
O
O
O
O
O
S
N
Br
Amine, CS2
K3PO4
NBS
R
R
R
O
2a, R = H
Ia-r
1a, R = H
1b, R = 7-OMe
1c, R = 6-Cl
1d, R = 3-Cl
2b, R = 7-OMe
2c, R = 6-Cl
2d, R = 3-Cl
Scheme 1. Synthesis of 2-DTC chromones I.
O
O
Amine, CS2
N
S
Cl
K3PO4
R
R
S
O
O
IIa-v
3a, R = H
3b, R = 7-OMe
3c, R = 6-Me
3d, R = 6-Cl
Scheme 2. Synthesis of 3-DTC chromones II.
O
O
O
S
Amine, CS 2
K3PO4
N
S
R
R
Br
O
4a, R = H
IIIa-d
4b, R = 6-Cl
Scheme 3. Synthesis of 3-DTC chromones III.
O
CO2Et
O
CO2Et
O
CO2Et
NBS
Amine, CS2
K3PO4
S
N
S
O
O
6
Br
O
5
IVa-e
Scheme 4. Synthesis of 6-DTC chromones IV.
position-6 of chromone ring, the antitumor activities of the resul-
ted compound IV were reduced significantly.
performed [15]. The most promising compounds Iq and IIu were
tested against SW-480 and MDA-MB-435s cell lines respectively at
three different concentrations according the MTT assay. Fig. 4 and
Table 2 show the results after 48 h treatment for Iq and IIu,
respectively. As shown in Fig. 4 and Table 2, compound Iq arrest the
cell cycle of SW-480 in G2/M phase with dose-dependent effect,
In addition, from the results as shown in Table 1, we can also
conclude that the bulky DTC moieties, such as D7, D8, and D9, are not
favorable for the antitumor activity. In addition, although its
mechanism is not clear, D10 and D11 also seemed to display negative
effect on the antitumor activity. For example, compounds bearing
D10 and D11 moiety always displayed lower activity, such as
compounds Ie, IIi, IIr, and IVe.
raising the G2/M peak from 15.00% to 27.77% (0.5
mM), 27.96%
(0.7 M) and 39.27% (1.0 M) after 48 h treatment. Similarly,
m
m
compound IIu arrest the cell cycle of MDA-MB-435s in G2/M phase
To study the effect of the synthesized compounds on cell cycle
progression, flow-activated cell sorting (FACS) analysis was
with dose-dependent effect, raising the G2/M peak from 8.72% to
13.89% (0.12 mM), 20.62% (0.13 mM) and 24.87% (0.14 mM) after 48 h

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W. Huang et al. / European Journal of Medicinal Chemistry 44 (2009) 3687–3696
Fig. 3. X-ray crystal structures of Ic (A), IIs (B) and IIIb (C).
treatment. Based on the results of FACS analysis, we can conclude
compounds Iq and IIu might display apoptosis-inducing effect on
SW-480 and MDA-MB-435s cell lines, respectively.
two compounds (3-chloro-4-oxo-4H-chromen-2-yl)methyl piper-
idine-1-carbodithioate (Iq) and (6-chloro-4-oxo-4H-chromen-3-yl)-
methyl piperidine-1-carbodithioate (IIu), were identified as the
most promising candidate due to their high potency and broad-
spectrum. Further flow-activated cell sorting analysis revealed
that compounds Iq and IIu arrest respectively the cell cycle of
SW-480 and MDA-MB-435s in G2/M phase with dose-dependent
effect and might display apoptosis-inducing effect on these tumor
cell lines.
3. Conclusion
In conclusion, a series of chromones bearing diverse dithio-
carbamate moieties were designed and synthesized using a three-
component reaction protocol. Based on the result of MTT assay,

W. Huang et al. / European Journal of Medicinal Chemistry 44 (2009) 3687–3696
3691
Table 1
Antiproliferative activity of the DTC–chromone derivatives.
O
O
O
O
O
O
CO2Et
D
D
D
D
R
X
R
R
O
O
IV
III
I
II
CH(Ph-F-4)2
N
Bn
N
Ph
N
O
N
N
Cl
Cl
Ph
S
N
N
N
N
N
N
N
N
N
N
S
S
S
S
S
S
S
S
S
S
S
S
S
S
S
S
S
S
S
S
S
D11
D9
D7
D8
D10
D1
D2
D4
D5
D3
D6
a
N.O.
R
X
D
Antiproliferative activity (IC50, mM)
LM-7
Hela
–
23.1
>25
35.6
>25
>25
11.9
>25
8.7
>30
>40
>25
33.3
25.9
>40
0.7
9.8
4.5
>30
>25
2.3
MDA-MB-435s
SW-480
Hep-2
MCF-7
Ia
Ib
Ic
Id
Ie
If
Ig
Ih
Ii
Ij
Ik
Il
Im
In
Io
Ip
Iq
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
Cl
Cl
Cl
Cl
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
D2
D3
D4
D5
D11
D2
D4
D5
D1
D2
D3
D4
D5
D7
D2
D3
D4
D5
D2
D3
D4
D5
D6
D7
D8
D9
D10
D2
D3
D4
D5
D2
D3
D4
D5
D11
D2
D3
D4
D5
D3
D4
D3
D4
D2
D3
D4
D5
D10
>25
40.0
>25
25.2
>25
19.1
5.0
>40
>30
>25
22.2
3.7
22.1
1.5
3.4
23.5
>25
>25
3.0
>25
18.6
23.6
1.8
0.77
4.1
7.9
3.8
0.47
3.9
2.2
9.7
4.4
>40
23.4
13.7
24.4
25.1
7.2
13.6
9.1
12.0
>25
15.6
7.6
1.5
3.0
8.8
22.7
>40
1.6
20.6
15.9
20.0
0.6
0.94
9.0
3.0
3.6
0.39
4.4
2.4
16.7
2.9
9.6
>25
8.9
3.1
4.3
4.8
0.70
3.6
0.9
4.0
8.0
4.2
2.1
0.28
0.9
7.4
5.8
26.0
12.2
15.9
8.9
31.7
35.8
>25
21.0
>25
>25
10.0
>25
>25
>25
>25
>25
>25
>25
>40
2.3
7-OMe
7-OMe
7-OMe
6-Cl
6-Cl
6-Cl
6-Cl
6-Cl
6-Cl
H
H
H
H
H
H
H
H
H
2.3
3.1
11.0
8.8
>25
>25
>25
2.5
>25
21.2
18.1
1.4
0.69
1.8
6.8
2.3
1.2
>30
>25
13.8
>25
15.9
>40
1.9
3.3
1.0
Ir
19.2
21.9
6.0
11.1
>30
>25
1.7
IIa
IIb
IIc
IId
IIe
IIf
IIg
IIh
IIi
4.6
12.0
17.6
2.9
19.1
>25
>25
4.7
6.0
4.7
>40
6.3
3.4
–
5.9
4.0
5.9
7.1
7.9
>25
>25
–
22.9
>30
9.6
>30
>25
>40
34.1
>40
11.8
6.6
41.4
0.8
20.7
16.2
–
0.60
0.38
8.1
12.2
–
>25
>25
>25
>25
21.9
>25
>25
H
H
H
H
5.2
4.5
>40
>25
4.8
4.8
6.7
7.1
>25
5.9
>25
3.8
18.7
>25
7.8
>40
>25
1.8
IIj
IIk
IIl
7-OMe
7-OMe
7-OMe
7-OMe
6-Me
6-Me
6-Me
6-Me
6-Me
6-Cl
6-Cl
6-Cl
6-Cl
H
H
Cl
Cl
–
–
–
–
–
5.4
12.1
3.5
9.5
1.4
4.4
>25
6.1
23.3
>25
1.3
0.44
2.0
3.8
9.0
16.3
8.3
>25
6.6
5.8
>25
18.1
1.8
0.93
>40
0.24
1.9
1.7
5.5
6.7
>25
6.4
0.24
0.6
1.9
17.4
24.0
13.3
8.1
3.4
2.9
IIm
IIn
IIo
IIp
IIq
IIr
IIs
IIt
IIu
IIv
IIIa
IIIb
IIIc
IIId
IVa
IVb
IVc
IVd
IVe
5.1
14.3
14.5
>25
2.6
0.85
6.2
0.79
6.2
–
>25
16.3
>25
>25
>25
>25
>25
>25
>25
7.8
24.6
–
–
–
–
–
–
>25
>25
18.0
16.9
>25
>25
3.4
>25
–
>25
13.5
a
The IC50 values represent the concentration resulting in a 50% decrease in cell growth after 72 h incubation, which were the mean values of three repeated experiments.

3692
W. Huang et al. / European Journal of Medicinal Chemistry 44 (2009) 3687–3696
Fig. 4. Compounds Iq and IIu induced apoptosis in sw-480 cells and MDA-MS-435s respectively.
4. Experimental protocol
on silica gel with acetone/petroleum (15/85), giving the 2 in 70–
82% yield [9].
4.1. Chemistry
2-Methyl-4H-chromen-4-one (1a): Mp 70–72 ꢀC (Ref. [8d]:
72–73 ꢀC) 1H NMR (400 MHz, CDCl3):
d
¼ 2.33 (s, 3H, CH3), 6.87 (s,
4.1.1. General methods
1H, 3-CH), 7.47 (t, J ¼ 7.2 Hz, 1H, ArH), 7.55 (d, J ¼ 8.4 Hz, 1H, ArH),
7.71 (t, J ¼ 8.4 Hz, 1H, ArH), 7.92 (d, J ¼ 7.2 Hz, 1H, ArH); Reported:
2.36 (s, 3H, CH3), 6.90 (s, 1H, 3-CH), 7.46–7.74 (m, 3H, ArH),
7.75–7.95 (m, 1H, ArH).
1H NMR spectra were recorded at 400 MHz, in CDCl3 solution on
a Mercury-Plus400 spectrometer and chemical shifts were recor-
ded in parts per million (ppm) with TMS as the internal reference.
MS spectra were determined using a Tracems2000 organic mass
spectrometry, and signals were given in m/z. Melting points were
taken on a Buchi B-545 melting point apparatus. Element analysis
(EA) was measured on a Vario ELIII CHNSO elemental analyzer. All
commercially available solvents and reagents were used as
supplied by Acros Organics unless otherwise stated. The silica gel
(200–300 meshes) for flash column chromatography was from
Qingdao Marine Chemical Factory in China.
7-Methoxy-2-methyl-4H-chromen-4-one (1b): Mp 95–97 ꢀC;
1H NMR (400 MHz, CDCl3):
d
¼ 2.97 (s, 3H, CH3), 3.91 (s, 3H, OCH3),
6.19 (s, 1H, 3-CH), 6.74 (d, J ¼ 8.8 Hz, 1H, ArH), 7.21 (s, 1H, ArCH),
7.98 (d, J ¼ 8.8 Hz, 1H, ArH).
6-Chloro-2-methyl-4H-chromen-4-one (1c): Mp 116–118 ꢀC
(Ref. [8d]: 114–116 ꢀC); 1H NMR (400 MHz, CDCl3):
d
¼ 2.07 (s, 3H,
CH3), 5.30 (s, 1H, 3-CH), 7.39 (d, J ¼ 9.2 Hz, 1H, ArH), 7.47 (d,
J ¼ 9.2 Hz, 1H, ArH), 7.89 (s, 1H, ArCH); Reported: 2.10 (s, 3H, CH3),
5.30 (s, 1H, 3-CH), 7.41 (d, J ¼ 9.2 Hz,1H, ArH), 7.49 (d, J ¼ 9.2 Hz,1H,
ArH), 7.91 (s, 1H, ArCH).
4.1.2. General procedure for synthesis of I
The intermediate 2-methyl chromone
1
was synthesized
3-Chloro-2-methyl-4H-chromen-4-one (1d): Mp 115–117 ꢀC
according to the reported method [8]. The CCl4 solution (20 mL) of
1 (5 mmol), NBS (5 mmol) and AIBN (0.1 mmol) was irradiated
with 200 W lamp. After the reaction completed, the mixture was
cooled to room temperature and filtrated. The filtrate was
concentrated in vacuo. The crude product was chromatographed
(Ref. [8e]: 118 ꢀC); 1H NMR (400 MHz, CDCl3):
d
¼ 2.60 (s, 3H, CH3),
7.12 (t, J ¼ 8.0 Hz, 1H, ArH), 7.28 (d, J ¼ 8.4 Hz, 1H, ArH), 7.43 (t,
J ¼ 8.0 Hz, 1H, ArH), 8.20 (d, J ¼ 8.0 Hz, 1H, ArH); Reported: 2.58 (s,
3H, CH3), 7.07–7.45 (m, 3H, ArH), 8.21 (m, 1H, ArH).
2-(Bromomethyl)-4H-chromen-4-one (2a): Mp 98–99 ꢀC; 1H
NMR (400 MHz, CDCl3):
d
¼ 4.43 (s, 2H, CH2), 6.55 (s,1H, 3-CH), 7.36
(t, J ¼ 7.2 Hz,1H, ArH), 7.45 (d, J ¼ 8.4 Hz,1H, ArH), 7.66 (t, J ¼ 8.4 Hz,
1H, ArH), 8.14 (d, J ¼ 7.2 Hz, 1H, ArH); Ms (m/z) 239 (Mþ).
2-(Bromomethyl)-7-methoxy-4H-chromen-4-one (2b): Mp
Table 2
Effects of Iq and IIu on cell cycle progression.
N.O.
Iq to sw-480
IIu to MDA-MB-435s
Control 0.12 0.13
102–103 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 3.91 (s, 3H, CH3), 4.27
Control 0.5
m
M
0.7
m
M
1.0
m
M
m
M
mM
0.14 mM
(s, 2H, CH2), 6.43 (s, 1H, 3-CH), 6.89 (s, 1H, ArH), 6.99 (d, J ¼ 9.2 Hz,
1H, ArH), 8.08 (d, J ¼ 8.8 Hz, 1H, ArCH); Ms (m/z) 268 (Mþ).
2-(Bromomethyl)-6-chloro-4H-chromen-4-one (2c): Mp 110–
G0/G1 62.28
22.72
G2/M 15.00
39.39
37.84
27.77
36.84
35.20
27.96
26.51
34.22
39.27
83.62
7.62
8.72
60.82
25.29
13.89
58.68
20.70
20.62
41.57
33.56
24.87
S
112 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 4.54 (s, 2H, CH2), 6.68

W. Huang et al. / European Journal of Medicinal Chemistry 44 (2009) 3687–3696
3693
(s, 1H, 3-CH), 7.34 (d, J ¼ 9.2 Hz, 1H, ArH), 7.58 (d, J ¼ 9.2 Hz, 1H,
ArH), 7.73 (s, 1H, ArCH); Ms (m/z) 272 (Mþ).
(7-Methoxy-4-oxo-4H-chromen-2-yl)methyl
morpholine-4-
carbodithioate (Ih): Mp 141–143 ꢀC; 1H NMR (400 MHz, CDCl3):
3.79 (b, 4H, CH2), 3.94–3.98 (b, 5H, OMe & CH2), 4.35 (b, 2H, CH2),
4.61 (s, 2H, CH2), 6.46 (s, 1H, 3-CH), 6.86 (s, 1H, ArH), 6.96 (d,
J ¼ 9.2 Hz, 1H, ArH), 8.08 (d, J ¼ 8.8 Hz, 1H, ArCH); Ms (m/z) 351
(Mþ); Anal. calcd for C16H17NO4S2: C, 54.68; H, 4.88; N, 3.99. Found:
C, 54.57; H, 4.968; N, 4.12.
2-(Bromomethyl)-3-chloro-4H-chromen-4-one (2d): Mp 105–
107 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 4.82 (s, 2H, CH2), 7.41 (t,
J ¼ 8.0 Hz, 1H, ArH), 7.57 (d, J ¼ 8.4 Hz, 1H, ArH), 7.73 (t, J ¼ 8.0 Hz,
1H, ArH), 8.20 (d, J ¼ 8.0 Hz, 1H, ArH); Ms (m/z) 272 (Mþ).
To a solution of amine (1 mmol) in DMF (2 mL) was added
dropwise carbon disulfide (2 mmol) and anhydrous potassium
phosphate (1 mmol). The resulted mixture was stirred at room
temperature for 30 min. Then brominated chromone 2 (1 mmol)
was added by one-portion and stirring was continued. After
completion of the reaction (monitored by TLC), the mixture was
diluted with ice-cold water (20 mL) and the precipitate was filtered,
and recrystallized from ethanol to give the target compound I.
(4-Oxo-4H-chromen-2-yl)methyl diisopropylcarbamodithioate
(6-Chloro-4-oxo-4H-chromen-2-yl)methyl diethylcarbamodi
thioate (Ii): Mp 120–122 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.31 (m,
6H, CH3), 3.78 (m, 2H, CH2), 4.04 (m, 2H, CH2), 4.62 (s, 2H, CH2), 6.52
(s,1H, 3-CH), 7.42 (d, J ¼ 8.8 Hz,1H, ArH), 7.60 (d, J ¼ 8.8 Hz,1H, Het-
CH), 8.12–8.15 (m, 2H, ArCH); Ms (m/z) 341 (Mþ); Anal. calcd for
C15H16ClNO2S2: C, 52.70; H, 4.72; N, 4.10. Found: C, 52.70; H, 4.72;
N, 4.10.
(6-Chloro-4-oxo-4H-chromen-2-yl)methyl
diisopropylcarba-
(Ia): Mp 106–108 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.75 (b, 12H,
modithioate (Ij): Mp 104–106 ꢀC; 1H NMR (400 MHz, CDCl3):
CH3), 3.59 (b, 2H, iPr-CH), 4.64 (s, 2H, CH2), 6.51 (s, 1H, 3-CH), 7.36
(t, J ¼ 7.2 Hz,1H, ArH), 7.45 (d, J ¼ 8.4 Hz,1H, ArH), 7.66 (t, J ¼ 8.4 Hz,
1H, ArH), 8.14 (d, J ¼ 7.2 Hz, 1H, ArH); Ms (m/z) 335 (Mþ); Anal.
calcd for C17H21NO2S2: C, 60.86; H, 6.31; N, 4.18. Found: C, 60.71; H,
6.50; N, 4.29.
d
¼ 1.20–1.83 (b, 14H, i-Pr), 4.62 (s, 2H, CH2), 6.52 (s, 1H, 3-CH), 7.43
(d, J ¼ 8.8 Hz, 1H, ArH), 7.60 (d, J ¼ 8.8 Hz, 1H, Het-CH), 8.14 (s, 1H,
ArCH); Ms (m/z) 369 (Mþ); Anal. calcd for C17H20ClNO2S2: C, 55.19;
H, 5.45; N, 3.79. Found: C, 55.03; H, 5.61; N, 3.84.
(6-Chloro-4-oxo-4H-chromen-2-yl)methyl pyrrolidine-1-car-
(4-Oxo-4H-chromen-2-yl)methyl pyrrolidine-1-carbodithioate
bodithioate (Ik): Mp 120–122 ꢀC; 1H NMR (400 MHz, CDCl3):
(Ib): Mp 114–116 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 2.01 (m, 2H,
d
¼ 2.01 (q, 2H, CH2), 3.73 (q, 2H, CH2), 3.71 (d, J ¼ 6.8 Hz, 1H, CH2),
CH2), 2.13 (m, 2H, CH2), 3.71 (t, J ¼ 7.2 Hz, 2H, CH2), 3.96
(t, J ¼ 7.2 Hz, 2H, CH2), 4.64 (s, 2H, CH2), 6.53 (s, 1H, 3-CH), 7.39
(t, J ¼ 7.2 Hz,1H, ArH), 7.46 (d, J ¼ 8.4 Hz,1H, ArH), 7.66 (t, J ¼ 8.4 Hz,
1H, ArH), 8.18 (d, J ¼ 7.2 Hz, 1H, ArH); Ms (m/z) 305 (Mþ); Anal.
calcd for C15H15NO2S2: C, 58.99; H, 4.95; N, 4.59. Found: C, 58.70; H,
5.05; N, 4.39.
3.94 (d, J ¼ 6.8 Hz, 2H, CH2), 4.62 (s, 2H, CH2), 6.54 (s,1H, 3-CH), 7.42
(d, J ¼ 9.2 Hz, 1H, ArH), 7.60 (d, J ¼ 9.2 Hz, 1H, Het-CH), 8.13 (s, 1H,
ArCH); Ms (m/z) 339 (Mþ); Anal. calcd for C15H14ClNO2S2: C, 53.01;
H, 4.15; N, 4.12. Found: C, 52.89; H, 4.10; N, 4.22.
(6-Chloro-4-oxo-4H-chromen-2-yl)methyl piperidine-1-carbo-
dithioate (Il): Mp 131–133 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.73
(4-Oxo-4H-chromen-2-yl)methyl piperidine-1-carbodithioate
(b, 6H, CH2), 3.91 (b, 2H, CH2), 4.29 (b, 2H, CH2), 4.60 (s, 2H, CH2),
6.51 (s, 1H, 3-CH), 7.39 (d, J ¼ 8.8 Hz,1H, ArH), 7.60 (d, J ¼ 8.8 Hz,1H,
Het-CH), 8.14 (s, 1H, ArCH); Ms (m/z) 353 (Mþ); Anal. calcd for
C16H16ClNO2S2: C, 54.30; H, 4.56; N, 3.96. Found: C, 54.22; H, 4.71;
N, 3.87.
(Ic): Mp 120–122 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.76 (b, 6H,
CH2), 3.92 (m, 2H, CH2), 4.33 (m, 2H, CH2), 4.65 (s, 2H, CH2), 6.52
(s, 1H, 3-CH), 7.37 (t, J ¼ 7.2 Hz, 1H, ArH), 7.45 (d, J ¼ 8.4 Hz, 1H,
ArH), 7.66 (t, J ¼ 8.4 Hz, 1H, ArH), 8.18 (d, J ¼ 7.2 Hz, 1H, ArH); Ms
(m/z) 319 (Mþ); Anal. calcd for C16H17NO2S2: C, 60.16; H, 5.36; N,
4.38. Found: C, 60.02; H, 5.51; N, 4.43.
(6-Chloro-4-oxo-4H-chromen-2-yl)methyl morpholine-4-car-
bodithioate (Im): Mp 135–137 ꢀC; 1H NMR (400 MHz, CDCl3):
(4-Oxo-4H-chromen-2-yl)methyl morpholine-4-carbodithioate
d
¼ 3.80 (b, 4H, CH2), 3.98 (b, 2H, CH2), 4.35 (b, 2H, CH2), 4.64 (s, 2H,
(Id): Mp 131–134 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 3.79 (b, 4H,
CH2), 6.52 (s, 1H, 3-CH), 7.42 (d, J ¼ 10.4 Hz, 1H, ArH), 7.60
(d, J ¼ 10.4 Hz,1H, ArH), 8.14 (s, 1H, ArCH); Ms (m/z) 355 (Mþ); Anal.
calcd for C15H14ClNO3S2: C, 50.63; H, 3.97; N, 3.94. Found: C, 50.47;
H, 4.12; N, 4.05.
CH2), 3.96 (b, 2H, CH2), 4.35 (b, 2H, CH2), 4.64 (s, 2H, CH2), 6.52 (s,
1H, 3-CH), 7.38 (t, J ¼ 7.2 Hz, 1H, ArH), 7.46 (d, J ¼ 8.4 Hz, 1H, ArH),
7.66 (t, J ¼ 8.4 Hz, 1H, ArH), 8.18 (d, J ¼ 7.2 Hz, 1H, ArH); Ms (m/z)
321 (Mþ); Anal. calcd for C15H15NO3S2: C, 56.05; H, 4.70; N, 4.36.
Found: C, 56.12; H, 4.81; N, 4.40.
(6-Chloro-4-oxo-4H-chromen-2-yl)methyl 4-phenylpiperazine-
1-carbodithioate (In): Mp 163–165 ꢀC; 1H NMR (400 MHz, CDCl3):
(4-Oxo-4H-chromen-2-yl)methyl bis(2-chloroethyl)carbamo-
d
¼ 3.31 (b, 4H, CH2), 4.13 (b, 2H, CH2), 4.51 (b, 2H, CH2), 4.65 (s, 2H,
dithioate (Ie): Mp 142–144 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 3.26
CH2), 6.54 (s, 1H, 3-CH), 6.91–6.94 (m, 3H, ArH), 7.26–7.33 (m, 2H,
ArH), 7.43 (d, J ¼ 9.2 Hz, 1H, ArH), 7.61 (d, J ¼ 9.2 Hz, 1H, ArH), 8.14
(s, 1H, ArCH); Ms (m/z) 430 (Mþ); Anal. calcd for C21H19ClN2O2S2: C,
58.52; H, 4.44; N, 6.50. Found: C, 58.36; H, 4.23; N, 6.41.
(t, J ¼ 7.2 Hz, 2H, CH2), 3.57 (b, 4H, CH2), 3.73 (t, J ¼ 7.2 Hz, 2H,
CH2), 4.49 (s, 2H, CH2), 7.35 (d, J ¼ 8.4 Hz, 1H, ArH), 7.49 (d,
J ¼ 8.4 Hz, 1H, ArH), 7.78 (t, J ¼ 8.8 Hz, 1H, ArH), 7.94 (t, J ¼ 8.4 Hz,
1H, ArH), 8.19 (s, 1H, 3-CH); Ms (m/z) 375 (Mþ); Anal. calcd for
C15H15Cl2NO2S2: C, 47.87; H, 4.02; N, 3.72. Found: C, 47.60; H, 4.19;
N, 3.84.
(3-Chloro-4-oxo-4H-chromen-2-yl)methyl
diisopropylcarba-
modithioate (Io): Mp 112–114 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.77 (b, 12H, CH3), 3.648 (b, 2H, iPr-CH), 4.92 (s, 2H, CH2), 7.37
(7-Methoxy-4-oxo-4H-chromen-2-yl)methyl diisopropylcarba-
(t, J ¼ 8.0 Hz,1H, ArH), 7.47 (d, J ¼ 8.4 Hz,1H, ArH), 7.70 (t, J ¼ 8.0 Hz,
1H, ArH), 8.25 (d, J ¼ 8.0 Hz, 1H, ArH); Ms (m/z) 369 (Mþ); Anal.
calcd for C17H20ClNO2S2: C, 55.19; H, 5.45; N, 3.79. Found: C, 55.10;
H, 5.54; N, 3.68.
modithioate (If): Mp 121–123 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.22–1.66 (b, 14H, i-Pr), 3.91 (s, 3H, CH3), 4.60 (s, 2H, CH2), 4.62
(s, 2H, CH2), 6.43 (s, 1H, 3-CH), 6.86 (s, 1H, ArH), 6.96 (d, J ¼ 9.2 Hz,
1H, ArH), 8.08 (d, J ¼ 8.8 Hz, 1H, ArCH); Ms (m/z) 365 (Mþ); Anal.
calcd for C18H23NO3S2: C, 59.15; H, 6.34; N, 3.83. Found: C, 59.02; H,
6.40; N, 3.76.
(3-Chloro-4-oxo-4H-chromen-2-yl)methyl pyrrolidine-1-car-
bodithioate (Ip): Mp 126–128 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.93 (m, 2H, CH2), 2.05 (m, 2H, CH2), 3.62 (t, J ¼ 7.2 Hz, 2H, CH2),
(7-Methoxy-4-oxo-4H-chromen-2-yl)methyl piperidine-1-car-
3.92 (t, J ¼ 7.2 Hz, 2H, CH2), 4.92 (s, 2H, CH2), 7.37 (t, J ¼ 8.0 Hz, 1H,
ArH), 7.47 (d, J ¼ 8.4 Hz, 1H, ArH), 7.70 (t, J ¼ 8.0 Hz, 1H, ArH), 8.25
(d, J ¼ 8.0 Hz, 1H, ArH); Ms (m/z) 339 (Mþ); Anal. calcd for
C15H14ClNO2S2: C, 53.01; H, 4.15; N, 4.12. Found: C, 52.90; H, 4.02; N,
4.09.
bodithioate (Ig): Mp 130–132 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.73 (b, 6H, CH2), 3.90 (m, 5H, OMe & CH2), 4.30 (m, 2H, CH2),
4.64 (s, 2H, CH2), 6.45 (s, 1H, 3-CH), 6.86 (s, 1H, ArH), 6.96
(d, J ¼ 8.8 Hz, 1H, ArH), 8.07 (d, J ¼ 9.2 Hz, 1H, ArCH); Ms (m/z) 349
(Mþ); Anal. calcd for C17H19NO3S2: C, 58.43; H, 5.48; N, 4.01. Found:
C, 58.32; H, 5.60; N, 4.12.
(3-Chloro-4-oxo-4H-chromen-2-yl)methyl piperidine-1-carbo-
dithioate (Iq): Mp 140–142 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.74

3694
W. Huang et al. / European Journal of Medicinal Chemistry 44 (2009) 3687–3696
(b, 6H, CH2), 3.91 (b, 2H, CH2), 4.32 (b, 2H, CH2), 4.93 (s, 2H, CH2),
7.39 (t, J ¼ 8.0 Hz, 1H, ArH), 7.48 (d, J ¼ 8.4 Hz, 1H, ArH), 7.70 (t,
J ¼ 8.0 Hz,1H, ArH), 8.24 (d, J ¼ 8.0 Hz,1H, ArH); Ms (m/z) 353 (Mþ);
Anal. calcd for C16H16ClNO2S2: C, 54.30; H, 4.56; N, 3.96. Found: C,
54.17; H, 4.60; N, 3.90.
ArH), 8.40 (s, 1H, 2-CH); Ms (m/z) 321 (Mþ); Anal. calcd for
C15H15NO3S2: C, 56.05; H, 4.70; N, 4.36. Found: C, 55.95; H, 4.91; N,
4.50.
(4-Oxo-4H-chromen-3-yl)methyl 4-methylpiperazine-1-carbo-
dithioate (IIe): Mp 122–124 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 2.33
(3-Chloro-4-oxo-4H-chromen-2-yl)methyl morpholine-4-car-
(s, 3H, CH3), 2.50 (b, 4H, CH2), 4.36 (b, 2H, CH2), 4.48 (b, 2H, CH2),
4.52 (s, 2H, CH2), 6.54 (s, 1H, 3-CH), 7.27 (t, J ¼ 7.2 Hz, 1H, ArH), 7.39
(d, J ¼ 9.0 Hz, 1H, ArH), 7.66 (t, J ¼ 7.2 Hz, 1H, ArH), 8.24 (d,
J ¼ 9.0 Hz, 1H, ArH), 8.44 (s, 1H, ArCH); Ms (m/z) 334 (Mþ); Anal.
calcd for C16H18N2O2S2: C, 57.46; H, 5.42; N, 8.38. Found: C, 57.21; H,
5.60; N, 8.27.
bodithioate (Ir): Mp 141–143 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 3.75 (b, 4H, CH2), 3.94 (b, 2H, CH2), 4.32 (b, 2H, CH2), 4.97 (s, 2H,
CH2), 7.39 (t, J ¼ 8.0 Hz, 1H, ArH), 7.51 (d, J ¼ 8.4 Hz, 1H, ArH), 7.78 (t,
J ¼ 8.0 Hz,1H, ArH), 8.32 (d, J ¼ 8.0 Hz,1H, ArH); Ms (m/z) 355 (Mþ);
Anal. calcd for C15H14ClNO3S2: C, 50.63; H, 3.97; N, 3.94. Found: C,
50.49; H, 4.08; N, 3.85.
(4-Oxo-4H-chromen-3-yl)methyl 4-phenylpiperazine-1-carbo-
dithioate (IIf): Mp 151–153 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 3.31
4.1.3. General procedure for synthesis of II
(b, 4H, CH2), 4.13 (b, 2H, CH2), 4.51 (b, 2H, CH2), 4.65 (s, 2H, CH2),
6.54 (s, 1H, 3-CH), 6.91–6.94 (m, 3H, ArH), 7.26–7.33 (m, 2H, ArH),
7.43 (d, J ¼ 9.2 Hz, 1H, ArH), 7.61 (d, J ¼ 9.2 Hz, 1H, ArH), 8.14 (s, 1H,
ArCH); Ms (m/z) 396 (Mþ); Anal. calcd for C21H20N2O2S2: C, 63.61;
H, 5.08; N, 7.06. Found: C, 63.45; H, 5.31; N, 7.21.
The intermediate 3-choloromethyl chromone 3 was synthesized
according to the reported method [11].
3-(Chloromethyl)-4H-chromen-4-one (3a): Mp 105–107 ꢀC; 1H
NMR (400 MHz, CDCl3):
d
¼ 4.64 (s, 2H, CH2), 7.43 (d, J ¼ 8.0 Hz, 1H,
ArH), 7.57 (t, J ¼ 8.0 Hz, 1H, ArH), 7.76 (d, J ¼ 8.0 Hz, 1H, ArH), 8.08
(d, J ¼ 8.0 Hz, 1H, ArH), 8.14 (s, 1H, 2-CH); Ms (m/z) 194 (Mþ).
3-(Chloromethyl)-7-methoxy-4H-chromen-4-one (3b): Mp
(4-Oxo-4H-chromen-3-yl)methyl 4-benzylpiperazine-1-carbo-
dithioate (IIg): Mp 160–162 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 3.31
(b, 4H, CH2), 4.13 (b, 2H, CH2), 4.51 (b, 2H, CH2), 4.65 (s, 2H, CH2),
6.54 (s, 1H, 3-CH), 6.91–6.94 (m, 3H, ArH), 7.26–7.33 (m, 2H, ArH),
7.43 (d, J ¼ 9.2 Hz, 1H, ArH), 7.61 (d, J ¼ 9.2 Hz, 1H, ArH), 8.14 (s, 1H,
ArCH); Ms (m/z) 410 (Mþ); Anal. calcd for C22H22N2O2S2: C, 64.36;
H, 5.40; N, 6.82. Found: C, 64.21; H, 5.52; N, 6.76.
146–148 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 3.87 (s, 3H, CH3), 4.43
(s, 2H, CH2), 6.82 (s, 1H, 2-CH), 7.45 (d, J ¼ 8.8 Hz, 1H, ArH), 7.63 (d,
J ¼ 8.8 Hz, 1H, ArH), 8.10 (s, 1H, ArH); Ms (m/z) 224 (Mþ).
3-(Chloromethyl)-6-methyl-4H-chromen-4-one (3c): Mp 130–
132 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 2.42 (s, 3H, CH3), 4.42 (s, 2H,
(4-Oxo-4H-chromen-3-yl)methyl 4-(bis(4-fluorophenyl) methyl)
CH2), 7.50 (d, J ¼ 8.4 Hz, 1H, ArH), 7.64 (d, J ¼ 8.4 Hz, 1H, ArH), 8.01
piperazine-1-carbodithioate (IIh): Mp 175–177 ꢀC; 1H NMR
(s, 1H, ArH), 8.25 (s, 1H, 2-CH); Ms (m/z) 208 (Mþ).
(400 MHz, CDCl3):
d
¼ 3.31 (b, 4H, CH2), 4.13 (b, 2H, CH2), 4.51 (b,
6-Chloro-3-(chloromethyl)-4H-chromen-4-one (3d): Mp 159–
2H, CH2), 4.65 (s, 2H, CH2), 6.54 (s, 1H, 3-CH), 6.91–6.94 (m, 3H,
ArH), 7.26–7.33 (m, 2H, ArH), 7.43 (d, J ¼ 9.2 Hz, 1H, ArH), 7.61 (d,
J ¼ 9.2 Hz, 1H, ArH), 8.14 (s, 1H, ArCH); Ms (m/z) 522 (Mþ); Anal.
calcd for C28H24F2N2O2S2: C, 64.35; H, 4.63; N, 5.36. Found: C,
64.24; H, 4.45; N, 5.30.
161 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 4.53 (s, 2H, CH2), 7.45 (d,
J ¼ 8.8 Hz, 1H, ArH), 7.63 (d, J ¼ 8.8 Hz, 1H, ArH), 8.10 (s, 1H, ArH),
8.21 (s, 1H, 2-CH); Ms (m/z) 228 (Mþ).
To a solution of amine (1 mmol) in DMF (2 mL) was added
dropwise carbon disulfide (2 mmol) and anhydrous potassium
phosphate (1 mmol). The resulted mixture was stirred at room
temperature for 30 min. Then chlorinated chromone 3 (1 mmol)
was added by one-portion and stirring was continued. After
completion of the reaction (monitored by TLC), the mixture was
diluted with ice-cold water (20 mL) and the precipitate was filtered,
and recrystallized from ethanol to give the target compound II.
(4-Oxo-4H-chromen-3-yl)methyl diisopropylcarbamodithioate
(4-Oxo-4H-chromen-3-yl)methyl methyl(phenyl)carbamodi-
thioate (IIi): Mp 138–140 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 3.79
(s, 3H, CH3), 4.51 (s, 2H, CH2), 7.31 (d, J ¼ 6.8 Hz, 2H, ArH), 7.45–7.51
(m, 4H, ArH), 7.58 (d, J ¼ 8.0 Hz, 1H, ArH), 7.78 (t, J ¼ 7.2 Hz, 1H,
ArH), 8.17 (d, J ¼ 8.0 Hz,1H, ArH); Ms (m/z) 341 (Mþ); Anal. calcd for
C18H15NO2S2: C, 63.32; H, 4.43; N, 4.10. Found: C, 63.24; H, 4.61; N,
4.03.
(7-Methoxy-4-oxo-4H-chromen-3-yl)methyl diisopropylcarba-
(IIa): Mp 94–96 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.33 (b, 14H,
modithioate (IIj): Mp 96–98 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.73
CH2), 4.51 (s, 2H, CH2), 6.43 (d, J ¼ 9.6 Hz, 1H, Het-CH), 7.42 (t,
J ¼ 8.0 Hz, 1H, ArH), 7.45 (d, J ¼ 8.4 Hz, 1H, ArH), 7.65 (t, J ¼ 8.0 Hz,
1H, ArH), 8.24 (d, J ¼ 8.0 Hz, 1H, ArH), 8.43 (s, 1H, 2-CH); Ms (m/z)
335 (Mþ); Anal. calcd for C17H21NO2S2: C, 60.86; H, 6.31; N, 4.18.
Found: C, 60.69; H, 6.13; N, 4.24.
(b,12H, CH3), 3.58 (b, 2H, iPr-CH), 3.89 (s, 3H, CH3), 4.47 (s, 2H, CH2),
6.81 (s,1H, 2-CH), 6.96 (d, J ¼ 9.0 Hz,1H, ArH), 8.14 (d, J ¼ 9.6 Hz,1H,
ArH), 8.37 (s, 1H, ArH); Ms (m/z) 365 (Mþ); Anal. calcd for
C18H23NO3S2: C, 59.15; H, 6.34; N, 3.83. Found: C, 59.01; H, 6.40; N,
3.91.
(4-Oxo-4H-chromen-3-yl)methyl pyrrolidine-1-carbodithioate
(7-Methoxy-4-oxo-4H-chromen-3-yl)methyl pyrrolidine-1-car-
(IIb): Mp 103–105 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.93 (m, 2H,
bodithioate (IIk): Mp 106–108 ꢀC; 1H NMR (400 MHz, CDCl3):
d
CH2), 2.05 (m, 2H, CH2), 3.62 (t, J ¼ 7.2 Hz, 2H, CH2), 3.92
(t, J ¼ 7.2 Hz, 2H, CH2), 4.48 (s, 2H, CH2), 7.39–7.45 (m, 2H, ArH), 7.65
(t, J ¼ 8.0 Hz, 1H, ArH), 8.24 (d, J ¼ 8.0 Hz, 1H, ArH), 8.49 (s, 1H,
2-CH); Ms (m/z) 305 (Mþ); Anal. calcd for C15H15NO2S2: C, 58.99; H,
4.95; N, 4.59. Found: C, 58.80; H, 5.17; N, 4.63.
¼ 1.93 (m, 2H, CH2), 2.05 (m, 2H, CH2), 3.62 (t, J ¼ 7.2 Hz, 2H, CH2),
3.92 (t, J ¼ 7.2 Hz, 2H, CH2), 3.99 (s, 3H, CH3), 4.43 (s, 2H, CH2), 6.82
(s, 1H, 2-CH), 6.91 (d, J ¼ 9.0 Hz, 1H, ArH), 8.15 (d, J ¼ 9.6 Hz, 1H,
ArH), 8.35 (s, 1H, ArH); Ms (m/z) 335 (Mþ); Anal. calcd for
C16H17NO3S2: C, 57.29; H, 5.11; N, 4.18. Found: C, 57.31; H, 5.20; N,
4.30.
(4-Oxo-4H-chromen-3-yl)methyl piperidine-1-carbodithioate
(IIc): Mp 118–120 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 2.01 (b, 6H,
(7-Methoxy-4-oxo-4H-chromen-3-yl)methyl piperidine-1-car-
CH2), 2.71 (b, 2H, CH2), 4.18 (b, 2H, CH2), 4.64 (s, 2H, CH2), 7.21–7.25
(m, 2H, ArH), 7.57 (t, J ¼ 8.0 Hz, 1H, ArH), 8.20 (d, J ¼ 8.0 Hz, 1H,
ArH), 8.34 (s, 1H, 2-CH); Ms (m/z) 319 (Mþ); Anal. calcd for
C16H17NO2S2: C, 60.16; H, 5.36; N, 4.38. Found: C, 60.32; H, 5.21; N,
4.31.
bodithioate (IIl): Mp 124–126 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.73 (b, 6H, CH2), 3.90 (m, 5H, OMe & CH2), 4.30 (m, 2H, CH2),
4.56 (s, 2H, CH2), 6.89 (s, 1H, 2-CH), 7.03 (d, J ¼ 9.0 Hz, 1H, ArH), 8.19
(d, J ¼ 9.6 Hz, 1H, ArH), 8.51 (s, 1H, ArH); Ms (m/z) 349 (Mþ); Anal.
calcd for C17H19NO3S2: C, 58.43; H, 5.48; N, 4.01. Found: C, 58.54; H,
5.51; N, 4.05.
(4-Oxo-4H-chromen-3-yl)methyl morpholine-4-carbodithioate
(IId): Mp 131–134 ꢀC; 1H NMR (400 MHz, CDCl3):
CH2), 2.71 (b, 2H, CH2), 4.12 (b, 2H, CH2), 5.01 (s, 2H, CH2), 7.30–7.35
d
¼ 2.63 (b, 4H,
(7-Methoxy-4-oxo-4H-chromen-3-yl)methyl morpholine-4-
carbodithioate (IIm): Mp 136–138 ꢀC; 1H NMR (400 MHz, CDCl3):
3.72 (b, 4H, CH2), 3.88–3.93 (b, 5H, OMe & CH2), 4.32 (b, 2H, CH2),
(m, 2H, ArH), 7.61 (t, J ¼ 8.0 Hz, 1H, ArH), 8.26 (d, J ¼ 8.0 Hz, 1H,

W. Huang et al. / European Journal of Medicinal Chemistry 44 (2009) 3687–3696
3695
4.49 (s, 2H, CH2), 6.82 (s, 1H, 2-CH), 6.94 (d, J ¼ 9.0 Hz, 1H, ArH), 8.1
(d, J ¼ 8.4 Hz, 1H, ArH), 8.35 (s, 1H, ArH); Ms (m/z) 351 (Mþ); Anal.
calcd for C16H17NO4S2: C, 54.68; H, 4.88; N, 3.99. Found: C, 54.46; H,
4.95; N, 4.05.
(s, 2H, CH2), 7.41 (d, J ¼ 9.2 Hz, 1H, ArH), 7.61 (d, J ¼ 8.8 Hz, 1H, ArH),
8.19 (s, 1H, ArH), 8.44 (s, 1H, 2-CH); Ms (m/z) 355 (Mþ); Anal. calcd
for C15H14ClNO3S2: C, 50.63; H, 3.97; N, 3.94. Found: C, 50.50; H,
4.03; N, 4.11.
(6-Methyl-4-oxo-4H-chromen-3-yl)methyl diisopropylcarba-
modithioate (IIn): Mp 98–100 ꢀC; 1H NMR (400 MHz, CDCl3):
4.1.4. General procedure for synthesis of III
d
¼ 1.75 (b, 12H, CH3), 2.35 (s, 3H, CH3), 3.58 (b, 2H, iPr-CH), 4.48 (s,
The intermediate 3-bromo-chromones
according to the reported method [12].
4 was synthesized
2H, CH2), 7.34 (d, J ¼ 8.4 Hz, 1H, ArH), 7.46 (d, J ¼ 8.4 Hz, 1H, ArH),
8.01 (s, 1H, 2-CH), 8.42 (s, 1H, ArH); Ms (m/z) 349 (Mþ); Anal. calcd
for C18H23NO2S2: C, 61.86; H, 6.63; N, 4.01. Found: C, 61.72; H, 6.80;
N, 4.10.
3-Bromo-4H-chromen-4-one (4a): Mp 120–122 ꢀC; 1H NMR
(400 MHz, CDCl3):
d
¼ 7.39 (t, J ¼ 7.2 Hz, 1H, ArH), 7.52 (d, J ¼ 8.4 Hz,
1H, ArH), 7.73 (t, J ¼ 7.2 Hz, 1H, ArH), 8.15 (s, 1H, 2-CH), 8.31 (d,
(6-Methyl-4-oxo-4H-chromen-3-yl)methyl pyrrolidine-1-car-
J ¼ 7.6 Hz, 1H, ArH); Ms (m/z) 224 (Mþ).
bodithioate (IIo): Mp 110–113 ꢀC; 1H NMR (400 MHz, CDCl3):
3-Bromo-6-chloro-4H-chromen-4-one (4b): Mp 132–134 ꢀC; 1H
d
¼ 1.93 (m, 2H, CH2), 2.04 (m, 2H, CH2), 2.46 (s, 3H, CH3), 3.62 (t,
NMR (400 MHz, CDCl3):
d
¼ 7.48 (d, J ¼ 8.8 Hz, 1H, ArH), 7.66 (d,
J ¼ 7.2 Hz, 2H, CH2), 3.92 (t, J ¼ 7.2 Hz, 2H, CH2), 4.60 (s, 2H, CH2),
7.33 (d, J ¼ 8.4 Hz, 1H, ArH), 7.46 (d, J ¼ 8.4 Hz, 1H, ArH), 8.01 (s, 1H,
2-CH), 8.46 (s, 1H, ArH); Ms (m/z) 319 (Mþ); Anal. calcd for
C16H17NO2S2: C, 60.16; H, 5.36; N, 4.38. Found: C, 60.08; H, 5.50; N,
4.43.
J ¼ 8.8 Hz, 1H, ArH), 8.13 (s, 1H, ArH), 8.24 (s, 1H, 2-CH); Ms (m/z)
258 (Mþ).
To a solution of amine (1 mmol) in DMF (2 mL) was added
dropwise carbon disulfide (2 mmol) and anhydrous sodium
methoxide (1 mmol). The resulted mixture was stirred at room
temperature for 30 min. Then brominated chromone 4 (1 mmol)
was added by one-portion and stirring was continued. After
completion of the reaction (monitored by TLC), the mixture was
diluted with ice-cold water (20 mL) and the precipitate was filtered,
and recrystallized from ethanol to give the target compound III.
4-Oxo-4H-chromen-3-yl pyrrolidine-1-carbodithioate (IIIa):
(6-Methyl-4-oxo-4H-chromen-3-yl)methyl piperidine-1-carbo-
dithioate (IIp): Mp 121–123 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.74
(b, 6H, CH2), 2.45 (s, 3H, CH3), 3.91 (m, 2H, CH2), 4.31 (m, 2H, CH2),
4.56 (s, 2H, CH2), 7.32 (d, J ¼ 8.4 Hz, 1H, ArH), 7.45 (d, J ¼ 8.4 Hz, 1H,
ArH), 8.03 (s, 1H, 2-CH), 8.40 (s, 1H, ArH); Ms (m/z) 333 (Mþ); Anal.
calcd for C17H19NO2S2: C, 61.23; H, 5.74; N, 4.20. Found: C, 61.10; H,
5.90; N, 4.31.
(6-Methyl-4-oxo-4H-chromen-3-yl)methyl morpholine-4-car-
bodithioate (IIq): Mp 131–134 ꢀC; 1H NMR (400 MHz, CDCl3): 2.45
(s, 3H, CH3), 3.75 (b, 4H, CH2), 3.94 (b, 2H, CH2), 4.32 (b, 2H, CH2),
4.61 (s, 2H, CH2), 7.36 (d, J ¼ 8.4 Hz, 1H, ArH), 7.48 (d, J ¼ 8.4 Hz, 1H,
ArH), 8.07 (s, 1H, 2-CH), 8.49 (s, 1H, ArH); Ms (m/z) 335 (Mþ); Anal.
calcd for C16H17NO3S2: C, 57.29; H, 5.11; N, 4.18. Found: C, 57.13; H,
5.32; N, 4.30.
(6-Methyl-4-oxo-4H-chromen-3-yl)methyl bis(2-chloroethyl)-
carbamodithioate (IIr): Mp 146–148 ꢀC; 1H NMR (400 MHz, CDCl3):
3.26 (t, J ¼ 7.2 Hz, 2H, CH2), 3.70 (s, 3H, CH3), 3.72 (t, J ¼ 7.2 Hz, 2H,
CH2), 4.50 (s, 2H, CH2), 7.41–7.47 (m, 2H, ArH), 7.67 (d, J ¼ 8.4 Hz,1H,
ArH), 8.21–8.24 (m, 2H, ArH & 2-CH); Ms (m/z) 390 (Mþ); Anal.
calcd for C16H17Cl2NO2S2: C, 49.23; H, 4.39; N, 3.59. Found: C, 49.36;
H, 4.34; N, 3.61.
Mp 130–132 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 2.10 (m, 2H, CH2),
2.21 (m, 2H, CH2), 3.68 (t, J ¼ 7.2 Hz, 2H, CH2), 3.90 (t, J ¼ 7.2 Hz, 2H,
CH2), 7.39 (t, J ¼ 7.2 Hz, 1H, ArH), 7.52 (d, J ¼ 8.4 Hz, 1H, ArH), 7.73 (t,
J ¼ 7.2 Hz, 1H, ArH), 8.15 (s, 1H, 2-CH), 8.31 (d, J ¼ 7.6 Hz, 1H, ArH);
Ms (m/z) 291 (Mþ); Anal. calcd for C14H13NO2S2: C, 57.71; H, 4.50; N,
4.81. Found: C, 57.56; H, 4.70; N, 4.67.
4-Oxo-4H-chromen-3-yl piperidine-1-carbodithioate (IIIb): Mp
119–121 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.74 (b, 6H, CH2), 4.06
(b, 2H, CH2), 4.27 (b, 2H, CH2), 7.45 (t, J ¼ 7.2 Hz, 1H, ArH), 7.51 (d,
J ¼ 8.4 Hz, 1H, ArH), 7.71 (t, J ¼ 7.2 Hz, 1H, ArH), 8.13 (s, 1H, 2-CH),
8.27 (d, J ¼ 7.6 Hz, 1H, ArH); Ms (m/z) 305 (Mþ); Anal. calcd for
C15H15NO2S2: C, 58.99; H, 4.95; N, 4.59 6. Found: C, 58.86; H, 5.11; N,
4.47.
6-Chloro-4-oxo-4H-chromen-3-yl pyrrolidine-1-carbodithioate
(IIIc): Mp 123–125 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 2.01 (m, 2H,
(6-Chloro-4-oxo-4H-chromen-3-yl)methyl
diisopropylcarba-
CH2), 2.17 (m, 2H, CH2), 3.90 (m, 4H, CH2), 7.48 (d, J ¼ 8.8 Hz, 1H,
ArH), 7.66 (d, J ¼ 8.8 Hz, 1H, ArH), 8.13 (s, 1H, ArH), 8.24 (s, 1H,
2-CH); Ms (m/z) 325 (Mþ); Anal. calcd for C14H12ClNO2S2: C, 51.61;
H, 3.71; N, 4.30. Found: C, 51.45; H, 3.90; N, 4.42.
modithioate (IIs): Mp 110–112 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.32–1.58 (b, 12H, CH3), 4.08 (b, 1H, i-Pr-CH), 4.49 (s, 2H, CH2),
5.01 (b, 1H, i-Pr-CH), 7.41 (d, J ¼ 8.8 Hz, 1H, ArH), 7.60 (d, J ¼ 8.8 Hz,
1H, ArH), 8.20 (s, 1H, ArH), 8.44 (s, 1H, 2-CH); Ms (m/z) 369 (Mþ);
Anal. calcd for C17H20ClNO2S2: C, 55.19; H, 5.45; N, 3.79. Found: C,
54.98; H, 5.62; N, 3.65.
6-Chloro-4-oxo-4H-chromen-3-yl piperidine-1-carbodithioate
(IIId): Mp 130–133 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.78 (b, 6H,
CH2), 4.13 (b, 2H, CH2), 4.29 (b, 2H, CH2), 7.51 (d, J ¼ 8.8 Hz,1H, ArH),
7.68 (d, J ¼ 8.8 Hz, 1H, ArH), 8.13 (s, 1H, ArH), 8.27 (s, 1H, 2-CH); Ms
(m/z) 339 (Mþ); Anal. calcd for C15H14ClNO2S2: C, 53.01; H, 4.15; N,
4.12. Found: C, 53.12; H, 4.09; N, 4.23.
(6-Chloro-4-oxo-4H-chromen-3-yl)methyl pyrrolidine-1-car-
bodithioate (IIt): Mp 125–127 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.94–1.97 (m, 2H, CH2), 2.03–2.06 (m, 2H, CH2), 3.62
(d, J ¼ 6.8 Hz, 1H, CH2), 3.92 (d, J ¼ 6.8 Hz, 2H, CH2), 4.46 (s, 2H,
CH2), 7.40 (d, J ¼ 8.8 Hz, 1H, ArH), 7.59 (d, J ¼ 8.8 Hz, 1H, ArH), 8.19
(s, 1H, ArH), 8.50 (s, 1H, 2-CH); Ms (m/z) 339 (Mþ); Anal. calcd for
C15H14ClNO2S2: C, 53.01; H, 4.15; N, 4.12. Found: C, 52.78; H, 4.25; N,
4.09.
4.1.5. General procedure for synthesis of IV
The intermediate ethyl 6-methyl-4-oxo-4H-chromene-2-
carboxylate 5 was synthesized according to the reported method
[8,9]. The CCl4 solution (20 mL) of 5 (5 mmol), NBS (5 mmol) and
AIBN (0.1 mmol) was irradiated with 200 W lamp. After the reac-
tion completed, the mixture was cooled to room temperature and
filtrated. The filtrate was concentrated in vacuo. The crude product
was chromatographed on silica gel with acetone/petroleum
(15/85), giving 6 in 80% yield.
(6-Chloro-4-oxo-4H-chromen-3-yl)methyl piperidine-1-carbo-
dithioate (IIu): Mp 124–127 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.68
(b, 6H, CH2), 3.86 (b, 2H, CH2), 4.27 (b, 2H, CH2), 4.51 (s, 2H, CH2),
7.41 (d, J ¼ 9.2 Hz, 1H, ArH), 7.60 (d, J ¼ 9.2 Hz, 1H, ArH), 8.19 (s, 1H,
ArH), 8.47 (s, 1H, 2-CH); Ms (m/z) 353 (Mþ); Anal. calcd for
C16H16ClNO2S2: C, 54.30; H, 4.56; N, 3.96. Found: C, 54.42; H, 4.65;
N, 3.90.
Ethyl ethyl 6-methyl-4-oxo-4H-chromene-2-carboxylate (5):
Mp 137–139 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 1.45 (t, J ¼ 7.2 Hz,
(6-Chloro-4-oxo-4H-chromen-3-yl)methyl morpholine-4-car-
3H, CO2 CH23CH3), 2.00 (s, 3H, Ar-CH3), 4.36 (q, J ¼ 7.2 Hz, 2H,
CO2CH2), 7.16 (s, 1H, 3-CH), 7.53 (d, J ¼ 9.0 Hz, 1H, ArH), 7.88
(d, J ¼ 8.4 Hz, 1H, ArH), 8.20 (s, 1H, ArH); Ms (m/z) 232 (Mþ).
bodithioate (IIv): Mp 131–133 ꢀC; 1H NMR (400 MHz, CDCl3):
d
¼ 3.73 (b, 4H, CH2), 3.92 (b, 2H, CH2), 4.31 (b, 2H, CH2), 4.51

3696
W. Huang et al. / European Journal of Medicinal Chemistry 44 (2009) 3687–3696
Ethyl 6-(bromomethyl)-4-oxo-4H-chromene-2-carboxylate (6):
Mp 147–149 ꢀC; 1H NMR (400 MHz, CDCl3):
3H, CH3), 4.47 (q, J ¼ 7.2 Hz, 2H, CO2CH2), 4.72 (s, 2H, CH2), 7.11
(s, 1H, 3-CH), 7.56 (d, J ¼ 9.0 Hz, 1H, ArH), 7.85 (d, J ¼ 8.4 Hz, 1H,
ArH), 8.17 (s, 1H, ArH); Ms (m/z) 291 (Mþ).
Target compound IV can be synthesized from 6, amine and
carbon disulfide under the similar procedure to that of the
synthesis of I.
20 mL (0.5 mg/mL, final concentration) of MTT (Sigma, USA) was
d
¼ 1.44 (t, J ¼ 7.2 Hz,
added to each well and incubated for 4 h. MTT is converted to a blue
formazan product by mitochondrial succinate dehydrogenase. This
product was eluted from cells by addition of 150 mL of DMSO and
absorbance at 570 nm was determined by an ELISA using a NJ-2300
microplate spectrophotometer.
4.2.2. Flow activating cell sorting analysis (FACS) [15]
Ethyl 6-((diisopropylcarbamothioylthio)methyl)-4-oxo-4H-
The effect of compounds Iq and IIu on cancer cell cycle phase
distribution was assessed using flow cytometry. The cells were
grown to about 70% confluence in 6-well microplates and then
treated with Iq and IIu at different given concentrations. After 48 h,
cells were harvested by trypsinization, washed in PBS and fixed in
70% ice-cold (4 ꢀC) ethanol overnight. They were washed with PBS,
chromene-2-carboxylate (IVa): Mp 142–144 ꢀC; 1H NMR
(400 MHz, CDCl3):
d
¼ 1.34 (b, 14 H, CH2), 1.73 (b, 12 H, CH3), 3.58
(b, 2H, iPr-CH), 4.72 (s, 2H, CH2), 7.12 (s, 1H, 3-CH), 7.57 (d,
J ¼ 9.0 Hz, 1H, ArH), 7.86 (d, J ¼ 8.4 Hz, 1H, ArH, H), 8.17 (s, 1H,
ArH); Ms (m/z) 407 (Mþ); Anal. calcd for C20H25NO4S2: C, 58.94;
H, 6.18; N, 3.44. Found: C, 58.79; H, 6.34; N, 3.55.
incubated with RNase (100
m
g/mL final concentration) at 37 ꢀC for
g/mL final concentra-
Ethyl 4-oxo-6-((pyrrolidine-1-carbonothioylthio)methyl)-4H-
30 min, stained with propidium iodide (50
m
chromene-2-carboxylate (IVb): Mp 136–138 ꢀC; 1H NMR (400 MHz,
tion), and analyzed by flow cytometry (Beckman Coulter). For each
assay, three different experiments were performed in triplicate. The
results were statistically evaluated by the Student’s t-test. For all
the tests, the level of significance was set at P <0.05.
CDCl3):
d
¼ 1.44 (t, J ¼ 7.2 Hz, 3H, CH3), 1.98 (m, 2H, CH2), 2.09 (m,
2H, CH2), 3.65 (t, J ¼ 7.2 Hz, 2H, CH2), 3.96 (t, J ¼ 7.2 Hz, 2H, CH2),
4.47 (q, J ¼ 7.2 Hz, 2H, CO2CH2), 4.72 (s, 2H, CH2), 7.11 (s, 1H, 3-CH),
7.56 (d, J ¼ 9.0 Hz, 1H, ArH), 7.85 (d, J ¼ 8.4 Hz, 1H, ArH), 8.17 (s, 1H,
ArH); Ms (m/z) 377 (Mþ); Anal. calcd for C18H19NO4S2: C, 57.27; H,
5.07; N, 3.71. Found: C, 57.12; H, 5.00; N, 3.81.
Acknowledgments
Ethyl
chromene-2-carboxylate (IVc): Mp 151–153 ꢀC; 1H NMR (400 MHz,
CDCl3):
CH2), 4.16 (b, 2H, CH2), 4.72 (s, 2H, CH2), 7.11 (s, 1H, 3-CH), 7.54
(d, J ¼ 9.0 Hz, 1H, ArH), 7.87 (d, J ¼ 8.4 Hz, 1H, ArH, H), 8.14 (s, 1H,
ArH); Ms (m/z) 391 (Mþ); Anal. calcd for C19H21NO4S2: C, 58.29; H,
5.41; N, 3.58. Found: C, 58.09; H, 5.60; N, 3.63.
4-oxo-6-((piperidine-1-carbonothioylthio)methyl)-4H-
This work was supported by the Research Fund for the Doctoral
Program of Higher Education and in part by the National NSFC.
d
¼ 1.43 (t, J ¼ 7.2 Hz, 3H, CH3), 2.02 (b, 6H, CH2), 2.71 (b, 2H,
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CDCl3):
d
¼ 1.44 (t, J ¼ 7.2 Hz, 3H, CH3), 3.76 (b, 4H, CH2), 3.95 (b, 2H,
CH2), 4.35 (b, 2H, CH2), 4.47 (q, J ¼ 7.2 Hz, 2H, CO2CH2), 4.72 (s, 2H,
CH2), 7.11 (s, 1H, CH2), 7.57 (d, J ¼ 9.0 Hz, 1H, 3-CH), 7.82
(d, J ¼ 8.4 Hz,1H, ArH, H), 8.18 (s,1H, ArH); Ms (m/z) 393 (Mþ); Anal.
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H, 4.99; N, 3.60.
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(e) S.G. Jagadeesh, G.L.D. Krupadanam, G. Srimannarayana, Synth. Commun. 28
(1998) 3827–3833.
Ethyl 6-((methyl(phenyl)carbamothioylthio)methyl)-4-oxo-4H-
chromene-2-carboxylate (IVe): Mp 171–173 ꢀC; 1H NMR (400 MHz,
CDCl3):
d
¼ 1.43 (t, J ¼ 7.2 Hz, 3H, CH3), 1.56 (s, 3H, CH3), 2.05 (b, 6H,
CH2), 2.77 (b, 2H, CH2), 4.19 (b, 2H, CH2), 4.77 (s, 2H, CH2), 7.13
(s, 1H, 3-CH), 7.54–7.59 (m, 3H, ArH), 7.64–7.69 (m, 3H, ArH), 7.89
(d, J ¼ 8.4 Hz,1H, ArH, H), 8.18 (s, 1H, ArH); Ms (m/z) 413 (Mþ); Anal.
calcd for C21H19NO4S2: C, 61.00; H, 4.63; N, 3.39. Found: C, 61.08; H,
4.71; N, 3.48.
[9] H. Goker, D.W. Boykin, S. Yildiz, Bioorg. Med. Chem. 13 (2005) 1707–1714.
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[13] (a) Full crystallographic details have been deposited at the Cambridge Crys-
tallographic Data Center and allocated the deposition number CCDC 715986
(Ic), 715990 (IIs) and 715989 (IIIb). (b) G.M. Sheldrick, SHELXTL (Version 5.0),
4.2. Pharmacology
}
University of Gottingen, Germany, 2001;
(c) Bruker, SMART V5.628, SAINT V6.45. & SADABS, Bruker AXS Inc., Madison,
WI, USA, 2001.
[14] T. Mosmann, J. Immunol. Methods 65 (1983) 55–63.
[15] Z. Darzynkiewicz, E. Bednery, Methods Enzymol. 322 (2000) 18–39.
4.2.1. Cell proliferation by MTT assay [14]
Cells seeded in 96-well microplates at 8000 cells/well were
incubated with the test compounds for 72 h, respectively. Then,