Synthesis and biological evaluation of novel 1,6-diaryl pyridin-2(1H)-one analogs

Article abstract of DOI:10.1016/j.ejmech.2013.04.008

A series of 1,6-diaryl pyridin-2(1H)-one analogs were designed and synthesized via consecutive Chan-Lam coupling and Suzuki coupling. These novel compounds were evaluated for their antiproliferative activity against two tumor cell lines (SKOV-3 and HepG2). Compounds 1b, 1c, 1e and 1f displayed comparable in vitro cytotoxicity with taxol, while their in vivo antitumor activity was less effective than taxol. In addition, cell cycle assay indicated that these 1,6-diaryl pyridin-2(1H)-one compounds induced cell cycle arrest in the G1/M phase in the HepG2 cell line.

Full text of DOI:10.1016/j.ejmech.2013.04.008

European Journal of Medicinal Chemistry 64 (2013) 613e620
Contents lists available at SciVerse ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis and biological evaluation of novel 1,6-diaryl
pyridin-2(1H)-one analogs
,
b
c
d
Taijie Chen a, Yu Luo a , Youhong Hu , Bo Yang , Wei Lu
*
a Department of Chemistry, East China Normal University, 3663 North Zhongshan Road, Shanghai 200062, China
b Shanghai Institute of Materia Medica, Chinese Academy of Science, 555 Zu Chong Zhi Road, Shanghai 201203, China
c Institute of Pharmacology and Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, Zhejiang, China
d Institute of Drug Discovery and Development, East China Normal University, 3663 North Zhongshan Road, Shanghai 200062, 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 1,6-diaryl pyridin-2(1H)-one analogs were designed and synthesized via consecutive Chan
eLam coupling and Suzuki coupling. These novel compounds were evaluated for their antiproliferative
activity against two tumor cell lines (SKOV-3 and HepG2). Compounds 1b, 1c, 1e and 1f displayed
comparable in vitro cytotoxicity with taxol, while their in vivo antitumor activity was less effective than
taxol. In addition, cell cycle assay indicated that these 1,6-diaryl pyridin-2(1H)-one compounds induced
cell cycle arrest in the G1/M phase in the HepG2 cell line.
Received 3 February 2013
Received in revised form
19 March 2013
Accepted 2 April 2013
Available online 11 April 2013
Ó 2013 Elsevier Masson SAS. All rights reserved.
Keywords:
Pyridin-2(1H)-one
ChaneLam coupling reaction
Cytotoxicity
Antitumor activity
Cell cycle
1. Introduction
Thus, it was postulated that these novel pyridone analogs might
exhibit antitumor activity like Combretastatin A-4, which shows
According to the most recent cancer statistics [1] in the United
States and other countries, the burden of malignancies continues to
increase worldwide, so that there is an urgent need for new
effective, safe, and affordable drugs. During the past few years, the
pyridin-2(1H)-one ring system derivatives have been considered as
some of the most promising antitumor [2e5] agents (Fig. 1). In
addition, pyridones have been recognized as privileged core
structures [6,7] and existed in many bioactive natural products [8]
and various pharmaceutical agents [9,10]. Existing studies have
proved their wide range of biological activities [11e16] such as
cardiotonic [17], antibacterial [18], antihepatitis B virus [19].
Although a significant volume of literature existed on various
multi-substituted pyridin-2(1H)-one derivatives, few literatures
focused on the synthesis and biological profiles of 1,6-diaryl-
substituted pyridin-2(1H)-ones. By analyzing the structure of these
pyridones, it could be found that they were similar to Com-
bretastatin A-4 and its analogs [20e22] (Fig. 2), as these structures
were all characterized by two cis-conformational aryl-substituents.
potent antitumor activity against a variety of human cancer cells.
Herein, we report a series of novel pyridin-2(1H)-one analogs
synthesized via ChaneLam coupling reaction and Suzuki coupling
reaction. The synthesized compounds were then evaluated for their
in vitro and in vivo antitumor activity, and subjected to cell cycle
arrest analysis.
2. Results and discussion
2.1. Chemistry
Since Chan, Evans and Lam developed a new copper-catalyzed
N-arylation methodology from amines and arylboronic acids, this
method has soon proved to be a powerful and attractive synthetic
tool to construct carboneheteroatom bond and widely used in the
synthesis of many biologically active compounds. In addition,
palladium-catalyzed Suzuki coupling is also a widely used meth-
odology to generate CeC bond from halolides and arylboronic
acids. Consequently, 6-bromopyridin-2(1H)-one with both a CeBr
bond and a NeH bond, was selected as a suitable starting material.
However, to the best of our knowledge, few literatures exist on the
N-arylation of 6-bromopyridin-2(1H)-one via ChaneLam reaction.
* Corresponding author. Tel./fax: þ86 21 62602475.
E-mail addresses: b97131113@yahoo.com.cn, yluo@chem.ecnu.edu.cn (Y. Luo).
0223-5234/$ e see front matter Ó 2013 Elsevier Masson SAS. All rights reserved.
http://dx.doi.org/10.1016/j.ejmech.2013.04.008

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T. Chen et al. / European Journal of Medicinal Chemistry 64 (2013) 613e620
Fig. 1. Structures of some important pyridone derivatives.
Kossen et al. [23] reported a preparation of related N-arylated 6-
bromopyridin-2(1H)-one products using classical ChaneLam re-
action conditions. However, the designed compounds were not
obtained following Kossen’s protocol (Table 1, entry 1). Thus, we
had to evaluate the reaction conditions and selected 6-
bromopyridin-2(1H)-one and 3,4,5-trimethoxyphenylboronic acid
as the test substrates.
2.2. Biology
2.2.1. Cytotoxicity studies
The two subseries of pyridin-2(1H)-one derivatives were eval-
uated in vitro for their antiproliferative activity against human
ovarian carcinoma cell line (SKOV-3) and human hepatoma cell line
(HepG2) using taxol as the reference compound.
As depicted in Table 1, to screen for suitable reaction conditions,
different catalysts, bases, solvents and additives were surveyed. It
was found that the additives were crucial to this coupling. Although
CsF [24] was reported to increase the yield of the coupling, it was
basically of no use in this reaction (Table 1, entries 3e6). Fortunately,
the yields were dramatically increased (Table 1, entry 8), when
pyridine-N-oxide [25] was used as an additive. Thus, the optimal
conditions for the N-arylationwere determined as following: 10 mol
% of Cu(OAc)2 as the catalyst, pyridine (2 eq) as the base, pyridine-N-
oxide (1.1 eq) and 4 ÅMS as the additives and stirring at room
temperature for 24 h. Subsequently, the intermediate 4 was reacted
with a variety of arylboronic acids under traditional Suzuki coupling
conditions to afford 1aem in good yields (Scheme 1). Thereafter, the
second subseries 2aej was prepared in a reversed coupling
sequence. 6-Bromopyridin-2(1H)-one was first subjected to the
Suzuki coupling with 3,4,5-trimethoxyphenylboronic acid to give
compound 5 as a common intermediate. Using the screened Chane
Lam coupling conditions, compound 5 was coupled with arylbor-
onic acids to produce 2aej in moderate yields.
As shown in Table 2, the subseries 1aen generally displayed
more potent antiproliferative activity than the subseries 2aej. The
most potent analogs were compound 1b, 1c, 1e and 1f. Compound
1b displayed very potent antiproliferative activity (IC50 <0.01 mM),
while blocking the hydroxyl (1a, 1d and 1k) led to a loss of anti-
proliferative activity. Comparison of bioactivity between 1b and 1c
revealed that these two compounds were equipotent to each other
in the two cell lines (SKOV-3 and HepG2) (IC50 <0.01 mM), while
replacement of 4-methoxyl group (compound 1l) led to a signifi-
cant loss of activity. Therefore, 4-methoxyl was of benefit to its
antiproliferative activity. The 3-methyl (1j) and 3-F (1g) were
detrimental to the activity. Introduction of piperonyl (1e)
enhanced antiproliferative activity with a selectivity in the two
cell lines [IC50 0.064
mM (SKOV-3 cells) and <0.01 mM (HepG2
cells)]. In addition, 4-NO2 compound (1f) also exhibited good
antiproliferative activity with a selectivity in the two cell lines
[IC50 0.201 mM (SKOV-3 cells) and <0.01 mM (HepG2 cells)], while
introduction other groups (1h, 1i and 1m) did not increase the
activity in the two cell lines.
Fig. 2. Structures of CA-4 and target compounds.

T. Chen et al. / European Journal of Medicinal Chemistry 64 (2013) 613e620
615
Table 1
Optimization of the ChaneLam reaction.
Entry
Catalyst
Base
Additive
Solvent
Temperature
Yielda
1
2
3
4
5
6
7
8
Cu(OAc)2∙H2Ob
Py
Py
Py
Py
Py
NaOH
Py
e
CH2Cl2
CH2Cl2
CH2Cl2
PhMe
PhMe
PhMe
RT
RT
RT
80 ꢀC
80 ꢀC
80 ꢀC
RT
/
Cu(OAc)2∙H2Ob
4 ÅMS
Trace
/
2%
17%
/
b
Cu(OAc)2
CsF (1 eq)/4 ÅMS
CsF (1 eq)/4 ÅMS
CsF (1 eq)/4 ÅMS
CsF (1 eq)/4 ÅMS
Py-NO (1.1 eq)/4 ÅMS
Py-NO (1.1 eq)/ÅMS
b
Cu(OAc)2
Cu(OAc)2 (2 eq)
Cu(OAc)2 (2 eq)
Cu(OAc)2 (2 eq)
Cu(OAc)2 (0.1 eq)
CH2Cl2
CH2Cl2
74%
77%
Py
RT
a
Isolated yield.
1 eq.
b
2.2.2. In vivo tumor treatment studies
human hepatoma cell line (HepG2). Among these compounds,1b,1c,
1e and 1f exhibited comparable in vitro cytotoxicity with taxol, while
their invivo antitumor activity was lower than the positive control. In
addition, the cell cycle assay indicated that these novel pyridin-
2(1H)-one derivatives possessed a completely different mechanism
to CA-4, though their structures were very similar to each other.
Compounds 1b, 1c, 1e and 1f were tested in vivo against H22 cell
line. Mice were dosed once daily on 7 days of the trial with either 200
or 50 mg/kg. Tumor growth was measured and compared to tumor
growth in untreated animals and in treating with taxol (10 mg/kg). In
this model, compounds 1b, 1c and 1f (in a 50 mg/kg dose) showed
moderate tumor inhibition rate at 27%, 41%, and 31%, respectively
(Table 3). When thedosage uppedto 200 mg/kg, thetumorinhibition
rate increased accordingly. Among these tests, the death of tested
mice appeared in the group using 1b and 1c in high dose, suggesting
that 1b and 1c were more toxic than 1e and 1f. However, the in vivo
antitumor activity of the four compounds was lower than taxol.
4. Experimental section
4.1. Chemistry
1H NMR spectra were recorded on a Bruker DRX-400 (400 MHz)
or a Varian Mercury 300 NMR spectrometer. 13C NMR spectra were
obtained on a Bruker DRX-400 (100 MHz). Column chromatography
was performed on a silica gel (200e300 mesh). 4 Å Molecular sieve
was dried by microwave. The data of IR were obtained using Thermo
Electron Corporation Nicolet Nexus 670 FT-IR Spectrometer. Mass
spectra were recorded on an Agilent 1200 LC-MS (ESI) and high-
resolution mass spectra were obtained on a Waters GCT Premier
Mass Spectrometer (EI) or a Bruker Micr-OTOF-Q II spectrometer
(ESI). Melting points were taken on a FishereJohns melting point
apparatus, uncorrected and reported in degrees Centigrade.
2.2.3. Cell cycle studies
Although our designed compounds were very similar to Com-
bretastatin A-4 and its analogs, 1b, 1c, 1e and 1f did not elicit any
significant alteration to the cell cycle (Fig. 3), suggesting a distinct
mechanism of action to CA-4, which is well known to induce a G2/
M arrest of the cell cycle [26].
3. Conclusion
In summary, a series of novel 1,6-diaryl pyridin-2(1H)-one de-
rivatives were synthesized and evaluated for their antiproliferative
activities against human ovarian carcinoma cell line (SKOV-3) and
4.1.1. General procedures for the ChaneLam coupling
4.1.1.1. 6-bromo-1-(3,4,5-trimethoxyphenyl) pyridin-2(1H)-one (4).
A mixture of 6-bromopyridin-2(1H)-one (3) (8.3 g, 47.7 mmol),
(3,4,5-trimethoxyphenyl)boronic acid (20.1 g, 94.8 mmol),
Cu(OAc)2 (0.9 g, 5.0 mmol), pyridine (7.7 mL, 94.8 mmol), pyridine-
N-oxide (5.0 g, 52.6 mmol), 4 ÅMS (2.0 g) in dry CH2Cl2 (100 mL)
was stirred at room temperature overnight. Then CH2Cl2 (300 mL)
and NH.3H2O (50 mL) were added, and the reaction mixture was
filtered through Celite. The filtrate was washed with water, dried
over Na2SO4, and evaporated to dryness to yield the crude product,
which was triturated with CH2Cl2 (50 mL) to give compound 4 as a
white solid (13.0 g, 77% yield). M.p.: 174.9e176.8 ꢀC; 1H NMR
(400 MHz, CDCl3)
d
7.51 (t, J ¼ 7.8 Hz, 1H), 7.20 (d, J ¼ 7.5 Hz, 1H),
6.74 (d, J ¼ 8.1 Hz, 1H), 6.40 (s, 2H), 3.84 (s, 3H), 3.82 (s, 6H); 13C
NMR (100 MHz, CDCl3)
d 163.12, 153.79, 149.69, 141.25, 139.32,
135.28, 122.44, 109.15, 98.56, 98.50, 61.00, 56.18.
4.1.2. General procedures for the Suzuki coupling
4.1.2.1. 6-(3,4,5-trimethoxyphenyl)pyridin-2(1H)-one
mixture of compound 3 (10 g, 57.5 mmol), Pd(PPh3)4 (100 mg,
Scheme 1. Synthesis of pyridin-2(1H)-one derivatives. Reagents and conditions: (i)
3,4,5-trimethoxyphenylboronic acid, Cu(OAc)2, pyridine, pyridine-N-oxide; (ii)/(iii)
Pd(PPh3)4, K2CO3, arylboronic acid, DMF, N2 (g); (iv) arylboronic acids, Cu(OAc)2, pyr-
idine, pyridine-N-oxide.
(5). The
0.09 mmol), K2CO3 (18.2 g, 0.132 mol) and 3,4,5-

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T. Chen et al. / European Journal of Medicinal Chemistry 64 (2013) 613e620
trimethoxyphenylboronic acid (14.6 g, 68.9 mmol) in DMF (50 mL)
was refluxed under N2 (g) for 15 h. After cooled to room tempera-
ture, the mixturewas quenched with water (250 mL), extracted with
CH2Cl2 (100 ꢁ 3 mL), and evaporated to dryness to give a crude
product, which was purified by column chromatography to give 5 as
a white solid (11.5 g, 76% yield). M.p.: 194.8e196.1 ꢀC; 1H NMR
4.1.2.7. 6-(4-nitrophenyl)-1-(3,4,5-trimethoxyphenyl)pyridin-2(1H)-
one (1f). Yield: 67%. M.p.: 131.3e133.5 ꢀC. 1H NMR (400 MHz,
CDCl3)
J ¼ 7.8 Hz,1H), 7.56 (d, J ¼ 7.5 Hz,1H), 6.89 (d, J ¼ 8.2 Hz,1H), 6.49 (s,
2H), 3.88 (s, 3H), 3.84 (s, 6H); 13C NMR (100 MHz, CDCl3)
163.70,
d
8.27 (d, J ¼ 8.8 Hz, 2H), 8.11 (d, J ¼ 8.7 Hz, 2H), 7.81 (t,
d
153.80,152.93,149.90,148.24,144.20,140.49,135.36,127.46,123.94,
115.50, 110.91, 99.01, 61.04, 56.23. IR (KBr pellets) cmꢂ1: 1604.27,
1513.72, 1439.84. LC-MS (ESI) m/z ¼ 383.1 [M þ 1]þ; HRMS (EI): m/z
382.1163. Calcd for C20H18N2O6: 382.1165 [M]þ.
(400 MHz, CDCl3)
d
12.26 (br, 1H), 7.49 (m, 1H), 6.99 (s, 2H), 6.48 (m,
165.14,
2H), 3.98 (s, 6H), 3.91 (s, 3H); 13C NMR (100 MHz, CDCl3)
d
153.70, 146.95, 141.51, 139.63, 128.86, 118.17, 104.53, 103.98, 60.98,
56.34.
4.1.2.8. 6-(3-fluorophenyl)-1-(3,4,5-trimethoxyphenyl)pyridin-
2(1H)-one (1g). Yield: 74%. M.p.: 87.6e88.9 ꢀC. 1H NMR (400 MHz,
4.1.2.2. 6-(3-(benzyloxy)-4-methoxyphenyl)-1-(3,4,5-
trimethoxyphenyl)pyridin-2(1H)-one (1a). Yield: 74%. M.p.: 76.7e
CDCl3)
d
7.78e7.71 (m, 2H), 7.68 (d, J ¼ 10.4 Hz, 1H), 7.46 (d,
77.9 ꢀC. 1H NMR (400 MHz, CDCl3)
d
7.68 (t, J ¼ 7.8 Hz, 1H), 7.64 (s,
J ¼ 7.5 Hz, 1H), 7.38 (dd, J ¼ 14.1, 7.8 Hz, 1H), 7.07 (t, J ¼ 7.2 Hz, 1H),
1H), 7.55 (d, J ¼ 7.4 Hz, 1H), 7.39 (m, 5H), 7.30 (d, J ¼ 7.0 Hz,1H), 6.93
(d, J ¼ 8.4 Hz, 1H), 6.70 (d, J ¼ 8.1 Hz, 1H), 6.49 (s, 2H), 5.14 (s, 2H),
6.81 (d, J ¼ 8.2 Hz, 1H), 6.51 (s, 2H), 3.88 (s, 3H), 3.84 (s, 6H); 13C
NMR (100 MHz, CDCl3)
d 164.47, 163.39, 162.03, d 154.19 (d,
3.91 (s, 3H), 3.83 (s, 9H); 13C NMR (100 MHz, CDCl3)
d
163.31,155.15,
J ¼ 2.7 Hz), 153.70, 150.12, 140.69 (d, J ¼ 7.5 Hz), 140.28, 135.09,
130.14 (d, J ¼ 8.2 Hz), 122.23, 122.21, 115.99 (d, J ¼ 21.4 Hz), 114.74,
113.69 (d, J ¼ 23.1 Hz), 109.83, 98.91, 61.03, 56.19. IR (KBr pellets)
cmꢂ1: 1607.02, 1502.63, 1437.91. LC-MS (ESI) m/z ¼ 356.1 [M þ 1]þ;
HRMS (EI): m/z 355.1221. Calcd for C20H18FNO4: 355.1220 [M]þ.
153.65, 150.85, 150.32, 148.27, 140.07, 136.92, 134.94, 131.16, 128.51,
127.89, 127.57, 119.87, 113.82, 112.37, 111.57, 108.24, 99.01, 61.02,
56.18, 56.06. 56.05. IR (KBr pellets) cmꢂ1: 1606.25,1514.63, 1440.84.
LC-MS (ESI) m/z ¼ 474.2 [M þ 1]þ; HRMS (EI): m/z 473.1838. Calcd
for C28H27NO6: 473.1838 [M]þ.
4.1.2.9. 1-(3,4,5-trimethoxyphenyl)-[2,40-bipyridin]-6(1H)-one (1h).
4.1.2.3. 6-(3-hydroxy-4-methoxyphenyl)-1-(3,4,5-trimethoxyphenyl)
Yield: 74%. M.p.: 71.1e75.4 ꢀC. 1H NMR (400 MHz, CDCl3)
d 8.67 (d,
pyridin-2(1H)-one (1b). Yield: 74%. M.p.: 126.8e127.6 ꢀC. 1H NMR
J ¼ 5.8 Hz, 2H), 7.81 (m, 3H), 7.56 (d, J ¼ 7.5 Hz,1H), 6.90 (d, J ¼ 8.2 Hz,
(400 MHz, CDCl3)
d
7.69 (t, J ¼ 7.8 Hz, 1H), 7.53 (m, 2H), 7.39 (d,
1H), 6.49 (s, 2H), 3.88 (s, 3H), 3.84 (s, 6H); 13C NMR (100 MHz, CDCl3)
J ¼ 7.4 Hz,1H), 6.89 (d, J ¼ 8.3 Hz,1H), 6.71 (d, J ¼ 8.0 Hz,1H), 6.51 (s,
d 163.68, 153.75, 152.74, 150.44, 149.92, 145.37, 140.45, 135.22,
2H), 5.64 (s, 1H), 3.92 (s, 3H), 3.87 (s, 3H), 3.83 (s, 6H); 13C NMR
120.77, 115.16, 111.24, 98.94, 61.05, 56.21. IR (KBr pellets) cmꢂ1
:
(100 MHz, CDCl3)
d
163.22, 155.25, 153.63, 150.38, 147.65, 145.72,
1606.90,1502.35, 1440.50. LC-MS (ESI) m/z ¼ 339.1 [M þ 1]þ; HRMS
140.04, 134.90, 131.92, 118.96, 114.08, 112.91, 110.62, 108.49, 98.85,
61.03, 56.18, 56.01. IR (KBr pellets) cmꢂ1: 1599.69, 1502.58, 1439.29.
LC-MS (ESI) m/z ¼ 384.2 [M þ 1]þ; HRMS (EI): m/z 383.1366. Calcd
for C21H21NO6: 383.1369 [M]þ.
(EI): m/z 338.1265. Calcd for C19H18N2O4: 338.1267 [M]þ.
4.1.2.10. 6-(4-(trifluoromethoxy)phenyl)-1-(3,4,5-trimethoxyphenyl)
pyridin-2(1H)-one (1i). Yield: 63%. M.p.: 82.1e83.4 ꢀC. 1H NMR
(400 MHz, CDCl3)
d
7.98 (d, J ¼ 8.8 Hz, 2H), 7.75 (t, J ¼ 7.9 Hz, 1H),
4.1.2.4. 6-(4-methoxyphenyl)-1-(3,4,5-trimethoxyphenyl)pyridin-
7.46 (d, J ¼ 7.5 Hz, 1H), 7.26 (d, J ¼ 8.6 Hz, 2H), 6.80 (d, J ¼ 8.2 Hz,
2(1H)-one (1c). Yield: 85%. M.p.: 102.5e103.8 ꢀC.1H NMR (300 MHz,
1H), 6.49 (s, 2H), 3.88 (s, 3H), 3.83 (s, 6H); 13C NMR (100 MHz,
CDCl3)
d
7.92 (d, J ¼ 8.2 Hz, 2H), 7.69 (t, J ¼ 7.6 Hz, 1H), 7.41 (d,
CDCl3) d 163.50,154.17, 153.71, 150.16,149.95,140.32, 136.93, 135.07,
J ¼ 7.6 Hz,1H), 6.94 (d, J ¼ 8.4 Hz, 2H), 6.69 (d, J ¼ 8.2 Hz,1H), 6.49 (s,
128.21, 120.95, 114.56, 109.53, 98.90, 61.02, 56.17. IR (KBr pellets)
cmꢂ1: 1606.94, 1502.76, 1439.61. LC-MS (ESI) m/z ¼ 422.1 [M þ 1]þ;
HRMS (EI): m/z 421.1135. Calcd for C21H18F3NO5: 421.1137 [M]þ.
2H), 3.87 (s, 3H), 3.84 (s, 3H), 3.82 (s, 6H); 13C NMR (100 MHz, CDCl3)
d
163.32,160.63,155.38,153.63,150.39,140.06,134.89,130.96,128.06,
114.02,113.81,108.20, 98.86, 61.01, 56.16, 55.33. IR (KBr pellets) cmꢂ1
:
1606.97, 1500.95, 1439.03. LC-MS (ESI) m/z ¼ 368.2 [M þ 1]þ; HRMS
4.1.2.11. 6-(4-methoxy-3-methylphenyl)-1-(3,4,5-trimethoxyphenyl)
(EI): m/z 367.1418. Calcd for C21H21NO5: 367.1420 [M]þ.
pyridin-2(1H)-one (1j). Yield: 71%. M.p.: 85.8e87.2 ꢀC. 1H NMR
(400 MHz, CDCl3)
d
7.79 (m, 2H), 7.68 (t, J ¼ 7.8 Hz, 1H), 7.42 (d,
4.1.2.5. 6-(3,4-dimethoxyphenyl)-1-(3,4,5-trimethoxyphenyl)pyr-
J ¼ 7.6 Hz,1H), 6.86 (d, J ¼ 9.1 Hz,1H), 6.67 (d, J ¼ 8.1 Hz,1H), 6.51 (s,
idin-2(1H)-one (1d). Yield: 76%. M.p.: 118.8e120.1 ꢀC. 1H NMR
2H), 3.87 (s, 6H), 3.83 (s, 6H), 2.26 (s, 3H); 13C NMR (100 MHz,
(400 MHz, CDCl3)
d
7.70 (t, J ¼ 7.8 Hz, 1H), 7.59 (s, 1H), 7.51 (d,
CDCl3) d 163.29, 158.89, 155.72, 153.65, 150.43, 139.99, 134.76,
J ¼ 8.3 Hz,1H), 7.43 (d, J ¼ 7.5 Hz,1H), 6.90 (d, J ¼ 8.4 Hz,1H), 6.72 (d,
J ¼ 8.1 Hz,1H), 6.49 (s, 2H), 3.92 (s, 3H), 3.88 (s, 3H), 3.86 (s, 3H), 3.83
130.44, 129.10, 126.77, 125.50, 113.89, 109.88, 107.91, 98.84, 61.03,
56.17, 55.42, 16.36. IR (KBr pellets) cmꢂ1: 1603.73, 1499.01, 1428.64.
LC-MS (ESI) m/z ¼ 382.2 [M þ 1]þ; HRMS (EI): m/z 381.1574. Calcd
for C22H23NO5: 381.1576 [M]þ.
(s, 6H); 13C NMR (100 MHz, CDCl3)
d 162.37, 154.09, 152.65, 149.31,
149.17, 148.18, 139.03, 134.06, 130.28, 118.16, 112.70, 110.01, 108.86,
107.20, 98.23, 59.97, 55.18, 54.96, 54.77. IR (KBr pellets) cmꢂ1
:
1601.90, 1501.86, 1440.91. LC-MS (ESI) m/z ¼ 398.2 [M þ 1]þ; HRMS
4.1.2.12. 1,6-bis(3,4,5-trimethoxyphenyl)pyridin-2(1H)-one
Yield: 64%.1H NMR (400 MHz, CDCl3)
7.73 (t, J ¼ 7.9 Hz,1H), 7.43 (d,
J ¼ 7.6 Hz,1H), 7.21 (s, 2H), 6.76 (d, J ¼ 8.1 Hz,1H), 6.48 (s, 2H), 3.88 (s,
9H), 3.85 (s, 3H), 3.83 (s, 6H); 13C NMR (100 MHz, CDCl3)
163.29,
154.86, 153.66, 153.40, 150.23, 140.13, 139.36, 134.99, 133.71, 114.04,
(1k).
(ESI): m/z 420.1417. Calcd for C22H23NO6: 420.1423 [M þ Na]þ.
d
4.1.2.6. 6-(benzo[d][1,3]dioxol-5-yl)-1-(3,4,5-trimethoxyphenyl)pyr-
d
idin-2(1H)-one (1e). Yield: 77%. M.p.: 100.5e101.9 ꢀC. 1H NMR
(400 MHz, CDCl3)
d
7.69 (t, J ¼ 7.9 Hz,1H), 7.52e7.45 (m, 2H), 7.37 (d,
108.74,103.90, 99.27, 60.98, 60.94, 56.19, 56.11. IR (KBr pellets) cmꢂ1
:
J ¼ 7.6 Hz,1H), 6.85 (d, J ¼ 8.1 Hz,1H), 6.71 (d, J ¼ 8.1 Hz,1H), 6.49 (s,
1603.49, 1501.76, 1401.34. LC-MS (ESI) m/z ¼ 428.2 [M þ 1]þ; HRMS
2H), 5.99 (s, 2H), 3.87 (s, 3H), 3.83 (s, 6H); 13C NMR (100 MHz,
(EI): m/z 427.1628. Calcd for C23H25NO7: 427.1631 [M]þ.
CDCl3) d 163.25, 155.21, 153.66,150.31, 148.62,148.21, 140.11, 134.94,
132.84, 120.88, 114.06, 108.54, 108.36, 107.18, 101.31, 98.84, 61.03,
56.18. IR (KBr pellets) cmꢂ1: 1607.58, 1502.20, 1430.68. LC-MS (ESI)
m/z ¼ 382.1 [M þ 1]þ; HRMS (EI): m/z 381.1215. Calcd for
4.1.2.13. 6-(3-hydroxyphenyl)-1-(3,4,5-trimethoxyphenyl)pyridin-
2(1H)-one (1l). Yield: 83%. 1H NMR (400 MHz, CDCl3)
d 7.72 (t,
J ¼ 7.6 Hz, 1H), 7.46 (m, 3H), 7.26 (d, J ¼ 16 Hz, 1H), 6.87 (d,
C
21H19NO6: 381.1212 [M]þ.
J ¼ 7.5 Hz, 1H), 6.76 (d, J ¼ 8.1 Hz, 1H), 6.49 (s, 2H), 6.15 (br, 1H),

T. Chen et al. / European Journal of Medicinal Chemistry 64 (2013) 613e620
617
Table 2
Antiproliferative activity of 1aem and 2aej.
Compound
Ar
IC50
(m
M)a
Compound
Ar
IC50 (m
M)a
SKOV-3
HepG2
0.269
SKOV-3
HepG2
1a
1b
2.310
2a
2b
25.471
3.039
<0.01
<0.01
1.223
1.223
1c
<0.01
<0.01
2c
1.536
5.738
4.872
1d
0.545
0.513
2d
14.308
1e
1f
0.064
0.201
<0.01
<0.01
2e
2f
14.018
0.311
5.414
0.941
1g
0.472
0.159
2g
0.340
0.073
1h
1i
1.785
0.461
1.783
1.962
2h
2i
3.021
1.915
2.524
10.164
1j
0.583
0.844
1.461
0.129
5.459
0.034
2j
9.741
0.138
0.012
>100
1k
1m
Taxol
2.402
0.006
1l
e
a
Each value was reproduced in three experiments.
3.86 (s, 3H), 3.82 (s, 6H); 13C NMR (100 MHz, CDCl3)
d
163.34,
4.1.2.14. 6-(1H-indol-7-yl)-1-(3,4,5-trimethoxyphenyl)pyridin-
156.39, 155.31, 153.66, 150.33, 140.22, 139.81, 134.89, 129.86,
118.85, 116.40, 114.91, 113.73, 109.28, 98.88, 61.03, 56.20. IR (KBr
pellets) cmꢂ1: 1603.65, 1501.03, 1438.92. LC-MS (ESI) m/z ¼ 354.1
[M þ 1]þ; HRMS (EI): m/z 353.1264. Calcd for C20H18N2O6:
353.1263 [M]þ.
2(1H)-one (1m). Yield: 68%. M.p.: 83.7e85.1 ꢀC. 1H NMR (400 MHz,
CDCl3)
d
10.38 (s, 1H), 7.87e7.62 (m, 4H), 7.15 (t, J ¼ 7.7 Hz, 1H), 7.00
(t, J ¼ 2.6 Hz, 1H), 6.92 (d, J ¼ 7.9 Hz, 1H), 6.58 (s, 2H), 6.49 (t,
J ¼ 2.6 Hz, 1H), 3.99 (s, 3H), 3.83 (s, 6H); 13C NMR (100 MHz, CDCl3)
d
163.32, 155.55, 154.15, 150.08, 140.08, 135.49, 134.07, 129.58,

618
T. Chen et al. / European Journal of Medicinal Chemistry 64 (2013) 613e620
Table 3
Effects of 1b, 1c, 1e and 1f on mice body weight and tumor weight at pre-dose and post-dose.
Groups
Dosage (mg/kg)
No. of animals
Start
Body weight (g)
Start
Tumor weight (g)
Inhibition rate (%)
End
End
Control
1b
e
6
6
6
6
6
6
6
6
6
6
6
6
4
6
4
6
6
6
6
6
23.9 ꢃ 0.8
25.0 ꢃ 1.0
24.5 ꢃ 0.8
24.0 ꢃ 0.7
24.5 ꢃ 0.6
26.2 ꢃ 0.6
25.9 ꢃ 0.4
24.1 ꢃ 0.5
22.9 ꢃ 0.9
23.9 ꢃ 0.6
29.3 ꢃ 1.3
30.7 ꢃ 1.5
28.8 ꢃ 1.7
29.2 ꢃ 1.5
25.2 ꢃ 3.3
32.2 ꢃ 1.0
32.9 ꢃ 0.7
31.5 ꢃ 1.1
24.0 ꢃ 1.1
31.2 ꢃ 1.2
1.21 ꢃ 0.10
0.88 ꢃ 0.13
0.73 ꢃ 0.07b
0.71 ꢃ 0.14a
0.56 ꢃ 0.17a
1.01 ꢃ 0.16
0.89 ꢃ 0.15
0.84 ꢃ 0.12a
0.47 ꢃ 0.03c
0.64 ꢃ 0.15a
e
50
200
50
200
50
200
50
27
40
41
54
17
27
31
61
47
1c
1e
1f
Taxol
200
10
a
p < 0.05.
p < 0.01.
p < 0.001.
b
c
124.59, 122.65, 119.99, 119.25, 119.20, 113.56, 108.03, 101.76, 100.17,
61.14, 56.23. IR (KBr pellets) cmꢂ1: 1603.60,1499.90,1441.43. LC-MS
(ESI) m/z ¼ 377.2 [M þ 1]þ; HRMS (EI): m/z 376.1426. Calcd for
4.1.2.19. 1-(benzo[d][1,3]dioxol-5-yl)-6-(3,4,5-trimethoxyphenyl)pyr-
idin-2(1H)-one (2e). Yield: 33%. M.p.: 113.3e114.9 ꢀC. 1H NMR
(400 MHz, CDCl3)
7.20 (s, 2H), 6.81 (m, 2H), 6.74 (d, J ¼ 8.1 Hz, 1H), 6.68 (dd, J ¼ 8.3,
d
7.70 (t, J ¼ 7.8 Hz, 1H), 7.40 (d, J ¼ 7.5 Hz, 1H),
C
22H20N2O4: 376.1423 [M]þ.
2.3 Hz, 1H), 5.98 (s, 2H), 3.89 (s, 6H), 3.87 (s, 3H); 13C NMR
4.1. 2.15. 1-(3-(benzyloxy)-4-methoxyphenyl)-6-(3,4,5-
(100 MHz, CDCl3) d 163.54, 154.77, 153.35, 148.50, 147.99, 144.46,
trimethoxyphenyl)pyridin-2(1H)-one (2a). Yield: 42%. M.p.: 86.1e
140.04, 139.17, 133.88, 114.13, 113.90, 108.64, 107.98, 104.23, 103.92,
101.54, 60.93, 56.07. IR (KBr pellets) cmꢂ1: 1579.10, 1485.12,
1402.13. LC-MS (ESI) m/z ¼ 382.1 [M þ 1]þ; HRMS (EI): m/z
381.1209. Calcd for C21H19NO6: 381.1212 [M]þ.
87.6 ꢀC. 1H NMR (300 MHz, CDCl3)
d
7.66 (t, J ¼ 7.9 Hz, 1H), 7.43e
7.27 (m, 6H), 7.19 (s, 2H), 6.91 (d, J ¼ 8.6 Hz, 1H), 6.80 (m, 2H),
6.63 (d, J ¼ 8.1 Hz, 1H), 5.11 (s, 2H), 3.89 (s, 3H), 3.87 (s, 9H); 13C
NMR (100 MHz, CDCl3)
d 163.73, 154.89, 153.36, 148.77, 147.70,
146.91, 140.02, 139.21, 136.63, 133.89, 128.63, 128.56, 128.09,
127.95, 127.42, 127.40, 124.01, 117.32, 113.86, 113.72, 113.14, 112.20,
108.77, 108.38, 103.96, 71.12, 60.93, 56.51, 56.13. IR (KBr pellets)
4.1.2.20. 1-(4-nitrophenyl)-6-(3,4,5-trimethoxyphenyl)pyridin-
2(1H)-one (2f). Yield: 32%. M.p.: 133.8e134.3 ꢀC. 1H NMR
(400 MHz, CDCl3)
d
8.29 (d, J ¼ 8.9 Hz, 2H), 7.83 (t, J ¼ 7.8 Hz, 1H),
cmꢂ1
:
1584.61, 1502.56, 1440.85. LC-MS (ESI) m/z
¼
474.2
7.53 (d, J ¼ 7.5 Hz, 1H), 7.38 (d, J ¼ 8.9 Hz, 2H), 7.13 (s, 2H), 6.93 (d,
[M þ 1]þ; HRMS (EI): m/z 473.1835. Calcd for C28H27NO6:
J ¼ 8.0 Hz, 1H), 3.87 (s, 3H), 3.86 (s, 6H); 13C NMR (100 MHz, CDCl3)
473.1838 [M]þ.
d 161.78, 159.68, 155.02, 153.42, 143.78, 140.72, 133.10, 125.19,
121.18, 116.19, 115.55, 110.33, 103.95, 60.95, 56.13. IR (KBr pellets)
cmꢂ1: 1594.50, 1401.63. LC-MS (ESI) m/z ¼ 383.1 [M þ 1]þ; HRMS
(EI): m/z 382.1168. Calcd for C20H18N2O6: 382.1165 [M]þ.
4.1.2.16. 1-(3-hydroxy-4-methoxyphenyl)-6-(3,4,5-trimethoxyphenyl)
pyridin-2(1H)-one (2b). Yield: 78%. 1H NMR (400 MHz, CDCl3)
d 7.68
(t, J ¼ 7.9 Hz,1H), 7.38 (d, J ¼ 7.5 Hz,1H), 7.21 (s, 2H), 6.90 (d, J ¼ 2.7 Hz,
1H), 6.85 (d, J ¼ 8.7 Hz, 1H), 6.77e6.68 (m, 2H), 3.90 (s, 3H), 3.89 (s,
4.1.2.21. 1-(3-fluorophenyl)-6-(3,4,5-trimethoxyphenyl)pyridin-
2(1H)-one (2g). Yield: 44%. M.p.: 62.7e64.4 ꢀC. 1H NMR (400 MHz,
6H), 3.87 (s, 3H); 13C NMR (100 MHz, CDCl3)
d 163.58,154.88,153.33,
148.15, 146.19, 143.78, 139.97, 139.13, 134.01, 113.89, 112.54, 110.82,
CDCl3)
d
7.75 (t, J ¼ 7.8 Hz, 1H), 7.44 (d, J ¼ 7.5 Hz, 1H), 7.34 (dd,
109.07, 108.61, 104.00, 60.92, 56.32, 56.10. IR (KBr pellets) cmꢂ1
:
J ¼ 14.8, 8.2 Hz, 1H), 7.17 (s, 2H), 7.04 (t, J ¼ 10.1 Hz, 2H), 6.91 (t,
1585.35, 1502.84, 1401.28. LC-MS (ESI) m/z ¼ 384.2 [M þ 1]þ; HRMS
J ¼ 8.3 Hz, 1H), 6.83 (d, J ¼ 8.1 Hz, 1H), 3.87 (s, 9H); 13C NMR
(EI): m/z 383.1363. Calcd for C21H21NO6: 383.1369 [M]þ.
(100 MHz, CDCl3)
d 162.52, 154.70, 153.39, 140.24, 139.34, 133.62,
129.96 (d, J ¼ 9.8 Hz), 117.23, 114.37, 111.35, 111.14, 109.63, 109.46,
109.39, 103.92, 60.90, 56.06. IR (KBr pellets) cmꢂ1: 1573.11, 1503.20,
1430.28. LC-MS (ESI) m/z ¼ 356.1 [M þ 1]þ; HRMS (EI): m/z
355.1221. Calcd for C20H18FNO4: 355.1220 [M]þ.
4.1.2.17. 1-(4-methoxyphenyl)-6-(3,4,5-trimethoxyphenyl)pyridin-
2(1H)-one (2c). Yield: 35%. M.p.: 75.4e76.8 ꢀC. 1H NMR (300 MHz,
CDCl3)
4H), 6.93 (d, J ¼ 9.1 Hz, 2H), 6.72 (d, J ¼ 8.1 Hz, 1H), 3.87 (s, 9H), 3.81
(s, 3H); 13C NMR (100 MHz, CDCl3)
163.77, 156.59, 154.65, 153.33,
d
7.69 (t, J ¼ 8.5 Hz, 1H), 7.39 (d, J ¼ 7.5 Hz, 1H), 7.20e7.10 (m,
d
4.1.2.22. 6-(3,4,5-trimethoxyphenyl)-2H-[1,40-bipyridin]-2-one (2h).
147.58, 139.93, 139.22, 133.88, 122.76, 114.40, 113.60, 108.55, 103.96,
60.89, 56.10, 55.64. IR (KBr pellets) cmꢂ1: 1584.07, 1503.10, 1439.27.
LC-MS (ESI) m/z ¼ 368.2 [M þ 1]þ; HRMS (EI): m/z 367.1417. Calcd
for C21H21NO5: 367.1420 [M]þ.
Yield: 33%. M.p.: 187.9e189.5 ꢀC. 1H NMR (400 MHz, CDCl3)
d 8.58
(d, J ¼ 4.9 Hz, 2H), 7.81 (t, J ¼ 7.8 Hz, 1H), 7.53 (d, J ¼ 7.6 Hz, 1H), 7.18
(d, J ¼ 5.2 Hz, 2H), 7.16 (s, 2H), 6.91 (d, J ¼ 8.0 Hz, 1H), 3.87 (s, 9H);
13C NMR (100 MHz, CDCl3)
d 161.42, 161.05, 155.05, 153.45, 151.19,
140.68, 139.43, 133.35, 115.72, 115.61, 110.65, 103.86, 60.94, 56.12. IR
(KBr pellets) cmꢂ1: 1642.78, 1605.23, 1401.61. LC-MS (ESI) m/
z ¼ 339.1 [M þ 1]þ; HRMS (EI): m/z 338.1267. Calcd for C19H18N2O4:
338.1266 [M]þ.
4.1.2.18. 1-(3,4-dimethoxyphenyl)-6-(3,4,5-trimethoxyphenyl)pyr-
idin-2(1H)-one (2d). Yield: 26%. M.p.: 84.1e85.7 ꢀC. 1H NMR
(400 MHz, CDCl3)
d
7.70 (t, J ¼ 7.7 Hz, 1H), 7.41 (d, J ¼ 7.5 Hz, 1H),
7.20 (s, 2H), 6.89 (d, J ¼ 8.5 Hz, 1H), 6.83e6.76 (m, 2H), 6.73 (d,
J ¼ 8.1 Hz, 1H), 3.90 (s, 3H), 3.88 (s, 9H), 3.86 (s, 3H); 13C NMR
4.1.2.23. 1-(4-(trifluoromethyl)phenyl)-6-(3,4,5-trimethoxyphenyl)
pyridin-2(1H)-one (2i). Yield: 41%. M.p.: 111.2e112.6 ꢀC. 1H NMR
(100 MHz, CDCl3)
d 163.77, 154.85, 153.35, 149.61, 147.79, 146.17,
140.04, 139.17, 133.89, 113.84, 113.08, 111.34, 108.46, 106.21, 103.88,
60.94, 56.29, 56.10, 56.03. IR (KBr pellets) cmꢂ1: 1599.75, 1512.19,
1447.41. LC-MS (ESI) m/z ¼ 398.2 [M þ 1]þ; HRMS (ESI): m/z
420.1416. Calcd for C22H23NO6: 420.1423 [M þ Na]þ.
(400 MHz, CDCl3)
d
7.77 (t, J ¼ 7.8 Hz, 1H), 7.67 (d, J ¼ 8.5 Hz, 2H),
7.47 (d, J ¼ 7.6 Hz, 1H), 7.34 (d, J ¼ 8.4 Hz, 2H), 7.11 (s, 2H), 6.88 (d,
J ¼ 8.1 Hz, 1H), 3.86 (s, 3H), 3.82 (s, 6H); 13C NMR (100 MHz,
CDCl3)
d 162.44, 156.98, 154.51, 153.38, 140.42, 139.33, 133.38,

T. Chen et al. / European Journal of Medicinal Chemistry 64 (2013) 613e620
619
Fig. 3. Cell cycle analysis of HepG2 cells.
126.57, 126.53, 122.04, 114.43, 109.59, 103.70, 60.92, 55.98. IR (KBr
pellets) cmꢂ1: 1572.57, 1510.35, 1440.86. LC-MS (ESI) m/z ¼ 406.1
[M þ 1]þ; HRMS (EI): m/z 405.1190. Calcd for C21H18F3NO4:
405.1188 [M]þ.
4.1.2.24. 1-(4-fluorophenyl)-6-(3,4,5-trimethoxyphenyl)pyridin-
2(1H)-one (2j). Yield: 38%. M.p.: 79.8e81.4 ꢀC. 1H NMR (400 MHz,
CDCl3)
d
7.73 (t, J ¼ 7.8 Hz, 1H), 7.42 (d, J ¼ 7.5 Hz, 1H), 7.20 (m, 2H),
7.14 (s, 2H), 7.09 (m, 2H), 6.79 (d, J ¼ 8.1 Hz, 1H), 3.87 (s, 3H), 3.86 (s,

620
T. Chen et al. / European Journal of Medicinal Chemistry 64 (2013) 613e620
6H); 13C NMR (100 MHz, CDCl3)
d
163.21, 160.81, 158.39, 154.52,
grants of The Fundamental Research Funds for the Central Uni-
153.34, 149.90, 140.16, 139.19, 133.68, 123.32, 123.23, 115.92, 115.69,
113.93, 108.94, 103.76, 60.93, 56.05. IR (KBr pellets) cmꢂ1: 1589.02,
1501.64, 1442.61. LC-MS (ESI) m/z ¼ 356.1 [M þ 1]þ; HRMS (EI): m/z
355.1222. Calcd for C20H18FNO4: 355.1220 [M]þ.
versities. We also thank State key Laboratory of Drug Research,
shanghai Institute of Materia, Chinese Academy of Sciences for
support.
Appendix A. Supplementary data
4.2. Antiproliferative assays
Supplementary material associated with this article can be
found, in the online version, at http://dx.doi.org/10.1016/j.
ejmech.2013.04.008.
Cells lines (human ovarian carcinoma cell line SKOV-3 and hu-
man hepatoma cell line HepG2) were maintained in the same
standard medium and grown as a monolayer in DMEM (Gibco,
Grand Island, NY, USA) and supplemented with 10% fetal bovine
serum (FBS) and antibiotics, i.e.,100 IU/mL penicillin and 0.1 mg/mL
streptomycin. Culture was maintained at 37 ꢀC with 5% CO2 in a
humidified atmosphere. Cells (5 ꢁ 103 cells/well) were treated as
indicated for 72 h in medium containing 10% FBSC. The medium
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another day. Fifty microliter of serum-free medium containing XTT
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m
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This work was supported by the grants of The National Natural
Science Foundation of China (Nos. 81172936 and 21102046) and