Synthesis and biological evaluation of some novel resveratrol amide derivatives as potential anti-tumor agents

Article abstract of DOI:10.1016/j.ejmech.2012.11.021

Three series of novel resveratrol amide derivatives (1a-q, 2a-h, 3a-l) were synthesized and evaluated for their biological activities. All compounds were characterized by ¹H NMR, ¹³C NMR, MS and elemental analysis. Furthermore, compo

Full text of DOI:10.1016/j.ejmech.2012.11.021

European Journal of Medicinal Chemistry 62 (2013) 222e231
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 some novel resveratrol amide
derivatives as potential anti-tumor agents
,
Ri-Sheng Yaoa, Xiao-Qin Lu a, Qiu-Xiang Guana, Lei Zheng b, Xiang Luc, Ban-Feng Ruan a
*
a School of Medical Engineering, Hefei University of Technology, Hefei, PR China
b School of Biology and Food Engineering, Hefei University of Technology, Hefei, PR China
c State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, PR China
a r t i c l e i n f o
a b s t r a c t
Article history:
Three series of novel resveratrol amide derivatives (1aeq, 2aeh, 3ael) were synthesized and evaluated
for their biological activities. All compounds were characterized by 1H NMR, 13C NMR, MS and elemental
analysis. Furthermore, compound 3e was also characterized by X-ray crystallography. All the compounds
were evaluated for their anti-tumor activity against MCF-7, A549 and B16-F10 tumor cell lines as well as
cyclooxygenase-2 (COX-2)-derived prostaglandin E2 (PGE2) inhibitory activity of murine macrophage
RAW 264.7 cell line. Among them, compounds 1c, 1g and 3e displayed the most potent COX-2 inhibitory
Received 31 May 2012
Received in revised form
19 October 2012
Accepted 14 November 2012
Available online 20 November 2012
activity with the IC50 values of 1.02, 1.27 and 1.98
performed to position compounds 1c and 3e into the active site of COX-2 to determine the probable
binding modes.
mM, respectively. Molecular docking studies were
Keywords:
Resveratrol derivatives
COX-2 inhibitor
Anti-tumor
Ó 2012 Elsevier Masson SAS. All rights reserved.
Molecular docking
1. Introduction
Several selective COX-2 inhibitors are currently used in the
clinics (e.g. celecoxib, valdecoxib, parecoxib, rofecoxib, etoricoxib,
Cyclooxygenase (COX) enzymes have the key role in catalyzing
biotransformation of arachidonic acid (AA) to prostaglandins (PGs),
prostacyclin (PGI2), and thromboxane A2 (TXA2) [1]. COX activity
originates from two distinct and independently regulated enzymes,
termed COX-1 and COX-2. COX-1 is the constitutive isoform and is
mainly responsible for the synthesis of cytoprotective prostaglan-
dins in the gastrointestinal (GI) tract and thromboxane which
triggers platelet aggregation in blood platelets. COX-2 is inducible
and short-lived; its expression is stimulated in response to endo-
toxins, cytokines, and mitogens [2].
The role of COX-2 is very important. COX-2 has been implicated
in human carcinogenesis, and thus, inhibition of COX-2 might have
the dual effect on controlling both inflammation and cancer [3].
Numerous studies have demonstrated the overexpression of COX-2
in solid malignancies [4]. Epidemiological, clinical, and preclinical
investigations also provide compelling evidence that COX-2
inhibitors could act as chemopreventive agents [5]. Recent data
have demonstrated the involvement of COX-2 in both in vitro
proliferation and in vivo tumor growth rate [6,7]. Other works have
highlighted the role played by COX-2 in disturbing the balance
between matrix metalloproteinases (MMPs) and the tissue inhibi-
tors of metalloproteinases (TIMPs) in prostate cancer cells [8,9].
and lumiracoxib) which provide effective treatment of inflamma-
tory disease states such as rheumatoid arthritis and osteoarthritis
[10]. Several lines of evidence suggest that these selective COX-2
inhibitors may also provide an opportunity for both cancer
prevention and therapy [11].
Natural products in general and medicinal plants in particular,
are believed to be an important source of new chemical substances
with a potential therapeutic efficacy. Thus many researchers
have investigated selective COX-2 inhibitors in natural products.
Among them, resveratrol (3,4,50-trihydroxy-trans-stilibene) is
a phytoalexin found mainly in the skin of grapes and red wine and
exhibits anticancer, antioxidation, anti-inflammatory, cardiovas-
cular protection properties. It has also been shown to be a non-
selective inhibitor of COX-1 and COX-2 [12,13]. In order to find
more selective COX-2 inhibitors, three series of resveratrol deriva-
tives (1aeq, 2aeh and 3ael) were synthesized and evaluated for
their anticancer, anti-inflammatory and COX-2 inhibition activities.
In order to identify the possible binding mode, molecular docking
studies were consequently performed.
2. Chemistry
Three series of compounds were synthesized via the route
outlined in Scheme 1. The intermediate (E)-2,4-dimethoxy-6-(4-
* Corresponding author. Tel.: þ86 551 2901771.
E-mail address: ruanbf@hfut.edu.cn (B.-F. Ruan).
0223-5234/$ e see front matter Ó 2012 Elsevier Masson SAS. All rights reserved.
http://dx.doi.org/10.1016/j.ejmech.2012.11.021

R.-S. Yao et al. / European Journal of Medicinal Chemistry 62 (2013) 222e231
223
O
O
H
N
R1
O
O
q
1a-
O
O
O
O
O
O
R2
N
H
C
O
i
R3
ii
H
COO
O
O
A
O
O
B
2a-h
O
O
R4
O
O
3a-l
Scheme 1. Reagents and conditions: (i) malonate, pyridine, piperidine, 95 ꢀC; (ii) EDCI, HOBt, DCM, appropriate substituted amines, room temperature.
methoxystyryl) benzaldehyde (A) was obtained as reported [14].
(E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)acrylic acid
(B) was obtained in 92.1% yield by Knoevenagel condensation. The
target compounds (1aeq, 2aeh and 3ael) were prepared by
reaction of B with primary, secondary and cyclic amines in good
yields, respectively.
All the obtained compounds gave satisfactory elementary
analytical and spectroscopic data. 1H NMR, 13C NMR and ESI MS
spectra were consistent with the assigned structures (Table 1).
Furthermore, compound 3e was also determined by single crystal
X-ray diffraction analysis.
as well as the addition of branched groups. Compound 1g has the
comparable activity to 1c, probably because of the similar length of
3-chloropropal to butyl. So the length of the side chain (R1) played
a key role in the activities. Compounds 2aeh showed almost the
same moderate anticancer activities, and they were not as good as
1c. This demonstrated that the substitution of secondary amines
had little effect on the activities. The difference among compounds
3aee lies in the position and number of methyl on piperidinyl.
Among them, 3e (R4 ¼ 3, 5-dimethylpiperidinyl) showed the potent
activity with IC50 value of 1.57, 2.27 and 3.26
mM when tested
against MCF-7, A549 and B16-F10, respectively. And they follow the
sequence of 3,5-dimethyl > ortho-methyl > para-methyl > meta-
methyl. The change of methyl to other groups (3feh) or piperidine
to other cyclic amines (3iel) could not increase the activities. These
changes of R4 were associated with great differences in activity,
which suggested a possible critical binding role for this group in the
binding site of the molecular target [15].
To study the effect of the synthesized compounds on COX-2
selectivity and potency, we determined the ability of compounds
to inhibit the COX-1 and COX-2 isozymes using chemiluminescent
enzyme assays kit. The efficacies of compounds were determined as
the concentration causing 50% enzyme inhibition (IC50). The results
were summarized in Table 2. These data showed that most of the
synthesized compounds exhibited remarkable and selective inhi-
bition of COX-2 isozyme, while weak to COX-1, and particularly,
compound 1c, 1g, 3e and 3j showed significant activities compa-
rable to celecoxib. Among them, 1c was the most potent
3. Results and discussion
3.1. Biological activity
It is well known that COX-2 is overexpressed in many human
cancer entities such as breast cancer, hepatic carcinoma and
melanoma. Thirty-seven target compounds were evaluated for the
antiproliferative activities against human breast cancer MCF-7
cells, human non-small cell lung cancer A549 cells and melanoma
B16-F10 cells. The results were presented in Table 2. All compounds
showed obvious activities with IC50 values between 1.33 and
13.63
mM, and they were much better than that of the positive
control celecoxib. Compounds 1c, 1g and 3e had significant
activities.
Subsequently SAR studies were performed to determine how
the substituents (R1eR4) affected the antiproliferative activities.
Among the first series of compounds 1aeq, 1c (R1 ¼ butyl) dis-
(IC50 ¼ 1.02
western blotting assay shown in Fig. 1 confirmed that compound 1c
with the concentration above 0.5 M could obviously inhibit lipo-
mM) COX-2 inhibitor. Furthermore, the results of
played the most potent activity (IC50 ¼ 1.33
m
M for MCF-7; 1.88
m
M
m
for A549; 2.08
m
M for B16-F10). It can be seen that the activities
polysaccharide (LPS)-induced COX-2 expression in murine macro-
phage RAW 264.7 cell line.
would be reduced by the increase or decrease of the length of butyl

224
R.-S. Yao et al. / European Journal of Medicinal Chemistry 62 (2013) 222e231
Prostaglandin E2 (PGE2) is biosynthesized via COX-2 enzymes,
which is one of the principal mediators of inflammation.
Compound 1c exhibited promising inhibitory against PGE2 with
Table 1
Chemical structures of the synthetic compounds.
Structure
Comp. R1
no
IC50 value of 0.28
mM as shown in Table 2, which was approximate
to that of celecoxib (IC50 ¼ 0.10
m
M). These data suggested that
these synthesized compounds inhibited the inflammatory activity
via the COX-2 pathway and were devoid of toxicity due to the
absence of COX-1 inhibition which maintains normal physiological
functions.
1a
1b
1c
1d
1e
1f
3.2. Crystal structure of compound 3e
1g
1h
1i
1j
1k
Compound 3e crystallizes in the monoclinic space group P2(1)/
n. The crystal data and refinement data are listed in Table 3.
Selected bond lengths and angles are given in Table 4. All bond
lengths are within normal ranges [16]. As shown in Fig. 2a, C10eC9
and C19eC20 are both in trans form and the bond lengths of
1l
1m
ꢀ
1n
1.332(4) and 1.326(5) A conform to the value for double CeC bond,
respectively. Similarly, the C8eO1 bond length of 1.225(5)
conforms to the value for double CeO bond. The dihedral angle
between the two phenyl rings is 56.7(1)ꢀ (C21eC22eC23eC24e
C25eC26 and C11eC12eC13eC14eC15eC16). The six-membered
piperidine ring is in a standard chair form. As shown in Fig. 2b,
intermolecular H bonds (H27B/O1) between adjacent molecules
lead to a 1D chain structure. Furthermore, intermolecular H bonds
(H26/O1 and H23/O2) between adjacent 1D chains generate
a 2D layer.
1o
1p
1q
R2
R3
2a
2b
3.3. Molecular docking
2c
2d
In order to get a better insight into the binding affinity and guide
further SAR studies, molecular docking studies of compounds 1c and
3e were performed using the crystal structure of COX-2 enzymes
given in PDB code 1cx2. All docking runs were applied the
Lamarckian genetic algorithm of Auto-Dock 4.0. The binding modes
of 1c and 3e were depicted in Fig. 3. It can be seen that the C]O of
compound 1c, formed a hydrogen bonding interaction with the
2e
2f
2g
2h
R4
3a
3b
ꢀ
backbone NH of Arg120 (bond length: Arg120 NeH/O ¼ 2.037A;
bond angle: Arg120 NeH/O ¼ 160.073ꢀ). Compound 3e as well as
ꢀ
1c, interacts with Arg120 (bond length: Arg120 NeH/O ¼ 1.968A;
bond angle: Arg120 NeH/O ¼ 152.713ꢀ), and 3e had one more
interaction. The oxygen atom of one of the methoxyl groups on
benzene ring formed a hydrogen bond with Lys83 (bond lengtꢀh:
3c
ꢀ
3d
Lys83 NeH/O ¼ 1.637A; bond angle: Lys83 NeH/O ¼ 131.135 ).
The 3D binding modes of 1c and 3e were exhibited in Fig. 4. They
also showed well binding affinity to the target via hydrogen bonds
and hydrophobic interactions.
3e
3f
These results of molecular docking studies proved that
compounds 1c and 3e had high binding potency of COX-2.
4. Conclusion
3g
Three series of amide derivatives of resveratrol were synthe-
sized and evaluated for the anti-tumor and COX-1/COX-2 inhibition
activity. Compounds 1c, 1g and 3e exhibited the most potent anti-
tumor activity against MCF-7, A549 and B16-F10 tumor cell lines.
Furthermore, compounds 1c, 1g and 3e also exhibited optimal COX-
2 inhibitory potency and selectivity compared with celecoxib.
Molecular docking studies further help in understanding the
interactions between the ligands and enzyme active sites in detail
and thereby help to design novel potent inhibitors from natural
product. It is clear that compounds 1c and 3e interact with the
binding site of COX-2 by forming strong hydrogen bond(s) and
hydrophobic interactions. The results of this work might be helpful
for discovering novel class of potent anti-tumor agents.
3h
3i
3j
3k
3l

R.-S. Yao et al. / European Journal of Medicinal Chemistry 62 (2013) 222e231
225
Table 2
Inhibition (IC50) of MCF-7, A549 and B16-F10 tumor cell lines as well as the inhibition of COX-1, PGE2 and COX-2 in lipopolysaccharide (LPS)-activated murine macrophage RAW
264.7 cells.
Comp. no
IC50
(
m
M)
MCF-7
A549
B16-F10
COX-1
COX-2
PGE2
7.06 ꢁ 1.50
1a
1b
1c
1d
1e
1f
1g
1h
1i
7.06 ꢁ 0.21
5.61 ꢁ 0.15
1.33 ꢁ 0.06
5.53 ꢁ 0.13
4.28 ꢁ 0.08
2.30 ꢁ 0.14
1.98 ꢁ 0.08
4.56 ꢁ 0.96
2.56 ꢁ 1.25
13.63 ꢁ 0.06
5.29 ꢁ 0.36
3.43 ꢁ 0.73
3.94 ꢁ 0.02
2.63 ꢁ 0.11
5.42 ꢁ 0.15
4.15 ꢁ 0.08
7.01 ꢁ 0.03
6.86 ꢁ 0.16
6.55 ꢁ 0.01
2.00 ꢁ 0.22
3.99 ꢁ 0.03
2.39 ꢁ 0.05
6.13 ꢁ 0.23
4.03 ꢁ 0.02
3.22 ꢁ 0.11
6.05 ꢁ 0.11
4.35 ꢁ 0.13
11.92 ꢁ 0.12
6.45 ꢁ 0.38
1.57 ꢁ 0.10
5.24 ꢁ 0.21
4.95 ꢁ 0.11
4.23 ꢁ 0.07
2.29 ꢁ 0.06
3.31 ꢁ 0.70
2.76 ꢁ 0.06
4.2 ꢁ 0.34
7.86 ꢁ 0.81
5.17 ꢁ 0.34
1.88 ꢁ 0.31
6.23 ꢁ 0.48
8.66 ꢁ 0.56
2.00 ꢁ 0.23
1.98 ꢁ 0.11
6.93 ꢁ 1.13
2.36 ꢁ 0.25
8.67 ꢁ 0.42
10.44 ꢁ 1.14
3.95 ꢁ 0.28
8.81 ꢁ 0.68
3.61 ꢁ 0.62
6.42 ꢁ 0.46
4.22 ꢁ 1.00
5.14 ꢁ 10.6
6.00 ꢁ 0.76
3.35 ꢁ 0.49
5.58 ꢁ 0.15
6.16 ꢁ 0.38
3.35 ꢁ 0.51
7.49 ꢁ 0.84
5.34 ꢁ 0.61
5.89 ꢁ 0.63
10.62 ꢁ 1.28
5.77 ꢁ 0.74
5.82 ꢁ 0.33
7.20 ꢁ 0.88
2.27 ꢁ 0.14
5.60 ꢁ 0.16
2.08 ꢁ 0.38
6.69 ꢁ 0.45
4.51 ꢁ 0.14
2.32 ꢁ 0.28
5.40 ꢁ 0.93
6.35 ꢁ 0.98
5.46 ꢁ 0.39
15.6 ꢁ 1.38
6.99 ꢁ 0.44
9.08 ꢁ 1.06
2.08 ꢁ 0.27
7.18 ꢁ 0.62
8.06 ꢁ 1.07
3.45 ꢁ 0.29
2.78 ꢁ 0.31
8.28 ꢁ 0.94
4.02 ꢁ 0.42
7.98 ꢁ 0.87
12.65 ꢁ 1.24
4.38 ꢁ 0.54
10.16 ꢁ 1.48
5.48 ꢁ 0.69
10.09 ꢁ 1.16
6.83 ꢁ 0.76
8.15 ꢁ 0.81
5.78 ꢁ 0.52
3.98 ꢁ 0.23
5.98 ꢁ 0.47
9.03 ꢁ 0.96
4.85 ꢁ 0.12
9.27 ꢁ 0.82
8.02 ꢁ 0.94
8.18 ꢁ 0.78
9.75 ꢁ 1.04
7.66 ꢁ 1.24
8.25 ꢁ 0.79
7.76 ꢁ 1.36
3.26 ꢁ 0.27
7.38 ꢁ 0.69
4.69 ꢁ 0.64
9.18 ꢁ 1.52
5.67 ꢁ 0.91
3.08 ꢁ 0.28
8.10 ꢁ 1.04
9.28 ꢁ 0.93
6.78 ꢁ 0.64
85.9 ꢁ 2.34
35.36 ꢁ 1.86
>50
>50
12.36 ꢁ 1.50
6.75 ꢁ 0.26
1.02 ꢁ 0.30
8.99 ꢁ 0.91
20.09 ꢁ 2.85
1.36 ꢁ 0.43
1.27 ꢁ 0.08
9.08 ꢁ 1.17
2.01 ꢁ 0.49
11.12 ꢁ 1.84
15.09 ꢁ 2.04
4.56 ꢁ 0.46
12.96 ꢁ 2.74
5.09 ꢁ 1.12
9.67 ꢁ 1.36
8.92 ꢁ 1.03
6.66 ꢁ 0.98
12.98 ꢁ 1.88
4.67 ꢁ 0.45
8.06 ꢁ 1.58
10.08 ꢁ 2.04
6.84 ꢁ 1.04
16.43 ꢁ 1.78
9.08 ꢁ 0.91
8.01 ꢁ 1.47
23.16 ꢁ 2.68
6.69 ꢁ 0.74
7.09 ꢁ 1.28
10.28 ꢁ 2.07
1.98 ꢁ 0.09
10.28 ꢁ 2.08
1.45 ꢁ 0.11
8.57 ꢁ 1.58
5.78 ꢁ 0.44
1.88 ꢁ 0.14
8.01 ꢁ 1.06
8.87 ꢁ 1.28
6.89 ꢁ 0.91
0.10 ꢁ 0.02
2.08 ꢁ 0.12
0.28 ꢁ 0.04
5.01 ꢁ 0.82
13.82 ꢁ 1.95
0.45 ꢁ 0.07
0.39 ꢁ 0.14
5.08 ꢁ 0.67
0.72 ꢁ 0.06
5.29 ꢁ 0.89
6.12 ꢁ 0.64
2.09 ꢁ 0.26
7.05 ꢁ 1.15
1.98 ꢁ 0.38
3.15 ꢁ 0.57
3.68 ꢁ 0.48
2.10 ꢁ 0.07
4.36 ꢁ 0.26
2.15 ꢁ 0.31
3.88 ꢁ 0.10
4.12 ꢁ 0.29
2.56 ꢁ 0.72
7.19 ꢁ 1.06
3.10 ꢁ 0.83
3.28 ꢁ 0.54
15.29 ꢁ 2.81
2.44 ꢁ 0.43
3.16 ꢁ 0.46
8.09 ꢁ 1.04
0.58 ꢁ 0.03
6.27 ꢁ 0.48
0.46 ꢁ 0.08
3.55 ꢁ 0.46
1.62 ꢁ 0.32
0.85 ꢁ 0.06
3.18 ꢁ 0.40
3.76 ꢁ 0.47
3.01 ꢁ 0.29
0.12 ꢁ 0.01
34.68 ꢁ 2.48
>50
32.26 ꢁ 2.64
38.39 ꢁ 3.28
>50
40.39 ꢁ 2.89
1j
1k
1l
>50
>50
>50
>50
>50
1m
1n
1o
1p
1q
2a
2b
2c
2d
2e
2f
2g
2h
3a
3b
3c
3d
3e
3f
3g
3h
3i
3j
3k
3l
29.09 ꢁ 3.58
>50
>50
>50
>50
45.88 ꢁ 2.96
>50
>50
>50
>50
34.98 ꢁ 2.74
>50
>50
>50
>50
45.06 ꢁ 3.58
>50
40.56 ꢁ 4.26
>50
39.06 ꢁ 2.81
>50
45.19 ꢁ 4.68
>50
>50
27.5 ꢁ 2.46
B
6.10 ꢁ 0.36
40.8 ꢁ 0.42
Celecoxib
5. Experimental
5.1. General
Table 3
Crystallographic data and structure refinements for compound 3e.
All the chemicals used were commercial products employed
without further purification. The 1H NMR spectra were recorded
on a Bruker DRX 300 model spectrometer in CDCl3 solutions at
room temperature with TMS as an internal standard. The 13C NMR
spectra were recorded on a Bruker DRX 600 model spectrometer in
CDCl3 solutions at room temperature with TMS as an internal
Compound
3e
Formula
C
27H33NO4
Formula wt
Crystal system
Space group
Crystal size (mm3)
435.54
monoclinic
P2(1)/n
0.30 ꢂ 0.20 ꢂ 0.20
13.273(5)
8.619(5)
22.032(5)
90.000(5)
99.450(5)
90.000(5)
2486.3(18)
4
standard. Chemical shifts (d
) for 1H NMR and 13C NMR spectra were
reported in parts per million to residual solvent protons. Melting
points were measured on a Boetius micro melting point apparatus.
ꢀ
a (A)
ꢀ
b (A)
ꢀ
c (A)
a
b
g
(ꢀ)
(ꢀ)
(ꢀ)
3
ꢀ
V (A )
Z
Dc (g cmꢃ3
)
1.164
0.077
m
(mmꢃ1
)
F (000)
q
936
2.54e21.20
16,842
range (ꢀ)
Reflns collected
Reflns unique
4381
Fig. 1. Inhibitory effect of compound 1c on COX-2 protein expression. Raw 264.7 cells
(5 ꢂ 105) were incubated in 12 well culture plate for 24 h, and then treated with
Parameters
294
1.137
0.0696, 0.1968
0.1173, 0.2323
Goodness-of-fit on F2
compounds and LPS (1 mg/mL) for 18 h. After incubation, cells were lysed, and protein
R1, wR2 [I > 2
R1, wR2 [all data]
s
(I)]
was applied on SDSepolyacrylamide gel. The level of COX-2 protein expression was
examined by western blotting analysis.

226
R.-S. Yao et al. / European Journal of Medicinal Chemistry 62 (2013) 222e231
Table 4
5.2. Synthesis (Scheme 1)
ꢀ
ꢀ
Selected bond lengths (A) and angles ( ) for compound 3e.
5.2.1. Procedure for the preparation of (E)-3-(2,4-dimethoxy-6-((E)-
4-methoxystyryl)phenyl)acrylic acid (B)
Bond lengths
C16eC19
C20eC21
C10eC9
C8eN1
1.469(5)
1.464(5)
1.332(4)
1.344(5)
1.481(5)
1.363(4)
1.366(5)
1.411(5)
2.513(3)
C19eC20
C11eC10
C9eC8
C8eO1
C4eN1
C14eO3
C17eO2
H27eO1
H23/O2
1.326(5)
1.466(4)
1.477(4)
1.225(5)
1.470(5)
1.372(4)
1.426(5)
2.393(3)
2.625(3)
To a solution of (E)-2,4-dimethoxy-6-(4-methoxystyryl)benzal-
dehyde (A) (2.98 g, 10 mmol) and malonic acid (3.12 g, 30 mmol) in
100 mL of pyridine was added 2 mL of piperidine, and the reaction
mixture was stirred for 2 h at 95 ꢀC, followed by adjustment of
pH ¼ 2 with HCl solution. A yellow crude product was obtained and
purified by silica gel chromatography to afford the target product B.
Yellow solid; Yield: 92.1%; m.p. 150e152 ꢀC; 1H NMR (300 MHz,
C3eN1
C12eO2
C24eO4
C18eO3
H26/O1
CDCl3):
d
(ppm) 3.84 (s, 3H), 3.89 (s, 6H), 6.42 (d, 1H, J ¼ 2.4 Hz),
Bond angles
6.51 (d, 1H, J ¼ 15.9 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.89e6.94 (m, 3H),
7.27 (d, 1H, J ¼ 16.2 Hz), 7.47e7.48 (m, 2H), 8.13 (d, 1H, J ¼ 15.9 Hz).
C15eC16eC19
C16eC19eC20
C20eC21eC22
C16eC11eC10
C11eC10eC9
O1eC8eC9
118.8(3)
126.2(3)
122.6(3)
124.0(30)
128.4(3)
121.4(3)
157.0(2)
C11eC16eC19
C19eC20eC21
C20eC21eC26
C12eC11eC10
C10eC9eC8
121.2(3)
126.5(3)
120.3(3)
118.0(3)
120.4(3)
160.6(3)
158.8(3)
13C NMR (150 MHz, CDCl3):
d (ppm) 55.4, 97.5, 103.5, 114.2, 119.6,
124.8, 128.0, 132.1, 140.7, 160.7, 172.9. MS (ESI): 341.4 (C20H20O5,
[M þ H]þ). Anal. Calcd for C20H20O5: C, 70.57; H, 5.92%; Found: C,
75.42; H, 5.94%.
C27eH27BeO1
C23eH23/O2
C26eH26/O1
5.2.2. General procedure for the preparation of compounds 1aeq,
2aeh, 3ael
Compounds 1aeq, 2aeh, 3ael were synthesized by the known
procedure as previously reported [14].
The ESI-MS spectra were recorded on a Mariner System 5304 Mass
spectrometer. Carbon, hydrogen and nitrogen assays were carried
out with a CHNeO-Rapid instrument and were within ꢁ0.4% of the
theoretical values. TLC was run on the silica gel coated aluminum
sheets (Silica Gel 60 GF254, E. Merk, Germany) and visualized in
UV light (254 nm). Compound A was prepared as previously
reported.
5.2.2.1. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N-
ethylacrylamide (1a). White solid. Yield: 78.3%. m.p. 178e180 ꢀC 1H
NMR (CDCl3, 300 MHz):
d
1.18 (t, 3H, J ¼ 5.4 Hz), 3.40 (q, 2H,
Fig. 2. Crystal structure of compound 3e.

R.-S. Yao et al. / European Journal of Medicinal Chemistry 62 (2013) 222e231
227
Fig. 3. (a) Binding mode of compound 1c with COX-2; (b) Binding mode of compound
3e with COX-2. The hydrogen bonds were displayed as green dotted lines. Hydrophobic
interactions were shown as red balls (For interpretation of the references to color in
this figure legend, the reader is referred to the web version of this article.).
Fig. 4. (a) 3D mode of compound 1c binding with COX-2; (b) 3D mode of compound 3e
binding with COX-2. The protein was represented by surface and the compounds were
depicted by sticks and balls.
C24H29NO4: C, 72.89; H, 7.39; N, 3.54%; Found: C, 72.73; H, 7.41;
N, 3.55%.
J ¼ 5.4 Hz), 3.83 (s, 3H), 3.85 (s, 3H), 3.88 (s, 3H), 5.50 (bras,1H), 6.31
(d, 1H, J ¼ 11.7 Hz), 6.41 (d, 1H, J ¼ 1.8 Hz), 6.70 (d, 1H, J ¼ 1.8 Hz),
6.89e6.93 (m, 3H), 7.27 (d, 1H, J ¼ 12.0 Hz), 7.44e7.46 (m, 2H), 7.93
5.2.2.4. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N-
(d, 1H, J ¼ 11.7 Hz). 13C NMR (150 MHz, CDCl3):
d
(ppm) 14.9, 34.5,
hexylacrylamide (1d). White solid. Yield: 84.6%. m.p. 129e131 ꢀC 1H
97.6, 103.0, 114.1, 125.5, 127.9, 130.9, 134.6, 159.5, 166.6. MS (ESI):
368.4 (C22H25NO4, [M þ H]þ). Anal. Calcd for C22H25NO4: C, 71.91;
H, 6.86; N, 3.81%; Found: C, 71.73; H, 6.88; N, 3.82%.
NMR (CDCl3, 300 MHz):
d
0.88 (t, 3H, J ¼ 6.6 Hz), 1.26e1.36 (m, 6H),
1.49e1.57 (m, 2H), 3.33e3.39 (m, 2H), 3.83 (s, 3H), 3.85 (s, 3H), 3.88
(s, 3H), 5.53 (bras, 1H), 6.32 (d, 1H, J ¼ 15.6 Hz), 6.41 (d, 1H,
J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.88e6.93 (m, 3H), 7.28 (d, 1H,
J ¼ 16.2 Hz), 7.43e7.46 (m, 2H), 7.94 (d, 1H, J ¼ 15.6 Hz). 13C NMR
5.2.2.2. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N-
propylacrylamide (1b). White solid. Yield: 83.1%. m.p. 142e145 ꢀC
(150 MHz, CDCl3): d (ppm) 14.0, 22.5, 26.7, 29.6, 31.5, 39.7, 55.4,
1H NMR (CDCl3, 300 MHz):
d
0.95 (t, 3H, J ¼ 7.5 Hz), 1.54e1.63
97.6, 114.1, 128.0, 130.9, 134.6, 140.4, 160.1, 166.7. MS (ESI): 424.5
(C26H33NO4, [M þ H]þ). Anal. Calcd for C26H33NO4: C, 73.73; H, 7.85;
N, 3.31%; Found: C, 73.55; H, 7.88; N, 3.32%.
(m, 2H), 3.33 (q, 2H, J ¼ 6.6 Hz), 3.83 (s, 3H), 3.85 (s, 3H), 3.87 (s,
3H), 5.55 (bras, 1H), 6.33 (d, 1H, J ¼ 15.6 Hz), 6.40 (d, 1H, J ¼ 2.1 Hz),
6.70 (d, 1H, J ¼ 2.1 Hz), 6.88e6.93 (m, 3H), 7.28 (d, 1H, J ¼ 15.9 Hz),
7.44e7.46 (m, 2H), 7.94 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz,
5.2.2.5. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N-
octylacrylamide (1e). White solid. Yield: 78.4%. m.p. 142e134 ꢀC
CDCl3):
d
(ppm) 11.4, 22.9, 41.4, 55.3, 97.6, 103.0, 114.1, 124.2, 125.5,
127.9, 130.9, 134.6, 159.9, 166.7. MS (ESI): 382.5 (C23H27NO4,
[M þ H]þ). Anal. Calcd for C23H27NO4: C, 72.42; H, 7.13; N, 3.67%;
Found: C, 72.30; H, 7.15; N, 3.68%.
1H NMR (CDCl3, 300 MHz):
d
0.87 (t, 3H, J ¼ 6.6 Hz), 1.27e1.30
(m, 10H), 1.49e1.56 (m, 2H), 3.32e3.39 (m, 2H), 3.83 (s, 3H),
3.85 (s, 3H), 3.87 (s, 3H), 5.52 (bras, 1H), 6.32 (d, 1H, J ¼ 15.6 Hz),
6.41 (d, 1H, J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.87e6.93 (m, 3H),
7.28 (d, 1H, J ¼ 16.2 Hz), 7.43e7.46 (m, 2H), 7.93 (d, 1H,
5.2.2.3. (E)-N-butyl-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)
phenyl)acrylamide (1c). White solid. Yield: 87.2%. m.p. 153e155 ꢀC
J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 14.1, 22.6, 27.0,
1H NMR (CDCl3, 300 MHz):
d
0.93 (t, 3H, J ¼ 7.2 Hz), 1.34e1.41 (m,
29.2, 29.7, 31.8, 39.7, 55.6, 97.6, 114.1, 127.9, 130.9, 134.6, 140.4,
160.1, 166.7. MS (ESI): 452.6 (C28H37NO4, [M þ H]þ). Anal. Calcd
for C28H37NO4: C, 74.47; H, 8.26; N, 3.10%; Found: C, 74.32; H,
8.28; N, 3.11%.
2H), 1.49e1.56 (m, 2H), 3.34e3.40 (m, 2H), 3.83 (s, 3H), 3.85 (s,
3H), 3.88 (s, 3H), 5.49 (bras, 1H), 6.32 (d, 1H, J ¼ 15.6 Hz), 6.41 (d,
1H, J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.88e6.93 (m, 3H), 7.28
(d, 1H, J ¼ 16.2 Hz), 7.43e7.46 (m, 2H), 7.93 (d, 1H, J ¼ 15.6 Hz).
13C NMR (150 MHz, CDCl3):
d
(ppm) 13.7, 20.1, 31.7, 39.4, 55.4,
5.2.2.6. (E)-N-(2-chloroethyl)-3-(2,4-dimethoxy-6-((E)-4-methox-
97.6, 103.0, 114.1, 125.5, 127.9, 130.9, 134.7, 140.4, 159.4, 160.7,
ystyryl)phenyl)acrylamide (1f). Yellow solid. Yield: 81.9%. m.p. 146e
166.7. MS (ESI): 396.5 (C24H29NO4, [M þ H]þ). Anal. Calcd for
148 ꢀC 1H NMR (CDCl3, 300 MHz):
d 3.68e3.73 (m, 4H), 3.83

228
R.-S. Yao et al. / European Journal of Medicinal Chemistry 62 (2013) 222e231
(s, 3H), 3.86 (s, 3H), 3.88 (s, 3H), 5.93 (bras, 1H), 6.39 (d, 1H,
J ¼ 11.7 Hz), 6.42 (d, 1H, J ¼ 1.5 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.88e
6.93 (m, 3H), 7.27 (d, 1H, J ¼ 15.9 Hz), 7.44e7.47 (m, 2H), 7.97 (d, 1H,
134.7, 140.4, 159.9, 166.8. MS (ESI): 396.2 (C24H29NO4, [M þ H]þ).
Anal. Calcd for C24H29NO4: C, 72.89; H, 7.39; N, 3.54%; Found: C,
72.77; H, 7.37; N, 3.55%.
J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 29.7, 41.4, 44.2,
55.3, 97.6, 114.2, 123.2, 125.3, 127.9, 131.3, 135.6, 140.7, 160.1, 161.0,
166.9. MS (ESI): 402.1 (C22H24ClNO4, [M þ H]þ). Anal. Calcd for
C22H24ClNO4: C, 65.75; H, 6.02; N, 3.49%; Found: C, 65.91; H, 6.03;
N, 3.48%.
5.2.2.12. (E)-N-(sec-butyl)-3-(2,4-dimethoxy-6-((E)-4-methoxy-
styryl)phenyl)acrylamide (1l). White solid. Yield: 69.3%. m.p. 158e
160 ꢀC. 1H NMR (CDCl3, 300 MHz):
d
0.93 (t, 3H, J ¼ 7.2 Hz), 1.17
(d, 3H, J ¼ 6.6 Hz), 1.49e1.56 (m, 2H), 3.83 (s, 3H), 3.86 (s, 3H),
3.88 (s, 3H), 4.01e4.11 (m, 1H), 5.30 (bras, 1H), 6.33 (d, 1H,
J ¼ 15.6 Hz), 6.41 (d, 1H, J ¼ 2.1 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.88e
6.93 (m, 3H), 7.30 (d, 1H, J ¼ 15.9 Hz), 7.44e7.47 (m, 2H), 7.94 (d, 1H,
5.2.2.7. (E)-N-(3-chloropropyl)-3-(2,4-dimethoxy-6-((E)-4-
methoxystyryl)phenyl)acrylamide (1g). White solid. Yield: 65.3%.
m.p. 147e149 ꢀC 1H NMR (CDCl3, 300 MHz):
d
2.03e2.10 (m, 2H),
J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 10.4, 20.5, 29.7,
3.53 (t, 2H, J ¼ 6.3 Hz), 3.61 (t, 2H, J ¼ 6.3 Hz), 3.83 (s, 3H), 3.85 (s,
3H), 3.87 (s, 3H), 5.74 (bras, 1H), 6.36 (d, 1H, J ¼ 15.6 Hz), 6.41 (d,
1H, J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.87e6.92 (m, 3H), 7.27 (d,
1H, J ¼ 16.2 Hz), 7.43e7.46 (m, 2H), 7.95 (d, 1H, J ¼ 15.6 Hz). 13C
46.7, 55.3, 97.6, 103.0, 114.1, 127.9, 130.9, 134.6, 140.5, 160.7,
166.1. MS (ESI): 396.2 (C24H29NO4, [M þ H]þ). Anal. Calcd for
C24H29NO4: C, 72.89; H, 7.39; N, 3.54%; Found: C, 73.09; H, 7.38; N,
3.55%.
NMR (150 MHz, CDCl3):
d (ppm) 14.1, 22.7, 29.7, 32.2, 37.1, 42.6,
55.3, 97.5, 103.0, 114.1, 127.9, 131.0, 134.9, 140.6, 160.8, 167.2. MS
5.2.2.13. (E)-N-cyclopropyl-3-(2,4-dimethoxy-6-((E)-4-methox-
(ESI): 416.9 (C23H26ClNO4, [M
C23H26ClNO4: C, 66.42; H, 6.30; N, 3.37%; Found: C, 66.62; H, 6.32;
N, 3.36%.
þ
H]þ). Anal. Calcd for
ystyryl)phenyl)acrylamide (1m). White solid. Yield: 72.7%. m.p.
204e206 ꢀC. 1H NMR (CDCl3, 300 MHz):
d 0.52e0.58 (m, 2H), 0.78e
0.84 (m, 2H), 2.81e2.87 (m, 1H), 3.84 (s, 3H), 3.85 (s, 3H), 3.88 (s,
3H), 5.64 (bras, 1H), 6.28 (d, 1H, J ¼ 15.6 Hz), 6.40 (d, 1H, J ¼ 2.1 Hz),
6.70 (d, 1H, J ¼ 2.4 Hz), 6.87e6.93 (m, 3H), 7.27 (d, 1H, J ¼ 15.9 Hz),
5.2.2.8. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N-
(prop-2-yn-1-yl)acrylamide (1h). White solid. Yield: 67.8%. m.p.
7.43e7.47 (m, 2H), 7.96 (d, 1H,
J
¼
15.6 Hz). 13C NMR
172e174 ꢀC 1H NMR (CDCl3, 300 MHz):
d
2.22e2.24 (t, 1H, J ¼ 2.4),
(150 MHz, CDCl3):
d
(ppm) 45.9, 55.6, 97.6, 103.8, 136.6, 140.2, 160.7,
3.83 (s, 3H), 3.85 (s, 3H), 3.88 (s, 3H), 4.16e4.19 (m, 2H), 5.69 (bras,
1H), 6.37 (d, 1H, J ¼ 15.6 Hz), 6.41 (d, 1H, J ¼ 2.4 Hz), 6.70 (d, 1H,
J ¼ 2.4 Hz), 6.87e6.93 (m, 3H), 7.26 (d, 1H, J ¼ 15.9 Hz), 7.43e7.46
(m, 2H), 7.97 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
166.2. MS (ESI): 380.4 (C23H25NO4, [M
for C23H25NO4: C, 72.80; H, 6.64; N, 3.69%; Found: C, 72.72; H, 6.63;
N, 3.70%.
þ
H]þ). Anal. Calcd
d
(ppm) 29.3, 55.4, 97.6, 103.2, 114.2, 128.0, 131.3, 135.7, 140.7, 160.9,
5.2.2.14. (E)-N-cyclohexyl-3-(2,4-dimethoxy-6-((E)-4-methox-
166.5. MS (ESI): 378.4 (C23H23NO4, [M þ H]þ). Anal. Calcd for
C23H23NO4: C, 73.19; H, 6.14; N, 3.71%; Found: C, 73.38; H, 6.13; N,
3.70%.
ystyryl)phenyl)acrylamide (1n). White solid. Yield: 73.5%. m.p.187e
189 ꢀC. 1H NMR (CDCl3, 300 MHz):
d 1.04e1.25 (m, 4H), 1.33e
1.45 (m, 2H), 1.59e1.71 (m, 2H), 1.95e2.00 (m, 2H), 3.83 (s, 3H),
3.85 (s, 3H), 3.87 (s, 3H), 3.92e3.96 (m, 1H), 5.43 (bras, 1H), 6.32
(d, 1H, J ¼ 15.6 Hz), 6.40 (d, 1H, J ¼ 2.1 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz),
6.87e6.93 (m, 3H), 7.29 (d, 1H, J ¼ 16.50 Hz), 7.44e7.47 (m, 2H),
5.2.2.9. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N-
isopropylacrylamide (1i). White solid. Yield: 66.3%. m.p. 174e176 ꢀC
1H NMR (CDCl3, 300 MHz):
d
1.20 (d, 6H, J ¼ 3.6 Hz), 3.84 (s, 3H),
7.93 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 25.6,
3.86 (s, 3H), 3.88 (s, 3H), 4.17e4.28 (m,1H), 5.36 (bras, 1H), 6.31
(d, 1H, J ¼ 15.6 Hz), 6.41 (d, 1H, J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.1 Hz),
6.88e6.93 (m, 3H), 7.28 (d, 1H, J ¼ 15.6 Hz), 7.44e7.47 (m, 2H), 7.94
33.2, 48.3, 55.3, 97.6, 103.0, 125.4, 127.9, 130.9, 134.5, 140.4, 159.9,
165.7. MS (ESI): 422.5 (C26H31NO4, [M þ H]þ). Anal. Calcd for
C26H31NO4: C, 74.08; H, 7.41; N, 3.32%; Found: C, 74.26; H, 7.36;
N, 3.31%.
(d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 22.9, 41.4,
55.3, 97.6, 114.1, 128.0, 131.0, 134.6, 160.7, 165.9. MS (ESI): 382.5
(C23H27NO4, [M þ H]þ). Anal. Calcd for C23H27NO4: C, 72.42; H, 7.13;
N, 3.67%; Found: C, 72.26; H, 7.16; N, 3.68%.
5.2.2.15. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N-
(furan-2-ylmethyl)acrylamide (1o). White solid. Yield: 76.5%. m.p.
164e166 ꢀC. 1H NMR (CDCl3, 300 MHz):
d 3.77e3.94 (m, 9H), 4.55e
5.2.2.10. (E)-N-(tert-butyl)-3-(2,4-dimethoxy-6-((E)-4-methox-
4.59 (m, 2H), 5.85 (bras, 1H), 6.26e6.43 (m, 3H), 6.70e6.72 (m, 1H),
ystyryl)phenyl)acrylamide (1j). White solid. Yield: 65.9%. m.p.
6.88e6.93 (m, 3H), 7.25e7.48 (m, 5H), 7.96e8.04 (m, 1H). 13C NMR
145e147 ꢀC. 1H NMR (CDCl3, 300 MHz):
d
1.42 (s, 9H), 3.84 (s, 3H),
(150 MHz, CDCl3): d (ppm) 29.6, 36.6, 55.3, 97.6, 103.1, 107.4, 110.4,
3.86 (s, 3H), 3.87 (s, 3H), 5.36 (bras, 1H), 6.32 (d, 1H, J ¼ 15.6 Hz),
6.41 (d, 1H, J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.88e6.93 (m, 3H),
7.31 (d, 1H, J ¼ 15.9 Hz), 7.44e7.47 (m, 2H), 7.90 (d, 1H,
123.3, 125.3, 128.2, 128.0, 131.2, 135.4, 142.1, 151.4, 160.1, 160.8,
166.6. MS (ESI): 420.5 (C25H25NO5, [M þ H]þ). Anal. Calcd for
C25H25NO5: C, 71.58; H, 6.01; N, 3.34%; Found: C, 71.79; H, 5.99; N,
3.35%.
J ¼ 15.3 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 28.9, 51.3,
55.3, 97.6, 114.1, 125.4, 128.0, 131.0, 133.9, 140.5, 160.0, 166.2.
MS (ESI): 396.5 (C24H29NO4, [M
C24H29NO4: C, 72.89; H, 7.39; N, 3.54%; Found: C, 72.79; H, 7.41;
N, 3.55%.
þ
H]þ). Anal. Calcd for
5.2.2.16. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N-
(3-morpholinopropyl)acrylamide (1p). White solid. Yield: 74.9%.
m.p. 145e147 ꢀC 1H NMR (CDCl3, 300 MHz):
d 1.70e1.74 (m, 2H),
2.40e2.43 (m, 4H), 2.48 (t, 2H, J ¼ 6.0 Hz), 3.45e3.51 (m, 2H), 3.56e
3.59 (m, 4H), 3.83 (s, 3H), 3.86 (s, 3H), 3.88 (s, 3H), 6.30 (d, 1H,
J ¼ 15.6 Hz), 6.42 (d, 1H, J ¼ 2.1 Hz), 6.71(d, 1H, J ¼ 2.4 Hz), 6.88e
6.94 (m, 3H), 7.28 (d, 1H, J ¼ 15.9 Hz), 7.44e7.46 (m, 2H), 7.92 (d,
5.2.2.11. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N-
isobutylacrylamide (1k). White solid. Yield: 62.8%. m.p. 162e164 ꢀC.
1H NMR (CDCl3, 300 MHz):
d
0.94 (d, 6H, J ¼ 6.6 Hz), 1.79e1.88 (m,
1H), 3.20 (t, 2H, J ¼ 6.6 Hz), 3.83 (s, 3H), 3.86 (s, 3H), 3.88 (s, 3H),
5.57 (bras, 1H), 6.34 (d, 1H, J ¼ 15.6 Hz), 6.41 (d, 1H, J ¼ 2.4 Hz),
6.70 (d, 1H, J ¼ 2.4 Hz), 6.88e6.93 (m, 3H), 7.29 (d, 1H, J ¼ 16.2 Hz),
7.44e7.47 (m, 2H), 7.95 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz,
1H, J ¼ 15.9 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 24.5, 39.5, 53.4,
55.4, 57.8, 66.7, 97.6, 103.1, 114.1, 125.5, 127.9, 130.9, 134.3, 140.3,
159.9, 166.6. MS (ESI): 467.6 (C27H34N2O5, [M þ H]þ). Anal. Calcd for
C27H34N2O5: C, 69.51; H, 7.35; N, 6.00%; Found: C, 69.70; H, 7.36;
N, 5.98%.
CDCl3):
d (ppm) 20.1, 28.6, 47.0, 55.3, 97.6, 103.0, 114.1, 127.9, 130.8,

R.-S. Yao et al. / European Journal of Medicinal Chemistry 62 (2013) 222e231
229
5.2.2.17. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N-
6.86e6.93 (m, 3H), 7.28 (d, 1H, J ¼ 12.0 Hz), 7.42e7.45 (m, 2H), 8.00
(3,4,5-trimethoxyphenyl)acrylamide (1q). White solid. Yield: 65.3%.
(d, 1H, J ¼ 11.4 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 20.3, 27.0,
m.p. 155e157 ꢀC. 1H NMR (CDCl3, 300 MHz):
d
3.80 (s, 3H), 3.81 (s,
29.2, 55.2, 97.5, 103.2, 114.0, 116.6, 121.8, 125.9, 127.9, 129.9, 130.7,
135.7, 140.2, 160.6, 167.2. MS (ESI): 452.6 (C28H37NO4, [M þ H]þ).
Anal. Calcd for C28H37NO4: C, 74.47; H, 8.26; N, 3.10%; Found: C,
74.68; H, 8.24; N, 3.11%.
3H), 3.82 (s, 6H), 3.85 (s, 3H), 3.88 (s, 3H), 6.41 (d, 1H, J ¼ 2.1 Hz),
6.53 (d, 1H, J ¼ 15.6 Hz), 6.69 (d, 1H, J ¼ 2.4 Hz), 6.87e6.95 (m,
4H), 7.28 (d, 1H, J ¼ 15.9 Hz), 7.42e7.46 (m, 3H), 8.08 (d, 1H,
J ¼ 15.3 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 55.3, 56.5, 97.7,
103.2, 114.1, 125.3, 128.0, 129.9, 131.3, 133.5, 140.8, 159.5, 164.5. MS
(ESI): 506.6 (C29H31NO7, [M þ H]þ). Anal. Calcd for C29H31NO7: C,
68.90; H, 6.18; N, 2.77%; Found: C, 69.06; H, 6.17; N, 2.78%.
5.2.2.23. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N,N-
dipentylacrylamide (2f). Light white oil. Yield: 74.7%. 1H NMR
(CDCl3, 300 MHz):
d 0.71e0.92 (m, 12H), 1.05e1.61 (m, 6H), 2.99e
3.41 (m, 4H), 3.83 (s, 3H), 3.85 (s, 3H), 3.88 (s, 3H), 6.42 (d, 1H,
J ¼ 2.4 Hz), 6.70 (d,1H, J ¼ 1.5 Hz), 6.73e6.80 (m, 1H), 6.86e6.94 (m,
3H), 7.25e7.34 (m, 1H), 7.42e7.46 (m, 2H), 8.00 (d, 1H, J ¼ 15.6 Hz).
5.2.2.18. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N,N-
dimethylacrylamide (2a). Light white oil. Yield: 83.2%. 1H NMR
(CDCl3, 300 MHz):
d
3.04 (s, 6H), 3.83 (s, 3H), 3.85 (s, 3H), 3.88 (s,
13C NMR (150 MHz, CDCl3):
d (ppm) 11.4, 13.9, 16.7, 22.6, 27.1, 29.6,
3H), 6.42 (d, 1H, J ¼ 2.4 Hz), 6.71 (d, 1H, J ¼ 2.4 Hz), 6.81 (d, 1H,
J ¼ 15.6 Hz), 6.87e6.93 (m, 3H), 7.30 (d, 1H, J ¼ 16.2 Hz), 7.44e7.47
(m, 2H), 7.97 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
55.3, 97.6, 103.2, 114.0, 125.9, 127.9, 130.8, 159.4, 160.6. MS (ESI):
480.7 (C30H41NO4, [M þ H]þ). Anal. Calcd for C30H41NO4: C, 75.12;
H, 8.62; N, 2.92%; Found: C, 74.93; H, 8.61; N, 2.93%.
d
(ppm) 55.3, 55.6, 97.6, 103.1, 114.1, 116.1, 121.4, 125.8, 127.8, 130.7,
136.0, 140.2, 159.4, 160.6. MS (ESI): 368.4 (C22H25NO4, [M þ H]þ).
Anal. Calcd for C22H25NO4: C, 71.91; H, 6.86; N, 3.81%; Found: C,
72.07; H, 6.87; N, 3.80%.
5.2.2.24. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N,N-
dihexylacrylamide (2g). Light white oil. Yield: 79.1%. 1H NMR
(CDCl3, 300 MHz):
d
0.82 (t, 3H, J ¼ 7.2 Hz), 0.89 (t, 3H, J ¼ 6.9 Hz),
1.08e1.31 (m, 12H), 1.49e1.58 (m, 4H), 3.23 (t, 2H, J ¼ 7.50 Hz), 3.39
(t, 2H, J ¼ 7.5 Hz), 3.82 (s, 3H), 3.84 (s, 3H), 3.87 (s, 3H), 6.42 (d, 1H,
J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.76 (d, 1H, J ¼ 15.3 Hz), 6.86e
6.93 (m, 3H), 7.30 (d, 1H, J ¼ 16.2 Hz), 7.43e7.46 (m, 2H), 8.01 (d, 1H,
5.2.2.19. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N,N-
diethylacrylamide (2b). Light white oil. Yield: 81.3%. 1H NMR (CDCl3,
300 MHz):
d
1.07 (t, 3H, J ¼ 6.9 Hz), 1.17 (t, 3H, J ¼ 7.2 Hz), 3.34 (q,
2H, J ¼ 7.2 Hz), 3.47 (q, 2H, J ¼ 7.2 Hz), 3.81 (s, 3H), 3.84 (s, 3H), 3.87
(s, 3H), 6.41 (d, 1H, J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.76 (d, 1H,
J ¼ 15.6 Hz), 6.86e6.93 (m, 3H), 7.29 (d, 1H, J ¼ 15.9 Hz), 7.43e7.46
(m, 2H), 8.00 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 13.8, 22.5, 26.4,
26.7, 27.9, 29.7, 31.2, 31.6, 47.0, 48.2, 55.2, 97.5, 103.2, 114.0, 116.6,
121.7, 125.8, 127.8, 129.9, 130.7, 135.8, 140.2, 159.4, 160.6, 166.7. MS
(ESI): 508.7 (C32H45NO4, [M þ H]þ). Anal. Calcd for C32H45NO4: C,
75.70; H, 8.93; N, 2.76%; Found: C, 75.97; H, 8.91; N, 2.77%.
d
(ppm) 13.2, 40.9, 55.2, 97.6, 103.2, 114.0, 116.7, 121.8, 125.9, 127.8,
130.8, 135.8, 140.2, 159.8, 160.3, 166.4. MS (ESI): 396.5 (C24H29NO4,
[M þ H]þ). Anal. Calcd for C24H29NO4: C, 72.89; H, 7.39; N, 3.54%;
Found: C, 73.06; H, 7.41; N, 3.53%.
5.2.2.25. (E)-N,N-diallyl-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)
phenyl)acrylamide (2h). Light white oil. Yield: 75.6%. 1H NMR
(CDCl3, 300 MHz):
d 3.83 (s, 3H), 3.83 (s, 3H), 3.83e3.87 (m, 5H),
5.2.2.20. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N,N-
4.07e4.09 (m, 2H), 4.97e5.18 (m, 4H), 5.77e5.90 (m,1H), 5.59e5.71
(m, 1H), 6.41 (d, 1H, J ¼ 2.4 Hz), 6.69 (d, 1H, J ¼ 2.4 Hz), 6.73 (d, 1H,
J ¼ 15.3 Hz), 6.87e6.92 (m, 3H), 7.25 (d, 1H, J ¼ 15.9 Hz), 7.42e7.44
(m, 2H), 8.03 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
dipropylacrylamide (2c). Light white oil. Yield: 87.6%. 1H NMR
(CDCl3, 300 MHz):
d 0.68e0.93 (m, 6H), 1.55e1.64 (m, 4H), 3.24e
3.37 (m, 4H), 3.83 (s, 3H), 3.85 (s, 3H), 3.88 (s, 3H), 6.42 (d, 1H,
J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.77 (d, 1H, J ¼ 15.3 Hz), 6.86e
6.93 (m, 3H), 7.31 (d, 1H, J ¼ 15.9 Hz), 7.43e7.48 (m, 2H), 8.03 (d, 1H,
d
(ppm) 48.7, 55.6, 97.6, 103.2, 114.0, 116.5, 117.1, 121.3, 125.8, 127.9,
130.9, 133.5, 136.7, 140.3, 159.4, 160.7, 167.4. MS (ESI): 420.5
(C26H29NO4, [M þ H]þ). Anal. Calcd for C26H29NO4: C, 74.44; H, 6.97;
N, 3.34%; Found: C, 74.22; H, 6.99; N, 3.35%.
J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 10.9, 22.1, 22.9,
48.5, 49.8, 55.2, 97.4, 103.2, 109.3, 113.9, 116.5, 121.6, 125.8, 127.8,
130.7, 135.7, 140.2, 160.6, 166.7. MS (ESI): 424.5 (C26H33NO4,
[M þ H]þ). Anal. Calcd for C26H33NO4: C, 73.73; H, 7.85; N, 3.31%;
Found: C, 73.89; H, 7.88; N, 3.30%.
5.2.2.26. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-1-
(piperidin-1-yl)prop-2-en-1-one (3a). White solid. Yield: 83.7%.
m.p. 92e93 ꢀC. 1H NMR (CDCl3, 300 MHz):
d 1.50e1.66 (m, 6H),
5.2.2.21. (E)-N,N-dibutyl-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)
3.45 (bras, 2H), 3.65 (bras, 2H), 3.82 (s, 3H), 3.84 (s, 3H), 3.87 (s,
3H), 6.41 (d, 1H, J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.80 (d, 1H,
J ¼ 15.6 Hz), 6.87e6.93 (m, 3H), 7.28 (d, 1H, J ¼ 16.2 Hz), 7.43e7.46
(m, 2H), 7.94 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
phenyl)acrylamide (2d). Light white oil. Yield: 85.4%. 1H NMR
(CDCl3, 300 MHz):
d
0.75 (t, 3H, J ¼ 5.4 Hz), 0.94 (t, 3H, J ¼ 5.4 Hz),
1.09e1.15 (m, 2H), 1.32e1.37 (m, 2H), 1.46e1.59 (m, 4H), 3.24 (t, 2H,
J ¼ 5.7 Hz), 3.40 (t, 2H, J ¼ 5.7 Hz), 3.81 (s, 3H), 3.84 (s, 3H), 3.86 (s,
3H), 6.41 (d, 1H, J ¼ 1.8 Hz), 6.70 (d, 1H, J ¼ 1.5 Hz), 6.78 (d, 1H,
J ¼ 11.4 Hz), 6.86e6.93 (m, 3H), 7.30 (d, 1H, J ¼ 12.0 Hz), 7.43e7.45
d
(ppm) 24.6, 55.3, 97.6, 103.0, 114.1, 121.5, 125.9, 127.8, 130.6, 136.0,
140.1, 160.6, 166.1. MS (ESI): 408.5 (C25H29NO4, [M þ H]þ). Anal.
Calcd for C25H29NO4: C, 73.68; H, 7.17; N, 3.44%; Found: C, 73.49; H,
7.20; N, 3.43%.
(m, 2H), 8.01 (d,1H, J ¼ 11.7 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm)
13.5, 19.9, 30.1, 46.7, 55.2, 97.6, 103.2, 114.0, 116.7, 121.7, 125.9, 127.9,
135.8, 140.2, 160.6, 166.7. MS (ESI): 452.3 (C28H37NO4, [M þ H]þ).
Anal. Calcd for C28H37NO4: C, 74.47; H, 8.26; N, 3.10%; Found: C,
74.62; H, 8.28; N, 3.09%.
5.2.2.27. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-1-
(2-methylpiperidin-1-yl)prop-2-en-1-one (3b). Light white oil.
Yield: 65.2%. 1H NMR (CDCl3, 300 MHz):
d
1.16 (d, 3H, J ¼ 6.9 Hz),
1.39e1.64 (m, 7H), 3.83 (s, 3H), 3.85 (s, 3H), 3.88 (s, 3H), 6.41 (d, 1H,
J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.78 (d, 1H, J ¼ 15.6 Hz), 6.87e
6.93 (m, 3H), 7.28 (d, 1H, J ¼ 16.2 Hz), 7.43e7.46 (m, 2H), 7.94 (d, 1H,
5.2.2.22. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-N,N-
diisobutylacrylamide (2e). Light white oil. Yield: 72.2%. 1H NMR
(CDCl3, 300 MHz):
d
0.72 (d, 6H, J ¼ 5.1 Hz), 0.90 (d, 6H, J ¼ 5.1 Hz),
J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 18.8, 55.3, 97.5,
1.82e1.89 (m, 1H), 2.04e2.11 (m, 1H), 3.09 (d, 2H, J ¼ 5.7 Hz),
3.28 (d, 2H, J ¼ 5.7 Hz), 3.82 (s, 3H), 3.85 (s, 3H), 3.87 (s, 3H), 6.41
(d, 1H, J ¼ 1.8 Hz), 6.70 (d, 1H, J ¼ 1.8 Hz), 6.80 (d, 1H, J ¼ 11.7 Hz),
103.1, 114.0, 116.7, 122.0, 127.8, 130.5, 135.7, 139.9, 159.7, 160.5, 166.2.
MS (ESI): 422.5 (C26H31NO4, [M þ H]þ). Anal. Calcd for C26H31NO4:
C, 74.08; H, 7.41; N, 3.32%; Found: C, 74.35; H, 7.43; N, 3.31%.

230
R.-S. Yao et al. / European Journal of Medicinal Chemistry 62 (2013) 222e231
5.2.2.28. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-1-
2H), 1.69e1.80 (m, 2H), 2.50e2.96 (m, 4H), 3.82 (s, 3H), 3.85 (s, 3H),
3.87 (s, 3H), 4.49e4.74 (m, 1H), 6.41 (d, 1H, J ¼ 2.4 Hz), 6.70 (d, 1H,
J ¼ 2.1 Hz), 6.80 (d,1H, J ¼ 15.3 Hz), 6.86e6.93 (m, 3H), 7.11e7.14 (m,
2H), 7.21e7.25 (m, 1H), 7.25e7.31 (m, 3H), 7.42e7.45 (m, 2H), 7.94
(3-methylpiperidin-1-yl)prop-2-en-1-one (3c). White solid. Yield:
71.4%. m.p. 115e117 ꢀC. 1H NMR (CDCl3, 300 MHz):
d 0.71e0.93 (m,
3H), 1.10e1.82 (m, 6H), 2.28e2.97 (m, 2H), 3.83 (s, 3H), 3.85 (s, 3H),
3.88 (s, 3H), 4.46e4.57 (m, 1H), 6.41 (d, 1H, J ¼ 2.4 Hz), 6.71 (d, 1H,
J ¼ 2.4 Hz), 6.80 (d, 1H, J ¼ 15.3 Hz), 6.87e6.93 (m, 3H), 7.28 (d, 1H,
J ¼ 16.2 Hz), 7.43e7.46 (m, 2H), 7.94 (d, 1H, J ¼ 15.1 Hz). 13C NMR
(d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 38.3, 42.9,
55.3, 97.6, 103.1, 114.1, 125.1, 128.2, 136.1, 140.0, 160.6, 166.0. MS
(ESI): 484.6 (C31H33NO4, [M þ H]þ). Anal. Calcd for C31H33NO4: C,
76.99; H, 6.88; N, 2.90%; Found: C, 76.87; H, 6.90; N, 2.91%.
(150 MHz, CDCl3):
d (ppm) 33.1, 55.4, 97.6, 103.1, 114.1, 121.7, 127.8,
130.6, 135.9, 140.1, 160.6, 166.0. MS (ESI): 422.2 (C26H31NO4,
[M þ H]þ). Anal. Calcd for C26H31NO4: C, 74.08; H, 7.41; N, 3.32%;
Found: C, 73.97; H, 7.40; N, 3.34%.
5.2.2.34. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-1-
(4-methylpiperazin-1-yl)prop-2-en-1-one (3i). White solid. Yield:
84.7%. m.p. 73e74 ꢀC. 1H NMR (CDCl3, 300 MHz):
d 2.29 (s, 3H),
5.2.2.29. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-1-
(4-methylpiperidin-1-yl)prop-2-en-1-one (3d). Light white oil.
2.36e2.41 (m, 4H), 3.54e3.76 (m, 4H), 3.83 (s, 3H), 3.85 (s, 3H),
3.88 (s, 3H), 6.42 (d, 1H, J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.1 Hz), 6.79 (d,
1H, J ¼ 15.6 Hz), 6.87e6.93 (m, 3H), 7.26 (d, 1H, J ¼ 16.2 Hz), 7.43e
7.46 (m, 2H), 7.96 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
Yield: 74.5%. 1H NMR (CDCl3, 300 MHz):
d
0.93 (d, 3H, J ¼ 6.3 Hz),
1.07e1.17 (m, 2H), 1.56e1.76 (m, 4H), 2.60e2.68 (m, 1H), 2.93e3.01
(m, 1H), 3.83 (s, 3H), 3.85 (s, 3H), 3.88 (s, 3H), 4.66e4.71 (m, 1H),
6.41 (d, 1H, J ¼ 2.4 Hz), 6.71 (d, 1H, J ¼ 2.4 Hz), 6.80 (d, 1H,
J ¼ 15.3 Hz), 6.87e6.93 (m, 3H), 7.28 (d, 1H, J ¼ 16.2 Hz), 7.43e7.46
(m, 2H), 7.94 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
d
(ppm) 6.7, 22.8, 55.3, 97.6, 103.0, 114.1, 123.6, 125.4, 127.9, 131.0,
134.8, 140.5, 159.4, 168.1. MS (ESI): 423.5 (C25H30N2O4, [M þ H]þ).
Anal. Calcd for C25H30N2O4: C, 71.07; H, 7.16; N, 6.63%; Found: C,
70.91; H, 7.15; N, 6.65%.
d
(ppm) 21.7, 31.1, 55.3, 97.6, 103.1, 114.1, 116.8, 121.7, 125.9, 127.8,
130.6, 135.9, 140.0, 160.6, 166.0. MS (ESI): 422.3 (C26H31NO4,
[M þ H]þ). Anal. Calcd for C26H31NO4: C, 74.08; H, 7.41; N, 3.32%;
Found: C, 73.99; H, 7.39; N, 3.32%.
5.2.2.35. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-1-
morpholinoprop-2-en-1-one (3j). White solid. Yield: 88.2%. m.p.
103e105 ꢀC. 1H NMR (CDCl3, 300 MHz):
d 3.45e3.69 (m, 8H), 3.83
(s, 3H), 3.86 (s, 3H), 3.88 (s, 3H), 6.42 (d, 1H, J ¼ 2.4 Hz), 6.70 (d, 1H,
J ¼ 2.4 Hz), 6.77 (d, 1H, J ¼ 15.6 Hz), 6.88e6.93 (m, 3H), 7.26 (d, 1H,
J ¼ 15.9 Hz), 7.42e7.45 (m, 2H), 7.99 (d, 1H, J ¼ 15.6 Hz). 13C NMR
5.2.2.30. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-1-
(3,5-dimethylpiperidin-1-yl)prop-2-en-1-one (3e). White solid.
Yield: 76.1%. m.p. 118e120 ꢀC. 1H NMR (CDCl3, 300 MHz):
d
0.69e
(150 MHz, CDCl3): d (ppm) 29.6, 36.5, 39.5, 55.3, 66.8, 97.6, 103.2,
0.91 (m, 8H), 1.53e1.62 (m, 2H), 1.77e1.91 (m, 2H), 2.03e2.09 (m,
1H), 2.41e2.47 (m,1H), 3.82 (s, 3H), 3.85 (s, 3H), 3.87 (s, 3H), 6.41 (d,
1H, J ¼ 1.5 Hz), 6.70 (d,1H, J ¼ 1.8 Hz), 6.79 (d,1H, J ¼ 11.7 Hz), 6.87e
6.93 (m, 3H), 7.26 (d, 1H, J ¼ 12.0 Hz), 7.43e7.46 (m, 2H), 7.93 (d, 1H,
114.1, 120.2, 125.8, 127.8, 130.9, 136.9, 140.4, 160.8, 166.4. MS (ESI):
410.5 (C24H27NO5, [M þ H]þ). Anal. Calcd for C24H27NO5: C, 70.40;
H, 6.65; N, 3.42%; Found: C, 70.23; H, 6.66; N, 3.43%.
J ¼ 11.7 Hz). 13C NMR (150 MHz, CDCl3):
d
(ppm) 19.0, 42.5, 55.4,
5.2.2.36. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-1-
97.6, 103.1, 114.1, 121.6, 126.0, 127.8, 130.6, 136.0, 140.1, 159.7, 160.6,
165.8. MS (ESI): 436.6 (C27H33NO4, [M þ H]þ). Anal. Calcd for
C27H33NO4: C, 74.45; H, 7.64; N, 3.22%; Found: C, 74.33; H, 7.67;
N, 3.21%.
thiomorpholinoprop-2-en-1-one (3k). White solid. Yield: 83.6%.
m.p. 138e140 ꢀC. 1H NMR (CDCl3, 300 MHz):
d 2.48e2.69 (m, 4H),
3.77e3.82 (m, 4H), 3.83 (s, 3H), 3.86 (s, 3H), 3.88 (s, 3H), 6.42 (d,
1H, J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz), 6.75 (d, 1H, J ¼ 15.3 Hz),
6.88e6.93 (m, 3H), 7.25 (d, 1H, J ¼ 16.2 Hz), 7.43e7.46 (m, 2H),
5.2.2.31. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-1-
7.98 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz, CDCl3):
d (ppm) 55.3,
(2-ethylpiperidin-1-yl)prop-2-en-1-one (3f). White solid. Yield:
97.6, 103.2, 114.1, 120.7, 125.8, 127.8, 130.9, 136.9, 140.3, 160.8,
166.4. MS (ESI): 426.5 (C24H27NO4S, [M þ H]þ). Anal. Calcd for
C24H27NO4S: C, 67.74; H, 6.40; N, 3.29%; Found: C, 67.63; H, 6.41;
N, 3.30%.
77.4%. m.p. 91e92 ꢀC. 1H NMR (CDCl3, 300 MHz):
d 0.68e0.90 (m,
3H), 1.46e1.77 (m, 9H), 2.61e3.03 (m, 1H), 3.83 (s, 3H), 3.85 (s, 3H),
3.88 (s, 3H), 4.62e4.84 (m, 1H), 6.42 (d, 1H, J ¼ 2.1 Hz), 6.70 (d, 1H,
J ¼ 2.4 Hz), 6.79 (d, 1H, J ¼ 15.6 Hz), 6.86e6.93 (m, 3H), 7.28 (d, 1H,
J ¼ 15.9 Hz), 7.42e7.45 (m, 2H), 7.94 (d, 1H, J ¼ 15.6 Hz). 13C NMR
5.2.2.37. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-1-
(150 MHz, CDCl3):
d
(ppm) 10.7, 19.1, 55.3, 97.6, 103.0, 114.0, 127.8,
(pyrrolidin-1-yl)prop-2-en-1-one (3l). White solid. Yield: 76.7%.
130.5, 135.7, 160.5. MS (ESI): 436.4 (C27H33NO4, [M þ H]þ). Anal.
Calcd for C27H33NO4: C, 74.45; H, 7.64; N, 3.22%; Found: C, 74.71; H,
7.62; N, 3.23%.
m.p. 100e101 ꢀC. 1H NMR (CDCl3, 300 MHz):
d 1.84e1.93 (m, 4H),
3.45 (t, 2H, J ¼ 6.6 Hz), 3.59 (t, 2H, J ¼ 6.6 Hz), 3.83 (s, 3H), 3.85 (s,
3H), 3.88 (s, 3H), 6.42 (d, 1H, J ¼ 2.1 Hz), 6.64 (d, 1H, J ¼ 15.6 Hz),
6.71 (d, 1H, J ¼ 2.4 Hz), 6.88e6.94 (m, 3H), 7.29 (d, 1H, J ¼ 15.9 Hz),
7.44e7.46 (m, 2H), 7.99 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz,
5.2.2.32. (E)-1-(4-chloropiperidin-1-yl)-3-(2,4-dimethoxy-6-((E)-4-
methoxystyryl)phenyl)prop-2-en-1-one (3g). White solid. Yield:
CDCl3): d (ppm) 24.3, 26.1, 46.4, 97.6, 103.0, 114.1, 122.8, 125.8, 127.9,
72.8%. m.p. 61e62 ꢀC. 1H NMR (CDCl3, 300 MHz):
d
1.77e1.86 (m,
130.6, 135.3, 140.2, 160.6, 165.5. MS (ESI): 394.5 (C24H27NO4,
[M þ H]þ). Anal. Calcd for C24H27NO4: C, 73.26; H, 6.92; N, 3.56%;
Found: C, 73.45; H, 6.91; N, 3.57%.
2H), 2.01e2.07 (m, 2H), 3.52e3.59 (m, 2H), 3.79e3.88 (m, 11H),
4.24e4.29 (m, 1H), 6.42 (d, 1H, J ¼ 2.4 Hz), 6.70 (d, 1H, J ¼ 2.4 Hz),
6.80 (d, 1H, J ¼ 15.3 Hz), 6.88e6.93 (m, 3H), 7.27 (d, 1H, J ¼ 15.9 Hz),
7.43e7.46 (m, 2H), 7.97 (d, 1H, J ¼ 15.6 Hz). 13C NMR (150 MHz,
5.3. X-ray crystallography
CDCl3):
d (ppm) 55.4, 56.9, 97.6, 103.2, 114.1, 120.6, 125.8, 127.8,
130.9, 136.8, 140.3, 160.8, 166.2. MS (ESI): 442.9 (C25H28ClNO4,
[M þ H]þ). Anal. Calcd for C25H28ClNO4: C, 67.94; H, 6.39; N, 3.17%;
Found: C, 67.83; H, 6.37; N, 3.18%.
The crystallographic data for compound 3e were collected on
a Bruker Smart 1000 CCD area detector diffractometer. Equipped
ꢀ
with Mo K
a
(l
¼ 0.71073A) radiation using
u-scan mode. Empirical
absorption correction was applied to the data. The structures were
solved by direct methods and refined by full-matrix least-squares
methods on F2. All non-hydrogen atoms were located from the trial
structure and then refined anisotropically. All hydrogen atoms were
5.2.2.33. (E)-3-(2,4-dimethoxy-6-((E)-4-methoxystyryl)phenyl)-1-
(4-phenylpiperidin-1-yl)prop-2-en-1-one (3h). White solid. Yield:
67.4%. m.p. 127e129 ꢀC. 1H NMR (CDCl3, 300 MHz):
d 1.11e1.24 (m,

R.-S. Yao et al. / European Journal of Medicinal Chemistry 62 (2013) 222e231
231
generated in idealized positions and were assigned fixed isotropic
Appendix A. Supplementary material
thermal parameters at 1.2 times the equivalent isotropic U of the
atoms to which they are attached and allowed to ride on their
respective parent atoms. The contributions of these hydrogen
atoms were included in the structure-factors calculations.
Other relevant parameters of the crystal structure are listed in
CCDC 845819 contains the supplementary crystallographic data
for compound 3e. These data can be obtained free of charge from
The Cambridge Crystallographic Data Centre via www.ccdc.cam.ac.
uk/data_request/cif.
ꢀ
Table 3. Table 4 shows some selected bond lengths (A) and angles
for 3e.
References
5.4. Antiproliferation assay
[1] C.D. Funk, Prostaglandins and leukotrienes: advances in eicosanoid biology,
Science 294 (2001) 1871e1875.
[2] S. Kase, M. Osaki, S. Honjo, K. Hashimoto, H. Adachi, S. Tsujitani, H. Ito,
Expression of cyclooxygenase-1 and cyclooxygenase-2 in human esophageal
mucosa, dysplasia and carcinoma, Pathobiology 71 (2004) 84e92.
[3] O. Laneuville, D.K. Breuer, D.L. Dewitt, T. Hla, C.D. Funk, W.L. Smith, Differ-
ential inhibition of human prostaglandin endoperoxide H synthases-1 and -2
by nonsteroidal anti-inflammatory drugs, J. Pharmacol. Exp. Ther. 271 (1994)
927e934.
[4] H.X. Li, X.M. Chang, Z.J. Song, S.X. He, Correlation between expression of
cyclooxygenase-2 and angiogenesis in human gastric adenocarcinoma, World
J. Gastroenterol. 9 (2003) 674e677.
The antiproliferative activity of the prepared compounds
against B16-F10, MCF-7 and A549 cell lines was evaluated as
described elsewhere with some modifications [17]. Target tumor
cell lines were grown to log phase in RPMI 1640 medium supple-
mented with 10% fetal bovine serum. After diluting to
2 ꢂ 104 cells mLꢃ1 with the complete medium, 100
mL of the ob-
tained cell suspension was added to each well of 96-well culture
plates. The subsequent incubation was permitted at 37 ꢀC, 5% CO2
atmosphere for 24 h before the cytotoxicity assessments. Tested
samples at pre-set concentrations were added to 6 wells with
celecoxib coassayed as positive reference. After 48 h exposure
[5] W. Dempke, C. Rie, A. Grothey, H.J. Schmoll, Cyclooxygenase-2: a novel target
for cancer chemotherapy? J. Cancer Res. Clin. Oncol. 127 (2001) 411e417.
ꢁ
[6] G. Eren, S. Ünlü, M.T. Nuñz, L. Labeaga, F. Ledo, A. Entrena, E. Banoglu,
G. Costantino, M.F. S¸ ahin, Synthesis, biological evaluation, and docking studies
of novel heterocyclic diaryl compounds as selective COX-2 inhibitors, Bioorg.
Med. Chem. 18 (2010) 6367e6376.
period, 40
dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)) was
added to each well. 4 h later, 100 L extraction solution (10% SDSe
m
L of PBS containing 2.5 mg mLꢃ1 of MTT (3-(4,5-
[7] F. Julémont, X.D. Leval, C. Michaux, J.F. Renard, J.Y. Winum, J.L. Montero,
J. Damas, J.M. Dogné, B. Pirotte, Design, synthesis, and pharmacological eval-
uation of pyridinic analogues of nimesulide as cyclooxygenase-2 selective
inhibitors, J. Med. Chem. 47 (2004) 6749e6759.
[8] N. Pommery, T. Taverne, A. Telliez, L. Goossens, C. Charlier, J. Pommery,
J.F. Goossens, R. Houssin, F. Durant, J.P. Hénichart, New COX-2/5-LOX inhibi-
tors: apoptosis-inducing agents potentially useful in prostate cancer chemo-
therapy, J. Med. Chem. 47 (2004) 6195e6206.
[9] F.A. Attiga, P.M. Fernandez, A.T. Weeraratna, M.J. Manyak, S.R. Patierno,
Inhibitors of prostaglandin synthesis inhibit human prostate tumor cell
invasiveness and reduce the release of matrix metalloproteinases, Cancer Res.
60 (2000) 4629e4637.
m
5% isobutyl alcohole0.01 M HCl) was added. After an overnight
incubation at 37 ꢀC, the optical density was measured at a wave-
length of 570 nm on an ELISA microplate reader. In all experiments
three replicate wells were used for each drug concentration. Each
assay was carried out at least three times. The results were
summarized in Table 2.
5.5. In vitro cyclooxygenase (COX) inhibition assays
[10] M.V.R. Reddy, V.K. Billa, V.R. Pallela, M.R. Mallireddigari, R. Boominathan,
J.L. Gabriel, E.P. Reddy, Design, synthesis, and biological evaluation of 1-(4-
sulfamylphenyl)-3-trifluoromethyl-5-indolyl pyrazolines as cyclooxygenase-
2 (COX-2) and lipoxygenase (LOX) inhibitors, Bioorg. Med. Chem. 16 (2008)
3907e3916.
[11] N.M. Davies, F. Jamali, COX-2 selective inhibitors cardiac toxicity: getting to
the heart of the matter, J. Pharm. Pharm. Sci. 7 (2004) 332e336.
[12] K. Likhitwitayawuid, K. Sawasdee, K. Kirtikara, Flavonoids and stilbenoids
with COX-1 and COX-2 inhibitory activity from Dracaena loureiri, Planta Med.
68 (2002) 841e843.
[13] M. Murias, N. Handler, T. Erker, K. Pleban, G. Ecker, P. Saiko, T. Szekeres,
W. Jäger, Resveratrol analogues as selective cyclooxygenase-2 inhibitors:
synthesis and structureeactivity relationship, Bioorg. Med. Chem. 12 (2004)
5571e5578.
[14] B.F. Ruan, X. Lu, J.F. Tang, Y. Wei, X.L. Wang, Y.B. Zhang, L.S. Wang, H.L. Zhu,
Synthesis, biological evaluation, and molecular docking studies of resveratrol
derivatives possessing chalcone moiety as potential antitubulin agents, Bio-
org. Med. Chem. 19 (2011) 2688e2695.
The ability of the test compounds to inhibit COX-1 and COX-2
was determined using chemiluminescent enzyme assays kit
(Cayman Chemical, Ann Arbor, MI, USA) according to previously
reported method [18].
5.6. Anti-inflammatory assay
The inhibitory effects of samples on PGE2 expression were
evaluated in lipopolysaccharide (LPS)-activated murine macro-
phage RAW 264.7 cells, using a method modified from that previ-
ously reported [19].
[15] I.M. El-Deeb, S.H. Lee, Design and synthesis of new anticancer pyrimidines with
multiple-kinase inhibitory effect, Bioorg. Med. Chem. 18 (2010) 3860e3874.
[16] X.F. Huang, L. Shi, H.Q. Li, H.L. Zhu, Synthesis and crystal structure of 4,6-
dihydroxy-2-[2-(4-hydroxyphenyl)-vinyl]-benzene-1,3-dicarbaldehyde,
J. Chem. Crystallogr. 37 (2007) 739e742.
[17] A. Boumendjel, J. Boccard, P.A. Carrupt, E. Nicolle, M. Blanc, A. Geze,
L. Choisnard, D. Wouessidjewe, E.L. Matera, C. Dumontet, Antimitotic and
antiproliferative activities of chalcones: forward structureeactivity relation-
ship, J. Med. Chem. 51 (2008) 2307e2310.
5.7. Western immunoblot analysis
RAW 264.7 cells were pretreated with 1c at the concentrations
of 0.5, 1.0, 1.5 mM for 15 min before treatment with 1 m
g mLꢃ1 LPS
for 18 h and examining the expression of COX-2 protein. Cells were
lysed with lysis buffer. Western immunoblot analysis was per-
formed using a method described by Cheenpracha et al. [20].
[18] A.G.E. Amr, M.M. Abdulla, Anti-inflammatory profile of some synthesized
heterocyclic pyridone and pyridine derivatives fused with steroidal structure,
Bioorg. Med. Chem. 14 (2006) 4341e4352.
5.8. Molecular modeling (docking) studies
[19] C.K. Lii, H.W. Chen, W.T. Yun, K.L. Liu, Suppressive effects of wild bitter gourd
(Momordica charantia Linn. var. abbreviate ser.) fruit extracts on inflamma-
tory responses in RAW264.7 macrophages, J. Ethnopharmacol. 122 (2009)
227e233.
[20] S. Cheenpracha, E.J. Park, W.Y. Yoshida, C. Barit, M. Wall, J.M. Pezzuto,
L.C. Chang, Potential anti-inflammatory phenolic glycosides from the medic-
inal plant Moringa oleifera fruits, Bioorg. Med. Chem. 18 (2010) 6598e6602.
[21] R. Huey, G.M. Morris, A.J. Olson, D.S. Goodsell, A semiempirical free energy
force field with charge-based desolvation, J. Comput. Chem. 28 (2007) 1145e
1152.
Molecular docking of compounds into the three dimensional
COX-2 complex structure (PDB code: 1cx2) was carried out using
the Molsoft ICM-Pro software package (version 3.5-0a) [21,22].
Acknowledgments
This work was supported by the Fundamental Research Funds
for the Central Universities (2012HGZY0021, 2012HGCX0003 and
2012HGQC0034).
[22] F. Musiani, E. Arnofi, R. Casadio, S. Ciurli, Structure-based computational study
of the catalytic and inhibition mechanisms of urease, J. Biol. Inorg. Chem. 6
(2001) 300e314.