Synthesis and biological evaluation of novel hybrid chalcone derivatives as vasorelaxant agents

Article abstract of DOI:10.1016/j.ejmech.2010.05.054

With the aim to further improve the vasorelaxant activities of chalcones, nine hybrid chalcone derivatives conjugated with nitric oxide (NO) donor or 1,4-dihydropyridyl (1,4-DHP) moiety were designed and synthesized based on molecular hybridization strategy. Their vasorelaxant activities were evaluated in aortic rings with endothelium pre-contracted with phenylephrine (PE). All of these compounds showed preferable vasorelaxant activities which were more potent than their parent compounds. Compounds 8c and 15c, the most potent compounds, would be promising structural templates for the development of novel vasorelaxant agents. Nine hybrid chalcone derivatives conjugated with nitric oxide (NO) donors or 1,4-dihydropyridyl moiety were designed, synthesized and biological evaluated for their vasorelaxant activities.

Full text of DOI:10.1016/j.ejmech.2010.05.054

European Journal of Medicinal Chemistry 45 (2010) 3986e3992
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis and biological evaluation of novel hybrid chalcone derivatives as
vasorelaxant agents
a
a
a
a
b
a,
*
Xiaowu Dong , Lilin Du , Zhichao Pan , Tao Liu , Bo Yang , Yongzhou Hu
a
ZJU-ENS Joint Laboratory of Medicinal Chemistry, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China
Institute of Pharmacology & Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China
b
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 19 January 2010
Received in revised form
With the aim to further improve the vasorelaxant activities of chalcones, nine hybrid chalcone deriva-
tives conjugated with nitric oxide (NO) donor or 1,4-dihydropyridyl (1,4-DHP) moiety were designed and
synthesized based on molecular hybridization strategy. Their vasorelaxant activities were evaluated in
aortic rings with endothelium pre-contracted with phenylephrine (PE). All of these compounds showed
preferable vasorelaxant activities which were more potent than their parent compounds. Compounds 8c
and 15c, the most potent compounds, would be promising structural templates for the development of
novel vasorelaxant agents.
25 May 2010
Accepted 26 May 2010
Available online 2 June 2010
Keywords:
Hybrid chalcones
Nitric oxide donor
Ó 2010 Elsevier Masson SAS. All rights reserved.
1,4-Dihydropyridyl moiety
Vasorelaxant agent
1. Introduction
exhibited favorable vasorelaxant activities and the hydroxyl group
at the 2 position of chalcones was an interesting point for further
Flavonoids are a group of naturally occurring phenolic phyto-
chemicals with diverse pharmacological properties, such as anti-
bacterial, anti-viral, anti-allergic, anti-hepatotoxic, anti-ulcer action
and anti-cancer [1e3]. Recently, they have been recognized as
compounds with potent biological activities that may be active in
prevention of chronic diseases such as cardiovascular disease [4].
Flavonoids showed beneficial impact on parameters associated
with atherosclerosis, including lipoprotein oxidation, platelet
aggregation and vascular reactivity [5e7]. Also, the vasorelaxant
activities of flavonoids were extensively studied in isolated vascular
preparation and animal models [8,9], suggesting that their vaso-
relaxant mechanisms were involved with the modulation of nitric
modification. Herein, molecular hybridization strategy which is
a common method in drug design [18] was applied to improve the
potency of chalcones and retain their multi-cardiovascular
protective mechanisms. NO donors [19,20] or 1,4-dihydropyridyl
(DHP) moieties [21,22] which are both important pharmacophores
for vasodilators were introduced into the hybrid molecules.
Different NO donors (nitrate or furoxan) were chosen to modulate
the NO release from the final products, while 1,4-DHP moiety was
2
þ
introduced to regulate the Ca entry blockage. Consequently, nine
hybrid chalcone derivatives (Fig. 1) were synthesized and tested for
their vasorelaxant activities through evaluation in aortic rings.
oxide (NO) release, Ca2 entry and inhibition of PKC [10e13].
Superior to traditional vasodilators, the additional antioxidant
properties of flavonoids were also demonstrated to be beneficial in
treatment of cardiovascular diseases [14]. All these facts prompt the
strategies to develop novel and more potent flavonoids as vaso-
relaxant agents.
Previously, we have described the synthesis and biological
evaluation of flavonoid derivatives as vasodilators [15e17]. Struc-
ture-activities relationship (SAR) studies indicated that chalcones
þ
2
2
2
. Results and discussion
.1. Chemistry
.1.1. Synthesis of the hybrid chalcone derivatives conjugated with
NO donors 7b, 8aec and 9a
The synthetic routes of hybrid chalcone derivatives conjugated
with NO donors 7b, 8aec and 9a are summarized in Scheme 1. 4-
Formyl-3-methyl-furoxan 4a was obtained using crotonaldehyde 3
as the starting material according to the literature [23]. Compound
4
a was transformed to its isomer 4b by heating over 98 h in boiling
toluene in moderate yield. Reduction of compounds 4a,b using
NaBH in dioxane, following by treatment with thionyl chloride
*
Corresponding author. Tel./fax: þ86 571 88208460.
E-mail address: huyz@zju.edu.cn (Y. Hu).
4
0223-5234/$ e see front matter Ó 2010 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2010.05.054

X. Dong et al. / European Journal of Medicinal Chemistry 45 (2010) 3986e3992
3987
Fig. 1. Hybrid chalcone derivatives 7b, 8aec, 9a and 15aed.
afforded (chloromethyl)furoxans 5a,b, respectively. Nucleophilic
substitution of chalcone 1a (or 1b) with (chloromethyl)furoxans 5a,
b (or 1,2-dibromoethane) in the presence of potassium carbonate in
chloromethyl-3-methyl-furoxan in the presence of potassium
carbonate provided compound 9a.
acetone resulted in compounds 6aed. Treatment of 6a,b and AgNO
3
2.1.2. Synthesis of the hybrid chalcone derivatives conjugated with
1,4-DHP moiety
The synthesis of the target compounds 15aed is outlined in
Scheme 2. Treatment of m-nitrobenzaldehyde, methyl acetoacetate
in acetonitrile yielded nitrates 7a,b. Deprotection of the phenol
function of 7a and 6c,d in methanol/3N aq. HCl (5:1, v/v) under
reflux afforded compounds 8aec. Etherization of nitrate 8a using 4-
Scheme 1. Synthesis of hybrid chalcone derivatives 7b, 8aec and 9a. Reagents and conditions: (a) acetic acid, NaNO
2
4
, ice bath; (b) toluene, reflux; (c) NaBH , dioxane, r.t.; (d) SOCl
2
,
ꢀ
0
2 3 3 2 3
C to r.t.; (e) K CO , acetone, reflux; (f) AgNO , acetonitrile, r.t.; (g) 3 N HCl, methanol, reflux; (h) K CO , acetone, 5a, reflux.

3988
X. Dong et al. / European Journal of Medicinal Chemistry 45 (2010) 3986e3992
ꢀ
ꢀ
Scheme 2. Synthesis of hybrid chalcone derivatives 15aed. Reagents and conditions: (a) NH
d) K CO , acetone, reflux; (e) 3 N HCl, methanol, reflux.
4
OAc, microwave, 120 C; (b) NaOH, Methanol/H
2
O (6:1, v/v), 70 C; (c) K
2
3
CO , 2, reflux;
(
2
3
and ammonium acetate under microwave irradiation afforded 1,4-
dihydropyridine 10. Selective hydrolysis of 10 in the presence of
sodium hydroxide gave key intermediate monoacid 11 [24,25],
which was then reacted with dibromoethane to yield compound
in compounds 8c (EC50 ¼ 8.9
m
M, Emax ¼ 100.1%). Besides,
increasing the number of nitrates linked to chalcone led to the
enhanced vasorelaxant activities. For example, both of compound
7b bearing three nitrates (EC50 ¼ 8.4
compound 9a bearing nitrate and furoxane (EC50 ¼ 9.4
max ¼ 96.0%) exhibited more potent activities than compound 8a
mM, Emax ¼ 104.0%) and
12. Condensation of 12 with chalcones 13aed in the presence of
mM,
potassium carbonate in acetone under reflux afforded the hybrid
derivatives 14aed. Deprotection of the phenol function of 14aed
was carried out by reaction with methanol/3N aq. HCl (5:1, v/v)
under reflux to yield desired compounds 15aed in good yield.
E
(EC50 ¼ 13.4 M, Emax ¼ 102.9%).
m
SAR of Chalcone hybrid derivatives conjugated to 1,4-DHP moiety:
Comparing the vasorelaxant activities of hybrid compounds 15aed
and 16 indicated that the introduction of 1,4-DHP moiety signifi-
cantly enhanced the biological activities. For instance, the vaso-
relaxant activity of compound 15c was increased by approximately
2.2. Vasorelaxation activity and SAR
6
.7-fold comparing with 16. Insight into the observed effects of
Vasorelaxation activities of hybrid chalcone derivatives 7b,
aec, 9a and 15aed were evaluated in aortic rings with endothe-
different substituent on chalcones revealed that the introduction of
a para-hydroxyl group to the B-ring of chalcone showed optimal
effect (ie. 15c: EC50 ¼ 2.9
hydroxyl groups at the C-5 position of A-ring of chalcone didn’t
exhibit enhanced activity (ie.15a: EC50 ¼ 9.3 M, Emax ¼ 98.6%; 15d:
EC50 ¼ 9.7 M, Emax ¼ 102.9%).
8
lium pre-contracted with 1
mM phenylephrine (PE). Quercetin,
mM, Emax ¼ 104.0%), while introduction of
a well known vasodilator, was used as the positive control. The
results were summarized in Table 1. All tested compounds
promoted relaxation in a dose-dependent manner, and their
m
m
maximal effects were observed at 300 mM as shown in Fig. 2.
SAR of Chalcone hybrid derivatives conjugated with NO donors:
Obviously, the vasorelaxant activities of hybrid chalcone derivatives
3. Conclusion
7
b, 8aec and 9a bearing NO donor were increased when compared
0
0
with that of 2,4,6-trihydroxy-3 ,4 -methylenedioxyl-chalcone 16,
A series of novel hybrid chalcones derivatives conjugated with
NO donors or 1,4-DHP moiety were designed and synthesized
based on the hybrid strategy. Biological activity evaluation showed
most target compounds exhibit significantly improved vaso-
relaxant activities. The preliminary structureeactivity relationships
studies revealed that 4-methyl-3-methylene-furoxan moiety was
the preferable fragment when linked with chalcones, and that the
para-hydroxyl group on B-ring of chalcone was the most optimal
when conjugated with 1,4-DHP moieties. Consequently, compound
a good vasodilator (EC50 ¼ 22.5 M, Emax ¼ 104.1%) reported in our
m
previous studies [15]. Judged by the activities of compounds 8aec,
the 4-methyl-3-methylene-furoxan moiety showed the most
favorable effect on enhancing vasorelaxant activities as exemplified
Table 1
Vasorelaxant activities of hybrid chalcone derivatives 7b, 8aec, 9a and 15aed.
Compd.
EC50
(m
M)
Emax (%)
8c and 15c would be promising structural templates for the
7
8
8
8
9
1
1
1
1
1
b
a
b
c
8.4
104.0 ꢁ 3.6
102.9 ꢁ 5.9
95.7 ꢁ 6.2
development of novel vasorelaxant agents.
13.4
12.1
8.9
9.4
9.3
6.4
2.9
9.7
22.5
244.0
100.1 ꢁ 4.2
96.0 ꢁ 6.7
4. Experiments
a
5a
5b
5c
5d
98.6 ꢁ 6.9
4.1. Chemistry
101.9 ꢁ 7.7
104.0 ꢁ 12.3
102.9 ꢁ 9.1
104.1 ꢁ 17.4
91.3 ꢁ 13.2
Reagents and solvents were purchased from commercial sour-
a
6
ces. Melting-points were obtained on a B-540 Büchi melting-point
apparatus and are uncorrected. Intermediates and target
a
Quercetin
a
1
Data available in our previous studies [15].
compounds were characterized based on H NMR (Brüker AM500)

X. Dong et al. / European Journal of Medicinal Chemistry 45 (2010) 3986e3992
3989
Fig. 2. Effects of hybrid chalcone derivatives on relaxation in aortic rings with endothelium pre-contracted with 1
a were added cumulatively to achieve the appropriate concentrations; (b) The tested compounds 15aed were added cumulatively to achieve the appropriate concentrations.
Results were expressed as means ꢁ standard error of mean in terms of percentage relaxation of the contraction to PE (n ¼ 3e4).
mM phenylephrine (PE). (a) The tested compounds 7b, 8aec and
9
and 13C NMR (Brüker AM125). Mass spectra were recorded on an
Esquire-LC-00075 spectrometer. High resolution mass spectral
J ¼ 6.5 Hz), 5.19 (s, 2H), 5.22 (s, 2H), 6.00 (s, 2H), 6.34 (d, 1H,
J ¼ 2.0 Hz), 6.55 (d, 1H, J ¼ 2.0 Hz), 6.79 (d, 1H, J ¼ 8.5 Hz), 6.83 (d,
1H, J ¼ 16.5 Hz), 7.00 (dd, 1H, J ¼ 1.5, 8.5 Hz), 7.07 (d, 1H, J ¼ 1.5 Hz),
7.28 (d, 1H, J ¼ 16.5 Hz). ESI-MS: m/z [M þ H]þ 495.
(
HRMS) analyses were recorded on Brüker DALTONICS APEX III
mass spectrometer using electron spray ionization. Chalcone 1a,
b [15], furoxane 4a,b [23] and 1,4-DHPs 11 [24,25] were synthesized
according to the literature.
4.2.5. 3-(Benzo[d][1,3]dioxol-5-yl)-1-(2,4,6-tris(2-bromoethoxy)
phenyl)prop-2-en-1-one (6b)
Reagent: compound 1b (300 mg, 1.00 mmol), potassium
4
.2. General procedure for the synthesis of furoxane 5a,b
carbonate (276 mg, 2.00 mmol), 1,2-dibromoethane (4.83 g,
1
2
d
5.7 mmol); yellow syrup (416 mg, 67%). H NMR (CDCl
3
, 500 MHz,
NaBH
4
(300 mg, 7.9 mmol) was added to a cold solution of 4a (or
): 3.52 (t, 4H, J ¼ 6.5 Hz), 3.66 (t, 2H, J ¼ 6.5 Hz), 4.26 (t, 4H,
J ¼ 6.5 Hz), 4.30 (t, 2H, J ¼ 6.5 Hz), 6.00 (s, 2H), 6.18 (s, 2H), 6.79 (d,
H, J ¼ 7.5 Hz), 6.82 (d, 1H, J ¼ 16.5 Hz), 7.00 (dd, 1H, J ¼ 1.5, 7.5 Hz),
4
b) (500 mg, 3.84 mmol) in dioxane (5 mL). The reaction mixture
was stirred for 12 h at room temperature. The residue obtained by
concentration of the solution was treated with water and extracted
with ethyl acetate (15 mL ꢂ 3). The organic phase was evaporated in
vacuo to give pale yellow oil, to which thionyl chloride (1.67 g,
1
7
[
.06 (d, 1H, J ¼ 1.5 Hz), 7.26 (d, 1H, J ¼ 16.5 Hz). ESI-MS: m/z
M þ H]þ 619.
1
4.1 mmol) was added dropwise in ice-water bath. The reaction
4
(
.2.6. 4-((2-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)-3,5-bis
methoxymethoxy)phenoxy)methyl)-3-methyl-1,2,5-oxadiazole 2-
oxide (6c)
Reagent: compound 1a (200 mg, 0.52 mmol), potassium
carbonate (142 mg,1.04 mmol) and 4a (80.8 mg, 0.55 mmol); yellow
mixture was kept under stirring at room temperature for 12 h.
Upon finishing of the reaction, water (50 mL) was added, and the
mixture was extracted with ethyl acetate (20 mL ꢂ 3). Evaporating
of the dried combined organic layers afforded the crude product as
an oily residue which was purified by silica gel column chroma-
tography using gradient elution with petroleum ethereethyl
acetate (10:1e6:1).
1
syrup (206 mg, 80%). H NMR (CDCl
3
, 500 MHz,
d): 2.31 (s, 3H), 3.37
(s, 3H), 3.51 (s, 3H), 5.00 (s, 2H), 5.10 (s, 2H), 5.20 (s, 2H), 6.01 (s, 2H),
6
6
.43 (d,1H, J ¼ 2.0 Hz), 6.57 (d,1H, J ¼ 2.0 Hz), 6.75 (d,1H, J ¼ 16.5 Hz),
.80 (d, 1H, J ¼ 8.0 Hz), 6.96 (dd, 1H, J ¼ 1.5, 8.0 Hz), 7.01 (d, 1H,
4.2.1. 3-Methyl-4-chloromethyl-furoxane 5a
þ
J ¼ 1.5 Hz), 7.21 (d, 1H, J ¼ 16.5 Hz). ESI-MS: m/z [M þ H] 501.
1
Pale yellow oil (45%); H NMR (CDCl
); 4.39 (s, 2H, CH ).
3
, 400 MHz,
d
d
): 2.22 (s, 3H,
): 2.45 (s, 3H,
CH
3
2
4
(
.2.7. 3-((2-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)-3,5-bis
methoxymethoxy)phenoxy)methyl)-4-methyl-1,2,5-oxadiazole 2-
oxide (6d)
Reagent: compound 1a (200 mg, 0.52 mmol), potassium
carbonate (142 mg, 1.04 mmol) and 4b (81 mg, 0.55 mmol); yellow
4.2.2. 3-Chloromethyl-4-methyl-furoxane 5b
1
Pale yellow oil (38%); H NMR (CDCl
); 4.26 (s, 2H, CH ).
3
, 400 MHz,
CH
3
2
1
syrup (196 mg, 76%). H NMR (CDCl
3
, 500 MHz,
d): 2.12 (s, 3H), 3.38
4.2.3. General procedure for the synthesis of compounds 6aed
(s, 3H), 3.51 (s, 3H), 5.10 (s, 2H), 5.11 (s, 2H), 5.20 (s, 2H), 6.01 (s, 2H),
A solution of compounds 1, potassium carbonate and 1,2-
6
6
.48 (d,1H, J ¼ 2.0 Hz), 6.57 (d,1H, J ¼ 2.0 Hz), 6.76 (d,1H, J ¼ 16.5 Hz),
dibromoethane 2 (or 5a,b) in acetone (20 mL) was refluxed for
e10 h. After cooled to room temperature, the mixture was filtered
.80 (d, 1H, J ¼ 8.0 Hz), 6.97 (dd, 1H, J ¼ 1.5, 8.0 Hz), 7.03 (d, 1H,
2
þ
J ¼ 1.5 Hz), 7.21 (d, 1H, J ¼ 16.5 Hz). ESI-MS: m/z [M þ H] 501.
and the solution was concentrated in vacuo. The residue was
purified by silica gel column chromatography using gradient
elution with petroleum ethereethyl acetate (10:1e4:1) to give
expected product.
4.3. General procedure for the synthesis of nitrate 7a,b
To a stirred solution of 6a,b in acetonitrile (5 mL), silver nitrate
was added. The reaction mixture was stirred in the dark for
50e70 h. The reaction mixture was then filtered, and the filtrate
was evaporated under reduced pressure to afford a dark-yellow
solid which was purified by silica gel column chromatography
using gradient elution with petroleum ethereethyl acetate
(5:1e2:1) to give expected product.
4
.2.4. 3-(Benzo[d][1,3]dioxol-5-yl)-1-(2-(2-bromoethoxy)-4,6-bis
(
methoxymethoxy)phenyl)prop-2-en-1-one (6a)
Reagent: compound 1a (300 mg, 1.29 mmol), potassium
carbonate (356 mg, 2.58 mmol), 1,2-dibromoethane (4.83 g,
2
1
5.7 mmol); yellow syrup (359 mg, 56%). H NMR (CDCl
3
, 500 MHz,
d): 3.40 (s, 3H), 3.51 (s, 3H), 3.52 (t, 2H, J ¼ 6.5 Hz), 4.26 (t, 2H,

3990
X. Dong et al. / European Journal of Medicinal Chemistry 45 (2010) 3986e3992
4
.3.1. 2-(2-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)-3,5-bis
1H, J ¼ 2.0 Hz), 6.92 (d, 1H, J ¼ 8.0 Hz), 7.15 (dd, 1H, J ¼ 1.5, 8.0 Hz),
7.17 (d, 1H, J ¼ 1.5 Hz), 7.68 (d, 1H, J ¼ 15.0 Hz), 7.72 (d, 1H,
(
methoxymethoxy)phenoxy)ethyl nitrate (7a)
13
Reagent: compound 6a (300 mg, 0.61 mmol), silver nitrate
J ¼ 15.0 Hz), 9.50 (brs, 1H, OH), 14.10 (s, 1H, OH). C NMR (Acetone-
1
(
277 mg, 1.633 mmol); yellow syrup (225 mg, 78%). H NMR (CDCl
3
,
6
d , 125 MHz, d): 193.0, 165.9, 163.2, 161.2, 154.8, 150.7, 148.3, 144.4,
5
00 MHz,
d
): 3.40 (s, 3H), 3.51 (s, 3H), 4.23 (t, 2H, J ¼ 6.5 Hz), 4.71 (t,
130.5, 124.8, 123.6, 112.9, 109.1, 108.9, 108.0, 102.1, 96.6, 93.6, 58.6,
þ
2
H, J ¼ 6.5 Hz), 5.20 (s, 2H), 5.23 (s, 2H), 6.00 (s, 2H), 6.34 (d, 1H,
8.2. HRMS(ESI): Found 413.0918 ([M þ H] ), calcd for C20
16 2 8
H N O :
J ¼ 2.0 Hz), 6.55 (d, 1H, J ¼ 2.0 Hz), 6.80 (d, 1H, J ¼ 8.5 Hz), 6.83 (d,
412.0907.
1
7
H, J ¼ 16.5 Hz), 7.01 (dd, 1H, J ¼ 1.5, 8.5 Hz), 7.06 (d, 1H, J ¼ 1.5 Hz),
.29 (d, 1H, J ¼ 16.5 Hz). ESI-MS: m/z [M þ H]þ 478.
4.4.4. 4-((4-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)-3-hydroxy-5-(2-
nitrooxy)ethoxy)phenoxy)methyl)-3-methyl-1,2,5-oxadiazole 2-
oxide (9a)
(
0 00
.3.2. 2,2 ,2 -(2-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)benzene-
4
1
,3,5-triyl)tris(oxy)tris(ethane-2,1-diyl) trinitrate (7b)
Reagent: compound 6b (400 mg, 0.64 mmol), silver nitrate
A solution of compound 8a (50 mg, 0.13 mmol), potassium
carbonate (35 mg, 0.25 mmol) and 5a in acetone (3 mL) was
refluxed for 10 h. The mixture was filtered and the solution was
concentrated in vacuo. The residue was purified by chromatography
on silica gel with petroleum ethereethyl acetate (3:1) to give
ꢀ
(
277 mg, 1.633 mmol); yellow solid (237 mg, 78%), mp 101e103 C.
1
H NMR (CDCl
J ¼ 6.5 Hz), 4.75 (t, 4H, J ¼ 6.5 Hz), 4.88 (t, 2H, J ¼ 6.5 Hz), 6.05 (s,
H), 6.23 (s, 2H), 6.82 (d, 1H, J ¼ 16.5 Hz), 6.85 (d, 1H, J ¼ 7.5 Hz),
.04 (dd, 1H, J ¼ 1.5, 7.5 Hz), 7.11 (d, 1H, J ¼ 1.5 Hz), 7.27 (d, 1H,
3
, 500 MHz,
d): 4.27 (t, 4H, J ¼ 6.5 Hz), 4.33 (t, 2H,
ꢀ
1
2
7
product 9a as pale yellow solid (36 mg, 56%), mp 95e97 C. H NMR
(CDCl , 400 MHz,
3
d
): 2.14 (s, 3H), 4.55 (t, 2H, J ¼ 6.5 Hz), 5.13 (t, 2H,
13
J ¼ 16.5 Hz). C NMR (Acetone-d
6
, 125 MHz,
d): 191.2, 164.0, 162.6,
J ¼ 6.5 Hz), 5.46 (s, 2H), 6.05 (d, 1H, J ¼ 2.0 Hz), 6.10 (s, 2H), 6.21 (d,
1H, J ¼ 2.0 Hz), 6.84 (d, 1H, J ¼ 8.0 Hz), 7.06 (dd, 1H, J ¼ 1.5, 8.0 Hz),
7.11 (d, 1H, J ¼ 1.5 Hz), 7.65 (d, 1H, J ¼ 15.6 Hz), 7.81 (d, 1H,
1
6
C
51.1, 146.3, 144.4, 132.3, 121.8, 119.6, 115.4, 108.2, 107.8, 102.1, 94.3,
þ
6.6, 62.9, 58.2. HRMS(ESI): Found 568.0980 ([M þ H] ), calcd for
15: 567.0973.
1
3
H
22 21
N
3
O
J ¼ 15.6 Hz), 13.95 (s, 1H, OH). C NMR (Acetone-d
6
, 125 MHz, d):
1
93.4, 165.6, 164.3, 161.8, 157.5, 150.1, 148.3, 143.8, 130.3, 124.9,
4
.4. General procedure for the synthesis of compounds 8aec
123.3, 113.5, 109.7, 109.2, 108.7, 102.3, 96.2, 93.5, 71.8, 64.9, 60.6, 8.2.
þ
HRMS(ESI): Found 502.1014 ([M þ H] ), calcd for C22
19 3 11
H N O :
To a solution of 6c,d (or 7a) in methanol (2 mL), 3 N aq. HCl
501.1020.
(
0.4 mL) was added. The resulting mixture was refluxed for 45 min,
then poured into cold water and extracted with ethyl acetate
5 mL ꢂ 3 Times). The organic phase was washed with brine and
then dried over anhydrous Na SO . After removal of the solvent, the
4.5. 3-(2-Bromoethyl)-5-methyl-2,6-dimethyl-4-(3-nitrophenyl)-
(
1,4-dihydropyridine-3,5-dicarboxylate (12)
2
4
residue was purified by chromatography on silica gel using gradient
elution with petroleum ethereethyl acetate (8:1e3:1) to give
expected product.
A solution of compound 11 (1.2 g, 3.61 mmol), potassium
carbonate (0.99 g, 7.22 mmol) and 1,2-dibromoethane 2 (13.5 g,
2.2 mmol) in acetone (20 mL) was refluxed for 5 h. The mixture
7
was filtered and the solution was concentrated in vacuo. The
residue was purified by silica gel column chromatography using
4
.4.1. 2-(2-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)-3,5-
dihydroxyphenoxy)ethyl nitrate (8a)
gradient elution with petroleum ethereethyl acetate (4:1e2:1) to
Reagent: compound 7a (100 mg, 0.21 mmol); pale yellow solid
1
give title product as yellow syrup (1.05 g, 66%). H NMR (CDCl
3
,
ꢀ
1
(
65 mg, 80%), mp 193e194 C. H NMR (Acetone-d , 500 MHz, d):
6
500 MHz, d): 2.37 (s, 3H), 2.39 (s, 3H), 3.46 (m, 2H), 3.65 (s, 3H), 4.37
4
6
1
.57 (t, 2H, J ¼ 6.5 Hz), 5.12 (t, 2H, J ¼ 6.5 Hz), 6.07 (d,1H, J ¼ 2.0 Hz),
(
m, 2H), 5.12 (s, 1H), 5.83 (s, 1H), 7.38 (t, 1H, J ¼ 8.0 Hz), 7.67 (dd, 1H,
.12 (s, 2H), 6.13 (d, 1H, J ¼ 2.0 Hz), 6.93 (d, 1H, J ¼ 8.0 Hz), 7.29 (dd,
H, J ¼ 1.5, 8.0 Hz), 7.33 (d, 1H, J ¼ 1.5 Hz), 7.76 (d, 1H, J ¼ 16.5 Hz),
J ¼ 2.0, 8.0 Hz), 8.00 (dd, 1H, J ¼ 2.0, 8.0 Hz), 8.11 (d, 1H, J ¼ 8.0 Hz).
ESI-MS: m/z [M þ H]þ 439.
13
7
.90 (d, 1H, J ¼ 16.5 Hz), 9.54 (s, 1H, OH), 14.17 (s, 1H, OH). C NMR
(
Acetone-d
6
, 125 MHz,
d): 193.4, 165.9, 162.3, 161.1, 150.0, 147.3,
1
6
3
44.4, 130.3, 125.8, 123.6, 109.2, 108.4, 107.6, 102.1, 96.6, 93.3, 70.1,
4.6. General procedure for the synthesis of compounds 14a,d
þ
4.8. HRMS(ESI): Found 390.0755 ([M þ H] ), calcd for C18
9
H15NO :
89.0747.
A solution of compound 12, potassium carbonate and appro-
priate chalcone 13a,d in acetone (5 mL) was refluxed for 4e12 h.
The mixture was filtered and the solution was concentrated in
vacuo. The residue was purified by silica gel column chromatog-
raphy using gradient elution with petroleum ethereethyl acetate
(4:1e1:1) to give expected product.
4.4.2. 4-((2-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)-3,5-
dihydroxyphenoxy)methyl)-3-methyl-1,2,5-oxadiazole 2-oxide (8b)
Reagent: compound 6c (150 mg, 0.3 mmol); pale yellow solid
ꢀ
1
(
2
1
68 mg, 55%), mp 220e222 C. H NMR (Acetone-d
6
, 500 MHz,
.13 (s, 3H), 5.44 (s, 2H), 6.00 (s, 2H), 6.01 (d, 1H, J ¼ 2.0 Hz), 6.26 (d,
H, J ¼ 2.0 Hz), 6.88 (d, 1H, J ¼ 8.0 Hz), 7.08 (dd, 1H, J ¼ 1.5, 8.0 Hz),
d):
4.6.1. 3-(2-(2-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)-3,5-bis
(methoxymethoxy)phenoxy)ethyl)-5-methyl-2,6-dimethyl-4-(3-
nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate (14a)
Reagent: compound 12 (200 mg, 0.45 mmol), potassium
carbonate (124 mg, 0.9 mmol) and 13a (152 mg, 0.45 mmol); pale
7
.09 (d, 1H, J ¼ 1.5 Hz), 7.63 (d, 1H, J ¼ 15.0 Hz), 7.78 (d, 1H,
J ¼ 15.0 Hz), 9.52 (brs, 1H, OH), 14.13 (s, 1H, OH). ESI-MS: m/z
þ
13
[
1
M þ H] 413. C NMR (Acetone-d
6
, 125 MHz, d): 192.8, 166.1, 163.2,
60.8, 155.4, 151.2, 147.2, 144.3, 130.8, 124.9, 124.1, 112.3, 109.2,
1
1
108.8, 107.2, 102.3, 96.5, 93.6, 60.5, 7.8. HRMS(ESI): Found 413.0920
yellow syrup (238 mg, 70%). H NMR (CDCl
(CDCl , 400 MHz, ): 2.22 (s, 3H), 2.32 (s, 3H), 3.39 (s, 3H), 3.48 (m,
H), 3.60 (s, 3H), 4.28 (m, 2H), 5.01 (s, 1H), 5.12 (s, 2H), 5.20 (s, 2H),
3
, 500 MHz, d): H NMR
þ
(
[M þ H] ), calcd for C20
H
16
N
2
O
8
: 412.0907.
3
d
2
4
.4.3. 3-((2-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)-3,5-
5.60 (s, 1H), 6.05 (s, 2H), 6.29 (d, 1H, J ¼ 2.0 Hz), 6.52 (d, 1H,
J ¼ 2.0 Hz), 6.82 (d, 1H, J ¼ 8.5 Hz), 6.92 (d, 1H, J ¼ 16.0 Hz), 7.05 (dd,
1H, J ¼ 1.5, 8.5 Hz), 7.11 (d, 1H, J ¼ 1.5 Hz), 7.28 (t, 2H, J ¼ 8.0 Hz),
7.58 (d, 1H, J ¼ 16.0 Hz), 7.61 (d, 1H, J ¼ 8.0 Hz), 7.96 (dd, 1H, J ¼ 1.5,
8.0 Hz), 8.01 (d, 1H, J ¼ 1.5 Hz). ESI-MS: m/z [M þ H]þ 746.
dihydroxyphenoxy)methyl)-4-methyl-1,2,5-oxadiazole 2-oxide (8c)
Reagent: compound 6c (150 mg, 0.3 mmol); pale yellow solid
ꢀ
1
(
78 mg, 63%), mp 198e201 C. H NMR (Acetone-d
6
, 500 MHz, d):
2
.47 (s, 3H), 5.33 (s, 2H), 6.11 (d, 1H, J ¼ 2.0 Hz), 6.12 (s, 2H), 6.22 (d,

X. Dong et al. / European Journal of Medicinal Chemistry 45 (2010) 3986e3992
3991
4
.6.2. 3-(2-(2-(3-Bromophenyl)acryloyl)-(3,5-bis
192.3, 169.6, 169.5, 167.5, 163.93, 162.32, 151.2, 150.4, 149.2, 147.8,
145.6, 144.7, 138.5, 132.3, 130.3, 129.3, 127.9, 124.7, 124.6, 122.6,
(
methoxymethoxy)phenoxy)ethyl)-5-methyl-2,6-dimethyl-4-(3-
nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate (14b)
109.2, 108.9, 107.2, 103.8, 102.1, 96.6, 94.2, 93.8, 67.9, 61.3, 52.0, 35.6,
þ
Reagent: compound 12 (200 mg, 0.45 mmol), potassium
16.3. HRMS(ESI): Found 659.1687 ([M
34 30 2
C H N O12: 658.1799.
þ
H] ), calcd for
carbonate (124 mg, 0.9 mmol)and 13b (193 mg, 0.45 mmol); pale
1
yellow syrup (203 mg, 57%). H NMR (CDCl
3
, 400 MHz,
d
): 2.24 (s,
3
5
H), 2.34 (s, 3H), 3.40 (s, 3H), 3.51 (m, 2H), 3.62 (s, 3H), 4.26 (m, 2H),
.00 (s, 1H), 5.11 (s, 2H), 5.18 (s, 2H), 5.64 (s, 1H), 6.30 (d, 1H,
4.7.2. 3-(2-(2-(3-(3-Bromophenyl)acryloyl)-3,5-
dihydroxyphenoxy)ethyl)5-methyl,6-dimethyl -4-(3-nitrophenyl)-
1,4-dihydropyridine-3,5-dicarboxylate (15b)
J ¼ 2.0 Hz), 6.52 (d, 1H, J ¼ 2.0 Hz), 6.90 (d, 1H, J ¼ 16.0 Hz), 7.22 (t,
2
1
H, J ¼ 7.5 Hz), 7.27 (t, 2H, J ¼ 8.0 Hz), 7.40 (d,1H, J ¼ 7.5 Hz), 7.46 (d,
H, J ¼ 7.5 Hz), 7.55 (d, 1H, J ¼ 16.0 Hz), 7.60 (d, 1H, J ¼ 8.0 Hz), 7.61
Reagent: compound 14b (100 mg, 0.13 mmol), pale yellow solid
ꢀ
1
6
(62 mg, 70%); mp 148e150 C. H NMR (Acetone-d , 500 MHz, d):
(
s, 1H), 7.92 (dd, 1H, J ¼ 1.5, 8.0 Hz), 8.03 (d, 1H, J ¼ 1.5 Hz). ESI-MS:
2.31 (s, 6H), 3.52 (s, 3H), 4.25 (m, 1H), 4.45 (m, 1H), 4.49 (m, 1H),
þ
m/z [M þ H] 781.
4.63 (m, 1H), 5.00 (s, 1H), 5.65 (s, 1H), 6.10 (d, 1H, J ¼ 2.5 Hz), 6.11 (d,
1
H, J ¼ 2.5 Hz), 7.26 (t, 1H, J ¼ 8.0 Hz), 7.29 (t, 1H, J ¼ 8.0 Hz), 7.52
4.6.3. 3-(2-(3,5-Bis(methoxymethoxy)-2-(3-(4-(methoxymethoxy)
(dd, 1H, J ¼ 1.0, 8.0 Hz), 7.56 (t, 2H, J ¼ 8.0 Hz), 7.63 (d, 1H,
J ¼ 16.5 Hz), 7.77 (d, J ¼ 1.0 Hz), 7.87 (dd, 1H, J ¼ 1.0, 8.0 Hz), 8.00 (d,
1H, J ¼ 1.0 Hz), 8.13 (d, 1H, J ¼ 16.5 Hz), 9.52 (s, 1H, OH), 14.40 (s, 1H,
phenyl)acryloyl)phenoxy)ethyl) 5-methyl-2,6-dimethyl-4-(3-
nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate (14c)
Reagent: compound 12 (200 mg, 0.45 mmol), potassium
13
6
OH). C NMR (Acetone-d , 125 MHz, d): 193.4, 169.8, 169.5, 165.6,
carbonate (124 mg, 0.9 mmol) and 13c (184 mg, 0.45 mmol); pale
163.4, 162.1,149.2, 145.6, 144.9,144.7, 138.9, 138.1,136.5, 132.3, 131.1,
130.7, 129.5, 128.1, 127.2, 126.1, 125.1122.2, 109.2, 106.8, 96.6, 93.8,
1
yellow syrup (219 mg, 63%). H NMR (CDCl
3
, 400 MHz,
d): 2.19 (s,
3
3
H), 2.33 (s, 3H), 3.39 (s, 3H), 3.46 (m, 2H), 3.47 (s, 3H), 3.51 (s, 3H),
.62 (s, 3H), 4.28 (m, 2H), 5.04 (s, 1H), 5.11 (s, 2H), 5.18 (s, 2H), 5.20
95.4, 94.2, 67.5, 61.5, 52.1, 34.9, 16.8. HRMS(ESI): Found 693.1017
þ
2
([M þ H] ), calcd for C33H29BrN O10: 692.1006.
(
6
s, 2H), 5.63 (s, 1H), 6.30 (d, 1H, J ¼ 2.0 Hz), 6.53 (d, 1H, J ¼ 2.0 Hz),
.81 (d, 1H, J ¼ 16.0 Hz), 7.00 (d, 2H, J ¼ 8.5 Hz), 7.30 (t, 1H,
4.7.3. 3-(2-(3,5-Dihydroxy-2-(3-(4-hydroxyphenyl)acryloyl)
phenoxy)ethyl)-5-methyl-2,6-dimethyl-4-(3-nitrophenyl)-1,4-
dihydropyridine-3,5-dicarboxylate (15c)
J ¼ 8.0 Hz), 7.42 (d, 2H, J ¼ 8.5 Hz), 7.61 (d, 1H, J ¼ 16.0 Hz), 7.94 (dd,
1
H, J ¼ 2.0, 8.0 Hz), 8.04 (d, 1H, J ¼ 2.0 Hz). ESI-MS: m/z [M þ H]þ
763.
Reagent: compound 14c (100 mg, 0.13 mmol), pale yellow solid
ꢀ
1
(
48 mg, 58%); mp 172e174 C. H NMR (Acetone-d , 500 MHz, d):
6
4
.6.4. 3-(2-(2-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)-5-
2.33 (s, 6H), 3.56 (s, 3H), 4.24 (m, 1H), 4.46 (m, 2H), 4.50 (m, 1H),
4.65 (m, 1H), 5.01 (s, 1H), 5.67 (s, 1H), 6.12 (d, 1H, J ¼ 2.0 Hz), 6.13 (d,
1H, J ¼ 2.0 Hz), 6.82 (d, 2H, J ¼ 8.5 Hz), 7.56 (t, 2H, J ¼ 8.5 Hz), 7.59
(d, 2H J ¼ 8.0 Hz), 7.63 (d, 1H, J ¼ 16.5 Hz), 7.87 (dd, 1H, J ¼ 2.0,
(
methoxymethoxy)phenoxy)ethyl) 5-methyl 2,6-dimethyl-4-(3-
nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate (14d)
Reagent: compound 12 (200 mg, 0.45 mmol), potassium
carbonate (124 mg, 0.9 mmol)and 13d (149 mg, 0.45 mmol); pale
yellow syrup (219 mg, 70%). H NMR (CDCl
8.5 Hz), 8.00 (d, 1H, J ¼ 2.0 Hz), 8.07 (d, 1H, J ¼ 16.5 Hz), 9.05 (s, 1H,
1
13
3
, 500 MHz,
d
): 2.21 (s,
OH), 9.55 (s, 1H, OH), 14.33 (s, 1H, OH). C NMR (Acetone-d
6
,
3
2
H), 2.34 (s, 3H), 3.44 (s, 3H), 3.44 (m, 2H), 3.63 (s, 3H), 4.23 (m,
H), 5.01 (s, 1H), 5.12 (s, 2H), 5.62 (s, 1H), 6.02 (s, 2H), 6.65 (dd, 1H,
125 MHz, d): 194.2, 169.5, 169.3, 164.3, 163.4, 162.1, 160.1, 150.2,
148.1, 145.6, 144.7, 138.7, 136.6, 129.7, 129.5, 127.4, 126.8, 125.3,
J ¼ 2.0, 8.0 Hz), 6.73 (d, 1H, J ¼ 2.0 Hz), 7.02 (d, 1H, J ¼ 16.0 Hz), 6.85
124.6, 116.4, 108.9, 96.3, 94.0, 93.8, 93.5, 67.7, 62.3, 52.0, 35.8, 16.7.
þ
(
d, 1H, J ¼ 8.4 Hz), 7.14 (dd, 1H, J ¼ 2.0, 8.4 Hz), 7.20 (d, 1H,
HRMS(ESI): Found 631.1859 ([M þ H] ), calcd for C33
30 2 11
H N O :
J ¼ 2.0 Hz), 7.35 (t, 2H, J ¼ 8.0 Hz), 7.55 (d, 1H, J ¼ 16.0 Hz), 7.80 (d,
630.1850.
1
H, J ¼ 8.0 Hz), 7.97 (dd, 1H, J ¼ 2.0, 8.0 Hz), 8.03 (d, 1H, J ¼ 2.0 Hz).
þ
ESI-MS: m/z [M þ H] 687.
4.7.4. 3-(2-(2-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)-5-
hydroxyphenoxy)ethyl)-5-methyl-2,6-dimethyl-4-(3-nitrophenyl)-
4.7. General procedure for the synthesis of hybrid compound 15aed
1,4-dihydropyridine-3,5-dicarboxylate (15d)
Reagent: compound 14d (100 mg, 0.15 mmol), pale yellow solid
ꢀ
1
To a solution of 14aed in methanol (2 mL), 3 N HCl aqueous
6
(67 mg, 72%); mp 139e141 C. H NMR (Acetone-d , 500 MHz, d):
(
0.4 mL) was added. The resulting mixture was refluxed for 1.5 h,
then poured into cold water and extracted with ethyl acetate
5 mL ꢂ 3 Times). The organic phase was washed with brine and
then dried over anhydrous Na SO . After removal of the solvent, the
2.29 (s, 3H), 2.30 (s, 3H), 3.49 (s, 3H), 4.27 (m,1H), 4.42 (m, 2H), 4.50
(m, 1H), 5.03 (s, 1H), 5.61 (s, 1H), 6.04 (s, 2H), 6.57 (d, 1H, J ¼ 2.5 Hz),
6.59 (s, 1H), 6.78 (d, 1H, J ¼ 3.0 Hz), 7.06 (d, 1H, J ¼ 3.0 Hz), 7.07 (s,
1H), 7.25 (t, 1H, J ¼ 8.0 Hz), 7.50 (d, 1H, J ¼ 15.5 Hz), 7.58 (d, 1H,
J ¼ 8.0 Hz), 7.65 (d, 1H, J ¼ 15.5 Hz), 7.77 (d, 1H, J ¼ 2.5 Hz), 7.86 (d,
(
2
4
residue was purified by chromatography on silica gel using gradient
elution with petroleum ethereethyl acetate (2:1e1:2) to give
expected product.
13
1H, J ¼ 3.0 Hz), 8.03 (s, 1H), 9.09 (s, 1H, OH). C NMR (Acetone-d
6
,
125 MHz,
d): 191.1, 169.6, 169.4, 162.2, 161.6, 150.8, 148.3, 147.7,
144.9, 144.6, 143.4, 139.3, 136.6, 132.2, 130.7, 129.5, 127.9, 124.5,
4
.7.1. 3-(2-(2-(3-(Benzo[d][1,3]dioxol-5-yl)acryloyl)-3,5-
123.7, 121.9, 119.8, 109.4, 109.2, 108.8, 102.9, 102.4, 96.6, 94.2, 67.7,
þ
dihydroxyphenoxy)ethyl)-5-methyl 2,6-dimethyl-4-(3-
nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate (15a)
Reagent: compound 14a (100 mg, 0.13 mmol), pale yellow solid
62.3, 52.0, 35.8, 16.5. HRMS(ESI): Found 643.1860 ([M þ H] ), calcd
30 2
for C34H N O11: 642.1850.
ꢀ
1
(
2
(
1
57 mg, 65%); mp 162e165 C. H NMR (Acetone-d
.32 (s, 3H), 2.33 (s, 3H), 3.54 (s, 3H), 4.24 (m, 1H), 4.46 (m, 1H), 4.49
m, 1H), 4.62 (m, 1H), 5.02 (s, 1H), 5.64 (s, 1H), 6.08 (s, 2H), 6.12 (d,
H, J ¼ 2.0 Hz), 6.13 (s, 1H, J ¼ 2.0 Hz), 6.95 (d, 1H, J ¼ 8.0 Hz), 7.30
dd, 1H, J ¼ 1.5, 8.0 Hz), 7.33 (d, 1H, J ¼ 1.5 Hz), 7.56 (t, 2H,
6
, 500 MHz,
d
):
4.8. Vasodilatory effect assay
Vascular rings were prepared from the aorta of male Male
SpragueeDawley rats (four to six months old and weighing on
average 250 g), and contraction studies were performed following
the general procedure detailed in the literature [26]. After an
equilibration period of at least 1 h, isometric contractions induced
(
J ¼ 8.0 Hz), 7.74 (d,1H, J ¼ 16.5 Hz), 7.87 (dd,1H, J ¼ 2.0, 8.0 Hz), 7.88
(
9
d, 1H, J ¼ 16.5 Hz), 8.00 (d, 1H, J ¼ 2.0 Hz), 8.10 (d, 1H, J ¼ 16.5 Hz),
13
.54 (s,1H, OH),14.17 (s,1H, OH). C NMR (Acetone-d
6
,125 MHz,
d):
by PE (1 mM) were obtained. When contraction of the tissue in

3992
X. Dong et al. / European Journal of Medicinal Chemistry 45 (2010) 3986e3992
response to this vasoconstrictor agent had stabilized (after about
0 min), cumulatively increasing concentrations of the tested
[6] Z. Nowakowska, Eur. J. Med. Chem. 42 (2007) 125e137.
[
[
[
7] L. Hooper, P. AKroon, E.B. Rimm, J.S. Cohn, I. Harvey, K.A. Cornu, J.J. Ryder, W.
L. Hall, A. Cassidy, Am. J. Clin. Nutr. 88 (2008) 38e50.
8] F. Fusi, S. Saponara, F. Pessina, B. Gorelli, G. Sgaragli, Eur. J. Nutr. 42 (2003)
10e17.
9] J. Duarte, F. Perez-vizcaino, J. Jimenez, J. Tamargo, A. Zarzuelo, Stud. Nat. Prod.
Chem. 25 (2001) 565e605.
2
compounds were added to the bath at 15e20 min intervals (the
time needed to obtain steady-state relaxation). Control tissues
were simultaneously subjected to the same procedures, but omit-
ting the compounds and adding the vehicle. The flavonoids-
induced maximal relaxation (Emax) in aortic rings was calculated as
a percentage of the contraction in response to PE (1 mM). The half
maximum effective concentration (EC50) was defined as the
concentration of the flavonoids that induced 50% of maximum
[
[
[
[
[
[
[
[
10] Y.H. Zhang, Y.S. Park, T.J. Kim, L.H. Fang, H.Y. Ahn, J.T. Hong, Y. Kim, C.K. Lee, Y.
P. Yun, Gen. Pharmacol. 35 (2002) 341e347.
11] S.K. Mishra, S.E. Abbot, A. Choudhury, M. Cheng, N. Khatab, N.J.R. Maycock,
A. Zavery, P.I. Aaronson, Cardiovasc. Res. 46 (2000) 539e546.
12] H.D. Jiang, Q. Xia, X.X. Wang, J.F. Song, I.C. Jiang, Pharmazie 60 (2005)
444e447.
13] Z. Pan, T. Feng, L. Shan, B. Cai, W. Chu, H. Niu, Y. Lu, B. Yang, Phytother. Res. 22
relaxation from the contraction elicited by PE (1
mM) and was
(2008) 1428e1433.
calculated from the concentration response curve by nonlinear
regression (curve fit) using GraphPad Prism (Version 4.0).
14] M. Galisteo, M.F. García-Saura, R. Jiménez, C.I. Villar, A. Zarzuelo, F. Vargas,
J. Duarte, Mol. Cell. Biochem. 259 (2004) 91e99.
15] X. Dong, T. Liu, J. Yan, P. Wu, J. Chen, Y. Hu, Bioorg. Med. Chem. 17 (2009)
716e726.
Acknowledgement
16] X. Dong, L. Qi, C. Jiang, J. Chen, E. Wei, Y. Hu, Bioorg. Med. Chem. Lett. 19
(2009) 3196e3198.
The authors are grateful to support from the Administration of
Traditional Chinese Medicine of Zhejiang Province (NO.
17] X. Dong, J. Chen, C. Jiang, T. Liu, Y. Hu, Arch. Pharm. 342 (2009) 428e432.
[18] C. Viegas-Junior, A. Danuello, V. da Silva Bolzani, E.J. Barreiro, C.A.M. Fraga,
Curr. Med. Chem. 14 (2007) 1829e1852.
19] A.M. Gasco, A. Di Stilo, G. Sorba, A. Gasco, R. Ferioli, G. Folco, M. Civellic,
P. Carusoc, Eur. J. Med. Chem. 28 (1993) 433e438.
[20] S.J. Lewis, M.D. Travis, M.P. Hashmi-Hill, K. Sandock, T.P. Robertson, J.N. Bates,
2
009CB033), the National Key Tech Project for Major Creation of
[
New drugs (NO. 2009ZX09501-003) and the supports from the
School of Medicine, Zhejiang University for carrying out vaso-
relaxant effect assays.
Vasc. Pharmacol. 45 (2006) 383e394.
[
[
21] D.J. Triggle, D.A. Langs, R.A. Janis, Med. Res. Rev. 9 (1989) 123e180.
22] J.G. Hardman, L.E. Limbird, P.B. Molinoff, R.W. Ruddon, A.G. Gilman (Eds.), The
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