Design and stereoselective synthesis of novel isosteviol-fused pyrazolines and pyrazoles as potential anticancer agents

Article abstract of DOI:10.1016/j.ejmech.2013.04.044

Two series of novel isosteviol-fused pyrazoline and pyrazole derivatives were facilely synthesized via intramolecular 1,3-dipolar cycloaddition and condensation reaction, respectively. All compounds were characterized by NMR, IR and HRMS spectra. The stereochemistry of compounds 9b, 10, 11a and 11v were further confirmed by X-ray crystallographic analysis. The antiproliferative activities of the structurally related pyrazoline and pyrazole derivatives were tested in vitro on four human malignant cell lines (SGC 7901, A549, Raji and HeLa): Our results revealed that isosteviol-fused pyrazole derivatives exhibited noteworthy cytotoxic activities. Among them, 2,4-di-Cl-phenylpyrazole derivative 11t displayed better cytotoxities with IC50 values: 2.71, 3.18, 1.09 and 13.52 mM against SGC 7901, A549, Raji and HeLa, respectively, compared to cisplatin (IC₅₀ values: 7.56, 17.78, 17.32 and 14.31 μM, respectively).

Full text of DOI:10.1016/j.ejmech.2013.04.044

European Journal of Medicinal Chemistry 65 (2013) 70e82
Contents lists available at SciVerse ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Design and stereoselective synthesis of novel isosteviol-fused
pyrazolines and pyrazoles as potential anticancer agents
,
,
Song-Lin Zhu a, Ya Wu a b, Cong-Jun Liu c, Chang-Yong Wei c, Jing-Chao Tao c
,
*
,
Hong-Min Liu a
*
a New Drug Research & Development Center, School of Pharmaceutical Sciences, Zhengzhou University, No. 100, KeXue Avenue, Zhengzhou,
Henan 450001, China
b School of Pharmacy, Henan University of Traditional Chinese Medicine, Zhengzhou, Henan 450046, China
c College of Chemistry and Molecular Engineering, New Drug Research & Development Center, Zhengzhou University, 75 Daxue Road, Zhengzhou,
Henan 450052, China
a r t i c l e i n f o
a b s t r a c t
Article history:
Two series of novel isosteviol-fused pyrazoline and pyrazole derivatives were facilely synthesized via
intramolecular 1,3-dipolar cycloaddition and condensation reaction, respectively. All compounds were
characterized by NMR, IR and HRMS spectra. The stereochemistry of compounds 9b, 10, 11a and 11v were
further confirmed by X-ray crystallographic analysis. The antiproliferative activities of the structurally
related pyrazoline and pyrazole derivatives were tested in vitro on four human malignant cell lines (SGC
7901, A549, Raji and HeLa): Our results revealed that isosteviol-fused pyrazole derivatives exhibited
noteworthy cytotoxic activities. Among them, 2,4-di-Cl-phenylpyrazole derivative 11t displayed better
Received 25 March 2013
Received in revised form
19 April 2013
Accepted 20 April 2013
Available online 2 May 2013
Keywords:
Isosteviol
Stereoselective synthesis
Pyrazoline
cytotoxities with IC50 values: 2.71, 3.18, 1.09 and 13.52
respectively, compared to cisplatin (IC50 values: 7.56, 17.78, 17.32 and 14.31
m
M against SGC 7901, A549, Raji and HeLa,
M, respectively).
m
Ó 2013 Elsevier Masson SAS. All rights reserved.
Pyrazole
Cytotoxicity
1. Introduction
glucocorticoid agonist [11], antihypertension [12], antitumor [13],
antiproliferation [14] and other biologic activities [15e20]. Espe-
cially, Wu and co-workers reported that isosteviol derivatives with
exo-methylene cyclopentanone fragment had favorable cytotox-
icity [21,22]. Meanwhile, the 15- and/or 16-functionalized isosteviol
derivatives, obtained by means of group-conversion or structural
modification in our laboratory, had good cytotoxic activities
[23,24], which prompted us to further investigate new isosteviol
derivatives fused with heterocycles to develop novel stronger
anticancer agents for therapeutic use.
It is well known that a number of multicyclic compounds con-
taining heterocycle fragments exhibit a wide variety of biological
activities [25e31]. Pyrazoline and pyrazole are important structural
fragments of many bioactive compounds [32e34]. Kim and co-
workers revealed recently that compound A (Fig. 1) showed po-
Cancer is a leading cause of death worldwide, accounting for 8.7
million deaths (around 14% of all deaths) in 2012. Although many
chemotherapeutic agents, such as cisplatin, 5-fluorouracil and
taxol, have been developed to treat different kinds of cancer
effectively, some side effects could happen simultaneously. There-
fore, it is important and urgent to develop novel compounds as
anticancer agents with higher bioactivities and lower side effects
[1,2]. Functionalization of natural products with known pharma-
cophore moieties [3e5] and synthesis of their hybrid compounds
[6,7] are the most widely used approaches for obtaining novel
therapeutic agents in medicinal chemistry.
Isosteviol (ent-16-ketobeyeran-19-oic acid 1) is a tetracyclic
diterpenoid with a beyerane skeleton, obtained by acid hydrolysis
of stevioside [8,9]. In recent years, isosteviol derivatives have
attracted scientific attention because of their remarkably broad
spectrum of biological activities including anti-inflammatory [10],
tential ALK5 inhibitory activity as transforming growth factor-
b
type 1 receptor kinase inhibitors [35]. Synchronously, Janos wolf-
ling et al. reported that the cytotoxicity could be greatly improved
when p-methoxyphenylpyrazoline fragment was introduced to the
ring-D of steroid (Fig. 1, Compound B) [36]. In addition, the
carbothioamide-substituted pyrazole isosteviol derivative (Fig. 1,
Compound C), obtained in our laboratory, had good cytotoxic
* Corresponding authors. Tel./fax: þ86 371 67781739.
E-mail address: liuhm@zzu.edu.cn (H.-M. Liu).
0223-5234/$ e see front matter Ó 2013 Elsevier Masson SAS. All rights reserved.
http://dx.doi.org/10.1016/j.ejmech.2013.04.044

S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
71
OCH3
N
N
S
N
N
N
CN
N
OCH3
N
N
H
COOEt
HO
N
C
A
B
Fig. 1. Chemical structures of compounds AeC.
activities (IC50 values: 9.65, 17.73, 6.51 and 13.91
m
M against SGC
via intramolecular sequences [37]. Thus, the phenylhydrazones of
compound 5 as olefinic azomethine imine precursors were ex-
pected to undergo Lewis acid mediated 1,3-dipolar cycloadditions.
In this regard, a series of novel compounds containing a pyrazoline
ring fused with isosteviol structure were stereoselectively synthe-
sized via 1,3-dipolar cycloaddition (Scheme 1). Aldehyde 5 was
therefore initially reacted with phenylhydrazines 6ae6ad to
furnish the corresponding phenylhydrazones 7ae7ad. The ob-
tained compounds were readily underwent intramolecular cyclo-
addition in the presence of a catalytic amount of BF3$OEt2 to afford
pyrazoline derivatives 9ae9ad in 38%e81% yields. Compounds 7je
7l containing an electron-donating group on the aromatic ring
readily underwent heterocycle formation within 20 min to give 9je
9l in excellent yields. In contrast, the halogen-substituted phenyl-
hydrazone 7be7i and those containing an electron-withdrawing
group on the aromatic moiety (7ne7p) proved to be more stable,
and a longer reaction time was needed at the same temperature for
appreciable conversion.
7901, A549, Raji and HeLa cell lines, respectively). Based on these
facts and in continuation of our previous work, a series of novel
compounds containing pyrazole and pyrazoline ring fused with
isosteviol structure were designed and synthesized for the purpose
of new antitumor agent discovery.
2. Results and discussion
2.1. Chemistry
Isosteviol derivative 5 was synthesized as shown in Scheme 1
according to the procedure previously described [23]. Treatment
of isosteviol 1 with CH3CH2Br and KOH in DMSO afforded the cor-
responding ethyl ester of isosteviol 2 in 96% yield. Compound 3 was
stereoselectively synthesized via one pot Tollens’ reaction in 90%
yield. Treatment of compound 3 with 4-methylphenylsulfonyl
chloride in pyridine furnished compound 4 (75%), which was
further converted to the ring opening product 5 in 96% yield via
Grob fragmentation of compound 4 in the presence of NaOH in
CH3CN.
Compounds 9ae9ad were characterized by IR, 1H NMR, I3C NMR
and HRMS, respectively. The presence of signals at dH 4.24, 3.51,
2.97 and dC 173.0 in the NMR of compound 9b confirmed the for-
mation of pyrazoline ring. In addition, the relativity signals in the
The presence of the formyl group and the vinyl moiety in
compound 5 makes the molecule suitable for condensation with
hydrazine compounds and subsequent intramolecular 1,3-dipolar
cycloaddition to give fused heteroatom-containing frameworks
NOESY spectrum indicated the
a orientation of the protons at C-15
on D-ring of isosteviol. The stereostructure of 9b was further
confirmed by X-ray crystallographic analysis (Fig. 2).
Since pyrazole usually behaves as bioactive subunit in various
natural products, we intended to introduce the pyrazole fragment
to the skeleton of isosteviol in order to construct a novel family of
bioactive molecules for the discovery of antitumor agent. Mean-
while, the different effects of pyrazole and pyrazoline fragment in
the isosteviol molecules on antitumor activities could also be
investigated. Therefore, a series of pyrazole derivatives 11ae11ad
were designed and synthesized via the following procedure
(Scheme 2).
OH
O
ii
OH
OH
O
i
C
D
iii
OTs
A
1
B
COOEt
COOEt
COOEt
COOH
4
2
3
R
NHNH HCl
·
2
N
N
CHO
R
H
iv
6a-6ad
v
COOEt
COOEt
7a-7ad
5
_
R
BF
3H
R
N
N
N
+
vi
vi
N
COOEt
9a-9ad
COOEt
8a-8ad
9a: R = H ( 76%)
9k: R = m-CH3 ( 73%)
9l: R = p-CH3 ( 81%)
9u: R = 2,5-Cl2 ( 48%)
9v: R = 2,6-Cl2 ( 40%)
9b: R = o-F ( 58%)
9c: R = m-F ( 51%)
9d: R = p-F ( 54%)
9e: R = o-Cl ( 37%)
9f: R = m-Cl ( 76%)
9g: R = p-Cl ( 50%)
9h: R = m-Br ( 55%)
9i: R = p-Br ( 64%)
9j: R = o-CH3 ( 76%)
9m: R = p-OCH3 ( 51%) 9w: R = 3,4-Cl2 ( 58%)
9n :R = m-NO2 ( 38%)
9o: R = p-NO2 ( 49%)
9p: R = m-CF3 ( 48%)
9q: R = 2,4-F2 ( 65%)
9r: R = 2,5-F2 ( 42%)
9s: R = 2,3-Cl2( 41%)
9t: R = 2,4-Cl2 ( 43%)
9x: R = 3,5-Cl2 ( 59%)
9y: R = 2,3-Me2 ( 79%)
9z: R = 2,4-Me2( 75%)
9aa: R = 2,5-Me2 ( 62%)
9ab: R = 2,6-Me2( 41%)
9ac: R = 3,4-Me2 ( 73%)
9ad: R = 3,5-Me2 ( 46%)
Scheme 1. Reagents and conditions: (i) EtBr, DMSO, KOH, rt, 3 h, 96%; (ii) HCHO,
C2H5ONa, C2H5OH, 60 ꢀC, 3 h, 90%; (iii) TsCl, Pyr, rt, 18 h, 75%; (iv) NaOH, CH3CN, rt, 3 h,
96%; (v) 6ae6ad, sodium acetate, C2H5OH, reflux; (vi) toluene, BF3$OEt2, reflux, (38%e
81%).
Fig. 2. X-ray structure of compound 9b.

72
S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
R
NHNH HCl
·
2
O
OH
OH
R
N
N
i
6a-6ad
ii
OH
COOEt
11a-11ad
COOEt
COOEt
3
10
11a: R = H ( 76%)
11k:R = o-CH3 ( 56%)
11l: R = m-CH3 ( 87%)
11m:R = p-CH3 ( 89%)
11n: R = o-NO2 ( 61%)
11o :R = m-NO2 ( 89%)
11p: R = p-NO2 ( 57%)
11q: R = 2,4-F2( 87%)
11r: R = 2,5-F2 ( 88%)
11s: R = 2,3-Cl2 ( 92%)
11t: R = 2,4-Cl2 ( 90%)
11u: R = 2,5-Cl2 ( 76%)
11v: R = 2,6-Cl2 ( 56%)
11w: R = 3,4-Cl2 ( 75%)
11x: R = 3,5-Cl2( 60%)
11y: R = 2,3-Me2( 56%)
11z: R = 2,4-Me2 ( 63%)
11aa: R = 2,5-Me2 ( 79%)
11ab: R = 2,6-Me2 ( 69%)
11ac: R = 3,4-Me2 ( 59%)
11ad: R = 3,5-Me2 ( 81%)
11b: R = o-F ( 87%)
11c: R = m-F ( 93%)
11d: R = p-F ( 91%)
11e: R = o-Cl ( 87%)
11f: R = m-Cl ( 76%)
11g: R = p-Cl ( 50%)
11h: R = o-Br ( 55%)
11i: R = m-Br ( 64%)
11j: R = p-Br ( 76%)
Scheme 2. Reagents and conditions: (i)TCC, CH2Cl2, rt, 1.5 h, 62%; (ii) 6aead, isopropanol, reflux, (50%e91%).
Treatment of compound 3 with TCC in CH2Cl2 afforded the
The cell growth inhibitory potencies of the investigated com-
pounds, expressed as IC50 values (Table 1 and Table 2), demon-
strated that several of the derivatives exerted pronounced
antiproliferative effects on the four cell lines, which were similar to
or higher than those of cisplatin. Specifically, the pyrazole de-
rivatives 11ae11ad displayed much higher cytotoxicities than those
of the pyrazoline derivatives 9ae9ad, indicating that pyrazole
heterocyclic fragment may play an important role in their cytotoxic
activities.
As shown in Table 1, most of the pyrazoline derivatives 9ae9ad
showed better inhibitory activities against Raji cell than those in
SGC 7901, A549 and Hela cell lines. Interestingly, several of pyrazole
derivatives showed better inhibitory activities against Raji cell than
those of cisplatin. Meanwhile, compounds containing an electron-
donating group on the aromatic ring were more potent against
SGC 7901 cell than the halogen-substituted compounds (9l, 9m and
9ye9ac vs 9be9k and 9ne9x). Among the mono-substituted pyr-
azoline derivatives (9be9p), compound 9l with p-CH3 substitution
exhibited better cytotoxic activities with IC50 value of 29.39, 13.67,
corresponding product 10 in 62% yield. The structure of product 10
was confirmed unambiguously by the disappearance of the signals
at dH 3.98, 3.63 and 3.56 in the 1H NMR spectra, and the appearance
of signals at dH 12.09 and 7.15 in the 1H NMR and dC 215.1, 156.1,
121.3 in the 13C NMR spectra. The stereostructure of 10 was
confirmed by X-ray crystallographic analysis (Fig. 3), indicating that
the 15-formyl group profitably exists as its enol form.
Condensation of 10 with phenylhydrazines 6ae6ad in iso-
propanol provided the pyrazole derivatives 11ae11ad in 50%e91%
yields. The structures of compounds 11ae11ad were characterized
by IR, 1H NMR, I3C NMR and HRMS, respectively. In the 1H and 13
C
NMR spectra of 11v, additional resonances were observed at dH 7.48,
7.43, 7.34 and dC 153.3, 136.0, 135.8, 135.6, 135.2, 130.6, 129.6, 128.4,
128.3, suggesting the introduction of 2,6-di-Cl-phenylpyrazole
fragment. In addition, the stereostructure and regiochemistry of
11v and 11a were further confirmed by X-ray crystallographic
analysis (Fig. 4 and Fig. 5).
3.91 and 29.14
respectively.
mM against SGC 7901, A549, Raji and Hela cell lines,
2.2. Evaluation of cytotoxic activity
As shown in Table 2, all of the pyrazole derivatives 11ae11ad
had better cytotoxic activities than their precursor isosteviol. In
addition, most of pyrazole derivatives 11ae11ad, especially, meta-
substituted compounds, showed much higher inhibitory activities
against Raji cell than those in SGC 7901, A549 and Hela cell lines.
Meanwhile, among the mono-substituted pyrazole derivatives
(11be11p), NO2-substituted compounds (11ne11p) displayed
weaker cytotoxic activity than the other mono-substituted de-
rivatives whereas m-CH3-substituted pyrazole 11l displayed the
best cytotoxic activity. Among the di-substituted pyrazole de-
rivatives (11qe11ad), compounds 11ye11ad with two methyl
The in vitro cytotoxic activities of these compounds were then
evaluated against four cancer cell lines, including gastric cancer
(SGC 7901), lung cancer (A549), lympha cancer (Raji) and cervical
cancer (Hela) in comparison with cisplatin as the positive control.
Fig. 3. X-ray structure of compound 10.
Fig. 4. X-ray structure of compound 11v.

S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
73
Table 2
Cytotoxic activities of isosteviol-fused pyrazole derivatives in vitro.
Compound
R
Cytotoxic activities (IC50
,
mM)
SGC 7901
A549
Raji
Hela
1
>50
>50
>50
>50
11a
11b
11c
11d
11e
11f
11g
11h
11i
11j
11k
11l
11m
11n
11o
11p
11q
11r
H
o-F
m-F
p-F
o-Cl
m-Cl
p-Cl
o-Br
17.60
35.64
35.81
25.57
25.42
26.32
8.44
30.32
34.92
29.64
35.34
13.55
34.91
15.63
>50
>50
28.40
>50
>50
2.71
>50
27.11
>50
25.38
26.99
19.17
22.34
21.50
28.24
35.30
17.42
26.52
23.14
19.01
12.36
18.80
42.33
>50
>50
20.11
>50
>50
3.18
>50
33.16
>50
11.43
4.15
3.40
7.01
10.11
9.54
13.82
11.35
3.27
4.78
7.71
4.84
8.61
14.35
7.40
10.15
9.96
7.11
5.13
1.09
11.72
5.70
17.13
15.85
3.26
6.55
7.94
2.09
6.50
12.49
17.32
42.45
>50
>50
>50
34.65
>50
>50
41.10
>50
>50
15.15
>50
>50
>50
>50
>50
>50
>50
>50
13.52
>50
34.93
>50
>50
37.45
43.29
34.33
20.34
38.17
>50
m-Br
p-Br
o-CH3
m-CH3
p-CH3
o-NO2
m-NO2
p-NO2
2,4-F2
2,5-F2
2,3-Cl2
2,4-Cl2
2,5-Cl2
2,6-Cl2
3,4-Cl2
3,5-Cl2
2,3-Me2
2,4-Me2
2,5-Me2
2,6-Me2
3,4-Me2
3,5-Me2
Fig. 5. X-ray structure of compound 11a.
substituents showed much higher cytotoxic activities than the
other di-substituted pyrazole derivatives. Especially, compound
11ab, with 2,6-dimethyl phenyl as the aromatic substituent, was
proved to be an outstandingly potent cytotoxic compound on the
four cell lines. More importantly, all of pyrazole derivatives (11ae
11ad) showed superior cytotoxic activities to Cisplatin resistant Raji
cell. Specifically, pyrazole derivative 11t exhibited noteworthy
11s
11t
11u
11v
11w
11x
11y
11z
11aa
11ab
11ac
11ad
Cisplatin
>50
>50
19.33
18.70
32.96
9.17
38.06
>50
18.17
16.52
17.06
11.81
15.76
>50
cytotoxic activities with IC50 value: 2.71, 3.18, 1.09 and 13.52 mM
against SGC 7901, A549, Raji and Hela cell lines, respectively, and all
superior than that of Cisplatin. The currently known structuree
activity relationships offer a promising possibility for further
rational design of more derivatives.
7.56
17.78
14.31
3. Conclusion
Table 1
Cytotoxic activities of isosteviol-fused pyrazoline derivatives in vitro.
In summary, two series of novel isosteviol derivatives containing
pyrazoline and pyrazole heterocyclic fragments were stereo-
selectively synthesized in highyields by functional interconversions
in ring D of the tetracyclic diterpene isosteviol. The in vitro cytotoxic
activities against four human tumor cell lines were evaluated. The
results revealed that introduction of pyrazoline and pyrazole het-
erocyclic fragments to isosteviol were beneficial to the cytotoxic
activities. Furthermore, pyrazole derivatives 11ae11ad showed su-
perior cytotoxic activities to cisplatin against Raji cell. Specifically,
Compound
R
Cytotoxic activities (IC50
,
m
M)
Raji
SGC 7901
A549
Hela
1
9a
9b
9c
9d
9e
9f
9g
9h
9i
9j
>50
>50
34.67
>50
>50
>50
>50
>50
>50
>50
>50
>50
29.39
34.02
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
31.85
28.49
22.54
16.61
32.49
>50
7.56
>50
28.33
32.62
>50
>50
>50
>50
>50
>50
>50
>50
>50
13.67
17.84
>50
>50
>50
>50
20.21
>50
>50
>50
11.57
>50
>50
>50
>50
>50
17.55
>50
>50
17.78
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
29.14
31.21
>50
>50
>50
>50
>50
>50
>50
>50
>50
>50
24.55
>50
>50
30.32
28.77
>50
>50
14.31
H
o-F
m-F
p-F
o-Cl
m-Cl
p-Cl
m-Br
13.95
17.23
29.10
4.51
>50
4.57
>50
25.05
>50
19.89
>50
3.91
compound 11t (IC50 values: 2.71, 3.18,1.09 and 13.52 mM against SGC
7901, A549, Raji and HeLa, respectively) had the most potent cyto-
toxicity, which may be exploitable as a lead compound for the
development of potent antitumor agents. Further efforts aiming at
developing potent anticancer agents based on appropriately
modified D-ring fused heterocyclic analogs are continuing in our
laboratory, which will be reported in due course.
p-Br
o-CH3
m-CH3
p-CH3
p-OCH3
m-NO2
p-NO2
m-CF3
2,4-F2
2,5-F2
2,3-Cl2
2,4-Cl2
2,5-Cl2
2,6-Cl2
3,4-Cl2
3,5-Cl2
2,3-Me2
2,4-Me2
2,5-Me2
2,6-Me2
3,4-Me2
3,5-Me2
9k
9l
9m
9n
9o
9p
9q
9r
9s
9t
9u
9v
9w
9x
12.25
>50
>50
7.21
4. Experimental
13.53
27.36
22.14
22.87
25.63
>50
>50
>50
8.82
11.18
12.63
5.63
8.36
>50
4.1. General methods
All reagents and solvents were obtained from commercial sup-
pliers. All the reactions were monitored by TLC. Melting points
were determined on a Beijing Keyi XT5 apparatus and the tem-
perature was not corrected. IR spectra were recorded as KBr pellets
on a Thermo Nicolet (IR200) spectrometer. 1H and 13C NMR spectra
were recorded on a Bruker DPX-400 spectrometer at 400 and
100 MHz, respectively, with TMS as internal standard. Mass spectra
were taken by Waters Q-Tof micro mass spectrometer. X-ray anal-
ysis was taken on a Rigaku RAXIS-IV.
9y
9z
9aa
9ab
9ac
9ad
Cisplatin
17.32

74
S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
4.2. General procedure for cytotoxicity assay
(ESI, m/z) calcd for C23H38O4Na [M þ Na]þ 401.2668. Found:
401.2664.
In vitro cytotoxicity study: Cell lines of human gastric cancer
SGC-7901, human lung adenocarcinoma A549 and human cervical
carcinoma Hela were purchased from the Type Culture Collection of
the Chinese Academy of Sciences, Shanghai, China. Human B lym-
phoma Raji ecll line was obtained from China Center for Type
Culture Collection, Wuhan, China. Cells were cultured in RPMI-1640
4.3.4. Ethyl ent-15
beyeran-19-oate (4)
a-(4-toluenesulfonyl)-oxymethyl-16b-hydroxy
A mixture of compound 3(0.378 g, 1 mmol) and 4-methyl
phenylsulfonyl chloride (0.238 g, 1.1 mmol) in pyridine (5 mL)
was stirred at room temperature for 18 h. Then the reaction mixture
was filtered, and the filtrate was extracted with CH2Cl2 and HCl
aqueous solution (5 M). At last, the organic layer was washed with
saturated NaCl aqueous solution, dried with MgSO4 and the filtrate
was concentrated. The residue was purified by column chroma-
tography on silica (petroleum ether/ethyl acetate ¼ 4:1, v/v) to give
product 4 (0.399 g, 75%). IR (KBr): 3541, 2950, 2928, 2851, 1718,
1598, 1458, 1361, 1177, 1151, 1097, 1020, 948, 924, 816, 779, 665,
medium with 10% FBS, 100 U/mL of penicillin and 100 mg/mL of
streptomycin in a 5% CO2 humidified atmosphere at 37 ꢀC. Cell
cytotoxicity was assayed by MTT method. Briefly, cells were seeded
in 96-well tissue culture plates and incubated with compounds (0e
50 m
mol/L) for 48 h at 37 ꢀC, 5% CO2. Following treatment, MTT
(0.5 mg/mL) was added. After an additional 4 h of incubation, the
reaction was terminated by removal of the supernatant. Then
200
m
l of DMSO was added to each well to dissolve the formazan
555 cmꢁ1
;
1H NMR (400 MHz, CDCl3, ppm):
d
7.82 (d, J ¼ 8.2 Hz,
product. Optical density (OD) of each well was measured at 570 nm
with PowerWaveX Microplate Scanning Spectrophotometer (Bio-
tek Instruments, Inc). Then the inhibitory percentage of each
compound at various concentrations to the cell proliferation was
calculated.
2H), 7.37 (t, J ¼ 8.2 Hz, 1H), 4.32 (dd, J ¼ 9.8, 3.6 Hz, 1H), 4.07 (q,
J ¼ 7.1 Hz, 2H), 3.96 (t, J ¼ 9.8 Hz, 1H), 3.47 (d, J ¼ 7.5 Hz,1H), 3.46 (s,
3H), 2.33 (m, 1H), 2.18 (m, 2H), 1.80e1.28 (m, 8H), 1.23 (t, J ¼ 7.1 Hz,
3H), 1.19e1.11 (m, 1H), 1.16 (s, 3H), 1.10e0.93 (m, 5H), 0.88 (s, 3H),
0.86e0.79 (m, 3H), 0.67 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d
177.2, 144.8, 132.9, 129.8, 129.8, 127.7, 127.7, 84.9, 72.8, 59.9, 57.4,
4.3. The preparation of isosteviol derivatives
56.8, 53.8, 47.6, 43.5, 42.8, 40.9, 39.5, 38.0, 37.8, 34.6, 33.0, 28.8,
24.8, 21.9, 21.6, 19.3, 18.7, 14.0, 12.9; HRMS (ESI, m/z) calcd for
4.3.1. Ent-16-oxobeyeran-19-oic acid (1) [9]
C
30H44O6SNa [M þ Na]þ 555.2757. Found: 555.2742.
Isosteviol 1 was synthesized by hydrolysis of stevioside with
dilute sulfuric acid. Mp 228e230 ꢀC; IR (KBr): 3455, 2954, 2927,
2852, 2679, 1738, 1697, 1474, 1453, 1406, 1372, 1320, 1271, 1238,
4.3.5. Ring opening product (5)
A mixture of compound 4 (0.532 g, 1 mmol) and NaOH (0.048 g,
1.1 mmol) in dry CH3CN (5 mL) was stirred at room temperature for
3 h. Then the reaction mixture was filtered, the filtrate was
concentrated, and the residue was extracted with CH2Cl2 and H2O.
At last, the organic layer was washed with saturated NaCl aqueous
solution, dried with MgSO4 and the filtrate was concentrated. The
residue was crystallized from CHCl3 to give product 5 (0.345 g, 96%).
Mp 116.5e117.8 ꢀC; IR (KBr): 3072, 2937, 2796, 2704, 1716, 1458,
1179, 950, 797 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
; d 2.64 (dd,
J ¼ 18.6, 3.8 Hz, 1H), 2.17 (d, J ¼ 13.4 Hz, 1H), 1.90e1.82 (m, 2H), 1.81
(d, J ¼ 18.6 Hz 1H), 1.77e1.38 (m, 10H), 1.26 (s, 3H), 1.22e1.14 (m,
3H), 1.07e0.99 (m, 1H), 0.98 (s, 3H), 0.95e0.88 (m, 1H), 0.79 (s, 3H);
HRMS (ESI, m/z) calcd for C20H30O3Na [M þ Na]þ 341.2093. Found:
341.2085.
4.3.2. Ethyl ent-16-oxobeyeran-19-oate (2)
1384, 1238, 1183, 1029, 912, 704 cmꢁ1
ppm):
;
1H NMR (400 MHz, CDCl3,
The isosteviol ethyl ester 2 was obtained by treating isosteviol
with CH3CH2Br and KOH in DMSO at room temperature in 92% yield
according to the literature method [38]. Mp 125e127 ꢀC; IR (KBr):
2957, 2926, 2847, 1726, 1451, 1377, 1227, 1146, 1096, 1024 cmꢁ1; 1H
d
9.27 (d, J ¼ 1.6 Hz, 1H), 5.95 (dd, J ¼ 17.6, 10.8 Hz, 1H), 5.12
(d, J ¼ 10.0 Hz, 1H), 5.08 (d, J ¼ 3.2 Hz, 1H), 4.01 (m, 2H), 2.28 (m,
1H), 2.13 (m, 2H), 1.88e1.39 (m, 9H), 1.26 (d, J ¼ 13.2 Hz, 1H), 1.20 (t,
J ¼ 7.2 Hz, 3H), 1.14 (s, 3H), 1.10e0.89 (m, 5H), 0.88 (s, 3H), 0.58 (s,
NMR (400 MHz, CDCl3, ppm):
d
4.10 (q, J ¼ 7.2 Hz, 2H), 2.96 (dd,
3H); 13C NMR (100 MHz, CDCl3, ppm):
d 205.2, 177.3, 142.5, 113.6,
J ¼ 18.4, 2.8 Hz, 1H), 2.17 (d, J ¼ 11.2 Hz, 1H), 1.99 (d, J ¼ 18.8 Hz, 1H),
1.88e1.58 (m, 8H), 1.47e1.28 (m, 6H), 1.26 (t, J ¼ 7.2 Hz, 3H), 1.24e
1.19 (m, 2H), 1.18 (s, 3H), 1.07 (s, 3H), 1.05e0.84 (m, 1H), 0.77 (s, 3H);
HRMS (ESI, m/z) calcd for C22H34O3Na [M þ Na]þ 369.2406. Found:
369.2400.
59.9, 57.7, 57.5, 55.2, 46.1, 43.7, 40.8, 40.1, 39.6, 38.1, 38.0, 32.3, 28.7,
24.6, 20.0, 19.1, 17.4, 14.0, 13.2; HRMS (ESI, m/z) calcd for
C
23H36O3Na [M þ Na]þ 383.2562. Found: 383.2560.
4.3.6. General procedure for synthesis of compounds 9ae9ad
A mixture of compound 5 (0.360 g, 1 mmol) and various phe-
nylhydrazines 6ae6ad (0.145e0.225 g,1 mmol) in dry EtOH (10 mL)
were stirred at 78 ꢀC in the presence of CH3COONa (0.099 g,
1.2 mmol). After completion of the reaction monitored by TLC, the
solution was then poured into water (10 mL), the white precipitate
was filtered and washed with water and dried. The crude product
was recrystallized from ethyl acetate/light petroleum ether to give
compounds 7ae7ad. Then to a solution of compounds 7ae7ad
(0.450e0.529 g, 1 mmol) in toluene (5 mL), 48% BF3$OEt2 (48%
solution in Et2O, 0.05 mL, 0.17 mmol) was added dropwise and the
mixture was heated under a nitrogen atmosphere at 118 ꢀC. After
completion of the reaction monitored by TLC, water (10 mL) was
added to the mixture, which was next neutralized with NaHCO3,
and the organic phase was separated and dried over Na2SO4. After
evaporation in vacuo, the crude product was purified by column
chromatography to give product 9ae9ad.
4.3.3. Ethyl ent-15a-hydroxymethyl-16b-hydroxybeyeran-19-oate
(3) [38]
To a stirred solution of compound 2 (0.346 g, 1 mmol) and
C2H5ONa (0.136 g, 2 mmol) in ethanol (20 mL) was added 37%
formaldehyde aqueous solution (2 mL). After stirring for 3 h at
60 ꢀC, the mixture was concentrated under vacuum, and extrac-
ted with CHCl3 and H2O, at last the organic layer was washed with
saturated NaCl aqueous solution, dried with MgSO4 and concen-
trated under vacuum to give white powder 8 (0.34 g, 90%). Mp
181e182 ꢀC; IR (KBr): 3435, 2940, 2838, 1720, 1458, 1378, 1234,
1179, 1153, 1123 cmꢁ1; 1H NMR (400 MHz, CDCl3, ppm):
d 4.09 (q,
J ¼ 7.2 Hz, 2H), 3.98 (dd, J ¼ 9.7, 5.0 Hz, 1H), 3.63 (d, J ¼ 4.7 Hz,
1H), 3.56 (t, J ¼ 10.2 Hz, 1H), 2.16 (d, J ¼ 13.0 Hz, 1H), 2.05 (m, 1H),
1.83e1.56 (m, 9H), 1.43e1.37 (m, 2H), 1.26 (t, J ¼ 7.2 Hz, 3H), 1.22e
1.19 (m, 1H), 1.16 (s, 3H), 1.08e0.95 (m, 4H), 0.94 (s, 3H), 0.88e
0.86 (m, 1H), 0.78 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d
177.4, 86.7, 64.9, 60.0, 57.5, 57.0, 54.2, 50.2, 43.6, 42.4, 40.8, 39.6,
4.3.6.1. Pyrazoline derivative 9a. Yield 76%; Mp 98.3e100.2 ꢀC; IR
38.1, 37.9, 34.8, 33.0, 28.9, 25.0, 22.1, 19.5, 18.8, 14.1, 13.2; HRMS
(KBr): 3054, 2926, 2849, 1720, 1599, 1497, 1454, 1378, 1323, 1292,

S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
75
1259, 1229, 1175, 1151, 1092, 1027, 991, 852, 751, 693, 517 cmꢁ1; 1H
NMR (400 MHz, CDCl3, ppm):
1H), 2.74 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.21e2.17 (m, 1H), 1.84e1.68 (m,
5H), 1.65e1.53 (m, 4H), 1.46e1.31 (m, 3H), 1.27 (t, J ¼ 7.2 Hz, 3H),
1.25 (s, 3H),1.21e1.20 (m,1H),1.17 (s, 3H),1.13e0.87 (m, 4H), 0.84 (s,
d
7.28 (d, J ¼ 8.2 Hz, 2H), 7.12 (d,
J ¼ 7.6 Hz, 2H), 6.86 (t, J ¼ 7.2 Hz, 1H), 4.12 (q, J ¼ 7.2 Hz, 2H), 3.98 (t,
J ¼ 8.6 Hz, 1H), 3.51e3.46 (m, 1H), 2.99 (dd, J ¼ 14.0, 8.6 Hz, 1H),
2.21e2.18 (m, 1H), 1.90e1.70 (m, 5H), 1.67e1.54 (m, 4H), 1.45e1.33
(m, 2H), 1.28 (t, J ¼ 7.2 Hz, 3H), 1.25e1.21 (m, 2H), 1.19 (s, 6H), 1.16e
3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2, 173.9, 147.7, 130.2,
127.5, 126.2, 124.5, 122.1, 60.5, 60.0, 58.7, 57.2, 56.8, 50.5, 43.6, 42.2,
39.9, 39.8, 38.2, 37.8, 35.2, 35.2, 29.0, 21.7, 21.6, 19.7, 18.8, 14.0, 13.3;
HRMS (ESI, m/z) calcd for C29H40ClN2O2 [M þ H]þ 483.2778. Found:
483.2773; [Mþ2 þ H]þ 485.2778. Found: 485.2771.
0.86 (m, 4H), 0.82 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2,
172.2, 149.1, 128.9, 128.9, 119.7, 114.3, 114.3, 60.0, 58.6, 57.2, 56.7,
55.2, 49.7, 43.6, 42.0, 39.7, 39.7, 38.2, 37.8, 35.4, 35.2, 29.0, 21.8, 21.3,
19.7, 18.8, 14.1, 13.1; HRMS (ESI, m/z) calcd for C29H41N2O2 [M þ H]þ
449.3168. Found: 449.3170.
4.3.6.6. Pyrazoline derivative 9f. Yield 76%; Mp 143.4e144.6 ꢀC; IR
(KBr): 3055, 2946, 2850, 1712, 1622, 1595, 1567, 1485, 1451, 1375,
1342, 1260, 1242, 1181, 1153, 1096, 1033, 992, 852, 827, 758,
4.3.6.2. Pyrazoline derivative 9b. Yield 58%; Mp 124.2e125.8 ꢀC; IR
(KBr): 3067, 2953, 2848, 1720, 1635, 1610, 1495, 1455, 1379, 1320,
1293, 1253, 1229, 1176, 1149, 1101, 1027, 976, 849, 818, 749,
676 cmꢁ1
;
1H NMR (400 MHz, CDCl3, ppm):
d
7.16 (t, J ¼ 8.0 Hz,
1H), 7.10 (t, J ¼ 2.0 Hz, 1H), 6.95e6.92 (m, 1H), 6.81e6.79 (m, 1H),
4.12 (q, J ¼ 7.2 Hz, 2H), 3.94 (t, J ¼ 8.6 Hz, 1H), 3.50e3.44 (m, 1H),
2.99 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.21e2.18 (m, 1H), 1.90e1.73 (m,
5H), 1.65e1.53 (m, 4H), 1.46e1.34 (m, 2H), 1.28 (t, J ¼ 7.2 Hz, 3H),
1.25e1.21 (m, 2H), 1.19 (s, 3H), 1.19 (s, 3H), 1.16e0.89 (m, 4H), 0.81
562 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
; d 7.58e7.53 (m, 1H),
7.07e7.01 (m, 2H), 6.92e6.87 (m, 1H), 4.26e4.20 (m, 1H), 4.12 (q,
J ¼ 7.2 Hz, 2H), 3.53e3.48 (m, 1H), 2.97 (dd, J ¼ 14.4, 8.6 Hz, 1H),
2.20e2.17 (m, 1H), 1.87e1.73 (m, 5H), 1.65e1.53 (m, 4H), 1.45e1.35
(m, 2H),1.28 (t, J ¼ 7.2 Hz, 3H),1.25e1.23 (m, 2H),1.22 (s, 3H),1.18 (s,
3H), 1.16e0.89 (m, 4H), 0.83 (s, 3H); 13C NMR (100 MHz, CDCl3,
(s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2, 172.4,
150.0, 134.6, 129.9, 119.2, 114.1, 112.2, 60.0, 58.5, 57.1, 56.7, 54.7,
49.7, 43.6, 42.1, 39.7, 39.7, 38.2, 37.8, 35.3, 35.2, 29.0, 21.8, 21.3,
19.7, 18.8, 14.2, 13.1; HRMS (ESI, m/z) calcd for C29H40ClN2O2
[M þ H]þ 483.2778. Found: 483.2777; [Mþ2 þ H]þ 485.2778.
Found: 485.2785.
ppm):
d
177.2, 173.0, 152.5 (d, J ¼ 242.3 Hz), 137.9 (d, J ¼ 8.8 Hz),
124.3 (d, J ¼ 3.4 Hz), 121.9 (d, J ¼ 7.4 Hz), 119.6 (d, J ¼ 3.2 Hz), 116.0
(d, J ¼ 20.5 Hz), 60.0, 58.9 (d, J ¼ 6.6 Hz), 57.1, 56.7, 53.4, 50.3, 43.6,
42.1, 39.8, 39.8, 38.2, 37.8, 35.3, 35.2, 29.0, 21.8, 21.4, 19.7, 18.8, 14.1,
13.2; HRMS (ESI, m/z) calcd for C29H40FN2O2 [M þ H]þ 467.3074.
Found: 467.3076.
4.3.6.7. Pyrazoline derivative 9g. Yield 50%; Mp 87.9e89.6 ꢀC; IR
(KBr): 3052, 2953, 2848, 1720, 1628, 1596, 1492, 1456, 1378, 1322,
1261, 1228, 1150, 1093, 1029, 976, 851, 821, 496 cmꢁ1 1H NMR
;
4.3.6.3. Pyrazoline derivative 9c. Yield 51%; Mp 80.8e82.5 ꢀC; IR
(KBr): 3077, 2953, 2849, 1720, 1611, 1581, 1492, 1454, 1378, 1344,
1322, 1296, 1267, 1229, 1180, 1149, 1094, 1029, 927, 852, 763, 681,
(400 MHz, CDCl3, ppm): d 7.22e7.19 (m, 2H), 7.03e7.00 (m, 2H), 4.12
(q, J ¼ 7.2 Hz, 2H), 3.94 (t, J ¼ 8.6 Hz, 1H), 3.50e3.44 (m, 1H), 2.95
(dd, J ¼ 14.4, 8.6 Hz,1H), 2.21e2.18 (m,1H),1.90e1.73 (m, 5H),1.70e
1.52 (m, 4H), 1.45e1.35 (m, 2H), 1.28 (t, J ¼ 7.2 Hz, 3H), 1.25e1.21 (m,
2H), 1.19 (s, 3H), 1.18 (s, 3H), 1.16e0.89 (m, 4H), 0.81 (s, 3H); 13C
522 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
; d 7.21e7.16 (m, 1H),
6.87e6.83 (m, 1H), 6.80e6.78 (m, 1H), 6.55e6.50 (m, 1H), 4.12 (q,
J ¼ 7.2 Hz, 2H), 3.94 (t, J ¼ 8.6 Hz, 1H), 3.50e3.44 (m, 1H), 3.00 (dd,
J ¼ 14.4, 8.6 Hz,1H), 2.21e2.18 (m, 1H), 1.90e1.73 (m, 5H),1.66e1.52
(m, 4H),1.45e1.34 (m, 2H),1.28 (t, J ¼ 7.2 Hz, 3H),1.25e1.20 (m, 2H),
1.20 (s, 3H), 1.19 (s, 3H), 1.17e0.89 (m, 4H), 0.81 (s, 3H); 13C NMR
NMR (100 MHz, CDCl3, ppm): d 177.2,172.4,147.7,128.7,128.7,124.3,
115.3, 115.3, 60.0, 58.5, 57.1, 56.7, 55.0, 49.8, 43.6, 42.0, 39.7, 39.7,
38.2, 37.8, 35.4, 35.2, 29.0, 21.8, 21.3, 19.7, 18.8, 14.1, 13.1; HRMS (ESI,
m/z) calcd for C29H40ClN2O2 [M þ H]þ 483.2778. Found: 483.2775;
[Mþ2 þ H]þ 485.2778. Found: 485.2791.
(100 MHz, CDCl3, ppm):
d
177.2, 172.3, 163.6 (d, J ¼ 241.3 Hz), 150.6
(d, J ¼ 10.6 Hz), 130.0 (d, J ¼ 9.7 Hz), 109.4 (d, J ¼ 2.2 Hz), 105.9 (d,
J ¼ 21.6 Hz), 101.5 (d, J ¼ 28.8 Hz), 60.0, 58.5, 57.1, 56.7, 54.7, 49.7,
43.6, 42.0, 39.7, 39.7, 38.2, 37.8, 35.4, 35.2, 29.0, 21.8, 21.3, 19.7, 18.8,
14.2, 13.1; HRMS (ESI, m/z) calcd for C29H40FN2O2 [M þ H]þ
467.3074. Found: 467.3077.
4.3.6.8. Pyrazoline derivative 9h. Yield 55%; Mp 141.5e143.2 ꢀC; IR
(KBr): 3062, 2953, 2848, 1720, 1590, 1560, 1479, 1377, 1339, 1323,
1229, 1178, 1150, 1095, 1029, 985, 851, 766, 681 cmꢁ1 1H NMR
;
(400 MHz, CDCl3, ppm):
d
7.26e7.24 (m, 1H), 7.10 (t, J ¼ 8.0 Hz, 1H),
6.99e6.94 (m, 2H), 4.12 (q, J ¼ 7.2 Hz, 2H), 3.93 (t, J ¼ 8.6 Hz, 1H),
3.49e3.44 (m, 1H), 2.98 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.21e2.18 (m, 1H),
1.90e1.73 (m, 5H), 1.65e1.52 (m, 4H), 1.45e1.34 (m, 2H), 1.28 (t,
J ¼ 7.2 Hz, 3H), 1.25e1.22 (m, 2H), 1.19 (s, 3H), 1.18 (s, 3H), 1.16e0.86
4.3.6.4. Pyrazoline derivative 9d. Yield 54%; Mp 114.8e116.5 ꢀC; IR
(KBr): 3053, 2951, 2848, 1720, 1630, 1606, 1506, 1452, 1379, 1323,
1294, 1221, 1177, 1151, 1098, 1028, 976, 850, 825, 517 cmꢁ1; 1H NMR
(400 MHz, CDCl3, ppm):
d
7.07e7.03 (m, 2H), 6.98e6.94 (m, 2H),
(m, 4H), 0.81 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2,
4.12 (q, J ¼ 7.2 Hz, 2H), 3.93 (t, J ¼ 8.6 Hz, 1H), 3.50e3.44 (m, 1H),
2.92 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.21e2.18 (m, 1H), 1.90e1.70 (m, 5H),
1.67e1.52 (m, 4H), 1.45e1.34 (m, 2H), 1.28 (t, J ¼ 7.2 Hz, 3H), 1.25e
1.21 (m, 2H), 1.19 (s, 3H), 1.18 (s, 3H), 1.161e0.89 (m, 4H), 0.81 (s,
172.5, 150.1, 130.2, 122.9, 122.1, 116.9, 112.7, 60.0, 58.5, 57.1, 56.7,
54.6, 49.7, 43.6, 42.1, 39.7, 39.7, 38.2, 37.8, 35.3, 35.2, 29.0, 21.8, 21.3,
19.7, 18.8, 14.2, 13.1; HRMS (ESI, m/z) calcd for C29H40BrN2O2
[M þ H]þ 527.2273. Found: 527.2277; [Mþ2 þ H]þ 529.2273.
Found: 529.2250.
3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2, 172.3, 157.2 (d,
J ¼ 236.2 Hz), 145.8 (d, J ¼ 2.0 Hz), 115.4 (d, J ¼ 7.7 Hz), 115.4 (d,
J ¼ 7.7 Hz), 115.3 (d, J ¼ 22.4 Hz), 115.3 (d, J ¼ 22.4 Hz), 60.0, 58.6,
57.1, 56.7, 55.9, 50.0, 43.6, 42.0, 39.7, 39.7, 38.2, 37.8, 35.4, 35.2, 29.0,
21.8, 21.3, 19.7, 18.8, 14.1, 13.1; HRMS (ESI, m/z) calcd for
4.3.6.9. Pyrazoline derivative 9i. Yield 64%; Mp 90.5e92.2 ꢀC; IR
(KBr): 3059, 2952, 2848, 1720, 1628, 1590, 1489, 1454, 1378, 1322,
1293, 1261, 1229, 1176, 1151, 1096, 1029, 976, 852, 819, 511 cmꢁ1; 1H
C
29H40FN2O2 [M þ H]þ 467.3074. Found: 467.3078.
NMR (400 MHz, CDCl3, ppm): d 7.36e7.32 (m, 2H), 6.98e6.95 (m,
2H), 4.11 (q, J ¼ 7.2 Hz, 2H), 3.93 (t, J ¼ 8.6 Hz, 1H), 3.50e3.44 (m,
1H), 2.95 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.21e2.17 (m, 1H), 1.90e1.72 (m,
5H), 1.65e1.52 (m, 4H), 1.46e1.35 (m, 2H), 1.28 (t, J ¼ 7.2 Hz, 3H),
1.25e1.21 (m, 2H),1.19 (s, 3H),1.18 (s, 3H),1.15e0.89 (m, 4H), 0.81 (s,
4.3.6.5. Pyrazoline derivative 9e. Yield 37%; Mp 190.5e192.3 ꢀC; IR
(KBr): 3057, 2956, 2842, 1717, 1637, 1588, 1514, 1469, 1378, 1319,
1282, 1249, 1227, 1148, 1092, 1039, 1016, 978, 848, 752, 671 cmꢁ1
;
1H NMR (400 MHz, CDCl3, ppm):
d
7.52 (dd, J ¼ 8.0, 1.4 Hz, 1H), 7.35
3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2, 172.4, 148.1, 131.6,
(dd, J ¼ 8.0, 1.4 Hz, 1H), 7.22 (dt, J ¼ 7.2, 1.4 Hz, 1H), 7.01 (dt, J ¼ 7.2,
131.6, 115.8, 115.8, 111.5, 60.0, 58.5, 57.1, 56.7, 54.9, 49.8, 43.6, 42.0,
39.7, 39.7, 38.2, 37.8, 35.3, 35.2, 29.0, 21.8, 21.3, 19.7, 18.8, 14.2, 13.1;
1.4 Hz, 1H), 4.33 (t, J ¼ 8.6 Hz, 1H), 4.16e4.07 (m, 2H), 3.57e3.51 (m,

76
S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
HRMS (ESI, m/z) calcd for C29H40BrN2O2 [M þ H]þ 527.2273. Found:
527.2271; [Mþ2 þ H]þ 529.2273. Found: 529.2241.
2.22e2.18 (m, 1H), 1.92e1.73 (m, 5H), 1.67e1.53 (m, 4H), 1.46e1.37
(m, 2H), 1.29 (t, J ¼ 7.2 Hz, 3H), 1.27e1.22 (m, 2H), 1.20 (s, 6H), 1.16e
0.94 (m, 4H), 0.82 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2,
4.3.6.10. Pyrazoline derivative 9j. Yield 76%; Mp 131.1e131.8 ꢀC; IR
(KBr): 3062, 2952, 2848, 1720, 1630, 1598, 1491, 1454, 1378, 1321,
1254, 1231, 1178, 1151, 1095, 1027, 977, 850, 756, 718 cmꢁ1; 1H NMR
173.2, 149.4, 149.0, 129.5, 120.0, 113.7, 108.0, 60.1, 58.4, 57.1, 56.6,
54.4, 49.8, 43.6, 42.1, 39.7, 39.7, 38.2, 37.8, 35.3, 35.2, 29.0, 21.8, 21.3,
19.7, 18.8, 14.2, 13.1; HRMS (ESI, m/z) calcd for C29H40N3O4 [M þ H]þ
494.3019. Found: 494.3021.
(400 MHz, CDCl3, ppm):
d
7.21e7.13 (m, 3H), 6.98 (dt, J ¼ 14.4,
8.6 Hz,1H), 4.13e4.07 (m, 2H), 3.78 (t, J ¼ 8.6 Hz,1H), 3.48e3.43 (m,
1H), 2.83 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.41 (s, 3H), 2.20e2.17 (m, 1H),
1.85e1.73 (m, 5H), 1.67e1.55 (m, 4H), 1.45e1.29 (m, 3H), 1.26 (t,
J ¼ 7.2 Hz, 3H), 1.24e1.21 (m, 1H), 1.21 (s, 3H), 1.17 (s, 3H), 1.14e0.89
4.3.6.15. Pyrazoline derivative 9o. Yield 49%; Mp 186.8e188.7 ꢀC; IR
(KBr): 2953, 2927, 2851, 1720, 1595, 1502, 1456, 1383, 1317, 1179,
1109, 839 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
; d 8.14 (d,
(m, 4H), 0.84 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d
177.2,
J ¼ 9.2 Hz, 2H), 6.98 (d, J ¼ 9.2 Hz, 2H), 4.13 (m, 2H), 4.06 (m, 1H),
3.54 (t, J ¼ 17.2 Hz, 1H), 3.25 (dd, J ¼ 13.6, 9.2 Hz, 1H), 2.26 (m, 3H),
1.92e1.73 (m, 4H), 1.68e1.30 (m, 7H), 1.30 (t, J ¼ 7.2 Hz, 3H), 1.20 (s,
3H), 1.18 (s, 3H), 1.16e0.83 (m, 4H), 0.80 (s, 3H); 13C NMR (100 MHz,
172.8, 148.9, 131.0, 130.9, 126.4, 123.5, 118.8, 60.0, 59.7, 58.8, 57.2,
56.8, 50.2, 43.6, 42.0, 39.8, 39.8, 38.2, 37.8, 35.3, 35.3, 29.0, 21.8, 21.5,
19.8, 19.2, 18.8, 14.1, 13.3; HRMS (ESI, m/z) calcd for C30H43N2O2
[M þ H]þ 463.3325. Found: 463.3326.
CDCl3, ppm):
d 177.1, 174.7, 151.7, 138.8, 125.8, 125.8, 111.9, 111.9,
60.0, 58.2, 57.0, 56.5, 52.6, 49.5, 43.5, 42.3, 39.6, 38.1, 37.6, 35.1, 35.0,
4.3.6.11. Pyrazoline derivative 9k. Yield 73%; Mp 110.1e111.8 ꢀC; IR
(KBr): 3060, 2952, 2849, 1721, 1601, 1491, 1457, 1378, 1324, 1295,
1264, 1229, 1176, 1151, 1096, 1029, 992, 851, 773, 693 cmꢁ1; 1H NMR
29.6, 29.0, 21.7, 21.2, 19.6, 18.7, 14.1, 13.0; HRMS (ESI, m/z) calcd for
C
29H39N3O4Na [M þ Na]þ 516.2838. Found: 516.2845.
(400 MHz, CDCl3, ppm):
d
7.15 (t, J ¼ 8.0 Hz, 1H), 6.96 (s, 1H), 6.89
4.3.6.16. Pyrazoline derivative 9p. Yield 48%; Mp 112.8e114.2 ꢀC; IR
(KBr): 2955, 2850, 1721, 1610, 1588, 1493, 1454, 1379, 1347, 1318,
(dd, J ¼ 8.0, 1.8 Hz, 1H), 6.68 (d, J ¼ 8.0 Hz, 1H), 4.12 (q, J ¼ 7.2 Hz,
2H), 3.96 (t, J ¼ 8.6 Hz, 1H), 3.49e3.43 (m, 1H), 2.97 (dd, J ¼ 14.4,
8.6 Hz, 1H), 2.34 (s, 3H), 2.21e2.18 (m, 1H), 1.90e1.70 (m, 5H), 1.67e
1.53 (m, 4H), 1.46e1.33 (m, 2H),1.28 (t, J ¼ 7.2 Hz, 3H), 1.26e1.21 (m,
2H), 1.19 (s, 6H), 1.15e0.89 (m, 4H), 0.82 (s, 3H); 13C NMR (100 MHz,
1281, 1255, 1230, 1163, 1123, 1067, 1029, 996, 853, 785, 698 cmꢁ1
;
1H NMR (400 MHz, CDCl3, ppm):
d
7.34 (t, J ¼ 8.0 Hz, 1H), 7.28e7.23
(m, 2H), 7.07 (d, J ¼ 7.6 Hz,1H), 4.15e4.10 (m, 2H), 4.00 (t, J ¼ 9.0 Hz,
1H), 3.53e3.47 (m, 1H), 3.02 (dd, J ¼ 14.2, 8.6 Hz, 1H), 2.21e2.18 (m,
1H),1.92e1.70 (m, 5H), 1.67e1.53 (m, 4H),1.46e1.36 (m, 2H), 1.29 (t,
J ¼ 7.2 Hz, 3H), 1.25e1.21 (m, 2H), 1.19 (s, 6H), 1.16e0.90 (m, 4H),
CDCl3, ppm):
d 177.2, 171.8, 149.2, 138.7, 128.7, 120.5, 115.0, 111.4,
60.0, 58.6, 57.2, 56.7, 55.2, 49.6, 43.6, 42.0, 39.7, 39.7, 38.2, 37.8, 35.4,
35.2, 29.0, 21.9, 21.2, 21.4, 19.8,18.8, 14.2,13.1; HRMS (ESI, m/z) calcd
for C30H43N2O2 [M þ H]þ 463.3325. Found: 463.3327.
0.82 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2, 172.6, 149.1,
131.1 (q, J ¼ 31.0 Hz), 129.3, 124.3 (q, J ¼ 270.8 Hz), 117.1, 115.7 (q,
J ¼ 4.4 Hz), 110.4 (q, J ¼ 3.9 Hz), 60.0, 58.5, 57.1, 56.7, 54.6, 49.7, 43.6,
42.1, 39.7, 39.7, 38.2, 37.8, 35.4, 35.2, 29.0, 21.8, 21.3, 19.7, 18.8, 14.2,
13.1; HRMS (ESI, m/z) calcd for C30H40F3N2O2 [M þ H]þ 517.3042.
Found: 517.3040.
4.3.6.12. Pyrazoline derivative 9l. Yield 81%; Mp 74.5e76.3 ꢀC; IR
(KBr): 3095, 2950, 2848, 1720, 1616, 1513, 1456, 1377, 1322, 1234,
1179, 1150, 1095, 1030, 976, 852, 806, 516 cmꢁ1; 1H NMR (400 MHz,
CDCl3, ppm):
d
7.08 (d, J ¼ 8.4 Hz, 2H), 7.02 (d, J ¼ 8.4 Hz, 2H), 4.12
(q, J ¼ 7.2 Hz, 2H), 3.95 (t, J ¼ 8.8 Hz, 1H), 3.49e3.43 (m, 1H), 2.92
(dd, J ¼ 14.4, 8.6 Hz, 1H), 2.28 (s, 3H), 2.21e2.17 (m, 1H), 1.89e1.66
(m, 5H),1.64e1.52 (m, 4H), 1.45e1.31 (m, 2H), 1.28 (t, J ¼ 7.2 Hz, 3H),
1.25e1.20 (m, 2H), 1.19 (s, 3H), 1.18 (s, 3H), 1.15e0.86 (m, 4H), 0.81
4.3.6.17. Pyrazoline derivative 9q. Yield 65%; Mp 104.7e106.2 ꢀC; IR
(KBr): 3074, 2955, 2849, 1721, 1637, 1595, 1503, 1453, 1380, 1322,
1256, 1223, 1177, 1142, 1026, 977, 948, 846, 809, 598 cmꢁ1; 1H NMR
(400 MHz, CDCl3, ppm):
d 7.54e7.48 (m, 1H), 6.85e6.77 (m, 2H),
(s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d
177.2, 171.8, 147.3, 129.4,
4.17e4.09 (m, 3H), 3.53e3.48 (m, 1H), 2.88 (dd, J ¼ 14.4, 8.6 Hz, 1H),
2.21e2.17 (m, 1H), 1.88e1.71 (m, 5H), 1.68e1.53 (m, 4H), 1.45e1.30
(m, 3H), 1.27 (t, J ¼ 7.2 Hz, 3H), 1.25e1.24 (m, 1H), 1.22 (s, 3H),1.17 (s,
3H), 1.14e0.88 (m, 4H), 0.83 (s, 3H); 13C NMR (100 MHz, CDCl3,
129.4, 129.0, 114.5, 114.5, 60.0, 58.6, 57.2, 56.7, 55.7, 49.7, 43.6, 42.0,
39.8, 39.7, 38.2, 37.8, 35.4, 35.3, 29.0, 21.9, 21.4, 20.5, 19.8, 18.8, 14.2,
13.1; HRMS (ESI, m/z) calcd for C30H43N2O2 [M þ H]þ 463.3325.
Found: 463.3320.
ppm):
d
177.2, 173.6 (m), 157.8 (dd, J ¼ 242.5, 11.7 Hz), 152.7 (dd,
J ¼ 246.4, 12.4 Hz), 134.5 (dd, J ¼ 9.1, 3.2 Hz), 120.5 (dd, J ¼ 7.5,
3.5 Hz), 110.8 (dd, J ¼ 21.2, 3.4 Hz), 104.3 (t, J ¼ 25.6 Hz), 60.0, 59.5
(d, J ¼ 5.5 Hz), 58.6, 57.1, 56.7, 50.6, 43.6, 42.1, 39.7, 39.7, 38.2, 37.8,
35.2, 35.2, 29.0, 21.8, 21.4, 19.7,18.8, 14.1, 13.2; HRMS (ESI, m/z) calcd
for C29H39F2N2O2 [M þ H]þ 485.2980. Found: 485.2982.
4.3.6.13. Pyrazoline derivative 9m. Yield 51%; Mp 59.5e61.3 ꢀC; IR
(KBr): 3043, 2950, 2847, 1719, 1626, 1508, 1459, 1379, 1323, 1287,
1239, 1177, 1150, 1096, 1033, 975, 851, 823, 526 cmꢁ1 1H NMR
;
(400 MHz, CDCl3, ppm):
d 7.08e7.06 (m, 2H), 6.86e6.84 (m, 2H),
4.11 (q, J ¼ 7.2 Hz, 2H), 3.92 (t, J ¼ 8.6 Hz,1H), 3.77 (s, 3H), 3.49e3.43
(m, 1H), 2.89 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.21e2.17 (m, 1H), 1.89e1.73
(m, 5H), 1.71e1.53 (m, 4H), 1.45e1.35 (m, 2H), 1.28 (t, J ¼ 7.2 Hz, 3H),
1.25e1.20 (m, 2H), 1.19 (s, 3H), 1.18 (s, 3H), 1.15e0.89 (m, 4H), 0.82
4.3.6.18. Pyrazoline derivative 9r. Yield 42%; Mp 115.7e116.3 ꢀC; IR
(KBr): 2955, 2850, 1721, 1624, 1587, 1502, 1459, 1380, 1323, 1294,
1235, 1180, 1150, 1097, 1027, 982, 853, 971, 761, 607 cmꢁ1; 1H NMR
(s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d
177.2, 172.0, 153.7, 143.9,
(400 MHz, CDCl3, ppm): d 7.33e7.26 (m, 1H), 6.99e6.92 (m, 1H),
115.9, 115.9, 114.4, 114.4, 60.0, 58.6, 57.2, 56.7, 56.6, 55.6, 50.0, 43.6,
41.9, 39.8, 39.7, 38.2, 37.8, 35.4, 35.3, 29.0, 21.9, 21.4, 19.8, 18.8, 14.1,
13.1; HRMS (ESI, m/z) calcd for C30H43N2O3 [M þ H]þ 479.3274.
Found: 479.3269.
6.56e6.50 (m, 1H), 4.31e4.25 (m, 1H), 4.12 (q, J ¼ 7.2 Hz, 2H), 3.53e
3.47 (m, 1H), 2.99 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.21e2.17 (m, 1H), 1.87e
1.70 (m, 5H),1.68e1.52 (m, 4H),1.45e1.34 (m, 2H),1.28 (t, J ¼ 7.2 Hz,
3H), 1.25e1.22 (m, 2H), 1.22 (s, 3H), 1.18 (s, 3H), 1.14e0.85 (m, 4H),
0.82 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2, 173.3, 159.0
4.3.6.14. Pyrazoline derivative 9n. Yield 38%; Mp 185.5e187.3 ꢀC; IR
(d, J ¼ 237.2 Hz), 148.0 (d, J ¼ 237.0 Hz), 138.7 (t, J ¼ 10.5 Hz), 116.5
(dd, J ¼ 23.4, 10.0 Hz), 107.0 (dd, J ¼ 24.3, 7.8 Hz), 106.3 (dd, J ¼ 28.3,
3.7 Hz), 60.1, 58.5, 58.3 (d, J ¼ 7.9 Hz), 57.1, 56.7, 50.4, 43.6, 42.1, 39.8,
39.7, 38.2, 37.8, 35.2, 35.1, 29.0, 21.8, 21.4, 19.7, 18.8, 14.1, 13.2; HRMS
(ESI, m/z) calcd for C29H39F2N2O2 [M þ H]þ 485.2980. Found:
485.2978.
(KBr): 3095, 2955, 2845, 1716, 1616, 1568, 1525, 1486, 1461, 1381,
1340, 1255, 1226, 1149, 1097, 1025, 850, 733 cmꢁ1 1H NMR
;
(400 MHz, CDCl3, ppm):
d
7.81 (t, J ¼ 2.0 Hz, 1H), 7.67e7.65 (m, 1H),
7.43e7.40 (m, 1H), 7.37 (t, J ¼ 8.0 Hz, 1H), 4.16e4.10 (m, 2H), 4.02 (t,
J ¼ 8.6 Hz, 1H), 3.55e3.49 (m, 1H), 3.73 (dd, J ¼ 14.4, 8.6 Hz, 1H),

S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
77
4.3.6.19. Pyrazoline derivative 9s. Yield 41%; Mp 182.4e183.6 ꢀC; IR
(KBr): 3063, 2953, 2848, 1720, 1633, 1577, 1450, 1419, 1379, 1323,
1290, 1255, 1230, 1177, 1150, 1095, 1027, 989, 849, 779, 737, 710,
1.90e1.73 (m, 5H), 1.64e1.54 (m, 4H), 1.45e1.35 (m, 2H), 1.28 (t,
J ¼ 7.2 Hz, 3H), 1.26e1.21 (m, 2H), 1.19 (s, 3H), 1.18 (s, 3H), 1.15e0.93
(m, 4H), 0.80 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2,
642 cmꢁ1
;
1H NMR (400 MHz, CDCl3, ppm):
d
7.44 (dd, J ¼ 8.0,
172.9, 148.3, 132.5, 130.3, 121.9, 115.5, 113.6, 60.0, 58.5, 57.1, 56.6,
53.4, 49.8, 43.6, 42.1, 39.7, 39.7, 38.2, 37.8, 35.3, 35.2, 29.0, 21.8, 21.3,
19.7, 18.8, 14.2, 13.1; HRMS (ESI, m/z) calcd for C29H39Cl2N2O2
[M þ H]þ 517.2389. Found: 517.2387; [Mþ2 þ H]þ 519.2389. Found:
519.2375.
2.0 Hz, 1H), 7.19 (dd, J ¼ 8.0, 2.0 Hz, 1H), 7.15 (t, J ¼ 8.0 Hz, 1H), 4.34
(t, J ¼ 8.6 Hz, 1H), 4.18e4.05 (m, 2H), 3.57e3.51 (m, 1H), 2.73 (dd,
J ¼ 14.4, 8.6 Hz, 1H), 2.21e2.17 (m, 1H), 1.86e1.74 (m, 5H), 1.64e1.52
(m, 4H), 1.45e1.32 (m, 3H), 1.27 (t, J ¼ 7.2 Hz, 3H), 1.24 (s, 3H), 1.23e
1.19 (m, 1H), 1.17 (s, 3H), 1.13e0.89 (m, 4H), 0.84 (s, 3H); 13C NMR
(100 MHz, CDCl3, ppm):
d
177.2, 174.3, 149.5, 133.6, 127.4, 125.2,
4.3.6.24. Pyrazoline derivative 9x. Yield 59%; Mp 211.4e213.6 ꢀC; IR
124.8, 120.1, 60.1, 58.7, 57.1, 56.7, 53.4, 50.5, 43.6, 42.2, 39.8, 39.8,
38.2, 37.8, 35.2, 35.1, 28.9, 21.7, 21.5, 19.7, 18.8, 14.1, 13.3; HRMS (ESI,
m/z) calcd for C29H39Cl2N2O2 [M þ H]þ 517.2389. Found: 517.2385;
[Mþ2 þ H]þ 519.2389. Found: 519.2378.
(KBr): 3085, 2927, 2849, 1721, 1586, 1557, 1457, 1380, 1321, 1231,
1177, 1150, 1096, 1024, 983, 852, 822, 794, 670 cmꢁ1 1H NMR
;
(400 MHz, CDCl3, ppm):
d
6.94 (d, J ¼ 1.8 Hz, 2H), 6.79 (t, J ¼ 1.8 Hz,
1H), 4.15e4.08 (m, 2H), 3.91 (t, J ¼ 8.6 Hz, 1H), 3.50e3.44 (m, 1H),
3.01 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.21e2.18 (m, 1H), 1.90e1.73 (m, 5H),
1.64e1.51 (m, 4H), 1.46e1.34 (m, 2H), 1.29 (t, J ¼ 7.2 Hz, 3H), 1.25e
1.21 (m, 2H), 1.19 (s, 3H), 1.18 (s, 3H), 1.15e0.89 (m, 4H), 0.80 (s, 3H);
4.3.6.20. Pyrazoline derivative 9t. Yield 43%; Mp 102.7e104.1 ꢀC; IR
(KBr): 3096, 2952, 2848, 1723, 1635, 1584, 1555, 1473, 1328, 1322,
1261, 1228, 1173, 1148, 1098, 1054, 1027, 978, 853, 817, 774,
13C NMR (100 MHz, CDCl3, ppm):
d 177.1, 173.0, 150.1, 135.1, 135.1,
561 cmꢁ1; 1H NMR (400 MHz, CDCl3, ppm):
d
7.46 (d, J ¼ 8.4 Hz,1H),
118.7, 112.2, 112.2, 60.0, 58.4, 57.1, 56.6, 54.3, 49.7, 43.6, 42.1, 39.7,
39.7, 38.2, 37.8, 35.3, 35.2, 29.0, 21.8, 21.3, 19.7, 18.7, 14.2,13.1; HRMS
(ESI, m/z) calcd for C29H39Cl2N2O2 [M þ H]þ 517.2389. Found:
517.2386; [Mþ2 þ H]þ 519.2389. Found: 519.2414.
7.36 (d, J ¼ 2.4 Hz, 1H), 7.18 (dd, J ¼ 8.4, 2.4 Hz, 1H), 4.31 (t,
J ¼ 8.6 Hz, 1H), 4.16e4.07 (m, 2H), 3.56e3.50 (m, 1H), 2.70 (dd,
J ¼ 14.4, 8.6 Hz, 1H), 2.21e2.17 (m, 1H), 1.87e1.67 (m, 5H), 1.64e1.52
(m, 4H), 1.44e1.32 (m, 3H), 1.27 (t, J ¼ 7.2 Hz, 3H), 1.24 (s, 3H), 1.21e
1.20 (m, 1H), 1.17 (s, 3H), 1.13e0.86 (m, 4H), 0.83 (s, 3H); 13C NMR
4.3.6.25. Pyrazoline derivative 9y. Yield 79%; Mp 138.2e139.8 ꢀC; IR
(100 MHz, CDCl3, ppm):
d
177.2, 174.3, 146.4, 129.9, 128.8, 127.5,
(KBr): 3054, 2952, 2848, 1718, 1629, 1582, 1496, 1378, 1320, 1255,
126.5, 122.8, 60.3, 60.0, 58.7, 57.1, 56.7, 50.6, 43.6, 42.2, 39.8, 39.7,
38.2, 37.8, 35.2, 35.1, 29.0, 21.7, 21.5, 19.7, 18.8, 14.0, 13.3; HRMS (ESI,
m/z) calcd for C29H39Cl2N2O2 [M þ H]þ 517.2389. Found: 517.2390;
[Mþ2 þ H]þ 519.2389. Found: 519.2380.
1230, 1178, 1150, 1096, 1025, 978, 851, 778, 717 cmꢁ1 1H NMR
;
(400 MHz, CDCl3, ppm):
d
7.12 (d, J ¼ 7.2 Hz, 1H), 7.06 (t, J ¼ 7.2 Hz,
1H), 6.92 (d, J ¼ 7.2 Hz, 1H), 4.14e4.06 (m, 2H), 3.74 (t, J ¼ 8.6 Hz,
1H), 3.49e3.44 (m, 1H), 2.75 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.32 (s, 3H),
2.28 (s, 3H), 2.20e2.17 (m, 1H), 1.87e1.73 (m, 5H), 1.67e1.53 (m,
4H), 1.46e1.29 (m, 3H), 1.26 (t, J ¼ 7.2 Hz, 3H), 1.22 (s, 3H), 1.20e1.18
(m, 1H), 1.17 (s, 3H), 1.16e0.86 (m, 4H), 0.84 (s, 3H); 13C NMR
4.3.6.21. Pyrazoline derivative 9u. Yield 48%; Mp 154.9e156.3 ꢀC; IR
(KBr): 3094, 2945, 2844, 1722, 1637, 1580, 1509, 1460, 1393, 1320,
1248, 1227, 1171, 1150, 1093, 1049, 1012, 976, 879, 799, 605 cmꢁ1; 1H
(100 MHz, CDCl3, ppm):
d 177.3, 173.0, 149.2, 137.6, 130.1, 125.8,
NMR (400 MHz, CDCl3, ppm):
d
7.52 (d, J ¼ 2.5 Hz, 1H), 7.26 (d,
125.7, 117.5, 60.9, 60.0, 58.9, 57.2, 56.8, 50.4, 43.6, 42.0, 39.8, 39.8,
38.2, 37.8, 35.3, 35.3, 29.0, 21.8, 21.6, 20.5, 19.8, 18.8, 14.6, 14.1, 13.3;
HRMS (ESI, m/z) calcd for C31H45N2O2 [M þ H]þ 477.3481. Found:
477.3480.
J ¼ 2.5 Hz, 1H), 6.96 (dd, J ¼ 8.5, 2.5 Hz, 1H), 4.37 (t, J ¼ 8.8 Hz, 1H),
4.16e4.05 (m, 2H), 3.56e3.50 (m, 1H), 2.74 (dd, J ¼ 14.4, 8.6 Hz, 1H),
2.21e2.17 (m, 1H), 1.86e1.73 (m, 5H), 1.69e1.53 (m, 4H), 1.46e1.29
(m, 2H), 1.27 (t, J ¼ 7.2 Hz, 3H), 1.25 (s, 3H), 1.24e1.19 (m, 2H), 1.17 (s,
3H), 1.14e0.87 (m, 4H), 0.83 (s, 3H); 13C NMR (100 MHz, CDCl3,
4.3.6.26. Pyrazoline derivative 9z. Yield 75%; Mp 152.2e153.8 ꢀC; IR
ppm):
d
177.2, 174.5,148.4,133.0,131.1,124.0,123.9,121.9, 60.0, 59.9,
(KBr): 2952, 2848, 1721, 1630, 1502, 1452, 1378, 1320, 1255, 1229,
58.6, 57.1, 56.7, 50.5, 43.6, 42.2, 39.8, 39.7, 38.2, 37.8, 35.2, 35.1, 29.0,
1177, 1150, 1095, 1027, 977, 850, 813 cmꢁ1 1H NMR (400 MHz,
;
21.7, 21.5, 19.7, 18.8, 14.0, 13.2; HRMS (ESI, m/z) calcd for
C
519.2389. Found: 519.2382.
CDCl3, ppm):
d
7.10 (d, J ¼ 8.0 Hz,1H), 6.98 (s,1H), 6.96 (d, J ¼ 8.0 Hz,
29H39Cl2N2O2 [M þ H]þ 517.2389. Found: 517.2391; [Mþ2 þ H]þ
1H), 4.13e4.06 (m, 2H), 3.75 (t, J ¼ 8.6 Hz, 1H), 3.47e3.41 (m, 1H),
2.77 (dd, J ¼ 14.4, 8.6 Hz,1H), 2.37 (s, 3H), 2.27 (s, 3H), 2.20e2.16 (m,
1H), 1.85e1.72 (m, 5H), 1.67e1.54 (m, 4H), 1.44e1.28 (m, 3H), 1.25 (t,
J ¼ 7.2 Hz, 3H), 1.24e1.22 (m, 1H), 1.20 (s, 3H), 1.17 (s, 3H), 1.16e0.89
4.3.6.22. Pyrazoline derivative 9v. Yield 40%; Mp 154.9e156.3 ꢀC; IR
(KBr): 3060, 2924, 2848, 1720, 1628, 1558, 1446, 1377, 1322, 1229,
(m, 4H), 0.83 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2,
1178, 1150, 1095, 1026, 975, 848, 779, 699, 561 cmꢁ1
;
1H NMR
172.6, 146.7, 133.0, 131.6, 131.0, 126.9, 119.2, 60.3, 60.0, 58.9, 57.2,
56.8, 50.3, 43.6, 42.0, 39.8, 39.8, 38.2, 37.8, 35.3, 35.3, 29.0, 21.8, 21.5,
20.7, 19.8,18.9, 18.8, 14.1, 13.3; HRMS (ESI, m/z) calcd for C31H45N2O2
[M þ H]þ 477.3481. Found: 477.3482.
(400 MHz, CDCl3, ppm):
d
7.31 (d, J ¼ 8.0 Hz, 2H), 7.08 (t, J ¼ 8.0 Hz,
1H), 4.10 (q, J ¼ 7.2 Hz, 2H), 3.81 (dd, J ¼ 9.2, 8.0 Hz, 1H), 3.56e3.51
(m, 1H), 3.39 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.20e2.17 (m, 1H), 1.87e1.71
(m, 5H), 1.67e1.51 (m, 4H), 1.45e1.28 (m, 2H), 1.25 (s, 3H), 1.25 (t,
J ¼ 7.2 Hz, 3H), 1.22e1.18 (m, 2H), 1.17 (s, 3H), 1.15e0.85 (m, 4H),
4.3.6.27. Pyrazoline derivative 9aa. Yield 62%; Mp 62.2e64.3 ꢀC; IR
0.82 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d
177.2, 169.9, 142.1,
(KBr): 2952, 2848, 1721, 1069, 1557, 1504, 1454, 1411, 1378, 1321,
135.3,135.3,129.3,129.3,127.8, 60.0, 58.3, 57.4, 57.2, 56.8, 50.4, 43.6,
41.8, 40.2, 39.8, 38.2, 37.8, 35.4, 35.3, 29.0, 21.7, 21.1, 19.7, 18.8, 14.1,
13.2; HRMS (ESI, m/z) calcd for C29H39Cl2N2O2 [M þ H]þ 517.2389.
Found: 517.2386; [Mþ2 þ H]þ 519.2389. Found: 519.2379.
1257, 1230, 1178, 1151, 1095, 1028, 980, 849, 803 cmꢁ1 1H NMR
;
(400 MHz, CDCl3, ppm):
d
7.04 (d, J ¼ 8.0 Hz, 1H), 7.00 (s, 1H), 6.80
(d, J ¼ 8.0 Hz, 1H), 4.14e4.04 (m, 2H), 3.77 (t, J ¼ 8.6 Hz, 1H), 3.48e
3.42 (m, 1H), 2.81 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.36 (s, 3H), 2.30 (s, 3H),
2.20e2.17 (m, 1H), 1.85e1.70 (m, 5H), 1.65e1.55 (m, 4H), 1.46e1.29
(m, 3H),1.26 (t, J ¼ 7.2 Hz, 3H), 1.22 (s, 3H),1.22e1.20 (m, 1H),1.17 (s,
3H), 1.14e0.86 (m, 4H), 0.83 (s, 3H); 13C NMR (100 MHz, CDCl3,
4.3.6.23. Pyrazoline derivative 9w. Yield 58%; Mp 89.8e91.3 ꢀC; IR
(KBr): 2925, 2849, 1720, 1591, 1554, 1479, 1379, 1322, 1294, 1230,
1150, 1095, 1025, 990, 853, 802, 528 cmꢁ1
;
1H NMR (400 MHz,
ppm): d 177.2,172.7, 148.7, 135.9, 130.8,127.7,124.2, 119.5, 60.0, 59.9,
CDCl3, ppm):
d
7.27 (d, J ¼ 8.8 Hz, 1H), 7.17 (d, J ¼ 2.4 Hz, 1H), 6.90
58.8, 57.2, 56.8, 50.2, 43.6, 42.0, 39.8, 39.8, 38.2, 37.8, 35.3, 35.3,
29.0, 21.8, 21.5, 21.2, 19.8, 18.8, 18.8, 14.1, 13.3; HRMS (ESI, m/z) calcd
for C31H45N2O2 [M þ H]þ 477.3481. Found: 477.3479.
(dd, J ¼ 8.8, 2.4 Hz, 1H), 4.15e4.09 (m, 2H), 3.92 (t, J ¼ 8.6 Hz, 1H),
3.50e3.45 (m, 1H), 2.97 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.21e2.18 (m, 1H),

78
S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
4.3.6.28. Pyrazoline derivative 9ab. Yield 41%; Mp 57.6e58.3 ꢀC; IR
organic layer was washed with saturated NaCl aqueous solution,
dried with Na2SO4 and concentrated under vacuum. The residue
was purified by column chromatography on silica to give product
11ae11ad.
(KBr): 2953, 2848, 1721, 1621, 1467, 1378, 1322, 1231, 1178, 1151,
1097, 1027, 979, 851, 770 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
;
d
7.00 (s, 3H), 4.12e4.04 (m, 2H), 3.66 (t, J ¼ 8.6 Hz, 1H), 3.47e3.41
(m, 1H), 3.18 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.34 (s, 6H), 2.18e2.17 (m,
1H),1.87e1.66 (m, 5H),1.63e1.56 (m, 4H),1.54e1.28 (m, 3H), 1.24 (t,
J ¼ 7.2 Hz, 3H), 1.23 (s, 3H), 1.22e1.18 (m, 1H), 1.17 (s, 3H), 1.15e0.87
4.3.8.1. Pyrazole derivative 11a. Yield 76%; Mp 135.7e136.9 ꢀC; IR
(KBr): 3093, 3047, 2927, 2848, 1718, 1600, 1530, 1508, 1450, 1418,
1382, 1362, 1320, 1235, 1181, 1150, 1096, 1060, 1022, 985, 857, 767,
(m, 4H), 0.83 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.3,
167.6, 144.8, 136.4, 136.4, 129.0, 129.0, 126.2, 60.0, 59.2, 58.3, 57.2,
56.8, 50.0, 43.6, 41.6, 40.1, 39.8, 38.2, 37.9, 35.6, 35.4, 29.0, 21.7, 21.4,
19.8, 19.6, 19.6, 18.8, 14.0, 13.2; HRMS (ESI, m/z) calcd for
700, 648 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
; d 7.44e7.29 (m,
6H), 4.23e4.05 (m, 2H), 2.21e2.17 (m, 2H), 2.08e1.94 (m, 3H),1.87e
1.34 (m, 7H), 1.32e1.29 (t, J ¼ 7.2 Hz, 3H), 1.27e1.25 (m, 1H), 1.22 (s,
3H), 1.20e1.14 (m, 2H), 1.05 (s, 3H), 1.04e0.82 (m, 3H), 0.60 (s, 3H);
C
31H45N2O2 [M þ H]þ 477.3481. Found: 477.3484.
13C NMR (400 MHz, CDCl3, ppm):
d 177.4, 151.2, 139.9, 134.8, 131.5,
4.3.6.29. Pyrazoline derivative 9ac. Yield 73%; Mp 75.6e76.8 ꢀC; IR
128.8, 128.8, 127.1, 123.5, 123.5, 68.0, 60.0, 57.1, 52.5, 43.8, 42.5, 41.5,
40.3, 38.2, 38.1, 37.8, 35.8, 29.7, 29.0, 22.6, 21.4, 19.0, 14.2, 12.7;
HRMS (ESI, m/z) calcd for C29H39N2O2 [M þ H]þ 447.3012. Found:
447.3011.
(KBr): 2927, 2849, 1721, 1611, 1571, 1504, 1452, 1379, 1326, 1258,
1228, 1177, 1150, 1095, 1026, 850, 810 cmꢁ1 1H NMR (400 MHz,
;
CDCl3, ppm):
d
7.02 (d, J ¼ 8.0 Hz, 1H), 6.96 (d, J ¼ 2.4 Hz, 1H), 6.84
(dd, J ¼ 8.0, 2.4 Hz, 1H), 4.12 (q, J ¼ 7.2 Hz, 2H), 3.94 (t, J ¼ 8.6 Hz,
1H), 3.48e3.42 (m, 1H), 2.91 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.25 (s, 3H),
2.21e2.19 (m,1H), 2.19 (s, 3H),1.89e1.70 (m, 5H),1.67e1.52 (m, 4H),
1.45e1.32 (m, 3H), 1.28 (t, J ¼ 7.2 Hz, 3H), 1.25e1.20 (m, 1H), 1.19 (s,
3H), 1.18 (s, 3H), 1.17e0.89 (m, 4H), 0.82 (s, 3H); 13C NMR (100 MHz,
4.3.8.2. Pyrazole derivative 11b. Yield 87%; Mp 146.7e147.9 ꢀC; IR
(KBr): 3093, 2924, 2851, 1719, 1617, 1595, 1513, 1462, 1413, 1377,
1310, 1223, 1179, 1149, 1094, 1025, 989, 860, 818, 770, 689, 647,
510 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
; d 7.47e7.45 (m, 1H),
CDCl3, ppm):
d
177.2, 171.7, 147.7, 137.0, 129.9, 127.8, 116.0, 111.9,
7.43 (s, 1H), 7.39e7.33 (m, 1H), 7.23e7.16 (m, 2H), 4.20e4.06 (m,
2H), 2.21e2.15 (m, 2H), 2.08e1.94 (m, 3H), 1.84e1.35 (m, 7H), 1.32e
1.29 (t, J ¼ 7.2 Hz, 3H), 1.27e1.25 (m, 1H), 1.22 (s, 3H), 1.21e1.16 (m,
2H), 1.06e0.95 (m, 2H), 0.93 (s, 3H), 0.92e0.87 (m, 1H), 0.61 (s, 3H);
60.0, 58.6, 57.2, 56.7, 55.8, 49.7, 43.6, 42.0, 39.8, 39.7, 38.2, 37.8, 35.4,
35.3, 29.0, 21.9, 21.4, 20.1, 19.8, 18.8, 18.8, 14.2, 13.2; HRMS (ESI, m/z)
calcd for C31H45N2O2 [M þ H]þ 477.3481. Found: 477.3479.
13C NMR (100 MHz, CDCl3, ppm):
d
177.4, 156.4 (d, J ¼ 249.4 Hz),
4.3.6.30. Pyrazoline derivative 9ad. Yield 46%; Mp 156.6e158.2 ꢀC;
153.5, 135.4, 130.5, 129.6 (d, J ¼ 7.6 Hz), 128.4, 128.1 (d, J ¼ 11.6 Hz),
124.4 (d, J ¼ 3.8 Hz), 116.2 (d, J ¼ 19.7 Hz), 67.8, 60.0, 57.1, 52.7, 43.8,
42.8, 41.6, 40.2, 38.1, 38.1, 37.8, 34.8 (d, J ¼ 3.5 Hz), 29.0, 22.6, 21.3,
21.1, 19.0, 14.2, 12.7; HRMS (ESI, m/z) calcd for C29H38FN2O2
[M þ H]þ 465.2917. Found: 465.2916.
IR (KBr): 2950, 2848, 1721, 1597, 1464, 1379, 1345, 1299, 1228, 1095,
1032, 993, 851, 824, 689 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
;
d
6.75 (s, 2H), 6.52 (s, 1H), 4.12 (q, J ¼ 7.2 Hz, 2H), 3.95 (t, J ¼ 8.6 Hz,
1H), 3.48e3.42 (m, 1H), 2.96 (dd, J ¼ 14.4, 8.6 Hz, 1H), 2.29 (s, 6H),
2.21e2.17 (m, 1H), 1.90e1.70 (m, 5H), 1.67e1.55 (m, 4H), 1.45e1.32
(m, 2H), 1.27 (t, J ¼ 7.2 Hz, 3H), 1.23e1.20 (m, 2H), 1.19 (s, 6H), 1.15e
4.3.8.3. Pyrazole derivative 11c. Yield 93%; Mp 147.0e147.9 ꢀC; IR
(KBr): 3095, 2942, 2863, 1710, 1611, 1532, 1502, 1456, 1403, 1378,
1360, 1239, 1180, 1148,1089,1032, 970, 871, 783, 689 cmꢁ1; 1H NMR
0.89 (m, 4H), 0.81 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.2,
171.6, 149.2, 138.5, 138.5, 121.6, 112.1, 112.1, 60.0, 58.6, 57.2, 56.7,
55.2, 49.6, 43.6, 42.0, 39.7, 39.7, 38.2, 37.8, 35.4, 35.2, 29.0, 21.9, 21.5,
21.5, 21.4,19.8,18.8,14.2,13.1; HRMS (ESI, m/z) calcd for C31H45N2O2
[M þ H]þ 477.3481. Found: 477.3484.
(400 MHz, CDCl3, ppm):
d 7.41 (s, 1H), 7.39e7.35 (m, 1H), 7.20e7.17
(m, 2H), 7.04e7.00 (m, 1H), 4.20e4.05 (m, 2H), 2.21e2.17 (m, 2H),
2.07e1.93 (m, 3H), 1.87e1.38 (m, 7H), 1.32e1.29 (t, J ¼ 7.2 Hz, 3H),
1.27e1.24 (m, 1H), 1.22 (s, 3H), 1.21e1.17 (m, 2H), 1.10 (s, 3H), 1.06e
4.3.7. Synthesis of compound 10
0.88 (m, 3H), 0.58 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.4,
Compound 3 (0.378 g, 1 mmol) was dissolved in CH2Cl2 (20 mL)
and then TCC/Silica gel (0.334 g) was added. After stirring at room
temperature for 1.5 h, the reaction mixture was filtered and washed
with CH2Cl2. At last the combined organic phases were washed
with saturated NaCl aqueous solution, dried with Na2SO4 and the
filtrate was concentrated. The residue was purified by column
chromatography on silica (petroleum ether/ethyl acetate ¼ 3:1, v/v)
to give product 10 (0.233 g, 62%). Mp 110.7e112.6 ꢀC; IR (KBr):
3439, 2955, 2924, 2850, 1719, 1668, 1602, 1457, 1392, 1342, 1233,
162.6 (d, J ¼ 245.4 Hz), 151.3, 141.3 (d, J ¼ 9.9 Hz), 135.3, 132.1, 130.0
(d, J ¼ 8.9 Hz), 118.9 (d, J ¼ 3.0 Hz), 113.9 (d, J ¼ 21.0 Hz), 110.9 (d,
J ¼ 24.5 Hz), 68.1, 60.0, 57.0, 52.5, 43.8, 42.5, 41.6, 40.2, 38.2, 38.1,
37.7, 35.7, 29.0, 22.6, 21.9, 21.3,19.0,14.2,12.6; HRMS (ESI, m/z) calcd
for C29H38FN2O2 [M þ H]þ 465.2917. Found: 465.2920.
4.3.8.4. Pyrazole derivative 11d. Yield 91%; Mp 118.3e120.1 ꢀC; IR
(KBr): 3086, 2944, 2847, 1717, 1626, 1518, 1450, 1385, 1217, 1151,
1096, 1025, 988, 850, 526 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
;
1207, 1183, 1156, 1093, 1038, 958, 830 cmꢁ1
CDCl3, ppm):
12.0 (d, J ¼ 1.1 Hz, 1H), 7.15 (s, 1H), 4.14e4.09 (q,
J ¼ 6.1 Hz, 2H), 2.18e1.37 (m, 14H), 1.30e1.26 (t, J ¼ 7.0 Hz, 3H), 1.21
(s, 3H), 1.18 (s, 2H), 1.02 (s, 3H), 0.98e0.74 (m, 2H), 0.67 (s, 3H); 13
NMR (100 MHz, CDCl3, ppm): 215.1, 177.2, 156.1, 121.3, 60.0, 56.5,
;
1H NMR (400 MHz,
d 7.40e7.35 (m, 3H), 7.14e7.08 (m, 2H), 4.20e4.07 (m, 2H), 2.22e
d
2.16 (m, 2H), 2.07e1.93 (m, 3H), 1.84e1.34 (m, 7H), 1.32e1.29 (t,
J ¼ 7.2 Hz, 3H), 1.27e1.25 (m, 1H), 1.22 (s, 3H), 1.20e1.17 (m, 2H),
1.03 (s, 3H), 1.03e0.83 (m, 3H), 0.59 (s, 3H); 13C NMR (100 MHz,
C
d
CDCl3, ppm):
d
177.4, 161.5 (d, J ¼ 245.3 Hz), 151.4, 136.1 (d,
56.3, 55.0, 47.3, 43.7, 41.3, 40.4, 38.3, 38.1, 38.0, 36.1, 28.9, 22.3, 20.9,
19.6, 18.9, 14.2, 12.3; HRMS (ESI, m/z) calcd for C23H34O4Na
[M þ Na]þ 397.2355. Found: 397.2354.
J ¼ 2.8 Hz), 134.8, 131.4, 125.4 (d, J ¼ 8.3 Hz), 125.4 (d, J ¼ 8.3 Hz),
115.7 (d, J ¼ 22.7 Hz), 115.7 (d, J ¼ 22.7 Hz), 67.9, 60.0, 57.1, 52.5,
43.8, 42.5, 41.5, 40.3, 38.2, 38.1, 37.7, 35.9, 29.0, 22.6, 21.8, 21.4, 19.0,
14.2, 12.6; HRMS (ESI, m/z) calcd for C29H38FN2O2 [M þ H]þ
465.2917. Found: 465.2915.
4.3.8. General procedure for synthesis of compounds 11ae11ad
A mixture of compound 10 (0.376 g, 1 mmol) and various phe-
nylhydrazines 6ae6ad (0.145e0.225 g, 1 mmol) in isopropanol
(10 mL) were stirred under reflux. After completion of the reaction
monitored by TLC, the reaction mixture was concentrated under
vacuum, and extracted with CH3CO2C2H5 and H2O. At last the
4.3.8.5. Pyrazole derivative 11e. Yield 87%; Mp 121.6e123.0 ꢀC; IR
(KBr): 3091, 2951, 2846, 1719, 1592, 1510, 1469, 1447, 1414, 1380,
1235, 1180, 1150, 1090, 1043, 985, 853, 764, 669 cmꢁ1 1H NMR
;
(400 MHz, CDCl3, ppm):
d
7.51e7.49 (d, J ¼ 7.8 Hz, 1H), 7.42 (s, 1H),

S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
79
7.40e7.31 (m, 3H), 4.22e4.06 (m, 2H), 2.20e2.17 (m, 2H), 2.08e1.97
(m, 3H), 1.85e1.36 (m, 8H), 1.33e1.29 (t, J ¼ 7.2 Hz, 3H), 1.23 (s, 3H),
1.21e1.15 (m, 2H), 1.06e0.86 (m, 3H), 0.84 (s, 3H), 0.63 (s, 3H); 13
NMR (100 MHz, CDCl3, ppm): 177.5, 153.5, 137.8, 135.0, 132.5,
130.2, 130.0, 129.5, 127.0, 127.0, 67.2, 60.0, 57.1, 52.6, 43.8, 43.0, 41.1,
40.3, 38.2, 38.1, 37.8, 35.1, 29.0, 22.6, 21.4, 21.1, 19.0,14.2,12.7; HRMS
(ESI, m/z) calcd for C29H38ClN2O2 [M þ H]þ 481.2622 Found:
481.2621; [Mþ2 þ H]þ 483.2622. Found: 483.2608.
1H NMR (400 MHz, CDCl3, ppm):
d
7.55 (d, J ¼ 8.8 Hz, 2H), 7.40 (s,
1H), 7.29 (d, J ¼ 8.8 Hz, 2H), 4.20e4.07 (m, 2H), 2.21e2.16 (m, 2H),
2.07e1.92 (m, 3H), 1.83e1.33 (m, 7H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.27e
1.23 (m, 1H), 1.22 (s, 3H), 1.20e1.16 (m, 2H), 1.06 (s, 3H), 1.05e0.87
C
d
(m, 3H), 0.58 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.4,151.3,
139.0, 135.2, 132.0, 131.9, 131.9, 124.9, 124.9, 120.5, 68.0, 60.0, 57.0,
52.5, 43.8, 42.5, 41.6, 40.2, 38.2, 38.1, 37.7, 35.8, 29.0, 22.6, 21.9, 21.3,
19.0, 14.2, 12.6; HRMS (ESI, m/z) calcd for C29H38BrN2O2 [M þ H]þ
525.2117. Found: 525.2119; [Mþ2 þ H]þ 527.2117. Found: 527.2114.
4.3.8.6. Pyrazole derivative 11f. Yield 76%; Mp 117.0e118.3 ꢀC; IR
(KBr): 3081, 2937, 2847, 1715, 1625, 1594, 1531, 1497, 1465, 1378,
1341, 1231, 1178, 1148, 1097, 1073, 1025, 996, 867, 779, 686,
4.3.8.11. Pyrazole derivative 11k. Yield 56%; Mp 115.2e116.7 ꢀC; IR
(KBr): 3088, 2940, 2870, 2847, 1714, 1636, 1584, 1512, 1465, 1446,
1416, 1377, 1320, 1225, 1170, 1150, 1094, 1016, 989, 854, 771, 722,
643 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
; d 7.45e7.45 (t,
J ¼ 2.0 Hz, 1H),7.41 (s, 1H), 7.37e7.33 (t, J ¼ 8.0 Hz, 1H), 7.31e7.26
(m, 2H), 4.20e4.07 (m, 2H), 2.21e2.17 (m, 2H), 2.07e1.92 (m, 3H),
1.86e1.38 (m, 8H), 1.32e1.29 (t, J ¼ 7.2 Hz, 3H), 1.22 (s, 3H), 1.21e
1.17 (m, 2H), 1.09 (s, 3H), 1.06e0.88 (m, 3H), 0.58 (s, 3H); 13C NMR
454 cmꢁ1; 1H NMR (400 MHz, CDCl3, ppm):
d 7.37 (s, 1H), 7.33e7.31
(m, 1H), 7.30e7.26 (m, 1H), 7.24e7.20 (m, 1H), 7.18e7.16 (m, 1H),
4.20e4.08 (m, 2H), 2.21e2.15 (m, 2H), 2.11 (s, 3H), 2.09e1.97 (m,
3H), 1.82e1.36 (m, 7H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.30e1.25 (m, 1H),
1.23 (s, 3H), 1.20e1.17 (m, 2H), 1.06e0.84 (m, 3H), 0.81 (s, 3H), 0.63
(100 MHz, CDCl3, ppm):
d 177.4, 151.3, 140.9, 135.4, 134.5, 132.1,
129.8, 127.0, 123.6, 121.4, 68.1, 60.0, 57.0, 52.5, 43.8, 42.5, 41.6, 40.2,
38.2, 38.1, 37.7, 35.7, 29.0, 22.6, 22.0, 21.4,19.0,14.2,12.6; HRMS (ESI,
m/z) calcd for C29H38ClN2O2 [M þ H]þ 481.2622. Found: 481.2620;
[Mþ2 þ H]þ 483.2622. Found: 483.2661.
(s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.5, 152.6, 139.0, 136.0,
134.2,130.7,129.1,128.8,127.4,125.9, 67.2, 60.0, 57.1, 52.7, 43.8, 42.9,
41.0,40.3, 38.2, 38.1, 37.9, 35.5, 29.0, 22.6, 21.6, 21.3, 19.0, 17.2, 14.2,
12.7; HRMS (ESI, m/z) calcd for C30H41N2O2 [M þ H]þ 461.3168.
Found: 461.3167.
4.3.8.7. Pyrazole derivative 11g. Yield 50%; Mp 171.4e172.3 ꢀC; IR
(KBr): 3098, 3065, 2955, 2845, 1719, 1597, 1502, 1464, 1446, 1418,
4.3.8.12. Pyrazole derivative 11l. Yield 87%; Mp 119.2e120.5 ꢀC; IR
(KBr): 3088, 2954, 2848, 1712, 1611, 1593, 1529, 1501, 1447, 1409,
1380, 1235, 1181, 1150, 1093, 1026, 855, 786, 694, 644 cmꢁ1; 1H NMR
1384, 1361, 1336, 1233, 1181, 1149, 1092, 1030, 984, 835, 523 cmꢁ1
;
1H NMR (400 MHz, CDCl3, ppm):
d 7.41e7.34 (m, 5H), 4.20e4.06 (m,
2H), 2.21e2.16 (m, 2H), 2.07e1.35 (m, 11H), 1.31 (t, J ¼ 7.2 Hz, 3H),
(400 MHz, CDCl3, ppm): d 7.39 (s, 1H), 7.31e7.27 (m, 2H), 7.17e7.12
1.22 (s, 3H), 1.20e1.16 (m, 2H), 1.06 (s, 3H), 1.05e0.95 (m, 3H), 0.58
(m, 2H), 4.23e4.05 (m, 2H), 2.39 (s, 3H), 2.21e2.16 (m, 2H), 2.08e
1.93 (m, 3H), 1.86e1.37 (m, 8H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.22 (s, 3H),
1.20e1.17 (m, 2H), 1.05 (s, 3H), 1.03e0.88 (m, 3H), 0.59 (s, 3H); 13C
(s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.4, 151.3, 138.4, 135.1,
132.6, 131.9, 128.9, 128.9, 124.6, 124.6, 68.0, 60.0, 57.0, 52.4, 43.8,
42.4, 41.5, 40.2, 38.1, 38.0, 37.7, 35.7, 28.9, 22.5, 21.9, 21.3, 18.9, 14.1,
12.6; HRMS (ESI, m/z) calcd for C29H38N2O2 [M þ H]þ 481.2622
Found: 481.2622; [Mþ2 þ H]þ 483.2622. Found: 483.2636.
NMR (100 MHz, CDCl3, ppm): d 177.4, 151.2, 139.8, 138.9, 134.6,131.4,
128.5, 127.9, 124.3, 120.6, 68.0, 60.0, 57.1, 52.6, 43.8, 42.5, 41.5, 40.3,
38.2, 38.1, 37.8, 35.9, 29.0, 22.6, 21.9, 21.4, 21.3, 19.0, 14.2, 12.7; HRMS
(ESI, m/z) calcd for C30H41N2O2 [M þ H]þ 461.3168. Found: 461.3169.
4.3.8.8. Pyrazole derivative 11h. Yield 55%; Mp 142.8e143.9 ꢀC; IR
(KBr): 3088, 3058, 2952, 2847, 1715, 1632, 1588, 1533, 1509, 1447,
1380, 1341, 1322, 1303, 1235, 1180, 1149, 1093, 1028, 986, 852, 766,
4.3.8.13. Pyrazole derivative 11m. Yield 89%; Mp 193.8e194.5 ꢀC; IR
(KBr): 3096, 3048, 2946, 2844, 1719, 1612, 1521, 1446, 1418, 1384,
655 cmꢁ1
;
1H NMR (400 MHz, CDCl3, ppm):
d
7.69e7.67 (d,
1360, 1337, 1233, 1182, 1150, 1091, 1030, 984, 825, 521 cmꢁ1 1H
;
J ¼ 7.6 Hz, 1H), 7.41 (s, 1H), 7.38e7.31 (m, 3H), 4.17e4.11 (m, 2H),
2.21e1.36 (m,13H),1.31 (t, J ¼ 7.2 Hz, 3H),1.25 (s, 3H),1.20e0.86 (m,
5H), 0.84 (s, 3H), 0.64 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
NMR (400 MHz, CDCl3, ppm):
d
7.38 (s, 1H), 7.27 (t, J ¼ 6.6 Hz, 2H),
7.21 (d, J ¼ 8.3 Hz, 2H), 4.21e4.06 (m, 2H), 2.38 (s, 3H), 2.21e2.19
(m, 1H), 2.16 (dd, J ¼ 9.9, 2.8 Hz, 1H), 2.04e1.37 (m, 11H), 1.31 (t,
J ¼ 7.2 Hz, 3H), 1.22 (s, 3H), 1.20e1.16 (m, 2H), 1.04 (s, 3H), 1.02e0.88
d
177.5, 153.2, 139.4, 134.9, 133.2, 130.5, 129.6, 129.4, 127.7, 122.6,
67.1, 60.0, 57.1, 52.5, 43.8, 43.0, 41.0, 40.2, 38.1, 38.1, 37.8, 35.1, 29.0,
(m, 3H), 0.60 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.4,151.2,
22.6, 21.4, 21.1, 19.0, 14.2, 12.7; HRMS (ESI, m/z) calcd for
C
527.2117. Found: 527.2106.
137.5, 136.9, 134.5, 131.1, 129.3, 129.3, 123.5, 123.5, 67.9, 59.9, 57.0,
52.5, 43.8, 42.5, 41.4, 40.2, 38.1, 38.1, 37.8, 35.8, 28.9, 22.6, 21.8, 21.3,
21.0, 19.0, 14.1, 12.6; HRMS (ESI, m/z) calcd for C30H41N2O2 [M þ H]þ
461.3168. Found: 461.3167.
29H38BrN2O2 [M þ H]þ 525.2117. Found: 525.2117; [Mþ2 þ H]þ
4.3.8.9. Pyrazole derivative 11i. Yield 64%; Mp 118.8e119.5 ꢀC; IR
(KBr): 3083, 2939, 2847, 1714, 1626, 1592, 1496, 1469, 1375, 1227,
4.3.8.14. Pyrazole derivative 11n. Yield 61%; Mp 185.9e187.5 ꢀC; IR
(KBr): 3095, 2958, 2931, 2869, 2849, 1726, 1609, 1536, 1519, 1448,
1364, 1225, 1149, 1094, 1029, 989, 853, 784, 758, 524 cmꢁ1; 1H NMR
1180, 1148, 1092, 1027, 994, 871, 789, 757, 690, 642 cmꢁ1 1H NMR
;
(400 MHz, CDCl3, ppm):
d
7.61 (t, J ¼ 1.9 Hz, 1H), 7.44 (ddd, J ¼ 8.0,
1.8, 1.2 Hz, 1H), 7.41 (s, 1H), 7.35 (ddd, J ¼ 8.0, 1.9, 1.2 Hz, 1H), 7.29 (t,
J ¼ 8.0 Hz, 1H), 4.20e4.07 (m, 2H), 2.21e2.17 (m, 2H), 2.07e1.92 (m,
3H), 1.86e1.36 (m, 7H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.27e1.23 (m, 1H),
1.22 (s, 3H), 1.21e1.17 (m, 2H), 1.09 (s, 3H), 1.06e0.88 (m, 3H), 0.58
(400 MHz, CDCl3, ppm):
d
7.94 (dd, J ¼ 8.0, 1.4 Hz, 1H), 7.66 (dt,
J ¼ 7.6, 1.4 Hz, 1H), 7.55 (dt, J ¼ 7.6, 1.4 Hz, 1H), 7.45 (dd, J ¼ 7.6,
1.4 Hz,1H), 7.39 (s, 1H), 4.17e4.07 (m, 2H), 2.25e2.18 (m, 2H), 2.06e
1.95 (m, 3H), 1.82e1.36 (m, 7H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.27e1.24 (m,
1H), 1.22 (s, 3H), 1.20e1.16 (m, 2H), 1.06e1.00 (m, 2H), 0.98 (s, 3H),
0.95e0.87 (m, 1H), 0.61 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
(s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.4, 151.3, 141.0, 135.4,
132.1, 130.0, 130.0, 126.5, 122.3, 121.8, 68.1, 60.0, 57.0, 52.5, 43.8,
42.5, 41.6, 40.2, 38.2, 38.1, 37.7, 35.7, 29.0, 22.6, 22.0, 21.4, 19.0, 14.2,
12.6; HRMS (ESI, m/z) calcd for C29H38BrN2O2 [M þ H]þ 525.2117.
Found: 525.2112; [Mþ2 þ H]þ 527.2117. Found: 527.2081.
d
177.4, 152.9, 146.0, 136.3, 133.5, 132.7, 131.1, 129.1, 128.8, 125.0,
67.4, 60.0, 57.0, 52.4, 43.8, 42.9, 41.4, 40.2, 38.1, 38.1, 37.7, 35.4, 29.0,
22.5, 21.9, 21.1, 19.0, 14.2, 12.6; HRMS (ESI, m/z) calcd for
C
29H38N3O4 [M þ H]þ 492.2862. Found: 492.2865.
4.3.8.10. Pyrazole derivative 11j. Yield 76%; Mp 174.8e175.5 ꢀC; IR
(KBr): 3095, 2950, 2929, 2845, 1719, 1626, 1592, 1528, 1499, 1463,
4.3.8.15. Pyrazole derivative 11o. Yield 89%; Mp 138.9e140.7 ꢀC; IR
(KBr): 3108, 3085, 2951, 2899, 2841, 1723, 1618, 1590, 1537, 1494,
1446, 1416, 1396, 1232, 1180, 1150, 1091, 1029, 984, 833, 520 cmꢁ1
;

80
S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
1444, 1401, 1353, 1232, 1177, 1145, 1085, 1027, 998, 895, 858, 753,
679 cmꢁ1; 1H NMR (400 MHz, CDCl3, ppm):
29.0, 22.6, 21.1, 19.0, 14.2, 12.7; HRMS (ESI, m/z) calcd for
C
517.2232. Found: 517.2222.
d
8.25 (t, J ¼ 2.0 Hz,1H),
29H37Cl2N2O2 [M þ H]þ 515.2232. Found: 515.2228; [Mþ2 þ H]þ
8.17 (dd, J ¼ 8.0, 2.0 Hz, 1H), 7.86 (dd, J ¼ 8.0, 1.0 Hz, 1H), 7.63 (t,
J ¼ 8.0 Hz, 1H), 7.46 (s, 1H), 4.22e4.06 (m, 2H), 2.24e2.17 (m, 2H),
2.07e1.90 (m, 4H), 1.83e1.38 (m, 7H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.27e
1.25 (m, 1H), 1.23 (s, 3H), 1.21e1.18 (m, 1H), 1.13 (s, 3H), 1.07e0.87
4.3.8.20. Pyrazole derivative 11t. Yield 90%; Mp 147.7e148.9 ꢀC; IR
(KBr): 3090, 3063, 2954, 2848, 1712, 1559, 1533, 1503, 1467, 1448,
1380, 1339, 1305, 1235, 1181, 1151, 1100, 1042, 985, 858, 834, 807,
(m, 3H), 0.58 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.4,151.5,
148.3, 140.8, 136.1, 133.0, 129.9, 128.5, 121.3, 117.8, 68.1, 60.0, 57.0,
52.4, 43.8, 42.5, 41.8, 40.2, 38.1, 38.1, 37.6, 35.6, 29.0, 22.5, 22.1, 21.3,
19.0, 14.2, 12.6; HRMS (ESI, m/z) calcd for C29H38N3O4 [M þ H]þ
492.2862. Found: 492.2861.
628, 511 cmꢁ1; 1H NMR (400 MHz, CDCl3, ppm):
d
7.52 (d, J ¼ 2.0 Hz,
1H), 7.41 (s, 1H), 7.33 (dd, J ¼ 8.4, 2.0 Hz, 1H), 7.30 (d, J ¼ 8.4 Hz, 1H),
4.21e4.06 (m, 2H), 2.21e2.17 (m, 2H), 2.07e1.95 (m, 3H), 1.81e1.36
(m, 8H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.22 (s, 3H), 1.21e1.16 (m, 2H), 1.06e
0.90 (m, 3H), 0.86 (s, 3H), 0.62 (s, 3H); 13C NMR (100 MHz, CDCl3,
4.3.8.16. Pyrazole derivative 11p. Yield 57%; Mp 196.3e198.1 ꢀC; IR
(KBr): 3101, 3093, 2965, 2937, 2841, 1711, 1595, 1520, 1472, 1454,
1407, 1382, 1337, 1238, 1182, 1158, 1106, 1038, 982, 855, 752, 692,
ppm): d 177.4, 153.6, 136.5, 135.5, 135.4, 133.4, 130.2, 129.9, 129.9,
127.4, 67.2, 60.0, 57.1, 52.6, 43.8, 43.0, 41.2, 40.2, 38.1, 38.1, 37.8, 35.0,
29.0, 22.6, 21.5, 21.1, 19.0, 14.2, 12.7; HRMS (ESI, m/z) calcd for
640, 596, 515 cmꢁ1
;
1H NMR (400 MHz, CDCl3, ppm):
d
8.32 (d,
C
29H37Cl2N2O2 [M þ H]þ 515.2232. Found: 515.2233; [Mþ2 þ H]þ
J ¼ 8.9 Hz, 2H), 7.60 (d, J ¼ 8.9 Hz, 2H), 7.49 (s, 1H), 4.19e4.09 (m,
2H), 2.24e2.18 (m, 2H), 2.07e1.40 (m, 11H), 1.31 (t, J ¼ 7.1 Hz, 3H),
1.23 (s, 3H), 1.22e1.17 (m, 2H), 1.15 (s, 3H), 1.06e0.93 (m, 3H), 0.57
517.2232. Found: 517.2221.
4.3.8.21. Pyrazole derivative 11u. Yield 76%; Mp 192.9e194.3 ꢀC; IR
(KBr): 3086, 2955, 2848, 1719, 1585, 1511, 1467, 1448, 1383, 1338,
1232, 1181, 1149, 1092, 1043, 996, 852, 832, 799, 582 cmꢁ1; 1H NMR
(s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.3, 151.6, 145.7, 144.6,
136.6, 133.7, 124.6, 124.6, 122.7, 122.7, 68.3, 60.0, 56.9, 52.3, 43.7,
42.4, 41.9, 40.1, 38.1, 38.0, 37.4, 35.4, 28.9, 22.5, 22.1, 21.3, 18.9, 14.1,
12.6; HRMS (ESI, m/z) calcd for C29H38N3O4 [M þ H]þ 492.2862.
Found: 492.2862.
(400 MHz, CDCl3, ppm):
d
7.43 (d, J ¼ 8.4 Hz, 1H), 7.42 (s, 1H), 7.36
(dd, J ¼ 8.4, 2.4 Hz, 2H), 4.20e4.08 (m, 2H), 2.21e2.17 (m, 2H),
2.07e1.95 (m, 3H), 1.82e1.36 (m, 8H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.22 (s,
3H), 1.20e1.16 (m, 2H), 1.06e0.90 (m, 3H), 0.89 (s, 3H), 0.62 (s, 3H);
4.3.8.17. Pyrazole derivative 11q. Yield 87%; Mp 144.8e145.7 ꢀC; IR
13C NMR (100 MHz, CDCl3, ppm):
d 177.4, 153.6, 138.1, 135.5, 132.6,
(KBr): 3077, 2943, 2853, 1720, 1608, 1526, 1441, 1379, 1315, 1266,
130.9, 130.8, 130.2, 130.0, 129.7, 67.3, 60.0, 57.1, 52.6, 43.8, 43.0, 41.2,
40.2, 38.1, 38.1, 37.8, 35.0, 29.0, 22.6, 21.5, 21.1,19.0,14.2, 12.7; HRMS
(ESI, m/z) calcd for C29H37Cl2N2O2 [M þ H]þ 515.2232. Found:
515.2234; [Mþ2 þ H]þ 517.2232. Found: 517.2224.
1225, 1147, 1107, 1048, 1023, 959, 882, 830, 622 cmꢁ1 1H NMR
;
(400 MHz, CDCl3, ppm):
d 7.46e7.41 (m, 2H), 6.98e6.92 (m, 2H),
4.20e4.06 (m, 2H), 2.21e2.15 (m, 2H), 2.07e1.93 (m, 3H), 1.81e1.37
(m, 8H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.22 (s, 3H), 1.21e1.16 (m, 2H), 1.06e
0.96 (m, 1H), 0.93 (s, 3H), 0.91e0.87 (m, 2H), 0.60 (s, 3H); 13C NMR
4.3.8.22. Pyrazole derivative 11v. Yield 56%; Mp 181.5e182.6 ꢀC; IR
(KBr): 3086, 2983, 2949, 2847, 1721, 1631, 1565, 1512, 1463, 1436,
1378, 1340, 1234, 1181, 1152, 1093, 1042, 984, 855, 791, 751,
(100 MHz, CDCl3, ppm):
d
177.4, 162.1 (dd, J ¼ 249.4, 10.9 Hz), 156.7
(dd, J ¼ 252.0, 12.5 Hz), 153.6, 135.6, 130.6, 129.4 (d, J ¼ 10.2 Hz),
124.7 (dd, J ¼ 11.6, 4.0 Hz), 111.6 (dd, J ¼ 22.3, 3.9 Hz), 104.6 (dd,
J ¼ 26.2, 23.5 Hz), 67.7, 60.0, 57.1, 52.6, 43.8, 42.8, 41.5, 40.2, 38.1,
38.1, 37.8, 34.8 (d, J ¼ 3.2 Hz), 29.0, 22.6, 21.2, 21.1, 19.0, 14.1, 12.7;
HRMS (ESI, m/z) calcd for C29H37F2N2O2 [M þ H]þ 483.2823. Found:
483.2826.
531 cmꢁ1; 1H NMR (400 MHz, CDCl3, ppm):
d 7.48 (s, 1H), 7.44e7.41
(m, 2H), 7.33 (t, J ¼ 8.0 Hz, 1H), 4.20e4.08 (m, 2H), 2.22e2.18 (m,
2H), 2.09e1.98 (m, 3H), 1.86e1.34 (m, 7H), 1.31 (t, J ¼ 7.2 Hz, 3H),
1.27e1.25 (m, 1H), 1.23 (s, 3H), 1.21e1.17 (m, 2H), 1.06e0.89 (m, 3H),
0.88 (s, 3H), 0.64 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.5,
153.3, 136.0, 135.8, 135.6, 135.2, 130.6, 129.6, 128.4, 128.3, 67.0, 60.0,
57.1, 52.5, 43.8, 43.2, 41.0, 40.2, 38.1, 38.1, 37.8, 34.7, 29.0, 22.6, 20.9,
20.5, 19.0, 14.2, 12.7; HRMS (ESI, m/z) calcd for C29H37Cl2N2O2
[M þ H]þ 515.2232. Found: 515.2229; [Mþ2 þ H]þ 517.2232. Found:
517.2243.
4.3.8.18. Pyrazole derivative 11r. Yield 88%; Mp 177.8e178.3 ꢀC; IR
(KBr): 3086, 2927, 2854, 1717, 1626, 1515, 1443, 1384, 1333, 1242,
1180, 1151, 1107, 1039, 837, 813, 770, 644, 575 cmꢁ1 1H NMR
;
(400 MHz, CDCl3, ppm): d 7.44 (s, 1H), 7.26e7.04 (m, 3H), 4.20e4.07
(m, 2H), 2.21e2.15 (m, 2H), 2.07e1.92 (m, 3H), 1.84e1.35 (m, 8H),
1.31 (t, J ¼ 7.2 Hz, 3H), 1.22 (s, 3H), 1.20e1.16 (m, 2H), 1.06e0.99 (m,
1H), 0.97 (s, 3H), 0.95e0.87 (m, 2H), 0.60 (s, 3H); 13C NMR
4.3.8.23. Pyrazole derivative 11w. Yield 75%; Mp 157.4e158.6 ꢀC; IR
(KBr): 3096, 2942, 2852, 1626, 1593, 1534, 1495, 1465, 1417, 1377,
1319, 1228, 1160, 1129, 1025, 997, 855, 832, 808, 638 cmꢁ1; 1H NMR
(100 MHz, CDCl3, ppm):
d
177.4, 158.1 (dd, J ¼ 243.4, 2.6 Hz), 153.6,
152.3 (dd, J ¼ 245.1, 2.9 Hz),135.9,131.2,128.9 (dd, J ¼ 14.4,10.8 Hz),
117.0 (dd, J ¼ 22.5, 8.9 Hz), 115.9 (dd, J ¼ 22.4, 7.7 Hz), 115.2 (d,
J ¼ 25.7 Hz), 68.0, 60.0, 57.0, 52.6, 43.8, 42.7, 41.8, 40.2, 38.1, 38.1,
37.7, 34.6 (d, J ¼ 4.2 Hz), 29.0, 22.6, 21.3, 21.1, 19.0, 14.2, 12.7; HRMS
(ESI, m/z) calcd for C29H37F2N2O2 [M þ H]þ 483.2823. Found:
483.2821.
(400 MHz, CDCl3, ppm):
d
7.55 (d, J ¼ 2.4 Hz, 1H), 7.49 (d, J ¼ 8.6 Hz,
1H), 7.41 (s, 1H), 7.27 (dd, J ¼ 8.6, 2.4 Hz, 1H), 4.20e4.07 (m, 2H),
2.21e2.17 (m, 2H), 2.06e1.91 (m, 3H), 1.83e1.40 (m, 7H), 1.31 (t,
J ¼ 7.2 Hz, 3H), 1.25e1.24 (m, 1H), 1.22 (s, 3H), 1.21e1.16 (m, 2H), 1.10
(s, 3H), 1.06e0.88 (m, 3H), 0.57 (s, 3H); 13C NMR (100 MHz, CDCl3,
ppm):
d 177.4, 151.4, 139.2, 135.6, 132.8, 132.4, 130.8, 130.4, 125.1,
122.3, 68.0, 60.0, 57.0, 52.4, 43.8, 42.5, 41.7, 40.2, 38.1, 38.1, 37.6,
4.3.8.19. Pyrazole derivative 11s. Yield 92%; Mp 151.4e152.2 ꢀC; IR
(KBr): 3084, 2954, 2921, 2851, 1715, 1582, 1535, 1499, 1459, 1424,
1380, 1337, 1304, 1240, 1184, 1151, 1093, 1072, 1045, 998, 853, 794,
35.7, 29.0, 22.5, 22.0, 21.3, 19.0, 14.2, 12.6; HRMS (ESI, m/z) calcd for
C
29H37Cl2N2O2 [M þ H]þ 515.2232. Found: 515.2234; [Mþ2 þ H]þ
517.2232. Found: 517.2227.
777, 717, 524 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
; d 7.57e7.55
(m, 1H), 7.42 (s, 1H), 7.29e7.25 (m, 2H), 4.20e4.06 (m, 2H), 2.21e
2.17 (m, 2H), 2.08e1.97 (m, 3H),1.82e1.36 (m, 7H),1.31 (t, J ¼ 7.2 Hz,
3H), 1.27e1.25 (m, 1H), 1.23 (s, 3H), 1.21e1.15 (m, 2H), 1.06e0.90 (m,
3H), 0.85 (s, 3H), 0.63 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
4.3.8.24. Pyrazole derivative 11x. Yield 60%; Mp 204.9e206.6 ꢀC; IR
(KBr): 3076, 2957, 2927, 2851, 1711, 1593, 1530, 1490, 1450, 1381,
1362, 1237, 1182, 1152, 1111, 1096, 1026, 1000, 862, 806, 680 cmꢁ1
;
1H NMR (400 MHz, CDCl3, ppm):
d
7.41 (s, 1H), 7.34 (d, J ¼ 1.8 Hz,
d
177.4, 153.4, 139.4, 135.3, 133.8, 131.8, 131.0, 129.8, 127.8, 127.0,
2H), 7.30 (t, J ¼ 1.8 Hz, 1H), 4.22e4.05 (m, 2H), 2.21e2.17 (m, 2H),
67.2, 60.0, 57.1, 52.6, 43.8, 43.0, 41.2, 40.2, 38.1, 38.1, 37.8, 35.1, 29.7,
2.05e1.91 (m, 3H), 1.85e1.36 (m, 7H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.27e

S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
81
1.24 (m, 1H), 1.22 (s, 3H), 1.21e1.14 (m, 2H), 1.13 (s, 3H), 1.06e0.83
(m, 3H), 0.57 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
177.4,151.4,
141.4, 135.9, 135.1, 135.1, 132.7, 126.8, 121.6, 121.6, 68.1, 60.0, 57.0,
52.4, 43.8, 42.4, 41.7, 40.2, 38.1, 38.1, 37.6, 35.6, 29.0, 22.5, 22.1, 21.3,
19.0, 14.2, 12.6; HRMS (ESI, m/z) calcd for C29H37Cl2N2O2 [M þ H]þ
515.2232. Found: 515.2230; [Mþ2 þ H]þ 517.2232. Found: 517.2224.
J ¼ 1.6 Hz, 1H), 7.15 (d, J ¼ 8.0 Hz, 1H), 7.08 (dd, J ¼ 8.0, 2.0 Hz, 1H),
4.21e4.06 (m, 2H), 2.29 (s, 3H), 2.28 (s, 3H), 2.21e2.15 (m, 2H),
2.07e1.93 (m, 3H), 1.84e1.35 (m, 7H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.27e
1.25 (m, 1H), 1.22 (s, 3H), 1.20e1.16 (m, 2H), 1.10 (s, 3H), 1.02e0.87
d
(m, 3H), 0.60 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.4, 151.1,
137.7, 137.2, 135.6, 134.4, 131.0, 129.7, 124.8, 120.8, 67.9, 60.0, 57.1,
52.6, 43.8, 42.5, 41.4, 40.3, 38.2, 38.1, 37.8, 35.9, 29.0, 22.6, 21.9, 21.4,
19.8, 19.4, 19.0, 14.2, 12.7; HRMS (ESI, m/z) calcd for C31H43N2O2
[M þ H]þ 475.3325. Found: 475.3321.
4.3.8.25. Pyrazole derivative 11y. Yield 56%; Mp 139.1e140.6 ꢀC; IR
(KBr): 3087, 2955, 2870, 2850, 1717, 1583, 1529, 1506, 1469, 1448,
1386, 1234, 1179, 1149, 1092, 1042, 985, 852, 785, 718, 603 cmꢁ1; 1H
NMR (400 MHz, CDCl3, ppm):
d
7.36 (s, 1H), 7.20 (d, J ¼ 7.6 Hz, 1H),
4.3.8.30. Pyrazole derivative 11ad. Yield 81%; Mp 215.5e216.3 ꢀC;
7.11 (t, J ¼ 7.6 Hz, 1H), 7.04 (d, J ¼ 6.5 Hz, 1H), 4.20e4.08 (m, 2H),
2.31 (s, 3H), 2.21e2.15 (m, 2H), 2.09e1.96 (m, 3H),1.94 (s, 3H),1.82e
1.36 (m, 7H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.29e1.25 (m, 1H), 1.23 (s, 3H),
IR (KBr): 3088, 2955, 2850, 1712, 1605, 1526, 1496, 1463, 1404, 1379,
1323, 1234, 1182, 1152, 1091, 1030, 971, 852, 695 cmꢁ1 1H NMR
;
(400 MHz, CDCl3, ppm):
d 7.37 (s, 1H), 7.02 (s, 2H), 6.95 (s, 1H),
1.20e1.16 (m, 2H), 1.06e0.84 (m, 3H), 0.80 (s, 3H), 0.64 (s, 3H); 13
C
4.21e4.07 (m, 2H), 2.35 (s, 6H), 2.21e2.15 (m, 2H), 2.07e1.93 (m,
3H), 1.86e1.38 (m, 7H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.27e1.24 (m, 1H),
1.22 (s, 3H), 1.20e1.17 (m, 2H), 1.06 (s, 3H), 1.03e0.88 (m, 3H), 0.59
NMR (100 MHz, CDCl3, ppm):
d 177.5, 152.8, 139.0, 138.0, 134.7,
134.1, 130.2, 128.9,125.3,125.3, 67.2, 60.0, 57.2, 52.7, 43.8, 42.9, 40.9,
40.3, 38.2, 38.1, 38.0, 35.5, 29.0, 22.6, 21.7, 21.3, 20.2, 19.0, 14.2, 14.2,
12.7; HRMS (ESI, m/z) calcd for C31H43N2O2 [M þ H]þ 475.3325.
Found: 475.3327.
(s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.4, 151.1, 139.7, 138.5,
138.5, 134.4, 131.2, 128.7, 121.3, 121.3, 68.0, 60.0, 57.1, 52.6, 43.8,
42.5, 41.5, 40.3, 38.2, 38.1, 37.8, 35.9, 29.0, 22.6, 21.9, 21.4, 21.2, 21.2,
19.0, 14.1, 12.7; HRMS (ESI, m/z) calcd for C31H43N2O2 [M þ H]þ
475.3325. Found: 475.3322.
4.3.8.26. Pyrazole derivative 11z. Yield 63%; Mp 114.0e115.6 ꢀC; IR
(KBr): 3090, 2950, 2867, 2847,1719,1615,1519,1449,1377,1321,1302,
1235, 1180, 1151, 1094, 1041, 1029, 989, 847, 818, 653, 600 cmꢁ1; 1H
4.4. X-ray crystallographic analysis
NMR (400 MHz, CDCl3, ppm):
d 7.35 (s, 1H), 7.07 (s, 1H), 7.05 (d,
J ¼ 8.0 Hz,1H), 7.00 (d, J ¼ 8.0 Hz,1H), 4.20e4.07 (m, 2H), 2.35 (s, 3H),
2.21e2.14 (m, 2H), 2.06 (s, 3H), 2.04e1.96 (m, 3H),1.82e1.36 (m, 7H),
1.31 (t, J ¼ 7.2 Hz, 3H), 1.29e1.25 (m, 1H), 1.22 (s, 3H), 1.20e1.15 (m,
2H), 1.05e0.84 (m, 3H), 0.82 (s, 3H), 0.62 (s, 3H); 13C NMR (100 MHz,
X-ray crystal data of compounds 9b, 10, 11v and 11a were
collected by
monochromated Cu-K
a Rigaku AFC5R diffractometer with graphite-
ꢀ
a
radiation (
l
¼ 0.71073 A). The structure
was solved by the direct method and refined with a full-matrix
CDCl3, ppm):
d
177.5, 152.7, 138.6, 136.5, 135.6, 134.1, 131.3, 128.9,
least squares method.
127.2, 126.5, 67.2, 60.0, 57.2, 52.7, 43.8, 42.9, 40.9, 40.3, 38.2, 37.9,
35.5, 29.7, 29.0, 22.6, 21.6, 21.3, 21.1, 19.0, 17.1, 14.2, 12.7; HRMS (ESI,
m/z) calcd for C31H43N2O2 [M þ H]þ 475.3325. Found: 475.3322.
4.4.1. Crystal data for compound 9b
C
29H39FN2O2, M ¼ 466.62, orthorhombic, space group P21,
3
ꢀ
a ¼ 8.2822(9), b ¼ 7.9215(9), c ¼ 19.349(2), V ¼ 1268.4(2) A , Z ¼ 2,
4.3.8.27. Pyrazole derivative 11aa. Yield 79%; Mp 136.5e137.6 ꢀC;
IR (KBr): 3089, 2950, 2843, 1719, 1620, 1581, 1516, 1447, 1376, 1323,
1233, 1178, 1148, 1093, 1063, 1028, 974, 852, 827, 653, 606 cmꢁ1; 1H
m
(Cu K
a
) ¼ 0.644 cmꢁ1, F (000) ¼ 504, Dc ¼ 1.222 mg/mm3, crystal
dimensions: 0.30 ꢂ 0.28 ꢂ 0.25 mm, A total of 9627 reflections
were collected using the
u
ꢁ 2
q
scan technique to a maximum 2
q
NMR (400 MHz, CDCl3, ppm):
d
7.35 (s, 1H), 7.15 (d, J ¼ 8.0 Hz, 1H),
value, and 4364 reflections with I > 2s
(I) were used in the struc-
7.11 (d, J ¼ 8.0 Hz, 1H), 6.98 (s, 1H), 4.20e4.07 (m, 2H), 2.32 (s, 3H),
2.21e2.15 (m, 2H), 2.08e2.207 (m, 1H), 2.05 (s, 3H), 2.03e1.97 (m,
2H), 1.82e1.36 (m, 7H), 1.31 (t, J ¼ 7.2 Hz, 3H), 1.29e1.25 (m, 1H),
1.23 (s, 3H), 1.20e1.16 (m, 2H), 1.06e0.86 (m, 3H), 0.82 (s, 3H), 0.63
ture determination. Final R and Rw values were 0.0466 and 0.1146,
respectively. The maximum and minimum peaks in the difference
ꢀꢁ3
map were 0.122 and ꢁ0.115 e A , respectively. The data has been
deposited with the Cambridge Crystallographic Data Centre as
supplementary publication number CCDC 917721.
(s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
d 177.5, 152.5, 138.8, 135.7,
134.1, 132.6, 130.4, 129.5, 129.0, 128.0, 67.2, 60.0, 57.1, 52.7, 43.8,
42.9, 40.9, 40.3, 38.2, 38.1, 38.0, 35.5, 29.0, 22.6, 21.6, 21.3, 20.7, 19.0,
16.7, 14.2, 12.7; HRMS (ESI, m/z) calcd for C31H43N2O2 [M þ H]þ
475.3325. Found: 475.3323.
4.4.2. Crystal data for compound 10
C
23H34O4,
M
¼
374.50, orthorhombic, space group P21,
3
ꢀ
a ¼ 6.8882(14), b ¼ 11.523(2), c ¼ 13.459(3), V ¼ 1053.8(4) A , Z ¼ 2,
m
(Cu K
a
) ¼ 0.079 cmꢁ1, F (000) ¼ 408, Dc ¼ 1.180 mg/mm3, crystal
4.3.8.28. Pyrazole derivative 11ab. Yield 69%; Mp 173.1e174.0 ꢀC; IR
dimensions: 0.20 ꢂ 0.20 ꢂ 0.20 mm, A total of 3307 reflections
(KBr): 3088, 2950, 2847, 1602, 1529, 1505, 1467, 1449, 1377, 1299,
were collected using the
u
ꢁ 2
q
scan technique to a maximum 2
q
1235, 1180, 1150, 1095, 1026, 988, 852, 782, 578 cmꢁ1
;
1H NMR
value, and 1845 reflections with I > 2s
(I) were used in the structure
(400 MHz, CDCl3, ppm):
d
7.40 (s, 1H), 7.21 (t, J ¼ 7.6 Hz, 1H), 7.10e
determination. Final R and Rw values were 0.0663 and 0.1797,
7.07 (m, 2H), 4.22e4.07 (m, 2H), 2.21e2.13 (m, 2H), 2.10e2.04 (m,
2H), 2.02 (s, 3H), 2.01e1.98 (m, 1H), 1.97 (s, 3H), 1.85e1.35 (m, 8H),
1.31 (t, J ¼ 7.2 Hz, 3H), 1.23 (s, 3H), 1.21e1.17 (m, 2H), 1.06e0.85 (m,
3H), 0.78 (s, 3H), 0.65 (s, 3H); 13C NMR (100 MHz, CDCl3, ppm):
respectively. The maximum and minimum peaks in the difference
ꢀꢁ3
map were 0.283 and ꢁ0.250 e A , respectively. The data has been
deposited with the Cambridge Crystallographic Data Centre as
supplementary publication number CCDC 917818.
d
177.5, 152.3, 138.3, 136.9, 136.3, 134.4, 129.0, 128.7, 127.8, 127.7,
67.3, 60.0, 57.2, 52.5, 43.8, 42.9, 40.9,,40.3, 38.1, 38.1, 38.0, 35.2, 29.0,
4.4.3. Crystal data for compound 11v
22.6, 21.1, 20.7, 19.0, 17.1, 17.1, 14.2, 12.7; HRMS (ESI, m/z) calcd for
C
C
29H36Cl2N2O2, M ¼ 515.50, orthorhombic, space group P21,
3
ꢀ
31H43N2O2 [M þ H]þ 475.3325. Found: 475.3322.
a ¼ 12.6459(5), b ¼ 8.3831(3), c ¼ 13.3872(5), V ¼ 1340.69(9) A ,
Z ¼ 2,
m
(Cu K
a
) ¼ 2.396 cmꢁ1, F (000) ¼ 548, Dc ¼ 1.277 mg/mm3,
4.3.8.29. Pyrazole derivative 11ac. Yield 59%; Mp 124.1e125.3 ꢀC; IR
(KBr): 3091, 2930, 2869, 2847, 1719, 1613, 1586, 1512, 1449, 1377,
1338, 1321, 1239, 1181, 1151, 1092, 1029, 978, 879, 852, 817,
crystal dimensions: 0.24 ꢂ 0.20 ꢂ 0.20 mm, A total of 10,911 re-
flections were collected using the
maximum 2 value, and 4707 reflections with I > 2
in the structure determination. Final R and Rw values were 0.0644
u
ꢁ 2
q
scan technique to a
q
s
(I) were used
646 cmꢁ1 1H NMR (400 MHz, CDCl3, ppm):
; d 7.37 (s, 1H), 7.22 (d,

82
S.-L. Zhu et al. / European Journal of Medicinal Chemistry 65 (2013) 70e82
[15] F.R. Chang, P.Y. Yang, J.Y. Lin, K.H. Lee, Y.C. Wu, Bioactive kaurane diterpenoids
from annona glabra, J. Nat. Prod. 61 (1998) 437e439.
[16] K.L. Wong, P. Chan, H.Y. Yang, F.L. Hsu, I.M. Liu, Y.W. Cheng, J.T. cheng, Iso-
steviol acts on potassium channels to relax isolated aortic strips of Wistar rat,
Life Sci 74 (2004) 2379e2387.
[17] W.L. Braguinu, M.A.B. Gomes, B.H. Oliveira, E.G.S. Garnieri, M.E.M. Rocha,
M.B.M. Oliveira, Activity of isosteviol lactone on mitochondrial metabolism,
Toxicol. Lett. 143 (2003) 83e92.
and 0.1809, respectively. The maximum and minimum peaks in the
difference map were 0.156 and ꢁ0.236 e Åꢁ3, respectively. The data
has been deposited with the Cambridge Crystallographic Data
Centre as supplementary publication number CCDC 917722.
4.4.4. Crystal data for compound 11a
[18] O. Wonganan, C. Tocharus, C. Puedsing, S. Homvisasevongsa, O. Sukcharoen,
A. Suksamrarn, Potent vasorelaxant analogs from chemical modification and
biotransformation of isosteviol, Eur. J. Med. Chem. 62 (2013) 771e776.
[19] V.E. Kataev, O.I. Militsina, I.Y. Strobykina, G.I. Kovylyaeva, R.Z. Musin,
O.V. Fedorova, G.L. Rusinov, M.N. Zueva, G.G. Mordovskoi, A.G. Tolstikov,
Synthesis and anti-tuberculous activity of diesters based on isosteviol and
dicarboxylic acids, Pharm. Chem. J. 40 (2006) 473e475.
[20] M.G. Korochkina, A.D. Nikitashina, R.N. Khaybullin, K.A. Petrov,
I.Yu. Strobykina, V.V. Zobov, V.E. Kataev, Unfolded and macrocyclic ammo-
nium derivatives of diterpenoids steviol and isosteviol having choline moi-
eties. Synthesis and inhibitory activities toward acetylcholine- and
butyrylcholinesterases, Med. Chem. Commun. 3 (2012) 1449e1454.
[21] J. Li, D.Y. Zhang, X.M. Wu, Synthesis and biological evaluation of novel exo-
methylene cyclopentanone tetracyclic diterpenoids as antitumor agents,
Bioorg. Med. Chem. Lett. 21 (2011) 130e132.
C
29H38N2O2, M ¼ 446.61, orthorhombic, space group P212121,
3
ꢀ
a ¼ 7.0203(14), b ¼ 16.816(3), c ¼ 21.249(4), V ¼ 2508.5(9) A , Z ¼ 4,
m
(Cu K
a
) ¼ 0.074 cmꢁ1, F (000) ¼ 968, Dc ¼ 1.183 mg/mm3, crystal
dimensions: 0.20 ꢂ 0.18 ꢂ 0.18 mm, A total of 7545 reflections were
collected using the
u
ꢁ 2q
scan technique to a maximum 2
q value,
and 4146 reflections with I > 2
s
(I) were used in the structure
determination. Final R and Rw values were 0.0647 and 0.1320,
respectively. The maximum and minimum peaks in the difference
map were 0.163 and ꢁ0.154 e Åꢁ3, respectively. The data has been
deposited with the Cambridge Crystallographic Data Centre as
supplementary publication number CCDC 924756.
[22] Y.F. Zeng, J.Q. Wu, L.Y. Shi, K. Wang, B. Zhou, Y. Tang, D.Y. Zhang, Y.C. Wu,
Acknowledgments
W.Y. Hua, X.M. Wu, Synthesis and evaluation of cytotoxic effects of novel a-
methylenelactone tetracyclic diterpenoids, Bioorg. Med. Chem. Lett. 22 (2012)
1922e1925.
[23] Y. Wu, G.F. Dai, J.H. Yang, Y.X. Zhang, Y. Zhu, J.C. Tao, Stereoselective synthesis
of 15- and 16-substituted isosteviol derivatives and their cytotoxic activities,
Bioorg. Med. Chem. Lett. 19 (2009) 1818e1821.
[24] T. Zhang, L.H. Lu, H. Liu, J.W. Wang, R.X. Wang, Y.X. Zhang, J.C. Tao, D-ring
modified novel isosteviol derivatives: design, synthesis and cytotoxic activity
evaluation, Bioorg. Med. Chem. Lett. 22 (2012) 5827e5832.
The authors are grateful for the financial support to the National
Natural Science Foundation of China (Project No. 20772113) and the
Doctoral Research fund of Henan University of Traditional Chinese
Medicine for financial support (No. BSJJ2009-41).
[25] M.F. Brana, J.M. Castellano, M. Mpran, M.J. Perez de Vega, X.D. Gian,
C.A. Romerdahl, G. Keihauer, Bis-naphthalimides. 2. synthesis and biological
activity of 5,6-acenaphthalimidoalkyl-1,8-naphthalimidoalkyl amines, Eur. J.
Med. Chem. 30 (1995) 235e239.
Appendix A. Supplementary data
Supplementary data related to this article can be found at
http://dx.doi.org/10.1016/j.ejmech.2013.04.044.
[26] N. Kojima, T. Fushimi, N. Maezaki, T. Tanaka, T. Yamori, Synthesis of hybrid
acetogenins, a,b-unsaturated-g-lactone-free nitrogen-containing heterocyclic
analogues, and their cytotoxicity against human cancer cell lines, Bioorg. Med.
Chem. Lett. 18 (2008) 1637e1641.
References
[27] A.E.G.E. Amr, N.A. Abdel-Latif, M.M. Abdalla, Synthesis and antiandrogenic
activity of some new 3-substituted androstano[17,16-c]-50-aryl-pyrazoline
and their derivatives, Bioorg. Med. Chem. 14 (2006) 373e384.
[28] J.H. Ahn, H.M. Kim, S.H. Jung, S.K. Kang, K.R. Kim, S.D. Rhee, S.D. Yang,
H.G. Cheon, S.S. Kim, Synthesis and DP-IV inhibition of cyano-pyrazoline de-
rivatives as potent anti-diabetic agents, Bioorg. Med. Chem. Lett. 14 (2004)
4461e4465.
[29] P. Cui, T.L. Macdonald, M. Chen, J.L. Nadler, Synthesis and biological evaluation
of lisofylline (LSF) analogs as a potential treatment for Type 1 diabetes, Bioorg.
Med. Chem. Lett. 16 (2006) 3401e3405.
[30] A.R. Renslo, G.W. Luehr, S. Lam, N.E. Westlund, M. Gómez, C.J. Hackbarth,
D.V. Patel, M.F. Gordeev, Synthesis and structureeactivity studies of anti-
bacterial oxazolidinones containing dihydrothiopyran or dihydrothiazine C-
rings, Bioorg. Med. Chem. Lett. 16 (2006) 3475e3478.
[31] E. Aiello, S. Aiello, F. Mingoia, A. Bacchi, C. Pelizzi, C. Musiu, S.M. Giovanna,
A. Pani, C.P. La, M.M. Elena, Synthesis and antimicrobial activity of new 3-(1-R-
3(5)-methyl-4-nitroso-1H-5(3)-pyrazolyl)-5-methylisoxazoles, Bioorg. Med.
Chem. 8 (2000) 2719e2728.
[1] M.A. Metwally, M.A. Gouda, A.N. Harmal, A.M. Khalil, Synthesis, antitumor,
cytotoxic and antioxidant evaluation of some new pyrazolotriazines attached
to antipyrine moiety, Eur. J. Med. Chem. 56 (2012) 254e262.
[2] S. Sangthong, K. Krusong, N. Ngamrojanavanich, T. Vilaivan, S. Puthong,
S. Chandchawan, N. Muangsin, Synthesis of rotenoid derivatives with cyto-
toxic and topoisomerase II inhibitory activities, Bioorg. Med. Chem. Lett. 21
(2011) 4813e4818.
[3] R.H. Cao, W.X. Fan, L. Guo, Q. Ma, G.X. Zhang, J.R. Li, X.M. Chen, Z.H. Ren,
L.Q. Qiu, Synthesis and structureeactivity relationships of harmine derivatives
as potential antitumor agents, Eur. J. Med. Chem. 60 (2013) 135e143.
[4] M.G. Korochkina, V.M. Babaev, I.Y. Strobykina, A.D. Voloshina, N.V. Kulik,
V.E. Kataev, Synthesis and antimicrobial activity of several bis-quaternized
ammonium derivatives of the diterpenoid isosteviol, Chem. Nat. Compd. 47
(2012) 914e917.
[5] D. Biedermann, B. Eigenrova, M. Hajduch, J. Sarek, Synthesis and evaluation of
biological activity of the quaternary ammonium salts of lupane-, oleanane-,
and ursane-type acids, Synthesis 22 (2010) 3839e3848.
[6] L.F. Tietze, H.P. Bell, S. Chandrasekhar, Natural product hybrids as new leads
for drug discovery, Angew. Chem. Int. Ed. 42 (2003) 3996e4028.
[7] Y.Y. Guan, D. Zheng, Z. Yan, N. Wang, P.S. Lei, Synthesis and antitumor activity
of 5,6-dihydro-17-hydroxy icogenin analogs, Eur. J. Med. Chem. 51 (2012)
200e205.
[8] A.D. Kinghorn, D.D. Soejarto, N.P.D. Nanayakkara, C.M. Compadre, H.C. Makapugay,
J.M. Hovanec-Brown, P.J. Medon, S.K. Kamath, A phytochemical screening proce-
dure for sweet ent-kaurene glycosides in the genus stevia, J. Nat. Prod. 47 (1984)
439e444.
[32] N.R. Mohamed, G.A. Elmegeed, H.A. Abd-ElMalek, M. Younis, Synthesis of
biologically active steroid derivatives by the utility of Lawesson’s reagent,
Steroids 70 (2005) 131e136.
[33] I. Chaaban, E. El-Khawass, M. Mahran, O. El-Sayed, H. El-Saidi, H. Aboul-Enen,
Design, synthesis, and in vitro evaluation of cytotoxic activity of new
substituted 1,4-benzoquinones and hydroquinones, Med. Chem. Res. 16
(2007) 49e77.
[34] P.M. Dewang, D.K. Kim, Synthesis and biological evaluation of 2-pyridyl-
substituted pyrazoles and imidazoles as transforming growth factor-b type 1
[9] M.S. Melis, Renal excretion of stevioside in rats, J. Nat. Prod. 55 (1992) 688e690.
[10] K. Yasukawa, S. Kitanaka, S. Seo, Inhibitory effect of stevioside on tumor
promotion by 12-o-tetradecanoylphorbol-13-acetate in two-stage carcino-
genesis in mouse skin, Biol. Pharm. Bull. 25 (2002) 1488e1490.
[11] S.F. Chang, L.M. Yang, C.H. Lo, J.H. Liaw, L.H. Wang, S.J. Lin, Microbial trans-
formation of isosteviol and bioactivities against the glucocorticoid/androgen
response elements, J. Nat. Prod. 71 (2008) 87e92.
[12] J.C. Liu, P.F. Kao, M.H. Hsieh, Y.J. Chen, P. Chan, Isolation and identification of
endophytic from eucommia ulmoides and their biotransformation activity of
isosteviol, Acta Cardiol. Sin. 17 (2001) 133e140.
[13] D.Y. Xu, S.J. Zhang, J.R. David, J.P. Wang, The effects of isosteviol against
myocardium injury induced by ischaemia-reperfusion in the isolated guinea
pig heart, Clin. Exp. Pharmacol. Physiol. 34 (2007) 488e493.
[14] K.L. Wong, J.W. Lin, J.C. Liu, H.Y. Yang, P.F. Kao, C.H. Chen, S.H. Loh, W.T. Chiu,
T.H. Cheng, J.G. Lin, H.J. Hong, Antiproliferative effect of isosteviol on
angiotensin-II-treated rat aortic smooth muscle cells, Pharmacology 76 (2006)
163e169.
receptor kinase inhibitors, Bioorg. Med. Chem. Lett. 20 (2010) 4228e4232.
[35] C.H. Jin, M. Krishnaiah, D. Sreenu, V.B. Subrahmanyam, K.S. Rao, C.Y. Park,
J.Y. Son, Y.Y. Sheen, D.K. Kim, Synthesis and biological evaluation of 1-
substituted-3-(6-methylpyridin- 2-yl)-4-([1,2,4]triazolo[1,5-a]pyridin-6-yl)
pyrazoles as transforming growth factor-b type 1 receptor kinase inhibitors,
Bioorg. Med. Chem. Lett. 21 (2011) 6049e6053.
[36] E. Frank, Z. Mucsi, I. Zupko, B. Rethy, G.F.G. Schneider, J. Wolfling, Efficient
approach to androstene-fused arylpyrazolines as potent antiproliferative
agents. Experimental and theoretical studies of substituent effects on BF3-
catalyzed intramolecular [3þ2] cycloadditions of olefinic phenylhydrazones,
J. Am. Chem. Soc. 131 (2009) 3894e3904.
[37] E. Frank, J. Wolfling, B. Aukszi, V. Konig, T.R. Schneider, G. Schneider, Ster-
eoselective synthesis of some novel heterocyclic estrone derivatives by
intramolecular 1,3-dipolar cycloaddition, Tetrahedron 58 (2002) 6843e6849.
[38] J.C. Tao, G.Q. Tian, Y.B. Zhang, Y.Q. Fu, G.F. Dai, Y. Wu, Synthesis and bioactivity of
isosteviol derivatives: a facile method for preparation of ent-16a-hydroxy-15b-
hydroxymethylbeyeran-19-oic Acid, Chin. Chem. Lett. 16 (2005) 1441e1444.