Design, synthesis and biological evaluation of some novel N-arylpyrazole derivatives bearing the sulfonamide moiety as cytotoxic agents

Article abstract of DOI:10.1007/s11164-016-2620-x

A series of novel N-arylpyrazole derivatives (4a–4l) bearing the sulfonamide moiety were synthesized by the condensation reaction of 1,3-dicarbonyl compounds with 4-hydrazinylbenzenesulfonamide. The structures of the obtained compounds were established on

Full text of DOI:10.1007/s11164-016-2620-x

Res Chem Intermed
DOI 10.1007/s11164-016-2620-x
Design, synthesis and biological evaluation of some
novel N-arylpyrazole derivatives bearing
the sulfonamide moiety as cytotoxic agents
1
,2
1,2
1,2
1,2
•
Xiaobo Duan
1
•
Yingxing Wang
•
Weipei Feng
1,2
Xiaobing Yang
•
Yaxing Yang
Jinchao Zhang
,2
1,2
1,2
•
Hongyan Li
Shuxiang Wang
•
Shenghui Li
,2
•
•
1
1,2
1,2
•
Guoqiang Zhou
•
Chuanqi Zhou
Received: 25 January 2016 / Accepted: 22 June 2016
Ó Springer Science+Business Media Dordrecht 2016
Abstract A series of novel N-arylpyrazole derivatives (4a–4l) bearing the sulfon-
amide moiety were synthesized by the condensation reaction of 1,3-dicarbonyl
compounds with 4-hydrazinylbenzenesulfonamide. The structures of the obtained
compounds were established on the basis of elemental (C, H, and N) and spectral
1
13
analysis ( H NMR, C NMR, ESIMS, and FT-IR). These compounds were tested
for their in vitro cytotoxic activity against three human tumor cell lines: MCF-7,
Hela, and A549. The results showed that most of the obtained compounds exhibited
promising cytotoxicity against the tested cell lines with low IC50 values. The
pyrazole derivative 4k, bearing two methoxy groups on the 3-position and 4-posi-
tion of the phenyl ring, was the most effective one. Its inhibition of cell growth of
MCF-7 cells was better than that of celecoxib and cisplatin.
Keywords N-arylpyrazole Á Sulfonamide Á Anticancer Á Cytotoxicity Á Cancer cell
lines
Electronic supplementary material The online version of this article (doi:10.1007/s11164-016-2620-x)
contains supplementary material, which is available to authorized users.
&
&
&
1
Shenghui Li
lish@hbu.edu.cn
Jinchao Zhang
jczhang6970@yahoo.com.cn
Shuxiang Wang
wsx@hbe.edu.cn
Key Laboratory of Chemical Biology of Hebei Province, College of Chemistry and
Environmental Science, Hebei University, Baoding 071002, China
2
Key Laboratory of Medicinal Chemistry and Molecular Diagnosis, Ministry of Education,
Hebei University, Baoding 071002, China
123

X. Duan et al.
Introduction
Cancer is one of the most dreadful diseases in developed countries and many other
parts of the world, currently remains the second leading cause of death and is
expected to surpass heart disease as the leading cause in the next few years [1]. This
serious major public health problem brings an enormous burden on society all over
the world. Therefore, the discovery of selective, efficient, and safe drugs for cancer
chemotherapy remains an urgent and high priority for medicinal research.
The presence of the pyrazole motif in commercial drugs or drug candidates has
made this heterocycle a popular synthetic target for pharmaceutical and agrochem-
ical sciences [2, 3]. Among the various pyrazoles, N-arylpyrazole derivatives play
an important role in medicinal chemistry. Indeed, anti-inflammatory [4–7],
antinociceptive [8], anti-obesity [9, 10], anti-allodynic [11], anticancer [12], and
anti-HIV [13, 14] agents bearing the N-arylpyrazole moiety have been widely used
as therapeutical drugs toward major therapeutical targets, such as cyclooxygenase
(
hCB receptor [9, 10], COX-2/sEH [11], estrogen receptor [12] or HIV-reverse
COX) [4–6], p38a MAP-kinase [7], r receptor (r R) [8], cannabinoid hCB and
1
1
1
2
transcriptase [13, 14], respectively.
Sulfonamides constitute an important class of compounds which over the years
have had different applications as pharmacological agents in the treatment of a wide
variety of diseases [15, 16]. Indeed, they have also been reported to possess various
types of biological properties such as anticancer [17], antibacterial [18], antimi-
crobial [19, 20], antimalarial [21], antihypertensive [22], antitumor [23], and anti-
inflammatory activities [24–31], among others. Among these, the anticancer activity
was found to take place through a variety of mechanisms such as disruption of the
microtubule assembly, cell cycle arrest in the G1 phase, functional suppression of
the transcriptional activator NF-Y, angiogenesis and carbonic anhydrase inhibition
[15, 16]. The most prominent mechanism was the inhibition of carbonic anhydrase
isozymes [32–35].
Moreover, a host of structurally novel N-arylpyrazole derivatives bearing the
sulfonamide moiety have attracted considerable attention as they showed promising
anticancer and antitumor properties, both in vitro and in vivo [36]. In fact, the most
commonly used anti-inflammatory drug, celecoxib (4-[5-(4-methylphenyl)-3-(tri-
fluoromethyl)-1H-pyrazol-1-yl]benzenesulfamide), possessing these structural fea-
tures, displays important inhibitory capacity on not only cyclooxygenase-2 (COX-2)
but also cancer-related human carbonic anhydrase IX (hCA IX) [37]. All these
findings encouraged us to explore the synthesis of N-arylpyrazole derivatives
bearing the sulfonamide moiety as potential cytotoxic agents. In this paper, novel N-
arylpyrazole derivatives bearing the sulfonamide moiety were designed and
synthesized by condensation reaction of 1,3-dicarbonyl compounds with 4-hy-
drazinylbenzenesulfonamide (Scheme 1). The cytotoxic activity of the obtained
compounds 4a–4l was evaluated in vitro against MCF-7, Hela, and A549 cells by
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and
found to possess moderate activity.
1
23

Design, synthesis and biological evaluation of some novel…
Scheme 1 Reagents and condition: (i) H
2
O, HCl, 80 °C; (ii) NaNO
; (vi) NaClO ; (vii) NaOH; (viii) 4-hydrazinylbenzenesulfonamide
= R = R = H; 4b: R = CH , R = R = H; 4c: R = CF
= H; 4e: R = R = H; 4f: R = Cl, R = R = H;
= OH, R = H; 4i: R , R = R = H; 4j:
= R = OCH = OCH
2
3
, below 0 °C; (iii) NaHSO , below
0
°C (iv) HCl, reflux; (v) DMF, POCl
hydrochloride, ethanol, reflux. 4a: R
= R = H; 4d: R = NO , R
g: R = Br, R = R = H; 4h: R
= OCH , R = R
3
4
1
2
3
2
3
1
3
2
3
,
R
4
R
1
3
2
2
1
= R
3
2
= F, R
1
3
2
1
3
2
1
3
2
1
= R
3
2
= OCH
= R
3
1
3
1
3
2
3
= H; 4k: R
1
2
3
, R
3
= H; 4l: R
1
= R
2
3
3
Experimental
Materials and methods
RPMI-1640 medium, trypsin and fetal bovine serum were purchased from Gibco.
MTT, benzylpenicillin and streptomycin were from Sigma. Three different human
carcinoma cell lines, MCF-7, Hela and A549, were obtained from the American
Type Culture Collection. Mps were measured on a XT-4 microscopic melting-point
1
13
spectrometer and are uncorrected. H NMR and C NMR spectra were recorded on
a Bruker AVIII 600 NMR spectrometer. IR spectra were recorded using KBr pellets
and a Perkin-Elmer Model-683 spectrophotometer. The mass spectra were measured
by LC–MS apparatus (Agilent 1200-6310). Elemental analysis was determined on
an Elementar Vario EL III elemental analyzer. 4-Hydrazinylbenzenesulfonamide
hydrochloride was synthesized from 4-aminobenzenesulfonamide by diazotization
and then reduction with sodium bisulfite in an overall yield of 44 % [38].
General procedure for the preparation of dialdehyde compounds 3a–3l [39–41]
POCl (15.0 mmol) was added dropwise to dry DMF (6 mL) at 5–10 °C with
3
constant stirring. The mixture was stirred at room temperature for an additional
hour. Then, substituted phenylacetic acid (4.95 mmol) 1a–1l was added at once and
the resulting clear solution was stirred for 4 h at 95 °C and then at room temperature
overnight. The excess vilsmeyer reagent was decomposed with crushed ice, and
123

X. Duan et al.
.
then a saturated solution of NaClO H O 2.21 g (15 mmol) was added with stirring.
2
4
The resulting nearly white crystalline deposit of the perchlorate salt 2a–2l was
filtered and washed with two 2.3-mL portions of water. This compound was used in
the next step without further purification.
The perchlorate salt 2a–2l (7.5 mmol) was added to a warm solution of 600 mg
15 mmol) NaOH in 5 mL water, and the mixture was heated with stirring for
(
1
5 min (bath temperature 90 °C) until total dissolution of the organic salt was
observed. Then, the reaction mixture was gradually cooled to room temperature and
diluted with ice-water, acidified by dilute hydrochloric acid, and extracted with
CH Cl . The organic phase was dried over anhydrous Na SO , filtrated, and then
2
2
2
4
concentrated to give 1,3-dialdehydes compound as an oil-like crude product.
General procedure for compounds 4a–4l
4
-Hydrazinylbenzenesulfonamide hydrochloride (201 mg, 0.9 mmol) was added to
the stirred solution of aryl malondialdehyde (1 mmol) in 15 mL ethanol. The
mixture was heated to reflux for 4 h. Then, the reaction mixture was cooled to room
temperature and the precipitate was filtrated, dried, and recrystallized using ethyl
alcohol as solvent.
4
-(4-Phenyl-1H-pyrazol-1-yl)benzenesulfonamide 4a Pink solid; yield 71.7 %;
1
-
m.p. 258.1–259.2 °C; IR (KBr, cm ): 3326, 3126, 3068, 1598, 1510, 1404, 1336,
1
155, 1099, 1037, 958, 906, 867, 829, 757, 691, 615, 544, 503; H NMR (600 MHz,
1
DMSO-d ) d: 9.14 (s, 1H, pyrazole C -H), 8.33 (s, 1H, pyrazole C -H), 8.10 (d,
6
3
5
J = 9.0 Hz, 2H, Ar), 7.97 (d, J = 9.0 Hz, 2H, Ar), 7.75 (dd, J = 7.8 Hz,
J = 1.2 Hz, 2H, Ar), 7.45 (s, 2H, SO NH ), 7.44 (t, J = 7.8 Hz, 2H, Ar), 7.29 (tt,
1
2
2
2
1
3
J = 7.4 Hz, J = 1.2 Hz, 1H, Ar); C NMR (150 MHz, DMSO-d ) d: 141.56,
1
2
6
1
MS(ESI): 300.9 [M ? H] 322.3 [M ? Na] . Anal. calcd for C H N O S: C
41.25, 139.60, 131.25, 128.96, 127.36, 126.94, 125.38, 124.85, 124.82, 118.14;
?
?
1
5 13 3 2
6
0.19 H 4.38 N 14.04 Found: C 60.18 H 4.25 N 13.69. H O.
2
4
8
1
-(4-(p-Tolyl)-1H-pyrazol-1-yl)benzenesulfonamide 4b Light yellow solid; yield
-
1
1.4 %; m.p. 263.8–264.3 °C; IR (KBr, cm ): 3350, 3255, 1598, 1504, 1429,
402, 1307, 1199, 1162, 1099, 954, 908, 876, 822, 752, 524, 449; H NMR
1
(
600 MHz, DMSO-d ) d: 9.08 (s, 1H, pyrazole C -H), 8.28 (s, 1H, pyrazole C -H),
6
3
5
8
.08 (d, J = 9.0 Hz, 2H, Ar), 7.96 (d, J = 9.0 Hz, 2H, Ar), 7.64 (d, J = 8.4 Hz,
1
3
2
H, Ar), 7.44 (s, 2H, SO NH ), 7.24 (d, J = 8.4 Hz, 2H, Ar), 2.33 (s, 3H, CH );
2
C
2
3
NMR (150 MHz, DMSO-d ) d: 141.58, 141.15, 139.50, 136.19, 129.50, 128.37,
6
?
27.34, 125.30, 124.85, 124.42, 118.06, 20.68; MS(ESI): 336.7 [M ? Na] . Anal.
1
calcd for C H N O S: C 61.32 H 4.82 N13.41, Found: C 61.73 H 4.28 N 13.33.
1
6 15 3 2
4
-(4-(4-(Trifluoromethyl)phenyl)-1H-pyrazol-1-yl)benzenesulfonamide 4c Yellow
-
1
solid; yield 62.0 %; m.p. 267.2–268.0 °C; IR (KBr, cm ): 3340, 3134, 3076, 1596,
517, 1405, 1336, 1157, 1122, 1064, 1016, 960, 902, 841, 760, 694, 622, 543, 493;
1
1
H NMR (600 MHz, DMSO-d ) d: 9.31 (s, 1H, pyrazole C -H), 8.45 (s, 1H,
6
3
1
23

Design, synthesis and biological evaluation of some novel…
pyrazole C -H), 8.11 (d, J = 8.4 Hz, 2H, Ar), 7.98 (d, J = 8.4 Hz, 2H, Ar), 7.97 (d,
5
1
J = 8.4 Hz, 2H, Ar), 7.80 (d, J = 8.4 Hz, 2H, Ar), 7.45 (s, 2H, SO NH ); C NMR
3
2
2
(
150 MHz, DMSO-d ) d: 141.55, 141.38, 139.86, 135.53, 127.37, 127.01 (q,
6
2
1
JCF = 33 Hz), 125.97, 125.83, 125.79, 124.28 (q, J = 270 Hz), 123.38, 118.34;
CF
?
MS(ESI): 390.3 [M ? Na] . Anal. Calcd for C H F N O S: C 52.31 H 3.29
N11.44, Found: C 52.28 H 3.04 N 11.26.
1
6 12 3 3 2
4
7
1
-(4-(4-Nitrophenyl)-1H-pyrazol-1-yl)benzenesulfonamide 4d Gold solid; yield
-
1
0.0 %; m.p. 262.0–263.0 °C; IR (KBr, cm ): 3315, 3154, 1594, 1508, 1402,
338, 1271, 1166, 1108, 903, 856, 833, 754, 620, 551, 441; H NMR (600 MHz,
1
DMSO-d6) d: 9.44 (s, 1H, pyrazole C -H), 8.52 (s, 1H, pyrazole C -H), 8.30 (d,
3
5
J = 8.4 Hz, 2H, Ar), 8.12 (d, J = 9.0 Hz, 2H, Ar), 8.04 (d, J = 9.0 Hz, 2H, Ar),
1
3
7
.99 (d, J = 8.4 Hz, 2H, Ar), 7.48 (s, 2H, SO NH ); C NMR (150 MHz, DMSO-
2 2
d ) d: 145.70, 141.74, 141.24, 140.18, 138.40, 127.39, 126.78, 126.04, 124.32,
6
?
.
122.85, 118.46; MS(ESI): 367.9 [M ? Na] . Anal. Calcd for C H N O S 0.25-
15 12 4 4
CH CH OH: C 52.32 H 3.51 N16.27, Found: C 52.35 H 3.80 N 15.76.
3
2
4
-(4-(4-Fluorophenyl)-1H-pyrazol-1-yl)benzenesulfonamide 4e Light yellow
-
1
solid; yield 87.9 %; m.p. 251.0–251.7 °C; IR (KBr, cm ): 3326, 3131, 1596,
1
1
502, 1402, 1337, 1230, 1159, 957, 831, 770, 598, 542, 517; H NMR (600 MHz,
DMSO-d ) d: 9.13 (s, 1H, pyrazole C -H), 8.31 (s, 1H, pyrazole C -H), 8.08 (d,
6
3
5
J = 9.0 Hz, 2H, Ar), 7.97 (d, J = 9.0 Hz, 2H, Ar), 7.78 (dd, J = 9.0 Hz,
1
1
3
J = 5.4 Hz, 2H, Ar), 7.45 (s, 2H, SO NH ), 7.28 (t, J = 9.0 Hz, 2H, Ar);
2
C
2
2
1
NMR (150 MHz, DMSO-d ) d: 161.16 (d, JCF = 243.0 Hz), 141.52, 141.28,
6
4
3
1
39.54, 127.84 (d, J = 3.0 Hz), 127.36, 127.31 (d, J = 9.0 Hz), 124.80,
CF CF
2
23.90, 118.14, 115.76 (d, J = 22.5 Hz); MS(ESI): 340.7 [M ? Na] . Anal.
?
1
Calcd for C H FN O S: C 56.77 H 3.81 N13.24, Found: C 57.11 H 3.52 N 13.12.
CF
1
5
12
3 2
4
-(4-(4-Chlorophenyl)-1H-pyrazol-1-yl)benzenesulfonamide 4f Light yellow
1
-
solid; yield 67.6 %; m.p. 254.0–255.5 °C; IR (KBr, cm ): 3384, 3336, 3268,
120, 1593, 1487, 1402, 1323, 1155, 1097, 951, 831, 756, 662, 627, 540, 513, 442;
3
1
H NMR (600 MHz, DMSO-d ) d: 9.18 (s, 1H, pyrazole C -H), 8.35 (s, 1H,
6
3
pyrazole C -H), 8.08 (d, J = 8.4 Hz, 2H, Ar), 7.97 (d, J = 8.4 Hz, 2H, Ar), 7.78 (d,
5
1
J = 7.8 Hz, 2H, Ar), 7.50 (d, J = 7.8 Hz, 2H, Ar), 7.45 (s, 2H, SO NH ); C NMR
3
2
2
(
150 MHz, DMSO-d ) d: 141.46, 141.37, 139.62, 131.25, 130.26, 128.89, 127.36,
6
?
27.08, 125.16, 123.65, 118.20; MS(ESI): 357.4 [M ? Na] . Anal. Calcd for
1
C H ClN O S: C 53.98 H 3.62 N12.59, Found: C 54.16 H 3.29 N 12.45.
1
5
12
3 2
4
-(4-(4-Bromophenyl)-1H-pyrazol-1-yl)benzenesulfonamide 4g Brownish yellow
-
1
solid; yield 59.0 %; m.p. 276.4–276.8 °C; IR (KBr, cm ): 3338, 3247, 1596, 1510,
1
1
403, 1336, 1267, 1159, 1103, 1068, 954, 823, 660, 539, 505, 416; H NMR
(
600 MHz, DMSO-d ) d: 9.19 (s, 1H, pyrazole C -H), 8.35 (s, 1H, pyrazole C -H),
6
3
5
8
.08 (d, J = 9.0 Hz, 2H, Ar), 7.97 (d, J = 9.0 Hz, 2H, Ar), 7.72 (d, J = 8.4 Hz,
1
H, Ar), 7.63 (d, J = 8.4 Hz, 2H, Ar), 7.44 (s, 2H, SO NH ); C NMR (150 MHz,
3
2
2
2
DMSO-d ) d: 141.45, 141.38, 139.59, 131.79, 130.63, 127.40, 127.36, 125.17,
6
123

X. Duan et al.
?
23.68, 119.68, 118.21; ESI–MS m/z: 400.1 [M ? Na] . Anal. Calcd for
1
C H BrN O S: C 47.63 H 3.20 N11.11, Found: C 48.00 H 2.93 N 10.99.
1
5
12
3 2
4
-(4-(4-Hydroxyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide 4h Brownish yel-
-
1
low solid; yield 58.1 %; m.p. 301.8–304 °C; IR (KBr, cm ): 3321, 3127, 1597,
1
6
8
504, 1402, 1344, 1298, 1267, 1199, 1149, 1097, 1027, 952, 910, 831, 787, 750,
1
61, 607, 548, 445; H NMR (600 MHz, DMSO-d ) d: 9.57 (s, 1H, pyrazole C -H),
6
3
.97 (s, 1H,pyrazole C -H), 8.18 (s, 1H, OH), 8.07 (d, J = 9.0 Hz, 2H, Ar), 7.95 (d,
5
J = 9.0 Hz, 2H, Ar), 7.54 (d, J = 8.4 Hz, 2H, Ar), 7.44 (s, 2H, SO NH ), 6.83 (d,
2
2
1
3
J = 8.4 Hz, 2H, Ar); C NMR (150 MHz, DMSO-d ) d: 156.43, 141.64, 140.96,
6
1
39.27, 127.34, 126.71, 125.08, 123.62, 122.15, 117.91, 115.72; MS(ESI): 338.6
?
M ? Na] .
[
Anal. Calcd for C H N O S: C 57.13 H 4.16 N13.33, Found: C 56.99 H 4.26
1
5 13 3 3
N13.23.
4
-(4-(4-Methoxyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide 4i Yellowish green
-
1
solid; yield 75.3 %; m.p. 237.8–241.3 °C; IR (KBr, cm ): 3370, 3264, 1601, 1571,
402, 1338, 1261, 1157, 1097, 1029, 952, 914, 825, 790, 752, 661, 605, 543, 457;
1
1
H NMR (600 MHz, DMSO-d ) d: 9.03 (s, 1H, pyrazole C -H), 8.25 (s, 1H,
6
3
pyrazole C -H), 8.08 (d, J = 9.0 Hz, 2H, Ar), 7.96 (d, J = 9.0 Hz, 2H, Ar), 7.67 (d,
J = 8.4 Hz, 2H, Ar), 7.44 (s, 2H, SO NH ), 7.00 (d, J = 8.4 Hz, 2H, Ar), 3.79 (s,
5
2
2
1
3
3
H, OCH3); C NMR (150 MHz, DMSO-d ) d: 158.29, 141.61, 141.07, 139.37,
6
1
27.34, 126.68, 124.69, 123.96, 123.75, 117.99, 114.34, 55.08; MS(ESI): 352.6
?
M ? Na] . Anal. Calcd for C H N O S: C 58.35 H 4.59 N 12.76, Found: C
[
1
6 15 3 3
5
8.62 H 4.18 N 12.63.
4
-(4-(3-Methoxyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide 4j Brown solid;
-
1
yield 82.6 %; m.p. 221.8–222.2 °C; IR (KBr, cm ): 3316, 3068, 1594, 1506,
466, 1405, 1344, 1226, 1166, 1103, 1029, 960, 896, 831, 786, 692, 619, 548, 459;
1
1
H NMR (600 MHz, DMSO-d ) d: 9.15 (s, 1H, pyrazole C -H), 8.34 (s, 1H,
6
3
pyrazole C -H), 8.09 (d, J = 8.4 Hz, 2H, Ar), 7.97 (d, J = 8.4 Hz, 2H, Ar), 7.44 (s,
2
5
H, SO NH ), 7.35 (d, J = 7.2 Hz, 1H, Ar), 7.33 (s, 2H, Ar), 6.85 (d, J = 7.2 Hz,
2 2
1
3
1
H, Ar), 3.83 (s, 3H, OCH3); C NMR (150 MHz, DMSO-d ) d: 159.75, 141.54,
6
1
41.25, 139,74, 132,63, 130.05, 127.35, 125.05, 124.77, 118.12, 117.76, 112.56,
?
10.79, 55.09; MS(ESI): 352.6 [M ? Na] . Anal. Calcd for C H N O S: C 58.35
1
H 4.59 N 12.76, Found: C 59.08 H 4.24 N 12.70.
1
6 15 3 3
4
-(4-(3,4-Dimethoxyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide 4k Chartreuse
-
1
solid; yield 90.8 %; m.p. 256.5–258.6 °C; IR (KBr, cm ): 3321, 3150, 3072,
2
6
8
991, 1596, 1506, 1404, 1342, 1247, 1162, 1101, 1025, 962, 900, 839, 796, 754,
1
65, 613, 547, 449; H NMR (600 MHz, DMSO-d ) d: 9.06 (s, 1H, pyrazole C -H),
6
3
.28 (s, 1H, pyrazole C -H), 8.07 (d, J = 8.4 Hz, 2H, Ar), 7.96 (d, J = 8.4 Hz, 2H,
5
Ar), 7.43 (s, 2H, SO NH ), 7.33 (d, J = 1.8 Hz, 1H, Ar), 7.27 (dd, J = 8.4 Hz,
2
2
1
J = 1.8 Hz, 2H, Ar), 7.00 (d, J = 8.4 Hz, 1H, Ar), 3.86 (s, 3H, OCH ), 3.79 (s,
2
3
1
3
3
H, OCH3); C NMR (150 MHz, DMSO-d ) d: 149.06, 147.89, 141.61, 141.05,
6
1
23

Design, synthesis and biological evaluation of some novel…
1
5
4
39.52, 127.34, 124.99, 124.16, 124.07, 117.94, 117.66, 112.18, 109.36, 55.57,
?
5.49; MS(ESI): 382.7 [M ? Na] . Anal. Calcd for C H N O S: C 56.81 H
.77 N 11.96, Found: C 57.38 H 4.41 N 11.55.
1
7 17 3 4
4
-(4-(3,4,5-Trimethoxyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide 4l Pale yel-
-
1
low solid; yield 89.1 %; m.p. 229.1–232.5 °C; IR (KBr, cm ): 3311, 3049, 1597,
504, 1403, 1363, 1333, 1247, 1159, 1126, 1004, 960, 908, 831, 777, 626, 543, 451;
1
1
H NMR (600 MHz, DMSO-d ) d: 9.13 (s, 1H, pyrazole C -H), 8.35 (s, 1H,
6
3
pyrazole C -H), 8.07 (d, J = 9.0 Hz, 2H, Ar), 7.97 (d, J = 9.0 Hz, 2H, Ar), 7.43 (s,
1
5
3
2
H, SO NH ), 7.06 (s, 2H, Ar), 3.87 (s, 6H, 2 9 OCH ), 3.68 (s, 3H, OCH );
2
C
2
3
3
NMR (150 MHz, DMSO-d ) d: 153.30, 141.55, 141.17, 139.79, 136.48, 127.35,
6
?
1
4
5
27.01, 125.09, 124.83, 118.00, 102.90, 60.09, 55.98; MS(ESI): 390.1 [M ? H]
?
28.3 [M ? K] . Anal. Calcd for C H N O S: C 55.52 H 4.92 N 10.79, Found: C
6.07 H 4.62 N 10.77.
1
8 19 3 5
Cytotoxic evaluation in vitro
Cell culture
Three different human carcinoma cell lines, MCF-7, Hela and A549, were cultured
in RPMI-1640 medium supplemented with 10 % fetal bovine serum, 100 units/mL
of penicillin, and 100 mg/mL of streptomycin. Cells were maintained at 37 °C in a
humidified atmosphere of 5 % CO in air.
2
Solutions
The compounds 4a–4l were dissolved in DMSO at a concentration of 5 mM as
stock solution, and diluted in culture medium at concentrations of 1.0, 10, 100, and
500 mM as working solutions. To avoid DMSO toxicity, the concentration of
DMSO was less than 0.1 % (v/v) in all experiments.
Cytotoxicity analysis
The cells harvested from the exponential phase were seeded equivalently into a
9
6-well plate, and then the compounds were added to the wells to achieve final
concentrations. Control wells were prepared by addition of culture medium. Wells
containing culture medium without cells were used as blanks. All experiments were
performed in quintuplicate. The MTT assay was performed as described by
Mosmann [42]. Upon completion of the incubation for 48 h, stock MTT dye
solution (20 lL, 5 mg/mL) was added to each well. After 4 h incubation,
2
-propanol (100 lL) was added to solubilize the MTT formazan. The optical
density (OD) of each well was measured on a microplate spectrophotometer at a
wavelength of 570 nm. The IC50 value was determined from plot of % viability
against dose of compounds added.
123

X. Duan et al.
Results and discussion
Chemistry
The synthetic protocol used to synthesize the title compounds (4a–4l) is outlined in
Scheme 1. 4-Hydrazinylbenzenesulfonamide hydrochloride was synthesized from
4
-aminobenzenesulfonamide by diazotization and then reduction with sodium
bisulfite in an overall yield of 44 %. 2a–2l, obtained as its perchlorate salt, were
prepared by Vilsmeyer–Haack-type reaction of substituted phenylacetic acid 1a–1l
treated with POCl /DMF. Hydrolysis of 2a–2l with sodium hydroxide aqueous
3
solution gave the key intermediates 3a–3l. The cyclization to pyrazole derivatives
bearing a benzenesulfonamide moiety was afforded by the condensation of
appropriate dialdehyde compounds 3a–3l with 4-hydrazinobenzenesulfonamide
hydrochloride in ethanol in 59–90 % yield.
The structures of 4a–4l were determined using various spectral techniques such
1
as H NMR, C NMR, FT-IR, mass spectrometry, and elemental analysis. Spectral
13
1
data IR, H NMR, C NMR and MS of compounds were found in full agreement
13
1
with the proposed structure. In H NMR spectra of these N-arylpyrazole derivatives,
two ‘‘parts’’ corresponded to the benzenesulfonamide moiety and the substituted
aromatic part of molecule were observed. The double doublets peaks at *8.08 and
*
7.97 ppm strongly suggest the phenyl ring of benzenesulfonamide. Another pair
1
of H doublets can be assigned to the para-substituted phenyl ring for most of the
obtained compounds. Two one-proton singlet peaks at 9.02–9.57 and 8.24–8.97 can
be assigned to the pyrazole ring protons. In addition, the signal for SO NH was also
2
2
1
3
observed as a two-proton singlet at d 7.43–7.48. In the C NMR spectra, aromatic
carbon resonated between d 102.9 and 161.16, while the C=N carbon resonated
around d 141.05–141.38. The IR spectra of these pyrazole derivatives displayed two
-
1
absorption bands, at 3370–3311 and 3255–3126 cm , indicative of the NH group,
in addition to the strong bands for SO N moiety at 1344–1333 and
2
2
-
166–1155 cm , and for C=N at 1601–1593 cm . The structures of the above
1
-1
1
compounds (4a–4l) were further supported by their mass spectrometry and
elemental analyses. The ESIMS of the sodium adduct of the obtained compounds
?
a–4l yielded molecular ion peaks [M ? Na] in the positive mode. Elemental
4
analysis (C, H and N) data were within ± 0.4 % of the theoretical values.
Cytotoxicity
The in vitro cytotoxic activity of compounds 4a–4l against MCF-7, Hela and A549
was evaluated by MTT assay. The IC50 values are listed in Table 1 and the activities
of celecoxib and cisplatin were used as control. The results showed that all of the
obtained compounds exhibited cytotoxicity against the tested carcinoma cell lines
with a lower IC50 value. Compound 4k, bearing the two methoxy groups on the
3-position and 4-position of the phenyl ring, was the most effective one. Its
inhibition of cell growth of MCF-7 cells was 1.2-fold higher than that found for
celecoxib under the same experimental conditions. Compound 4j with the methoxy
1
23

Design, synthesis and biological evaluation of some novel…
Table 1 The cytotoxicity of the
compounds (4a–4l) against
Compounds
IC50 (lL)
MCF-7, Hela and A549 cells
MCF-7
Hela
A549
4
4
4
4
4
4
4
4
4
4
4
4
a
b
c
d
e
f
14.46
13.25
27.26
28.85
31.09
28.51
17.75
19.23
30.98
10.89
10.40
13.54
12.34
11.06
19.30
20.96
28.72
29.02
25.99
25.82
24.22
22.72
24.86
19.36
16.95
22.74
7.76
25.52
35.00
52.62
15.97
10.04
12.01
32.12
25.79
58.94
37.54
24.48
12.02
21.84
8.53
g
h
i
j
k
l
Celecoxib
Cisplatin
8.61
group on the phenyl ring was also more toxic to MCF-7 cells than celecoxib. The
improved cytotoxic activity of 4b, 4j, 4k and 4l toward MCF-7 cells hinted that
electron-donating groups on the benzene ring could enhance the cytotoxicity of N-
arylpyrazole derivatives on comparing the activities with 4c, 4d, 4e, 4f and 4g with
electron-withdrawing groups. The position of the electron-donating group seems to
have a great effect on cytotoxic activity. Comparing compounds 4j, 4k, 4l and 4i, it
could be concluded that the substituted C’-3 position is essential for cytotoxicity
against the MCF-7 cell line.
Conclusions
In this study, a series of novel N-arylpyrazole derivatives bearing the sulfonamide
moiety were designed and synthesized, and their in vitro cytotoxicity was evaluated.
The results showed that a number of the obtained compounds exhibited potent
activity against three human cancer cell lines. Compounds 4j and 4k exhibited
better cytotoxicity than that of celecoxib and cisplatin against MCF-7. The
preliminary results show that the location and number of the methoxy group on the
benzene ring had important effects on cytotoxic activity. These results provided a
novel access to N-arylpyrazole derivatives bearing the sulfonamide moiety from
simple commercially available substituted phenylacetic acid. In addition, the
carbonic anhydrase (CA) IX activity of the obtained compounds needs to be further
studied.
Acknowledgments This work was supported by the Nature Science Fund of Hebei Province
(B2015201213, B2015201069), the Key Basic Research Special Foundation of Science Technology
123

X. Duan et al.
Ministry of Hebei Province (15962602D), the Key Research Project Foundation of Department of
Education of Hebei Province (Grant No. ZH2012041).
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