Synthesis of some new 1,3,5-trisubstituted pyrazolines bearing benzene sulfonamide as anticancer and anti-inflammatory agents

Article abstract of DOI:10.1016/j.bmcl.2011.05.061

Thirteen new 2-pyrazoline derivatives bearing benzenesulfonamide moiety (2a-m) were synthesized by condensing appropriate chalcones with 4-hydrazinonbenzenesulfonamide hydrochloride and tested for anticancer and anti-inflammatory actions. According to the

Full text of DOI:10.1016/j.bmcl.2011.05.061

Bioorganic & Medicinal Chemistry Letters 21 (2011) 4301–4305
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Synthesis of some new 1,3,5-trisubstituted pyrazolines bearing benzene
sulfonamide as anticancer and anti-inflammatory agents
Rafia Bashir a, Syed Ovais a, Shafiya Yaseen a, Hinna Hamid a, M. S. Alam a, Mohammad Samim a,
Surender Singh b, Kalim Javed a,
⇑
a Department of Chemistry, Faculty of Science, Jamia Hamdard (Hamdard University), New Delhi 110 062, India
b Department of Pharmacology, All India Institute of Medical Sciences, New Delhi 110 029, India
a r t i c l e i n f o
a b s t r a c t
Article history:
Thirteen new 2-pyrazoline derivatives bearing benzenesulfonamide moiety (2a–m) were synthesized by
condensing appropriate chalcones with 4-hydrazinonbenzenesulfonamide hydrochloride and tested for
anticancer and anti-inflammatory actions. According to the protocol of the National Cancer Institute
(NCI) in vitro disease-oriented human cells screening panel assay compounds 2b, 2c, 2e, 2f and 2g exhib-
ited considerable antitumor activities against the entire tested tumor cell lines and showed effective
Received 27 December 2010
Revised 11 May 2011
Accepted 18 May 2011
Available online 26 May 2011
growth inhibition GI50 (MG-MID) values of 2.63, 2.57, 6.61, 3.31 and 2.57
lM, respectively, beside a cyclo-
Keywords:
2-Pyrazolines
Anti-inflammatory activity
Ulcerogenic activity
Cyclooxygenase activity
Celecoxib
static activity TGI (MG-MID) 9.54, 8.51, 24.0, 19.9 and 8.71 M, respectively. Two compounds 2g and 2k
l
showed more potent anti-inflammatory activity than celecoxib at 5 h in carrageenan-induced rat paw
edema bioassay. These compounds (2g and 2k) proved to have superior gastrointestinal safety profiles
as compared to celecoxib, when tested for their ulcerogenic effects. Compounds 2g and 2k showed no
inhibition against the enzymatic activity of bovine COX-2 (in vitro).
Ó 2011 Elsevier Ltd. All rights reserved.
The sulfonamides constitute an important class of drugs, with
several types possessing a host of biological properties including
antibacterial,1 anti-carbonic anhydrase,2,3 diuretic,2,4 hypoglyce-
mic,5 antithyroid,6 and antiprotons activities.7–9 A large number
of structurally novel sulfonamide derivatives have recently been
reported to show substantial antitumor activity, both in vitro
and/or in vivo. Although they have a common chemical motif of
aromatic/heterocyclic sulfonamide, there are a variety of mecha-
nisms of their antitumor action, most of them poorly understood
at this moment. Some of these derivatives are currently being eval-
uated in clinical trials, and there is much optimism that they may
lead to novel alternative anticancer drugs, devoid of the side effects
of the presently available pharmacological agents.10 Many selec-
tive COX-2 inhibitors belong to a group of compounds which have
a central tri-substituted planar ring (five-membered heterocyclic
ring) attach to pendent benzene rings and a lipophilic group. One
of the phenyl rings is attached to SO2NH2. The SO2NH2 pharmaco-
phore is believed to induce COX-2 selectivity.11 Pyrazolines have
been reported to show a wide range of biological activity, including
anti-inflammatory,12–15 antidepressant,16 anticancer,16,17 antibac-
terial,16 antitubercular18,19 and analgesic18,19 activities.
fonamide was carried out to evaluate their anticancer and anti-
inflammatory activities.
The synthesis of novel pyrazoline derivatives has been carried
out according to the steps shown in Figure 1. In the initial step chal-
cones (1a–m) were synthesised by reacting 5-chloro-2-hydroxy-4-
methylacetophenone/5-chloro-2-methoxy-4-methylacetophenone
with appropriate aldehyde in the presence of a base by conventional
Claisen–Schmidt condensation.20 Reaction between synthesized
chalcones and 4-hydrazinobenzenesulfonamide hydrochloride in
ethanol led to synthesis of novel pyrazolines (2a–m).21 Purity of
the compounds was checked on TLC plates (silica gel G) which were
visualized by exposing to iodine vapours.
Structure of the synthesized compounds (2a–m) was established
on the basis of elemental analysis and spectral data (IR, 1H NMR, 13
C
NMR and MS). The IR spectra of the compounds 2a–m showed
absorption bands in the regions 1494–1628 cmꢀ1 corresponding to
C@N stretching bands because of ring closure. Also, in infrared spec-
tra revealed NH2 peak at 3258–3380 and 2923–3375 cmꢀ1 and for
SO2NH2 peak at 1305–1372 and 1102–1163 cmꢀ1
.
In the 1H NMR spectra of 2-pyrazolines, the three hydrogen
atoms attached to the C-4 and C-5 carbon atoms of the heterocyclic
ring gave an ABX spin system. Measured chemical shift and cou-
pling constant values (cf. Experimental Section) equivocally prove
the 2-pyrazoline structure. NH proton of the SO2NH2 group was
observed at 5.83–7.13 ppm generally as broad band. The protons
of methyl and aromatic protons were observed at expected
ppms. In the 13C NMR spectra of these compounds (2a–m), the
Due to great potential of both moieties (benzenesulfonamide
and pyrazoline), the synthesis of pyrazolines bearing benzenesul-
⇑
Corresponding author. Tel.: +91 9873463272.
E-mail address: kjaved@jamiahamdard.ac.in (K. Javed).
0960-894X/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2011.05.061

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R. Bashir et al. / Bioorg. Med. Chem. Lett. 21 (2011) 4301–4305
H3C
OR
NaOH/EtOH
Ar-CHO
+
below 10 °C
CH3
Cl
O
H3C
Cl
OR
HN NH HCl
2
H3C
H
3
OR
H
Ar
EtOH
Reflux 24h
4
+
2N
O
Ar
Cl
5
N
1
O
O
S
O
NH
2
Chalcone (1a-m)
Hydrazinobenzene
sulfonamide hydrochloride
S
O
NH
2
Pyrazoline (2a-m)
1a, 2a. R = H, Ar = phenyl
1b, 2b. R = H, Ar = 2-hydroxyphenyl
1d, 2d. R = H, Ar = 4-chlorophenyl
1f, 2f. R = H, Ar = 3,4-dimethoxyphenyl
1h, 2h. R = CH3, Ar = phenyl
1c, 2c. R = H, Ar = 2-chlorophenyl
1e, 2e. R = H, Ar = ethylenephenyl
1g, 2g. R = H, Ar = 3,4,5-trimethoxyphenyl
1i, 2i. R = CH3, Ar = 3-hydroxyphenyl
1j, 2j. R = CH3, Ar = ethylenephenyl
1k, 2k. R = CH3, Ar = N,N-dimethylaminophenyl 1l, 2l. R=CH3, Ar =3,4dimethoxyphenyl
1m, 2m. R = CH3, Ar = 3,4,5-trimethoxyphenyl
Figure 1. Synthesis of pyrazolines bearing benzenesulfonamide moiety.
chemical shift values of carbon atoms C-3 (147.88–151.65), C-4
(31.20–46.65) and C-5 (57.15–64.06).
(Table S2), beside a cyclostatic activity TGI (MG-MID) 9.54, 8.51,
24.0, 19.9 and 8.71 M, respectively (Table S3).
The ratio obtained by dividing the full panel MG-MID (
the compounds by their individual subpanel MG-MID (
considered as a measure of compound selectivity. Ratio of 3–6 re-
fer to moderate selectivity, ratios greater than six indicate high
selectivity towards the corresponding cell line, while compounds
not meeting either of these criteria are rated as nonselective.25 In
this context, the active compounds in the present study were
found to be nonselective with broad spectrum antitumor activity
against the nine tumor subpanels tested with selectivity ratios
ranges of 0.54–1.53 and 0.10–2.37 at the GI50 and TGI levels
respectively.
l
Primary in vitro one-dose (10ꢀ5 M) anticancer assay was per-
formed using full panel of about 60 human tumor cell lines in
accordance with the protocol of the Drug Evaluation Branch, Na-
tional Cancer Institute (NCI), Bethesda, and described else-
where.22–24 The human tumor cell lines were derived from nine
different cancer types: leukemia, melanoma, lung, colon, CNS,
ovarian, renal, prostate, and breast cancers. Out of the newly syn-
thesized ten compounds (2b–f, 2h and 2j–m), six compounds (2b,
2c, 2e, 2f, 2g and 2l) were selected by the National Cancer Institute
(NCI). The compound 2l displayed mild sensitivity towards pros-
trate cell line PC-3 (growth 52%) Table S1. The compounds 2b,
2c, 2e, 2f and 2g possessed considerable activity and they were se-
lected for an advanced assay against a full panel (approximately 60
l
M) of
lM) is
Table S4 presents LC50 of compounds 2b, 2c, 2e, 2f and 2g on
each cell lines while Tables S5–S7 showed mean value of GI50
cell lines) at five concentrations (100, 10, 1, 0.1 and 0.001
lM).
(MG-MID), TGI (MG-MID) and LC50.
Three response parameters (GI50, TGI, and LC50) were calculated.
The GI50 value (growth inhibitory activity) corresponds to concen-
tration of the compounds causing 50% decrease in net cell growth,
the TGI value (cytostatic activity) is the concentration of the com-
pounds resulting in total growth inhibition, and the LC50 value
(cytotoxic activity) is the concentration of the compounds causing
net 50% loss of initial cells at the end of the incubation period
(48 h). Values were calculated for each of these parameters if the
level of activity was reached; however, if the effect was not
reached or was exceeded, the value was expressed as greater or
less than the maximum or minimum concentration tested. Fur-
thermore mean graph midpoints (MG-MID) were calculated for
each of the parameters, giving an average activity parameter over
all cell lines for each compound.
Carrageenan induced hind paw edema method was used for
evaluating anti-inflammatory activity.26 Wistar rats (either sex)
weighing 150–175 g were procured from Central Animal
House facility of Jamia Hamdard, New Delhi (Registration No.
173/CPCSEA). The experiments were performed in accordance with
the guidelines for the care and use of laboratory animals, laid down
by the Committee for the Purpose of Control and Supervision of
Experiments in Animals (CPCSEA), Ministry of Social Justice and
Empowerment, Govt. of India, Jan. 2000. Overnight fasted rats
(16 h) were divided into groups of 6 animals each. One group of
rats, which served as control was given vehicle (1% CMC in water
in a volume of 10 mL/kg) only. Test compounds (20 mg/kg b.w.)
and celecoxib (20 mg/kg b.w.) suspended in vehicle (10 mL/kg)
were administered orally to respective groups. After 30 min, all
animals were injected with 0.1 mL of 1% carrageenan solution (pre-
pared in normal saline) in the subplantar aponeurosis of left hind
paw to induce inflammation and the volume of injected paw was
measured by using plethysmometer immediately (at 0 h). The
paw volume was again measured after 3 and 5 h. The average
paw volume in a group of treated rats was compared with vehicle
For the calculation of the MG-MID, insensitive cell lines are
included with the highest concentration tested.
In this Letter, 2b, 2c, 2e, 2f and 2g exhibited considerable anti-
tumor activities against the entire tested tumor cell lines. They (2b,
2c, 2e, 2f, and 2g) showed effective growth inhibition GI50
(MG-MID) values of 2.63, 2.57, 6.61, 3.31 and 2.57 lM, respectively

R. Bashir et al. / Bioorg. Med. Chem. Lett. 21 (2011) 4301–4305
4303
Table 1
Effect of 2-pyrazolines derivatives bearing benzene sulfonamide moiety on carrageenan induced hind paw edema in rats
Group No.
Treatment
Dose per kg b.w.
Increase in paw volume ml SEM after carrageenan administration
5 h
3 h
I
II
III
IV
V
VI
VII
VIII
IX
X
XI
XII
XIII
XIV
XV
Vehicle
Celecoxib
2a
2b
2c
2d
2e
2f
2g
2h
2i
2j
2k
2l
2m
10 ml/kg
20 mg
20 mg
20 mg
20 mg
20 mg
20 mg
20 mg
20 mg
20 mg
20 mg
20 mg
20 mg
20 mg
20 mg
0.38 0.0307
0.066 0.0210 (82.6%)
0.2 0.036 (47.4%)
0.42 0.0401
0.06 0.02 (85.7%)
0.166 0.056 (60.3%)
0.17 0.0422 (59.5%)
0.12 0.0167 (71.4%)
0.12 0.0167 (71.4%)
0.12 0.0307 (71.4%)
0.133 0.0494 (68.3%)
0.05 0.0342 (88.1%)
0.17 0.0333 (59.5%)
0.35 0.0224 (16.7%)
0.2 0.0365 (52.4%)
0.02 0.0167 (95.2%)
0.22 0.0703 (47.6%)
0.33 0.0558 (21.4%)
0.1 0.0365 (73.6%)
0.17 0.0333 (55.3%)
0.13 0.0333 (65.8%)
0.08 0.0307 (78.9%)
0.15 0.0429 (60.5%)
0.12 0.0307 (68.4%)
0.17 0.0211 (55.3%)
0.23 0.0615 (39.5%)
0.17 0.0494 (55.3%)
0.08 0.0167 (78.9%)
0.26 0.0422 (31.6%)
0.25 0.0342 (34.2%)
⁄P <0.05 compared to control (one-way ANOVA followed by Dunnett’s test). Values are presented as mean SEM (n = 6). Values in parentheses
represent percent inhibitions.
(control group) and the percentage inhibition of edema was calcu-
lated by using the formula:
cell lines. The results of this assay indicated an activity in the
micro-molar ranged linked particularly five compounds (2b, 2c,
2e, 2f and 2g). The synthesized compounds were also evaluated
for their anti-inflammatory activity in carrageenan-induced rat
paw edema bioassay. Two compounds 2g and 2k showed more po-
tent anti-inflammatory activity than celecoxib at 5 h. These com-
pounds (2g and 2k) proved to have superior gastrointestinal
safety profiles as compared to celecoxib, when tested for their
ulcerogenic effects. Compounds 2g and 2k showed no inhibition
against the enzymatic activity of bovine COX-2 (in vitro).
Percent inhibition ¼ ð1 ꢀ Vt=VcÞ ꢁ 100
where Vt is the mean paw volume of the test drug treated rats and
Vc is the mean paw volume of the control.
The results (Table 1) show that all compounds except 2i, 2l and
2m caused more than 50% reduction of oedema at 3 or 5 h. the
most pronounced anti-inflammatory activity among the com-
pounds studied was observed for 2g and 2k. The efficacy of 2g
and 2k was higher than that of celecoxib at 5 h.
Structure–activity relationship showed that introduction of
electron donating groups in 5-aryl ring of pyrazolines (2a–g) syn-
thesised from appropriate 20-hydroxychalcone (1a–g) enhanced
the anti-inflammatory activity. On the other hand, introduction
of electron donating group in the 5-aryl ring of pyrazolines
(2h–m) synthesised from appropriate 20-methoxy chalcones
(1h–m) reduced the activity.
Acute gastric ulcerogenic effect of the compound 2g and 2k
were evaluated in Wistar rats.27 Albino rats of Wistar strain
(160–220 g) fasted over 24 h were randomly allotted into three
groups of six animals each. The animals of one group were given
vehicle 10 mL/kg (CMC 1% w/v in distilled water) orally. The com-
pound 2g (60 mg/kg) and 2k (60 mg/kg) suspended in vehicle were
administered orally in a volume of 10 mL/kg to the animals of two
different groups, respectively. They were scarified under deep
ether anaesthesia after six hours of the treatment at a dose of
60 mg/kg (three times). Their stomach were removed and opened
through greater curvature for examining lesions or bleedings.
Compounds 2g and 2k showed no ulcerogenic activity.
Acknowledgments
The authors are very grateful to the staff members of Depart-
ment of Health and Human Services, National Cancer Institute
(NCI), Bethesda, Maryland, USA, for carrying out in vitro anti cancer
screening of the newly synthesized compounds. This work was
supported by Grant No. 32-228/2006 (SR) from the University
Grants Commission, New Delhi, India. One of the authors, Rafia
Bashir is thankful to UGC for fellowship. Thanks are due to SAIF,
CDRI, Lucknow, for providing Mass spectra.
Supplementary data
Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.bmcl.2011.05.061.
References and notes
The compounds (2g and 2k) were also tested for their ability to
inhibit bovine COX-1 and COX-2 enzymes in vitro. Cyclooxygenase
activity was assayed using COX inhibitory screening kit (Cayman
chemicals) by the method of Gierse et al.28 These compounds were
found fail to inhibit the activity of COX-2. However, they (2g and
2k) showed very mild inhibition against the enzymatic activity of
COX-1. On the basis of these observations it may be assumed that
these derivatives inhibit enzymatic activity of 5-lipo-oxygenase
which like COX-2 is responsible for modification of arachidionic
acid to produce proinflammatory molecules prostanoids and
leukotrienes.
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In conclusion, the present study describes the synthesis of
pyrazolines bearing benzene sulfonamide moiety (2a–m). The
cytotoxicity of six compounds (2b, 2c, 2e, 2f, 2g and 2l) was
evaluated in vitro in a screen panel consisting of 60 human tumor

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phenyl protons), 7.57 (1H, s, H-60), 7.67 (2H, d, 7.82 Hz, N-phenyl protons),
10.30 (1H, s, OH-20); FAB-MS (m/z): 467 [M+, base peak], 466 [M+ꢀ1], 387
[M+ꢀSO2NH2], 157; 13C NMR (DMSO, d): 20.22 (–CH3), 41.48 (C-4 pyrazoline),
60.89 (C-5 pyrazoline), 128.15 (C-a), 134.05 (C-b), 151.65 (C-3 pyrazoline),
155.31 (C-20). Anal. Calcd for C24H22CIN3O3S: C, 61.60; H, 4.74; N, 8.98; S, 6.85.
Found: C, 61.54; H, 4.70; N, 8.97; S, 6.83.
3-(50-Chloro-20-hydroxy-40-methylphenyl)-5-(3,4-dimethoxyphenyl)-1-(p-
sulfamylphenyl)-
mp 229–231 °C; Rf = 0.26, (Petrol/acetone 7:3). IR
D
2-pyrazoline (2f): Recrystallized from methanol; yield = 60%;
m
max (solvent, in cmꢀ1): 3253
19. Chandra, T.; Garg, N.; Lata, S.; Saxena, K. K.; Kumar, A. Eur. J. Med. Chem. 2010,
45, 1772.
(OH), 1592 (C@N), 1510, 1330 and 1155 cmꢀ1 (SO2N<); 1H NMR (400 MHz,
DMSO, d): 2.26 (3H, s, CH3-40), 3.65 (3H, s, OCH3-3), 3.68 (3H, s, OCH3-4), 3.97
[1H, dd, J = 12.40 Hz, 18.00 Hz, H-4 cis (pyrazoline)], 5.47 [1H, dd, J = 6.40 Hz,
J = 10.80 Hz, H-5 (pyrazoline)], 6.64 (1H, d, J = 8.00 Hz, H-5), 6.83 (1H, d,
J = 7.6 Hz, H-6), 6.87 (2H, d, J = 8.80 Hz, H-300, H-500), 6.90–7.02 (4H, m, H-2, H-
20, SO2NH2), 7.54 (2H, d, J = 8.80 Hz, H-200, H-600), 7.57 (1H, s, H-60), 10.29 (1H, s,
OH); FAB-MS (m/z): 501 [M+, base peak], 500 [Mꢀ1], 437 [M+ꢀSO2], 421
[M+ꢀSO2NH2], 364 [M+ꢀ5-aryl ring]; 13C NMR (DMSO, d): 20.19 (–CH3), 44.79
(C-4 pyrazoline), 55.92 (OCH3 at C-3, C-4), 62.03 (C-5 pyrazoline), 150.78 (C-3
pyrazoline), 155.28 (C-20). Anal. Calcd for C24H24CIN3O5S: C, 57.42; H, 4.82; N,
8.37; S, 6.39. Found: C, 57.37; H, 4.79; N, 8.36; S, 6.37.
20. General method for the synthesis of chalcones (1a–m): To a cold solution
(below 10 °C) of acetophenone (0.01 mol) and desired aromatic aldehyde
(0.01 mol) in ethanol (20 ml) was added a chilled solution of sodium hydroxide
(10 ml of 30% solution). The reaction mixture was covered with a layer of
petroleum ether (60–80 °C) and left at room temperature for a period of 12 h. It
was diluted with water (100 ml) and acidified with concd HCl. The product
obtained was filtered, washed with water and crystallized from ethanol.
21. General method for the synthesis of 3,5-diaryl-1-(p-sulfamylphenyl)-2-pyrazolines
(2a–m): Appropriate chalcones (0.001 mol) and 4-hydrazinobenzenesulf-
onamide hydrochloride (0.001 mol) were dissolved in ethanol (25 ml) and
refluxed for 24 h. After completion of the reaction, refluxing condenser was
removed and the reaction solution was concentrated to 10–15 ml. It was left at
room temperature to give crystalline compound. It was filtered and
3-(50-Chloro-20-hydroxy-40-methylphenyl)-5-(3,4,5-trimethoxyphenyl)-1-(p-
sulfamylphenyl)-D
2-pyrazoline (2g): Recrystallized from acetone, yield = 64%;
mp 237–238 °C; Rf = 0.75, (petroleum ether (60–80)/toluene/ethyl acetate;
10:5:3); IR mmax (solvent, in cmꢀ1): 3409 (OH), 2923 NH2, 1592 (C@N), 1502,
1328 and 1124 cmꢀ1 (SO2N<); 1H NMR (400 MHz, DMSO, d): 2.27 (3H, s, CH3-
40), 3.59 (3H, s, OCH3), 3.65 (6H, s, OCH3), 4.05 [1H, m, H-4 trans (pyrazoline)],
5.42 [1H, dd, J = 5.6 Hz, J = 10.32 Hz, H-5 (pyrazoline)], 6.56 (2H, s, SO2NH2),
6.87 (1H, s, H-30), 6.94–7.04 (4H, m, H-300, H-500, H-2, H-6), 7.54 (1H, s, H-60),
7.62 (2H, d, H-200, H-600), 10.27 (1H, s, OH); FAB-MS (m/z): 531 [M+, base peak],
530 [M+ꢀ1], 467 [M+ꢀSO2], 451 [M+ꢀSO2NH2], 364 [M+ꢀ5-aryl ring]; 13C NMR
(DMSO, d): 20.15 (–CH3), 44.89 (C-4 pyrazoline), 56.34 (OCH3 at C-3, C-4),
60.41 (OCH3 at C-5), 62.71 (C-5 pyrazoline), 150.91 (C-3 pyrazoline), 155.34 (C-
20). Anal. Calcd for C25H26CIN3O6S: C, 56.44; H, 4.93; N, 7.90; S, 6.03. Found: C,
56.39; H, 4.88; N, 7.89; S, 6.01.
recrystallised to give pure compound. For preparing pyrazoline
amount of NaOAc was added to refluxing alcohol.
a small
3-(50-Chloro-20-hydroxy-40-methylphenyl)-5-phenyl-1-(p-sulfamylphenyl)-D2
-
pyrazoline (2a): Recrystallized from methanol; yield = 48%; mp 217–219 °C;
Rf = 0.57; (toluene/ethyl acetate/formic acid, 5:4:1). IR
m
max (solvent, in cmꢀ1):
3376 (OH), 3323 and 3269 (NH2), 1599 (C@N), 1305 and 1150 cmꢀ1 (SO2N<);
1H NMR (200 MHz, DMSO, d): 2.31 (3H, s, CH3), 2.76 (1H, dd, J = 12.14 Hz,
J = 12 Hz, H-4 cis (pyrazoline)], 5.25 [1H, d, J = 9.4 Hz, H-5 (pyrazoline)], 7.01
(1H, s, H-30), 7.11 (2H, s, SO2NH2), 7.32 (2H, d, J = 8.8 Hz, H-300, H-500), 7.40–7.59
(5H, m, H-2, H-3, H-4, H-5, H-6), 7.68 (2H, d, J = 8.8 Hz, H-200, H-600), 8.00 (1H, s,
H-30), 9.89 (1H, s, OH); MALDI (m/z): 441 [M+], 442 [M+1], 444, 464 [M+Na].
Anal. Calcd for C22H20CIN3O3S: C, 59.79; H, 4.56; N, 9.51; S, 7.26. Found: C,
59.74; H, 4.53; N, 9.50; S, 7.24.
3-(50-Chloro-20-methoxy-40-methylphenyl)-5-(phenyl)-1-(p-sulfamylphenyl)-D2
pyrazoline (2h): Recrystallized from methanol, yield = 50%; mp 132–134 °C;
Rf = 0.79, (toluene/ethyl acetate/formic acid, 5:4:1); IR
max (solvent, in cmꢀ1):
-
m
3-(50-Chloro-20-hydroxy-40-methylphenyl)-5-(2-hydroxyphenyl)-1-(p-
3385 & 3261 (NH2), 1591 (C@N), 1506, 1370 and 1154 cmꢀ1 (SO2N<); 1H NMR
(300 MHz, DMSO, d): 2.33 (3H, s, CH3), 3.25 [1H, dd, J = 6.20 Hz, J = 18.00 Hz, H-
4 (trans)], 4.01 [1H, dd J = 12.00 Hz, J = 18.10 Hz, H-4 (cis)], 3.77 (3H, s, OCH3),
5.55 [1H, dd, J = 5.1 Hz, J = 12.00 Hz, H-5], 7.10 (1H, s, H-30), 7.03–7.23 (5H, 2 m,
phenyl protons), 7.32 (2H, d, J = 8.7 Hz, H-200, H-600), 7.54 (2H, d, J = 8.7 Hz, H-300,
H-500), 7.88 (1H, s, H-60); MALDI (m/z): 455 [M+], 454 [Mꢀ1], 457 [M+2], 376;
13C NMR (DMSO, d): 20.39 (–CH3), 46.48 (C-4 pyrazoline), 56.56 (OCH3 at C-20),
62.77 (C-5 pyrazoline), 147.89 (C-3 pyrazoline), 156.50 (C-20). Anal. Calcd for
sulfamylphenyl)-D
2-pyrazoline (2b): Recrystallized from methanol; yield = 40%;
mp 212–213 °C; Rf = 0.67, (toluene/ethyl acetate/formic acid, 5:4:1); IR mmax
(solvent, in cmꢀ1): 3260 (OH), 1599 (C@N), 1338 and 1155 cmꢀ1 (SO2N<); 1H
NMR (400 MHz, DMSO, d): 3.11 [1H, m, H-4 trans (pyrazoline)], 3.84 (3H, s,
CH3), 3.99 [3H, m, H-4 cis (pyrazoline)], 5.64 [3H, m, H-5 (pyrazoline)], 6.65
(1H, d, J = 7.20 Hz, H-3), 6.83 (2H, d, J = 8.00 Hz, H-300, H-500), 6.92–7.04 (6H, m,
H-30, H-4, H-5, H-60, SO2NH2), 7.53–7.57 (3H, m, H-6, H-200, H-600), 9.92 (1H, s,
OH), 10.32 (1H, s, OH); FAB-MS (m/z): 457 [M+, base peak], 456 [M+ꢀ1], 393
[M+ꢀSO2], 377 [M+ꢀSO2NH2], 364 [M+ꢀ5-aryl ring], 271; 13C NMR (DMSO, d):
20.20 (–CH3), 43.37 (C-4 pyrazoline), 57.15 (C-5 (yrazoline), 148.39 (C-3
pyrazoline), 154.68 (C-20). Anal. Calcd for C22H20CIN3O4S: C, 57.70; H, 4.40; N,
9.18; S, 7.00. Found: C, 57.68; H, 3.93; N, 9.17; S, 6.99.
C
24H25N3O3S: C, 60.59; H, 4.86; N, 9.22; S, 7.03. Found: C, 60.53; H, 4.82; N,
9.21; S, 7.02.
3-(50-Chloro-20-methoxy-40-methylphenyl)-5-(3-hydroxyphenyl)-1-(p-
sulfamylphenyl)-D
2-pyrazoline (2i): Recrystallized from methanol; yield = 50%;
mp 216–218 °C; Rf = 0.41, (toluene/ethyl acetate/formic acid, 7.5:2:0.5); IR mmax
3-(50-Chloro-20-hydroxy-40-methylphenyl)-5-(2-chlorophenyl)-1-(p-
(solvent, in cmꢀ1): 3359 (NH2), 1591 (C@N), 1506, 1370 and 1152 cmꢀ1
1
sulfamylphenyl)-
D
2-pyrazoline (2c): Recrystallized from methanol; yield = 30%;
(SO2N<); H NMR (300 MHz, DMSO, d): 2.36 (3H, s, CH3-40), 3.79 (3H, s,
mp 221 °C; Rf = 0.71, (toluene/ethyl acetate/formic acid, 5:4:1); IR mmax
(solvent, in cmꢀ1): 3364 (OH), 3317 & 3258 (NH2), 1596 (C@N), 1311 &
1102 cmꢀ1 (SO2N<); 1H NMR (400 MHz, DMSO, d): 2.28 (3H, s, CH3), 2.70 [1H,
m, H-4 trans (pyrazoline)], 5.43 [1H, m, H-5 (pyrazoline)], 6.99 (1H, s, H-30),
7.06 (2H, s, SO2NH2), 7.28 (2H, d, J = 7.6 Hz, H-300, H-500), 7.44 (2H, m, H-4, H-5),
7.52 (1H, d, J = 6.4 Hz, H-3), 7.64 (2H, d, J = 7.6 Hz, H-200, H-600), 7.72 (1H, d, H-6,
J = 6.4 Hz), 7.98 (1H, s, H-60), 9.86 (1H, s, OH); FAB-MS (m/z): 475 [M+], 474
[M+ꢀ1], 411 [M+ꢀSO2], 395 [M+ꢀSO2NH2], 364 [M+ꢀ5-aryl ring], 77 (base
peak); 13C NMR (DMSO, d): 20.10 (–CH3), 31.20 (C-4 pyrazoline), 74.17 (C-5
pyrazoline), 148.39 (C-3 pyrazoline), 154.68 (C-20). Anal. Calcd for
OCH320), 3.97 [1H, m, H-4 trans (pyrazoline)], 5.45 [1H, m, H-5 (pyrazoline)],
6.58–6.70 (3H, m, H-30, H-4, H-5), 7.03–7.13 (6H, m, H-6, H-2, H-300, H-500,
SO2NH2), 7.58 (2H, d, H-200,H-600), 7.89 (1H, s, H-60), 9.47 (1H, s, OH-3); FAB-MS
(m/z): 470 [M+], 471 [M++1], 407 [M+ꢀSO2], 391 [M+ꢀSO2NH2], 378 [M+ꢀ5-aryl
ring], 157; 13C NMR (DMSO, d): 20.20 (–CH3), 46.43 (C-4 pyrazoline), 56.53
(OCH3 at C-20), 62.72 (C-5 pyrazoline), 147.91 (C-3 pyrazoline), 158.34 (C-20).
Anal. Calcd for C23H22ClN3O4S: C, 58.53; H, 4.70; N, 8.90; S, 6.79. Found: C,
58.48; H, 4.67; N, 8.90; S, 6.78.
3-(50-Chloro-20-methoxy-40-methylphenyl)-5-(ethylenephenyl)-1-(p-
sulfamylphenyl)-
D
2-pyrazoline
(2j):
Recrystallized
from
methanol;
C
22H19CI2N3O3S: C, 55.47; H, 4.02; N, 8.82; S, 6.73. Found: C, 55.41; H, 3.99;
yield = 53.6%; mp 222–226 °C; Rf = 0.27, (toluene/ethyl acetate/formic acid,
7.5:2:0.5); IR mmax (solvent, in cmꢀ1): 3258 (NH2), 1590 (C@N), 1370,
1096 cmꢀ1 (OCH3) and 1153 cmꢀ1 (SO2N<); 1H NMR (300 MHz, DMSO, d):
2.35 (3H, s, CH3), 3.31 [1H, d, H-4 trans (pyrazoline)], 3.75 [1H, d, H-4 cis
N, 8.82; S, 6.71.
3-(50-Chloro-20-hydroxy-40-methylphenyl)-5-(4-chlorophenyl)-1-(p-
sulfamylphenyl)-
and methanol; yield = 34%; mp 236–238 °C; Rf = 0.41, (toluene/ethyl acetate/
formic acid, 5:4:1); IR
max (solvent, in cmꢀ1): 3450 (OH), 3326 and 3375 (NH2),
D
2-pyrazoline (2d): Recrystallized from the mixture of acetone
(pyrazoline)], 3.82 (3H, s, OCH3), 5.15 (1H, m, H-5), 6.30 (1H, d, H-a), 6.65 (1H,
m
d, H-b), 7.02 (2H, s, SO2NH2), 7.12 (1H, s, H-30), 7.22–7.28 (5H, m, cinnamal unit
protons), 7.40 (2H, d, J = 8.6 Hz, H-300, H-500), 7.62 (2H, d, J = 8.7 Hz, H-200, H-600),
7.83 (1H, s H-60); FAB-MS (m/z): 481[M+], 482 [M++1, base peak], 401[M+ꢀ80],
157; 13C NMR (DMSO, d): 20.39 (–CH3), 43.36 (C-4 pyrazoline), 56.59 (OCH3 at
1595 (C@N), 1319 and 1136 cmꢀ1 (SO2N<); 1H NMR (300 MHz, DMSO, d): 2.31
(3H, s, CH3-40), 2.75 [1H, dd, J = 12.14 Hz, J = 16.74 Hz, H-4 cis (pyrazoline)],
5.28 [1H, d, H-5 (pyrazoline)], 7.02 (1H, s, H-30), 7.10 (2H, s, SO2NH2), 7.33 (2H,
d, J = 8.51 Hz, H-300, H-500), 7.53 (2H, d, J = 8.27 Hz, H-3, H-5), 7.60 (2H, d,
J = 8.29 Hz, H-2, H-6), 7.68 (2H, d, J = 8.51 Hz, H-200, H-600), 7.99 (1H, s, H-60),
9.90 (1H, s, OH); FAB-MS (m/z): 476 [M++1], 475 [M+], 411 [M+ꢀSO2], 395
[M+ꢀSO2NH2], 364 [M+ꢀ5-aryl ring], 157; 13C NMR (DMSO, d): 20.09 (–CH3),
32.19 (C-4 pyrazoline), 76.29 (C-5 pyrazoline), 148.45 (C-3 pyrazoline), 154.66
(C-20). Anal. Calcd for C22H19 CI2 N3O3S; C, 55.47; H, 4.02; N, 8.82; S, 6.73.
Found: C, 55.42; H, 3.99; N, 8.81; S, 6.72.
C-20), 61.64 (C-5 pyrazoline), 127.86 (C-
a), 136.34 (C-b), 148.73 (C-3
pyrazoline). Anal. Calcd for C25H24ClN3O3S: C, 62.30; H, 5.02; N, 8.72; S, 6.65.
Found: C, 62.24; H, 4.97; N, 8.71; S, 6.64.
3-(50-Chloro-20-methoxy-40-methylphenyl)-5-(4-(N,N-dimethylamino)-phenyl)-1-
(p-sulfamylphenyl)-D2-pyrazoline (2k): Recrystallized from methanol;
yield = 74.5%; mp 210–211 °C; Rf = 0.67 (toluene/ethyl acetate/formic acid,
5:4:1); IR mmax (solvent, in cmꢀ1): 3380 (NH2), 1591 (C@N), 1506, 1367 and
1153 cmꢀ1 (SO2N<); 1H NMR (300 MHz, DMSO, d): 2.34 (3H, s, CH3-40), 2.86
[6H, s, N(CH3)2], 3.15 [1H, m, H-4 trans (pyrazoline)], 3.77 (3H, s, OCH3), 3.92
[1H, m, H-4 cis (pyrazoline)], 5.44 [1H, m, H-5 (pyrazoline)], 6.78 (2H, d,
J = 8.40 Hz, H-3, H-5), 7.03–7.09 (7H, m, five aromatic protons and SO2NH2),
7.55 (2H, d, H-2, H-6, J = 8.67 Hz), 7.87 (1H, s, H-60); FAB-MS (m/z): 498 [M+,
base peak], 497 [M+ꢀ1], 434 [M+ꢀSO2], 418 [M+ꢀSO2NH2], 378 [M+ꢀ5-aryl
ring]; 13C NMR (DMSO, d): 20.39 (CH3), 41.96 (C-4 pyrazoline)), 46.49
(N(CH3)2), 56.55 (OCH3), 62.35 (C-5 pyrazoline), 147.88 (C-3 pyrazoline),
3-(50-Chloro-20-hydroxy-40-methylphenyl)-5-(ethylenephenyl)-1-(p-
sulfamylphenyl)-D
2-pyrazoline (2e): Recrystallized from acetone; yield = 64%;
mp 267–268 °C; Rf = 0.47, (toluene/ethyl acetate/formic acid, 7.5:2:0.5); IR mmax
(solvent, in cmꢀ1): 3451 (OH), 3323 & 3342 (NH2), 1628 (C@N), 1327 and
1144 cmꢀ1 (SO2N<); H NMR (300 MHz, DMSO, d): 2.31 (3H, s, CH3-40), 3.85
1
[1H, m, H-4 cis (pyrazoline)], 5.23 [1H, m, H-5 (pyrazoline)], 6.32 (1H, q, –
CH@CH), 6.73 (1H, d, J = 15.82 Hz, –CH@CH–), 6.99 (1H, s, H-30), 7.07 (2H, s,
SO2NH2), 7.18–7.30 (5H, m, H-2, H-3, H-4, H-5, H-6), 7.42 (2H, d, J = 7.82 Hz, N-

R. Bashir et al. / Bioorg. Med. Chem. Lett. 21 (2011) 4301–4305
4305
156.48 (C-20). Anal. Calcd for C25H27Cl N4O3S: C, 60.17; H, 5.45; N, 11.23; S,
6.43. Found: C, 60.11; H, 5.41; N, 11.22; S, 6.40.
CH3-40), 3.32 [1H, dd, J = 12.13 Hz, J = 18.29 Hz, H-4 trans (pyrazoline)], 3.81
(12H, s, 4 ꢁ OCH3), 4.01 [1H, dd, J = 6.31 Hz, J = 11.92 Hz, H-4 cis (pyrazoline)],
5.19 (1H, m, H-5 pyrazoline), 6.28 (2H, s, SO2NH2), 6.47 (2H, s, H-2, H-6), 6.82
(1H, s, H-30), 7.06 (2H, d, J = 8.22 Hz, H-300, H-500), 7.69 (2H, d, J = 8.23 Hz, H-200,
H-600), 7.97 (1H, s, H-60); FAB-MS (m/z): 545 [M+, base peak], 544 [M+ꢀ1], 481
[M+ꢀSO2], 465 [M+ꢀSO2NH2], 157; 13C NMR (DMSO, d): 20.50 (–CH3), 46.65 (C-
4 pyrazoline), 55.85 (OCH3 at C-20), 56.13 (OCH3 at C-3, C-5), 60.63 (OCH3 at C-
4), 64.06 (C-5 pyrazoline), 147.99 (C-3 pyrazoline), 156.10 (C-20). Anal. Calcd
for C26H28Cl N3O6S: C, 57.19; H, 5.17; N, 7.70; S, 5.87. Found: C, 57.13; H, 5.12;
N, 7.69; S, 5.86.
3-(50-Chloro-20-methoxy-40-methylphenyl)-5-(3,4-dimethoxyphenyl)-1-(p-
sulfamylphenyl)-D
2-pyrazoline (2l): Recrystallized from methanol, yield = 61%;
mp 223–225 °C; Rf = 0.57 (toluene/ethyl acetate/formic acid, 5:4:1); IR mmax
(solvent, in cmꢀ1): 3335 and 3342 (NH2), 1588 (C@N), 1506, 1368 cmꢀ1 and
1
1152 cmꢀ1 (SO2N<); H NMR (300 MHz, DMSO, d): 2.40 (3H, s, CH3-40), 3.81
(3H, s, OCH3), 3.34 [1H, dd, J = 6.21 Hz, J = 18.22 Hz, H-4 trans (pyrazoline)],
3.82 (3H, s, OCH3), 3.85 (3H, s, OCH3-40), 3.97 [1H, dd, J = 12.36 Hz, J = 18.36 Hz,
H-4 cis (pyrazoline)], 5.23 [1H, dd, J = 6.05 Hz, J = 11.57 Hz, H-5 (pyrazoline),
5.83 (2H, s, SO2NH2), 6.74–6.82 (4H, m, H-30, H-2, H-5, H-6), 7.08 (2H, d,
J = 8.56 Hz, H-300, H-500), 7.70 (2H, d, J = 8.57 Hz, H-200, H-600), 7.97 (1H, s, H-60);
FAB-MS (m/z): 515 [M+], 516 [M++1, base peak], 451 [M+ꢀSO2], 435
[M+ꢀSO2NH2], 378 [M+ꢀ5-aryl ring], 157; 13C NMR (DMSO, d): 20.39 (–CH3),
46.54 (C-4 pyrazoline), 55.88 (OCH3 at C-20), 55.92 (OCH3 at C-3), 56.57 (OCH3
at C-4), 62.78 (C-5 pyrazoline), 149.50 (C-3 pyrazoline), 149.50 (C-20). Anal.
Calcd for C25H26ClN3O5S: C, 58.19; H, 5.08; N, 8.14; S, 6.21. Found: C, 58.13; H,
5.04; N, 8.14; S, 6.20.
22. Monks, A.; Scudiero, D.; Skehan, P.; Shoemaker, R.; Paull, K.; Vistica, D.;
Hose, C.; Langley, J.; Cronise, P.; Vaigro-Wolf, A. J. Natl. Cancer Inst. 1991,
83, 57.
23. Boyd, M. R.; Paull, K. D. Drug Dev. Res. 1995, 34, 91.
24. Boyd, M. R. In Cancer Drug Discovery and Development: Anticancer Drug
Development Guide: Preclinical Screening, Clinical Trials, and Approval; Teicher,
B. A., Ed.; Humana Press: Totowa, New Jersey, 1997; pp 23–43. Chapter 2.
25. Acton, E. M.; Narayanan, V. L.; Risbood, P. A.; Shoemaker, R. H.; Vistica, D. T.;
Boyd, M. R. J. Med. Chem. 1994, 37, 2185.
26. Winter, C. A.; Risley, E. A.; Nuss, G. W. Proc. Soc. Exp. Biol. Med. 1962, 111, 544.
27. Daidone, G.; Maggio, B.; Raffa, D. Eur. J. Med. Chem. 1994, 29, 707.
28. Gierse, J. K.; Koboldt, C. M.; Walker, M. C.; Siebeil, K.; Isakson, P. C. Biochem. J.
1999, 339, 607.
3-(50-Chloro-20-methoxy-40-methylphenyl)-5-(3,4,5-trimethoxyphenyl)-1-(p-
sulfamylphenyl)-
D
2-pyrazoline
(2m):
Recrystallized
from
methanol;
yield = 67%; mp 239–241 °C; Rf = 0.60 (toluene/ethyl acetate/formic acid,
5:4:1); IR mmax (solvent, in cmꢀ1): 3374 (NH2), 3269, 1591 (C@N), 1363 and
1152 (SO2N<), 1097 cmꢀ1 (OCH3); 1H NMR (300 MHz, DMSO, d): 2.40 (3H, s,