Synthesis of some sulfonamides, disubstituted sulfonylureas or thioureas and some structurally related variants. A class of promising antitumor agents

Article abstract of DOI:10.1007/s00044-007-9033-8

Some new benzenesulfonamides, disubstituted sulfonylureas, and sulfonylthioureas substituted basically with 3-(2-thienyl or 3-pyridyl)-indeno[1,2-c]pyrazol(in)e counterpart were synthesized to be evaluated for their in vitro antitumor activity. Some of the thioureido derivatives were cyclized to the corresponding five-membered thiazolidinons, thiazolines, and the six-membered thiazinones as interesting structure variants. According to the protocol of the National Cancer Institute (NCI) in vitro disease-oriented human cells screening panel assay, 13 compounds showed promising broad spectrum antitumor activity. In general, compounds containing the thienyl moiety displayed better antitumor spectra than those containing the pyridyl moiety. Compound 5, 4-(3-(2-thienyl)-3H-indeno[1,2-c]pyrazol-2-yl)- benzenesulfonamide [GI₅₀, TGI, and LC₅₀ (MG-MID) values of 13.2, 33.1 and 69.2 μM, respectively] proved to be the most active member in this study with variable degrees of potencies against all the tested subpanel tumor cell lines and particular effectiveness against the leukemia and prostate subpanels at both the GI₅₀ (3.30 and 8.68 μM, respectively) and the TGI levels (15.7 and 22.3 μM, respectively).

Full text of DOI:10.1007/s00044-007-9033-8

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res (2007) 16:300–318
DOI 10.1007/s00044-007-9033-8
ORIGINAL RESEARCH
Synthesis of some sulfonamides, disubstituted
sulfonylureas or thioureas and some structurally
related variants. A class of promising antitumor agents
Hassan M. Faidallah Æ Mohammed S. Al-Saadi Æ
Sherif A. F. Rostom Æ Hesham T. Y. Fahmy
Received: 8 May 2007 / Accepted: 24 September 2007 / Published online: 30 October 2007
¨
Ó Birkhauser Boston 2007
Abstract Some new benzenesulfonamides, disubstituted sulfonylureas, and
sulfonylthioureas substituted basically with 3-(2-thienyl or 3-pyridyl)-indeno
[1,2-c]pyrazol(in)e counterpart were synthesized to be evaluated for their in vitro
antitumor activity. Some of the thioureido derivatives were cyclized to the corre-
sponding five-membered thiazolidinons, thiazolines, and the six-membered
thiazinones as interesting structure variants. According to the protocol of the
National Cancer Institute (NCI) in vitro disease-oriented human cells screening
panel assay, 13 compounds showed promising broad spectrum antitumor activity. In
general, compounds containing the thienyl moiety displayed better antitumor
spectra than those containing the pyridyl moiety. Compound 5, 4-(3-(2-thienyl)-3H-
indeno[1,2-c]pyrazol-2-yl)-benzenesulfonamide [GI50, TGI, and LC50 (MG-MID)
values of 13.2, 33.1 and 69.2 lM, respectively] proved to be the most active
member in this study with variable degrees of potencies against all the tested
subpanel tumor cell lines and particular effectiveness against the leukemia and
prostate subpanels at both the GI50 (3.30 and 8.68 lM, respectively) and the TGI
levels (15.7 and 22.3 lM, respectively).
The wide spread of cancer in recent years has prompted the search for new
therapeutic agents in this field. As part of our ongoing program aimed towards the
development of new potential chemotherapeutic agents (Fahmy et al., 2002 and
2003; Bekhit et al., 2003; Rostom et al., 2003; Al-Saadi et al., 2005), the synthesis
H. M. Faidallah Á M. S. Al-Saadi Á S. A. F. Rostom
Department of Medicinal Chemistry, Faculty of Medicine, King Abdulaziz University,
P.O. Box 80205, Jeddah 21589, Saudi Arabia
H. T. Y. Fahmy (&)
Department of Pharmaceutical Sciences, College of Pharmacy, South Dakota State University,
Box 2202C, Brookings, SD 57007, USA
e-mail: Hesham.Fahmy@sdstate.edu

Med Chem Res (2007) 16:300–318
301
and antitumor evaluation of a series of 3-(4-chlorophenyl)-indeno[1,2-c]-pyra-
zol(in)es substituted with benzenesulfonamide, disubstituted sulfonylurea,
sulfonylthiourea pharmacophores, and some derived thiazolidinone and thiazoline
ring systems were reported (A and B, Fig. 1) (Rostom, 2006). This investigation
resulted in eight compounds with broad spectrum antitumor activity according to the
protocol of the National Cancer Institute (NCI) in vitro disease-oriented human cells
screening panel assay. The data obtained revealed that the oxidized pyrazole
derivatives exhibited better antitumor activity than the parent pyrazoline analogs,
whereas the benzenesulfonamides and N1,N3-disubstituted sulfonylureas showed
significant antitumor spectrum better than that of the sulfonylthioureido and derived
thiazole analogs (Rostom, 2006).
Interest in this class of compounds stemmed from the well-documented
antitumor activity displayed by some derivatives containing the aforementioned
functionalities (Mohamadi et al., 1992; Toth et al., 1997; Medina et al., 1998 and
1999; Lee et al., 2002; Chern et al., 1997), including Sulofenur (LY 186641) C and
its structural analogs (LY 295501) D and E (Fig. 1) (Mastrolorenzo et al., 2000).
These compounds (C–E) were shown to strongly inhibit the growth of several
tumors through various mechanisms of action (Mastrolorenzo et al., 2000;
Scozzafava and Supuran, 2000; Casini et al., 2002; De Simone et al., 2006),
however, with a multitude of side-effects (Guan et al., 2002). In addition, pyrazole-
containing compounds have been reported to contribute to a vareity of anticancer
activities (Daidone et al., 2004; Brough et al., 2005; Krystoet et al., 2006; Huang
et al., 2007). These facts prompted the continuation of study of this class of
compounds (A, B; Fig. 1); another series of analogs was synthesized in which the
indeno[1,2-c]-pyrazol(in)e counterpart, sulfonamide, sulfonylurea, and thiourea
R
H
Y
N
N
X
R
H
N
N
N
N
S
SO2
N
SO2
N
B
A
Cl
Cl
R = phenyl, cyclohexyl, 1-naphthyl, benzyl, benzoyl
X = O or S
Y = C=O or phenyl
H
Cl
N
X
H
N
z
Y
SO2
C
Sulofenur: X = O, Y = H, Z = CH2
D
E
LY295501: X = O, Y = Cl, Z = O
X = S, Y = H, Z = CH2
Fig. 1 Structures of some studied substituted benzenesulfonamide and sulfonylureas

302
Med Chem Res (2007) 16:300–318
functionalities were maintained as basic structure features. The 4-chlorophenyl
moiety at position 3 of the indenopyrazoline part (A; Fig. 1) was replaced by either
thiophene or pyridine rings, which are well acknowledged for their chemother-
apeutic effects (Zhao et al., 2006; Rimagnoli et al., 2006; Singh et al., 2007;
Srivastava et al., 2007), which would enhance the anticipated bioactivity.
Cyclization of some thioureido derivatives into the corresponding 5-membered
thiazolidinons and thiazolines, which are famous for their antitumor activity (B;
Fig. 1) (El-Sabbagh et al., 1999; Yalowitz et al., 2002; Mahboobi et al., 2006;
Cejudo et al., 2006), resulted in some moderately active antitumor analogs
(Rostom, 2006). In the present investigation, it was considered of interest to
incorporate some of the new thioureido derivatives into the the six-membered
thiazinone in addition to the five-membered thiazolidinone and thiazoline ring
systems, in order to explore the influence of such structure modification on the
anticipated biological activity. All these efforts are attempted with the anticipation
of discovering new structure leads that might be in use in the design of new,
potent, and selective anticancer agent(s).
Chemistry
The synthetic strategies utilized for the synthesis of the intermediate and target
products are depicted in Schemes 1 and 2. In Scheme 1, the starting chalcones
1 and 2 were obtained by condensing equimolar amounts of 1-indanone with
2-thiophencarboxaldehyde or 3-pyridinecarboxaldehyde, respectively, in an aqueous
ethanolic solution of sodium hydroxide according to a previously described
procedure (Makki and Faidallah, 1993). Cyclocondensation of these chalcones with
4-hydrazinobenzenesulfonamide hydrochloride in boiling ethanol afforded the
indeno[1,2-c]-pyrazolines
3 and 4 in good yields. The targeted indeno
[1,2-c]pyrazoles 5 and 6 could be achieved by mild oxidation of the substituted
indeno[1,2-c]pyrazolines 3 and 4 with bromine water at room temperature. The
substituted sulfonylureas 7–12 were successfully obtained by condensing the key
intermediate sulfonamides 3 and 4 with the appropriate isocyanate in the presence of
anhydrous potassium carbonate as a mild basic catalyst. Furthermore, the targeted
substituted 3-(2-thienyl)-indeno[1,2-c]pyrazole benzenesulfonylureas 13–15 could
be synthesized via oxidation of the substituted indeno[1,2-c]pyrazoline benze-
nesulfonylureas 7–9 (R = 2-thienyl) with bromine water at room temperature.
Moreover, condensing the key intermediates 3 and 4 with different isothiocy-
anates in the presence of anhydrous potassium carbonate resulted in the formation of
the substituted benzenesulfonylthioureas 16–21. At this stage, cyclization of the
sulfonylthioueido derivatives 16–19 (Scheme 2) with ethyl bromoacetate in the
presence of anhydrous sodium acetate afforded the corresponding thiazolidin-4-ones
22–25. Furthermore, reacting 16–19 with phenacyl bromide under similar reaction
conditions resulted in the formation of the desired 1,3-thiazolines 26–29. In an
analogous fashion, reacting the sulfonylthioueido intermediates 16–19 with ethyl 3-
bromopropionate or 1,3-dichloroacetone in the presence of anhydrous sodium

Med Chem Res (2007) 16:300–318
303
H
H
N
O
R1
NH2
SO2
N
N
N
N
SO2
N
R
S
5: R = 2-thienyl; 6: R = 3-pyridyl
13: R1 = cyclohexyl; 14: R1 = phenyl
15: R1 = 1-naphthyl
Br2 / H2O
Br2 / H2O
R = 2-Thienyl
NHNH2
H
H
N
O
R1
.
HCl
O
N
NH2
N
N
SO2NH2
R1NCO
SO2
N
SO2
N
R
R
R
7: R = 2-thienyl, R1 = cyclohexyl;
8: R = 2-thienyl, R1 = phenyl;
9: R = 2-thienyl, R1 = 1-naphthyl;
3: R = 2-thienyl; 4: R = 3-pyridyl
1: R = 2-thienyl; 2: R = 3-pyridyl
10: R = 3-pyridyl, R1 = cyclohexyl;
11: R = 3-pyridyl, R1 = phenyl;
12: R = 3-pyridyl, R1 = 1-naphthyl
H
H
N
S
R2
O
N
R2NCS
N
SO2
+
R-CHO
R = 2-Thienyl or 3-pyridyl
N
R
16: R = 2-thienyl, R2 = phenyl;
17: R = 2-thienyl, R2 = benzyl;
18: R = 2-thienyl, R2 = benzoyl;
19: R = 3-pyridyl, R2 = phenyl;
20: R = 3-pyridyl, R2 = benzyl;
21: R = 3-pyridyl, R2 = benzoyl
Scheme 1 Synthesis of compounds 1–21
acetate furnished the targeted [1, 3]thiazinan-4-ones 30–33 and the [1, 3]thiazinan-
5-ones 34–37, respectively.
Results and Discussion
Primary in Vitro One-Dose Three-cell-line Assay
Primary in vitro one-dose anticancer assay was performed using the three-cell-line
panel consisting of NCI-H460 (lung), MCF7 (breast), and SF-268 (CNS) in
accordance with the protocol of the Drug Evaluation Branch, NCI, Bethesda
(Grever et al 1992; Boyd and Paul 1995; Monks et al 1991). Out of the newly
synthesized compounds, 28 compounds, namely 3–8, 10, 11, 13, 14, 16–21, 22, 24–
26, 28–30, 32–34, 36, and 37, were selected by the National Cancer Institute (NCI)
in vitro disease-oriented human cells screening panel assay to be evaluated for their
in vitro antitumor activity. All the compounds which reduced the growth of any one
of the cell lines to 32% or less were passed on for evaluation in the full panel of 60
human tumor cell lines. The results revealed that 13 compounds, namely 3, 5, 11,
13, 14, 16, 18, 24, 26, 28, 30, 32, and 34, passed this primary anticancer assay and
consequently were carried over to be tested against a panel of 56 different tumor cell
lines.

304
Med Chem Res (2007) 16:300–318
R2
H
R2
O
N
H
N
S
R2
N
N
N
N
N
N
N
S
N
SO2
N
SO2
S
N
SO2
R
R
O
R
O
Br
Br
O
22: R = 2-thienyl, R2 = phenyl;
23: R = 2-thienyl, R2 = benzyl;
24: R = 2-thienyl, R2 = benzoyl;
25: R = 3-pyridyl, R2 = phenyl
16: R = 2-thienyl, R2 = phenyl;
17: R = 2-thienyl, R2 = benzyl;
18: R = 2-thienyl, R2 = benzoyl;
19: R = 3-pyridyl, R2 = phenyl
26: R = 2-thienyl, R2 = phenyl;
27: R = 2-thienyl, R2 = benzyl;
28: R = 2-thienyl, R2 = benzoyl;
29: R = 3-pyridyl, R2 = phenyl
R2
O
N
S
R2
N
N
N
N
SO2
N
O
N
N
SO2
S
R
R
O
O
Cl
Cl
Br
O
34: R = 2-thienyl, R2 = phenyl;
35: R = 2-thienyl, R2 = benzyl;
30: R = 2-thienyl, R2 = phenyl;
31: R = 2-thienyl, R2 = benzyl;
32: R = 2-thienyl, R2 = benzoyl;
33: R = 3-pyridyl, R2 = phenyl
36: R = 2-thienyl, R2 = benzoyl;
37: R = 3-pyridyl, R2 = phenyl
Scheme 2 Synthesis of the target compounds 22–37
In Vitro Full-panel 60-cell-line Assay
Fifty six cell lines of nine tumor subpanels, including leukemia, non-small-cell lung,
colon, CNS, melanoma, ovarian, renal, prostate, and breast cancer cell lines, were
incubated with five concentrations (0.01–100 lM) for each compound and used to
create log concentration–percentage growth inhibition curves. Three response
parameters (GI50, TGI, and LC50) were calculated for each cell line. The GI50 value
(growth inhibitory activity) corresponds to the concentration of the compounds
causing 50% decrease in net cell growth, the TGI value (cytostatic activity) is the
concentration of the compounds 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 hours). Subpanel and full-
panel mean-graph midpoint values (MG-MID) for certain agents are the average of
individual real and default GI50, TGI, or LC50 values of all cell lines in the subpanel
or the full panel, respectively.
In the present study, the active compounds exhibited remarkable antitumor
activities against most of the tested subpanel tumor cell lines (GI50 , TGI, and LC50
values less than 100 lM). These compounds showed a distinctive pattern of
sensitivity against some individual cell lines (Table 1), as well as a broad spectrum
of antitumor activity (Tables 2 and 3).
Concerning the sensitivity against some individual cell lines (Table 1),
compound 3 showed high activity against non-small-cell lung NCI-H322 cell line
(GI50 5.79 lM). The analog 5 displayed a distinguished sensitivity profile towards
leukemia cell lines with a GI50 range of 0.86–6.68 lM, in addition to a remarkably
high activity against the non-small-cell lung cancer NCI-H226 cell line with a GI50
value of 4.90 lM. Compound 13 exhibited a significant activity against the
leukemia CCRF-CEM cell line with a GI50 value of 1.15 lM. Moreover, derivative
14 proved to be sensitive against the non-small-cell lung NCI-H322 and leukemia
RPMI-8226 cell lines (GI50 values of 4.50 and 5.49 lM, respectively). Whereas,

Med Chem Res (2007) 16:300–318
305
a
Table 1 Growth inhibitory concentration (GI50, lM) in the 60 tumor cell lines assay
Cell lines
3
5
11
13
14
16
18
24
26
28
30
32
34
Leukemia
CCRF-CEM
K-562
31.8 4.24 40.9 1.15 13.6 30.5 27.9 37.5 50.3 41.3 28.0 15.0 36.8
b
c
57.5 0.86 26.6 10.0 14.0 31.3 NT
NT
—
27.6
—
26.4 10.3
MOLT-4
RPMI-8226
18.9 1.43 11.6 49.9 15.5 17.5 26.2 22.2 19.0 30.5 47.0 50.4 0.01
32.3 6.68 27.1 10.7 5.49 30.3 29.7 34.0 51.0 32.3 18.1 58.6 31.0
Non-small-cell lung cancer
A549/ATCC
EKVX
13.1 20.7 29.2 28.0 20.8 15.7 20.3 70.5 57.0 39.5 14.4 10.6 47.2
15.9 11.9 27.8 28.6 26.1 19.5 19.6 42.4 35.5 40.7 25.7 22.1 39.5
16.9 16.5 20.4 18.1 20.3 NT 32.9 73.2 23.7 18.6 31.1 17.4
25.8 5.56 74.9 14.3 12.3 24.9 53.0 37.3 35.9 25.4 79.9
HOP-62
—
HOP-92
—
—
NCI-H226
NCI-H23
NCI-H322
NCI-H460
NCI-H522
Colon cancer
COLO 205
HCC-2998
HCT-116
HCT-15
NT 4.90 23.7 21.3 19.2 27.2 NT
20.3 11.9 20.8 12.8 12.2 17.6 NT
58.5 78.9 30.1 26.5 15.9 22.3
25.1 68.0 13.1 16.7 17.2 24.2
5.79 11.9 17.3 37.9 4.50 56.6 14.2 16.6 31.4 22.3 12.8 32.5 16.9
38.5 93.0 18.9 24.6 95.1 26.6
39.3 12.4 17.7 12.3 31.5 32.8 20.5 60.1 61.9 23.7 15.9 14.4 23.8
—
—
—
—
—
—
—
58.9 25.3 31.1 23.3 44.0 41.9 38.4 51.0
—
98.1 36.8 66.5 57.2
14.8 12.8 17.3 6.63 15.3 13.9 NT
10.7 7.67 15.3 31.6 9.13 15.1 NT
17.6 32.8 14.0 17.9 18.2 20.5
22.0 NT 30.5 20.3 10.0 12.5
34.7 12.0 23.1 17.8 16.9 22.6 30.2 56.3 67.4 61.4 15.9 15.1 35.5
26.1 22.9 37.9 34.3 26.7 36.2 46.0 76.0 82.0 73.3 37.0 39.6 61.0
15.7 13.5 24.8 34.9 8.08 21.5 28.2 21.4 32.1 38.3 17.1 29.4 28.4
35.4 10.7 34.0 20.7 22.9 30.0 34.8 38.5 94.2 58.8 24.0 13.1 45.2
HT29
KM12
SW-620
CNS cancer
SF-268
—
—
62.1
—
26.0 30.7
—
—
—
—
—
13.7 —
SF-295
16.2 13.8 20.6 34.9 21.9 18.9 25.4 31.7 54.5 37.1 15.8 32.7 32.7
SF-539
17.4 21.1 30.6 27.6 25.0 22.7 27.9 44.6
34.4 18.1 83.9 56.8 33.8 46.3 77.0
31.1 25.5 52.2 70.4 71.9
—
—
—
58.6 21.8 23.8 27.8
59.2 22.3 45.7 55.7
SNB-19
—
SNB-75
—
—
—
—
30.2
—
—
U251
19.4 12.3 31.9 25.6 22.9 26.2 20.1 36.3 31.2 32.0 14.4 4.50 28.0
Melanoma
LOX IMVI
M14
19.1 6.58 15.4 5.66 12.0 24.4 15.2 22.7 32.4 30.7 13.6 7.61 21.2
26.0 17.2 24.9 64.1 25.0 22.7 21.2 44.7 81.5 33.0 15.3 10.8 39.1
SK-MEL-2
SK-MEL-28
SK-MEL-5
UACC-257
UACC-62
Ovarian cancer
IGROV1
OVCAR-3
—
27.6 35.6 23.6 70.1 55.9 44.0
—
—
—
—
—
—
28.0 29.4 60.2
42.1 50.2
75.6 25.8 71.5 34.2 50.2 53.8 84.0
—
18.0 10.5 23.6 18.7 15.6 20.8 16.9 22.7 48.0 24.4 14.1 2.56 38.2
29.7 23.2 26.6 24.8 17.7 29.1 42.8 45.7 24.2 0.33 42.3
21.8 11.5 17.8 19.3 15.8 18.5 14.6 23.2 60.3 27.3 16.7 6.37 16.1
—
—
30.0 2.49 16.2 4.12 21.4 65.3 3.14 28.3 NT
24.8 14.4 21.6 24.5 21.8 17.2 30.6 59.2
—
17.4 14.5 19.5
—
55.2 25.9 27.4
—

306
Med Chem Res (2007) 16:300–318
Table 1 continued
Cell lines
3
5
11
—
13
14
16
18
24
—
26
—
28
—
30
32
—
34
—
OVCAR-4
OVCAR-5
OVCAR-8
SK-OV-3
Renal cancer
786-0
—
29.9
27.0 34.9 69.8 87.6
49.3
26.6 18.5 26.8 26.6 20.1 17.9 18.7 32.2 88.9 34.3 22.4 25.8 34.5
15.3 20.5 27.0 25.3 28.3 39.0 20.3 45.7 47.2 42.9 19.2 0.81 25.4
48.0 37.7
—
44.5 60.8 49.5 90.8
—
—
—
32.9 52.8 —
17.6 17.1 35.5 32.3 17.6 19.7 27.9 48.8 64.2 21.8 64.2 14.1 22.1
A 498
NT NT NT NT NT NT NT
19.8 11.7 23.6 17.3 17.6 26.4
NT NT NT NT NT NT
23.6 42.5 43.8 42.5 5.18 22.1
ACHN
—
CAKI-1
19.6 10.1 26.4 16.0 11.1 18.5 26.1 39.0 73.0 38.0 73.0 26.8 74.5
20.6 17.4 45.3 29.2 85.3 78.1 54.0 63.9 98.2 37.0 98.2 6.31 41.0
23.5 12.2 30.4 21.5 19.9 21.4 26.0 90.7 94.3 44.4 94.3 16.7 34.3
RXF 393
SN12C
TK-10
13.2 12.2 11.7 17.5 4.99 12.2 NT
39.4 12.2 48.8 15.4 32.2 51.2 41.1
49.6 32.7 30.5 32.7 24.5 NT
98.1 30.7 69.3
UO-31
—
—
—
Prostate cancer
PC-3
22.1 20.6 36.2 14.5 10.4 22.9 29.7 38.1 75.0 39.2 75.0 29.2 45.0
DU-145
14.5 24.1 36.6 31.9 28.6 17.8 28.3 38.2
—
47.0
—
23.2 30.7
Breast cancer
MCF7
—
—
27.0
—
13.4 3.10
—
—
—
29.1
—
—
—
29.1 9.10 88.8
32.9
NCI/ADR-RES
17.1 12.4 64.4 20.4 16.1 19.3 18.8
—
—
MDA-MB-231/ATCC 16.8 17.3 19.8 27.9 15.3 24.8 22.5 44.6 52.1 41.3 52.1 5.77 21.4
HS 578T
MDA-MB-435
BT-549
T-47D
28.7 16.1 40.7 37.5 16.8 20.9 16.1 25.3 60.6 17.0 60.6 2.85 20.5
19.3 11.8 21.4 32.5 22.1 19.4 21.7 37.4 38.7 31.1 38.7 22.7 29.5
19.3 12.4 30.6 33.7 24.1 24.6 21.6 33.5 47.5 69.7 47.5 4.77 21.4
66.9 12.8 93.0 39.7 37.9
—
69.6
—
—
—
—
—
69.2
a
Data obtained from NCI’s in vitro disease-oriented human tumor cell screen
NT: not tested
b
c
GI50 value greater than 100 lM
compound 16 showed good activity against the breast MCF7 cell line (GI50 value of
3.10 lM). Compound 32 exhibited potential activity against melanoma UACC-257
and ovarian OVCAR-8 cell lines, with GI50 values of 0.33 and 0.81 lM,
respectively. Furthermore, compound 34 showed the highest growth inhibitory
activity against the leukemia MOLT-4 cell line with a GI50 value of *0.01 lM. On
the other hand, the ovarian IGROV1 cell line proved to be very sensitive to
compounds 5, 13, and 18, with GI50 values of 2.49, 4.12, and 3.14 lM, respectively.
With regard to broad spectrum antitumor activity, the results revealed that all of
therteen active compounds, i.e., 3, 5, 11, 13, 14, 16, 18, 24, 26, 28, 30, 32 and 34;
revealed effective growth inhibition GI50 (MG-MID) values of 25.7, 13.2, 30.9,
22.9, 20.4, 25.7, 30.9, 46.8, 54.9, 42.6, 24.5, 15.1 and 33.1 lM, respectively
(Table 2), beside a cytostatic activity TGI (MG-MID) values of 63.1, 33.1, 77.6,
60.2, 67.6, 66.1, 79.4, 89.1, 97.7, 91.2, 54.9, 56.2, and 87.1 lM, respectively
(Table 3).

Med Chem Res (2007) 16:300–318
307
Table 2 Median growth inhibitory concentrations (GI50, lM) of in vitro subpanel tumor cell linesa
Cpd. No.
Subpanel tumor cell linesb
MG-MIDc
I
II
III
IV
V
VI
VII
VIII
IX
3
35.1
29.6
21.7
30.0
38.7
18.2
30.5
37.9
49.3
66.8
43.7
29.6
21.7
41.2
28.0
14.9
26.2
24.2
20.4
25.9
35.5
40.4
68.1
53.3
24.1
27.4
37.2
36.4
31.8
54.8
49.5
44.4
34.0
53.3
64.9
80.9
64.5
34.1
36.7
57.4
41.5
17.5
30.8
27.2
29.5
32.2
34.1
73.3
74.6
51.6
22.0
15.3
45.3
40.8
20.6
48.6
25.3
31.2
43.0
41.9
60.9
87.2
72.0
27.8
36.9
63.2
21.9
13.3
31.7
21.3
29.6
32.5
45.8
59.4
72.1
44.8
72.1
17.7
43.9
18.3
22.3
36.4
23.2
19.5
20.3
29.0
38.1
87.5
43.1
87.5
26.2
37.8
38.3
26.1
42.4
41.7
20.8
30.8
38.6
63.0
61.1
65.3
61.1
25.4
50.1
25.7
13.2
30.9
22.9
20.4
25.7
30.9
46.8
54.9
42.6
24.5
15.1
33.1
5
3.30
11
13
14
16
18
24
26
28
30
32
34
a
26.5
17.9
12.1
27.4
27.9
31.2
55.0
32.9
48.3
37.6
19.5
Data obtained from NCI’s in vitro disease-oriented human tumor cell screen
b
I, Leukemia; II, non-small-cell lung cancer; III, colon cancer; IV, CNS cancer; V, melanoma; VI,
ovarian cancer; VII, renal cancer; VIII, prostate cancer; IX, breast cancer
c
GI50 (lM) full panel mean-graph mid point (MG-MID) = the average sensitivity of all cell lines towards
the test agent
In addition, except for compounds 24, 26, 28 and 34, the rest of the active
compounds; 3, 5, 11, 13, 14, 16, 18, 30 and 32 displayed a weak cytotoxic pattern
with LC50 (MG-MID) values of 95.5, 69.2, 97.7, 95.4, 95.5, 97.7, 97.7, 87.1, and
89.1 lM, respectively (Tables 2 and 3).
Further interpretation of the data presented in Tables 2 and 3 revealed that,
compound 5 [GI50, TGI, and LC50 (MG-MID) values of 13.2, 33.1, and 69.2 lM,
respectively] proved to be the most active member in this study with potential
activity against all the tested subpanel tumor cell lines and particular effectiveness
on the leukemia subpanel at both the GI50 (3.30 lM), and the TGI levels (15.7 lM).
Compound 13 displayed a reasonable growth inhibition and cytostatic effects,
however, its showed weak cytotoxic activity [GI50, TGI and LC50 (MG-MID) values
of 22.9, 60.2, and 95.4 lM, respectively]. It proved to be sensetive toward the
leukemia subpanel as evidenced by its GI50 and TGI (MG-MID) values (17.9 and
26.6, respectively). Furthermore, compounds 3 [GI50, TGI, and LC50 (MG-MID)
values of 25.7, 63.1, and 95.5 lM, respectively) and 16 (GI50, TGI, and LC50 (MG-
MID) values of 25.7, 66.1, and 97.7 lM, respectively) were nearly equipotent, with
almost the same range of activity against most of the tested subpanel tumor cell
lines. On the other hand, compound 26 was found to be the least effective antitumor
agent in the present investigation with GI50 and TGI (MG-MID) values of 54.9 and
97.7 lM, respectively, and without any cytotoxic effect.
The ratio obtained by dividing the full panel MG-MID (lM) of the compounds
by their individual subpanel MG-MID (lM) is considered as a measure of

308
Med Chem Res (2007) 16:300–318
Table 3 Median total growth inhibitory concentrations (TGI, lM) of in vitro subpanel tumor cell linesa
Cpd. no.
Subpanel tumor cell linesb
MG-MIDc
I
II
III
IV
V
VI
VII
VIII
IX
3
95.5
15.7
91.4
26.6
85.6
85.0
77.2
84.5
90.9
80.3
93.9
80.1
82.1
45.5
31.3
56.4
46.7
74.4
48.4
78.9
85.9
98.2
83.9
47.4
71.4
79.0
74.5
37.2
78.7
73.3
37.2
75.9
85.6
79.8
96.9
90.4
60.7
52.6
66.3
70.3
42.0
87.6
93.6
80.7
77.5
90.9
—
51.0
38.9
49.3
53.4
43.0
68.4
67.7
—
79.6
37.3
90.8
80.3
98.3
81.3
95.3
—
43.7
29.7
86.4
40.1
42.5
47.0
87.2
99.1
96.5
95.3
43.2
38.1
96.4
48.6
22.3
52.1
34.7
87.8
94.2
56.5
57.3
83.7
—
63.1 (95.5)d
33.1 (69.2)
77.6 (97.7)
60.2 (95.4)
67.6 (95.5)
66.1 (97.7)
79.4 (97.7)
89.1
5
e
11
13
14
16
18
24
26
28
30
32
34
a
—
65.9
98.1
47.9
—
—
—
—
—
—
—
97.7
—
98.2
33.8
21.2
84.3
—
—
—
91.2
54.1
68.2
99.7
55.4
42.1
-
53.9
—
56.2
73.5
95.5
54.9 (87.1)
56.2 (89.1)
87.1
—
Data obtained from NCI’s in vitro disease-oriented human tumor cell screen
b
c
For subpanel tumor cell lines, see footnote (b) of Table 2
TGI (lM) full panel mean-graph mid point (MG-MID) = the average sensitivity of all cell lines towards
the test agent
d
LC50 (lM) full panel mean-graph mid point (MG-MID) = the average sensitivity of all cell lines
towards the test agent
e
TGI (MG-MID) value greater than 100 lM.
compound selectivity. Ratios of 3–6 refer to moderate selectivity, ratios greater than
6 indicate high selectivity towards the corresponding cell line, while compounds not
meeting either of these criteria are rated as nonselective (Acton et al., 1994).
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 0.40–2.60 and 0.66–2.26 at the
GI50 and TGI levels, respectively, except compound 5, which showed moderate
selectivity towards the leukemia subpanel at both the GI50 and TGI levels
(selectivity ratios of 4.0 and 2.10, respectively). However, compound 13 revealed
mild selectivity towards the leukemia subpanel with a selectivity ratio near 3 (2.60
and 2.26, at the GI50 and TGI leves, respectively). A close study of the structures of
the active compounds revealed that the indeno[1,2-c]pyrazole moiety contains two
different heteocylic counterparts: the thiophen and the pyridine ring systems. From
the NCI data (Tables 1–3), it appears that the thienyl moiety is the most favorable
substituent (12 active compounds) rather than the pyridyl one (compound 11). The
key intermediate 4-(3a,4-dihydro-3-(2-thienyl)-3H-indeno[1,2-c]pyrazol-2-yl)-ben-
zene-sulfonamide 3 showed considerable antitumor activity as indicated from its
GI50, TGI, and LC50 (MG-MID) values (25.7, 63.1 and 95.5 lM, respectively).
Oxidation of 3 gave rise to the corresponding 4-(3-(2-thienyl)-3H-indeno[1,2-
c]pyrazol-2-yl)-benzenesulfonamide 5, which proved to be the most active member

Med Chem Res (2007) 16:300–318
309
in this series with the highest growth inhibitory, cytostatic and cytotxic potentials
(GI50, TGI and LC50 (MG-MID) values of 13.2, 33.1, and 69.2, respectively;
Tables 2 and 3), with special effectiveness on the leukemia subpanel at the GI50 and
TGI (MG-MID) levels (3.30 and 15.7 lM, respectively). These findings were
concordant with the fact that the sulfonamido functionality is associated with
antitumor activity (Supuran et al., 2001; Hu et al., 2006; Brzozowski et al., 2007).
Condensing the parent compound 3 with different isocyanates resulted in the
formation of the disubstituted sulfonylureido derivatives 7–12, of which only one
analog (11; R = 3-pyridyl; R1 = phenyl) exibited reasonable antitumor activity
[GI50, TGI and LC50 (MG-MID) values of 30.9, 77.6 and 97.7, respectively;
Tables 2 and 3], whereas the other congeners 7, 8, and 10 proved to be totally
inactive. The synthesis of the oxidized pyrazole derivatives 13–15 gave rise to two
remarkably active N1-substituted sulfonylureido-pyrazole derivatives 13 and 14.
Such derivatization led to about a twofold reduction in the overall antitumor activity
when they are compared with the parent pyrazole 5. The type of substituent in this
type of sulfonylureas did not noticeably affect the antitumor activity as observed
from their GI50, TGI, and LC50 values, which lie nearly in the same range (Tables 2
and 3). However, compound 13 (R = 2-thienyl; R1 = cyclohexyl) proved to be more
senstive than 14 (R = 2-thienyl; R1 = phenyl) toward the leukemia subpanel at the
GI50 and TGI (MG-MID) levels (8.9 versus 12.1 and 26.6 versus 85.6 lM,
respectively; Tables 2 and 3). Conversion of the sulfonamide 3 to the N1-substituted
sulfonylthioureido derivatives 16–21 furnished two active compounds (16; R = 2-
thienyl, R2 = phenyl) and (18; R = 2-thienyl, R2 = benzoyl). Both compounds
displayed almost the same antitumor profile at the GI50 and LC50 (MG-MID) levels
(25.7 versus 30.9 and 97.7 lM, respectively). Meanwhile, compound 16 proved to
exert slightly better cytostatic activity then its analog 18 [TGI (MG-MID) values
66.1 versus 79.4 lM, respectively]. Here it should be pointed out that, introducing a
thioureido functionality in 16 and 18 did not result in a noticable effect on the
overall antitumor activity when compared with the parent sulfonamide 3 (Tables 2
and 3). On the other hand, the thioureido group in compounds 16–19 was partly
incorporated in a rigid structure, either in the form of five-membered 2-oxothiaz-
olidine 22–25, 1,3-thiazoline 26–29 or the six-membered [1, 3]thiazinan-4-one 30–
33, [1, 3]thiazinan-5-one 34–37 ring systems. The results clearly showed that the
six-membered [1, 3]thiazinanones displayed much better antitumor activity than the
five-membered thiazole derivatives. In this view, within the [1, 3]thiazinan-4-one
30–33 series, compounds 30 [GI50, TGI, and LC50 (MG-MID) values of 24.5, 54.9,
and 87.1 lM, respectively] and 32 [GI50, TGI, and LC50 (MG-MID) values of 15.1,
56.2, and 89.1 lM, respectively] revealed good antitumor spectrum with nearly
equal cytostatic and cytotoxic effects and better growth inhibitory activity for
compound 32 (R = 2-thienyl; R2 = benzoyl). In addition, compound 32 showed
about a twofold improvement in the growth inhibitory as well as the cytostatic
activity when compared with the parent sulfonylthioureido derivative 18, as
evidenced by their GI50 and TGI (MG-MID) values (15.1 versus 30.9 and 56.2
versus 79.4 lM, respectively), wheras its cytotoxic effecacy was slightly improved
[LC50 (MG-MID) values of 89.1 and 97.7 lM, respectively]. Meanwhile, the
[1, 3]thiazinan-5-one analog (34, R = 2-thienyl; R2 = phenyl) proved to be weaker

310
Med Chem Res (2007) 16:300–318
than the six-membered analogs 30 and 32, with growth inhibitory and cytostatic
activity [GI50 and TGI (MG-MID) values of 33.1 and 87.1 lM, respectively] and
without any cytotoxic effect. Furthermore, incorporation of the thioureido
counterpart in compounds 16–19 into the five-membered 2-oxothiazolidines and
1,3-thiazolines, led to three active compounds, namely 24, 26, and 28, with a
remarkable decrease in the overall growth inhibitory and cytostatic activity profiles
and complete abolishment of the cytotoxic activity. Among these, compound 26
could be considered as the least active member in this investigation [GI50 and TGI
(MG-MID) values of 54.9 and 97.7 lM, respectively].
Conclusion
The main aim of the present study was to synthesize and investigate the antitumor
activity of new compounds incorporating the sulfonamido, N1, N3-disubstituted
sulfonylurea and thiourea functionalities structurally related to a previously studied
series of compounds A and B and the well-documented sulfonylurea anticancer
agents Sulofenur C and its structure analogs D and E (Fig. 1). This aim could be
achieved by the synthesis of hybrid compounds comprising the above-mentioned
pharmacophores substituted essentially with a 3-(2-thienyl or 3-pyridyl)-indeno
[1,2-c]pyrazol(in)e counterpart at the N3 aryl moiety for synergistic purpose. The
results revealed that thirteen compounds, i.e., 3, 5, 11, 13, 14, 16, 18, 24, 26, 28, 30,
32 and 34, showed promising broad-spectrum antitumor activity. In general,
compounds comprising the thienyl moiety displayed better antitumor spectra than
the pyridyl one (only one active compound; 11). The oxidized 3-(2-thienyl)-
indeno[1,2-c]pyrazole benzenesulfonamide and N1,N3-disubstituted sulfonylurea
derivatives appeared to exhibit better antitumor activity than their pyrazoline
analogs. Bioisosteric replacement of the sulfonylureido functionality with a
sulfonylthioureido did not significantly influence the antitumor activity. Incorpo-
ration of the sulfonylthioureido pharmacophore within five-membered ring systems
(compounds 24, 26, and 28) led to an obvious reduction in the antitumor activity.
Meanwhile, the inclusion into a 6-membered ring either keeps (compounds 30 and
34) or even improves (compound 32) the activity. Except for compounds 24, 26, 28,
and 34, the remaining nine active compounds showed some mild cytotoxic efficacy
(Table 3). Compound 5, 4-(3-(2-thienyl)-3H-indeno[1,2-c]pyrazol-2-yl)-benzene-
sulfonamide, proved to be the most active member in this study with potential
activity against all the tested subpanel tumor cell lines and particular effectiveness
on the leukemia and prostate subpanels at both the GI50 and the TGI levels.
Experimental
Melting points were determined in open glass capillaries on a Gallenkamp melting-
point apparatus and were uncorrected. The infrared (IR) spectra were recorded on a
Perkin-Elmer 297 infrared spectrophotometer using the KBr plate technique. The
1H-NMR spectra were recorded on a Varian EM 360 spectrometer using

Med Chem Res (2007) 16:300–318
311
tetramethylsilane as an internal standard and DMSO-d6 as the solvent (Chemical
shifts in d, ppm). Splitting patterns were designated as follows: s: singlet; d: doublet;
m: multiplet. Elemental analyses were performed at the Microanalytical Unit,
Faculty of Science, King Abdul-Aziz University, Jeddah, Saudi Arabia, and the
values found were within
0.4% of the theoretical values. Follow up of the
reactions and checking the homogeneity of the compounds were made by TLC on
silica gel-protected aluminum sheets (Type 60 F254, Merck) and the spots were
detected by exposure to an ultraviolet (UV) lamp at a wavelength of 254 nm.
4-(3a,4-Dihydro-3-substituted-3H-indeno[1,2-c]pyrazol-2-yl)-
benzenesulfonamides (3, 4).
A solution of the appropriate chalcone 1 or 2 (0.02 mol) in ethanol was refluxed
with 4-hydrazino-benzenesulfonamide hydrochloride (4.9 g, 0.022 mol) for 6 h. The
volume of the reaction mixture was concentrated down to half the volume and the
separated product was filtered, washed with cold ethanol, and recrystallized from
benzene-ethanol (1:1). Physicochemical and analytical data are recorded in Table 4.
IR (cm–1): 3200 and 3360 (NH2), 1185 and 1340 (SO2N). 1H-NMR (d, ppm) for 3:
2.28 (d, J = 9 Hz, 2H, C4-2H), 2.52 (m, 1H, C3a-H), 5.52 (m, 1H, C3-H), 7.14–8.70
(m, 11H, Ar-H), 10.54 (s, 2H, NH2); for 4: 2.24 (d, J = 9 Hz, 2H, C4-2H), 2.64 (m,
1H, C3a-H), 5.55 (m, 1H, C3-H), 6.58–8.66 (m, 12H, Ar-H).
4-(3-Substituted-4H-indeno[1,2-c]pyrazol-2-yl)-benzenesulfonamides (5, 6)
To a stirred suspension of the pyrazoline 3 or 4 (0.01 mol) in water (10 mL), bromine
water (5%, 15 mL) was gradually added over a period of 30 min at room temperature.
After stirring for an additional 5h, the pyrazole derivative was formed, collected by
filtration, washed with water, and dried. It was recrystallized from ethanol.
Physicochemical and analytical data are recorded in Table 4. IR (cm–1): 3230 and
3345 (NH2), 1315 and 1190 (SO2N). 1H-NMR (d, ppm) for 5: 4.03 (s, 2H, C4-2H),
7.56-8.23 (m, 11H, Ar-H); for 6: 3.89 (s, 2H, C4-2H), 7.11–8.86 (m, 12H, Ar-H).
N1-Substituted N3-[4-(3-substituted-3a,4-dihydro-3H-indeno[1,2-c]pyrazol-2-
yl)-benzenesulf-onyl]-ureas (7–12).
A mixture of the pyrazoline 3 or 4 (0.005 mol) and anhydrous potassium carbonate
(1.4 g, 0.01 mol) in dry acetone (20 mL) was heated under reflux with stirring with
the appropriate isocyanate (0.0055 mol) for 18 h. The solvent was removed under
reduced pressure and the remaining solid residue was dissolved in water (30 mL).
After acidification of the resulting solution with 2N hydrochloric acid, the
precipitated crude product was filtered, washed with water, dried and recrystallized
from ethanol. Physicochemical and analytical data are recorded in Table 4. IR (cm–
1): 1300–1330 and 1165–1180 (SO2N), 1640–1670 (C=O). 1H-NMR (d, ppm) for 7:
1.35–2.55 (m, 11H, cyclohexyl), 2.22 (d, J = 9 Hz, 2H, C4-2H), 2.61 (m, 1H, C3a-

312
Med Chem Res (2007) 16:300–318
Table 4 Physicochemical and analytical data of compounds 3–37
Cpd,
R
R1 or R2
Y. (%) M.p.
(°C)
Mol. formula
(mol. weight)
Analysis
(calculated/found)
3
2-Thienyl
3-Pyridyl
2-Thienyl
3-Pyridyl
—
—
—
—
80
78
84
73
186-7
310-1
168-9
266-7
138-9
130-2
C
20H17N3O2S2 60.74 4.33 10.62 16.22
(395.50) 60.53 4.47 10.40 16.35
21H18N4O2S
(390.46)
4
C
64.60 4.65 14.35
64.43 4.81 14.44
8.21
8.05
5
C
20H15N3O2S2 61.05 3.84 10.68 16.30
(393.48) 61.17 3.63 10.90 16.07
21H16N4O2S
(388.44)
6
C
64.93 4.15 14.42
65.11 4.08 14.70
8.25
8.06
7
2-Thienyl Cyclohexyl 70
C
27H28N4O3S2 62.28 5.42 10.76 12.32
(520.67) 62.04 5.60 10.55 12.48
27H22N4O3S2 63.02 4.31 10.89 12.46
8
2-Thienyl Phenyl
75
C
(514.62)
63.15 4.22 11.02 12.33
9
2-Thienyl 1-Naphthyl 78
142-3 C31H24N4O3S2 65.94 4.28
(564.68)
28H29N5O3S
(515.63)
28H23N5O3S
(509.58)
32H25N5O3S
(599.64)
9.92 11.36
11.14
66.03 4.15 10.9
10
11
12
13
14
15
16
17
18
19
20
21
22
3-Pyridyl
3-Pyridyl
3-Pyridyl
Cyclohexyl 65
304-5
164-6
177-8
132-3
125-7
164-6
134-5
128-9
124-6
C
65.22 5.67 13.58
6.22
6.40
6.29
6.37
5.73
5.68
65.38 5.51 13.33
66.00 4.55 13.74
66.08 4.49 13.55
68.68 4.50 12.51
68.40 4.77 12.60
Phenyl
69
C
1-Naphthyl 72
C
2-Thienyl Cyclohexyl 70
C
27H26N4O3S2 62.53 5.05 10.80 12.37
(518.65) 65.77 5.01 10.65 12.43
27H20N4O3S2 63.26 3.93 10.93 12.51
2-Thienyl Phenyl
74
C
(512.60)
63.01 4.05 10.86 12.70
2-Thienyl 1-Naphthyl 78
C
31H22N4O3S2 66.17 3.94
9.96 11.40
(562.66)
65.98 4.07 10.07 11.32
2-Thienyl Phenyl
2-Thienyl Benzyl
2-Thienyl Benzoyl
70
68
66
72
74
76
80
C
27H22N4O2S3 61.11 4.18 10.56 18.13
(530.69)
61.05 4.22 10.43 18.02
C
28H24N4O2S3 61.74 4.44 10.29 17.66
(544.71)
61.80 4.36 10.37 17.46
C
28H22N4O3S3 60.19 3.97 10.03 17.22
(558.70)
59.98 4.07 10.12 17.15
3-Pyridyl
3-Pyridyl
3-Pyridyl
Phenyl
Benzyl
Benzoyl
182-4 C28H23N5O2S2 63.98 4.41 13.32 12.20
(525.65) 64.04 4.37 13.28 12.29
29H25N5O2S2 64.54 4.67 12.98 11.88
180-2
204-6
181-3
C
(539.67)
64.18 4.80 13.05 11.61
C
29H23N5O3S2 62.91 4.19 12.65 11.58
(553.66)
62.84 4.23 12.51 11.66
2-Thienyl Phenyl
C
29H22N4O3S3 61.03 3.89
9.82 16.86
9.91 16.78
(570.71)
60.95 3.99

Med Chem Res (2007) 16:300–318
313
Table 4 continued
Cpd,
R
R1 or R2
Y. (%) M.p.
(°C)
Mol. formula
(mol. weight)
Analysis
(calculated/found)
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
2-Thienyl Benzyl
2-Thienyl Benzoyl
76
72
76
75
78
79
78
74
75
78
76
79
76
72
77
200-2
160-2
C
30H24N4O3S3 61.62 4.14
9.58 16.45
9.66 16.38
9.36 16.70
9.50 16.65
(584.73)
61.55 4.23
C
30H22N4O4S3 60.18 3.70
(598.72)
60.25 3.58
3-Pyridyl
Phenyl
312-3 C30H23N5O3S2 63.70 4.10 12.38 11.34
(565.67) 63.48 4.18 12.30 11.71
2-Thienyl Phenyl
2-Thienyl Benzyl
2-Thienyl Benzoyl
157-9
198-9
212-4
[320
196-8
210-1
266-7
310-1
191-2
181-3
140-2
217-9
C
35H26N4O2S3 66.64 4.15
8.88 15.25
9.01 15.09
8.69 14.92
8.90 14.83
8.50 14.60
8.66 14.52
(630.80)
66.50 4.24
C
36H28N4O2S3 67.05 4.38
(644.83)
67.18 4.32
C
36H26N4O3S3 65.63 3.98
(658.81)
65.60 4.06
3-Pyridyl
Phenyl
C
36H27N5O2S2 69.10 4.35 11.19 10.25
(625.76) 69.21 4.29 10.99 10.33
2-Thienyl Phenyl
2-Thienyl Benzyl
2-Thienyl Benzoyl
C
30H24N4O3S3 61.62 4.14 9.58
16.45
9.70 16.38
9.36 16.07
9.32 16.19
9.14 15.70
9.22 15.46
(584.73)
61.55 4.20
C
31H26N4O3S3 62.18 4.38
(598.76)
62.05 4.41
C
31H24N4O4S3 60.76 3.95
(612.74)
60.51 4.06
3-Pyridyl
Phenyl
C
31H25N5O3S2 64.23 4.35 12.08 11.06
(579.69) 64.12 4.60 12.12 11.15
2-Thienyl Phenyl
2-Thienyl Benzyl
2-Thienyl Benzoyl
C
30H24N4O3S3 61.62 4.14
9.58 16.45
9.45 16.62
9.36 16.07
9.41 16.00
9.14 15.70
9.20 15.78
(584.73)
61.50 4.27
C
31H26N4O3S3 62.18 4.38
(598.76)
62.06 4.29
C
31H24N4O4S3 60.76 3.95
(612.74)
60.85 3.88
3-Pyridyl
Phenyl
C
31H25N5O3S2 64.23 4.35 12.08 11.06
(579.69) 64.33 4.28 11.95 11.30
H), 5.63 (m, 1H, C3-H), 7.12-8.01 (m, 11H, Ar-H). For 11: 2.20 (d, J = 9 Hz, 2H,
C4-2H), 2.73 (m, 1H, C3a-H), 5.42 (m, 1H, C3-H), 6.48–8.12 (m, 17H, Ar-H).
N1-Substituted N3-[4-(3-(2-thienyl)-3H-indeno[1,2-c]pyrazol-2-yl)-
benzenesulfonyl]-ureas (13–15).
To a stirred suspension of the appropriate sulfonylurea 7–9 (0.01 mol) in water (10
mL), bromine water (5%, 15 mL) was gradually added over a period of 30 min at

314
Med Chem Res (2007) 16:300–318
room temperature. After further stirring for 6h, the oxidation product thus formed,
was treated as described under 5 and 6. Physicochemical and analytical data are
recorded in Table 4. IR (cm–1): 1305–1320 and 1160–1170 (SO2N), 1650–1665
1
(C=O). H-NMR (d, ppm) for 14: 4.42 (s, 2H, C4-2H), 6.99-8.17 (m, 16H, Ar-H),
9.56 (s, 1H, NH), 10.31 (s, 1H, NH).
N1-Substituted N3-[4-(3-substituted-3a,4-dihydro-3H-indeno[1,2-c]pyrazol-2-
yl)-benzenesulf-onyl]-thioureas (16–21)
A solution of the appropriate isothiocyanate (0.011 mol) in dry acetone (5 mL) was
added to a stirred mixture of the appropriate sulfonamide 3 or 4 (0.01 mol) and
anhydrous potassium carbonate (2.8 g, 0.02 mol) in dry acetone (25 mL), and the
reaction mixture was heated under reflux with stirring for 10 h. The solvent was
removed under reduced pressure and the remaining solid residue was dissolved in
water (30 mL) and acidified with 2N hydrochloric acid. The precipitated crude
product was filtered, washed with water, dried, and recrystallized from ethanol.
Physicochemical and analytical data are recorded in Table 4. IR (cm–1): 1300–1335
and 1165–1200 (SO2N), 1155–1170 (C=S). For compounds 18 and 21 (R2
=
1
benzoyl): 1660–1675 (C=O). H-NMR (d, ppm) for 16: 2.22 (d, J = 9 Hz, 2H, C4-
2H), 2.61 (m, 1H, C3a-H), 5.49 (m, 1H, C3-H), 7.01–8.15 (m, 16H, Ar-H), 9.90 (s,
1H, NH), 10.12 (s, 1H, NH); for 17: 2.24 (d, J = 9 Hz, 2H, C4-2H), 2.58 (m, 1H,
C3a-H), 4.77 (s, 2H, benzyl-CH2), 5.50 (m, 1H, C3-H), 7.19–7.84 (m, 16H, Ar-H),
9.95 (s, 1H, NH), 10.30 (s, 1H, NH); for 19: 2.13 (d, J = 9 Hz, 2H, C4-2H), 2.65 (m,
1H, C3a-H), 5.42 (m, 1H, C3-H), 6.55–8.42 (m, 17H, Ar-H), 9.90 (s, 1H, NH), 10.04
(s, 1H, NH);
3-Substituted-2-[4-(3-substituted-3a,4-dihydro-3H-indeno[1,2-c]pyrazol-2-yl)-
benzenesulfo-nylimino]-thiazolidin-4-ones (22–25)
To a solution of the appropriate sulfonylthioureido derivative 16–19 (0.005 mol)
in absolute ethanol (20 mL) was added ethyl bromoacetate (1 g, 0.0055 mol) and
anhydrous sodium acetate (0.82 g, 0.01 mol) and the reaction mixture was heated
under reflux for 4 h. The mixture was left to attain room temperature then poured
into an ice-cold water (30 mL), and the resulting solid product was filtered,
washed with water, dried, and recrystallized from ethanol-benzene (1:1).
Physicochemical and analytical data are recorded in Table 4. IR (cm–1): 1720–
1
1735 (C=O). For compound 24 (R2 = benzoyl): 1690 (C=O). H-NMR (d, ppm)
for 22: 2.26 (d, J = 9 Hz, 2H, C4-2H), 2.66 (m, 1H, C3a-H), 4.18 (s, 2H, thiazol-
CH2), 5.56 (m, 1H, C3-H), 7.10–7.95 (m, 16H, Ar-H); for 25: 2.23 (d, J = 9 Hz,
2H, C4-2H), 2.57 (m, 1H, C3a-H), 3.98 (s, 2H, thiazol-CH2), 5.49 (m, 1H, C3-H),
6.88–8.54 (m, 17H, Ar-H).

Med Chem Res (2007) 16:300–318
315
3-Substituted-4-phenyl-2-[4-(3-substituted-3a,4-dihydro-3H-indeno[1,2-
c]pyrazol-2-yl)-benz-enesulfonylimino]-1,3-thiazolines (26–29)
A solution of the appropriate sulfonylthioureido derivative 16–19 (0.005 mol) in
absolute ethanol (20 mL) was refluxed with phenacyl bromide (1.1 g, 0.0055 mol)
and anhydrous sodium acetate (0.82 g, 0.01 mol) for 4 h during which the solid
product was partially crystallized out. The mixture was left to attain room
temperature then filtered, washed with cold ethanol, dried, and recrystallized from
ethanol. Physicochemical and analytical data are recorded in Table 4. IR (cm–1):
1
1600–1620 (C=N). For compound 28 (R2 = benzoyl): 1700 (C=O). H-NMR (d,
ppm) for 27: 2.16 (d, J = 9 Hz, 2H, C4-2H), 2.58 (m, 1H, C3a-H), 4.69 (s, 2H,
benzyl-CH2), 5.45 (m, 1H, C3-H), 6.07 (s, 1H, thiazol-H), 7.14–7.89 (m, 21H,
Ar-H); for 29: 2.20 (d, J = 9 Hz, 2H, C4-2H), 2.47 (m, 1H, C3a-H), 5.50 (m, 1H,
C3-H), 6.12 (s, 1H, Thiazol-H), 6.92–8.45 (m, 22H, Ar-H);
3-Substituted-2-[4-(3-substituted-3a,4-dihydro-3H-indeno[1,2-c]pyrazol-2-yl)-
benzenesulfon-ylimino]-[1,3]thiazinan-4-one (30–33)
To a solution of the appropriate thioureido derivative 16–19 (0.01 mol) in absolute
ethanol (20 mL) was added ethyl b-bromopropionate (0.011 mol) and sodium
acetate (0.2 mol) and the reaction mixture was heated under reflux for 6 h. Work up
of the reaction mixture was carried out as described for compounds 22–26.
Physicochemical and analytical data are recorded in Table 4. IR cm–1: 1720–1730
(C=O). For 28 (R2 = benzoyl): 1710 (C=O). 1H-NMR (d, ppm) for 30: 2.17 (d, J = 9
Hz, 2H, C4-2H), 2.58 (m, 1H, C3a-H), 3.30 (m, 4H, CH2-S + CH2-CO), 5.40 (m, 1H,
C3-H), 6.89–8.10 (m, 21H, Ar-H); for 33: 2.16 (d, J = 9 Hz, 2H, C4-2H), 2.36
(m, 1H, C3a-H), 3.40 (m, 4H, CH2-S + CH2-CO), 5.55 (m, 1H, C3-H), 6.78–8.36
(m, 22H, Ar-H).
3-Substituted-2-[4-(3-substituted-3a,4-dihydro-3H-indeno[1,2-c]pyrazol-2-yl)-
benzenesulfon-ylimino]-[1,3]thiazinan-5-one (34–37)
To a solution of the appropriate thioureido derivative 16–19 (0.01 mol) in absolute
ethanol (20 mL) was added 1,3-dichloroacetone (0.011 mol) and sodium acetate (25
mmol) and the reaction mixture was heated under reflux for 8 h. Work up of the
reaction mixture was carried out as described for compound 7. Physicochemical and
analytical data are recorded in Table 4. IR cm–1: 1720-1730 (C=O). For compound
36 (R2 = benzoyl): 1695 (C=O). 1H-NMR (d, ppm) for 34: 2.19 (d, J = 9 Hz, 2H, C4-
2H), 2.60 (m, 1H, C3a-H), 4.55 (s, 2H, CH2), 5.29 (s, 2H, CH2), 5.38 (m, 1H, C3-H),
6.82–8.22 (m, 21H, Ar-H); for 37: 2.16 (d, J = 9 Hz, 2H, C4-2H), 2.36 (m, 1H, C3a-
H), 4.62 (s, 2H, CH2), 5.18 (s, 2H, CH2), 5.47 (m, 1H, C3-H), 6.69–8.43 (m, 22H,
Ar-H).

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Med Chem Res (2007) 16:300–318
In Vitro Antitumor Screening
Compounds 3, 5, 11, 13, 14, 16, 18, 24, 26, 28, 30, 32 and 34 were evaluated for
their antitumor activity according to the protocol of the NCI in vitro disease-
oriented human cells screening panel assay (Grever et al. 1992; Boyd and Paull
1995; Monks et al. 1991).
Acknowledgement The authors are deeply thankful to the authorities of the Institute of Research and
Consultation, King Abdulaziz University, Jeddah, Kingdom Saudi Arabia, for financial support of this
research project (425/006). Thanks are extended to the staff members of the Department of Health and
Human Services, National Cancer Institute (NCI), Bethesda, Maryland, USA for carrying out the
anticancer screening of the newly synthesized compounds.
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