Synthesis and antitumor evaluation of novel sulfonylcycloureas derived from nitrogen mustard

Article abstract of DOI:10.1007/s11030-015-9647-6

A new series of sulfonylcycloureas derivatives have been synthesized and evaluated in vitro for their antitumor activity against four cancer cell lines (A431, Jurkat, U266, and K562). These compounds were prepared by the condensation of several sulfonamides (2am) with ethyl bis(2-chloroethyl)carbamate (1a). The relative cytotoxicity of these new derivatives in comparison to chlorambucil is reported.

Full text of DOI:10.1007/s11030-015-9647-6

Mol Divers
DOI 10.1007/s11030-015-9647-6
FULL-LENGTH PAPER
Synthesis and antitumor evaluation of novel sulfonylcycloureas
derived from nitrogen mustard
H. Cheloufi1 · B. Belhani1 · T. S. Ouk2 · R. Zerrouki2 · N.-E. Aouf1 ·
M. Berredjem1
Received: 28 May 2015 / Accepted: 2 November 2015
© Springer International Publishing Switzerland 2015
Abstract A new series of sulfonylcycloureas derivatives
have been synthesized and evaluated in vitro for their anti-
tumor activity against four cancer cell lines (A431, Jurkat,
U266, and K562). These compounds were prepared by the
condensation of several sulfonamides (2a–m) with ethyl
bis(2-chloroethyl)carbamate (1a). The relative cytotoxicity
of these new derivatives in comparison to chlorambucil is
reported.
countries [2]. Potent antitumor drugs, such as nitrogen mus-
tards (e.g., chlorambucil, melphalan, and mechlorethamine)
[3], have been widely used in cancer chemotherapy for many
years. These compounds were identified as DNA-targeted
alkylating agents due to formation of cross links within
the DNA chain of tumor cells. The development of these
compounds bears several challenges and has received much
attention in order to overcome drawbacks including high
chemical reactivity inducing bone marrow toxicity or reduc-
tion of cytotoxicity via DNA repair mechanism. In the last
decade, many structural modifications have been envisioned
to increase cytotoxicity, stability, and selectivity for targeting
the DNA of tumor cells [4–10].
Keywords Cancer · Sulfonamides · Nitrogen mustard ·
Sulfonylcycloureas · Antitumor
Introduction
After the discovery of Sulofenur (LY186641(A), Fig. 1)
an antineoplastic sulfonylurea [11], an extensive range of
novel compounds containing sulfonamides [12–19] and
diaryl sulfonylurea [20–25] moieties have attracted consid-
erable attention and been evaluated against various types
of cancer. Later, El-Deeb et al. synthesized several novel
cyclic arylsulfonylureas ((B), Fig. 1) and evaluated their
antitumor activity against different human solid tumors
[26]. In addition, other sulfonylurea derivatives with impor-
tant pharmaceutical activities, such as hypoglycemic ((C),
Fig. 1) [27], H3 receptor antagonists [28] ((D), Fig. 1),
and antimalarial [29] ((E), Fig. 1) activity, were recently
described.
Cancer is a major public health problem and a leading cause
of death worldwide [1]. More than half of all cancer cases
and 60 % of deaths from cancer appear in less developed
Electronic supplementary material The online version of this
article (doi:10.1007/s11030-015-9647-6) contains supplementary
material, which is available to authorized users.
In the current study, our aim is to explore new selective
DNA-directed alkylating agents, less toxic and more sta-
ble. We have synthesized a novel series of sulfonylcyclourea
derivatives (4a–m) containing a nitrogen mustard moiety
using commercially available chlorosulfonyl isocyanate and
N,N-bis(2-chloroethyl)amine as the starting material. These
new derivatives were evaluated for their in vitro antitumor
activity against four cancer cell lines (A431, Jurkat, U266,
and K562).
M. Berredjem
malika.berredjem@univ-annaba.org
B
1
Laboratory of Applied Organic Chemistry, Synthesis of
Biomolecules and Molecular Modelling Group,
Badji-Mokhtar, Annaba University, BOX 12, Annaba, Algeria
2
Laboratoire de Chimie des Substances Naturelles, Université
de Limoges, 123 Av. Albert Thomas, 87060 Limoges Cedex,
France
123

Mol Divers
bis(2-chloroethyl)carbamate (1a) sulfonamides (2a–m) in
acetonitrile using K2CO3 as a base under refluxing condi-
tions for 3–5 h generated intermediates (3a–m). The strong
acidity of the NH proton between SO2 and C=O facilitates
the intermolecular cyclization leading to the desired products
(4a–m) in good yields. The physical data and yields of the
final products are listed in Table 1.
Antitumor activity
The cytotoxic effects of the new products were determined in
T-lymphoma cells (Jurkat), Chronic Myelogenous Leukemia
cells (K562), Myeloma cells (U266), and Epidermoid carci-
noma cells (A431) by MTT assay. Chlorambucil was used as
reference. IC50 values were calculated by logistic regression
analysis of dose–response curves plotting between percent-
age of viability and the concentration of the test compounds.
The results are summarized in Table 2. Compounds 4c and
4d showed some cytotoxicity. For compound 4c, the IC50
value is 422 μM for Jurkat, 482 μM for K562, 908 μM for
U266, and 638 μM for A431. Interestingly, compound 4d
displayed higher cytotoxicity than 4c. For this compound,
the IC50 value is 390 μM, 464 μM, 473 μM, and 315 μM,
respectively, for Jurkat, K562, U266, and A431. Further
investigations are currently in progress to evaluate the impact
on cancer cell cycle by these structures and the cell death
pathway involved.
Fig. 1 Examples of bioactive sulfonylurea derivatives
Results and discussion
Chemistry
Our synthetic route for the preparation of sulfonylcyclourea
derivatives (4a–m) is outlined in Scheme 1. The synthe-
sis of sulfonamide derivatives (2a–m) [30–35] was carried
out in three steps, by carbamoylation of chlorosulfonyl iso-
cyanate with tert-butanol in anhydrous conditions to form
the N-chlorosulfonyl carbamate, which was then reacted
with various amines (sulfamoylation). Removal of the tert-
butyloxycarbonyl(BOC)protectinggroupwasaccomplished
under mild conditions in water according to a previously
described procedure [36]. Ethyl bis(2-chloroethyl)carbamate
(1a) was prepared in excellent yield by direct acylation of
commercially available N,N-bis(2-chloroethyl)amine with
ethyl chloroformate in the presence of triethylamine in anhy-
drous dichloromethane at 0 ◦C. The condensation of ethyl
Conclusion
In the present work, we have developed the synthesis of
a new series of sulfonylcyclourea derivatives containing a
nitrogen mustard moiety, starting from commercially avail-
able compounds. The antitumor activity of these molecules
was evaluated against 4 different cancer cell lines (Jurkat,
K562, U266, and A431) using Chlorambucil as reference.
Compound 4d showed higher cytotoxicity than compound
4c; however, when compared to chlorambucil, its biological
activity remained modest.
Experimental procedure
Chemistry
Chemical materials, solvents, and reagents were reagent
grade and were used without purification. Melting points
were recorded using a Büchi® melting point apparatus Model
B-540 and are uncorrected. Reaction progress was mon-
itored using analytical thin-layer chromatography (TLC)
on 0.25-mm Merck F-254 silica gel glass plates. Column
Scheme 1 Synthesis of novel sulfonylcyclourea derivatives (4a–m)
123

Mol Divers
Table 1 Physical properties, yields, and molecular structures of products (4a–m)
Entry
R
Compound
Time(hours) Yield (%)
m.p. (°C)
H
H
N
N
N
N
l
C
S
3
86
83
90–92
96–98
4a
4b
O
O
O
H
N
H
3.5
N
N
N
l
C
S
O
O
O
H
N
H
N
N
N
5
3
90
88
83–84
4c
l
C
S
O
O
O
O
H
N
H
N
F
N
F
N
l
C
S
101–103
4d
O
O
H
N
H
N
N
N
l
C
3.5
5
81
77
99–101
91–92
4e
4f
S
O
O
O
l
C
l
C
H
N
H
N
N
N
l
C
S
O
O
e
M
O
O
N
e
M
O
4.5
3.5
73
82
100–102
111–113
4g
4h
e
M
e
M
O
O
H
H
N
N
N
l
C
S
O
O
O
N
N
N
N
l
C
S
O
O
O
O
O
N
N
N
5
4
89
92
104–106
110–111
4i
4j
l
C
S
N
O
O
O
N
N
N
N
N
N
l
C
S
O
O
O
4
90
101–102
4k
N
N
N
N
l
C
S
N
O
O
O
H
N
H
N
N
l
C
S
5
5
87
88
97–99
4l
O
O
O
H
H
N
N
N
N
l
C
S
98–100
4m
O
O
O
123

Mol Divers
Table 2 Antitumor profile of compounds (4c, 4d)
IC50 (μM)
General procedure for the synthesis of sulfonylcyclourea
derivatives (4a–m)
A431
Jurkat
U266
K562
A mixture of ethyl bis(2-chloroethyl)carbamate (1a) and 1
equiv of sulfonamides derivatives (2a–m) in the presence of
K2CO3 (1.5 equiv) was heated to reflux in acetonitrile under
stirring for 3–5 h. The progress of the reaction was monitored
by TLC using dichloromethane and methanol (9.9:0.1). The
reaction mixture was evaporated and diluted with methylene
chloride (30 mL) and then washed with water. The organic
layer was dried over Na2SO4 and concentrated in vacuo.
The residue was purified by silica gel chromatography using
CH2Cl2 as eluent.
Chlorambucil
68
638
315
113
422
390
143
908
473
159
482
464
4c
4d
chromatography was performed using silica gel 60 Å (63–
200 μm). 1H NMR spectra were recorded on a Bruker AM
300 spectrometer and chemical shifts were reported in parts
per million (ppm) using tetramethylsilane as internal stan-
dard with coupling constants (J) reported in Hertz (Hz).
NMR signals are denoted as follows: s, singlet; d, doublet;
t, triplet; q, quartet; m, multiplet. Elemental analyses were
performed using a Perkin–Elmer 2400 C, H, N analyzer, and
the determined values were within the acceptable limits of
the calculated values. IR spectra were recorded on a Perkin
Elmer Spectrum Version10.4.1 RX-I spectrometer using KBr
disks.
Bis(2-chloroethyl)carbamate (1a) Yellow oil; 99 % Yield;
Rf = 0.6 (CH2Cl2/MeOH, 9:1); IR (KBr, cm−1) : 1633
ν
1
(C=O); H NMR (400 MHz, CDCl3) δ: 4.15 (q, 2H, J =
6.03, CH2–O), 3.69–3.50 (m, 8H, 2(2CH2–Cl)), 1.22 (t, 3H,
J = 5.95, CH3–CH2); 13C NMR (100 MHz, CDCl3) δ:
155.8, 61.8, 51.0, 41.9, 14.5; ESI-MS m/z: 214 [M+1]+;
Anal Calcd for C13H18ClN3O3S: C 39.27, H 6.12, N 6.54;
found: C 39.15, H 6.09, N 6.58.
N-chloroethyl,Nꢀ-(1-phenyl)sulfamoyl,imidazolidine-2-one
(4a)Whitepowder; 86%Yield; Rf = 0.63(CH2Cl2/MeOH,
General procedure for the synthesis of ethyl
bis(2-chloroethyl)carbamate (1a)
9:1); m.p. 90–92 ◦C, IR (KBr, cm−1) : 3228 (NH), 1630
ν
(C=O), 1327, 1160 (O=S=O); 1H NMR (400 MHz, CDCl3)
δ: 7.05–7.37 (m, 5H, Ar–H), 6.70 (S, 1H, NH), 4.60 (t,
J = 8.24 Hz, 2H, Cl–CH2), 3.90 (t, J = 7.20 Hz, 2H,
NH–CH2), 3.57–3.61 (m, 4H, (CH2)cyc); 13C NMR (100
MHz, CDCl3) δ: 161.5, 140.3, 129.3, 123.7, 121.8, 68.9,
49.0, 47.0; ESI-MS m/z: 304 [M+1]+; Anal Calcd for
C11H14ClN3O3S: C 43.49, H 4.65, N 13.83, S 10.56; found:
C 43.41, H 4.59, N 13.87, S 10.44.
The N-acylation reaction of bis(2-chloroethyl)amine (0.5 g,
2.8 mmol) with ethyl chloroformate (0.45 g, 0.39 mL, 1.5
equiv) was carried out in anhydrous dichloromethane at 0 ◦C
for 1.5 h in the presence of triethylamine (1.5 equiv). The
organic phase was washed with HCl (0.1 N), then with water,
and then dried over Na2SO4. After evaporation of the solvent
under reduced pressure, the product was obtained as an oil
in excellent yield.
N-chloroethyl,Nꢀ-(1-benzyl)sulfamoyl,imidazolidine-2-one
(4b)Whitepowder;83%Yield; Rf = 0.65(CH2Cl2/MeOH,
9:1); m.p. 96–98 ◦C; IR (KBr, cm−1) : 3237 (NH), 1641
ν
(C=O), 1335, 1171 (O=S=O); 1H NMR (400 MHz, CDCl3)
δ: 7.11–7.43 (m, 5H, Ar–H), 6.77 (S, 1H, NH), 4.61 (t,
J = 8.24 Hz, 2H, Cl–CH2), 3.92 (t, J = 7.20 Hz, 2H,
NH–CH2), 3.62–3.59 (m, 4H, (CH2)cyc), 4.04 (d, J = 6.1,
CH2–Ph); 13C NMR (100 MHz, CDCl3) δ: 159.4, 143.3,
128.3, 121.7, 119.8, 68.9, 49.3, 46.9, 43.5; ESI-MS m/z:
318 [M+1]+; Anal Calcd for C12H16ClN3O3S: C 45.35, H
5.07, N 13.22, S 10.09; found: C 45.29, H 5.03, N 13.16, S
10.11.
General procedure for the synthesis of sulfonamide
derivatives (2a–m)
Sulfonamidederivativeswereobtainedbyslowlyaddingtert-
butanol (0.85 g, 1.09 mL, 11.48 mmol) to a stirred solution of
chlorosulfonyl isocyanate (CSI) (1.62 g, 1 mL, 11.48 mmol)
in anhydrous methylene chloride (10 mL) at 0 ◦C. After 30
min, the resulting solution of N-chlorosulfonyl carbamate
(1.75 mL, 1.1 equiv.) was slowly added to a solution of pri-
mary amine (1 equiv) in anhydrous methylene chloride (10
mL) at 0 ◦C in the presence of triethylamine. Stirring was
continued for 2 h. The reaction mixture was diluted with
methylene chloride (30 mL) and subsequently washed with
0.1 N HCl (20 mL) and then with water (25 mL). The organic
layer was dried over anhydrous sodium sulfate and concen-
trated in vacuo to give the desired sulfonamide derivatives
(2a–m) in excellent yields.
N-chloroethyl,Nꢀ-(1-methylbenzyl)sulfamoyl,imidazolidine-
2-one (4c) White powder; 90 % Yield; Rf = 0.61 (CH2Cl2/
MeOH, 9:1); m.p. 83–84 ◦C; IR (KBr, cm−1) : 3230 (NH),
ν
1
1639 (C=O), 1311, 1142 (O=S=O); H NMR (250 MHz,
CDCl3) δ: 7.48–7.22 (m, 5H, Ar–H), 4.70 (t, J = 7.39,
1H, CH2),4.50 (t, J = 7.46, 1H, CH2), 4.40 (t, J =
7.71, 1H, CH2), 4.20 (t, J = 8.12, 1H, CH2), 3.70 (q,
123

Mol Divers
J = 7.38 Hz, 1H, *CH–Ph), 3.60–3.20 (m, 4H, (CH2)cyc),
1.4 (d, J = 6.9 Hz, 3H, CH3–*CH); 13C NMR (60 MHz,
CDCl3) δ: 164.1, 141.9, 128.2, 127.1, 125.6, 66.1, 53.4,
47.1, 46.9, 40.3; ESI-MS m/z: 332 [M+1]+; Anal Calcd for
C13H18ClN3O3S: C 47.06, H 5.47, N 12.66, S 9.66; found:
C 47.02, H 5.44, N 12.61, S 9.60.
for C12H16ClN3O4S: C 43.18, H 4.83, N 12.59, O 19.17, S
9.61; found: C 43.23, H 4.80, N 12.53, S 9.58.
N-chloroethyl,Nꢀ-(1-diphenyl)sulfamoyl,imidazolidine-2-
one (4h) White powder; 82 % Yield; Rf = 0.71 (CH2Cl2/
MeOH, 9:1); m.p. 111–113 ◦C; IR (KBr, cm−1) : 3235
ν
1
(NH), 1635 (C=O), 1340, 1180 (O=S=O); H NMR (400
MHz, CDCl3) δ: 7.17–6.65 (m, 10H, Ar–H), 4.57 (t,
J = 8.01 Hz, 2H, Cl–CH2), 3.88 (t, J = 6.66 Hz, 2H,
NH–CH2), 3.49–3.57 (m, 4H, (CH2)cyc); 13C NMR (100
MHz, CDCl3) δ: 165.2, 139.5, 128.4, 124.1, 122.3, 69.1,
49.5, 47.1; ESI-MS m/z: 380 [M+1]+; Anal Calcd for
C17H18ClN3O3S: C 53.75, H 4.78, N 11.06, S 8.44; found:
C 53.70, H 4.81, N 11.00, S 8.49.
N-chloroethyl,Nꢀ-(3-fluorophenyl)sulfamoyl,imidazolidine-
2-one (4d) White powder; 88 % Yield; Rf = 0.66 (CH2Cl2/
MeOH, 9:1); m.p. 101–103 ◦C; IR (KBr, cm−1) : 3219
ν
1
(NH), 1620 (C=O), 1322, 1159 (O=S=O); H NMR (400
MHz, CDCl3) δ: 7.20–7.03 (m, 4H, Ar–H), 4.55 (t, J =
6.23 Hz, 2H, Cl–CH2), 3.85 (t, J = 6.50 Hz, 2H, N-
CH2), 3.60–3.51 (m, 4H, (CH2)cyc); 13C NMR (100 MHz,
CDCl3) δ: 160.5, 140.1, 136.2, 123.7, 114.5, 66.1, 47.0, 46.9;
N-chloroethyl,Nꢀ-(morpholin-4-yl)sulfamoyl,imidazolidine
ESI-MSm/z:322[M+1]+;AnalCalcdforC11H13ClFN3O3S: -2-one (4i) White powder; 89 % Yield; Rf
=
0.71
ν
C 41.06, H 4.07, N 13.06, S 9.97; found: C 41.00, H 4.12, N
13.03, S 9.92.
(CH2Cl2/MeOH, 9:1); m.p. 104–106 ◦C; IR (KBr, cm−1) :
1
1611 (C=O), 1338, 1178 (O=S=O); H NMR (400 MHz,
CDCl3) δ: 4.64 (t, J = 8.46 Hz, 2H, Cl–CH2), 3.90 (t,
J = 7.17 Hz, 2H, N–CH2), 3.77 (t, J = 4.50Hz, 4H, 2CH2–
O), 3.71–3.67 (m, 4H, (CH2)cyc), 3.16 (t, J = 4.71 Hz, 4H,
2CH2–N); 13C NMR (100 MHz, CDCl3) δ: 163.4, 66.9, 65.0,
49.0, 47.0, 44.2; ESI-MS m/z: 298 [M+1]+; Anal Calcd for
C9H16ClN3O4S: C 36.30, H 5.42, N 14.11, S 10.77; found:
C 36.26, H 5.48, N 14.09, S 10.63.
N-chloroethyl,Nꢀ-(4-chlorophenyl)sulfamoyl,imidazolidine-
2-one (4e) White powder; 81 % Yield; Rf = 0.63 (CH2Cl2/
MeOH, 9:1); m.p. 99–101 ◦C; IR (KBr, cm−1) : 3226 (NH),
ν
1
1619 (C=O), 1315, 1149 (O=S=O); H NMR (400 MHz,
CDCl3) δ: 7.22–7.12 (m, 4H, Ar–H), 4.62 (t, J = 6.23 Hz,
2H, Cl–CH2), 3.81 (t, J = 6.50 Hz, 2H, N–CH2), 3.66–3.58
(m, 4H, (CH2)cyc); 13C NMR (100 MHz, CDCl3) δ: 162.5,
139.1, 131.2, 124.7, 113.5, 68.1, 49.0, 45.9; ESI-MS m/z:
338 [M+1]+; Anal Calcd for C11H13Cl2N3O3S: C 39.06, H
3.87, N 12.42, S 9.48; found: C 39.10, H 3.82, N 12.39, S
9.52.
N-chloroethyl,Nꢀ-(4-phenylpiperazin-1-yl)sulfamoyl,imida
zolidine-2-one (4j) Yellow powder; 92 % Yield; Rf = 0.71
(CH2Cl2/MeOH, 9:1); m.p. 110–111 ◦C; IR (KBr, cm−1) :
ν
1
1629 (C=O), 1320, 1166 (O=S=O); H NMR (400 MHz,
CDCl3) δ: 6.85–7.34 (m, 5H, Ar–H), 4.68 (t, J = 8.2 Hz,
2H, Cl–CH2), 3.87 (t, J = 8.16Hz, 2H, N–CH2), 3.70–3.65
(m, 4H, (CH2)cyc), 3.35–3.25 (2m, 8H, 2(N–CH2– CH2–
N)); 13C NMR (100 MHz, CDCl3) δ: 160.3, 137.2, 129.3,
117.2, 116.7, 66.9, 53.1, 49.0, 47.0, 44.8; ESI-MS m/z: 373
[M+1]+; Anal Calcd for C15H21ClN4O3S: C 48.32, H 5.68,
N 15.03, S 8.60; found: C 48.27, H 5.71, N 15.09, S 8.54.
N-chloroethyl,Nꢀ-(4-methoxyphenyl)sulfamoyl,imidazolid
ine-2-one (4f) White powder; 77 % Yield; Rf = 0.66
(CH2Cl2/MeOH, 9:1); m.p. 91–92 ◦C; IR (KBr, cm−1) :
ν
1
3239 (NH), 1627 (C=O), 1330, 1169 (O=S=O); H NMR
(400 MHz, CDCl3) δ: 7.20 (d, J = 8.8 Hz, 2H, Ar–H),
6.85 (d, J = 8.8 Hz, 2H, Ar–H), 4.60 (t, J = 6.23 Hz,
2H, Cl–CH2), 3.85 (t, J = 6.50 Hz, 2H, N–CH2), 3.75(s,
1H, CH3–O), 3.60–3.55 (m, 4H, (CH2)cyc); 13C NMR (100
MHz, CDCl3) δ: 161.5, 140.2, 135.1, 123.7, 114.5, 67.1,
47.2, 47.05, 41.5;ESI-MS m/z:334[M+1]+;AnalCalcdfor
C12H16ClN3O4S: C 43.18, H 4.83, N 12.59, S 9.61; found:
C 43.22, H 4.79, N 12.52, S 9.55.
N-chloroethyl,Nꢀ-(1,2,3,4-tetrahydroisoquinolin2-yl)sulf
amoyl,imidazolidine-2-one (4k) White powder;90 % Yield;
Rf = 0.71 (CH2Cl2/MeOH, 9:1); m.p. 101–102 ◦C; IR
1
(KBr, cm−1) : 1632 (C=O), 1302, 1150 (O=S=O); H NMR
ν
(400 MHz, CDCl3) δ: 7.18–7.05 (m, 4H, Ar–H), 4.60 (t,
J = 8.28 Hz, 2H, Cl–CH2), 4.40 (s, 1H, Ph–CH2–N),
3.80 (t, J = 7.14 Hz, 2H, N–CH2), 3.60–3.45 (m, 6H,
(CH2)cyc + CH2–N), 2.97 (t, J = 6.03 Hz, 2H, Ph–CH2);
13C NMR (100 MHz, CDCl3) δ: 162.3, 134.2, 127.5, 126.5,
67.9, 49.6, 48.0, 46.5, 41.7, 25.3;ESI-MS m/z:344 [M+1]+;
Anal Calcd for C14H18ClN3O3S: C 48.91, H 5.28, N 12.22,
S 9.33; found: C 49.00, H 5.32, N 12.16, S 9.25.
N-chloroethyl,Nꢀ-(2-methoxyphenyl)sulfamoyl,imidazolid
ine-2-one (4g) White powder; 73 % Yield; Rf = 0.66
◦
(CH2Cl2/MeOH, 9:1); m.p. 100–102 C; IR (KBr, cm−1) :
ν
1
3230 (NH), 1625 (C=O), 1325, 1163 (O=S=O); H NMR
(400 MHz, CDCl3) δ: 7.25 (d, J = 8.1 Hz, 2H, Ar–H),
6.99 (d, J = 8.1 Hz, 2H, Ar–H), 4.53 (t, J = 7.10 Hz,
2H, Cl–CH2), 3.99 (t, J = 6.20 Hz, 2H, N–CH2), 3.81(s,
1H, CH3–O), 3.65–3.59 (m, 4H, (CH2)cyc); 13C NMR (100
MHz, CDCl3)δ: 165.0, 143.4, 133.5, 121.8, 112.3, 65.7,
49.3, 46.9, 39.9; ESI-MS m/z: 334 [M+1]+; Anal Calcd
N-chloroethyl,Nꢀ-(1-propyl)sulfamoyl,imidazolidine-2-one
(4l)Yellowpowder;87%Yield; Rf = 0.71(CH2Cl2/MeOH,
9:1); m.p. 97–99 ◦C; IR (KBr, cm−1) : 3249 (NH), 1615
ν
123

Mol Divers
(C=O), 1307, 1140 (O=S=O); 1H NMR (400 MHz, CDCl3)
δ: 4.65 (t, J = 7.35 Hz, 2H, Cl–CH2), 4.22 (t, J =
6.03 Hz, 1H, NH), 3.71–3.60 (m, 4H, (CH2)cyc), 3.08 (q,
J = 6.96 Hz, 2H, CH2–NH), 1.57 (m, 2H, CH2), 0.94 (t,
J = 7.35 Hz, 3H, CH3); 13C NMR (100 MHz, CDCl3) δ:
160.9, 67.1, 47.0, 46.8, 45.0, 41.3, 22.0, 10.0; ESI-MS m/z:
270 [M+1]+; Anal Calcd for C8H16ClN3O3S: C 35.62, H
5.98, N 15.58, S 11.89; found: C 35.57, H 6.01, N 15.50, S
11.93.
incubated for 3 h at 37 ◦C. The purple formazan crystals were
dissolved by adding 200 μL DMSO. The absorbance was
read using a microplate spectrophotometer (Triad, Dynex
Technologies) at 595 nm. All measurements were carried
in triplicates. The results were compared with those of a
control reference plate fixed on the treatment day, and the
growth inhibition percentage was calculated for each drug
contact period. The concentration required for 50 % inhibi-
tion of cell viability (IC50) was calculated using the software
Origin Pro (Origin Lab, Northampton, USA). Assays were
performed in hexaplicate on three independent experiments.
N-chloroethyl,Nꢀ-(1-ethyl)sulfamoyl,imidazolidine-2-one
(4m) Yellow powder; 88 % Yield; Rf = 0.66 (CH2Cl2/
◦
MeOH, 9:1);m.p. 98–100 C;IR(KBr, cm−1) :3240(NH),
ν
Acknowledgments This work was supported financially by The
General Directorate for Scientific Research and Technological Devel-
opment (DG-RSDT), Algerian Ministry of Scientific Research, Applied
Organic Laboratory (FNR 2000). We also thank Pr. Jacques Lebreton
from the University of Nantes, Dr. Zouhair Bouaziz and Pr. Marc Le
Borgne from the university Claude Bernard Lyon for their help in the
identification for all products in NMR and MS.
1
1622 (C=O), 1310, 1146 (O=S=O); H NMR (400 MHz,
CDCl3) δ: 4.70 (t, J = 7.20 Hz, 2H, Cl–CH2), 4.15 (t,
J = 6.22 Hz, 1H, NH), 3.85–3.71 (m, 4H, (CH2)cyc), 2.88
(m, J = 7.36 Hz, 2H, NH–CH2), 1.01 (t, J = 7.35 Hz, 3H,
CH3); 13C NMR (100 MHz, CDCl3) δ: 162.1, 68.3, 47.5,
46.4, 44.9, 40.8, 12.5; ESI-MS m/z: 256 [M+1]+; Anal
CalcdforC7H14ClN3O3S:C32.88, H5.52, N16.43, S12.54;
found: C 32.93, H 5.46, N 16.38, S 12.49.
References
1. Eckhardt S (2002) Recent progress in the development of anti-
cancer agents. Curr Med Chem-Anti-Cancer Agents 2:419–439.
doi:10.2174/1568011024606389
Antitumor activity
2. Boyle P, Levin B (2008) World cancer report, the World Health
Organization’s International Agency for Research on Cancer
(IARC) Lyon, pp. 16025, 40, 42
Cell lines and culture conditions
3. Francisco AP, Perry MJ, Moreira R, Mendes E (2008) Chapter 9.
Alkylating agents. In: Misssailidis S (ed) Anticancer therapeutics.
Wiley, New York, pp 133–158
Jurkat (acute T cell Leukemia, ATCC TIB-152), K562
(chronic myelogenous leukemia, ATCC CLL-243) and U266
(multiple myeloma, ATCC TIB-196) were routinely cul-
tured in RPMI-1640 supplemented with 10 % Fetal Bovine
Serum (FBS, IDBIO, Limoges, France), 100 U/mL peni-
cillin, 100 μg/mL streptomycin, 2 mM l-glutamine, 1 mM
sodium pyruvate, 1 % vitamins, and 1 % non-essentials
amino-acids. A431 (vulvar epidermoid carcinoma, ATCC
CRL-1555) was cultured in DMEM supplemented with 10 %
FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, 2 mM
l-glutamine, and 1 mM sodium pyruvate. The cells were cul-
tured at 37 ◦C in a fully humidified 5 % CO2 incubator. All
reagents for cell culture were purchased from Gibco-BRL
Life Technologies, Cergy-Pontoise, France.
4. Kapuriya N, Kakadiya R, Dong H, Kumar A, Lee PC, Zhang X,
Chou TC, Lee TC, Chen CH, Lam K, Marvania B, Shah A, Su
TL (2011) Design, synthesis, and biological evaluation of novel
water-soluble N-mustards as potential anticancer agents. Bioorg
Med Chem 19:471–485. doi:10.1016/j.bmc.2010.11.005
5. Cimino P, Bifulco G, RiccioR Gomez-Palomab L, Barone V (2006)
On the role of stereo-electronic effects in tuning the selectivity
and rate of DNA alkylation by duocarmycins. Org Biomol Chem
4:1242–1251. doi:10.1039/B514890A
6. Leiris S, Lucas M, Dupuyd’Angeac A, Morere A (2010) Synthesis
and biological evaluation of cyclic nitrogen mustards based on car-
nitine framework. Eur J Med Chem 45:4140–4148. doi:10.1016/j.
ejmech.2010.06.003
7. Seifert J, Pezeshki S, Kamal A, Weisz K (2012) Inter- and
intrastrand DNA crosslinks by 2-fluoro-substituted pyrrolobenzo-
diazepine dimers: stability, stereochemistry and drug orientation.
Org Biomol Chem 10:6850–6860. doi:10.1039/C2OB25654A
8. Li S, Wang X, He Y, Zhao M, Chen Y, Xu J, Feng M, Chang J, Ning
H, Qi C (2013) Designing and synthesis of novel quinazoline nitro-
gen mustard derivatives as potential therapeutic agents for cancer.
Eur J Med Chem 67:293–301. doi:10.1016/j.ejmech.2013.06.055
9. Mourelatos C, Kareli D, Dafa E, Argyraki M, Koutsourea A,
Papakonstantinou I, Fousteris M, Pairas G, Nikolaropoulos S,
Lialiaris TS (2012) Cytogenetic and antineoplastic effects by newly
synthesised steroidal alkylators in lymphocytic leukaemia P388
cells in vivo. Mutat Res 746:1–6. doi:10.1016/j.mrgentox.2011.
12.032
Cell viability assay
Cytotoxicity activity was evaluated using the MTT assay
(3-(4,5-dimethylthiazol-2-yl)-phenyl-tetrazolium bromide).
Briefly, tumor cell lines were added into 96-well tissue cul-
ture plates in culture medium. Compounds were prepared at
a concentration of 4000 μM in 10 % DMSO in complete cul-
ture medium (v/v). The solutions were used at concentration
range from 2000 μM to 3.9 μM. Cells were incubated with
or without drugs for 72 h. Then, the MTT solution was added
at a final concentration of 0.5 mg/mL per well and cells were
10. Scutaru AM, Wenzel M, Gust R (2011) Bivalent bendamustine
and melphalan derivatives as anticancer agents. Eur J Med Chem
46:1604–1615. doi:10.1016/j.ejmech.2011.02.008
123

Mol Divers
11. Howbert JJ, Grossman CS, Crowell TA, Rieder BJ, Harper RW,
Grindey GB (1990) Novel agents effective against solid tumors: the
diaryl sulfonylureas. Synthesis, activities, and analysis of quanti-
tative structure-activity relationships. J Med Chem 33:2393–2407.
doi:10.1021/jm00171a013
12. Crespo R, De Bravo MG, Colinas PA, Bravo RD (2010) In vitro
antitumor activity of N-glycosyl sulfonamides. Bioorg Med Chem
Lett 20:6469–6471. doi:10.1016/j.bmcl.2010.09.052
13. Mojzycha M, Bielawska A, Bielawski K, Ceruso M, Supuran
CT (2014) Pyrazolo[4,3-e][1,2,4]triazine sulfonamides as carbonic
anhydrase inhibitors with antitumor activity. Bioorg Med Chem
22:2643–2647. doi:10.1016/j.bmc.2014.03.029
14. El-Sayed NS, El-Bendary ER, El-Ashry SM, El-Kerdawy MM
(2011) Synthesis and antitumor activity of new sulfonamide
derivatives of thiadiazolo[3,2-a] pyrimidines. Eur J Med Chem
46:3714–3720. doi:10.1016/j.ejmech.2011.05.037
15. Zhu M, Zhang C, Nwachukwu JC, Srinivasan S, Cavett V, Zheng
Y, Carlson KE, Dong C, Katzenellenbogen JA, Nettleand KW,
Zhou HB (2012) Bicyclic core estrogens as full antagonists: syn-
thesis, biological evaluation and structure-activity relationships
of estrogen receptor ligands based on bridged oxabicyclic core
arylsulfonamides. Org Biomol Chem 10:8692–8700. doi:10.1039/
C2OB26531A
24. Lee CW, Hong DH, Han SB, Jong SH, Kim HC, Fine RL,
Lee SH, Kim HM (2002) A novel stereo-selective sulfony-
lurea, 1-[1-(4-aminobenzoyl)-2,3-dihydro-1H-indol-6-sulfonyl]-
4-phenyl-imidazolidin-2-one, has antitumor efficacy in vitro and
in vivo tumor models. Biochem Pharmacol 64:473–480. doi:10.
1016/S0006-2952(02)01105-X
25. Zhang ZJ, Tian J, Wang LT, Wang MJ, Nan X, Yang L, Liu YQ,
Morris-Natschke SL, Lee KH (2014) Design, synthesis and cyto-
toxic activity of novel sulfonylurea derivatives of podophyllotoxin.
Bioorg Med Chem 22:204–210. doi:10.1016/j.bmc.2013.11.035
26. El-Deeb IM, Bayoumi SM, El-Sherbeny MA, Abdel-Aziz AAM
(2010) Synthesis and antitumor evaluation of novel cyclic arylsul-
fonylureas: ADME-T and pharmacophore prediction. Eur J Med
Chem 45:2516–2530. doi:10.1016/j.ejmech.2010.02.038
27. Zhang H, Zhang Y, Wu G, Zhou J, Huang W, Hu X (2009) Synthesis
and biological evaluation of sulfonylurea and thiourea derivatives
substituted with benzene sulfonamide groups as potential hypo-
glycemic agents. Bioorg Med Chem Lett 19:1740–1744. doi:10.
1016/j.bmcl.2009.01.082
28. Ceras J, Cirauqui N, Pérez-Silanes S, Aldana I, Monge A, Galiano
S (2012) Novel sulfonylurea derivatives as H3 receptor antagonists.
Preliminary SAR studies. EurJ Med Chem 52:1–13. doi:10.1016/
j.ejmech.2012.02.049
16. Alqasoumi SI, Al-Taweel AM, Alafeefy AM, Noaman E, Ghorab
MM (2010) Novel quinolines and pyrimido[4,5-b]quinolines bear-
ing biologically active sulfonamide moiety as a new class of
antitumor agents. Eur J Med Chem 45:738–744. doi:10.1016/j.
ejmech.2009.11.021
17. Alqasoumi SI, Al-Taweel AM, Alafeefy AM, GhorabMM Noaman
E (2010) Discovering some novel tetrahydroquinoline derivatives
bearing the biologically active sulfonamide moiety as a new class
of antitumor agents. Eur J Med Chem 45:1849–1853. doi:10.1016/
j.ejmech.2010.01.022
18. Compain G, Martin-Mingot A, Maresca A, ThibaudeauS Supu-
ran CT (2013) Superacid synthesis of halogen containing N-
substituted-4-aminobenzene sulfonamides: new selective tumor-
associated carbonic anhydrase inhibitors. Bioorg Med Chem
21:1555–1563. doi:10.106/j.bmc.2012.05.037
29. León C, Rodrigues J, Gamboa de Domínguez N, Charris J, Gut J,
Rosenthal PJ, Domínguez JN (2007) Synthesis and evaluation of
sulfonylurea derivatives as novel antimalarials. Eur J Med Chem
42:735–742. doi:10.1016/j.ejmech.2007.01.001
30. Boufas W, Dupont N, Berredjem M, Berrezag K, Becheker I,
Berredjem H, Aouf NE (2014) Synthesis and antibacterial activity
of sulfonamides. SAR and DFT studies. J Mol Struct 1074:180–
185. doi:10.1016/j.molstruc.2014.05.066
31. BoucharebF, BoufasW, CheloufiH, BerredjemM, AoufNE(2014)
An efficient method for the synthesis of novel N-acylsulfonamides
using tin (IV) chloride as catalysts. Phosphorus Sulfur Silicon Relat
Elem 189:587–595. doi:10.1080/10426507.2013.843000
32. Berredjem M, Bouchareb F, Ait Kaki S, Dekhil M, Aouf NE (2013)
Synthesis and antibacterial activity of novel N-acylsulfonamides.
Arabian J Chem. doi:10.1016/j.arabjc.2013.01.016
19. Liu F, Martin-Mingot A, Lecornué F, Jouannetaud MP, Maresca
A, Thibaudeau S, Supuran CT (2011) Carbonic anhydrases
inhibitory effects of new benzenesulfonamides synthesized by
using superacid chemistry. J Enzyme Inhib Med Chem 27:886–
891. doi:10.3109/14756366.2011.638921
33. CheloufiH, BerredjemM, BoufasW, BoucharebF, AoufNE(2014)
Efficient synthesis, characterization, and antibacterial activity of
novel N-acylsulfonamides and sulfonylureas. Phosphorus Sulfur
Silicon Relat Elem 189:1396–1404. doi:10.1080/10426507.2013.
865125
20. Mohamadi F, Spees MM, Grindey GB (1992) Antineoplastic
bicyclic sulfonylureas. J Med Chem 35:3012–3016. doi:10.1021/
jm00094a013
21. Medina JC, Shan B, Beckmann H, Farrell RP, Clark DL, Learned
RM, Roche D, Li A, Baichwal V, Case C, Baeuerle PA, Rosen T,
Jaen JC (1998) Novel antineoplastic agents with efficacy against
multidrug resistant tumor cells. Bioorg Med Chem Lett 8:2653–
2656. doi:10.1016/S0960-894X(01)80735-6
22. Mastrolorenzo A, Scozzafava A, Supuran CT (2000) 4-Toluene
sulfonylureido derivatives of amines, amino acids and dipeptides:
a novel class of potential antitumor agents. Eur J Pharm Sci 11:325–
332. doi:10.1016/S0928-0987(00)00122-6
34. Barbey C, Bouasla R, Berredjem M, Dupond N, Retailleau P,
Aouf NE, Lecouvey M (2012) Synthesis and structural study of
new substituted chiral sulfamoyl oxazolidin-2-ones. Tetrahedron
68:9125–9130. doi:10.1016/j.tet.2012.08.001
35. Bouasla R, Berredjem M, Aouf NE, Barbey C (2008) 1,2,3,4-
Tetra-hydro-isoquinoline-2-sulfonamide. Acta Crystallogr Sect E
64:o432. doi:10.1107/S1600536807068158
36. Cheraiet Z, Hessainia S, Ouarna S, Berredjem M, Aouf NE
(2013) A simple and eco-sustainable method for the O-Boc
protection/deprotection of various phenolic structures under water-
mediated/catalyst-free conditions. Green Chem Lett Rev 6:211–
216. doi:10.1080/17518253.2012.738371
23. Rostom SAF (2006) Synthesis and in vitro antitumor evaluation
of some indeno[1,2-c]pyrazol(in)es substituted with sulfonamide,
sulfonylurea(-thiourea) pharmacophores, and some derived thia-
zole ring systems. Bioorg Med Chem 14:6475–6485. doi:10.1016/
j.bmc.2006.06.020
123