Synthesis, anticancer and radiosensitizing evaluation of some novel sulfonamide derivatives

Article abstract of DOI:10.1016/j.ejmech.2015.01.036

In this study, novel series of sulfonamide derivatives were synthesized starting from 2-cyanoacetyl)hydrazono)ethyl)phenyl)benzenesulfonamide 4a and 2-cyanoacetyl)hydrazono)ethyl)phenyl)-4-methylbenzenesulfonamide 4b. Different biologically active moietie

Full text of DOI:10.1016/j.ejmech.2015.01.036

European Journal of Medicinal Chemistry 92 (2015) 682e692
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis, anticancer and radiosensitizing evaluation of some novel
sulfonamide derivatives
, c,
Mostafa M. Ghorab a *, Fatma A. Ragab b, Helmy I. Heiba a, Marwa G. El-Gazzar a,
Sally S. Zahran a
a Department of Drug Radiation Research, National Center for Radiation Research and Technology, Nasr City, Cairo, Egypt
b Department of Pharmaceutical and Medicinal Chemistry, Faculty of Pharmacy, Cairo University, Egypt
c Department of Pharmacognosy, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia
a r t i c l e i n f o
a b s t r a c t
Article history:
In this study, novel series of sulfonamide derivatives were synthesized starting from 2-cyanoacetyl)
Received 5 November 2014
Received in revised form
2 January 2015
Accepted 19 January 2015
Available online 20 January 2015
hydrazono)ethyl)phenyl)benzenesulfonamide
4a
and
2-cyanoacetyl)hydrazono)ethyl)phenyl)-4-
methylbenzenesulfonamide 4b. Different biologically active moieties as pyrazol, thiophene, pyridine
and pyrimidines were introduced in order to investigate their in-vitro anticancer activity, in addition to a
novel series of sulfonamide chalcones were synthesized from the reported 4-acetyl-N-(P-tolyl) benze-
nesulfonamide 3b. The newly synthesized sulfonamide derivatives were characterized by FT-IR, 1H NMR,
13C NMR, mass spectroscopy and elemental analyses and were tested for their in-vitro anticancer activity
against human tumor liver cell line (HEPG-2). The most potent compounds in this study were com-
pounds 4a, 4b, 5a, 6a, 6b, 8, 9, 11, 13, 18 and 19 which showed higher activity than doxorubicin with IC50
Keywords:
Sulfonamide
Anticancer
ranging from 11.0 to 31.8
their ability to enhance the cell killing effect of
m
M. Additionally, eight compounds among the most potent were evaluated for
-radiation.
© 2015 Elsevier Masson SAS. All rights reserved.
Radiosensitizers
g
1. Introduction
[21e25]. Furthermore, particular interest has been focused on the
anticancer activity of chalcones which represent an important
group of the polyphenolic family [26]. This family possesses an
interesting spectrum of biological activities, including anti-
micriobial [27,28], anti-inflammatory [29], immunosuppressive
[30] and anticancer activity [31,32]. Compounds of this family have
been shown to interfere with each step of carcinogenesis, including
initiation, promotion and progression [33]. Moreover, numerous
chalcones appear to show activity against cancer cells, suggesting
that these molecules or their derivatives may be considered as
potential anticancer drugs [34]. In the design of new drugs, the
development of hybrid molecules through the combination of
different pharmacophores may lead to compounds with interesting
biological profiles. In recent years, combination chemotherapy with
agents possessing different mechanisms of action is one of the
methods, which are, being adopted to treat cancer [35,36].
Following this approach and as a part of an ongoing effort to find
alternate chemotherapeutic agents for hepatocellular carcinoma
[11,12,37,38], we herein, report the design and synthesis of novel
sulfonamide derivatives bearing a biologically active pyridine,
thiophene, benzothiophene and chalcone moieties for evaluation
as anticancer on liver human tumor cell lines (HEPG2) and as
The hepatocellular carcinoma (HCC) is the most frequent his-
tological type of primary liver carcinoma and is one of the cancer
types of highest incidence worldwide [1]. The majority of patients
diagnosed with HCC have low recovery rates, and conventional and
modified therapies currently available are rarely beneficial [2]. The
human liver cancer cell line HepG2, established in 1979, is the best
characterized and most frequently used cell line to predict overall
hepatotoxicity [3]. On the other hand, sulfonamides possess many
types of biological activities [4,5]. Also, from the Literature survey it
was found that aryl/heteroaryl sulfonamides may act as antitumor
agents through several mechanisms [6]. Previously we studied
about synthesis and biological activity of some sulfonamide con-
taining different biologically active moieties and we observed many
compounds have promising anticancer activity [7e20]. In addition,
many hetrocyclic compounds incorporating pyridine, thiophene or
benzothiophene moieties showed potential anticancer activities
* Corresponding author. Department of Pharmacognosy, College of Pharmacy,
King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia.
E-mail address: mmsghorab@yahoo.com (M.M. Ghorab).
http://dx.doi.org/10.1016/j.ejmech.2015.01.036
0223-5234/© 2015 Elsevier Masson SAS. All rights reserved.

M.M. Ghorab et al. / European Journal of Medicinal Chemistry 92 (2015) 682e692
683
radiosensitizing agents.
target compounds 5e13 involves reaction of 4a, b with malononi-
trile in dioxane to give (Z)-4-(1-((4,6-diamino-3-cyano-2-
oxopyridin-1(2H)-yl)imino)ethyl)-N-p-substituted-phenyl-benze-
nesulfonamide (5a,b) (Scheme 1). IR spectrum of compound 5a
revealed characteristic strong intensity bands at 3338 and
3210 cmꢀ1 for the introduced two NH2 groups. 1H NMR spectrum
displayed up-field singlet at 4.5 ppm assigned for the CH group and
downfield shifted singlet appearing at 8.5 ppm, due to addition of
NH2 group. 13C NMR exhibited new up-field signal at 83.1 ppm for
CH pyridine,148.4 ppm for (NeCeNH2) and 177.9 ppm for (CeNH2).
IR spectrum of compound 5b revealed characteristic strong bands
at 3389 and 3315 cmꢀ1 for the two of NH2 groups. 1H NMR spec-
trum displayed up-field singlet at 4.1 ppm for the CH group and
downfield shifted singlet appearing at 10.7 ppm, due to addition of
NH2 group. 13C NMR exhibited new up-field signal at 83.1 ppm for
CH pyridine,143.6 ppm for (NeCeNH2) and 177.9 ppm for (CeNH2).
Alternatively, malononitrile was reacted with compounds 4a, b in
the presence of elemental sulfur and in absolute ethanol containing
3 drops of triethyl amine to yield the corresponding thiophene
derivatives 6a, b (Scheme 1) this reaction goes in parallel to the
reported Gewald's thiophene synthesis [41] and their spectroscopic
data were consistent to their chemical structures. The corre-
sponding N-(4-((Z)-1-(2-((E)-2-cyano-3-(dimethylamino) acryloyl)
hydrazono) ethyl) phenyl)-4-methylbenzenesulfonamide 7 was
obtained through the addition of DMF-DMA on 4b in xylene
(Scheme 2). IR spectrum revealed bands at 2929, 2830 cmꢀ1 for
CH_aliphatic. 1H NMR spectrum displayed up-field singlet at
3.3 ppm for the addition of two group of CH3 and downfield shifted
singlet appearing at 7.5 ppm due to introduced CH group. 13C NMR
exhibited new up-field signal at 40.3 ppm for introduced two
methyl group, 98.8 ppm for CeCN and down-field signal at
156 ppm for addition of CH group. Compound (Z)-2-cyano-3-(2-(1-
(4-(4-methylphenylsulfonamido) phenyl) ethylidene) hydrazinyl)-
3-oxopropanedithioic acid 8 was obtained via the reaction of CS2
with 4b in DMF containing KOH (Scheme 2), its IR spectrum
revealed band at 1300 cmꢀ1 for C]S group. 1H NMR spectrum
2. Results and discussion
2.1. Chemistry
The synthetic strategies utilized for the synthesis of the target
compounds are outlined in (Schemes 1e3) which could assemble
entirely the desired sulfonamide derivatives (4a,b-26) from the
strategic starting materials N-(4-acetyl-N-(P-substituted) phenyl
benzenesulfonamide (3a,b) [39,40]. Hence, to develop a more
widely applicable approach for the synthesis of the target com-
pounds we chose to evaluate two synthetic strategies based on
substitution of the acetyl group, the first pathway is the reaction of
the nitrogen nucleophile 2-cyanoacetohydrazide with compounds
3a,b to yield (E)-N-(4-(1-(2-(2-cyanoacetyl) hydrazono) ethyl)-N-
(P- substituted) phenyl benzenesulfonamide 4a,b in good yield
(Scheme 1). The structural assignments to synthesized compounds
were based on their physico-chemical characteristics and spectro-
scopic (IR, 1H NMR, 13C NMR, and mass spectral data) investigations.
IR spectrum of compound 4a revealed characteristic strong in-
tensity bands at 3340, 3219 and 2259 cmꢀ1 for the introduced (2NH
and C^N), respectively confirming the formation of cyanoacetyl
hydrazono derivative. 1H NMR spectrum displayed up-field singlet
at 3.3 ppm for the introduced CH2 group and downfield shifted
singlet appearing at 10.9 ppm due to addition of NH group. 13C NMR
exhibited new up-field signal at 27.3 ppm for CH2 and new signal at
125.7 ppm assigned to C^N. IR spectrum of compound 4b revealed
characteristic strong intensity bands at 3438, 3245 and 2259 cmꢀ1
for the introduced (2NH and C^N), respectively confirming the
formation cyanoacetyl hydrazono derivative. 1H NMR spectrum
displayed up-field singlet at 3.6 ppm for the introduced CH2 group
and downfield shifted singlet appearing at 10.9 ppm, due to addi-
tion of NH group. 13C NMR exhibited new up-field signal at
38.6 ppm for CH2 and new signal at 126.6 ppm ascribed to C^N. A
subsequent ring-closure approach effort for the synthesis of the
Scheme 1. Synthetic pathways for compounds 3a,b-6a,b.

684
M.M. Ghorab et al. / European Journal of Medicinal Chemistry 92 (2015) 682e692
Scheme 2. Synthetic pathways for compounds 7e13.
displayed up-field singlet at 1.5 ppm for SH group and singlet at
3.5 ppm for the addition of CH group. 13C NMR exhibited new up-
field signal at 61.9 ppm for CeC]N and down-field signal at
229.7 ppm for addition of C]S group. The reaction of compound 4b
with cyclohexanone and elemental sulfur yielded the correspond-
down-field doublet at 7.5 ppm, respectively assigned for CHCO
group, and shows doublet of CHCH group at 8.06 ppm. 1H NMR
spectrum of compound 15 show up-field singlet at 2.0 ppm of CH3
group. 13C NMR of compounds 14e19 exhibited new signals at the
range of 121.3 ppm, for CHCO group together with signals at the
range of 145.1 ppm for CH]CH group. Our interest in this area was
to design and synthesize sulfonamide chalcones for structur-
eeactivity relationship (SAR) studies in anticancer activities thus,
the treatment of chalcone 18 with hydroxylamine hydrochloride in
absolute ethanol containing KOH afforded the oxime derivative 20
[45,46] (Scheme 3). IR spectrum revealed broad band at 3451 cmꢀ1
of OH group. 1H NMR spectrum displayed new up-field singlet at
2.3 ppm of OH group. 13C NMR exhibited new down-field-signal at
164.8 ppm due to addition of new C]N group. Compound 18 upon
reacting with different hydrazine derivatives in basic conditions
adopted reaction mechanism involving formation of intermediate
(non-isolable) hydrazones and subsequent addition of NH on the
carbonecarbon double bond of the propenone moiety. Similar
formation of pyrazole derivative in literature showed hydrazine's
ing
(Z)-N-(4-(1-(2-(3-amino-4,5,6,7-tetrahydrobenzo[b]thio-
phene-2-carbonyl) hydrazono) ethyl) phenyl)-4-methyl benzene-
sulfonamide 9 the reaction proceeded according to similar re-
ported reactions [42] (Scheme 2). The dicyanopyridinone de-
rivatives 11 and 13 were obtained through treatment of compound
4b with aromatic benzylidine malononitriles 10 and 12 in dioxane
in the presence of piperidine as a basic catalyst (Scheme 2). Their IR
spectra confirmed the assigned structures by the presence of sig-
nificant band for NH2 and an additional band for CN at their
specified regions. 1H NMR spectra displayed disappearence of
singlet at 3.6 ppm of CH2 group which was involved in the cycli-
zation process and a new signal assigned for NH2 group appeared at
10.4 ppm which was exchangeable with D2O. 13C NMR for com-
pounds 11 and 13 exhibited new up-field signal at 71.7, 76.5 ppm
respectively for CeC]N and signals at 116.18 and 115.8 for CN
group.
attack preferentially on the carbonyl group of a,b-unsaturated ke-
tones, rather than the double bond confirms the formation of
compounds 21e24 [45,46] (Scheme 3). The formation of pyrazole
was confirmed through the spectroscopic data by the disappear-
ance of carbonyl group and the aliphatic bands in their IR spectra
and the appearance of pyrazole protons in the range of 7.1e7.8 in
their 1H NMR spectra. 13C NMR of compounds 21e24 exhibited new
signal of CH-pyrazol at 99.7, 106.7, 103.5 and 104.5 ppm, respec-
tively, signal of C]CeN pyrazol at 147.1, 144.5, 144.8 and
144.3 ppm, respectively and new signal of C]N at 147.7, 153.4,
152.3 and 152.3 ppm, respectively. Compounds 23, 24 showed new
The second pathway adopted for the synthesis of compounds
14e19 depends on the synthesis of novel sulfonamide derivatives
bearing chalcone moieties which are considered an important class
of anticancer agents [43]. The chalcones 14e19 were prepared by
the ClaiseneSchmidt condensation between acetophenone deriv-
ative 3b and aromatic aldehydes in the presence of potassium hy-
droxide in ethanol [44] (Scheme 3). IR spectra of compounds 14e19
showed bands of C]O group at ranges of 1647e1680 cmꢀ1
respectively. 1H NMR spectra of compounds 14e19 displayed
,

M.M. Ghorab et al. / European Journal of Medicinal Chemistry 92 (2015) 682e692
685
Scheme 3. Synthetic pathways for compounds 14e26.
signals for C]S at 176.8 and 174.9 ppm, respectively. Owing to the
2.2. In-vitro anticancer evaluation against human tumor liver
cancer (HEPG2)
significant features of dihydropyrimidine derivatives a number of
protocols for their synthesis have been developed, herein, reaction
of compound 18 with urea and thiourea in ethanol in presence of
KOH gave 25 and 26 following reported method [45,46]. IR of
compound 25 showed band of C]O at 1669 cmꢀ1. While, IR of
compound 26 showed band C]S at 1300 cmꢀ1. 1H NMR spectrum
of compounds 25 and 26 showed singlet for CH at 6.0, 6.7 ppm
respectively, while singlet of NH appeared at 8.5, 13.7 ppm
respectively. 13C NMR of compounds 25 and 26 exhibited new
signal at 106.6, 104.3 ppm of CH group respectively. 13C NMR of
compound 25 showed signal at 156.3 ppm of C]O group, while
spectrum of compound 26 show signal at 180.4 of C]S. Further, the
mass spectra of all the synthesized compounds are in conformity
with the assigned structures.
From the results in Table 1, it was found that compounds 4a, 4b,
5a, 6a, 6b, 8, 9,11,13,18 and 19 were the most potent compounds in
the study with IC50 ranges 11.0e31.8
cytotoxic activities when compared with control and the reference
drug doxorubicin (IC50 ¼ 36.3 M). Compounds 3a, 3b, 5b, 7, 16 and
26 showed equipotent activities than that of the reference drug,
ranging from 35.3 to 39.3 M.
mM and exhibited higher
m
m
A closure look into the structure activity relationship indicates
that the acetophenone derivatives 3a, b showed equipotent activ-
ities as the reference drug (IC50 ¼ 35.3 and 35.8
improved to be 28.0 and 26.0 M in compound 4a, b due to addition
of cyanoacetohydrazone moiety. The addition of pyridinone moiety
mM) which was
m

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M.M. Ghorab et al. / European Journal of Medicinal Chemistry 92 (2015) 682e692
Table 1
In-vitro anticancer screening of the synthesized compounds against HEPG2.
Comp. No.
Control(DMSO)
Concentration (
m
M)
IC50 (mM)
5
12.5
25
50
Surviving fraction (mean SE)a
0.891 0.085
3a
3b
4a
4b
5a
5b
6a
6b
7
8
9
11
13
14
15
16
17
18
19
20
21
22
23
24
1.438 0.026
1.532 0.039
1.373 0.051
1.430 0.026
1.423 0.026
1.514 0.039
1.513 0.039
1.409 0.025
1.738 0.065
1.337 0.049
1.398 0.052
1.543 0.057
1.267 0.047
2.403 0.089
2.160 0.080
2.518 0.093
2.395 0.088
1.662 0.061
2.362 0.087
2.493 0.092
2.365 0.087
2.039 0.075
2.165 0.079
1.248 0.046
2.234 0.082
1.535 0.056
0.719 0.028
0.726 0.068
0.778 0.056
0.530 0.058
0.556 0.009
0.761 0.003
0.456 0.052
0.383 0.026
0.877 0.045
0.229 0.016
0.528 0.026
0.696 0.042
0.635 0.047
2.032 0.103
1.616 0.149
0.497 0.068
1.478 0.027
0.348 0.043
0.505 0.054
1.769 0.231
1.197 0.244
1.841 0.129
1.759 0.058
1.189 0.012
0.950 0.057
0.805 0.015
0.546 0.020
0.464 0.067
0.547 0.247
0.361 0.023
0.349 0.042
0.312 0.044
0.577 0.051
0.239 0.035
0.390 0.017
0.546 0.033
0.291 0.049
0.331 0.052
0.363 0.042
0.331 0.047
1.017 0.125
0.781 0.019
0.383 0.069
0.591 0.082
0.392 0.072
0.430 0.037
1.351 0.267
0.522 0.012
0.923 0.124
1.078 0.189
0.507 0.100
0.732 0.081
0.486 0.034
0.461 0.010
0.417 0.030
0.367 0.116
0.242 0.025
0.293 0.026
0.321 0.023
0.385 0.030
0.274 0.047
0.323 0.029
0.378 0.059
0.414 0.055
0.277 0.012
0.375 0.044
0.408 0.042
0.529 0.019
0.489 0.037
0.551 0.079
0.402 0.021
0.495 0.054
0.445 0.008
0.632 0.076
0.508 0.039
0.536 0.114
0.500 0.061
0.530 0.066
0.481 0.078
0.398 0.016
0.494 0.030
35.3
35.8
28.0
26.0
25.5
37.0
23.3
21.8
37.8
11.0
25.5
31.8
31.5
NA
49.8
39.3
43.3
26.0
30.3
NA
1.048 0.073
0.881 0.054
0.837 0.027
0.748 0.049
0.937 0.017
0.869 0.078
0.571 0.034
1.141 0.109
0.344 0.001
0.868 0.024
0.984 0.058
0.951 0.036
2.385 0.105
1.931 0.270
1.461 0.167
2.16 0.189
0.476 0.085
0.584 0.033
1.723 0.209
1.772 0.027
2.085 0.081
1.907 0.054
1.766 0.199
1.619 0.164
1.066 0.026
0.721 0.020
44.8
NA
NA
45.3
46.3
36.5
36.3
25
26
Doxorubicin
a
Each value is the mean of three values standard Error; NA: no activity.
was proved to be successful in case of compound 5a which showed
in vitro anticancer activity with IC50 value of 25.5 mM, when the
an increase in the activity (IC50 ¼ 25.5
m
m
M). Oppositely, the activity
M. The efficiency of intro-
cells were subjected to different concentrations of the compound
alone. While when the cells were subjected to the same concen-
trations of compound 5a, and irradiated with a single dose of ɤ-
radiation at a dose level of 8 Gy, the IC50 value was synergistically
of compound 5b decreased to be 37.0
ducing thiophene ring was proved in compounds 6a, b whose ac-
M) in
tivities were interestingly improved (IC50 ¼ 23.3 and 21.8
m
comformity with the well-documented anticancer activity of
thiophene derivatives [47]. On the other hand introduction of
dimethyl amino group through the reaction of 4b with DMF-DMA
decreased to 11.5
23.3 M when used alone. The IC50 value was decreased to 18.3
when the cells were treated with compound 6a in combination
with ɤ-radiation. Also, compound 6b showed IC50 value of 21.8
when tested alone. The IC50 value was decreased to 5.8 M, when
mM. Similarly, compound 6a showed IC50 value of
m
mM,
yielded compound
(IC50 ¼ 37.8 M). While, the addition of bifunctional CS2 afforded
compound 8 which is the most potent candidate in this screening
(IC50 ¼ 11.0 M). Similarly, introduction of tetrahydrobenzo thio-
phene moiety resulted in high activity compound
(IC50 ¼ 25.5 M). Concerning the dicyanopyridinone derivatives 11
and 13 they both showed lower activities (IC50 ¼ 31.8 and 31.5 M)
compared to the corresponding starting material 4b
M) but still they are more potent than doxorubicin.
7
which showed relatively low activity
mM
m
m
the cells were treated with compound 6b in combination with ɤ-
radiation. Similar findings for compounds 8, 9, 13, 18 and 19 which
m
9
showed IC50 values of 11.0, 25.5, 31.5, 26.0 and 30.3
tively, while, their IC50 values decreased synergistically with radi-
ation to become 5.0, 20.0, 23.5, 20.5 and 26.8 M. The results
mM, respec-
m
m
m
proved the ability of the synthesized compounds to sensitize cancer
cells to the lethal effects of ionizing radiation in order to decrease
the dose of the drug and decrease its toxicity. The change in IC50
(IC50 ¼ 26.0
m
The synthesis of chalcone derivatives 14e19 indicated that the
most potent among the six new compounds was the p-nitrophenyl
(mM) for compounds 5a, 6a, 6b, 8, 9, 13, 18 and 19 against HEPG2
derivative 18 (IC50 ¼ 26.0
m
M) and this finding encouraged us to
before and after radiation is illustrated in Fig. 1.
synthesize more derivatives by introducing oxime, pyrazole and
pyrimidine moieties in compounds 20e26, respectively, aiming to
obtain more potent anticancer agents but, unfortunately, the
resulting compounds were with low activities except for compound
3. Conclusion
In summary, we had synthesized a novel series of sulfonamide
derivatives. Most of the prepared compounds revealed potential
anticancer activity against human liver cancer cell line. Compounds
4a, 4b, 5a, 6a, 6b, 8, 9, 11, 13, 18 and 19 were found to be more
potent than doxorubicin. From the structureeactivity relationships,
we may conclude that the introduction of carbon disulphide is
associated with enhanced anticancer activity and gave the most
26 (IC50 ¼ 36.5
mM) and these results proved the importance of a,b-
unsaturated ketone system in chalcones as anticancer pharmaco-
phore without any substitution [48,49].
2.3. Radiosensitizing evaluation
The ability of the most active compounds 5a, 6a, 6b, 8, 9, 13, 18
and 19 to enhance the cell killing effect of ɤ-irradiation was studied.
From the results obtained in Table 2, compound 5a showed an
potent compound in this study (IC50 ¼ 11.0
sulfonamide with chalcone moiety in compound 18 increased the
activity (IC50 ¼ 26.0 M), but further substitution on
mM). Combination of
m
a, b-

M.M. Ghorab et al. / European Journal of Medicinal Chemistry 92 (2015) 682e692
687
Table 2
In-vitro anticancer screening of compounds 5a, 6a, 6b, 8, 9, 13, 18 and 19 against human liver cancer cell line (HEPG2) after exposure to ɤ-radiation.
Comp. No.
Control (radiation)
Concentration (
5
m
M)
IC50 (mM)
12.5
25
50
Surviving fraction (mean SE)a
5a
6a
6b
8
1.116 0.041
1.011 0.037
1.232 0.045
1.123 0.040
0.917 0.033
1.032 0.037
1.215 0.044
1.187 0.042
0.395 0.095
0.526 0.015
0.291 0.037
0.245 0.019
0.521 0.031
0.439 0.056
0.558 0.040
0.591 0.051
0.194 0.194
0.386 0.090
0.227 0.010
0.206 0.007
0.524 0.017
0.603 0.262
0.451 0.122
0.471 0.025
0.299 0.021
0.253 0.004
0.105 0.019
0.189 0.043
0.329 0.024
0.345 0.049
0.368 0.039
0.394 0.079
0.340 0.043
0.322 0.016
0.174 0.006
0.158 0.008
0.229 0.010
0.352 0.027
0.249 0.143
0.408 0.033
11.5
18.3
5.80
5.00
9
20.0
13
18
19
23.5
20.5
26.8
Each value is the mean of three values Standard Error; Significant difference from control group at P < 0.001.
4.1.1. (E)-N-(4-(1-(2-(2-cyanoacetyl)hydrazono)ethyl)phenyl)
benzenesulfonamide (4a)
A mixture of 2ecyanoacetohydrazide (2.76 g, 0.01 mol) and 4-
acetyl-N-phenyl benzenesulfonamide 3a [31] (1 g, 0.01 mol) in
dioxane (10 mL), was refluxed for 3 h, then left to cool. The solid
product formed upon pouring onto ice/water was collected by
filtration and recrystallized from ethanol to give 4a. Yield, 69%; m.p.
120e122 ꢁC. IR (KBr, cmꢀ1): 3340, 3219 (2NH), 3058 (CH arom.),
2925, 2830 (CH aliph.), 2259 (C^N),1667 (C]O),1596 (C]N),1347,
1162 (SO2). H1NMR (DMSO-d6): 2.1 [s, 3H, CH3], 3.3 [s, 2H, CH2],
6.6e7.2 [m, 5H, AreH], 8.0 [s, 1H, SO2NH exchangeable with D2O],
7.8, 7.7 [2d, 4H, AreH AB system, J ¼ 6.8 Hz], 10.9 [s, 1H, NH
exchangeable with D2O]. C13NMR (DMSO-d6): 26.3, 27.3 (CH2),
117.9, 122.4, 125.7 (C^N), 126.6, 129.5 (2C), 129.7, 139.2, 142.1 (2C),
196.3 (C]O). MS m/z (%): 356 (Mþ) (1.4), 52.2 (100). Anal. Calcd. For
C17H16N4O3S (356): C, 57.29; H, 4.52; N, 15.72. Found: C, 57.53; H,
Fig. 1. Change in IC50 (mM) for compounds 5a, 6a, 6b, 8, 9, 13, 18 and 19 against HEPG2
4.32; N, 4.32.
before and after radiation.
4.1.2. (E)-N-(4-(1-(2-(2-cyanoacetyl)hydrazono)ethyl)phenyl)-4-
methylbenzenesulfonamide (4b)
unsaturated ketone system led to less active compounds. This study
may provide valuable information for further of more potent
anticancer agents. Moreover, the most active compounds showed
interesting radiosensitizing activity when evaluated for their in-
A mixture of 2ecyanoacetohydrazide (2 g, 0.007 mol) and in, 4-
acetyl-N-(P- tolyl)benzenesulfonamide 3b [32] (0.74 g, 0.007 mol)
in dioxane (10 mL) was refluxed for 3 h, then left to cool. The solid
product formed upon pouring onto ice/water was collected by
filtration and recrystallized from ethanol to give 4b. Yield, 72.2%;
m.p. 200e202 ꢁC. IR (KBr, cmꢀ1): 3438, 3245 (2NH), 3058 (CH
arom.), 2927, 2830 (CH aliph.), 2259 (C^N), 1680 (C]O), 1593 (C]
N), 1349, 1156 (SO2). H1NMR (DMSO-d6): 2.1 [s, 3H, CH3], 2.5 [s, 3H,
CH3 tolyl], 3.6 [s, 2H, CH2], 7.2, 7.3 [2d, 4H, AreH AB system, J ¼ 6.8
Hz], 8.0 [s, 1H, SO2NH exchangeable with D2O], 7.7, 7.8 [2d, 4H,
AreH AB system, J ¼ 7.1 Hz], 10.9 [s,1H,NH exchangeable with D2O].
C13NMR (DMSO-d6): 20.8 (CH3 tolyl), 26.2, 38.6 (CH2), 117.8, 126.6
(C^N), 127.1, 129.7, 129.8, 131.8, 136.4, 142.3, 143.5 (2C), 196.3 (C]
O). MS m/z (%): 370 (Mþ) (1.02), 90 (100). Anal. Calcd. For
vitro anticancer activity in combination with g-irradiation.
4. Experimental
4.1. Chemistry
Melting points were uncorrected and were taken in an open
capillary tube on a Stuart melting point apparatus (Stuart Scientific,
Redhill, UK). The IR spectra of the compounds were recorded on
ABB Bomem FT-IR spectrometer MB 104 with KBr pellets. 1H NMR
and 13C NMR spectra were recorded using a Bruker 300 NMR
spectrometer operating at 400.13 and 100.77 MHz, respectively.
Microanalyses were obtained with Elemental analyses system
GmbH VarioEL V300 element analyzers which were found within
the limit of 0.4% of theoretical values for all the synthesized com-
pounds. The purity of the compounds was checked by thin layer
chromatography (TLC) on pre-coated silica gel plates (Kiesel gel
0.25 mm, 60 G F 254, Merck)). The developing solvent system was
chloroform/methanol (10:3) and the spot were visualized in UV
chamber. IR, 1H NMR, 13C NMR and elemental analysis were
consistent with the assigned structures and performed at the
Microanalytical Laboratories of the Faculty of Science, Cairo
University.
C18H18N4O3S (370): C, 58.36; H, 4.90; N, 15.12. Found: C, 58.70; H,
4.60; N, 15.40.
4.1.3. (Z)-4-(1-((4,6-diamino-3-cyano-2-oxopyridin-1(2H)-yl)
imino)ethyl)-N-phenyl-benzenesulfonamide (5a)
Equimolecular amounts of compound 4a (0.5 g, 0.0013 mol) and
malononitrile (0.06 g, 0.0013 mol) in dioxane (10 mL) containing
triethylamine (5dps) was heated under reflux for 6 h, then left to
cool. The solid product formed upon pouring onto ice/water was
collected by filtration and recrystallized from 1,4-dioxane to give
5a. Yield, 63.8%; m.p. 196e198 ꢁC. IR (KBr, cmꢀ1): 3338, 3210, 3199
(NH2, NH), 3058 (CH arom.), 2925, 2864 (CH aliph.), 2273 (C^N),
1692 (C]O), 1510 (C]N), 1329, 1162 (SO2).
H1NMR (DMSO-d6): 2.1 [s, 3H, CH3], 4.5 [s, 1H, CH-pyridine],
6.6e7.0 [m, 5H, AreH], 8.0 [s, 1H, SO2NH exchangeable with D2O],

688
M.M. Ghorab et al. / European Journal of Medicinal Chemistry 92 (2015) 682e692
7.7, 7.8 [2d, 4H, AreH AB system, J ¼ 7.1 Hz], 8.5 [s, 4H, 2 NH2
exchangeable with D2O]. C13NMR (DMSO-d6): 13.5 (CH3), 69.9
(CeCN), 83.1 (CH-pyridine), 115.8 (C^N), 119.0, 122.4, 127.3, 127.3,
129.2, 129.5, 138.6, 142.0, 148.4 (NeCeNH2), 160.0 (C]O), 165.6,
177.9 (CeNH2). MS m/z (%): 423 (M þ 1) (3.49), 90.70 (100). Anal.
Calcd. For C20H18N6O3S (422): C, 56.86; H, 4.29; N, 19.89. Found: C,
56.6; H, 4.41; N, 19.5.
143.3,143.6,152.0 (CeNH2),165.5 (SeCeNH2),196.3 (C]O). MS m/z
(%): 466 (Mꢀ2) (0.23), 90 (100). Anal. Calcd. For C21H20N6O3S2
(468): C, 53.83; H, 4.30; N, 17.94. Found: C, 53.75; H, 4.52; N, 17.74.
4.1.7. N-(4-((Z)-1-(2-((E)-2-cyano-3-(dimethylamino)acryloyl)
hydrazono)ethyl)phenyl)-4-methylbenzenesulfonamide (7)
A mixture of 4b (0.3 g, 0.0008 mol) and dimethylformamide-
dimethylacetal (DMF-DMA) (0.09 g (0.11 ml), 0.0008 mol), in dry
xylene (15 ml) was refluxed for 3 h and filtered, then the filtrate was
cooled in refrigerator then filtered. The obtained soild was recrys-
tallized from ethanol to give 7. Yield, 26.4%; m.p. 185e187 ꢁC. IR
(KBr, cmꢀ1): 3240 (NH), 3200 (CH arom.), 2929, 2830 (CH aliph.),
2288 (C^N), 1666 (C]O), 1588 (C]N), 1338, 1156 (SO2). H1NMR
(DMSO-d6): 2.1 [s, 6H, CH3], 2.5 [s, 3H, CH3 tolyl], 3.3 [s, 6H, N-
dimethyl], 7.2, 7.3 [2d, 4H, AreH AB system, J ¼ 6.9 Hz], 7.5 [s, 1H,
C]CH], 7.7, 7.8 [2d, 4H, AreH AB system, J ¼ 7.2 Hz], 8.0 [s, 1H,
SO2NH exchangeable with D2O], 10.7 [s, 1H, NH exchangeable with
D2O]. C13NMR (DMSO- d6): 20.8, 26.2, 40.3 (N(CH3)2), 98.8 (CeCN),
117.8, 118.8 (C^N), 127.4, 129.7, 129.8, 131.8, 136.4, 142.3, 143.5,
143.6, 156.0 (CH), 196.3 (C]O). MS m/z (%): 420 (Mꢀ5) (0.05), 92.45
(100). Anal. Calcd. For C21H23N5O3S (425): C, 59.28; H, 5.45; N,
16.46. Found: C, 59.08; H, 5.65; N, 16.26.
4.1.4. (E)-N-(4-(1-((4,6-diamino-3-cyano-2-oxopyridin-1(2H)-yl)
imino)ethyl)phenyl)-4-methylbenzenesulfonamide (5b)
Equimolecular amounts of compound 4b (0.5 g, 0.0013 mol) and
malononitrile (0.06 g, 0.0013 mol) in dioxane (10 mL) containing
triethylamine (5dps) was heated under reflux for 6 h, then left to
cool. The soild product formed upon pouring onto ice/water was
collected by filtration and recrystallized from dioxane to give 5b.
Yield, 44.16%; m.p. 146e148 ꢁC. IR (KBr, cmꢀ1): 3389, 3315, 3219
(NH2, NH), 3058 (CH arom.), 2930, 2830 (CH aliph.), 2196 (C^N),
1661 (C]O), 1590 (C]N), 1349, 1156 (SO2). H1NMR (DMSO-d6): 2.1
[s, 3H, CH3], 2.5 [s, 3H, CH3 tolyl], 4.1 [s,1H, CH pyridine], 7.2, 7.3 [2d,
4H, AreH AB system, J ¼ 6.8 Hz], 7.7, 7.8 [2d, 4H, AreH AB system,
J ¼ 7.2 Hz ], 8.0 [s, 1H, SO2NH exchangeable with D2O], 10.7 [s, 4H,2
NH2 exchangeable with D2O]. C13NMR (DMSO-d6): 20.8, 26.2, 66.2
(CeC^N), 83.1 (C-pyridine), 117.8, 118.8 (C^N), 127.1, 129.6, 129.8,
131.8, 136.4, 142.2, 143.5, 143.6 (NeCeNH2), 143.6, 177.9 (CeNH2),
196.3 (C]O). MS m/z (%): 431 (Mꢀ5) (0.12), 104 (100). Anal. Calcd.
For C21H20N6O3S (436): C, 57.79; H, 4.62; N, 19.25. Found: C, 58; H,
58; N, 19.52.
4.1.8. (Z)-2-Cyano-3-(2-(1-(4-(4-methylphenylsulfonamido)
phenyl)ethylidene)hydrazinyl)-3-oxopropanedithioic acid (8)
Carbon disulfide (0.0061 g, 0.0008 mol) was added gradually to
a
cold solution of the acetohydrazide derivative 4b (0.3 g,
0.0008 mol) in N,N-dimethylformamide (5 mL) containing finely
grounded potassium hydroxide (1.0 g) and the reaction mixture
was left at room temperature for an additional 24 h with stirring.
The reaction mixture was then triturated with cold water and
neutralized with 1N HCl. The resulting precipitated soild was
collected by filtration, washed with water, dried and recrystalized
from dioxane to give 8. Yield, 98.31%; m.P. 146e148 ꢁC. IR (KBr,
cmꢀ1): 3380, 3213 (2 NH), 3058 (CH arom.), 2926, 2830 (CH aliph.),
2288 (C^N), 1664 (C]O), 1590 (C]N), 1338, 1156 (SO2), 1300 (C]
S). H1NMR (DMSO-d6): 1.5 [s, 1H, SH exchangeable with D2O ], 2.1
[s, 3H, CH3], 2.5 [s, 3H, CH3 tolyl], 3.5 [s, 1H, CH], 7.0 [s, 1H, NH
exchangeable with D2O], 7.2, 7.3 [2d, 4H, AreH AB system, J ¼ 6.8
Hz], 7.7, 7.8 [2d, 4H, AreH AB system, J ¼ 7.3 Hz], 8.0 [S, 1H, SO2NH
exchangeable with D2O]. C13NMR (DMSO-d6): 20.8, 26.27, 61.9
(CeC^N), 115.3 (C^N), 117.8, 127.1, 129.6, 129.8, 131.8, 136.4, 140.7,
142.2,143.6,171 (C]O), 229.7 (C]S). MS m/z (%): 444 (Mꢀ2) (3.08),
63.5 (100). Anal. Calcd. For C19H18N4O3S3 (446): C, 51.10; H, 4.06; N,
12.55. Found: C, 51.0; H, 4.31; N, 12.33.
4.1.5. (E)-N-(4-(1-(2-(3,5-diamino-4-cyanothiophene-2-carbonyl)
hydrazono)ethyl)phenyl)-benzenesulfonamide (6a)
To a solution of compound 4a (0.35 g, 0.0013 mol)in absolute
ethanol (10 mL) containing triethylamine (5dps) malononitrile
(0.06 g, 0.0013 mol) together with elemental sulfur (0.032 g,
0.001 mol) were added. The reaction mixture was heated under
reflux for 2 h, then left to cool and poured onto ice/water and the
solid obtained was recrystallized from dioxane to give 6a. Yield,
82.2%; m.p.167e169 ꢁC. IR (KBr, cmꢀ1): 3390, 3250, 3194 (NH2, NH),
3058 (CH arom.), 2928, 2830 (CH aliph.), 2210 (C^N), 1692 (C]O),
1595 (C]N), 1328, 1152 (SO2). H1NMR (DMSO-d6): 2.16 [s, 3H, CH3],
6.6e7.2 [m, 5H, AreH], 6.2, 6.9 [s, 4H, 2NH2 exchangeable with
D2O], 7.0 [s, 1H, NH exchangeable with D2O], 8.0 [s, 1H, SO2NH
exchangeable with D2O], 7.7, 7.8 [2d, 4H, AreH AB system, J ¼ 7.2
Hz]. C13NMR (DMSO-d6): 13.5, 81.1 (CeCN), 117.9 (C^N), 119, 122.4,
127.19, 129.2, 129.3, 129.7, 132.9 (COC), 139.2 (CeS), 142.1, 150.0,
157.8 (CeNH2), 160.8 (SeCeNH2), 169.3 (C]O). MS m/z (%): 454
(Mþ) (0.25), 90 (100). Anal. Calcd. For C20H18N6O3S2 (454): C, 52.85;
H, 3.99; N, 18.49. Found: C, 52.64; H, 4.33; N, 18.58.
4.1.9. (Z)-N-(4-(1-(2-(3-amino-4,5,6,7-tetrahydrobenzo[b]
thiophene-2-carbonyl)hydrazono)-ethyl)phenyl)-4-
4.1.6. (E)-N-(4-(1-(2-(3,5-diamino-4-cyanothiophene-2-carbonyl)
hydrazono)ethyl)phenyl)-4-methylbenzenesulfonamide (6b)
methylbenzenesulfonamide (9)
To a solution mixture of compound 4b (0.5 g, 0,0013 mol), ab-
solute ethanol (10 mL), triethylamine (5dps) and cyclohexanone
(0.13 g, 0.0013 mol), elemental sulfur (0.032 g, 0.0013 mol) was
added. The reaction mixture was heated under reflux for 2 h, then
left to cool and poured onto ice/water. The solid obtained was
recrystallized from dioxane to give 9. Yield, 56.45%; m.p.
110e112 ꢁC. IR (KBr, cmꢀ1): 3390, 3319, 3222 (NH2, NH), 3058 (CH
arom.), 2930, 2830 (CH aliph.), 1660 (C]O), 1580 (C]N), 1349, 1156
(SO2). H1NMR (DMSO-d6): 1.7, 2.7 [m, 8H,CH2 cyclohexane], 2.2 [s,
3H, CH3], 2.5 [s, 3H, CH3 tolyl], 7.2, 7.3 [2d, 4H, AreH AB system,
J ¼ 6.9 Hz], 7.7, 7.8[2d, 4H, AreH AB system, J ¼ 7.7 Hz], 8.0 [s, 1H,
SO2NH exchangeable with D2O], 10.4 [s, 2H, NH2 exchangeable with
D2O], 10.8 [s, 1H, NH exchangeable with D2O]. C13NMR (DMSO-d6):
20.8, 23.4 (C-hexane), 24.9 (C-hexane), 26.7, 117.8, 127.1, 129.6,
129.8, 131.8, 134.5, 135.2 (O]CeCeS), 136.4, 137.1 (CeNH2), 142.2,
143.5, 143.6, 147.4 (CeS), 169.3 (C]O). MS m/z (%): 482 (Mþ) (1.34),
To a solution of compound 4b (0.37 g, 0,0013 mol) in absolute
ethanol (10 ml) containing triethylamine (5dps) malononitrile
(0.06 g, 0.0013 mol) together with elemental sulfur (0.032 g,
0.0013 mol) were added. The reaction mixture was heated under
reflux for 2 h, then left to cool and poured onto ice/water and the
solid obtained was recrystallized from dioxane to give 6b. Yield,
83.3%; m.p. 150e152 ꢁC. IR (KBr, cmꢀ1): 3391, 3311, 3219 (NH2, NH),
3058 (CH arom.), 2928, 2830 (CH aliph.), 2202 (C^N), 1664 (C]O),
1596 (C]N), 1349, 1156 (SO2). H1NMR (DMSO-d6): 2.16 [s, 3H, CH3],
2.5 [s, 3H, CH3 tolyl], 7.2, 7.3[ 2d, 4H, AreH AB system, J ¼ 6.8 Hz],
7.7, 7.8 [2d, 4H, AreH AB system, J ¼ 7.1 Hz], 8.0 [s, 1H, SO2NH
exchangeable with D2O], 10.5 [s, 2H, NH2 exchangeable with D2O],
10.7 [s, 2H, NH2 exchangeable with D2O], 10.9 [s, H, NH exchange-
able with D2O]. C13NMR (DMSO-d6): 20.8, 26.2, 76.5 (CeC^N), 116
(C^N), 117.8, 127.5, 129.6, 129.7, 131.8, 132.9 (CeS), 136.4, 142.26,

M.M. Ghorab et al. / European Journal of Medicinal Chemistry 92 (2015) 682e692
689
185.20 (100). Anal. Calcd. For C24H26N4O3S2 (482): C, 59.73; H, 5.43;
N, 11.61; Found: C, 59.55; H, 5.63; N, 11.58.
(DMSO-d6): 20.8, 116.8, 121.3 (CHCO), 127.9, 128.0, 128.3, 128.5,
128.6, 129.3, 132.0, 135.2, 136.7, 137.6, 143.5, 145.1 (HC]CH), 185.6
(C]O). MS m/z (%): 377(Mþ) (0.28), 92.55(100). Anal. Calcd. For
4.1.10. (Z)-N-(4-(1-((6-amino-3,5-dicyano-4-(4-methoxyphenyl)-
2-oxopyridin-1(2H)-yl)imino)-ethyl)phenyl)-4-
C22H19NO3S (377): C, 70.00; H, 5.07; N, 3.71. Found: C, 70.2; H, 5.27;
N, 3.51.
methylbenzenesulfonamide (11)
Equimolecular mixture of 4b (0.5 g, 0.0013 mol) and 2-(4-
methoxybenzylidene) malononitrile 10 (0.18 g, 0.0013 mol) in
dioxane (5 ml) containing piperidine (0.5 ml), was heated under
reflux for 4 h, then left to cool and poured onto ice/water containing
few drops of HCl and the formed solid product was recrystallized
from 1,4-dioxane to give 11. Yield, 67.6%; m.p. 116e118 ꢁC. IR (KBr,
cmꢀ1): 3390, 3301, 3214 (NH2, NH), 3058 (CH arom.), 2925, 2830
(CH aliph.), 2208 (2C^N), 1678 (C]O), 1580 (C]N), 1349, 1156
(SO2).
4 .1.12 . 2 . ( E ) - N - ( 4 - ( 3 - ( p - t o l y l ) a c r y l o y l ) p h e n y l ) - 4 -
methylbenzenesulfonamide (15). Yield, 63.49%; m.p. 225e227 ꢁC. IR
(KBr, cmꢀ1): 3318 (NH), 3032 (CH arom.), 2920, 2857 (CH aliph.),
1680 (C]O), 1308, 1110 (SO2). H1NMR (DMSO-d6): 2.0 [s, 6H, CH3
tolyl, p-CH3], 7.1, 7.3 [2d, 4H, AreH AB system, J ¼ 6.3 Hz], 7.5 [d, 1H,
CHCO, J ¼ 8.1 Hz], 7.9, 8.0 [ 2d, 4H, AreH AB system, J ¼ 7.1 Hz], 8.0
[s, 1H, SO2NH], 8.06 [d,1H, HC]CH, J ¼ 6.9 Hz], 8.2, 8.3 [2d,4H,
AreH AB system, J ¼ 7.1 Hz]. C13NMR (DMSO-d6): 20.8 (2CH3),118.9,
121.3 (CHCO), 127.9, 128.1, 128.2, 128.3, 129.3, 132.0, 132.2, 136.7,
137.6 (2Ce CH3), 143.5, 145.1 (HC]CH), 185.2 (C]O). MS m/z (%):
390 (Mꢀ1) (0.26), 120.40 (100). Anal. Calcd. For C23H21NO3S (391):
C, 70.56; H, 5.41; N, 3.58. Found: C, 70.36; H, 5.31; N, 3.88.
H1 NMR (DMSO-d6): 2.1 [s, 3H, CH3], 2.5 [s, 3H, CH3 tolyl], 3.8 [s,
3H, OCH3], 7.1, 7.2 [2d, 4H, AreH AB system, J ¼ 6.8 Hz], 7.3, 7.4 [2d,
4H, AreH AB system, J ¼ 7.4 Hz], 7.6, 7.9 [2d, 4H, AreH AB system,
J ¼ 8.1 Hz], 8.0 [ s, 1H, SO2NH exchangeable with D2O], 10.9 [s, 2H,
NH2 exchangeable with D2O]. C13NMR (DMSO-d6): 20.8, 26.2, 55.6
(OCH3), 71.7 (CeCN), 113.1(C-phenyl), 116.1, 116.1 (C^N), 118.1,
127.1, 129.1, 129.1, 130.1 (C-phenyl), 132.1, 136.1, 138.1 (C-phenyl),
142.2,143.1, 148.1 (CeOCH3),159.5 (CeNH2), 165.1 (C]O),169.4. MS
m/z (%): 551 (Mꢀ1) (0.89), 50.55 (100). Anal. Calcd. For
4.1.12.3. (E)-N-(4-(3-(4-fluorophenyl)acryloyl)phenyl)-4-
methylbenzenesulfonamide (16). Yield, 97.6%; m.p. 208e210 ꢁC. IR
(KBr, cmꢀ1): 3377 (NH), 3199 (CH arom.), 2924, 2850 (CH aliph.),
1652 (C]O), 1305, 1125 (SO2). H1NMR (DMSO-d6): 2.0 [s, 3H,
CH3tolyl], 7.1, 7.3 [2d, 4H, AreH AB system, J ¼ 6.3 Hz], 7.5 [d, 1H,
CHCO, J ¼ 6.9 Hz], 7.9, 8.0 [2d,4H, AreH AB system, J ¼ 6.9 Hz ], 8.0
[s, 1H, SO2NH exchangeable with D2O], 8.06 [d,1H, HC]CH, 12 Hz],
8.2, 8.3 [2d,4H, AreH AB system, J ¼ 7.1 Hz]. C13NMR (DMSO-d6):
20.8 (CH3), 117.94, 118.9, 121.3 (CHCO), 126.7, 127.9, 129.3, 129.4,
129.9, 132.0, 136.4, 137.6, 143.6, 145.1 (HC]CH), 162.1 (CeF), 185.23
(C]O). MS m/z (%): 395 (Mþ) (3.09), 65.50 (100). Anal. Calcd. For
C
29H24N6O4S (552): C, 63.03; H, 4.38; N, 15.21; Found: C, 63.33; H,
4.55; N, 15.41.
4.1.11. (Z)-N-(4-(1-((6-amino-3,5-dicyano-4-(4-nitrophenyl)-2-
oxopyridin-1(2H)-yl)imino)-ethyl)phenyl)-4-
methylbenzenesulfonamide (13)
Equimolecular mixture of 4b (0.5 g, 0.0013 mol) and 2-(4-
nitrobenzylidene) malononitrile 12 (0.199 g, 0.0013 mol) in
dioxane (5 ml) containing piperidine (0.5 ml), was heated under
reflux for 4 h, then left to cool and poured onto ice/water containing
few drops of HCL and the formed solid product was recrystallized
from dioxane to give 13. Yield, 45.20%; m.p. 143e145 ꢁC. IR (KBr,
cmꢀ1): 3394, 3344, 3216 (NH2, NH), 3058 (CH arom.), 2925, 2830
(CH aliph.), 2208 (2C^N), 1660 (C]O), 1580 (C]N), 1349, 1156
(SO2). H1 NMR (DMSO-d6): 2.1 [s, 3H, CH3], 2.5 [s, 3H, CH3 tolyl], 7.1,
7.2 [2d, 4H, AreH AB system, J ¼ 6.6 Hz], 7.3, 7.4 [2d, 4H, AreH AB
system, J ¼ 6.9 Hz], 7.6, 7.9 [2d, 4H, AreH AB system, J ¼ 7.1 Hz], 8.5
[s, 1H, SO2NH exchangeable with D2O], 10.4 [s, 2H, NH2 exchange-
able with D2O ]. C13NMR (DMSO-d6): 20.8, 26.2, 76.5 (CeC^N),
115.8 (C^N), 117.8, 123.8 (C- phenyl), 127.6, 129.6, 129.8, 130.1 (C-
phenyl), 131.4, 136.4, 138.6 (C- phenyl), 142.62, 143.59, 143.6, 147.1
(CeNO2), 159.5 (CeNH2), 160 (C]O), 169.4. MS m/z (%): 565 (Mꢀ2)
(o.54), 75 (100). Anal. Calcd. For C28H21N7O5S (567): C, 59.25; H,
3.73; N, 17.27; Found C, 59.55; H, 3.43; N, 17.52.
C22H18FNO3S (395): C, 66.82; H, 4.59; N, 3.54. Found: C, 66.52; H,
4.89; N, 3.34.
4.1.12.4. (E)-N-(4-(3-(4-chlorophenyl)acryloyl)phenyl)-4-
methylbenzenesulfonamide (17). Yield, 98.20%; m.p. >300 ꢁC. IR
(KBr, cmꢀ1): 3364 (NH), 3038 (CH arom.), 2930, 2838 (CH aliph.),
1660 (C]O), 1315, 1134 (SO2), 750 (CeCl). H1NMR (DMSO-d6): 2.0
[s, 3H, CH3 tolyl], 7.1, 7.3 [2d, 4H, AreH AB system, J ¼ 6.3 Hz], 7.5 [d,
1H, CHCO, J ¼ 7.5 Hz], 7.9, 8.0 [2d, 4H, AreH AB system, J ¼ 7.1 Hz],
8.0 [s, 1H, SO2NH exchangeable with D2O], 8.06 [d, 1H, HC]CH,
J ¼ 7.5 Hz], 8.2, 8.3 [2d, 4H, AreH AB system, J ¼ 7.1 Hz]. C13NMR
(DMSO-d6): 20.8 (CH3), 118.9, 121.3 (CHCO), 127.9, 128.3, 128.7,
129.0, 129.3, 132.0, 133.3, 133.5 (CeCl), 136.7, 137.6, 143.5, 145.1
(HC]CH), 185.4 (C]O). MS m/z (%): 411 (Mþ) (0.44), 92.45 (100).
Anal. Calcd. For C22H18ClNO3S (411): C, 64.15; H, 4.40; N, 3.40.
Found: C, 64.35; H, 4.20; N, 3.70.
4.1.12.5. (E)-N-(4-(3-(4-nitrophenyl)acryloyl)phenyl)-4-
methylbenzenesulfonamide (18). Yield, 46.13%; m.p. 245e247 ꢁC. IR
(KBr, cmꢀ1): 3361 (NH), 3091 (CH arom.), 2933, 2848 (CH aliph.),
1652 (C]O), 1320, 1125 (SO2). H1NMR (DMSO-d6): 2.0 [s, 3H, CH3
tolyl], 7.1, 7.3 [2d, 4H, AreH AB system, J ¼ 6.3 Hz], 7.8 [d, 1H, CHCO,
J ¼ 7.8 Hz], 7.9, 8.0 [2d, 4H, AreH AB system, J ¼ 7.8 Hz], 8.0 [s, 1H,
SO2NH exchangeable with D2O], 8.06 [d, 1H, HC]CH, J ¼ 7.5 Hz],
8.2, 8.3 [2d,4H, AreH AB system, J ¼ 7.8 Hz]. C13NMR (DMSO-d6):
20.8 (CH3), 119.2, 119.5 (CHCO), 123.5, 126.17, 128.3, 129.1, 129.5,
130.19, 136.7, 138.0, 139.0, 141.9, 143.8 (HC]CH), 147.5 (CeNO2),
185.2 (C]O). MS m/z (%): 422 (Mþ) (1.01), 58.60 (100). Anal. Calcd.
For C22H18N2O5S (422): C, 62.55; H, 4.29; N, 6.63. Found: C, 62.45;
H, 4.49; N, 6.83.
4.1.12. General procedure for preparation of compounds (14e19)
To stirring solution of 3b [40] (0.3 g, 0.001038 mol) in ethanol
(15 mL) containing KOH (1.2 g), aromatic benzaldheydes, namely 4-
methyl, 4- fluoro, 4-chloro, 4-nitro, and 4- bromobenzaldehyde
(0.001 mol) were added, the reaction mixture was stirred at room
temperature for 24 h. The precipitated solid was obtained by
filtration and the products were recrystallized from dioxane to give
14e19, respectively.
4.1.12.1. (E)-N-(4-cinnamoylphenyl)-4-methylbenzenesulfonamide
(14). Yield, 97.36%; m.p. >300 ꢁC. IR (KBr, cmꢀ1): 3268 (NH), 3055
(CH arom.), 2921, 2851 (CH aliph.), 1669 (C]O), 1399, 1127 (SO2).
H1NMR (DMSO-d6): 2.08 [s, 3H, CH3tolyl], 7.1, 7.3[2d, 4H, AreH AB
system, J ¼ 6.3 Hz], 7.5 [d, 1H, CHCO, J ¼ 7.9 Hz], 7.5e7.8 [m, 5H,
AreH], 8.0 [s, 1H, SO2NH exchangeable with D2O], 8.06 [d, 1H, HC]
CH, J ¼ 12 Hz] 8.0, 8.3 [2d, 4H, AreH AB system, J ¼ 7.2 Hz]. C13NMR
4.1.12.6. (E)-N-(4-(3-(4-bromophenyl)acryloyl)phenyl)-4-
methylbenzenesulfonamide (19). Yield, 85.52%; m.p. 107e109 ꢁC. IR
(KBr, cmꢀ1): 3360 (NH), 3095 (CH arom.), 2937, 2840 (CH aliph.),
1647 (C]O), 1350, 1134 (SO2). H1NMR (DMSO-d6): 2.0 [s, 3H, CH3

690
M.M. Ghorab et al. / European Journal of Medicinal Chemistry 92 (2015) 682e692
tolyl], 7.1,7.3 [2d, 4H, AreH AB system, J ¼ 6.3 Hz], 7.5 [d, 1H, CHCO,
J ¼ 7.1 Hz], 7.9, 8.0 [2d, 4H, AreH AB system, J ¼ 6.4 Hz], 8.0 [s, 1H,
SO2NH exchangeable with D2O], 8.06 [d, 1H, HC]CH, J ¼ 7.5 Hz],
8.2, 8.3 [2d, 4H, AreH AB system, J ¼ 6.4 Hz]. C13NMR (DMSO-d6):
20.8 (CH3), 119.2, 119.5 (CHCO), 122.9 (CeBr), 126.1, 128.3, 128.5,
129.77, 130.8, 131.6, 134.5, 136.7, 139.0, 141.9, 143.8 (HC]CH), 185.6
(C]O). MS m/z (%): 455 (Mþ) (0.19), 51.10 (100). Anal. Calcd. For
512 (M þ 2) (2.43), 438.70 (100). Anal. Calcd. For C28H22N4O4S
(510): C, 65.87; H, 4.34; N, 10.97. Found: C, 65.64; H, 4.55; N, 11.30.
4.1.16. N-(4-(5-(4-nitrophenyl)-1-carbothioamide-1H-pyrazole-3-
yl)phenyl)-4-methyl-benzenesulfonamide (23)
A mixture of compound 18 (0.3 g, 0.000738 mol), thio-
semicarbazide (0.067 g, 0.000738 mol) and 1.2 g KOH in ethanol
(15 mL) and 5 drops TEA was refluxed for 15 h, then left to cool and
poured onto ice/water and neutralized with dilute hydrochloric
acid. The precipitated was filtered and recrystallized from ethanol
to give 23. Yield, 54.3%; m.p. >300 ꢁC. IR (KBr, cmꢀ1): 3390, 3342,
3286 (NH2, NH), 3184 (CH arom.), 2935, 2830 (CH aliph.), 1310 (C]
S), 1305, 1120 (SO2). H1NMR(DMSO-d6): 2.0 [s, 3H, CH3 tolyl], 2.08
[s, 2H, NH2 exchangeable with D2O], 7.1, 7.3 [2d, 4H, AreH AB sys-
tem, J ¼ 6.0 Hz], 7.2 [s, 1H, CH-pyrazol], 7.6, 8.0 [2d, 4H, AreH AB
system, J ¼ 6.0 Hz], 8.2, 8.3 [2d, 4H, AreH AB system, J ¼ 6.0 Hz ], 8.0
[s, 1H, SO2NH exchangeable with D2O]. C13NMR (DMSO-d6): 20.8,
103.5 (CH-pyrazole), 123.5, 125.9, 127.8, 128.2, 128.3, 129.3, 133.8,
136.2, 137.6, 139.7, 140.0, 144.8 (C]CeN), 147.9, 152.3 (C]N), 176.8
(C]S). MS m/z (%): 493(Mþ) (9.67), 260.05 (100). Anal. Calcd. For
C
22H18BrNO3S (455): C, 57.90; H, 3.98; N, 3.07. Found: C, 57.60; H,
4.10; N, 3.27.
4.1.13. N-(4-((1Z, 2E)-1-(hydroxyimino)-3-(4-nitrophenyl)allyl)
phenyl)-4-methyl-benzene-sulfonamide (20)
A mixture of compound 18 (0.3 g, 0.00073 mol), hydroxylamine
(0.05 g, 0.00073 mol) and 1.2 g KOH in ethanol (15 ml) was refluxed
for 10 h, then cold and poured onto ice/water and neutralized with
dilute hydrochloric acid. The precipitated was filtered and recrys-
tallized from ethanol to give 20. Yield, 63.76%; m.p. > 300 ꢁC. IR
(KBr, cmꢀ1): 3451 (OH), 3265 (NH), 3015 (CH arom.), 2926, 2829
(CH aliph.), 1350, 1115 (SO2). H1NMR (DMSO-d6): 2.0 [s, 3H, CH3
tolyl], 2.3 [s, 1H, OH exchangeable with D2O], 6.6 [d, 1H, CH]CH,
J ¼ 6.4 Hz], 6.63 [d, 1H, CHeC6H4, J ¼ 6.9 Hz], 7.0, 7.1 [2d, 4H, AreH
AB system, J ¼ 7.1 Hz], 7.3, 7.5 [2d, 4H, AreH AB system, J ¼ 6.0 Hz],
8.0, 8.2 [2d, 4H, AreH AB system, J ¼ 6.0 Hz], 8.5 [s, 1H, SO2NH
exchangeable with D2O]. C13NMR (DMSO-d6): 20.8, 122.3, 123.8,
127.9, 128.0, 128.6, 129.5, 129.8, 131.2, 134.0, 135.5, 142.3, 142.5,
142.6, 147.1, 164.8 (C]N). MS m/z (%): 437 (Mþ) (20.23), 58.5 (100).
Anal. Calcd. For C22H19N3O5S (437): C, 60.40; H, 4.38; N, 9.61.
Found: C, 60.21; H, 4.54; N, 9.43.
C23H19N5O4S2 (493): C, 55.97; H, 3.88; N, 14.19. Found: C, 55.74; H,
4.10; N, 14.34.
4.1.17. N-(4-(-5-(4-nitrophenyl)-1-hydrazinecarbonothioyl-1H-
pyrazol-3-yl)phenyl)-4-methylbenzenesulfonamide (24)
A mixture of compound 18 (0.5 g, 0.00123 mol), thiocarbohy-
drazide (0.13 g, 0.00123 mol) and 1.2 g KOH in ethanol (15 mL) and
5 drops TEA was refluxed for 15 h, then left to cool and poured onto
ice/water and neutralized with dilute hydrochloric acid. The
precipitated was filtered and recrystallized from ethanol to give 24.
Yield, 56%; m.p. 163e165 ꢁC. IR (KBr, cmꢀ1): 3398, 3305, 3213 (NH2,
2NH), 3060 (CH arom.), 2928, 2830 (CH aliph.), 1320 (C]S), 1305,
1150 (SO2). H1NMR (DMSO-d6): 2.0 [s, 3H, CH3 tolyl], 3.89 [s, 3H,
NH2, NH exchangeable with D2O], 7.1, 7.3 [2d, 4H, AreH AB system,
J ¼ 6.0 Hz], 7.6, 7.7 [2d, 4H, AreH AB system, J ¼ 6.0 Hz], 7.8 [s, 1H,
CH-pyrazol], 7.8, 7.83 [2d, 4H, AreH AB system, J ¼ 6.0 Hz], 8.0 [s,
1H, SO2NH exchangeable with D2O]. C13NMR (DMSO-d6): 20.8,
104.5 (CH-pyrazol), 119.4, 125.0, 126.4, 127.8, 127.9, 129.6, 131.4,
134.5, 136.2, 139.2, 139.7, 144.3 (C]CeN), 147.6, 152.3 (C]N), 174.9
(C]S). MS m/z (%): 508(Mþ) (19.24), 58.26 (100). Anal. Calcd. For
4.1.14. N-(4-(5-(4-nitrophenyl)-1H-pyrazol-3-yl)phenyl)-4-
methylbenzenesulfonamide (21)
A mixture of compound 18 (0.3 g, 0.000738 mol), hydrazine
hydrate (0.036 g (0.034 ml), 0.000738 mol) and 1.2 g KOH in
ethanol (15 mL) and 5 drops TEA was refluxed for 15 h, then left to
cool and poured onto ice/water and neutralized with dilute hy-
drochloric acid. The precipitated was filtered and recrystallized
from ethanol to give 21. Yield, 62.5%; m.p. > 300 ꢁC. IR (KBr, cmꢀ1):
3343, 3249 (2NH), 3199 (CH arom.), 2918, 2830 (CH aliph.), 1336,
1120 (SO2). H1NMR (DMSO-d6): 2.0 [s, 3H, CH3 tolyl], 7.1, 7.3[2d, 4H,
AreH AB system, J ¼ 6.0 Hz], 7.2 [s, 1H, CH-pyrazol], 7.6, 8.0 [2d, 4H,
AreH AB system, J ¼ 6.0 Hz], 8.0 [s, 1H, SO2NH exchangeable with
D2O], 8.2, 8.3 [2d, 4H, AreH AB system, J ¼ 6.0 Hz], 10.3 [s, 1H, NH
exchangeable with D2O]. C13NMR (DMSO-d6): 20.8, 99.7 (CH pyr-
azol), 124.4, 126.2, 127.8, 128.3, 128.8, 129.3, 134.2, 136.2, 137.6,
139.1, 139.7, 147.1 (C]CeN), 147.7 (C]N pyrazol), 147.9. MS m/z (%):
434 (Mþ) (19.82), 58.4 (100). Anal. Calcd. For C22H18N4O4S (434): C,
60.82; H, 4.18; N, 12.90. Found: C, 61.01; H, 4.32; N, 12.74.
C23H20N6O4S2 (508): C, 54.32; H, 3.96; N, 16.52. Found: C, 54.53; H,
4.20; N, 16.32.
4.1.18. N-(4-(6-(4-nitrophenyl)-2-oxo-1,2-dihydropyrimidin-4-yl)
phenyl)-4-methyl-benzenesulfonamide (25)
4.1.15. N-(4-(5-(4-nitrophenyl)-1-phenyl-1H-pyrazol-3-yl)phenyl)-
4-methylbenzenesulfonamide (22)
A mixture of compound 18 (0.3 g, 0.000738 mol), urea (0.044 g,
0.000738 mol) and 1.2 g KOH in ethanol (15 ml) and 5 drops TEA
was refluxed for 9 h, then left to cool and poured onto ice/water and
neutralized with dilute hydrochloric acid. The precipitated was
filtered and recrystallized from ethanol to give 25. Yield, 54.5%;
m.p. 167e169 ꢁC. IR (KBr, cmꢀ1): 3382, 3217 (2NH), 3015 (CH
arom.), 2919, 2825 (CH aliph.), 1669 (C]O), 1310, 1150 (SO2).
H1NMR (DMSO-d6): 2.0 [s, 3H, CH3 tolyl], 6.0 [s, 1H, CH-
pyrimidine], 6.9, 7.0 [2d, 4H, AreH AB system, J ¼ 6.8 Hz], 8.0 [s,
1H, SO2NH exchangeable with D2O], 8.0, 8.1[2d, 4H, AreH AB sys-
tem, J ¼ 6.3 Hz], 8.2, 8.3[2d, 4H, AreH AB system, J ¼ 7.2 Hz], 8.5 [s,
1H, NH exchangeable with D2O]. C13NMR (DMSO-d6): 20.8, 106.6
(CH-pyrimidine), 113.5, 117.9, 121.7, 126.7, 128.3, 129.3, 133.9, 136.4,
136.7, 137.6, 137.8, 147.1 (CeNO2), 156.3 (C]O), 163.3 (C]CeNH),
164.6 (C]N). MS m/z (%): 461 (Mꢀ1) (0.57), 91.05 (100). Anal. Calcd.
For C23H18N4O5S (462): C, 59.73; H, 3.92; N, 12.11. Found: C, 59.55;
H, 3.74; N, 12.34.
A mixture of compound 18 (0.3 g, 0.000738 mol), phenyl hy-
drazine (0.078 g, ml, 0.000738 mol) and 1.2 g KOH in ethanol
(15 mL) and 5 drops TEA was refluxed for 15 h, then left to cool and
poured onto ice/water and neutralized with dilute hydrochloric
acid. The precipitated was filtered and recrystallized from ethanol
to give 22. Yield, 53.7%; m.p. >300 ꢁC. IR (KBr, cmꢀ1): 3354 (NH),
3198 (CH arom.), 2939, 2829 (CH aliph.), 1340, 1120 (SO2). H1NMR
(DMSO-d6): 2.0 [s, 3H, CH3 tolyl], 7.1, 7.3 [2d, 4H, AreH AB system,
J ¼ 6.0 Hz], 7.2 [s, 1H, CH-pyrazol], 7.5, 7.6 [m, 5H, AreH], 7.6, 8.0
[2d, 4H, AreH AB system, J ¼ 6.0 Hz], 8.0 [s, 1H, SO2NH
exchangeable with D2O], 8.2, 8.3 [2d, 4H, AreH AB system,
J ¼ 6.0 Hz]. C13NMR (DMSO-d6): 20.0, 106.7 (CH-pyrazol), 123.7 (CH
phenyl), 124.4, 126.2, 126.4 (CH phenyl), 126.5, 127.8, 128.7, 129.3
(CH phenyl), 129.5, 131.2, 134.2, 137.5, 139.1, 139.7, 139.9 (CH
phenyl), 144.5 (C]CeN), 147.9, 153.4 (C]N pyrazol). MS m/z (%):

M.M. Ghorab et al. / European Journal of Medicinal Chemistry 92 (2015) 682e692
691
4.1.19. N-(4-(6-(4-nitrophenyl)-2-thioxo-1,2-dihydropyrimidin-4-
yl)phenyl)-4-metyhyl-benzenesulfonamide (26)
cancers as in acute leukemia's Hodgkin's disease, and other lym-
phomas and cancers of the breast, adrenal cortex, endometrium,
lung, ovary, and other sites [52].
A mixture of compound 18 (0.3 g, 0.000738 mol), thiourea
(0.056 g, 0.000738 mol) and 1.2 g KOH in ethanol (15 ml) and 5
drops TEA was refluxed for 15 h, then left to cool and poured onto
ice/water and neutralized with dilute hydrochloric acid. The
precipitated was filtered and recrystallized from ethanol to give 26.
Yield, 31.25%; m.p. 180e182 ꢁC. IR (KBr, cmꢀ1): 3383, 3217 (2NH),
3128 (CH arom.), 2919, 2830 (CH aliph.), 1300 (C]S), 1320, 1115
(SO2). H1NMR (DMSO-d6): 2.0 [s, 3H, CH3 tolyl], 6.7 [s, 1H, CH-
pyrimidine], 6.9, 7.0 [2d, 4H, AreH AB system, J ¼ 7.5 Hz], 8.0 [s,
1H, SO2NH exchangeable with D2O], 8.0, 8.1 [2d, 4H, AreH AB
system, J ¼ 7.5 Hz], 8.2, 8.3 [2d, 4H, AreH AB system, J ¼ 7.3 Hz],13.7
[s, 1H, NH exchangeable with D2O]. C13NMR (DMSO-d6): 20.8, 104.3
(CH-pyrimidine), 117.9, 123.3, 126.5, 126.7, 129.5, 129.8, 131.8, 134.8,
136.3, 136.3, 142.2, 143.6, 165.3 (C]N pyrimidine), 176.4 (C]
CeNH), 180.4 (C]S). MS m/z (%): 479 (M þ 1) (7.34), 60.75 (100).
Anal. Calcd. For C23H18N4O4S2 (478): C, 57.73; H, 3.79; N, 11.71.
Found: C, 57.53; H, 3.94; N, 11.54.
4.3. Radisensitizing evaluation
Irradiation was performed in the National Center for Radiation
Research and Technology, Atomic Energy Authority, using Gamma
cell-40 (137Cs) source. The most active compounds 5a, 6a, 6b, 8, 9,
13, 18 and 19 were selected to be reevaluated for their in-vitro
cytotoxic activity in combination with
g
-irradiation. Cells were
plated in 96-multiwell plate (104cells/well) for 24 h before
g-irra-
diation with a single dose of 8 Gy. Cells were incubated for 48 h at
37 ꢁC in atmosphere of 5% CO2. After 48 h, cells were fixed, washed
and stained with 0.4% (wt/vol) SRB dissolved in 1% acetic acid, for
30 min. Excess unbound dye was removed by four washes with 1%
acetic acid and attached stain was recovered with TriseEDTA buffer.
Color intensity was measured in an ELISA reader (Microplate
reader; Sunostik; SPR-960B; China) at a wave length of 570 nm. In
another multiwell plate, cells were incubated with the previously
mentioned selected compounds; compounds 5a, 6a, 6b, 8, 9, 13, 18,
4.2. In-vitro anticancer evaluation against human tumor liver
cancer (HEPG2)
and 19 in molar concentrations of 5, 12.5, 25, 50
mM. After 2 h, cells
were subjected to a single dose of -radiation at a dose level of 8 Gy
g
The in-vitro anticancer activity was measured for twenty six
new compounds on liver human tumor cell line (HEPG2) using the
Sulfo-Rhodamine-B stain (SRB) assay. The in-vitro anticancer
screening was done by the pharmacology unit at the National
Cancer Institute, Cairo University.
with a dose rate of 0.758 rad/sec for 17.73 min, and then the
cytotoxicity was measured 48 h, after irradiation. The surviving
fractions were measured using the above mentioned procedures by
ELISA reader. The surviving fractions were expressed as mean
values
standard error. The results were analyzed using 1-way
The sulfo-rhodamine B (SRB) assay which was developed in
1990 [50], remains one of the most widely used methods for in-
vitro cytotoxic screening. The assay relies on the ability of SRB to
bind to protein components of cells that have been fixed to tissue-
culture plates by trichloroacetic acid (TCA). SRB is a bright-pink
aminoxanthene dye with two sulfonic groups that bind to basic
amino-acid residues under mild acidic conditions, and dissociate
under basic conditions. The amount of dye extracted from stained
cells is directly proportional to the cell mass [51]. Cells were plated
in 96-multiwell plate (104 cells/well) for 24 h before treatment
with the compounds to allow attachment of cell to the wall of the
plate. Test compounds were dissolved in dimethylsulfoxide (DMSO)
and diluted with saline to the appropriate volume. Different con-
ANOVA test and given in Table 2.
Appendix A. Supplementary data
Supplementary data related to this article can be found at http://
dx.doi.org/10.1016/j.ejmech.2015.01.036.
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