Antibiotics · Antimycotics · Antiparasitics · Antiviral Drugs · Chemotherapeutics · Cytostatics

Synthesis of some new pyrazole and pyrimidine derivatives

carrying a sulfonamide moiety of expected antitumor
activity and study of the synergistic effect of g-irradiation
Mostafa M. Ghorab1, Fatma A. Ragab2, Helmy I. Heiba1, Hanan A. Youssef2, Marwa Galal1
1
Department of Drug Radiation Research, National Center for Radiation Research and Technology, Atomic Energy Authority, Nasr
City, Cairo, Egypt
2
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Cairo University, Cairo, Egypt

Correspondence to: Mostafa M. Ghorab, Professor of Applied Organic Chemistry, Department of Drug Radiation Research, National
Center for Radiation Research and Technology, Atomic Energy Authority, P.O. Box 29, Nasr City, Cairo, Egypt;

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e-mail: mmsghorab@yahoo.com

Key words Abstract

n c-Irradiation Several novel pyrazole (11 – 18) and pyri- equipotent while the others were more
n Pyrazole derivatives midine (19 – 23) derivatives were synthe- potent than doxorubicin (CAS 25316-40-9).
n Pyrimidine derivatives sized starting from different sulfona- The synergistic effect with c-radiation
mides and different active methylenes. was also studied.
n Sulfonamides, antitumor
The synthesized compounds were char-
activity
acterized by elemental analysis, IR,
1
H-NMR and mass spectral data. The ob-
Arzneimittelforschung tained compounds were screened as anti-
2010;60(1):48–55 tumor agents against human tumor cell
line. Some of the tested compounds were

1. Introduction 2400 analyser (Perkin-Elmer, Norwalk, CT, USA) at the micro-

Sulfonamides constitute an important class of drugs
having several pharmacological activities including anti-
bacterial [1, 2], antiviral [3, 4], antifungal [5, 6], hypogly-
cemic [7, 8], diuretic [9, 10] and anticancer activities
[11 – 15]. They exhibit their anticancer activities through
different mechanisms of action, the most important
being carbonic anhydrase inhibition [16, 17]. In addi-
tion, several heterocyclic compounds incorporating
pyrazole [18, 19] or pyrimidine [20, 21] rings showed sig-
nificant antitumor activities. In this work benzene-
sulfonamides were used as starting materials to react
with several active methylenes to yield novel pyrazole
and pyrimidine derivatives having a sulfonamide moiety
hoping that these new compounds show significant an-
ticancer activities.
analytical laboratories of the Faculty of Science, Cairo Univer-
sity. All compounds were within ± 0.4 % of the theoretical val-
ues. The IR spectra (KBr) were measured on a Shimadzu IR
110 spectrophotometer (Shimadzu, Koyoto, Japan), 1H-NMR
spectra were obtained on a Bruker proton NMR-Avance 300
(300 MHz) (Bruker, Munich, Germany) with DMSO-d6 as a sol-
vent, using tetramethylsilane (TMS) as internal standard. Mass
spectra were run on HP Model MS-5988 (Hewlett Packard,
Palo, Alto, California, USA). All reactions were monitored by
thin layer chromatography (TLC) using precoated aluminium
sheets silica gel Merck 60 F254 and were visualized by a UV
lamp (Merck, Darmstadt, Germany). Physico-chemical and
analytical data of the synthesized compounds 4 – 23 are pre-
sented in Table 1.
2.2 Synthesis

2. Materials and methods
2.1 Chemistry
Melting points are uncorrected and were determined on a
Stuart melting point apparatus (Stuart Scientific, Redhill, UK).
Elemental analyses (C, H, N) were performed on Perkin-Elmer
2.2.1 4-(2,2-Dicyanovinylamino)benzenesulfonamide
(4), N-[4-(2,2-dicyanovinylamino)phenylsulfonyl]acet-
amide (6), 4-(2,2-dicyanovinylamino)-N-(3-methyl-
isoxazol-5-yl)benzenesulfonamide (9)
A mixture of sulfonamide (sulfanilamide, sulfacatamide, sulfa-
methoxazole) (0.01 mol), malononitrile (0.01 mol), triethyl-
orthoformate (0.01 mol) and acetic acid (1 ml) in methanol

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48 Ghorab et al. – Pyrazoles and pyrimidines
 Antibiotics · Antimycotics · Antiparasitics · Antiviral Drugs · Chemotherapeutics · Cytostatics

Table 1: Physico-chemical and analytical data of compounds 2.2.3 N-[4-(2-Acetyl-3-oxobut-1-enylamino)phenyl-

4 – 23. sulfonyl]acetamide (8)
Compd. M. p. (oC) Yield (%) Mol. formula A mixture of sulfacetamide (0.01 mol), acetylacetone (0.01 mol),
4 222 – 223 63 C10H8N4O2S
triethylorthoformate (0.01 mol) and acetic acid (1 ml) in
5 185 – 187 60 C12H13N3O4S methanol (30 ml) was refluxed for 5 h. The reaction mixture
6 252 – 254 58 C12H10N4O3S was filtered and recrystallized from dioxane.
7 178 – 179 70 C14H15N3O5S
IR (KBr, cm–1) (8): 3486, 3400 (NH), 3042 (CH arom.), 2965,
8 200 – 201 80 C14H16N2O5S
9 229 – 230 60 C14H11N5O3S 2834 (CH aliph.), 1720 (C=O), 1306, 1156 (SO2).
10 210 – 211 66 C16H16N4O5S d: 1.9, 2.0, 2.1 [3s, 9H, 3COCH3], 5.6 [d, 1H, CH], 5.8 [s, 1H,
11 140 – 142 54 C12H13N5O4S NH], 7.3, 7.7 [2d, 4H, Ar-H AB system], 8.4 [s, 1H, SO2NH].
12 > 280 66 C14H14N6O4S
13 61 – 62 55 C16H16N6O2S
14 70 – 72 60 C18H18N6O3S
15 199 – 200 90 C18H17N5O4S
2.2.4 N-{4-[(3-Amino-5-oxo-1H-pyrazol-4(5H)-ylidene)
16 71 – 72 50 C20H19N7O3S methylamino]phenylsulfonyl}acetamide (11), 4-[(3-
17 220 – 221 60 C20H18N6O4S Amino-5-oxo-1,5-dihydropyrazol-4-ylidenemethyl)
18 160 – 162 49 C20H22N4O3S
19 120 – 122 68 C17H12N4O3S2 amino]benzenesulfonamide (12)
20 200 – 202 70 C19H14N4O4S2 A mixture of 7 or 10 (0.01 mol) and hydrazine hydrate
21 212 – 214 60 C21H15N5O4S2
22 > 280 95 C11H11N5O3S2 (0.01 mol) in dioxane (20 ml) was refluxed for 5 h. The reaction

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23 222 – 224 62 C15H16N4O3S2
(30 ml) was refluxed for 5 h. The reaction mixture was filtered
and recrystallized from ethanol.
IR (KBr, cm–1) (4): 3348, 3272, 3218 (NH, NH2), 3074 (CH
arom.), 2960, 2890 (CH aliph.), 2220 (CN), 1310, 1146, (SO2).
MS (m/z): 248 (100 %), 231 (34.12 %), 168 (20.70 %), 141
(41.12 %).
IR (KBr, cm–1) (6): 3376, 3190 (NH), 3076 (CH arom.), 2966,
2845 (CH aliph.), 2224 (CN), 1706 (C=O), 1330, 1160 (SO2).
d: 2.4 [s, 3H, COCH3], 6.2 [d, 1H, CH], 6.6 [s, 1H, NH], 7.8, 8.0
[2d, 4H, Ar-H AB system], 8.5 [s, 1H, SO2NH].
IR (KBr, cm–1) (9): 3368, 3294 (NH), 3082 (CH arom.), 2980,
2890 (CH aliph.), 2222 (CN), 1320, 1160 (SO2).
d: 2.3 [s, 3H, CH3 isoxazole], 6.1 [d, 1H, CH], 6.5 [s, 1H, NH],
7.5, 7.8 [2d, 4H, Ar-H AB system], 8.5 [s, 2H, CH isoxazole +
SO2NH].
2.2.2 Ethyl 2-cyano-3-(4-sulfamoylphenylamino)
acrylate (5), ethyl 3-[4-(N-acetylsulfamoyl)phenyl-
amino]-2-cyanoacrylate (7), ethyl 2-cyano-3-
{4-[N-(3-methylisoxazol-5-yl)sulfamoyl]phenylamino}
acrylate (10)
A mixture of sulfonamide (sulfanilamide, sulfacetamide, sulfa-
methoxazole) (0.01 mol), ethylcyanoacetate (0.01 mol), triethyl-
orthoformate (0.01 mol) and acetic acid (1 ml) in methanol
(30 ml) was refluxed for 5 h. The reaction mixture was filtered
and recrystallized from ethanol.
IR (KBr, cm–1) (5): 3350, 3270 (NH, NH2), 3090 (CH arom.),
2976, 2887 (CH aliph.), 2216 (CN), 1700 (C=O), 1308, 1152 (SO2).
d: 1.3 [t, 3H, CH3], 4.2 [q, 2H, CH2], 6.5 [d, 1H, CH], 6.8 [s, 1H,
NH], 7.4, 7.8 [2d, 4H, Ar-H AB system], 8.4 [s, 2H, SO2NH2].
IR (KBr, cm–1) (7): 3390, 3248 (NH), 3020 (CH arom.), 2985,
2877 (CH aliph.), 2220 (CN), 1652 (C=O), 1312, 1156 (SO2).
d: 1.2 [t, 3H, CH3], 2.2 [s, 3H, COCH3], 4.2 [q, 2H, CH2], 6.1 [d,
1H, CH], 6.5 [s, 1H, NH], 7.4, 7.8 [2d, 4H, Ar-H AB system], 8.4
[s, 1H, SO2NH].
IR (KBr, cm–1) (10): 3350, 3152 (NH), 3090 (CH arom.), 2960,
2889 (CH aliph.), 2218 (CN), 1716 (C=O), 1320, 1164 (SO2).
d: 1.2 [t, 3H, CH3], 2.3 [s, 3H, CH3 isoxazole], 4.2 [q, 2H, CH2],
6.1 [d, 1H, CH], 6.8 [s, 1H, NH], 7.5, 7.8 [2d, 4H, Ar-H AB sys-
tem], 8.2 [s, 1H, CH isoxazole], 8.4 [s, 1H, SO2NH].
mixture was cooled and poured into ice water. The solid pro-
duct was filtered and recrystallized from methanol to give 11
and 12, respectively.
IR (KBr cm–1) (11): 3476, 3384, 3258 (NH, NH2), 3086 (CH
arom.), 2990, 2887 (CH aliph.), 1686 (3C=O), 1330, 1158 (SO2).
MS (m/z): 323 (M+, 9 %), 253 (30 %), 156 (88 %), 108 (95 %), 92
(100 %).
IR (KBr cm–1) (12): 3438 (br, NH, NH2), 3100 (CH arom.),
2965, 2835 (CH aliph.), 1684 (2C=O), 1320, 1150 (SO2).
MS (m/z): 362 (M+, 5 %), 256 (70 %), 212 (97 %), 156 (85 %), 90
(100 %).
2.2.5 4-[(3-Amino-5-imino-1-phenyl-1H-pyrazol-
4(5H)-ylidene)methylamino]-benzenesulfonamide
(13), N-{4-[(3-amino-5-imino-1-phenyl-1H-pyrazol-
4(5H)-ylidene)methylamino]phenylsulfonyl}acetamide
(14), 4-[(3-amino-5-imino-1-phenyl-1H-pyrazol-
4(5H)-ylidene)methylamino]-N-(3-methylisoxazol-5-
yl)benzenesulfonamide (15), N-{4-[(3-amino-5-oxo-1-
phenyl-1H-pyrazol-4(5H)-ylidene)methylamino]-phe-
nylsulfonyl}acetamide (16), 4-[(3-amino-5-oxo-1-
phenyl-1,5-dihydro-pyrazol-4-ylidenemethyl)amino]-
N-(3-methyl-isoxazol-5-yl)benzenesulfonamide (17)
A mixture of 4, 6, 7, 9 and/or 10 (0.01 mol) and phenylhydra-
zine (0.01 mol) in dioxane (20 ml) was refluxed for 5 h. The re-
action mixture was cooled and poured into ice water. The solid
product was filtered and recrystallized from ethanol to give 13 –
17, respectively.
IR (KBr cm–1) (13): 3340, 3294, 3222 (NH, NH2), 2934, 2833
(CH aliph.), 3036 (CH arom.), 1336, 1123 (SO2).
MS (m/z): 356 (M+, 22 %), 356 (38 %), 243 (24 %), 201 (22 %),
76 (100 %).
IR (KBr cm–1) (14): 3472, 3380, 3248 (NH, NH2), 3108 (CH
arom.), 2967, 2845 (CH aliph.), 1690 (C=O), 1322, 1150 (SO2).
MS (m/z): 398 (M+, 3.9 %), 253 (22 %), 156 (85 %), 108 (80 %),
92 (100 %).
IR (KBr cm–1) (15): 3424, 3422, 3224 (NH, NH2), 3030 (CH
arom.), 2989, 2823 (CH aliph.), 1320, 1160 (SO2).
MS (m/z): 437 (M+, 17.81 %), 254 (19.18 %), 219 (21.92 %), 161
(26.03 %), 76 (100 %).
IR (KBr cm–1) (16): 3298, 3270, 3260 (NH, NH2), 2980, 2854
(CH aliph.), 3080 (CH arom.), 1734 (3C=O), 1322, 1150 (SO2).
MS (m/z): 399 (M+, 16 %), 378 (50 %), 278 (96 %), 155 (85 %),
127 (100 %).

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Ghorab et al. – Pyrazoles and pyrimidines 49
Antibiotics · Antimycotics · Antiparasitics · Antiviral Drugs · Chemotherapeutics · Cytostatics

IR (KBr cm–1) (17): 3420, 3352, 3256 (NH, NH2), 2989, 2845 IR (KBr cm–1) (23): 3460, 3245 (NH), 3080 (CH arom.), 2967,
(CH aliph.), 3032 (CH arom.), 1690 (2C=O), 1316, 1150 (SO2). 2854 (CH aliph.), 1720 (C=O), 1330, 1156 (SO2), 1230 (C=S).
MS (m/z): 438 (M+, 3.9 %), 300 (60 %), 235 (77 %), 145 (86 %), MS (m/z): 364 (M+, 1.15 %), 324 (24.71 %), 267 (21.11 %), 225
90 (100 %). (19.16 %), 201 (100 %).

2.2.6 N-Acetyl-4-[(3,5-dimethyl-1-phenyl-1,5-dihy-
dro-pyrazol-4-ylidenemethyl)amino]benzenesulfon-
amide (18)
A mixture of 8 (0.01 mol) and phenylhydrazine (0.01 mol) in di-
oxane (20 ml) was refluxed for 5 h. The reaction mixture was fil-
tered and recrystallized from ethanol.
IR (KBr cm–1) (18): 3466, 3264 (NH), 3080 (CH arom.), 2954,
2854 (CH aliph.), 1716 (C=O), 1333, 1158 (SO2).
MS (m/z): 398 (M+, 25.49 %), 364(29.41 %), 290 (98 %),
140(41.18 %), 77(100 %).
2.2.7 4-(5-Cyano-4-oxo-3-phenyl-2-thioxo-3,4-di-
hydropyrimidin-1(2H)-yl)benzenesulfonamide (19),
3. Biological testing
3.1 Facilities
The human tumor cell lines (HEPG2 and MCF7) were
available at the National Cancer Institute, Cairo, Egypt.
Irradiation was performed in the National Cancer Insti-
tute, Cairo, Egypt using Gamma cell-40 (60CO) source.
3.2 Measurement of antitumor activity
The antitumor activity of the newly synthesized com-
pounds was measured using the Sulfo-Rhodamine-B
stain (SRB) assay by the method of Skehan and Storeng

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N-[4-(5-cyano-4-oxo-3-phenyl-2-thioxo-3,4-dihydro- [22].
pyrimidin-1(2H)-yl)phenylsulfonyl]acetamide (20), – Cells were plated in 96-multiwell plate (104 cells/well)
4-(5-cyano-4-oxo-3-phenyl-2-thioxo-3,4-dihydropyri- for 24 h before treatment with the compounds to al-
midin-1(2H)-yl)(3-methylisoxazol-5-yl)benzenesulfon- low attachment of cell to the wall of the plate.
amide (21) – Test compounds were dissolved in DMSO and diluted
with saline to the appropriate volume.
A mixture of 4 or 6 and/or 9 (0.01 mol), phenyl isothiocyanate
(0.01 mol) and sodium hydroxide (0.01 mol) in ethanol (20 ml) – Different concentrations of the compounds under test
was refluxed for 3 h. The reaction mixture was cooled, poured (5, 12.5, 25 and 40 lM) were added to the cell mono-
into ice water, and acidified with dil. HCl. The solid product layer. Triplicate wells were prepared for each indivi-
was filtered and recrystallized from dioxane to give 19 – 21, re- dual dose.
spectively. – Monolayer cells were incubated with the compounds
IR (KBr cm–1) (19): 3270, 3200 (NH2), 3016 (CH arom.), 2210 for 48 h at 37 'C and in an atmosphere of 5 % CO2.
(CN), 1686 (C=O), 1310, 1160 (SO2), 1292 (C=S). – After 48 h, cells were fixed, washed and stained for
MS (m/z): 384 (M+, 17.81 %), 374 (26.6 %), 355 (38.1 %), 214 30 min with 0.4 % (wt/vol) with sulforhodamine B
(47.6 %), 92 (100 %).
(SRB) dissolved in 1 % acetic acid.
IR (KBr cm–1) (20): 3382 (NH), 3102 (CH arom.), 2222 (CN),
Unbounded dye was removed by four washes with
1708 (2C=O), 1322, 1156 (SO2), 1234 (C=S).
1 % acetic acid, and the attached stain was recovered
MS (m/z): 426 (M+, 7.78 %), 421 (100 %), 304 (41.47 %), 157
(19.22 %). with Tris-EDTA buffer.
IR (KBr cm–1) (21): 3290 (NH), 3086 (CH arom.), 2222 (CN), – Color intensity was measured in an ELISA reader.
1640 (C=O), 1310, 1164 (SO2), 1230 (C=S). – The relation between surviving fraction and drug con-
MS (m/z): 465 (M+, 20 %), 377 (80 %), 219 (100 %), 202 (70 %), centration was plotted to obtain the survival curve of
105 (90 %), 55 (86 %). each tumor cell line after the specified time.
– The concentration required for 50 % inhibition of cell
2.2.8 4-[(4-Amino-6-oxo-2-thioxo-1,6-dihydro-2H- viability (IC50) was calculated and compared with the
pyrimidin-5-ylidenemethyl)amino]benzenesulfon- reference drug doxorubicin (CAS 25316-40-9). The re-
amide (22) sults are given in Table 2.
A mixture of 5 (0.01 mol), thiourea (0.01 mol) and sodium eth-
oxide (0.01 mol) was refluxed for 5 h. The reaction mixture was Table 2: In vitro cytotoxic activity of some newly synthesized
cooled, poured into ice water, and acidified with dil. HCl The compounds against human liver cell line (HEPG2) and hu-
solid product was filtered and recrystallized from methanol. man breast cell line (MCF7).
IR (KBr cm–1) (22): 3476, 3388, 3300 (NH, NH2), 3080 (CH Compound Cytotoxicity (IC50)a,b (mM)
arom.), 2978, 2836 (CH aliph.), 1680 (C=O), 1322, 1150 (SO2),
1238 (C=S). HEPG2 MCF7
MS (m/z): 325 (M+, 20 %), 200 (59 %), 156 (100 %), 108 (60 %),
11 4.83 5.1
92 (80 %). 13 4.56 3.75
18 4.83 2.95
19 3.22 3.22
2.2.9 N-{4-[(4,6-Dimethyl-2-thioxopyrimidin-5(2H)- 22 3.22 4.02
ylidene)methylamino]phenylsulfonyl}acetamide (23) 23 3.75 5.36
Doxorubicin 5.23 3.22
A mixture of 8 (0.01 mol), thiourea (0.01 mol) and sodium eth-
a
oxide (0.01 mol) was refluxed for 5 h. The reaction mixture was IC50, compound concentration required to inhibit tumor cell
proliferation by 50 %.
cooled, poured into ice water, and acidified with dil. HCl. The b
Values are means of three experiments.
solid product was filtered and recrystallized from ethanol.

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3.3 Measurement of synergism with radiation The cytotoxicity of the control group, radiation
This study was conducted to compare the potency of group, drug treated groups (5, 12.5, 25, 40 mM) and the
the tested compounds as antitumor agents alone or in irradiated drugs groups (5, 12.5, 25, 40 mM) was mea-
combination with radiation to predict the synergistic ef- sured using the SRB assay discussed above on human
fect of g-irradiation, since the studies proved the efficacy liver cell line (HEPG2) and human breast cell line
of combining chemotherapy with radiotherapy for pa- (MCF7).
tients with cancer to decrease the side effects of both
drugs and radiation [23]. 3.4 Statistical analysis
The most potent compounds from the in vitro study
The surviving fraction was expressed as means – SE. The
(11, 13, 18, 19, 22 and 23) were selected to predict the
differences between drug treated groups and the control
effect of combination with radiation against human li-
group was analysed with 2-way ANOVA test and the re-
ver and breast cell lines (HEPG2 and MCF7).
sults are given in Tables 3 and 4. The effect of radiation
A single dose of g-irradiation was delivered at a dose
on cytotoxic activity of different concentrations of com-
level of 4 Gy and a dose rate of 2 Gy/min. Irradiation
pound 19 on HEPG2 cell line is presented in Fig. 1.
was performed at the National Cancer Institute, Cairo,
Egypt using Gamma cell-40 (60CO) source.

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Fig. 1: Effect of radiation on the cytotoxic activity of different concentrations of compound 19 on HEPG2 cell line.

Table 3: In vitro cytotoxic activity of the most potent synthesized compounds against human liver cell line (HEPG2) and human
breast cell line (MCF7).
Drug concentration (mM)

Cpd. Control Control 5 12.5 25 40

Cell line
No (cells only) (radiation)
#
Means – SE (% change)

1.4152 – 0.02606 1.24 – 0.08145ns 0.7367 – .03495* 0.5283 – .03326* 0.2747 – .01768* 0.274 – .02409*
13
(100 %) (12.38 %) (47.9 %) (62.67 %) (80.63 %) (80.6 %)
1.4152 – 0.02606 1.24 – 0.08145ns 0.8747 – 0.06466* 0.622 – 0.01504* 0.266 – 0.02203* 0.2453 – 0.009280*
19 (100 %) (12.38 %) (38.19 %) (56.1 %) (81.2 %) (82.6 %)
HEPG2
1.5139 – 0.03894 1.2303 – 0.04683ns 0.829 – 0.03215* 0.625 – 0.02951* 0.3593 – 0.04999* 0.298 – 0.03081*
22
(100 %) (18.73 %) (45.23 %) (58.7 %) (76.28 %) (80.3 %)

1.526 – 0.05680 1.2022 – 0.03297ns 1.029 – 0.1175* 0.845 – 0.07450* 0.6393 – 0.1084* 0.414 – 0.08489*
23
(100 %) (21.2 %) (32.56 %) (44.62 %) (58.1 %) (72.8 %)

1.3613 – 0.05082 1.261 – 0.03745ns 1.189 – 0.03185ns 0.7203 – 0.04418* 0.3563 – 0.03908* 0.336 – 0.02558*
19
(100 %) (7.3 %) (12.6 %) (47.1 %) (73.8 %) (75.3 %)
MCF7
1.3613 – 0.05082 1.261 – 0.03745ns 0.95 – 0.06048* 0.4113 – 0.03266* 0.3677 – 0.07027* 0.4033 – 0.01627*
18
(100 %) (7.3 %) (30.2 %) (69.8 %) (73 %) (70.3 %)

Each value is the mean of three values – SE.

#
: Percentage of change from control group.
ns
: Non-significant difference from control group at p < 0.05.
*: Significant difference from control group at p < 0.05.

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Table 4: In vitro cytotoxic activity of the most potent synthesized compounds after irradiation against human liver cell line
(HEPG2) and human breast cell line (MCF7).
Drug Irradiated (mM)

Cpd. Control Control

Cell line 5 12.5 25 40
No (cells only) (radiation)
#
Means – SE (% change)

1.4152 – 0.02606 1.24 – 0.08145ns 0.3967 – .08669* 0.1743 – .01067* 0.1267 – .01795* 0.1083 – .01832*
13
(100 %) (12.38 %) (72 %) (87.7 %) (91 %) (92.3 %)

1.4152 – 0.02606 1.24 – 0.08145ns 0.603 – 0.05787* 0.4247 – 0.01915* 0.134 – 0.02768* 0.073 – 0.002000*
19
(100 %) (12.38 %) (57.3 %) (70.3 %) (90.5 %) (94.84 %)
HEPG2
1.5139 – 0.03894 1.2303 – 0.04683ns 0.5297 – 0.08162* 0.2663 – 0.08453* 0.224 – 0.05352* 0.103 – 0.02558*
22
(100 %) (18.73 %) (65 %) (82.4 %) (85.2 %) (93 %)

1.526 – 0.05680 1.2022 – 0.03297ns 0.5387 – 0.01622* 0.44 – 0.07485* 0.1807 – 0.008819* 0.1427 – 0.01746*
23
(100 %) (21.2 %) (64..69 %) (71.2 %) (88 %) (90.6 %)

1.3613 – 0.05082 1.261 – 0.03745ns 0.841 – 0.02030* 0.5277 – 0.05152* 0.2203 – 0.02513* 0.156 – 0.03005*
19
(100 %) (7.3 %) (38.2 %) (61.2 %) (83.8 %) (88.5 %)
MCF7

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1.3613 – 0.05082 1.261 – 0.03745ns 0.7337 – 0.03767* 0.2717 – 0.02598* 0.1163 – 0.02572* 0.2117 – 0.02908*
18
(100 %) (7.3 %) (46 %) (80 %) (91.4 %) (84.5 %)

Each value is the mean of three values – SE.

#
: Percentage of change from control group.
ns
: Non-significant difference from control group at p < 0.05.
*: Significant difference from control group at p < 0.05.

4. Results and discussion (Scheme 4). Its mass spectrum showed a molecular ion
peak at 398. Sulfonamide derivatives were used in the
4.1 Chemistry
synthesis of several novel pyrimidine derivatives bearing
Interaction of different sulfonamides (sulfanilamide, sulfonamides in order to study their structure-activity
sulfacetamide, sulfamethoxazole) with different active relationships and their anticancer activities. Thus, inter-
methylenes (malononitrile, ethylcyanoacetate, acetyl action of compounds 4, 6, 9 with phenyl isothiocyanate
acetone) yielded sulfonamide derivatives 4 – 10, respec- in NaOH/ethanol gave the corresponding pyrimidine
tively (Scheme 1). IR spectra of compounds 4, 6, 9, derivatives 19 – 21 (Scheme 5). Their IR spectra showed
showed the presence of characteristic CN bands, mass the presence of CN and C=O bands, while their mass
spectrum of compound 4 showed a molecular ion peak
spectra showed molecular ion peaks at 384, 426 and
at 248 which is the base peak, IR spectra of compounds
465, respectively. Interaction of compound 5 with
5, 7, 10 exhibited the presence of CN and ester bands,
thiourea in sodium ethoxide yielded the pyrimidine de-
while their NMR spectra showed the presence of triplet
rivative 22 (Scheme 5). The IR spectrum showed disap-
and quartet signals characteristic of the ester group. The
IR spectrum of compound 8 showed the presence of a
C=O band characteristic ofthe acetyl group. In the pre-
sent work, the reactivity of sulfonamide derivatives 4 – 10
against different nucleophiles was studied in order to
obtain biologically active pyrazole and pyrimidine deri-
vatives bearing sulfonamide moieties. Thus, their inter-
action with hydrazine hydrate or phenyl hydrazine in
dioxane as a solvent yielded the corresponding pyrazole
derivatives 11 – 17 (Scheme 2, 3). IR spectra of com-
pounds 8 – 10 showed the disappearance of CN bands
and presence of forked bands characteristic of NH2,
while their mass spectra revealed molecular ion peaks
at 356, 398 and 438, respectively. IR spectra of com-
pounds 11, 12, 15, 17 showed the disappearance of CN
bands and presence of C=O and the forked bands char-
acteristic of NH2, while their mass spectra revealed mo-
lecular ion peaks at 323, 362, 437 and 438, respectively.
Reaction of sulfonamide derivative 8 with phenyl hydra-
zine in dioxane yielded the pyrazole derivative 18 Scheme 1

Arzneimittelforschung 2010;60(1):48–55
52 Ghorab et al. – Pyrazoles and pyrimidines
 Antibiotics · Antimycotics · Antiparasitics · Antiviral Drugs · Chemotherapeutics · Cytostatics

Scheme 6

pearance of CN bands and presence of C=O and forked

bands characteristic of NH2, while the mass spectrum
revealed a molecular ion peak at 325. Finally, reaction
Scheme 2
of sulfonamide derivative 8 with thiourea yielded the
pyrimidine derivative 23 (Scheme 6). Its mass spectrum
showed a molecular ion peak at 364.

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4.2 Antitumor activity
Some of the synthesized compounds were tested for
their antitumor activities in vitro against human liver
and breast cancer cell lines (HEPG2 and MCF7) and
some of the tested compounds were equipotent while
the others were more potent compared with doxorubi-
cin.
For the liver cell line (HEPG2), the pyrimidine deri-
vatives 19, 22 and 23 showed the highest antitumor ac-
tivity with IC50 of 3.22, 3.22 and 3,75 lM, respectively.
On the other hand, the pyrazole derivatives 11, 13 and
18 showed lower activities with IC50 of 4.83, 4.56 and
Scheme 3
4.83 lM, respectively. All the tested compounds showed
higher activity compared with the reference drug doxoru-
bicin with IC50 corresponding to 5.23 lM.
For the breast cell line (MCF7), the pyrazole deriva-
tive 18 exhibited the highest activity with IC50 of
2.95 lM which is found to be more active than doxoru-
bicin (3.22 lM), while the pyrimidine derivative 19
showed cytotoxic activity similar to that of doxorubicin.
On the other hand, the pyrazole derivative 13 showed
Scheme 4 antitumor activity with IC50 of 3.75 lM, while the pyra-

Scheme 5

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Ghorab et al. – Pyrazoles and pyrimidines 53
Antibiotics · Antimycotics · Antiparasitics · Antiviral Drugs · Chemotherapeutics · Cytostatics
zole derivatives 11, 22 and 23 showed lower activities
with IC50 of 5.1, 4.02 and 5.36 lM.
These preliminary results of biological screening of
the tested compounds indicate that the pyrimidine deri-
vatives were more active on the liver cell line (HEPG2),
while the pyrazole derivatives were more active on the
breast cell line (MCF7).
4.3 Synergism with radiation
The results shown in Tables 3 and 4 demonstrate that
the radiation group showed non-significant difference
from the control group. Comparing different compound
concentrations with the control group on HEPG2 the
following was found: For 5, 12.5, 25, 40 (mM) all the
tested compounds showed significant differences from
control before radiation and also showed significant
changes after radiation but with higher percentage of
change from control.
For MCF7 the following was found: At 5 lM com-
pound 19 showed non-significant difference from con-
trol before radiation, while compound 18 showed signif-
icant difference from control. On the other hand, when
irradiated, both compounds showed significant differ-
ences from control.
At 12.5, 25, 40 lM the tested compounds 18 and 19
showed significant differences from control before ra-
diation, while after irradiation the change from control
increased significantly.
5. Conclusion
From the above results, we can conclude that adminis-
tration of the tested compounds on human liver and
breast cell lines (HEPG2 and MCF7) showed promising
anticancer activity, while combining these compounds
with radiation at the same concentrations enhanced
their activity which demonstrates the importance of the
combination therapy for patients with cancer to de-
crease the side effects of both drugs and radiation.
Conflict of interest
The authors declare that, except funding provided by National
Center for Radiation Research and Technology, Atomic Energy
Authority, Cairo, Egypt, no financial support has been received
from any individual for research and there are no personal fi-
nancial holdings that could be perceived as constituting a po-
tential conflict of interest.
54 Ghorab et al. – Pyrazoles and pyrimidines
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