Novel quinazoline derivatives bearing a sulfapyridine moiety as anticancer and radiosensitizing agents

Article abstract of DOI:10.1002/jhet.1830

Quinazoline derivatives posses many types of biological activities and have recently been reported to show substantial antitumor activity in vitro and/or in vivo. There is a variety of mechanisms for their anticancer activity. The present work reports the possible utility of methyl anthranilate in the synthesis of some new quinazoline derivatives, bearing a substituted sulfonamide moiety. All the newly synthesized compounds were evaluated for their in vitro anticancer activity against human liver cancer cell line, using doxorubicin as a reference drug. In addition, the most active compounds 14 and 15 were selected and evaluated for their ability to enhance the cell killing effect of γ-radiation.

Full text of DOI:10.1002/jhet.1830

Month 2014
Novel Quinazoline Derivatives Bearing a Sulfapyridine Moiety as
Anticancer and Radiosensitizing Agents
Mostafa M. Ghorab, * Fatma A. Ragab, Helmi I. Heiba, and Ahmad A. Bayomi
a
b
c
c
a
c
Medicinal, Aromatic and Poisonous Plants Research Center (MAPPRC), College of Pharmacy, King Saud University,
P.O. Box 2457, Riyadh 11451, Saudi Arabia
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Cairo University, Cairo, Egypt
Department of Drug Radiation Research, National Center for Radiation Research & Technology, Atomic Energy Authority,
b
Cairo, Egypt
*
E-mail: mmsghorab@yahoo.com
Received April 21, 2012
DOI 10.1002/jhet.1830
Published online 00 Month 2014 in Wiley Online Library (wileyonlinelibrary.com).
Quinazoline derivatives posses many types of biological activities and have recently been reported to
show substantial antitumor activity in vitro and/or in vivo. There is a variety of mechanisms for their antican-
cer activity. The present work reports the possible utility of methyl anthranilate in the synthesis of some new
quinazoline derivatives, bearing a substituted sulfonamide moiety. All the newly synthesized compounds
were evaluated for their in vitro anticancer activity against human liver cancer cell line, using doxorubicin
as a reference drug. In addition, the most active compounds 14 and 15 were selected and evaluated for their
ability to enhance the cell killing effect of g-radiation.
J. Heterocyclic Chem., 00, 00 (2014).
INTRODUCTION
kinase inhibition [25], tymidylate synthase inhibition [26],
selective erB2 receptor tyrosine kinase inhibition [27],
selective c-Src kinase inhibition [28], and tumor suppressor
protein (p-53) reactivator [29].
According to the data, quinazoline derivatives show
inhibition of the growth of human hepatocelluar cancer
cells [30,31] and have synergistic effect when combined
with g-radiation through potentiatiation of the antitumor
effect of single and multiple fractions of g-radiation [32,33].
Sulfonamides constitute an important class of drugs,
with several types of pharmacological activities including
antibacterial [34], carbonic anhydrase inhibitors [35],
diuretic [36], hypoglycemic [37], and antithyroid activity
Quinazolines constitute an important class of heterocyclic
compounds, which represent building block for approxi-
mately 150 naturally occurring alkaloids [1,2] and have
relieved an increasing interest from medicinal chemists
because of their wide range of biological and pharmaceutical
activities including antidiabetic [3] antihyperglycemic [4],
antihypertensive [5], antihistaminic [6], antioxidant [7]
analgesic, anti-inflammatory [8], antipsychotic, muscle
relaxant [9], anticonvulsant [10],antiparkinsonian [11],
antitoxoplasmic [12], antitubercular [13], antibacterial [14],
antifungal [15], and antiviral [16] activities. Quinazoline
derivatives have been reported to show substantial antitumor
activity in vitro and/or in vivo [17–21].
[38]. Also, some structurally novel sulfonamide derivatives
have recently been reported to show substantial antitumor
activity in vitro and/or in vivo [39–42].
In the light of these facts, we planned to synthesize
novel quinazoline derivatives, in order to study their
structure activity relationship, and hoped that the new
compounds might show significant anticancer activity.
From the literature survey, it was found that quinazoline
derivatives act as antitumor agents through a variety of
mechanisms such as dihydrofolate reductase inhibition [22],
microtubule polymerization inhibition [23], epidermal
growth factor receptor inhibition [24], cyclin-dependent
©
2014 HeteroCorporation

M. M. Ghorab, F. A. Ragab, H. I. Heiba, and A. A. Bayomi
Vol 000
Moreover, we also aimed to evaluate most active com-
pounds for their in vitro anticancer activity in combination
with g-radiation, to evaluate their ability to enhance the
cytotoxic activity of g-radiation.
derivative 5b (lead acetate paper). The IR spectrum
of compound 5 showed bands at 3410, 3308, and
À1
À1
3183 cm (NH, NH ), 1660 cm (C═O), and 1383 and
2
À1
1
1133 cm (SO ). The H NMR spectrum of compound 5
2
in (DMSO-d ) revealed signals at 6.8 ppm assigned to
6
(
NH ) group and 13.1 ppm corresponding to (SO NH)
2
2
RESULTS AND DISCUSSION
group. The mass spectrum of compound 5b revealed a
molecular ion peak m/z at 408 [M ] (2.1%), with a base
peak m/z at 343 (100%).
The reactivity of N-amino derivative 5b was studied.
Thus, interaction of compound 5b with formic acid yielded
the 1,2,4-triazolo[5,1-b]quinazoline derivative 6 (Scheme 2).
The structure of compound 6 was established by elemental
analysis, IR, H NMR, and mass spectroscopy. The IR spec-
trum showed bands at 3227 cm (NH), 1663 cm (C═O),
+
The starting material methyl-2-isothiocyanatobenzoic
acid 2 was synthesized from the reaction of methyl anthra-
nilate with thiophosgene [43]. The reactivity of isothio-
cyanato derivative 2 toward nitrogen nucleophile was
investigated. When compound 2 was reacted with sulfa-
pyridine 3 in dimethylformamide at room temperature, the
thioureido derivative 4 was obtained in good yield
1
À1
À1
(
Scheme 1). The structure of compound 4 was supported
1
À1
1
by elemental analysis, IR, H NMR, and mass spectral data.
and 1395 and 1139cm (SO ). The H NMR spectrum of
2
IR spectrum of compound 4 showed the absence of NCS
compound 6 in (DMSO-d ) revealed signals at 8.0 ppm
6
À1
band and the presence of bands at 3300 and 3238 cm
assigned to CH of triazole ring and a multiplet at
6.9–7.8 ppm for aromatic protons. The mass spectrum of
compound 6 revealed a molecular ion peak m/z at 418
À1
À1
(
1
3NH), 1651 cm (C═O), 1262 cm (C═S), 1389 and
À1
À1
1
133 cm (SO ), and 1262 cm (C═S). The H NMR
2
+
spectrum of compound 4 in (DMSO-d ) revealed signal at
[M ] (1.65%), with a base peak m/z at 88 (100%). When
6
3
.6 ppm because of (OCH ) group. The mass spectrum
compound 5b was refluxed with acetic anhydride for short
time (10 min), the corresponding mono acetyl derivative 7
was obtained whereas for long time (10 h), gave the cyclic
compound triazoloquinazoline 8 (Scheme 2). The struc-
ture of compounds 7 and 8 was established by elemental
3
of compound 4 revealed a molecular ion peak m/z at
4
+
29 [M À CH ] (0.4%), with a base peak m/z at 226 (100%).
3
Treatment of compound 4 with hydrazine hydrate in
ethanol afforded the corresponding N-amino derivative
b. The formation of compound 5b was assumed to occur
1
5
analysis, IR, and H NMR spectral data. The IR spectrum
À1
via elimination of 1 mol H S of intermediate 5a followed
by intramolecular cyclization [44] to give the N-amino
of compound 7 showed bands at 3190 cm (NH), 1721
2
À1
À1
and 1662 cm (2 C═O), and 1393and 1142 cm (SO2).
Scheme 1
Journal of Heterocyclic Chemistry
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Quinazolines Carrying Sulfonamide Moiety as Anticancer and Radiosensitizing Agents
Scheme 2
Scheme 3
cyclic triazoloquinazoline 12 was obtained (Scheme 3).
The structure of compounds 11 and 12 was established
1
by elemental analysis, IR, H NMR, and mass spectral
data. IR spectrum of compound 11 showed bands at
À1
À1
À1
3
223 cm (NH), 2208 cm (C ꢀ N), 1652 cm (C═O),
À1
1
and 1398 and 1139 cm (SO ). The H NMR spectrum
2
of compound 11 in (DMSO-d ) exhibited signals at
6
4
.1 ppm for (CH ) group and 13.1 ppm because of
2
(
SO NH) group. The mass spectrum of compound 11
2
1
+
The H NMR spectrum of compound 7 in (DMSO-d )
revealed a molecular ion peak m/z at 475 [M ] (0.15%),
6
revealed signals at 2.6 ppm for (COCH ) group and
with a base peak m/z at 373 (100%). The IR spectrum
3
À1
1
1.7 ppm because of (NH) group. The IR spectrum
of compound 12 showed bands at 3356 cm
(NH),
À1
À1
À1
À1
of compound 8 showed bands at 3272 cm
1
(NH),
(SO2).
2206 cm
(C ꢀ N), 1645 cm
(C═O), and 1393 and
À1
À1
1
686 cm
(C═O), and 1359 and 1169 cm
1139 cm (SO ). The H NMR spectrum of compound
2
1
Its H NMR spectrum in (DMSO-d ) revealed signals at
12 in (DMSO-d ) revealed signal at 4.2 ppm corresponding
6
6
1
.2 ppm assigned to (CH ) group.
Reaction of 5b with ethyl bromoacetate in refluxing
to (CH ) group. The mass spectrum of compound 12
revealed a molecular ion peak m/z at 457 [M ] (18.8%),
3
2
+
sodium ethoxide yielded the triazinoquinazoline 9b via the
formation of the intermediate 9a rather than its isomeric
structure 10b (Scheme 2). Structure 9b was suggested rather
than structure 10b, on the basis of the assumption that the
reaction basic condition allowed it to proceed through
formation of sodium salt on the less basic NH, and elimina-
tion of sodium bromide followed by cyclization [45]. The IR
spectrum of the isolated compound 9b showed bands at
with a base peak m/z at 92 (100%). On the other hand,
refluxing compound 5 with carbon disulfide in pyridine
yielded the 2-thioxo-1,2,4-triazolo[5,1-b]quinazoline deriva-
À1
tive 13. Its IR spectrum showed bands at 3223 cm (NH),
À1
À1
À1
1663cm
(C═O), 1391 and 1130cm
(SO ), and
2
1
1250cm (C═S). The H NMR spectrum of compound
13 in (DMSO-d ) revealed signal at 13.1 because of
6
(SO NH) group. The mass spectrum of compound 13
2
À1
À1
+
3
484cm (NH), and 1743 and 1675 cm (2 C═O), which
revealed a molecular ion peak m/z at 450 [M ] (7.0%), with
was at less frequency than that expected for structure 10b.
Further evidence was the H NMR spectrum, which showed
a singlet at 3.9 ppm for the methylene protons.
When compound 5b was reacted with ethyl cyanoace-
tate under condition of fusion, the 3-cyanoacetamido-
quinazoline derivative 11 was obtained in good yield,
whereas in the presence of triethylamine as catalyst, the
a base peak m/z at 344 (100%).
1
The Schiff’s base derivative 14 was obtained via
reaction of compound 5b with benzaldehyde in refluxing
ethanol (Scheme 4). The structure of compound 14 was
1
confirmed by elemental analysis, IR, NMR, and mass
spectral data. The IR spectrum of compound 14 showed
À1
À1
bands at 3319 and 3203 cm (NH), 1665 cm (C═O),
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

M. M. Ghorab, F. A. Ragab, H. I. Heiba, and A. A. Bayomi
Vol 000
Scheme 4
mass spectrum of compound 16 revealed a molecular ion
peak m/z at 509 [M ] (15.6%), with a base peak m/z at 57
+
(100%). Also, compound 16 was obtained by refluxing
compound 15 in pyridine (mp and mixed mp).
In vitro anticancer screening. The results of anticancer
activity (Table 1) indicated that compounds 14 containing
benzylidenamino at position-3 with the biologically active
sulfonamide moiety at position-2 (IC50 value = 22.11 mM)
and 15 having phenylthioureido at position-3 with
sulfonamide moiety at position-2 (IC50 = 19.70 mM) were
found to be potent than all the synthesized compounds
and less active than doxorubicin as reference drug.
Radiosensitizing evaluation. The most potent compounds
resulted from the in vitro anticancer screening; the
quinazolone derivatives 14 and 15 were selected to be
evaluated again for their in vitro anticancer activity alone
and in combination with g-radiation. This study was
conducted to evaluate the ability of these compounds to
enhance the cell killing effect of g-radiation. Cells were
subjected to a single dose of g-radiation at a dose level of
À1
1
and 1394 and 1141 cm (SO ). Its H NMR spectrum in
2
(
DMSO-d ) revealed signal at 8.2 ppm because of (N═CH).
6
The mass spectrum of compound 14 revealed a molecular
+
ion peak m/z at 496 [M ] (0.5%), with a base peak m/z at
4
63 (100%). Our work was extended to study the reactivity
8
Gy with a dose rate of 2 Gy/min. Irradiation was
of compound 5b toward phenyl isothiocyanate. Thus,
reaction of 5b with phenyl isothiocyanate in refluxing ethanol
furnished the corresponding thioureido derivative 15,
whereas in refluxing pyridine, the triazoloquinazoline 16
was obtained. The structure of compounds 15 and 16 was
performed in the National Cancer Institute, Cairo University,
60
using Gamma cell-40 ( Co) source. The surviving fractions
were expressed as means Æ standard error. The results were
analyzed using 1-way ANOVA test and given in Table 2.
1
established by elemental analyses, IR, H NMR, and mass
spectral data. The IR spectrum of compound 15 showed
bands at 3240 and 3203cm (NH). The H NMR spectrum
EXPERIMENTAL
À1
1
Melting points are uncorrected and were determined on a
Stuart melting point apparatus (Stuart Scientific, Redhill, UK).
Elemental analysis (C, H, N) were performed on Perkin-Elmer
2400 analyser (Perkin-Elmer, Norwalk, CT, USA) at the microan-
alytical laboratories of the Faculty of Science, Cairo University.
of compound 15 in (DMSO-d ) revealed signals at 4.5 ppm
6
assigned to (2NH) of thioureido group. The IR spectrum of
À1
compound 16 showed bands at 3463 and 3234 cm (NH),
À1
À1
1
663cm (C═O), and 1356 and 1139 cm (SO ). The
2
Table 1
In vitro cytotoxic activity of the synthesized compounds against human liver cell line (HEPG2).
#
Surviving fraction (means Æ SE)
Compound concentration (mM)
Comp. No.
50
25
12.5
5
IC50
Dox.
4
0.1011 Æ 0.0473
0.3497 Æ 0.0064
0.3536 Æ 0.0181
0.3822 Æ 0.0174
0.4997 Æ 0.0110
0.6051 Æ 0.0245
0.7637 Æ 0.0666
0.3255 Æ 0.0407
0.4675 Æ 0.0285
0.4596 Æ 0.0207
0.4404 Æ 0.1207
0.3182 Æ 0.0156
0.3634 Æ 0.0255
0.0633 Æ 0.0101
0.3114 Æ 0.0164
0.3901 Æ 0.0166
0.4047 Æ 0.0318
0.4843 Æ 0.0340
0.5998 Æ 0.0327
0.9102 Æ 0.0698
0.3544 Æ 0.0123
0.4477 Æ 0.0246
0.4440 Æ 0.0387
0.4886 Æ 0.0965
0.3588 Æ 0.0246
0.2911 Æ 0.0051
0.0615 Æ 0.0116
0.4781 Æ 0.0007
0.6979 Æ 0.0395
0.5438 Æ 0.0580
0.5541 Æ 0.0354
0.8783 Æ 0.0345
0.9473 Æ 0.0346
0.7150 Æ 0.0449
0.6388 Æ 0.0655
0.7931 Æ 0.0812
0.4242 Æ 0.0930
0.3878 Æ 0.0149
0.6162 Æ 0.0437
0.0967 Æ 0.0247
0.9047 Æ 0.0337
0.7773 Æ 0.0680
0.9456 Æ 0.0176
0.8744 Æ 0.0087
0.8567 Æ 0.0630
1.0110 Æ 0.0162
0.8220 Æ 0.0558
0.7573 Æ 0.0208
0.8379 Æ 0.0120
0.2313 Æ 0.0133
0.4880 Æ 0.0178
0.8643 Æ 0.0150
4.15
26.90
30.32
30.84
38.42
55.32
105.63
29.16
35.67
37.63
22.11
19.70
28.30
5
6
7
8
9
1
1
1
1
1
1
b
1
2
3
4
5
6
#
Each value is the mean of three values Æ standard error.
Journal of Heterocyclic Chemistry
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Month 2014
Quinazolines Carrying Sulfonamide Moiety as Anticancer and Radiosensitizing Agents
Table 2
In vitro anticancer screening of compounds 14 and 15 against human liver cell line (HEPG2) in combination with g-radiation.
#
Surviving fraction (means Æ SE)
Irradiated
Control
(8 Gy)
Compound concentration (mM) + irradiation (8 Gy)
Comp. No.
Dox.
5
12.5
25
50
IC50 (mM)
1.000
1.000
1.000
0.927 Æ 0.02*
0.927 Æ 0.02*
0.113 Æ 0.0012*
0.498 Æ 0.0712*
0.4266 Æ 0.04702*
0.53 Æ 0.08505*
0.96 Æ 0.111*
3.67
17.92
15.71
14
5
0.3111 Æ 0.01396* 0.3221 Æ 0.01176* 0.3442 Æ 0.02365*
1
0.927 Æ 0.02* 0.4766 Æ 0.03177* 0.2911 Æ 0.03588* 0.3001 Æ 0.00329* 0.2992 Æ 0.04411*
#
Each value is the mean of three values Æ standard error.
*
Significant difference from control group at p < 0.001.
1
All compounds were within Æ0.4% of the theoretical values. The
IR spectra (KBr) were measured on Shimadzu IR 110 spectropho-
tometer (Shimadzu, Koyoto, Japan), and 1H NMR spectra were
obtained on a Bruker proton NMR-Avance 300 (300 MHz)
(NH), 3045 (CH arom.), 1663 (C═O), 1395, 1139 (SO
NMR (DMSO-d )d: 6.9–7.8 [m,12H,Ar–H], 13.1 [s,1H, SO
O-exchangeable ], 8.0 [s,1H,CH triazole]. MS, m/z (%): 418
2
).
H
6
2
NH,
D
2
+
[M ] (1.65), 88(100). Anal. Calcd for C H N O S: C, 57.41; H,
20 14 6 3
(
Bruker, Munich, Germany), in DMSO-d as a solvent, usingTMS
3.37; N, 20.08. Found: C, 57.17; H 3.15; N, 19.8.
6
as internal standard. Mass spectra were run on HP Model MS-5988
Hewlett Packard, Palo, Alto, California, USA). All reactions
N-(4-oxo-2-(4-(N-pyridin-2-ylsulfamoyl)phenylamino)quinazolin-
(
3(4H)-yl)acetamide (7).
A solution of compound 5b (4.0 g,
were monitored by TLC using precoated Aluminum sheets Silica
gel Merck 60F254 and were visualized by UV lamp (Merck,
Darmstadt, Germany).
0.01 mol) in acetic anhydride (20mL) was refluxed For 10min.
The formed solid mass was collected and crystallized from
ꢁ
ethanol to give compound 7: yield, 68%; mp 149–151 C; IR
À1
Methyl 2-isothiocyanatobenzoate (2).
This was prepared
(KBr, cm ): 3190 (NH), 3065 (CH arom.), 1721, 1662
1
according to the previously reported procedure [43].
(2 C═O), 1393, 1142 (SO
2
). H NMR (DMSO-d
6
)d: 2.6 [s,3H,
Methyl 2-(3-(4-(N-pyridin-2-ylsulfamoyl)phenyl)thioureido)
benzoate (4). A mixture of methyl-2-isothiocyanatobenzoate
COCH ], 7.3–8.2 [m,13H,Ar–H + NH], 11.7 [s,1H, SO NH,
3
2
D O-exchangeable,]. Anal. Calcd for C H N O S C, 55.99; H,
2
21 18 6 4 :
2
(1.9 g, 0.01M) and pyridin-2-yl benzenesulfonamide 3
4.03; N, 18.66. Found: C, 55.97; H, 3.88; N, 18.88.
2-(4-Methyl-9-oxo-[1,2,4]triazolo[5,1-b]quinazolin-3(9H)-yl)-
N-(pyridin-2-yl)benzenesulfonamide (8). solution of
(
2.49 g, 0.01 mol) in dimethylformamide (20 mL) was stirred
at room temperature for 5 h. The reaction mixture was poured
onto cold water, and the obtained solid was crystallized
from dioxane to give compound 4: yield, 94%; mp 235–237 C;
A
compound 5b (4.0 g, 0.01 mol) in acetic anhydride (20 mL) was
refluxed for 10 h. The formed solid mass was collected and
crystallized from ethanol to give compound 8: yield, 66%; mp
ꢁ
À1
IR (KBr, cm ): 3300, 3238 (NH), 3041 (CH arom.), 1651
1
ꢁ
À1
(
C═O), 1262 (C═S), 1389, 1133 (SO
2
). H NMR (DMSO-d
6
)
296–298 C; IR (KBr, cm ): 3272 (NH), 1686 (C═O), 1359,
1
d: 3.6 [s,3H,OCH
3
], 7.0–8.0 [m,14H,Ar–H + 2NH], 10.26
s,1H, SO NH, D2O-exchangeable]. MS, m/z (%): 429
1169 (SO
2 6 3
). H NMR (DMSO-d )d: 1.2 [s,3H,CH ], 7.2–8.2
[
[
[m,12H,Ar–H], 13.4 [s,1H, SO NH, D O-exchangeable ]. Anal.
2
2
2
+
M À CH ] (0.4), 226 (100). Anal. Calcd for C H N O S :
Calcd for C H N O S: C, 58.32; H, 3.73; N, 19.43. Found: C,
21 16 6 3
58.44; H 3.78; N, 19.22.
3
20 18
4
4
2
C, 54.28; H, 4.10; N, 12.66. Found: C, 54.44; H, 3.99;
N, 12.87.
4-(2,10-Dioxo-2,3-dihydro-1H-[1,2,4]triazino[3,2-b]quinazolin-
4(10H)-yl)-N-(pyridin-2-yl)benzenesulfonamide (9). A mixture
of compound 5b (4.0g, 0.01 mol) and ethyl bromoacetate (1.69 g,
0.01 mol) and sodium ethoxide (0.68 g, 0.01mol) in ethanol
(30 mL) was refluxed for 8 h. The reaction mixture was filtered
while hot, and the solid obtained was crystallized from dioxane to
4-(3-Amino-4-oxo-3,4-dihydroquinazolin-2-ylamino)-N-(pyridin-
2
-yl) benzenesulfonamide (5b). A mixture of compound 4
(4.4 g, 0.01 mol) and hydrazine hydrate (1g, 0.02mol) in ethanol
(30mL) was refluxed till evolution of hydrogen sulfide gas was
ceased (lead acetate paper) (14 h.). The reaction mixture was
filtered while hot, and the solid obtained was crystallized from
dioxane to give compound 5: yield, 80%; mp 255–257 C; IR
ꢁ
À1
give compound 9: yield, 78%; mp 182–184 C; IR (KBr, cm ):
ꢁ
3484 (NH) 3065 (CH arom.), 2922, 2846 (CH aliph.), 1743,
À1
1
(KBr, cm ): 3410, 3308, 3183 (NH, NH
2
), 3045 (CH arom.),
). H NMR (DMSO-d )d: 6.8
O-exchangeable], 7.3–8.0 [m,13H,Ar–H + NH],
1675 (2C═O), 1366, 1136 (SO
[s,2H, CH
exchangeable ], 13.2 [s,1H, SO
Calcd for C H N O S: C, 56.24; H, 3.60; N, 18.74. Found: C,
2
). H NMR (DMSO-d
6
)d: 3.9
O-
2
O-exchangeable ]. Anal.
1
1
660 (C═O), 1383 1133 (SO
2
6
2
CO], 7.2–7.9 [m,12H,Ar–H], 8.2 [s,1H, NH, D
NH, D
2
[s,2H,NH2,
D
2
2
+
1
3.2 [s,1H, SO NH, D O-exchangeable ]. MS, m/z (%): 408 [M ]
2 2
21 16 6 4
(
2.1), 343 (100). Anal. Calcd for C H N O S: C, 55.87; H,
55.97; H 3.75; N, 18.55.
1
9 16 6 3
3
.95; N, 20.58. Found: C, 55.97; H, 3.75; N, 20.88.
-(9-Oxo-[1,2,4]triazolo[5,1-b]quinazolin-3(9H)-yl)-N-(pyridin-
-yl) benzenesulfonamide (6). A solution of compound 5b
2-Cyano-N-(4-oxo-2-(4-(N-pyridin-2-ylsulfamoyl)phenylamino)
4
quinazolin-3(4H)-yl)acetamide (11).
A
solution of
2
compound 5b (4.0 g, 0.01 mol) and ethyl cyanoacetate (10 mL)
was refluxed for 8 h. The formed solid mass was collected
and crystallized from methanol to give compound 11: yield,
(4.0 g, 0.01 mol) in formic acid (20 mL) was refluxed for 5h. The
reaction mixture was cooled and then poured onto cold water,
and the obtained solid was crystallized from dioxane to give
compound 6: yield, 71%; mp 281–283 C; IR (KBr, cm ): 3227
ꢁ
À1
84%; mp >300 C; IR (KBr, cm ): 3223 (NH), 2934, 2867
(CH aliph.), 2208 (CN), 1652 (C═O), 1398, 1139 (SO ).
ꢁ
À1
2
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M. M. Ghorab, F. A. Ragab, H. I. Heiba, and A. A. Bayomi
Vol 000
1
H NMR (DMSO-d )d: 4.1 [s,2H, CH CN], 7.4–8.0 [m,14H,
4-(9-oxo-2-(phenylamino)-[1,2,4]triazolo[5,1-b]quinazolin-
3(9H)-yl)-N-(pyridin-2-yl)benzenesulfonamide (16). A mixture
of compound 5b (4.0g, 0.01 mol) and phenyl isothiocyanate
(1.35 g, 0.01 mol) in pyridine (30mL) was refluxed for 12h. The
reaction mixture was poured onto ice/water acidified by dil HCl,
and the solid obtained was crystallized from dioxane to give
6
2
Ar–H + 2NH], 13.1 [s,1H, SO NH, D O-exchangeable].
2
2
+
MS, m/z (%): 475 [M ] (0.15), 373 (100). Anal. Calcd for
S: C, 55.57; H, 3.60; N, 20.62. Found: C,
5.44; H 3.88; N, 20.42.
-(2-(Cyanomethyl)-9-oxo-[1,2,4]triazolo[5,1-b]quinazolin-
(9H)-yl)-N-(pyridin-2-yl)benzenesulfonamide (12). A mixture
22 17 7 4
C H N O
5
4
ꢁ
À1
3
compound 16: yield, 73%; mp: 298–300 C; IR (KBr, cm ):
of compound 5b (4.0g, 0.01 mol) and ethyl cyanoacetate (10mL)
in dimethylformamide (10 mL) containing three drops of
triethylamine was refluxed for 8 h. The formed solid mass was
collected and crystallized from dioxane to give compound 12:
3463, 3234 (NH), 3045 (CH arom.), 1663 (C═O), 1385, 1139
1
(SO
2
). H NMR (DMSO-d
6
)d: 7.2–8.1[m,18H,Ar–H + NH], 13.1
+
[s,1H,SO NH, D O-exchangeable].MS, m/z (%): 509 [M ] (15.6),
2
2
57 (100). Anal. Calcd for C H N O S C, 61.29; H, 3.76; N,
26 19 7 3 :
ꢁ
À1
yield, 84%; mp >300 C; IR (KBr, cm ): 3356 (NH), 2928,
870 (CH aliph.), 2206 (CN), 1645 (C═O), 1393, 1139
19.24. Found: C, 61.44; H, 3.92; N, 19.50.
2
In vitro anticancer screening. Human tumor liver cell line
1
(
[
SO
2
). H NMR (DMSO-d
6
)d: 4.2 [s,2H,CH
2
CN], 7.3–8.0
(HEPG2) was used in this study. The cytotoxic activity was
measured in vitro for the newly synthesized compounds using
the Sulfo-Rhodamine-B stain assay using the method of
Skehan et al. [46]. The in vitro anticancer screening was
performed by the pharmacology unit at the National Cancer
Institute, Cairo University.
m,12H,Ar–H], 13.4 [s,1H, SO NH, D O-exchangeable ].MS,
2
2
+
m/z (%): 457 [M ] (18.8), 92 (100). Anal. Calcd for
S: C, 57.76; H, 3.30; N, 21.43. Found: C, 57.97;
H, 3.65; N, 21.22.
-(2-Sulfanylidine-9-oxo-[1,2,4]triazolo[5,1-b]quinazolin-3(9H)-
yl)-N-(pyridin-2-yl)benzenesulfonamide (13). A solution of
compound 5b (4.0g, 0.01 mol) in carbondisulfide (1.52 g,
.02mol) in pyridine (20mL) was refluxed for 10 h. The reaction
22 15 7 3
C H N O
4
4
Cells were plated in 96-multiwell plate (10 cells/well) for 24 h
before treatment with the compound(s) to allow attachment of cell
to the wall of the plate. Test compounds were dissolved in
dimethyl sulfoxide. Different concentrations of the compound
under test (5, 12.5, 25, and 50 mM) were added to the cell mono-
layer. Triplicate wells were prepared for each individual concen-
tration. Monolayer cells were incubated with the compound(s)
0
mixture was cooled and then poured onto cold water, and the
obtained solid was crystallized from acetic acid to give
ꢁ
À1
compound 13: yield, 68%; mp >300 C; IR (KBr, cm ): 3223
(
NH), 3038 (CH arom.), 1663 (C═O), 1203 (C═S), 1391, 1130
1
ꢁ
(
SO
3.1 [s,1H, SO
7.0), 344 (100). Anal. Calcd for C H N O S : C, 53.32; H,
2
),. H NMR (DMSO-d
6
)d: 7.2–8.0 [m,12H,Ar–H + NH],
for 48 h at 37 C and in atmosphere of 5% CO2. After 48 h, cells
+
1
(
2
NH, D O-exchangeable]. MS, m/z (%): 450 [M ]
2
were fixed, washed, and stained for 30 min with 0.4% (wt/vol)
Sulfo-Rhodamine-B stain dissolved in 1% acetic acid. Excess
unbound dye was removed by four washes with 1% acetic acid,
and attached stain was recovered with Tris-EDTA buffer. Color
intensity was measured in an enzyme-linked immunosorbent
assay reader. The relation between surviving fraction and drug
concentration is plotted to obtain the survival curve for liver
tumor (HEPG2) cell line after the specified time. The molar con-
centration required for 50% inhibition of cell viability (IC50) was
calculated and compared with the reference drug doxorubicin
(CAS, 25316-40-9). The surviving fractions were expressed as
means Æ standard error.
20 14 6 3 2
3
.13; N, 18.66 . Found: C, 53.11;H 2.93;N, 18.88.
N-benzyl-4-(3-(benzylideneamino)-4-oxo-3,4-dihydroquinazolin-
-ylamino)benzenesulfonamide (14). A mixture of compound
b (4.0 g, 0.01 mol) and benzaldehyde (1 g, 0.01 mol) in
2
5
ethanol (30 mL) was refluxed for 12 h. The reaction mixture
was filtered while hot, and the solid obtained was crystallized
ꢁ
from dioxane to give compound 14: yield, 72%; mp 223–225 C;
À1
IR (KBr, cm ): 3319, 3203 (NH), 2923, 2845 (CH aliph.),
1
1
665 (C═O), 1394, 1141 (SO
2
). H NMR (DMSO-d
6
)d: 6.9–8.1
[
m, 18H,Ar–H + NH], 8.2 (s, 1H,N═CH), 9.8 [s,1H, SO NH,
2
13
D O-exchangeable]. C-NMR (DMSO-d ): 112.6, 113.0, 115.7
Radiosensitizing evaluation. The rationale for combining
chemotherapy and radiotherapy is based mainly on two ideas,
one being spatial cooperation, which is effective if
chemotherapy is sufficiently active to eradicate subclinical
metastases and if the primary local tumor is effectively
treated by radiotherapy. In this regard, no interaction
between radiotherapy and chemotherapy is required. The
other idea is the enhancement of radiation effects by direct
enhancement of the initial radiation damage by incorporating
drugs into DNA, inhibiting cellular repair, accumulating cells
in a radiosensitive phase or eliminating radioresistant phase
cells, eliminating hypoxic cells, or inhibiting the accelerated
repopulation of tumor cells. Virtually, all chemotherapeutic
agents have the ability to sensitize cancer cells to the lethal
effects of ionizing radiation [47].
2
6
(2), 119.4, 121.6, 126.7, 127.8 (2), 127.9, 129.3 (2), 130.6 (2),
1
30.9, 132.8, 134.5, 135.1, 139.0, 141.9, 145.3, 148.2, 149.7,
+
154.6, 161.3, 165.1. MS, m/z (%): 496[M ] (0.5), 463(100). Anal.
Calcd for C H N O S C, 62.89; H, 4.06; N, 16.39. Found: C,
26 20 6 3 :
62.91; H, 4.1; N, 16.42.
4
-(4-oxo-3-(3-phenylthioureido)-3,4-dihydroquinazolin-2-
ylamino)-N-(pyridin-2-yl)benzenesulfonamide (15). A mixture
of 5b (4.0g, 0.01M) and phenyl isothiocyanate (1.35 g, 0.01 mol)
in ethanol (30 mL) was refluxed for 5 h. The reaction mixture
was filtered while hot, and the solid obtained was crystallized
ꢁ
from ethanol to give compound 15: yield, 51%; mp:76–78 C.
À1
IR (KBr, cm ): 3240, 3203 (NH), 3036 (CH arom.), 1663
C═O), 1236 (C═S), 1320, 1100 (SO2). H NMR (DMSO-
1
(
d )d: 4.5 (s,2H,2NH of thioureido, D O-exchangeable), 7.1–7.7
6
2
[
m, 20H,Ar–H + 3NH], 11.2 (s,1H,SO
2
NH, D
2
O-exchangeable).
Calculation of IC .
The surviving fractions of cells for
50
13
C-NMR (DMSO-d
6
): 110.5, 114.6, 115.2 (2), 119.7, 121.3,
each tested compound and the reference drug in concentrations
(5, 12.5, 25, and 50 mM) were the average of three tests.
Surviving fraction of cells in control test (0 mM) was considered
as 100% viable cells. Concentrations of the tested compounds
and the reference drug were plotted against surviving fractions
of the cells using Microsoft Excel (2007). A trend line was
drawn for each curve, and a corresponding equation was
1
1
1
23.8, 125.6 (2), 128.0, 128.4 (2), 129.6, 130.5 (2), 131.1,
32.7, 138.6, 139.9, 149.0, 149.7, 151.3, 154.2, 163.9, 168.0,
+
79.3. MS, m/z (%): 543[M ] (5.01), 135 (100). Anal. Calcd for
C
26
H
21
N
7
O
3
S
2: C, 57.44; H, 3.89; N, 18.04. Found: C, 57.65;
H, 3.73; N, 17.88.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2014
Quinazolines Carrying Sulfonamide Moiety as Anticancer and Radiosensitizing Agents
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Compound 14
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Compound 15
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1
50
[
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