Synthesis and Anticancer Evaluation of Some Novel Quinazolin-4(3H)-one Derivatives

Article abstract of DOI:10.1002/jhet.3559

A new series of quinazolinone derivatives bearing pyridine, pyrimidine, pyrazole, or pyran moieties were synthesized for the purpose of anticancer cell line evaluation. Synthesis of these derivatives was achieved by the reaction of the ketone 2 with the appropriate aldehydes in the presence of either ethylcyanoacetate or malononitrile and ammonium acetate in one-pot reaction. Chalcones 6 reacted also with hydrazine hydrate to give the corresponding pyrazolines 7 and reacted with urea or thiourea to give the 2-oxopyrimidines or the 2-thioxopyrimidines 8, respectively. Evaluation of some representative examples of the newly synthesized compounds against cancer cell lines showed promising activity as anticancer agents.

Full text of DOI:10.1002/jhet.3559

Month 2019
Synthesis and Anticancer Evaluation of Some Novel Quinazolin-4(3H)-
one Derivatives
a
*
Rasha S. Gouhar,
David S. A. Haneen,b and Salwa M. El-Halloutyc
aTherapeutical Chemistry Department, National Research Centre, Dokki, P. O 12622, Giza, Egypt
bChemistry Department, Faculty of Science, Ain Shams University, Abbassia, P. O 11566, Cairo, Egypt
cDrug Bioassay-Cell Culture Laboratory, Department of Pharmacognosy, National Research Centre, Dokki, P. O 12622,
Giza, Egypt
*E-mail: dr.rasha_gouhar@yahoo.com
Received September 30, 2018
DOI 10.1002/jhet.3559
Published online 00 Month 2019 in Wiley Online Library (wileyonlinelibrary.com).
A new series of quinazolinone derivatives bearing pyridine, pyrimidine, pyrazole, or pyran moieties were
synthesized for the purpose of anticancer cell line evaluation. Synthesis of these derivatives was achieved by
the reaction of the ketone 2 with the appropriate aldehydes in the presence of either ethylcyanoacetate or
malononitrile and ammonium acetate in one-pot reaction. Chalcones 6 reacted also with hydrazine hydrate
to give the corresponding pyrazolines 7 and reacted with urea or thiourea to give the 2-oxopyrimidines or
the 2-thioxopyrimidines 8, respectively. Evaluation of some representative examples of the newly synthe-
sized compounds against cancer cell lines showed promising activity as anticancer agents.
J. Heterocyclic Chem., 00, 00 (2019).
INTRODUCTION
over expression through the inhibition of EGFR
autophosphorylation such as Gebitinib (III) [13] and
Erlotinib (IV) (Fig. 1) [14].
On the other hand, it has been reported that several
pyridines, pyrans, pyrazolines, and pyrimidin-2-ones or
pyrimidine-2-thiones besides α, β-unsaturated ketones
found to possess considerable wide range of
pharmacological activities especially as anticancer agents
[15–18]. The aim of the present investigation is to
It has been obvious that quinazolines and quinazoline-
4(3H)-one derivatives have gained extensive interest due
to their wide range of pharmaceutical and biological
activities [1–7], they are being extensively utilized as
drug-like scaffold in medicinal chemistry [8] especially as
anticancer agents through various mechanisms. Some of
quinazoline derivatives are considered as antifolate
thymidylate synthase inhibitors such as Nolatrexed (I)
synthesize
a
new series of quinazoline-4(3H)-one
[9],
while
2-substituted-1,3-dihydro-quinazolinone
derivatives incorporated into the above-mentioned
heterocyclic moieties at position 3 of the quinazoline
system. The presence of these heterocyclic rings in one
molecule may results in a dual mode of biological action,
which may increase the promising anticancer activity
against cancer cell lines.
derivatives are associated with inhibitory effects on
tubulin polymerization such as GMC-5-193 (II) [10,11].
Also, some 4-anilinoquinazoline represents a new class of
antitumor drugs [12]. They were found to inhibit the
epidermal growth factor receptor [EGFR] tyrosine kinase
© 2019 Wiley Periodicals, Inc.

R. S. Gouhar, D. S. A. Haneen, and S. M. El-Hallouty
Vol 000
Figure 1. Structures of compounds I–IV.
RESULTS AND DISCUSSION
piperidine gave the corresponding 2-aminopyran, namely,
2-amino-6-(4-(2-(4-chlorophenyl)-4-oxoquinazolin-3(4H)-
yl)phenyl)-4-(aryl-2-yl)-4H-pyran-3-carbonitrile (5a–d),
respectively (Scheme 1).
The mechanistic pathway for the conversion of the
ketone 2 into compounds 3a–d, 4a–d, and 5a–d was
represented in (Scheme 2) through the one-pot Michael
addition reaction.
On the other hand, since chalcones (α, β-unsaturated
ketones) are useful starting materials for the synthesis of
several heterocyclic systems such as pyrazolines,
pyridones, and pyrimidine-2-ones or 2-thiones, it was of
our interest to synthesize another new series of these
heterocyclic moieties incorporated to a quinazoline-4(3H)-
one system through a p-phenyl group for the purpose of
anticancer evaluation. Claisen-Schmidt condensation of
the acetyl derivatives 2 with the appropriate aldehydes in
5–10% ethanolic sodium hydroxide solution afforded the
corresponding 1,3-propen-1-ones 6 in good yield. The
chalcones 6, namely, 2-(4-chlorophenyl)-3-(4-(3-(aryl/
Chemistry. The chemical modification of 3-substituted
quiazoline-4(3H)-one derivatives such as the introduction
of different substituents or heterocyclic ring systems
allowed an expansion of SAR studies to afford new
insight into molecular interaction at receptor level in the
cancer cell. Recently, it is well established in our drug
discovery program [19,20] that slight structural
modification on the substitution at positions 2 and 3 of
the quiazoline-4(3H)-one ring system increased the
anticancer activity against human breast carcinoma cell
lines (MCF-7) through inhibition of PARP-1 enzyme
[poly (ADP-ribose)polymerase-1-enzyme] [19,20].
In the present work, this chemical modification was
achieved by a method one-step synthesis of some new
cyanopyridones, cyanoiminopyridines, and aminopyrans
incorporated to the position 3 of the quinazoline-4(3H)-
one moiety through a para phenyl group. The starting
material 4H-benzo[d][3,1]oxazin-4-one derivative (1) was
obtained by allowing anthranilic acid dissolved in
pyridine to react with p-chlorobenzoyl chloride at room
temperature. The mass spectrum of 1 shows the correct
molecular ion peak at 257 and 259 corresponding to M
and M + 2, respectively. It is well known that the most
common method to obtain substituted 3H-quiazolin-4-one
derivatives is based on the aminolysis of the
corresponding benzoxazin-4-ones [21]. The synthesis of
the starting compound, namely, 3-(4-acetylphenyl)-2-(4-
chlorophenyl)quinazolin-4(3H)-one (2), was achieved by
fusion of 2-(4-chlorophenyl)-4H-benzo[d][3,1]oxazin-4-
one (1) with p-aminoacetophenone. The one-pot
Michael addition reaction [22] of the ketone 2 with
the appropriate aromatic aldehydes, namely, 3,4-dime-
thoxybenzaldehyde,2,5-dimethoxybenzaldehyde indole-3-
carboxaldehyde and/or naphthalene-2-carboxaldehyde
with ethyl cyanoacetate in the presence of anhydrous
ammonium acetate, afforded the corresponding pyridine-
2(1H)-ones (3a–d), respectively.
indolyl)acryloyl)phenyl)quinazolin-4(3H)-one
(6a–d),
respectively, were allowed to react with hydrazine hydrate
98% to give the corresponding pyrazolines 7, namely,
2-(4-chlorophenyl)-3-(4-(5-(aryl)-4,5-dihydro-1H-pyrazol-
3-yl)phenyl)quinazolin-4(3H)-one (7a,b), respectively.
When the reaction carried out in acetic acid, the N-
acetylpyrazolines 7c,d, namely, 2-(4-chlorophenyl)-3-(4-
(5-(indol-3-yl/naphthalene-3-yl)-4,5-dihydro-1H-pyrazol-3
-yl)phenyl)quina-zolin-4(3H)-ones (7c,d), respectively,
were obtained (Scheme 3). Also, cyclocondensation of the
chalcones 6a,b with urea in the presence of hydrochloric
acid afforded the corresponding pyrimidones, namely,
2-(4-chlorophenyl)-3-(4-(6-(aryl)-2-oxo-1,2-dihydropyri-
midin-4-yl)phen-yl)quinazolin-4(3H)-ones
(8a,b),
respectively, while when chalcones 6c,d were allowed to
react with thiourea in the presence of sodium hydroxide
afforded the corresponding pyrimidinethiones, namely,
2-(4-chlorophenyl)-3-(4-(6-(indol-3-yl/naphthalen-2-yl)-2-
thioxo-1,2-dihydropyrimidin-4-yl)phenyl)
quinazolin-
4(3H)-ones (8c,d), respectively, according to reported
method [16] (Scheme 3).
In the same manner, as a chemical investigation of the
one-pot reaction to obtain the 2(H)-pyridin-carbonitriles
3. The reaction of α, β-unsaturated ketones 6a–d with
ethyl cyanoacetate in the presence of ammonium acetate
In the same manner, the one-pot reaction of the ketone 2
with the same aldehydes and malononitrile in the presence
of anhydrous ammonium acetate afforded the
corresponding 2(1H)-iminopyridine-3-carbonitriles 4a–d,
respectively, while the same one-pot reaction of 2 with
the same aromatic aldehydes and malononitrile in
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2019
Synthesis and Anticancer Evaluation of Some Novel Quinazolin-4(3H)-one
Derivatives
Scheme 1. Formation of compounds 3–5.
All new compounds were illustrated by chemical
Scheme 2. The mechanistic pathway for the formation of compounds 3–5.
1
microanalyses and spectral analyses, IR, H-NMR, 13C-
NMR, and mass spectra.
Antitumor evaluation.
This study exhibited the
antitumor activity of some of the newly synthesized
compounds [23–26]. Fifteen derivatives were selected as
representative examples to examine their in vitro
antitumor activities against HepG2 human liver cancer
cell line and MCF-7 human breast cancer cell line. The
percentage of the intact cells was measured and tabulated
in (Table 1 and Fig. 2). The activities of the derivatives
against carcinoma cells were compared with the
cytotoxicity of doxorubicin as a reference drug. The
results showed that 2-imino-(1H-indol)-4-oxoquinazolin
derivative (4C) exhibited the highest cytotoxic activity
against the liver cancer cells (HepG2) with inhibition
percentage of (99.3%), while they showed moderate
potency against the breast carcinoma cell lines MCF-7
(growth inhibition 76.3%). Fortunately, dual inhibition
potency against both HepG2 and MCF-7 cancer cell lines
was
gained
by
the
(2,5-dimethoxyphenyl)-4-
oxoquinazolin derivative (3b).
The results indicate that (1H-indol)-dihydropyridine-4-
oxoquinazoline derivative (3C) exhibited the highest
cytotoxic activity against breast carcinoma cell lines
(MCF-7) with inhibition percentage of growth inhibition
(92.6%) with lack of activity against (HepG2) cell lines.
The results indicated that the starting (4-acetyl phenyl)-
qunazolin-4-one derivative (2) showed highest potency
cytotoxic activity against the MCF-7 cancer cell lines
in butanol afforded the pyridones 3a–d, respectively, with
the same melting points and mixed melting points with that
obtained by the one-pot reaction, but with lower overall
yield (50%, one-pot reaction yield is 70%) (Scheme 3).
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

R. S. Gouhar, D. S. A. Haneen, and S. M. El-Hallouty
Vol 000
Scheme 3. Formation of compounds 6–8.
inhibition growth, (47.5%, 62.2%, and 65.5%) with lack
of activity against breast carcinoma cell lines MCF-7.
Also, (2,5-dimethoxyphenyl)-2-imino-4-oxoquinazoline
derivative (4b) exhibits moderate activity against liver
carcinoma cell lines (HePG2) with lack activity against
breast carcinoma cell lines MCF-7, while (2,5-
Table 1
% Inhibition of tested compounds at 25 μmol for 48 h.
% Inhibition
No.
Compounds
MCF-7
100
HePG2
100
DOX
dimethoxyphenyl)-
1H-pyrazol-quinazoline-4-one
1
2
3
4
5
6
7
8
2
88.7
0
51.5
47.5
94.6
0
derivative (7b) showed activity against breast carcinoma
cell lines MCF-7 with weak activity against liver
carcinoma cell lines (HepG2). The rest of the tested
derivatives produced weak activity or no activity.
3a
3b
3c
4b
4c
4d
5b
5c
6c
6d
7b
7c
8a
8d
97.5
92.6
20.4
76.3
6.2
17.01
25.3
0
53.5
99.3
62.2
16.5
14.5
11.2
18.6
26.3
35.6
23.7
65.5
9
EXPERIMENTAL
10
11
12
13
14
15
0
All melting points are uncorrected and were taken on
open capillary tubes using electrothermal apparatus 9100.
Elemental micro analyses were carried out at
microanalytical unit, Central Services Laboratory,
National Research Centre, Dokki, Giza, Egypt, using Vario
Elementar and were found within + or À0.5% of the
theoretical values. Infrared spectra were recorded on a
Jasco FTIR-6100, Fourier Transform Infrared Spectrome-
ter at cmÀ1 scale using KBr disc technique at the Central
40.4
0
0
0
(inhibition growth; 88.7%) with moderate activity against
HepG2 cell lines (51.5%).
The (3,4-dimethoxyphenyl)-4-oxoqunazoline derivative
(3a), 2-imino-2-napthalen-4-oxoquinaz-oline derivative
(4d), and 4-naphthalen-2 thioxo-dihydro pyrimidine
quinazoline-4-one derivative (8d) showed moderate
activity against liver carcinoma cell lines (HePG2) with
1
Services Lab. NRC, Dokki, Giza, Egypt. H-NMR and
13C-NMR spectra were determined by using a JEOL
EX-NMR Spectrometer – 300 and 75 MHz – at Central
Services Lab, (Faculty of Science, Cairo University, Cairo,
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2019
Synthesis and Anticancer Evaluation of Some Novel Quinazolin-4(3H)-one
Derivatives
Figure 2. Comparison of the effect of the selected compounds on HepG2 and MCF-7 at 25 μmol for 48 h.
Egypt). Mass spectra were measured with Finnigan M A T
SSQ-7000 mass spectrometer at the Central Services, NRC
Dokki, Giza, Egypt. Follow-up of the reactions and
checking the purity of the compounds were made by
thin-layer chromatography on silica gel-precoated alumi-
num sheets (Type 60 F254-Merck, Darmstadt, Germany),
and the spots were detected by exposure to UV Lamp at
254 nanometer for few seconds. Nomenclature given for
the new compounds is according to the IUPAC System.
2-(4-Chlorophenyl)-4H-benzo[d][1,3]oxazin-4-one (1).
To a solution of anthranilic acid (1.4 g; 0.11 mol)
dissolved in pyridine (60 mL), p-chlorobenzoylchloride
(3.5 g; 2.6 mL; 0.02 mol) was added. The mixture was
stirred for 40 min at room temperature. This was poured
into a beaker containing 5% sodium bicarbonate (50 mL)
solution with stirring. The obtained solid was filtered from
ethanol. Purification of the compound was checked by
thin-layer chromatography using benzene and ethyl acetate
as mobile phase in the ratio of 7:3. I2 vapor was used as
developer. Yield was found to be 82% of brown crystals.
Yellow crystals; yield 80%; m.p. 152–153°C as
reported; IR (KBr, cmÀ1): 1756 (C=O, lactone), 1657
(C=N), 749 (C-Cl); 1H-NMR (300 MHz, DMSO-d6, δ
ppm): 7.30–8.13 (m, 8H, Ar-H). 13C-NMR (75 MHz,
DMSO-d6) δ/ppm: 116.4, 124.9, 126.9, 127.2, 128.3,
128.8, 129.5, 135.8, 136.4, 146.2, 156.1 (Ar-C), 159.3
(C=O). MS (m/z, (relative abundance, %)): 259 (M + 2,
16), 257 (M+, 45), 146 (67), 103 (52), 77 (100), 56 (34).
Anal. Calcd for C14H8ClNO2: C, 65.26; H, 3.13; N, 5.44;
Found: C, 65.30; H, 3.18; N, 5.40.
After cooling, the crude mass was crystallized from
ethanol to give brown crystals of compound 2 in yield 80%.
Pale yellow crystals; yield 63%; m.p. 176–178°C; IR
(KBr, cmÀ1): 1738 (C=O, ketone), 1692 (C=O, cyclic
1
amide), 1652 (C=N), 753 (C-Cl). H-NMR (DMSO-d6,
δ/ppm): 2.90 (s, 3H, CH3), 7.90–8.33 (m, 12H, Ar-H).
13C-NMR (75 MHz, DMSO-d6) δ/ppm: 26.6(CH3),
120.9, 124.5, 126.3, 126.7, 126.9, 127.3, 128.4, 129.3,
129.8, 133.3, 135.7, 136.8, 137.2, 148.7, 156.7 (C=C and
C=N), 160.9 (C=O cyclic ester), 197.3 (C=O). MS (m/z,
(relative abundance, %)): 376 (11), 374 (M+, 35); 273
(21), 177 (32), 151 (53), 69 (32), 57 (42), 55 (100). Anal.
Calcd for C22H15ClN2O2: C, 70.50; H, 4.03; N, 7.47;
Found: C, 70.55; H, 4.13; N, 7.50.
General
procedures
of
synthesis
of
2-oxo-pyridine-3-carbonitrile derivatives 3a–d. Method
(I). A mixture of the ketone 2 (073 g; 0.002 mol), ethyl
cyanoacetate (0.23 mL; 0.002 mol), and the appropriate
aldehydes, namely, 3,4-dimethoxybenzaldehyde, 2,5-
dimethoxybenzaldehyde, indole-3-carboxaldehyde, and
naphthalene-2-carboxaldehyde (0.002 mol) in 10 mL n-
butanol was refluxed for 6 h. The reaction mixture was
concentrated to half of its volume under reduced pressure.
After cooling, the formed precipitate was filtered off,
dried, and recrystallized from the proper solvent to give
compounds 3a–d, respectively, in about 70% yield.
Method (II).
A mixture of the chalcone 6a–d
(0.002 mol), ethyl cyanoacetate (0.23 mL; 0.002 mol),
and anhydrous ammonium acetate (1.24 g; 0.016 mol) in
10 mL n-butanol was refluxed at 150°C for 4 h to give
compounds 3a–d, respectively, in about 50% yield (M.p,
mix M.p.). This indicates that one-pot reaction gives
better yield (about 70%) of the pyridones carbonitriles
than for those obtained from chalcones.
3-(4-Acetylphenyl)-2-(4-chlorophenyl)quinazolin-4(3H)-
one (2). A mixture of compound 1 (2.58 g, 0.01 mol) and
p-aminoacetophenone (1.35 g, 0.01 mol) was heated
together upon fusion at 150–155°C on sand bath for 3 h.
Journal of Heterocyclic Chemistry
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R. S. Gouhar, D. S. A. Haneen, and S. M. El-Hallouty
Vol 000
6-(4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)phenyl)-
4-(3,4-dimethoxyphenyl)-2-oxo-1,2-dihydropyridine-3-
(31), 55 (56). Anal. Calcd for C36H21ClN4O2: C, 74.93;
H, 3.67; N, 9.71; Found: C, 74.95; H, 3.70; N, 9.69.
carbonitrile (3a). Brownish yellow crystals; yield 71%;
General Procedures of Synthesis of Compounds 4a–d.
A
m.p. 115–116°C; IR (KBr, cmÀ1): 3450 (NH), 2229
mixture of the ketone 2 (073 g; 0.002 mol), malononitrile
(0.12 mL; 0.002 mol), anhydrous ammonium acetate
(1.24 g; 0.016 mol), and the appropriate aldehydes,
(CN), 1708, 1685 (2C=O, cyclic amide), 1652 (C=N),
1
1266 (OMe), 744 (C-Cl). H-NMR (DMSO-d6, δ/ppm):
3.30 (s, 3H, OCH3), 4.20 (s, 3H, OCH3), 6.50 (s, 1H,
CH, pyridine), 6.61–8.30 (m, 15H, Ar-H), 11.23 (br.s,
1H, NH, exchangeable). 13C-NMR (75 MHz, DMSO-d6)
δ/ppm: 56.5, 57.3 (2OCH3), 105.6, 111.1, 112.9, 115.3,
115.7, 120.5, 124.3, 124.8, 125.7, 126.4, 126.6, 126.9,
127.3, 127.6, 128.7, 129.3, 131.9, 133.4, 135.7, 136.4,
137.1, 148.7, 149.1, 149.7, 157.4, 159.4 (C=C and C=N),
160.7, 161.9 (2C=O). MS (m/z, (relative abundance, %)):
589 (9), 587 (M+, 28), 553 (17), 239 (25), 105 (67), 77
(74), 44 (100). Anal. Calcd for C34H23ClN4O4: C, 69.57;
H, 3.95; N, 9.54;; Found: C, 69.60; H, 3.90; N, 9.50.
namely,
3,4-dimethoxybenzaldehyde,
2,5-
dimethoxybenzaldehyde, indole-3-carboxaldehyde, and
naphthalene-2-carboxaldehyde (0.002 mol) in 10 mL n-
butanol was refluxed for 5 h. After cooling, the formed
precipitate was filtered off, dried, and recrystallized
from the proper solvent to give compounds 4a–d,
respectively.
6-(4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)phenyl)-
4-(3,4-dimethoxyphenyl)-2-imino-1,2-dihydropyridine-3-
carbonitrile (4a). Brown crystals; yield 53%; m.p. 110–
112°C; IR (KBr, cmÀ1): 3405, 3398 (NH), 2226 (CN),
1678 (C=O, cyclic amide), 1653 (C=N), 1219 (OMe),
6-(4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)phenyl)-
4-(2,5-dimethoxyphenyl)-2-oxo-1,2-dihydropyridine-3-
1
752 (C-Cl). H-NMR (DMSO-d6, δ/ppm): 3.90 (s, 3H,
carbonitrile (3b). Brownish yellow crystals; yield 74%;
OCH3), 4.20 (s, 3H, OCH3), 5.87 (br.s, 1H, NH,
exchangeable), 6.61–8.10 (m, 16H, Ar-H), 9.31 (br.s, 1H,
NH-Imino, exchangeable). MS (m/z, (relative abundance,
%)): 588 (6), 586 (M+, 21), 303 (100), 197 (32), 125
(27). Anal. Calcd for C34H24ClN5O3: C, 69.68; H, 4.13;
m.p. 130–132°C; IR (KBr, cmÀ1): 3450 (NH), 2229
(CN), 1703, 1685 (2C=O, cyclic amide), 1650 (C=N),
1
1266 (OMe), 744 (C-Cl). H-NMR (DMSO-d6, δ/ppm):
3.50 (s, 3H, OCH3), 3.90 (s, 3H, OCH3), 6.50 (s, 1H,
CH, pyridine), 6.61–8.30 (m, 15H, Ar-H), 11.21 (br.s,
1H, NH, exchangeable). MS (m/z, (relative abundance,
%)): 589 (12), 587 (M+, 34), 105(54), 77(100), 56(87).
Anal. Calcd for C34H23ClN4O4: C, 69.57; H, 3.95; N,
9.54;; Found: C, 69.59; H, 3.92; N, 9.51.
N, 11.95; Found: C, 69.70; H, 4.15; N, 11.93.
6-(4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)phenyl)-
4-(2,5-dimethoxyphenyl)-2-imino-1,2-dihydropyridine-3-
carbonitrile (4b). Yellowish white crystals; yield 68%; m.
p. 129–131°C; IR (KBr, cmÀ1): 3405, 3398 (NH), 2226
(CN), 1691 (C=O, cyclic amide), 1656 (C=N), 1219
(OMe), 752 (C-Cl). H-NMR (DMSO-d6, δ/ppm): 3.50
6-(4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)phenyl)-
4-(1H-indol-3-yl)-2-oxo-1,2-dihydropyridine-3-carbonitrile
1
(3c).
Brownish green crystals; yield 57%; m.p. 105–
(s, 3H, OCH3), 3.80 (s, 3H, OCH3), 5.90 (br.s, 1H, NH,
exchangeable), 6.61–8.10 (m, 16H, Ar-H), 9.42 (br.s, 1H,
NH-imino, exchangeable). 13C-NMR (75 MHz,
DMSO-d6) δ/ppm: 55.9, 57.7 (2OCH3), 110.6, 111.4,
112.9, 113.5, 115.6, 120.5, 123.3, 124.3, 124.7, 125.7,
126.4, 126.9, 127.3, 127.7, 128.7, 129.3, 131.9, 133.4,
134.2, 135.7, 136.4, 137.1, 148.7, 149.2, 149.7, 156.1,
158.7 (C=C and C=N), 160.2 (C=O). MS (m/z, (relative
abundance, %)): 588 (8), 586 (M+, 25), 453 (26), 303
(100), 196 (19), 105 (73). Anal. Calcd for
C34H24ClN5O3: C, 69.68; H, 4.13; N, 11.95; Found: C,
107°C; IR (KBr, cmÀ1): 3470, 3458 (NH), 2261 (CN),
1715, 1688 (2C=O, cyclic amide), 1655 (C=N), 742 (C-
Cl). 1H-NMR (DMSO-d6, δ/ppm): 6.50 (s, 1H, CH,
pyridine), 7.30–7.61 (m, 17H, Ar-H), 11.23 (br.s, 1H,
NH, exchangeable), 11.89 (br.s, 1H, NH, exchangeable).
13C-NMR (75 MHz, DMSO-d6) δ/ppm: 105.3, 111.1,
112.6, 115.5, 115.7, 120.5, 121.3, 123.1, 124.7, 124.1,
125.7, 126.4, 126.7, 126.9, 127.3, 127.6, 128.7, 129.3,
131.7, 133.4, 135.1, 136.6, 137.1, 148.7, 149.1, 149.69,
157.3, 159.4 (C=C and C=N), 160.5, 161.9 (2C=O). MS
(m/z, (relative abundance, %)): 568 (8), 566 (M+, 27),
333 (100), 302 (35), 246 (57), 151 (78), 115 (24), 91
(24), 77 (32). Anal. Calcd for C34H20ClN5O2: C, 72.15;
H, 3.56; N, 12.37; Found: C, 72.20; H, 3.60; N, 12.35.
69.73; H, 4.11; N, 11.98.
6-(4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)phenyl)-
2-imino-4-(1H-indol-2-yl)-1,2-dihydropyridine-3-carbonitrile
(4c). Yellow green crystals; yield 67%; m.p. 137–139°C;
IR (KBr, cmÀ1): 3445, 3426, 3388 (NH), 2271 (CN), 1689
6-(4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)phenyl)-
4-(naphthalen-2-yl)-2-oxo-1,2-dihydropyridine-3-carbonitrile
(3d). Yellow crystals; yield 67%; m.p. 144–146°C; IR
1
(C=O, cyclic amide), 1653 (C=N), 738 (C-Cl). H-NMR
(DMSO-d6, δ/ppm): 6.28 (br.s, 1H, NH, exchangeable),
7.30–7.91 (m, 18H, Ar-H), 9.35 (br.s, 1H, NH-imino,
(KBr, cmÀ1): 3405, 3398 (NH), 2217 (CN), 1718, 1698
1
(2C=O, cyclic amide), 1648 (C=N), 755 (C-Cl). H-NMR
exchangeable),
11.93
(br.s,
1H,
NH-indolyl,
(DMSO-d6, δ/ppm): 6.80 (s, 1H, CH, pyridine), 7.30–
7.61 (m, 19H, Ar-H), 11.13 (br.s, 1H, NH,
exchangeable). MS (m/z, (relative abundance, %)): 579
(5), 577 (M+, 15), 357 (18), 303 (23), 151 (100), 121
exchangeable). MS (m/z, (relative abundance, %)): 567
(11), 565 (M+, 31), 464 (43), 303 (100), 91 (76), 77 (45).
Anal. Calcd for C34H21ClN6O: C, 72.27; H, 3.75; N,
14.87; Found: C, 72.30; H, 3.73; N, 14.82.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2019
Synthesis and Anticancer Evaluation of Some Novel Quinazolin-4(3H)-one
Derivatives
6-(4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)phenyl)-
2-imino-4-(naphthalen-2-yl)-1,2-dihydropyridine-3-carbonitrile
91 (53), 77 (41). Anal. Calcd for C34H25ClN4O4: C, 69.33;
H, 4.28; N, 9.51; Found: C, 69.29; H, 4.31; N, 9.45.
2-Amino-6-(4-(2-(4-chlorophenyl)-4-oxoquinazolin-3(4H)-
(4d).
Brown crystals; yield 56%; m.p. 118–120°C; IR
(KBr, cmÀ1): 3452, 3368 (NH), 2258 (CN), 1689 (C=O,
cyclic amide), 1649 (C=N), 749 (C-Cl). 1H-NMR
(DMSO-d6, δ/ppm): 6.13 (br.s, 1H, NH, exchangeable),
7.50–7.81 (m, 20H, Ar-H), 9.18 (br.s, 1H, NH,
exchangeable). 13C-NMR (75 MHz, DMSO-d6) δ/ppm:
111.1, 112.7, 113.5, 114.7, 115.6, 116.3, 117.1, 119.7,
120.2, 123.6, 124.3, 124.9, 125.7, 126.0, 126.4, 126.9,
127.3, 127.7, 128.7, 129.3, 131.9, 133.4, 134.2, 135.7,
136.4, 137.1, 148.7, 149.2, 149.7, 156.1, 158.7 (C=C and
C=N), 160.2 (C=O). MS (m/z, (relative abundance, %)):
578 (12), 576 (M+, 35), 475 (73), 314 (25), 303 (100), 197
(32), 77 (93). Anal. Calcd for C36H22ClN5O: C, 75.06; H,
3.85; N, 12.16; Found: C, 75.10; H, 3.83; N, 12.20.
yl)phenyl)-4-(1H-indol-2-yl)-4H-pyran-3-carbonitrile (5c).
Yellowish green crystals; yield 62%; m.p. 134–136°C; IR
(KBr, cmÀ1): 3443, 3401, 3364 (NH, NH2), 2271 (CN),
1
1688 (C=O, cyclic amide), 1650 (C=N), 745 (C-Cl). H-
NMR (DMSO-d6, δ/ppm): 3.97 (s, 1H, CH-Ar), 4.50 (s,
1H, CH-pyran), 6.80 (br.s, 2H, NH2, exchangeable),
7.30–7.61 (m, 17H, Ar-H), 11.03 (br.s, 1H, NH,
exchangeable). MS (m/z, (relative abundance, %)): 570
(14), 568 (M+, 40), 502 (32), 401 (75), 133 (65), 109
(53), 77 (100). Anal. Calcd for C34H22ClN5O2: C, 71.89;
H, 3.90; N, 12.33; Found: C, 71.90; H, 3.94; N, 12.35.
2-Amino-6-(4-(2-(4-chlorophenyl)-4-oxoquinazolin-3(4H)-
yl)phenyl)-4-(naphthalen-2-yl)-4H-pyran-3-carbonitrile (5d).
General procedures of synthesis of compounds 5a–d.
mixture of the ketone 2 (073 g; 0.002 mol), malononitrile
(0.15 mL; 0.0025 mol), and the appropriate aldehydes,
A
Brownish red crystals; yield 62%; m.p. 145–146°C; IR
(KBr, cmÀ1): 3405, 3398 (NH2), 2217 (CN), 1698 (C=O,
cyclic amide), 1652 (C=N), 755 (C-Cl). 1H-NMR
(DMSO-d6, δ/ppm): 3.95 (s, 1H, CH-Ar), 5.02 (s, 1H, CH-
pyran), 6.80 (br.s, 2H, NH2, exchangeable), 7.30–7.61 (m,
19H, Ar-H). 13C-NMR (75 MHz, DMSO-d6) δ/ppm:
111,8, 112.6, 112.9, 113.8, 114.5, 117.9, 118.1, 120.1,
121.7, 123.6, 125.8, 126.7, 128.3, 130,2, 132.1, 134.7,
135.8, 136.4, 137.1, 137.4, 138.9, 139.6, 140.3, 142.6,
142.7, 143.8, 144.4, 147.4, 149.1, 154.0, 156.4 (C=C and
C=N), 160.9 (C=O). MS (m/z, (relative abundance, %)):
581 (5), 579 (M+, 15), 513 (35), 412 (62), 133 (58), 77
(100). Anal. Calcd for C36H23ClN4O2: C, 74.67; H, 4.00;
N, 9.68; Found: C, 74.65; H, 3.90; N, 9.69.
namely,
3,4-dimethoxybenzaldehyde,
2,5-
dimethoxybenzaldehyde, indole-3-carboxaldehyde, and
naphthalene-2-carboxaldehyde (0.002 mol) in the
presence of few drops of piperidine was refluxed for 5 h.
The reaction mixture was concentrated to half of its
volume under reduced pressure. After cooling, the formed
precipitate was filtered off, dried, and recrystallized from
the proper solvent to give compounds 5a–d, respectively.
2-Amino-6-(4-(2-(4-chlorophenyl)-4-oxoquinazolin-3(4H)-
yl)phenyl)-4-(3,4-dimethoxyphen-yl)-4H-pyran-3-carbonitrile
(5a). Dark orange crystals; yield 62%; m.p. 188–190°C;
IR (KBr, cmÀ1): 3353, 3442 (NH2), 2219 (CN), 1689
(C=O, cyclic amide), 1655 (C=N), 1281 (OMe), 744 (C-
General procedures of synthesis of compounds 6a–d.
A
1
mixture of the ketone 2 (073 g; 0.002 mol), and the
appropriate aldehydes, namely, 3,4-
2,5-dimethoxybenzaldehyde,
and naphthalene -2-
Cl). H-NMR (DMSO-d6, δ/ppm): 3.70 (s, 3H, OCH3),
3.90 (s, 3H, OCH3), 3.94 (d, 1H, CH-Ar, J = 7.5 Hz),
4.92 (d, 1H, CH, pyran, J = 7.3 Hz), 5.97 (br.s, 2H, NH2,
exchangeable), 6.51–7.80 (m, 15H, Ar-H). 13C-NMR
(75 MHz, DMSO-d6) δ/ppm: 56.1, 56.7 (2OCH3), 107.1,
107.8, 111,3, 112.4, 112.9, 113.8, 114.5, 120.1, 121.7,
123.6, 125.1, 126.7, 128.0, 131,2, 133.5, 134.7, 135.8,
137.1, 138.9, 140.3, 142.3, 142.7, 144.4, 147.4, 149.1,
154.2, 156.9 (C=C and C=N), 161.7 (C=O). MS (m/z,
(relative abundance, %)): 591 (15), 589 (M+, 45), 523
(61), 422 (34), 133 (100), 91 (68), 77 (73). Anal. Calcd
for C34H25ClN4O4: C, 69.33; H, 4.28; N, 9.51; Found: C,
dimethoxybenzaldehyde,
indole-3-carboxaldehyde,
carboxaldehyde (0.002 mol) in 10 mL ethanol, 5% NaOH
in ethanol (10 mL) was added dropwise within 20 min.
The reaction was refluxed for 3 h. After cooling, the
crude precipitate was filtered off, dried, and recrystallized
from the proper solvent to give compounds 6a–d
respectively.
2-(4-Chlorophenyl)-3-(4-(3-(3,4-dimethoxyphenyl)acryloyl)
phenyl)quinazolin-4(3H)-one (6a).
Brown crystals; yield
67%; m.p. 173–175°C; IR (KBr, cmÀ1): 1708 (C=O,
69.35; H, 4.32; N, 9.49.
unsaturated ketone), 1678 (C=O, cyclic amide), 1655
2-Amino-6-(4-(2-(4-chlorophenyl)-4-oxoquinazolin-3(4H)-
yl)phenyl)-4-(2,5-dimethoxyphen-yl)-4H-pyran-3-carbonitrile
(5b). Yellow crystals; yield 73%; m.p. 122–124°C; IR
1
(C=N), 1638 (C=C), 1255 (OMe), 746 (C-Cl). H-NMR
(DMSO-d6, δ/ppm): 3.80 (s, 3H, OCH3), 4.15 (s, 3H,
OCH3), 6.81–7.80 (m, 17H, Ar-H). 13C-NMR (75 MHz,
DMSO-d6) δ/ppm: 57.2, 57.4 (2OCH3), 110.8, 111.7,
112.4, 113.2, 114.5, 118.6, 119.5, 120.1, 121.7, 125.1,
126.7, 131, 2, 134.7, 135.8, 137.1, 138.9, 140.3, 142.3,
142.7, 147.4, 149.1, 154.2, 156.9 (C=C and C=N), 160.7,
189.5 (2C=O). MS (m/z, (relative abundance, %)): 524
(11), 522 (M+, 33), 421 (59), 210 (74), 133 (100), 91
(KBr, cmÀ1): 3500 (NH2), 2219 (CN), 1683 (C=O, cyclic
1
amide), 1655 (C=N), 1281 (OMe), 744 (C-Cl). H-NMR
(DMSO-d6, δ/ppm): 3.80 (s, 3H, OCH3), 3.94 (s, 3H,
OCH3), 4.20 (d, 1H, CH-Ar, J = 7.3 Hz), 4.95 (d, 1H, CH,
pyran, J = 7.3 Hz), 6.81 (s, 2H, NH2, exchangeable),
6.51–7.80 (m, 15H, Ar-H). MS (m/z, (relative abundance,
%)): 591 (7), 589 (M+, 23), 523 (82), 422 (63), 133 (100),
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

R. S. Gouhar, D. S. A. Haneen, and S. M. El-Hallouty
Vol 000
(86), 77 (56). Anal. Calcd for C31H23ClN2O4: C, 71.19; H,
122.5, 123.9, 124.9, 125.3, 126.2, 129.6, 131.3, 133.1,
135.4, 136.5, 138.9, 141.3, 143.4, 143.6, 144.7, 146.1,
147.4, 149.6, 154.2, 156.7 (C=C and C=N), 160.8 (C=O).
MS (m/z, (relative abundance, %)): 539 (4), 537 (M+, 13),
436 (62), 334 (34), 210 (46), 151 (56), 133 (100), 91 (53),
77 (76). Anal. Calcd for C31H25ClN4O3: C, 69.33; H,
4.69; N, 10.43; Found: C, 69.35; H, 4.70; N, 10.40.
4.43; N, 5.36; Found: C, 71.21; H, 4.40; N, 5.40.
2-(4-Chlorophenyl)-3-(4-(3-(2,5-dimethoxyphenyl)acryloyl)
phenyl)quinazolin-4(3H)-one (6b). Yellow crystals; yield
57%; m.p. 136–138°C; IR (KBr, cmÀ1): 1706 (C=O,
unsaturated ketone), 1675 (C=O, cyclic amide), 1655
1
(C=N), 1625 (C=C), 1255 (OMe), 746 (C-Cl). H-NMR
(DMSO-d6, δ/ppm): 3.82 (s, 3H, OCH3), 4.09 (s, 3H,
OCH3), 6.81–7.80 (m, 17H, Ar-H). MS (m/z, (relative
abundance, %)): 524 (7), 522 (M+, 25), 174 (54), 151
(78), 91 (45), 77 (69), 55 (91), 43 (100). Anal. Calcd for
C31H23ClN2O4: C, 71.19; H, 4.43; N, 5.36; Found: C,
2-(4-Chlorophenyl)-3-(4-(5-(2,5-dimethoxyphenyl)-4,5-
dihydro-1H-pyrazol-3-yl)phenyl)quinazolin-4(3H)-one (7b).
Dark red crystals; yield 59%; m.p. 169–171°C; IR (KBr,
cmÀ1): 3400 (NH), 1683 (C=O, cyclic amide), 1657
1
(C=N), 1235 (OCH3), 749 (C-Cl). H-NMR (DMSO-d6,
71.15; H, 4.47; N, 5.38.
3-(4-(3-(1H-Indol-3-yl)acryloyl)phenyl)-2-(4-chlorophenyl)
δ/ppm): 3.29–3.57 (m, 2H, CH2), 3.50 (s, 3H, OCH3),
3.80 (s, 3H, OCH3), 3.95–4.18 (m, 1H, CH), 7.10–7.81
(m, 15H, Ar-H), 9.82 (br.s, 1H, NH, exchangeable). MS
(m/z, (relative abundance, %)): 539 (11), 537 (M+, 32),
523 (25), 368 (86), 313 (87), 236 (100), 151 (74), 77
(34). Anal. Calcd for C31H25ClN4O3: C, 69.33; H, 4.69;
N, 10.43; Found: C, 69.29; H, 4.73; N, 10.48.
General procedures of synthesis of compounds 7c,d. The
foregoing method was applied using the chalcones 6c or 6d
in the presence of glacial acetic acid (10 mL) upon reflux
for 7 h to give compounds 7c,d, respectively.
quinazolin-4(3H)-one (6c).
Pale brown crystals; yield
76%; m.p. 175–177°C; IR (KBr, cmÀ1): 3448 (NH), 1713
(C=O, unsaturated ketone), 1668 (C=O, cyclic amide),
1
1655 (C=N), 1625 (C=C), 750 (C-Cl). H-NMR (DMSO-
d6, δ/ppm): 7.30–7.61 (m, 19H, Ar-H), 11.80 (br.s, 1H,
NH, exchangeable). 13C-NMR (75 MHz, DMSO-d6)
δ/ppm: 113.5, 114.5, 118.2, 119.5, 121.7, 123.0, 124.9,
125.1, 126.7, 127.3, 128.5, 129.1, 130.5, 131,2, 134.7,
135.8, 137.1, 138.9, 140.3, 142.3, 142.7, 147.4, 149.1,
154.2, 156.9 (C=C and C=N), 160.7, 189.5 (2C=O). MS
(m/z, (relative abundance, %)): 503 (14), 501 (M+, 41),
404 (23), 297 (34), 213 (25), 151 (34), 91 (35), 44 (76),
3-(4-(1-Acetyl-5-(1H-indol-3-yl)-4,5-dihydro-1H-pyrazol-3-
yl)phenyl)-2-(4-chlorophenyl)quinazolin-4(3H)-one (7c).
Yellow crystals; yield 55%; m.p. 165–167°C; IR (KBr,
cmÀ1): 3452 (NH), 1716 (C=O, ketone), 1688 (C=O,
cyclic amide), 1652 (C=N), 751 (C-Cl). 1H-NMR
(DMSO-d6, δ/ppm): 2.16 (s, 3H, COCH3), 3.61–3.90 (m,
2H, CH2), 4.80–5.10 (m, 1H, CH), 7.10–7.81 (m, 17H,
Ar-H), 10.56 (br.s, 1H, NH, exchangeable). 13C-NMR
(75 MHz, DMSO-d6) δ/ppm: 23.7 (CH3-CO), 42.5
(CH2), 51.6 (CH), 119.51, 120.3, 120.7, 121.7, 124.9,
125.0, 126.2, 127.4, 128.2, 129.6, 131.3, 133.7, 135.8,
136.5, 138.9, 141.3, 142.4, 143.6, 144.7, 146.1, 147.4,
149.1, 154.2, 156.9 (C=C and C=N), 160.1, 168.7
(2C=O). MS (m/z, (relative abundance, %)): 560 (20),
558 (M+, 65), 150 (58), 118 (43), 95 (52), 81 (100), 77
(42). Anal. Calcd for C33H24ClN5O2: C, 71.03; H, 4.34;
N, 12.55; Found: C, 71.05; H, 4.36; N, 12.51.
40 (100). Anal. Calcd for C31H20ClN3O2: C, 74.18; H,
4.02; N, 8.37; Found: C, 74.22; H, 4.05; N, 8.35.
2-(4-Chlorophenyl)-3-(4-(3-(naphthalen-2-yl)acryloyl)
phenyl)quinazolin-4(3H)-one (6d).
Brownish yellow
crystals; yield 69%; m.p. 203–205°C; IR (KBr, cmÀ1):
1706 (C=O, unsaturated ketone), 1663 (C=O, cyclic
1
amide), 1655 (C=N), 1625 (C=C), 755 (C-Cl); H-NMR
(DMSO-d6, δ/ppm): 7.30–8.11 (m, 21H, Ar-H); MS (m/z,
(relative abundance, %)): 514 (14), 512 (M+, 35), 379
(45), 174 (53), 173 (67), 151 (100), 137 (36), 77 (15).
Anal. Calcd for C33H21ClN2O2: C, 77.26; H, 4.13; N,
5.46; Found: C, 77.30; H, 4.15; N, 5.50.
General procedures of synthesis of compounds 7a,b.
A
mixture of the chalcone 6a or 6b (0.005 mol) and
hydrazine hydrate 98% (2.5 mL; 0.005 mol) in 25 mL
absolute ethanol was refluxed for 10 h. After cooling, the
crude precipitate was filtered off, dried, and recrystallized
from the proper solvent to give compounds 7a,b,
3-(4-(1-Acetyl-5-(naphthalen-2-yl)-4,5-dihydro-1H-pyrazol-
3-yl)phenyl)-2-(4-chlorophenyl)quinazolin-4(3H)-one (7d).
Brown crystals; yield 73%; m.p. 195–197°C; IR (KBr,
cmÀ1): 3452 (NH), 1714 (C=O, ketone), 1687 (C=O,
cyclic amide), 1656 (C=N), 746 (C-Cl). 1H-NMR
(DMSO-d6, δ/ppm): 1.99 (s, 3H, COCH3), 3.65–3.92 (m,
2H, CH2), 4.90–5.13 (m, 1H, CH), 6.97–7.98 (m, 19H,
Ar-H). MS (m/z, (relative abundance, %)): 571 (12), 569
(M+, 35), 468 (49), 371 (32), 211 (42), 133 (100), 91
(35), 77 (65). Anal. Calcd for C35H25ClN4O2: C, 73.87;
H, 4.43; N, 9.85; Found: C, 73.88; H, 4.45; N, 9.83.
respectively.
2-(4-Chlorophenyl)-3-(4-(5-(3,4-dimethoxyphenyl)-4,5-
dihydro-1H-pyrazol-3-yl)phenyl) quinazolin-4(3H)-one (7a).
Reddish brown crystals; yield 75%; m.p. 211–213°C; IR
(KBr, cmÀ1): 3400 (NH), 1680 (C=O, cyclic amide),
1651 (C=N), 1235 (OCH3), 749 (C-Cl). 1H-NMR
(DMSO-d6, δ/ppm): 3.31–3.60 (m, 2H, CH2), 3.70 (s, 3H,
OCH3), 3.81 (s, 3H, OCH3), 4.10–4.13 (m, 1H, CH),
7.10–7.81 (m, 15H, Ar-H), 9.80 (br.s, 1H, NH,
exchangeable). 13C-NMR (75 MHz, DMSO-d6) δ/ppm:
42.5 (CH2), 51.6 (CH), 56.1, 56.5 (2OCH3), 119.1, 120.9,
General procedures of synthesis of compounds 8a,b.
A
mixture of the chalcone 6a or 6b (0.005 mol) and urea
(0.5 g; 0.005 mol) in 20 mL ethanol and concentrated
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2019
Synthesis and Anticancer Evaluation of Some Novel Quinazolin-4(3H)-one
Derivatives
HCl was refluxed for 7 h. The reaction mixture was
concentrated to half of its volume under reduced
pressure. After cooling, it was neutralized with
ammonium hydroxide solution at 10°C. The crude
precipitate was filtered off, washed with water, dried, and
recrystallized from the proper solvent to give compounds
8a,b, respectively.
137 (34), 91 (36), 77 (52). Anal. Calcd for
C32H20ClN5OS: C, 68.87; H, 3.61; N, 12.55; Found: C,
68.90; H, 3.59; N, 12.57.
2-(4-Chlorophenyl)-3-(4-(6-(naphthalen-2-yl)-2-thioxo-1,2-
dihydropyrimidin-4-yl)phenyl)quinazolin-4(3H)-one (8d).
Brownish yellow crystals; yield 57%; m.p. 148–150°C;
IR (KBr, cmÀ1): 3400, 3323 (NH), 1702 (C=O, cyclic
1
amide), 1647 (C=N), 1220 (C=S), 750 (C-Cl). H-NMR
2-(4-Chlorophenyl)-3-(4-(6-(3,4-dimethoxyphenyl)-2-oxo-
(500 MHz, DMSO-d6, δ/ppm): 7.10–7.81 (m, 20H, Ar-H),
13.09 (br.s, 1H, NH, exchangeable). 13C-NMR (75 MHz,
DMSO-d6) δ/ppm: 122.1, 122.7, 123.5, 124.7, 126.1,
126.9, 127.3, 128.3, 129.6, 131.9, 132.6, 133.8, 135.1,
136.2, 137.5, 138.2, 139.6, 140.6, 141.6, 142.8, 143.2,
144.7, 145.6, 146.1, 146.4, 148.7, 149.9, 155.5 (C=C and
C=N), 160.4 (C=O), 180.7 (C=S). MS (m/z, (relative
abundance, %)): 571 (20), 569 (M+, 63), 568 (47), 444
(32), 313 (45), 210 (52), 133 (100), 91 (65), 77 (75).
Anal. Calcd for C34H21ClN4OS: C, 71.76; H, 3.72; N,
9.85; Found: C, 71.75; H, 3.70; N, 9.90.
1,2-dihydropyrimidin-4-yl)
phenyl)quinazolin-4(3H)-one
(8a). Brown crystals; yield 57%; m.p. 187–189°C; IR
(KBr, cmÀ1): 3443(NH), 1701, 1684 (2C=O, cyclic
1
amide), 1655 (C=N), 1281 (OCH3), 753 (C-Cl). H-NMR
(DMSO-d6, δ/ppm): 3.73 (s, 3H, OCH3), 3.79 (s, 3H,
OCH3), 5.86 (s, 1H, CH-pyrimidine), 7.10–7.81 (m, 15H,
Ar-H), 11.07 (br.s, 1H, NH, exchangeable). 13C-NMR
(75 MHz, DMSO-d6) δ/ppm: 56.3, 56.5(2OCH3), 121.8,
122.2, 123.5, 124.7, 125.1, 126.9, 128.3, 129.6, 130.9,
131.7, 133.1, 135.4, 136.5, 138.2, 141.0, 143.5, 143.8,
144.7, 145.6, 146.1, 147.4, 148.2, 149.6, 156.7 (C=C and
C=N), 160.1, 160.6 (2C=O). MS (m/z, (relative
abundance, %)): 565 (15), 563 (M+, 45), 261 (43), 233
(100), 218 (34), 172 (42), 158 (25), 144 (32), 91 (23), 77
(52). Anal. Calcd for C32H23ClN4O4: C, 68.27; H, 4.12;
N, 9.95; Found: C, 68.30; H, 4.15; N, 9.91.
BIOLOGICAL ASSAY
Cell culture. MCF-7 cell lines were obtained from the
Karolinska Institute, Stockholm, Sweden. HEPG2 cells
were maintained in RPMI 1640 medium, while MCF-7 Hu-
man cancer cells were maintained in DMEM medium
(Lonza Biowahittkar, Belgium). All the media were supple-
mented with 1% antibiotic–antimycotic mixture [10,000 U/
mL potassium penicillin, 10,000 μg/mL streptomycin sul-
fate, 25 μg/mL amphotericin B, and 1% L-glutamine
(Biowest, USA)].
2-(4-Chlorophenyl)-3-(4-(6-(2,5-dimethoxyphenyl)-2-oxo-
1,2-dihydropyrimidin-4-yl)
(8b).
phenyl)quinazolin-4(3H)-one
Brownish yellow crystals; yield 62%; m.p. 155–
157°C; IR (KBr, cmÀ1): 3406 (NH), 1699, 1681 (2C=O,
cyclic amide), 1658 (C=N), 1281 (OCH3), 753 (C-Cl).
1H-NMR (DMSO-d6, δ/ppm): 3.49 (s, 3H, CH3, OCH3),
3.70 (s, 3H, OCH3), 5.99 (s, 1H, CH-pyrimidine), 7.10–
7.81 (m, 15H, Ar-H), 11.03 (br.s, 1H, NH, exchangeable).
MS (m/z, (relative abundance, %)): 565 (6), 563 (M+, 19),
542 (61), 441 (32), 313 (45), 212 (31), 133 (100), 91 (54),
77 (67). Anal. Calcd for C32H23ClN4O4: C, 68.27; H,
4.12; N, 9.95; Found: C, 68.24; H, 4.13; N, 9.89.
MTT cytotoxicity assay. Cell viability was investigated
using
MTT
[3-(4,5-dimethylthiazol-2-yl)-2,
5-
diphenyltetra-zolium bromide] (Bio Basic Canada Inc.,
Canada). This reaction depends on mitochondrial reduction
of yellow MTT into purple formazan. All the preceding
steps were carried out in sterile laminar airflow cabinet Bio-
safety class II level (Baker, SG403INT; Sanford, ME,
USA). All incubations were performed at 37°C in 5% CO2
incubator in the humidified atmosphere (Sheldon,
TC2323; Cornelius, OR, USA). Cells were seeded into 96-
well microtiter plastic plates at the concentration of (104
cells per well) and allowed to adhere for 24 h. Medium
was aspirated and fresh medium (without serum) was added
to the cells with 100 μg/mL in dimethyl sulfoxide, and then,
the promising compounds which give 100% inhibition will
be subjected to various concentrations of the promising
compounds (100, 50, 25, 12.5, 6.25, 3.12, 1.56, and
0.78 μg/mL in DMSO) and incubated for 48 h. The medium
was aspirated and 40 μL MTT salts (2.5 μg/mL) was added
to each correctly and incubated for a further 4 h. To stop the
reaction and dissolve any formed formazan crystals, 200 μL
of 10% sodium dodecyl sulfate were added to each well and
General procedures of synthesis of compounds 8c,d. The
foregoing method was applied using the chalcones 6c or 6d
and thiourea (0.5 g) in the presence of NaOH (0.5 g) in
5 mL of water instead of urea. The mixture was refluxed
for 6 h in ethanol (25 mL), concentrated, and neutralized
with dilute HCl and the formed precipitate was filtered,
washed with water, dried, and crystallized to give
compounds 8c,d, respectively.
3-(4-(6-(1H-Indol-3-yl)-2-thioxo-1,2-dihydropyrimidin-4-yl)
phenyl)-2-(4-chlorophenyl)quinazolin-4(3H)-one
(8c).
Reddish brown crystals; yield 63%; m.p. 135–137°C; IR
(KBr, cmÀ1): 3444, 3328 (NH), 1691 (C=O, cyclic
1
amide), 1651 (C=N), 1150 (C=S), 752 (C-Cl). H-NMR
(DMSO-d6, δ/ppm): 7.10–7.81 (m, 16H, Ar-H), 11.93 (br.
s, 1H, NH-indolyl, exchangeable), 13.11 (br.s, 1H, NH-
C=S, exchangeable). MS (m/z, (relative abundance, %)):
560 (11), 558 (M+, 33), 368 (34), 328 (78), 313 (24), 286
(34), 262 (52), 191 (76), 180 (100), 163 (63), 151 (52),
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

R. S. Gouhar, D. S. A. Haneen, and S. M. El-Hallouty
Vol 000
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incubated overnight at 37°C. The amount of formazan prod-
uct was measured at 595 nm with a reference wavelength of
620 nm as a background using a microplate reader (Bio-Rad
Laboratories, model 3350, USA). For the untreated cells
(negative control), medium was added instead of the test
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CONCLUSION
Some quinazolinone derivatives were synthesized for
the purpose of anticancer cell line evaluation. The
quinazolinones which bearing indolyl and/or pyridinyl
moieties in their side chain showed highest potency
cytotoxic activity against the MCF-7 and HepG2 cancer
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Acknowledgment. The authors thank the National Research
Centre for the financial support (Project number 11010312).
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Journal of Heterocyclic Chemistry
DOI 10.1002/jhet