Synthesis and Reactions of Some New Quinazoline Derivatives for In Vitro Evaluation as Anticancer and Antimicrobial Agents

Article abstract of DOI:10.1002/jhet.3248

A novel series of N,O,S heterocyclic compounds incorporated at position-3 of a quinazolin-4(3H)-one moiety were synthesized through a p-phenloxy binding including oxadiazolethione pyridazinetrione, thiazolidinedione, benzothiazindione, and quinazolinedione. New carbamates, urea derivatives, and Mannich bases were also prepared for the purpose of in vitro anticancer cell lines evaluation and cytotoxic activity. The key intermediate 2-(4-chlorophenyl)-3-(4-hydroxyphenyl)quinazolin-4(3H)-one was prepared from the reaction of 4H-benzo [d][3,1]oxazin-4-one and a nitrogenous nucleophilic compound, 4-aminophenol. Some of the newly synthesized compounds showed significant cytotoxic activity.

Full text of DOI:10.1002/jhet.3248

Month 2018 Synthesis and Reactions of Some New Quinazoline Derivatives for In Vitro
Evaluation as Anticancer and Antimicrobial Agents
*
Rasha S. Gouhar
and Mohsen M. Kamel
Therapeutical Chemistry Department, National Research Centre, Dokki, P.O. 12622, Giza, Egypt
*
E-mail: dr.rasha_gouhar@yahoo.com
Received March 7, 2018
DOI 10.1002/jhet.3248
Published online 00 Month 2018 in Wiley Online Library (wileyonlinelibrary.com).
A novel series of N,O,S heterocyclic compounds incorporated at position-3 of a quinazolin-4(3H)-one
moiety were synthesized through a p-phenloxy binding including oxadiazolethione pyridazinetrione,
thiazolidinedione, benzothiazindione, and quinazolinedione. New carbamates, urea derivatives, and
Mannich bases were also prepared for the purpose of in vitro anticancer cell lines evaluation and cytotoxic
activity. The key intermediate 2-(4-chlorophenyl)-3-(4-hydroxyphenyl)quinazolin-4(3H)-one was prepared
from the reaction of 4H-benzo [d][3,1]oxazin-4-one and a nitrogenous nucleophilic compound, 4-
aminophenol. Some of the newly synthesized compounds showed significant cytotoxic activity.
J. Heterocyclic Chem., 00, 00 (2018).
INTRODUCTION
The aim of the present investigation is to synthesize a
new series of N-3-substituted-quinazoline-4(3H)-one
derivatives incorporated to N,O,S heterocyclic derivatives
at position-3 of the 2-(p-chlorophenyl)-quinazolin-4(3H)-
one backbone. The antimicrobial activity of the prepared
derivatives was evaluated.
Quinazolines and quinazolone derivatives were reported
to exhibit antimalarial [1], antimicrobial [2], anti-
inflammatory [3], anticonvulsant [4], antihypertensive [5],
antidiabetic [6], and anticancer activities [7–9]. Certain
quinazoline derivatives were found to inhibit various
forms of cellular phosphorylation [10]. Some derivatives
demonstrated significant central nervous system activity
through acting as ligands for benzodiazepine and amino
butyric acid receptors [11]. They also acted as DNA
binding agents as well as effective adrenergic blockers
[12]. On the other hand, several quinazolinone
derivatives are reported to constitute the building blocks
for many natural alkaloids isolated from plant kingdom
or from microorganisms such as febrifugine (I) (Fig. 1), a
quinazoline alkaloid that possesses antimalarial potential
from the Chinese plant Dichroa febrifuga [13]. Recently,
in our drug discovery program, we synthesized [14,15]
a new class of 10H-spiro[cyclohexane-1,20-quinazolin]-
40(30H)-one analogs (II and III) as PARP-1 inhibitors
(Fig. 1). The derivatives that exhibited the best-fit
target enzyme according to the docking study were
selected to assess their in vitro cytotoxic potency and
antiproliferative activity against breast cancer cell line;
MCF7. In addition, they showed effective inhibitory
effect on the target PARP-1 enzyme.
RESULTS AND DISCUSSION
Chemistry.
Reaction of anthranilic acid in pyridine
with p-chlorobenzoyl chloride at room temperature
afforded the starting material 2-(4-chlorophenyl)-
4H-benzo[d][3,1]oxazin-4-one (1). The reaction of
benzoxazine derivative 1 with a nitrogenous nucleophilic
compound, 4-aminophenol, in glacial acetic acid and
anhydrous sodium acetate under reflux for 15 h, yielded
the key intermediate material quinazolinone derivative,
namely, 2-(4-chlorophenyl)-3-(4-hydroxyphenyl)quinazolin-
4(3H)-one (2) in a good yield (75%).
Compound
2
reacted with p-formaldehyde and
appropriate secondary amines, namely, diethylamine,
morpholine, and methyl piperazine in ethanolic solution
to give the corresponding Mannich bases 3a–c,
respectively. The presence of the Mannich side chain at
the ortho position of the phenolic hydroxyl group may
overcome the insolubility problem of such compounds
© 2018 Wiley Periodicals, Inc.

R. S. Gouhar and M. M. Kamel
Vol 000
Figure 1. The structures of quinazolinone analogs I, II, and III.
through the formation of hydrochloride salt (Scheme 1).
Furthermore, the reaction of 2 with ethyl bromoacetate in
the presence of anhydrous potassium carbonate and dry
acetone for 12 h provided the corresponding ethyl 2-(4-
(2-(4-chlorophenyl)-4-oxoquinazolin-3(4H)-yl)phenoxy)
acetate (4), which upon reaction with hydrazine hydrate
in ethanol for 3 h, afforded the corresponding 2-(4-(2-
(4-chlorophenyl)-4-oxoquinazolin-3(4H)-yl)phenoxy)
acetohydrazide (5) (Scheme 1). Cyclocondensation of the
intermediate acetohydrazide 5 with either carbon disulfide
in the presence of KOH or with diethyl oxalate yielded the
The reaction of 5 with sodium nitrite in acetic acid at
2–5°C in ice bath afforded the corresponding carbonyl
azide 8, which was further reacted with thioglycolic
acid in dry benzene to provide the corresponding
thiazolidinedione derivative 9. This intermediate
8
reacted with thiosalicylic acid in dry benzene providing
the corresponding benzothiazinedione derivative 10
(Scheme 2).
The reaction of the carbonyl azide 8 with anthranilic
acid in dry dioxane afforded the diquinazolinone
derivative 11. The carbamate derivatives 12a–c were
obtained upon the reaction of the carbonyl azide 8 with
phenols, namely, phenol, 4-aminophenol, and 4-
bromophenol in dry benzene for 6 h (Scheme 2). Finally,
target 1,3,4-oxadiazol-2(3H)-thione derivative
6
and
pyridazine-3,4,6-trione
(Scheme 1).
analogue
7,
respectively
Scheme 1. Formation of the quinazolinone derivatives 2–7.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2018
Synthesis and Anticancer Evaluation of Some New Quinazoline Derivatives
Scheme 2. Synthesis and reactions of the azide derivative 8. Formation of
the quinazolinone derivatives 9–13.
compound showed varying degrees of inhibition against
the tested microorganisms. Susceptibilities of fungal and
bacterial isolates to different chemical compounds were
investigated by measuring their inhibitory effect on the
growth, compared with the solvent used. There is relation
between chemical structure and activity where
acetohydrazide derivative 5, diazine derivative 7, and the
azide derivative 8 exhibited high antibacterial and
antifungal activities, hydroxyquinazoline drivatives as 2,
3b and the thiazole and thiazine derivatives as 9, 10
exhibited moderate antibacterial and antifungal activities
compared with the standard drugs, while compounds 1, 4,
6, 11, 12c, 13b exhibited no antimicrobial activity (Cf.,
Table 1).
Evaluation of the antitumor activity.
This study
exhibited the antitumor activity of some of the newly
synthesized compounds. Ten derivatives were selected as
representative examples to examine their in vitro
antitumor activities against Hep-G2 human liver cancer
cell line and MCF-7 human breast cancer cell line. The
effective dose calculated as IC50, which correspond to the
compound concentration resulted in 50% mortality in the
total cells count and presented in (Table 2). The activities
of the derivatives against carcinoma cells were compared
with the cytotoxicity of doxorubicin as a reference drug.
The results showed that the benzothiazindione and
oxoquinazoline derivatives 10, 12a, respectively,
exhibited the highest cytotoxic activity against the liver
Table 1
Antibacterial and antifungal activity (as inhibition zone in mm diameter).
No inhibition zone diameter (mm/mg sample)
Candida Aspergillus
the reaction of the carbonyl azide 8 with different aniline
derivatives, namely, aniline, 4-bromoaniline, and 4-
chloroaniline in dry dioxane, provided the corresponding
substituted phenyl urea derivatives 13a–c, respectively
(Scheme 2).
Structures of all new compounds were illustrated by
chemical microanalyses and spectroscopic analyses, IR,
1H NMR, and mass spectra.
Staphylococcus Escherichia albicans
Niger
aureus G+
coli Gꢀ (fungus) (fungus)
1
2
3b
4
5
6
7
8
0
13
16
0
29
0
24
28
16
12
0
0
10
8
0
24
0
22
23
20
8
0
10
11
0
0
0
0
0
14
0
15
0
11
12
13
11
0
10
10
12
14
0
9
10
11
ANTIMICROBIAL EVALUATION
0
12c
13b
Tetracycl.
Am B
DMSO
0
0
28
0
0
30
0
0
0
0
The synthesized compounds were evaluated for their
in vitro antimicrobial activity using gram-positive
bacteria (Staphylococcus aureus), gram-negative bacteria
(Escherichia coli), and two fungus (yeast Candida
albicans and mold Aspergillus niger); Tetracycline,
19
0
16
0
0
0
Inhibition zone diameter (mm). Tetracycl., Tetracycline (antibacterial
agent), AmB, Amphotericin B (antifungal agent). The concentration of all
synthesized compounds and the reference was (5 mg/1 mL of DMSO).
Zone of inhibition: 0–11 mm (low); 11–15 mm (moderate); >15 mm
(high); 0.0 = no inhibition.
Amphotericin
B were used as standard drugs for
antibacterial and antifungal activities, respectively. The
results revealed that majority of the synthesized
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

R. S. Gouhar and M. M. Kamel
Vol 000
Table 2
2-(4-Chlorophenyl)-4H-benzo[d][3,1]oxazin-4-one (1).
Cytotoxic activity of some compounds against human tumor cells.
To a solution of anthranilic acid (1.4 g; 1.1 mmol)
dissolved in pyridine (60 mL), p-chlorobenzoyl chloride
(2 mmol) was added. The mixture was stirred for 40 min
at room temperature. This mixture was poured into a
beaker containing 5% sodium bicarbonate (50 mL)
solution with stirring. The obtained solid was
recrystallized from ethanol. The purity of the prepared
compound was checked by TLC using benzene: EtOAc
(7:3) as the mobile phase. I2 vapor was used as
visualizing agent.
Yellow crystals; yield 80%; mp 152–153°C as reported;
IR (KBr, cmꢀ1): 1756 (C═O), 1637 (C═N); 1H NMR
(DMSO-d6, δ ppm): 7.30–8.13 (m, 8H, Ar─H); MS (m/z,
(relative abundance, %)): 259 (M + 2, 16); 257 (M+, 45);
Anal. Calcd for C14H8ClNO2: C, 65.26; H, 3.13; N, 5.44;
Found: C, 65.30; H, 3.18; N, 5.40.
In vitro cytotoxicity IC50 (μM)•
No.
Compounds
HePG2
MCF-7
DOX
1
2
3a
4
7
8
9
10
11
12a
4.50 0.2
59.2 3.34
89.1 4.2
48.13 2.89
67.1 3.7
18.18 1.3
12.22 1.1
58.63 3.4
8.1 0.7
4.17 0.2
9.77 0.9
1
2
3
4
5
6
7
8
9
66.31 5.31
8.67 0.63
73.15 4.5
12.82 2.2
8.93 1.96
80.58 4.4
55.66 3.4
85.13 4.21
67.18 3.8
91.45 5.3
9.35 1.0
10
•IC50 (μM):1–10 (very strong), 11–20 (strong), 21–50 (moderate),
51–100 (weak), and above 100 (noncytotoxic).
cancer cells (HepG-2) while they showed moderate
potency against the breast carcinoma cell lines MCF-7
were hydroxy quinazoline 2, 3a. Fortunately, dual
inhibition potency against both HepG-2 and MCF-7
cancer cell lines were gained by the diazintrione and
azide derivatives in compounds 7, 8. The rest of the
tested derivatives produced weak activity as 1, 4, 9, 11.
2-(4-Chlorophenyl)-3-(4-hydroxyphenyl)quinazolin-4(3H)-
one (2). A mixture of compound 1 (2.58 g, 0.01 mol) and
p-aminophenol (1.10 g, 0.01 mol) in 20 mL glacial acetic
acid in presence of anhydrous sodium acetate was
refluxed for 15 h, after cooling, the crude mass was
crystallized from ethanol to give compound 2.
Violet crystals; yield 83%; mp 179–180°C; IR (KBr,
1
cmꢀ1): 3430 (OH), 1712 (C═O), 1680 (C═N); H NMR
(DMSO-d6, δ ppm): 7.90–8.33 (m, 12H, Ar─H), 9.20
(br, s, 1H, OH, exchangeable with D2O); MS (m/z,
(relative abundance, %)): 350 (M + 2, 11); 348 (M+, 35);
Anal. Calcd for C20H13ClN2O2: C, 68.87; H, 3.76; N,
8.03; Found: C, 68.54; H, 3.98; N, 8.14.
EXPERIMENTAL
All melting points were used uncorrected and were taken
with open capillary tubes using electrothermal apparatus
9100. Elemental microanalyses were carried out at
microanalytical unit, Central Services Laboratory,
National Research Centre, Dokki, Giza-Egypt, using Vario
Elementar and were found within + or ꢀ0.5% of the theo-
retical values. Infrared spectra were recorded on a Jasco
FT/IR-6100, Fourier Transform Infrared Spectrometer at
cmꢀ1 scale using KBr disc technique at the Central
Services Lab. NRC, Dokki, Giza, Egypt. 1H NMR spectra
were determined by using a JEOL EX-NMR Spectrometer-
300 MHz- at Central Services Lab, (Faculty of Science,
Cairo University, Cairo, Egypt). Mass spectra were mea-
sured with Finnigan MAT SSQ-7000 mass spectrometer
at the Central Services, NRC Dokki, Giza, Egypt.
Follow-up of the reactions and checking the purity of the
prepared compounds were made by thin-layer chromatog-
raphy (TLC) on silica gel-precoated aluminum sheets
(Type 60 F254-Merck, Darmstadt, Germany), and the
spots were detected by exposure to UV Lamp at 254 nm.
The IUPAC system was followed in the nomenclature of
the new compounds. The antimicrobial activity was carried
out in Micro Analytical Center at Cairo University. The an-
ticancer activity was carried out in pharmacological unit of
national cancer institute for performing the anticancer
screening.
General procedures for the synthesis of Mannich bases
3a–c. A mixture of paraformaldehyde (0.09 g; 1 mmol),
the appropriate amine, namely, diethylamine, morpholine,
or methylpiperazine (15 mmol) and two drops of pyridine
in 20 mL ethanol was refluxed for 30 min till solubility
of p-formaldehyde. Then, a solution of 2 (0.35 g;
0.1 mmol) in ethanol (2 mL) was added to the mixture
and refluxed for 8 h. After cooling, the formed precipitate
was filtered off, dried, and recrystallized from dioxane to
give Mannich bases 3a–c.
2-(4-Chlorophenyl)-3-(3-((diethylamino)methyl)-4-
hydroxyphenyl)quinazolin-4(3H)-one (3a). Yellow crystals;
yield 65%; mp 105–106°C; IR (KBr, cmꢀ1): 3427 (OH),
1718 (C═O), 1650 (C═N), 1217 (OEt); 1H NMR
(DMSO-d6, δ ppm): 2.12 (t, 6H, 2CH3), 3.20 (q, 4H,
2CH2), 4.20 (s, 2H, CH2), 6.61–8.30 (m, 11H, Ar─H),
9.13 (br. s, 1H, OH, exchangeable with D2O); MS (m/z,
(relative abundance, %)): 435 (M + 2, 4); 433 (M+, 15);
Anal. Calcd for C25H24ClN3O2: C, 69.20; H, 5.57; N,
9.68; Found: C, 69.60; H, 5.60; N, 9.50.
2-(4-Chlorophenyl)-3-(4-hydroxy-3-(morpholinomethyl)
phenyl)quinazolin-4(3H)-one (3b). Yellow crystals; yield
73%; mp 139–140°C; IR (KBr, cmꢀ1): 3427 (OH), 1728
1
(C═O), 1650 (C═N); H NMR (DMSO-d6, δ ppm): 2.80
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2018
Synthesis and Anticancer Evaluation of Some New Quinazoline Derivatives
(t, 4H, 2CH2─N), 3.50 (t, 4H, 2CH2─O), 3.70 (s, 2H,
CH2), 6.61–7.30 (m, 11H, Ar─H), 9.30 (br. s, 1H, OH,
exchangeable with D2O); MS (m/z, (relative abundance,
%)): 449 (M + 2, 8), 447 (M+, 22); Anal. Calcd for
C25H22ClN3O3: C, 67.04; H, 4.95; N, 9.38; Found: C,
cooling, concentrated HCl neutralized the crude
precipitate. The separated solid was filtered off, washed
with water, dried, and recrystallized from the ethanol to
provide compound 6.
Yellowish brown crystals; yield 70%; mp 144–146°C;
IR (KBr, cmꢀ1): 3320 (NH), 1718 (C═O), 1655 (C═N);
1H NMR (DMSO-d6, δ ppm): 3.30 (s, 2H, CH2), 6.81–
7.80 (m, 12H, Ar─H), 10.50 (br. s, 1H, NH,
exchangeable with D2O); MS (m/z, (relative abundance,
%)): 464 (M + 2, 11), 462 (M+, 35); Anal. Calcd for
C23H15ClN4O3S: C, 59.68; H, 3.27; N, 12.10; Found: C,
59.70; H, 3.30; N, 12.07.
67.10; H, 4.90; N, 9.40.
2-(4-Chlorophenyl)-3-(4-hydroxy-3-((4-methylpiperazin-1-yl)
methyl)phenyl)quinazolin-4(3H)-one (3c). Yellow crystals;
yield 58%; mp 111–112°C; IR (KBr, cmꢀ1): 3413 (OH),
1
1724 (C═O), 1650 (C═N); H NMR (DMSO-d6, δ/ppm):
2.80 (t, 4H, 2CH2), 3.45 (s, 3H, CH3), 3.50 (t, 4H, 2CH2),
3.70 (s, 2H, CH2), 6.61–7.30 (m, 11H, Ar─H), 9.10 (br.
s, 1H, OH, exchangeable with D2O); MS (m/z, (relative
abundance, %)): 462 (M + 2, 10), 460 (M+, 31); Anal.
Calcd for C26H25ClN4O2: C, 67.75; H, 5.47; N, 12.15;
Found: C, 67.80; H, 5.45; N, 12.19.
Synthesis of 5-(4-(2-(4-chlorophenyl)-4-oxoquinazolin-
3(4H)-yl)phenoxy)dihydropyridazine-3,4,6(5H)-trione (7).
A mixture of the hydrazide 5 (0.42 g; 1 mmol), diethyl
oxalate (0.15 mL; 1.1 mmol), and Na metal (0.025 g) in
absolute ethanol was refluxed for 12 h. After cooling,
HCl acidified the reaction mixture and the crude
precipitate was filtered off, dried, and recrystallized from
ethanol to give compound 7.
Synthesis
of
ethyl
2-(4-(2-(4-chlorophenyl)-4-
oxoquinazolin-3(4H)-yl)phenoxy)acetate (4). A mixture of
2 (3.48 g; 10 mmol, ethyl bromoacetate (2.49 mL;
15 mmol)), anhydrous potassium carbonate (2 g;
14 mmol) in 20 mL dry acetone was refluxed for 12 h.
While hot, the reaction mixture was filtered and the
filtrate was concentrated under vacuum to yield the crude
product that was filtered off, washed several times with
petroleum ether, and recrystallized from ethyl acetate to
provide compound 4.
Yellowish brown crystals; yield 82%; mp 181–182°C;
IR (KBr, cmꢀ1): 1735, 1718 (C═O), 1650 (C═N); 1H
NMR (DMSO-d6, δ ppm): 2.80 (t, 3H, CH3), 3.50 (q,
2H, CH2), 4.20 (s, 2H, CH2), 6.91–8.30 (m, 12H,
Ar─H); MS (m/z, (relative abundance, %)): 436 (M + 2,
4), 434 (M+, 12); Anal. Calcd for C24H19ClN2O4: C,
66.29; H, 4.40; N, 6.44; Found: C, 66.31; H, 4.38; N, 6.49.
Yellowish brown crystals; yield 68%; mp 110–112°C;
IR (KBr, cmꢀ1): 3446 (NH), 1728, 1719, 1690 (C═O),
1
1655 (C═N); H NMR (DMSO-d6, δ ppm): 4.50 (s, 1H,
CH), 6.81–7.80 (m, 12H, Ar─H), 8.50 (br. s, 1H, NH,
exchangeable with D2O), 9.20 (br. s, 1H, NH,
exchangeable with D2O); MS (m/z, (relative abundance,
%)): 476 (M + 2, 10), 474 (M+, 29); Anal. Calcd for
C24H15ClN4O5: C, 60.70; H, 3.18; N, 11.80; Found: C,
60.74; H, 3.20; N, 11.77.
Synthesis of 2-(4-(2-(4-chlorophenyl)-4-oxoquinazolin-
3(4H)-yl)phenoxy)acetyl azide (8).
A stirred carboxylic
acid hydrazide 5 (4.2 g; 10 mmol) in HCl (10%) (20 mL)
at 0–5°C sodium nitrite (10%) was added portion wise
till effervesces ceased. The reaction mixture was stirred
for 1 h. The resulting solid was collected, filtered,
washed with water, dried, and recrystallized from ethanol
to yield compound 8.
Synthesis of 2-(4-(2-(4-chlorophenyl)-4-oxoquinazolin-
3(4H)-yl)phenoxy)acetohydrazide (5).
A mixture of
hydrazine hydrate 99% (1.6 mL; 50 mmol) and a solution
of 4 (4.18 g; 10 mmol) in absolute ethanol (20 mL) was
refluxed for 3 h. The mixture was concentrated, cooled,
and the formed precipitate was filtered off, dried, and
recrystallized from ethanol to provide compound 5.
Yellow crystals; yield 73%; mp 131–133°C; IR (KBr,
1
cmꢀ1): 2307 (N3), 1712, 1677 (C═O), 1655 (C═N); H
NMR (DMSO-d6, δ ppm): 450 (s, 2H, CH2), 6.51–7.20
(m, 12H, Ar─H); MS (m/z, (relative abundance, %)): 433
Yellowish brown crystals; yield 66%; mp 130–131°C;
(M
+
2, 4), 431 (M+, 12); Anal. Calcd for
IR (KBr, cmꢀ1): 3451, 3330, 3215 (NH, NH2), 1722,
1
C22H14ClN5O3: C, 61.19; H, 3.27; N, 16.22; Found: C,
61.20; H, 3.30; N, 16.25.
1667 (C═O), 1655 (C═N); H NMR (DMSO-d6, δ ppm):
3.50 (s, 2H, CH2), 5.31 (br. s, 2H, NH2, exchangeable
with D2O), 6.51–7.20 (m, 12H, Ar─H), 8.51 (br. s, 1H,
NH, exchangeable with D2O); MS (m/z, (relative
abundance, %)): 422 (M + 2, 8), 420 (M+, 25); Anal.
Calcd for C22H17ClN4O3: C, 62.79; H, 4.07; N, 13.31;
Found: C, 62.81; H, 4.10; N, 13.29.
3-((4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)
phenoxy)methyl)thiazolidine-2,4-dione (9).
A mixture of
the azide derivative 8 (0.43 g; 1 mmol) and thioglycolic
acid (0.12 g, 2 mmol) in 15 mL of dry benzene was
refluxed for 1 h through which the reaction mixture was
checked with TLC till the full consumption of 8. The
mixture was concentrated under vacuum, cooled, filtered,
and crystallized from benzene to provide compound 9.
Yellow crystals; yield 79%; mp 188–189°C; IR (KBr,
Synthesis
of
2-(4-chlorophenyl)-3-(4-((5-thioxo-4,5-
dihydro-1,3,4-oxadiazol-2-yl)methoxy)phenyl)quinazolin-
4(3H)-one (6). A mixture of 5 (0.42 g; 1 mmol), KOH
(0.056 g; 1 mmol), and CS2 (1 mL) in 30-mL absolute
ethanol was refluxed for 12 h. After concentration and
1
cmꢀ1): 1780, 1735, 1723 (C═O), 1655 (C═N); H NMR
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

R. S. Gouhar and M. M. Kamel
Vol 000
(DMSO-d6, δ ppm): 3.60 (s, 2H, CH2), 4.60 (s, 2H, CH2),
6.51–7.20 (m, 12H, Ar─H); MS (m/z, (relative abundance,
%)): 479 (M + 2, 13), 477 (M+, 42); Anal. Calcd for
C24H16ClN3O4S: C, 60.31; H, 3.37; N, 8.79; Found: C,
2H, NH2, exchangeable with D2O), 7.10–7.81 (m,
16H, Ar─H), 9.83 (br. s, 1H, NH, exchangeable with
D2O); MS (m/z, (relative abundance, %)): 514 (M + 2,
9), 512 (M+, 25); Anal. Calcd for C28H21ClN4O4: C,
65.56; H, 4.13; N, 10.92; Found: C, 65.52; H, 4.15;
60.35; H, 3.40; N, 8.80.
3-((4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)
phenoxy)methyl)-2H-benzo[e][1,3]-thiazine-2,4(3H)-dione
N, 10.90.
4-Bromophenyl((4-(2-(4-chlorophenyl)-4-oxoquinazolin-
3(4H)-yl)phenoxy)methyl)carbamate (12c). Yellow crystals;
(10).
The foregoing procedures for the preparation of
yield 62%; mp 129–130°C; IR (KBr, cmꢀ1): 3404
(NH), 1716, 1689 (C═O), 1655 (C═N), 630 (C─Br);
1H NMR (DMSO-d6, δ ppm): 3.90 (s, 2H, CH2),
7.10–7.71 (m, 16H, Ar─H), 10.60 (br. s, 1H, NH,
exchangeable with D2O); MS (m/z, (relative abundance,
%)): 577 (M + 2, 5), 575 (M+, 13); Anal. Calcd for
C28H19BrClN3O4: C, 58.30; H, 3.32; N, 7.28; Found:
C, 58.26; H, 3.33; N, 7.33.
compound 9 were applied except that 1.5 mmol of
thiosalicylic acid was used instead of thioglycolic acid to
give compound 10.
Yellow crystals; yield 63%; mp 122–124°C; IR (KBr,
1
cmꢀ1): 1755,1733, 1718 (C═O), 1655 (C═N); H NMR
(DMSO-d6, δ ppm): 4.20 (s, 2H, CH2), 6.51–7.20 (m,
16H, Ar─H); MS (m/z, (relative abundance, %)): 541
(M
+
2, 12), 539 (M+, 35); Anal. Calcd for
C29H18ClN3O4S: C, 64.50; H, 3.36; N, 7.78; Found: C,
64.48; H, 3.35; N, 7.80.
General procedures for the synthesis of 13a–c.
A
mixture of the azide derivative 8 (2.1 g; 5 mmol) and the
appropriate anilines, namely, aniline, 4-bromoaniline, or
4-chloroaniline (5 mmol), in dry dioxane (20 mL) was
refluxed for 5 h. The reaction mixture was evaporated
under vacuum, and the collected solid was crystallized
from the ethanol to yield 13a–c.
1-((4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)
3-((4-(2-(4-Chlorophenyl)-4-oxoquinazolin-3(4H)-yl)
phenoxy)methyl)quinazoline-2,4(1H,3H)-dione (11).
A
mixture of the azide derivative 8 (2.1 g; 5 mmol) and
anthranilic acid (0.68 g, 5 mmol) in dry dioxane
(20 mL) was refluxed for 5 h. The resulting solid was
collected and crystallized from ethanol to give
compound 11.
phenoxy)methyl)-3-phenylurea (13a).
Yellow crystals;
Yellow crystals; yield 65%; mp 123–124°C; IR (KBr,
yield 66%; mp 108–110°C; IR (KBr, cmꢀ1): 3466 (NH),
cmꢀ1): 3314 (NH), 1759, 1731, 1718 (C═O), 1655
1733, 1666 (C═O), 1655 (C═N); H NMR (DMSO-d6, δ
1
1
(C═N); H NMR (DMSO-d6, δ ppm): 4.20 (s, 2H, CH2),
ppm): 3.90 (s, 2H, CH2), 7.10–7.81 (m, 17H, Ar─H),
9.50 (br. s, 1H, NH, exchangeable with D2O), 9.80 (br. s,
1H, NH, exchangeable with D2O); MS (m/z, (relative
abundance, %)): 498 (M + 2, 6), 496 (M+, 20); Anal.
Calcd for C28H21ClN4O3: C, 67.67; H, 4.26; N, 11.27;
6.51–7.20 (m, 16H, Ar─H), 10.30 (br. s, 1H, NH,
exchangeable with D2O); MS (m/z, (relative abundance,
%)): 524 (M + 2, 8), 522 (M+, 25); Anal. Calcd for
C29H19ClN4O4: C, 66.61; H, 3.66; N, 10.71; Found: C,
66.58; H, 3.64; N, 10.75.
Found: C, 67.70; H, 4.30; N, 11.30.
1-(4-Bromophenyl)-3-((4-(2-(4-chlorophenyl)-4-
General procedures for the synthesis of 12a–c.
A
oxoquinazolin-3(4H)-yl)phenoxy)methyl)urea
(13b).
mixture of the azide derivatives 8 (2.1 g; 5 mmol) and
the appropriate phenol, namely, phenol, 4-aminophenol,
or 4-bromophenol (5 mmol), in dry benzene (20 mL) was
refluxed for 6 h. The reaction mixture was evaporated
under vacuum and petroleum ether was added. The
collected solid was crystallized from the benzene: ethanol
Yellowish red crystals; yield 75%; mp 149–150°C; IR
(KBr, cmꢀ1): 3445 (NH), 1728, 1665 (C═O), 1655
1
(C═N), 600 (C─Br); H NMR (DMSO-d6, δ ppm): 4.10
(s, 2H, CH2), 7.10–7.81 (m, 16H, Ar─H), 9.52 (br. s,
1H, NH, exchangeable with D2O), 9.89 (br. s, 1H, NH,
exchangeable with D2O); MS (m/z, (relative abundance,
%)): 579 (5), 577 (M + 2, 11), 575 (M+, 25); Anal. Calcd
for C28H20BrClN4O3: C, 58.40; H, 3.50; N, 9.73; Found:
(50:50) to provide 12a–c.
Phenyl((4-(2-(4-chlorophenyl)-4-oxoquinazolin-3(4H)-yl)
phenoxy)methyl)carbamate (12a).
Yellow crystals; yield
73%; mp 118–120°C; IR (KBr, cmꢀ1): 3344 (NH), 1726,
C, 58.46; H, 3.46; N, 9.76.
1-(4-Chlorophenyl)-3-((4-(2-(4-chlorophenyl)-4-
1
1679 (C═O), 1655 (C═N); H NMR (DMSO-d6, δ ppm):
oxoquinazolin-3(4H)-yl)phenoxy)methyl)urea
(13c).
4.40 (s, 2H, CH2), 7.10–7.81 (m, 17H, Ar─H), 10.80 (br.
s, 1H, NH, exchangeable with D2O); MS (m/z, (relative
abundance, %)): 497 (M+, 25); Anal. Calcd for
C28H20ClN3O4: C, 67.54; H, 4.05; N, 8.44; Found: C,
Yellowish red crystals; yield 79%; mp 173–174°C; IR
(KBr, cmꢀ1): 3404 (NH), 1727, 1665 (C═O), 1655
1
(C═N), 630 (C─Br); H NMR (DMSO-d6, δ ppm): 4.11
(s, 2H, CH2), 7.10–7.71 (m, 16H, Ar─H), 9.60 (br. s,
1H, NH, exchangeable with D2O), 9.93 (br. s, 1H, NH,
exchangeable with D2O); MS (m/z, (relative abundance,
%)): 532 (M + 2, 9), 530 (M+, 25); Anal. Calcd for
C28H20Cl2N4O3: C, 63.29; H, 3.79; N, 10.54; Found: C,
63.30; H, 3.80; N, 10.50.
67.51; H, 4.08; N, 8.41.
4-Aminophenyl((4-(2-(4-chlorophenyl)-4-oxoquinazolin-
3(4H)-yl)phenoxy)methyl)carbamate (12b). Yellow crystals;
yield 55%; mp 134–136°C; IR (KBr, cmꢀ1): 3415, 3355,
1
3225 (NH, NH2), 1716, 1699 (C═O), 1655 (C═N); H
NMR (DMSO-d6, δ ppm): 4.40 (s, 2H, CH2), 5.80 (br. s,
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2018
Synthesis and Anticancer Evaluation of Some New Quinazoline Derivatives
ANTIMICROBIAL ASSAY serum. Medium was changed at 48-h intervals. Cells
were dissociated with 0.25% trypsin. Experimental
cultures were plated in micrititer plates (Costar,
Cambridge, MA) containing 0.2 mL of growth medium
per well at densities of 1000–200,000 cells per well.
Cell fixation Cells attached to the plastic substratum
were fixed by gently laying 50 uL of cold 50%
trichloroacetic acid (TCA) (4°C) on top of the growth
medium in each well to produce a final TCA concentration
of 10%. The cultures were incubated at 4°C for 1 h and
then washed five times with tap water to remove TCA,
growth medium and low – molecular weight metabolites,
and serum protein. Plates were air dried and then stored
until use. Background optical densities were measured in
wells incubated with growth medium without cells.
Thirteen compounds were selected for the screening
from the synthesized compounds. All microorganisms
were kindly provided from culture collection of the
Regional Center for Mycology and Biotechnology, Cairo
University. The antimicrobial screening of some of the
synthesized compounds was carried out using Kirby–
Bauer disc diffusion method [16]. Briefly, 100 μL of the
tested bacteria/fungi was grown in 10 μL of fresh media
until they reached a count of approximately 108 cells per
mL for bacteria or 105 cells per mL for fungi [17,18]. A
total of 100 μL of microbial suspension was spread onto
agar plates corresponding to the broth in which they were
maintained. Plates inoculated with filamentous fungi as
Aspergillus Niger at 25°C for 48 h; Gram-positive bacteria,
as S. aureus; Gram-negative bacteria as E. coli, they were
incubated at 35–37°C for 24–48 h and yeast as C. albicans
incubated at 30°C for 24–48 h and then the diameters of
the inhibition zones were measured in millimeters.
Standard discs of Ampicillin (antibacterial agent) and
Amphotericin B (antifungal agent) were served as positive
controls for antimicrobial activity, but filter discs impreg-
nated with 10 μL of DMSO were used as a negative con-
trol. The agar used is Meuller–Hinton agar. Blank paper
discs (Schleicher & Schull, Spain) with a diameter
8.0 mm were impregnated 10 μL of tested concentration
of the stock solutions. When a filter paper disc impregnated
with a tested chemical is placed on agar, the chemical
will diffuse from the disc into the agar. This diffusion will
place the chemical in the agar only around the disc. The
solubility of the chemical and its molecular size will deter-
mine the size of the area of chemical infiltration around
the disc. If an organism is placed on the agar, it will not
grow in the area around the disc if it is susceptible to the
chemical, this area of no growth around the disc is known
as “zone of inhibition” of “clear zone.” Standard discs of
Tetracycline (antibacterial agent), Amphotericin B (anti-
fungal agent) served as positive controls for antimicrobial
activity, but filter discs impregnated with 10 μL of solvent
(distilled water, chloroform, DMSO) were used as a nega-
tive control.
Dying.
The anionic dye sulforhodamine B (SRB,
Sigma Chemical Co.) was dissolved in 1% acetic acid for
staining and extracted from cells with 10-mM unbuffered
Tris base [tris (hydroxymethyl)aminomethane].
Sulforhodamine assay. TCA-fixed cells were stained
for 30 min with 0.4% (w/v) SRB dissolved in 1% acetic
acid. At the end of the staining period, SRB was removed
and cultures were quickly rinsed four times with 1%
acetic acid to remove unbound dye. The acetic acid was
poured directly into permitted rinsing to be performed so
quickly that desorption of protein-bound dye did not
occur. Residual solution was removed by sharply flicking
plates over a sink, which ensured the complete removal
of rinsing solution. Because of the strong capillary action
in 96-well plates, draining by gravity alone often failed to
remove the rinsing solution when plates were simply
inverted.
After being rinsed, the cultures were air dried until no
standing moisture was visible. Bound dye was solubilized
with 10-mM unbuffered Tris base (pH 10.5) for 5 min on
gyratory shaker.
Optical density was read on a UVmax microtiter plate
reader (Molecular Devices, Menlo Park, CA) at 564 nm
for maximum sensitivity.
Acknowledgment. The authors thank the National Research
Centre for the financial support (Project number 11010317).
ANTICANCER SCREENING
REFERENCES AND NOTES
Ten compounds were selected for the anticancer
screening: Four concentrations were taken of each
compound: 1, 2.5, 5 and 10 μg/Ml Cells. Two human
cell lines were used in this experiment: (a) Human liver
carcinoma cell line (HepG2) and (b) human breast
carcinoma cell line (MCF7) [19]. Stock cultures were
grown in T-75 flasks containing 50 mL of RP Mi-1640
Medium with glutamine bicarbonate and 5% fetal calf
[1] Verhaeghe, P.; Azas, N.; Gasquet, M.; Hutter, S.; Ducros, C.;
Laget, M.; Rault, S.; Rathelot, P.; Vanelle, P. Bioorg Med Chem Lett
2008, 18, 396.
[2] Grover, G. S.; Kini, G. Eur J Med Chem 2006, 41, 256.
[3] Smits, R. A.; Lim, H. D.; Hanzer, A.; Zuiderveld, O. P.;
Guaita, E.; Adami, M.; Coruzzi, G.; Leurs, R.; de Esch, I. J. P. J Med
Chem 2008, 51, 2457.
[4] Georgey, H.; Abdel-Gawad, N.; Abbas, S. Molecules 2008,
13, 2557.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

R. S. Gouhar and M. M. Kamel
Vol 000
[5] Jain, K. S.; Bariwal, J. B.; Kathiravan, M. K.; Phoujdar, M. S.;
Sahne, R. S.; Chauhan, B. S.; Shah, A. K.; Yadav, M. R. Bioorg Med
Chem 2008, 16, 4759.
[6] Malamas, M. S.; Millen, J. J Med Chem 1991, 34, 1492.
[7] Shallal, H. M.; Russu, W. A. Eur J Med Chem 2011, 46, 2043.
[8] Chilin, A.; Conconi, M. T.; Marzaro, G.; Guiotto, A.; Urbani,
L.; Tonus, F.; Parnigotto, P. J Med Chem 2010, 53, 1862.
[9] Sagiv-Barfi, I.; Weiss, E.; Levitzki, A. Bioorg Med Chem
2010, 18, 6404.
[13] Wattanapiromsakul, C.; Forster, P. I.; Waterman, P. G. Phyto-
chemistry 2003, 64, 609.
[14] Amin, K. M.; Anwar, M. M.; Syam, Y. M.; Khedr, M.; Kamel,
M. M.; Kassem, E. M. Acta Pol Pharm Drug Res 2013, 70, 687.
[15] Amin, K. M.; Anwar, M. M.; Kamel, M. M.; Kassem, E. M.;
Syam, Y. M. Acta Pol Pharm Drug Res 2013, 70, 833.
[16] Bauer, A. W.; Kirby, W. M.; Sherris, C.; Turck, M. A. Am J
Clin Phathol 1996, 45, 493.
[17] Pfaller, M. A.; Burmeister, L.; Bartlett, M. A.; Rinaldi, M. G. J
Clin Microbiol 1988, 26, 1437.
[18] Ahamed, M. R.; Narren, S. F.; Sadiq, A. S. J Al-Nahrain Univ
2013, 16, 77.
[10] Fry, D. W.; Kraker, A. J.; McMichae, A.; Ambroso, L. A.;
Nelson, J. M.; Leopold, W. R.; Connors, R. W.; Bridges, A. J. Science
1994, 19, 265.
[11] Lewerenz, A.; Hentschel, S.; Vissiennon, Z.; Michael, S.;
Nieber, K. Drug Dev Res 2003, 58, 420.
[19] Skehan, p.; Storeng, R.; Scudiero, D. J Natl Cancer Inst 1990,
82, 1107.
[12] Malecki, N.; Carato, P.; Rigo, B.; Goossens, J.-F.; Houssin, R.;
Bailly, C.; He’nichart, J.-P. Bioorg Med Chem 2004, 12, 641.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet