Synthesis, anticancer and apoptosis-inducing activities of quinazoline–isatin conjugates: epidermal growth factor receptor-tyrosine kinase assay and molecular docking studies

Article abstract of DOI:10.1080/14756366.2017.1344981

A new series of quinazolinone compounds 16–34 incorporating isatin moieties was synthesized. The antitumor efficacy of the compounds against MDA-MB-231, a breast cancer cell line, and LOVO, a colon cancer cell line, was assessed. Compounds 20, 21, 22, 23, 25, 27, 28, 29, 30, 31, 32, 33, and 34 displayed potent antitumor activity against MDA-MB-231 and LOVO cells (IC₅₀: 10.38–38.67 μM and 9.91–15.77 μM, respectively); the comparative IC₅₀ values for 5-fluorouracil and erlotinib in these cells lines were 70.28 μM, 22.24 μM and 15.23 μM, 25.31 μM respectively. The EGFR-TK assay and induction of apoptosis for compound 31 were investigated to assess its potential cytotoxic activity as a representative example of the novel synthesized compounds. At a concentration of 10 μM, compound 31 exhibited efficient inhibitory effect against EGFR-TK and induced apoptosis in MDA-MB-231 cells. Furthermore, a molecular docking study for compound 31 and erlotinib was performed to verify the binding mode toward the EGFR kinase enzyme, and showed a similar interaction as that with erlotinib alone. Graphical Abstract: Compound 31 showed potent antitumor activity and efficient inhibitory effect against EGFR-TK and induced apoptosis of MDA-MB-231 cells at a concentration of 10 μM.

Full text of DOI:10.1080/14756366.2017.1344981

Journal of Enzyme Inhibition and Medicinal Chemistry
ISSN: 1475-6366 (Print) 1475-6374 (Online) Journal homepage: http://www.tandfonline.com/loi/ienz20
Synthesis, anticancer and apoptosis-inducing
activities of quinazoline–isatin conjugates:
epidermal growth factor receptor-tyrosine kinase
assay and molecular docking studies
Adel S. El-Azab, Abdullah Al-Dhfyan, Alaa A.-M. Abdel-Aziz, Laila A. Abou-
Zeid, Hamad M. Alkahtani, Abdulrahman M. Al-Obaid & Manal A. Al-Gendy
To cite this article: Adel S. El-Azab, Abdullah Al-Dhfyan, Alaa A.-M. Abdel-Aziz, Laila A. Abou-
Zeid, Hamad M. Alkahtani, Abdulrahman M. Al-Obaid & Manal A. Al-Gendy (2017) Synthesis,
anticancer and apoptosis-inducing activities of quinazoline–isatin conjugates: epidermal growth
factor receptor-tyrosine kinase assay and molecular docking studies, Journal of Enzyme Inhibition
and Medicinal Chemistry, 32:1, 935-944, DOI: 10.1080/14756366.2017.1344981
To link to this article: http://dx.doi.org/10.1080/14756366.2017.1344981
© 2017 The Author(s). Published by Informa
UK Limited, trading as Taylor & Francis
Group.
Published online: 18 Jul 2017.
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Date: 18 July 2017, At: 18:52

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY, 2017
VOL. 32, NO. 1, 935–944
https://doi.org/10.1080/14756366.2017.1344981
RESEARCH ARTICLE
Synthesis, anticancer and apoptosis-inducing activities of quinazoline–isatin
conjugates: epidermal growth factor receptor-tyrosine kinase assay and molecular
docking studies
Adel S. El-Azaba,b, Abdullah Al-Dhfyanc,d, Alaa A.-M. Abdel-Aziza,e, Laila A. Abou-Zeidf, Hamad M. Alkahtania,
Abdulrahman M. Al-Obaida and Manal A. Al-Gendya
b
aDepartment of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia; Department of Organic Chemistry,
c
Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt; Stem Cell & Tissue Reengineering Program, King Faisal Specialized Hospital and
d
Research Center, Riyadh, Saudi Arabia; Department of Pharmacology & Toxicology, Collage of Pharmacy, King Saud University, Riyadh, Saudi
e
f
Arabia; Department of Medicinal Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura, Egypt; Department of Pharmaceutical
Organic Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura, Egypt
ABSTRACT
ARTICLE HISTORY
Received 6 March 2017
Accepted 8 June 2017
A new series of quinazolinone compounds 16–34 incorporating isatin moieties was synthesized. The anti-
tumor efficacy of the compounds against MDA-MB-231, a breast cancer cell line, and LOVO, a colon cancer
cell line, was assessed. Compounds 20, 21, 22, 23, 25, 27, 28, 29, 30, 31, 32, 33, and 34 displayed potent
antitumor activity against MDA-MB-231 and LOVO cells (IC50: 10.38–38.67 lM and 9.91–15.77 lM, respect-
ively); the comparative IC50 values for 5-fluorouracil and erlotinib in these cells lines were 70.28 lM,
22.24 lM and 15.23 lM, 25.31 lM respectively. The EGFR-TK assay and induction of apoptosis for com-
pound 31 were investigated to assess its potential cytotoxic activity as a representative example of the
novel synthesized compounds. At a concentration of 10 lM, compound 31 exhibited efficient inhibitory
effect against EGFR-TK and induced apoptosis in MDA-MB-231 cells. Furthermore, a molecular docking
study for compound 31 and erlotinib was performed to verify the binding mode toward the EGFR kinase
enzyme, and showed a similar interaction as that with erlotinib alone.
KEYWORDS
Synthesis; isatin;
quinazolinone; antitumor;
EGFR-TK; molecular docking
Graphical Abstract: Compound 31 showed potent antitumor activity and efficient inhibitory effect against
EGFR-TK and induced apoptosis of MDA-MB-231 cells at a concentration of 10 lM.
Introduction
has broad spectrum antitumor activity with a mean GI50 value of
3.16 lM, in addition to EGFR-TK inhibitory activity11 (Figure 1).
Additionally, isatin derivatives exhibit broad spectrum bio-
logical effects such as anticancer activity26. A 5-fluoro-3-substi-
tuted isatin analog (Sunitinib) was approved by the FDA for the
Cancer is one of the most worldwide dangerous health problems
and is one of the leading causes of death1. Many of the current anti-
cancer agents are highly toxic and nonspecific, so the production of
innovative, safe, and selective anticancer molecules is an important
goal for the medicinal chemistry researchers. The quinazolinone treatment of renal carcinoma and gastrointestinal stromal
scaffold is a vital structure in medicinal chemistry2–22
.
tumors27,28 (Figure 1).
Anilinoquinazolines, such as gefitinib23,24 and erlotinib25, have
been established as EGFR kinase inhibitors for the treatment of
Methyl
3-(1-(4-bromobenzyl)-2,3-dioxoindolin-5-yl)acrylate
showed broad spectrum anticancer activity and a weak cytotoxic
breast cancer (Figure 1). The 3-phenethylquinazoline derivative (I) effect in normal human cells29.
A
series of indolinone
CONTACT Adel S. El-Azab
adelazab@ksu.edu.sa, adelazaba@yahoo.com; Alaa A.-M. Abdel-Aziz
almoenes@ksu.edu.sa, alaa_moenes@yahoo.com
Pharmaceutical Chemistry Department, College of Pharmacy, P.O. Box 2457, King Saud University, Riyadh-11451, Saudi Arabia
ß 2017 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distri-
bution, and reproduction in any medium, provided the original work is properly cited.

936
A. S. EL-AZAB ET AL.
hydrazides,
oxoindolin-3-ylidene)acetohydrazide (II) and 2-(4-fluoro-3-hydroxy-
phenyl)-N0-(2-oxoindolin-3-ylidene)acetohydrazide
(III), were
including
2-(6-oxo-1,6-dihydropyrimidin-4-yl)-N0-(2-
Materials and methods
Chemistry
reported as potent anticancer agents with IC50 values of 5.99 and
0.054 lM, respectively30 (Figure 1). As an attempt to develop
effective cytotoxic agents, we synthesized hybrids of quinazoline
conjugated to 5-substituted isatin that contained an acylhydra-
zone moiety and evaluated their cytotoxic activity. Additionally,
the EGFR-TK assay and apoptosis induction were investigated for
the most active compound, as a representative example of the
novel synthesized compounds, to identify their potential cytotoxic
activity. A molecular docking study was conducted to verify the
structural requirements of the antitumor activity of the target
molecules and to support the results of binding of the active
compounds to EGFR31.
Melting points were recorded on Barnstead 9100 Electrothermal
melting point apparatus (UK). IR spectra (KBr) were recorded on a
FT-IR Perkin-Elmer spectrometer (Perkin Elmer Inc., MA). Nuclear
magnetic resonance (1H and 13C NMR) spectra were recorded on
Bruker 500 or 700 MHz spectrometers (Zurich, Switzerland) using
DMSO-d6 as the solvent. Microanalytical data (C, H, and N) were
performed on a Perkin-Elmer 240 analyzer (Perkin Elmer Inc., MA)
and agreed with the proposed structures within 0.4% of the
theoretical values. Mass spectra were recorded on a Varian TQ
320 GC/MS/MS mass spectrometer (Varian, Palo Alto, CA). 2-[(3-
Substituted-4(3H)-quinazolinon-2-yl)thio]acetohydrazides
(11–15)
were prepared according to previously reported methods11,19,22
.
Figure 1. Reported and proposed quinazoline–isatin conjugates with antitumor and tyrosine kinase inhibitory activity.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
937
Synthesis
of
2-((3-substituted-4-oxo-3,4-dihydroquinazolin-2- 1H, J ¼ 4.5 Hz), 7.55–7.32 (m, 8H), 7.06–6.92 (m, 2H), 5.37 (d, 2H,
J ¼ 10.0 Hz), 4.65 (s, 1H), 4.41 (s, 1H), 2.41 (d, 3H, J ¼ 11.0 Hz); MS:
yl)thio)-N'-(2-oxoindolin-3-ylidene)acetohydrazides (16–34)
[m/z, 483].
An equimolar amount of the appropriate 2-[(3-substituted-4(3H)-qui-
nazolinon-2-yl)thio]acetohydrazide (11–15) and substituted isatin
was added to methanol (15 ml) containing glacial acetic acid
(0.2ml) and refluxed for 4–6 h. The reaction mixture was filtered
while hot; the solid obtained was washed with methanol and dried.
2-((3-Benzyl-6-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)thio)-N'-(5-
chloro-2-oxoindolin-3-ylidene)acetohydrazide (21)
Yield: 83%; mp: 246–247 oC; IR (KBr, cmꢀ1) ꢀ: 3456, 3163 (2NH),
1741, 1713, 1685 (3C¼O); 13C-NMR (176 MHz, DMSO-d6): d 21.2,
39.6, 47.4, 113.2, 116.8, 126.1, 126.4, 127.2, 127.9, 129.0, 136.1,
136.6, 141.6, 145.2, 155.7, 161.2; 1H-NMR (700 MHz, DMSO-d6): d
11.74 (s, 0.5H), 11.42 (s, 0.5H), 10.96 (s, 0.5H), 8.36 (s, 0.5H), 7.90 (s,
1H), 7.68–7.56 (m, 2H), 7.50–7.28 (m, 7H), 6.94 (d, 1H, J ¼ 5.5 Hz),
5.37 (s, 2H), 4.67 (s, 1.5H), 4.23–4.12 (m, 0.5H), 2.41 (s, 3H); MS: [m/
z, 517; M þ 2, 519].
2-((3-Benzyl-4-oxo-3,4-dihydroquinazolin-2-yl)thio)-N'-(2-oxoindo-
lin-3-ylidene)acetohydrazide (16)
Yield: 83%; mp: 250–251 oC; IR (KBr, cmꢀ1) ꢀ: 3421, 3160 (2NH),
1744, 1725, 1693 (3C¼O); 13C-NMR (176 MHz, DMSO-d6): d 29.5,
47.5, 111.6, 115.6, 119.1, 122.1, 123.1, 126.3, 126.6, 127.1, 127.2,
127.9, 129.0, 135.3, 135.4, 136.0, 142.9, 147.1, 156.9, 161.2, 163.0;
1H-NMR (700 MHz, DMSO-d6): d 11.54 (s, 0.5H), 11.33 (s, 0.5H),
10.86 (s, 0.5H), 8.14 (s, 0.5H), 8.11 (d, 1H, J ¼ 5.5 Hz), 7.75 (s, 1H),
7.57–7.46 (m, 2H), 7.41–7.28 (m, 6H), 7.08–7.03 (m, 1H), 6.93 (d, 1H,
J ¼ 5.5 Hz), 5.38 (s, 2H), 4.69 (s, 1H), 4.24 (s, 1H); MS: [m/z, 469].
2-((3-Benzyl-6-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)thio)-N'-(5-
fluoro-2-oxoindolin-3-ylidene)acetohydrazide (22)
Yield: 83%; mp: 272–274 oC; IR (KBr, cmꢀ1) ꢀ: 3448, 3182 (2NH),
1762, 1717, 1686 (3C¼O); 13C-NMR (125 MHz, DMSO-d6): d 20.6,
34.6, 46.9, 111.3, 112.2, 113.2, 113.4, 115.5, 115.6, 118.4, 125.6,
125.8, 126.7, 127.4, 128.5, 135.6, 135.8, 136.0, 138.7, 140.1, 144.7,
155.2, 156.5, 158.4, 160.7, 164.6; 1H-NMR (500 MHz, DMSO-d6): d
11.58 (s, 0.5H), 11.31 (s, 0.5H), 10.84 (s, 1H), 8.17 (d, 1H, J ¼ 8.0 Hz),
7.88 (s, 1H), 7.54 (dd, 1H, J ¼ 1.5 and 7.0 Hz), 7.36–7.25 (m, 7H),
6.93–6.90 (m, 1H), 5.37 (s, 2H), 4.65 (s, 1H), 4.59 (s, 1H), 2.39
(s, 3H); MS: [m/z, 501].
2-((3-Benzyl-4-oxo-3,4-dihydroquinazolin-2-yl)thio)-N'-(5-chloro-2-
oxoindolin-3-ylidene)acetohydrazide (17)
Yield: 83%; mp: 275–276 oC; IR (KBr, cmꢀ1) ꢀ: 3448, 3178 (2NH),
1723, 1718, 1695 (3C¼O); 13C NMR (176 MHz, DMSO-d6): d 29.4,
47.5, 113.1, 116.8, 119.1, 120.8, 121.9, 126.3, 126.6, 127.1, 127.2,
127.9, 129.0, 135.3, 135.4, 136.0, 141.6, 147.0, 147.1, 156.8, 161.2;
1H-NMR (700 MHz, DMSO-d6): d 11.77 (s, 0.5 H), 11.44 (s, 0.5 H),
10.98 (s, 0.5 H), 8.36 (s, 0.5 H), 8.10 (d, 1H, J ¼ 6.5 Hz), 7.74 (t, 1H,
J ¼ 5.5 Hz), 7.65–7.29 (m, 9H), 6.99–6.93 (m, 1H), 5.38 (s, 2H), 4.68
(s, 1H), 4.25 (s, 1H); MS: [m/z, 503; M þ 2, 505].
2-((3-Benzyl-6-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)thio)-N'-(5-
nitro-2-oxoindolin-3-ylidene)acetohydrazide (23)
Yield: 83%; mp: 292–294 oC; IR (KBr, cmꢀ1) ꢀ: 3467, 3167 (2NH),
1741, 1702, 1687 (3C¼O); 13C-NMR (176 MHz, DMSO-d6): d 21.1,
40.4, 47.48, 111.9, 115.5, 116.3, 118.9, 121.0, 122.0, 126.3, 126.4,
127.2, 127.9, 129.0, 136.1, 136.3, 136.6, 136.6, 142.5, 143.2, 145.1,
148.0, 155.7, 161.2, 165.4; 1H-NMR (700 MHz, DMSO-d6): d 12.56
(0.5H), 11.92 (0.5H), 11.53 (0.5H), 9.12 (0.5H), 8.29 (dd, 1H, J ¼ 5.5
and 15.0 Hz), 7.88 (s, 1H), 7.54 (d, 1H, J ¼ 5.5 Hz), 7.36–7.27 (m, 7H),
7.10 (d, 1H, J ¼ 6.0 Hz), 5.37 (s 2H), 4.60 (s, 1H), 4.27 (s, 1H), 2.40 (s,
3H); MS: [m/z, 528].
2-((3-Benzyl-4-oxo-3,4-dihydroquinazolin-2-yl)thio)-N'-(5-fluoro-2-
oxoindolin-3-ylidene)acetohydrazide (18)
Yield: 83%; mp: 244–245 oC; IR (KBr, cmꢀ1) ꢀ: 3410, 3169 (2NH),
1717, 1702, 1692 (3C¼O); 13C-NMR (176 MHz, DMSO-d6): d 29.3,
47.5, 108.4, 112.7, 118.5, 119.1, 121.5, 126.4, 126.7, 127.1, 127.2,
127.9, 129.0, 134.5, 135.4, 135.9, 139.2, 147.0, 156.2, 158.1, 159.4,
161.2, 163.0; 1H-NMR (700 MHz, DMSO-d6): d 13.48 (s, 0.4H), 12.74
(s, 0.6H), 11.38 (s, 1H), 8.13 7.36 (m, 12H), 5.40 (s, 2H), 4.70 (s, 1H),
4.28 (s, 1H); MS: [m/z, 487].
2-((4-Oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)thio)-N'-(2-oxoindo-
lin-3-ylidene)acetohydrazide (24)
2-((3-Benzyl-4-oxo-3,4-dihydroquinazolin-2-yl)thio)-N'-(5-nitro-2-
oxoindolin-3-ylidene)acetohydrazide (19)
Yield: 83%; mp: 304–305 oC; IR (KBr, cmꢀ1) ꢀ: 3449, 3223 (2NH),
1726, 1712, 1698 (3C¼O); 13C-NMR (125 MHz, CDCl3-DMSO-d6): d
34.6, 111.0, 119.4, 119.6, 120.538, 120.8, 122.4, 126.0, 126.4, 129.3,
129.4, 129.9, 131.5, 134.6, 135.4, 137.6, 142.4, 146.9, 155.7, 160.6,
Yield: 83%; mp: 313–315 oC; IR (KBr, cmꢀ1) ꢀ: 3467, 3279 (2NH),
1741, 1701, 1655 (3C¼O); 13C-NMR (176 MHz, DMSO-d6): d 47.5,
111.2, 111.8, 115.5, 119.1, 121.0, 122.1, 126.3, 126.6, 127.1, 127.2,
127.9, 128.0, 129.0, 135.3, 135.4, 136.0, 142.5, 143.2, 147.0, 148.1,
156.8, 161.2; 1H-NMR (700 MHz, DMSO-d6): d 12.27 (s, 0.5H), 11.93
(s, 0.5H), 11.56 (s, 0.5H), 9.12 (s, 0.5H), 8.34 (dd, 1H, J ¼ 8.5 Hz), 8.10
(d, 1H, J ¼ 8.0 Hz), 7.72 (t, 1H, J ¼ 7.5 and 8.0 Hz), 7.45 (t, 1H,
J ¼ 7.5 Hz), 7.37–7.27 (m, 6H), 7.11 (d, 1H, J ¼ 9.0 Hz), 5.38 (s, 2H),
4.66 (s, 1H), 4.30 (s, 1H); MS: [m/z, 514].
1
162.4, 164.9; H NMR (500 MHz, DMSO-d6): d 13.49 (s, 0.56H), 12.72
(s, 0.46H), 11.26 (s, 1H), 8.07 (dd, 1H, J ¼ 1.0 and 8.0Hz), 7.74 (s, 1H),
7.60–7.42 (m, 8H), 7.33 (t, 1H, J ¼ 8.0 Hz), 7.0526 (d, 1H, J ¼ 6.0 Hz),
6.95–6.87 (m, 1H), 4.55 (s, 1H), 4.08 (s, 1H); MS: [m/z, 455].
N'-(5-chloro-2-oxoindolin-3-ylidene)-2-((4-oxo-3-phenyl-3,4-dihydro-
quinazolin-2-yl)thio)acetohydrazide (25)
Yield: 83%; mp: 328–329 oC; IR (KBr, cmꢀ1) ꢀ: 3447, 3259 (2NH),
1730, 1702, 1659 (3C¼O); 13C-NMR (176 MHz, DMSO-d6): d 21.2,
2-((3-Benzyl-6-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)thio)-N'-(2-
oxoindolin-3-ylidene)acetohydrazide (20)
Yield: 83%; mp: 270–271 oC; IR (KBr, cmꢀ1) ꢀ: 3412, 3273 (2NH), 47.4, 47.4, 113.2, 116.8, 126.1, 126.4, 127.2, 127.9, 129.0, 136.1,
1793, 1724, 1686 (3C¼O); 13C-NMR (125 MHz, DMSO-d6): d 20.7, 136.6, 141.6, 145.2, 155.7, 161.2; 1H-NMR (700 MHz, DMSO-d6): d
39.9, 46.9, 110.6, 115.2, 118.4, 121.6, 125.9, 126.7, 127.4, 128.5, 13.42 (s, 0.5H), 12.63 (s, 0.5H), 11.43 (s, 1H), 8.07 (d, 1H, J ¼ 5.5 Hz),
135.6, 136.0, 144.7, 155.2, 160.7; 1H-NMR (500 MHz, DMSO-d6): d 7.79 (s, 1H), 7.72–7.61 (m, 4H), 7.51–7.42 (m, 5H), 6.99–6.93 (m,
11.51–11.32 (m, 1H), 10.84 (d, 1H, J ¼ 7.0 Hz), 8.13 (s, 1H), 7.88 (d, 1H), 4.57 (s, 1H), 4.12 (s, 1H); MS: [m/z, 489; M þ 2, 491].

938
A. S. EL-AZAB ET AL.
N'-(5-fluoro-2-oxoindolin-3-ylidene)-2-((4-oxo-3-phenyl-3,4-dihydro-
quinazolin-2-yl)thio)acetohydrazide (26)
7.87 (s, 1H), 7.60–7.11 (m, 9H), 4.59 (s, 1H), 4.35 (s, 0.3H), 4.14 (s,
0.7H), 2.41 (s, 1H); MS: [m/z, 514].
Yield: 83%; mp: 310–312 oC; IR (KBr, cmꢀ1) ꢀ: 3429, 3256 (2NH),
1733, 1709, 1686 (3C¼O); 13C-NMR (125 MHz, DMSO-d6): d 34.7,
108.0, 112.2, 118.1, 119.4, 120.9, 126.0, 126.5, 129.4, 129.5, 130.0,
134.8, 135.6, 138.69 146.9, 157.3, 159.2, 160.5, 162.6; 1H-NMR
(500 MHz, DMSO-d6): d 13.47 (s, 0.5H), 12.69 (s, 0.5 H), 11.34 (s, 1H),
8.06 (d, 1H, J ¼ 8.0 Hz), 7.77 (s, 1H), 7.61–7.34 (m, 8H), 7.20 (s, 1H)
6.95– 6.90 (m, 1H), 4.56 (s, 1H), 4.11 (s, 1H); MS: [m/z, 473].
2-((4-Oxo-3-phenethyl-3,4-dihydroquinazolin-2-yl)thio)-N'-(2-oxoin-
dolin-3-ylidene)acetohydrazide (32)
Yield: 83%; mp: 273–274 oC; IR (KBr, cmꢀ1) ꢀ: 3448, 3133 (2NH),
1715, 1686, 1636 (3C¼O); 13C-NMR (176 MHz, DMSO-d6): d 33.9,
40.4, 46.0, 111.1, 111.5, 119.2, 120.2, 122.1, 123.1, 126.2, 126.5,
126.9, 127.2, 129.1, 135.1, 135.2, 138.1, 142.9, 147.0, 147.0, 160.8,
160.8; 1H-NMR (700 MHz, DMSO-d6): d 11.55 (s, 0.5H), 11.28, (s,
0.5H), 10.83 (s, 0.5H), 8.16 (s, 0.5H), 8.15 (d, 1H, J ¼ 2.0 Hz), 8.07 (d,
1H, J ¼ 5.5 Hz), 7.80 (d, 1H, J ¼ 6.0 Hz), 7.68–7.26 (m, 8H), 7.10–7.00
(m, 1H), 6.93–6.90 (m, 1H), 4.74–4.49 (m, 1.5H), 4.41–4.36 (m, 2.5H,
J ¼ 5.0 and 7.5 Hz), 3.07–3.00 (m, 2H); MS: [m/z, 483].
N'-(5-nitro-2-oxoindolin-3-ylidene)-2-((4-oxo-3-phenyl-3,4-dihydro-
quinazolin-2-yl)thio)acetohydrazide (27)
Yield: 83%; mp: 337–338 oC; IR (KBr, cmꢀ1) ꢀ: 3431, 3188 (2NH),
1
1730, 1712, 1691 (3C¼O); H-NMR (500 MHz, DMSO-d6): d 13.31 (s,
0.5 H), 12.53 (s, 0.5), 11.94 (s, 1H), 8.29 (d, 2H, J ¼ 6.5 Hz), 8.00 (d,
1H, J ¼ 7.5 Hz), 7.78 (s, 1H), 7.61–7.47 (m, 8H), 7.13 (s, 1H), 4.61 (s,
1H), 4.16 (s, 1H); MS: [m/z, 500].
N'-(5-chloro-2-oxoindolin-3-ylidene)-2-((4-oxo-3-phenethyl-3,4-dihy-
droquinazolin-2-yl)thio)acetohydrazide (33)
Yield: 83%; mp: 233–235 oC; IR (KBr, cmꢀ1) ꢀ: 3469, 3167 (2NH),
1710, 1676, 1646 (3C¼O); 13C-NMR (176 MHz, DMSO-d6): d 33.9,
40.4, 46.0, 112.4, 116.8, 119.2, 121.9, 126.1, 126.2, 126.5, 126.9,
127.2, 129.1, 135.1, 135.2, 138.1, 143.0, 147.0, 156.2, 160.8, 164.8,
172.4; 1H-NMR (700 MHz, DMSO-d6): d 11.67 (s, 0.5H), 11.58 (s,
0.5H), 10.97 (s, 1H), 8.37 (s, 1H), 8.06 (d, 1H, J ¼ 5.5 Hz), 7.70 (t, 1H,
J ¼ 5.0 Hz), 7.44–7.21 (m, 8H), 6.93 (d, 1H, J ¼ 6.0 Hz), 4.70 (s, 1H),
4.52 (s, 1H), 4.29 (t, 2H, J ¼ 5.5 Hz), 3.05 (t, 2H, J ¼ 5.5 and 5.5 Hz);
MS: [m/z, 517; M þ 2, 519].
2-((6-Methyl-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)thio)-N'-(2-
oxoindolin-3-ylidene)acetohydrazide (28)
Yield: 83%; mp: 305–306 oC; IR (KBr, cmꢀ1) ꢀ: 3421, 3298 (2NH),
1725, 1695, 1652 (3C¼O); 13C-NMR (176 MHz, DMSO-d6): d 21.1,
35.1, 111.6, 115.7, 119.7, 120.1, 121.3, 122.1, 123.1, 126.2, 126.3,
129.9, 130.0, 130.4, 136.3, 136.4, 136.5, 136.6, 142.9, 144.3, 145.6,
161.1, 163.0, 165.0; 1H-NMR (700 MHz, DMSO-d6): d 11.46 (s, 0.5H),
11.31 (s, 0.5H), 10.85 (s, 0.5H), 8.15 (s, 0.5H), 7.86 (s, 1H), 7.63–7.48
(m, 7H), 7.40–7.35 (m, 2H), 7.05 (t, 1H, J ¼ 5.0 and 5.5 Hz),
6.97–6.90 (m, 1H), 4.55 (s, 1H), 4.28 (s, 0.5H), 4.08 (s, 0.5 H), 2.40 (s,
3H); MS: [m/z, 469].
N'-(5-fluoro-2-oxoindolin-3-ylidene)-2-((4-oxo-3-phenethyl-3,4-dihy-
droquinazolin-2-yl)thio)acetohydrazide (34)
Yield: 83%; mp: 257–258 oC; IR (KBr, cmꢀ1) ꢀ: 3442, 3267 (2NH),
1
1719, 1683, 1639 (3C¼O); H-NMR (500 MHz, DMSO-d6): d 11.61 (s,
N'-(5-chloro-2-oxoindolin-3-ylidene)-2-((6-methyl-4-oxo-3-phenyl-
3,4-dihydroquinazolin-2-yl)thio)acetohydrazide (29)
0.5H), 11.30 (s, 0.5H), 10.85 (s, 0.5H), 8.19 (d, 0.5H, J ¼ 8.5 Hz), 8.05
(d, 1H, J ¼ 8.0 Hz), 7.70 (t, 1H, J ¼ 7.0 and 7.5 Hz), 7.50–7.21 (m, 9H),
6.91 (dd, 1H, J ¼ 4.0 and 4.5 Hz), 4.73 (s, 1H), 4.65 (s, 1H), 4.28 (dd,
2H, J ¼ 4.0 Hz), 3.05 (t, 2H, J ¼ 7.5 Hz); MS: [m/z, 501].
Yield: 83%; mp: 328–330 oC; IR (KBr, cmꢀ1) ꢀ: 3419, 3149 (2NH),
1
1721, 1689, 1646 (3C¼O); H-NMR (500 MHz, DMSO-d6): d 11.70 (s,
0.5H), 11.44 (s, 0.5H), 10.95 (s, 1H), 8.35 (s, 1H), 7.87 (s, 1H),
7.60–7.49 (m, 7H), 7.36 (s, 1H), 6.93 (d, 1H, J ¼ 8.0 Hz), 4.51 (s, 1H),
4.35 (s, 0.75H), 4.10 (s, 0.25H), 2.42 (s, 3H); MS: [m/z, 503; M þ 2,
505].
Biology
WST-1 cell proliferation assay
N'-(5-fluoro-2-oxoindolin-3-ylidene)-2-((6-methyl-4-oxo-3-phenyl-
3,4-dihydroquinazolin-2-yl)thio)acetohydrazide (30)
The cell proliferation assay was conducted according to a previ-
ously reported method32.
Yield: 83%; mp: 281–282 oC; IR (KBr, cmꢀ1) ꢀ: 3448, 3283 (2NH),
1725, 1699, 1662 (3C¼O); 13C-NMR (125 MHz, DMSO-d6): d 20.6,
34.6, 111.3, 112.2, 119.2, 121.0, 125.7, 125.8, 129.4, 129.5, 129.9,
135.6, 135.8, 136.0, 136.1, 138.6, 145.0, 145.1, 155.6, 156.5,
157.3, 159.2, 160.5, 162.6, 164.6; 1H-NMR (500 MHz, DMSO-d6): d
11.57 (s, 0.4H), 11.33 (s, 0.6H), 10.83 (s, 0.4H), 8.14 (s, 0.6H),
7.86 (s, 1H), 7.61–7.49 (m, 7H), 7.35 (s, 1H), 7.26–7.21 (m, 1H),
6.91 (t, 1H, J ¼ 4.0 Hz), 4.55 (s, 1.4H), 4.09 (s, 0.6H), 2.41 (s, 3H);
MS: [m/z, 487].
Immunofluorescence microscopy
The EGFR immunofluorescence assay was conducted according to
a previously reported method33.
Apoptosis assay
Vybrant apoptosis assay kit (Annexin-V, APC conjugate; Molecular
ProbesTM) was used to evaluate cell viability in accordance with
the manufacturer’s recommendation33.
2-((6-Methyl-4-oxo-3-phenyl-3,4-dihydroquinazolin-2-yl)thio)-N'-(5-
nitro-2-oxoindolin-3-ylidene)acetohydrazide (31)
Docking methodology
Yield: 83%; mp: 344–345 oC; IR (KBr, cmꢀ1) ꢀ: 3446, 3196 (2NH),
1
1744, 1707, 1648 (3C¼O); H-NMR (700 MHz, DMSO-d6): d 13.29 (s, All modeling experiments were conducted with MOE programs
0.7H), 12.52 (s, 0.3H), 11.92 (s, 0.7H), 11.52 (s, 0.3H), 8.30 (s, 1H), running on a PC34. Hydrogen bonds with a bond length of up to

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
939
3.5 Å were considered. The starting coordinates of the X-ray crystal NH group of amides, and 1793–1713 cm–1 and 1676–1725 cm–1,
structure of the EGFR enzyme in complex with erlotinib (PDB owing to the presence of two C¼O groups in addition to the C¼O
code: 1M17) were obtained from the RCSB Protein Data Bank of of the 4-quinazolinone nucleus at 1698–1636 cm–1.
Brookhaven National Laboratory35. All hydrogens were added and
the enzyme structure was subjected to a refinement protocol in
Biological activity
which the constraints on the enzyme were gradually removed and
minimized until the RMS gradient was 0.01 kcal/mol Å. The energy
minimization was conducted using the AMBER molecular mechan-
ics force field. The lowest energy conformer, the “global-minima,”
was pre-positioned using the crystal structure ligand “erlotinib” as
a template at the enzyme-binding pocket.
Cell proliferation inhibition assay
The in vitro antitumor activity of compounds 16–34 against the
human breast cancer cell line, MDA-MB-231, and the colon cancer
cell line, LOVO, was determined by WST-1 assay32 using 5-FU and
erlotinib as a reference drugs, and IC50 was calculated for each
cell line (Table 1). In the present study, the active compounds
exhibited a characteristic selectivity potential in addition to broad-
spectrum antitumor activity.
For the selectivity against the MDA-MB-231 cell line, com-
pounds 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 30, 31, 32,
33, and 34 showed high activity (IC50: 10.38–20.21 lM); the com-
parative IC50 values for 5-FU and erlotinib were 70.28 and
22.24 lM respectively. On the other hand, compounds 28 and 29
(IC50: 37.41 and 38.67 lM); were less active than erlotinib but more
active than 5-FU.
Moreover, the LOVO cell line was sensitive toward compounds
19, 20, 21, 22, 23, 25, 27, 28, 29, 30, 31, 32, 33, and 34 (IC50:
9.91–17.53 lM); the comparative IC50 value for 5-FU and erlotinib
were 15.23 and 25.31 lM respectively. Compounds 17, 18, 24, and
26 were less active than 5-FU with IC50 values of 20.39–23.98 lM
but more active than erlotinib.
With regards to broad-spectrum antitumor activity, com-
pounds 20, 21, 22, 23, 25, 27, 30, 31, 32, 33, and 34 showed
strong antitumor activities against MDA-MB-231 cells and
LOVO cells, which was supported by the IC50 values
(10.38–20.21 lM and 9.91–15.77 lM, respectively). Moreover, com-
pound 31 showed the highest potency toward MDA-MB-231
cells and LOVO cells with IC50 values of 10.38 and 9.91 lM,
respectively.
Results and discussion
Chemistry
2-Mercapto-3-substituted-4(3H)-quinazolinones (1–5) were pre-
pared by heating anthranilic acid derivatives with an appropriate
isothiocyanate in ethanol containing a catalytic amount of triethyl-
amine. Accordingly, 2-[(3-substituted-4(3H)-quinazolinon-2-yl)th-
io]acetohydrazides (11–15) were obtained by stirring compounds
1–5 with ethyl 2-bromoacetate in acetone to yield the correspond-
ing ethyl 2-[(3-substituted-4(3H)-quinazolinon-2-yl)thio]acetates
(6–10), which were then stirred with hydrazine hydrate in etha-
nol11,19,22 (Scheme 1).
The 2-[(3-substituted-4-quinazolinon-2-yl)thio]-N'-(2-oxoindolin-
3-ylidene)acetohydrazides (16–34) were produced at 80–85% yield
by heating an appropriate 2-[(3-substituted-4(3H)-quinazolinon-2-yl)
thio]acetohydrazide (11–15) and isatin derivative in methanol con-
taining a catalytic amount of acetic acid26 (Scheme 2).
1H-NMR of compounds 16–34 revealed singlet signals corre-
sponding to the two NH groups at 13.48–10.85 and
11.94–8.13 ppm, in addition to presence of signals for SCH2CO at
4.79–4.05 ppm as a mixture of the E/Z isomers. Additionally, the IR
spectra of compounds 16–34 showed new bands at
3467–3410 cm–1 and 3298–3133 cm–1, which corresponded to the
OH
O
X
S
X
R
O
R
C
N
+
N
EtOH, TEA
reflux
NH2
N
SH
X= H, CH3
R= Bn, Ph, PhEt
1- X= H, R=Bn
- X=CH3, R=Bn
2
3- X=H, R=Ph
4- X=CH3, R=Ph
- X=H, R=PhEt
BrCH2COOEt
acetone, K2CO3
5
O
O
N
X
R
X
R
N
N
NH2NH2,
EtOH, r.t.
NH
O
S
N
S
NH2
O
O
11
- X= H R=Bn
12- X=CH3 R=Bn
6
- X= H R=Bn
7- X=CH3 R=Bn
13- X=H
- X=CH3 R=Ph
15- X=H
R=Ph
8- X=H
R=Ph
14
9
- X=CH3 R=Ph
R=PhEt
10-X=H
R=PhEt
Scheme 1. Synthesis of 2-[(3-substituted-4(3H)-quinazolinon-2-yl)thio]acetohydrazides 11–15.

940
A. S. EL-AZAB ET AL.
O
N
O
O
N
Y
O
X
R
S
X
R
S
NH
N
N
O
H
H
+
MeOH, AcOH
reflux
N
N
N
NH2
N
H
O
O
Y
16- X= H, R=Bn,
17- X=H, R=Bn,
1 - X=H, R=Bn,
Y=H
Y=Cl
Y=F
Y=H, Cl, F, NO2
11- X= H, R=Bn
8
12
13
- X=CH3, R=Bn
- X=H, R=Ph
19- X=H, R=Bn,
20- X=CH3, R=Bn,
Y=NO2
Y= H
Y= Cl
Y= F
Y= NO2
Y=H
Y=Cl
Y=F
Y=NO2
Y=H
14- X=CH3, R=Ph
15
21
- X=CH3, R=Bn,
- X=H, R=PhEt
22- X=CH3, R=Bn,
23- X=CH3, R=Bn,
24
- X= H, R=Ph,
25- X= H, R=Ph,
26- X= H, R=Ph,
27
- X= H, R=Ph,
28- X=CH3, R=Ph,
29- X=CH3, R=Ph,
30
- X=CH3, R=Ph,
31- X=CH3, R=Ph,
Y=Cl
Y=F
Y=NO2
32- X=H, R=PhEt, Y=H
33
- X=H, R=PhEt, Y=Cl
34- X=H, R=PhEt, Y=F
Scheme 2. Synthesis of quinazoline-isatin conjugates 16–34.
Table 1. In vitro antitumor activity of the newly synthesized compounds 16–34.
biparametric cytofluorimetric analysis32. After treatment with com-
pound 31 (10 lM for 24 h), the MDA-MB-231 breast cancer cells
were stained with DAPI and annexin V, and analyzed by flow
cytometry (Figure 3). Compound 31 was able to induce apoptosis
in MDA-MB-231 cells. Compound 31 induced apoptosis by a 30-
fold increase in the percentage of fluorescein isothiocyanate
annexin V (Annexin V-FITC)-positive apoptotic cells (right panel) in
comparison with untreated cells (left panel). Compound 31
increased the percentage of apoptotic cells by 5.6% and late
apoptotic cells by 61.4% compared with 1.3% and 2.6% in
untreated control cells, respectively. Moreover, the tested com-
pound induced necrosis in treated cells by 8.3% compared with
0.2% in untreated control cells.
Compounds
MDA-MB-231a IC50 (lM)c
LOVOb IC50 (lM)c
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
16.23 0.32
14.97 0.37
12.38 0.3
12.31 0.11
16.82 0.13
14.48 0.03
18.33 0.01
17.14 0.01
11.50 0.36
11.41 0.07
11.80 0.02
18.05 0.04
37.41 0.06
38.67 0.04
13.77 0.4
10.38 0.22
18.35 0.14
20.21 0.05
20.06 0.11
70.28 0.2
22.24 0.22
33.97 0.26
23.98 0.06
21.46 0.13
17.53 0.04
14.80 0.1
14.21 0.06
14.14 0.06
13.39 0.23
20.39 0.02
12.00 0.05
23.62 0.01
12.80 0.03
14.20 0.09
14.00 1.02
14.12 0.06
9.91 0.12
32
33
34
5-FU
Erlotinib
16.51 0.15
14.37 0.46
15.77 0.16
15.23 0.09
25.31 0.12
Structure–activity relationships
The structure–activity relationships of the tested compounds
revealed that 5-methyl-3-benzyl derivatives 20–23 (IC50:
14.48–18.33 lM and 13.39–14.80 lM) and 5-methyl-3-phenyl deriv-
atives 28–31 (IC50: 10.38–38.67 lM and 9.91–14.20 lM) showed sig-
nificant inhibition of MDA-MB-231 cells and LOVO cells, compared
with 5-FU (IC50: 70.28 lM and 15.23 lM), respectively (Table 1).
Moreover, unsubstituted 3-benzyl derivatives 16–19 (IC50:
12.31–16.23 lM and 17.53–33.97 lM), 3-unsubstituted phenyl deriv-
atives 24–27 (IC50: 11.41–18.05 and 12.0–23.62 lM) and unsubsti-
tuted 3-phenethyl derivatives 32–34 (IC50: 18.35–20.21 and
14.37–17.87 lM) were more selective for MDA-MB-231 cells than
LOVO colon cells, compared with 5-FU (IC50: 70.28 lM and
15.23 lM), respectively (Table 1).
In MDA-MB-231 cells, the unsubstituted 3-benzyl derivatives
16–19 (IC50: 12.31–16.23 lM) and unsubstituted 3-phenyl deriva-
tives 24–27 (IC50: 11.41–18.05 lM) were more active than the 5-
methyl-3-benzyl derivatives 20–23 (IC50: 14.48–18.33 lM) and 5-
methyl-3-phenyl derivatives 28–31 (IC50: 10.38–38.67 lM) respect-
ively. In the LOVO cells, the 5-methyl-3-benzyl derivatives 20–23
(IC50: 13.39–14.80 lM) and 5-methyl-3-phenyl derivatives 28–31
aAggressive human MDA-MB-231 (representative triple negative breast cancer
cells with high metastasis potential).
bAggressive human LOVO colon cell line (type IV metastasized colon cancer).
cIC50: concentration of the compound (lM) that produced 50% inhibition of cell
growth inhibition after 48 h of treatment.
GFR tyrosine kinase enzyme inhibition assay
The enzyme activity assay of the most active compound 31
toward the MDA-MB-231 breast cancer cell line was selected as
representative example of the compounds and administered at a
single concentration (10 lM) against EGFR-TK to investigate the
mechanism of action of the newly synthesized compounds33. The
immunofluorescence staining of EGFR in MDA-MB-231 cells treated
with compound 31 at 10 lM indicated a good selectivity of com-
pound 31 to EGFR-TK, as shown by inhibition of the level of EGFR
on the cell membrane as well as in the nucleus (Figure 2).
Apoptosis detection by flow cytometry
The effect of compound 31 on the apoptosis was investigated (IC50: 9.91–14.2 lM) were more active than the unsubstituted 3-
using DAPI (4,6-diamidino-2-phenylindole) and annexin V-FITC benzyl derivatives 16–19 (IC50: 17.53–33.97 lM) and unsubstituted

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
941
Figure 2. EGFR (left panel; green color) of MDA-MB-231 breast cell line and (right panel) MDA-MB-231 breast cell line after treatment with compound 31.
Figure 3. MDA-MB-231 breast cancer cell line was treated with compound 31 (right panel), which displayed an increased percentage of fluorescein isothiocyanate
annexin V (Annexin V–FITC), and untreated control cells (left panel).
3-phenyl derivatives 24–27 (IC50: 12.0–23.62 lM), respectively bonding with Leu768, while the second semicarbazide nitrogen
(Table 1).
atom performed hydrophilic interaction by cross interaction with
Pro717 through the water molecule in the pocket. The two adja-
cent conserved amino acids Leu768 and Met769 firmly held the
backbone of compound 31, which augmented the recognition
and the overall inhibition activity (Figure 4).
In contrast, compound 28 was bound in different manner,
which dramatically lowered the overall complementarity. Although
N1 was clearly recognized with hydrogen bonding to the distinct-
ive residue Met769, N3 was buried away from the surrounding
amino acids owing to the rigidity of the connected phenyl group.
However, the semicarbazide linkage enriched the hydrophilic inter-
action by cross interaction with Pro717 through the water molecule
in the pocket (Figure 5).
Molecular docking results
The antitumor activities of the weakly active compound 28 and
the highly active compound 31 in MDA-MB-23 cells, which highly
express epidermal growth factor receptor (EGFR)7,10,11,15,19,22 and
the binding activity of compound 31 with EGFR, encouraged us to
conduct molecular docking simulations of the binding site of the
EGFR kinase.
Compounds 28 and 31 were docked into the receptor active
site of EGFR along with their inhibitor erlotinib (TarcevaTM) (PDB
code: 1M17)35. All calculations were performed using MOE 2008.10
software34. The docking study of the most active compound 31
revealed that the quinazoline ring typically overlaid the corre-
sponding ring of erlotinib without clashing with the surrounding
amino acids. The substituted linkage at the C-2 hybrid of the bind-
ing of compound 31 in both the activation and catalytic loops
Conclusions
A new series of quinazolinone-isatin conjugates 16–34, which
strongly inhibited growth in the MDA-MB-231 breast cancer cell
where N1 was uniquely bound with the distinctive residue Met769
.
A semicarbazide nitrogen atoms was recognized via hydrogen line and LOVO colon cancer cell line, was synthesized. Compounds

942
A. S. EL-AZAB ET AL.
Figure 4. Docking of compound 31 (left panel) and superposition with erlotinib (right panel) in the receptor pocket of EGFR kinase. Compound 31 and erlotinib are
shown in green and cyan, respectively.
Figure 5. Docking of compound 28 (left panel) and superposition with erlotinib (right panel) in the receptor pocket of EGFR kinase. Compound 28 and erlotinib are
shown in yellow and cyan, respectively.
16–34 showed high activity against the human MDA-MB-231 of quinazolinone–isatin conjugates at
a single concentration
breast cell line (IC50: 10.38–38.67 lM) in comparison with 5-FU and (10 lM). Compound 31 increased the number of apoptotic cells by
erlotinib (IC50: 70.28 lM and 22.24 lM, respectively). Similarly, com- 5.6% and late apoptotic cells by 61.4% compared with 1.3 and
pounds 19–23, 25, and 27–34 possessed strong activity against 2.6%, respectively, in untreated control cells. Additionally, com-
the LOVO colon cancer cell line (IC50: 9.91–17.87 lM) in compari- pound 31 induced necrosis in treated cells by 8.3% compared
son with 5-FU and erlotinib (IC50: 15.23 lM and 25.31 lM, respect- with 0.2% in untreated control cells. A molecular docking simula-
ively). Compounds 20–23, 25, and 27–34 showed potent tion was performed for compounds 31 and 28 into the binding
antitumor activity against the MDA-MB-231 and LOVO cell lines site of EGFR kinase, which showed a similar binding mode to erlo-
(IC50: 10.38–38.67 lM and 9.91–15.77 lM, respectively. Compound tinib. The results of molecular docking can help in the design of
31 inhibited the level of EGFR-TK in the cell membrane, as well as new molecules with potential antitumor activity and good binding
in the nucleus, of MDA-MB-231 cells as a representative example to the enzyme receptor site.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
943
13. El-Azab AS. Synthesis of some new substituted 2-mercapto-
quinazoline analogs as potential antimicrobial agents.
Phosphorus Sulfur Silicon Relat Elem 2007;182:333–48.
14. El-Azab AS, Abdel-Hamide SG, Sayed-Ahmed MM, et al.
Novel 4 (3H)-quinazolinone analogs: synthesis and anticon-
vulsant activity. Med Chem Res 2013;22:2815–27.
15. El-Azab AS, Al-Omar MA, Alaa A-M, et al. Design, synthesis
and biological evaluation of novel quinazoline derivatives as
potential antitumor agents: molecular docking study. Eur J
Med Chem 2010;45:4188–98.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
The authors extend their appreciation to the Deanship of Scientific
Research at King Saud University for funding the work through
the research group project No. RG-1435-046.
16. El-Azab AS, ElTahir KE. Synthesis and anticonvulsant evalu-
ation of some new 2,3,8-trisubstituted-4 (3H)-quinazoline
derivatives. Bioorg Med Chem Lett 2012;22:327–33.
17. El-Azab AS, ElTahir KE. Design and synthesis of novel 7-ami-
noquinazoline derivatives: antitumor and anticonvulsant
activities. Bioorg Med Chem Lett 2012;22:1879–85.
18. El-Azab AS, ElTahir KE, Attia SM. Synthesis and anticonvul-
sant evaluation of some novel 4 (3H)-quinazolinones. Chem
Monthly 2011;142:837–48.
References
1. Senwar KR, Reddy TS, Thummuri D, et al. Design, synthesis
and apoptosis inducing effect of novel (Z)-3-(30-methoxy-40-
(2-amino-2-oxoethoxy)-benzylidene) indolin-2-ones as poten-
tial antitumour agents. Eur J Med Chem 2016;118:34–46.
2. Alaa A-M, Abou-Zeid LA, ElTahir KEH, et al. Design, synthesis
of 2, 3-disubstitued 4 (3H)-quinazolinone derivatives as anti-
inflammatory and analgesic agents: COX-1/2 inhibitory activ-
ities and molecular docking studies. Bioorg Med Chem
2016;24:3818–28.
19. Mohamed MA, Ayyad RR, Shawer TZ, et al. Synthesis and
antitumor evaluation of trimethoxyanilides based on
4
(3H)-quinazolinone scaffolds. Eur J Med Chem 2016;112:
106–13.
3. Alaa A-M, El-Azab AS, Abou-Zeid LA, et al. Synthesis, anti-
inflammatory, analgesic and COX-1/2 inhibition activities of
anilides based on 5, 5-diphenylimidazolidine-2, 4-dione scaf-
20. Alafeefy AM, Kadi AA, El-Azab AS, et al. Synthesis,
analgesic and anti-inflammatory evaluation of some new 3H-
quinazolin-4-one derivatives. Archiv Der Pharmazie 2008;
341:377–85.
21. Alaa A-M, Abou-Zeid LA, ElTahir KEH, et al. Synthesis, anti-
inflammatory, analgesic, COX-1/2 inhibitory activities and
molecular docking studies of substituted 2-mercapto-4 (3H)-
quinazolinones. Eur J Med Chem 2016;121:410–21.
22. Alanazi AM, Abdel-Aziz AA, Shawer TZ, et al. Synthesis, anti-
tumor and antimicrobial activity of some new 6-methyl-3-
phenyl-4(3H)-quinazolinone analogues: in silico studies. J
Enzyme Inhib Med Chem 2016;31:721–35.
fold: molecular docking studies. Eur
2016;115:121–31.
J
Med Chem
4. Alafeefy AM, El-Azab AS, Mohamed MA, et al. Synthesis of
some new substituted iodoquinazoline derivatives and
their antimicrobial screening. J Saudi Chem Soc 2011;15:
319–25.
5. Alanazi AM, Alaa A-M, Al-Suwaidan IA, et al. Design, synthe-
sis and biological evaluation of some novel substituted qui-
nazolines as antitumor agents. Eur
2014;79:446–54.
J
Med Chem
6. Alanazi AM, Al-Suwaidan IA, Alaa A-M, et al. Design, synthe-
sis and biological evaluation of some novel substituted 2-
mercapto-3-phenethylquinazolines as antitumor agents. Med
Chem Res 2013;22:5566–77.
7. Al-Obaid A, Abdel-Hamide S, El-Kashef H, et al. Synthesis, in
vitro antitumor activity and molecular modeling study of cer-
tain 2-thieno-4 (3H)-quinazolinone analogs. Eur J Med Chem
2009;44:2379–91.
ꢀ
23. Barlesi F, Tchouhadjian C, Doddoli C, et al. Gefitinib (ZD1839,
R
V
Iressa ) in non-small-cell lung cancer: a review of clinical tri-
als from a daily practice perspective. Fundam Clin Pharmacol
2005;19:385–93.
24. Barker AJ, Gibson K, Grundy HW, et al. Studies leading to
the identification of ZD1839 (IressaTM): an orally active,
selective epidermal growth factor receptor tyrosine kinase
inhibitor targeted to the treatment of cancer. Bioorg Med
Chem Lett 2001;11:1911–14.
25. Ganjoo KN, Wakelee H. Review of erlotinib in the treatment
of advanced non-small cell lung cancer. Biologics
2007;1:335–46.
26. Ibrahim HS, Abou-Seri SM, Tanc M, et al. Isatin-pyrazole ben-
zenesulfonamide hybrids potently inhibit tumor-associated
carbonic anhydrase isoforms IX and XII. Eur J Med Chem
2015;103:583–93.
27. Motzer RJ, Michaelson MD, Redman BG, et al. Activity of
SU11248, a multitargeted inhibitor of vascular endothelial
growth factor receptor and platelet-derived growth factor
receptor, in patients with metastatic renal cell carcinoma.
J Clin Oncol 2006;24:16–24.
28. Prenen H, Cools J, Mentens N, et al. Efficacy of the kinase
inhibitor SU11248 against gastrointestinal stromal tumor
mutants refractory to imatinib mesylate. Clin Cancer Res
2006;12:2622–7.
29. Teng Y-O, Zhao H-Y, Wang J, et al. Synthesis and anti-cancer
activity evaluation of 5-(2-carboxyethenyl)-isatin derivatives.
Eur J Med Chem 2016;112:145–56.
8. Al-Omar M, El-Azab A, El-Obeid S, Hamide HA. Sythesis of
some new 4-(3H)-quinazoline analogs as potential antioxi-
dant agents. Saudi J Chem Soc 2006;10:113–28.
9. Al-Omary FA, Abou-Zeid LA, Nagi MN, et al. Non-classical
antifolates. Part 2: synthesis, biological evaluation, and
molecular modeling study of some new 2, 6-substituted-qui-
nazolin-4-ones. Bioorg Med Chem 2010;18:2849–63.
10. Al-Suwaidan IA, Abdel-Aziz AA-M, Shawer TZ, et al.
Synthesis, antitumor activity and molecular docking study of
some novel 3-benzyl-4 (3H) quinazolinone analogues. J
Enzyme Inhib Med Chem 2016;31:78–89.
11. Al-Suwaidan IA, Alanazi AM, Alaa A-M, et al. Design, synthesis
and biological evaluation of 2-mercapto-3-phenethylquinazoline
bearing anilide fragments as potential antitumor agents:
molecular docking study. Bioorg Med Chem Lett 2013;23:
3935–41.
12. AzizaNassar M, AbdelHamide M, ElHakim S, El-Azab AA.
Synthesis and antimicrobial activities of some new 3-hetero-
aryl-quinazolin-4-ones. Indian J Heterocycl Chem 1996;6:
25–30.

944
A. S. EL-AZAB ET AL.
30. Koenig M, Cui J, Wei C, et al. Indolinone hydrazides as c-Met 33. Naglah AM, Shinwari Z, Bhat MA, et al. Targeting leukemic
inhibitors. Google Patents, 2006.
31. Al-Suwaidan IA, Alanazi AM, El-Azab AS, et al. Molecular
side population cells by isatin derivatives of nicotinic acid
amide. J Biol Regul Homeost Agents 2016;30:353.
design, synthesis and biological evaluation of cyclic 34. Chemical Computing Group. MOE: molecular operating
imides bearing benzenesulfonamide fragment as potential
COX-2 inhibitors. Part 2. Bioorg Med Chem Lett 2013;23:
2601–5.
environment. 2008.10 of Chemical Computing Group. Inc.
Available from: http://www.chemcomp.com/press_releases/
2008-11-04.htm
32. Radwan AA, Al-Mohanna F, Alanazi FK, et al. Target 35. Stamos J, Sliwkowski MX, Eigenbrot C. Structure of the epi-
b-catenin/CD44/Nanog axis in colon cancer cells by certain
dermal growth factor receptor kinase domain alone and in
complex with a 4-anilinoquinazoline inhibitor. J Biol Chem
2002;277:46265–72.
N0-(2-oxoindolin-3-ylidene)-2-(benzyloxy)
benzohydrazides.
Bioorg Med Chem Lett 2016;26:1664–70.