Antitumor and antileishmanial evaluation of novel heterocycles derived from quinazoline scaffold: A molecular modeling approach

Article abstract of DOI:10.1007/s00044-012-0213-9

Novel thiazole, pyrimidine and benzylidene derivatives derived from quinazoline scaffold have been synthesized. The antitumor evaluation of the newly synthesized products against three cancer cell lines namely breast adenocarcinoma (MCF-7), non-small cell

Full text of DOI:10.1007/s00044-012-0213-9

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res (2013) 22:2207–2221
DOI 10.1007/s00044-012-0213-9
ORIGINAL RESEARCH
Antitumor and antileishmanial evaluation of novel heterocycles
derived from quinazoline scaffold: a molecular modeling
approach
•
Daisy H. Fleita Rafat M. Mohareb
•
Ola K. Sakka
Received: 18 April 2012 / Accepted: 22 August 2012 / Published online: 9 September 2012
Ó Springer Science+Business Media, LLC 2012
Abstract Novel thiazole, pyrimidine and benzylidene
derivatives derived from quinazoline scaffold have been
synthesized. The antitumor evaluation of the newly synthe-
sized products against three cancer cell lines namely breast
adenocarcinoma (MCF-7), non-small cell lung cancer
(NCI-H460), and CNS cancer (SF-268) showed that the
benzylidene–quinazoline derivative 12a showed remarkable
activity against all three cell lines. The thiazolo-quinazoline
derivative 10 showed greater activity than the control against
breast adenocarcinoma (MCF-7) with a concentration of
0.01 lM. Moreover, the antileishmanial activity of the
newly synthesized products tested on Leishmania donovani
amastigotes showed that compounds 4, 14, and 18 had very
promising activity.
(Sordella et al., 2004; Zhang et al., 2007). Quinazolines are
the most promising small-molecule selective EGFR-TK
inhibitors (Fry et al., 1994; Rewcastle et al., 1995; Palmer
et al., 1997). Gefitnib and Erlotinib are examples of anili-
noquinazolines which have occupied a promising section in
the anticancer market (Fig. 1). Both drugs are EGFR tyro-
sine kinase inhibitors which act by binding to the adenosine
triphosphate (ATP) site of the enzyme, thus inhibiting
malignant cells. They are currently marketed as IressaÓ and
TarcevaÓ and were recently approved for the treatment of
HER2-positive metastatic breast cancer (Hennequin et al.,
2002; Wood et al., 2004; Burris et al., 2005). PTKs are also
key enzymes of mammalian signal transduction, thus their
identification and mechanism characterization are essential
to elucidate host–parasite interactions and parasite biology.
Recent studies have shown that PTKs are involved in the
uptake of virulent and avirulent Leishmania donovani
promastigotes by macrophage cells (Gosh and Chakraborty,
2002). The search for new PTK inhibitors with observable
structural diversity has been an active aspect of current
pharmaceutical research endeavors involving both cancer
and leishmaniasis (El-Azab et al., 2010). Quinazolinone
derivatives have also been found to possess a potent wide
spectrum of activities like antibacterial (Hassanzadeh et al.,
2012), antifungal (Xu et al., 2011), anticonvulsant (Kornet
et al., 1983), and anti-inflammatory (Ozaki et al., 1985;
Daidone et al., 1994). In addition, several quinazolines were
also demonstrated to be potent antileishmanials (Gilbert,
2002; Khabnadideh et al., 2005). Most of the SAR resear-
ches focused on the variation of substitutions at the 6- and
7-positions as well as substitutions at the 4-anilino portion
of 4-anilino quinazolines. To our knowledge, limited
research has been done on the variation of substitutions at
the 2 and 3 positions of the quinazoline nucleus (Wright
et al., 2001; Toledo-Sherman et al., 2005). In the light of
Keywords Quinazolines ꢀ Thiazole ꢀ Pyrimidine ꢀ
Antitumor ꢀ Antileishmanial
Introduction
Protein tyrosine kinases (PTKs) are known for their role in
cancer. The epidermal growth factor receptor (EGFR) is a
member of the PTKs’ receptors family. Overexpression of
EGFR leading to uncontrolled cell proliferation has been
proven to be a definite cause of a significant number of
human tumors (e.g., breast, ovarian, colon, and prostate)
D. H. Fleita ꢀ O. K. Sakka
Department of Chemistry, American University in Cairo,
P.O. Box 74, New Cairo 11835, Arab Republic of Egypt
R. M. Mohareb (&)
Department of Chemistry, Faculty of Science, Cairo University,
Giza, Arab Republic of Egypt
e-mail: raafat_mohareb@yahoo.com
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Med Chem Res (2013) 22:2207–2221
Fig. 1 Examples of EGFR–TK
inhibitors
such observations, our efforts toward drug discovery
prompted us to design, synthesize, and evaluate the biologic
activity of some new quinazoline derivatives as antitumor
and antileishmanial agents. A new series of quinazolinone
derivatives were synthesized by incorporation of a variety
of ring systems such as pyrimidines, thiazoles, coumarins,
pyrazoles, and benzylidenes at the 3rd position of the qui-
nazoline pharmacophore, the introduction of a methyl group
at the 2nd position, and a carbonyl group at position 4.
ethyl 2-(2-methyl-4-oxoquinazolin-3(4H)-yl-amino) acetate 2
via the loss of HCl. On the other hand, carrying the same
reaction in an oil bath at 120 °C afforded 2-chloro-N-(2-
methyl-4-oxoquinazolin-3(4H)-yl)acetamide 3 via the loss of
ethanol. The structures of compounds 2 and 3 were established
on the basis of analytical and spectral data. Thus, the 1H NMR
spectrum of compound 2 showed the presence of a singlet at d
2.48 indicating the presence of the CH3 group, a triplet at d 2.58
equivalent to the ester CH3, a singlet (D2O exchangeable) at d
2.60 for the NH, a quartet at d 3.32 indicating the ester CH2, a
singlet at d 5.79 for the a-carbonyl-CH2 group, and a multiplet
at d7.44–8.11 corresponding to the phenyl protons. Compound
1 reacted readily with phenyl isothiocyanate to give the
phenylthiourea derivative 4. The analytical and spectral data of
the latter product is analogous with its assigned structure (see
Experimental section).
Results and discussion
In an attempt to reach an active antitumor agent with enhanced
activity and selectivity, the substitution pattern at the 2nd and
3rd positions of the quinazoline pharmacophore were selected
in order to alter the electronic environment and thus affect the
lipophilicity and hence the activity of the target molecules. Our
aim was the removal of bulky fragments at the 4th and 6th
position of the quinazoline nucleus and the introduction at the
3rd position of a variety of aryl systems such as pyrimidines
(Capdeville et al., 2002), thiazoles (El-Subbagh et al., 1994;
Schnur et al., 1991), coumarins (Madari et al., 2003; Kostova,
2005), and pyrazoles (Xia et al., 2007; Farag et al., 2008) that
are known to contribute to the enhancement of the antitumor
activity. The incorporation of a methyl group at the 2nd posi-
tion of the quinazoline pharmacophore was of great importance
in order to increase the lipophilicity and thus drastically modify
the bioavailability and efficacy of our synthesized series
(Fig. 2). Our synthetic methodology employed the use of ethyl
2-(2-methyl-4-oxoquinazolin-3(4H)-ylamino) acetate 2 as our
first building block, 2-chloro-N-(2-methyl-4-oxoquinazolin-
The reaction of compound 2 with either hydrazine
hydrate or phenylhydrazine gave the acetohydrazide
derivatives 5a or 5b, respectively, via ethanol elimination.
On the other hand, compound 2 reacted with salicylalde-
hyde in the presence of pyridine to form the coumarin
derivative 6. Moreover, compound 2 reacted with o-phen-
ylenediamine to give the 3-imidazol-2-ylquinolone deriv-
ative 7. The structure of the latter product was confirmed
on the basis of analytical and spectral data. Thus, its 13C
NMR spectrum showed the presence of d: 20.1 (quinaz-
olinone–CH3), 115.3, 120.9, 122.4, 123.0, 127.4, 128.8,
133.5, 138.9, 141.1, and 147.1 (benzene, imidazole), 161.1
(C=O, quinazolinone), 164.4 (C–CH3–quinazolinone).
Moreover, compound 2 reacted with either malononitrile or
ethyl cyanoacetate in the presence of triethylamine to give
the pyrrole derivatives 9a or 9b, respectively (Scheme 2).
Next, the reaction of a-haloketone 3 with thiourea was
studied as an application of Hantzsch thiazole synthesis,
and gave the thiazole derivative 10. Moreover, the reaction
of 3 with salicylaldehyde gave the benzofuran derivative
11, while its reaction with benzaldehyde, 4-chlorobenzal-
dehyde, or 4-methoxybenzaldehyde gave the correspond-
ing benzylidene derivatives 12a–c. The analytical and
spectral data of compounds 9a–b, 10, 11, and 12 a–c are in
agreement with their respective structures.
3(4H)-yl)acetamide
3 as our second building block,
1-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-phenylthiourea 4 as
our third building block, and 3-amino-4(3H)-quinazolinone 1
as our scaffold. The reaction sequence is illustrated in
Scheme 1. The reaction of 3-amino-4(3H)-quinazolinone 1
withethylchloroacetate wascarriedoutby twodifferentroutes.
The starting material 1 was prepared according to a reported
procedure (Alagarsamy et al., 2005). The reaction of com-
pound 1 and ethyl chloroacetate in refluxing ethanol afforded
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Med Chem Res (2013) 22:2207–2221
2209
Fig. 2 Proposed fragment
replacement to obtain
synthesized quinazoline
derivatives
Next, we shifted our efforts toward studying the reac-
tivity of 3 toward different chemical reagents. Thus, mal-
ononitrile reacted with phenylisothiocyanate in DMF/KOH
solution at room temperature to give the corresponding
intermediate potassium sulfide salt which reacted readily
with the chloromethylene moiety present in compound 3 to
give the thiazole derivative 13. The analytical and spectral
data were the basis of its structure elucidation (see
Experimental section).
a-bromoacetonitrile in ethanol under reflux gave the thia-
zole derivatives 17 or 18, respectively (Scheme 4).
Effect of the synthesized compounds on the growth
of human tumor cell lines
The synthesized compounds (2–18, Schemes 1, 2, 3, 4)
were subjected to the NCI’s in vitro, one dose primary
anticancer assay, using a 3-cell line panel consisting of
MCF-7 (breast), NCI-H460 (lung), and SF-268 (CNS)
cancers. The results are summarized in Table 1. The data of
compounds 2–18 in Table 1 summarize the effect of dif-
ferent ring systems on their activity. With regard to sensi-
tivity against individual cell lines, compound 9a showed
considerable activity against MCF-7 (breast adenocarci-
noma), NCI-H460 (non-small cell lung cancer), and SF-268
(CNS cancer) with GI50 concentrations of 0.4, 0.1, and
0.9 lM, respectively. Compound 10 showed the highest
potency against MCF-7 (breast adenocarcinoma) at a con-
centration of 0.01 lM, and is thus almost twofolds more
active than the positive control doxorubicin. Compound 10
also exhibited moderate activity against NCI-H460 (lung)
Compound 3 reacted with o-aminobenzoic acid and
afforded the indolin-3-one derivative 14 (Scheme 3). The
analytical and spectral data of the latter compound are
consistent with the assigned structure. On the other hand,
the reaction of 4 with ethyl acetoacetate in 1,4-dioxane
solution containing a catalytic amount of piperidine
afforded the pyrimidine derivative 15. Formation of the
latter product was explained in terms of ethanol and water
elimination. Similarly, the reaction of 4 with acetylacetone
afforded the pyrimidine derivative 16 via two molecules of
1
water elimination. The H NMR and 13C NMR spectra of
compounds 15 and 16 were the basis of their structure
elucidation. On the other hand, the reaction of compound 4
with its thioamide moiety with either ethyl chloroacetate or
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S
O
O
H
H
N
N
CH2COOC2H5
C
NHPh
N
N
CH3
N
N
CH3
2
4
PhNCS
H2C COOC2H5
Cl
EtOH 24 h
O
EtOH/Et3N
heat 2h
NH2
N
CH3
N
1
H2C COOC2H5
Cl
oil bath, 120 oC
O
H
O
N
C CH2Cl
N
CH3
N
3
Scheme 1 Synthesis of compounds 2, 3, and 4
O
O
N
H2
H
O
H
N
H
N
C
C NHNHR
N CH2
N
N
5a, R=H
N
5b, R= Ph
CH3
N
CH3
RNHNH2
NH2
R = H
7
R = Ph
EtOH/heat
2h
NH2
O
EtOH/heat
3h
H
N
CH2COOC2H5
N
CH3
H2C
CN
X
N
CHO
OH
2
8a, X=CN
8b, X=COOEt
EtOH/Et3N
heat 3h
O
O
Y
1,4-dioxane/piperidine
O
H
N
NH2
N
O
N
N
CH3
N
COOEt
N
CH3
6
9a, Y=NH2
9b, Y=OH
Scheme 2 Synthesis of compounds 5a, b, 6, 7, and 9a, b
and SF-268 (CNS) cancers at a concentration of 0.2 and
0.4 lM, respectively. Compound 12a showed a remarkable
activity against MCF-7 (breast adenocarcinoma),
NCI-H460 (non-small cell lung cancer), and SF-268 (CNS
cancer) with GI50 concentrations of 0.03, 0.02, and
0.05 lM, respectively. However, compound 12b showed
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Med Chem Res (2013) 22:2207–2221
2211
Scheme 3 Synthesis of
compounds 10, 11, 12a–c,
and 14
O
C
O
N
H
N
C
Cl
O
N
H
N
N
N
NH2
N
Ar
CH3
S
CH3
12
a
S
Ar
ArCHO
10
C6H5
H2N
C
NH2
1,4-dioxane/piperidine
EtOH, heat 3h
b
4-OCH3-C6H4
4-Cl-C6H4
c
O
O
H
N
C
CH2Cl
CHO
OH
N
NH2
CH3
N
1,4-dioxane/piperidine
3
COOH
EtOH, heat 1.5 h
O
NC
O
CN
H
N
S-K+
C
O
N
Ph N
H
HN
C
N
CH3
O
N
H
N
CN
11
NC
N
N
O
O
CH3
Ph
S
14
O
N
N
N
H
13
CH3
O
H3C
N
O
O
N
N
C6H5
C6H5
N
O
S
N
S
CH3
N
N
O
O
15
N
CH3
O
H3C
OEt
Cl
17
1,4-dioxane/piperidine
OEt
heat 3h
EtOH/ heat 3 h
S
O
H
N
C
NHC6H5
N
O
O
N
CH3
H2C Br
CN
EtOH/heat 3h
H3C
CH3
4
1,4-dioxane/piperidine
heat 3h
NH2
CH3
H3C
C6H5
N
O
N
N
O
N
C6H5
S
N
N
S
N
N
CH3
CH3
16
18
Scheme 4 Synthesis of compounds 15, 16, 17, and 18
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Med Chem Res (2013) 22:2207–2221
Table 1 Effect of compounds 2–18 on the growth of three human tumor cell lines
Compound no.
GI50 (lM)
Compound no.
GI50 (lM)
MCF-7
NCI-H460
22.6 8.9
SF-268
MCF-7
NCI-H460
12.1 0.5
SF-268
O
H
20.9 0.6
50.1 0.7
26.8 0.5
10.9 0.8
22.3 0.5
O
O
N CH2COOC2H5
H
N
N
N
C
O
CH3
N
2
3
N
O
CH3
11
O
C
O
C
O
N
23.2 4.8
18.4 1.8
0.03 0.001 0.02 0.003 0.05 0.01
H
N
H
N
C
Cl
CH2Cl
N
N
CH3
N
O
CH3
12a
S
O
O
64.9 12.8
11.8 0.6
78.6 11.6
14.5 0.8
56.8 5.9
16.7 1.6
0.9 0.09
0.6 .08
0.8 0.06
H
H
N
C
C
Cl
N
C
NHPh
N
N
CH3
N
O
N
CH3
OCH3
4
12b
12c
O
C
70.6 16.9
38.9 10.8
50.8 8.6
O
H
N
O
N
O
N
H2
C
H
N
C
Cl
C
NHNH2
N
N
N
N
CH3
CH3
5a
Cl
O
C
CN
S
H2
C
NC
N
22.6 0.4
12.9 0.1
24.8 1.9
40.6 12.2
32.6 8.6
60.4 14.8
H
N
NHNH
Ph
O
N
CH3
N
N
H
5b
6
CH3
13
14
O
O
32.0 0.8
17.3 1.4
22.3 1.5
11.9 0.6
14.1 0.6
20.3 0.5
H
N
HN
O
N
O
N
H
N
C
N
CH3
N
O
O
O
CH3
O
N
H3C
N
20.0 0.6
22.0 0.4
31.5 8.0
22.9 0.8
43.6 1.8
36.8 23.5
H
H
N
CH2
O
N
N
N
N
N
C6H5
S
CH3
N
CH3
7
15
16
CH3
H3C
H2N
0.4 0.05
13.1 0.07
0.01 0.002
0.1 0.04
0.9 0.03
0.2 0.3
0.9 0.02
11.5 0.03
0.4 0.06
0.7 0.05
22.0 0.2
36.0 1.6
0.2 0.08
30.6 1.4
28.0 3.9
0.03 0.01
38.4 0.6
22.5 2.4
O
N
N
N
NH2
COOEt
O
C6H5
N
N
S
N
CH3
CH3
N
O
9a
9b
O
HO
C6H5
N
O
N
NH2
N
N
N
N
S
CH3
N
CH3
COOEt
17
18
O
N
NH2
H
N
N
C6H5
NH2
N
N
O
N
S
CH3
S
N
N
10
CH3
Doxorubicin
Doxorubicin
0.04 0.008 0.09 0.008 0.09 0.007
Results are given in concentrations that were able to cause 50 % of cell growth inhibition (GI50) after continuous exposure of 48 h and show
mean SEM of three independent experiments performed in duplicate
less potency against the three cell lines with GI50 concen-
trations of 0.9, 0.6, and 0.8. Compound 16 showed the best
inhibitory effect against SF-268 (CNS cancer) with a GI50
concentration of 0.03 lM. Compounds 4 and 12c showed
the least inhibitory effect against the three cell lines.
Structure activity correlation
Structure–activity correlation of the synthesized com-
pounds revealed that the substitution of unsubstituted
benzylidenes into the 3rd position of the quinazoline
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Med Chem Res (2013) 22:2207–2221
2213
pharmacophore greatly enhances the cytotoxicity of the
compound. Upon inspecting the benzylidene–quinazoline
series 12a–c, it is obvious that in 12a, GI50 values of 0.03,
0.02, and 0.05 lM, the unsubstituted benzene ring moiety
favored the antitumor activity of all cell lines. The addition
of an electron-donating methoxy group slightly decreased
the GI50 concentrations of the three cell lines to 0.9, 0.6,
and 0.8 lM, respectively. On the other hand, the addition
of an electron-withdrawing chlorine group markedly
decreased the activity in 12c. Substitution of a pyrimidine
ring into the 3rd position of the quinazoline ring yielded
compound 16 (GI50 value 0.03 lM) as the most active
member of this study against SF-268 (CNS cancer).
However, the incorporation of an oxo-pyrimidine func-
tionality in 15 did not result in a similar enhancement of
antitumor activity. Replacement of the pyrimidine group
with the thiazole group and the introduction of a spacer
amino function yielded compound 10 (GI50 value
0.01 lM), which is almost twofolds more active against
MCF-7 than the positive control doxorubicin used in our
study. The addition of a bulky phenyl group to the thiazole
ring in derivative 18 resulted in a decrease in the inhibitory
effect. A similar effect was observed in derivative 13.
Similarly, the incorporation of an oxo-thiazole functional-
ity in 17 did not result in a similar enhancement of anti-
tumor activity as compared to the effect brought about by
the thiazole ring. Derivatives 6 and 7 carrying the coumarin
and imidazole functionality did not show a remarkable
enhancement in antitumor activity. Similarly, derivatives 2,
3, and 5a which lacked the aryl functionality showed no
considerable enhancement in cytotoxicity. Pyrazolo-quin-
azolines 9a and 9b showed moderate cytotoxicity; how-
ever, 9a, the pyrazole ring of which includes an amino
function, showed better cytotoxicity than 9b containing a
hydroxyl-function.
Table 2 Antileishmanial activity of compounds 2–18 at 50 lM
against L. donovani axenic amastigotes
Compound Average
no.
GI50
Compound Average
GI50
inhibition (lM)
(%)
inhibition (lM) no.
(%)
2
44
60
98
58
22
70
85
40
60
72
98
10
–
11
28
80
84
50
44
92
66
48
44
96
0
14
26
28
18
18
13
12
–
3
12a
12b
12c
13
4
12
22
28
–
5a
5b
6
14
7
–
15
9a
9b
10
–
16
18
22
17
12
10
18
Positive
controla
Negative
controlb
GI50 = concentration for 50 % growth inhibition
a
Amphotericin B (1 lM)
c
Culture medium and DMSO
resulted in a remarkable increase in the average inhibition.
Comparing benzylidene–quinazoline derivatives 12 a–c, it
is of great value to note that chloro benzylidene derivative
12b exhibited the highest inhibition as compared to its
counter analogs 12a and 12c.
Druglikeness
In recent years, one of the tools for predicting druglikeness,
which discriminates between drug-like and non drug-like
compounds, is the Lipinski’s rule of five (Lipinski et al.,
2001), which takes into consideration molecular weight,
hydrophilicity (cLogP), number of hydrogen bond donors,
and number of hydrogen bond acceptors. According to the
results obtained by the Molinspiration software, none of the
most antitumor and antileishmanial active compounds
(compounds 4, 18, 16, 10, 12a, and 14) violate any of the
Lipinski’s criteria, an important characteristic for future
drug development. Additional SAR parameters were cal-
culated by means of the Osiris program such as solubility
(LogS), druglikeness, and drug score. Osiris program
druglikeness values are calculated from 15,000 Fluka
compounds and from 3,300 traded drugs. A positive value
states that your molecule contains predominantly frag-
ments which are frequently present in commercial drugs.
The drug score combines druglikeness, cLogP, logS,
molecular weight in one value that may be used to judge
the compound’s overall potential to qualify for a drug. A
high solubility and a low hydrophilicity contribute to a
compound’s absorption or permeation ability. Compound
Antileishmanial activity
Results indicated that among the 20 synthesized quinazo-
line derivatives analyzed, phenyl thiourea derivative 4,
thiazole derivative 18, and indole derivative 14 displayed
the most significant activity against L. donovani amastig-
otes growing in macrophages similar to that seen with
axenic amastigotes at 50 lM with an average inhibition of
98, 96, and 92 %, respectively. The results are summarized
in Table 2. It is obvious that 1-(2-methyl-4-oxoquinazolin-
3(4H)-yl)-3-phenylthiourea 4 showed maximum average
inhibition similar to the positive control Amphotericin B
used in our study. On the other hand, compounds 5b and 11
showed the lowest activity. Upon inspection of thiazole
analogs 18 and 10, it can be inferred that the addition of a
bulky phenyl group to the thiazole ring in derivative 18
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Med Chem Res (2013) 22:2207–2221
14 which displayed high antileishmanial activity possessed
the lowest hydrophilicity, a reasonably high solubility, and
a high drug score compared to all active compounds. On
the other hand, compound 10 which displayed the highest
activity against breast cancer possessed the highest solu-
bility and the highest drug score among all active com-
pounds. Thus, there is a good correlation between
calculated SAR parameters and the biologic activity of our
most active compounds (Table 3).
Docking study
Structural models
To compare the binding mode of most active compounds
10 (0.01 lM; MCF-7) and 12a (0.03 lM; MCF-7) to the
known EGFR inhibitor Erlotinib, a docking study into the
ATP binding site of EGFR was performed. The ATP
binding site of EGFR has the following features. Adenine
region: contains two key hydrogen bonds formed by the
interaction of the purine base of ATP and the protein
backbone between amino acids Gln767 and Met769.
Hydrophobic pockets: Though not used by ATP, plays an
important role in inhibitor selectivity. Phosphate binding
region: This is used for improving inhibitor selectivity
(Fabbro et al., 2002) (Fig. 3a). Erlotinib binds to the ade-
nine region of EGFR by mimicking the binding of ATP
with the protein backbone via an interaction between the
N-1 of the quinazoline which accepts an H-bond from the
backbone Met-769 amide nitrogen existing in the hinge
region of EGFR. Another significant interaction comes
from the presence of a water-mediated hydrogen bond
between N-3 of quinazoline ring and a water molecule near
Thr-766 side chain of EGFR molecule. Our preliminary
study of erlotinib self-docking to EGFR-TK was in
agreement with the reported literature (Stamos et al.,
2002). A binding energy of -10.53 kcal/mol was obtained
Fig. 3 a Erlotinib (green) and compound 10 (purple) docked into the
binding site of EGFR. b Erlotinib (green) and compound 12a (purple)
docked into the binding site of EGFR. Hydrogen bonding interactions
are shown in red
for compound 10. Our docking studies on compound 10
have revealed that N-1 of the quinazoline forms a hydrogen
bonding interaction with basic Lys-721 side chain residue
Table 3 Physico-chemical and absorption properties for the most active compounds
Compound
cLogP
MW
n-OHNH donorsa
n-ON acceptorsb
Lipinski’s violations
Log Sc
Druglikeness
Drug score
4
2.07
2.38
2.9
310
349
376
273
339
334
2
1
0
2
1
2
5
6
5
0
5
7
0
0
0
0
0
0
-3.93
-6.09
-5.81
-3.09
-4.39
-3.76
2.3
0.77
0.56
0.34
0.87
0.71
0.76
18
16
10
12a
14
5.68
3.35
4.45
2.64
6.08
2.36
2.68
0.65
cLogP logarithm of compound partition coefficient between n-octanol and water; MW molecular weight
a
n-OHNH number of hydrogen bond donors
b
n-ON number of hydrogen bond acceptors
c
Aqueous solubility
123

Med Chem Res (2013) 22:2207–2221
2215
NCI-H460, and SF-268 with GI50 range of 0.01, 0.02, and
0.05 lM, respectively. Since overexpression of EGFR is
related to breast cancer, molecular docking studies of
EGFR were critical for understanding the antitumor
selectivity of compound 10. Our molecular docking studies
for compound 10 revealed the interactions between our
prepared compound and the binding site of EGFR, and it
could be postulated that compound 10 might act on the
same enzyme target where Erlotinib acted. On the other
hand, results indicated that compound 4 displayed the most
significant activity against L. donovani amastigotes.
Calculation of SAR parameters complemented our exper-
imental results. Thus, this work paves the way to discov-
ering promising leads for antitumor and antileishmanial
drugs in the future.
Fig. 4 3D visualization of Erlotinib in the binding site of EGFR
˚
which exists in the hydrophobic region (2.5 A). In addition,
Experimental section
the NH2 group located on the thiazole ring donates a
hydrogen to Leu 764 residue present in the backbone of
˚
EGFR (2.9 A). A final interaction that is analogous to
Docking study
erlotinib is the presence of a water-mediated hydrogen
bond between NH group directly attached to quinazoline
ring and a water molecule near Thr-766 side chain of
Structure coordinates for EGFR were obtained from RCSB
Protein Data Bank (PDB ID: 1M17). The 3D structure of
compounds 10 and 12 were obtained by means of Chem-
draw Ultra. The docking study of the designed compounds
into EGFR tyrosine kinase was performed by means of
‘‘Molecular Operating Environment (MOE) version
2008.10 release of Chemical Computing Groups.’’ The
complexes were energy-minimized with a MMFF94 force-
field till the gradient convergence 0.01 kcal/mol was
reached (Halgren, 1996).
˚
EGFR (2.9 A) (Fig. 3a). Compound 10 docked into the 3D
surface of EGFR is shown in Fig. 4. Interestingly, docking
of compound 12 into the active site of EGFR indicated only
the presence of a water-mediated hydrogen bond between
the oxygen atom of the carbonyl group and a water mol-
˚
ecule near Thr-766 side chain (2.7 A) (Fig. 3b). Compar-
ing the docking modes of compounds 10 and 12a, it could
be postulated that compound 10 might bind even better
than compound 12a to EGFR since it forms three hydrogen
bonding interactions with the ATP binding site, while the
latter forms only one interaction. This comes in agreement
with the experimental values since compound 10 was the
most potent against MCF-7 with a concentration of
0.01 lM. However, 10, 12a, and erlotinib share a common
water-mediated hydrogen bond near Thr-766 residue which
indicates that it might be a structural necessity for their
activity.
Antitumor evaluation
Materials, methods, and reagents
Fetal bovine serum (FBS) and L-glutamine were from
Gibco Invitrogen Co. (Scotland, UK). RPMI-1640 medium
was from Cambrex (New Jersey, USA). Dimethyl sulfox-
ide (DMSO), doxorubicin, penicillin, streptomycin, and
sulforhodamine B (SRB) were from Sigma Chemical Co.
(Saint Louis, USA). Samples: Stock solutions of com-
pounds 2–18 were prepared in DMSO and kept at 20 °C.
Appropriate dilutions of the compounds were freshly pre-
pared just prior to the assays. Final concentrations of
DMSO did not interfere with the cell growth.
Conclusion
The present work led to the development of novel
antitumor and antileishmanial molecules, containing the
3-substituted quinazolinone pharmacophore. Three human
tumor cell lines including MCF-7 (breast adenocarcinoma),
NCI-H460 (non-small cell lung cancer), and SF-268 (CNS
cancer) were used to measure the cytotoxic activity of the
proposed quinazolinone derivatives. Compounds 10 and
12a showed the highest inhibitory effect against MCF-7,
Cell cultures
Three human tumor cell lines MCF-7 (breast adenocarci-
noma), NCI-H460 (non-small cell lung cancer), and
SF-268 (CNS cancer) were used. MCF-7 was obtained
from the European Collection of Cell cultures (ECACC,
123

2216
Med Chem Res (2013) 22:2207–2221
Salisbury, UK) and NCI-H460 and SF-268 were kindly
provided by the National Cancer Institute (NCI, Cairo,
Egypt). They grow as monolayer and are routinely main-
tained in RPMI-1640 medium supplemented with 5 % heat
inactivated FBS, 2 mM glutamine, and antibiotics (peni-
cillin 100 U/mL, streptomycin 100 lg/mL) at 37 °C in a
humidified atmosphere containing 5 % CO2. Exponentially
growing cells were obtained by plating 1.5 9 105 cells/mL
for MCF-7 and SF-268 and 0.75 9 104 cells/mL for NCI-
H460, followed by 24 h of incubation. The end point
determinations were made with a protein binding dye,
SRB. The effect of the vehicle solvent (DMSO) on the
growth of these cell lines was evaluated in all the experi-
ments by exposing untreated control cells to the maximum
concentration.
m.p. 135 °C; IR (KBr) t/cm-1: 3,577 (NH), 3,067 (CH-
aromatic), 2,930 (CH3), 2,867 (CH2),1,680 (C=N), 1,675
1
(C=O); MS m/z (%) 261 [M?, 13.58 %]; H-NMR d: 2.48
(s, 3H, quinazolinone–CH3), 2.58 (t, 3H, J = 7.22 Hz,
CH3), 2.60 (s, 1H, NH), 3.32 (q, 2H, J = 7.22 Hz CH2),
5.79 (2H, s, CH2C=O), 7.44–8.11 (4H, m, Ar–H);
13C-NMR d: 14.1 (CH3), 20.2 (quinazolinone–CH3), 52.1
(CH2–C=O),61 (CH2), 120.9, 122.4, 127.4, 128.8, 133.5,
147.1 (benzene),161.1 (C=O), 164.2 (C–CH3–quinazoli-
none), 169.6 (C=O); Anal. Cald. for C13H15N3O3 (261.28):
C, 59.76; H, 5.79; N, 16.08 %. Found: C, 59.70; H, 5.69;
N, 16.11 %.
2-Chloro-N-(2-methyl-4-oxoquinazolin-3(4H)-yl)
acetamide (3)
General
Equimolecular amounts of compound 1 (1.75 g, 0.01 mol)
and ethyl chloroacetate (1.22 g, 0.01 mol) were heated in
an oil bath at 120 °C for 30 min. After cooling, the reaction
mixture was heated in ethanol, then poured onto an ice/
water mixture, and the formed solid product was collected
by filtration and crystallized from ethanol. Yield: 1.63 g,
65 %, m.p. 125 °C; IR (KBr) t/cm-1: 3,580 (NH), 3,070
(CH-aromatic), 2,910 (CH3), 2,867 (CH2),1,680 (C=N),
Melting points were determined on an electrothermal
melting point apparatus (Electrothermal 9100) and are
uncorrected. IR spectra were recorded for KBr disks on a
1
Pye Unicam SP-1000 spectrophotometer. H- NMR, and
13C-NMR spectra were measured on a Varian EM-390-
200 MHz in CD3SOCD3 as solvent using TMS as the
internal standard, and chemical shifts are expressed as d.
Analytical data were obtained from the Microanalytical
Data Unit at Cairo University, Giza, Egypt. Antitumor
evaluation for the newly synthesized products was per-
formed by a research group at the National research center
and the National Cancer Institute at Cairo University. FBS
and L-glutamine were from Gibco Invitrogen Co.
(Scotland, UK). RPMI-1640 medium was from Cambrex
(New Jersey, USA). DMSO, doxorubicin, penicillin, strep-
tomycin, and SRB were from Sigma Chemical Co. (Saint
Louis, USA). Samples: Stock solutions of all compounds
were prepared in DMSO and kept at -20 °C. Appropriate
dilutions of the compounds were freshly prepared just prior
to the assays. Final concentrations of DMSO did not interfere
with the cell growth. MCF-7 was obtained from the Euro-
pean Collection of Cell Cultures (ECACC, Salisbury, UK)
and NCI-H460 and SF-268 were kindly provided by the
National Cancer Institute (NCI, Cairo, Egypt).
1
1,675 (C=O); MS m/z (%) 251 [M?, 1.34 %]; H-NMR d:
3.31 (s, 3H, quinazolinone–CH3), 4.09 (s, 2H, CH2Cl),
7.06–7.60 (m, 4H, Ar–H), 11.01 (s, 1H, NH); 13C-NMR d:
19.9 (quinazolinone–CH3), 44.9 (CH2–Cl), 122.4, 122.5,
127.4, 128.8, 133.5, 147.1 (benzene),161.4 (C=O), 164.1
(C–CH3–quinazolinone), 166.2 (C=O); Anal. Cald. for
C11H10ClN3O2 (251.67): C, 52.5; H, 4.01; N, 16.70 %.
Found: C, 52.3; H, 4.07; N, 16.75 %.
1-(2-Methyl-4-oxoquinazolin-3(4H)-yl)-3-phenylthiourea
(4)
Equimolecular amounts of compound 1 (1.75 g, 0.01 mol)
in ethanol (20 mL) and phenylisothiocyanate (1.35 g,
0.01 mol) were stirred for 24 h. The reaction mixture was
then poured onto an ice/water mixture and the formed solid
product was collected by filtration and crystallized from
ethanol. Yield: 1.96 g, 63 %, m.p. 180 °C; IR (KBr)
t/cm-1: 3,577–3,370 (2 NH), 3,110 (CH-aromatic), 2,930
(CH3), 1,675 (C=N), 1,675 (C=O), 1,241 (C=S); MS m/z
Ethyl 2-(2-methyl-4-oxoquinazolin-3(4H)-ylamino)acetate
(2)
1
(%) 310 [M?, 11.22 %]; H-NMR d: 2.58 (3H, s, quinaz-
olinone–CH3), 6.52–7.63 (m, 9H, Ar–H), 4.70 (s, 1H, –N–
NH–CS), 9.70 (s, H, CSNHPh); 13C-NMR d: 20.5 (qui-
nazolinone–CH3), 120.9, 122.4, 124.8, 126.5, 127.4, 128.8,
129.1, 133.5, 137.1, 147.1 (benzene), 161.4 (C=O), 164.3
(C–CH3–quinazolinone), 181.5 (C=S); Anal. Cald. for
C16H14N4OS (310.37): C, 61.92; H, 4.55; N, 18.05; S,
10.33 %. Found: C, 62.1; H, 4.49; N, 18.13; S, 10.29 %.
To a solution of compound 1 (1.75 g, 0.01 mol) in ethanol
(50 mL) containing triethylamine (0.5 mL), ethyl chloro-
acetate (1.22 g, 0.01 mol) was added. The reaction mixture
was heated under reflux for 2 h, and then poured onto ice/
water. The formed solid product was collected by filtration,
dried, and crystallized from ethanol. Yield: 1.75 g, 67 %,
123

Med Chem Res (2013) 22:2207–2221
2217
General procedure for the synthesis of 5a or 5b
quinazolinone–CH3), 3.36 (s, 1H, NH–CH2), 6.15–8.15 (m,
8H, Ar–H), 7.00 (s, 1H, coumarin H-4), 13C-NMR d: 20.3
(quinazolinone–CH3),113.5, 120.9, 121.5, 122.3, 122.4,
125.5, 126.8, 127.4, 128.4, 128.8, 133.5, 137.9, 147.1,
150.2 (benzene, coumarin), 159.9 (C=O, coumarin),161.1
(C=O, quinazolinone),164.5 (C–CH3–quinazolinone);
Anal. Cald. for C18H13N3O3 (319.31): C, 67.71; H, 4.10; N,
13.16 %. Found: C, 67.89; H, 4.23; N, 13.19 %.
To a solution of compound 2 (2.61 g, 0.01 mol) in ethanol
(30 mL), either hydrazine hydrate (0.5 g, 0.01 mol) or
phenyl hydrazine (1.18 g, 0.01 mol) was added. The
reaction mixture was heated under reflux for 2 h, and the
excess ethanol was distilled off. The formed solid product
in each case was collected by filtration, dried, and crys-
tallized from ethanol.
3-(1H-Benzo[d]imidazol-2-ylamino)-2-methylquinazolin-
4(3H)-one (7)
2-(2-Methyl-4-oxoquinazolin-3(4H)-ylamino)
acetohydrazide (5a)
To a solution of compound 2 (2.61 g, 0.01 mol) in ethanol
(30 mL), o-amino aniline (1.08 g, 0.01 mol) was added.
The reaction mixture was heated under reflux for 3 h, and
then excess solvent was distilled off. The formed solid
product was collected by filtration, dried and crystallized
from ethanol. Yield: 1.45 g, 66 %, m.p. 122 °C; IR (KBr)
t/cm-1: 3,330 (NH), 3,090 (CH-aromatic), 2,895 (CH3),
1,680 (C=N), 1,675 (C=O); MS m/z (%) 305 [M?,
Yield: 1.65 g, 67 %, m.p. 130 °C; IR (KBr) t/cm-1
3,459–3,330 (NH2, NH), 3,090 (CH-aromatic), 2,895
:
(CH3), 2,870 (CH2), 1,680 (C=N), 1,675 (C=O); MS m/z
1
(%) 247 [M?, 11.22 %]; H-NMR d: 2.58 (s, 3H, quinaz-
olinone CH3), 2.60 (s, 1H, NH–NH2), 3.29 (s, 2H, NH2),
3.54 (s, 2H, CH2), 7.44–8.11 (m, 4H, C6H4), 9.45 (s, 1H,
NH–CH2); 13C-NMR d: 20.2 (quinazolinone–CH3),54.5
(CH2), 120.9, 122.4, 127.4, 128.8, 133.5, 147.1 (benzene),
161.1 (C=O), 164.2 (C–CH3–quinazolinone), 170.3 (C=O);
Anal. Cald. for C11H13N5O2 (247.25): C, 53.43; H, 5.30; N,
28.32 %. Found: C, 53.39; H, 5.28; N, 28.12 %.
1
10.97 %]; H-NMR d: 3.32 (s, 3H, quinazolinone–CH3),
3.91 (s, 2H, CH2), 4.36 (s, 1H, N–NH), 5.79 (s 1H, imid-
azole–NH), 6.35–8.11 (m, 8H, Ar–H); 13C-NMR d: 20.1
(quinazolinone–CH3), 115.3,120.9, 122.4, 123.0, 127.4,
128.8, 133.5, 138.9, 141.1, 147.1 (benzene, imidazole),
161.1 (C=O, quinazolinone), 164.4 (C–CH3–quinazoli-
none); Anal Cald. for C16H15N5O (305.33): C, 66.87; H,
4.95; N, 22.94 %. Found: C, 66.73; H, 4.86; N, 22.94 %.
2-(2-Methyl-4-oxoquinazolin-3(4H)-ylamino)-N0-
phenylacetohydrazide (5b)
Yield: 1.95 g, 60 %, m.p. 135 °C; IR (KBr) t/cm-1
3,400–3,330 (NH), 3,100 (CH-aromatic), 2,895 (CH3),
:
General procedure for the synthesis of 9a or 9b
2,870 (CH2), 1,680 (C=N), 1,675 (C=O); MS m/z (%) 323
1
[M?, 0.01 %]; H-NMR d: 2.24 (s, 3H, quinazolinone–
To a solution of compound 2 (2.61 g, 0.01 mol) in ethanol
(30 mL) containing triethylamine (0.5 mL), either malon-
onitrile (0.66 g, 0.01 mol) or ethyl cyanoacetate (1.13 g,
0.01 mol) was added. The reaction mixture was heated
under reflux for 2 h, and then poured onto ice/water. The
formed solid product in each case was collected by filtra-
tion, dried, and crystallized from ethanol.
CH3), 3.29 (s, 1H, NH–CH2), 4.76 (s, 2H, CH2), 5.12 (s,
1H, NH–Ph), 6.66–7.93 (m, 9H, Ar–H), 8.30 (s, 1H, NH–
NH–C6H5); 13C-NMR d: 20.2 (quinazolinone–CH3),54.5
(CH2),113.2, 119.2, 120.9, 122.4, 127.4, 128.8, 129.3,
133.5,147.1, 151 (benzene), 161.2 (C=O), 164.3 (C–CH3–
quinazolinone), 170.3 (C=O), Anal. Cald. for C17H17N5O2
(323.35): C, 63.15; H, 5.30; N, 21.66 %. Found: C, 63.28;
H, 5.28; N, 21.75 %.
Ethyl 3,5-diamino-1-(2-methyl-4-oxoquinazolin-3(4H)-yl)-
1H-pyrrole-2-carboxylate (9a)
3-(2-Oxo-2H-chromen-3-ylamino)-2-methylquinazolin-
4(3H)-one (6)
Yield: 1.30 g, 49 %, m.p. 285 °C; IR (KBr) t/cm-1: 3,455
(NH2), 3,087 (CH-aromatic), 2,915 (CH3), 1,680 (C=N),
1
1,675 (C=O); MS m/z (%) 327 [M?, 3.23 %]; H-NMR d:
To a solution of compound 2 (2.61 g, 0.01 mol) in dioxane
(30 mL) containing piperidine (0.5 mL), salicylaldehyde
(1.22 g, 0.01 mol) was added. The reaction mixture was
heated under reflux for 3 h, and then poured onto ice/water.
The formed solid product was collected by filtration, dried,
and crystallized from ethanol. Yield: 2.16 g, 68 %, m.p.
149 °C; IR (KBr) t/cm-1: 3,450 (NH), 3,100 (CH-aro-
matic), 2,890 (CH3), 1,680 (C=N), 1,670 (C=O); MS
m/z (%) 319 [M?, 14.36 %]; 1H-NMR d: 2.57 (s, 3H,
1.30 (t, 3H, J = 6.84 Hz, CH3), 2.49 (s, 3H, quinazoli-
none–CH3),4.04 (s, 2H, NH2),4.26 (s, 2H, NH2), 4.29 (q,
2H, J = 6.84 Hz, CH2), 7.39–8.34 (m, 5H, C6H4, pyrrole-
H); 13C-NMR d: 14.1 (CH3), 20.7 (quinazolinone–CH3),
60.9 (CH2), 110.1,115.5, 122.2, 123.1 (pyrrole),
120.9,122.4, 127.4, 128.8, 133.5, 147.1 (benzene),164.5
(C–CH3–quinazolinone), 166.1 (C=O),170 (C=O–quinaz-
olinone); Anal. Cald. for C16H17N5O3 (327.13): C, 58.71;
123

2218
Med Chem Res (2013) 22:2207–2221
N-(2-methyl-4-oxoquinazolin-3(4H)-yl)benzofuran-2-
carboxamide (11)
H, 5.23; N, 21.39 %. Found: C, 58.85; H, 5.18; N,
21.43 %.
Yield: 1.45 g, 45 %, m.p. 153 °C; IR (KBr) t/cm-1: 3,330
(NH), 3,083 (CH-aromatic), 2,890 (CH3), 1,680 (C=N),
Ethyl 3-amino-5-hydroxy-1-(2-methyl-4-oxoquinazolin-
3(4H)-yl)-1H-pyrrole-2-carboxylate (9b)
1
1,675 (C=O); MS m/z (%) 319 [M?, 19.7 %]; H-NMR d
3.19 (s, 3H, quinazolinone–CH3), 7.51–8.16 (m, 8H, Ar–H),
8.26 (s, 1H, furan-H), 8.98 (s, 1H, NH); 13C-NMR d: 20.1
(quinazolinone–CH3),109.0, 111.6, 120.9, 121.0, 122.4,
123.3, 124.7, 127.4, 128.8, 130.5, 133.5, 147.1, 150.0,
158.4 (benzene–furan),157.2 (exocylic–C=O), 161.2 (C=O,
quinazolinone), 164.3 (C–CH3–quinazolinone); Anal. Cald.
for C18H13N3O3 (319.31): C, 67.71; H, 4.10; N, 13.16 %.
Found: C, 67.68; H, 4.08; N, 13.08 %.
Yield: 1.96 g, 63 %, m.p. 276 °C; IR (KBr) t/cm-1: 3,400
(OH), 3,450 (NH2), 3,090 (CH-aromatic), 2,915 (CH3),
2,870 (CH2), 1,680 (C=N), 1,675 (C=O); MS m/z (%) 328
1
[M?, 10.67 %], H-NMR d: 2.45 (s, 3H, quinazolinone–
CH3), 2.58 (t, 3H, J = 6.04 Hz, CH3), 3.31 (s, 2H, NH2),
4.19 (q, 2H, J = 6.04 Hz, CH2), 5.23 (s, 1H, OH),
7.42–8.56 (m, 5H, C6H4, pyrrole-H); 13C-NMR d: 14.1
(CH3), 20.7 (quinazolinone–CH3), 60.9 (CH2), 102.2, 108,
117.7, 118 (pyrrole), 120.9, 122.4, 127.4, 128.8, 133.5,
147.1 (benzene), 164.4 (C–CH3–quinazolinone), 166.2
(C=O), 170 (C=O); Anal. Cald. for C16H16N4O4 (328.12):
C, 58.53; H, 4.91; N, 17.06 %. Found: C, 58.49; H, 5.02;
N, 17.18 %.
2-Chloro-N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-
phenylacrylamide (12a)
Yield: 2.15 g, 63 %, m.p. 170 °C; IR (KBr) t/cm-1: 3,460
(NH), 3,085 (CH-aromatic), 2,895 (CH3), 1,680 (C=N),
1
1,675 (C=O); MS m/z (%) 339 [M?, 8.39 %]; H-NMR d:
3.29 (s, 3H, quinazolinone–CH3), 7.51–8.16 (m, 9H, Ar–
H), 7.68 (s, 1H, C=CH), 8.98 (s, 1H, NH); 13C-NMR d:
19.9 (quinazolinone–CH3),120.9, 122.4, 126.4, 127.4,
128.0, 128.7, 128.8, 133.5, 135.2, 147.1 (benzene), 126.6
(C–Cl), 135.5 (C=CH–Ar), 161.0 (C=O, quinazolinone),
164.0 (C–CH3–quinazolinone), 165.9 (C=O–NH); Anal.
Cald. for C18H14N3O2Cl (339.78): C, 63.63; H, 4.15; N,
12.37 %. Found: C, 63.58; H, 4.26; N, 12.30 %.
3-(2-Aminothiazol-4-ylamino)-2-methylquinazolin-4(3H)-
one (10)
To a solution of compound 3 (2.51 g, 0.01 mol) in ethanol
(30 mL), thiourea (0.76 g, 0.01 mol) was added. The
reaction mixture was heated under reflux for 3 h, and then
excess solvent was distilled off. The formed solid product
was collected by filtration, dried, and crystallized from
ethanol. Yield: 1.79 g, 65 %, m.p. 128 °C; IR (KBr)
t/cm-1: 3,460–3,330 (NH2, NH), 3,090 (CH-aromatic),
2,895 (CH3), 1,680 (C=N), 1,675 (C=O); MS m/z (%) 273
2-Chloro-3-(4-methoxyphenyl)-N-(2-methyl-4-
oxoquinazolin-3(4H)-yl)acrylamide (12b)
1
[M?, 15.8 %]; H-NMR d: 3.28 (s, 3H, quinazolinone–
Yield: 2.23 g, 60 %, m.p. 176 °C; IR (KBr) t/cm-1: 3,460
(NH), 3,085 (CH-aromatic), 2,895 (CH3), 1,680 (C=N),
CH3), 4.23 (s, 2H, NH2), 4.56 (s, 1H, N–NH), 5.89 (s 1H,
thiazole–NH), 6.65–8.45 (m, 4H, C6H4); 13C-NMR d: 20.1
(quinazolinone–CH3), 120.9, 122.4, 127.4, 128.8, 133.5,
147.1 (benzene), 108, 139, 169 (thiazole), 161.3 (C=O,
quinazolinone), 164.2 (C–CH3–quinazolinone); Anal. Cald.
for C12H11N5OS (273.31): C, 52.73; H, 4.06; N, 25.62; S,
11.73 %. Found: 52.81; H, 4.12; N, 25.56; S, 11.58 %.
1
1,675 (C=O); MS m/z (%) 369 [M?, 2.19 %]; H-NMR d:
3.29 (s, 3H, quinazolinone–CH3), 3.82 (s, 3H, CH3), 7.45
(s, 1H, C=CH), 7.71–8.11 (m, 8H, Ar–H), 8.68 (s, 1H,
NH); 13C-NMR d: 19.9 (quinazolinone–CH3), 55.9
(OCH3), 114.2,120.9,122.4, 127.4, 127.5, 127.7, 128.8,
133.5, 147.1, 159.9 (benzene), 126.6 (C–Cl),135.5 (C=CH–
Ar), 161.0 (C=O, quinazolinone), 164.4 (C–CH3–quinaz-
olinone), 165.9 (C=O–NH); Anal. Cald. for C19H16ClN3O3
(369.80): C, 61.71; H, 4.36; N, 11.36 %. Found: C, 61.75;
H, 4.26; N, 11.28 %.
General procedure for the synthesis of 11
and 12a, 12b, 12c
To a solution of compound 3 (2.51 g, 0.01 mol) in dioxane
(30 mL) containing piperidine (0.5 mL), either salicylal-
dehyde (1.22 g, 0.01 mol), benzaldehyde (1.08 g, 0.01
mol), p-methoxybenzaldehyde (1.36 g, 0.01 mol), or
p-chlorobenzaldehyde (1.40 g, 0.01 mol) was added. The
reaction mixture was heated under reflux for 2 h, and then
poured onto ice/water. The formed solid product was col-
lected by filtration, dried, and crystallized from ethanol.
2-Chloro-3-(4-chlorophenyl)-N-(2-methyl-4-
oxoquinazolin-3(4H)-yl)acrylamide (12c)
Yield: 1.98 g, 53 %, m.p. 146 °C; IR (KBr) t/cm-1: 3,460
(NH), 3,085 (CH-aromatic), 2,895 (CH3), 1,680 (C=N),
1,675 (C=O); MS m/z (%) 373 [M?, 0.40 %]; H-NMR d:
3.29 (s, 3H, quinazolinone–CH3), 7.32 (s, 1H, C=CH),
1
123

Med Chem Res (2013) 22:2207–2221
2219
7.31–8.75 (m, 8H, Ar–H), 8.57 (s, 1H, NH); 13C-NMR d:
19.9 (quinazolinone–CH3),120.9, 122.4, 127.4, 127.8,
128.8, 129.0, 133.3, 133.5, 133.8, 147.1 (benzene), 126.6
(C–Cl), 135.5 (C=CH–Ar), 161.0 (C=O, quinazolinone),
164.2 (C–CH3–quinazolinone), 165.9 (C=O–NH); Anal.
Cald. for C18H13N3O2Cl2 (373.04): C, 57.77; H, 3.50; N,
11.23 %. Found: C, 57.84; H, 3.26; N, 11.16 %.
3-(3,4-Dihydro-4,6-dimethyl-3-phenyl-2-thioxopyrimidin-
1(2H)-yl)-2-methylquinazolin-4(3H)-one (15)
To a solution of compound 4 (3.11 g, 0.01 mol) in dioxane
(30 mL) containing piperidine (0.5 mL), ethyl acetoacetate
(1.30 g, 0.01 mol) was added. The reaction mixture was
heated under reflux for 3 h, and then excess solvent was
distilled off. The formed solid product was collected by
filtration, dried, and crystallized from ethanol. Yield:
2.36 g, 63 %, m.p. 232 °C; IR (KBr) t/cm-1: 3,100 (CH-
aromatic), 2,899 (CH3), 1,680 (C=N), 1,675 (C=O),1,245
(C=S); MS m/z (%) 376 [M?, 0.91 %]; 1H-NMR d: 3.36 (s,
3H, quinazolinone–CH3), 3.78 (s, 3H, pyrimidine–CH3),
6.63 (s, 1H, pyrimidine), 6.90–7.57 (m, 9H, Ar–H);
13C-NMR d:15.8 (CH3, pyrimidine), 20.9 (quinazolinone–
CH3), 120.9, 121.6, 122.4, 124.4, 127.4, 128.8, 129.0,
132.8,133.5, 147.1 (benzene), 103,159.5 (pyrimidine),
161.0 (C=O, quinazolinone), 164.2 (C–CH3–quinazoli-
none),165.4 (C=O, pyrimidine), 178 (C=S); Anal. Cald. for
C20H16N4O2S (376.43): C, 63.81; H, 4.28; N, 14.88, S,
8.52 %. Found: C, 63.75; H, 4.25; N, 14.78; S, 8.33 %.
4-[(2-Methyl-4-oxoquinazolin-3(4H)-yl)amino]-3-phenyl-
1,3-thiazol-2(3H)-ylidene]propanedinitrile (13)
Equimolecular amounts of malononitrile (0.66 g,
0.01 mol) and phenylisothiocyanate (1.35 g, 0.01 mol) and
potassium hydroxide (0.56 g, 0.01 mol) were stirred for
24 h. Next, compound 3 (2.51 g, 0.01 mol) was added and
the reaction mixture was stirred for another 24 h. The
reaction mixture was then poured onto an ice/water mixture
containing few drops of hydrochloric acid, and the formed
solid product was collected by filtration and crystallized
from ethanol. Yield: m.p. 140 °C; IR (KBr) t/cm-1: 3,460
(NH), 3,085 (CH-aromatic), 2,895 (CH3), 1,680 (C=N),
1
1,675 (C=O); MS m/z (%) 398 [M?, 20 %]; H-NMR d:
3-(3,4-Dihydro-6-methyl-4-oxo-3-phenyl-2-
thioxopyrimidin-1(2H)-yl)-2 methylquinazolin-4(3H)-one
(16)
4.32 (s, 1H, CH–thiazole), 4.56 (s, 1H, N–NH), 6.31–7.95
(m, 9H, Ar–H), 3.15 (s, 3H, quinazolinone–CH3);
13C-NMR d: 20.3 (quinazolinone–CH3),77 (C=CH–thia-
zole), 116.3,118.8, 128.8, 120.9, 129.6, 141.3, 122.4,
133.5, 127.4,147.1 (benzene), 152.3 (C=CH–thiazole),
179.8 (N–C–S–thiazole), 107.8 (exocyclic C=C), 161.0
To a solution of compound 4 (3.11 g, 0.01 mol) in dioxane
(30 mL) containing piperidine (0.5 mL), acetylacetone
(1.00 g, 0.01 mol) was added. The reaction mixture was
heated under reflux for 3 h, and then excess solvent was
distilled off. The formed solid product was collected by
filtration, dried, and crystallized from ethanol.
(C–CH3–quinazolinone),164.2
(C–CH3–quinazolinone),
113.7 (CN); Anal. Calcd. for C21H14N6OS (398.44): C,
63.30; H, 3.54; N, 21.09; S, 8.05 %. Found: C, 63.17; H,
3.45; N, 20.88; S, 8.13 %.
Yield: 2.10 g, 56 %, m.p. 240 °C; IR (KBr) t/cm-1
:
3,100 (CH-aromatic), 2,899 (CH3), 1,680 (C=N), 1,675
(C=O), 1,245 (C=S); MS m/z (%) 376 [M?, 0.62 %];
1H-NMR d: 6.92–7.55 (m, 9H, Ar–H), 6.59 (d, 1H,
pyrimidine H-5), 4.89 (s, 1H, pyrimidine H-4), 3.80 (s, 3H,
pyrimidine–CH3), 3.68 (s, 3H, pyrimidine–CH3), 3.31 (s,
3H, quinazolinone–CH3); 13C-NMR d: 17.3, 19.7 (2 CH3,
pyrimidine), 20.9 (quinazolinone–CH3), 120.9, 124.8,
128.8, 127.4, 133.5, 122.4, 147.1, 126.5, 129.1, 140.3
(benzene), 54.8, 99.8, 145.3 (pyrimidine), 161.0 (C=O,
quinazolinone), 164.2 (C–CH3–quinazolinone), 178.2
(C=S); Anal. Cald. for C21H20N4OS (376.47): C, 67.00; H,
5.35; N, 14.88; S, 8.52 %. Found: C, 67.09; H, 5.29; N,
14.78; S, 8.49 %.
N-(2-Methyl-4-oxoquinazolin-3(4H)-yl)-3-oxoindoline-2-
carboxamide (14)
To a solution of compound 3 (2.51 g, 0.01 mol) in eth-
anol (30 mL), o-aminobenzoic acid (1.37 g, 0.01 mol) was
added. The reaction mixture was heated under reflux for
1.5 h, and then excess solvent was distilled off. The formed
solid product was collected by filtration, dried, and crys-
tallized from ethanol. Yield: 1.79 g, 54 %, m.p. 139 °C;
Mpt IR (KBr) t/cm-1: 3,450 (NH), 3,095 (CH-aromatic),
2,895 (CH3), 1,680 (C=N), 1,675 (C=O); MS m/z (%) 334
1
[M?, 5.21 %]; H-NMR d: 3.10 (s, 3H, quinazolinone–
CH3), 5.75 (s, 1H, pyrrole-H), 7.41–8.07 (m, 8H, Ar–H),
8.63 (s, 1H, NH); 13C-NMR d: 19.9 (quinazolinone–CH3),
80.4, 113.4, 117.1, 120.6, 120.9, 122.4, 127.4, 128.8,
129.6, 133.5, 134.5, 147.1, 160.6 (benzene, pyrrole),
161.0 (C=O, quinazolinone), 164.3 (C–CH3–quinazoli-
none), 170.3 (C=O–NH), 196.5 (C=O, pyrrole); Anal.
Calcd. for C18H14N4O3 (334.33): C, 64.66; H, 4.22; N,
16.76 %. Found: C, 64.72; H, 4.20; N, 16.82 %.
3-(4-Oxo-3-phenylthiazolidin-2-ylideneamino)-2-
methylquinazolin-4(3H)-one (17)
To a solution of compound 4 (3.11 g, 0.01 mol) in ethanol
(30 mL), ethyl chloroacetate (1.22 g, 0.01 mol) was added.
The reaction mixture was heated under reflux for 3 h, and
123

2220
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then excess solvent was distilled off. The formed solid
product was collected by filtration, dried, and crystallized
from ethanol. Yield: 2.56 g, 73 %, m.p. 185 °C; IR (KBr)
t/cm-1: 3,100 (CH-aromatic), 2,899 (CH3), 2,779 (CH2),
1,680 (C=N), 1,675 (C=O), 1,660 (exocyclic C=N), MS
1
m/z (%) 350 [M?, 1.37 %]; H-NMR d: 3.31 (s, 3H, qui-
nazolinone–CH3), 5.65 (s, 2H, CH2–thiazole), 6.91–7.56
(m, 9H, Ar–H); 13C-NMR d: 20.4 (quinazolinone–CH3),
29.9 (CH2–thiazole), 120.9, 121.6, 122.4, 124.4, 127.4,
128.8, 129.0, 133.5, 135.4, 147.1 (benzene), 148.4 (exo-
cyclic C=N), 160.0 (C=O, quinazolinone), 164.4 (C–CH3–
quinazolinone), 172.9 (C=O, thiazole); Anal. Cald. for
C18H14N4O2S (350.39): C, 61.70; H, 4.03; N, 15.99; S,
9.15 %. Found: C, 61.79; H, 4.12; N, 16.12; S, 9.26 %.
3-(4-Amino-3-phenylthiazol-2(3H)-ylideneamino)-2-
methylquinazolin-4(3H)-one (18)
To a solution of compound 4 (3.11 g, 0.01 mol) in ethanol
(30 mL), bromoacetonitrile (1.20 g, 0.01 mol) was added.
The reaction mixture was heated under reflux for 3 h, and
then excess solvent was distilled off. The formed solid
product was collected by filtration, dried, and crystallized
from ethanol. Yield: 1.92 g, 55 %, m.p. 143 °C; IR (KBr)
t/cm-1: 3,460 (NH2), 3,100 (CH-aromatic), 2,899 (CH3),
2,890 (CH2), 1,680 (C=N), 1,675 (C=O); MS m/z (%) 349
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RPS 7:87–94
1
[M?, 0.43 %]; H-NMR d: 3.30 (s, 3H, quinazolinone–
CH3), 4.43 (s, 2H, NH2), 5.64 (s, 1H, thiazole–H),
6.90–7.56 (m, 9H, Ar–H); 13C-NMR d: 20.4 (quinazoli-
none–CH3), 65 (CH, thiazole), 116.3, 118.8, 123.9, 122.4,
127.4, 128.8, 129.6, 133.5, 141.3, 147.1 (benzene), 154.0
(exocyclic C=N), 160.0 (C=O, quinazolinone), 161.0 (thi-
azole, C), 164.1 (C–CH3–quinazolinone); Anal. Cald. for
C18H15N5OS (349.41): C, 61.87; H, 4.33; N, 20.04; S, 9.18 %.
Found: C, 61.82 H, 4.43; N, 20.12; S, 9.23 %.
´
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structure activity relationship of a series of potent, orally active,
VEGF receptor tyrosine kinase inhibitors.
45:1300–1312
J Med Chem
´
´
Khabnadideh S, Pez D, Musso M, Brun R, Perez LMR, Gonzalez D,
Pacanowska, Gilbert IH (2005) Design, synthesis and evaluation
of 2,4-diaminoquinazolines as inhibitors of trypanosomal and
leishmanial dihydrofolate reductase. Bioorg Med Chem
13:2637–2649
Acknowledgments The authors would like to extend their gratitude
to the American University in Cairo for its continuous support
through funding this research project.
Kornet MJ, Varia T, Beaven W (1983) Synthesis and anticonvulsant
activity of 3-amino-4(3H)-quinazolinones. J Het Chem 20:1553–
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