Efficient one-pot tandem synthesis and cytotoxicity evaluation of 2,3-disubstituted quinazolin-4(3H)-one derivatives

Article abstract of DOI:10.1016/j.tet.2021.132426

Twenty 2,3-disubstituted quinazolin-4(3H)-one derivatives 1–20 were successfully synthesized in moderate to good yields (25–82%). Their syntheses were based on a one pot tandem ring opening procedure followed by iodine-catalyzed oxidative cyclization of isatoic anhydride with aldehydes, using water as the only solvent under both classical and microwave irradiation conditions. Cytotoxicity assays of the prepared compounds against three human cancer cell lines (HeLa, MCF-7, and A549) indicated that 2, 3, and 20 possessed moderate activities against MCF-7 cells (IC₅₀ = 47.2 μM, 43.9 μM, and 44.9 μM, respectively). Good cytotoxic activities against A549 cells were observed for 3 and 8 with IC₅₀ values of 30.7 μM and 29.8 μM, respectively, which were comparable to the positive control, 5-fluorouracil (5-FU, IC₅₀ = 27.9 μM). Furthermore, compound 4 exhibited slightly stronger activity (IC₅₀ = 23.6 μM) than the positive control 5-FU against the A549 cell line.

Full text of DOI:10.1016/j.tet.2021.132426

Tetrahedron xxx (xxxx) xxx
Contents lists available at ScienceDirect
Tetrahedron
journal homepage: www.elsevier.com/locate/tet
Efficient one-pot tandem synthesis and cytotoxicity evaluation of 2,3-
disubstituted quinazolin-4(3H)-one derivatives
,
Hue Thi Buu Bui a **, Kiep Minh Do b, Huy Tran Duc Nguyen a, Hieu Van Mai a,
,
*
Thanh La Duc Danh a, De Quang Tran a, Hiroyuki Morita b
a Department of Chemistry, College of Natural Sciences, Can Tho University, Viet Nam
b Institute of Natural Medicine, University of Toyama, 2630-Sugitani, Toyama, 930-0194, Japan
a r t i c l e i n f o
a b s t r a c t
Article history:
Twenty 2,3-disubstituted quinazolin-4(3H)-one derivatives 1e20 were successfully synthesized in
moderate to good yields (25e82%). Their syntheses were based on a one pot tandem ring opening
procedure followed by iodine-catalyzed oxidative cyclization of isatoic anhydride with aldehydes, using
water as the only solvent under both classical and microwave irradiation conditions. Cytotoxicity assays
of the prepared compounds against three human cancer cell lines (HeLa, MCF-7, and A549) indicated that
Received 21 June 2021
Received in revised form
18 August 2021
Accepted 23 August 2021
Available online xxx
2, 3, and 20 possessed moderate activities against MCF-7 cells (IC50 ¼ 47.2
respectively). Good cytotoxic activities against A549 cells were observed for 3 and 8 with IC50 values of
30.7 M and 29.8 M, respectively, which were comparable to the positive control, 5-fluorouracil (5-FU,
IC50 ¼ 27.9 M). Furthermore, compound 4 exhibited slightly stronger activity (IC50 ¼ 23.6 M) than the
positive control 5-FU against the A549 cell line.
mM, 43.9 mM, and 44.9 mM,
Keywords:
Aldehyde
Amine
Cytotoxicity
Heterocycle
Iodine
m
m
m
m
© 2021 Elsevier Ltd. All rights reserved.
Microwave
One pot synthesis
4(3H)-Quinazolinones
1. Introduction
oxidative cyclocondensation by electrophiles, in one pot three
component reactions has attracted a great deal of attention [6].
Representative examples of these methodologies include oxida-
tion/cyclization reactions of isatoic anhydride with (1) benzyl ha-
lides [7], (2) N,N-dialkyl carbodiimides [8], and (3) orthoesters
[9e14], especially with aromatic aldehydes (Fig. 2). More recently,
different catalytic systems including ceric ammonium nitrate (CAN)
in ethanol [15], CuO nanoparticles [16], Bi(NO3)3$5H2O [17], thia-
mine hydrochloride (VB1) [18], Ga(OTf)3 [19], and L-prolinium ni-
Cancer continues to be a leading health problem worldwide, and
the pursuit of novel clinically useful anticancer agents is therefore
one of the top priorities of drug development research.
Quinazolinone-based structures have been identified in different
classes of cancer chemotherapeutic agents [1]. Several 2,3-
disubstituted quinazolin-4(3H)-one derivatives reportedly possess
antitumor activity [2], and as shown in Fig. 1, the developed de-
rivatives I [3], II [4], and III [5] are representative examples.
Due to the extensive range of applications of 2,3-disubstituted
4(3H)-quinazolinone derivatives as anticancer agents, consider-
able efforts have been made toward the expansion of synthetic
approaches. Among the preparation methods for all known 2,3-
disubstituted 4(3H)-quinazolinone derivatives, the ring opening
of isatoic anhydride by nitrogen nucleophiles, followed by the
trate inside the meso-channels of (a-Fe2O3)-MCM-41 [20] have
been reported (Fig. 2 and 4a). These routes, however, generally
suffer from low yields of products, prolonged reaction times, high
temperatures, and the use of metallic, non-recyclable catalysts.
Iodine is an inexpensive and readily available catalyst for various
organic transformations under very mild and convenient condi-
tions, to afford the corresponding products in excellent yields with
* Corresponding author.
** Corresponding author.
E-mail addresses: btbhue@ctu.edu.vn (H.T.B. Bui), hmorita@inm.u-toyama.ac.jp
(H. Morita).
https://doi.org/10.1016/j.tet.2021.132426
0040-4020/© 2021 Elsevier Ltd. All rights reserved.
Please cite this article as: H.T.B. Bui, K.M. Do, H.T.D. Nguyen et al., Efficient one-pot tandem synthesis and cytotoxicity evaluation of 2,3-
disubstituted quinazolin-4(3H)-one derivatives, Tetrahedron, https://doi.org/10.1016/j.tet.2021.132426

H.T.B. Bui, K.M. Do, H.T.D. Nguyen et al.
Tetrahedron xxx (xxxx) xxx
Fig. 1. Relevant 2,3-disubstituted 4(3H)-quinazolinones with cytotoxic activities.
Fig. 2. General three component reaction pathways towards the production of 2,3-disubstituted 4(3H)-quinazolinones from isatoic anhydride, and the proposed synthetic
procedure.
high selectivity [21]. A three-component condensation of isotoic
anhydride, primary amine, and aldehyde in the presence of iodine
has been successfully developed for the synthesis of 2,3-substituted
4(3H)-quinazolinones [22,23] (Fig. 2 and 4b). However, the use of
toxic organic solvents, and the prolonged reaction times under
strong basic or acidic conditions are the main drawbacks of these
procedures. Thus, the development of a facile and eco-friendly
method towards the synthesis of 2,3-disubstituted 4(3H)-quina-
zolinones is highly desirable.
or catalysts and requiring a shorter reaction time. The starting
materials are also inexpensive and commercially available. Conse-
quently, twenty 2,3-disubstituted 4(3H)-quinazolinones have been
synthesized and evaluated for their cytotoxicities against three
human cancer cell lines, HeLa, MCF-7, and A549.
2. Results and discussion
2.1. Chemistry
As a solvent, water offers many advantages as it is readily
available, inexpensive, non-toxic, and non-flammable, and thus the
use of water as substitute for organic solvent is very attractive from
both economic and environmental points of view [24]. In addition,
the application of microwave dielectric heating for performing
organic reactions has many advantages, such as remarkable re-
ductions of reaction time, improved yields, and cleaner reactions
than those performed with conventional thermal heating [25]. In
an effort to develop an efficient, green synthetic procedure towards
2,3-disubstituted 4(3H)-quinazolinones, herein we report a novel
one pot tandem ring opening of isatoic anhydride followed by an
iodine-catalyzed oxidative cyclization process with water as the
only solvent, using microwave heating in the ring forming step. This
method offers several advantages, such as omitting toxic solvents
To synthesize 2,3-disubstituted 4(3H)quinazolinones, we
attempted the one pot tandem ring opening of isatoic anhydride by
a primary amine, followed by iodine-catalyzed oxidative cyclization
with aldehyde, using water as the only solvent (Scheme 1).
First, the formation of 2,3-disubstituted-3H-quinazolin-4-one 4
was explored using isatoic anhydride 1, benzylamine 2, and benz-
aldehyde 3 as the model substrates, in the presence of K2CO3 in
water and iodine as the oxidant. The reactions were performed
using different molar ratios of reactants, as presented in Table 1
(Entries 1e11) [22]. The best yield (80%) of the desired product 4
was obtained by applying the reaction conditions shown in Entry 7.
The efficiency of the reaction significantly decreased when K2CO3
was replaced with other bases, such as KI or NaOMe (Table 1,
2

H.T.B. Bui, K.M. Do, H.T.D. Nguyen et al.
Tetrahedron xxx (xxxx) xxx
Scheme 1. One pot tandem synthesis of 2,3-disubstituted 4(3H)quinazolinones in water.
Table 1
amine or ammonium acetate, and aldehydes, followed by an
oxidation reaction using iodine as the oxidant [22,23]. However,
our present method has several distinguishing features that are
worth mentioning: (i) it is environmentally friendly and cost-
effective by utilizing water as the solvent under mild basic condi-
tions, (ii) it proceeds faster, within 30 min, under microwave irra-
diation, (iii) it is applicable to various substituted benzylamine and
aromatic aldehydes.
Optimization of the reaction conditionsa.
Entry Amine (eq) Aldehyde (eq) Oxidant (eq) Base (eq)
Yield (%)b
2.2. Cytotoxicity evaluation
1
2
3
4
5
6
7
8
1.0
1.05
1.05
1.1
1.1
1.1
1.1
1.5
1.3
1.1
1.1
1.1
1.1
1.1
1.0
1.05
1.10
1.1
1.1
1.1
1.1
1.1
1.1
1.1
1.1
1.1
1.1
1.1
I2 (1.0)
K2CO3 (1.0)
K2CO3 (2.0)
K2CO3 (2.0)
K2CO3 (2.0)
K2CO3 (1.0)
K2CO3 (1.1)
K2CO3 (1.2)
K2CO3 (2.0)
K2CO3 (2.0)
K2CO3 (1.2)
K2CO3 (1.3)
KI (1.2)
69
38
62
65
76
78
80
46
60
64
51
68
I
I
I
I
I
2 (1.5)
2 (1.5)
2 (1.5)
2 (1.0)
2 (1.1)
The cytotoxicities of the prepared 2,3-disubstituted 4(3H)-qui-
nazolinones 1e20 against three human cancer cell lines (HeLa,
MCF-7, and A549) were examined (Table 3). The results indicated
that all tested compounds, except for 1, 4e6, 12e14, and 17, showed
weak cytotoxicities against the HeLa cell line. The presence of a free
hydroxy group at either the N-3 (compounds 5, 6, 12) or the C-2
(compound 14) position was not effective for enhancing the cyto-
toxicities against both the MCF-7 and A549 cell lines. Similar results
were also observed for the CH3 (compound 7 and 15) or 3,4,5-
trimethoxyphenyl (compound 13) substituents. In contrast, the
introduction of an electron donating group such as OCH3 (com-
pound 2) or an electron withdrawing group such as F (compound 3)
on the benzene ring at the N-3 position of the quinazolinone
structure enhanced the activity against both the MCF-7
I2 (1.2)
I
I
I
I
I
I
I
2 (1.5)
2 (1.5)
2 (1.3)
2 (1.2)
2 (1.2)
2 (1.2)
2 (1.2)
9
10
11
12
13
14
NaOMe (1.2) 53
51
e
a
Reaction conditions: a mixture of isatoic anhydride 1 (0.3 mmol), benzylamine
2a, and base in water (4 mL) was stirred at RT for 30 min, and then benzaldehyde 3a
was added. The mixture was stirred for 5 additional min, and then the oxidant was
added. The resulting mixture was stirred at 80 ꢀC for 4 h.
b
Isolated yields.
(IC50 ¼ 47.2
A549 cell lines (IC50 ¼ 48.3 4.03
tively), as compared to the unsubstituted derivative
(IC50 > 100 M). Notably, the presence of fluorine (F) on the ben-
1.70
mM and 43.9
0.49
m
M, respectively) and
m
M and 30.7 0.91
mM, respec-
1
Entries 12, 13) or in the absence of a base (Entry 14). In addition, no
reaction occurred when iodine was absent in the reaction or other
oxidants including Oxone or Na2S2O5 were used as substitutes for
iodine.
Having established the optimum reaction conditions, the scope
of the reaction was studied by using various substituted benzyl- or
alkylamines and substituted aromatic aldehydes without any
modifications. As shown in Table 2, the reaction conditions were
applicable to a broad range of substituted benzylamines and aro-
matic aldehydes, leading to the formation of 19 corresponding 2,3-
disubstituted-3H-quinazolin-4-ones 2e19 in moderate to good
yields.
In an effort to shorten the reaction time, microwave heating was
applied for the ring forming step. Thus, after treating isatoic an-
hydride with an amine, followed by an aromatic aldehyde, iodine
was subsequently added and the resulting mixture was subjected
to microwave irradiation. As a result, almost all the reactions were
accomplished within 30 min, and slight increases in the yields of
the desired products were observed as compared to the classical
heating method (Table 2). The presence of a free hydroxy group is
not tolerated by the present synthetic method under both classical
and microwave heating conditions (Table 2, Entries 5, 6, and 12).
The 2,3-disubstituted quinazolinones have previously been syn-
thesized via a one pot condensation of isatoic anhydride, primary
m
zene ring (compound 3) resulted in cytotoxicity at a level consistent
with that of 5-fluorouracil (5-FU) against the A549 cell line
(IC50 ¼ 27.9 3.90
mM). Interestingly, although the incorporation of
the 1,3-dioxolyl group at this position (compound 4) decreased the
activity on the MCF-7 cells, it considerably increased the cytotox-
icity against the A549 cells (IC50 ¼ 23.6 5.30
slightly stronger activity relative to the positive control 5-FU
M). In contrast, further functionalization of 4
mM), and exhibited
(IC50 ¼ 27.9 3.90
m
by adding a Cl group on the benzene ring at the C-2 position
(compound 20) induced moderate activity against the MCF-7 cells
(IC50 ¼ 44.9 3.73
mM), but significantly reduced the activity on the
A549 cell line.
In general, for the A549 cell line, it seems that the methoxy
(compound 2) or the fluorine (compound 3), and especially the 1,3-
dioxolyl group (compound 4), in the benzyl moiety at the N-3 po-
sition of the quinazolinone system effectively induced the cyto-
toxicity only when associated with an unsubstituted phenyl group
at the C-2 position (Fig. 3). Similarly, the methoxy in the benzene
ring at the C-2 position (compound 8) only achieves the activity
when it is in conjunction with the unsubstituted benzyl group at
the N-3 position. The simultaneous incorporation of these sub-
stituents at both the N-3 and C-2 positions likely leads to the loss of
the activity. The structural features of compounds 2 and 3, or the
3

H.T.B. Bui, K.M. Do, H.T.D. Nguyen et al.
Tetrahedron xxx (xxxx) xxx
Table 2
Syntheses of 2,3-disubstituted 4(3H)-quinazolinones via one pot tandem reactions in water.
4

H.T.B. Bui, K.M. Do, H.T.D. Nguyen et al.
Tetrahedron xxx (xxxx) xxx
Table 3
the activity, which will be reported in due course.
Cytotoxic activities of compounds 1e20.
4. Experimental section
Samples
IC50 (mM)
HeLa
MCF-7
A549
4.1. General information
1
>100
>100
>100
2
3
4
5
6
7
8
9
85.1 1.00
85.3 0.72
>100
>100
>100
81.2 1.04
93.8 1.06
63.4 7.88
69.9 2.86
83.2 2.37
>100
47.2 1.70
43.9 0.49
90.8 3.68
>100
>100
>100
72.7 1.61
>100
70.1 0.06
50.8 1.25
>100
48.3 4.03
30.7 0.91
23.6 5.30
>100
>100
>100
29.8 2.42
80.8 5.16
43.9 3.60
67.0 2.20
>100
Reactions were monitored by thin-layer chromatography (TLC)
on 0.2 mm pre-coated silica-gel 60 F254 plates (Merck). 1H NMR
and 13C NMR spectra were measured with Bruker Avance 500 and
600 MHz spectrometers. HRESI-MS observations were performed
on a Sciex OS 1.2 mass spectrometer. FT-IR was conducted using the
KBr pellet method on a Thermo Nicolet 6700 spectrometer.
Chemical shifts are given in parts per million (ppm) relative to
10
11
12
13
14
15
16
17
18
19
20
5-FU*
tetramethylsilane (Me4Si,
d
¼ 0); J values are given in Hertz. All
chemicals and solvents used in this study were of analytical grade.
Reactions under microwave irradiation were performed in a mi-
crowave synthesis system (CEM Discovery).
>100
>100
>100
>100
>100
>100
88.1 0.66
91.7 1.04
>100
84.6 0.15
86.0 5.43
96.3 4.10
29.9 1.63
67.6 1.20
57.5 1.64
>100
74.4 2.78
>100
92.1 4.22
86.0 2.07
92.4 3.32
84.9 3.60
84.4 0.75
89.2 3.42
27.9 3.90
4.2. General method for the synthesis of 2,3-disubstituted 4(3H)-
quinazolinones
44.9 3.73
35.4 4.56
4.2.1. Classical conditions
A mixture of isatoic anhydride (0.3 mmol), amine (0.33 mmol),
and K2CO3 (0.36 mmol) in H2O (4 mL) was stirred at room tem-
Data are presented as mean SD (n ¼ 3).
*5-Fluorouracil: positive control.
Fig. 3. Important structural features for cytotoxicity against the A549 cell line.
combined structure of the 1,3-dioxolyl group in the benzyl moiety
at the N-3 and the Cl group in the benzene ring at the C-2 position
(compound 20), seem to be critical for the cytotoxicity against the
MCF-7 cell line. Similar cases were also found in the previous
studies, where the presence of either fluorine, chlorine, or methoxy
groups in the quinazolinone ring have been indicated to be
important for enhancing cytotoxicity of quinazolinones against
MCF-7 and/or A549 cell lines [36].
perature for 30 min, and aldehyde (0.33 mmol) was then added.
The resulting mixture was stirred for 5 more min, followed by the
addition of I2 (0.36 mmol). The reaction mixture was stirred at 80 ꢀC
for 4 h and quenched with a saturated aqueous solution of Na2S2O3
(4 mL). The aqueous layer was extracted with EtOAc (3 ꢁ 20 mL).
The combined organic layers were washed with a saturated
aqueous solution of NaCl and dried with Na2SO4, and then the
solvent was evaporated under reduced pressure to obtain the crude
product. Purification by column chromatography on silica gel, using
a mixture of hexane and ethyl acetate as solvents, afforded the
desired products 1e20. All compounds were 95%þ pure according
to high resolution 500 MHz 1H NMR, see spectra in Supporting
Information.
3. Conclusions
A simple, environmentally friendly, one pot tandem procedure
has been developed for the synthesis of 2,3-disubstituted-3H-qui-
nazolin-4-one under mild conditions via the cyclocondensation of
isatoic anhydride with a primary amine and aldehyde in water, and
iodine as the oxidant, using microwave heating in the ring forming
step. This offers several advantages, such as moderate to high
yields, simple procedure, low cost, short reaction times, and mild
and organic solvent-free conditions. The cytotoxicity evaluations of
the prepared compounds against three human cancer cell lines
(HeLa, MCF-7, and A549) showed that compounds 2 and 3 both
possessed moderate activities against the MCF-7 and A549 cancer
cell lines, while compound 20 moderately affected only the MCF-
7 cell line. Remarkably, compounds 4 and 8 showed cytotoxicities
comparable to that of the positive control on the A549 cell line.
Studies ongoing in our lab include monitoring the effects of other
pharmacophoric groups at different positions in the quinazolinone
system on the cytotoxicity, as well as elucidating the mechanism of
4.2.2. Microwave conditions
A mixture of isatoic anhydride (0.3 mmol), amine (0.33 mmol),
and K2CO3 (0.36 mmol) in H2O (4 mL) was stirred at room tem-
perature for 30 min, and aldehyde (0.33 mmol) was then added.
The resulting mixture was stirred for 5 more min, followed by
addition of I2 (0.36 mmol). The reaction mixture was subjected to
microwave irradiation conditions for 30 min (CEM Discovery
reactor), using the Power Cycling mode with specific parameters as
follows: power: 100 W; power interval: 15 s; cooling interval:
2 min; max temp: 250 ꢀC; min temp:100 ꢀC; number of cycles: 20.
The workup procedure was the same as that described for the
synthesis under classical conditions. Spectral data of compounds
2e6 [26e30], 7e8 [27], 9 [31], 12 [32], 13 [27,33], 15 [22], 16 [27,34],
and 18 [35] were previously described in the literature and are
5

H.T.B. Bui, K.M. Do, H.T.D. Nguyen et al.
Tetrahedron xxx (xxxx) xxx
provided in the Supporting Information.
4.2.8. 2-(4-Chlorophenyl)-3-(4-fluorobenzyl)quinazolin-4(3H)-one
(19)
Yield 64% as yellow solid; mp 136e138 ꢀC; FT-IR (KBr) nmax
(cmꢂ1): 2924, 2852, 1687, 1653, 1601, 1509, 1473, 1380, 1227, 1089,
1027, 861, 775; HRESIMS: m/z 365.0858 [MþH]þ (calcd. for
4.2.3. 3-Benzyl-2-phenylquinazolin-4(3H)-one (1)
Yield 82% as white solid; mp 141e143 ꢀC; FT-IR (KBr) nmax
(cmꢂ1): 3059, 3033, 2958, 2929, 1677, 1494, 1474, 1378, 1245, 1149,
1075, 966, 773, 704; HRESIMS: m/z 313.1341 [MþH]þ (calcd. for
C
21H14ClFN2O 365.8044); 1H NMR (500 MHz, DMSO‑d6)
d: 8.22 (dd,
21H16N2O 313.3720); 1H NMR (500 MHz, DMSO‑d6)
d: 8.22 (dd,
J ¼ 8.0, 1.0 Hz, 1H), 7.87e7.90 (m, 1H), 7.72 (dd, J ¼ 8.0, 0.5 Hz, 1H),
C
7.59e7.63 (m, 1H), 7.46e7.52 (m, 4H), 7.03e7.07 (m, 2H), 6.96e6.98
J ¼ 8.0, 1.0 Hz, 1H), 7.87e7.90 (m, 1H), 7.72 (d, J ¼ 8.0 Hz, 1H),
7.58e7.62 (m, 1H), 7.48e7.51 (m, 1H), 7.41e7.46 (m, 4H), 7.19e7.24
(m, 3H), 6.91 (dd, J ¼ 8.5, 2.0 Hz, 2H), 5.19 (s, 2H). 13C NMR
(m, 2H), 5.15 (s, 2H); 13C NMR (125 MHz, DMSO‑d6)
d: 162.1, 161.3,
160.2, 155.0, 146.8, 134.7, 134.5, 133.9, 132.7, 132.7, 129.9, 128.5,
128.4, 128.3, 127.3, 126.4, 120.4, 115.3, 115.1, 47.4.
(125 MHz, DMSO‑d6) d: 161.4, 156.1, 146.9, 136.7, 135.1, 134.7, 129.6,
128.4, 128.2, 127.9, 127.3, 127.2, 127.0, 126.4, 126.2, 120.3, 48.1.
4.2.9. 3-(Benzo[d] [1,3]dioxol-5-ylmethyl)-2-(4-chlorophenyl)
quinazolin-4(3H)-one (20)
4.2.4. 3-(4-Fluorobenzyl)-2-(4-methoxyphenyl)quinazolin-4(3H)-
one (10)
Yield 58% as white solid; mp 150e152 ꢀC; FT-IR (KBr) nmax
(cmꢂ1): 2923, 2853, 1680, 1601, 1499, 1483, 1446, 1369, 1251, 1088,
1039, 960, 926, 780; HRESIMS: m/z 391.0849 [MþH]þ (calcd. for
Yield 67% as white solid; mp 101e102 ꢀC; FT-IR (KBr) nmax
(cmꢂ1): 3020, 2956, 2927, 2835, 1681, 1607, 1587, 1515, 1379, 1254,
1222, 1034, 770, 691; HRESIMS: m/z 361.1355 [MþH]þ (calcd. for
C
22H15ClN2O3 391.8230); 1H NMR (500 MHz, DMSO‑d6)
d: 8.22 (dd,
C
22H17FN2O2 361.3884); 1H NMR (500 MHz, DMSO‑d6)
d: 8.19 (dd,
J ¼ 8.0, 1.5 Hz, 1H), 7.88 (td, J ¼ 8.5, 1.5 Hz, 1H), 7.71 (d, J ¼ 8.0 Hz,
1H), 7.57e7.62 (m, 1H), 7.49e7.54 (m, 4H), 6.74 (d, J ¼ 8.0 Hz, 1H),
6.51 (d, J ¼ 1.5 Hz,1H), 6.33 (dd, J ¼ 8.0, 2.0 Hz,1H), 5.95 (s, 2H), 5.08
J ¼ 8.0, 1.5 Hz, 1H), 7.86e7.90 (m, 1H), 7.71 (d, J ¼ 8.0 Hz, 1H),
7.57e7.61 (m, 1H), 7.42 (dd, J ¼ 7.0, 2.0 Hz, 2H), 7.07 (t, J ¼ 9.0 Hz,
2H), 6.97e7.01 (m, 4H), 5.20 (s, 2H), 3.82 (s, 3H); 13C NMR
(125 MHz, DMSO‑d6) d: 161.5, 160.1, 155.9, 147.0, 134.6, 133.0, 129.6,
128.4, 128.3, 127.4, 127.2, 127.0, 126.3, 120.2, 125.2, 115.1, 113.6, 55.3,
47.6.
(s, 2H); 13C NMR (125 MHz, DMSO‑d6)
d: 161.3, 155.0, 147.3, 146.8,
146.3, 134.7, 134.4, 133.9, 130.3, 130.0, 128.3, 127.3, 126.4, 120.4,
119.6, 108.1, 107.1, 100.9, 47.8.
4.2.5. 3-(Benzo[d] [1,3]dioxol-5-ylmethyl)-2-(4-methoxyphenyl)
quinazolin-4(3H)-one (11)
4.3. Cytotoxicity evaluation
Yield 54% as yellow solid; mp 130e132 ꢀC; FT-IR (KBr) nmax
(cmꢂ1): 2962, 2913, 2841, 1676, 1608, 1514, 1500, 1484, 1444, 1244,
1034, 781; HRESIMS: m/z 387.1345 [MþH]þ (calcd. for C23H18N2O4
4.3.1. Sample preparation
All synthesized compounds were dissolved in DMSO to make
10 mM stock solutions. Serial dilutions were prepared in culture
medium. The positive control, 5-FU, was dissolved in DMSO to
make a 10 mM stock solution and then stored at ꢂ20 C until use.
387.4070); 1H NMR (500 MHz, DMSO‑d6)
d
: 8.19 (dd, J ¼ 8.0, 1.0 Hz,
1H), 7.84e7.87 (m, 1H), 7.69 (d, J ¼ 8.0 Hz, 1H), 7.55e7.58 (m, 1H),
7.44 (dd, J ¼ 7.0, 2.0 Hz, 2H), 7.09 (dd, J ¼ 6.0, 2.0 Hz, 2H), 6.75 (d,
J ¼ 8.0 Hz, 1H), 6.52 (d, J ¼ 1.5 Hz, 1H), 6.35 (dd, J ¼ 8.0, 2.0 Hz, 1H),
5.95 (s, 2H), 5.12 (s, 2H), 3.81 (s, 3H); 13C NMR (125 MHz, DMSO‑d6)
ο
4.3.2. MTT proliferation assay
d
: 161.5, 160.1, 155.9, 147.2, 146.9, 146.2, 134.6, 130.6, 129.7, 127.5,
The cytotoxic activities of the synthesized compounds were
evaluated against human cancer cell lines (HeLa, MCF-7, and A549),
using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium
bromide (MTT) assay with some modifications [37]. The human
127.2, 126.9, 126.3, 120.0, 119.6, 113.6, 108.1, 107.1, 100.9, 55.3, 47.9.
4.2.6. 3-Benzyl-2-(4-hydroxyphenyl)quinazolin-4(3H)-one (14)
Yield 40% as slightly yellow solid; mp 197e199 ꢀC; FT-IR (KBr)
nmax (cmꢂ1): 3306, 3068, 1647, 1611, 1587, 1578, 1566, 1518, 1472,
1431, 1361, 1276, 1208, 1171, 772; HRESIMS: m/z 329.1288 [MþH]þ
cancer cell lines were cultured in a-minimum essential medium (a-
MEM), supplemented with 1% antibiotic antimycotic solution and
10% fetal bovine serum, at 37 οC and in a 5% CO2 atmosphere. Cells
at 80e90% confluence were harvested and centrifuged at 3000 rpm
for 3 min. The supernatant was discarded and the cell pellet was
(calcd. for C21H16N2O2 329.3710); 1H NMR (500 MHz, DMSO‑d6)
d:
9.89 (s, 1H), 8.18 (dd, J ¼ 8.0, 1.0 Hz, 1H), 7.84e7.87 (m, 1H), 7.69 (dd,
J ¼ 8.0, 0.5 Hz, 1H), 7.54e7.58 (m, 1H), 7.28e7.30 (m, 2H), 7.19e7.25
(m, 3H), 6.94 (d, J ¼ 8.5 Hz, 2H), 6.78 (dt, J ¼ 9.5, 3.0 Hz, 2H), 5.23 (s,
resuspended in fresh medium. Aliquots (100 mL) of the cells were
seeded in 96-well plates (1 ꢁ 104 cells/well) and incubated for 24 h.
The cells were then washed with phosphate-buffered saline (PBS),
and various concentrations of tested compounds, including the
2H); 13C NMR (125 MHz, DMSO‑d6,
d ppm): 161.6,158.6,156.3,147.0,
136.9, 134.6, 129.7, 128.3, 127.2, 127.0, 126.8, 126.3, 126.2, 125.9,
120.1, 114.8, 48.3.
positive control, 5-FU (5e100
m
M), were added to the wells. After a
72 h incubation, the cells were washed with PBS, and 100
mL ali-
4.2.7. 3-(Benzo[d] [1,3]dioxol-5-ylmethyl)-2-(4-fluorophenyl)
quinazolin-4(3H)-one (17)
quots of medium containing MTT solution (5 mg/mL) were added to
each well and incubated for 3 h. The absorbance was recorded using
a microplate reader at 570 nm. Percent proliferation inhibition was
calculated using the following formula:
% Proliferation cell inhibition ¼ [(At ꢂ Ab)/(Ac ꢂ Ab)] ꢁ 100.
At: Absorbance of test compound, Ab: Absorbance of blank, Ac:
Absorbance of control.
The concentrations (IC50 values) of the compounds required to
inhibit 50% of the growth of the human cancer cell lines were
calculated based on the relationship between concentrations and
percent inhibitions, using the GraphPad Prism 5.0 software. Each
experiment was performed three times, and all data are presented
as mean standard deviation (SD).
Yield 60% as white solid; mp 152e154 ꢀC; FT-IR (KBr) nmax
(cmꢂ1): 3057, 2919, 1668, 1604, 1583, 1541, 1507, 1471, 1372, 1332,
1245, 1120, 1093, 1037, 891, 777; HRESIMS: m/z 375.1145 [MþH]þ
(calcd. for C22H15FN2O3 375.3714); 1H NMR (500 MHz, DMSO‑d6)
d:
8.22 (dd, J ¼ 8.0, 1.0 Hz, 1H), 7.86e7.89 (m, 1H), 7.70 (d, J ¼ 8.0 Hz,
1H), 7.58e7.61 (m, 1H), 7.51e7.54 (m, 1H), 7.29 (t, J ¼ 8.5 Hz, 2H),
6.73 (d, J ¼ 8.0 Hz, 1H), 6.50 (d, J ¼ 2.0 Hz, 1H), 6.31 (dd, J ¼ 8.0,
6.0 Hz, 1H), 5.95 (s, 2H), 5.08 (s, 2H); 13C NMR (125 MHz, DMSO‑d6)
d
: 163.5, 161.6, 161.4, 155.2, 147.3, 146.8, 146.3, 134.7, 131.7, 131.7,
130.6, 130.5, 130.4, 127.3, 127.2, 126.4, 120.4, 119.7, 115.3, 115.1, 108.1,
107.1, 100.9, 47.8.
6

H.T.B. Bui, K.M. Do, H.T.D. Nguyen et al.
Tetrahedron xxx (xxxx) xxx
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Declaration of competing interest
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The authors declare that they have no known competing
financial interests or personal relationships that could have
appeared to influence the work reported in this paper.
Acknowledgments
This research was funded by the Vietnam National Foundation
for Science and Technology Development (NAFOSTED)
(104.01e2018.51) and supported in part by a Grant-in-Aid for Sci-
entific Research from the Ministry of Education, Culture, Sports,
Science and Technology, Japan (JSPS KAKENHI Grant, JP19H04649).
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7