Synthesis and antiproliferative evaluation of oxime, methyloxime, and amide-containing quinazolinones

Article abstract of DOI:10.1002/jccs.201700463

Certain oxime, methyloxime, and amide-containing quinazolinone derivatives were synthesized and evaluated in vitro for their antiproliferative activities against a panel of human cancer cell lines including nasopharyngeal carcinoma (NPC-TW01), lung carcinoma (NCI-H226), and leukemia (Jurkat). Quinazolinone 2 was inactive against all three cell lines tested, while quinazolinone 4 was weakly active against both Jurkat and H226 cancer cells with IC₅₀ values of 6.55 and 12.27 μM, respectively, indicating that the oxime derivative 4 is more favorable than its ketone precursor 2. Our results have also indicated that quinazolinone 8g and its biphenyl counterpart 8f exhibited more potent antiproliferative activities than the positive control methotrexate against all three cancer cell lines tested. Among these quinazolinone derivatives, 8g was the most active against NPC-TW01 with an IC₅₀ value of 4.78 μM. Further study on NPC-TW01 cell cycle distribution indicated that the compound 8g induced cell arrest at the G1/G0 phase in a time- and concentration-dependent manner. Moreover, a characteristic hypo-diploid DNA content peak (sub-G1) was found to increase from 1 to 4% in NPC-TW01 cells treated with 8g for 72 hr. These results indicate that 8g can induce cells arrest in the G1/G0 phase and cause cell death. Further structural optimization of 8g and detailed study of its antiproliferative mechanism are going on.

Full text of DOI:10.1002/jccs.201700463

Received: 20 December 2017
DOI: 10.1002/jccs.201700463
Revised: 31 March 2018
Accepted: 19 April 2018
A R T I C L E
Synthesis and antiproliferative evaluation of oxime, methyloxime,
and amide-containing quinazolinones
Ken-Ming Chang1† | Li-Chai Chen1,2† | Cherng-Chyi Tzeng3 | Yao-Hua Lu1 | I-Li Chen1 |
Shin-Hun Juang1,4 | Tai-Chi Wang1
1Department of Pharmacy, Tajen University,
Certain oxime, methyloxime, and amide-containing quinazolinone derivatives were
Pingtung, Taiwan
2Clinical Pharmacy Division, ZuoYing Branch of
Kaohsiung Armed Forces General Hospital,
Kaohsiung, Taiwan
3Department of Medicinal and Applied Chemistry,
College of Life Science, Kaohsiung Medical
University, Kaohsiung, Taiwan
synthesized and evaluated in vitro for their antiproliferative activities against
a panel of human cancer cell lines including nasopharyngeal carcinoma
(NPC-TW01), lung carcinoma (NCI-H226), and leukemia (Jurkat). Quinazolinone
2 was inactive against all three cell lines tested, while quinazolinone 4 was weakly
active against both Jurkat and H226 cancer cells with IC50 values of 6.55 and
12.27 μM, respectively, indicating that the oxime derivative 4 is more favorable
than its ketone precursor 2. Our results have also indicated that quinazolinone 8g
and its biphenyl counterpart 8f exhibited more potent antiproliferative activities
than the positive control methotrexate against all three cancer cell lines tested.
Among these quinazolinone derivatives, 8g was the most active against
NPC-TW01 with an IC50 value of 4.78 μM. Further study on NPC-TW01 cell
cycle distribution indicated that the compound 8g induced cell arrest at the G1/G0
phase in a time- and concentration-dependent manner. Moreover, a characteristic
hypo-diploid DNA content peak (sub-G1) was found to increase from 1 to 4% in
NPC-TW01 cells treated with 8g for 72 hr. These results indicate that 8g can
induce cells arrest in the G1/G0 phase and cause cell death. Further structural
optimization of 8g and detailed study of its antiproliferative mechanism are
going on.
4School of Pharmacy, China Medical University,
Taichung, Taiwan
Correspondence
Tai-Chi Wang, Department of Pharmacy, Tajen
University, Pingtung 907, Taiwan.
Email: tcwang@tajen.edu.tw
Funding information
Ministry of Science and Technology of the
Republic of China, Grant/Award Number:
NSC91-2320-B-127-005
KEYWORDS
antiproliferative activities, nasopharyngeal carcinoma (NPC), quinazolinones
result, the impact of NPC on the family and social stability is
quite devastating. Although pre-radiation chemotherapy may
1
| INTRODUCTION
Nasopharyngeal carcinoma (NPC), a head and neck cancer
that occurs between the top rear area of the throat and nose, is
a rare cancer in Western society but is extremely common in
Southeast Asia and Taiwan.[1–5] In Taiwan, NPC is the tenth
leading cause of mortality in male cancer patients and the most
common among males between 35 and 50 years of age.[6]
Despite an initial response to chemotherapy, the majority of
patients with advanced NPC succumb to this disease.[7,8] As a
enhance the 5-year survival rates of NPC patients, the develop-
ment of drug resistance often results in the failure of treat-
ments. Besides, therapeutic outcome is still unsatisfactory for
patients with refractory and relapsed NPC even in spite of
receiving a second line of docetaxel-based chemotherapy.[9]
Therefore, novel therapeutic drugs and protocols are urgently
needed to overcome drug resistance in NPC patients.
For the past few years, we have been interested in the
design and synthesis of novel heterocyclic compounds as
†These authors contributed equally to this study.
© 2018 The Chemical Society Located in Taipei & Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
J Chin Chem Soc. 2018;1–9.
http://www.jccs.wiley-vch.de
1

2
CHANG ET AL.
potential anti-NPC agents.[10–13] Among them, N-(biphenyl-
4-yl)-2-(2-oxo-1,2-dihydroquinolin-7-yloxy)acetamide (I)[10]
quinazolinone 3a in 90% yield. Accordingly, compounds
3b–3g were prepared from 1 and arylacyl bromide under the
same reaction conditions.
and
N-(naphthalen-2-yl)-2-(4-oxo-2-phenyl-4H-chromen-
7-yloxy)acetamide (II)[11] were found to be selectively
active against the growth of NPC-TW01 with IC50 values of
<10 and 1.37 μM, respectively. More recently, we have pre-
pared certain N-(naphthalen-2-yl)acetamide derivatives
and evaluated in vitro for their antiproliferative activities
against a panel of human cancer cell lines. Among them,
N-(naphthalen-2-yl)-2-(2-oxo-1,2,3,4-tetrahydroquinolin-6-
yloxy)acetamide (III)[12] and 2-(9,10-dioxo-9,10-dihy-
droanthracen-2-yloxy)-N-(naphthalen-2-yl)acetamide (IV)[13]
were found to have selective antiproliferative activities against
NPC-TW01 with IC50 values of 0.6 and 2.62 μM, respec-
tively. In continuation of our studies to explore active and
selective anti-NPC agents and establish structure–activity rela-
tionships, the present report describes the preparation of certain
amide-containing quinazolinone derivatives (Figure 1). Our
selection of quinazolinone derivatives as target compounds is
based on the fact that the quinazolinone ring is an important
and versatile motif of many anticancer drugs, especially epi-
dermal growth factor receptor (EGFR) inhibitors such as
gefitinib, erlotinib, and lapatinib.[14–17] The oxime-containing
quinazolinone derivatives were also synthesized for antiproli-
ferative evaluation.
Treatment of quinazolinone 2 with NH2OH afforded
exclusively quinazolinone 4 in 75% yield. The configuration
of the oxime moiety was determined by through-space
nuclear Overhauser effect spectroscopy (NOESY), which
revealed a coupling connectivity to CH3 protons. The stereo-
specific oximination to give the E-form product rather than
the Z-isomer can be realized in which the hydroxyl group
proximate to the methyl group is less sterically hindered.[18]
Accordingly, quinazolinone 5 was prepared from 2 and
NH2OMe under the same reaction condition. Reaction of
quinazolinone 3a with NH2OH afforded exclusively quina-
zolinone 6a in 88% yield. The configuration of the oxime
moiety was determined by its 13C-NMR spectrum in which
the N-CH2 signal shifted upfield from 52.32 ppm (3a) to
42.19 ppm (6a).[19] The same synthetic procedures were
applied for the synthesis of (Z)-6b–g from 3b–g and (Z)-7a–
g from 3a–g.
The preparation of amide-containing quinazolinone via
Schmidt rearrangement is also illustrated in Scheme 1.
Quinazolinone 3a was treated with H2SO4 and NaN3 to
afford quinazolinone 8a in 90% yield. The same synthetic
procedures were applied for the synthesis of 8b–8g. The
structures of the newly synthesized compounds were con-
firmed by NMR spectra and elementary analysis.
2
| RESULTS AND DISCUSSION
2.2 | Biological activity
2.1 | Synthesis
All compounds were evaluated in vitro against a three-cell-
line panel consisting of NPC-TW01 (human nasopharyngeal
carcinoma), Jurkat (leukemia), and non-small-cell lung car-
cinoma (NCI-H226). Results from Table 1 indicate that qui-
nazolinone 2 is inactive against all three cell lines tested,
while its phenyl analog 3a was weakly active against Jurkat
cancer cells with an IC50 of 8.79 μM. Further substitution
with the F atom at para-phenyl position enhanced cytotox-
icities against both Jurkat and H226 cancer cells, while sub-
stitution with a Cl or a Br atom at the same position led to
the inactive compounds of 3c and 3d. Substitution with an
electron-donating group such as OMe, phenyl, or naphthyl
moiety at the para-phenyl position did not significantly
affect the inhibitory activity of Jurkat cancer cells in which
compounds 3e, 3f, and 3g exhibited IC50 in the range
6.08–9.28 μM. Quinazolinone 4 was weakly active against
both Jurkat and H226 cancer cells, indicating that oxime is
more favorable than its ketone precursor 2. However, its
methyloxime derivative 5 become inactive, indicating that a
hydrogen-bond-donating group is required. The same trend
was observed in most of the oxime derivatives 6a, 6b, and
6e–6g, which were active against Jurkat cancer cells, while
none of their methyloxime counterparts 7a–7g exhibited
cytotoxicity at 20 μM. Among these oxime derivatives 6a–
6g, the electron-withdrawing groups such as Cl and Br were
The preparation of oxime, methyloxime, and amide-
containing quinazolinone derivatives is illustrated in
Scheme 1. Reaction of 4-hydroxyquinazoline (1) with
chloroacetone under basic conditions gave exclusively qui-
nazolinone 2 in 88% yield. Alkylation of 1 occurred at N-3
but not at N-1 or the carbonyl oxygen was confirmed by
NMR spectra. Structure of 2 was further proved by HMBC
in which the coupling of H-2 (8.21 ppm) with C-2
(148 ppm, J-1), C-4 (148 ppm, J-3), and C-1' (55 ppm, J-3),
respectively, was observed (Figure 2). Treatment of 1 with
phenacyl bromide under basic conditions gave exclusively
O
O
H
N
O
N
O
H
H
O
N
O
I
O
O
IV
H
O
N
N
H
O
O
N
N
Ar
O
F
O
II
Target compounds
H
N
O
N
O
H
Ar = biphenyl, naphthalene, etc
O
III
FIGURE 1 Structures of compounds I–IV and the target compounds

CHANG ET AL.
3
OH
N
O
N
O
N
NH2OR • HCl
EtOH
Ar
Ar
N
N
BrCH2COAr
K2CO3, DMF
N
O
N
70-88%
OR
87-95%
1
6 a-g R = H
7 a-g R = Me
3 a-g
BrCH2COMe
NaN3
K2CO3, DMF
88%
H2SO4, 0 °C
84-93 %
O
N
N
O
N
H
N
Me
N
Ar
O
O
2
8 a-g
NH2OR • HCl
EtOH
a
: Ar = Ph
b: Ar = Ph-4-F
c: Ar = Ph-4-Cl
d: Ar = Ph-4-Br
75-77%
e
: Ar = Ph-4-OMe
O
f: Ar = Ph-4-phenyl
g: Ar = 2-Naphthyl
Me
OR
N
N
N
SCHEME 1 Synthesis of quinazolinone
derivatives
4
5
R = H
R = Me
unfavorable, so compounds 6c and 6d were inactive.
However, compounds 6e, 6f, and 6g, which bear electron-
donating groups such as OMe, phenyl, and naphthyl,
improved the antiproliferative activities against both Jurkat
and H226 cancer cells. The same structure–activity relation-
ship (SAR) was obtained for the amide-containing quinazo-
linone derivatives 8a–8g, in which compounds 8c and 8d
bearing the electron-withdrawing groups such as Cl and Br
were inactive. Compounds 8a, 8b, and 8e were weakly
active against Jurkat cancer cells, while compounds 8f and
8g, which bear the electron-donating groups phenyl and
naphthyl, respectively, were active against all three cancer
cells tested. Our results indicate that quinazolinone 8g and
its biphenyl counterpart 8f show more potent antiprolifera-
tive activities than the positive control methotrexate against
all three cancer cell lines tested (Table 1). Among them,
compound 8g was the most active against NPC-TW01 with
an IC50 value of 4.78 μM and therefore was subjected for
further study on NPC-TW01 cell cycle distribution.
Figure 3 shows that compound 8g induced NPC-TW01 cell
arrest at the G1/G0 phase in a time- and concentration-
dependent manner. After 48 hr of treatment, significant
G1/G0 (~70%) cell accumulation and a corresponding
decrease in the G2/M and S phases cell population were
detected. Moreover, a characteristic hypo-diploid DNA con-
tent peak (sub-G1) was found to increase from 1 to 3.6%
and 4.4% in NPC-TW01 cells treated with 8g for 48 and
72 hr (Table 2). These results indicate that 8g can induce
cells arrest in the G1/G0 phase and cause cell death. Further
structural optimization of 8g and detailed study of its anti-
proliferative mechanism are going on.
3
|
EXPERIMENTAL
3.1 | General
Thin-layer chromatography (TLC) was carried out on
precoated (0.2 mm) silica gel 60 F254 plates from EM
Laboratories, Inc.; detection was by UV light (254 nm).
Melting points were measured on an Electrothermal IA9100
digital melting point apparatus; the values are uncorrected.
1H-NMR spectra were recorded on a Varian Unity-400 spec-
trometer at 400 MHz or Varian Gemini-200 spectrometer at
200 MHz. The chemical shifts δ are reported in ppm with
SiMe4 as the internal standard (= 0 ppm); the coupling
148
O
4.96
55
2.25
H
H
202
160
CH3
27
4
5
8
N
6
7
3
2
O
1
N
H
148
8.21
4.96
FIGURE 2 HMBC spectrum of 3-(2-oxopropyl)quinazolin-4(3H)-one (2)

4
CHANG ET AL.
TABLE 1 In vitro antiproliferative activity of quinazolinone bearing
ketone, oxime, methyloxime, and amide derivatives
(a)
80
60
40
20
0
IC50 (μM)a
Compound
R
TW01
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
19.50
Jurkat
>20.00
8.79
H226
G1/G0
S
G2/M
Debris
2
Me
>20.00
>20.00
13.8
3a
Ph
3b
Ph-4-F
8.22
3c
Ph-4-Cl
Ph-4-Br
Ph-4-OMe
Ph-4-phenyl
Ph-4-naphthyl
Me
>20.00
>20.00
9.28
>20.00
>20.00
>20.00
>20.00
>20.00
12.27
3d
3e
3f
7.64
3g
6.08
4
6.55
0
24
48
72
96
5
Me
>20.00
7.16
>20.00
>20.00
>20.00
>20.00
>20.00
14.87
Treatment period (Hour)
6a
Ph
(b) 80
6b
Ph-4-F
7.69
6c
Ph-4-Cl
Ph-4-Br
Ph-4-OMe
Ph-4-phenyl
Ph-4-naphthyl
Ph
>20.00
>20.00
6.59
6d
G1/G0
S
G2/M
Debris
60
40
20
0
6e
6f
6.41
10.41
6g
6.20
7.10
7a
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
7.80
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
7.63
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
>20.00
5.84
7b
Ph-4-F
7c
Ph-4-Cl
Ph-4-Br
Ph-4-OMe
Ph-4-phenyl
Ph-4-naphthyl
Ph
7d
7e
7f
7g
0
5
10
15
20
25
30
35
40
45
8a
Compound Concentration uM
8b
Ph-4-F
7.05
FIGURE 3 Compound 8g inducing concentration- and time-dependent
G1/G0 cell arrest in human NPC-TW01 cell line. NPC-TW01 cells were
treated with 1 × IC50 (4.78 μM) of 8g for 24, 48, and 72 hr (a) or different
concentrations (2.5, 5, 10, 20, and 40 μM) for 48 hr (b). The cell population
ratios were determined by flow cytometry
8c
Ph-4-Cl
Ph-4-Br
Ph-4-OMe
Ph-4-phenyl
Ph-4-naphthyl
>20.00
>20.00
6.69
8d
8e
8f
11.22
8.52
8g
4.78
7.88
The yellow solid thus obtained was collected and purified by
column chromatography on silica gel using n-hexane/ethyl
acetate 1:2. The proper fractions were combined and evapo-
rated to furnish a residual solid which was crystallized from
Et2O and CH2Cl2.
Methotrexate
43.4
18.70
22.93
a
Cancer cells were treated with different quinazolinone compounds of various
concentrations for 72 hr, and cell survival was determined using MTT colori-
metric assay. Representative data from three independent experiments per-
formed in quadruplicate are shown.
3-(2-Oxopropyl)quinazolin-4(3H)-one (2): Yield 88%.
constants J are in Hz. Elemental analyses were carried out
on a Heraeus CHN-O-Rapid elemental analyzer, and results
were within Æ0.4% of the calculated values.
1
ꢀ
mp 170–171 C. H-NMR (DMSO-d6) 2.25 (s, 3H, Me),
4.96 (s, 2H, NCH2), 7.56 (dd, J = 0.8, 8.0 Hz, 1H-C (8)),
7.71 (d, J = 8.0 Hz, 1H-C (7)), 7.85 (dd, J = 1.6, 8.0 Hz,
1H-C (6)), 8.15 (dd, J = 1.6, 8.0 Hz, 1H-C (5)), 8.21
(s, 1H-C (2)). 13C-NMR (DMSO) 27.19 (Me), 54.72
(CH2CO), 121.41, 126.08, 127.24, 127.29, 134.59, 148.01,
3.2 | Syntheses
3.2.1
| Preparation of 2 and 3a-g
General procedure: 4-Hydroxyquinazoline (1) (10 mmol),
K2CO3 (10 mmol), and dry DMF (50 mL) were stirred at
room temperature (r.t.) for 30 min. To this solution was
added chloroacetone/2-(bromoacetyl)naphthalene/substituted
phenacyl bromide (10 mmol) in dry DMF (10 mL) in one
portion. The resulting mixture was stirred at r.t. for 24 hr
(TLC monitoring) and then poured into ice-water (100 mL).
TABLE 2 Compound 8g arrested NPC-TW01 cells at G0/G1 phase
Cell distribution (%)
Control
49.73
26.61
21.82
1.84
24 hr
46.04
26.32
24.74
2.90
48 hr
70.43
16.94
8.97
72 hr
71.97
15.63
8.00
G0/G1
G2/M
S
Debris
3.66
4.40

CHANG ET AL.
5
148.1, 160.02 (C (4)), 201.73 (CH2CO). Anal. Calcd for
C11H10N2O2: C 65.34, H 4.98, N 13.85. Found: C 65.21, H
4.96, N 13.86.
160.30 (C (4)), 163.98 (C-OCH3), 191.17 (CH2CO). Anal.
Calcd for C17H14N2O3: C 69.38, H 4.79, N 9.52. Found: C
69.38, H 4.81, N 9.56.
3-(2-Oxo-2-phenylethyl)quinazolin-4(3H)-one (3a): Yield
3-(2-[Biphenyl-4-yl]-2-oxoethyl)quinazolin-4(3H)-one
90%. mp 228–229 C. 1H-NMR (DMSO-d6) 5.64 (s, 2H,
(3f): Yield 87%. mp 228–229 C. 1H-NMR (DMSO-d6)
ꢀ
ꢀ
NCH2), 7.57–7.64 (m, 3H, arom. H), 7.73–7.70 (m, 2H, arom.
H), 7.90 (dd, J = 1.6, 8.0 Hz, 1H, arom. H), 8.11 (dd, J = 1.2,
8.0 Hz, 2H, arom. H), 8.17 (dd, J = 1.2, 8.0 Hz, 1H-C (5)),
8.35 (s, 1H-C (2)). 13C-NMR (DMSO) 52.32 (CH2CO),
121.51, 126.21, 127.45, 127.46, 128.24, 129.22, 134.38,
134.42, 134.81, 148.17, 148.44, 160.33 (C (4)), 192.96
(CH2CO). Anal. Calcd for C16H12N2O2: C 72.72, H 4.58, N
10.60. Found: C 72.74, H 4.54, N 10.60.
3-(2-[4-Fluorophenyl]-2-oxoeꢀthyl)quinazolin-4(3H)-one
(3b): Yield 94%. mp 228–229 C. 1H-NMR (DMSO-d6)
5.63 (s, 2H, NCH2), 7.48 (dd, J = 2.4, 7.2 Hz, 2H, arom.
H), 7.61 (d, J = 1.2, 7.2 Hz, 1H-C (6)), 7.75 (d, J = 7.6 Hz,
1H-C (8)), 7.90 (dd, J = 1.2, 7.2 Hz, 1H-C (7)), 8.14–8.21
(m, 3H, arom. H), 8.34 (s, 1H-C (2)). 13C-NMR (DMSO)
52.17 (CH2CO), 116.15, 116.37, 121.45, 126.14, 127.40,
131.11, 131.14, 131.27, 131.37, 134.73, 148.12, 148.33,
160.22 (C (4)), 164.38, 166.90, 191.56 (CH2CO). Anal.
Calcd for C16H11FN2O2Á0.2 H2O: C 67.21, H 4.03, N 9.80.
Found: C 66.85, H 4.02, N 9.73.
5.67 (s, 2H, NCH2), 7.46–7.62 (m, 4H, arom. H), 7.74–7.81
(m, 3H, arom. H), 7.87–7.94 (m, 3H, arom. H), 8.16
(d, J = 1.2 Hz, 1H, arom. H), 8.18 (s, 1H, arom. H), 8.20
(d, J = 2.0 Hz, 1H, arom. H), 8.37 (s, 1H-C (2)). 13C-NMR
(DMSO) 52.29 (CH2CO), 121.50, 126.18, 127.20, 127.28,
127.43, 128.77, 128.97, 129.30, 133.17, 134.77, 138.75,
145.58, 148.17, 148.43, 160.29 (C(4)), 192.48 (CH2CO).
Anal. Calcd for C22H16N2O2: C 77.63, H 4.74, N 8.23.
Found: C 77.48, H 4.67, N 8.16.
3-(2-[Naphthalen-2-yl]-2-oxoethyl)quinazolin-4(3H)-one
(3g): Yield 90%. mp 228–229 C. 1H-NMR (DMSO-d6)
ꢀ
5.76 (s, 2H, NCH2), 7.59 (dd, J = 1.2, 8.0 Hz, 1H-C (6)),
7.63–7.74 (m, 3H, arom. H), 7.89 (dd, J = 1.6, 8.4 Hz, 1H,
arom. H), 8.01–8.09 (m, 3H, arom. H), 8.18 (m, 2H, arom.
H), 8.38 (s, 1H, arom. H), 8.87 (s, 1H-C (2)). 13C-NMR
(DMSO) 52.29 (CH2CO), 121.51, 123.36, 126.19, 127.40,
127.42, 127.94, 128.83, 129.30, 129.81, 130.48, 131.67,
132.22, 134.77, 135.54, 148.17, 148.46, 160.32 (C (4)),
192.86 (CH2CO). Anal. Calcd for C20H14N2O2: C 76.42, H
4.49, N 8.91. Found: C 76.12, H 4.48, N 8.87.
3-(2-[4-Chlorophenyl]-2-oxoethyl)quinazolin-4(3H)-one
(3c): Yield 92%. mp 219–220 C. 1H-NMR (DMSO-d6)
ꢀ
5.63 (s, 2H, NCH2), 7.59 (dd, J = 1.2, 7.8 Hz, 1H-C (8)),
7.69–7.75 (m, 3H, arom. H), 7.85 (dd, J = 1.6, 8.0 Hz,
1H-C (7)), 7.91 (m, 1H, arom. H), 8.10–8.16 (m,3H, arom.
H), 8.34 (s, 1H-C (2)). 13C-NMR (DMSO): 52.16 (CH2CO),
121.39, 126.09, 127.33, 127.35, 129.24, 130.06, 133.0,
134.68, 139.16, 148.08, 148.24, 160.15 (C (4)), 192.0
(CH2CO). Anal. Calcd for C16H11ClN2O2: C 64.33, H 3.71,
N 9.38. Found: C 64.32, H 3.88, N 9.26.
3.2.2
| Preparation of 4 and 6a–g
General procedure: A solution of 2/3a–g (1 mmol) was
added a solution of hydroxylamine hydrochloride (2 mmol)
in EtOH (20 mL). The mixture was heated at reflux for 4 hr
(TLC monitoring) and evaporated to give a residual solid.
The white solid thus obtained was collected, purified by
flash column chromatography (FC; silica gel; CH2Cl2/
MeOH 15:1), and recrystallized from Et2O and CH2Cl2.
(E)-3-(2-[Hydroxyimino]propyl)quinazolin-4(3H)-one
(4): Yield 75%. mp 170–171 C. H-NMR (DMSO-d6) 1.82
(s, 3H, Me), 4.74 (s, 2H, NCH2), 7.58 (dd, J = 1.2, 8.0 Hz,
1H-C (6)), 7.69 (d, J = 8.4 Hz, 1H-C (8)), 7.86 (dd,
J = 1.6, 8.4 Hz, 1H-C (7)), 8.17 (dd, J = 1.6, 8.0 Hz, 1H-C
(5)), 8.30 (s, 1H-C (2)), 10.80 (s, NOH). 13C-NMR (DMSO)
12.36 (Me), 48.28 (NCH2), 121.64, 126.11, 127.11, 127.14,
134.43, 147.50, 148.35, 150.74 (C=NOH), 159.99 (C=O).
Anal. Calcd for C11H11N3O2: C 60.82, H 5.10, N 19.34.
Found: C 60.86, H 5.10, N 19.33.
3-(2-[4-Bromophenyl]-2-oxoethyl)quinazolin-4(3H)-one
(3d): Yield 95%. mp 211–212 C. 1H-NMR (DMSO-d6)
ꢀ
1
ꢀ
5.68 (s, 2H, NCH2), 7.59 (dd, J = 1.2, 7.8 Hz, 1H-C (8)),
7.82 (dd. J = 0.8, 8.0 Hz, 1H-C (7)), 7.97 (m, 3H, arom. H),
7.73–7.70 (m, 2H, arom. H), 7.90 (m, 1H, arom. H), 8.11
(dd, J = 1.2, 8.0 Hz, 2H, arom. H), 8.23 (dd, J = 1.2,
8.0 Hz, 1H-C (5)), 8.39 (s, 1H-C (2)). 13C-NMR (DMSO)
52.30 (CH2CO), 121.48, 126.24, 127.47, 127.55, 128.56,
130.27, 132.34, 133.42, 134.90, 148.16, 148.38, 160.34
(C (4)), 192.36 (CH2CO). Anal. Calcd for C16H11BrN2O2: C
56.00, H 3.23, N 8.16. Found: C 55.91, H 3.25, N 8.15.
3-(2-[4-Methoxyphenyl]-2-oxoethyl)quinazolin-4(3H)-
(Z)-3-(2-[Hydroxyimino]-2-phenylethyl)quinazolin-4
(3H)-one (6a): Yield 88%. mp 228–229 C. 1H-NMR
ꢀ
1
ꢀ
one (3e): Yield 95%. mp 228–229 C. H-NMR (DMSO-d6)
3.88 (s, 3H, OCH3), 5.58 (s, 2H, NCH2), 7.14 (dd, J = 2,
6.8 Hz, 2H, arom. H), 7.60 (d, J = 1.2, 8 Hz, 1H-C (6)),
7.75 (d, J = 8.0 Hz, 1H-C (8)), 7.88 (dd, J = 1.2, 7.2 Hz,
1H-C (7)), 8.09 (dd, J = 2.0, 6.8 Hz, 2H, arom. H), 8.16
(dd, 3H, arom. H), 8.34 (s, 1H-C (2)). 13C-NMR (DMSO)
51.89 (CH2CO), 55.79 (OCH3), 114.37, 121.51, 126.17,
127.27, 127.37, 127.40, 130.61, 134.71, 148.17, 148.50,
(DMSO-d6) 5.24 (s, 2H, NCH2), 7.29–7.36 (m, 3H, arom.
H), 7.53 (d, J = 1.2, 8.4 Hz, 1H-C (6)), 7.62–7.65 (m, 3H,
arom. H), 7.82 (dd, J = 1.2, 8.4 Hz, 1H-C (7)), 8.08
(dd, J = 1.2, 8.0 MHz, 1H-C (5)), 8.43 (s, 1H-C (2)), 11.80
(s, NOH). 13C-NMR (DMSO) 42.19 (NCH2), 121.25,
125.96, 126.73, 127.11, 128.23, 128.85, 134.40, 134.51,
147.62, 148.48, 152.64 (C=O), 159.98 (C=NOH).

6
CHANG ET AL.
Anal. Calcd for C16H13N3O2: C 68.81, H 4.69, N 15.05.
Found: C 68.82, H 4.69, N 14.94.
128.96, 133.69, 1,349.44, 139.28, 140.42, 147.64, 148.55,
152.13 (C=O), 160.05 (C=NOH). Anal. Calcd for
C22H17N3O2: C 74.35, H 4.82, N 11.82. Found: C 74.17, H
4.82, N 11.81.
(Z)-3-(2-(4-Fluorophenyl)-2-(hydroxyimino)ethyl)quina-
1
ꢀ
zolin-4(3H)-one (6b): Yield 77%. mp 228–229 C. H-NMR
(DMSO-d6) 5.22 (s, 2H, NCH2), 7.16–7.21 (m, 2H, arom.
H), 7.57 (d, J = 1.2, 8.0 Hz, 1H-C (6)), 7.65 (dd, J = 0.8,
8.2 Hz, arom. H), 7.70–7.83 (m, 3H, arom. H), 8.09
(dd, J = 1.6, 8.0 Hz, 1H-C (5)), 8.46 (s, 1H-C (2)), 11.83
(s, NOH). 13C-NMR (DMSO) 42.46 (NCH2), 115.07,
115.29, 121.23, 125.97, 127.12, 127.14, 128.95, 129.03,
131.1, 131.13, 134.44, 147.63, 148.60, 151.82, 160.03
(C=O), 161.19 (C=NOH), 163.63. Anal. Calcd for
C16H12FN3O2: C 64.64, H 4.07, N 14.13. Found: C 64.58,
H 4.08, N 14.13.
(Z)-3-(2-(Hydroxyimino)-2-(naphthalen-2-yl)ethyl)quina-
1
ꢀ
zolin-4(3H)-one (6g): Yield 80%. mp 228–229 C. H-NMR
(DMSO-d6) 5.34 (s, 2H, NCH2), 7.46–7.52 (m, 3H, arom.
H), 7.62 (dd, J = 0.4, 8.4 Hz, 1H, arom. H), 7.75–7.93
(m, 5H, arom. H), 8.07 (dd, J = 1.6, 8.0 Hz, 1H-C (5)), 8.24
(d, J = 1.2 Hz, arom. H), 8.49 (s, 1H-C (2)), 11.94
(s, NOH). 13C-NMR (DMSO) 42.25 (NCH2), 121.31,
124.14, 126.02, 126.31, 126.6, 126.8, 127.15, 127.23,
127.58, 127.81, 128.34, 132.13, 132.6, 132.94, 134.51,
147.66, 148.66, 152.48 (C=O), 160.14 (C=NOH). Anal.
Calcd for C20H15N3O2: C 72.94, H 4.59, N 12.76. Found: C
72.56, H 4.54, N 12.67.
(Z)-3-(2-(4-Chlorophenyl)-2-(hydroxyimino)ethyl)quina-
ꢀ
zolin-4(3H)-one (6c): Yield 76%. mp 210–211 C.
11H-NMR (DMSO-d6) 5.19 (s, 2H, NCH2), 7.39–7.43 (m,
2H, arom. H), 7.53 (dd, J = 1.2, 8.0 Hz, 1H-C (8)),
7.63–7.70 (m, 3H, arom. H), 7.82 (m, 1H, arom. H), 8.07
(dd, J = 1.6, 8.0 Hz, 1H-C (5)), 8.44 (s, 1H-C (2)), 11.94
(s, NOH). 13C-NMR (DMSO) 42.68 (NCH2), 121.32,
126.12, 127.26, 127.4, 128.47, 128.69, 133.72, 133.78,
134.71, 147.73, 148.84, 151.9 (C=NOH), 160.24 (C=O).
Anal. Calcd for C16H12ClN3O2: C 61.25, H 3.86, N 13.39.
Found: C 61.20, H 3.82, N 13.37.
3.2.3
| Preparation of 5 and 7a–g
General procedure: A solution of 2/5a–g (1 mmol) was
added a solution of O-methylhydroxylamine hydrochloride
(2 mmol) in EtOH (20 mL). The mixture was heated at
reflux for 4 h (TLC monitoring) and evaporated to give a
residual solid. The white solid thus obtained was collected,
purified by flash column chromatography (FC; silica gel; n-
hexane/EtOAc 1:3), and recrystallized from Et2O and
CH2Cl2.
(Z)-3-(2-(4-Bromophenyl)-2-(hydroxyimino)ethyl)quina-
1
ꢀ
(E)-3-(2-[Methoxyimino]propyl)quinazolin-4(3H)-one
zolin-4(3H)-one (6d): Yield 75%. mp 194–195 C. H-NMR
(DMSO-d6) 5.24 (s, 2H, NCH2), 7.56–7.3662 (m, 3H, arom.
H), 7.66–7.71 (m, 3H, arom. H), 7.89 (dd, J = 1.2, 8.4 Hz,
1H-C (7)), 8.13 (dd, J = 1.2, 8.0 MHz, 1H-C (5)), 8.50
(s, 1H-C (2)), 12.04 (s, NOH). 13C-NMR (DMSO) 42.73
(NCH2), 121.37, 122.57, 126.20, 127.31, 127.52, 129.02,
131.47, 134.16, 134.82, 147.76, 148.90, 152.07 (C=O),
160.34 (C=NOH). Anal. Calcd for C16H12BrN3O2: C 53.65,
H 3.38, N 11.73. Found: C 53.73, H 3.41, N 11.60.
(5): Yield 77%. mp 69–70 C. 1H-NMR (DMSO-d6) δ:
ꢀ
1.81 (s, 3H, CH3), 3.64 (s, 3H, NOCH3), 4.73 (s, 2H,
NCH2), 7.58 (ddd, J = 1.6, 8.0, 1.2 Hz, 1H-C (6)), 7.70
(dd, J = 0.8, 8.0 Hz, 1H-[C7]), 7.86 (ddd, J = 1.2, 8.0,
0.8 Hz, 1H-C (8)), 8.16 (dd, 1H, J = 1.2, 8.0 Hz, 1H-C
(8)), 8.28 (s, 1H-C (2)). 13C-NMR (DMSO) 13.02 (CH3),
48.27 (NOCH3), 61.43 (NCH2), 121.61, 126.33, 127.34,
134.7, 148.0, 148.27, 152.71 (C=O), 160.17
(C=NOCH3). Anal. Calcd for C12H13N3O2: C 62.33, H
5.67, N 18.17. Found: C 62.37, H 5.70, N 18.08.
(Z)-3-(2-[Methoxyimino]-2-phenylethylꢀ)quinazolin-4
(3H)-one (7a): Yield 70%. mp 97–98 C. 1H-NMR
(DMSO-d6) 3.91 (s, 3H, NOCH3), 5.22 (s, 2H, NCH2),
7.31–7.33 (m, 3H, arom. H), 7.47–7.62 (m, 4H, arom.
H), 7.80 (ddd, J = 1.6, 8.0, 2.0 Hz, 1H-C (8)), 8.04 (dd,
J = 1.2, 8.0 Hz, 1H-C (8)), 8.41 (s, 1H-C (2)). 13C-
NMR (DMSO) 43.34 (NOCH3), 62.41 (NCH2), 121.27,
126.13, 127.28, 127.47, 128.47, 129.53, 133.52, 134.75,
147.71, 148.58, 154.31 (C=O), 160.22 (C=NOCH3).
Anal. Calcd for C17H15N3O2: C 69.61, H 5.15, N 14.33.
Found: C 69.71, H 5.18, N 14.33.
(Z)-3-(2-(Hydroxyimino)-2-(4-methoxyphenyl)ethꢀyl)qui-
nazolin-4(3H)-one (6e): Yield 77%. mp 228–229 C. 1H-
NMR (DMSO-d6) 3.73 (s, 3H, OCH3), 5.21 (s, 2H, NCH2),
6.92 (dd, J = 2.0, 7.2, 2H, arom. H), 7.54 (d, J = 1.2,
8.0 Hz, 1H-C (6)), 7.60–7.65 (m, 3H, arom. H), 7.83 (dd,
J = 1.6, 8.0 Hz, 1H-C (7)), 8.10 (dd, J = 1.2, 8.0 Hz, 1H-C
(5)), 8.40 (s, 1H-C (2)), 11.64 (s, NOH). 13C-NMR (DMSO)
41.77 (NCH2), 55.12 (OCH3), 113.7, 121.3, 126.01, 126.8,
127.12, 128.08, 134.41, 147.61, 148.4, 152.11 (C=O),
159.79 (C-OCH3), 159.99 (C=NOH). Anal. Calcd for
C17H15N3O3: C 66.01, H 4.89, N 13.58. Found: C 66.01, H
4.92, N 13.60.
(Z)-3-(2-(Biphenyl-4-yl)-2-(hydroxyimino)ethyl)quinazo-
1
ꢀ
lin-4(3H)-one (6f): Yield 75%. mp 228–229 C. H-NMR
(DMSO-d6) 5.27 (s, 2H, NCH2), 7.34–7.54 (m, 4H, arom.
H), 7.64–7.69 (m, 5H, arom. H), 7.78–7.83 (m, 3H, arom.
H), 8.11 (dd, J = 1.2, 8.0 Hz, 1H-C (5)), 8.47 (s, 1H–C (2)),
11.88 (s, NOH). 13C-NMR (DMSO) 42.03 (NCH2), 121.3,
125.0, 126.48, 126.59, 127.12, 127.14, 127.19, 127.71,
(Z)-3-(2-(4-Fluorophenyl)-2-(methoxyimino)ethyl)quina-
1
ꢀ
zolin-4(3H)-one (7b): Yield 71%. mp 102–103 C. H-NMR
(DMSO-d6) 3.90 (s, 3H, NOCH3), 5.19 (s, 2H, NCH2),
7.13–7.19 (m, 2H, arom. H), 7.52 (dd, J = 1.2, 8.0 Hz, 1H-
C (6)), 7.62–7.67 (m, 3H, arom. H), 7.81 (dd, J = 1.6,
8.0 Hz, 1H-C (7)), 8.05 (dd, J = 1.6, 8.0 Hz, 1H-C (8)),

CHANG ET AL.
7
8.42 (s, 1H-C (2)). 13C-NMR (DMSO) 43.5 (NOCH3),
62.45 (NCH2), 115.35, 115.60, 121.24, 126.12, 127.32,
127.49, 129.57, 129.65, 130.12, 130.15, 134.78, 147.72,
148.61, 153.41 (C=O), 160.25, 161.58, 164.03
(C=NOCH3). Anal. Calcd for C17H14FN3O2: C 65.59, H
4.53, N 13.50. Found: C 65.57, H 4.55, N 13.49.
(Z)-3-(2-(Methoxyimino)-2-(naphthalen-2-yl)ethyl)quina-
1
ꢀ
zolin-4(3H)-one (7g): Yield 78%. mp 173–174 C. H-NMR
(DMSO-d6) 3.99 (s, 3H, NOCH3), 5.36 (s, 2H, NCH2),
7.47–7.55 (m, 3H, arom. H), 7.63 (dd, J = 0.4, 8.0, 1H-C
(6)), 7.75–7.80 (m, 2H, arom. H), 7.87–7.94 (m, 3H, arom.
H), 8.06 (dd, J = 1.2, 8.0 Hz, 1H-C (8)), 8.23 (d, J = 1.2 Hz,
1H, arom. H), 8.52 (s, 1H-C (2)). 13C-NMR (DMSO) 43.08
(NOCH3), 62.44 (NCH2), 121.21, 124.28, 126.0, 126.72,
126.86, 127.06, 127.20, 127.30, 127.61, 127.89, 128.38,
130.99, 132.49, 133.08, 134.59, 147.63, 148.52, 153.86
(Z)-3-(2-(4-Chlorophenyl)-2-(methoxyimino)ethyl)quina-
ꢀ
zolin-4(3H)-one (7c): Yield: 80%. mp 145–146 C.
1H-NMR (DMSO-d6) 3.91 (s, 3H, NOCH3), 5.19 (s, 2H,
NCH2), 7.42 (dd, J = 2.8, 8.4 Hz, 2H, arom. H), 7.52 (ddd,
J = 1.2, 8.0, 1.2 Hz, 1H-C (8)), 7.65 (dd, J = 2.8, 8.4 Hz,
3H, arom. H), 7.82 (m, 1H, arom. H), 8.16 (dd, 1H, J = 1.2,
8.0 Hz, 1H-C (5)), 8.43 (s, 1H-C (2)). 13C-NMR (DMSO)
43.40 (NOCH3), 62.57 (NCH2), 121.23, 126.12, 127.33,
127.51, 128.56, 129.07, 132.59, 134.3, 134.81, 147.72,
148.63, 153.29 (C=O), 160.26 (C=NOCH3). Anal. Calcd
for C17H14ClN3O2: C 62.30, H 4.31, N 12.82. Found: C
62.17, H 4.31, N 12.78.
(C=O),
160.15
(C=NOCH3).
Anal.
Calcd
for
C20H15N3O2: C 72.94, H 4.59, N 12.76. Found: C 72.56,
H 4.54, N 12.67.
3.2.4
| Preparation of 8a–g
General procedure: A solution of 3a–g (1 mmol) in H2SO4
(3 mL) was stirred at r.t. for 10 min. To this solution, was
added sodium azide (2 mmol) in one portion. The mixture
was stirred continuously at r.t. for 1 hr (TLC monitoring)
and then poured into ice-water (100 mL). The white solid
thus obtained was collected and purified by flash column
chromatography (FC; silica gel; CH2Cl2/MeOH 30:1). The
proper fractions were combined and evaporated to furnish a
residual solid which was crystallized from Et2O and
CH2Cl2.
(Z)-3-(2-(4-Bromophenyl)-2-(methoxyimino)ethyl)quina-
ꢀ
zolin-4(3H)-one (7d): Yield: 83%. Mp 142–143 C.
1H-NMR (DMSO-d6) 3.91 (s, 3H, NOCH3), 5.18 (s, 2H,
NCH2), 7.45–7.58 (m, 5H, arom. H), 7.64 (d, J = 8.0 Hz,
1H-C (8)), 7.82 (m, 1H, arom. H), 8.04 (dd, 1H, J = 1.6,
8.0 Hz, 1H-C (5)), 8.42 (s, 1H-C (2)). 13C-NMR (DMSO)
43.34 (NOCH3), 62.58 (NCH2), 121.23, 123.05, 126.12,
127.33, 127.52, 129.31, 131.48, 132.97, 134.82, 147.72,
148.63, 153.36 (C=O), 160.26 (C=NOCH3). Anal. Calcd
for C17H14BrN3O2: C 54.86, H 3.79, N 11.29. Found: C
54.89, H 3.85,N 11.31.
2-(4-Oxoquinazolin-3[4H]-yl)-N-phenylacetamide (8a):
Yield 90%. mp 252–253 C. 1H-NMR (DMSO-d6) 4.83
ꢀ
(s, 2H, NCH2), 7.29 (s, 1H, arom. H), 7.33 (d, J = 7.6, 2H,
arom. H), 7.53–7.57 (m, 3H, arom. H), 7.72 (d, J = 8.0 Hz,
1H, arom. H), 7.83–7.85 (m, 1H, arom. H), 8.12–8.15
(m, 1H, arom. H), 8.33 (s, 1H-C (2)), 10.51 (s, 1H, CONH).
13C-NMR (DMSO) 49.34 (NCH2), 119.68, 121.73, 124.35,
126.52, 127.63, 127.91, 129.46, 135.29, 138.84, 148.33,
149.02, 160.94 (C=O), 165.94 (CONH). Anal. Calcd for
C16H13N3O2: C 68.81, H 4.69, N 15.05. Found: C 68.66, H
4.54, N 15.01.
N-(4-Fluorophenyl)-2-(4-oxoquinazolin-3[4H]-yl)acetamide
(8b): Yield 91%. mp 244–245 ꢀC. 1H-NMR (DMSO-d6)
4.85 (s, 2H, NCH2), 7.14–7.19 (m, 2H, arom. H), 7.55–7.62
(m, 3H, arom. H), 7.73 (d, J = 7.6 Hz, 1H, arom. H),
7.84–7.88 (m, 1H, arom. H), 8.16 (dd, J = 1.2, 8.0 Hz, 1H-
C (8)), 8.37 (s, 1H-C (2)), 10.54 (s, 1H, CONH). 13C-NMR
(DMSO) 48.84 (NCH2), 115.48, 115.70, 120.9, 120.98,
121.49, 126.11, 127.30, 127.34, 134.68, 135.06, 135.08,
148.15, 148.66, 157.03, 159.42, 160.41 (C=O), 165.47
(C=NOCH3). Anal. Calcd for C16H12FN3O2: C 64.64, H
4.07, N 14.13. Found: C 64.57, H 4.06, N 14.13.
(Z)-3-(2-(Methoxyimino)-2-(4-methoxyphenyl)ethyl)qui-
ꢀ
nazolin-4(3H)-one (7e): Yield: 75%. mp 113–114 C.
1H-NMR (DMSO-d6) 3.70 (s, 3H, OCH3), 3.89 (s, 3H,
NOCH3), 5.18 (s, 2H, NCH2), 6.89 (dd, J = 2.4, 9.2 Hz,
2H, arom. H), 7.48–7.57 (m, 3H, arom. H), 7.63
(dd, J = 0.8, 8.0, Hz, 1H-C (6)), 7.81 (ddd, J = 1.6, 8.0,
1.2 Hz, 1H-C (7)), 8.06 (dd, J = 1.6, 8.0 Hz, 1H-C (8)),
8.39 (s, 1H-C (2)). 13C-NMR (DMSO) 42.98 (NOCH3),
55.33 (OCH3), 62.24 (NCH2), 113.9, 121.32, 125.80,
126.17, 127.30, 127.46, 128.71, 134.75, 147.71, 148.54,
153.69 (C=O), 160.21, 160.26 (C=NOCH3). Anal. Calcd
for C18H17N3O3: C 66.86, H 5.30, N 13.00. Found C 66.68,
H 5.37, N 13.04.
(Z)-3-(2-(Biphenyl-4-yl)-2-(methoxyimino)ethyl)quinazo-
1
ꢀ
lin-4(3H)-one (7f): Yield: 71%. mp 146–147 C. H-NMR
(DMSO-d6) 3.93 (s, 3H, NOCH3), 5.24 (s, 2H, NCH2),
7.32–7.52 (m, 4H, arom. H), 7.62–7.66 (m, 5H, arom. H),
7.71–7.81 (m, 3H, arom. H), 8.07 (dd, J = 1.2, 8.0 Hz, 1H-
C (8)), 8.45 (s, 1H-C (2)). 13C-NMR (DMSO) 43.17
(NOCH3), 62.49 (NCH2), 121.32, 126.17, 126.66, 126.79,
127.32, 127.48, 127.80, 128.05, 129.20, 132.71, 134.78,
139.25, 141.04, 147.74, 148.64, 153.69 (C=O), 160.28
(C=NOCH3). Anal. Calcd for C23H19N3O2: C 74.78, H
5.18, N 11.37. Found: C 74.62, H 5.16, N 11.29.
N-(4-Chlorophenyl)-2-(4-oxoquinazolin-3[4H]-yl)acetamide
(8c): Yield 89%. mp 262–263 ꢀC. 1H-NMR (DMSO-d6)
4.84 (s, 2H, NCH2), 7.39 (dd, J = 2.8, 9.2 Hz, 2H, arom.
H), 7.55–7.60 (m, 3H, arom. H), 7.72 (d, J = 8.0 Hz, 1H-C
(7)), 7.88 (m, 1H, arom. H), 8.15 (dd, J = 1.2, 8.0 Hz, 1H-C
(5)), 8.34 (s, 1H-C (2)), 10.64 (s, 1H, CONH). 13C-NMR
(DMSO) 49.18 (NCH2), 121.08, 121.62, 126.35, 127.52,

8
CHANG ET AL.
127.64, 127.68, 129.17, 135.07, 137.72, 148.26, 148.83,
160.72 (C=O), 165.96 (CONH). Anal. Calcd for
C16H12ClN3O2: C 61.25, H 3.86, N 13.39. Found: C 60.91,
H 3.88, N 13.32.
(NPC-TW01) cell line was purchased from the Taiwan Food
Industry Research and Development Institute (Hsinchu, Tai-
wan). All the tumor cell lines were maintained in MEM,
RPMI 1640, or DMEM medium supplied with 10% fetal
bovine serum at 37 C in a humidified atmosphere of 5%
CO2/95% air in the presence of antibiotics.
ꢀ
N-(4-Bromophenyl)-2-(4-oxoquinazolin-3[4H]-yl)acet-
1
ꢀ
amide (8d): Yield 92%. mp 244–245 C. H-NMR (DMSO-
d6) δ: 4.84 (s, 2H, NCH2), 7.48–7.59 (m, 5H, arom. H), 7.72
(d, J = 7.6 Hz, 1H-C (7)), 7.88 (m, 1H, arom. H), 8.12
(dd, J = 1.2, 8.0 Hz, 1H-C (5)), 8.33 (s, 1H-C (2)), 10.64
(s, 1H, CONH). 13C-NMR (DMSO) 49.19 (NCH2), 115.67,
121.47, 121.61, 126.35, 127.52, 127.68, 132.06, 135.07,
138.13, 148.25, 148.82, 160.72 (C=O), 165.98
(C=NOCH3). Anal. Calcd for C16H12BrN3O2: C 53.65, H
3.38, N 11.73. Found: C 53.71, H 3.50, N 11.52.
3.3.2
| Growth inhibition assay (MTT assay)
The colorimetric assay for cellular growth and survival was
performed as described by Hansen et al., with slight modifi-
cations.[20] Pre-determined numbers of cells were seeded in
a 96-well microplate, and designated compounds at various
concentrations were added for the indicated time period. The
MTT-containing solution was added after compound treat-
ment, and conversion of MTT to formazan by metabolically
viable cells was measured by absorbance at 490 nm in a
96-well microtiter plate reader. The percentage of conversion
by mock-treated control cells was used to evaluate the effect
of the chemicals on cell growth and to determine the IC50
values.
N-(4-Methoxyphenyl)-2-(4-oxoquinazolin-3[4H]-yl)acet-
1
ꢀ
amide (8e): Yield: 84%. mp 230–231 C. H-NMR (DMSO-
d6) 4.83 (s, 2H, NCH2), 6.90 (dd, J = 2.0, 6.8 Hz, 2H, arom.
H), 7.48–7.59 (m, 3H, arom. H), 7.73 (dd, J = 0.8, 7.6 Hz,
1H-C (6)), 7.88 (dd, J = 1.6, 7.2 Hz, 1H-C (7)), 8.16 (dd,
J = 1.2, 8.0 Hz, 1H-C (8)), 8.36 (s, 1H-C (2)), 10.33 (s, 1H,
CONH). 13C-NMR (DMSO) 48.75 (NCH2), 55.24 (OCH3)
114.07, 120.68, 121.52, 126.11, 127.27, 127.33, 131.82,
134.65, 148.16, 148.73, 155.46, 160.41 (C=O), 164.99
(C=NOCH3). Anal. Calcd for C17H15N3O3: C 66.01, H
4.89, N 13.58. Found: C 65.90, H 4.91, N 13.55.
3.4 | Cell cycle analysis
Treated or control cells were trypsinized and collected for
overnight fixation with 70% ice-cold ethanol in 1 × phos-
N-(Biphenyl-4-yl)-2-(4-oxoquinazolin-3[4H]-yl)acet-
ꢀ
amide (8f): Yield: 93%. mp 307–308 ꢀC. 1H-NMR (DMSO-
d6) 4.90 (s, 2H, NCH2), 7.30–7.34 (m, 1H, arom. H),
7.42–7.45 (m, 2H, arom. H), 7.56–7.74 (m, 8H, arom. H),
7.85–7.89 (m, 1H-C (7)), 8.17 (dd, J = 1.2, 8.0 Hz, 1H-C
(8)), 8.39 (s, 1H-C (2)), 10.59 (s, 1H, CONH). 13C-NMR
(DMSO) 48.94 (NCH2), 119.56, 121.51, 126.12, 126.35,
127.17, 127.22, 127.31, 127.36, 129.03, 134.70, 135.31,
138.14, 139.61, 148.17, 148.69, 160.42 (C=O), 165.59
(C=NOCH3). Anal. Calcd for C22H17N3O2: C 74.35, H
4.82, N 11. 82. Found: C 74.23, H 4.85, N 11.82.
phate-buffered saline (1 × PBS) at −20 C. After centrifuga-
tion and washing with 1 × PBS for depleting ethanol, cells
were permeabilized with 0.1% triton X-100 in 1 × PBS for
15 min at room temperature and then stained with propidium
iodide (PI) (40 μg/mL) containing RNase A (0.1 mg/mL) in
ꢀ
PBS for 30 min at 37 C. After centrifugation and washing
with 1 × PBS for removing free PI, the samples were
instantly analyzed on
a
Canto II flow cytometer
(BD Biosciences, San Jose, CA) and the distribution of each
cell cycle phase was thus evaluated by the FCS express soft-
ware (De Novo software, Los Angeles, CA).
N-(Naphthalen-2-yl)-2-(4-oxoquinazolin-3[4H]-yl)acet-
1
ꢀ
amide (8g): Yield: 89%. mp 294–295 C. H-NMR (DMSO-
d6) 4.40 (s, 2H, NCH2), 7.39–7.49 (m, 2H, arom. H),
7.56–7.62 (m, 2H, arom. H), 7.73–7.80 (m, 2H, arom. H),
7.85–7.90 (m, 3H, arom. H), 8.17 (dd, J = 1.2, 8.0 Hz, 1H-
C (8)), 8.27 (d, J = 2.0 Hz, 1H, arom. H), 8.41 (s, 1H-C
(2)), 10.70 (s, 1H, CONH). 13C-NMR (DMSO) 48.99
(NCH2), 115.42, 119.77, 121.52, 124.90, 126.12, 126.65,
127.30, 127.35, 127.40, 127.59, 128.70, 129.95, 133.45,
134.68, 136.23, 148.17, 148.7, 160.44 (C=O), 165.83
(C=NOCH3). Anal. Calcd for C20H15N3O2: C 72.94, H
4.59, N 12.76. Found: C 72.62, H 4.60, N 12.71.
4
| CONCLUSIONS
We have synthesized certain oxime, methyloxime, and
amide-containing quinazolinone derivatives for antiprolifera-
tive evaluation against NPC-TW01, Jurkat, and NCI-H226
cancer cells. Among them, N-(naphthalen-2-yl)-2-(4-oxoqui-
nazolin-3[4H]-yl)acetamide (8g) showed the highest activity
against NPC-TW01 cell lines with an IC50 value of
4.78 μM. Flow cytometric analysis indicated that 8g induced
NPC-TW01 cell arrest at the G1/G0 phase in a time- and
concentration- dependent manner. Collectively, this work
provided solid evidence that 8g might be a potential lead
compound capable of inducing nasopharyngeal cancer cell
death and is worthy of advanced development and
investigation.
3.3 | Antiproliferative activity
3.3.1
| Cell lines
Human leukemia (Jurkat) and non-small-cell lung carcinoma
(NCI-H226) were obtained from the American Type Culture
Collection (Rockville, MD). A nasopharyngeal carcinoma

CHANG ET AL.
9
[10] I. L. Chen, K. M. Chang, C. L. Miaw, C. H. Liao, J. J. Chen, T. C. Wang,
Bioorg. Med. Chem. 2007, 15, 6527.
[11] I. L. Chen, J. Y. Chen, P. C. Shieh, J. J. Chen, C. H. Lee, S. H. Juang,
T. C. Wang, Bioorg. Med. Chem. 2008, 16, 7639.
[12] I. L. Chen, J. J. Chen, Y. C. Lin, C. T. Peng, S. H. Juang, T. C. Wang, Eur.
J. Med. Chem. 2013, 59, 227.
ACKNOWLEDGMENT
This work was supported by the Ministry of Science and
Technology of the Republic of China (NSC91-2320-B-
127-005), which is gratefully acknowledged.
[13] L. C. Chen, S. H. Juang, K. M. Chang, C. C. Tzeng, J. J. Chen, I. L. Chen,
T. C. Wang, Chem. Pharm. Bull. 2014, 62, 106.
[14] Y. Y. Xu, S. N. Li, G. J. Yu, Q. H. Hub, H. Q. Li, Bioorg. Med. Chem.
2013, 21, 6084.
[15] I. Khan, A. Ibrar, W. Ahmed, A. Saeed, Eur. J. Med. Chem. 2015, 90, 124.
[16] S. N. Li, Y. Y. Xu, J. Y. Gao, H. R. Yin, S. L. Zhang, H. Q. Li, Bioorg.
Med. Chem. 2015, 23, 3221.
Conflict of interest
All the authors have declared that they have no conflict of
interest.
[17] W. M. Metwally, G. Hegazy, S. S. Eid, A. T. Rabah, E. E. Abou, A. Dalal,
Life Sci. J. 2016, 13, 57.
[18] C. Ito, S. Katsuno, Y. Kondo, H. T. W. Tan, H. Furukawa, Chem. Pharm.
Bull. 2000, 48, 339.
[19] R. M. Silverstein, F. X. Webster, in Spectrometric Identification of Organic
Compounds, 6th ed. (Ed: N. Rose), John Wiley and Sons, New York 1998,
p. 217.
[20] M. B. Hansen, S. E. Nielsen, K. J. Berg, Immunol. Methods 1989,
119, 203.
REFERENCES
[1] M. M. Hsu, S. M. Tu, Cancer 1983, 52, 362.
[2] M. Altun, A. Fandi, O. Dupuis, E. Cvitkovic, Z. Krajina, F. Eschwege, Int.
J. Radiat. Oncol. Biol. Phys. 1995, 32, 859.
[3] G. Niedobitek, Mol. Pathol. 2000, 53, 248.
[4] W. Fang, X. Li, Q. Jiang, Z. Liu, H. Yang, S. Wang, S. Xie, Q. Liu, T. Liu,
J. Huang, W. Xie, Z. Li, Y. Zhao, E. Wang, F. M. Marincola, K. Yao,
J. Transl. Med. 2008, 6, 32.
[5] E. T. Chang, H. O. Adami, Cancer Epidemiol. Biomarkers Prev. 2006, 15,
1765.
[6] C. Hsu, Y. C. Shen, C. C. Cheng, R. L. Hong, C. J. Chang, A. L. Cheng,
Cancer Epidemiol. Biomarkers Prev. 2006, 15, 856.
[7] A. T. Chan, P. M. Teo, T. W. Leung, P. J. Johnson, Cancer 1998, 82,
1003.
[8] C. H. Hsu, C. L. Chen, R. L. Hong, K. L. Chen, J. F. Lin, A. L. Cheng,
Oncology 2002, 62, 305.
How to cite this article: Chang K-M, Chen L-C,
Tzeng C-C, et al. Synthesis and antiproliferative eval-
uation of oxime, methyloxime, and amide-containing
quinazolinones. J Chin Chem Soc. 2018;1–9. https://
doi.org/10.1002/jccs.201700463
[9] J. Ai, W. Li, R. Zeng, Z. Xie, H. Liu, M. Hou, G. Tan, Biomed. Pharmac-
other. 2016, 83, 1022.