Design, synthesis, and evaluation of new 4(3H)-quinazolinone derivatives containing a pyrazole carboxamide moiety

Article abstract of DOI:10.1002/jhet.4334

A total of 15 new 4(3H)-quinazolinone derivatives containing a pyrazole carboxamide moiety were designed and synthesized in this study. The structures of the target compounds were elucidated using ¹H NMR, ¹³C NMR, MS, and elemental a

Full text of DOI:10.1002/jhet.4334

Received: 26 April 2021
DOI: 10.1002/jhet.4334
Revised: 16 June 2021
Accepted: 22 June 2021
A R T I C L E
Design, synthesis, and evaluation of new 4(3H)-
quinazolinone derivatives containing a pyrazole
carboxamide moiety
Xiang Wang1 | Xiaoyu Wang2 | Banghua Zhou1 | Jiefeng Long1 | Pei Li1
1Qiandongnan Engineering and
Abstract
Technology Research Center for
Comprehensive Utilization of National
Medicine, Kaili University, Kaili, China
2School of Chinese Materia Medica,
Shenyang Pharmaceutical University,
Shenyang, China
A total of 15 new 4(3H)-quinazolinone derivatives containing a pyrazole carbox-
amide moiety were designed and synthesized in this study. The structures of the
1
target compounds were elucidated using H NMR, 13C NMR, MS, and elemental
analysis. Then, antifungal activities against Gibberella zeae (G. zeae), Fusarium
oxysporum (F. oxysporum), Cytospora mandshurica (C. mandshurica), Phyto-
phthora infestans (P. infestans), and Pellicularia sasakii (P. sasakii) as well as
antibacterial activities against Ralstonia solanacearum (R. solanacearum) and
Xanthomomu oryzae pv. oryzae (Xoo) were assayed. Bioassay results revealed that
the target compounds have certain antifungal and antibacterial activities. Espe-
cially, compound 5-chloro-1,3-dimethyl-N-(2-(4-oxo-2-(m-tolylamino)quinazolin-3
(4H)-yl)ethyl)-1H-pyrazole-4-carboxamide (7g) exhibited better antibacterial activi-
ties against R. solanacearum (60%) and Xoo (53%) than those of thiodiazole-copper
(52% and 38%, respectively) at 200 μg/ml. This study provided a practical tool for
guiding the design and synthesis of novel and more promising active small mole-
cules of 4(3H)-quinazolinone derivatives containing a pyrazole carboxamide moi-
ety for controlling plant bacterial and fungal diseases.
Correspondence
Pei Li, Qiandongnan Engineering and
Technology Research Center for
Comprehensive Utilization of National
Medicine, Kaili University, Kaili 556011,
China.
Email: pl19890627@126.com
Funding information
In-depth Research Project on High-level
Talented Person of Kaili University,
Grant/Award Number: GCC201803; Kaili
University Doctoral Program, Grant/
Award Number: BS201811; National
Torch Base Project of Qiandongnan Miao-
Dong Medicine Characteristic Industrial,
Grant/Award Number: J[2018]007;
Science and Technology Foundation of
Guizhou Province, Grant/Award Number:
ZK[2021]137; State Key Laboratory
Breeding Base of Green Pesticide and
Agricultural Bioengineering/Key
Laboratory of Ministry of Education,
Grant/Award Number: 2010GDGP0102
1 | INTRODUCTION
costs and the rapidity with which bacterial and fungal
pathogens develop resistance, increased consumer aware-
Plant bacterial and fungal diseases continue to contribute
to heavy global crop losses and restrict the sustainable
development of agriculture despite the best control efforts
of plant pathologists. The preventive and curative use of
synthetic chemical pesticides are the most widely accepted
approaches in plant bacterial and fungal disease manage-
ment [1]. However, apart from the sometimes-prohibitive
ness of the detrimental effects of synthetic chemical pesti-
cides threaten their continued use [2]. Therefore, it is
necessary to develop novel lead molecules with potent bio-
activities against bacterial and fungal diseases.
4(3H)-Quinazolinone and its derivatives, a group of
high-potential and biologically active pharmacophoric
nitrogen-containing heterocyclic molecules, have received
J Heterocyclic Chem. 2021;1–8.
wileyonlinelibrary.com/journal/jhet
© 2021 Wiley Periodicals LLC.
1

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WANG ET AL.
considerable attention in recent years because of their
broad spectrum of pesticide properties, including
antibacterial [3–7], antifungal [3–6], antiviral [8–10], insec-
ticidal [11–13], and herbicidal [14] activities. Our previous
studies have designed and synthesized a series of 4(3H)-
quinazolinone derivatives containing an imines or iso-
xazole moiety (Figure 1), bioassay results showed that the
target compounds exhibited better antibacterial and anti-
fungal activities [3–5]. Meanwhile, pyrazole carboxamide
derivatives are important heterocyclic compounds in the
development of chemical pesticides because of their broad
spectrum of biological activities like antifungal [15–18],
antiviral [19,20], insecticidal [21–23], herbicidal [24,25],
and nematocidal [26] activities. Many recent studies have
been conducted on the synthesis and biological activity of
pyrazole carboxamide derivatives and some pyrazole car-
boxamide derivatives have been developed and commer-
cialized as fungicides in succession, such as penthiopyrad,
furametpyr, penflufen, isopyrazam, and bixafen [15–26].
In view of the facts and to explore the potential lead
molecules with potent antifungal and antibacterial activi-
ties, a series of 4(3H)-quinazolinone derivatives containing
a pyrazole carboxamide moiety are designed and synthe-
sized in the current study. Bioassay results revealed that
the target compounds have certain antifungal and
antibacterial activities. Especially, compound 5-chloro-1,-
3-dimethyl-N-(2-(4-oxo-2-(m-tolylamino)quinazolin-3(4H)-
yl)ethyl)-1H-pyrazole-4-carboxamide (7g) exhibited good
antibacterial activities against Ralstonia solanacearum
(R. solanacearum) and Xanthomomu oryzae pv. oryzae
(Xoo) which were even better than those of thiodiazole-
copper at 200 μg/ml.
using DMSO-d6 as solvent at room temperature. Mass
spectral was conducted on an Agilent 5973 mass spec-
trometer (Agilent Technologies, Santa Clara, CA, USA).
Elemental analysis was performed using an Elemental
Vario-III CHN analyzer (Elementar, Germany). All sol-
vents were dried by standard methods in advance and
distilled before use.
2.2 | General procedure for the
preparation of the key intermediates 6
As shown in Scheme 1, intermediates 6 were synthesized
by a five-step process following previously reported pro-
cedures [4–6].
2.3 | General procedure for the
preparation of the target compounds 7a–7o
To
a
50 ml three-necked round-bottomed flask,
1,3-substituted 5-chloro-1H-pyrazole-4-carboxylic acid
(1.2 mmol), dimethylaminopyridine (DMAP, 1.5 mmol),
and CH2Cl2 (10 ml) were added in order and reacted for
0.5 h at 0ꢀC. After that, intermediates 6 (1.0 mmol), N-
methylmorpholine (20 μl), and carbodiimide hydrochlo-
ride (EDCI, 1.5 mmol) were added to the mixture, and
the reactions were reacted at room temperature for 2.5–
8.0 h. Upon completion of reaction, the redundant
1,3-substituted 5-chloro-1H-pyrazole-4-carboxylic acid
was removed by 10% NaHCO3 solution. Finally, the resi-
dues were filtrated, dried under vacuum, and rec-
rystallized from CH2Cl2/C2H5OH (1:15, v/v) to give pure
products 7a–7o.
2 | MATERIAL AND METHODS
2.1 | General information
2.4 | In vitro antifungal activity test
1H NMR and 13C NMR spectra were recorded on a JEOL-
ECX 500 NMR spectrometer (JEOL Ltd., Japan) operated
IN this study, the in vitro antifungal activities of the target
compounds 7a–7o against Gibberella zeae (G. zeae), Fusar-
ium oxysporum (F. oxysporum), Cytospora mandshurica
(C. mandshurica), Phytophthora infestans (P. infestans),
and Pellicularia sasakii (P. sasakii) were screened at the
concentration of 50 μg/ml by using the mycelium growth
rate method [4–6]. All the target compounds, dissolved in
1 ml DMSO, were mixed with 90 ml potato dextrose agar
(PDA) medium. Then, approximately 4-mm diameter
mycelia dishes, cut from the culture PDA medium, were
inoculated in the middle of the PDA plates. After that, the
inoculated PDA plates were incubated in biochemical
incubator at 28ꢀC for approximately 5 days. DMSO was
served as the negative control, whereas hymexazol and
epoxiconazole widely used for controlling G. zeae,
FIGURE 1 Compounds previously reported with good
antibacterial and antifungal activities

WANG ET AL.
3
SCHEME 1 General synthesis route for the target compounds 7a–7o
F. oxysporum, C. mandshurica, P. infestans, and P. sasakii
[5,27,28] were served as the positive controls. Inhibition
rates I (%) of the target compounds against the test fungus
are calculated by the following formula, where C is the
fungi diameter of the untreated PDA medium, and T is the
fungi diameter of the treated PDA medium. Each experi-
ment was repeated three times.
180 rpm for 24–48 h until the optical density at 595 nm
(OD595) of the bacteria were cultured to 0.6–0.8. DMSO
was served as the negative control, whereas kocide 3000,
thiodiazole copper, and bismerthiazol widely used for
controlling R. solanacearum and Xoo [28–30] were served
as the positive controls. The OD595 values of the cultures
were monitored on a Multiskan Sky 1530 spectrophotom-
eter (Thermo Scientific, Poland). Inhibition rates I (%) of
the target compounds are calculated by the following for-
mula, where C is the corrected turbidity value of the
untreated NB medium, T is the corrected turbidity value
of the treated NB medium.
Inhibition rate I ð%Þ ¼ ðC – TÞ=ðC – 0:4Þꢁ100:
2.5 | In vitro antibacterial activity test
Inhibition rate I ð%Þ ¼ ðC – TÞ=Cꢁ100:
In this study, the in vitro antibacterial activities of the
target compounds 7a–7o against R. solanacearum and
Xoo were screened by the turbidimeter test [4–6].
Approximately 40 μl of solvent nutrient broth
(NB) mediums containing R. solanacearum and Xoo,
respectively, were added to the test tubes containing 5 ml
solvent NB mediums with the testing compounds or the
commercial bactericides concentrations of 200 and
100 μg/ml, respectively. The inoculated test tubes were
incubated in a constant temperature shaker at 28ꢀC and
3 | RESULTS AND DISCUSSION
3.1 | Chemistry
The synthesis of the target compounds 7a–7o is outlined
in Scheme 1. As shown in Scheme 1, the target com-
pounds 7a–7o were synthesized with yields of 64%–85%

4
WANG ET AL.
and determined their structures using 1H NMR, 13C
NMR, MS, and elemental analysis. The physical charac-
ppm) δ: 9.10 (s, 1H, CONH), 8.33 (s, 1H, NH ), 7.97 (d,
1H, J = 7.50 Hz, Ar H), 7.63–7.50 (m, 8H, Ar H), 7.30
(d, J = 8.0 Hz, 1H, Ar H), 7.20–7.10 (m, 3H, Ar H), 4.40
(s, 2H, CH2 ), 3.63 (d, 2H, J = 6.0 Hz, CH2 ), 2.30 (s,
3H, CH3), 2.29 (s, 3H, CH3); 13C NMR (125 MHz,
DMSO-d6, ppm) δ: 163.15, 162.34, 149.68, 149.65, 137.72,
134.86, 129.84, 129.51, 129.47, 129.44, 127.06, 126.90,
125.87, 123.02, 122.98, 122.97, 114.51, 107.46, 40.46, 37.77,
21.03, 13.98; MS (ESI) m/z: 513.3 ([M + H]+); Anal. calcd
for C28H25ClN6O2: C 65.56, H 4.91, N 16.38, found: C
65.21, H 5.02, N 16.25.
5-chloro-1-(4-chlorophenyl)-3-methyl-N-(2-(4-oxo-
2-(p-tolylamino)quinazolin-3(4H)-yl)ethyl)-1H-pyrazole-
4-carboxamide (7e). White solid; yield 83%; 1H NMR
(500 MHz, DMSO-d6, ppm) δ: 8.94 (s, 1H, CONH ),
8.32 (t, J = 6.0 Hz, 1H, NH ), 7.97 (dd, J1 = 1.0 Hz,
J2 = 8.0 Hz, 1H, Ar H), 7.65–7.55 (m, 8H, Ar H), 7.27
(d, J = 6.5 Hz, 1H, Ar H), 7.21–7.15 (m, 2H, Ar H),
4.39 (t, 2H, J = 6.5 Hz, CH2 ), 3.62 (t, 2H, J = 6.0 Hz,
CH2 ), 2.29 (s, 3H, CH3), 2.28 (s, 3H, CH3); 13C
NMR (125 MHz, DMSO-d6, ppm) δ: 163.09, 162.52,
150.00, 148.91, 148.50, 137.37, 136.50, 134.79, 133.92,
132.80, 129.90, 129.30, 127.57, 127.10, 126.96, 125.42,
123.22, 122.80, 117.77, 114.76, 37.87, 37.83, 21.01, 13.96;
MS (ESI) m/z: 547.2 ([M + H]+); Anal. calcd for
C28H24Cl2N6O2: C 61.43, H 4.42, N 15.35, found: C
61.24, H 4.65, N 15.71.
5-chloro-3-methyl-N-(2-(4-oxo-2-(m-tolylamino)quinazolin-
3(4H)-yl)ethyl)-1-phenyl-1H-pyrazole-4-carboxamide(7f).White
solid; yield 79%; 1H NMR (500 MHz, DMSO-d6, ppm) δ: 9.01 (s,
1H, CONH ), 8.34 (t, J = 5.5 Hz, 1H, NH ), 7.99 (d,
J=8.0 Hz, 1H, Ar H), 7.71 (d, J =8.0 Hz, 1H, Ar H), 7.64–7.52
(m, 8H, Ar H), 7.32(d, J=8.0Hz, 1H, Ar H), 7.26–7.20(m, 2H,
Ar H), 6.90 (d, J = 7.5 Hz, 1H, Ar H), 4.38 (t, J = 6.0 Hz, 2H,
CH2 ),3.62(dd,J1=6.5Hz,J2=12.0Hz,2H, CH2 ),2.32(s,
3H, CH3), 2.31 (s, 3H, CH3); 13C NMR (125 MHz, DMSO-d6,
ppm) δ: 163.32, 162.50, 149.84, 148.80, 148.28, 139.99, 137.98,
137.70, 134.81, 129.88, 129.81, 129.47, 128.73, 126.99, 126.96,
125.89, 125.73, 125.52, 124.35, 123.39, 122.93, 120.00, 119.52,
117.87, 114.29, 40.47, 37.95, 21.73, 14.05;MS(ESI)m/z:513.3([M
+ H]+); Anal. calcd for C28H25ClN6O2: C 65.56, H 4.91, N 16.38,
found:C65.86,H5.11,N16.42.
1
teristics, H NMR, 13C NMR, MS, and elemental analysis
data for all synthesized compounds are shown below.
5-chloro-3-methyl-N-(2-(4-oxo-2-anilinoquinazolin-3(4H)-
yl)ethyl)-1-phenyl-1H-pyrazole-4-carboxamide (7a). Off-white
solid; yield 85%; 1H NMR (500 MHz, DMSO-d6,ppm)δ:9.02(s,
1H, CONH ), 8.31 (t, 1H, J = 5.00 Hz, NH ), 7.95 (d, 1H,
J=8.50 Hz, Ar H), 7.78(d, 2H, J =8.00 Hz, Ar H), 7.60–7.47
(m, 6H, Ar H), 7.34–7.26 (m, 3H, Ar H), 7.18 (t, 1H,
J = 7.50 Hz, Ar H), 7.05 (t, 1H, J = 7.00 Hz, Ar H), 4.36 (d,
J=6.50 Hz,2H, CH2 ),3.58(d,J=5.5Hz,2H, CH2 ),2.26
(s, 3H, CH3); 13C NMR (125 MHz, DMSO-d6, ppm)δ: 163.30,
162.51, 149.71, 148.76, 148.31, 140.06, 137.70, 134.83, 129.86,
129.47, 128.89, 126.97, 126.95, 125.90, 125.51, 123.69, 123.44,
122.45, 117.93, 114.44, 40.47, 37.88, 13.98; MS (ESI) m/z: 499.3
([M + H]+),521.2([M + Na]+);Anal.calcdforC27H23ClN6O2:
C64.99,H4.65,N16.84,found:C64.63,H4.87,N17.09.
5-chloro-1,3-dimethyl-N-(2-(4-oxo-2-anilinoquinazolin-
3(4H)-yl)ethyl)-1H-pyrazole-4-carboxamide (7b). White
1
solid; yield 72%; H NMR (500 MHz, DMSO-d6, ppm) δ:
9.16 (s, 1H, CONH ), 8.16 (s, 1H, NH ), 8.00 (d, 1H,
J = 7.50 Hz, Ar H), 7.86 (d, 2H, J = 5.50 Hz, Ar H), 7.75
(s, 1H, Ar H), 7.40–7.33 (m, 3H, Ar H), 7.24 (s, 1H,
Ar H), 7.11 (t, J = 7.0 Hz, 1H, Ar H), 4.37 (t, J = 6.5 Hz,
2H, CH2 ), 3.74 (s, 3H, CH3), 3.60 (d, J = 5.5 Hz, 2H,
CH2 ), 2.25 (s, 3H, CH3); 13C NMR (125 MHz, DMSO-
d6, ppm) δ: 163.59, 162.44, 148.76, 148.22, 148.12, 140.08,
134.83, 128.89, 127.07, 126.96, 125.50, 123.60, 123.39,
122.33, 120.00, 117.87, 40.44, 37.85, 36.47, 13.99; MS (ESI)
m/z: 437.1 ([M + H]+), 459.2 ([M + Na]+); Anal. calcd for
C22H21ClN6O2: C 60.48, H 4.84, N 19.24, found: C 59.99, H
4.93, N 19.35.
5-chloro-1-(4-chlorophenyl)-3-methyl-N-(2-(4-oxo-
2-anilinoquinazolin-3(4H)-yl)ethyl)-1H-pyrazole-4-carboxamide
1
(7c). White solid; yield 82%; H NMR (500 MHz, DMSO-d6,
ppm) δ: 9.07 (s, 1H, CONH ), 8.38 (t, 1H, J = 5.5 Hz,
NH ), 8.02 (d, J = 6.5 Hz, 1H, Ar H), 7.84 (d,
J = 8.0 Hz, 2H, Ar H), 7.68–7.59 (m, 5H, Ar H), 7.39 (t,
J = 8.0 Hz, 2H, Ar H), 7.34 (d, J = 8.0 Hz, 1H, Ar H),
7.26 (t, J = 7.5 Hz, 1H, Ar H), 7.12 (t, J = 7.5 Hz, 1H,
Ar H), 4.44 (t, J = 6.0 Hz, 2H, CH2 ), 3.66 (t,
J = 6.5 Hz, 2H, CH2 ), 2.32 (s, 3H, CH3); 13C NMR
(125 MHz, DMSO-d6, ppm) δ: 163.13, 162.52, 150.00,
148.76, 148.32, 140.05, 136.49, 134.83, 133.93, 129.91,
128.88, 127.58, 127.12, 126.97, 125.51, 123.69, 123.44,
122.45, 117.93, 114.74, 40.47, 37.89, 13.96; MS (ESI) m/z:
533.2 ([M + H]+), 555.2 ([M + Na]+); Anal. calcd for
C27H22Cl2N6O2: C 60.80, H 4.16, N 15.76, found: C 60.55,
H 4.64, N 15.66.
5-chloro-1,3-dimethyl-N-(2-(4-oxo-2-(m-tolylamino)
quinazolin-3(4H)-yl)ethyl)-1H-pyrazole-4-carboxamide
(7g). White solid; yield 75%; 1H NMR (500 MHz,
DMSO-d6, ppm) δ: 9.10 (s, 1H, CONH ), 8.16 (t,
J = 5.0 Hz, 1H, NH ), 8.01 (d, 1H, J = 7.0 Hz, Ar H),
7.75 (d, 1H, J = 8.0 Hz, Ar H), 7.69–7.65 (m, 1H,
Ar H), 7.62 (s, 1H, Ar H), 7.35 (d, 1H, J = 8.0 Hz,
Ar H), 7.27 (dd, 2H, J1 = 8.0 Hz, J2 = 16.5 Hz, Ar H),
6.94 (d, 1H, J = 7.5 Hz, Ar H), 4.37 (t, 2H, J = 7.0 Hz,
CH2 ), 3.76 (s, 3H, CH3), 3.60 (t, 2H, J = 7.0 Hz,
5-chloro-3-methyl-N-(2-(4-oxo-2-(p-tolylamino)quinazolin-
3(4H)-yl)ethyl)-1-phenyl-1H-pyrazole-4-carboxamide (7d).
1
Off-white solid; yield 80%; H NMR (500 MHz, DMSO-d6,
CH2 ), 2.35 (s, 3H, CH3), 2.27 (s, 3H, Ar CH3); 13
C

WANG ET AL.
5
NMR (125 MHz, DMSO-d6, ppm) δ: 163.58, 162.46,
148.78, 148.25, 148.21, 139.98, 137.98, 134.85, 128.74,
127.12, 126.94, 125.52, 124.32, 122.83, 119.42, 112.18,
55.46, 37.90, 36.51, 21.72, 14.04; MS (ESI) m/z: 451.3
([M + H]+), 473.2 ([M + Na]+); Anal. calcd for
C23H23ClN6O2: C 61.26, H 5.14, N 18.64, found: C
60.84, H 5.33, N 18.85.
5-chloro-1-(4-chlorophenyl)-3-methyl-N-(2-(4-oxo-
2-(m-tolylamino)quinazolin-3(4H)-yl)ethyl)-1H-pyrazole-
4-carboxamide (7h). White solid; yield 85%; 1H NMR
(500 MHz, DMSO-d6, ppm) δ: 9.01 (s, 1H, CONH ),
8.36 (t, J = 6.0 Hz, 1H, NH ), 7.94 (dd, J1 = 1.0 Hz,
J2 = 8.0 Hz, 1H, Ar H), 7.69–7.65 (m, 3H, Ar H), 7.61–
7.59 (m, 3H, Ar H), 7.34 (d, 1H, J = 8.0 Hz, Ar H),
7.29–7.24 (m, 2H, Ar H), 6.94 (d, J = 7.5 Hz, 1H,
Ar H), 4.42 (t, J = 7.0 Hz, 2H, CH2 ), 3.65 (t,
J = 6.0 Hz, 2H, CH2 ), 2.34 (s, 3H, CH3); 13C NMR
(125 MHz, DMSO-d6, ppm) δ: 163.13, 162.51, 150.10,
148.79, 148.30, 139.97, 137.97, 136.49, 134.82, 133.94,
129.91, 128.74, 127.57, 127.15, 126.96, 125.52, 124.37,
123.40, 122.95, 119.55, 117.88, 114.62, 37.93, 37.92, 21.72,
14.00; MS (ESI) m/z: 547.2 ([M + H]+); Anal. calcd for
C28H24Cl2N6O2: C 61.43, H 4.42, N 15.35, found: C 61.48,
H 4.72, N 15.35.
118.00, 112.19, 40.36, 37.85, 36.47, 14.02; MS (ESI) m/z:
471.2 ([M + H]+), 493.2 ([M + Na]+); Anal. calcd for
C22H20Cl2N6O2: C 56.06, H 4.28, N 17.83, found: C 56.17,
H 4.37, N 17.94.
5-chloro-N-(2-(4-oxo-2-(p-tolylamino)quinazolin-3
(4H)-yl)ethyl)-1-phenyl-3-(trifluoromethyl)-1H-pyrazole-
4-carboxamide (7k). White solid; yield 72%; 1H NMR
(500 MHz, DMSO-d6, ppm) δ: 8.90 (t, J = 5.5 Hz, 1H,
NH ), 8.87 (s, 1H, CONH ), 8.00 (d, J = 7.0 Hz, 1H,
Ar H), 7.68–7.62 (m, 8H, Ar H), 7.30 (d, J = 8.0 Hz,
1H, Ar H), 7.24–7.20 (m, 2H, Ar H), 4.43 (t, J = 6.0 Hz,
2H, CH2 ), 3.68 (t, J = 6.5 Hz, 2H, CH2 ), 2.34 (s,
3H, CH3); 13C NMR (125 MHz, DMSO-d6, ppm) δ:
162.49, 160.67, 148.86, 148.41, 137.36, 136.87, 134.80,
132.87, 130.77, 130.19, 129.32, 129.02, 126.99, 126.36,
125.45, 123.28, 122.85, 120.00, 117.82, 115.63, 40.63,
37.90, 21.00; MS (ESI) m/z: 567.3 ([M + H]+), 589.3 ([M
+ Na]+); Anal. calcd for C28H22ClF3N6O2: C 59.32, H
3.91, N 14.82, found: C 59.02, H 4.10, N 14.71.
5-chloro-N-(2-(4-oxo-2-(4-trifluoromethoxyanilino)
quinazolin-3(4H)-yl)ethyl)-1-phenyl-3-(trifluoromethyl)-
1H-pyrazole-4-carboxamide (7l). White solid; yield 83%;
1H NMR (500 MHz, DMSO-d6, ppm) δ: 9.08 (s, 1H,
CONH ), 8.90 (t, J = 5.5 Hz, 1H, NH ), 8.01 (d,
J = 8.0 Hz, 1H, Ar H), 7.91 (d, J = 9.5 Hz, 2H, Ar H),
7.64 (d, J = 5.5 Hz, 4H, Ar H), 7.61–7.59 (m, 2H,
Ar H), 7.39 (d, J = 9.0 Hz, 2H, Ar H), 7.35 (d,
J = 8.0 Hz, 1H, Ar H), 7.26 (t, J = 8.0 Hz, 1H, Ar H),
4.44 (t, J = 6.5 Hz, 2H, CH2 ), 3.66 (t, J = 6.5 Hz, 2H,
CH2 ); 13C NMR (125 MHz, DMSO-d6, ppm) δ: 162.42,
160.67, 148.42, 148.07, 144.08, 139.37, 136.83, 134.90,
130.77, 130.19, 128.95, 127.02, 126.33, 125.57, 124.13,
123.78, 123.56, 122.68, 121.73, 118.15, 115.62, 40.54,
37.85; MS (ESI) m/z: 637.3 ([M + H]+), 659.3 ([M
+ Na]+); Anal. calcd for C28H19ClF6N6O3: C 52.80, H
3.01, N 13.19, found: C 52.49, H 2.97, N 13.25.
5-chloro-N-(2-(2-(4-chloroanilino)-4-oxoquinazolin-3
(4H)-yl)ethyl)-1-phenyl-3-(trifluoromethyl)-1H-pyrazole-
4-carboxamide (7m). White solid; yield 75%; 1H NMR
(500 MHz, DMSO-d6, ppm) δ: 8.97 (s, 1H, CONH ),
8.83 (t, J = 5.5 Hz, 1H, NH ), 7.95 (d, J = 7.0 Hz, 1H,
Ar H), 7.78 (d, J = 8.5 Hz, 2H, Ar H), 7.59 (d,
J = 2.0 Hz, 4H, Ar H), 7.55 (dd, J1 = 2.0 Hz,
J2 = 7.0 Hz, 2H, Ar H), 7.37 (d, J = 9.5 Hz, 2H, Ar H),
7.28 (d, J = 9.0 Hz, 1H, Ar H), 7.20 (t, J = 8.0 Hz, 1H,
Ar H), 4.37 (t, J = 6.5 Hz, 2H, CH2 ), 3.61 (t,
J = 6.0 Hz, 2H, CH2 ); 13C NMR (125 MHz, DMSO-d6,
ppm) δ: 162.42, 160.67, 148.47, 148.07, 139.10, 136.84,
134.89, 130.77, 130.20, 128.98, 128.73, 127.33, 127.02,
126.36, 125.56, 123.91, 118.09, 115.59, 40.38, 37.88; MS
(ESI) m/z: 587.3 ([M + H]+), 609.2 ([M + Na]+); Anal.
calcd for C27H19Cl2F3N6O2: C 55.21, H 3.26, N 14.31,
found: C 55.46, H 3.16, N 14.44.
5-chloro-N-(2-(2-(4-chloroanilino)-4-oxoquinazolin-3
(4H)-yl)ethyl)-3-methyl-1-phenyl-1H-pyrazole-
4-carboxamide (7i). White solid; yield 80%; 1H NMR
(500 MHz, DMSO-d6, ppm) δ: 9.17 (s, 1H, CONH ),
8.33 (t, J = 6.0 Hz, 1H, NH ), 7.99 (d, 1H, J = 6.5 Hz,
Ar H), 7.85 (d, J = 6.0 Hz, 2H, Ar H), 7.65 (t,
J = 8.5 Hz, 1H, Ar H), 7.59–7.50 (m, 6H, Ar H), 7.41
(d, J = 8.5 Hz, 2H, Ar H), 7.32 (d, J = 8.5 Hz, 1H,
Ar H), 7.25–7.22 (m, 1H, Ar H), 4.40 (t, J = 6.0 Hz, 2H,
CH2 ), 36.2 (t, J = 5.5 Hz, 2H, CH2 ), 2.29 (s, 3H,
CH3); 13C NMR (125 MHz, DMSO-d6, ppm) δ: 163.30,
162.46, 149.69, 148.52, 148.13, 139.15, 137.69, 134.88,
129.85, 129.48, 128.73, 127.22, 126.99, 125.55, 119.99,
118.05, 114.37, 40.46, 37.88, 14.00; MS (ESI) m/z: 533.2
([M + H]+); Anal. calcd for C27H22Cl2N6O2: C 60.80, H
4.16, N 15.76, found: C 60.74, H 4.22, N 15.75.
5-chloro-N-(2-(2-(4-chloroanilino)-4-oxoquinazolin-3
(4H)-yl)ethyl)-1,3-dimethyl-1H-pyrazole-4-carboxamide
1
(7j). White solid; yield 83%; H NMR (500 MHz, DMSO-
d6, ppm) δ: 9.26 (s, 1H, CONH ), 8.13 (t, J = 5.5 Hz,
1H, NH ), 8.00 (dd, J1 = 1.0 Hz, J2 = 8.0 Hz, 1H,
Ar H), 7.89 (s, 1H, Ar H), 7.87 (d, J = 2.5 Hz,
1H, Ar H), 7.68–7.65 (m, 1H, Ar H), 7.42 (d,
J = 8.5 Hz, 2H, Ar H), 7.34 (d, J = 7.5 Hz, 1H, Ar H),
7.25 (t, J = 7.5 Hz, 1H, Ar H), 4.35 (t, 2H, J = 6.5 Hz,
CH2 ), 3.73 (s, 3H, CH3), 3.58 (t, 2H, J = 6.5 Hz,
CH2 ), 2.22 (s, 3H, CH3); 13C NMR (125 MHz,
DMSO-d6, ppm) δ: 163.60, 162.40, 148.50, 148.11, 148.06,
139.14, 134.88, 128.72, 127.16, 127.12, 126.96, 123.61,

6
WANG ET AL.
3-chloro-N-(2-(4-oxo-2-(4-trifluoromethoxyanilino)
134.95, 131.62, 129.37, 128.63, 128.12, 127.21, 126.93,
125.57, 123.77, 123.27, 117.97, 115.79, 107.33, 107.23,
40.91, 37.61; MS (ESI) m/z: 556.2 ([M + H]+); Anal. calcd
for C25H18Cl3N7O2: C 57.70, H 3.68, N 18.84, found: C
57.54, H 3.52, N 18.96.
quinazolin-3(4H)-yl)ethyl)-1-(pyridin-2-yl)-1H-pyrazole-
5-carboxamide (7n). White solid; yield 64%; 1H NMR
(500 MHz, DMSO-d6, ppm) δ: 9.33 (s, 1H, CONH ),
8.92 (s, 1H, NH ), 8.52 (d, J = 3.5 Hz, 1H, Ar H), 8.23
(d, J = 7.0 Hz, 1H, Ar H), 8.00 (d, J = 2.0 Hz, 1H,
Ar H), 7.69–7.64 (m, 4H, Ar H), 7.36–7.25 (m, 4H,
Ar H), 7.04 (s, 1H, Ar H), 4.30 (d, J = 7.0 Hz, 2H,
CH2 ), 3.50 (d, J = 5.5 Hz, 2H, CH2 ); 13C NMR
(125 MHz, DMSO-d6, ppm) δ: 162.36, 158.60, 149.09,
148.38, 147.84, 147.65, 143.86, 139.99, 139.75, 139.24,
134.95, 128.65, 127.18, 126.93, 125.61, 123.86, 122.99,
121.65, 119.74, 118.05, 107.35, 40.53, 37.60; MS (ESI) m/z:
604.3 ([M + H]+), 626.2 ([M + Na]+); Anal. calcd for
C26H18Cl2F3N7O3: C 54.79, H 3.36, N 17.02, found: C
54.38, H 3.38, N 16.99.
3.2 | Biological evaluations
The preliminary in vitro antifungal and antibacterial
activities of the target compounds 7a–7o were deter-
mined and the results are listed in Tables 1 and 2. As
suggested in Table 1, most of the target compounds
exhibited antifungal activities against G. zeae,
F. oxysporum, C. mandshurica, P. infestans, and P. sasakii
to some extent at 50 μg/ml, unfortunately, lower than
those of hymexazol and epoxiconazole. Meanwhile,
Table 2 showed that most of the target compounds
exhibited certain antibacterial activities against
R. solanacearum and Xoo at 200 and 100 μg/ml. It was
interesting that compound 5-chloro-1,3-dimethyl-N-
(2-(4-oxo-2-(m-tolylamino)quinazolin-3(4H)-yl)ethyl)-1H-
pyrazole-4-carboxamide (7g) showed better antibacterial
activities against R. solanacearum (60%) and Xoo (53%) at
200 μg/ml than those of thiodiazole-copper, to the con-
trary, lower than those of kocide 3000 and bismerthiazol.
3-chloro-N-(2-(2-(4-chloroanilino)-4-oxoquinazolin-3
(4H)-yl)ethyl)-1-(pyridin-2-yl)-1H-pyrazole-
5-carboxamide (7o). White solid; yield 83%; 1H NMR
(500 MHz, DMSO-d6, ppm) δ: 9.33 (s, 1H, CONH ),
8.85 (s, 1H, NH ), 8.51 (s, 1H, Ar H), 8.22 (s, 1H,
Ar H), 7.96 (d, J = 13.0 Hz, 1H, Ar H), 7.66 (s, 2H,
Ar H), 7.55 (d, J = 8.5 Hz, 1H, Ar H), 7.32 (d,
J = 9.5 Hz, 3H, Ar H), 7.24 (d, J = 5.5 Hz, 1H, Ar H),
7.03 (s, 1H, Ar H); 13C NMR (125 MHz, DMSO-d6, ppm)
δ: 158.62, 149.09, 148.39, 147.65, 139.97, 139.76, 134.95,
TABLE 1 Antifungal activities of the target compounds 7a-7o against Gibberella zeae, Fusarium oxysporum, Cytospora mandshurica,
Phytophthora infestans, and Pellicularia sasakii at 50 μg/ml
Inhibition rate (%)
Compounds
G. zeae
F. oxysporum
5.33 0.90
6.33 1.00
8.00 1.18
6.67 1.34
7.33 0.91
6.67 1.03
4.33 0.99
8.00 1.18
5.33 1.13
11.33 1.46
0
C. mandshurica
1.20 0.84
0
P. infestans
16.51 0.59
26.91 1.29
11.93 0.62
10.70 1.51
9.79 0.50
8.26 0.40
4.89 0.49
6.73 0.67
25.08 2.00
26.91 1.18
8.26 0.76
3.98 1.05
4.59 0.87
16.51 1.01
21.41 0.89
51.21 5.96
96.07 4.76
P. sasakii
7a
4.45 0.77
7.12 1.39
2.08 0.67
9.50 1.09
6.53 0.65
18.10 1.55
13.95 0.93
8.31 0.80
7.12 0.60
8.90 0.99
0
0
7b
0
7c
0.00 0.52
8.98 0.54
1.80 0.52
0.60 0.52
0.30 0.53
11.08 0.56
2.10 0.54
0.90 0.77
0.60 0.52
5.69 0.55
1.20 0.72
3.59 0.38
0.90 0.33
49.61 7.84
100.00 3.96
0
7d
0
7e
0
7f
0
7g
0
7h
0
7i
3.56 0.87
12.62 1.97
0
7j
7k
7l
1.19 0.64
1.78 0.59
4.45 0.63
2.67 0.63
55.54 3.90
100.00 4.90
4.67 0.89
3.00 1.02
3.67 0.85
3.33 1.01
56.12 4.10
100.00 2.89
0
7m
0
7n
10.03 0.56
19.09 0.74
68.22 2.41
87.62 3.06
7o
Hymexazol
Epoxiconazole

WANG ET AL.
7
TABLE 2 Antibacterial activities of
the target compounds 7a–7o against
Ralstonia solanacearum and
Inhibition rate (%)
R. solanacearum
Xoo
Xanthomomu oryzae pv. oryzae at 200
and 100 μg/ml
Compounds
200 μg/ml
25 4.50
29 3.05
100 μg/ml
200 μg/ml
14 2.11
38 1.11
0
100 μg/ml
7a
15 4.12
18 3.08
10 2.1
7b
8
1.32
7c
9
2.01
7
1.49
0
7d
25 4.12
25 3.16
51 1.55
60 2.34
47 3.14
10 2.02
18 2.30
39 3.11
40 2.50
11 3.15
36 1.56
28 3.12
55 2.00
53 1.48
0
12 2.18
7e
25 2.01
7f
35 1.59
7g
32 2.07
7h
0
7i
7
0.83
5
1.14
0
0
7j
32 2.13
24 2.37
45 2.85
20 2.41
25 2.15
43 1.50
100 1.25
52 1.18
/
26 3.12
10 3.16
20 2.39
18 2.04
35 1.17
48 1.95
64 3.00
11 3.58
25 3.55
0
0
7k
20 1.57
33 1.99
34 2.24
0
7l
7m
4
5
2.95
2.72
7n
7o
23 2.09
68 3.13
31 3.21
/
0
Kocide 3000
Thiodiazole copper
Bismerthiazol
0
38 3.45
73 0.68
32 2.51
52 1.61
4 | CONCLUSION
Laboratory of Ministry of Education, grant number
2010GDGP0102; Kaili University Doctoral Program,
grant number BS201811.
In conclusion, a total of 15 new 4(3H)-quinazolinone
derivatives containing a pyrazole carboxamide moiety
were synthesized and investigated for their in vitro anti-
fungal and antibacterial activities. The preliminary bio-
assay results showed that the target compounds
exhibited certain inhibitory activity against the test bac-
teria and fungus. Especially, compound 7g exhibited
good antibacterial activities against R. solanacearum
and Xoo, which were even better than those of
thiodiazole-copper at 200 μg/ml. To the best of our
knowledge, this is the first report on the antifungal and
antibacterial activities of this series of new 4(3H)-
quinazolinone derivatives containing a pyrazole carbox-
amide moiety.
DATA AVAILABILITY STATEMENT
The data that support the findings of this study are avail-
able from the corresponding author upon reasonable
request.
ORCID
Pei Li https://orcid.org/0000-0001-7276-6337
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How to cite this article: X. Wang, X. Wang,
B. Zhou, J. Long, P. Li, J Heterocyclic Chem 2021, 1.
https://doi.org/10.1002/jhet.4334