Synthesis and Biological Activity of Amide Compounds Containing Pyrazole Mandelic Acid Groups

Article abstract of DOI:10.1002/jhet.2862

A series of novel mandelic acid pyrazole amide compounds were synthesized and characterized. Moreover, their antiviral activities against tobacco mosaic virus were evaluated. The bioassay results indicated that as-prepared compounds showed antiviral activity against tobacco mosaic virus at a concentration of 500?g/mL. The curative activity of compound 4g is 59.5%, which was comparable with positive control (Ningnanmycin, 61.0%). The inactivation activity of the compound 4l exhibited 80.1%, which was higher than that of Ningnanmycin (75.0%), and its protective activity was 88.7%, which is comparable with that of Ningnanmycin (90.7%).

Full text of DOI:10.1002/jhet.2862

Month 2017
Synthesis and Biological Activity of Amide Compounds Containing
Pyrazole Mandelic Acid Groups
*
Yan Xie, Xiang-Hui Ruan, Hua-Yu Gong, Yi-Hui Wang, Xiao-Bin Wang, Ju-Ping Zhang, Qin Li, and Wei Xue
State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green
Pesticide and Agricultural Bioengineering, Ministry of Education, Center for Research and Development of Fine
Chemicals, Guizhou University, Guiyang 550025, People’s Republic of China
*
E-mail: wxue@gzu.edu.cn
Additional Supporting Information may be found in the online version of this article.
Received October 31, 2016
DOI 10.1002/jhet.2862
Published online 00 Month 2017 in Wiley Online Library (wileyonlinelibrary.com).
A series of novel mandelic acid pyrazole amide compounds were synthesized and characterized.
Moreover, their antiviral activities against tobacco mosaic virus were evaluated. The bioassay results indi-
cated that as-prepared compounds showed antiviral activity against tobacco mosaic virus at a concentration
of 500 g/mL. The curative activity of compound 4g is 59.5%, which was comparable with positive control
(Ningnanmycin, 61.0%). The inactivation activity of the compound 4l exhibited 80.1%, which was higher
than that of Ningnanmycin (75.0%), and its protective activity was 88.7%, which is comparable with that
of Ningnanmycin (90.7%).
J. Heterocyclic Chem., 00, 00 (2017).
INTRODUCTION
diseases of leaf spot, rust, powdery mildew, downy
mildew, epidemic of wheat, rice, tobacco, and so
on [17].
The management of plant diseases is highly important
for preventing the losses in agricultural economy [1].
In current agricultural system, various commercial
agents have been applied in effective control of the
plant diseases. However, many disadvantages have
emerged with wide using of pesticide, such as the
increase of the resistance and residues [2], which are
harmful to humans and animals [3,4]. A growing
number of amide compounds reveal certain
bacteriostatic [5], insecticidal [6], acaricidal [7], anti-
plant viral [8], anti-inflammatory [9], and anticancer
activities [10], emerged in pharmaceutical chemistry.
Mandelic acid, as
a significant intermediate of
Methenamine mandelate, Cyclanelate and Cemandil
sodium salt is widely used in the process of medicinal
creation. Mandipropamid reported by Syngenta is the first
commercial pesticide containing mandilic acid moiety.
Their special endurance washed by water and long
lasting-effects has attracted
attentions to explore mandelic acid derivatives.
a growing number of
Based on the aforementioned founds, we aim to
introduce the pyrazole amide containing mandilic acid,
and a series of compounds were synthesized. Using the
half-leaf method, their anti-tobacco mosaic virus (TMV)
activities were evaluated, and the results of bioassay
showed that most of the compounds exhibited good anti-
TMV activities at 500 μg/mL.
For example, as
a
kind of sorafenib analogs,
a new series of benzothiazole amide derivatives, which
were synthesized by Damasy, can be used as potent
anticancer agents [11]. As Deng reports N-(3-
methoxyphenyl)-3-(tert-butyl)-1-phenyl-1H-pyrazole-5-
carboxamide derivatives exhibit efficient insecticidal
activities [12]. In addition, pyrazole derivatives, as a
family type of heterocyclic compounds, which attracts
considerable attention because of their good biological
activities including bacteriostatic [13], anticancer [14],
anti-inflamatory [15], and antitumor activities [16]. For
instance, commercial agent thiazole amine ester which
was developed by basf (Germany) in 1993 is a kind
of fungicides containing pyrazole. It can be used to treat
RESULTS AND DISCUSSION
Chemistry.
The detailed synthetic route for the
preparation of amide compounds bearing pyrazole
mandelic acid groups are summarized in Scheme 1.
The
intermediate
5-amino-1-phenyl-1H-pyrazole-4-
carbonitrile(2) were synthesized from substituted
phenylhydrazine and 2-ethoxy methylene malononitrile
© 2017 Wiley Periodicals, Inc.

Y. Xie, X.-H. Ruan, H.-Y. Gong, Y.-H. Wang, X.-B. Wang, J.-P. Zhang, Q. Li, and W. Xue
Vol 000
Scheme 1. Synthetic route to compounds 4a–4p.
(1) reacting with methanol and triethylamine as a solvent
under refluxing condition; the 5-amino-1-substituted
phenylhydrazine-1H-pyrazole-4-methyl formate (3) were
synthesized from cyclization with hydrochloric acid
methanol solution under reflux condition. Subsequently,
1-subsituted phenyl-5-(2–subsituted phenyl-2-hydroxy
ethyl amide)-N-methyl-1H-pyrazole-4-acetyl methylamine
(4) were synthesized from 5-amino-1-subsituted
phenyl-1H-pyrazole-4-methyl formate (3) and 2-formyl
chloride-2-oxalyl chloride methyl benzene by heating
reflux about 1 h, then add methylamine water solution
under cooling to mix about 14 h. The structure of all
NCH3 group. The chemical shifts at 172.4, 74.1, and
20.0 ppm in 13C NMR confirmed the existence of
C═O, CH2, and NCH3 groups; furthermore, all the
target compounds were also confirmed by MS, which
were in accordance with their molecular weight.
Antiviral activity. For us to make an evaluation of the
antiviral properties of 1-subsituted -phenyl-5-(2–
substituted-phenyl-2-hydroxy ethyl amide)-N-methyl-1H-
pyrazole- 4-acetyl methylamine (4a–4p), the bioassays
against TMV have been carried out according to the half-
leaf method with the commercial agent Ningnanmycin as
a
contrast [17]. The antiviral activities (curative,
1
compounds 4a–4p was confirmed by IR, H NMR, 13C
inactivation, and protection) of all the compounds are
listed in Table 1.
NMR, MS, and elemental analysis. The IR spectrum of
4a displayed absorption bands between 3365 and
3304 cmꢀ1 corresponding to N─H, 1714–1624 cmꢀ1
corresponding to C═O of mandelic acid and
3267 cmꢀ1 corresponding to C═O of methanamide.
As shown in Table 1, the anti-TMV bioassay in vitro
indicated that most of the tested compounds displayed
good antiviral activity. Particularly compounds 4g
curative activity was 59.5%, slightly lower than the
contrast agent Ningnanmycin (61.0%). Compound 4l
passivation activity was 80.1%, higher than that of
Ningnanmycin (75.0%). However, its protection
activity was 88.7%, which was closed to the contrast
1
The H–NMR spectrum of 4a in DMSO-d6 displayed a
singlet at δ 10.17 ppm that corresponds to H─N─C═O
functionality, a singlet at δ 7.49 ppm to pyrazole ring
ꢀNH proton, and a singlet at δ 6.59 ppm that
corresponds to OH functionality, a singlet that appeared
at δ 3.61 ppm was observed because of the protons of
agent
building
Ningnanmycin
(90.0%)
at
a
concentration of 500 g/mL. we found that the
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2017
Synthesis and Biological Activity of Amide Compounds Containing Pyrazole
Mandelic Acid Groups
Table 1
EXPERIMENTAL
Curative, protective and inactivation effect of compounds against TMV
in vivo.
General procedures. Melting points were recorded on
an XT-4 binocular microscope melting point apparatus
(Beijing Tech Instruments Co., Beijing, China) and were
reported uncorrected. By using thin-layer chromatography
on silica gel GF254 to monitor reaction. Infrared spectra
were obtained in KBr on a Bruker VECTOR 22 spectrom-
eter. Mass spectral were recorded on an Agilent 5973
organic mass spectrometer. 1H and 13C NMR (solvent
DMSO-d6) spectra were determined on a JEOL-ECX 500
NMR spectrometer at room temperature using TMS as an
internal standard. Unless otherwise noted, all solvents did
not need to dry before use. All common reagents and sol-
vents were obtained from commercial supplies without
more purification.
Compounds Curative (%) Inactivation (%) Protective (%)
4a
4b
4c
4d
4e
4f
4g
4h
4i
4j
4k
4l
4m
4n
4o
4p
41.4
47.7
31.3
20.0
18.0
12.5
59.5
29.8
24.5
43.3
20.0
33.3
5.0
65.2
70.7
48.5
24.0
25.0
52.3
60.0
68.9
25.9
66.3
45.6
80.1
44.8
36.2
42.6
46.9
75.0
75.5
80.0
56.1
70.0
30.0
77.5
70.0
71.1
63.2
71.3
79.5
88.7
74.6
58.3
69.7
56.0
90.0
30.0
37.2
33.0
61.0
General procedure for synthesis of 1-substituted phenyl-
5-(2-substituted phenyl-2-hydroxy ethyl amide)-N-methyl-
Ningnanmycin
1H–pyrazole-4–acetyl methylamin 4a–4p.
A solution of
aAverage of three replicates.
bThe commercial agricultural ningnanmycin was used for comparison of
antiviral activity.
5-amino-1-substituted
phenyl-1H-pyrazole-4-methyl
formate 3(0.5 g) was added to the reaction bottle, rapidly
heat to 140°C for 1 h, thin-layer chromatography tracking
(PE/EA = 3/1), then 2 mL methylamine solution was
added in cooling temperature, stirring for 14 h, finally
adding water (2 mL) and methylene chloride (5 mL × 2)
to dilute. Then using 1 mol/L HCl to adjust pH of
solution to 2, filtering, elution with PE/EA = 2:1 lotion,
get products by decompression drying (yield of about
50%). using similarity methods to synthesize 4b–4p.
1-(4-fluorophenyl)-5-(2-hydroxy-2-phenylacetamido)-N-methyl-
1H-pyrazole-4-carboxamidex (4a). A white powder; yield
48.0%; M.p. 218–219°C; IR (KBr) ν: 3365, 3304, 3267,
1714, 1624, 1593, 1514, 1487, 1458, 1398, 1377, 1276,
1224, 1186, 1157, 1089, 1060, 968, 848, 837, 719. H
NMR (500 MHz, DMSO-d6) δ: 8.0(s, 1H, NH), 7.89(d,
1H, CH), 7.33–7.27(m, 8H, Ph-H), 7.03(m, 2H, OH,
NH), 4.95(s, 1H, CH), 2.68(s, 3H, NCH3); 13C NMR
(125 MHz, DMSO-d6) δ: 172.4, 162.5, 162.4, 160.3,
139.6, 135.6, 128.5, 128.1, 127.3, 125.9, 125.8, 116.1,
115.9, 74.1, 20.0. Anal. Calcd. for C19H17FN4O3: C
61.95, H 4.65, N 15.21; Found C 61.47, H 4.10, N
15.01.
structure–activity relationships of the aimed compounds
were pretty obvious. When R1 was substituted by
p-chlorinated or p-fluorine, the antiviral activities
against TMV of corresponding compounds were found
to exhibit better than that of others. For example, the
inhibition rates against TMV of compounds 4b
(R1 = 4-F) and 4j (R1 = 4-Cl) were higher than that
of the compound 4i (R1 = 4-Br). Meantime, when R2
were substituted by 2-Cl or H, the antiviral activities
against TMV of corresponding compounds were found
to exhibit better than that of others. For example, the
inhibition rates against TMV of compounds 4b
(R2 = 2-Cl) and 4a (R2 = H) were higher than that of
the compound 4c (R2 = 4-Cl) and 4d (R2 = 2-F).
1
CONCLUSIONS
In summary, we have synthesized 16 novel 1-substituted
phenyl-5-(2-substituted phenyl-2-hydroxy ethyl amide)-N-
methyl-1H–pyrazole-4–acetyl methylamin derivatives. All
compounds were evaluated for their antiviral activity
against TMV in vivo. The preliminary biological tests
showed that parts of compounds exhibited better antiviral
activity against TMV. Among these compounds, the
curative activity of 4g was 59.5%, slightly lower than the
commercial agent Ningnanmycin (61.0%). In addition,
the passivation activity of 4l was 80.1%, which was
better than that of Ningnanmycin (75.0%), and its
protection activity was 88.7%, which was closed to the
contrast Ningnanmycin (90.0%).
1-(4-fluorophenyl)-5-(2-(2-chlorophenyl)-2-hydroxyacetamido)-
N-methyl-1H-pyrazole-4-carboxamide (4b).
powder; yield 50.1%; M.p. 227–228°C; IR (KBr) ν:
3365, 3263, 1716, 1624, 1591, 1514, 1489, 1456, 1396,
1377, 1222, 1155, 1082, 1035, 968, 840, 746.1H NMR
(500 MHz, DMSO-d6) δ: 10.26(s, 1H, NH), 8.05(s, 1H,
A
white
Py-H), 7.52(d,
J = 8.6 Hz, 1H, Ph-H), 7.38(d,
J = 3.5 Hz, 4H, Ph-H), 7.45(d, 2H, Ph-H), 7.35(m, 2H,
Ph-H ), 6.80(s, 1H, OH), 5.07(s, 1H, CH), 2.72(s, 3H,
OCH3); 13C NMR (125 MHz, DMSO-d6) δ: 171.7,
162.4, 139.5, 138.5,135.6, 133.1, 130.1, 129.8, 129.7,
127.6, 126.5, 126.4, 116.4, 116.2, 112.6, 70.9, 26.1.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Y. Xie, X.-H. Ruan, H.-Y. Gong, Y.-H. Wang, X.-B. Wang, J.-P. Zhang, Q. Li, and W. Xue
Vol 000
Anal. Calcd. for C19H16ClFN4O3: C 56.65, H 4.00, N
13.91; Found C 56.47, H 3.87, N 13.97.
Ph-H), 6.76(s, 1H, OH), 5.01(s, 1H, CH), 2.7 0(s, 3H,
NCH3); 13C NMR (125 MHz, DMSO-d6) δ: 172.1,
162.3, 140.4, 139.8, 139.5, 137.1, 132.9, 131.2, 131.0,
129.1, 128.6, 126.5, 122.7, 121.9, 112.4, 73.3, 26.0.
Anal. Calcd. for C19H16BrClN4O3: C 49.21, H 3.48, N
12.08; Found C 48.85, H 3.37, N 12.42.
1-(4-fluorophenyl)-5-(2-(4-chlorophenyl)-2-hydroxyacetamido)-
N-methyl-1H-pyrazole-4-carboxamide (4c). A white powder;
yield 49.3%; M.p. 217–219°C; IR (KBr) ν:3363, 1707,
1616, 1577, 1489, 1458, 1398, 1278, 1224, 1070, 1014.1H
1
NMR (500 MHz, DMSO-d6) H NMR (500 MHz, DMSO-
1-(3-bromophenyl)-5-(2-hydroxy-2-phenylacetamido)-N-methyl-
D6)δ: 10.21(s, 1H, NH), 8.01(s, 1H, Py-H), 7.92(d,
J = 4.6 Hz, 1H, NH), 7.32–7.28(m, 5H, Ph-H), 7.04(t,
J = 8.9 Hz, 2H, Ph-H), 6.68(d, J = 4.6 Hz, 1H, OH), 5.00(t,
1H, CH), 2.70(s, 3H, OCH3); 13C NMR (125 MHz,
DMSO-d6) δ: 172.1, 162.4, 160.5, 139.5, 136.7, 135.3,
132.9, 129.0, 128.5, 126.1, 116.1, 116.0, 112.6, 73.3, 26.0.
Anal. Calcd. for C19H16ClFN4O3: C 56.65, H 4.00, N
13.91; Found C 56.34, H 3.70, N 14.14.
1H-pyrazole-4-carboxamide (4g). A gray powder, yield
51.6%; M.p. 203–205°C; IR (KBr) ν: 3323, 3280,
1701, 1622, 1593, 1575, 1558, 1479, 1456, 1408,
1315, 1274, 1186, 1151, 1093, 1058, 750, 717, 696.
1H NMR (500 MHz, DMSO-d6) δ: 10.39(s, 1H, NH),
8.12(s, 1H, Py-H), 7.66(s, 1H, NH), 7.52(d,
J = 8.6 Hz, 1H, Ph-H), 7.38(d, J = 3.5 Hz, 2H, Ph-
H), 7.33–7.25(m, 6H, Ph-H), 6.72(s, 1H, OH), 5.01(s,
1H, CH), 2.69(s, 3H, NCH3); 13C NMR (125 MHz,
DMSO-d6) δ: 173.1, 172.5, 162.3, 140.6, 140.1, 131.2,
130.9, 128.6, 128.4, 127.4, 127.0, 126.4, 122.5, 121.9,
112.6, 74.0, 26.1. Anal. Calcd. for C19H17BrN4O3: C,
53.16; H, 3.99; N, 13.05; Found C 52.75, H 3.90, N
12.87.
1-(4-fluorophenyl)-5-(2-(2-fluorophenyl)-2-hydroxyacetamido)-
N-methyl-1H-pyrazole-4-carboxamide (4d).
A
yellow
powder; yield 52.4%; M.p. 222–224°C; IR (KBr) ν:
3365, 3304, 3265, 1716, 1624, 1593, 1516, 1489, 1458,
1
1398, 1224, 840, 758. H NMR (500 MHz, DMSO-d6)
δ: 10.27(s, 1H, NH), 8.03(dd, J = 9.8,0.8 Hz, 1H, Py-H),
7.92(d, J = 4.6 Hz, 1H, NH), 7.41(dd, J = 8.1, 4.9 Hz,
2H, Ph-H), 7.31(t, J = 6.3 Hz, 2H, Ph-H), 7.20(t,
J = 8.6 Hz, 2H, Ph-H), 7.14(t, J = 6.3 Hz, 2H, Ph-H),
6.75(s, 1H, OH), 5.21(d, J = 5.2 Hz, 1H, CH), 2.71(d,
J = 4.6 Hz, 3H, NCH3); 13C NMR (125 MHz,
DMSO-d6).δ: 171.8, 162.4, 139.5, 136.8, 135.6, 130.5,
129.7, 126.3, 126.2, 124.7, 120.0, 116.3, 115.8, 112.5,
68.0, 26.1. Anal. Calcd. for C19H16F2N4O3: C 59.07, H
4.17, N 14.50; Found C 58.78, H 3.92, N 14.22.
1-(4-bromophenyl)-5-(2-hydroxy-2-phenylacetamido)-N-methyl-
1H-pyrazole-4-carboxamide (4h). A white powder, yield
51.3%; M.p. 183–185°C; IR (KBr) ν: 3444, 3347, 3327,
1716, 1670, 1624, 1600, 1577, 1558, 1483, 1456, 1394,
1273, 1060, 966, 827, 715. 1H NMR (500 MHz,
DMSO-d6) δ: 10.37(s, 1H, NH), 8.05(s, 1H, Py-H),
8.13–7.97 (m, 2H), 7.45–7.14 (m, 9H), 4.98(s, 1H, CH),
2.47(s, 3H, NCH3); 13C NMR (125 MHz, DMSO-d6) δ:
172.4, 162.4, 140.3, 138.3, 136.6, 132.2, 128.5, 128.2,
127.3, 125.6, 120.9, 112.7, 74.0, 26.1, 24.8, 21.6. Anal.
Calcd. for C19H17BrN4O3: C 53.16, H 3.99, N 13.05;
Found C 53.44, H 4.03, N 12.97.
1-(3-bromophenyl)-5-(2-(2-fluorophenyl)-2-hydroxyacetamido)-
N-methyl-1H-pyrazole-4-carboxamide (4e).
A
white
powder; yield 51.2% ; M.p. 217–219°C; IR (KBr) ν:
3232, 3296, 1714, 1616, 1593, 1506, 1485, 1456, 1398,
1375, 1273, 1222, 1147, 1101, 1068, 970, 869, 752. H
1-(4-bromophenyl)-5-(2-(2-chlorophenyl)-2-hydroxyacetamido)-
1
N-methyl-1H-pyrazole-4-carboxamide (4i).
A
white
NMR (500 MHz, DMSO-d6) δ: 10.39(s, 1H, NH),
8.09(s, 1H, Py-H), 8.01(q, J = 4.3 Hz, 1H), 7.66(t,
J = 4.6 Hz, 1H, NH), 7.55(d, J = 8.0 Hz, 1H, Ph-H),
7.43(d, J = 9.2 Hz, 1H, Ph-H), 7.37–7.26(m, 3H, Ph-H),
7.16(m, J = 8.0 Hz, 2H, Ph-H), 6.82(d, J = 5.2 Hz, 1H,
OH), 5.23(d, 1H, CH), 2.72(s, 3H, NCH3); 13C NMR
(125 MHz, DMSO-d6) δ: 166.2, 162.3, 159.6, 140.5,
139.9, 131.4, 131.1, 129.7, 126.5, 124.9(d), 122.8,
121.9, 120.0, 115.9, 115.7, 112.4, 68.0, 26.1. Anal.
Calcd. for C19H16BrFN4O3: C 51.02, H 3.61, N 12.53;
Found C 50.84, H 3.45, N 12.71.
powder; yield 52.7%; M.p. 203–205°C; IR (KBr) ν:
3342, 3165, 1697, 1625, 1593, 1573, 1492, 1471, 1458,
1408, 1390, 1296, 1265, 1188, 1083, 1070, 1031, 1006,
958, 827, 754. 1H NMR (500 MHz, DMSO-d6) δ:
10.32(s, 1H, NH), 8.06(s, 1H, Py-H), 8.00(d, J = 4.3 Hz,
1H), 7.55(d, J = 8.6 Hz, 2H, Ph-H, NH), 7.38–7.36(m,
4H, Ph-H), 7.28(t, J = 5.2 Hz, 2H, Ph-H), 6.82(s, 1H,
OH), 5.35(s, 1H, CH), 2.72(s, 3H, NCH3); 13C NMR
(125 MHz, DMSO-d6) δ: 171.6, 162.4, 139.8, 138.5,
133.2, 132.6, 132.4, 130.3, 130.0, 129.7, 127.6, 126.4,
126.0, 121.1, 112.6, 70.9, 26.1. Anal. Calcd. for
C19H16BrClN4O3: C 49.21, H 3.48, N 12.08; Found C
49.30, H 3.23, N 11.84.
1-(3-bromophenyl)-5-(2-(4-chlorophenyl)-2-hydroxyacetamido)-
N-methyl-1H-pyrazole-4-carboxamide (4f). A white powder;
yield 52.1%; M.p. 233–235°C; IR (KBr) ν: 3321, 3292,
1705, 1616, 1593, 1577, 1485, 1458, 1400, 1186, 1138,
1080, 1014, 970, 954, 867, 794, 761, 636. 1H NMR
(500 MHz, DMSO-d63) δ: 10.33(s, 1H, NH), 8.07(s,
1H, Py-H), 7.57(s, 1H, NH), 7.48(d, J = 8.0 Hz, 1H,
Ph-H), 7.32–7.29(m, 5H, Ph-H), 7.18(t, J = 8.0 Hz,1H,
1-(4-chlorophenyl)-5-(2-(2-chlorophenyl)-2-hydroxyacetamido)-
N-methyl-1H-pyrazole-4-carboxamide (4j).
A
white
powder; yield 52.7%; M.p. 203–205°C; IR (KBr) ν:
3365, 3278, 1718, 1624, 1600, 1579, 1500, 1485,
1454, 1411, 1394, 1377, 1273, 1182, 1082, 1035, 966,
835, 752. 1H NMR (500 MHz, DMSO-d6) δ: 10.35(s,
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2017
Synthesis and Biological Activity of Amide Compounds Containing Pyrazole
Mandelic Acid Groups
1H, NH), 8.12(s, 2H, Py-H), 7.46(d, J = 8.6 Hz, 4H, Ph-
H), 7.39(d, J = 3.5 Hz, 2H, Ph-H), 7.30(d, J = 8.6 Hz,
2H, Ph-H), 6.91(s, 1H, OH), 5.39(s, 1H, CH), 2.75(s,
3H, NCH3); 13C NMR (125 MHz, DMSO-d6) δ: 171.7,
162.4, 139.9, 130.1, 129.8, 129.7, 129.5, 127.6, 125.8,
112.7, 70.9, 40.2, 26.1. Anal. Calcd. for
C19H16ClN4O3: C 54.43, H 3.85, N 13.36; Found C
54.29, H 3.73, N 13.25
7.89(s, 1H, NH), 7.40–7.14 (m, 9H, Ph-H), 6.73–6.62(d,
J = 5.2 Hz, 1H, OH), 5.15–4.84(m, J = 4.6 Hz, 1H, CH),
2.76–2.59(d, J = 4.6 Hz, 3H, NCH3); 13C NMR
(125 MHz, DMSO-d6)) δ: 172.3, 162.3, 139.6, 139.5,
139.0, 136.6, 132.8, 129.1, 129.1, 128.5, 128.1, 123.8,
112.7, 73.3, 26.0. Anal. Calcd. for C19H17ClN4O3: C
59.30, H 4.45, N 14.56; Found C 59.02, H 4.56, N 14.70.
1-phenyl-5-(2-(2-fluorophenyl)-2-hydroxyacetamido)-N-methyl-
1H-pyrazole-4-carboxamide (4o). A white powder; yield
1-(4-chlorophenyl)-5-(2-hydroxy-2-phenylacetamido)-N-methyl-
1H-pyrazole-4-carboxamide (4k).
A white powder; yield
49.1%; M.p. 213–215°C; IR (KBr) ν: 3363, 3304, 3269,
1
48.2%; M.p. 83–85°C; H NMR (500 MHz, DMSO-d6) δ:
10.30(s, 1H, NH), 8.02(s, 1H, Py-H), 7.95(d, J = 4.6 Hz,
1H, NH), 7.34–7.22(m, 9H, Ph-H), 6.63(s, 1H, OH), 4.98(s,
1H, CH), 2.67(d, J = 4.6 Hz, 3H, NCH3); 13C NMR
(125 MHz, DMSO-d6) δ: 172.3, 162.4, 140.4, 139.8, 138.0,
136.8, 132.4, 129.3, 128.5,128.2, 127.3, 125.3, 112.5, 74.1,
26.1. Anal. Calcd. for C19H17ClN4O3: C, 59.30; H, 4.45; N,
14.56 ; Found C, 59.09; H, 4.28; N,14.32.
1716, 1624, 1600, 1587, 1487, 2462, 1448, 1396, 1276,
1
1068, 966, 756, 694. H NMR (500 MHz, DMSO-d6) δ:
10.27(s, 1H, NH), 8.04(d, J = 3.2 Hz , 1H, Py-H),
7.92(s, 1H, NH), 7.40(d, J = 6.5 Hz, 2H, Ph-H),
7.33(m, J = 18.6, 10.7, 6.1 Hz 5H, Ph-H), 7.11(dd,
J = 12.5, 5.2 Hz, 2H, Ph-H), 6.78(d, J = 5.2 Hz, 1H,
OH), 5.24(s, 1H, CH), 2.72(s, 3H, NCH3); 13C NMR
(125 MHz, DMSO-d6) δ: 171.6, 162.4, 139.5, 139.2,
136.6, 130.5, 129.7, 129.4, 128.2, 124.8,123.9,120.0,
115.7, 112.7, 68.0, 26.1, 19.8. Anal. Calcd. for
C19H17FN4O3: C 61.95, H 4.65, N 15.21; Found C
61.89,H 47,N 15.47.
1-(4-chlorophenyl)-5-(2-(4-chlorophenyl)-2-hydroxyxacetamido)-
N-methyl-1H-pyrazole-4-carboxamide (4l). A Pink crystal;
yield 53.2%; M.p. 83–85°C; IR (KBr) ν: 3361, 3269,
1707, 1625, 1581, 1500, 1485, 1456, 1411, 1394, 1377,
1
1313, 1273, 1188, 1089, 1060, 966, 831, 717, 692. H
1-phenyl-5-(2-hydroxy-2-phenylacetamido)-N-methyl-1H-
NMR (500 MHz, DMSO-d6) δ: 10.24(s, 1H, NH),
8.07(s, 1H, Py-H), 7.99(d, J = 4.6 Hz, 1H, NH), 8.09–
7.91(q, J = 8.6, 8.6 Hz, 4H, Ph-H), 7.28–7.22(q, J = 9.2,
9.2 Hz, 4H, Ph-H), 6.73(d, J = 4.6 Hz, 1H, OH), 5.03(d,
J = 4.6 Hz, 1H, CH), 2.73(d, J = 4.6 Hz, 3H, NCH3);
13C NMR (125 MHz, DMSO-d6) δ: 172.0, 162.3, 139.7,
139.3, 137.8, 136.8, 132.9, 132.6, 129.3, 129.0, 128.5,
125.4, 112.7, 73.2, 26.1. IR (KBr) ν: 3363, 3269, 1714,
1622, 1600, 1579, 1456, 1396, 1273, 1091, 1014, 833,
758. Anal. Calcd. for C19H16Cl2N4O3: C 54.43, H 3.85,
N 13.36; Found C 54.32, H 3.61, N,13.88.
pyrazole-4-carboxamide (4p).
A white powder; yield
50.8%; M.p. 87–88°C; IR (KBr) ν: 3360, 3305, 3273,
1705, 1624, 1600, 1587, 1489, 1462, 1448, 1375, 1276,
1186, 1149, 1060, 754, 721, 692.1H NMR (500 MHz,
DMSO-d6) δ: 10.30(s, 1H, NH), 8.04(s, 1H, Py-H),
7.97(d, J = 4.6 Hz, 1H, NH), 8.00–7.89(m,1H, CH),
7.36–7.16(m, 10H, Ph-H), 6.63(s, 1H, OH), 4.98(s, 1H,
CH), 2.68(d, J = 4.6 Hz, 3H, NCH3); 13C NMR
(125 MHz, DMSO-d6) δ: 172.5, 162.5, 140.5, 139.7,
139.1, 136.5, 129.3, 128.5, 128.2, 127.4, 123.6, 112.8,
74.1, 26.1, 24.8. Anal. Calcd. for C19H18N4O3: C 65.13,
H 5.18, N 15.99; Found C 64.87, H 5.51, N 5.60.
1-(4-chlorophenyl)-5-(2-(2-fluorophenyl)-2-hydroxyacetamido)-
N-methyl-1H-pyrazole-4-carboxamide (4m). A yellow crystal;
Antiviral bioassay against tobacco mosaic virus.
yield 50.1%; M.p. 229–231°C; IR (KBr) ν: 3361, 3267,
Purification of tobacco mosaic virus.
Using Gooding’s
1714, 1627, 1600, 1581, 1500, 1487, 1456, 1411, 1396,
1377, 1273, 1230, 1091, 1068, 968, 835, 754. H NMR
method [18], TMV was propagated, maintained in
Nicotiana tabacum cv.K326 and purified; the
concentration of virus was determined by an ultraviolet
spectrophotometer at 260 nm:
1
(500 MHz, DMSO-d6)) δ: 10.33(s, 1H, NH), 8.07(s, 1H,
Py-H), 7.99(s, 1H, Ph-H), 7.42–7.32(m, 6H, Ph-H), 7.12(q,
J = 8.9 Hz, 2H, Ph-H), 6.79(s, 1H, OH), 5.22(d,
J = 4.6 Hz, 1H, CH), 2.72(d, J = 4.6 Hz, 3H, NCH3); 13C
NMR (125 MHz, DMSO-d6)) δ: 171.7, 162.4, 139.8, 138.1,
136.9, 132.6, 130.4, 129.7, 129.4, 127.9, 125.6, 124.7,
115.9, 115.7, 112.5, 68.0, 26.1. Anal. Calcd. for
C19H16ClFN4O3: C 56.65, H 4.00, N 13.91; Found C
56.19, H 3.91, N 14.17.
virus concn ¼ ðA260ꢁdiluton ratioÞ=E0:1%;260nm
1cm
Curative effects of the compounds against tobacco mosaic
virus in vivo [19]. Growing leaves of N. tabacum L. of
the same age were selected. TMV (concentration
6 × 10ꢀ3 g · Lꢀ1) was dipped and inoculated onto
whole leaves that had been previously scattered with
siliconcarbide with a brush. The leaves were washed
with water and dried after inoculation. The compound
solution was smeared on the left side of the leaf, and
the solvent was smeared on the right side as a control.
Local lesion numbers were counted and recorded
1-phenyl-5-(2-(4-chlorophenyl)-2-hydroxyacetamido)-N-methyl-
1H-pyrazole-4-carboxamide (4n). A white powder; yield
56.3%; M.p. 83–85°C; IR (KBr) ν:3375, 3263, 1701,
1616, 1579, 1558, 1506, 1456, 1398, 1276, 1138, 1076,
1014, 954, 848, 763, 694. 1H NMR (500 MHz,
DMSO-d6)) δ: 10.22(s, 1H, NH), 8.08 (s, 1H, Py-H),
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Y. Xie, X.-H. Ruan, H.-Y. Gong, Y.-H. Wang, X.-B. Wang, J.-P. Zhang, Q. Li, and W. Xue
Vol 000
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3–4 days after inoculation. For each compound, three
replicates were conducted.
Protective effects of the compounds against tobacco mosaic
virus in vivo [19]. The compound solution was smeared
on the left side, and the solvent served as the control on
the right side of growing N. tabacum L. leaves of the
same age. The leaves that had been previously scattered
with
silicon
carbide
were
inoculated
with
6 × 10ꢀ3 g · Lꢀ1 of TMV after 12 h and then washed
with water. About 3–4 days after inoculation, the number
of local lesions was counted. Three replicates for each
compound were performed.
Inactivation effects of the compounds against tobacco mosaic
virus in vivo [19]. Tobacco mosaic virus was inhibited by
[8] Hu, D. Y.; Wan, Q. Q.; Yang, S.; Song, B. A.; Bhadury, P. S.;
Jin, L. H.; Yan, K.; Liu, F.; Chen, Z.; Xue, W. J Agric Food Chem 2008,
56, 998.
mixing with compound solution of the same volume for
30 min. The mixture was then inoculated on the left side
of the leaves of N. tabacumL, and the right side of the
leaves was inoculated with solvent and virus mixture as
control. The local lesion numbers were recorded
3–4 days after inoculation. Three repetitions were
conducted for each compound.
[9] Yakub, A.; Mohammad, S. A.; Hinna, H.; Asif, H.; Abhijeet,
D.; Sameena, B.; Chetna, K.; Syed, N.; Saqlain, H. J Med Chem Res
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Acknowledgments. We gratefully acknowledge the support by
the National Nature Science Foundation of China (No.
21462012), the Special Fund for Outstanding Scientific and
Technological Candidates of Guizhou Province (No. 201535).
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Journal of Heterocyclic Chemistry
DOI 10.1002/jhet