Novel pyrrolidine-2,4-dione derivatives containing pharmacophores of both hydrazine and diphenyl ether as potential antifungal agents: Design, synthesis, biological evaluation, and 3D-QSAR study

Article abstract of DOI:10.1039/d0nj04551a

In order to screen novel fungicide leads, 19 pyrrolidine-2,4-dione derivatives containing the pharmacophores of both hydrazine and diphenyl ether were designed, synthesized, and characterized by FT-IR, 1H NMR, 13C NMR, and HR-MS. The obtained target compo

Full text of DOI:10.1039/d0nj04551a

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Novel pyrrolidine-2,4-dione derivatives containing both
pharmacophores of hydrazine and diphenyl ether as potential
antifungal agents: design, synthesis, biological evaluation, and
Received 00th January 20xx,
Accepted 00th January 20xx
3D-QSAR study†
DOI: 10.1039/x0xx00000x
a
a
a
a
ab
ab
Hao-Ran Hu, An Wang, Ling-Ling Qiu, Xiao-Bin Wang, Min Chen, Ai-Min Lu and Chun-Long
Yang*
www.rsc.org/
ab
In order to screen novel fungicide leads, nineteen pyrrolidine-2,4-dione derivatives containing both pharmacophores of
hydrazine and diphenyl ether were designed, synthesized, and characterized by FT-IR, H NMR, 13C NMR, and HR-MS
spectra. The obtained target compounds were evaluated for their antifungal activity aganist Rhizoctonia solani (Rs), Botrytis
cinerea (Bc), and Fusarium graminearum (Fg). The bioassay results indicated that the majority of target compounds
exhibited striking biological activities against the mentioned-above fungi. Strikingly, the target compound 4h obviously
inhibited the growth of Rs in vitro with the EC50 value of 0.39 μg/mL, which was better than the commercialized fungicide
boscalid (2.21 μg/mL). Subsequently, the compound 4h was further evaluated for the in vivo anti-Rs effect on rice leaves
and presented the control efficiency of 45.33% at 200 μg/mL. The scanning electron microscopy (SEM) was taken for
observing the changes on mycelium morphology. A lot of abnormal branches and swellings were found on the hyphae that
were treated with the compound 4h. Based on the bioassay results against Rs, the comparative molecular similarity index
analysis model was built to further analyze the structure-activity relationship of target compounds. The above 3D-QSAR
1
2
analyses revealed that introducing hydrophobic 4-FPh, 4-ClPh, or 4-BrPh fragment into the R position could greatly
increase the antifungal activity against Rs.
Introduction
The food increases on qualities and quantities notoriously bring
numerous ecnomic and social benifits for human development.
However, the plant fungi including Rhizoctonia solani (Rs), Botrytis
cinerea (Bc), and Fusarium graminearum (Fg) threaten the
Tetramic acids, the significant natural products widely existing
in marine and terrestrial organisms, were notoriously reported for
their favourable compatibility with the environment.
1
11
Remarkably, tetramic acids were also documented for their wide
2–5
12
productivity and quality of agricultural econimic crops.
Fortunately, the agricultural fungicides can help protect crops,
bioactivities against agricultural pests, such as antifungal,
13
14
15
antibacterial,
activities.
herbicidal,
insecticidal,
and antiviral
6
7
8
16,17
such as rice, strawberry, and wheat, from the harm that was
caused by the above phytopathogenic fungi. While, a large
number of plant fungi have developed the resistances against
Among them, tetramic acids containing
a
pyrrolidine-2,4-dione scaffold have attracted numerous attentions
due to their potential utilization as antifungal leads. For example,
9
18
19
conventional fungicides over the past decades. The serious
cryptocin and harzianic acid (Fig. 1)
were successively
consequences that threaten the survival of mankind will be caused
if researchers do not pay attentions to the mentioned-above
problem. Therefore, it is an imminent matter to develop neoteric
separated as the naturally antifungal tetramic acids bearing a
pyrrolidine-2,4-dione scaffold. Meanwhile, in our previous work, a
20
21
22
23
substituted amine, pyrrole, phenylhydrazine, or strobilurin
10
agricultural fungicides.
moiety was introduced into pyrrolidine-2,4-dione framework to
successfully construct some novel tetramic acid derivatives that
exhibited
the
satisfactory
inhibition
effect
against
phytopathogenic fungi (Fig. 1).
a
Jiangsu Key Laboratory of Pesticide Science, Nanjing Agricultural University,
Nanjing, 210095, PR China.
Department of Chemistry, College of Sciences, Nanjing Agricultural University,
Nanjing, 210095, PR China.
Hydrazine fragments have attached prevalent attentions from
agricultural and medicinal fields due to their specific features
including simple molecular structures, convenient preparation
b
*
Corresponding author. E-mail address: ycl@njau.edu.cn (C.L. Yang)
†
Electronic supplementary information (ESI) available: Copies of FT-IR, 1 H NMR,
C NMR, and HRMS spectrograms for all title compounds can be found. For ESI or
methods, multiple bioactivities, and low toxicity against
1
3
24–26
mammals.
The fungicides containing a hydrazine, such as
other electronic format see DOI:
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Fig. 1 Design strategy of target compounds
.
famoxadone and benquinox, were widely applied in agricultural
attentions. We made bold a conjecture based on the above points,
that is, introducing diphenyl ether fragment into the
27,28
production.
In our previous work,
a
series of
a
pyrrolidine-2,4-dione derivatives containing N-substituted
phenylhydrazine moiety were constructed and reported for their
obvious antifungal activities against phytopathogenic fungi.
Unfortunately, the in vivo antifungal effects of the
mentioned-above pyrrolidine-2,4-dione derivatives were not
satisfactory, which might be caused by their poor lipid solubility
within living organisms. Therefore, improving the lipid solubility of
pyrrolidine-2,4-dione derivatives has been focused as the key
points that we’re going to work on.
pyrrolidine-2,4-dione nuclei containing a phenylhydrazine moiety
might be served as a good solution for improving the in vivo
antifungal effects of bioactive molecules. Subsequently, our
research work was continued by introducing a lipophilic diphenyl
ether fragment into the 1-position of the pyrrolidine-2,4-dione
nucleus that was constructed in our previous work (Fig. 1).
Nineteen novel pyrrolidine-2,4-dione derivatives containing
hydrazine and diphenyl ether pharmacophores were designed and
synthesied to explore their antifungal activity against
phytopathogenic fungi. Afterwards, a comparative molecular
similarity index analysis (CoMSIA) was constructed to analyze the
effects of steric, electrostatic, hydrophpbic and H-bond fieldson on
the antifungal activities of target compounds.
2
2
As the bioactive lipophilic substructures existing in the
commercial agricultural fungicides including aminopyrifen (Fig.
), mefentrifluconazole (Fig. 1),30 and difenoconazole, the
2
9
31
1
diphenyl ether pharmacophores deserved our numerous
Scheme 1 Synthetic route to target compounds 4a–4s. Reagaents and conditions: (a) CH
3
COONa (1.2 equiv), BrCH
2
COOC
2
H
5
(1.2 equiv), C
2
5
H OH, reflux; (b)
Diketene (1.0 equiv), r.t.; (c) C
2
H
5
ONa (1.0equia), CH
3
OH, reflux; (d) 10% HCl, acidification to pH=2; (e) C
2
H
5
OH, reflux.
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View Article Online
DOI: 10.1039/D0NJ04551A
Results and discussion
Synthesis of target compounds
The synthetic route to pyrrolidine-2,4-dione derivatives containing
hydrazine and diphenyl ether pharmacophores 4a–4s is shown in
Scheme 1. The substitution reaction of
-phenoxyaniline with ethyl bromoacetate took place in the
refluxed anhydrous ethanol containing sodium acetate to
a
substituted
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generate an ethyl (4-(substituted phenoxyphenyl))glycinate 1.
Next, diketene was added to the ethanol containing the
intermediate 1, and the resulting mixture was stirred at room
temperature to obtain an ethyl N-(3-oxobutanoyl)-N-(4-
Fig. 2 Tautomeric equilibria of target compounds.
33
(
substituted phenoxyphenyl))glycinate 2. After that, the Claisen
condensation inside the molecule of intermediate 2 was activated
by sodium ethoxide in the refluxed methanol. And the obtained
mixture was acidify until pH reached
-(1-hydroxy-ethylidene)-1-(4-(substituted
2
to obtain the
phenoxyphenyl))-
isomers A with Z-configuration and C with E-configuration are
absolutely stable relative to the corresponding enol isomers B and
D according to the crystallographic analysis and computational
3
pyrrolidine-2,4-dione 3. Finally, the reaction of an intermediate 3
with a substituted hydrazide took place in refluxed ethanol to
generate the target compound 4. Using the similar methods, the
target compounds 4a–4s were successively synthesized in yields
ranging from 21.5% to 65.2%.
3
4,37
chemistry.
The compound 4h was used as an example to
3
4
calculate the relative energy of its four different isomers by
SYBYL-X 2.0 software (Shanghai. Tri-I. biotech. Inc., China). The
results showed that the relative energy of A, B, C, and D isomers
were 1.112, 8.143, 0.000, and 4.031 kJ/mol, respectively.
Obviously, the keto isomers A and C caused the characteristic pair
of signals of NMR. The E-isomer of C should be the dominant form,
Spectral characteristics of target compounds
The spectral characteristics of target compound 4h were analyzed
as a typical example that was described below. In the IR spectrum
of target compound 4h, the absorption peaks at 3413 and 3234
1
which is more stable than the Z-isomer A. As shown in the H NMR
spectrum, the CONH proton of compound 4h gave two singlets at
1.65 ppm and 11.91 ppm in a ratio of 64: 36, which represented
1
–1
cm were caused by the stretching vibration of the N-H bonds on
the respective contents of E- and Z-isomers.
–1
hydrazine group. The obvious peaks at 1632 and 1685 cm were
contributed to the carbonyl at the 2- and 4-positions of
pyrrolidine-2,4-dione fragment. The peak at 1232 cm meant the
In vitro antifungal activity of target compounds
–
1
1
existence of ether bond. The H NMR spectrum of compound 4h
A mycelium growth method was used to evaluate the biological
activities of the target compounds against three
phytopathogenic fungi (Rs, Fg, and Bs), with the commercial
showed the two singlets at 11.65 and 5.93 ppm, which confirmed
that the target compound 4h has a CONHNH fragment in its
molecular structure. The singlet at 4.14 ppm was attributed to the
3
8–40
fungicides carbendazim and boscalid as the positive controls.
protons of the CH
2
fragment at the 5-position of
As shown in Table 1, the target compounds 4d and 4g exhibited
obvious anti-Rs effect at 50 μg/mL, with the corresponding
inhibition rates of 91.7% and 88.2 %, which were better than that
of boscalid (87.5%). Meanwhile, Table 1 also showed that the
inhibition rates of target compounds 4a, 4b, and 4l against Bc at
50 μg/mL were respectively 69.2%, 64.8%, and 66.1%. To further
research the inhibition effect of target compounds against Rs, all
compounds were tested for their anti-Rs EC50 values. Similar to the
antifungal bioassay results mentioned-above, Table 2 showed that
the anti-Rs EC50 values of the compounds 4d, 4e, 4h, 4j, 4k, 4p,
and 4r were 1.31, 1.75, 0.39, 0.50, 2.14, 1.01, and 0.97 μg/mL,
respectively, which were obviously better than that of boscalid
(2.21 μg/mL). Among them, the in vitro anti-Rs effects of
compound 4h was actually shown in Fig. 3.
1
3
pyrrolidine-2,4-dione core. In the C NMR spectrum, two singlets
at 191.06 and 171.89 ppm indicated the existence of two carbonyl
groups in compound 4h. At the same time, the two singlets at
56.10 and 13.44 ppm also confirmed the presences of CH
2
and CH
3
+
groups in the 4h. In the HR-MS spectrum, the [M + H] ion peak of
target compound 4h was shown as m/z 434.1263 that matches
with the theoretical value.
The structures of the target compounds were interesting since
solubilized tetramic acids and their derivatives commonly display
3
5,36
four tautomeric forms.
The pyrrolidine-2,4-dione derivatives
containing hydrazine and diphenyl pharmacophores 4a–4s possibly
existed in four tautomeric forms A, B, C, and D (Fig. 2). The
internal tautomers A⇌B and C⇌D were rapidly interconverted by
the intramolecular displacement of the enolic proton along with
the hydrogen bond, whereas the interconversion between
external tautomers AB⇌CD is a comparatively slow process in the
In vivo antifungal activity of compound 4h
Considering the obvious inhibitory effects of target compounds
aganist Rs, the compound 4h with the minimum EC50 value of 0.39
μg/mL was selected as a representative to evaluate the in vivo
control effect against Rs on rice leaves. As seen from Fig. 4, the
in vivo anti-Rs control efficiency of 4h reached 45.33% at 200
3
5,36
NMR timescale.
Therefore, the external tautomers gave the
1
13
separate NMR signals in both H NMR and C NMR spectra of
target compounds. And the H NMR signals of affected protons
split in a ratio of 60–68: 32–40. In the internal tautomers, the keto
1
41
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Table 1 Antifungal activities of target compounds against Rs, Bc, and Fg
View Article Online
DOI: 10.1039/D0NJ04551A
a
Inhibitory rate at 50 μg/mL （％）
Compd.
R1
R2
Rs
Bc
Fg
4
4
4
a
b
c
H
H
H
H
CH
Ph
2-FPh
4-FPh
2-ClPh
3-ClPh
4-ClPh
2,4-diClPh
4-BrPh
4-CF Ph
CH
Ph
40.6 ± 0.2
60.1 ± 0.9
81.0 ± 0.2
91.7 ± 0.3
84.7 ± 0.3
78.8 ± 0.1
88.2 ± 0.4
80.9 ± 0.6
58.5 ± 0.3
76.4 ± 0.9
84.1 ± 0.2
60.1 ± 0.1
75.3 ± 0.3
82.8 ± 0.2
77.0 ± 0.9
80.8 ± 1.0
74.3 ± 0.7
80.5 ± 1.5
75.2 ± 1.5
69.2 ± 1.4
64.8 ± 0.4
38.2 ± 0.4
24.6 ± 0.1
55.5 ± 0.2
16.0 ± 0.4
37.8 ± 0.4
50.1 ± 0.4
42.0 ± 0.3
34.9 ± 0.4
39.4 ± 0.6
66.1 ± 0.2
20.9 ± 0.3
16.2 ± 0.2
42.8 ± 0.3
34.2 ± 0.5
18.1 ± 0.5
25.8 ± 0.4
23.3 ± 0.3
36.0 ± 1.4
47.2 ± 0.6
31.2 ± 0.5
14.3 ± 0.5
59.8 ± 0.3
13.7 ± 0.2
18.2 ± 0.2
48.9 ± 0.2
35.2 ± 0.1
42.4 ± 0.3
22.3 ± 0.4
50.8 ± 0.3
18.4 ± 0.2
20.7 ± 0.4
34.1 ± 0.3
31.9 ± 0.4
19.8 ± 0.7
20.3 ± 0.4
13.9 ± 0.2
100.0± 0.0
—
3
4d
4e
4f
H
H
H
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0
4
4
4
4
4
4
4
4
4
4p
4q
4r
4
Car
Bosb
g
h
i
j
k
l
m
n
o
H
H
H
H
H
3
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
—
—
3
Fig. 3 In vitro anti-Rs effect photographs of compound 4h. A: 4h at 5
µg/mL; B: 4h at 2.5 µg/mL; C: 4h at 1.25 µg/mL; D: 4h at 0.625 µg/mL; E: 4h at
0.3125 µg/mL; F: blank control.
2-FPh
4-FPh
4-ClPh
2,4-diClPh
4-BrPh
s
4-CF
—
3
Ph
b
100.0 ± 0.0 100.0 ± 0.0
87.5 ± 0.3 98.3 ± 0.0
—
20%
40%
60%
80%
Inhibitory Effect
a
Average of three replicates. b Commercial agricultural fungicides
carbendazim (Cab), and boscalid (Bos) were used for comparison of
antifunal activity.
Fig. 4 In vivo control efficiency photographs of compound 4h against Rs
on rice leaves at 200 μg/mL. A: 4h at 200 μg/mL; B: hymexazol at 200 μg/mL;
C: boscalid at 200 μg/mL; D: carbendazim at 200 μg/mL; E: blank control.
a
Date were obtained from five parallel experiments.
Table 2 EC50 and pEC50 values of target compounds 4a–4s against Rs
Compd.
EC50a
pEC50
Predictedb
Residualc
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
a
b
c
26.72 ± 1.31
19.92 ± 1.48
2.54 ± 0.01
1.31 ± 0.01
1.75 ±0.02
13.82 ± 0.97
2.47 ± 0.04
0.39 ± 0.01
7.76 ± 0.29
0.50 ± 0.01
2.14 ± 0.01
11.46 ± 0.32
7.78 ± 0.10
25.17 ± 0.63
2.29 ± 0.37
1.01 ± 0.04
11.24 ± 0.48
0.97 ± 0.01
7.33 ± 0.01
0.34 ± 0.01
2.21 ± 0.04
4.080
4.230
5.200
5.500
5.380
4.500
5.240
6.050
4.780
5.980
5.340
4.510
4.750
4.250
5.300
5.670
4.650
5.720
4.840
4.080
4.551
5.295
5.003
5.494
4.393
5.247
5.868
4.966
6.109
5.264
4.193
4.938
4.646
5.137
5.511
4.609
5.752
4.904
0.000
–0.321
–0.095
0.497
–0.114
0.107
–0.007
0.182
–0.216
–0.129
0.076
μg/mL, which was better than that of hymexazol (35.64%) under
the same conditions. Unfortunately, this in vivo anti-Rs activity
mentioned-above was inferior to that of boscalid (76.48%) and
carbendazim (79.46%) at 200 μg/mL. The in vivo bioassay results
showed a significant guidance for the further molecular
optimization of novel pyrrolidine-2,4-dione derivatives.
d
e
f
g
h
i
Structrue-activity relationships
j
Based on the bioassay results in Table 1 and 2, introducing
k
l
1
2
disparate substituents at the R and R positions remarkably
influenced the antifungal activity of the target compounds. Some
structure-activity relationships are summarized as follows. Firstly,
all target compounds except 4a, 4b, and 4l exhibited better
antifungal activity against Rs than that against Bc and Fg.
Secondly, the compounds 4a, 4b, and 4l bearing a hydrogen atom
or methyl fragment at the R position displayed better anti-Bc
activities than the other compounds bearing a phenyl (4c–4k) or
substituted phenyl group (4m–4s) at the R position. Thirdly, the
bioassay results also revealed that the presence of the 4-FPh,
0.317
m
n
o
p
q
r
–0.188
–0.396
0.163
0.159
0.041
–0.032
–0.064
2
s
2
Card
Bosd
—
—
—
—
—
—
2
4-ClPh, or 4-BrPh group at the R position could enhance the
a
Average of five replicates. b Predicted pEC50 by CoMSIA. c Relative
anti-Rs effect of target compounds. For example, the anti-Rs EC50
d
error of experimentally predicted pEC50
carbendazim (Car), and boscalid (Bos) were used as positive controls.
.
Agricultural fungcides
1
2
1
2
values of compounds 4e (R = H, R = 4-FPh), 4h (R = H, R =
1
2
1
2
1
4-ClPh), 4p (R = Cl, R = 4-ClPh), 4j (R = H, R = 4-BrPh), and 4r (R
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2
=
Cl, R = 4-BrPh) reached 1.75, 0.39, 1.01, 0.50, and 0.97 μg/mL,
View Article Online
DOI: 10.1039/D0NJ04551A
which are better than other target compounds. Lastly, introducing
a chlorine atom at the R position did not significantly enhance the
antifungal effect of target compounds against Rs, Bc, and Fg. For
example, the compounds 4n (R = Cl, R = 2-FPh), 4p (R = Cl, R =
1
1
2
1
2
1
2
4-ClPh), and 4s (R = Cl, R = 4-CF3Ph) all showed lower antifungal
Fig. 5 Alignment of all target compunds of 3D-QSAR study. A: template
molecular structure 4h; B: result of alignment with common scaffold
marked in red circles.
1
2
activity than the corresponding compounds 4d (R = H, R = 2-FPh),
1
2
1
2
4
h (R = H, R = 4-ClPh), and 4k (R = H, R = 4-CF
3
Ph).
0
1
2
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9
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Study of 3D-QSAR model
Table 3 Statistical parameters for CoMSIA model
According to the experimental pEC50 values (Table 2), the CoMSIA
model was constructed to further analyze the structure-activity
relationship (SAR) of nineteen target compounds against Rs. Based
on the common scaffold that was marked with red circles (Fig. 5B)
on the template molecule 4h (Fig. 5A), three-dimensional
structures of compounds 4a–4s were aligned, and the obtained
results were showed in Fig. 5B. The corresponding statistics were
calculated and listed in Table 3. The internal validation of
cross-validated coefficient (q ) and the non-cross-validated
correlation coefficient (r ) indicated the credible level of the
constructed 3D-QSAR model, and the values q > 0.5 and r > 0.8
are normally regarded as the necessary requirements for the
Statistical parameter
CoMSIA
Validation criteria
>0.5
q2a
ONCb
0.607
6
r2c
SEEd
0.872
0.262
13.581
>0.8
F-test value
Field contributions
Steric
0.132
0.107
0.689
0
2
Electrostatic
Hydrophobi
H-bond donor
H-bond acceptor
2
2
2
0.071
38
reliable 3D-QSAR model. As shown in Table 3, the CoMSIA model
a
c
Cross-validated correlation. b Optimum number of components.
Non-cross-validated correlation. d Standard error of estimate.
2
2
2
2
had the r of 0.872 (r > 0.8) with 6 ONC, the q of 0.607 (q > 0.5),
the F test value of 13.581, and the SEE of 0.262. The above
statistic characteristic data indicated that the obtained CoMSIA
model has a high predictive accuracy.
In the CoMSIA model, the contributions of steric, electrostatic,
hydrophobic, H-acceptor, and H-bond donor fields are respectively
0.132, 0.107, 0.689, 0.071, and 0.000, which meant clearly that
the hydrophobic field played the most significant role in
maintaining the biological activity against Rs. Afterwards, the
corresponding pEC50 values of target compounds against Rs were
predicted by the obtained CoMSIA model, and the results are
shown in Table 2. The correlations between the experimental and
predicted pEC50 values against Rs (Fig. 6) further confirmed the
reliability of the obtained CoMSIA model.
The CoMSIA profile maps including steric, electrostatic,
hydrophobic, and H-bond acceptor fields were presented in Fig. 7.
In the steric field that was showed in Fig. 7A, the large groups in
green module could increase the anti-Rs activity, while their
presences in yellow contours would decrease the anti-Rs activity.
As shown in Fig. 7A, the bulky green profile near the phenyl ring of
phenylhydrzine scaffold explained that the bulky substituents at
Fig. 6 Correlation between experimental and predicted pEC50 values in
CoMSIA model.
2
the R position could improve the anti-Rs effects. This observation
clarified the facts that the target compounds bearing a phenyl ring
2
7B explained that introducing electron-donating groups into the
blue region near the 3-position in the phenyl ring of
phenylhydrazine fragments would be advantageous for anti-Rs
activities. What’s more, the red contours near the 2- and
at the R position had the higher biological activity than the
compounds bearing a hydrogen atom or methyl fragment at the
same position. On the other hand, the little yellow profile around
the 4-position of the phenyl ring at phenylhydrazine scaffolds
suggested that introducing bulky substituents into this region was
unfavorable for the improvement of biological activity. In the
profile map of the CoMSIA electrostatic fields (Fig. 7B), introducing
electron-donating fragments into blue regions would increase
anti-Rs activity, and introducing electron-withdrawing groups in
red profiles would also increase the inhibition rates against Rs. Fig.
1
4-positions at phenyl ring of phenylhydrazine fragments and the R
position at diphenyl ether fragment suggested that introducing
appropriate electron-withdrawing substitutes into these regions
would enhance anti-Rs activities. In the profile maps of CoMSIA
hydrophobic field, the grey and yellow contours indicated that the
hydrophobic and hydrophilic substitutes could increase the
inhibition rates against Rs. As shown in Fig. 7C, the yellow region
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DOI: 10.1039/D0NJ04551A
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Fig. 7 CoMSIA profile maps with compound 4h shown inside fields. A
steric fields): large groups in the green and yellow regions could increase
Fig. 8 SEM photographs of hyphae from Rs colony. A, B, and C: blank
control of Rs; D, E, and F: Rs treated with compound 4h at 10 μg/mL.
(
or decrease the bioactivites of target compounds; B (electrostatic fields):
electron-donating and electron-withdrawing groups in the blue and red
regions could increase the bioactivity of target compounds; C (hydrophobic
fields): hydrophobic and hydrophilic groups in the yellow and gray regions
could increase the bioactivity of target compounds; D (hydrogen-bond
acceptor fields): H-bond acceptor groups in the purplish red regions could
increase the bioactivity of target compounds.
Japan). A thin-layer hromatography (TLC) was performed on silica
gel GF254 to monitor reactions under ultraviolet light 254 nm. High
resolution mass spectra (HRMS) were performed on a Finnigan
TRACE2000 gas chromatograph-mass spectrometer (JEOL, Japan).
All reagents and solvents are analytical or chemically pures.
near the 4-position at the phenyl ring of phenylhydrazine
fragments suggested that the hydrophobic substitutes are favored
there. The hydrophilic groups, such as fluorine, chloride, and
bromine atoms, should be introduced into the 4-position at the
phenyl ring of phenylhydrazine fragments. Finally, as shown in Fig.
Synthesis procedure for intermediates 3a and 3b
A substituted 4-phenoxyaniline (0.025 mol) and sodium acetate
0.03 mol) were stirred in refluxed ethanol (50 mL). After the
above reaction was completed, the resulting mixture was
processed by reduced pressure distillation, poured into cold water
50 mL), extracted with benzene, dried with anhydrous sodium
sulfate, and removed the solvent to prepare the intermediate 1.
Then, diketene (0.025mol) was slowly added into the anhydrous
ethanol (50 mL) containing the intermediate 1 (0.025 mol) in an
ice-salt bath, and the mixture was stirred at room temperature for
(
7D, the profile map in H-bond fields demonstrated that H-acceptor
(
groups in the purple regions would increase the bioactivity,
suggesting that the hydrazine fragment played a vital role in
improving the bioactivity of the target compounds against Rs.
SEM study on hyphae morphology
1
0 h to synthesize the intermediate 2. After that, the sodium
In order to investigate the effect of compound 4h on mycelium
morphology, the hyphae treated with compound 4h at 10 μg/mL
methoxide (0.025mol) dissolved in anhydrous methanol (10 mL)
was added in the mentioned-above mixture and stirred under
reflux for 5 h. After the redundant methanol was removed, the
obtained residue was dissolved in distilled water (10 mL), and then
extracted with diethyl ether (20 mL × 3). The water layer was
acidified with 10% hydrochloric acid until the pH value reached 1–
, and extracted with diethyl ether. The obtained extract was
washed with water, dried with anhydrous sodium sulfate, and
concentrated in vacuo to give the important intermediate 3a or
4
2
was studied by a scanning electron microscope (SEM). As shown
in Fig. 8A–8C, the mycelia growing in the blank control were
regular, and the mycelia were arranged in order with intact shapes
and rounded surfaces. On the contrary, after treated by
compound 4h, the drastic changes in mycelial morphologies were
observed in Fig. 8D–8F. The mycelial growth appeared unnormal,
the arrangement of mycelia became distorted, and the hyphae
showed numerous branches and swellings.
2
3b.
3
-(1-Hydroxyethylidene)-1-(4-phenoxyphenyl)pyrrolidine-2,4-
1
dione (3a). Yellow solid, yield 52.6%; H NMR(400 MHz, CDCl
3
) δ:
Experimental
General
E-isomer: 7.63–7.60 (m, 2H, ArH), 7.37 (t, J = 7.9 Hz, 2H, ArH), 7.14
t, J = 7.4 Hz, 1H, ArH), 7.08 (d, J = 9.1 Hz, 2H, ArH), 7.03 (d, J = 7.8
Hz, 2H, ArH), 4.32 (s, 1H, CH ), 2.62 (s, 3H, CH ); Z-isomer: 7.63–
.60 (m, 2H, ArH), 7.37 (t, J = 7.9 Hz, 2H, ArH), 7.14 (t, J = 7.4 Hz,
H, ArH), 7.08 (d, J = 9.1 Hz, 2H, ArH), 7.03 (d, J = 7.8 Hz, 2H, ArH),
.18 (s, 1H, CH ), 2.56 (s, 3H, CH ).
-(4-(4-Chlorophenoxy)phenyl)-3-(1-hydroxyethylidene)-
pyrrolidine-2,4-dione (3b). Yellow solid, yield 39.5%; H NMR(400
MHz, CDCl ) δ: E-isomer: 7.61 (dd, J = 8.9, 3.9 Hz, 2H, ArH), 7.31–
.28 (m, 2H, ArH), 7.06–7.02 (m, 2H, ArH), 6.94 (dd, J = 9.4, 2.6 Hz,
H, ArH), 4.29 (s, 2H, CH ), 2.60 (s, 3H, CH ); Z-isomer: 7.61 (dd, J =
(
Melting points (m.p.) of target compounds were determined by an
uncorrected SMP50 Automatic Melting point Apparatus
2
3
7
1
4
(
Staffordshire, United Kingdom). Infrared spectra (IR) of synthetic
compounds were recorded on a Thermo Nicolot 380 FTIR
spectrometer (Thermo Nicolet Corporation, America). Using
DMSO-d or CDCl as solvent, H NMR and C NMR spectra were
6 3
recorded on a BRUKER, AVANCE III 400 MHz (Bruker Corporation,
Bremen, Germany) at room temperature. The scanning electron
microscope (SEM) was observed on a Hitachi SU8018 (Hitachi,
2
3
1
1
1
13
3
7
2
2
3
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1
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1
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2
2
2
2
2
2
2
2
2
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6
8.9, 3.9 Hz, 2H, ArH), 7.31–7.28 (m, 2H, ArH), 7.06–7.02 (m, 2H,
2.71 (s, 3H, CH
3
); Z-isomer: 11.72 (s, 1H, NHN), 7.62 (d, J = 9.0 Hz,
View Article Online
DOI: 10.1039/D0NJ04551A
ArH), 6.94 (dd, J = 9.4, 2.6 Hz, 2H, ArH), 4.16 (s, 2H, CH
2
), 2.54 (s,
2H, ArH), 7.35–7.27 (m, 4H, ArH), 7.11–6.98 (m, 6H, ArH), 6.80 (d, J
3
H, CH
3
).
= 7.7 Hz, 2H, ArH), 5.91 (s, 1H, NHAr), 4.14 (s, 2H, CH
2
), 2.71 (s, 3H,
) δ: E-isomer; 191.05, 172.01,
171.24, 157.50, 153.23, 146.07, 134.32, 129.80, 129.63, 123.18,
22.08, 120.91, 119.72, 118.50, 113.28, 96.25, 56.04, 13.51;
Z-isomer: 193.90, 172.35, 168.92, 157.60, 153.00, 145.97, 134.93,
13
3 3
CH ); C NMR (101MHz, CDCl
Synthesis procedure for target compounds 4a-4s
1
The intermediate 3a (1 mmol) and phenylhydrazine (1.5 mmol)
were dissolved in ethanol (20 mL) and were stirred under reflux.
After the mixture was clear and transparent, the heating was
stopped and the reaction continued at room temperature for 2 h.
By standing precipitation, the produced solid was filtered from the
mixture, and washed several times with small amount of ethanol
to give the target compound 4c. Using similar method, other
target compounds 4a, 4b and 4d-4s were synthesized successively.
1
29.78, 129.63, 123.09, 122.16, 121.16, 119.72, 118.44, 113.24,
0
1
2
3
4
5
6
7
8
9
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8
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0
9
7.74, 54.05, 12.43.; HR-MS (ESI): m/z calcd for C24
H
22
N
O
3 3
([M +
+
H] ) 400.1655, found 400.2056.
3
-(1-(2-(2-Fluorophenyl)hydrazinyl)ethylidene)-1-(4-phenoxy-
phenyl)pyrrolidine-2,4-dione (4d). White soild, yield 62.1%, m.p.
75.4–176.2 ℃; IR (KBr, cm ) v: 3414, 3131, 1679, 1620, 1579,
503, 1488, 1399, 1299, 1196, 1157, 869, 778; H NMR(400 MHz,
–
1
1
1
1
3-(1-Hydrazinylethylidene)-1-(4-phenoxyphenyl)pyrrolidine-2,4
CDCl
.32 (m, 2H, ArH), 7.11–6.91 (m, 8H, ArH), 6.85 (t, J = 8.0 Hz, 1H,
ArH), 6.13 (s, 1H, NHAr), 4.15 (s, 2H, CH ), 2.73 (s, 3H, CH );
Z-isomer: 11.88 (s, 1H, NHN), 7.62 (d, J = 8.7 Hz, 2H, ArH), 7.32 (m,
H, ArH), 7.11-6.91 (m, 8H, ArH), 6.85 (t, J = 8.0 Hz, 1H, ArH), 6.13
3
) δ: E-isomer: 11.65 (s, 1H, NHN), 7.62 (d, J = 8.7 Hz, 2H, ArH),
-
(
dione (4a): Yellow solid, yield 58.2%, m.p. 196.0–196.8 ℃; IR
KBr, cm ) v: 3311, 3068, 1627, 1586, 1503, 1489, 1432, 1244,
7
–1
1
2
3
3
1209, 1163, 982, 919, 877, 744; H NMR(400 MHz, CDCl ) δ:
E-isomer: 11.55 (s, 1H, NHN), 7.60 (s, 2H, ArH), 7.32 (t, J = 7.9 Hz,
2
(
(
1
1
1
1
2
H, ArH), 7.08 (t, J = 7.3 Hz, 1H, ArH), 7.03 (s, 1H, ArH), 7.00 (d, J =
.4 Hz, 2H, ArH), 6.98 (s, 1H, ArH), 4.07 (s, 2H, CH ), 3.98 (s, 2H,
), 2.73 (s, 3H, CH ); Z-isomer: 11.96 (s, 1H, NHN), 7.60 (s, 2H,
ArH), 7.32 (t, J = 7.9 Hz, 2H, ArH), 7.08 (t, J = 7.3 Hz, 1H, ArH), 7.03
1
3
s, 1H, NHAr), 4.15 (s, 2H, CH
101MHz, CDCl ) δ: E-isomer: 191.02, 171.35, 171.87, 157.52,
52.21, 152.95, 134.28, 129.77, 125.01 (d, J = 3.6 Hz), 123.13,
2 3
), 2.73 (s, 3H, CH ); C NMR
5
NH
2
2
3
3
22.22, 121.04, 120.80, 119.72, 118.54, 115.49 (d, J = 17.7 Hz),
14.25 (d, J = 1.7 Hz), 96.63, 56.04, 13.43; Z-isomer: 194.08,
72.57, 168.61, 157.52, 152.26, 149.86, 134.16, 129.77, 125.01 (d,
J = 3.6 Hz), 123.04, 122.15, 121.04, 120.80, 119.72, 118.39, 115.53
d, J = 17.6 Hz), 114.19, 98.12, 53.99, 12.35; HR-MS (ESI): m/z calcd
([M + H]+) 418.1561, found 418.1566.
-(1-(2-(4-Fluorophenyl)hydrazinyl)ethylidene)-1-(4-phenoxy-
phenyl)pyrrolidine-2,4-dione (4e). Light yellow solid, yield 55.4%,
(
s, 1H, ArH), 7.00 (d, J = 5.4 Hz, 2H, ArH), 6.98 (s, 1H, ArH), 4.07 (s,
13
2H, CH
2
), 3.98 (s, 2H, NH
2 3
), 2.73 (s, 3H, CH ); C NMR (101MHz,
DMSO-d
6
) δ: E-isomer: 187.98, 169.22, 165.62, 157.98, 151.48,
1
1
1
36.33, 130.40, 123.33, 120.40, 119.99, 118.20, 100.21, 55.00,
2.96; Z-isomer: 187.98, 169.22, 165.62, 157.98, 151.48, 136.33,
30.40, 123.33, 120.40, 119.99, 118.20, 100.21, 55.00, 12.96;
(
3 3
for C24H21FN O
3
+
18 3 3
HR-MS (ESI): m/z calcd for C18H N O ([M + H] ) 324.1342, found
3
24.1341.
-(1-(2-Methylhydrazinyl)ethylidene)-1-(4-phenoxyphenyl)-
pyrrolidine-2,4-dione (4b). Yellow solid, yield 40.1%, m.p. 175.1–
–1
m.p. 179.4–180.8 ℃; IR (KBr, cm ) v: 3413, 3256, 3134, 1681,
3
1
1
625, 1582, 1498, 1488, 1469, 1399, 1376, 1227, 947, 870, 828; H
NMR(400 MHz, CDCl
.0 Hz, 2H, ArH), 7.32 (t, J = 7.9 Hz, 2H, ArH), 7.09 (t, J = 7.4 Hz, 1H,
ArH), 7.04 (s, 1H, ArH), 7.05–6.96 (m, 5H, ArH), 6.79–6.74 (m, 2H,
ArH), 5.86 (s, 1H, NHAr), 4.14 (s, 2H, CH ), 2.69 (d, J = 16.5 Hz, 3H,
CH ); Z-isomer: 11.72 (s, 1H, NHN), 7.61 (d, J = 9.0 Hz, 2H, ArH),
.32 (t, J = 7.9 Hz, 2H, ArH), 7.09 (t, J = 7.4 Hz, 1H, ArH), 7.04 (s, 1H,
ArH), 7.05–6.96 (m, 5H, ArH), 6.79–6.74 (m, 2H, ArH), 5.86 (s, 1H,
3
) δ: E-isomer: 11.72 (s, 1H, NHN), 7.61 (d, J =
–1
176.4 ℃; IR (KBr, cm ) v: 3292, 1629, 1604, 1503, 1490, 1420,
9
1
3
1239, 1169, 917, 868; H NMR(400 MHz, CDCl ) δ: E-isomer; 11.48
(
s, 1H, NHN), 7.60 (dd, J = 9.0, 2.5 Hz, 2H, ArH), 7.32 (t, J = 8.0 Hz,
2
2
7
1
2
2
H, ArH), 7.09 (d, J = 7.4 Hz, 1H, ArH), 7.04 (d, J = 2.1 Hz, 1H, ArH),
.02 (d, J = 2.2 Hz, 1H, ArH), 7.00 (d, J = 0.9 Hz, 1H, ArH), 6.98 (s,
H, ArH), 4.08 (s, 2H, CH
.70 (s, 3H, CCH ); Z-isomer: 11.72 (s, 1H, NHN), 7.60 (dd, J = 9.0,
.5 Hz, 2H, ArH), 7.32 (t, J = 8.0 Hz, 2H, ArH), 7.09 (d, J = 7.4 Hz, 1H,
3
7
2 3
), 3.86 (s, 1H, NHC), 2.76 (s, 3H, NCH ),
3
1
3
NHAr), 4.14 (s, 2H, CH
101MHz, CDCl ) δ: E-isomer: 190.66, 171.78, 169.99, 157.83 (d, J
9.0 Hz), 157.79, 152.18, 144.06, 135.50, 130.45, 123.44, 121.07,
2 3
), 2.69 (d, J = 16.5 Hz, 3H, CH ); C NMR
(
3
ArH), 7.04 (d, J = 2.1 Hz, 1H, ArH), 7.02 (d, J = 2.2 Hz, 1H, ArH), 7.00
=
1
9
1
1
(
d, J = 0.9 Hz, 1H, ArH), 6.98 (s, 1H, ArH), 4.66 (s, 1H, NHC), 4.08 (s,
19.92, 118.38, 116.36 (d, J = 22.7 Hz), 114.74 (d, J = 7.7 Hz),
5.64, 55.85, 13.49; Z-isomer: 193.61, 170.95, 168.73, 157.88,
57.83 (d, J = 9.0 Hz). 151.90, 144.06, 136.22, 130.45, 123.51,
20.94, 119.92, 118.32, 116.36 (d, J = 22.7 Hz), 114.78 (d, J = 6.8
13
2 3 3
2H, CH ), 2.76 (s, 3H, NCH ), 2.70 (s, 3H, CCH ); C NMR (101MHz,
CDCl
3
) δ: E-isomer: 193.06, 169.32, 157.67, 151.05, 135.54,
1
3
1
3
3
29.73, 123.00, 121.31, 120.77, 119.78, 118.30, 55.83, 44.94,
9.19, 12.35; Z-isomer: 193.06, 169.32, 157.67, 152.86, 135.54,
29.79, 123.20, 121.31, 120.77, 119.65, 118.57, 55.83, 44.94,
7.08, 13.55; HR-MS (ESI): m/z calcd for C19H N O ([M + H] )
20 3 3
38.1499, found 338.1500.
Hz), 97.01, 53.82, 12.47; HR-MS (ESI): m/z calcd for C24
H21FN
3
O
3
+
(
[M + H] ) 418.1561, found 418.1564.
+
3
-(1-(2-(2-Chlorophenyl)hydrazinyl)ethylidene)-1-(4-phenoxy-
phenyl)pyrrolidine-2,4-dione (4f). Gray solid, yield 35.5%, m.p.
72.6–174.1 ℃; IR (KBr, cm ) v: 3270, 3113, 1682, 1631, 1595,
564, 1488, 1448, 1375, 1222, 1166, 938, 746; H NMR(400 MHz,
3
) δ: E-isomer: 11.65 (s, 1H, NHN), 7.62 (d, J = 8.6 Hz, 2H, ArH),
.35–7.30 (m, 3H, ArH), 7.22 (t, J = 7.6 Hz, 1H, ArH), 7.09 (t, J = 7.4
Hz, 1H, ArH), 7.04 (d, J = 9.0 Hz, 2H, ArH), 7.00 (d, J = 7.9 Hz, 2H,
ArH), 6.93 (t, J = 7.4 Hz, 1H, ArH), 6.83 (d, J = 8.0 Hz, 1H), 6.37 (s,
2 3
H, NHAr), 4.15 (s, 2H, CH ), 2.70 (s, 3H, CH ); Z-isomer: 11.91 (s,
1
-(4-Phenoxyphenyl)-3-(1-(2-phenylhydrazinyl)ethylidene)-
pyrrolidine-2,4-dione (4c). Light yellow solid, yield 60.4%, m.p.
–1
1
1
CDCl
7
1
–1
172.5–173.6 ℃; IR (KBr, cm ) v: 3256, 3129, 1680, 1626, 1603,
1
1
578, 1488, 1451, 1374, 1357, 1226, 1165, 867, 826, 780; H
) δ: E-isomer: 11.72 (s, 1H, NHN), 7.62 (d, J =
.0 Hz, 2H, ArH), 7.35–7.27 (m, 4H, ArH), 7.11–6.98 (m, 6H, ArH),
.80 (d, J = 7.7 Hz, 2H, ArH), 5.91 (s, 1H, NHAr), 4.14 (s, 2H, CH ),
NMR(400 MHz, CDCl
9
6
3
2
1
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 7
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Ple Na es we dJ oo u nr no at l ao df j Cu hs te mm ias tr rg yins
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Journal Name
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1
2
3
4
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6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
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5
5
5
5
6
1
7
9
H, NHN), 7.62 (d, J = 8.6 Hz, 2H, ArH), 7.35–7.30 (m, 3H, ArH),
.22 (t, J = 7.6 Hz, 1H, ArH), 7.09 (t, J = 7.4 Hz, 1H, ArH), 7.04 (d, J =
.0 Hz, 2H, ArH), 7.00 (d, J = 7.9 Hz, 2H, ArH), 6.93 (t, J = 7.4 Hz, 1H,
3-(1-(2-(2,4-Dichlorophenyl)hydrazinyl)ethylidene)-1-(4-
View Article Online
DOI: 10.1039/D0NJ04551A
phenoxyphenyl)pyrrolidine-2,4-dione (4i). White solid, yield
–
1
63.7%, m.p. 171.2–172.9 ℃; IR (KBr, cm ) v: 3323, 3073, 3040,
1679, 1639, 1590, 1562, 1505, 1489, 1445, 1377, 1231, 1160,
ArH), 6.83 (d, J = 8.0 Hz, 1H), 6.42 (s, 1H, NHAr), 4.15 (s, 2H, CH
2
),
13
1
2
1
1
9
1
1
.74(s, 3H, CH
3
); C NMR (101MHz, CDCl
3
): E-isomer: 191.01,
1046, 938, 837; H NMR(400 MHz, CDCl
3
) δ: E-isomer: 11.65 (s, 1H,
71.82, 171.29, 157.51, 153.22, 142.02, 134.29, 129.78, 128.22,
23.15, 122.36, 121.01, 120.77, 119.72, 119.20, 118.48, 113.28,
6.78, 56.04, 13.42; Z-isomer: 194.15, 172.54, 168.52, 157.64,
52.96, 142.02, 134.95, 129.75, 128.22, 123.04, 122.45, 121.01,
NHN), 7.61 (d, J = 8.2 Hz, 2H, ArH), 7.37 (s, 1H, ArH), 7.33 (t, J = 7.7
Hz, 2H, ArH), 7.19 (d, J = 8.6 Hz, 1H, ArH), 7.09 (t, J = 7.4 Hz, 1H,
ArH), 7.02 (dd, J = 16.1, 8.4 Hz, 4H, ArH), 6.76 (d, J = 8.1 Hz, 1H,
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
2 3
ArH), 6.35 (s, 1H, NHAr), 4.15 (s, 2H, CH ), 2.68 (s, 3H, CH );
20.77, 119.72, 119.20, 118.40, 113.21, 98.26, 54.00, 12.35;
Z-isomer: 11.88 (s, 1H, NHN), 7.61 (d, J = 8.2 Hz, 2H, ArH), 7.37 (s,
1H, ArH), 7.33 (t, J = 7.7 Hz, 2H, ArH), 7.19 (d, J = 8.6 Hz, 1H, ArH),
7.09 (t, J = 7.4 Hz, 1H, ArH), 7.02 (dd, J = 16.1, 8.4 Hz, 4H, ArH),
+
HR-MS (ESI): m/z calcd for C24
34.1268.
-(1-(2-(3-Chlorophenyl)hydrazinyl)ethylidene)-1-(4-phenoxy-
phenyl)pyrrolidine-2,4-dione (4g). Gray solid, solid 45.8%, m.p.
H21ClN
3
O
3
([M + H] ) 434.1266, found
4
3
6.76 (d, J = 8.1 Hz, 1H, ArH), 6.35 (s, 1H, NHAr), 4.15 (s, 2H, CH
2
),
1
3
2.68 (s, 3H, CH ); C NMR (101MHz, CDCl ) δ: E-isomer: 190.26,
3
3
–1
1
1
65.3–166.7 ℃; IR (KBr, cm ) v: 3298, 3271, 3036, 1636, 1597,
171.69, 171.11, 157.46, 153.38, 140.96, 134.11, 129.79, 129.49,
128.26, 126.72, 123.19, 121.02, 119.67, 118.51, 113.96, 96.99,
92.19, 56.10, 13.41; Z-isomer: 190.96, 172.28, 169.05, 157.46,
153.38, 140.96, 134.11, 129.79, 129.49, 128.26, 126.72, 123.19,
503, 1488, 1442, 1374, 1296, 1226, 871; 1H NMR(400 MHz,
CDCl
.36–7.31 (m, 2H, ArH), 7.20 (t, J = 8.0 Hz, 1H, ArH), 7.10 (t, J = 7.4
Hz, 1H, ArH), 7.05 (d, J = 2.1 Hz, 1H, ArH), 7.04–7.03 (m, 1H, ArH),
3
) δ: E-isomer: 11.64 (s, 1H, NHN), 7.62 (d, J = 8.7 Hz, 2H, ArH),
7
120.88, 119.62, 118.51, 113.96, 96.99, 92.19, 56.05, 13.34; HR-MS
+
7
1
.01 (d, J = 1.0 Hz, 1H, ArH), 6.99 (s, 1H, ArH), 6.96 (d, J = 7.5 Hz,
H, ArH), 6.78 (s, 1H, ArH), 6.67 (dd, J = 8.1, 1.6 Hz, 1H, ArH), 6.00
(ESI): m/z calcd for C24
H
20Cl
2
N
3
O
3
([M + H] ) 468.0876, found
468.0873.
(
(
s, 1H, NHAr), 4.14 (s, 2H, CH
s, 1H, NHN), 7.62 (d, J = 8.7 Hz, 2H, ArH), 7.36–7.31 (m, 2H, ArH),
2
), 2.67 (s, 3H, CH
3
); Z-isomer: 11.90
3-(1-(2-(4-Bromophenyl)hydrazinyl)ethylidene)-1-(4-phenoxy-
phenyl)pyrrolidine-2,4-dione (4j). White solid, yield 50.2%, m.p.
156.3–158.2 ℃; IR (KBr, cm ) v: 3248, 3101, 1642, 1602, 1507,
–1
7
2
.20 (t, J = 8.0 Hz, 1H, ArH), 7.10 (t, J = 7.4 Hz, 1H, ArH), 7.05 (d, J =
.1 Hz, 1H, ArH), 7.04–7.03 (m, 1H, ArH), 7.01 (d, J = 1.0 Hz, 1H,
1
1489, 1433, 1382, 1241, 1174, 872, 818; H NMR(400 MHz, CDCl
3
)
ArH), 6.99 (s, 1H, ArH), 6.96 (d, J = 7.5 Hz, 1H, ArH), 6.78 (s, 1H,
ArH), 6.67 (dd, J = 8.1, 1.6 Hz, 1H, ArH), 6.00 (s, 1H, NHAr), 4.14 (s,
δ: E-isomer: 11.67 (s, 1H, NHN), 7.61 (d, J = 8.9 Hz, 2H, ArH), 7.39
(d, J = 8.7 Hz, 2H, ArH), 7.33 (dd, J = 8.5, 7.5 Hz, 2H, ArH), 7.09 (t, J
= 7.4 Hz, 1H, ArH), 7.05 (s, 1H, ArH), 7.02 (s, 1H, ArH), 7.01 (d, J =
1.0 Hz, 1H, ArH), 6.98 (s, 1H, ArH), 6.69 (d, J = 8.8 Hz, 2H, ArH),
1
3
2
1
1
1
1
1
1
4
2 3 3
H, CH ), 2.72 (s, 3H, CH ); C NMR (101MHz, CDCl ) δ: E-isomer:
91.11, 171.84, 171.20, 157.39, 153.44, 147.42, 135.45, 130.68,
29.81, 123.25, 122.01, 121.33, 121.05, 119.68, 118.56, 113.29,
11.29, 96.57, 56.11, 13.44; Z-isomer: 194.12, 172.33, 168.71,
57.50, 153.20, 147.27, 135.50, 130.71, 129.78, 123.14, 122.11,
21.33, 121.05, 119.66, 118.49, 113.29, 111.29, 98.07, 54.12,
2.37; HR-MS (ESI): m/z calcd for C24
2 3
5.92 (s, 1H, NHAr), 4.14 (s, 2H, CH ), 2.68 (s, 3H, CH ); Z-isomer:
11.67 (s, 1H, NHN), 7.61 (d, J = 8.9 Hz, 2H, ArH), 7.39 (d, J = 8.7 Hz,
2H, ArH), 7.33 (dd, J = 8.5, 7.5 Hz, 2H, ArH), 7.09 (t, J = 7.4 Hz, 1H,
ArH), 7.05 (s, 1H, ArH), 7.02 (s, 1H, ArH), 7.01 (d, J = 1.0 Hz, 1H,
ArH), 6.98 (s, 1H, ArH), 6.69 (d, J = 8.8 Hz, 2H, ArH), 5.92 (s, 1H,
), 2.68 (s, 3H, CH ); C NMR (101MHz,
2 3
CDCl ) δ: E-isomer: 190.88, 171.86, 171.37, 156.23, 152.76,
142.04, 134.67, 129.79, 129.73, 128.22, 122.41, 120.78, 119.79,
119.61, 113.28, 96.76, 55.99, 13.44; Z-isomer: 194.06, 172.60,
168.54, 156.36, 152.52, 141.97, 135.33, 129.82, 129.70, 128.08,
+
H21ClN
3
O
3
([M + H] )
1
3
34.1266, found 434.1263.
NHAr), 4.14 (s, 2H, CH
3-(1-(2-(4-Chlorophenyl)hydrazinyl)ethylidene)-1-(4-phenoxy-
3
phenyl)pyrrolidine-2,4-dione (4h). White solid, yield 62.1%, m.p.
–1
178.6–180.3 ℃; IR (KBr, cm ) v: 3482, 3413, 3234, 1632, 1617,
1
1599, 1504, 1487, 1232, 945, 814; H NMR(400 MHz, CDCl
3
) δ:
E-isomer: 11.65 (s, 1H, NHN), 7.61 (d, J = 8.8 Hz, 2H, ArH), 7.33 (dd,
J = 8.4, 7.5 Hz, 2H, ArH), 7.26 (s, 1H, ArH), 7.23 (s, 1H, ArH), 7.09 (t,
J = 7.4 Hz, 1H, ArH), 7.05 (s, 1H, ArH), 7.02 (s, 1H, ArH), 7.00 (d, J =
122.50, 121.02, 119.54, 119.22, 113.22, 98.24, 53.95, 12.37;
+
HR-MS (ESI): m/z calcd for C24
found 478.0759.
H
21BrN
3
O
3
([M + H] ) 478.0761,
1
5
.0 Hz, 1H, ArH), 6.98 (s, 1H, ArH), 6.73 (d, J = 8.8 Hz, 2H, ArH),
.93 (s, 1H, NHAr), 4.14 (s, 2H, CH ), 2.68 (s, 3H, CH );
1-(4-Phenoxyphenyl)-3-(1-(2-(4-(trifluoromethyl)phenyl)-
hydrazinyl)ethylidene)pyrrolidine-2,4-dione (4k). Yellow solid,
yield 36.9%, m.p. 143.5–143.9 ℃; IR (KBr, cm ) v: 3311, 3246,
3059, 1642, 1619, 1576, 1507, 1489,1431, 1383, 1326, 1238, 1104,
2
3
–
1
Z-isomer:11.91 (s, 1H, NHN), 7.61 (d, J = 8.8 Hz, 2H, ArH), 7.33 (dd,
J = 8.4, 7.5 Hz, 2H, ArH), 7.26 (s, 1H, ArH), 7.23 (s, 1H, ArH), 7.09 (t,
J = 7.4 Hz, 1H, ArH), 7.05 (s, 1H, ArH), 7.02 (s, 1H, ArH), 7.00 (d, J =
1
1066, 828, 743; H NMR(400 MHz, CDCl
3
) δ: E-isomer: 11.69 (s, 1H,
1.0 Hz, 1H, ArH), 6.98 (s, 1H, ArH), 6.73 (d, J = 8.8 Hz, 2H, ArH),
NHN), 7.61 (d, J = 9.0 Hz, 2H, ArH), 7.54 (d, J = 8.5 Hz, 2H, ArH),
7.36–7.30 (m, 2H, ArH), 7.09 (t, J = 7.4 Hz, 1H, ArH), 7.05 (d, J = 2.0
Hz, 1H, ArH), 7.03 (d, J = 2.1 Hz, 1H, ArH), 7.01 (d, J = 1.0 Hz, 1H,
ArH), 6.99 (d, J = 0.8 Hz, 1H, ArH), 6.85 (d, J = 8.5 Hz, 2H, ArH), 6.19
13
5.93 (s, 1H, NHAr), 4.14 (s, 2H, CH
2 3
), 2.68 (s, 3H, CH ); C NMR
) δ: E-isomer: 191.06, 171.89, 171.09, 157.41,
(
101MHz, CDCl
3
1
1
1
1
1
4
53.41, 144.68, 134.13, 129.81, 127.02, 123.25, 121.26, 121.03,
19.68, 118.54, 114.49, 96.48, 56.10, 13.44; Z-isomer: 194.07,
72.22, 168.72, 157.50, 153.17, 144.55, 134.73, 129.57, 127.11,
23.15, 121.26, 121.03, 119.68, 118.48, 114.49, 97.98, 54.09,
2.38; HR-MS (ESI): m/z calcd for C24
2 3
(s, 1H, NHAr), 4.15 (s, 2H, CH ), 2.68 (s, 3H, CH ); Z-isomer: 11.92
(s, 1H, NHN), 7.61 (d, J = 9.0 Hz, 2H, ArH), 7.54 (d, J = 8.5 Hz, 2H,
ArH), 7.36–7.30 (m, 2H, ArH), 7.09 (t, J = 7.4 Hz, 1H, ArH), 7.05 (d, J
= 2.0 Hz, 1H, ArH), 7.03 (d, J = 2.1 Hz, 1H, ArH), 7.01 (d, J = 1.0 Hz,
1H, ArH), 6.99 (d, J = 0.8 Hz, 1H, ArH), 6.85 (d, J = 8.5 Hz, 2H, ArH),
+
H21ClN
3
O
3
([M + H] )
34.1266, found 434.1263.
13
6
2 3
.19 (s, 1H, NHAr), 4.15 (s, 2H, CH ), 2.68 (s, 3H, CH ); C NMR
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 8
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Page 9 of 13
Ple Na es we dJ oo u nr no at l ao df j Cu hs te mm ias tr rg yins
Journal Name
ARTICLE
1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
(
101MHz, CDCl
3
) δ: E-isomer: 191.13, 171.78, 171.17, 157.39,
(d, J = 4.3 Hz), 134.67, 129.72, 128.08, 125.00, 122.25, 121.04,
View Article Online
DOI: 10.1039/D0NJ04551A
1
1
9
1
1
9
53.58, 148.91, 133.98, 129.82, 127.01 (d, J = 3.6 Hz), 125.57,
120.81, 119.77, 119.59, 115.50( d, J = 19.1 Hz), 114.23, 96.57,
23.92 (t, J = 135.6 Hz), 121.42, 121.15, 119.65, 118.57, 112.61,
6.78, 56.17, 13.39; E-isomer:192.64, 172.29, 168.62, 157.35,
53.38, 148.73, 134.54, 129.82, 127.01 (d, J = 3.6 Hz), 125.57,
23.92 (t, J = 135.6 Hz), 121.42, 121.15, 119.65, 118.57, 112.61,
55.95, 13.44; Z-isomer: 193.99, 172.63, 168.64, 156.34, 152.51,
152.23 (d, J = 4.3 Hz), 134.67, 129.72, 127.95, 125.00, 122.16,
121.04, 120.81, 119.77, 119.59, 115.50 (d, J = 19.1 Hz),
114.18,96.35, 53.92, 12.37; HR-MS (ESI): m/z calcd for
+
8.23, 54.16, 12.37; HR-MS (ESI): m/z calcd for C25
+
H
21
3
F N
O
3 3
([M +
C
24
H
20ClFN
3
O
3
([M + H] ) 425.1171, found 452.1164.
H] ) 468.1530, found 468.1530.
1-(4-(4-Chlorophenoxy)phenyl)-3-(1-(2-(4-fluorophenyl)-
hydrazinyl)ethylidene)pyrrolidine-2,4-dione (4o). Yellow, yield
50.3%, m.p. 194.2–195.7 ℃; IR (KBr, cm ) v: 3308, 3255, 3084,
1685, 1630, 1578, 1503, 1484, 1419, 1378, 1238, 821; H NMR(400
3
MHz, CDCl ) δ: E-isomer: 11.67 (s, 1H, NHN), 7.62 (d, J = 8.9 Hz, 2H,
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1-(4-(4-Chlorophenoxy)phenyl)-3-(1-(2-methylhydrazinyl)-
–1
ethylidene)pyrrolidine-2,4-dione (4l). Green solid, yield 65.2%,
m.p. 160.6–161.8 ℃; IR (KBr, cm ) v: 3288, 3059, 1671, 1632,
–1
1
1
601, 1501, 1484, 1447, 1388, 1377, 1238, 1164, 1078, 919, 831;
1
H NMR(400 MHz, CDCl
H, ArH), 7.28 (s, 1H, ArH), 7.26 (s, 1H, ArH), 7.01 (d, J = 8.8 Hz, 2H,
ArH), 6.91 (d, J = 8.8 Hz, 2H, ArH), 4.07 (s, 2H, CH ), 3.86 (s, 1H,
NHAr), 2.76 (s, 3H, NCH ), 2.70 (s, 3H, CH ); Z-isomer: 11.80 (s, 1H,
3
) δ: E-isomer: 11.42 (s, 1H, NHN), 7.61 (s,
ArH), 7.28 (d, J = 2.1 Hz, 2H, ArH), 7.04–6.97 (m, 4H, ArH), 6.93 (d,
J = 2.1 Hz, 1H, ArH), 6.91 (d, J = 2.1 Hz, 1H, ArH), 6.79–6.74 (m, 2H,
2
2
ArH), 5.85 (s, 1H, NHAr), 4.12 (d, J = 9.2 Hz, 2H, CH
2
), 2.71 (s, 3H,
3
3
CH ); Z-isomer: 11.93 (s, 1H, NHN), 7.62 (d, J = 8.9 Hz, 2H, ArH),
7.28 (d, J = 2.1 Hz, 2H, ArH), 7.04–6.97 (m, 4H, ArH), 6.93 (d, J = 2.1
Hz, 1H, ArH), 6.91 (d, J = 2.1 Hz, 1H, ArH), 6.79–6.74 (m, 2H, ArH),
3
NHN), 7.61 (s, 2H, ArH), 7.28 (s, 1H, ArH), 7.26 (s, 1H, ArH), 7.01 (d,
J = 8.8 Hz, 2H, ArH), 6.91 (d, J = 8.8 Hz, 2H, ArH), 4.66 (s, 1H,
1
3
NHAr), 4.07 (s, 2H, CH
NMR (101MHz, CDCl ) δ: E-isomer: 190.60, 172.50, 169.37, 156.35,
52.42, 134.99, 129.68, 127.94, 120.76, 119.82, 119.47, 94.96,
2
), 2.76 (s, 3H, NCH
3
), 2.70 (s, 3H, CH
3
);
C
5.85 (s, 1H, NHAr), 4.12 (d, J = 9.2 Hz, 2H, CH
2
), 2.71 (s, 3H, CH
3
);
1
3
3
C NMR (101MHz, CDCl ) δ: E-isomer:190.96, 171.99, 168.33,
3
1
5
1
5
159.60, 157.21, 156.21, 152.76, 142.07, 129.73, 128.09, 120.99,
119.75, 119.61, 116.32 (d, J = 23.0 Hz),114.74 (d, J = 7.9 Hz), 96.31,
56.00, 13.43.; Z-isomer: 194.19, 171.99, 168.33, 159.60, 157.21,
156.21, 152.76, 142.07, 129.73, 128.09, 120.99, 119.75, 119.61,
5.73, 39.15, 13.53; Z-isomer: 193.06, 172.50, 170.36, 156.50,
52.08, 135.69, 129.64, 127.79, 120.76, 119.82, 119.37, 96.60,
3.76, 39.21, 12.39; HR-MS (ESI): m/z calcd for C19
+
H19ClN
3
O
3
([M +
H] ) 372.1109, found 372.1473.
116.32 (d, J = 23.0 Hz), 114.74 (d, J = 7.9 Hz), 96.31, 56.00, 13.43;
+
1-(4-(4-Chlorophenoxy)phenyl)-3-(1-(2-phenylhydrazinyl)-
HR-MS (ESI): m/z calcd for C24
H
20ClFN
3
O
3
([M + H] ) 425.1171,
ethylidene)pyrrolidine-2,4-dione (4m). Gray solid, yield 35.2%,
m.p. 174.4–176.4 ℃; IR (KBr, cm ) v: 3254, 1629, 1606, 1577,
found 452.1164.
–1
1-(4-(4-Chlorophenoxy)phenyl)-3-(1-(2-(4-chlorophenyl)-
hydrazinyl)ethylidene)pyrrolidine-2,4-dione (4p). Gray solid, yield
1
1502, 1486, 1410, 1378, 1239, 1169, 829; H NMR(400 MHz, CDCl
3
)
–1
δ: E-isomer: 11.67 (s, 1H, NHN), 7.64 (d, J = 8.7 Hz, 2H, ArH), 7.30
dd, J = 10.5, 5.1 Hz, 4H, ArH), 7.02 (dd, J = 11.9, 5.0 Hz, 3H, ArH),
58.2%, m.p. 159.1–159.8 ℃; IR (KBr, cm ) v: 3250, 1683, 1628,
1
(
1572, 1501, 1484, 1445, 1375, 1240, 1088, 946, 848, 819; H
6
=
CH
7
.94 (d, J = 2.0 Hz, 1H, ArH), 6.92 (d, J = 2.1 Hz, 1H, ArH), 6.81 (d, J
7.8 Hz, 2H, ArH), 5.89 (s, 1H, NHAr), 4.14 (s, 2H, CH ), 2.72 (s, 3H,
); Z-isomer: 11.95 (s, 1H, NHN), 7.64 (d, J = 8.7 Hz, 2H, ArH),
.30 (dd, J = 10.5, 5.1 Hz, 4H, ArH), 7.02 (dd, J = 11.9, 5.0 Hz, 3H,
ArH), 6.94 (d, J = 2.0 Hz, 1H, ArH), 6.92 (d, J = 2.1 Hz, 1H, ArH), 6.81
3
NMR(400 MHz, CDCl ) δ: E-isomer: 11.66 (s, 1H, NHN), 7.62 (d, J =
8.4 Hz, 2H, ArH), 7.29 (s, 2H, ArH), 7.23 (s, 2H, ArH), 7.02 (d, J = 8.8
Hz, 2H, ArH), 6.92 (d, J = 8.8 Hz, 2H, ArH), 6.74 (d, J = 8.4 Hz, 2H,
2
3
2 3
ArH), 5.92 (s, 1H, NHAr), 4.14 (s, 2H, CH ), 2.69 (s, 3H, CH );
Z-isomer: 11.91 (s, 1H, NHN), 7.62 (d, J = 8.4 Hz, 2H, ArH), 7.29 (s,
2H, ArH), 7.23 (s, 2H, ArH), 7.02 (d, J = 8.8 Hz, 2H, ArH), 6.92 (d, J =
8.8 Hz, 2H, ArH), 6.74 (d, J = 8.4 Hz, 2H, ArH), 5.92 (s, 1H, NHAr),
(
(
1
1
d, J = 7.8 Hz, 2H, ArH), 5.89 (s, 1H, NHAr), 4.14 (s, 2H, CH
s, 3H, CH ); C NMR (101MHz, CDCl
3 3
2
), 2.72
) δ: E-isomer: 190.94, 172.03,
71.31, 156.24, 152.74, 145.96, 134.71, 129.73, 128.08, 122.20,
21.13, 120.87, 119.78, 119.60, 113.29, 96.26, 55.97, 13.51;
1
3
1
3
2 3 3
4.14 (s, 2H, CH ), 2.69 (s, 3H, CH ); C NMR (101MHz, CDCl ) δ:
E-isomer: 190.97, 171.91, 171.16, 156.13, 152.92, 144.65, 134.51,
129.76, 129.74, 128.16, 127.05, 121.00, 119.75, 119.67, 114.51,
96.46, 56.04, 13.48; Z-isomer: 191.44, 172.26, 168.76, 156.22,
152.73, 144.53, 135.10, 129.74, 129.59, 128.07, 127.14, 121.28,
119.71, 119.63, 114.51, 97.93, 56.04, 12.45; HR-MS (ESI): m/z
([M + H] ) 468.0876, found 468.0871.
1-(4-(4-Chlorophenoxy)phenyl)-3-(1-(2-(2,4-dichlorophenyl)-
hydrazinyl)ethylidene)pyrrolidine-2,4-dione (4q). Gray solid, yield
60.7%, m.p. 185.3–186.0 ℃; IR (KBr, cm ) v: 3259, 3150, 1689,
1634, 1591, 1560, 1501, 1484, 1444, 1373, 1232, 1163, 1051, 950,
Z-isomer: 193.82, 172.44, 168.89, 156.31, 152.53, 145.88, 135.33,
129.66, 127.99, 122.27, 121.13, 120.87, 119.78, 119.60, 113.29,
9
7.77, 53.98, 12.43; HR-MS (ESI): m/z calcd for C24
H21ClN
3
O
3
([M +
+
H] ) 434.1266, found 434.1259.
+
1-(4-(4-Chlorophenoxy)phenyl)-3-(1-(2-(2-fluorophenyl)-
calcd for C24
H
20Cl
N
2 3
O
3
hydrazinyl)ethylidene)pyrrolidine-2,4-dione (4n). Gray solid, yield
–1
48.9%, m.p. 196.4.4–198.2 ℃; IR (KBr, cm ) v: 3298, 3106, 1670
1
–1
1636, 1578, 1484, 1447, 1384, 1240, 1192, 826; H NMR(400 MHz,
CDCl
3
) δ: E-isomer: 11.62 (s, 1H, NHN), 7.64 (s, 2H, ArH), 7.28 (s,
1
2H, ArH), 7.09 (t, J = 9.9 Hz, 2H, ArH), 7.02 (d, J = 8.9 Hz, 2H, ArH),
847, 810; H NMR(400 MHz, CDCl
3
) δ: E-isomer: 11.63 (s, 1H,
6.92 (d, J = 8.8 Hz, 3H, ArH), 6.86 (d, J = 7.4 Hz, 1H, ArH), 6.15 (d, J
NHN), 7.63 (t, J = 8.6 Hz, 2H, ArH), 7.37 (s, 1H, ArH), 7.29 (s, 2H,
ArH), 7.19 (d, J = 8.6 Hz, 1H, ArH), 7.03 (d, J = 9.0 Hz, 2H, ArH), 6.92
(d, J = 8.8 Hz, 2H, ArH), 6.77 (d, J = 8.8 Hz, 1H, ArH), 6.35 (d, J =
=
2 3
15.2 Hz, 1H, NHAr), 4.14 (s, 2H, CH ), 2.73 (s, 3H, CH ); Z-isomer:
1
9
3
1.90 (s, 1H, NHN), 7.62 (s, 2H, ArH), 7.28 (s, 2H, ArH), 7.09 (t, J =
.9 Hz, 2H, ArH), 7.02 (d, J = 8.9 Hz, 2H, ArH), 6.92 (d, J = 8.8 Hz,
H, ArH), 6.86 (d, J = 7.4 Hz, 1H, ArH), 6.15 (d, J = 15.2 Hz, 1H,
2 3
19.4 Hz, 1H, NHAr), 4.14 (s, 2H, CH ), 2.68 (s, 3H, CH ); Z-isomer:
11.89 (s, 1H, NHN), 7.63 (t, J = 8.6 Hz, 2H, ArH), 7.37 (s, 1H, ArH),
7.29 (s, 2H, ArH), 7.19 (d, J = 8.6 Hz, 1H, ArH), 7.03 (d, J = 9.0 Hz,
2H, ArH), 6.92 (d, J = 8.8 Hz, 2H, ArH), 6.77 (d, J = 8.8 Hz, 1H, ArH),
1
3
2 3
NHAr), 4.14 (s, 2H, CH ), 2.76 (s, 3H, CH ); C NMR (101MHz,
CDCl ) δ: E-isomer: 190.91, 171.90, 171.43, 156.22, 152.75, 152.23
3
This journal is © The Royal Society of Chemistry 20xx
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1
2
3
4
5
6
7
8
9
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
5
5
6
6
.35 (d, J = 19.4 Hz, 1H, NHAr), 4.16 (s, 2H, CH
C NMR (101MHz, CDCl
2
), 2.70 (s, 3H, CH
) δ: E-isomer: 190.89, 171.74, 171.15,
3
);
mycelim growth rate method.39,40 Under the same conditions, the
View Article Online
1
3
DOI: 10.1039/D0NJ04551A
3
agricultural fungicides carbendazim and boscalid were used as the
1
1
56.16, 152.91, 140.85, 134.51, 129.74, 129.50, 128.28, 128.02,
26.79, 120.87, 119.76, 119.66, 114.00, 98.48, 97.00, 56.04, 13.39;
positive controls to assess the antifungal effect of target
compounds. The anti-Rs EC50 values of target compounds were
calculated under the basic of the corresponding antifungal effects
at five double-declining concentrations. The in vivo anti-Rs effect
of target compound 4h was evaluated on rice leaves by a
detached leaf assay, with the commercial fungicides boscalid,
Z-isomer: 194.19, 172.40, 168.34, 156.29, 152.64, 140.81, 135.18,
129.71, 129.53, 128.15, 128.02, 126.90, 121.09, 119.66, 119.57,
113.95, 98.48, 97.00, 53.94, 12.34; HR-MS (ESI): m/z calcd for
+
C
24
3
H19Cl N
3
O
3
([M + H] ) 502.0486, found 502.0482.
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
4
1
3-(1-(2-(4-Bromophenyl)hydrazinyl)ethylidene)-1-(4-(4-chloro-
carbendazim, and hymexazol as the positive controls. To
preliminarily investigate the antifungal mode of compound 4h
against Rs, the hypha morphology was observed by a scanning
phenoxy)phenyl)pyrrolidine-2,4-dione (4r). Gray solid, yield
–
1
25.3%, m.p. 166.8–167.8 ℃; IR (KBr, cm ) v: 3255, 3166, 1686,
1
42
1629, 1598, 1568, 1485, 1444, 1372, 1234, 1005, 954, 848, 810; H
electron microscope.
NMR(400 MHz, CDCl
3
) δ: E-isomer: 11.63 (s, 1H, NHN), 7.62 (d, J =
8
2
6
1
1
7
=
2
.6 Hz, 2H, ArH), 7.39 (d, J = 8.6 Hz, 2H, ArH), 7.28 (d, J = 2.1 Hz,
H, ArH), 7.02 (d, J = 9.0 Hz, 2H, ArH), 6.93 (d, J = 2.1 Hz, 1H, ArH),
.91 (d, J = 2.1 Hz, 1H, ArH), 6.69 (d, J = 8.7 Hz, 2H, ArH), 5.92 (s,
Study of 3D-QSAR model
The anti-Rs EC50 values (mmol/mL) of target compounds 4a–4s
were calculated as the negative logarithm (pEC50) for building a
three-dimensional structrue-activity relationship (3D-QSAR) model
under the SYBYL 2.0 software (Shanghai. Tri-I. Biotech. Inc.,
2 3
H, NHAr), 4.14 (s, 2H, CH ), 2.68 (s, 3H, CH ); Z-isomer: 11.91 (s,
H, NHN), 7.62 (d, J = 8.6 Hz, 2H, ArH), 7.39 (d, J = 8.6 Hz, 2H, ArH),
.28 (d, J = 2.1 Hz, 2H, ArH), 7.02 (d, J = 9.0 Hz, 2H, ArH), 6.93 (d, J
2.1 Hz, 1H, ArH), 6.91 (d, J = 2.1 Hz, 1H, ArH), 6.69 (d, J = 8.7 Hz,
H, ArH), 5.92 (s, 1H, NHAr), 4.14 (s, 2H, CH
) δ: E-isomer: 190.96, 171.90, 171.15, 156.12,
52.94, 145.15, 134.49, 132.49, 129.76, 128.17, 121.28, 121.01,
4
3,44
China).
The
molecular
energy
optimization
of
1
3
2 3
), 2.68 (s, 3H, CH ); C
three-dimensional conformations with a convergence criterion of
.005 kcal/mol Å convergence gradient was obtained by using
the Gasteiger-Huckel charges, Tripos force field and Powell
conjugate gradient algorithm. The field properties in CoMSIA
model were calculated by using a partial least squares analysis
PLS). Afterwards, PLS results and the standard deviations of
CoMSIA description values were used to generate the
corresponding CoMSIA contour maps.
NMR (101MHz, CDCl
3
–1
0
1
1
1
1
1
5
19.67, 114.87, 114.30, 96.49, 56.04, 13.46; Z-isomer: 194.02,
71.90, 171.15, 156.20, 152.72, 145.02, 135.08, 132.49, 129.76,
28.08, 121.28, 121.01, 119.74, 114.87, 114.36, 97.98, 54.04,
2.49; HR-MS (ESI): m/z calcd for C24
4
5
4
6
(
+
H
20BrClN
O
3 3
([M + H] )
14.0351, found 514.0346.
1-(4-(4-Chlorophenoxy)phenyl)-3-(1-(2-(4-(trifluoromethyl)-
phenyl)hydrazinyl)ethylidene)pyrrolidine-2,4-dione (4s). Gray
solid, yield 21.5%, m.p. 166.2–167.8 ℃; IR (KBr, cm ) v: 3270,
–1
Conclusions
3073, 2913, 1685, 1637, 1618, 1578, 1504, 1486, 1440, 1420,
In summary, nineteen novel pyrrolidine-2,4-dione derivatives
containing both pharmacophores of hydrazine and diphenyl ether
were designed, synthesized, and characterized. All target
compounds were tested for their antifungal activities against Rs,
Bc, and Fg. The EC50 values of compounds 4d, 4e, 4h, 4j, 4k, 4p,
and 4r against Rs reached respectively 1.31, 1.75, 0.39, 0.50, 2.14,
1
1382, 1329, 1248, 1108, 1068, 945, 829; H NMR(400 MHz, CDCl
3
)
δ: E-isomer: 11.69 (s, 1H, NHN), 7.63 (d, J = 8.3 Hz, 2H, ArH), 7.55
d, J = 8.1 Hz, 2H, ArH), 7.29 (s, 2H, ArH), 7.03 (d, J = 9.0 Hz, 2H,
ArH), 6.92 (d, J = 8.8 Hz, 2H, ArH), 6.86 (d, J = 8.4 Hz, 2H, ArH), 6.12
(
(
(
s, 1H, NHAr), 4.15 (s, 2H, CH
s, 1H, NHN), 7.63 (d, J = 8.3 Hz, 2H, ArH), 7.55 (d, J = 8.1 Hz, 2H,
2 3
), 2.68 (s, 3H, CH ); Z-isomer: 11.95
1
.01, and 0.97 μg/mL, which were obviously superior to that of
ArH), 7.29 (s, 2H, ArH), 7.03 (d, J = 9.0 Hz, 2H, ArH), 6.92 (d, J = 8.8
Hz, 2H, ArH), 6.86 (d, J = 8.4 Hz, 2H, ArH), 6.12 (s, 1H, NHAr), 4.15
boscolid (2.21 μg/mL). The CoMSIA model was established to
further analyze the 3D-QSAR and predict antifungal activity against
Rs. The results showed that introducing a substituted phenyl
1
3
(
s, 2H, CH
2 3 3
), 2.71 (s, 3H, CH ); C NMR (101MHz, CDCl ) δ:
E-isomer: 191.02, 171.80, 171.21, 156.10, 153.04, 148.82, 135.00,
2
fragment, especially 4-FPh, 4-ClPh, or 4-BrPh fragment, at the R
1
1
1
1
1
34.42, 129.76, 128.21, 127.04 (dd, J = 6.7, 3.0 Hz), 121.35,
21.06, 119.75, 119.69, 112.65, 96.82, 56.10, 13.39; Z-isomer:
94.17, 172.35, 168.37 (d, J = 37.3 Hz), 156.19, 152.85, 148.68,
35.00, 134.42, 129.73, 128.11, 127.04 (dd, J = 6.7, 3.0 Hz),
position was necessary to enhance antifungal activity against Rs.
The application effect of compound 4h was proved by the in vivo
bioassay against Rs on rice leaves. Additionally, the SEM
observation indicated that the compound 4h significantly changed
the mycelial morphology. This work indicated that the
pyrrolidine-2,4-dione derivatives containing the hydrazine and
diphenyl ether moieties showed obvious inhibitory activities
against plant pathogenic fungi, which provided a structural
template for developing novel highly-efficiency fungicides.
21.35, 121.06, 119.71, 119.64, 112.65, 96.82, 54.08, 12.34;
+
HR-MS (ESI): m/z calcd for C25
found 502.1136.
H
20ClF
3
N
3
O
3
([M+H] ) 502.1139,
Antifungal activity evaluation
The phytopathogenic fungi strains (Rs, Bc, and Fg) that were
isolated from infected crops in the Middle-lower Yangtze plain
were provided by the Jiangsu Key Laboratory of Pesticide Science
at Nanjing Agriculture University. The inhibition effects of target
compounds 4a–4s against the tested fungi were determined by a
Conflicts of interest
There are no conflicts of interest to declare.
This journal is © The Royal Society of Chemistry 20xx
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Acknowledgements
22 L. Z. Zhang, Z. J. Ren, A. M. Lu, Z. Zhao, W. Q. XU, Q. Q. Bao, W.
View Article Online
DOI: 10.1039/D0NJ04551A
J. Ding and C. L. Yang, Chem. Res. Chin. Univ., 2015, 31(2),
The authors’ work was supported by National Natural Science
Foundation of China (31772209) and the Fundamental Research
Funds for the Central Universities of China (KYTZ201604 and
KJSY201607).
2
28–234.
3 G. H. Lu, H. B. Chu, M. Chen and C, L, Yang, Chin. Chem. Lett.,
014, 25, 61–64.
4 M. Chrysayi-Tokousbalides, M. A. Kastanias, A. Philippoussis
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7 J. A. Sternberg, D. Geffken, J. B. Adams Jr, R. Pöstages, C. G.
2
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DOI: 10.1039/D0NJ04551A
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New Journal of Chemistry
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DOI: 10.1039/D0NJ04551A
Novel
pyrrolidine-2,4-dione
derivatives
containing
both
pharmacophores of hydrazine and diphenyl ether as potential
antifungal agents: design, synthesis, biological evaluation and 3D-QSAR
study
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Hao-Ran Hu, An Wang, Ling-Ling Qiu, Xiao-Bin Wang, Min Chen, Ai-Min Lu and Chun-Long Yang
Novel pyrrolidine-2,4-dione derivatives were designed and synthesized based on natural
compounds and our previous work. Some compounds showed excellent antifungal activity.
Scanning electron microscopy (SEM) was used to observe the changes of mycelium morphology.
3
D-QSAR provided valuable information for further structural optimization.
R1
O
O
O
R2
H
N
N
N
N
N
HN
H
H
O
Br
Diphenyl ether
O
O
Designed molecular skeleton
EC50 = 1.63 ug/mL against Rs
Compd.
R1
H
R2
Anti-Rs EC50 (in vitro)
Improving liposolubility
4h
4-ClPh
0.39 ug/mL
2.21 ug/mL
Our previous work
Boscalid
R
In vivo bioassay
3D-QSA
4
h
hymexazol
boscalid
carbendazim
CK