Design, synthesis, characterization, and biological activities of novel spirooxindole analogues containing hydantoin, thiohydantoin, urea, and thiourea moieties

Article abstract of DOI:10.1021/acs.jafc.0c04488

On the basis of the scaffolds widely used in drug design, a series of novel spirooxindole derivatives containing hydantoin, thiohydantoin, urea, and thiourea moieties have been designed, synthesized, characterized, and first evaluated for their biological activities. The diastereoselectivity mechanism is proposed, and the systematic conformational analysis is performed. The bioassay results show that the target compounds possess moderate to good antiviral activities against tobacco mosaic virus (TMV), among which compound 22 shows the highest antiviral activity in vitro as well as inactivation, curative, and protection activities in vivo (45 ± 1, 47 ± 3, 50 ± 1, and 51 ± 1%, 500 mg/L, respectively), higher than ribavirin (38 ± 1, 36 ± 1, 38 ± 1, and 36 ± 1%, 500 mg/L, respectively). Thus, compound 22 is a promising candidate for anti-TMV development. Most of these compounds show broad-spectrum fungicidal activities against 14 kinds of phytopathogenic fungi and selective fungicidal activities against Physalospora piricola, Sclerotinia sclerotiorum, and Rhizoctonia cerealis. Additionally, some of these compounds exhibit insecticidal activity against Culex pipiens pallens, Mythimna separata, Helicoverpa armigera, and Pyrausta nubilalis. Compound 17 exhibits the highest larvicidal activity (LC₅₀ was 0.32 mg/L) against C. pipiens pallens.

Full text of DOI:10.1021/acs.jafc.0c04488

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Agricultural and Environmental Chemistry
Design, Synthesis, Characterization, and Biological
Activities of Novel Spirooxindoles Analogues Containing
Hydantoin, Thiohydantoin, Urea, and Thiourea Moieties
Hongjian Song, Linwei Chen, Yanke Hao, Yuxiu Liu, Yongqiang Li, Jingjing Zhang, and Qingmin Wang
J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jafc.0c04488 • Publication Date (Web): 31 Aug 2020
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Design, Synthesis, Characterization, and Biological Activities of
Novel Spirooxindoles Analogues Containing Hydantoin,
Thiohydantoin, Urea, and Thiourea Moieties
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Linwei Chen, Yanke Hao, Hongjian Song, Yuxiu Liu, Yongqiang Li, Jingjing
Zhang,1,2* and Qingmin Wang1*
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1State Key Laboratory of Elemento-Organic Chemistry, College of Chemistry,
Collaborative Innovation Center of Chemical Science and Engineering (Tianjin),
Nankai University, Tianjin 300071, People’s Republic of China
2Tianjin Agricultural University, Tianjin 300384, People’s Republic of China
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*To whom correspondence should be addressed. Tel: +86-(0)22-23503952; Fax:
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+86-(0)22-23503952;
E-mail:
songhongjian@nankai.edu.cn;
Email:
chemzhangjingjing@163.com; wangqm@nankai.edu.cn.
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ABSTRACT
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Based on the scaffolds widely used in drug design, a series of novel
spirooxindole derivatives containing hydantoin, thiohydantoin, urea, and thiourea
moieties have been designed, synthesized, characterized, and first evaluated for their
biological activities. The diastereoselectivity mechanism is proposed and the
systematic conformational analysis is performed. The bioassays results show that the
target compounds possess moderate to good antiviral activities against tobacco mosaic
virus (TMV), among which compound 22 shows the highest antiviral activity in vitro
as well as inactivation, curative, and protection activities in vivo (45±1, 47±3, 50±1,
and 51±1%, 500 mg/L, respectively), higher than ribavirin (38±1, 36±1, 38±1, and
36±1%, 500 mg/L, respectively). Thus, compound 22 is a promising candidate for
anti-TMV development. Most of these compounds show broad-spectrum fungicidal
activities against 14 kinds of phytopathogenic fungi and selectively fungicidal
activities against Physalospora piricola, Sclerotinia sclerotiorum, and Rhizoctonia
cerealis. Additionally, some of these compounds exhibit insecticidal activity against
Culex pipiens pallens, Mythimna separata, Helicoverpa armigera, and Pyrausta
nubilalis. Compound 17 exhibits the highest larvicidal activity (LC was 0.32 mg/L)
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against Culex pipiens pallens.
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KEYWORDS: spirooxindoles, hydantoin, thiohydantoin, urea, thiourea,
characterization, antiviral activity, fungicidal activity, insecticidal activity
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INTRODUCTION
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Natural products have proven to be an important source of novel compounds for
drug discovery.1,2 They posses many of the properties that make a good drug
candidates, including structural diversity, specificity, and novel modes of action.3
However, natural products also have some disadvantages such as high structural
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complexity, limited compound availability, and poor drug-likeness. Structure
modification on the core skeletons of natural products is a powerful method for
overcoming these limitations and improving the efficiency of drug discovery.5
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Spirooxindoles are a core skeleton widely existing in natural products and drugs
(Figure 1). In recent years, more and more spirooxindole natural products and their
derivatives have been reported to have new drug development value, and their
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application prospect has attracted wide attention. However, these research are
concentrated in the field of pharmaceuticals, few research have been done in
pesticides.7 β-Carbolines and spirooxindole alkaloids share a same biogenic
precursor−tryptophan.8, 9 In previous work, we found that β-carboline alkaloids and
tryptophan showed antiviral, fungicidal, and insecticidal activities.10, 11 So, we wonder
whether spirooxindole derivatives also have similar pesticide activities.
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Hydantoin, thiohydantoin, urea and thiourea are important privileged scaffolds
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that widely exist in natural products, medicines, and pesticides (Figure 1). Part of
these natural products and their derivatives exhibit wide spread biological activities,
which have not only been used as antitumor,13 anti-HIV,14 antiarrhythmic,15
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antiepileptic, and anxiolytic agents in medicinal chemistry but also been used as
pesticides.11, 18 Previous works has demonstrated that the introduction of these
structures is beneficial for improving the biological activities of compounds
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containing these scaffolds. These structures contain multiple hydrogen bond donors
and receptors, which increase the probability or intensity of the interactions between
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small organic molecules and target proteins. The inclusion of these structures into
spirooxindoles would increase the molecular hydrogen bonding sites thus might
improve their bioactivities.
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Based on the above points, a series of spirooxindole derivatives containing
hydantoin, thiohydantoin, urea, and thiourea moieties were designed, synthesized,
characterized and the diastereoselectivity mechanism were studied in detail. Their
anti-TMV, fungicidal, and insecticidal activities were evaluated for the first time . The
structure-activity relationship study of these new spirooxindole derivatives was also
discussed (Figure 2).
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MATERIALS AND METHODS
Instruments
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H, C nuclear magnetic resonance (NMR) spectra were recorded on Bruker
Avance 400 Ultrashield NMR spectrometers. Chemical shifts (δ) were given in parts
per million (ppm) and were measured downfield from internal tetramethylsilane.
High-resolution mass spectrometry (HRMS) data were obtained on an FTICR-MS
instrument with a q-TOF mass analyzer (Ionspec 7.0 T). Single-crystal X-ray
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structure data were collected on a Rigaku Saturn 70. The melting points were
determined on an X-4 microscope melting point apparatus and are uncorrected.
Conversion was monitored by thin layer chromatography (TLC). Flash column
chromatography was performed over silica gel (200-300 mesh).
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General Synthesis
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Ribavirin (Topscience Co., Ltd.), chlorothalonil (Bailing Agrochemical Co.,Ltd)
and other reagents were purchased from commercial sources and were used as
received. All anhydrous solvents were dried and purified according to standard
techniques just before use. The synthetic routes are given in Scheme 1.
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The operating steps and physical data in detail of intermediates A−E and target
compounds 1−27 can be found in the Supporting Information.
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Biological Assay
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Each bioassay was repeated three times at 25 ± 1 °C. Activity results were
estimated according to a percentage scale of 0-100 (0, no activity; 100, total kill). The
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anti-TMV, fungicidal, and insecticidal activities of the synthesized compounds
were tested using previously reported methods, The results are presented as means ±
standard errors. The detail biological assay methods were given in Supporting
Information.
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RESULTS AND DISCUSSION
Synthesis
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The synthesis of the key intermediate spirooxindole E is depicted in Scheme 1.
Using L-tryptophan as starting material, the tetrahydrocarboline carboxylic acid A
was constructed via Pictet−Spengler cyclization, which then reacted with thionyl
chloride and methanol to form methyl ester B. The obtained methyl ester B was
subsequently reacted with (Boc) O to afford compound C. The spirooxindole D was
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obtained via the spirocyclization reaction in the presence of N-bromosuccinimide
(NBS) and glacial acetic acid. It is worth mentioning that the cyclization reaction is
stereoselective. The NMR (nuclear magnetic resonance) spectra showed that the E
was a single isomer, indicating that the newly formed quaternary carbon was
chiral-specific. The possible mechanism is shown in Figure S7. In molecule C, the
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ester group is axially oriented, it overlaps the π orbital during the bromination step.
+
The intermediate H is formed with the partial charge of Br distribute to ester oxygen
atom, which leads to the formation of cis-3-bromo-indolenine I. The following
addition of water to the imine group requires a before hand conformational change of
ring C (J). The stereospecific cis-bromohydrin (K) has perfectly aligned bonds for the
subsequent pinacol-type rearrangement that offers the observed product D. The
spirooxindole E is formed from the spirooxindole D by deprotection. Compounds
1-15 containing the thiohydantoin fragment can be synthesized from the spirooxindole
E with the corresponding isothiocyanates and triethylamine (Scheme 1). In order to
explore the effects of substituent at 5′-position and chirality at 3- and 7a'-positions on
anti-TMV activities, 16 and 17 containing thiohydantoin were synthesized from L-and
D-tryptophan, respectively. L-and D-tryptophan went through similar reactions to
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form compounds F and G,7 respectively. Then, they reacted with phenyl
isothiocyanates to form 16 and 17 (Scheme 1). Spirooxindole analogues containing
hydantoin (18-20) were obtained by reacting E with the corresponding isocyanates
(Scheme 1). When E reacted with tert-butyl isothiocyanates or some isocyanates, only
uncyclized thiourea derivative 21 or urea derivatives 22-27 were obtained (Scheme 1).
The single crystal structure of compound 6 further confirmed the chiral configuration
of quaternary carbon (Figure S6 and Table S1).25
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Configuration.
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It is interesting that both thiohydantoin (1-15) and hydantoin (18-20) derivatives
exhibite two sets of peaks in NMR spectra (Figure S8). However, there is only one set
of peaks in the NMR spectra of thiourea (19), and urea (22-27) derivatives (Figure
S9). Since the chirality of the quaternary carbon and the chirality of carbon which
connected to the methyl formate is determined, both factors can be ruled out.
Therefore, the only factor orients the charity of nitrogen. As we know, the lone pair
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occupies one sp orbital of nitrogen atom. In the case of thiourea (19) and urea
(22-27) derivatives, the barrier to pyramidal inversion of nitrogen is not high, and the
rate of inversion in solution is too fast to be measured by NMR spectroscopy.
Therefore, there is only one set of peaks for thiourea (19) and urea (22-27) derivatives
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(Figure S10). When the fused ring has been produced, such as thiohydantoin (1-15)
and hydantoin (18-20) derivatives, nitrogen atom inversion is restricted due to the
high inversion barriers, thus a pair of invertomers are generated resulting in two set of
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peaks in NMR spectra (Figure S10). When there is a methyl substitution in the
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5'-position, the cyclization reaction is stereospecific due to steric hindrance, resulting
in a single chiral nitrogen (16-17).
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Antiviral Activities.
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In general, most of these spirooxindole analogues exhibit higher anti-TMV
activities in vitro and inactivation, curative, and protection effects in vivo than
compound E (Table 1), which suggests that the introduction of these privileged
scaffolds is indeed beneficial to antiviral activities. Part of the compounds, such as
compounds 4, 16, and 22, show significantly higher activities than that of
commercialized anti-plant virus agent ribavirin (38±1, 36±1, 38±1, and 36±1%, 500
mg/L).
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For derivatives containing thiohydantoin moiety (1-15): When the substituent on
the N of thiohydantoin is substituted phenyl (1-4), the electronic effect of the
substituents on the phenyl has effect on the anti-TMV activities. The
electron-donating substituent is beneficial to improving anti-TMV activities compared
with electron-withdrawing substituent. For example, the structure-activity relationship
shows as follows: o-methyl (4) > non-substituent (1) > m-chloro (2) ≈ p-chloro (3).
The activities decrease obviously when the phenyl is changed to aliphatic substituents
(5-8). The size of cyclic substituents also has an effect on the activities, expressed as
the compound containing cyclopentyl (6) exhibits lower activities than those of
cyclohexyl (5). But the chain length of the substituents has little effect on activities (7
and 8). It is adverse to the anti-TMV activities when phenyl is changed to benzyl
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(9-15). The activities of derivative with an electron-donating substituent on the benzyl
ring (15) are significantly better than that with electron-withdrawing substituents
(10-14). For example, the activities of derivative with a p-methoxyl substituent (15,
35±1, 31±1, 28±2, and 29±1%, 500 mg/L) are higher than that with a p-chloro
substituent (11, 0, 6±1, 0, and 0, 500 mg/L). The position of the substituents on the
benzyl also affect the activities, the structure-activity relationship shows as follows:
m-chloro substituent (12) ≈ o-chloro substituent (13) > dichloro substituent (14) >
p-chloro substituent (11). When the α position of N (5′ position) is substituted by a
methyl, compound 16 (44±1, 44±2, 48±1, and 50±1%, 500 mg/L) exhibits higher
activities than that of unsubstituted one (1), even higher than that of ribavirin. Chiral
has important influence on the anti-TMV activities, the activities of isomer 16 are
significantly better than that of isomer 17. Compounds containing hydantoin moiety
(18-20) exhibits similar activities to the corresponding compounds containing
thiohydantoin moiety.
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For the derivatives containing urea moiety, they have a similar structure-activity
relationship to derivatives containing thiohydantoin moiety: the activities of the
compound containing aromatic substituent on N (22) are better than that of
compounds containing aliphatic substituents (21, 23-27). Compound 22 shows the
highest activities (45±1, 47±3, 50±1, and 51±1%, 500 mg/L), which is significantly
higher than that of ribavirin (Figure S11). This compound (22) can be used as an
anti-TMV candidate for further study.
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Fungicidal Activities
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In general, these compounds have a broad spectrum of fungicidal activities
against 14 kinds of phytopathogenic fungi (Table 2). Most of the derivatives exhibit
better fungicidal activities than compound E. Almost all these compounds show
fungicidal activities selectively against Physalospora piricola, Sclerotinia
sclerotiorum, and Rhizoctonia cerealis. Among them, compounds 1, 8, and 12 show
more than 60% fungicidal activities against 6 fungi. In particular, compound 21 shows
more broad-spectrum fungicidal activities, with more than 60% fungicidal activities
against 9 fungi. Compounds 8, 18, 20, and 22 exhibit more than 90% against
Physalospora piricola at 50 mg/L, compounds 6 and 20 show 90% and 96%
fungicidal activities, respectively, against Sclerotinia sclerotiorum at 50 mg/L.
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Insecticidal Activities
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Most of these derivatives show insecticidal activities against lepidoptera pests
such as Mythimna separata, Helicoverpa armigera, and Pyrausta nubilalis (Table 3).
All compounds show larvicidal activity against mosquito (Culex pipiens pallens), and
most of these derivatives show higher larvicidal activity against mosquito than that of
compound E. Thiohydantoin, thiourea, and urea compounds exhibit higher activities
than those of hydantoin compounds (Table 3). Chiral also has an important influence
on the mosquito larvicidal activity, thus the activity of isomer 17 is significantly better
than that of isomer 16. Compound 17 shows 40% inhibitory effect against Culex
pipiens pallens at 0.5 mg/L, and its LC against Culex pipiens pallens is 0.32 mg/L
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(Table S2), whereas, compound 16 exhibits only 40% mosquito larvicidal activity at 5
mg/L.
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In summary, a series of novel spirooxindole derivatives containing hydantoin,
thiohydantoin, urea, and thiourea moieties were designed and synthesized based on
the widely used privileged scaffolds in drug design. Firstly, we proposed a
diastereoselectivity mechanism and analyzed the conformation. Then, we
systematically studied the biological activity of the new compounds. The bioassays
results showed that the target compounds possessed moderate to good activities
against tobacco mosaic virus (TMV), among which compound 22 showed the highest
antiviral activity both in vitro and in vivo. At the same time, most of these compounds
exhibited broad-spectrum fungicidal activities against 14 kinds of phytopathogenic
fungi and selectively fungicidal activities against Physalospora piricola, Sclerotinia
sclerotiorum, and Rhizoctonia cerealis. Additionally, some of these compounds
exhibited insecticidal activity to Culex pipiens pallens, Mythimna separata,
Helicoverpa armigera, and Pyrausta nubilalis. Further investigations on structural
optimization and mode of action are in progress in our laboratory.
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ASSOCIATED CONTENT
Supporting Information.
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The operating steps and physical data in detail of intermediates A−E and target
compounds 1−27. Crystal data and structure refinement of 6 (Table S1 and Figure
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S6). The diastereoselectivity mechanism (Figure S7). Copies of H NMR of 15 and 27
(Figure S8 and Figure S9). Explanation of the cause of the difference in NMR spectra
(Figure S10). Detailed bioassay methods for anti-TMV, fungicidal, and insecticidal
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activities. Pictures of inactivation of 22, ribavirin, and blank control against TMV
(Figure S11). LC value of compound 17 against C. pipiens pallens (Table S2). This
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material is available free of charge via the Internet at http://pubs.acs.org.
AUTHOR INFORMATION
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Corresponding Authors
*Tel: +86-(0)22-23503952. Fax: +86-(0)22-23503952.
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E-mail: songhongjian@nankai.edu.cn (H. J. S.); chemzhangjingjing@163.com (J. J.
Z.); wangqm@nankai.edu.cn (Q. M. W).
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Funding
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We are grateful to the National Natural Science Foundation of China (21977056,
21732002), the Fundamental Research Funds for the Central Universities, Nankai
University (63201043), and the Tianjin Technical Expert Project (19JCTPJC57800)
for generous financial support for our programs.
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Notes
The authors declare no competing financial interest.
REFERENCES
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(1) Newman, D. J.; Cragg, G. M. Natural products as sources of new drugs from 1981
to 2014. J. Nat. Prod. 2016, 79, 629–661.
2
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(2) Clardy, J.; Walsh, C. Lessons from natural molecules. Nature 2004, 432, 829–837.
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(3) Nicolaou, K. C.; Hale, C. R.; Nilewski, C.; Ioannidou, H. A. Constructing
molecular complexity and diversity: total synthesis of natural products of biological
and medicinal importance. Chem. Soc. Rev. 2012, 41, 5185−5238.
56
57
2
2
58
59
(4) Yao, H.; Liu, J.; Xu, S.; Zhu, Z.; Xu, J. The structural modification of natural
products for novel drug discovery. Expert Opin. Drug Dis. 2017, 12, 121−140.
2
2
60
61
(5) Wang, S. Z.; Dong, G. Q.; Sheng, C. Q. Structural Simplification of Natural
Products. Chem. Rev. 2019, 119, 4180−4220.
2
2
62
63
(6) Zhou, L.-M.; Qu, R.-Y.; Yang, G.-F. An overview of spirooxindole as a promising
scaffold for novel drug discovery. Expert Opi. Drug Dis. 2020, 15, 603−625.
2
2
2
2
64
65
66
67
(7) Chen, L. W.; Xie, J. L.; Song, H. J.; Liu, Y. X.; Gu, Y. C.; Wang, L. Z.; Wang, Q.
M. Design, synthesis, and biological activities of spirooxindoles containing
acylhydrazone fragment derivatives based on the biosynthesis of alkaloids derived
from tryptophan. J. Agric. Food Chem. 2016, 64, 6508−6516.
2
2
2
2
2
68
69
70
71
72
(8) Ding, Y. S.; de Wet, J. R.; Cavalcoli, J.; Li, S. Y.; Greshock, T. J.; Miller, K. A.;
Finefield, J. M.; Sunderhaus, J. D.; McAfoos, T. J.; Tsukamoto, S.; Williams, R. M.;
Sherman, D. H. Genome-based characterization of two prenylation steps in the
assembly of the stephacidin and notoamide anticancer agents in a marine-derived
Aspergillus sp. J. Am. Chem. Soc. 2010, 132, 12733–12740.
2
2
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(9) Berlin, J.; Rügenhagen, C.; Greidziak, N.; Kuzovkina, I.; Witte, L.; Wray, V.
Biosynthesis of serotonin and β-carboline alkaloids in hairy root cultures of Peganum
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Page 16 of 33
2
75
Harmala. Phytochemistry 1993, 33, 593–597.
2
2
2
2
76
77
78
79
(10) Song, H. J.; Liu, Y. X.; Liu, Y. X.; Wang, L. Z.; Wang, Q. M. Synthesis and
antiviral and fungicidal activity evaluation of β-carboline, dihydro-β-carboline,
tetrahydro-β-carboline alkaloids, and their derivatives. J. Agric. Food Chem. 2014,
62, 1010−1018.
2
2
2
2
80
81
82
83
(11) Huang, Y. Q.; Liu, Y. X.; Liu, Y. X.; Song, H. J.; Wang, Q. M. C ring may be
dispensable for β-carboline: Design, synthesis, and bioactivities evaluation of
tryptophan analog derivatives based on the biosynthesis of β-carboline alkaloids.
Bioorg. Med. Chem. 2016, 24, 462–473.
2
2
84
85
(12) Choa, S. H.; Kim, S.-H.; Shin, D. Y. Recent applications of hydantoin and
thiohydantoin in medicinal chemistry. Eur. J. Med. Chem. 2019, 164, 517–545.
2
2
2
86
87
88
(13) Jung, M. E.; Ouk, S.; Yoo, D.; Sawyers, C. L.; Chen, C.; Tran, C.; Wongvipat, J.
Structure-activity relationship for thiohydantoin androgen receptor antagonists for
castration-resistant prostate cancer (CRPC). J. Med. Chem. 2010, 53, 2779–2796.
2
2
2
2
2
89
90
91
92
93
(14) Romine, J. L.; St Laurent, D. R.; Leet, J. E.; Martin, S. W.; Serrano-Wu, M. H.;
Yang, F. K.; Gao, M.; O'Boyle, D. R.; Lemm, J. A.; Sun, J. H.; Nower, P. T.; Huang,
X. H.; Deshpande, M. S.; Meanwell, N. A.; Snyder, L. B. Inhibitors of HCV NS5A:
from iminothiazolidinones to symmetrical stilbenes. ACS Med. Chem. Lett. 2011, 2,
224–229.
2
94
(15) Matsukura, M.; Daiku, Y.; Ueda, K.; Tanaka, S.; Igarashi, T.; Minami, N.
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2
2
95
Synthesis
and
antiarrhythmic
activity
of
2,2-dialkyl-1′-(Nsubstituted
96
97
aminoalkyl)-spiro-(chroman-4,4′-imidazolidine)-2′,5′-diones. Chem. Pharm. Bull.
1992, 40, 1823–1827.
2
2
3
3
98
99
00
01
(16) Brouillette, W. J.; Jestkov, V. P.; Brown, M. L.; Akhtar, M. S.; DeLorey, T. M.;
Brown, G. B. Bicyclic hydantoins with a bridgehead nitrogen. Comparison of
anticonvulsant activities with binding to the neuronal voltage-dependent sodium
channel. J. Med. Chem. 1994, 37, 3289–3293.
3
3
3
3
3
02
03
04
05
06
(17) Lápez-Rodríguez, M. L.; Rosado, M. L.; Benhamú, B.; Morcilló, M. J.;
Fernández, E.; Schaper, K. J. Synthesis and structure–activity relationships of a new
model of arylpiperazines. 2. Three-dimensional quantitative structure-activity
relationships of hydantoin-phenylpiperazine derivatives with affinity for 5-HT1A and
α1 receptors. A comparison of CoMFA models. J. Med. Chem. 1997, 40, 1648–1656.
3
3
3
3
07
08
09
10
(18) Huang, Y. Q.; Guo, Z. L.; Song, H. J.; Liu, Y. X.; Wang, L. Z.; Wang, Q. M.
Design, synthesis, and biological activity of β-carboline analogues containing
hydantoin, thiohydantoin, and urea moieties. J. Agric. Food Chem. 2018, 66,
8253−8261.
3
3
3
3
11
12
13
14
(19) Lu, A. D.; Ma, Y. Y.; Wang, Z. W.; Zhou, Z. H.; Wang, Q. M. Application of
“Hydrogen-bonding interaction” in drug design. Part 2: Design, synthesis, and
structure–activity relationships of thiophosphoramide derivatives as novel antiviral
and antifungal agents. J. Agric. Food Chem. 2015, 63, 9435−9440.
3
15
(20) Lu, A. D.; Wang, Z. W.; Zhou, Z. H.; Chen, J. X.; Wang, Q. M. Application of
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3
16
17
“hydrogen bonding interaction” in new drug development: design, synthesis, antiviral
activity, and SARs of thiourea derivatives. J. Agri. Food Chem. 2015, 63, 1378−1384.
3
3
3
18
19
20
(21) Wang, Z. W.; Wei, P.; Xu, X. Z.; Liu, Y. X.; Wang, L. Z.; Wang, Q. M. Design,
synthesis, and antiviral activity evaluation of phenanthrenebased antofine derivatives.
J. Agric. Food Chem. 2012, 60, 8544−8551.
3
3
3
3
21
22
23
24
(22) Liu, Y. X.; Zhang, P. X.; Li, Y. Q.; Song, H. B.; Wang, Q. M. Design, synthesis,
and biological evaluation of 2-benzylpyrroles and 2-benzoylpyrroles based on
structures of insecticidal chlorfenapyr and natural pyrrolomycins. Mol. Divers. 2014,
18, 593−598.
3
3
3
3
25
26
27
28
(23) Ma, Q. Q.; Liu, Y. X.; Zhang, P. X.; Li, Y. Q.; Xiong, L. X.; Wang, Q. M.
Design, synthesis, and biological evaluation of various α-substituted benzylpyrroles
based on the structures of insecticidal chlorfenapyr and natural pyrrolomycins. J.
Agric. Food Chem. 2014, 62, 6072−6081.
3
3
29
30
(24) France, D. J.; Los, M. The direct formation of a cis-bromohydrin from an
8β-methyloestrane derivative. J. Chem Soc. D Chem. Commun. 1969, 24, 1513−1514.
3
3
3
31
32
33
(25) CCDC 1999602 contains the supplementary crystallographic data for this paper.
These data can be obtained free of charge from The Cambridge Crystallographic Data
Centre via www.ccdc.cam.ac.uk/data_request/cif.
3
3
34
35
(26) Dewar, M. J. S.; Jennings, W. B. Barrier to pyramidal inversion of nitrogen in
dibenzylmethylamine. J. Am. Chem. Soc. 1971, 93, 401–403.
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(27) Srivastava, A.; Srivastava, V.; Verma, S. M.; Subramanian, E. Restricted
Inversion of Pyramidal Nitrogen through Π -Electronic Interaction in an Acyclic
System. J. Org. Chem. 1994, 59, 3560–3563.
37
38
3
39
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40
41
3
3
42
43
Figure 1. Natural products and drugs containing spirooxindole, thiohydantoin,
hydantoin, thiourea, and urea scaffolds.
3
3
3
3
3
3
3
44
45
46
47
48
49
50
Figure 2. Design of target compounds.
Scheme 1. Synthesis of spirooxindole analogues 1-27.
3
3
3
51
52
53
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HO
N
O
O
N
H
H
N
N
H
H
O
N
OH
MeO
N
H
MeN
H
O
HO
O
O
F
N
H
N
O
N
H
H
Cl
N
Cl
MeO
O
H
(
-)-Horsfiline
Strychnofoline
Spirotryprostatin A
MI-147
O
O
O
F
O
O
HN
NH
S
H
N
S
N
N
O
NH
N
O
Cl
O
N
N
H
O
O
O
NH
Br
Cl
N
NH
OH
Macahydantoin B
H
OEt
Satavaptan
O
NITD609
Axinohydantoin
CF3
O
O
NC
Cl
O
F
O
F
O
S
NH
S
F
NH
O
O
N
N
N
HN
H
H
HN
N
O
S
HN
O
O
N
N
H
H
Phenytoin
Enzalutamide
Diflubenzuron
Thiophanate-methyl
3
3
54
55
56
Figure 1.
3
O
R
N
O
N
X
H
N
X
X
NH
O
N
O
O
N
N
H
NH
H
H
DESIGN
O
N
H
X
multiple hydrogen bond donors and receptors
core skeleton
N
HN
R
privileged scaffolds
N
O
H
X = O or S
3
3
57
58
Figure 2.
3
3
3
59
60
61
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O
OH
O
O
O
N
OH
O
(Boc) O
TEA
O
NBS
2
NH2
HCHO
Pictet-Spengler
SOCl2
AcOH
NH
NH
MeOH
Boc
DCM
THF-H O
N
N
H
N
H
N
H
2
H
L-tryptophan
A
B
C
R
N 3'
2
'
R = phenyl (1)
benzyl (9)
O
1
'
3
-chlorophenyl (2) benzo[d][1,3]dioxol-5-ylmethyl (10)
4-chlorophenyl (3) 4-chlorobenzyl (11)
S
7a'
N
4
'
m-tolyl (4)
3-chlorobenzyl (12)
2-chlorobenzyl (13)
3,4-dichlorobenzyl (14)
4-methoxybenzyl (15)
7'
6
3
'
cyclohexyl (5)
cyclopentyl (6)
heptyl (7)
5
O
'
2
N
H
1
butyl (8)
O
O
O
R
N
O
Boc
O
N
R-NCX
TEA
con.HCl
dioxane
NH
O
R = phenyl (18)
N
4
-chlorophenyl (19)
O
3-chlorophenyl (20)
O
DCM
N
H
N
O
(CCDC 1999602)
H
6
N
H
D
E
O
O
X
X = S, R = t-butyl (21)
R
N
X = O, R = m-tolyl (22)
cyclohexyl (23)
N
H
cyclopentyl (24)
O
25
butyl ( )
N
H
26
)
benzyl (27)
t-butyl (
O
OH
O
O
O
N
O
OH
S
N
NH2
CH3CHO
Ph-NCS
NH
NH
con.H2SO4
DCM TEA
N
N
H
O
H
O
L-tryptophan
N
N
H
H
F
16
O
OH
O
O
O
O
N
OH
NH2
CH CHO
3
Ph-NCS
S
N
NH
NH
con.H2SO4
N
H
DCM TEA
N
H
O
O
D-tryptophan
N
H
G
N
H
17
3
3
3
63
64
65
Scheme 1.
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Table 1. In Vitro and in Vivo Anti-TMV Activity of Synthesized Compounds 1−27.
inhibition rate (%)/500 mg/L
compd
structure
protection
curative
effect
in vitro inactivation
effect
O
N
N
1
2
3
4
5
6
7
8
9
31±1
20±1
29±1
35±1
16±1
24±1
25±2
20±2
12±1
41±1
27±1
21±1
HN
S
O
O
N
N
Cl
HN
S
O
O
Cl
N
N
HN
S
O
O
N
N
45±1
28±1
0
40±1
29±1
8±1
50±2
10±2
6±1
46±2
13±1
0
HN
S
O
O
N
S
N
HN
O
O
N
N
S
HN
O
O
N
N
10±1
12±1
21±1
13±1
15±2
13±1
18±1
26±3
20±1
21±1
14±2
17±1
10±2
8±1
S
HN
O
O
N
N
HN
S
O
O
N
N
28±1
31±2
HN
S
O
O
N
N
1
1
0
1
HN
S
O
O
O
O
N
S
N
HN
0
6±1
0
0
O
Cl
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O
N
1
1
2
3
N
30±1
23±1
16±1
13±2
23±1
17±1
10±2
9±1
HN
Cl
O
S
O
Cl
N
N
HN
S
O
O
N
HN
N
1
1
4
5
0
14±1
31±1
22±2
28±2
10±1
29±1
Cl
O
S
Cl
O
N
S
N
HN
35±1
O
O
O
1
1
1
1
2
6
7
8
9
0
N
44±1
19±1
37±1
25±1
20±1
44±2
27±1
30±1
36±1
33±2
48±1
32±3
17±1
31±2
21±1
50±1
24±1
33±3
28±1
22±1
HN
N
O
S
O
N
HN
N
N
O
S
O
N
HN
O
O
O
N
Cl
HN
N
O
O
O
N
HN
N
O
Cl
O
O
O
2
1
N
S
19±1
20±1
13±1
22±1
HN
HN
O
O
O
N
O
2
2
2
2
3
4
HN
45±1
0
47±3
0
50±1
8±1
51±1
0
O
HN
O
N
O
O
HN
HN
O
O
O
O
N
26±1
27±1
11±2
21±2
HN
HN
O
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Journal of Agricultural and Food Chemistry
O
O
O
N
2
2
2
5
6
7
13±1
36±1
31±1
31±2
36±2
42±2
15±1
29±1
34±1
30±1
32±2
43±2
HN
HN
O
O
O
O
N
HN
HN
O
O
O
O
N
HN
N
O
H
O
O
E
NH
9±1
10±1
36±1
15±1
38±1
22±1
36±1
HN
O
N
N
O
O
N
HO
NH2
ribavirin
38±1
HO
OH
3
67
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Table 2. Fungicidal Activity of Compounds 1−27 against Fourteen Kinds of Phytopathogens.
fungicidal activity (%)/50 mg/L
compd
F.Ca
54±1
14±1
18±3
23±1
18±1
46±2
C.H
63±3
25±1
31±1
38±1
50±2
56±1
P.P
A.S
F.G
F.M
S.S
P.C
R.C
B.M
W.A
64±2
32±3
56±2
40±1
36±1
56±1
P.I
R.S
B.C
1
2
3
4
5
6
76±1
70±2
76±1
80±2
72±1
72±1
58±2
26±2
37±2
37±1
37±2
47±1
56±1 46±2
86±3
80±2
70±1
70±1
53±1
90±1
59±1
41±1
45±1
24±2
37±1
44±3
84±1 56±1
40±1 30±1
56±1 26±1
62±1 30±3
62±1 22±3
73±1 52±1
42±1
16±1
5±1
54±1
12±1
12±1
10±1
12±1
30±1
70±2
42±1
56±1
50±1
33±1
81±1
28±1
8±2
28±1 15±1
13±2 15±1
15±1
5±1
33±1
8±1
23±2 31±1
27±2
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Journal of Agricultural and Food Chemistry
7
8
9
36±1
50±3
41±1
23±3
41±1
36±1
32±3
23±1
32±1
23±1
56±3
63±1
38±1
38±1
44±1
50±1
44±1
44±1
50±1
63±1
76±1
93±1
80±2
80±2
80±4
90±3
80±2
62±1
72±1
76±1
58±1
42±2
53±1
47±1
47±1
63±1
32±1
47±1
40±3
30±1
56±2 46±1
42±1 46±2
39±1 31±1
76±1
83±1
74±2
84±2
73±2
88±1
66±1
66±1
57±2
84±1
31±2
45±1
48±1
24±1
37±1
37±1
38±1
41±1
32±1
17±1
80±1 48±1
82±1 60±3
60±1 41±3
62±1 30±2
73±1 48±1
82±1 48±2
70±1 37±2
62±1 33±1
73±1 44±1
73±4 37±1
40±2
60±1
56±1
44±1
48±1
44±1
52±1
36±1
52±1
44±1
26±1
21±2
37±1
6±1
49±1
24±1
24±1
20±1
25±1
30±1
12±1
37±1
10±1
20±1
56±1
58±1
61±1
78±2
70±2
64±1
47±1
39±1
67±2
14±1
1
0
1
2
3
4
5
6
6±1
15±1
1
1
1
1
1
1
19±1 46±1
62±1 46±1
56±1 31±1
47±1 10±2
45±1 15±1
23±2 15±1
30±1
30±1
21±1
16±1
21±1
15±1
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1
1
1
2
2
2
2
2
2
2
7
8
9
0
1
2
3
4
5
6
18±2
23±1
9±1
25±1
56±1
25±1
75±3
13±1
63±2
0
72±2
93±1
60±3
90±3
62±2
90±2
70±1
45±1
62±2
35±1
32±1
23±1
26±1
63±1
16±1
47±1
10±1
16±1
16±1
30±1
36±1 15±1
23±1 23±1
22±1 15±1
70±1 54±1
72±2
77±1
72±2
96±2
36±1
60±1
10±1
20±1
33±1
20±1
52±1
12±1
28±1
42±1
31±1
17±2
7±1
71±2 33±1
64±1 44±1
40±1 19±1
84±1 63±1
49±2 19±1
62±1 33±1
47±2 15±1
40±1 11±2
47±1 19±1
38±2 11±1
44±1
48±1
32±2
70±2
32±1
44±3
36±1
36±1
16±1
28±1
26±2
15±1
16±1
46±2
11±1
15±1
12±1
11±2
16±1
6±1
22±1
25±1
12±1
20±1
12±1
13±1
15±1
12±1
15±1
10±1
42±1
33±1
14±3
75±1
14±1
42±1
17±1
11±1
17±1
28±1
50±1
14±1
27±1
9±1
42±1
0
23±1 31±1
6±2
7±1
14±2
14±1
5±2
19±1
6±1
19±1 15±1
14±1
10±1
12±1
28±1
17±1
8±1
0
13±1
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Journal of Agricultural and Food Chemistry
2
7
14±1
15±2
6±1
0
62±3
30±2
100
32±1
30±2
73±2
42±1 23±1
40±1 31±1
26±1
33±1
<50
17±1
10±1
100
50±2 33±1
44±1 35±1
36±1
30±1
91±1
16±1
12±1
86±1
17±1
15±1
100
14±2
14±1
100
E
chlorothalonil 100
73±2
<50
100
100
91±1
a
3
3
3
69
70
71
F.C: Fusarium oxysporiumf. sp.cucumeris; C.H: Cercospora arachidicola Hori; P.P: Physalospora piricola; A.S: Alternaria solani; F.G:
Fusarium graminearum; F.M: Fusarium moniliforme; S.S: Sclerotinia sclerotiorum; P.C: Phytophthora capsici; R.C: Rhizoctonia cerealis; B.M:
Bipolaris maydis; W.A: Watermelon-anthracnose; P.I: Phytophthora infestans; R.S: Rhizoctonia solani; B.C: Botrytis cinerea.
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