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

Article abstract of DOI:10.1021/acs.jafc.5b02676

On the basis of the structure of natural product harmine, lead compound 18, and the structure of compounds in part 1, a series of thiophosphoramide derivatives 1-17 were designed and synthesized from various amines in one step. Their antiviral and antifungal activities were evaluated. Most of the compounds showed significantly higher antiviral activity against tobacco mosaic virus (TMV) than commercial virucide ribavirin. Compound (R,R)-17 showed the best anti-TMV activity in vitro (70%/500 μg/mL and 33%/100 μg/mL) and in vivo (inactivation effect, 68%/500 μg/mL and 30%/100 μg/mL; curative effect, 64%/500 μg/mL and 31%/100 μg/mL; protection effect, 66%/500 μg/mL and 31%/100 μg/mL), which is higher than that of ningnanmycin and lead compound 18. The antiviral activity of (R,R)-17·HCl is about similar to that of (R,R)-17. However, the antifungal activity of (R,R)-17·HCl against Puccinia sorghi is slightly lower than that of (R,R)-17. The systematic study provides compelling evidence that these simple thiophosphoramide compounds could become efficient antiviral and antifungal agents.

Full text of DOI:10.1021/acs.jafc.5b02676

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Article
Application of “hydrogen bonding interaction” in drug design
Part #: Design, synthesis, and SARs of thiophosphoramide
derivatives as novel antiviral and antifungal agents
Aidang Lu, Yuanyuan Ma, Ziwen Wang, Zhenghong Zhou, and Qingmin Wang
J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jafc.5b02676 • Publication Date (Web): 20 Oct 2015
Downloaded from http://pubs.acs.org on October 25, 2015
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Journal of Agricultural and Food Chemistry
Application of “hydrogen bonding interaction” in drug design Part Ⅱ:
Design, synthesis, and SARs of thiophosphoramide derivatives as novel
antiviral and antifungal agents
,
Aidang Lu†,*, Yuanyuan Ma†, Ziwen Wang‡ *, Zhenghong Zhou§, Qingmin Wang§,*
†School of Marine Science and Engineering, Hebei University of Technology, Tianjin 300130, China
‡Tianjin Key Laboratory of Structure and Performance for Functional Molecules, Key Laboratory of
InorganicꢀOrganic Hybrid Functional Material Chemistry, Ministry of Education, College of
Chemistry, Tianjin Normal University, Tianjin 300387, China
§State Key Laboratory of ElementoꢀOrganic Chemistry, Research Institute of ElementoꢀOrganic
Chemistry, Nankai University, Tianjin 300071, China
* To whom correspondence should be addressed. For Aidang Lu, Eꢀmail: luaidang@hebut.edu.cn;
Phone: 0086ꢀ22ꢀ60302241 Ext 205; Fax: 0086ꢀ22ꢀ60204274; For Ziwen Wang, Eꢀmail:
wangziwen2725@163.com; Phone: 0086ꢀ22ꢀ23503792; Fax: 0086ꢀ22ꢀ23503792; For Prof. Qingmin
Wang, Eꢀmail: wangqm@nankai.edu.cn; Phone: 0086ꢀ22ꢀ23503952; Fax: 0086ꢀ22ꢀ23503952.
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ABSTRACT: Based on the structure of natural product harmine, lead compound 18 and
the structure of compounds in part I, a series of thiophosphoramide derivatives 1–17
were designed and synthesized from various amines in one step. Their antiviral and
antifungal activities were evaluated. Most of the compounds showed significantly higher
antiviral activity against tobacco mosaic virus (TMV) than commercial virucide
Ribavirin. Compound (R,R)ꢀ17 showed the best antiꢀTMV activity in vitro (70%/500
ꢁg/mL and 33%/100 ꢁg/mL) and in vivo (inactivation effect: 68%/500 ꢁg/mL and
30%/100 ꢁg/mL; curative effect: 64%/500 ꢁg/mL and 31%/100 ꢁg/mL; protection effect:
66%/500 ꢁg/mL and 31%/100 ꢁg/mL) which is higher than that of Ningnanmycin and
lead compound 18. The antiviral activity of (R,R)ꢀ17—HCl is about similar to that of
(R,R)ꢀ17. However, the antifungal activity of (R,R)ꢀ17—HCl against Puccinia sorghi is
slightly lower than that of (R,R)ꢀ17. The systematic study provides compelling evidence
that these simple thiophosphoramide compounds could become efficient antiviral and
antifungal agents.
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KEYWORDS: tobacco mosaic virus, thiophosphoramide derivatives, antiꢀTMV activity,
Puccinia sorghi, antifungal activity, SARs
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INTRODUCTION
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Plant pathogens, including bacteria, fungi and viruses, result in substantial losses for
agriculture, and are considered a threat for food security. Bacteria can infect all plant
tissues during the whole plant growth cycle. More importantly, these bacteria have a
strong vitality. The infective capacity of phytopathogenic fungi can be maintained over
many seasons, so it is costly and difficult to for farmers control these fungal diseases.1 In
certain years, pathogenic fungi have reduced the yield of food and cash crops by nearly
twenty percent. Tobacco mosaic virus (TMV) is a wellꢀstudied plant virus which can
cause plant disease and creates enormous economic loss worldwide. There are about 36
plant families and more than 400 individual species which can be infected by TMV,
including tobacco and some ornamental flowers. Since plants do not have the same
immune system as animals, the control of plant diseases is still a challenge.2
Phosphoramides and their ester derivatives possess a wide range of biological
activities, such as pesticidal activity, antiviral activity, herbicidal activity, and so on.2–4
Several phosphoramides have become commercial varieties, such as acephate,
cyclophosphamide, methamidophos, and so on.
Antiꢀplant virus agent research is not like pharmaceutical research. Only very few
molecular targets are investigated and can be used in agrochemical design and discovery,
5
which increases the difficulty of discovery of antiviral molecules for plants. Hydrogen
bonding interaction is an important mode of action for small molecule and
biomacromolecule. In drug design and development, the applications of hydrogen
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bonding take more and more attention.6–8 Thiophosphoramide derivatives serve as
catalysts through hydrogen bonding from the NH group.9–13 Meanwhile, the TMV
composing primarily with coat protein and RNA, contains many amino acid residues.14, 15
If the thiophosphoramide derivatives can bind to the key residues of TMV CP or RNA,
they may interfere with virus assembly initiation.
In our previous work, we have been devoted to synthesizing βꢀCarboline and
tetrahydroꢀβꢀcarboline alkaloids as well as their derivatives and researching their
biological activity. Natural product harmine (Figure 1), one of the βꢀCarboline alkaloids
isolated from P. harmala L., was found to have excellent antiꢀTMV activity (antiviral
activity against TMV in vitro and inactivation, curative and protection activity in vivo
were 45%, 41%, 39%, and 42%, respectively, at 500 µg mL−1). Meanwhile,
(1S,3S)ꢀ1ꢀmethylꢀ2,3,4,9ꢀtetrahydroꢀ1Hꢀpyrido[3,4ꢀb]indoleꢀ3ꢀcarbohydrazide (Figure 1,
Lead compound 18), maintained high antiꢀTMV activity (the corresponding activity were
50%, 46%, 48%, and 50%, respectively, at 500 µg mL−1). This compound also exhibited
excellent stability and solubility.16 These advantages made it an ideal lead compound for
further derivation. A series of simple thiourea derivatives (Figure 1, Part I) were
developed as a novel type antiviral agents based on the structure characteristics of
harmine and lead compound 18.8
Taking into account the above findings, another series of simple thiophosphoramide
derivatives (Figure 1) were designed based on harmine, lead compound 18 and the
structure of compounds in part I and synthesized as antiꢀTMV agents. The synthesized
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compounds were characterized and systematically tested for their antiviral activity
against TMV and antifungal activity against Puccinia sorghi.
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MATERIALS AND METHODS
Instruments. The melting points of the synthesized compounds were determined on an
Xꢀ4 binocular microscope (Gongyi Yuhua Instrument Co., China). NMR spectra were
acquired via a Bruker 400 MHz (100 MHz for 13C) instrument at room temperature.
Chemical shifts were measured relative to residual solvent peaks of CDCl3 (1H: δ = 7.26
ppm; 13C: δ = 77.0 ppm) or d6ꢀDMSO (1H: δ = 2.50 ppm; 13C: δ = 39.5 ppm) with
tetramethylsilane as internal standards. HRMS data were obtained via an FTꢀICR MS
spectrometer (Ionspec, 7.0 T). Specific rotations were measured on a PerkinꢀElmer
341MC polarimeter. Symbol ( ±) represented transꢀmixture.
General Experimental Procedures
General Procedures for the Preparation of Compounds 1–3, 14 and (S,S)ꢀ17.
To a mixture of CH2Cl2 (10 mL), Et3N (5.0 mmol) and amine (5.0 mmol) was added
the corresponding diphenylphosphinothioic chloride (5.0 mmol or 0.5 mmol) in CH2Cl2
(10–30 mL), and stirred at room temperature. When the reaction was complete, the
reaction mixture was filtered or evaporated in vacuum. The pure products 1–3, 14 and
(S,S)ꢀ17 were obtained by recrystallization from diethyl ether or purified by
chromatography on a column of silica gel with PEꢀEtOAc (v/v = 3: 1).
NꢀButylꢀP,Pꢀdiphenylphosphinothioic amide (1): white solid, 99% yield; mp 90−91 °C;
1H NMR (CDCl3, 400 MHz): δ 0.89 (t, J = 7.2 Hz, 3H), 1.31−1.41 (m, 2H), 1.52−1.59
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(m, 2H), 2.23 (br. s, 1H), 2.92 (q, J = 7.2 Hz, 2H), 7.41−7.51 (m, 6H), 7.97−8.03 (m, 4H);
31P NMR (CDCl3, 161.7 MHz): δ 59.24; 13C NMR (CDCl3, 100.6 MHz): 13.8, 20.1, 33.6
(d, JCꢀP = 9.7 Hz), 128.5 (d, JCꢀP = 12.9 Hz), 131.5, 131.7, 133.4 (d, JCꢀP = 101.8 Hz);
HRMS (ESI) m/z calc'd for C16H20NPS [M +H]+: 290.1127, found 290.1124.
NꢀBenzylꢀP,Pꢀdiphenylphosphinothioic amide (2): white solid, 99% yield; mp
1
108−109 °C; H NMR (CDCl3, 400 MHz): δ 2.64 (br. s, 1H), 4.09 (d, J = 7.6 Hz, 2H),
31
7.24−7.37 (m, 5H), 7.44−7.50 (m, 6H), 8.01−8.06 (m, 4H); P NMR (CDCl3, 161.7
13
MHz): δ 59.82; C NMR (CDCl3, 100.6 MHz): 45.2, 127.6, 128.1, 128.5, 128.7 (d, JCꢀP
= 5.4 Hz), 131.7 (d, JCꢀP = 10.9 Hz), 131.9, 133.9 (d, JCꢀP = 101.8 Hz), 139.3 (d, JCꢀP
28.8 Hz); HRMS (ESI) m/z calc'd for C19H18NPS [M+H]+: 324.0970, found 324.0967.
=
NꢀCyclohexylꢀP,Pꢀdiphenylphosphinothioic amide (3): yellow solid, 99% yield; mp
1
77
−
80 °C; H NMR (CDCl3, 400 MHz): δ 1.09
−
1.29 (m, 5H), 1.52 1.66 (m, 3H),
−
2.03
−
2.06 (m, 2H), 2.27 (br. s, 1 H), 3.12 (s, 1H), 7.43
−
7.47 (m, 6H), 7.97−8.02 (m, 4H);
31P NMR (CDCl3, 161.7 MHz): δ 56.88; 13C NMR (CDCl3, 100.6 MHz): 25.2, 25.5, 36.1
(d, JCꢀP = 4.9 Hz), 51.1, 128.4 (d, JCꢀP = 12.9 Hz), 131.5, 131.6, 135.1 (d, JCꢀP = 101.9 Hz);
HRMS (ESI) m/z calc'd for C18H22NPS [M+H]+: 316.1283, found 316.1285.
Nꢀ(2ꢀAminoethyl)ꢀP,Pꢀdiphenylphosphinothioic amide (14): yellow solid, 95% yield;
mp 94
−
96 °C; 1H NMR (CDCl3, 400 MHz): δ 1.62 (s, 2H), 1.69 (br. s, 2H), 2.84
−
2.88 (m,
2H), 2.93
−
2.98 (m, 2H), 3.16 (br. s, 1 H), 7.42−4.07 (m, 6H), 7.96−
8.00 (m, 4H); 31P
13
NMR (CDCl3, 161.7 MHz): δ 59.78; C NMR (CDCl3, 100.6 MHz): 42.4 (d, JCꢀP = 8.4
Hz), 43.6, 128.5 (d, JCꢀP = 12.7 Hz), 131.6 (d, JCꢀP = 11.1 Hz), 131.7, 134.2 (d, JCꢀP
=
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102.0 Hz); HRMS (ESI) m/z calc'd for C14H17N2PS [M +H]+: 277.0923, found 277.0922.
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Nꢀ((1S,2S)ꢀ2ꢀAminoꢀ1,2ꢀdiphenylethyl)ꢀP,Pꢀdiphenylphosphinothioic
amide
((S,S)ꢀ17), 80% yield, [α]2D4 +21.7 (c 1.0, CHCl3), m.p. 103
−
105 °C; H NMR (CDCl3, 400
1
MHz): δ 1.60 (s, 2 H), 4.21 (t, J = 7.6 Hz, 1 H), 4.27 (d, J = 5.2 Hz, 1 H), 4.49−4.54 (m,
31
1 H), 7.11
−
7.18 (m, 5 H), 7.21
−
7.41 (m, 11 H), 7.62
−
7.70 (m, 4 H); P NMR (CDCl3,
13
161.7 MHz): δ 58.98; C NMR (CDCl3, 100.6 MHz): 61.2 (d, J = 7.4 Hz), 61.7, 126.9,
127.1, 127.3, 127.4, 127.7, 127.8, 127.9, 128.0, 128.1, 128.2, 131.0, 131.1, 131.2, 131.3,
131.7, 131.8, 133.5, 134.5, 134.7, 140.9, 142.5; HRMS (ESI) m/z calc'd for C26H25N2PS
[M +H]+: 429.1549, found 429.1547.
Procedures for the Preparation of Compound (R,R)ꢀ17—HCl
To a solution of 1.0 mL of HCl in ethyl acetate (2 M) and 10 mL of CH2Cl2 was
added 1.0 mmol of (R,R)ꢀ1711 at rt. The mixture was vigorously stirred for 6 h, and then
filtered to obtain the compound (R,R)ꢀ17—HCl.
Nꢀ((1R,2R)ꢀ2ꢀAminoꢀ1,2ꢀdiphenylethyl)ꢀP,Pꢀdiphenylphosphinothioic
hydrochloride ((R,R)ꢀ17—HCl): white solid, 99% yield; mp 155–158 °C; H NMR
(CDCl3, 400 MHz): δ 4.77 (br. s, 1 H), 4.97 5.04 (m, 1 H), 6.88 (t, J = 8.0 Hz, 1 H),
7.34 (m, 8 H), 7.43 7.45 (m, 2 H), 7.50 7.60 (m, 3 H),
7.79 (m, 2 H), 8.76 (s, 3 H); 31P NMR (CDCl3, 161.7 MHz): δ 57.61; 13C NMR
amide
1
−
6.97
−
−
7.11 (m, 5 H), 7.23
−
−
−
7.74
(CDCl3, 100.6 MHz): 59.3, 59.4 (d, J = 19.7 Hz), 127.0, 127.5, 127.6, 127.7, 128.0,
128.03, 128.11, 128.17, 128.26, 128.31, 130.8, 130.9, 131.0, 134.8, 135.0, 135.3, 135.8,
136.0, 139.1; HRMS (ESI) m/z calc'd for C26H26ClN2PS [M+H−HCl]+: 429.1549, found
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429.1548.
Biological Assay.
The antiviral and antifungal bioassays were performed via representative test organisms
reared in the laboratory. Percentage mortalities were evaluated according to a percentage
scale of 0−100, in which 0 indicates no activity and 100 indicates total kill.
Antiviral Biological Assay. The bioassays of in vitro and in vivo antiꢀTMV activity were
carried out on Nicotiana tabacum L. The detailed procedure of purifying TMV and the
method to test the antiꢀTMV activity of the synthesized compounds were the same with
the literature.17
Antifungal Biological Assay. The detailed assay method to test the antifungal activity of
the synthesized compounds was described in the literature.18
RESULTS AND DISCUSSION
Chemistry.
Compounds 1−3 , 14 and (S,S)ꢀ17 were synthesized according to procedures in Figure
2. Amines were reacted with corresponding phosphinothioic chloride19 to give
thiophosphoramides 1−3, 14 and (S,S)ꢀ17 in almost quantitative to moderate yields.
Compounds 4–13, 15, 16 and (R,R)ꢀ17 (Figure 3) were obtained according to our
9–12
previously reported methods.
HCl in good yields.
Compound 17—HCl was obtained in the presence of
Phytotoxic Activity. Firstly, compounds 1−17 were measured for their phytotoxic
activity against tobacco.17 The data of phytotoxic activity at 500 ꢁg/mL indicated that all
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of the compounds 1−17 showed no toxicity to the tested plant.
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Antiviral Activity. Compounds 1−17 were evaluated for their antiviral activity against
TMV. The commercial plant virucides Ningnanmycin, Ribavirin and lead compound 18
were used as the controls.
In Vitro Anti-TMV Activity. The results of antiviral assay in vitro were shown in Table 1.
As the control, Ribavirin exhibited a 41% inhibitory effect at 500 ꢁg/mL, whereas almost
all of the compounds 1−17 exhibited higher antiviral activity than Ribavirin even at the
concentration of 100 ꢁg/mL. The optimal compound (R,R)ꢀ17 exhibited significantly
higher inhibitory effect than Ningnanmycin, which had a good chance of becoming a
new lead compound for antiꢀTMV study. The ethylenediamine skeleton displayed
important role for keeping high antiviral activity. Thiophosphoramides 1−3 without
ethylenediamine skeleton exhibited relatively lower inhibitory effect than compounds
4−17 except for compound 14 (inhibitory effect: 3 ≈ 14). Among the compounds 7−10,
thiophosphoramide 9 displayed the lowest inhibitory effect, which indicated that alkyl
phosphate is bad for antiviral activity. The mainly difference between 11 and 12 lies in
the substituent on the N atom. Thiophosphoramide 11 with stronger hydrogen donor
(three NꢀH bond) showed higher inhibitory effect (inhibitory effect: 11 > 12)
.
Thiophosphoramide 11 displayed higher antiviral activity than 13, which indicated that
the S atom is favorable.
Diphenylthiophosphoramide derivatives 4, 11, 14−17 with a primary amine group
displayed in vitro activity ranging from 33%−70% against TMV at 500 ꢁg/mL. Amongst
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them, (R,R)ꢀ17 bearing 1,2ꢀ diphenylethylenediamine skeleton showed higher activity
than 11 containing 1,2ꢀcyclohexanediamine (70% and 62%, respectively). Except for
compound 14, all of primary amineꢀthiophosphoramide 7−17 exhibited much higher
activity than Ribavirin. The above results provided compelling evidence that a primary
amine on a rigid scaffold is necessary in thiophosphoramide molecule.
Compounds 17 exhibited best result with inhibitory rate of >60%. The optical pure
compounds 17 were prepared to evaluate the effect of configuration on antiviral activity.
(R,R)ꢀConfiguration was proven to be the optimal antiviral configuration in vitro
(inhibitory effect: (R,R)ꢀ17 > (S,S)ꢀ17). However, the activity of (S,S)ꢀ17 is about similar
to that of the mixture of (R,R)ꢀ17 and (S,S)ꢀ17 ((R,R)ꢀ17:(S,S)ꢀ17 = 1:1). In order to
improve stability and water solubility, (R,R)ꢀ17—HCl was synthesized in the presence of
HCl. However, the result indicated that (R,R)ꢀ17—HCl exhibited about similar in vitro
activity with (R,R)ꢀ17.
Bioassay results of the compounds 1−17 were shown in Table 1. Almost all of primary
amineꢀthiophosphoramide displayed excellent antiꢀTMV activity in vitro. Encouraged by
these results, the compounds 1−17 were tested to evaluate their antiviral activity in vivo.
In Vivo Anti-TMV Activity. The results of antiꢀTMV activity in vivo were shown in
Table 2. Most of the compounds exhibited higher in vivo activity than that of the
Ribavirin. At the concentration of 500 ꢁg/mL, (R,R)ꢀ17 displayed the best inhibitory
effect of inactivation activity, curative activity and protection activity with values of 68%,
64% and 66%, respectively. Ribavirin as a control was studied at the same conditions
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with values of 37%, 36% and 39%, respectively. By contrasting the experimental data,
the results indicated that compound (R,R)ꢀ17 was more efficient than Ribavirin in vivo
activity against TMV. Under the same test conditions, Ningnanmycin as a control was
also studied with values of 63%, 54% and 64%, respectively. At the concentration of 100
ꢁg/mL, (R,R)ꢀ17 displayed moderate activity (inactivation activity: 30%; curative
activity: 31%; and protection activity: 31%), which is obviously higher than that of
Ribavirin (inactivation activity: 9%; curative activity: 13%; and protection activity: 17%)
and Ningnanmycin (inactivation activity: 28%; curative activity: 22%; and protection
activity: 26%). (R,R)ꢀ17—HCl showed about similar in vivo antiꢀTMV activity with
(R,R)ꢀ17.
The in vivo antivral activity trend is similar with in vitro. Thiophosphoramide 11
bearing cyclohexanediamine skeleton gave relatively higher activity (64%, 60%, and
65% at 500 ꢁg/mL) than 1−10 and 12−13. Primary amineꢀthiophosphoramide 14−17
showed moderate antiviral activity ranging from 38%−71% against TMV at 500 ꢁg/mL
(the
antiviral
activity
in
vitro:
17
>16
>
15
>
14).
Nꢀ(2ꢀaminoethyl)ꢀP,Pꢀdiphenylphosphinothioic amide (14) showed relatively lower
activity (inactivation activity: 44%/500 ꢁg/mL and 11%/100 ꢁg/mL; curative activity:
40%/500 ꢁg/mL and 14%/100 ꢁg/mL; and protection activity: 38%/500 ꢁg/mL and
0%/100 ꢁg/mL) than corresponding primary amineꢀthiophosphoramides and higher than
1−3, which indicated that a primary amine on a rigid scaffold is necessary in
thiophosphoramide molecule for maintaining high antiviral activity.
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(R,R)ꢀConfiguration with higher activity than that of (S,S)ꢀconfiguration is confirmed
to be the optimal configuration (inhibitory effect: (R,R)ꢀ17 > (S,S)ꢀ17). (S,S)ꢀ17 showed
relatively lower activity (inactivation activity: 63%/500 ꢁg/mL and 30%/100 ꢁg/mL;
curative activity: 61%/500 ꢁg/mL and 20%/100 ꢁg/mL; and protection activity: 65%/500
ꢁg/mL and 26%/100 ꢁg/mL) than (R,R)ꢀ17.
In Vivo Fungicidal Activity. All the target compounds 1−17 were tested for their
fungicidal activity. Puccinia sorghi which causes great damage to corn was chosen as the
test object. Table 3 showed the results of inhibitory effect at different concentration
against Puccinia sorghi on corn in vivo. Compounds 1 and 17 (inhibitory effect at 200
mg/kg: ≥ 85%) exhibited higher fungicidal activity against Puccinia sorghi on corn than
compounds 2−16, thus emerged as new antifungal lead compounds. Compound (R,R)ꢀ17
exhibited the best fungicidal activity with 85% inhibitory effect at 100 mg/kg.
In summary, based on harmine, lead compound 18 and compounds in part I, a series of
simple thiophosphoramide compounds 1−17 were designed and synthesized from amines
in one step. Most of the compounds showed higher antiꢀTMV activity than commercial
virucide Ribavirin. Compound (R,R)ꢀ17 displayed the best antiꢀTMV activity in vitro and
in vivo (in vitro activity: 70%/500 ꢁg/mL and 33%/100 ꢁg/mL; inactivation activity:
68%/500 ꢁg/mL and 30%/100 ꢁg/mL; curative activity: 64%/500 ꢁg/mL and 31%/100
ꢁg/mL; protection activity: 66%/500 ꢁg/mL and 31%/100 ꢁg/mL) which is higher than
that of Ningnanmycin and lead compound 18. The antiviral activity of (R,R)ꢀ17—HCl is
about similar to that of (R,R)ꢀ17. However, the antifungal activity of (R,R)ꢀ17—HCl
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against Puccinia sorghi was slightly lower than that of (R,R)ꢀ17. Present studies on
antiviral and antifungal activity provide abundant support for further optimization of
these simple thiophosphoramide derivatives as new potential inhibitors of plant virus.
Further studies of these thiophosphoramide derivatives on structureꢀ activity relationship
and mode of action are in process in our laboratories.
ASSOCIATED CONTENT
Supporting Information
1H and 13C NMR spectra of compounds 1−3, 14 and 17. This material is available free of
charge via the Internet at http://pubs.acs.org.
AUTHOR INFORMATION
Corresponding Authors
*(A.L.) Eꢀmail: luaidang@hebut.edu.cn; Phone: 0086ꢀ22ꢀ60302241 Ext 205; Fax:
0086ꢀ22ꢀ60204274
*(Z.W.) Eꢀmail: wangziwen2725@163.com; Phone: 0086ꢀ22ꢀ23503792; Fax:
0086ꢀ22ꢀ23503792.
*(Q.W.) Eꢀmail: wangqm@nankai.edu.cn. Phone: 0086ꢀ22ꢀ23503952. Fax:
0086ꢀ22ꢀ23503952
Funding
This study was supported by National Natural Science Fundation of China (21302038,
21402142, 21132003), the Natural Science Fundation of Hebei Province (B2013202237),
the Program for Changjiang Scholars and Innovative Research Team in University
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(PCSIRT, IRT1059) and the Tianjin Natural Science Foundation (15JCQNJC05600),
Research Funds for College Students of Modern Marine Chemical Engineering
Technology Synergy Innovation Center of Hebei Province (HYHG201517).
Notes
The authors declare no competing financial interest.
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Figure Captions
Figure 1. Design of Thiophosphoramide Derivatives 1−17
Figure 2. Synthesis of Thiophosphoramide Derivatives 1–3, 14 and (S,S)ꢀ17
Figure 3. Chemical Structures of 4–13, 15, 16, (R,R)ꢀ17 and (R,R)ꢀ17·HCl
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Table 1. In Vitro Antiviral Activity of Compounds Ribavirin, Ningnanmycin and 1−17
against TMV
Concn
(ꢁg/mL)
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
Inhibition
rate (%)a
27±1
0
Concn
(ꢁg/mL)
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
Inhibition
rate (%)a
50±2
16±1
33±1
0
Compd
compd
13
1
2
31±2
0
14
34±1
0
55±1
27±1
49±2
0
3
15
55±2
24±1
50±1
24±1
55±2
20±1
63±1
28±2
60±2
32±2
47±1
15±1
58±2
28±1
62±2
32±2
50±1
22±2
4
16
70±1
33±2
62±2
24±1
60±2
25±1
72±2
36±1
50±2
21±1
45±2
20±1
41±2
10±1
61±1
26±1
5
(R,R)ꢀ17
(S,S)ꢀ17
6
(R,R)ꢀ17：
7
(S,S)ꢀ17 = 1: 1
8
(R,R)ꢀ17·HCl
Lead
9
compound 18
10
11
12
Harmine
Ribavirin
Ningnanmycin
338
339
340
aAverage of three replicates; All results are expressed as mean ± SD.
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Table 2. In Vivo Antiviral Activity of Compounds Ribavirin, Ningnanmycin and 1−17
against TMV
Concn
Inactivation
Curative
Protection
Concn
Inactivation
Curative
Protection
Compd
Compd
13
(ꢁg/mL)
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
effect (%)a effect (%)a effect (%)a
(ꢁg/mL) effect (%)a
effect (%)a effect (%)a
22±1
0
31±2
0
36±1
0
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
500
100
56±2
25±1
44±2
11±2
52±1
17±2
49±1
13±2
68±2
30±2
63±2
30±1
64±2
32±1
71±2
39±1
46±2
17±1
41±1
11±1
37±2
9±1
53±1
20±1
40±2
14±2
53±1
17±2
43±1
19±2
64±2
31±2
61±1
20±2
60±1
29±2
68±1
35±2
48±2
20±2
39±1
16±2
36±2
13±2
54±1
22±1
49±2
12±1
38±1
0
1
2
35±1
0
28±3
0
37±1
0
14
26±2
0
32±1
0
22±2
0
46±1
20±2
44±2
15±2
66±1
31±2
65±2
26±1
65±2
34±1
67±1
30±2
50±1
23±1
42±2
16±1
39±1
17±1
64±2
26±1
3
15
56±1
25±1
56±1
15±1
58±2
28±1
61±2
21±1
64±1
22±2
52±1
23±1
62±2
21±2
64±1
31±1
55±1
23±1
54±2
27±1
54±3
26±1
52±1
19±1
59±2
30±1
62±1
26±2
51±1
20±1
59±1
24±1
60±2
20±1
53±3
16±1
54±1
27±2
58±1
27±1
54±2
22±1
62±1
25±1
58±1
21±1
53±2
23±1
57±2
21±1
65±1
26±1
47±1
18±1
4
16
5
(R,R)ꢀ17
(S,S)ꢀ17
6
(R,R)ꢀ17：
7
(S,S)ꢀ17 = 1: 1
8
(R,R)ꢀ17·HCl
Lead
9
compound 18
10
11
12
Harmine
Ribavirin
63±2
28±1
Ningnanmycin
345
346
347
a Average of three replicates; All results are expressed as mean ± SD.
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Table 3. In Vivo Fungicidal Activity of the Selected Compounds against Puccinia sorghi
on corn
compd
Inhibition ratea(%)
200
85±1
70±2
40±3
50±1
45±2
40±3
30±1
35±2
30±3
20±1
50±2
60±2
40±3
20±1
30±3
10±1
98±2
95±1
98±2
100±1
100
75±2
NT
50
20±2
NT
Concn (mg/kg
)
1
2
3
NT
NT
4
NT
NT
5
NT
NT
6
NT
NT
7
NT
NT
8
NT
NT
9
NT
NT
10
NT
NT
11
NT
NT
12
13
NT
NT
NT
NT
14
NT
NT
15
NT
NT
16
NT
NT
(R,R)ꢀ17
(S,S)ꢀ17
(R,R)ꢀ17·HCl
Azoxystrobin
85±1
70±2
60±2
100±1
30±2
20±1
10±1
99±1
353
354
355
356
a Average of three replicates; All results are expressed as mean ± SD.
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Figure 1.
360
361
362
Figure 2.
363
364
365
366
367
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369
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Figure 3.
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380
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TOC graphic
Agrochemical Bioregulators
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