Design, Synthesis, and Insecticidal Activities of Novel Pyranoside Derivatives Targeting at Potential Second Calcium Channel IP3Receptor

Article abstract of DOI:10.1002/cjoc.201600460

In order to search for potent and environmental friendly insecticides with new modes of action, a series of pyranoside derivatives mimicking D-myo-inositol 1,4,5-trisphosphate (IP₃) were designed and synthesized according to the bioisosteric approach. The biological assay indicated that most of the new compounds showed moderate to good insecticidal activities against oriental armyworm (Mythimna separata) and diamondback moth (Plutella xylostella). Especially, compound 5g displayed 80% larvicidal activity against oriental armyworm at 50 mg/L. Meanwhile, 5a showed 100% and 70% larvicidal activities against diamondback moth at 50 and 25 mg/L, respectively. To further explore the mode of action, calcium imaging technique was applied to study the effects of 5a, 5g, and 5i on the intracellular calcium ion concentration ([Ca²⁺]_i) in central neurons isolated from Spodoptera exigua. The results indicated that the tested compounds released stored calcium ions from endoplasmic reticulum.

Full text of DOI:10.1002/cjoc.201600460

FULL PAPER
DOI: 10.1002/cjoc.201600460
Design, Synthesis, and Insecticidal Activities of Novel
Pyranoside Derivatives Targeting at Potential Second
Calcium Channel IP3 Receptor
Changchun Wu, Yuxin Li, Ming Liu, Jingbo Liu, Xuewen Hua, Dongkai Zhang, Lixia Xiong,
Na Yang, Baolei Wang, and Zhengming Li*
State Key Laboratory of Elemento-Organic Chemistry, Collaborative Innovation Center of Chemical Science and
Engineering (Tianjin), Nankai University, Tianjin 300071, China
In order to search for potent and environmental friendly insecticides with new modes of action, a series of
pyranoside derivatives mimicking D-myo-inositol 1,4,5-trisphosphate (IP3) were designed and synthesized
according to the bioisosteric approach. The biological assay indicated that most of the new compounds showed
moderate to good insecticidal activities against oriental armyworm (Mythimna separata) and diamondback moth
(Plutella xylostella). Especially, compound 5g displayed 80% larvicidal activity against oriental armyworm at 50
mg/L. Meanwhile, 5a showed 100% and 70% larvicidal activities against diamondback moth at 50 and 25 mg/L,
respectively. To further explore the mode of action, calcium imaging technique was applied to study the effects of
5a, 5g, and 5i on the intracellular calcium ion concentration ([Ca2＋]i) in central neurons isolated from Spodoptera
exigua. The results indicated that the tested compounds released stored calcium ions from endoplasmic reticulum.
Keywords pyranoside derivatives, insecticidal activity, IP3, bioisosteric approach
Introduction
In recent years, some conventional insecticides such
as methamidophos, dicofol, monocrotophos, fipronil,
etc. have been prohibited in China due to their toxicity
to mammals or damage to the proliferation of bees.
Therefore, the discovery of new potent insecticides with
novel modes of action is crucial to overcoming the re-
sistance and ecological problems. Anthranilic diamides
were discovered by DuPont as a new class of insecti-
cides targeting at the insect ryanodine receptors
(RyRs).[1,2] This category of insecticides has the intrinsic
selectivity for the insect RyRs without any unfavorable
effects towards mammals.[3] They can evoke massive
calcium release from intracellular stores, disrupt calci-
Figure 1
cyantraniliprole.
Chemical structures of chlorantraniliprole and
D-myo-Inositol 1,4,5-trisphosphate (IP3, Figure 2) is
an intracellular second messenger that stimulates mobi-
＋
lization of intracellular Ca2 stores.[9] When IP3 inter-
um homeostasis, and cause the final death of insects.[4,5]
＋
Ca2 release is mediated by intracellular ligand-
acts with a site on the N-terminal portion of its receptor,
＋
Ca2 ions are released from intracellular stores to the
gated calcium release channels, which have been identi-
fied as the ryanodine receptors (RyRs) and inositol
1,4,5-trisphosphate receptors (IP3Rs).[6] There are func-
tional similarities between RyRs and IP3Rs channels.[7,8]
Chlorantraniliprole and Cyantraniliprole (Figure 1) have
been marketed as environmentally-benign and highly
potent activators of insect RyRs. However, to the best of
our knowledge, little attention has been paid to discov-
ering new insecticides which target at the insect IP3Rs.
cytosol.[10,11] IP3 analogues have shown diverse biologi-
cal activities, such as anticancer and anti-inflammatory
activities in mammals study.[12-14] Adenophostins A and
B, as penicillium fermentated natural products, were
＋
reported to stimulate the release of Ca2
from
IP3-sensitive intracellular stores.[15,16] The adenophostins
were reported as pharmacological tools to investigate
cell signalling mechanisms of mammals.[17,18] They
*
E-mail: nkzml@vip.163.com; Tel.: 0086-022-23503732; Fax: 0086-022-23503732
Received July 22, 2016; accepted August 24, 2016; published online XXXX, 2016.
Supporting information for this article is available on the WWW under http://dx.doi.org/10.1002/cjoc.201600460 or from the author.
Chin. J. Chem. 2016, XX, 1—8
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Wu et al.
FULL PAPER
Synthesis of allyl 2,6-di-O-trimethylacetyl-3,4-bis-O-
(diethoxyphosphoryl)-α-D-glucopyranoside (5a)
were shown to be 10- to 100-fold more potent than IP3
＋
in releasing Ca2 and in receptor binding assays.[19]
A
crystal structure of the IP3-binding core of mouse IP3R1
in complex with IP3 was preliminarily confirmed by
X-ray technique,[10] and the delicate structure of insect
IP3Rs still remains unknown. Till today, there are no
reports on the study of new IP3Rs modulators of any
insects.
A solution of n-BuLi (1.9 mL, 4.8 mmol, 2.5 mol/L
in hexane) was added to a stirred mixture of allyl 2,6-
di-O-trimethylacetyl-α-D-glucopyranoside 1 (0.78 g, 2
mmol) in dry THF (15 mL) at 78 ℃ under nitrogen.
After reacting for 30 min at -78 ℃, diethyl chloro-
phosphate (1.04 g, 6 mmol) was added. The solution
was then warmed to room temperature gradually and
stirred overnight. The reaction was quenched with water
(20 mL), followed by extraction with EtOAc (30 mL×
3). The combined organic layers were washed with sat-
urated NaHCO3 (50 mL), brine (50 mL), and dried over
MgSO4. After filtration and evaporation in vacuo, the
obtained yellow crude residue was purified by column
chromatography on silica gel using petroleum
ether/EtOAc as eluent (V/V＝3∶1 then 1∶1) to afford
5a as a colorless liquid.
Considering a bioisosteric and mimicking approach,
inositol connected to a phosphate moiety in IP3 could be
replaced by a six member ring pyranoside. Inspired by
commercial RyRs insecticides, we first reported here the
design and synthesis of pyranoside derivatives contain-
ing phosphorus moieties as potential IP3Rs modulators.
To explore its mode of action, the effect of target com-
＋
pounds on [Ca2 ]i in the central neurons isolated from S.
exigua were studied by calcium imaging technique.[20]
Experimental
Synthesis of allyl 2,6-di-O-trimethylacetyl-3,4-bis-O-
(diphenoxyphosphoryl)-α-D-glucopyranoside (5b)
General
Intermediate 1 (1.16 g, 3 mmol) was dissolved in
anhydrous THF (20 mL) under nitrogen and cooled to
78 ℃. n-Butyllithium (2.9 mL, 7.2 mmol, 2.5 mol/L
in hexane) was then added, followed by addition of di-
phenyl chlorophosphate (1.1g, 9 mmol). After stirring
for 25 min at 78 ℃, the solution was warmed to room
temperature overnight. After completion of the reaction,
the mixture was poured into a suspension of diethyl
ether (60 mL) and water (60 mL), and extracted with
EtOAc (90 mL). The combined organic layers were
washed with saturated NaHCO3 (60 mL) and dried with
anhydrous MgSO4. The mixture was concentrated in
vacuo and purified by column chromatography on silica
gel using petroleum ether/EtOAc as eluent [V/V＝10∶1
with 2% Et3N] to afford 5b (1.15 g, 45%) as a yellow
liquid.
1H NMR, 13C NMR and 31P NMR spectra were rec-
orded on a Bruker AV 400 spectrometer in CDCl3 solu-
tion with tetramethylsilane as the internal standard.
High-resolution mass spectrometry (HRMS) data were
recorded on an Agilent 6520 Q-TOF LC/MS spectrom-
eter with ESI resource. The reagents used were all ana-
lytically or chemically pure. All solvents were dried by
standard methods in advance and distilled before use.
All yields referred to isolated yields and were not opti-
mized. Spots were visualized by spraying with phos-
phomolybdic acid (10% in EtOH) followed by heating
or by dipping into an aqueous basic solution of KMnO4
followed by heating. D-myo-Inositol 1,4,5-trisphosphate
hexasodium salt (IP3Na) was purchased from J&K Sci-
entific Ltd (Beijing, China).
H2N
N
N
HO
N
OR
N
OH
HO
O
O
H2O3PO
H2O3PO
H2O3PO
H2O3PO
OPO3H2
OH
O
OPO3H2
OH
IP3
Adenophostin A R = H
Adenophostin B R = Ac
OR2
OR2
OR1
OR1
O
R1O
O
OR2
OR1
R1O
S
O
P
O
R1O
R1O
O
P
S
P
O
O
O
O
O
O
OR1
OR2
P
OR2
P
P
OR1
OR2
OR3
OR3
R1O
OR3
OR1
R1O
Figure 2 Chemical structures of some representative compounds and design of the title compounds.
2
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Pyranoside Derivatives Targeting at Potential Second Calcium Channel IP3 Receptor
Synthesis of 1-O-allyl-2,6-di-O-trimethylacetyl-α-D-
was quenched with crushed ice. The organic layer was
washed with brine (30 mL), dried over MgSO4, and fil-
trated. Evaporation of the filtrate in vacuo gave the yel-
low residue, which was purified by column chromatog-
raphy on silica gel [petroleum ether/EtOAc 60∶1 (V∶
V) with 2% Et3N] to afford pure 5f as a colorless oil.
glucopyranosyl-3,4-bis-(dibenzylphosphite) (5c)
1H-Tetrazole (1.12 g, 16 mmol) was added portion-
wise to a stirred solution of 1 (1.56 g, 4 mmol) in dry
CH2Cl2 (30 mL) at room temperature under nitrogen.
Dibenzyl-N,N-diisopropylphosphoramidite (4 mL, 12
mmol) was then added. After stirring at room tempera-
ture overnight, the solution was concentrated in vacuo.
The residue was purified by column chromatography on
silica gel [petroleum ether/EtOAc (V∶V＝50∶1) with
2% Et3N] to yield 5c as a colorless oil.
Synthesis of allyl 2,6-di-O-trimethylacetyl-3,4-bis-
O-(diethoxythiophosphoryl)-α-D-glucopyranoside
(5g)
To a mixture of 1 (1.16 g, 3 mmol) and diisopro-
pylethylamine (1.94 g, 15 mmol) in dry CH2Cl2 (40 mL)
at 40 ℃ under nitrogen was added diethyl chloro-
phosphite (1.41 g, 9 mmol) dropwise. After reacting for
1 h at40 ℃, the mixture was warmed to room tem-
perature slowly and stirred for another 12 h. Then, a
solution of phenylacetyl disulfide (3.0 g, 10 mmol) in
CH2Cl2 was added. After 24 h, the mixture was diluted
with EtOAc (90 mL), washed with water. The organic
layer was dried over MgSO4, and concentrated under
reduced pressure to give the yellow residue, which was
purified by silica gel column chromatography [petro-
leum ether/EtOAc 50∶1 then 20∶1 (V∶V)] to give 5g
as a yellow liquid.
Synthesis of allyl 2,6-di-O-trimethylacetyl-3,4-bis-O-
(dibenzyloxyphosphoryl)-α-D-glucopyranoside (5d)
To a stirred solution of 1 (1.16 g, 3 mmol) and
1H-tetrazole (0.84 g, 12 mmol) in dry CH2Cl2 (15 mL)
at room temperature under nitrogen was added diben-
zyl-N,N-diisopropylphosphoramidite (3 mL, 9 mmol).
After reacting for 12 h, the mixture was then cooled to
−78 ℃, followed by addition of m-CPBA (2.43 g, 85%,
12 mmol) in portions over 10 min. The resulting mix-
ture was warmed to room temperature gradually, and
stirred overnight. The reaction was quenched with a
10% aqueous solution of Na2SO3 (25 mL). The organic
layer was separated, and the aqueous layer was extract-
ed with CH2Cl2. The combined organic layers were
washed with saturated NaHCO3 (120 mL), dried over
MgSO4, and concentrated. Purification of the residue by
column chromatography on silica gel [petroleum
ether/EtOAc 6∶1 then 3∶1 (V∶V)] gave 5d as a yel-
low liquid.
Synthesis of allyl 2,6-di-O-benzyl-3,4-bis-O-(dieth-
oxyphosphoryl)-α-D-glucopyranoside (5h)
The same procedure was followed as that described
above for title compound 5a, except for the use of allyl
2,6-di-O-benzyl-α-D-glucopyranoside 4 instead of in-
termediate 1. All other steps remained the same.
Synthesis of n-propyl 3,4-bis-O-(diethoxyphospho-
ryl)-α-D-glucopyranoside (5i)
Synthesis of allyl 2,6-di-O-trimethylacetyl-3,4-bis-
O-(dibenzyloxythiophosphoryl)-α-D-glucopyranoside
(5e)
To a solution of compound 5h (0.5 g, 0.8 mmol) in
methanol was added Pd-C (10%, 1.0 g) at room temper-
ature. The reaction was stirred under an atmospheric
pressure of hydrogen for 24 h. The resulting mixture
was then filtered through a pad of Celite. Evaporation of
the filtrate followed by column chromatography on sili-
ca gel [petroleum ether/EtOAc 10∶1 then 2∶1 (V∶V)]
gave 5i as a colorless liquid.
Compound 1 (0.78 g, 2 mmol) was dissolved in dry
CH2Cl2 (25 mL) under nitrogen, and 1H-tetrazole (0.42
g, 6 mmol) was added, followed by dibenzyl-N,N-
diisopropylphosphoramidite (1.9 mL, 5 mmol). After
stirring for 6 h at room temperature, a solution of phe-
nylacetyl disulfide (3.0 g, 10 mmol) in CH2Cl2 was
added. The resulting mixture was stirred for another 24
h. The solvent was removed under reduced pressure,
and the residue was partitioned with ethyl acetate (60
mL) and water (60 mL). The organic layer was washed
with brine, dried, and concentrated. The residue was
purified by column chromatography on silica gel [(pe-
troleum ether/EtOAc 50∶1 then 20∶1 (V∶V)] to af-
ford 5e as a yellow liquid.
Synthesis of allyl 2,6-di-O-benzyl-3,4-bis-O-(dieth-
oxythiophosphoryl)-α-D-glucopyranoside (5j)
The same procedure was used as that described
above for title compound 5g, except for the use of 4 as
the substrate instead of 1. All other steps remained the
same.
Biological assay
Synthesis of 1-O-allyl-2,6-di-O-trimethylacetyl-α-D-
glucopyranosyl-3,4-bis-(diethylphosphite) (5f)
Insecticidal activities against oriental armyworm
(Mythimna separata) and diamondback moth (Plutella
xylostella) were performed on test organisms reared in a
greenhouse. The bioassay was replicated at (25±1) ℃
according to statistical requirements. Assessments were
made on a dead/alive basis, and mortality rates were
corrected using Abbott's formula. The error of the ex-
periments was 5%. For comparative purposes, D-myo-
Intermediate 1 (1.56 g, 4 mmol) and diisopro-
pylethylamine (2.58 g, 20 mmol) were dissolved in dry
CH2Cl2 (40 mL) under nitrogen atmosphere. The result-
ing mixture was cooled to 40 ℃, and diethyl chloro-
phosphite (1.88 g, 12 mmol) was added dropwise. After
stirring for 30 min at 40 ℃, the solution was warmed
to room temperature and stirred overnight. The reaction
Chin. J. Chem. 2016, XX, 1—8
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3

Wu et al.
FULL PAPER
inositol 1,4,5-trisphosphate hexasodium salt (IP3Na)
was tested as positive control under the same condi-
tions.
α-D-glucopyranoside (4) were synthesized referring to
the previous reports as shown in Scheme 1.[24,25]
Scheme 1 Synthesis of intermediates 1 and 4
Larvicidal activity against oriental armyworm
OH
OPiv
i Pivaloyl chloride,
ii 2-methoxypropene,
PTSA, THF, r.t.
The larvicidal activities of compounds 5a－5j and
D-myo-inositol 1,4,5-trisphosphate hexasodium salt
were evaluated according to the leaf-dip method.[21]
Leaf disks were cut from fresh corn leaves and then
dipped into the test solution. After drying, the treated
leaf disks were placed individually into a glass-surface
vessel. Then, 10 third-instar oriental armyworm larvae
were put into each vessel containing dried treated leaf
disk. The symptoms of affected larvae were observed 1
d after treatment. Mortality rates were evaluated 3 d
after treatment. Each treatment was replicated three
times.
pyridine, 0 oC
O
O
HO
HO
HO
HO
OH
OPiv
OAll
OAll
1
OBn
OPiv
O
O
O
iii NaOH, MeOH, reflux
iv NaH, BnBr, DMF
O
O
O
OBn
OAll
OPiv
OAll
2
3
OBn
O
HO
HO
v 1N HCl/MeOH, r.t.
OBn
OAll
4
Larvicidal activity against diamondback moth
All = allyl, Piv = (CH3)3CCO, Bn = benzyl
The larvicidal activities of compounds 5a－5j and
D-myo-inositol 1,4,5-trisphosphate hexasodium salt
were tested by the leaf dip method.[22] At first, a solution
of each test sample in DMF (AR, purchased from Alfa
Aesar) at a concentration of 1000 mg/L was prepared
and then diluted to the required concentration with dis-
tilled water. Leaf disks were cut from fresh cabbage
leaves and then dipped into the test solution for 10 s.
After drying, the resulting leaf disks were placed indi-
vidually into glass tubes. Each disk was infested with 30
third-instar diamondback moth larvae. Percentage mor-
talities were evaluated 3 d after treatment. Each treat-
ment was performed three times.
With the diol intermediate 1 in hand, we focused our
attention on the preparation of the glycoside bisphos-
phate 5a (Table 1). Initially, 1 was treated with diethyl
chlorophosphate in the presence of K2CO3 or KOH in
acetone under reflux. However, the expected product 5a
was not detected (Entries 1－2). Then, 1 was reacted
with diethyl chlorophosphate in the presence of DMAP
at room temperature.[26] To our delight, the desired
product was isolated in 10% yield (Entry 3). In order to
increase the yield, the reaction was conducted in CH2Cl2
under reflux. To our disappointment, elevating the reac-
tion temperature merely led to some side rections (Entry
4). It was found that the exposure of 1 to n-BuLi/diethyl
chlorophosphate resulted in the formation of product 5a
in 65% yield (Entry 5).[27]
Calcium imaging
Calibration of the fluorescence signal was achieved
by using the method of Takahashi et al. with modifica-
tions.[23] Briefly, the attached neurons were rinsed in
standard physiological saline [(mmol/L): NaCl 150, KCl
4, MgCl2 2, CaCl2 2, HEPES 10, buffered to pH 7.0]
and then incubated in the dark for 45 min at 28 ℃ in
standard external saline containing the dye fluo-3 AM
(10 µmol/L). After dye loading, cells were again rinsed
in physiological saline twice. Calcium free extracellular
fluid had the following composition (mmol/L): NaCl
150, KCl 4, MgCl2 2, EGTA 2, Hepes 10, buffered to
pH 7.0. After dye loading, neurons were again rinsed in
standard physiological saline twice. Calcium ratio im-
aging studies were conducted using the imaging system
coupled to an inverted fluorescence microscope with a
Fluor 40× oil immersion objective (Olympus IX71).
Cells were excited at 488 nm and the 530 nm fluores-
cence emission acquired using a CCD (Image Pro-6.0).
Table 1 Optimization of the reaction conditions
Entry
Reaction condition
K2CO3/acetone/reflux
KOH/acetone/reflux
Yielda/%
no reaction
no reaction
10
1
2
3
4
5
DMAP/CH2Cl2/r.t./48 h
DMAP/CH2Cl2/reflux/12 h
n-Buli/THF/78 ℃ to r.t.
side reaction
65
a Isolated yield.
As illustrated in Scheme 2, 1 was subjected to the
optimized conditions (n-BuLi, THF, 78 ℃ to r.t.) with
diphenyl chlorophosphate to give 5b in 45% yield. The
reaction of 1 with dibenzyl-N,N-diisopropylphos-
phoramidite and 1H-tetrazole in CH2Cl2 furnished bis-
Results and Discussion
Chemistry
The key intermediates allyl 2,6-di-O-trimethylace-
tyl-α-D-glucopyranoside (1) and allyl 2,6-di-O-benzyl-
4
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Pyranoside Derivatives Targeting at Potential Second Calcium Channel IP3 Receptor
Scheme 2 Synthetic route of target compounds 5b－5g
Biological activities
phosphites 5c, which could be stored without decompo-
sition in a freezer (at －20 ℃) under nitrogen. 5c was
then oxidized with m-CPBA to give 5d in 32% overall
yield (2 steps).[28] Unfortunately, the desired oxidative
product was not obtained when sulphur (S8) was used as
the oxidant.[29] Further treatment of 5c with phenyla-
cetyl disulfide in CH2Cl2 provided the expected
bisphosphorothioates 5e. On the other hand, 1 reacted
with diethyl chlorophosphite in the presence of DIPEA
to give 5f in 50% yield.[30] Product 5f should be also
stored in a freezer under nitrogen. Thionation of 5f with
phenylacetyl disulfide in CH2Cl2 at room temperature
gave 5g in 48% overall yield (2 steps).
Larvicidal activity against oriental armyworm
The insecticidal activities of title compounds and IP3
against oriental armyworm were listed in Table 2, from
which we observed that IP3 displayed a 50% fatal rate at
200 mg/L. Furthermore, 5a and 5j showed 20% insecti-
cidal activity at 100 mg/L (200 mg/L, 100%). It was
worth noting that 5g showed excellent larvicidal activi-
ties even at 50 mg/L, which was far better than that of
IP3.
Table 2 Insecticidal activities of compounds 5a－5j and IP3
against oriental armyworm
Larvicidal activity/% at conc./(mg•L−1)
Compd.
Intermediate 4 was then smoothly converted to 5h
by exposure to n-BuLi/diethyl chlorophosphate in 52%
yield (Scheme 3). Further reaction of 5h with Pd/C un-
der a pressure of H2 provided 5i in 84% yield. Finally, 4
was treated with diethyl chlorophosphite in the presence
of DIPEA providing the corresponding bisphosphites,
which further underwent a one-pot thionation with phe-
nylacetyl disulfide to give 5j in 47% yield.
500
100
50
200
100
15
100
20
0
50
0
5a
0
5b
60
15
0
0
5c
60
30
0
0
5d
30
10
0
0
5e
Scheme 3 Synthetic route of target compounds 5h－5j
75
50
0
0
5f
100
60
100
40
100
0
5g
80
0
5h
60
20
0
0
5i
100
NTa
100
50
0
5j
20
0
IP3Na
0
a NT: not tested
The bioassay results indicated that the substituted
groups had a significant influence on the larvicidal ac-
tivities. Compared with 5a, 5b and 5d, it was observed
that the sequence of larvicidal activity was Et＞Bn＞Ph
in the phosphate moiety. The bioassay results of 5e and
Chin. J. Chem. 2016, XX, 1—8
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5

Wu et al.
FULL PAPER
phosphate moiety. However, the sequence of insecticidal
activity was Bn＞Et in the phosphite and thiophosphate
moieties (5c＞5f, 5e＞5g). Moreover, bisphosphites 5c
and 5f were more effective than the corresponding bis-
thiophosphates 5e and 5g, respectively. From Table 3,
we could also conclude that the substituents (such as
pivaloyl, Bn, H) in the various moieties at the C-2 and
C-6 positions of pyranoside ring had minimal impact on
biological activities. In general, the structures of the title
compounds had no significant effect on the larcidical
activity against diamondback moth.
5g also followed the above sequence. It was concluded
that bisphosphites 5c and 5f showed lower larvicidal
activities than bisphosphates 5d and 5a, respectively. In
addition, bisthiophosphates 5g and 5j showed higher
larvicidal activities than bisphosphates 5a and 5h. The
bioactivities of compounds 5a, 5h, and 5g, 5j indicated
that the sequence of larvicidal activity was trimethyla-
cetyl＞benzyl in the different substituents at the C-2 and
C-6 positions of the pyranoside ring. The hydroxyl
groups at the C-2 and C-6 positions of the pyranoside
bisphosphate 5i did not have a positive effect on the
larvicidal activities. Hence, it was concluded that the
trimethylacetyl and diethyoxylthiophosphoryl groups
connected to oxygen atoms on the pyranoside ring had a
significant effect on the larvicidal activities and were
key pharmacophores.
Table 3 Insecticidal activities of compounds 5a－5j and IP3
against diamondback moth
Larvicidal activity/% at conc./(mg•L−1)
Compd.
500
100
100
100
100
100
100
100
100
100
100
100
200
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
60
50
100
80
100
0
25
70
40
55
0
10
40
0
Figure 3 showed the biological symptoms of larvaes
affected by title compound 5g, IP3 and chlorantranilip-
role (Rynaxypyr) individually. Insects treated with 5g
5a
5b
5c
＋
and IP3 showed typical Ca2 ion efflux symptoms such
30
0
as insect body contraction, body shortening and feeding
cessation. The symptoms were similar to those of lar-
vaes treated with commercial Rynaxypyr.
5d
5e
100
100
100
100
100
100
100
70
85
70
80
90
85
80
20
45
0
0
20
0
5f
5g
20
50
40
60
0
5h
5i
0
0
5j
IP3Na
45
Electrophysiological recordings
To determine whether the title compounds could be
involved in calcium signaling transduction and the tar-
get of the novel compounds, we examine the effects on
the central neurons of S. exigua on the calcium homeo-
stasis with calcium imaging technique after the neurons
loading with fluo-3 AM.
＋
Figure 4 showed the change of [Ca2 ]i vs. recording
time when the neurons were treated with 5a, 5i, 5g and
Figure 3 Symptoms of third-instar larvae of oriental armyworm
treated by leaf dipping. (a) 5g, (b) IP3, (c) Rynaxypyr, (d)
Untreated.
chlorantraniliprole in the absence of extracellular cal-
cium. Application of compound 5 could increase the
＋
cytosolic calcium concentration. The peaks of [Ca2 ]i
Larvicidal activity against diamondback moth
Table 3 showed the insecticidal activity of compounds
5a－5j and IP3 against diamondback moth. On the
whole, most of the tested compounds displayed good to
excellent larvicidal activities at 50 mg/L. Particularly,
5a and 5c possessed 100% fatal rate at 50 mg/L, a little
higher than that of IP3Na (80%, 50 mg/L). It was noted
that 5a, 5c, and 5i exhibited moderate larvicidal activi-
ties at 25 mg/L. Additionally, compounds 5a－5j and
IP3Na showed low insecticidal activity at 10 mg/L. For
example, the larvicidal activities of 5a and IP3Na at 10
mg/L were 40% and 45%, respectively.
were elevated to (104.2±1.37)% (n ＝ 6),
(104.7±2.1)% (n ＝ 6), (106.2±2.7)% (n ＝ 6) and
(111.8±3.5)% (n＝6) of the initial value after treat-
ments of the neurons with high concentration of 5a, 5i,
5g and chlorantraniliprole, respectively.
＋
Figure 5 showed the change of [Ca2 ]i vs. recording
time when the neurons were treated with 5g (100
mg•L−1), IP3Na (500 mg•L−1) and chlorantraniliprole (1
mg•L−1) in the absence of extracellular calcium. The
experiment results showed that lower chlorantraniliprole
could increase the free calcium and compound 5g could
not. On the other hand, after the neurons were treated
＋
with IP3Na for 2.5 min, the peaks of [Ca2 ]i were ele-
The bioactivities of 5a, 5b, and 5d revealed that the
sequence of larvicidal activity was Et＞Ph＞Bn in the
vated to (112.2±3.10)% (n＝6) and two peaks were
6
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© 2016 SIOC, CAS, Shanghai, & WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Chin. J. Chem. 2016, XX, 1—8

Pyranoside Derivatives Targeting at Potential Second Calcium Channel IP3 Receptor
observed through the recording. Although this novel
compound could cause the claium to be elevated in the
neurons, the concentration was too high.
be a new lead of potential IP3Rs modulators. Further
structural optimization is currently undertaken in our
laboratory.
Acknowledgement
This work was financially supported by the National
Natural Science Foundation of China (No. 31370039),
973 Program (No. 2010CB126106), the Tianjin Natural
Science Foundation (No. 16JCYBJC29400) and the
“111” Project of Ministry of Education of China (Pro-
ject No. B06005).
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Conclusions
In summary, ten novel compounds were designed,
synthesized and evaluated for their insecticidal activities
against oriental armyworm and diamondback moth. The
structures were characterized by 1H NMR, 13C NMR,
31P NMR, and HRMS. The bioassays showed that most
compounds exhibited good insecticidal activities. In
particular, compound 5g had better larvicidal effects
against oriental armyworm than IP3. Moreover, com-
pound 5a exhibited the same larvicidal level as IP3
against diamondback moth. The symptoms of larvaes
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Chin. J. Chem. 2016, XX, 1—8