CN113367144A

CN113367144A - Compound preparation of N6- (2-hydroxyethyl) adenosine and chlorantraniliprole

Info

Publication number: CN113367144A
Application number: CN202110641730.XA
Authority: CN
Inventors: 方鸣; 厉晓腊; 陈官菊; 刘又高; 柴一秋; 蔡瑞杭
Original assignee: ZHEJIANG SUBTROPICAL CROPS RESEARCH INSTITUTE
Current assignee: ZHEJIANG SUBTROPICAL CROPS RESEARCH INSTITUTE
Priority date: 2021-06-09
Filing date: 2021-06-09
Publication date: 2021-09-10

Abstract

The compound preparation of N6- (2-hydroxyethyl) adenosine and chlorantraniliprole comprises chlorantraniliprole and N6- (2-hydroxyethyl) adenosine. The synergistic mechanism between the two may be related to the continuous loss of calcium ions in cells. The generation of drug resistance of pests can be delayed by mixing chlorantraniliprole and HEA. The two can play a significant role in synergism after being mixed.

Description

Compound preparation of N6- (2-hydroxyethyl) adenosine and chlorantraniliprole

Technical Field

The invention relates to the field of pesticides, in particular to a compound preparation of N6- (2-hydroxyethyl) adenosine and chlorantraniliprole.

Background

Chlorantraniliprole (Chlorantraniliprole) is the first broad-spectrum insecticide with a novel chemical structure of anthranilamide benzamide, and has the characteristics of high efficiency on insects, low toxicity on mammals and high selectivity. The product is developed and created by DuPont company in the United states in 2008 and is successfully commercialized, and is widely favored by the market due to the characteristics of high efficiency and low toxicity. According to the application of Zhejiang plant protection departments in paddy fields for preventing and controlling rice leaf rollers, 10 milliliters of the pesticide is used per mu, and two barrels of the pesticide are sprayed by a knapsack manual sprayer and the pesticide is sprayed by a conventional method. The insecticidal effect reaches 94.2 percent after seven days, and the leaf protecting effect reaches 90.0 percent; the insecticidal effect reaches 86.0% after fourteen days, and the leaf protecting effect reaches 83.9%. Chlorantraniliprole can effectively control almost all important lepidoptera pests and partial other pests, and can also effectively control pests generating resistance to other pesticides. It has excellent quick acting and lasting effect and better crop protecting effect. However, after 2011, because of the difference between the frequency of medication and the rationality of medication, the resistance of diamondback moths to chlorantraniliprole is different in different regions and different years, and the diamondback moths have high-level resistance to the chlorantraniliprole in part of regions. The resistance of diamondback moths to chlorantraniliprole is reported to be increased to different degrees in the areas of Hainan, Guangzhou, Zhejiang and the like, the resistance index is different from 50 to 100 times, and the resistance level is high. At present, farmers in Wenzhou areas are few in number and even do not use chlorantraniliprole for control. At present, the compounding of chlorantraniliprole and gibberellin is applied to the field. The research of medicament compounding has become a necessary means for reducing the medicament dosage, improving the efficiency and delaying the resistance of chemical pesticides.

Disclosure of Invention

In order to solve the problems, the compound preparation of the N6- (2-hydroxyethyl) adenosine and the chlorantraniliprole is provided, which improves the drug effect of the chlorantraniliprole and reduces the application use amount.

In order to achieve the above purpose, the invention is solved by the following technical scheme: the compound preparation of N6- (2-hydroxyethyl) adenosine and chlorantraniliprole comprises chlorantraniliprole and N6- (2-hydroxyethyl) adenosine.

Further, the concentration of chlorantraniliprole is 0.808 mg/L.

Further, the concentration of N6- (2-hydroxyethyl) adenosine was 600 mg/L.

The invention has the advantages that:

the chlorantraniliprole has a good insecticidal effect on diamondback moth larvae, the extracted and separated active substance HEA (N6- (2-hydroxyethyl) adenosine) of cordyceps cicadae has a certain insecticidal effect on the diamondback moth larvae, but the efficiency of the insecticide on the market is far less, the effect that the HEA can effectively increase the semilethal concentration of the chlorantraniliprole can be seen through a compound synergy experiment, and the synergistic effect of the HEA and the chlorantraniliprole can have a certain relation with the action principle of the HEA and the chlorantraniliprole. The chemical structure of chlorantraniliprole has a brand-new insecticidal principle which is not possessed by any other insecticides, so that the chlorantraniliprole can efficiently activate an insect ryanodine (muscle) receptor to excessively release calcium ions in a calcium reservoir in cells, and the insects die by paralysis. HEA is a calcium ion antagonist which can selectively block calcium ions from flowing into cells through calcium ion channels on cell membranes, and reduce the concentration of calcium ions in the cells. The synergistic mechanism between the two may be related to the continuous loss of calcium ions in cells. The generation of drug resistance of pests can be delayed by mixing chlorantraniliprole and HEA. The two can play a significant role in synergism after being mixed.

Drawings

FIG. 1 is a schematic diagram of the mortality rate corrected by each treatment agent for 24h diamondback moth second instar larvae.

FIG. 2 is a schematic diagram of the mortality rate corrected by each treatment agent for the second instar larvae of diamondback moth in 48 h.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to fig. 1-2 in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments.

The test materials were as follows: diamondback moth 2-instar larva, 200 g/l chlorantraniliprole (DuPont, USA), 5% emamectin benzoate (pilot plant for agricultural chemicals in Profibus of Chinese academy of agricultural sciences), HEA (isolated and extracted from plant protection institute of subtropical crop institute), DMSO (dimethyl sulfoxide, purchased from Sigma Co.)

Virulence determination was performed by the leaf dipping method. Setting 5 concentration gradients of HEA, chlorantraniliprole medicament and emamectin benzoate, calculating a toxicity regression equation, and calculating an LC50 value. 200 g/L chlorantraniliprole and 5% emamectin benzoate are respectively prepared into semilethal concentrations of 0.808 mg/L and 0.9875 mg/L according to a virulence regression equation, HEA is firstly dissolved in 1% DMSO to be prepared into a concentration of 0.6mg/ml, and mass HEA with a concentration of 0.6mg/ml is dissolved in chlorantraniliprole with a concentration of LC50 and emamectin benzoate with a concentration of LC50 to be prepared into a compound preparation which is placed in a beaker for standby. Immersing fresh pollution-free cabbage leaves in each test solution for about 10s, taking out, naturally drying, putting into a culture dish, transferring 2-year-old diamondback moth larvae, and placing in an insect breeding chamber for constant-temperature breeding. Each treatment was repeated 3 times, and each 30 replicates were tested against 1% DMSO in water. And (5) checking the death condition of the test insects at 24h and 48h, touching the larva bodies with the brush pen tips, and determining that the test insects die without response. And calculating the mortality rate, correcting the mortality rate, and performing data processing by using DPS software to solve a virulence regression equation and LC 50. Data statistics is carried out by using SPSS venison 17 software, multiple groups of samples are subjected to difference significance analysis by a Duncan method, and graph making is carried out by using Graphpad Prism 8.0 software.

Influence of low temperature on age of diamondback moth larvae

Feeding temperature

Egg

Age 1

Age 2

Age 3

Age 4

Pupa

Imago

25℃

3.2±0.07

2.2±0.10

1.2±0.10

0.8±0.09

0.9±0.10

2.5±0.10

4.5±0.20

4℃

6.5±0.09

4.1±0.08

3.2±0.07

1.5±0.1

1.4±0.2

4.1±0.2

4.7±0.10

TABLE 1 calendar phase (d) of feeding diamondback moth larvae and pupae with artificial feed at different temperatures

As can be seen from Table 1, the growth cycle of the diamondback moth is increased under the low temperature condition, and the normal development of the diamondback moth is not influenced. At 25 ℃, the diamondback moth larvae take about 7-10 days to finish the period from 1 st to adult oviposition, and at low temperature, the diamondback moth larvae take about 20 days. Therefore, the larvae with different ages can be subjected to low-temperature preservation, and the population number required by the experiment can be obtained for subsequent experimental treatment. Compared with the method for feeding the radish seedling plutella xylostella, the artificial feed greatly saves manpower, material resources and space.

Indoor toxicity determination of three agents on diamondback moth 2-instar larvae

Reagent for testing	Regression equation of virulence	Correlation coefficient	LC50（mg/L）	95% confidence interval (mg/L)
					HEA	Y=-16.716+8.1447x	0.973904	463.7145	344.2400~624.6570
Chlorantraniliprole	Y=-5.2742+2.9684x	0.9908	0.80834	0.65471~1.5478
					Emamectin benzoate	Y=-5.0120+2.1203x	0.9878	0.9875	0.7143~1.4275

TABLE 2 virulence determination of diamondback moth larvae by three agents (48 h)

As can be seen from Table 2, the regression equations for the virulence of 200 g/L chlorantraniliprole and 5% emamectin benzoate are Y = -5.2742+2.9684x and Y = -5.0120+2.1203x, respectively, and the semilethal dose concentrations are 0.80834 mg/L and 0.9875 mg/L. The virulence regression equation Y = -16.716+8.1447x for HEA, with a semi-lethal dose concentration of 463.7145, indicating that HEA 600mg/L is the optimal insecticidal concentration and within the 95% confidence interval. Therefore, the concentration of HEA 600mg/L, the chlorantraniliprole medicament with the concentration of LC50 and the emamectin benzoate with the concentration of LC50 are selected for medicament compounding experiments.

Fig. 1 and 2 are indoor toxicity assays for HEA on chlorantraniliprole potentiation.

As can be seen from fig. 1, the mortality corrected for 24 hours after drug administration of the groups HEA (0.6 mg/ml), LC50 chlorantraniliprole, LC50 chlorantraniliprole + HEA (0.6 mg/ml), LC50 emamectin benzoate and LC50 emamectin benzoate + HEA (0.6 mg/ml) was 35.08%, 38.59%, 38.59%, 31.58%, 29.82%, respectively, and the mortality corrected for the group LC50 emamectin benzoate + HEA (0.6 mg/ml) was significantly lower than that of the other treatment group (P < 0.05), and there was no significant difference between the other four groups of treatments. Has no obvious synergistic effect 24 hours after being applied.

As shown in FIG. 2, the 48-hour corrected mortality rates of HEA (0.6 mg/ml), LC50 chlorantraniliprole, LC50 chlorantraniliprole + HEA (0.6 mg/ml), LC50 emamectin benzoate and LC50 emamectin benzoate + HEA (0.6 mg/ml) were 49.12%, 71.68%, 82.45%, 71.92% and 52.63%, respectively. The LC50 chlorantraniliprole + HEA (0.6 mg/ml) compound drug group is significantly higher than the single-dose HEA (0.6 mg/ml) group and the LC50 chlorantraniliprole group (P < 0.05), and the LC50 emamectin benzoate + HEA (0.6 mg/ml) compound drug group is significantly lower than the single-dose LC50 emamectin benzoate group (P < 0.05), and has no significant difference with the HEA (0.6 mg/ml) group.

The chlorantraniliprole and the emamectin benzoate have good insecticidal effects on diamondback moth larvae, the extracted and separated active substance HEA of cordyceps sobolifera also has a certain insecticidal effect on the diamondback moth larvae, but no insecticide on the market is highly efficient, and the compound synergy experiment shows that the HEA can effectively increase the action effect of the half-lethal concentration of the chlorantraniliprole, but the compound with the emamectin benzoate does not have synergy, but the toxicity is reduced. It is seen that not all lepidopteran insecticides can be compounded with HEA to enhance efficacy. The synergistic effect of HEA and chlorantraniliprole probably has a certain relation with the action principle of HEA and chlorantraniliprole. The chemical structure of chlorantraniliprole has a brand new insecticidal principle which any other insecticides do not have, and the chlorantraniliprole can efficiently activate receptors of the animalia spinosa (muscle) [8,9 ]. Excessive release of calcium ions from intracellular calcium stores leads to paralytic death of insects [10 ]. HEA is a calcium ion antagonist which can selectively block calcium ions from flowing into cells through calcium ion channels on cell membranes, and reduce the concentration of calcium ions in the cells. The synergistic mechanism between the two may be related to the continuous loss of calcium ions in cells.

The generation of drug resistance of pests can be delayed by mixing chlorantraniliprole and HEA. The research selects diamondback moth, a widely-occurring worldwide pest, as a target, researches the synergistic effect of the mixing of chlorantraniliprole and HEA by an indoor toxicity measurement method, and finds that the two can play a significant synergistic effect after being mixed.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof, and it is therefore intended that the present embodiments be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

Claims

A pharmaceutical formulation of N6- (2-hydroxyethyl) adenosine with chlorantraniliprole, wherein: including chlorantraniliprole and N6- (2-hydroxyethyl) adenosine.
2. The compound medicament of N6- (2-hydroxyethyl) adenosine with chlorantraniliprole according to claim 1, wherein: the concentration of chlorantraniliprole is 0.808 mg/L.
3. The compound medicament of N6- (2-hydroxyethyl) adenosine with chlorantraniliprole according to claim 2, wherein: the concentration of N6- (2-hydroxyethyl) adenosine was 600 mg/L.