Application of amide compound as pesticide synergist

Abstract

The invention belongs to the technical field of pesticide synergists, and discloses application of an amide compound as a pesticide synergist. According to the invention, prodenia litura CYP6AE48 enzyme is taken as a main target, and a method of combining computer-aided drug design technology screening and biological effect screening is adopted, so that an amide compound, namely N-{5-[2-(2-{3-[2-(2-methylphenoxy) ethyoxyl] benzyl} hydrazino)-2-oxoethyl]-1, 3, 4-thiadiazole-2-yl} benzamide, is obtained through screening, namely N-{5-[2-(2-{3-[2-(2-methylphenoxy) ethyoxyl] benzyl} hydrazino)-2-oxoethyl]-1, 3, 4-thiadiazole-2-yl} benzamide. Tests show that the compound has a good inhibition effect on CYP6AE48 enzyme of prodenia litura, can prevent the prodenia litura from degrading lambda-cyhalothrin and chlorantraniliprole, and can be used as a pesticide synergist to play a synergistic role in pest control.

Description

Technical Field

[0001] The invention belongs to the technical field of pest control, and in particular relates to the use of an amide compound as a pesticide synergist. Background Art

[0002] Spodoptera litura Fabricius is a polyphagous and voracious pest belonging to the family Noctuidae of the order Lepidoptera. It has a strong reproductive capacity and can reproduce multiple generations within a year with overlapping generations. It can harm 389 species of host plants in 109 families. It is one of the main pests of vegetables, cotton, grains and oils, and various green plants.

[0003] At present, field pests are mainly controlled by chemical pesticides. This method is simple and convenient, can quickly and effectively control the expansion of pest populations, and has achieved remarkable control results. However, due to the long-term and repeated use of pesticides, the resistance of Spodoptera litura to pesticides has developed rapidly. For example, it has developed different levels of resistance to various pesticides such as cyantraniliprole, cypermethrin, and avermectin. Studies have shown that the mechanisms by which insects develop resistance include: reduced penetration rate of insecticides in the insect's epidermis, enhanced detoxification and metabolism of insects, and reduced sensitivity of target sites.

[0004] The insect cytochrome P450 enzyme system (P450 enzyme for short) belongs to the monooxygenase class and can participate in the metabolism and detoxification of exogenous substances (such as pesticides, plant secondary metabolites, etc.), and also participate in the synthesis and metabolism of insect endogenous substances. Piperonyl butoxide (piperonyl butoxide) is the most widely used P450 enzyme inhibitor, which has been on the market for many years as a pesticide synergist. Piperonyl butoxide is a semi-natural compound synthesized from safrole by simple synthesis. It can inhibit a variety of P450 enzymes related to biosynthesis reactions, thereby improving the insecticidal activity of a variety of pesticides, but has no obvious effect on P450 enzymes related to detoxification metabolism. Many studies have shown that the main reason for most pests to develop resistance and cross-resistance to pesticides is the enhancement of the action of P450 enzymes related to detoxification metabolism. Therefore, the development of P450 enzyme inhibitors related to detoxification metabolism is expected to play a synergistic role in pest control as a pesticide synergist. Summary of the invention

[0005] In order to enrich the types of existing pesticide synergists and further enhance the pest control effect, the present invention takes the CYP6AE48 enzyme of Spodoptera litura (a P450 enzyme related to detoxification metabolism) as the main target, and uses a method combining computer-aided drug design technology screening with biological effect screening to screen an amide compound, and proves that the compound has an enhanced or promoting effect on the control effect of high-efficiency chlorfenapyr and chlorantraniliprole. In order to achieve this technical purpose, the present invention adopts the following technical scheme.

[0006] Firstly, the present invention provides a use of an amide compound as a pesticide synergist, wherein the amide compound is N-{5-[2-(2-{3-[2-(2-methylphenoxy)ethoxy]benzyl}hydrazine)-2-oxoethyl]-1,3,4-thiadiazol-2-yl}benzamide.

[0007] Furthermore, in the use, the amide compound inhibits the cytochrome p450 enzyme of Spodoptera litura.

[0008] Furthermore, in the use, the Spodoptera litura cytochrome p450 enzyme is CYP6AE48 enzyme.

[0009] Based on the above use, another aspect of the present invention provides a method for improving the chemical control effect of Spodoptera litura, the method comprising: using the above amide compounds as Spodoptera litura cytochrome p450 enzyme inhibitors to act on Spodoptera litura.

[0010] Furthermore, in the method, the Spodoptera litura cytochrome p450 enzyme is CYP6AE48 enzyme.

[0011] Furthermore, in the method, the amide compound is dissolved in dimethyl sulfoxide to prepare a solution with a concentration of 10 g / L, and the solution is used in an amount of 20 μg / head.

[0012] Furthermore, the chemical control includes: using high-efficiency cyhalothrin for control and using chlorantraniliprole for control.

[0013] More specifically, at 24, 48, and 72 hours, the amide compounds increased the effect of cyhalothrin in controlling Spodoptera litura by 1.9343, 2.4639, and 2.2816 times; at 24 hours and 72 hours, the amide compounds increased the effect of chlorfenapyr in controlling Spodoptera litura by 1.6438 and 1.0128 times.

[0014] Compared with the prior art, the invention "use of an amide compound as a pesticide synergist" has the following beneficial effects:

[0015] The invention takes the CYP6AE48 enzyme of Spodoptera litura as the main target, adopts a method combining computer-aided drug design technology screening with biological effect screening, and screens out an amide compound, which is N-{5-[2-(2-{3-[2-(2-methylphenoxy)ethoxy]benzyl}hydrazine)-2-oxoethyl]-1,3,4-thiadiazol-2-yl}benzamide. Specifically, at 24, 48, and 72 hours, the compound improved the effect of cypermethrin on controlling Spodoptera litura by 1.9343, 2.4639, and 2.2816 times; at 24 hours and 72 hours, the amide compound improved the effect of chlorantraniliprole on controlling Spodoptera litura by 1.6438 and 1.0128 times, indicating that the compound can effectively inhibit the activity of the CYP6AE48 enzyme of Spodoptera litura, making it unable to normally perform the detoxification function of cypermethrin and chlorantraniliprole, thereby improving the control effect of cypermethrin and chlorantraniliprole. The compound can be used as a pesticide synergist to play a synergistic role in pest control.

[0016] On the other hand, existing P450 enzyme inhibitors (such as piperonyl butoxide) mainly inhibit P450 enzymes related to biosynthesis reactions, while P450 enzymes related to detoxification metabolism are the main reasons for most pests to produce resistance and cross-resistance to pesticides, and are more important for pest control. The present invention uses the P450 enzyme CYP6AE48 related to the detoxification metabolism of Spodoptera litura as a target, screens out a P450 enzyme inhibitor related to detoxification metabolism, and makes up for the technical defect of the relative lack of such enzyme inhibitors, which has important inspiration and guidance for the future research of pesticide synergists. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 is the chemical structural formula of the amide compound. DETAILED DESCRIPTION

[0018] The present invention is explained below in conjunction with the embodiments, and the technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0019] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0020] In the following examples, the amide compound is: N-{5-[2-(2-{3-[2-(2-methylphenoxy)ethoxy]benzyl}hydrazine)-2-oxoethyl]-1,3,4-thiadiazol-2-yl}benzamide (CAS No.: 380874-27-1, purity 99%), which was purchased from specs. The chemical structure of the compound is shown in Figure 1 .

[0021] Example 1

[0022] This example describes the killing effect of high-efficiency cyhalothrin alone on Spodoptera litura.

[0023] Test method: Weigh the high-efficiency cyhalothrin technical drug, prepare it into a 1 g / L stock solution with acetone as solvent, and then dilute it to 3, 2, 1.5, 1, and 0.5 mg / L respectively to obtain high-efficiency cyhalothrin solution.

[0024] The third instar larvae of Spodoptera litura were used as test insects, and the highly effective chlorfenapyr solution was dripped onto the pronotum of the larvae using a microdropper, with 1 μL dripped onto each larva. After natural air drying, the larvae were placed in an incubator with a temperature of 27±1°C, a relative humidity of 70%, and a light duration of 12 h. Acetone was used as the control group, and the rest of the treatment steps were the same. Twenty larvae were treated in each culture dish, repeated 3 times, and the results were observed after 24, 48, and 72 hours, and the number of dead insects was recorded. Larvae that did not wriggle when lightly touched with a brush were considered dead, and the toxicity regression equation, correlation coefficient, and LC were calculated using SPSS software. 50 .

[0025] Test results: See Table 1. As shown in Table 1, at 24, 48, and 72 hours, when high-efficiency cyhalothrin was used alone, LC 50 The values ​​are 2.5695, 2.3338 and 2.0676 mg / L respectively.

[0026] Table 1. Effects of using high-efficiency cyhalothrin alone

[0027] time Toxicity equation Correlation coefficient r (95% confidence interval) <![CDATA[LC 50 (mg / L)]]> 24h Y=2.6323X+3.9211 0.9944 2.5695 48h Y=2.5278X+4.0696 0.9838 2.3338 72h Y=2.8459X+4.1022 0.9921 2.0676

[0028] Example 2

[0029] This example describes the effect of using the amide compound when using high-efficiency cyhalothrin.

[0030] Test method: Weigh the high-efficiency cyhalothrin technical drug, use acetone as solvent to prepare a 1 g / L mother solution, and then dilute to 0.5, 1, 1.5, 2, 3 mg / L to obtain a high-efficiency cyhalothrin solution. Use dimethyl sulfoxide as solvent to prepare the amide compound at a concentration of 10 g / L.

[0031] The third instar larvae of Spodoptera litura were used as test insects, and the solution of the amide compound was dripped onto the pronotum of the larvae with a microdropper, with 20 μg dripped on each larva. After natural air drying for 1 hour, the highly effective chlorfenapyr solution was dripped in the same way, with 1 μl dripped on each larva. After the dripping, the insects were placed in an incubator with a temperature of 27±1°C, a relative humidity of 70%, and a light duration of 12 hours. The control group used dimethyl sulfoxide instead of the solution of the amide compound, and the other treatment steps were the same. Twenty insects were treated in each culture dish, repeated 3 times, and the results were observed after 24, 48, and 72 hours, and the number of dead insects was recorded. Larvae that did not wriggle when touched lightly with a brush were considered dead. SPSS software was used to calculate the toxicity regression equation, correlation coefficient, and LC 50 , and calculate the efficiency ratio.

[0032] Synergistic ratio (SR) = LC when using cyhalothrin alone 50 / LC of cyhalothrin when the amide compounds are added 50 .

[0033] Test results: See Table 2.

[0034] Table 2. The effect of using high-efficiency cyhalothrin when adding the amide compounds

[0035]

[0036]

[0037] It can be seen from Tables 1 and 2 that at 24, 48, and 72 hours, the LC values ​​of cyhalothrin when the amide compounds were added were 50 The values ​​were significantly lower than the LC when high-efficiency cyhalothrin was used alone. 50 , indicating that the amide compounds inhibit the CYP6AE48 enzyme of Spodoptera litura, making it unable to properly perform the detoxification function of high-efficiency cyhalothrin. Therefore, when adding the amide compounds, the effect of using high-efficiency cyhalothrin is better, which is in line with expectations. In addition, at 24, 48, and 72 hours, the synergistic multiples were 1.9343, 2.4639, and 2.2816 times, respectively, all greater than 1, which indicates that the use of the amide compounds has a good effect.

[0038] Example 3

[0039] This example describes the killing effect of chlorantraniliprole alone on Spodoptera litura.

[0040] Test method: Weigh the chlorantraniliprole technical drug, prepare it into a 1 g / L stock solution with acetone as solvent, and then dilute it to 1.2, 1, 0.8, 0.6, and 0.4 mg / L respectively to obtain chlorantraniliprole solution.

[0041] The third instar larvae of Spodoptera litura were used as the test insects. The chlorantraniliprole solution was dripped onto the pronotum of the larvae using a microdropper, with 1 μL dripped onto each larva. After natural air drying, the larvae were placed in an incubator with a temperature of 27±1°C, a relative humidity of 70%, and a light duration of 12 h. Acetone was used as the control group, and the rest of the treatment steps were the same. Twenty larvae were treated in each culture dish, repeated three times, and the results were observed after 24, 48, and 72 hours, and the number of dead insects was recorded. Larvae that did not wriggle when touched lightly with a brush were considered dead, and the toxicity regression equation, correlation coefficient, and LC were calculated using SPSS software. 50 .

[0042] Test results: See Table 3. As shown in Table 3, at 24, 48, and 72 hours, when chlorantraniliprole was used alone, LC 50 The values ​​are 1.6794, 0.5241 and 0.4721 mg / L respectively.

[0043] Table 3. Effect of using chlorantraniliprole alone

[0044]

[0045] Example 4

[0046] This example describes the effect of using the amide compounds when using chlorantraniliprole.

[0047] Test method: Weigh the chlorantraniliprole stock drug, use acetone as solvent to prepare a 1 g / L mother solution, then dilute to 1.2, 1, 0.8, 0.6, 0.4 mg / L to obtain a chlorantraniliprole solution. Use dimethyl sulfoxide as solvent to prepare the amide compound at a concentration of 10 g / L.

[0048] The third instar larvae of Spodoptera litura were used as test insects, and the solution of the amide compound was dripped onto the pronotum of the larvae with a microdropper, with 20 μg dripped on each larva. After natural air drying for 1 hour, the chlorantraniliprole solution was dripped in the same way, with 1 μl dripped on each larva. After the dripping, the insects were placed in an incubator with a temperature of 27±1°C, a relative humidity of 70%, and a light duration of 12 hours. The control group used dimethyl sulfoxide instead of the solution of the amide compound, and the other treatment steps were the same. Twenty insects were treated in each culture dish and repeated 3 times. The results were observed after 24, 48, and 72 hours, and the number of dead insects was recorded. Larvae that did not wriggle when touched lightly with a brush were considered dead. SPSS software was used to calculate the toxicity regression equation, correlation coefficient, and LC 50 , and calculate the efficiency ratio.

[0049] Synergistic ratio (SR) = LC when chlorantraniliprole is used alone 50 / LC of chlorantraniliprole when the amide compound is added 50 .

[0050] Test results: See Table 4.

[0051] Table 4. Effect of using chlorantraniliprole when adding the amide compounds

[0052]

[0053] It can be seen from Tables 3 and 4 that at 24h and 72h, the LC values ​​of chlorantraniliprole when the amide compounds are added are 50 The values ​​were lower than the LC when chlorantraniliprole was used alone. 50 Value, LC at 48h 50 The value is slightly higher, which may be due to experimental errors. The synergistic multiples at 24h and 72h were 1.6438 and 1.0128 times, respectively, indicating that the addition of the amide compounds has a certain improvement effect on the control effect of chlorantraniliprole.

[0054] In summary, the present invention takes the CYP6AE48 enzyme of Spodoptera litura as the main target, and adopts a method combining computer-aided drug design technology screening with biological effect screening to screen out an amide compound, which is N-{5-[2-(2-{3-[2-(2-methylphenoxy)ethoxy]benzyl}hydrazine)-2-oxoethyl]-1,3,4-thiadiazol-2-yl}benzamide. It is known from experiments that the compound has a good inhibitory effect on the CYP6AE48 enzyme of Spodoptera litura, preventing it from degrading highly effective chlorfenapyr and chlorantraniliprole, and is expected to play a synergistic role in pest control as a pesticide synergist.

[0055] The embodiments described above are only some embodiments of the present invention, not all embodiments. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but only represents selected embodiments of the present invention. All other embodiments obtained without creative work and related deductions and substitutions made by ordinary technicians in the field under the conditions of the concept of the present invention belong to the scope of protection of the present invention.

Claims

1. A use of an amide compound as a pesticide synergist, characterized in that: The amide compound is N-{5-[2-(2-{3-[2-(2-methylphenoxy)ethoxy]benzyl}hydrazine)-2-oxoethyl]-1,3,4-thiadiazol-2-yl}benzamide.

2. The use according to claim 1, characterized in that The amide compounds inhibit Spodoptera litura cytochrome p450 enzymes.

3. The use according to claim 2, characterized in that: The Spodoptera litura cytochrome p450 enzyme is CYP6AE48 enzyme.

4. A method for improving the chemical control effect of Spodoptera litura, characterized in that: The method comprises: using an amide compound as a cytochrome p450 enzyme inhibitor of Spodoptera litura to act on Spodoptera litura; The amide compound is N-{5-[2-(2-{3-[2-(2-methylphenoxy)ethoxy]benzyl}hydrazine)-2-oxoethyl]-1,3,4-thiadiazol-2-yl}benzamide.

5. The method according to claim 4, characterized in that The Spodoptera litura cytochrome p450 enzyme is CYP6AE48 enzyme.

6. The method according to claim 4, characterized in that The amide compound was dissolved in dimethyl sulfoxide to prepare a solution with a concentration of 10 g / L. The amount of the solution used was 20 μg / head.

7. The method according to claim 4, characterized in that The chemical control includes: using highly effective cyhalothrin for control and using chlorantraniliprole for control.

8. The method according to claim 7, characterized in that At 24, 48 and 72 hours, the amide compounds increased the effect of cyhalothrin on controlling Spodoptera litura by 1.9343, 2.4639 and 2.2816 times.

9. The method according to claim 7, characterized in that: At 24h and 72h, the amide compounds increased the effect of chlorantraniliprole in controlling Spodoptera litura by 1.6438 and 1.0128 times.

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