Diester group substituted methyl salicylate analogue, and preparation method and application thereof
阅读说明：本技术 一类双酯基取代的水杨酸甲酯类似物及其制备方法和应用 (Diester group substituted methyl salicylate analogue, and preparation method and application thereof ) 是由 杨新玲 杨朝凯 潘世香 刘彦 秦耀果 凌云 石卓 于 2021-10-29 设计创作，主要内容包括：本发明提供一类双酯基取代的水杨酸甲酯类似物及其制备方法和应用。其结构式如式A所示。本发明的双酯基取代的水杨酸甲酯类似物,制备方法简便易行,操作简单,产物纯化容易,成本较低,具有多重的生物活性。不仅对蚜虫具有作用,还有吸引蚜虫天敌的潜力。可以作为一类蚜虫及其天敌的行为控制剂,应用于农业害虫的绿色防控。(The invention provides diester-group substituted methyl salicylate analogues and a preparation method and application thereof. The structural formula is shown as formula A. The diester-substituted methyl salicylate analogue has simple and easy preparation method and simple operationThe product is easy to purify, the cost is low, and multiple biological activities are achieved. Not only has the function on aphids, but also has the potential of attracting natural enemies of the aphids. Can be used as a behavior control agent of aphids and natural enemies thereof and applied to green prevention and control of agricultural pests.)
1. A compound of formula A:
in the formula A, R1Is C1-C12Straight or branched alkyl, halo C1-C12Straight or branched alkyl, C1-C12Alkoxy, halo C1-C12Alkoxy radical, C3-C12Any one of cycloalkyl and aromatic heterocycle;
R2is a mono-or poly-substituted,
R2independently selected from H, OH, C1-C12Straight or branched alkyl, halo C1-C12Alkyl radical, C1-C12Alkoxy, halo C1-C12Any one of the group consisting of alkoxy, halogen and nitro;
x is O, S.
2. The compound of claim 1, wherein: the diester-substituted methyl salicylate analog shown in the formula A is any one of the following substances:
。
3. a process for preparing a compound according to claim 1 or 2, comprising the steps of:
in the presence of an acid binding agent, carrying out a condensation reaction on substituted acyl chloride shown in a formula C and a compound shown in a formula B in an organic solvent to obtain a compound shown in a formula A:
in the formula B, R2X and R of formula A2And X are the same;
r of the formula C1With R of the formula A1The same;
or the like, or, alternatively,
carrying out esterification condensation reaction on a substituted acid shown as a formula D and a compound shown as a formula B in an organic solvent in the presence of a dehydrating agent to obtain a compound shown as a formula A:
in the formula B, R2X and R of formula A2And X are the same;
r of the formula D1With R of the formula A1The same is true.
4. The method of claim 3, wherein: the molar ratio of the substituted acyl chloride to the compound shown in the formula B and the acid-binding agent is 1:1-2:1-2 in sequence;
the molar ratio of the substituted acid to the compound shown in the formula B and the dehydrating agent is 1:1:1-3 in sequence.
5. The method according to claim 3 or 4, characterized in that: the reaction temperature is-20 ℃ to 100 ℃ and the reaction time is 0.5 to 5 hours;
the acid-binding agent is selected from at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, cesium carbonate, potassium carbonate, triethylamine, N' -diisopropylethylamine and pyridine;
the dehydrating agent is one or a mixture of more of dicyclohexylcarbodiimide, N-diisopropylcarbodiimide and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide.
6. The use of a compound according to claim 1 or 2 for the control of pests on crops, fruit trees, Chinese medicinal herbs and flowers, said pests being aphids.
7. The use of a compound according to claim 1 or 2 for the attraction of natural enemies of pests of crops, fruit trees, Chinese herbal medicines and flowers, said natural enemies of pests being natural enemies of aphids, and said natural enemies of pests being in particular ladybug and chrysopidae.
8. A medicament containing a compound according to claim 1 or 2.
9. The medicament of claim 8, wherein: the drug is an aphid control agent, a ladybug control agent or a chrysopa perla control agent.
10. The medicament of claim 9, wherein: the aphid control agent is an aphid repellent; the ladybug control agent is a ladybug attractant; the chrysopa perla control agent is a chrysopa perla attractant.
Technical Field
The invention belongs to the field of agricultural chemicals, and particularly relates to diester-substituted methyl salicylate analogues, a preparation method thereof, and application of the compounds in aphid control and natural enemy attraction.
Background
Aphids are one of the major pests in agriculture and cause very serious damage to plants. Aphids are harmful to plants mainly by sucking plant juice, and more seriously, aphids can also spread various plant viruses, thereby bringing huge economic loss to agricultural production. Therefore, the effective control of aphids can achieve the purposes of controlling pests and preventing diseases, and has important significance in agricultural production. The prevention and control of aphids is mainly carried out by chemical pesticide control methods, and the dosage of the aphids is increased due to the resistance of the aphids to traditional chemical aphicides, so that the toxicity of some chemical aphicides to non-target organisms such as bees causes a great risk to the environmental safety and ecological safety of the traditional chemical aphicides (Woodcock, B. A. science,2017,356, 1393-. Therefore, on the premise of being eco-friendly, the development of a high-efficiency aphid control agent with a novel action mechanism and a novel structure is very important.
When the plants are damaged by pests, a chemical signal molecule is released to antagonize the feeding pests and attract the arrival of natural enemies of the pests. This class of chemical signaling molecules is called "inducible plant volatiles" (Ted C.J.Turlings et al. Annu. Rev. Entomol.2018,63, 433-452). More than 2000 volatile components of over 90 plants have been identified, many of which are produced and secreted by plants for self defense, and plant protection is carried out by using the characteristics of plant volatiles, and has the advantages of trace amount, high selectivity, ecological safety and the like (Yavanna Aartsma et al. New Phytol.2017,216, 1054-1063).
Various plant volatiles have been reported to have antagonistic effects on aphids, including those of broad bean (Vicia faba), Ocimum basilicum (Ocimum basilicum), Capsicum annuum (Capsicum annuum) and Hemizygia pepatia (Ashby)) plants (Webster B. anim Behav,2010,79,451 457; Dardouri T. Pest Manag Sci.2019,75:1571 1584; Bruce T. Pest Manag Sci,2005,61, 1115-. Among them, methyl salicylate (formula I) is a classical plant volatile substance antagonistic to aphids, and Hardie et al reported that Vicia faba can release methyl salicylate to produce a repellent effect on Aphis niger (Hardie J.J Chem Ecol, 1994,20, 2847-2855.).
Rachel E.Mallinger reported that methyl salicylate could attract the natural enemy chrysopa perla (Mallinger R E.journal of Econ Entomol,2011, 115-124.). Junwei Zhu reported that methyl salicylate can attract the natural aphid coccinella septempunctata (Zhu J.J Chem Ecol,2005,31, 1733-. In addition, s.e. halobert et al reported that various natural alcohols including geraniol (formula II) and farnesol (formula III) have repellent effects on aphids (halobert, s.e. ann Appl biol.2009,154, 303-307.). However, natural plant volatiles have limited sources, unstable structure and short shelf life, and thus have limitations in practical application. Therefore, the improvement research on the structure of plant natural volatile matters is carried out, and the aphid control agent with better shelf life and application is found to be of great significance.
Disclosure of Invention
It is an object of the present invention to provide a class of diester-substituted methyl salicylate analogs.
The structural formula of the diester-substituted methyl salicylate analogue provided by the invention is shown as the formula A:
in the formula A, the reaction solution is prepared,R1can be C1-C12Straight or linear alkyl (specifically may be C)1-C6Straight or straight chain alkyl, e.g. -CH3、 -CH2CH3、-(CH2)2CH3、-(CH2)3CH3、-(CH2)4CH3、-(CH2)5CH3、-CH(CH3)2) Halogen substituted C1-C12Straight or straight chain alkyl, C1-C12Alkoxy, halo C1-C12Alkoxy radical, C3-C12Cycloalkyl (which may be C in particular)3-C6Cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl), aromatic heterocycle (such as thiophene);
R2is a mono-or poly-substituted,
R2independently selected from H, OH, C1-C12Straight or linear alkyl (specifically may be C)1-C6Straight or branched alkyl), halo C1-C12Alkyl (e.g. trifluoromethyl), C1-C12Alkoxy (which may be C in particular)1-C6Alkoxy, e.g. methoxy), halo C1-C12Any one of the group consisting of alkoxy, halogen and nitro;
x may be O, S, specifically O.
The diester-substituted methyl salicylate analog represented by the formula A can be any one of the following substances:
the diester-substituted methyl salicylate analog shown in the formula A is prepared by a method comprising the following steps:
in the presence of an acid binding agent, carrying out a condensation reaction on substituted acyl chloride (shown as a formula C) and a compound shown as a formula B in an organic solvent to obtain a compound shown as a formula A:
in the formula B, R2X and R of formula A2And X are the same;
r of the formula C1With R of the formula A1The same;
the diester-substituted methyl salicylate analog of formula a can also be obtained by a preparation method comprising the steps of:
carrying out esterification condensation reaction on a substituted acid (formula D) and a compound shown as a formula B in an organic solvent in the presence of a dehydrating agent to obtain a compound shown as a formula A:
in the formula B, R2X and R of formula A2And X are the same;
r of the formula D1With R of the formula A1The same;
in the method, the molar ratio of the compound shown in the formula B to the substituted acyl chloride and the acid-binding agent can be 1:1-2:1-2 in sequence; specifically 1:1: 1; 1:1: 1.5; 1:1: 2; 1:1.5: 2; 1:2: 2;
the molar ratio of the compound shown in the formula B to the substituted acid and the dehydrating agent can be 1:1:1-3 in sequence, and specifically can be 1:1: 1; 1:1: 1.2; 1:1: 1.5; 1:1: 2; 1:1: 3;
the reaction temperature can be-20-100 ℃, and specifically can be 0-40 ℃; the time is 0.5-5 h;
the acid-binding agent can be at least one of sodium hydroxide, potassium hydroxide, sodium carbonate, cesium carbonate, potassium carbonate, triethylamine, N' -diisopropylethylamine and pyridine;
the dehydrating agent can be one or a mixture of more of dicyclohexylcarbodiimide, N-diisopropylcarbodiimide and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide;
the organic solvent can be at least one of dichloromethane, chloroform, carbon tetrachloride, N-dimethylformamide, benzene, toluene, xylene, tetrahydrofuran, pyridine, acetonitrile, methanol, ethanol, N-propanol, isopropanol, methyl acetate, ethyl acetate, diethyl ether and diisopropyl ether.
The invention also aims to provide the application of the diester-group-substituted methyl salicylate analogue in the prevention and control of pests such as crops, fruit trees, Chinese herbal medicines and flowers, wherein the pests are aphids.
The invention also aims to provide the application of the diester-group-substituted methyl salicylate analogue in attracting natural enemies of crops, fruit trees, Chinese herbal medicines and flowers, wherein the natural enemies of the pests are natural enemies of aphids, and the natural enemies of the pests can be ladybug and chrysopidae.
The invention also provides a medicament which contains the diester-substituted methyl salicylate analogue.
The drug can be an aphid control agent, a ladybug control agent or a chrysopa perla control agent.
The aphid control agent can be an aphid repellent.
The ladybug control agent can be a ladybug attractant.
The chrysopa perla control agent can be a chrysopa perla attractant.
The diester-substituted methyl salicylate analogue has the advantages of simple and easy preparation method, simple operation, easy product purification, lower cost and multiple biological activities. Not only has the function on aphids, but also has the potential of attracting the natural enemies of the aphids.
According to the invention, an active substructure splicing method is adopted, methyl salicylate (I) and geraniol (II) which are plant volatile substances are spliced, geraniol is used for replacing methyl in a methyl salicylate structure to obtain geranyl salicylate, and then phenolic hydroxyl groups on salicylic acid fragments are further esterified and transformed, so that a diester-based methyl salicylate analogue (formula A) is finally discovered. The compound of the formula A has the advantages of simple and easy preparation method, better activity, multiple functions and enhanced stability, can be used as a behavior control agent of aphids and natural enemies thereof, and is applied to green prevention and control of agricultural pests.
Detailed Description
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The present invention provides diester-substituted methyl salicylate analogs of formula a:
in the formula A, R1Can be C1-C12Straight or linear alkyl (specifically may be C)1-C6Straight or straight chain alkyl, e.g. -CH3、-CH2CH3、-(CH2)2CH3、-(CH2)4CH3、-(CH2)5CH3、-CH(CH3)2) Halogen substituted C1-C12Straight or straight chain alkyl, C1-C12Alkoxy, halo C1-C12Alkoxy radical, C3-C12Cycloalkyl (which may be C in particular)3-C6Cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl), aromatic heterocycle (such as thiophene, pyrazole);
R2is a mono-or poly-substituted,
R2independently selected from H, OH, C1-C12Straight or linear alkyl (specifically may be C)1-C6Straight or branched alkyl), halo C1-C12Alkyl (e.g. trifluoromethyl), C1-C12Alkoxy (which may be C in particular)1-C6An alkoxy group,such as methoxy), halogeno C1-C12Any one of the group consisting of alkoxy, halogen and nitro;
x may be O, S, specifically O.
The invention also provides application of the diester-group-substituted methyl salicylate analogue shown in the formula A in prevention and control of pests such as crops, fruit trees, Chinese herbal medicines and flowers, wherein the pests are aphids.
The invention also provides application of the diester-group-substituted methyl salicylate analogue shown in the formula A in attracting natural enemies of pests of crops, fruit trees, Chinese herbal medicines and flowers, wherein the natural enemies of the pests are natural enemies of aphids, and the natural enemies of the pests can be ladybug and chrysopa perla.
The invention also provides a medicament which contains the diester-substituted methyl salicylate analogue.
The drug can be an aphid repellent, a ladybug attractant or a chrysopa perla attractant.
The diester-substituted methyl salicylate analogue has the advantages of simple and easy preparation method, simple operation, easy product purification, lower cost and multiple biological activities. Not only has the function on aphids, but also has the potential of attracting the natural enemies of the aphids.
Example 1 preparation of 1- (E) -3, 7-dimethyl-2, 6-octadiene-2- (acetoxy) -3-methoxybenzoate (A-01)
In a 100mL three-necked flask, 2.18g of 3-methoxysalicylic acid, 3.22g of DCC (dicyclohexylcarbodiimide), and 1.74g of DMAP (4-dimethylaminopyridine) were added. The resulting mixture was dissolved in 40mL of tetrahydrofuran, and 2.00g of geraniol was added dropwise thereto, followed by reaction at 20 ℃ for 3 hours. Extracting with water for three times to obtain organic phase, removing solvent under reduced pressure, and separating with silica gel column chromatography (the eluent is petroleum ether and ethyl acetate ═ V)1：V250:1) to give 1- (E) -3, 7-dimethyl-2, 6-octadiene-2-hydroxy-3-methoxybenzoate as a yellow liquid in 53.8% yield.
In a 50mL three-necked flask, 2g of 1- (E) -3, 7-dimethyl-2, 6-octadiene-2-hydroxy-3-methoxybenzoate (6.6m mol) was added, and then 30mL of methylene chloride was added to dissolve and stir well. Then, 1.34g of triethylamine (13.2m mol) is added as an acid-binding agent, and the temperature is reduced in an ice bath. 1.05g of acetyl chloride (13.2m mol) is dissolved in 20mL of anhydrous dichloromethane, after uniform mixing, the mixture is slowly dripped into a three-necked flask, ice bath is removed, and the mixture is reacted at 40 ℃ for 3 hours. After the reaction, the reaction mixture was washed with water 3 times, extracted with dichloromethane, and dried over anhydrous sodium sulfate. And (4) carrying out column chromatography separation to obtain the target compound A-01. A-01 was a colorless liquid, yield 76.56%.
Example 2 preparation of 1- (E) -3, 7-dimethyl-2, 6-octadiene-2- (propionyloxy) -3-methoxybenzoate (A-02)
In a 100mL three-necked flask, 2.18g of 3-methoxysalicylic acid, 3.22g of DCC (dicyclohexylcarbodiimide), and 1.74g of DMAP (4-dimethylaminopyridine) were added. The resulting mixture was dissolved in 40mL of tetrahydrofuran, and 2.00g of geraniol was added dropwise thereto, followed by reaction at 20 ℃ for 3 hours. Extracting with water for three times to obtain organic phase, removing solvent under reduced pressure, and separating with silica gel column chromatography (the eluent is petroleum ether and ethyl acetate ═ V)1：V250:1) to give 1- (E) -3, 7-dimethyl-2, 6-octadiene-2-hydroxy-3-methoxybenzoate as a yellow liquid in 53.8% yield.
In a 50mL three-necked flask, 2g of 1- (E) -3, 7-dimethyl-2, 6-octadiene-2-hydroxy-3-methoxybenzoate (6.6m mol) was added, and then 30mL of tetrahydrofuran was added to dissolve and stir well. Then 0.782g pyridine (9.9m mol) is added as an acid-binding agent, and the temperature is reduced in an ice bath. 1.22g of propionyl chloride (13.2m mol) is dissolved in 20mL of anhydrous tetrahydrofuran, after uniform mixing, the mixture is slowly dripped into a three-necked bottle, ice bath is removed, and reaction is carried out for 3h at the normal temperature of 30 ℃. After the reaction, the reaction mixture was washed with water 3 times, extracted with dichloromethane, and dried over anhydrous sodium sulfate. And (4) carrying out column chromatography separation to obtain the target compound A-01. A-02 was a colorless liquid, yield 73.97%.
Example 3 preparation of 1- (E) -3, 7-dimethyl-2, 6-octadiene-2- (butyryloxy) -3-methoxybenzoate (A-03)
In a 100mL three-necked flask, 2.18g of 3-methoxysalicylic acid, 3.22g of DCC (dicyclohexylcarbodiimide), and 1.74g of DMAP (4-dimethylaminopyridine) were added. The resulting mixture was dissolved in 40mL of tetrahydrofuran, and 2.00g of geraniol was added dropwise thereto, followed by reaction at 20 ℃ for 3 hours. The organic phase was extracted three times with water, and the organic phase was removed under reduced pressure, and separated by silica gel column chromatography (eluent: petroleum ether: ethyl acetate: V1: V2: 50:1) to give 1- (E) -3, 7-dimethyl-2, 6-octadiene-2-hydroxy-3-methoxybenzoate, yield 53.8% as a yellow liquid.
In a 50mL three-necked flask, 2g of 1- (E) -3, 7-dimethyl-2, 6-octadiene-2-hydroxy-3-methoxybenzoate (6.6m mol) was added, and then 30mL of ethyl acetate was added to dissolve and stir well. Then, 0.58g of butyric acid (6.6m mol) and 30ml of an ethyl acetate solution were added to dissolve uniformly, stirred, and ice-cooled to 0 ℃. 0.83g N, N-diisopropylcarbodiimide (6.6m mol) and 0.08g of 4-dimethylaminopyridine were weighed into a 50ml beaker and dissolved by adding 20ml of a chloroform solution. Then, the liquid in the beaker is added into the three-mouth reaction bottle dropwise, the ice bath is removed, and the reaction is carried out for 8 hours at the temperature of 30 ℃. After the reaction is finished, filtering to remove white solid matters in the reaction bottle, concentrating the filtrate, and performing column chromatography to obtain A-03. A-03 was a colorless liquid, yield 60.18%.
Example 4 preparation of 1- (E) -3, 7-dimethyl-2, 6-octadiene-2- (hexanoyloxy) benzoate (A-17)
In a 100mL three-necked bottle, 1.79g of salicylic acid, 3.22g of DCC (dicyclohexylcarbodiimide), and 1.74g of DMAP (4-dimethylaminopyridine) were added. The mixture was dissolved in 40mL of tetrahydrofuran, and 2.00g of geraniol was added dropwise and reacted at 20 ℃ for 3 hours. The organic phase was extracted three times with water, and the organic phase was removed under reduced pressure, and separated by column chromatography on silica gel (eluent: petroleum ether: ethyl acetate: V1: V2: 50:1) to give 1- (E) -3, 7-dimethyl-2, 6-octadiene-2-hydroxy-3-methoxybenzoate, yield 63.15% as a yellow liquid.
In a 50mL three-necked flask, 1.81(E) -3, 7-dimethyl-2, 6-octadiene-2-hydroxybenzoate (6.6m mol) was added, and then 30mL of dichloromethane was added to dissolve and stir well. Then 0.77g hexanoic acid (6.6m mol) and 30ml dichloromethane solution were added to dissolve homogeneously, stirred and ice-cooled to 0 ℃. 2.72 g of dicyclohexylcarbodiimide (13.2ml) and 0.08g of 4-dimethylaminopyridine were weighed into a 50ml beaker and dissolved by adding 20ml of a dichloromethane solution. Then, the liquid in the beaker is added dropwise into the three-mouth reaction bottle, the ice bath is removed, and the reaction lasts for 8 hours at 40 ℃. After the reaction is finished, filtering to remove white solid matters in the reaction bottle, concentrating the filtrate, and carrying out column chromatography to obtain A-17. A-13 was a colorless liquid, yield 72.57%.
A-04 to A-16 and A-18 can be prepared by the method for preparing A-01, A-02, A-03 or A-17 in the above embodiment.
The structure and the basic physicochemical data of the compound of formula a are given in table 1 below, and the nmr hydrogen spectra data of the compound of formula a in table 2 below, all prepared according to a method similar to that described in the preceding examples 1, 2, 3 and 4.
TABLE 1 Structure, shape and yield of Compounds of formula A
TABLE 2 NMR Hydrogen Spectroscopy data (500MHz, CDCl) for Compounds of formula A3)
Example 5 repellent Activity of Compounds of formula A against aphids
In this example, A-01-A-12, A-17 and the like were used as representative compounds (but not limited to these compounds) to test the aphid repelling activity.
A T-tube olfactometer is used for testing (the testing method refers to Al, A.S.J.chem.Ecol.2000,26, 1765-1771.), more than 40 wings of the green peach aphids are released from a release port, each arm is pumped in humid air which passes through activated carbon and distilled water at 0.2L/min through an air pump and firstly passes through 5 mu g of sample flavor source substances (the compound shown in the formula A), the other arm is used as a contrast arm, and the introduced humid air firstly passes through a solvent. The number of aphids on each arm at 15min of sample introduction was recorded. The olfactometer and the skin tube are cleaned by absolute ethyl alcohol every time of repeating, the filter paper sheet is replaced and used for exchanging two arms, and each sample experiment is repeated for four times. The control group or the treatment group was counted by crossing the center of the olfactometer by 2cm, and aphids that did not cross were counted as non-reaction groups.
Calculating the corrected repelling rate, wherein the formula is shown as the following formula:
the corrected repellency rate is (number of insects in control group-number of insects in treatment group)/(number of insects in control group + number of insects in treatment group) × 100%
TABLE 3 repellent Activity of Compounds of formula A against Myzus persicae (5. mu.g)
The results in Table 3 show that the compound of the invention has repellent activity on peach aphids at a low dose of 5 mu g, wherein the repellent rate of the compounds A-04, A-05, A-11 and A-17 on the peach aphids is more than 70%, and the repellent activity of natural plant volatile methyl salicylate and geraniol on the peach aphids is 35.94 and 29.13%. The repellent activity of the compound of the invention to aphids is obviously improved compared with the natural lead substance.
Example 6 protein binding of Compounds of formula A to Harxy. OBP15 of harmonia axyridis
This example represents A-01-A-12 and A-17 (but not limited to these compounds) for protein binding activity assay with Harxy
The test material comprises 2 mu mol/L of water solution of harmonia axyridis OBP15 protein, fluorescent probe molecules: n-phenyl-1-naphthylamine (1-NPN probe molecule)
The testing device comprises: agilent fluorescence spectrophotometer
The test method comprises the following steps: will 1Preparing the NPN probe molecules and the compound to be detected into 1m mol/L solution to be detected by using chromatographic grade methanol. The fluorescence was measured using a pipette gun to add 2mL of aqueous solution of haxy. obp15 protein to the cuvette and 4 μ L of probe molecule. And 4, 8, 12, 16, 20, 24, 28, 32, 36, 40 μ L of test compound was added stepwise in subsequent tests. After each addition of test compound, the absorbance in the cuvette was again measured using a fluorescence spectrophotometer. The competitive binding capacity of the compound to the "protein-probe" was calculated by the decrease in fluorescence. And calculates its IC50The results are shown in Table 4.
TABLE 4 protein binding data of Compounds of formula A to harmonia axyridis OBP15
TABLE 4 IC of Compounds of formula A50The numerical value shows that the combination ability of the compound in the formula A and the harmonia axyridis Haxy. OBP15 is generally better than that of the natural attractant substance methyl salicylate of the harmonia axyridis, which indicates that the compound has potential behavior regulation ability on the harmonia axyridis.
Example 7 attractive Activity of part of the Compound of formula A against Harmonia axyridis at Low doses
In this example, only A-05 is taken as a representative compound (but not limited to the compound), and the insect behavior box is used for carrying out the test of the attractive activity of the ladybug (the test method is referred to Al, A.S.J.chem.Ecol.2000,26, 1765-1771.) so as to represent the attractive activity of the compound of the formula A to the harmonia axyridis at a low dose.
The insect behavior box is formed by connecting a release bottle, a treatment bottle and a blank control bottle 3. The treatment vial and the blank control vial were positioned on either side of the release vial. Dissolving the compound A-05 to be detected with paraffin oil, and preparing into a solution to be detected of 10 mg/mL. 10 mu L of solution to be detected is dripped on a filter paper strip with the thickness of 10mm x 30mm and is placed in a processing bottle. A filter paper strip dropwise added with paraffin oil is arranged in a blank control bottle. The treatment flask and the blank control flask were purged with air at 0.3L/min using a suction pump with humidified air containing activated carbon and distilled water. In the release bottle, 40 heads of female or male harmonia axyridis are released. The numbers of harmonia axyridis in the treated and blank control bottles were recorded at 30min, 60min, 90min and 120min of introduction.
After the experiment was completed, the whole apparatus was cleaned using soapy water and absolute ethyl alcohol, and each experiment was repeated 3 times. Harmonia axyridis that did not enter the treatment and blank control bottles was recorded as an unreacted group.
The selectivity of harmonia axyridis was calculated using the following formula:
the selection rate is 100 percent of the number of the ladybug in the treated bottle/(the number of the ladybug in the treated bottle + the number of the ladybug in the blank control bottle)
TABLE 5 attraction of Compounds of formula A (A-05) to Lawsonia axyridis
In this experiment, the literature reports butyl acrylate, an active substance that has an attractive effect on the Male harmonia axyridis, and methyl salicylate, a natural volatile substance, as positive controls. The compound A-05 with better binding activity to the harmonia axyridis protein is selected as a representative compound to be tested. The results in Table 5 show that the selectivity of the harmonia axyridis to A-05 is higher than that of methyl salicylate and butyl acrylate. Especially at 90min, the best effect is achieved, which shows that A-05 has attraction effect on the male harmonia axyridis.
TABLE 6 attraction of Compound A (A-05) of formula A to female harmonia axyridis
Analysis of the results in Table 6 shows that the A-05 selectivity of female harmonia axyridis is comparable to that of methyl salicylate, slightly lower than that of butyl acrylate, at 60 or 90 minutes. The result shows that the attractive activity of the A-05 compound to the female is closer to that of methyl salicylate and slightly lower than that of butyl acrylate.
The results in tables 5 and 6 show that the attracting activity of A-05 to male harmonia axyridis is superior to that to female harmonia axyridis.
Example 8 Low dose activation of Compounds of formula A on Anthragma chrysosporium
This example uses only A-05 and A-17 as representative compounds (but not limited to them) and the related experimental tests were carried out using an antennal electrophysiological instrument (test methods are described in Junwei, Zhu. J. chem. Ecol.1999, 25, 1163-1177.) to show the antennal activation of chrysopa pallens at low doses of the compound of formula A.
Cutting off the antenna of the chrysopa perla, and cutting off a tiny section at each of two ends of the antenna to expose the internal tissue of the antenna. And adhering the cut antenna on two sides of the antenna electrode by using conductive adhesive and connecting the antenna electrode with the antenna electrophysiological equipment. The compound to be tested is prepared into 100, 1000 and 10000 mu g/mL solution to be tested by using normal hexane.
During testing, 10 mu L of n-hexane is firstly dripped on the filter paper strip and blown to the antenna through the air blowing hole of the device. The response potential (mV) of the solvent to the antenna was recorded. Then, 10. mu.L of the test solution was dropped on a filter paper strip, blown against the antenna, and the response potential (mV) of the compound solution to the antenna was recorded.
The effect of the compound on the antenna was calculated using the following formula:
response value (mV) ═ antenna response value of compound solution-antenna response value of solvent
TABLE 7 low dose of compounds of formula A on chrysosporium antennapedia activation values
It can be found from the results of Table 7 that A-05 and A-17 can effectively activate the antennal electrophysiological response value of chrysopa perla, wherein A-05 is more obvious and is obviously higher than the natural substances of geraniol and methyl salicylate at dosage levels of 1 mug, 10 mug and 100 mug. The A-17 compound was also significantly higher than the two above natural substances at 1. mu.g and 10. mu.g levels. The compound of the formula A can effectively activate the antennal physiological potential of the chrysopa perla and has attraction effect on the chrysopa perla.
In conclusion, the compound of the formula A has a repelling effect on aphids, can be efficiently combined with OBP15 protein of harmonia axyridis, has an attraction effect on harmonia axyridis, can activate an antennal potential of chrysopa sinica and has an attraction effect on chrysopa sinica. Therefore, the compound of the formula A has double functions of repelling aphids and attracting natural enemies of the aphids, can provide a new strategy for preventing and controlling field aphids, and is favorable for realizing green prevention and control of the aphids.