A 2-cyanoacrylate compound, a preparation method thereof, and an application thereof in pesticides

Abstract

The present invention belongs to the field of pesticides and relates to a 2-cyanoacrylate compound, a preparation method thereof, and its application in pesticides. 4-Cyanophenol undergoes an alcoholysis reaction with an anhydrous ethanol solution of hydrogen chloride to obtain an intermediate of ethyl 4-hydroxybenzimidate hydrochloride; subsequently, the intermediate of ethyl 4-hydroxybenzimidate hydrochloride undergoes a condensation reaction with ethyl cyanoacetate under the action of a base to obtain a 2-cyanoacrylate compound I; the 2-cyanoacrylate compound I reacts to form a salt under the action of a base to obtain a derivative II of the 2-cyanoacrylate compound. The novel 2-cyanoacrylate compound I and the derivative II of the 2-cyanoacrylate compound provided by the present invention have good bactericidal activity, and have the advantages of broad-spectrum bactericidal and high efficiency; at the same time, they have low toxicity to non-target organisms, have good environmental compatibility, can be naturally and rapidly degraded in the environment, will not cause residual harm to the environment, and have high safety.

Description

Technical Field

[0001] The present invention belongs to the field of pesticides and relates to a 2-cyanoacrylate compound, a preparation method thereof, and its application in pesticides. Background Art

[0002] 2-Cyanoacrylate compounds have been widely used in the fields of medicine and pesticides due to their good biological activities. For example, the fungicide variety, phenamacril, can effectively control diseases such as wheat head blight and rice bakanae disease.

[0003] Patent document CN1160318C discloses a class of 2-cyano-3-substituted phenyl acrylate compounds represented by the following general formula (A), and this class of compounds has control effects on various diseases caused by Fusarium spp. such as wheat head blight.

[0004]

[0005] Patent document CN101381326A discloses a class of 2-cyano 3-(substituted) amino-3-phenyl acrylate compounds represented by the following general formula (B), and this compound has control effects on Fusarium spp. and rust fungi.

[0006]

[0007] Patent document CN109879834A discloses a class of 3-amino-2-cyanoacrylate compounds represented by the following general formula (C), and this class of Mannich base fungicides has good control effects on Fusarium spp.

[0008]

[0009] Patent document CN109879841B discloses a class of (Z)-3-imino-1-propenol compounds represented by the following general formula (D), and this class of compounds has control effects on wheat head blight causing plant diseases and can reduce the production of deoxynivalenol (DON). The compound of No. 8 structure in this patent is different from the compound I of the present patent.

[0010]

[0011] Patent document CN109867623A discloses a class of 3-pyridyl 3-amino-2-cyanoacrylate compounds represented by the following general formula (E), and this compound has control effects on wheat scab.

[0012]

[0013] Patent document CN 114790152A discloses a class of 2-cyanoacrylate compounds represented by the following general formula (F), and this compound has a control effect on Fusarium fungi.

[0014]

[0015] However, in the prior art, the bactericidal spectrum of cyenopyrafen is very narrow, and it has no control effect on other serious plant fungal diseases in agriculture such as rice blast, gray mold, anthracnose, etc., which seriously affects its large-scale application in agricultural disease control and seriously affects its economic value. Therefore, how to expand its antibacterial spectrum, improve its application value and reduce its toxicity to non-target organisms is the direction and hot spot of current pesticide development. SUMMARY OF THE INVENTION

[0016] The technical problem to be solved by the present invention is to provide, in view of the deficiencies of the prior art, a novel 2-cyanoacrylate compound and its derivatives with broad-spectrum bactericidal activity, relatively high bactericidal activity and relatively low toxicity to non-target organisms, for controlling fungal diseases of crops.

[0017] To solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0018] The present invention discloses a 2-cyanoacrylate compound, and the structural formula of the 2-cyanoacrylate compound is shown in Formula I:

[0019]

[0020] Further, the present invention discloses a preparation method of the above 2-cyanoacrylate compound. 4-Cyanophenol reacts with an anhydrous ethanol solution of hydrogen chloride to obtain an intermediate of ethyl 4-hydroxybenzimidate hydrochloride; subsequently, the intermediate of ethyl 4-hydroxybenzimidate hydrochloride undergoes a condensation reaction with ethyl cyanoacetate under the action of a base to obtain the 2-cyanoacrylate compound I;

[0021]

[0022] In some embodiments, the molar ratio of 4-cyanophenol to the anhydrous ethanol solution of hydrogen chloride is 1:1 to 1:10, preferably 1:5 to 1:10, further preferably 1:8 to 1:10, more preferably 1:10; for the alcoholysis reaction, the reaction temperature is -10 to 20 °C, preferably -10 to 10 °C, further preferably -5 to 5 °C, still further preferably 0 to 5 °C, most preferably 2 °C, and the reaction time is 24 to 72 h, preferably 24 to 48 h, further preferably 36 to 48 h, still further preferably 36 to 44 h, most preferably 40 h.

[0023] Among them, the solvent used in the alcoholysis reaction is selected from anhydrous ethanol, acetonitrile, acetone, tetrahydrofuran, dioxane, N,N-dimethylformamide or 1,2-dichloroethane, preferably 1,2-dichloroethane. The amount of the solvent used is such that the raw materials in the system can be dissolved and the viscosity is moderate.

[0024] In some embodiments, the base is any one or a combination of two of inorganic bases and organic bases, preferably an organic base; the inorganic base is sodium carbonate or potassium carbonate; the organic base is triethylamine, lithium diisopropylamide or N,N-diisopropylethylamine, preferably triethylamine; the molar ratio of the ethyl 4-hydroxybenzimidate hydrochloride intermediate to the base and ethyl cyanoacetate is 1:1.1:1 to 1:5:10, preferably 1:1.1:1 to 1:2:5, more preferably 1:1.1:1 to 1:1.5:2, still more preferably 1:1.1:1 to 1:1.3:1.5, and most preferably 1:1.2:1; for the condensation reaction, the reaction temperature is 75-150 °C, preferably 75-120 °C, more preferably 75-100 °C, still more preferably 75-85 °C, and most preferably 80 °C.

[0025] Among them, the solvent used in the condensation reaction is selected from toluene, tetrahydrofuran, dioxane, N,N-dimethylformamide, dimethyl sulfoxide or anhydrous ethanol, preferably anhydrous ethanol. The amount of the solvent used is such that the raw materials in the system can be dissolved and the viscosity is moderate.

[0026] Among them, TLC is used for monitoring during the process of the condensation reaction, and the reaction time is until the raw materials in the reaction system react completely.

[0027] The application of the above-mentioned 2-cyanoacrylate compounds in the preparation of derivatives of 2-cyanoacrylate compounds is also within the protection scope of the present invention;

[0028] The structural formula of the derivative of the 2-cyanoacrylate compound is shown in Formula II:

[0029]

[0030] Among them, R = Na or K.

[0031] The present invention also discloses a derivative of a 2-cyanoacrylate compound, and the structural formula of the derivative of the 2-cyanoacrylate compound is shown in Formula II:

[0032]

[0033] Among them, R = Na or K.

[0034] Furthermore, the present invention discloses a preparation method of a derivative of the above-mentioned 2-cyanoacrylate compound. The 2-cyanoacrylate compound I reacts to form a salt under the action of a base, thereby obtaining the derivative II of the 2-cyanoacrylate compound;

[0035]

[0036] wherein, R = Na or K.

[0037] In some embodiments, the base is sodium hydroxide or potassium hydroxide; the molar ratio of the 2-cyanoacrylate compound I to the base is 1:1 to 1:20, preferably 1:1 to 1:15, more preferably 1:1 to 1:10, still more preferably 1:2 to 1:8, and most preferably 1:5; for the reaction to form a salt, the reaction temperature is 40 to 80 °C, preferably 40 to 75 °C, more preferably 40 to 60 °C, still more preferably 45 to 55 °C, and most preferably 50 °C, and the reaction time is 24 to 72 h, preferably 24 to 60 h, more preferably 36 to 60 h, still more preferably 42 to 54 h, and most preferably 48 h.

[0038] Among them, the solvent used in the reaction to form a salt is selected from acetonitrile, acetone, toluene, tetrahydrofuran, dioxane, N,N-dimethylformamide, dimethyl sulfoxide or absolute ethanol, preferably absolute ethanol, and the usage amount of the solvent is such that the raw materials in the system can be dissolved and the viscosity is moderate.

[0039] Finally, the present invention discloses a pesticide formulation, which contains 0.1% - 99.99% by weight of the above-mentioned 2-cyanoacrylate compound or the derivative of the above-mentioned 2-cyanoacrylate compound.

[0040] Specifically, the dosage form of the pesticide formulation is a suspension concentrate, a dispersible oil suspension, a dispersible concentrate, a suspension seed coating agent, a tablet, a microemulsion, an aqueous solution, an emulsion, an emulsifiable concentrate, a water suspension, a powder, a wettable powder, a soluble powder, a soluble concentrate, a granule, a soluble granule, a water-dispersible granule, a capsule, a microcapsule, a microcapsule suspension or a nano-formulation.

[0041] Specifically, the pesticide formulation contains agriculturally acceptable carriers and / or excipients.

[0042] Among them, the carrier can be liquid or solid. The liquid carrier includes water and organic solvents. When water is used as a solvent or diluent, the organic solvent can also be used as an auxiliary agent or an antifreeze additive. Suitable organic solvents include any one or a combination of several of aromatic hydrocarbons (such as benzene, toluene, xylene, etc.), aliphatic hydrocarbons (such as cyclohexane, petroleum fractions, light mineral oil, etc.), chlorinated hydrocarbons (such as chlorobenzene, chloroform, vinyl chloride, dichloromethane, etc.), ketones (such as cyclohexanone, acetone, dimethylformamide, N-methyl-pyrrolidone, etc.), ethers, esters and alcohols (such as isopropanol, ethylene glycol, butanol, glycerol, cyclohexanol, etc.). The solid carrier includes any one or a combination of several of natural or synthetic clays and silicates (such as diatomaceous earth, natural silica, etc.), magnesium aluminum silicate (such as kaolin, kaolinite, montmorillonite, mica, etc.), magnesium silicate (such as talc), limestone, calcium carbonate, white carbon black, light calcium carbonate, sodium sulfate, calcium sulfate and amine salts (such as hexamethylenediamine or ammonium sulfate).

[0043] Among them, the carrier can also be a surfactant. Suitable surfactants can be emulsifiers (ionic or non-ionic), dispersants or wetting agents. Among them, non-ionic emulsifiers such as polyoxyethylene fatty alcohol ethers, polyoxyethylene fatty acid esters, polyoxyethylene fatty amines and commercially available emulsifiers (including agricultural emulsifier 100#, agricultural emulsifier 500#, agricultural emulsifier 600#, agricultural emulsifier 600-2#, agricultural emulsifier 2201B, agricultural emulsifier NP-10, agricultural emulsifier 36#, agricultural emulsifier 0203B, agricultural emulsifier 1601, agricultural emulsifier OX-667, agricultural emulsifier 2201, agricultural emulsifier NP-15, agricultural emulsifier 507#, agricultural emulsifier OX-635, agricultural emulsifier OX-622, agricultural emulsifier OX-653, etc.) in any one or a combination of several; dispersants include any one or a combination of several of methylnaphthalenesulfonic acid formaldehyde condensate, sodium lignosulfonate, calcium lignosulfonate and Nekal; wetting agents include any one or a combination of several of sodium alkylnaphthalenesulfonate, sodium lauryl sulfate and sodium dodecylbenzenesulfonate.

[0044] The application of the above-mentioned 2-cyanoacrylate compounds or derivatives of the above-mentioned 2-cyanoacrylate compounds or the above-mentioned pesticide formulations in controlling fungal diseases of crops is also within the protection scope of the present invention.

[0045] Specifically, in the above-mentioned application, the fungal diseases are diseases caused by any one or more of the genera Pyricularia, Ustilaginoidea, Fusarium, Gaeumannomyces, Colletotrichum, Cercospora, Verticillium, Valsa, Alternaria, Magnaporthe, Exserohilum, Rhizoctonia and Botrytis.

[0046] Specifically, in the application described above, the diseases are any one or more of rice blast, false smut of rice, Fusarium head blight of wheat, bakanae disease of rice, Fusarium oxysporum f. sp. cubense, basal stalk rot of wheat, Gaeumannomyces graminis var. tritici, anthracnose of strawberry, anthracnose of apple, anthracnose of grape, anthracnose of osmanthus, anthracnose of citrus, anthracnose of rubber tree, anthracnose of maize, anthracnose of soybean, anthracnose of scallion, anthracnose of pepper, anthracnose of wolfberry, anthracnose of mango, anthracnose of camellia, anthracnose of yam, brown spot of peanut, Verticillium wilt of cotton, apple tree canker, early blight of tomato, summer leaf spot, northern leaf blight of maize, sheath blight of wheat, and gray mold of fruits and vegetables.

[0047] Specifically, in the application described above, a 2-cyanoacrylate compound or its derivative or a pesticide formulation is applied to the pathogen to be controlled or the medium for its growth, and the dosage is 0.001 - 1000 g / ha, preferably 1 - 1000 g / ha, more preferably 20 - 500 g / ha, and even more preferably 20 - 320 g / ha; wherein, the dosage is calculated based on the mass of the 2-cyanoacrylate compound.

[0048] Among them, preferably, the dosage of the 2-cyanoacrylate compound or its derivative is 1 - 1000 g / ha, more preferably, the dosage of the 2-cyanoacrylate compound or its derivative is 20 - 150 g / ha, even more preferably, the dosage of the 2-cyanoacrylate compound or its derivative is 20 - 100 g / ha, and most preferably, the dosage of the 2-cyanoacrylate compound or its derivative is 20 - 80 g / ha; wherein, the dosage is calculated based on the mass of the 2-cyanoacrylate compound.

[0049] Among them, preferably, the dosage of the pesticide formulation is 1 - 1000 g / ha (the dosage is calculated based on the mass of the pesticide formulation), more preferably, the dosage of the pesticide formulation is 100 - 500 g / ha, even more preferably, the dosage of the pesticide formulation is 100 - 320 g / ha, and most preferably, the dosage of the pesticide formulation is 120 - 280 g / ha.

[0050] Inventive concept: Previous studies found that the target of cyazofamid is myosin-5. By comparing the amino acid sequences of the ligand-binding pockets of myosin-5 from plant pathogenic fungi sensitive to cyazofamid (such as Fusarium graminearum, Fusarium asiaticum, Fusarium oxysporum, etc.) with those of myosin-5 from plant pathogenic fungi insensitive to cyazofamid (such as Magnaporthe oryzae, Ustilaginoidea virens, Colletotrichum spp., etc.), it was found that there was only one amino acid site difference, namely site 375. In the myosin-5 of sensitive fungi, the amino acid at site 375 is methionine (M), while in the myosin-5 of strains insensitive to cyazofamid, the corresponding amino acid at site 375 is lysine (K). We further analyzed the three-dimensional structures of myosin-5 from Fusarium graminearum and Magnaporthe oryzae and found that there were only structural differences at site 375 in the ligand-binding pocket cavity. In the myosin-5 of Fusarium graminearum, the amino acid at site 375 is methionine (M), while in the myosin-5 of Magnaporthe oryzae, the corresponding amino acid at site 375 is the amino acid at site 378, lysine (K), which has a different structural type from methionine. By mutating the amino acid at site 378 of Magnaporthe oryzae myosin-5 (K to M, K378M), it was found that cyazofamid could inhibit the ATPase activity of the point mutant protein, while cyazofamid had no inhibitory activity on the ATPase of the wild-type Magnaporthe oryzae myosin-5. The 50% effective concentration (EC 50 ) of cyazofamid against the mycelial growth of Magnaporthe oryzae was 77.08 μg / mL, while the EC 50 against the K378M Magnaporthe oryzae mutant was 0.25 μg / mL. The above results indicate that the mutation of methionine to lysine is the key reason for the insensitivity of plant pathogenic fungi to cyazofamid. Therefore, molecular groups were designed to form strong interactions with lysine in order to expand the antibacterial spectrum of 2-cyanoacrylate compounds. At the same time, the physicochemical properties of the compounds were changed in order to reduce the toxicity of 2-cyanoacrylate compounds to non-target organisms.

[0051] Beneficial effects:

[0052] (1) The novel 2-cyanoacrylate compounds I and the derivatives II of 2-cyanoacrylate compounds provided by the present invention have good bactericidal activities and have excellent bactericidal activities against the pathogens of Pyricularia, Ustilaginoidea, Fusarium, Gaeumannomyces, Colletotrichum, Cercospora, Verticillium, Valsa, Alternaria, Magnaporthe, Exserohilum, Rhizoctonia and Botrytis. The compounds can be widely used for preparing fungicides in the fields of agriculture, horticulture and industry, and have the advantages of broad-spectrum bactericidal, high efficiency, low toxicity and environmental friendliness.

[0053] (2) It has low toxicity to non-target organisms, good environmental compatibility, can be naturally and rapidly degraded in the environment, will not cause residual harm to the environment, and has high safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] The present invention will be further specifically described below in conjunction with the drawings and specific embodiments, and the above and / or other advantages of the present invention will become clearer.

[0055] Figure 1 It is the preventive effect diagram of compound No. 1 suspending agent on mango anthracnose. SPECIFIC EMBODIMENTS

[0056] The present invention will be further described in detail below in conjunction with examples and data, and the present invention is not limited to these specific embodiments. Those skilled in the art should recognize that the present invention covers all alternative, improved and equivalent solutions that may be included within the scope of the claims. In the present invention, unless otherwise specified, all parts and percentages are in weight units, and the equipment and raw materials used can be purchased from the market or are commonly used in the art. The methods in the following examples are conventional methods in the art unless otherwise specified.

[0057] Example 1: Preparation of 2-cyanoacrylate compound I

[0058] Weigh p-hydroxybenzonitrile (5 mmol, 0.595 g) into a 50 mL eggplant-shaped flask, add 2 mL of 1,2-dichloroethane as a solvent, and place it in a low-temperature reaction bath at 2 °C. Add anhydrous ethanol solution of hydrogen chloride (50 mmol, 4.130 g), and carry out alcoholysis reaction with low-temperature stirring for 40 h; after the reaction is completed, concentrate by reduced pressure distillation, let it stand and crystallize at low temperature to obtain the intermediate 4-hydroxybenzimidic acid ethyl ester hydrochloride; directly use it for the next reaction without purification.

[0059] Weigh the intermediate 4-hydroxybenzimidic acid ethyl ester hydrochloride (3 mmol, 0.605 g) into an eggplant-shaped flask, add 5 mL of anhydrous ethanol for dissolution, then place it in a low-temperature reaction bath at 5-10 °C, use triethylamine as an acid-binding agent, slowly dropwise add (Et3N, 3.6 mmol, 0.367 g), and continue stirring for 30 min; then slowly heat up to reflux (80 °C), slowly dropwise add ethyl cyanoacetate (3 mmol, 0.339 g), and after the addition, carry out reflux reaction until the condensation reaction is complete. Monitor the reaction by TLC. After the reaction is completed, remove the solvent by reduced pressure distillation, and carry out column chromatography with dichloromethane and methanol as eluents (DCM:MeOH = 60:1) to obtain 2-cyanoacrylate compound I, a white solid, with a yield of 78%.

[0060] The 1H NMR data of 2-cyanoacrylate compound I are as follows:

[0061] 1 1H NMR (300 MHz, DMSO-d6) δ 10.10 (s, 1H), 9.18 (s, 1H), 8.73 (s, 1H), 7.48 - 7.39 (m, 2H), 6.92 - 6.82 (m, 2H), 4.17 (q, J = 7.1 Hz, 2H), 1.30 - 1.19 (m, 3H).

[0062] The prepared 2-cyanoacrylate compound I was designated as Compound No. 1.

[0063] Example 2: Preparation of Derivative II of 2-Cyanoacrylate Compound

[0064] Preparation of derivative II (R = Na) of 2-cyanoacrylate compound: Weigh 2-cyanoacrylate compound I (1 mmol, 232 mg, prepared in Example 1) into a round-bottom flask, and then add sodium hydroxide (5 mmol, 200 mg). Using 10 mL of absolute ethanol as the solvent, react at 50 °C for 48 h to form a salt; after the reaction is completed, evaporate the solvent under reduced pressure to obtain derivative II of 2-cyanoacrylate compound (designated as Compound No. 2, R = Na), a white solid, with a yield of 90%.

[0065] The 1H NMR data of Compound No. 2 (R = Na) are as follows:

[0066] 1 1H NMR (500 MHz, DMSO-d6) δ 9.20 (s, 1H), 8.75 (s, 1H), 7.49–7.42 (m, 2H), 6.94–6.84 (m, 2H), 4.19 (q, J = 7.1 Hz, 2H), 1.26 (t, J = 7.1 Hz, 3H).

[0067] Preparation of derivative II (R = K) of 2-cyanoacrylate compound: Weigh 2-cyanoacrylate compound I (1 mmol, 0.232 g, prepared in Example 1) into a round-bottom flask, and then add potassium hydroxide (5 mmol, 0.281 g). Using 10 mL of absolute ethanol as the solvent, react at 50 °C for 48 h to form a salt; after the reaction is completed, evaporate the solvent under reduced pressure to obtain derivative II of 2-cyanoacrylate compound (designated as Compound No. 3, R = K), a white solid, with a yield of 90%.

[0068] The 1H NMR data of Compound No. 3 (R = K) are as follows:

[0069] 11H NMR (500 MHz, DMSO-d6) δ 9.26 (s, 1H), 8.99 (s, 1H), 7.67 - 7.61 (m, 2H), 7.35 - 7.29 (m, 2H), 4.22 (q, J = 7.1 Hz, 2H), 1.27 (d, J = 3.6 Hz, 3H).

[0070] In Examples 3 to 8 below, actual examples of formulating several fungicide formulations with the 2-cyanoacrylate compound I of the present invention as the active substance component are given. It should be noted that the present invention is not limited solely to the scope of the following examples. In these formulation examples, all "%" refers to weight percentage.

[0071] Example 3: Suspension Concentrate Formulation

[0072] Weigh 25% of Compound No.1 (prepared in Example 1), 2% of TERSPERSE 2500, 3% of TERSPERSE 2425, 0.25% of xanthan gum, 2.95% of silica white, 4.5% of ethylene glycol, 0.3% of benzoic acid, 0.5% of silicone defoamer (trade name: s-29 produced by Nanjing Sixin Applied Chemicals Co., Ltd.), and add deionized water to 100% mass parts. According to the above formulation ratio, using water as the medium, add the technical material, dispersant, suspending agent, antifreeze agent, etc. into the batching kettle and mix evenly, then mill for 35 min by a ball mill or high-speed shear dispersion, and then mill with a sand mill to prepare.

[0073] Example 4: Wettable Powder Formulation

[0074] Mix 20% of Compound No.1 (prepared in Example 1), 5% of lignosulfonate, 1% of lauryl alcohol polyoxyethylene ether (JFC), and 38% of light calcium carbonate, and make up to 100% with diatomaceous earth, then mix evenly and pulverize to obtain the wettable powder.

[0075] Example 5: Emulsifiable Concentrate Formulation

[0076] Heat 15% of Compound No.1 (prepared in Example 1), 5% of agricultural emulsifier 500 (calcium salt), 5% of agricultural emulsifier 602, 5% of N-methyl-2-pyrrolidone, and 70% of xylene to 80 °C and stir evenly to obtain the emulsifiable concentrate.

[0077] Example 6: Water Dispersible Granule Formulation

[0078] Mix 30% of Compound No.1 (prepared in Example 1), 4% of naphthalene sulfonate formaldehyde condensate, 1% of naphthalene sulfonate, 2% of silica white, and 63% of kaolin, then pulverize, add water to knead, add to a granulator equipped with a certain specification sieve mesh for granulation, and then dry and screen (according to the sieve mesh range) to obtain the water dispersible granule product.

[0079] Example 7: Granule formulation

[0080] 10% of Compound No. 1 (prepared in Example 1), 1% of polyvinyl alcohol (PVA), 4% of naphthalene sulfonate formaldehyde condensate (NMO), and 85% of clay were uniformly mixed and pulverized. Then, 20 parts of water were added to 100 parts of this mixture, kneaded, and granulated into 14 - 32 mesh granules using an extrusion granulator, and then dried to obtain the granule formulation.

[0081] Example 8: Aqueous suspension formulation

[0082] 30% of Compound No. 1 (prepared in Example 1), 1% of fatty alcohol polyoxyethylene ether, 3% of rosin block polyoxyethylene ether polyoxypropylene ether sulfonate, 1% of magnesium aluminum silicate, 0.4% of silicone defoamer, 5% of propylene glycol, and deionized water (59.5%) were pre - mixed uniformly, then ground in a sand mill, and after filtration, a suspension mother liquor was obtained. An aqueous solution of xanthan gum (0.1%) prepared was added and shear - mixed uniformly to obtain the aqueous suspension formulation.

[0083] Examples of bioactivity assays using the compounds of the present invention are given below. It should be noted that the present invention is not limited solely to the scope of the following examples.

[0084] Example 9: Antibacterial activity assay

[0085] Compound No. 1 used in this example was prepared in Example 1, and Compound No. 2 (R = Na) and Compound No. 3 (R = K) were prepared in Example 2.

[0086] A PDA plate containing an equal volume of DMSO was used as a solvent control, and an equal - concentration of phenamacril was used as a control agent. The test pathogenic bacteria were inoculated on a PDA plate and cultured in a biochemical incubator at 25°C until the logarithmic phase. A puncher was used to punch out 5 - mm - diameter disks from the edge of the fresh colony and inoculated on a PDA plate containing a certain concentration of the drug to be tested. At the same time, the disks were inoculated onto PDA media without the drug to be tested and containing phenamacril as controls, and cultured in the dark in a biochemical incubator at 25°C until the control without the drug to be tested was nearly full - grown. The colony diameter was measured by the cross - method (subtracting the 5 - mm disk diameter). The mycelial growth inhibition rate (MGIR) was calculated using the following formula: MGIR% = [(C - N) / C] × 100%, where C is the colony diameter of the control group and N is the colony diameter of the treatment group. Each treatment in this experiment was set with three replicates, and the experiment was repeated twice.

[0087] The test pathogenic bacteria are Magnaporthe oryzae (Pyricularia), Ustilaginoidea virens (Ustilaginoidea), Fusarium graminearum Schwabe (Gibberella zeae, Fusarium), Fusarium moniliforme Sheld. (Fusarium), Fusarium oxysporum f. sp. cubense (Fusarium oxysporum, Fusarium), Pseudograminearum sp. (Pseudograminearum, Fusarium), Gaeumannomyces graminis var. tritici (Gaeumannomyces), Colletotrichum gloeosporioides Penz. (Colletotrichum), Colletotrichum gloeosporioides Penz. (Colletotrichum), Colletotrichum gloeosporioides Penz. (Colletotrichum), Colletotrichum gloeosporioides Penz. (Colletotrichum), Colletotrichum gloeosporioides Penz. (Colletotrichum), Colletotrichum gloeosporioides Penz. (Colletotrichum), Colletotrichum gloeosporioides Penz. (Colletotrichum), Colletotrichum gloeosporioides Penz. (Colletotrichum), Colletotrichum gloeosporioides Penz. (Colletotrichum), Colletotrichum gloeosporioides Penz. (Colletotrichum), Cercospora arachidicola Hori (Cercospora), Verticillium dahliae Kleb. (Verticillium), Valsa mali Miyabe et Yamada (Valsa), Alternaria solani Sorauer (Alternaria), Pyricularia oryzae Cav. (Pyricularia), Exserohilum turcicum (Pass.) Leonard & Suggs (Exserohilum), Rhizoctonia solani Kühn (Rhizoctonia) and Botrytis cinerea Pers. (Botrytis). Except that the test concentration of the agents used for Magnaporthe oryzae and Fusarium graminearum is 1 μg / mL, the test concentration of the agents used for the rest of the pathogenic bacteria is 10 μg / mL. "mg / L" refers to per milligram of active substance / liter, and fenoxanil is used as the control agent.

[0088] Table 1 Mycelial growth inhibition rate of 2-cyanoacrylate compounds against phytopathogenic fungi

[0089]

[0090]

[0091]

[0092] Table 2 Indoor toxicity determination of compound No.1 against some pathogenic bacteria

[0093]

[0094] The results in Table 1 and Table 2 show that: The compound prepared by the present invention has good bactericidal activity. It not only has good bactericidal activity against Fusarium fungi, but also has outstanding bactericidal activity against other phytopathogenic fungi, such as the pathogenic bacteria of rice blast, false smut of rice, anthracnose, etc. However, the 2-cyanoacrylate compound fenoxanil has no inhibitory activity or very poor inhibitory activity on the mycelial growth of the above-mentioned pathogenic bacteria. Therefore, the compound of the present invention significantly expands the control spectrum of the agent.

[0095] Example 10: Preventive and therapeutic effects of compound No.1 on rice blast

[0096] The compound used in this example was prepared in Example 1 of Compound No. 1.

[0097] Determination of protective effect: After puncturing holes at the bottom of disposable plastic cups (350 mL), add soil mixed by black soil and vermiculite at a ratio of 2:1. Put about 20 rice seeds in each cup, cover the seeds with a layer of soil, and place them in a greenhouse (22 - 25 °C) for 7 days (1-leaf stage) for use. Adding medicine: To ensure that the amount of medicine absorbed by the leaves is the same, in this experiment, the leaves were cut and placed in a sterile plastic dish, and a unified amount of medicine was added to the dish. The test agent was prepared into the required concentration, 2 mL of the liquid medicine was added to each dish, and after placing it in the greenhouse for 24 hours, inoculation was carried out. The control group was a water control. This experiment used the drop method for inoculation. After inoculating the spore solution on the leaves, first place the plastic dish containing the leaf segments at 28 °C, without light, and a relative humidity of 80% for 24 hours, then change the dish to an alternating light and dark (12 hours of light - 12 hours of darkness) condition at 28 °C, keep the leaf segments moist, and after five days, take out the dish and record the length of the disease spots. And calculate the disease index and relative control effect according to the following formula:

[0098] The classification of resistance and susceptibility follows the grading standard of 0 - 5 levels. Level 0: No disease spots at all; Level 1: Brown dot disease spots with a diameter not exceeding 0.5 mm; Level 2: Brown dot disease spots with a diameter of 0.5 - 1 mm; Level 3: Oval disease spots with a diameter of 1 - 3 mm, brown around and grayish-white in the center; Level 4: Typical spindle-shaped disease spots with a diameter of 3 mm or longer, with the disease spots slightly fused or not fused; Level 5: The same as Level 4, but due to the fusion of the disease spots, the upper half of the leaf withers.

[0099]

[0100]

[0101] Determination of therapeutic effect: After puncturing holes at the bottom of disposable plastic cups (350 mL), add soil mixed by black soil and vermiculite at a ratio of 2:1. Put about 20 rice seeds in each cup, cover the seeds with a layer of soil, and place them in a greenhouse (22 - 25 °C) for 7 days for use. This experiment used the drop method for inoculation. After inoculating spores on the leaves, first place the plastic dish containing the leaf segments at 28 °C, without light, and a relative humidity of 80% for 24 hours, then add medicine to the dish and change it to an alternating light and dark (12 hours of light - 12 hours of darkness), keep the leaf segments moist, and after five days, take out the dish and record the length of the disease spots. 24 hours after inoculating the leaf segments with the spore solution, the test agent was prepared into the required concentration, 2 mL of the liquid medicine was added to each dish (evenly added between the leaves), and after adding, put it back in the greenhouse to continue the disease development. The control group was a water control. And calculate the disease index and relative control effect according to the above formula.

[0102] The results in Table 3 and Table 4 show that the compound of the present invention has a significant preventive effect on rice blast, and the preventive effect is significantly higher than that of the control agents isoprothiolane and cyenopyrafen. At the same time, the compound of the present invention has a certain therapeutic effect on rice blast.

[0103] Table 3 Preventive effect of Compound No.1 against Magnaporthe oryzae

[0104]

[0105] Table 4 Therapeutic effect of Compound No.1 against Magnaporthe oryzae

[0106]

[0107] Example 11: Preventive effect of Compound No.1 against Fusarium head blight of wheat

[0108] Compound No.1 used in this example was prepared in Example 1.

[0109] Compound No.1 was diluted with sterile water containing 0.2% Tween 20 into medicament solutions of 5 μg / mL, 50 μg / mL, 250 μg / mL and 500 μg / mL. The fenaminstrobin was diluted to the same concentration as the positive control, and sterile water containing 0.2% Tween 20 was used as the negative control. The middle sections of wheat leaves with similar growth conditions were selected for marking and coated with the medicament solutions at different concentrations, and there were 6 replicates for each medicament concentration. 3 - 4 h after coating, the wild - type strain PH - 1 of Fusarium graminearum was inoculated at the coated sites, and the fungus was also inoculated at the uncoated sites on both sides of the marks to determine the conductivity of the medicament. The inoculated parts were wrapped with absorbent cotton to keep them moist. The wheat was cultured under the conditions of 21°C, a photoperiod of 12 h, and a relative humidity of 90%. After 3 d, the length of the lesion was measured. And the relative control efficacy was calculated according to the following formula.

[0110]

[0111] The results in Table 5 show that Compound No.1 of the present invention has a significant preventive effect against Fusarium head blight of wheat.

[0112] Table 5 Preventive effect of Compound No.1 against Fusarium head blight of wheat

[0113]

[0114]

[0115] Example 12: Preventive effect of the suspending agent of Compound No.1 against Colletotrichum gloeosporioides of mango

[0116] In the suspension agent ratio used in this example, the percentage contents are all mass percentages, and the preparation method is the same as that in Example 3: Weigh 25% of Compound No.1 (prepared in Example 1), 2% of TERSPERSE 2500, 3% of TERSPERSE 2425, 0.25% of xanthan gum, 2.95% of silica white, 4.5% of ethylene glycol, 0.3% of benzoic acid, 0.5% of silicone defoamer (trade name: s-29 produced by Nanjing Sixin Applied Chemicals Co., Ltd.), and add deionized water to 100% by mass. According to the above formula ratio, using water as the medium, add the technical material, dispersant, suspending agent, antifreeze agent, etc. into the batching kettle and mix evenly, grind for 35 min by a ball mill or high-speed shear dispersion, and then grind with a sand mill to prepare it.

[0117] After transferring and subculturing Colletotrichum gloeosporioides, it is reserved for use; Prepare the 25% suspension agent of Compound No.1 into aqueous solutions containing the drug with concentrations of 0 μg / mL, 1 μg / mL, 10 μg / mL, 100 μg / mL, and 200 μg / mL with 0.2% Tween water, 5 mL for each concentration. The control agent is the technical material of prochloraz, and the application concentration is 100 μg / mL; A total of 6 treatments are set in the experiment, and each treatment has 3 replicates, and the experiment is repeated three times. Select 6 mango fruits with similar growth conditions as a group, and divide them into 3 groups in total; Immerse the mangoes in 2% sodium hypochlorite solution for 1 minute, then rinse and dry them with clear water and ddH2O successively; Pierce 3 wounds (about 3 mm deep) on each mango fruit with a pipette tip at appropriate intervals, inject 10 μL of the liquid medicine at each wound, and wait for the liquid medicine to be completely absorbed, then apply the cultured Colletotrichum gloeosporioides cake on the wound; Take an appropriate amount of absorbent cotton, moisten it with water, cover it on the cake, and wrap the mango fruit with plastic wrap. Remove the absorbent cotton and plastic wrap after 2 d; Place each group of fruits in a paper box, seal it with a transparent plastic bag, add an appropriate amount of wet paper towels and spray water on time to keep the humidity at about 95%, place it in a 28 °C plant incubator, take pictures to record the lesion situation every 2 d and measure the lesion diameter (mm). The experiment is repeated twice.

[0118]

[0119] Figure 1 The results in Table 6 show that the compound preparation of the present invention has a significant preventive effect on anthracnose.

[0120] Table 6 Preventive effect of Compound No.1 on mango anthracnose

[0121]

[0122] Example 13: Field efficacy test of 25% suspension agent of Compound No.1 against rice blast

[0123] The percentages in the suspension agent ratio used in this example are all mass percentages, and the preparation method is the same as that in Example 3: 25% of compound No. 1 (prepared in Example 1), 2% of TERSPERSE 2500, 3% of TERSPERSE 2425, 0.25% of xanthan gum, 2.95% of white carbon black, 4.5% of ethylene glycol, 0.3% of benzoic acid, 0.5% of organosilicon defoamer (trade name: s-29 produced by Nanjing Sixin Applied Chemicals Co., Ltd.), and deionized water is added to 100% by mass. According to the above formula ratio, with water as the medium, the original drug, dispersant, suspending agent and antifreeze agent are added to the batching kettle and mixed evenly, dispersed by ball mill or high-speed shearing for 35 minutes, and sanded by sand mill to prepare.

[0124] The tested rice was the rice blast susceptible variety Taihu Nuo No. 2. Before application, the rice was growing well, and the rice blast in the field was mainly panicle blast grade 2. The field planting and management measures of each experimental plot were consistent. The second application was carried out 7 days after the first application, for a total of 2 applications. 25 days after the second application, the incidence of rice blast was investigated according to the "GB / T 15790-2009 Specification for Survey and Investigation of Rice Blast Disease", and the control effect was calculated according to the following formula.

[0125]

[0126]

[0127] The results in Table 7 show that the compounds of the present invention have excellent control effects on rice blast in the field.

[0128] Table 7 Field efficacy of 25% suspension concentrate of compound No.1 against rice blast

[0129]

[0130] Example 14: Acute toxicity of compound No. 1 to zebrafish

[0131] Compound No. 1 used in this example was prepared according to Example 1.

[0132] Weigh compound No.1, the control agent 2-cyano-3-amino-3-phenyl acrylate (dissolved in DMSO), and the experimental reference agent potassium dichromate (dissolved in water) separately to prepare a stock solution with a concentration of 10,000.0 mg / L. After ultrasonication for 30 s with an ultrasonic cleaner, perform gradient dilution of the test solution and transfer it to a round glass fish tank. Add 10 zebrafish to each test fish tank and culture them in a determination chamber (23 °C, light-dark ratio 16 h:8 h), and observe and record the number of dead zebrafish and poisoning symptoms 96 h after exposure to the toxin. Do not feed during the test. At the start of the test, measure the temperature, pH value, dissolved oxygen content, light intensity, and water quality hardness at each treatment concentration. During the test, measure the temperature, pH value, dissolved oxygen content, and water quality hardness of the solution every 24 h. During the test, measure the ratios of the temperature, pH value, and dissolved oxygen content of each concentration solution at 0 h, 24 h, 48 h, 72 h, and 96 h respectively. The criterion for judging the death of zebrafish: Gently touch the tail of the fish with a glass rod, and if there is no response from the fish body, it is considered dead.

[0133] Observe and record the number of dead zebrafish and poisoning symptoms, environmental temperature, and light intensity 96 h after exposure to the toxin, and record the pH, dissolved oxygen, and water quality hardness at the start and end of the experiment. For the semi-static method, measure the content before and after replacing the test substance solution, and the content needs to be detected for each concentration group. Sample from the middle of the water body, and after sampling, pass through a 0.45 μm filter membrane to remove insoluble substances. When the test data shows that the concentration change of the test substance during the test does not exceed 20% of the designed concentration or the initial concentration, the LC can be calculated based on the designed concentration or the initial concentration. 50 When the test data shows that the concentration change of the test substance during the test exceeds 20% of the designed concentration or the initial concentration, calculate the LC based on the measured concentration. 50 Use the statistical software DPS 7.05 for analysis and calculation.

[0134] Quality control: During the domestication period, the mortality rate of zebrafish does not exceed 5%; during the test, the mortality rate of the blank control does not exceed 10% and the laboratory conditions are normal; the acute toxicity result LC of potassium dichromate to zebrafish 50 (24 h) is in the range of 200 - 400 mg / L. According to the LC value (96 h) (mg / L) of fish semi-lethal concentration, in accordance with the national standard "Test Guidelines for Environmental Safety Assessment of Chemical Pesticides - Part 12 Acute Toxicity Test for Fish" (GB / T 31270.12 - 2014), the toxicity level of pesticides to fish is divided into four levels: highly toxic, LC50 is toxic to fish; highly toxic, 0.1 mg / L < LC 50 ≤1.0 mg / L; moderately toxic, 1.0 mg / L < LC 50 ≤10 mg / L; low toxic, LC 50 > 10 mg / L. 50

[0135] As shown in Table 8, the LC of potassium dichromate 50 is 318.6 mg / L, and the LC of fenaminstrobin 50 is 18.3 mg / L. The LC of Compound No.1 50 is 63.2 mg / L. The toxicity of Compound No.1 to zebrafish is low and significantly lower than that of the control agent fenaminstrobin.

[0136] Table 8 LC determination of different compounds on zebrafish 50

[0137] Compound <![CDATA[(LC 50 , mg / L)]]> Potassium dichromate (reference agent) 318.60 Jingangmycin (control agent) 18.30 Compound No.1 63.20

[0138] The present invention provides a 2-cyanoacrylate compound, a preparation method thereof, and ideas and methods for its application in pesticides. There are many specific methods and ways to implement this technical solution. The above description is only a preferred embodiment of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention. Each component not clearly defined in this embodiment can be realized by existing technologies.​

Claims

1. A 2-cyanoacrylate compound, characterized in that, The structural formula of the 2-cyanoacrylate compound is shown in Formula I:

2. The preparation method of the 2-cyanoacrylate compound according to claim 1, characterized in that, 4-Cyanophenol undergoes alcoholysis reaction with anhydrous ethanol solution of hydrogen chloride to obtain the intermediate ethyl 4-hydroxybenzimidate hydrochloride; subsequently, the intermediate ethyl 4-hydroxybenzimidate hydrochloride undergoes a condensation reaction with ethyl cyanoacetate under the action of a base to obtain the 2-cyanoacrylate compound I; 3. The preparation method according to claim 2, wherein The molar ratio of the 4-cyanophenol to the anhydrous ethanol solution of hydrogen chloride is 1:1 to 1:10; for the alcoholysis reaction, the reaction temperature is -10 to 20 °C, and the reaction time is 24 to 72 h.

4. The preparation method according to claim 2, characterized in that, The molar ratio of the 4-cyanophenol to the anhydrous ethanol solution of hydrogen chloride is 1:5 to 1:

10.

5. The preparation method according to claim 2, characterized in that, For the alcoholysis reaction, the reaction temperature is -10 to 10 °C, and the reaction time is 24 to 48 h.

6. The preparation method according to claim 2, characterized in that, The base is any one or a combination of two of inorganic bases and organic bases; the inorganic base is sodium carbonate or potassium carbonate; the organic base is triethylamine, lithium diisopropylamide or N,N-diisopropylethylamine; the molar ratio of the intermediate ethyl 4-hydroxybenzimidate hydrochloride to the base and ethyl cyanoacetate is 1:1.1:1 to 1:5:10; for the condensation reaction, the reaction temperature is 75 to 150 °C.

7. The preparation method according to claim 2, characterized in that, The base is an organic base; the organic base is triethylamine.

8. The preparation method according to claim 2, wherein, The molar ratio of the intermediate ethyl 4-hydroxybenzimidate hydrochloride to the base and ethyl cyanoacetate is 1:1.1:1 to 1:2:

5.

9. The preparation method according to claim 2, characterized in that, For the condensation reaction, the reaction temperature is 75 to 120 °C.

10. Use of the 2-cyanoacrylate compound according to claim 1 in the preparation of derivatives of 2-cyanoacrylate compounds; The structural formula of the derivative of the 2-cyanoacrylate compound is shown in Formula II: Among them, R = Na or K.

11. The derivative of the 2-cyanoacrylate compound according to claim 1, characterized in that, The structural formula of the derivative of the 2-cyanoacrylate compound is shown in Formula II: Wherein, R = Na or K.

12. A method for preparing the derivative according to claim 11, characterized in that, The 2-cyanoacrylate compound I reacts to form a salt under the action of a base to obtain the derivative II of the 2-cyanoacrylate compound; Wherein, R = Na or K.

13. The preparation method according to claim 12, characterized in that, The base is sodium hydroxide or potassium hydroxide; the molar ratio of the 2-cyanoacrylate compound I to the base is 1:1 to 1:20; for the reaction to form a salt, the reaction temperature is 40 to 80 °C, and the reaction time is 24 to 72 h.

14. The preparation method according to claim 12, wherein, The base is sodium hydroxide or potassium hydroxide; the molar ratio of the 2-cyanoacrylate compound I to the base is 1:1 to 1:

15.

15. The preparation method according to claim 12, wherein, For the reaction to form a salt, the reaction temperature is 40 to 75 °C, and the reaction time is 24 to 60 h.

16. A pesticide formulation, characterized in that, The pesticide formulation contains 0.1% - 99.99% by weight of the 2-cyanoacrylate compound according to claim 1 or the derivative of the 2-cyanoacrylate compound according to claim 11.

17. The pesticide formulation according to claim 16, characterized in that, The dosage form of the pesticide formulation is suspension concentrate, dispersible oil suspension, dispersible concentrate, suspension seed coating agent, tablet, microemulsion, aqueous solution, emulsion, emulsifiable concentrate, aqueous suspension, powder, wettable powder, soluble powder, soluble concentrate, granule, soluble granule, water dispersible granule, capsule, microcapsule, microcapsule suspension or nano formulation.

18. The pesticide preparation according to claim 16, characterized in that, The pesticide formulation contains agriculturally acceptable carriers and / or adjuvants.

19. Use of the 2-cyanoacrylate compound according to claim 1, or a derivative of the 2-cyanoacrylate compound according to claim 11, or the pesticide formulation according to claim 16, in controlling fungal diseases of crops.

20. The application according to claim 19, characterized in that The fungal diseases are diseases caused by any one or more of the genera Pyricularia, Ustilaginoidea, Fusarium, Gaeumannomyces, Colletotrichum, Cercospora, Verticillium, Valsa, Alternaria, Magnaporthe, Exserohilum, Rhizoctonia, and Botrytis.

21. The application according to claim 19, wherein The diseases are any one or more of the diseases such as rice blast, false smut of rice, scab of wheat, bakanae disease of rice, banana wilt, wheat basal rot, take-all of wheat, strawberry anthracnose, apple anthracnose, grape anthracnose, osmanthus anthracnose, citrus anthracnose, rubber tree anthracnose, corn anthracnose, soybean anthracnose, onion anthracnose, pepper anthracnose, wolfberry anthracnose, mango anthracnose, camellia anthracnose, yam anthracnose, peanut brown spot, cotton verticillium wilt, apple tree canker, tomato early blight, summer leaf spot, corn northern leaf blight, wheat sheath blight, and fruit and vegetable gray mold.

22. The application according to claim 19, wherein Applying the 2-cyanoacrylate compound or its derivative or the pesticide formulation to the pathogen to be controlled or the medium for its growth, with the dosage being 0.001 - 1000 g / ha.

23. The application according to claim 19, characterized in that Applying the 2-cyanoacrylate compound or its derivative or the pesticide formulation to the pathogen to be controlled or the medium for its growth, with the dosage being 1 - 1000 g / ha.

24. The application according to claim 19, characterized in that, Applying the 2-cyanoacrylate compound or its derivative or the pesticide formulation to the pathogen to be controlled or the medium for its growth, with the dosage being 20 - 500 g / ha.

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