A class of compounds containing trifluoromethyl oxadiazole and spiro structure, preparation method and application

Abstract

This invention relates to a compound containing trifluoromethyloxadiazole and a spirocyclic structure, or a pesticide-acceptable salt, hydrate, or solvate thereof, characterized in that the structure of the compound containing trifluoromethyloxadiazole and a spirocyclic structure is shown in general formula I. Compared with the prior art, the compound provided by this invention can be used to prepare histone deacetylase inhibitors, effectively inhibiting the growth and reproduction of various rust fungal spores, with effects comparable to fluopyram.

Description

Technical Field

[0001] This invention relates to the field of agricultural fungicides, specifically to a class of compounds containing trifluoromethyloxadiazole and spirocyclic structures, their preparation methods, and applications. Background Technology

[0002] Rust fungi are a common group of plant pathogenic fungi, with over 8,000 species discovered worldwide. Rust fungi are diverse, spread rapidly, and have a wide range of applications, causing diseases in multiple parts of over a thousand crops, including soybeans, wheat, and corn. In the mid-20th century, wheat stripe rust broke out in my country; in the 1950s, this disease caused a reduction of approximately 40% in wheat yields in my country. Rust fungi are caused by *Russula beanica* (also known as *Russula spp.*). Phakopsora pachyrhizi Soybean rust, caused by rust, is widely distributed in major soybean-producing areas such as southern my country and the Huang-Huai-Hai Plain. In severe cases, it can cause large-scale death of soybean leaves, resulting in yield reductions of up to 30% and causing huge economic losses to agriculture. Rust seriously threatens the agricultural ecological balance and food security in my country and even the world.

[0003] Currently, the long-term and excessive application of traditional chemical pesticides has led to significant drug resistance in pathogenic fungi, making the development of novel drugs an urgent issue in this field. Histone deacetylase (HDAC) inhibitors can block gene expression and cell signaling in pathogens such as fungi and pests, interfering with fungal spore germination, mycelial growth, and differentiation of infection structures. To address the problem of rust fungi developing resistance to existing pesticides, it is essential to further synthesize novel HDAC inhibitors with high fungicidal activity and new targets. Agricultural HDAC inhibitors, represented by TFMO, have become an important research direction in the creation of green pesticides due to their novel mechanism of action, high-efficiency fungicidal activity, and lack of cross-resistance with existing pesticides. Through continuous structural optimization, in-depth analysis of the mechanism of action, and development of field application technologies, these compounds are expected to provide a new generation of solutions for the control of major crop diseases such as rust, contributing to the sustainable development of agricultural production and the achievement of the goal of reducing pesticide use and increasing efficiency. Summary of the Invention

[0004] The purpose of this invention is to provide a class of compounds containing trifluoromethyloxadiazole and a spirocyclic structure, a preparation method, and applications. The compounds can be used to prepare histone deacetylase inhibitors, effectively inhibiting the growth and reproduction of various rust fungal spores, with effects comparable to fluopyram.

[0005] The objective of this invention can be achieved through the following technical solutions: One objective of this invention is to provide a compound containing trifluoromethyloxadiazole and a spirocyclic structure, or a pesticide-acceptable salt, hydrate, or solvate thereof, wherein the structure of the compound containing trifluoromethyloxadiazole and a spirocyclic structure is shown in general formula I: General Formula I Wherein, X is any one of ester group, carbonyl group, amide group or oxime ester group; M can be either a nitrogen atom or a carbon atom; Ring A is an unsubstituted saturated 4- to 6-membered ring; Ring B is a saturated 3- to 6-membered ring containing 0 to 2 heteroatoms, or a benzo[a]-fused ring containing 0 to 1 oxygen atom; R1 is located at The hydrogen, C1-C4 alkyl, C1-C4 alkoxy, cyano or halogen at any substitution position on the ring; R2 and R3 are each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, nitro, cyano, carbonyl, amino, methoxy, ethoxy, propylamino, isooxypropyl, n-butoxy, tert-butoxy, cyclohexyloxy, N-(N,N-dimethyl) substituent, N-(N,N-diethyl) substituent, N-(N,N-dipropyl) substituent, N-(N,N-dipropyl) substituent, N-(N, N-Dibutyl) substituent, N-(N-methyl-N-ethyl) substituent, N-(N-methyl-N-propyl) substituent, N-(N-methyl-N-butyl) substituent, N-(N-ethyl-N-propyl) substituent, N-(N-ethyl-N-butyl) substituent, N-(N-propyl-N-butyl) substituent, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 2 or 3 or 4-methylcyclohexyl, 2 or 3 or 4-ethylcyclohexyl, 2 or 3 or 4-hydroxycyclohexyl, 2 or 3 or 4-carboxycyclohexyl, 2 or 3 or 4-cyanocyclohexyl, 2 or 3 or 4-aminocyclohexyl, 2 or 3-tetrahydrofuranyl, 2 or 3-tetrahydrothiophenyl, N or 2 Or any one of 3-tetrahydropyrrole, 2, 3, or 4-tetrahydropyranyl, N, 2, 3, or 4-piperidinyl, N, 2, or 3-morpholinyl, N, 2-piperazinyl, N, 2, 3, or 3-(N-methylpiperazinyl), N, 2, 3, or 3-(N-benzylpiperazinyl)phenyl, 2, 3, or 4-methylphenyl, 2, 3, or 4-methoxyphenyl, 2, 3, or 4-aminophenyl, 2, 3, or 4-cyanophenyl, 2, 3, or 4-carboxyphenyl, 2, 3, or 4-nitrophenyl, 2, 3, or 4-hydroxyphenyl, naphthyl, 2, 3-furanyl, 2, 3-thienyl, N, 2, 3, or 3-pyrrole, 2, 3, or 4-pyridinyl, N, 2, or 3-(N-morpholinyl)phenyl, and indoleyl.

[0006] In this invention, ring A is an unsubstituted saturated 4- to 6-membered ring, meaning that ring A has no substituent groups at any position except for the R2 group.

[0007] In this invention, ring B is a substituted or unsubstituted saturated 3-6 membered ring containing 0-2 heteroatoms, or a benzo[a]-fused ring containing 0-1 oxygen atom. Ring B can be a saturated 3-6 membered ring containing 0-2 heteroatoms or a benzo[a]-fused ring containing 0-1 oxygen atom. When ring B is a saturated 3-6 membered ring containing 0-2 heteroatoms, the ring may optionally have other substituents besides the R3 group, or it may not have other substituents.

[0008] In this invention, multiple position numbers connected by "or" (such as "2 or 3 or 4-methylcyclohexyl") indicate that the substituent can be independently connected to any one of the listed atoms, which is a monosubstitution; for nitrogen-containing heterocycles, "N or X or Y-" indicates that the heterocyclic group can be connected by a nitrogen atom (N position) or by a carbon atom (X position, Y position) of the listed number.

[0009] Preferably, the structure of the compound containing trifluoromethyloxadiazole and a spirocyclic structure is shown in general formula II: General Formula II Where M is a nitrogen atom; X can be either a carbonyl group or an amide group; Ring A is an unsubstituted saturated 4- to 6-membered ring; Ring B is a substituted or unsubstituted saturated 3- to 6-membered ring containing 0 to 2 heteroatoms, or a benzo[a] heterocycle containing 0 to 1 oxygen atom; R1 is hydrogen; R2 and R3 are each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, nitro, cyano, carbonyl, amino, methoxy, ethoxy, propylamino, isooxypropyl, n-butoxy, tert-butoxy, cyclohexyloxy, N-(N,N-dimethyl) substituent, N-(N,N-diethyl) substituent, N-(N,N-dipropyl) substituent, N-(N,N-dipropyl) substituent, N-(N, N-Dibutyl) substituent, N-(N-methyl-N-ethyl) substituent, N-(N-methyl-N-propyl) substituent, N-(N-methyl-N-butyl) substituent, N-(N-ethyl-N-propyl) substituent, N-(N-ethyl-N-butyl) substituent, N-(N-propyl-N-butyl) substituent, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 2 or 3 or 4-methylcyclohexyl, 2 or 3 or 4-ethylcyclohexyl, 2 or 3 or 4-hydroxycyclohexyl, 2 or 3 or 4-carboxycyclohexyl, 2 or 3 or 4-cyanocyclohexyl, 2 or 3 or 4-aminocyclohexyl, 2 or 3-tetrahydrofuranyl, 2 or 3-tetrahydrothiophenyl, N or 2 Or any one of 3-tetrahydropyrrole, 2, 3, or 4-tetrahydropyranyl, N, 2, 3, or 4-piperidinyl, N, 2, or 3-morpholinyl, N, 2-piperazinyl, N, 2, 3, or 3-(N-methylpiperazinyl), N, 2, 3, or 3-(N-benzylpiperazinyl)phenyl, 2, 3, or 4-methylphenyl, 2, 3, or 4-methoxyphenyl, 2, 3, or 4-aminophenyl, 2, 3, or 4-cyanophenyl, 2, 3, or 4-carboxyphenyl, 2, 3, or 4-nitrophenyl, 2, 3, or 4-hydroxyphenyl, naphthyl, 2, 3-furanyl, 2, 3-thienyl, N, 2, 3, or 3-pyrrole, 2, 3, or 4-pyridinyl, N, 2, or 3-(N-morpholinyl)phenyl, and indoleyl.

[0010] More preferably, in general formula II, X is a carbonyl group; ring A is an unsubstituted saturated 4- to 5-membered ring; and ring B is an unsubstituted saturated 3- to 5-membered ring or a benzo[a]heterocyclic ring containing 0 to 1 oxygen atom.

[0011] Preferably, the structure of the compound containing trifluoromethyloxadiazole and a spirocyclic structure is shown in general formula III: Formula III Where M is a carbon atom; X is any one of ester, carbonyl, amide, or oxime ester; Ring A is an unsubstituted saturated 4- to 6-membered ring; Ring B is a substituted or unsubstituted saturated 3- to 6-membered ring containing 0 to 2 heteroatoms, or a benzo[a] heterocycle containing 0 to 1 oxygen atom; R1 is located at The hydrogen, C1-C4 alkyl, C1-C4 alkoxy, cyano or halogen at any substitution position on the ring; R2 and R3 are each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, nitro, cyano, carbonyl, amino, methoxy, ethoxy, propylamino, isooxypropyl, n-butoxy, tert-butoxy, cyclohexyloxy, N-(N,N-dimethyl) substituent, N-(N,N-diethyl) substituent, N-(N,N-dipropyl) substituent, N-(N,N-dipropyl) substituent, N-(N, N-Dibutyl) substituent, N-(N-methyl-N-ethyl) substituent, N-(N-methyl-N-propyl) substituent, N-(N-methyl-N-butyl) substituent, N-(N-ethyl-N-propyl) substituent, N-(N-ethyl-N-butyl) substituent, N-(N-propyl-N-butyl) substituent, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 2 or 3 or 4-methylcyclohexyl, 2 or 3 or 4-ethylcyclohexyl, 2 or 3 or 4-hydroxycyclohexyl, 2 or 3 or 4-carboxycyclohexyl, 2 or 3 or 4-cyanocyclohexyl, 2 or 3 or 4-aminocyclohexyl, 2 or 3-tetrahydrofuranyl, 2 or 3-tetrahydrothiophenyl, N or 2 Or any one of 3-tetrahydropyrrole, 2, 3, or 4-tetrahydropyranyl, N, 2, 3, or 4-piperidinyl, N, 2, or 3-morpholinyl, N, 2-piperazinyl, N, 2, 3, or 3-(N-methylpiperazinyl), N, 2, 3, or 3-(N-benzylpiperazinyl)phenyl, 2, 3, or 4-methylphenyl, 2, 3, or 4-methoxyphenyl, 2, 3, or 4-aminophenyl, 2, 3, or 4-cyanophenyl, 2, 3, or 4-carboxyphenyl, 2, 3, or 4-nitrophenyl, 2, 3, or 4-hydroxyphenyl, naphthyl, 2, 3-furanyl, 2, 3-thienyl, N, 2, 3, or 3-pyrrole, 2, 3, or 4-pyridinyl, N, 2, or 3-(N-morpholinyl)phenyl, and indoleyl.

[0012] More preferably, in general formula III, X is a carbonyl group; ring A is an unsubstituted saturated 4- to 5-membered ring; ring B is an unsubstituted saturated 3- to 5-membered ring or a benzo[a]heterocyclic ring containing 0 to 1 oxygen atom; and R1 is hydrogen.

[0013] Preferably, the structural formula of the compound containing trifluoromethyloxadiazole and a spirocyclic structure is selected from any one of the following tables: The second objective of this invention is to provide a method for preparing the compound containing trifluoromethyloxadiazole and a spirocyclic structure, the synthetic route of which is as follows: S1: In step S1, equimolar amounts of compound A and compound B are dissolved in an inert solvent and reacted at room temperature for 1 hour. The reaction process is monitored by TLC. After the reaction is completed, the inert solvent is removed by rotary evaporator. The obtained product does not need to be purified and can be used directly in the next step. S2: In step S2, equimolar amounts of compound C and compound D (TFAA trifluoroacetic anhydride) were dissolved in a solvent and reacted at room temperature for 3 hours. The reaction progress was monitored by TLC. After the reaction was completed, the solvent was removed by rotary evaporation and then purified by column chromatography to obtain intermediate E. The eluent used in the column chromatography was a mixed solution of petroleum ether (PE) and ethyl acetate (EA) with a volume ratio of 5:1. S3: In step S3, equimolar amounts of compound E and compound F were dissolved in a solvent, and N,N,N′,N′-tetramethylchloromethamidine hexafluorophosphate and N-methylimidazole were added. The reaction was carried out at room temperature for 2.5 h, and the reaction progress was monitored by TLC. After the reaction was completed, the solvent was removed by rotary evaporation, and then purified by column chromatography to obtain the target compound of general formula I. The eluent used in the column chromatography was a 1:1 volume ratio of petroleum ether (PE) / ethyl acetate (EA) mixed solution.

[0014] Preferably, in step S1, the inert solvent includes any one of tetrahydrofuran, diethyl ether, and toluene.

[0015] Preferably, in step S2, the solvent includes any one of tetrahydrofuran, diethyl ether, toluene, and N,N-dimethylformamide.

[0016] Preferably, in step S3, the solvent includes any one of tetrahydrofuran, toluene, dichloromethane, and N,N-dimethylformamide.

[0017] Preferably, in step S3, the molar ratio of N,N,N′,N′-tetramethylchloromethanemidazone hexafluorophosphate, N-methylimidazole, and compound E is 1:1.2, and the molar ratio of N-methylimidazole to compound E is 1:3.5.

[0018] A third objective of this invention is to provide the application of the compound containing trifluoromethyloxadiazole and a spirocyclic structure, or its pesticide-acceptable salt, hydrate, or solvate, in the preparation of histone deacetylase inhibitors.

[0019] Preferably, the compound containing trifluoromethyloxadiazole and a spirocyclic structure, or its phytochemically or pharmaceutically acceptable salts, hydrates, or solvates, can be used to prepare fungal or human histone deacetylase inhibitors.

[0020] More preferably, the compound containing trifluoromethyloxadiazole and a spirocyclic structure, or its pesticide-acceptable salt, hydrate, or solvate, can be used to prepare histone deacetylase inhibitors of plant pathogenic fungi.

[0021] The fourth objective of this invention is to provide the application of the compound containing trifluoromethyloxadiazole and a spirocyclic structure, or its agrochemically acceptable salt, hydrate, or solvate, in the control of plant fungal diseases.

[0022] Preferably, the plant fungal disease is a disease caused by any one or more of the following pathogens: soybean rust, corn rust, wheat leaf rust, wheat stripe rust, tomato gray mold, rapeseed sclerotinia, tomato early blight, and wheat stem rust.

[0023] The compounds containing trifluoromethyloxadiazole and spirocyclic structures, or their pesticide-acceptable salts, hydrates, or solvates provided by this invention, can effectively prevent and control crop rust diseases.

[0024] The fifth objective of this invention is to provide a pharmaceutical composition comprising a therapeutically effective amount of an active ingredient and a pharmaceutically and pesticide-acceptable carrier, wherein the active ingredient is at least one of the following: a compound containing a trifluoromethyloxadiazole and a spirocyclic structure as described above, a pharmaceutically and pesticide-acceptable salt thereof, or a pharmaceutically and pesticide-acceptable solvate thereof.

[0025] Compared with the prior art, the present invention has the following beneficial effects: (1) This invention provides a class of compounds containing trifluoromethyloxadiazole and spirocyclic structures, preparation methods and applications. The compounds can be used to prepare histone deacetylase inhibitors, which can effectively inhibit the growth and reproduction of various rust fungal spores, with effects comparable to fluoxetine.

[0026] (2) In this invention, the compound containing trifluoromethyloxadiazole and spirocyclic structure can be used to prevent and control plant fungal diseases caused by one or more of the following pathogens: soybean rust, corn rust, wheat leaf rust, wheat stripe rust, tomato gray mold, rapeseed sclerotinia, tomato early blight, and wheat stem rust.

[0027] (3) Experimental results show that the compound containing trifluoromethyloxadiazole and spirocyclic structure provided by the present invention has significant control effect on plant diseases caused by various rust pathogens, including soybean rust and corn rust, and its effect is better than that of conventional drug 30wt% flufenoxadiazam, and has the potential for further research and development. Attached Figure Description

[0028] Figure 1 The 1H NMR spectrum of compound 24; Figure 2 The carbon NMR spectrum of compound 24; Figure 3 The diagram shows the molecular docking results of compounds 24 and 57 of this invention. Detailed Implementation

[0029] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. These embodiments are based on the technical solution of the present invention and provide detailed implementation methods and specific operating procedures. However, the scope of protection of the present invention is not limited to the following embodiments.

[0030] Unless otherwise specified, the reagents, methods, instruments, and equipment used in this invention are conventional in the art. Unless otherwise specified, the reagents and materials used in the following examples are all commercially available.

[0031] Example 1: Preparation of (2-oxo-7-azaspiro[3.5]nonane-7-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone. Step 1: Preparation of (Z)-4-(N'-hydroxyaminocarbamoyl)benzoic acid Compound A (147.13 mg, 1 mmol) and compound B (69.49 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile. The mixture was reacted at room temperature for 1 h. After the reaction was completed by TLC monitoring, the solvent in the reaction solution was removed by rotary evaporation and dried to obtain 124 mg of white solid, namely compound C, with a yield of 65.1%.

[0032] Step 2: Preparation of 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid Compound C (180 mg, 1 mmol) and compound D (210 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile. The mixture was reacted at room temperature for 3 h. After the reaction was completed by TLC monitoring, the inert solvent in the reaction solution was removed by rotary evaporation. The mixture was then purified by column chromatography (eluent: PE: EA = 5:1) to obtain 180 mg of white solid, which is compound E, with a yield of 70.4%.

[0033] Step 3: Preparation of (2-oxo-7-azaspiro[3.5]nonane-7-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone Compound E (258 mg, 1 mmol) and compound F (127 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound 01 was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 68.2%.

[0034] Example 02: Preparation of (7-oxo-2-azaspiro[3.5]nonane-2-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 7-oxo-2-azaspiro[3.5]nonane (113 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidin hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 55.4%.

[0035] Example 03: Preparation of (2-oxa-6-azaspiro[3.3]hept-6-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl) methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 2-oxo-6-azaspiro[3.3]heptane (99.3 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 44.5%.

[0036] Example 04: Preparation of (3-oxo-9-azaspiro[5.5]undecane-9-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 3-oxo-9-azaspiro[5.5]undecane (155.4 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidin hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 56.4%.

[0037] Example 05: Preparation of (1,4-dioxo-8-azaspiro[4.5]decane-8-yl)(4-(5-(trifluoromethyl)-1,2,4-oxazoline-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 1,4-dioxa-8-azaspiro[4.5]decane (143.2 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol) and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 44.3%.

[0038] Example 06: Preparation of (5,8-dioxa-2-azaspiro[3,4]octane-2-yl)(4-(5-(trifluoromethyl)-1,2,4-oxazoline-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 5,8-dioxo-2-azaspiro[3,4]octane (115.3 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 66.3%.

[0039] Example 07: Preparation of (1,5-dioxa-9-azaspiro[5.5]undecane-9-yl)(4-(5-(trifluoromethyl)-1,2,4-oxazoline-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 1,5-dioxa-9-azaspiro[5.5]undecane (157.3 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 55.6%.

[0040] Example 08: Preparation of (5,9-dioxo-2-azaspiro[3.5]nonane-2-yl)(4-(5-(trifluoromethyl)-1,2,4-oxazol-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 5,9-dioxo-2-azaspiro[3.5]nonane (129.4 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 54.5%.

[0041] Example 09: Preparation of (1-oxo-5-azaspiro[2.3]hexane-5-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 1-oxo-5-azaspiro[2,3]hexane (85.23 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 43.4%.

[0042] Example 10: Preparation of (1,6-dioxaspiro[3.3]heptane-6-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 1-oxo-6-azaspiro[3.3]heptane (85.23 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 45.3%.

[0043] Example 11: Preparation of (6-oxo-2-azaspiro[3.4]octane-2-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 6-oxo-2-azaspiro[3,4]octane (114.3 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 65.3%.

[0044] Example 12: Preparation of (5-oxo-2-azaspiro[3.5]nonane-2-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 5-oxo-2-azaspiro[3.5]nonane (127.3 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidin hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 53.9%.

[0045] Example 13: Preparation of (2-oxo-6-azaspiro[3.4]octane-6-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 2-oxo-6-azaspiro[3,4]octane (113.5 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 54.2%.

[0046] Example 14: Preparation of (6-oxo-2-azaspiro[3.5]nonane-2-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 6-oxo-2-azaspiro[3.5]nonane (127.8 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidin hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed using a rotary evaporator, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 65.9%. Example 15: Preparation of (7-oxo-2-azaspiro[4.5]decane-2-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 7-oxo-2-azaspiro[4.5]decane (141.21 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 65.9%.

[0047] Example 16: Preparation of (1-oxo-7-azaspiro[3.5]non-7-yl)(4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)methyl ketone Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 1-oxo-7-azaspiro[3.5]nonane (127.21 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidin hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 53.4%.

[0048] Example 17: Preparation of 2-azaspiro[3.3]heptane-6-yl 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoate Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound spiro2-azaspiro[3.3]hepta-6-ol (113.3 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidin hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 45.6%.

[0049] Example 18: Preparation of (2-fluoro-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)(5-azaspiro[2.3]hexane-5-yl) methyl ketone Compound 2-fluoro-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (synthetic method referred to patent WO2021 / 175669, 2021) (276 mg, 1 mmol) and compound spiro5-azaspirane[2,3]hexane (83 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was obtained by column chromatography (eluent: PE: EA = 1:1). The target compound was a white solid with a yield of 54.2%.

[0050] Example 19: Preparation of (3-fluoro-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)(2-azaspiro[3.3]hept-2-yl)methyl ketone Compound 3-fluoro-4-(5-(trifluoromethyl)-1,2,4-oxazol-3-yl)benzoic acid (synthetic method according to patent WO2021 / 175669, 2021) (276 mg, 1 mmol) and compound 2-azaspiro[3.3]heptane (97 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was obtained by column chromatography (eluent: PE: EA = 1:1). The target compound was a white solid with a yield of 45.2%.

[0051] Example 20: Preparation of (3-chloro-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)(5-azaspiro[2.3]hexane-5-yl) methyl ketone Compound 3-chloro-4-(5-(trifluoromethyl)-1,2,4-oxazol-3-yl)benzoic acid (291 mg, 1 mmol) and compound 5-azaspiro[2,3]hexane (83 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was obtained by column chromatography (eluent: PE: EA = 1:1). The target compound was a white solid with a yield of 48.2%.

[0052] Example 21: Preparation of (3-bromo-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)(5-azaspiro[2.3]hexane-5-yl)methyl ketone Compound 3-bromo-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (synthetic method according to patent WO2021 / 175669, 2021) (335 mg, 1 mmol) and compound 5-azaspiro[2,3]hexane (83 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was obtained by column chromatography (eluent: PE: EA = 1:1). The target compound was a white solid with a yield of 58.2%.

[0053] Example 22: Preparation of (2-iodo-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)(5-azaspiro[2.3]hexane-5-yl)methyl Compound 2-iodo-4-(5-(trifluoromethyl)-1,2,4-oxazol-3-yl)benzoic acid (synthetic method according to patent WO2021 / 175669, 2021) (384 mg, 1 mmol) and compound 5-azaspiro[2,3]hexane (83 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was obtained by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 62.2%.

[0054] Example 23: Preparation of (2-methyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)(5-azaspiro[2.3]hexane-5-yl) methyl ketone Compound 2-methyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (synthetic method according to patent WO2021 / 175669, 2021) (272 mg, 1 mmol) and compound 5-azaspiro[2,3]hexane (83 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidin hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was obtained by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 62.2%.

[0055] Example 24: Preparation of (3-methoxy-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)(5-azaspiro[2.3]hexane-5-yl) methyl ketone Compound 3-methoxy-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (synthetic method according to patent WO2021 / 175669, 2021) (288 mg, 1 mmol) and compound 5-azaspiro[2,3]hexane (83 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol) and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was obtained by column chromatography (eluent: PE: EA = 1:1). The target compound was a white solid with a yield of 60.2%.

[0056] Example 25: Preparation of (2-ethyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)(5-azaspiro[2.3]hexane-5-yl) methyl ketone Compound 2-ethyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (synthetic method according to patent WO2021 / 175669, 2021) (286 mg, 1 mmol) and compound 5-azaspiro[2,3]hexane (83 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol) and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was obtained by column chromatography (eluent: PE: EA = 1:1). The target compound was a white solid with a yield of 67.2%.

[0057] Example 26: Preparation of (3-cyclopropyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)phenyl)(5-azaspiro[2.3]hex-5-yl) methyl ketone Compound 3-cyclopropyl-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (synthetic method according to patent WO2021 / 175669, 2021) (298 mg, 1 mmol) and compound 5-azaspiro[2,3]hexane (83 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol) and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was obtained by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 60.2%.

[0058] Example 27: Preparation of 2-(2-azaspiro[3.3]heptane-2-carboxyl)-5-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzonitrile Compound 2-cyano-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (synthetic method according to patent WO2021 / 175669, 2021) (283 mg, 1 mmol) and compound 2-azaspiroane[3.3]heptane (83 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was obtained by column chromatography (eluent: PE: EA = 1:1). The target compound was a white solid with a yield of 58.2%.

[0059] Example 28: Preparation of spiro[3.3]heptane-2-yl 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoate Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound spiro[3.3]hepta-2-ol (112.3 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was obtained by column chromatography with a PE:EA ratio of 1:1. The target compound was a white solid with a yield of 35.6%.

[0060] Example 29: Preparation of tert-butyl 2-((4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoyl)oxy)-7-azaspiro[3.5]nonane-7-carboxylic acid ester Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound tert-butyl-2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylic acid ester (241.5 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidin hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by column chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 35.6%.

[0061] Example 30: Preparation of tert-butyl 7-((4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoyl)oxy)-2-azaspiro[3.5]nonane-2-carboxylic acid ester Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound tert-butyl-7-hydroxy-2-azaspiro[3.5]nonane-2-carboxylic acid ester (241.5 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidin hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 35.6%.

[0062] Example 31: Preparation of N-(7-azaspiro[3.5]non-2-yl)-4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzamide Compound 4-(5-(trifluoromethyl)-1,2,4-oxadiazol-3-yl)benzoic acid (180 mg, 1 mmol) and compound 7-azaspiro[3.5]nonane-2-amine (140.3 mg, 1 mmol) were added to a 100 mL three-necked flask, followed by 20 mL of acetonitrile, N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol), and N-methylimidazole (3.5 mmol). The reaction was carried out at room temperature for 2.5 h. After the reaction was completed by TLC, the solvent in the reaction solution was removed by rotary evaporation, and the target compound was purified by chromatography (eluent: PE:EA = 1:1). The target compound was a white solid with a yield of 35.6%.

[0063] Other compounds can be synthesized using this method, with appropriate starting materials, and according to the general formula for synthetic reactions.

[0064] Table 1 shows the NMR data of compounds 01 to 107.

[0065] Table 1. Structures and NMR data of compounds containing trifluoromethyloxadiazole and spirocyclic structures. The compounds containing trifluoromethyloxadiazole and spirocyclic structures prepared in the embodiments of this invention all exhibit significant control effects against rust diseases caused by fungi. The inhibitory effects of the above-mentioned compound on plant pathogenic fungi are studied below.

[0066] Test Example: Experiment on protection against soybean rust and corn rust 1. Preparation of spore suspension: Fresh soybean rust leaves were thoroughly shaken and filtered in a sealed container with sterile water containing 0.1 wt% Tween 80; the urediniospores were counted under a microscope using a hemocytometer to prepare a soybean rust urediniospore suspension with a concentration of 5 × 10⁹ spores / mL.

[0067] 2. Determination of preventive efficacy: Soybean seedlings were cultivated in greenhouse incubators. When the fourth true leaf of the soybean plant was fully expanded, different concentrations of the pesticide (solvent: sterile water), 30wt% fluopyram, and sterile water were sprayed onto the soybean seedlings (both the upper and lower surfaces of the leaves needed to be covered with the pesticide solution). Ten soybean seedlings were sprayed for each treatment. After the pesticide solution on the leaves was air-dried, urediniospore suspension was inoculated on both the upper and lower surfaces of the leaves.

[0068] Test example: Experiment on treating soybean rust 1. Preparation of spore suspension: Fresh soybean rust leaves were thoroughly shaken and filtered in a sealed container with sterile water containing 0.1 wt% Tween 80; the urediniospores were counted under a microscope using a hemocytometer to prepare a soybean rust urediniospore suspension with a concentration of 5 × 10⁹ spores / mL.

[0069] 2. Culture of diseased plants: Inoculate soybean leaves with 2*10 (5-fold) spores / mL of soybean rust and place them in a humidifier for humidification. 24 hours after inoculation, remove the soybeans from the humidifier and air dry them. Use a throat sprayer to evenly spray the pesticide onto soybean leaves with two true leaves. Repeat each treatment three times. After spraying, place the plants indoors to air dry naturally, and then place them in a culture room for 13 days.

[0070] 3. Treatment efficacy determination: Different concentrations of the drug (solvent: sterile water), 30wt% fluoxetine, and sterile water were sprayed onto the diseased leaves (both the upper and lower surfaces of the leaves needed to be covered with the solution). Each treatment was applied to the diseased leaves of 10 soybean seedlings. The experiment was repeated 3 times, the disease index was recorded, and the treatment efficacy was calculated (the percentage of diseased leaf area was used to evaluate the fungal invasion on the leaves).

[0071] Test Example: Experiment on Protection Against Corn Rust 1. Preparation of spore suspension: Fresh corn rust leaves were thoroughly mixed with sterile water containing 0.1 wt% Tween 80 in a sealed container and filtered; the urediniospores were counted under a microscope using a hemocytometer to prepare a corn rust urediniospore suspension with a concentration of 5 × 10⁹ spores / mL.

[0072] 2. Determination of preventive efficacy: Maize seedlings were cultivated in greenhouse incubators. When the fourth true leaf of the maize seedling was fully expanded, different concentrations of the pesticide, 30wt% fluopyram, and sterile water were sprayed onto the seedlings (both the upper and lower surfaces of the leaves needed to be covered with the pesticide solution). Ten maize seedlings were sprayed for each treatment. After the pesticide solution on the leaves was air-dried, urediniospore suspension was inoculated on both the upper and lower surfaces of the leaves.

[0073] Test Example: Experiment on Treatment of Corn Rust 1. Preparation of spore suspension: Fresh corn rust leaves were thoroughly mixed with sterile water containing 0.1 wt% Tween 80 in a sealed container and filtered; the urediniospores were counted under a microscope using a hemocytometer to prepare a corn rust urediniospore suspension with a concentration of 5 × 10⁹ spores / mL.

[0074] 2. Culture of diseased plants: Inoculate soybean leaves with 2*10 (5-fold) spores / mL of corn rust and place them in a humidifier for humidification. 24 hours after inoculation, remove the corn plants from the humidifier and air dry them. Use a throat sprayer to evenly spray the fungicide onto the two true leaf stages of the corn leaves. Repeat each treatment three times. After spraying, place the plants indoors to air dry naturally, and then place them in a culture room for 13 days.

[0075] 3. Treatment efficacy determination: Different concentrations of the drug, 30wt% fluopyram, and sterile water were sprayed onto the diseased leaves (both the upper and lower surfaces of the leaves needed to be covered with the solution). Each treatment was applied to the diseased leaves of 10 maize seedlings. The experiment was repeated 3 times, and the disease index was recorded to calculate the treatment efficacy (the percentage of diseased leaf area was used to evaluate the fungal invasion on the leaves).

[0076] Disease index = × 100 Prevention effect (%) = × 100% Table 2 Grading Standards for Soybean Rust and Corn Rust 3. Experimental Results: See Tables 3 and 4 for details.

[0077] Table 3. Experimental results of protective effects against soybean rust and corn rust. Table 4. Experimental results of treatment effects on soybean rust and corn rust. As shown in Tables 3 and 4, the compounds of this invention exhibit good inhibitory effects on soybean rust spore germination, and also demonstrate good protective and curative activity against soybean rust. Several lead compounds showed inhibition rates exceeding 77% at a dosage concentration of 5 mg / L, comparable to the control agent fluoxetine.

[0078] As shown in Tables 3 and 4, the compounds of this invention exhibit good inhibitory effects on the germination of corn rust spores, and also demonstrate good protective and therapeutic activity against corn rust. Several lead compounds showed inhibition rates exceeding 77% at a dosage concentration of 5 mg / L, comparable to the control agent fluoxetine.

[0079] The standard C and H spectra of compound 24 are as follows: Figures 1-2 The specific data is as follows: 1 H NMR (400 MHz, CDCl3) δ 8.16 (d, J = 8.4 Hz, 2H), 7.80 (d, J = 8.4Hz, 2H), 4.35 (s, 2H), 4.28 (s, 2H), 0.75 – 0.65 (m, 4H) ppm. 13 C NMR (101 MHz, CDCl3) δ 168.82, 168.66, 166.15 (q, 2 J = 44.4 Hz),137.14, 128.77, 127.86, 127.13, 116.03 (q, 1 J = 274.7 Hz), 61.17, 56.85,15.62, 10.03 ppm. Molecular docking technology can elucidate the binding modes of compounds to target proteins. A three-dimensional structural model of *Aspergillus nidus* HdaA was constructed using AlphaFold3. To predict the binding mode, Flufenoxadiazam and A7 were molecularly docked with the AlphaFold3-predicted *Aspergillus nidus* HdaA model and human HDAC4, respectively. The docking results are shown below. Figure 3 As shown.

[0080] The 5-(trifluoromethyl)-1,2,4-oxadiazole fragments of compounds 24 and 57 are deeply embedded within the HdaA active pocket. At the bottom of this pocket, the oxygen atom on the oxadiazole ring and a fluorine atom in the CF3 group directly react with the catalytic Zn. 2+ The ions coordinate at distances of 2.6 Å and 3.2 Å, respectively. Simultaneously, the oxadiazole ring forms significant π-π interactions with His-268 and His-309 residues, while the benzene ring in its benzoic acid structure exhibits π-π interactions with the Phe-227 residue.

[0081] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. A compound containing trifluoromethyloxadiazole and a spirocyclic structure, or a pesticide-acceptable salt, hydrate, or solvate thereof, characterized in that, The structure of the compound containing trifluoromethyloxadiazole and a spirocyclic structure is shown in general formula I: General Formula I Wherein, X is any one of ester group, carbonyl group, amide group or oxime ester group; M can be either a nitrogen atom or a carbon atom; Ring A is an unsubstituted saturated 4- to 6-membered ring; Ring B is a saturated 3- to 6-membered ring containing 0 to 2 heteroatoms, or a benzo[a]-fused ring containing 0 to 1 oxygen atom; R1 is located at The hydrogen, C1-C4 alkyl, C1-C4 alkoxy, cyano or halogen at any substitution position on the ring; R2 and R3 are each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, nitro, cyano, carbonyl, amino, methoxy, ethoxy, propylamino, isooxypropyl, n-butoxy, tert-butoxy, cyclohexyloxy, N-(N,N-dimethyl) substituent, N-(N,N-diethyl) substituent, N-(N,N-dipropyl) substituent, N-(N,N-dipropyl) substituent, N-(N, N-Dibutyl) substituent, N-(N-methyl-N-ethyl) substituent, N-(N-methyl-N-propyl) substituent, N-(N-methyl-N-butyl) substituent, N-(N-ethyl-N-propyl) substituent, N-(N-ethyl-N-butyl) substituent, N-(N-propyl-N-butyl) substituent, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 2 or 3 or 4-methylcyclohexyl, 2 or 3 or 4-ethylcyclohexyl, 2 or 3 or 4-hydroxycyclohexyl, 2 or 3 or 4-carboxycyclohexyl, 2 or 3 or 4-cyanocyclohexyl, 2 or 3 or 4-aminocyclohexyl, 2 or 3-tetrahydrofuranyl, 2 or 3-tetrahydrothiophenyl, N or 2 Or any one of 3-tetrahydropyrrole, 2, 3, or 4-tetrahydropyranyl, N, 2, 3, or 4-piperidinyl, N, 2, or 3-morpholinyl, N, 2-piperazinyl, N, 2, 3, or 3-(N-methylpiperazinyl), N, 2, 3, or 3-(N-benzylpiperazinyl)phenyl, 2, 3, or 4-methylphenyl, 2, 3, or 4-methoxyphenyl, 2, 3, or 4-aminophenyl, 2, 3, or 4-cyanophenyl, 2, 3, or 4-carboxyphenyl, 2, 3, or 4-nitrophenyl, 2, 3, or 4-hydroxyphenyl, naphthyl, 2, 3-furanyl, 2, 3-thienyl, N, 2, 3, or 3-pyrrole, 2, 3, or 4-pyridinyl, N, 2, or 3-(N-morpholinyl)phenyl, and indoleyl.

2. The compound containing trifluoromethyloxadiazole and a spirocyclic structure, or its pesticide-acceptable salt, hydrate, or solvate, as described in claim 1, is characterized in that... The structure of the compound containing trifluoromethyloxadiazole and a spirocyclic structure is shown in general formula II: General Formula II Where M is a nitrogen atom; X can be either a carbonyl group or an amide group; Ring A is an unsubstituted saturated 4- to 6-membered ring; Ring B is a substituted or unsubstituted saturated 3- to 6-membered ring containing 0 to 2 heteroatoms, or a benzo[a] heterocycle containing 0 to 1 oxygen atom; R1 is hydrogen; R2 and R3 are each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, nitro, cyano, carbonyl, amino, methoxy, ethoxy, propylamino, isooxypropyl, n-butoxy, tert-butoxy, cyclohexyloxy, N-(N,N-dimethyl) substituent, N-(N,N-diethyl) substituent, N-(N,N-dipropyl) substituent, N-(N,N-dipropyl) substituent, N-(N, N-Dibutyl) substituent, N-(N-methyl-N-ethyl) substituent, N-(N-methyl-N-propyl) substituent, N-(N-methyl-N-butyl) substituent, N-(N-ethyl-N-propyl) substituent, N-(N-ethyl-N-butyl) substituent, N-(N-propyl-N-butyl) substituent, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 2 or 3 or 4-methylcyclohexyl, 2 or 3 or 4-ethylcyclohexyl, 2 or 3 or 4-hydroxycyclohexyl, 2 or 3 or 4-carboxycyclohexyl, 2 or 3 or 4-cyanocyclohexyl, 2 or 3 or 4-aminocyclohexyl, 2 or 3-tetrahydrofuranyl, 2 or 3-tetrahydrothiophenyl, N or 2 Or any one of 3-tetrahydropyrrole, 2, 3, or 4-tetrahydropyranyl, N, 2, 3, or 4-piperidinyl, N, 2, or 3-morpholinyl, N, 2-piperazinyl, N, 2, 3, or 3-(N-methylpiperazinyl), N, 2, 3, or 3-(N-benzylpiperazinyl)phenyl, 2, 3, or 4-methylphenyl, 2, 3, or 4-methoxyphenyl, 2, 3, or 4-aminophenyl, 2, 3, or 4-cyanophenyl, 2, 3, or 4-carboxyphenyl, 2, 3, or 4-nitrophenyl, 2, 3, or 4-hydroxyphenyl, naphthyl, 2, 3-furanyl, 2, 3-thienyl, N, 2, 3, or 3-pyrrole, 2, 3, or 4-pyridinyl, N, 2, or 3-(N-morpholinyl)phenyl, and indoleyl.

3. A compound containing trifluoromethyloxadiazole and a spirocyclic structure, or a pesticide-acceptable salt, hydrate, or solvate thereof, as described in claim 2, is characterized in that... X is a carbonyl group; ring A is an unsubstituted saturated 4- to 5-membered ring; ring B is an unsubstituted saturated 3- to 5-membered ring or a benzo[a]heterocyclic ring containing 0 to 1 oxygen atom.

4. A compound containing trifluoromethyloxadiazole and a spirocyclic structure, or a pesticide-acceptable salt, hydrate, or solvate thereof, as described in claim 1, characterized in that... The structure of the compound containing trifluoromethyloxadiazole and a spirocyclic structure is shown in general formula III: Formula III Where M is a carbon atom; X is any one of ester, carbonyl, amide, or oxime ester; Ring A is an unsubstituted saturated 4- to 6-membered ring; Ring B is a substituted or unsubstituted saturated 3- to 6-membered ring containing 0 to 2 heteroatoms, or a benzo[a] heterocycle containing 0 to 1 oxygen atom; R1 is located at The hydrogen, C1-C4 alkyl, C1-C4 alkoxy, cyano or halogen at any substitution position on the ring; R2 and R3 are each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, nitro, cyano, carbonyl, amino, methoxy, ethoxy, propylamino, isooxypropyl, n-butoxy, tert-butoxy, cyclohexyloxy, N-(N,N-dimethyl) substituent, N-(N,N-diethyl) substituent, N-(N,N-dipropyl) substituent, N-(N,N-dipropyl) substituent, N-(N, N-Dibutyl) substituent, N-(N-methyl-N-ethyl) substituent, N-(N-methyl-N-propyl) substituent, N-(N-methyl-N-butyl) substituent, N-(N-ethyl-N-propyl) substituent, N-(N-ethyl-N-butyl) substituent, N-(N-propyl-N-butyl) substituent, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 2 or 3 or 4-methylcyclohexyl, 2 or 3 or 4-ethylcyclohexyl, 2 or 3 or 4-hydroxycyclohexyl, 2 or 3 or 4-carboxycyclohexyl, 2 or 3 or 4-cyanocyclohexyl, 2 or 3 or 4-aminocyclohexyl, 2 or 3-tetrahydrofuranyl, 2 or 3-tetrahydrothiophenyl, N or 2 Or any one of 3-tetrahydropyrrole, 2, 3, or 4-tetrahydropyranyl, N, 2, 3, or 4-piperidinyl, N, 2, or 3-morpholinyl, N, 2-piperazinyl, N, 2, 3, or 3-(N-methylpiperazinyl), N, 2, 3, or 3-(N-benzylpiperazinyl)phenyl, 2, 3, or 4-methylphenyl, 2, 3, or 4-methoxyphenyl, 2, 3, or 4-aminophenyl, 2, 3, or 4-cyanophenyl, 2, 3, or 4-carboxyphenyl, 2, 3, or 4-nitrophenyl, 2, 3, or 4-hydroxyphenyl, naphthyl, 2, 3-furanyl, 2, 3-thienyl, N, 2, 3, or 3-pyrrole, 2, 3, or 4-pyridinyl, N, 2, or 3-(N-morpholinyl)phenyl, and indoleyl.

5. A compound containing trifluoromethyloxadiazole and a spirocyclic structure, or a pesticide-acceptable salt, hydrate, or solvate thereof, as described in claim 4, is characterized in that... X is a carbonyl group; ring A is an unsubstituted saturated 4- to 5-membered ring; ring B is an unsubstituted saturated 3- to 5-membered ring or a benzo[a]heterocyclic ring containing 0 to 1 oxygen atom; R1 is hydrogen.

6. A compound containing trifluoromethyloxadiazole and a spirocyclic structure, or a pesticide-acceptable salt, hydrate, or solvate thereof, as described in claim 1, characterized in that... The structural formula of the compound containing trifluoromethyloxadiazole and a spirocyclic structure is selected from any of the following:

7. The use of a compound containing trifluoromethyloxadiazole and a spirocyclic structure as described in any one of claims 1 to 7, or a pesticide-acceptable salt, hydrate, or solvate thereof, in the preparation of histone deacetylase inhibitors.

8. The use of a compound containing trifluoromethyloxadiazole and a spirocyclic structure as described in any one of claims 1 to 7, or its phytochemically acceptable salt, hydrate, or solvate, in the control of plant fungal diseases.

9. The application according to claim 8, characterized in that, The plant fungal diseases mentioned are diseases caused by any one or more of the following pathogens: soybean rust, corn rust, wheat leaf rust, wheat stripe rust, tomato gray mold, rapeseed sclerotinia, tomato early blight, and wheat stem rust.

10. A pharmaceutical composition, characterized in that, It includes a therapeutically effective amount of an active ingredient and a pharmaceutically and pesticide-acceptable carrier, wherein the active ingredient is at least one of the following: a compound containing a trifluoromethyloxadiazole and a spirocyclic structure as described in any one of claims 1 to 7, a pharmaceutically and pesticide-acceptable salt thereof, or a pharmaceutically and pesticide-acceptable solvate thereof.

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