Pyran compounds as plant immune activators and uses thereof

Abstract

The invention provides a 2-amino-3-cyano-4-aryl-4H-pyran compound as shown in a formula I in the specification, or an optical isomer, a cis-trans-isomer or a pharmaceutically acceptable salt of the 2-amino-3-cyano-4-aryl-4H-pyran compound. The compound provided by the invention has good plant immune activation activity, so that the compound can be used as a plant immune activator. The invention also provides a pesticide composition containing the compound. The compound and the pesticide composition have the advantages of being environmentally friendly, safe, free of drug resistance and the like, and therefore the compound and the pesticide composition have good application prospects. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of pesticides, in particular to pyran compounds, and the use of these compounds as plant immune activators and methods for controlling insect pests in agricultural, forestry and horticultural environments using these compounds or compositions containing them. Background Art

[0002] Pesticides refer to drugs used to prevent and control various diseases and regulate crop growth in agricultural production, which can significantly improve the quality of crops and increase yields. Most traditional pesticides are aimed at directly killing harmful organisms. They often achieve good results in the early stages of use. However, if they are used in large quantities for a long time, many problems will arise, including environmental pollution, increased drug resistance, and adverse effects such as pesticide residues. Food is the primary need of the people. As living standards continue to improve, the requirements for food quality are becoming more stringent. Chemical fungicides have been used to prevent and control plant diseases for hundreds of years, from the initial inorganic compound copper sulfate and highly toxic organic metal compounds to today's new high-efficiency and low-toxic fungicides.

[0003] Under natural growth conditions, plants are constantly faced with various stresses and competitions, including physical and chemical stimuli such as ultraviolet rays and drought, as well as infections from pathogenic microorganisms and pests. Therefore, after a long period of evolution, plants have developed a variety of resistance mechanisms and immune systems to resist adverse external factors and alleviate damage symptoms. In addition to physical barriers such as epidermal cells and guard cells, plants have also evolved an immune defense response induced by recognizing special pathogen-related molecules, including systemic acquired resistance (SAR) and induced systemic resistance (ISR). Although there are no specific immune cells in plants, there are two major types of immune receptors on each cell that recognize pathogens, including pattern recognition receptors (PRRs) located on the cell membrane surface that can recognize pathogen-associated molecular patterns (PAMPs), and nucleotide binding and leucine-rich repeat receptors (NLRs) located in the cell, which are used to mediate plant immune responses. After PRRs recognize conserved PAMPs, they activate pattern-triggered immunity (PTI) to resist disease infection through reactive oxygen bursts and secretion of a variety of plant endogenous hormones. Since these components are ubiquitous in plant cells, they do not have specific immunity and mainly work in the early stages of plant diseases. However, as a countermeasure, pathogens secrete effector molecules to inhibit the PTI response and infect plants. This process is called effector-triggered susceptibility. At the same time, in order to prevent further infection by pathogens, plants have evolved NLRs proteins encoded by intracellular disease-resistant genes, which can recognize and activate effector-triggered immunity (ETI) and produce a stronger defense response as a second layer of the immune system.

[0004] Plant immune activators are different from traditional pesticides. They have no in vitro bactericidal activity. They can activate the plant immune system to produce disease resistance under living conditions, thereby preventing plants from being invaded by pathogens. The mechanism of action of plant immune activators is relatively complex. They do not exert their efficacy on a single target, but induce the plant system to form overall disease resistance. Therefore, they are safer than traditional pesticides. They can not only avoid the development of drug resistance due to long-term use, but also effectively reduce the adverse effects on the environment and non-target organisms. Because they play an immune activation role, they need to be used for a certain period before plant diseases occur. Plant immune activators are widely used and can produce protective effects on a variety of plants and pathogens. They can also be used in combination with other pesticides to enhance the effect. Although a variety of plant immune activators have been commercialized, they are still significantly insufficient compared to the huge market demand; their mechanism of action is relatively complex and there is no clear target, and research in this field is far from in-depth. Therefore, it is very necessary to develop new plant immune activators and study the disease resistance mechanism. Summary of the invention

[0005] The purpose of the present invention is to provide a pyran compound with a novel structure, which is a new type of plant immune activator, thereby activating the immune system of plants to enhance the ability to resist external invasion, thereby achieving the purpose of preventing and controlling plant diseases.

[0006] Another object of the present invention is to provide an agricultural composition comprising the pyran compound, and the use of the pyran compound or the agricultural composition for controlling pests in agricultural, forestry and horticultural environments.

[0007] Another object of the present invention is to provide a method for controlling pests in agricultural, forestry and horticultural environments using the pyran compounds or agricultural compositions.

[0008] In the first aspect, the present invention provides a compound of formula I, or an optical isomer, a cis-trans isomer or a pharmaceutically acceptable salt thereof,

[0009]

[0010] wherein R1 is selected from the group consisting of substituted or unsubstituted C5-C8 aryl (preferably phenyl), substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 epoxyalkyl, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 alkylthio, substituted or unsubstituted C1-C6 alkylsulfonyl, substituted or unsubstituted C l -C6 alkyl acyl, substituted or unsubstituted C l-C6 alkyl ester group, substituted or unsubstituted C l -C6 alkylamide;

[0011] R2 is selected from the following group: substituted or unsubstituted C5-C8 aryl (preferably phenyl), substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 epoxyalkyl, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 alkylthio, substituted or unsubstituted C1-C6 alkylsulfonyl, substituted or unsubstituted C l -C6 alkyl acyl, substituted or unsubstituted C l -C6 alkyl ester group, substituted or unsubstituted C l -C6 alkylamide;

[0012] Ar is selected from the following group: a substituted or unsubstituted monocyclic, bicyclic or tricyclic aromatic group containing 6 to 14 carbon atoms, or a substituted or unsubstituted 5 to 10 membered monocyclic, bicyclic or tricyclic heteroaromatic group containing 1 to 3 heteroatoms selected from N, O or S.

[0013] In a specific embodiment, R1 is selected from the group consisting of substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 cycloalkyl;

[0014] R2 is selected from the group consisting of substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy;

[0015] Ar is selected from the group consisting of substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, or substituted or unsubstituted 5-10 membered monocyclic or bicyclic heteroaryl.

[0016] In a specific embodiment, R1 is a substituted or unsubstituted C1-C4 alkyl group;

[0017] R2 is a substituted or unsubstituted C1-C4 alkyl group;

[0018] Ar is a substituted or unsubstituted phenyl group, or a substituted or unsubstituted 5-10 membered heteroaryl group.

[0019] In a preferred embodiment, R1 and R2 are independently substituted by halogen.

[0020] In a preferred embodiment, Ar is optionally substituted with 0-4 substituents selected from the group consisting of halogen, hydroxy, amino, nitro, cyano, phenyl, optionally halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, optionally halogenated C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 epoxyalkyl, C1-C6 alkylamino, C1-C6 alkylthio, C1-C6 alkylsulfonyl, C l -C6 alkyl acyl, C l -C6 alkyl ester group, C l -C6 alkylamide.

[0021] In a preferred embodiment, Ar is optionally substituted by 0-3 substituents selected from the group consisting of halogen, nitro, cyano, optionally halogenated C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkylthio.

[0022] In a specific embodiment, Ar is phenyl, furanyl, thienyl, thiazolyl, isoxazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl or isothiazolyl, and is optionally substituted with 1-3 substituents selected from the group consisting of halogen, nitro, cyano, methyl, methoxy, methylthio, trifluoromethyl;

[0023] Preferably, Ar is phenyl, thiazolyl or isothiazolyl, and is optionally substituted by 1-2 substituents selected from the group consisting of halogen, methyl, trifluoromethyl;

[0024] Preferably, Ar is thiazolyl or isothiazolyl, and is optionally substituted by 1-2 substituents selected from the group consisting of halogen, methyl, trifluoromethyl.

[0025] In a specific embodiment, the compound is a compound selected from the group consisting of:

[0026]

[0027]

[0028]

[0029] In a specific embodiment, the compound is selected from the group consisting of:

[0030]

[0031] Preferably, the compound is selected from the group consisting of:

[0032]

[0033] In a second aspect, the present invention provides a pesticide composition, characterized in that the pesticide composition contains:

[0034] (1) 0.001-99.99 wt% of the compound of the first aspect, or its optical isomers, cis-trans isomers, or pesticide-acceptable salts thereof, in an amount of 0.001-99.99 wt% of the total weight of the pharmaceutical composition; and

[0035] (2) A pesticide-acceptable carrier or excipient.

[0036] In a preferred embodiment, the pesticide composition is a solution, emulsion, suspension, powder, foam, paste, granule; aerosol, natural and synthetic materials impregnated with active substances, microcapsules in polymers, coating compounds for seeds, formulations for use with combustion devices (such as fumigation cartridges, fumigation pots and fumigation trays), ULV cold mist or hot mist formulations; or

[0037] The pesticide composition further comprises other active compounds, which are selected from the group consisting of insecticides, baits, bactericides, miticides, nematicides, fungicides, and growth control agents.

[0038] In a preferred embodiment, the pesticide composition is a plant immune activator that activates the immune system of a plant to resist diseases.

[0039] In a preferred embodiment, the plant is an agricultural crop.

[0040] In a preferred embodiment, the agricultural crops include cucumber, rice, wheat, tomato, corn and the like.

[0041] In a preferred embodiment, the diseases include rice sheath blight, rice blast, wheat fusarium sclerotinia, rapeseed sclerotinia, cucumber powdery mildew, cucumber gray mold, cucumber fusarium sclerotium, cucumber vine blight, cucumber wilt, tomato infestation, corn leaf blight, etc.

[0042] In a third aspect, the present invention provides use of the compound described in the first aspect, or its optical isomers, cis-trans isomers or pesticide-acceptable salts in the preparation of pesticides.

[0043] In a preferred embodiment, the pesticide is a plant immune activator that activates the plant's immune system to resist disease.

[0044] In a preferred embodiment, the plant is an agricultural crop.

[0045] In a preferred embodiment, the agricultural crops include cucumber, rice, wheat, tomato, corn and the like.

[0046] In a preferred embodiment, the diseases include rice sheath blight, rice blast, wheat fusarium sclerotinia, rapeseed sclerotinia, cucumber powdery mildew, cucumber gray mold, cucumber fusarium sclerotium, cucumber vine blight, cucumber wilt, tomato infestation, corn leaf blight, etc.

[0047] In a fourth aspect, the present invention provides a method for preventing and controlling plant diseases, characterized in that the method comprises: administering an effective amount of the compound described in the first aspect or the pesticide composition described in the second aspect to the plant, thereby preventing and controlling the corresponding disease.

[0048] In a preferred embodiment, the plant is an agricultural crop.

[0049] In a preferred embodiment, the agricultural crops include cucumber, rice, wheat, tomato, corn and the like.

[0050] In a preferred embodiment, the diseases include rice sheath blight, rice blast, wheat fusarium sclerotinia, rapeseed sclerotinia, cucumber powdery mildew, cucumber gray mold, cucumber fusarium sclerotium, cucumber vine blight, cucumber wilt, tomato infestation, corn leaf blight, etc.

[0051] In a fifth aspect, the present invention provides a method for preparing the compound of the first aspect, characterized in that the compound is prepared using the general synthetic route shown below:

[0052]

[0053] Wherein, R1, R2 and Ar are as described in the first aspect.

[0054] In a preferred embodiment, the preparation method comprises:

[0055] The first step is to use various substituted five-membered or six-membered aromatic aldehyde compounds (structure shown in formula A) as starting materials and malononitrile (structure shown in formula B) to undergo Knoevenagel condensation reaction in the presence of a basic catalyst. After the reaction is completed, the intermediate B is obtained by suction filtration and used for subsequent reactions without further purification.

[0056] The second step reaction is that the intermediate B and methyl acetoacetate undergo Michael addition and Thorpe cyclization rearrangement reaction in the presence of an alkaline catalyst. After the reaction is completed, the target product 2-amino-3-cyano-4-aryl-4H-pyran compound is obtained by filtration and recrystallization to obtain a pure product.

[0057] In a preferred embodiment, the solvents that can be used in the preparation method include: alcohols such as methanol, ethanol, isopropanol, etc.; chain or cyclic ethers such as diethyl ether, tetrahydrofuran, 1,4-dioxane, etc.; ketones such as acetone and butanone, etc.; aromatic hydrocarbons such as benzene and toluene, etc.; nitriles such as acetonitrile and propionitrile, etc.; amides such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, etc.; sulfones such as dimethyl sulfoxide and cyclopentane, etc.; water; or a mixture of two or more thereof.

[0058] In a preferred embodiment, the alkaline catalyst that can be used in the preparation method includes: inorganic bases such as ammonia, sodium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium fluoride, ammonium acetate, etc.; organic bases such as pyridine, piperidine, diethylamine, triethylamine, tetrahydropyrrole, etc.; or a mixture of two or more thereof.

[0059] In a preferred embodiment, the solvents that can be used for the recrystallization include: alcohols such as methanol, ethanol, isopropanol, etc.; linear or cyclic ethers such as diethyl ether, isopropyl ether, methyl tert-butyl ether, etc.; halogenated hydrocarbons such as dichloromethane and chloroform, etc.; esters such as ethyl acetate and isopropyl acetate, etc.; alkanes such as n-hexane, n-heptane, petroleum ether, etc.; or a mixture of two or more thereof.

[0060] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0061] After extensive and in-depth research, the inventors designed and synthesized a series of 2-amino-3-cyano-4-aryl-4H-pyran compounds. After biological activity tests on these compounds, it was found that some of them had good plant immune activation activity, thus having good research value and application prospects. On this basis, the present invention was completed.

[0062] definition

[0063] The terms used herein have the meanings commonly understood by those skilled in the art to which the present invention belongs. For the purpose of a clear understanding of the present invention, some of the terms used herein are defined as follows:

[0064] The term "alkyl" as used herein refers to straight and branched chain alkyl groups, usually 1-6 carbon atoms, preferably 1-4 carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, etc.

[0065] The term "alkoxy" as used herein refers to an "alkyl-O-" group, wherein the alkyl group may be a C1-C6 straight chain or branched chain alkyl group, preferably a C1-C4 straight chain or branched chain alkyl group.

[0066] The term "alkylthio" as used herein refers to an "alkyl-S-" group, wherein the alkyl group may be a C1-C6 straight chain or branched chain alkyl group, preferably a C1-C4 straight chain or branched chain alkyl group.

[0067] In the present invention, the alkyl group may be optionally substituted with halogen. Examples of halogenated alkyl groups include C1-C6 alkyl groups substituted with 1-3 F, Cl, or Br atoms, and specific examples include trifluoromethyl.

[0068] The term "aryl" as used herein refers to monocyclic, bicyclic and tricyclic aromatic groups containing 6-14 carbon atoms. Examples of aryl groups include phenyl, naphthyl, phenanthrenyl, anthracenyl, indenyl, azulenyl, fluorenyl, tetrahydronaphthyl, indanyl and the like.

[0069] In the present invention, the aryl group may be optionally substituted by 1-5 identical or different substituents selected from the following: halogen, amino, nitro, cyano, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, etc.

[0070] The term "heteroaryl" as used herein refers to a ring containing 5-10 atoms and 6-14 electrons shared in the ring system, wherein the atoms contained in the ring are carbon atoms and 1-3 heteroatoms selected from N, O, S which may be the same or different.

[0071] In the present invention, examples of heteroaryl include pyridyl, quinolyl, isoquinolyl, furanyl, thienyl, pyrrolyl, benzofuranyl, benzothienyl, indolyl, imidazolyl, pyrazolyl, thiazolyl, isoxazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl, isothiazolyl, benzothiazolyl, benzimidazolyl, etc. In the present invention, heteroaryl may be optionally substituted with 1-5 identical or different substituents selected from the following: halogen, amino, nitro, cyano, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, etc.

[0072] Compounds of the present invention and preparation methods and uses thereof

[0073] It is known in the art that 2-amino-3-cyano-4-aryl-4H-pyran compounds have a variety of biological activities, such as antibacterial, antitumor, anti-rheumatic, etc., and have been widely used in the medical field. The inventors have found that 2-amino-3-cyano-4-aryl-4H-pyran derivatives having the parent core structure also have good plant immune activation activity.

[0074] In this document, "compounds of the present invention" or "pyran compounds of the present invention" have the same meaning, and both refer to compounds of formula I, or optical isomers, cis-trans isomers or pharmaceutically acceptable salts thereof:

[0075]

[0076] In the formula, each substituent has the same meaning as defined above.

[0077] Based on the structure of the compound of the present invention, those skilled in the art know how to synthesize 2-amino-3-cyano-4-aryl-4H-pyran derivatives. However, it is preferred to use the method of the present invention to prepare the compound of formula I. The synthetic strategy of the preparation method of the present invention is as follows:

[0078]

[0079] In a specific embodiment, the general preparation method of the present invention comprises: the first step is to use various substituted five-membered or six-membered aromatic aldehyde compounds (structure shown in formula A) as starting materials, and malononitrile (structure shown in formula B) to undergo Knoevenagel condensation reaction in the presence of an alkaline catalyst, and after the reaction is completed, the intermediate B is filtered to obtain the intermediate B, which is used for subsequent reactions without further purification; the second step is to react the intermediate B with methyl acetoacetate in the presence of an alkaline catalyst to undergo Michael addition and Thorpe cyclization rearrangement reaction, and after the reaction is completed, the target product 2-amino-3-cyano-4-aryl-4H-pyran compound is filtered to obtain the target product, and the pure product is obtained by recrystallization. The solvents that can be used in the two-step reaction include: alcohols such as methanol, ethanol, isopropanol, etc.; chain or cyclic ethers such as diethyl ether, tetrahydrofuran, 1,4-dioxane, etc.; ketones such as acetone and butanone, etc.; aromatic hydrocarbons such as benzene and toluene, etc.; nitriles such as acetonitrile and propionitrile, etc.; amides such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, etc.; sulfones such as dimethyl sulfoxide and cyclopentane, etc.; water. These solvents can be used alone or in mixtures of two or more thereof. The basic catalysts that can be used in the two-step reaction include: inorganic bases such as ammonia, sodium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium fluoride, ammonium acetate, etc.; organic bases such as pyridine, piperidine, diethylamine, triethylamine, tetrahydropyrrole, etc.; these bases can be used alone or in mixtures of two or more thereof. Solvents that can be used for recrystallization include: alcohols such as methanol, ethanol, isopropanol, etc.; chain or cyclic ethers such as diethyl ether, isopropyl ether, methyl tert-butyl ether, etc.; halogenated hydrocarbons such as dichloromethane and chloroform, etc.; esters such as ethyl acetate and isopropyl acetate, etc.; alkanes such as n-hexane, n-heptane, petroleum ether, etc.; these solvents can be used alone or as a mixture of two or more thereof.

[0080] Since each reaction step involved in the preparation method is prior art, those skilled in the art can make appropriate selections and changes based on the synthesis strategies and post-processing steps described above.

[0081] Pesticide composition and method of use of the present invention

[0082] Since the compounds of the present invention can activate the immune system of plants to resist diseases, the present invention also provides a pesticide composition comprising the compounds of the present invention.

[0083] In a specific embodiment, the pesticide composition of the present invention comprises a compound represented by Formula I, or an optical isomer, a cis-trans isomer or a pharmaceutically acceptable salt thereof, and a pesticide-acceptable carrier.

[0084] The pesticide composition of the present invention may contain 0.01%wt-95%wt of the compound represented by formula I of the present invention as an active ingredient, or its optical isomers, cis-trans isomers or pharmaceutically acceptable salts.

[0085] The pesticide-acceptable carrier is known to those skilled in the art, including but not limited to solid carriers, liquid carriers, gaseous carriers, etc. In a specific embodiment, the solid carrier can be, for example: fine powders or granules of clay materials such as kaolin, diatomaceous earth, synthetic hydrated silica, bentonite, Fubasami clay and acid clay; fine powders or granules of various talcs, ceramics and other inorganic materials such as sericite, quartz, sulfur, activated carbon, calcium carbonate and hydrated silica; and fine powders or granules of fertilizers such as ammonium bicarbonate, ammonium sulfate, ammonium phosphate, ammonium nitrate, urea and ammonium chloride.

[0086] In a specific embodiment, the liquid carrier can be, for example: water; alcohols such as methanol, ethanol, n-propanol, isopropanol; linear or cyclic ethers such as diethyl ether, tetrahydrofuran, 1,4-dioxane, etc.; ketones such as acetone and butanone; aromatic hydrocarbons such as toluene; nitriles such as acetonitrile and propionitrile; amides such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone; sulfones such as dimethyl sulfoxide and cyclopentane sulfone, etc.; vegetable oils such as soybean oil and cottonseed oil.

[0087] The gas carrier or propellant may be, for example, Freon gas, butane gas, liquefied petroleum gas, dimethyl ether and carbon dioxide.

[0088] The pesticide composition of the present invention may further contain a surfactant such as alkyl sulfates, alkyl sulfonates, alkyl aryl sulfonates, alkyl aryl ethers and their polyethylene oxide derivatives, polyethylene glycol ethers, polyol esters and sugar alcohol derivatives.

[0089] The pesticide composition of the present invention may also contain adjuvants such as fixatives and dispersants, for example: casein, lignin derivatives, gelatin, polysaccharides (such as starch, gum arabic, cellulose derivatives and alginic acid), bentonite, and polymers such as polyvinyl alcohol, polyvinyl pyrrolidone, polyacrylic acid and other synthetic water-soluble polymers.

[0090] The pesticide composition of the present invention may also contain stabilizers, such as: PAP (isopropyl acid phosphate), BHT (2,6-di-tert-butyl-4-methylphenol), BHA (a mixture of 2-tert-butyl-4-methoxyphenol and 3-tert-butyl-4-methoxyphenol), vegetable oil, mineral oil, surfactant, fatty acid and its ester derivatives.

[0091] The pesticide composition of the present invention can be prepared by mixing the components of the pesticide composition of the present invention with each other.

[0092] The pesticide composition of the present invention thus prepared can be used directly or after dilution with water. In addition, it can also be used in admixture with other insecticides, acaricides, fungicides, antiviral agents, herbicides, plant growth regulators, synergists, fertilizers, soil conditioners and animal feeds or used simultaneously without admixture.

[0093] The pesticide composition of the present invention can activate the immune system of plants to resist diseases. In a specific embodiment, the plant is an agricultural crop. Further, the agricultural crops include cucumber, rice, wheat, tomato, corn, etc. In a specific embodiment, the diseases include rice sheath blight, rice blast, wheat fusarium, rape sclerotinia, cucumber powdery mildew, cucumber gray mold, cucumber fusarium, cucumber wilt, cucumber wilt, tomato late blight, corn leaf blight, etc.

[0094] Methods of controlling plant diseases

[0095] In view of the fact that the compound or pesticide composition of the present invention can activate the immune system of plants to resist diseases, the present invention also provides a method for controlling diseases, comprising administering an effective amount of the compound or pesticide composition of the present invention to plants, thereby controlling the corresponding diseases. In a specific embodiment, the method comprises spraying an effective amount of the compound or pesticide composition of the present invention on the surface of crop leaves, or applying it to the roots of crops in the soil, etc.

[0096] When the pesticide composition of the present invention is used in agriculture, the appropriate dosage and concentration can be set by those skilled in the art based on conditions including formulation type, frequency, location, and application method, disease type, and degree of damage.

[0097] Beneficial technical effects or advantages of the present invention:

[0098] 1. The compounds of the present invention are a series of plant immune activators with completely new structures;

[0099] 2. The compounds of the present invention are environmentally friendly;

[0100] 3. The compounds of the present invention are non-toxic or low-toxic to humans, beneficial insects or livestock, and thus have excellent safety; and

[0101] 4. The compounds of the present invention will not cause drug resistance.

[0102] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples that do not specify specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. The reagents used in the examples are all commercially available reagents. Unless otherwise stated, percentages and parts are calculated by weight.

[0103] Example 1. Preparation of methyl 2-amino-3-cyano-4-(4-trifluoromethylphenyl)-6-methyl-4H-pyran-5-carboxylate (A1)

[0104]

[0105] At room temperature, add 5 mL of anhydrous ethanol to a 50 mL eggplant-shaped flask, add 4-trifluoromethylbenzaldehyde (523 mg, 3.00 mmol, 1.0 eq.) and malononitrile (200 mg, 3.00 mmol, 1.0 eq.) to the reaction flask in sequence, stir to dissolve, and then add 1 drop of ammonia methanol solution (7 mol·L -1 ) as a catalyst. After 30 minutes, the reaction was complete by TLC detection. After filtration, the filter cake was washed twice with isopropyl ether to obtain 526 mg of 2-(4-trifluoromethylbenzylidene)-malononitrile as a yellow solid. The intermediate obtained did not need to be purified and was directly used for the next step. The crude product yield was 79%.

[0106] At room temperature, 5 mL of anhydrous ethanol was added to a 50 mL eggplant-shaped bottle, 2-(4-trifluoromethylbenzylidene)-malononitrile (526 mg, 2.37 mmol, 1.0 eq.) and methyl acetoacetate (300 mg, 2.60 mmol, 1.1 eq.) were added to the reaction bottle in sequence, and 1 drop of diethylamine was added as a catalyst. After 30 minutes, TLC showed that the reaction was complete. After suction filtration, the filter cake was washed twice with isopropyl ether, and then recrystallized twice with a mixed solvent of dichloromethane and petroleum ether to obtain the target compound A1, 562 mg of white solid. The yield was 70%, and the total yield was 56%.

[0107] Melting point: 159.8-161.3℃. 1 H NMR (400MHz, DMSO-d6) δ7.70(d,J=7.9Hz,2H),7.40(d,J=8.0Hz,2H),7.06(s,2H),4.43(s,1H),3.54(s,3H),2.36(s,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ166.3,159.1,158.2,150.0,128.4,127.8(q, 2 J C-F =31.8Hz),126.0(q,3 J C-F =4.0Hz),124.7(q, 1 J C-F =272.7Hz),119.9,106.8,57.0,52.1,39.2,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 16 H 13 F3N2O3:338.0878,Found338.0881.

[0108] Example 2. Preparation of methyl 2-amino-3-cyano-4-phenyl-6-methyl-4H-pyran-5-carboxylate (A2)

[0109]

[0110] Except that benzaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 51%.

[0111] Melting point: 176.0-178.0℃. 1 H NMR (400MHz, DMSO-d6) δ7.35-7.32(m,2H),7.25-7.22(m,1H),7.17-7.16(m,2H),6.93(s,2H),4.32(s,1H),3.54(s,3H),2.34(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.5,159.1,157.3,145.3,129.0,127.5,127.4,120.2,107.7,57.8,52.0,39.3,18.7ppm.HRMS(EI-TOF)Calculated[M] + for C 15 H 14 N2O3:270.1004,Found270.1003.

[0112] Example 3. Preparation of methyl 2-amino-3-cyano-4-(4-methylphenyl)-6-methyl-4H-pyran-5-carboxylate (A3)

[0113]

[0114] Except that p-tolualdehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 52%.

[0115] Melting point: 169.9-171.7℃. 1 H NMR (400MHz, DMSO-d6) δ7.11(d,J=7.8Hz,2H),7.01(d,J=7.7Hz,2H),6.88(s,2H),4.24(s,1H),3.52(s,3H),2.29(s,3H),2.26(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.6,159.0,157.0,142.4,136.4,129.6,127.5,120.2,107.8,57.9,52.0,38.9,21.1,18.7ppm.HRMS(EI-TOF)Calculated[M] + for C 16 H 16 N2O3:284.1161,Found284.1158.

[0116] Example 4. Preparation of methyl 2-amino-3-cyano-4-(4-bromophenyl)-6-methyl-4H-pyran-5-carboxylate (A4)

[0117]

[0118] Except that p-bromobenzaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 55%.

[0119] Melting point: 156.3-157.9℃. 1 H NMR (400MHz, DMSO-d6) δ7.53(d,J=8.5Hz,2H),7.14(d,J=8.4Hz,2H),7.00(s,2H),4.32(s,1H),3.55(s,3H),2.34(s,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ166.4,159.0,157.7,144.8,131.9,129.9,120.4,120.0,107.2,57.3,52.1,38.8,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 15 H 13 79 BrN2O3:348.0110,Found 348.0108.

[0120] Example 5. Preparation of methyl 2-amino-3-cyano-4-(4-chlorophenyl)-6-methyl-4H-pyran-5-carboxylate (A5)

[0121]

[0122] Except that p-chlorobenzaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 64%.

[0123] Melting point: 165.8-167.5℃. 1 H NMR (400MHz, DMSO-d6) δ7.39(d,J=8.0Hz,2H),7.19(d,J=8.0Hz,2H),6.99(s,2H),4.33(s,1H),3.54(s,3H),2.33(s,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ166.4,159.0,157.7,144.4,131.9,129.5,129.0,120.0,107.2,57.4,52.1,38.7,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 15 H 13 35 ClN2O3:304.0615,Found 304.0613.

[0124] Example 6. Preparation of methyl 2-amino-3-cyano-4-(4-fluorophenyl)-6-methyl-4H-pyran-5-carboxylate (A6)

[0125]

[0126] Except that p-fluorobenzaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 55%.

[0127] Melting point: 160.8-162.1℃. 1 H NMR (400MHz, DMSO-d6) δ7.22-7.13(m,4H),6.96(s,2H),4.34(s,1H),3.55(s,3H),2.33(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.4,161.6(d, 1 J C-F =243.4Hz),159.0,157.4,141.6,129.5(d,3 J C-F =8.0Hz),120.1,115.8(d, 2 J C-F =21.2Hz),107.6,57.6,52.0,38.6,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 15 H 13 FN2O3:288.0910,Found 288.0912.

[0128] Example 7. Preparation of methyl 2-amino-3-cyano-4-(4-methoxyphenyl)-6-methyl-4H-pyran-5-carboxylate (A7)

[0129]

[0130] Except that p-methoxybenzaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 53%.

[0131] Melting point: 153.6-155.3℃. 1 H NMR (400MHz, DMSO-d6) δ7.08(d,J=8.2Hz,2H),6.89(d,J=8.1Hz,2H),6.87(s,2H),4.27(s,1H),3.74(s,3H),3.55(s,3H),2.31(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.6,159.0,158.6,156.8,137.4,128.7,120.3,114.4,108.0,58.0,55.5,52.0,38.5,18.7ppm.HRMS(EI-TOF)Calculated[M] + for C 16 H 16 N2O4:300.1110,Found300.1112.

[0132] Example 8. Preparation of methyl 2-amino-3-cyano-4-(4-methylthiophenyl)-6-methyl-4H-pyran-5-carboxylate (A8)

[0133]

[0134] Except that p-trifluoromethylbenzaldehyde in Example 1 was replaced by p-methylthiobenzaldehyde, other conditions and steps were the same as those in Example 1. White solid, total yield 51%.

[0135] Melting point: 133.6-135.5℃. 1 H NMR (400MHz, DMSO-d6) δ7.22(d,J=8.2Hz,2H),7.10(d,J=8.3Hz,2H),6.94(s,2H),4.28(s,1H),3.55(s,3H),2.47(s,3H),2.32(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.5,159.0,157.3,142.1,137.0,128.2,126.6,120.2,107.6,57.6,52.1,38.8,18.8,15.2ppm.HRMS(EI-TOF)Calculated[M] + for C 16 H 16 N2O3S:316.0882,Found316.0880.

[0136] Example 9. Preparation of methyl 2-amino-3-cyano-4-(4-nitrophenyl)-6-methyl-4H-pyran-5-carboxylate (A9)

[0137]

[0138] Except that p-nitrobenzaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. Light yellow solid, total yield 60%.

[0139] Melting point: 170.3-171.9℃. 1 H NMR (400MHz, DMSO-d6) δ8.21(d,J=8.5Hz,2H),7.46(d,J=8.4Hz,2H),7.12(s,2H),4.50(s,1H),3.54(s,3H),2.37(s,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ166.2,159.1,158.6,153.0,146.9,128.9,124.4,119.8,106.5,56.7,52.2,39.2,18.9ppm.HRMS(EI-TOF)Calculated[M] + for C 15 H 13 N3O5:315.0855,Found 315.0852.

[0140] Example 10. Preparation of methyl 2-amino-3-cyano-4-(4-cyanophenyl)-6-methyl-4H-pyran-5-carboxylate (A10)

[0141]

[0142] Except that p-cyanobenzaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 65%.

[0143] Melting point: 193.9-195.8℃. 1 H NMR (400MHz, DMSO-d6) δ7.81(d,J=8.2Hz,2H),7.38(d,J=8.2Hz,2H),7.08(s,2H),4.43(s,1H),3.53(s,3H),2.36(s,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ166.2,159.1,158.5,150.9,133.1,128.7,119.9,119.3,110.2,106.6,56.8,52.1,39.4,18.9ppm.HRMS(EI-TOF)Calculated[M] + for C 16 H 13 N3O3:295.0957,Found 295.0960.

[0144] Example 11. Preparation of methyl 2-amino-3-cyano-4-(2-furyl)-6-methyl-4H-pyran-5-carboxylate (A11)

[0145]

[0146] Except that furan-2-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 50%.

[0147] Melting point: 214.0-216.0℃. 1 H NMR (400MHz, DMSO-d6) δ7.51(d,J=1.6Hz,1H),7.02(s,2H),6.35-6.33(m,1H),6.08(d,J=3.3Hz,1H),4.44(s,1H),3.62(s,3H),2.29(s,3H)ppm. 13C NMR(101MHz,DMSO-d6)δ166.3,160.1,158.5,156.4,142.7,120.1,111.0,105.8,105.5,54.6,52.2,32.9,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 12 N2O4:260.0797,Found 260.0794.

[0148] Example 12. Preparation of methyl 2-amino-3-cyano-4-(2-thienyl)-6-methyl-4H-pyran-5-carboxylate (A12)

[0149]

[0150] Except that thiophene-2-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 55%.

[0151] Melting point: 171.4-172.4℃. 1 H NMR (400MHz, DMSO-d6) δ7.36-7.34(m,1H),7.05(s,2H),6.94-6.92(m,1H),6.85-6.84(m,1H),4.65(s,1H),3.63(s,3H),2.28(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.3,159.6,157.4,149.8,127.5,125.3,124.4,120.1,108.1,57.4,52.2,34.3,18.7ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 12 N2O3S:276.0569,Found276.0567.

[0152] Example 13. Preparation of methyl 2-amino-3-cyano-4-(5-methyl-2-thienyl)-6-methyl-4H-pyran-5-carboxylate (A13)

[0153]

[0154] Except that 5-methyl-thiophene-2-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 53%.

[0155] Melting point: 156.7-158.6℃. 1 H NMR (400MHz, DMSO-d6) δ7.04(s,2H),6.64-6.60(m,2H),4.56(s,1H),3.66(s,3H),2.38(s,3H),2.29(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.3,159.6,157.2,147.4,138.6,125.5,124.1,120.1,108.1,57.4,52.2,34.5,18.7,15.5ppm.HRMS(EI-TOF)Calculated[M] + for C 14 H 14 N2O3S:290.0725,Found 290.0723.

[0156] Example 14. Preparation of methyl 2-amino-3-cyano-4-(3-thienyl)-6-methyl-4H-pyran-5-carboxylate (A14)

[0157]

[0158] Except that thiophene-3-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 55%.

[0159] Melting point: 180.0-181.7℃. 1 H NMR (400MHz, DMSO-d6) δ7.48-7.46(m,1H),7.19-7.18(m,1H),6.96(s,2H),6.93-6.91(m,1H),4.46(s,1H),3.62(s,3H),2.31(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.6,159.5,157.3,146.1,127.2,127.1,121.6,120.4,107.8,57.1,52.1,34.3,18.7ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 12 N2O3S:276.0569,Found276.0572.

[0160] Example 15. Preparation of methyl 2-amino-3-cyano-4-(5-nitro-2-thienyl)-6-methyl-4H-pyran-5-carboxylate (A15)

[0161]

[0162] Except that 5-nitro-thiophene-2-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. Light yellow solid, total yield 60%.

[0163] Melting point: 172.8-173.6℃. 1 H NMR (400MHz, DMSO-d6) δ8.00(d,J=4.3Hz,1H),7.33(s,2H),7.05(d,J=4.3Hz,1H),4.74(s,1H),3.68(s,3H),2.35(s,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ165.8,159.9,159.4,159.4,149.6,130.8,125.2,119.5,106.5,56.0,52.4,35.2,19.0ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 11 N3O5S:321.0419,Found 321.0416.

[0164] Example 16. Preparation of methyl 2-amino-3-cyano-4-(5-methyl-2-furyl)-6-methyl-4H-pyran-5-carboxylate (A16)

[0165]

[0166] Except that 5-methyl-furan-2-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 56%.

[0167] Melting point: 148.1-149.3℃. 1 H NMR (400MHz, DMSO-d6) δ7.00(s,2H),5.95(m,2H),4.39(s,1H),3.64(s,3H),2.30(s,3H),2.21(s,3H)ppm. 13C NMR(101MHz,DMSO-d6)δ166.4,160.1,158.2,154.6,151.2,120.1,107.0,106.5,105.6,54.8,52.2,33.0,18.7,13.9ppm.HRMS(EI-TOF)Calculated[M] + for C 14 H 14 N2O4:275.0954,Found 274.0951.

[0168] Example 17. Preparation of methyl 2-amino-3-cyano-4-(5-chloro-2-thienyl)-6-methyl-4H-pyran-5-carboxylate (A17)

[0169]

[0170] Except that 5-chloro-thiophene-2-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 63%.

[0171] Melting point: 167.1-168.8℃. 1 H NMR (400MHz, DMSO-d6) δ7.16(s,2H),6.94(d,J=3.8Hz,1H),6.74(d,J=3.9Hz,1H),4.59(s,1H),3.67(s,3H),2.30(s,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ166.1,159.7,158.0,149.1,127.2,124.4,119.8,110.4,107.4,56.6,52.3,34.8,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 11 35 ClN2O3S:310.0179,Found310.0177.

[0172] Example 18. Preparation of methyl 2-amino-3-cyano-4-(5-bromo-2-thienyl)-6-methyl-4H-pyran-5-carboxylate (A18)

[0173]

[0174] Except that 5-bromo-thiophene-2-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 62%.

[0175] Melting point: 161.0-162.2℃. 1 H NMR (400MHz, DMSO-d6) δ7.17(s,2H),7.06(d,J=3.8Hz,1H),6.72(d,J=3.8Hz,1H),4.62(s,1H),3.68(s,3H),2.30(s,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ166.1,159.7,158.1,151.7,130.8,125.4,119.9,110.3,107.4,56.6,52.3,34.7,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 11 79 BrN2O3S:353.9674,Found353.9676.

[0176] Example 19. Preparation of methyl 2-amino-3-cyano-4-(5-bromo-2-furyl)-6-methyl-4H-pyran-5-carboxylate (A19)

[0177]

[0178] Except that 5-bromo-furan-2-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 62%.

[0179] Melting point: 161.8-162.9℃. 1 H NMR (400MHz, DMSO-d6) δ7.01(s,2H),6.36(d,J=3.4Hz,1H),6.09(d,J=3.6Hz,1H),4.35(s,1H),3.55(s,3H),2.21(s,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ166.1,160.1,158.9,158.7,120.4,119.8,113.0,108.9,104.8,54.1,52.3,33.2,18.9ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 11 79 BrN2O4:337.9902,Found337.8999.

[0180] Example 20. Preparation of methyl 2-amino-3-cyano-4-(5-chloro-2-thienyl)-6-methyl-4H-pyran-5-carboxylate (A20)

[0181]

[0182] Except that 5-chloro-furan-2-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 64%.

[0183] Melting point: 155.4-156.4℃. 1 H NMR (400MHz, DMSO-d6) δ7.12(s,2H),6.36(d,J=3.2Hz,1H),6.22(d,J=3.5Hz,1H),4.44(s,1H),3.65(s,3H),2.32(s,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ166.1,160.1,158.9,156.5,134.0,119.8,108.6,108.2,104.8,54.1,52.3,33.2,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 11 35 ClN2O4:294.0407,Found294.0404.

[0184] Example 21. Preparation of methyl 2-amino-3-cyano-4-(5-cyano-2-thienyl)-6-methyl-4H-pyran-5-carboxylate (A21)

[0185]

[0186] Except that 5-cyano-thiazole-2-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 65%.

[0187] Melting point: 216.1-218.0℃. 1 H NMR (400MHz, DMSO-d6) δ7.80(d,J=3.9Hz,1H),7.24(s,2H),7.04(d,J=3.9Hz,1H),4.74(s,1H),3.64(s,3H),2.31(s,3H)ppm. 13CNMR(101MHz,DMSO-d6)δ165.9,159.8,158.9,149.7,139.9,125.9,119.6,114.8,107.2,106.9,56.5,52.4,34.7,18.9ppm.HRMS(EI-TOF)Calculated[M] + for C 14 H 11 N3O3S:301.0521,Found301.0519.

[0188] Example 22. Preparation of methyl 2-amino-3-cyano-4-(3-furyl)-6-methyl-4H-pyran-5-carboxylate (A22)

[0189]

[0190] Except that furan-3-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 50%.

[0191] Melting point: 171.1-172.8℃. 1 H NMR (400MHz, DMSO-d6) δ7.59-7.47(m,2H),6.97(s,2H),6.31(s,1H),4.32(s,1H),3.66(s,3H),2.29(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.5,159.7,157.3,144.1,139.5,129.3,120.3,110.0,107.4,56.3,52.1,29.9,18.7ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 12 N2O4:260.0797,Found260.0795.

[0192] Example 23. Preparation of methyl 2-amino-3-cyano-4-(4-methyl-5-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A23)

[0193]

[0194] Except that 4-methyl-thiazole-5-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. Yellow solid, total yield 73%.

[0195] Melting point: 198.6-200.5℃. 1 H NMR (400MHz, DMSO-d6) δ8.85(s,1H),7.08(s,2H),4.77(s,1H),3.60(s,3H),2.38(s,3H),2.30(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.1,158.9,157.4,151.9,148.5,138.0,119.8,107.6,57.4,52.3,31.6,18.7,15.1ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 13 N3O3S:291.0678,Found 291.0680.

[0196] Example 24. Preparation of methyl 2-amino-3-cyano-4-(2-methyl-4-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A24)

[0197]

[0198] Except that 5-methyl-thiazole-3-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 60%.

[0199] Melting point: 194.2-195.4℃. 1 H NMR (400MHz, DMSO-d6) δ7.12(s,1H),6.93(s,2H),4.44(s,1H),3.60(s,3H),2.61(s,3H),2.30(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.5,166.1,159.9,158.2,158.0,120.3,114.3,106.7,56.1,52.1,35.6,19.5,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 13 N3O3S:291.0678,Found 291.0681.

[0200] Example 25. Preparation of methyl 2-amino-3-cyano-4-(2-methyl-5-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A25)

[0201]

[0202] Except that 5-methyl-thiazole-2-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 63%.

[0203] Melting point: 185.9-187.8℃. 1 H NMR (400MHz, DMSO-d6) δ7.36(s,1H),7.14(s,2H),4.66(s,1H),3.66(s,3H),2.60(s,3H),2.30(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.1,165.3,159.6,157.9,144.0,138.9,119.8,107.5,56.9,52.3,32.2,19.5,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 13 N3O3S:291.0678,Found 291.0676.

[0204] Example 26. Preparation of methyl 2-amino-3-cyano-4-(2-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A26)

[0205]

[0206] Except that thiazole-2-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 69%.

[0207] Melting point: 201.3-202.7℃. 1 H NMR (400MHz, DMSO-d6) δ7.71(d,J=3.3Hz,1H),7.62(d,J=3.3Hz,1H),7.18(s,2H),4.79(s,1H),3.62(s,3H),2.35(s,3H)ppm. 13 CNMR(101MHz,DMSO-d6)δ173.9,166.1,160.0,159.1,143.1,120.6,119.8,106.4,55.8,52.3,37.3,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 12 H 11N3O3S:277.0521,Found 277.0519.

[0208] Example 27. Preparation of methyl 2-amino-3-cyano-4-(5-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A27)

[0209]

[0210] Except that thiazole-5-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. Yellow solid, total yield 71%.

[0211] Melting point: 166.4-168.3℃. 1 H NMR (400MHz, DMSO-d6) δ8.97(s,1H),7.67(s,1H),7.16(s,2H),4.77(s,1H),3.65(s,3H),2.31(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.0,159.6,158.1,154.1,144.7,140.3,119.8,107.5,56.9,52.3,32.0,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 12 H 11 N3O3S:277.0521,Found 277.0524.

[0212] Example 28. Preparation of methyl 2-amino-3-cyano-4-(4-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A28)

[0213]

[0214] Except that thiazole-3-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 58%.

[0215] Melting point: 206.2-207.0℃. 1 H NMR (400MHz, DMSO-d6) δ8.99(s,1H),7.38(s,1H),6.95(s,2H),4.56(s,1H),3.58(s,3H),2.31(s,3H)ppm. 13C NMR(101MHz,DMSO-d6)δ166.5,160.0,159.5,158.2,154.9,120.3,114.8,106.7,56.1,52.1,35.4,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 12 H 11 N3O3S:277.0521,Found 277.0518.

[0216] Example 29. Preparation of methyl 2-amino-3-cyano-4-(2-bromo-4-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A29)

[0217]

[0218] Except that 5-bromo-thiazole-3-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 62%.

[0219] Melting point: 168.7-170.6℃. 1 H NMR (400MHz, DMSO-d6) δ7.43(s,1H),7.04(s,2H),4.49(s,1H),3.61(s,3H),2.32(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.3,160.0,158.9,158.6,135.8,120.1,119.9,106.0,55.5,52.2,35.7,18.9ppm.HRMS(EI-TOF)Calculated[M] + for C 12 H 10 79 BrN3O3S:354.9626,Found 354.9623.

[0220] Example 30. Preparation of methyl 2-amino-3-cyano-4-(4-bromo-2-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A30)

[0221]

[0222] Except that 4-bromo-thiazole-2-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 72%.

[0223] Melting point: 196.2-198.1℃.1 H NMR (400MHz, DMSO-d6) δ7.75(s,1H),7.28(s,2H),4.78(s,1H),3.64(s,3H),2.37(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ175.5,165.9,160.1,159.7,123.9,119.5,119.0,105.6,55.2,52.4,37.5,18.9ppm.HRMS(EI-TOF)Calculated[M] + for C 12 H 10 79 BrN3O3S:354.9626,Found 354.9623.

[0224] Example 31. Preparation of methyl 2-amino-3-cyano-4-(2-bromo-5-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A31)

[0225]

[0226] Except that 5-bromo-thiazole-2-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 65%.

[0227] Melting point: 179.6-181.4℃. 1 H NMR (400MHz, DMSO-d6) δ7.46(s,1H),7.25(s,2H),4.73(s,1H),3.68(s,3H),2.31(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ165.9,159.8,158.8,148.8,140.0,135.0,119.6,106.7,56.0,52.4,32.4,18.9ppm.HRMS(EI-TOF)Calculated[M] + for C 12 H 10 79 BrN3O3S:354.9626,Found 354.9622.

[0228] Example 32. Preparation of methyl 2-amino-3-cyano-4-(4,5-dimethyl-2-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A32)

[0229]

[0230] Except that 4,5-dimethyl-thiazole-2-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 61%.

[0231] Melting point: 188.0-189.7℃. 1 H NMR (400MHz, DMSO-d6) δ7.14(s,2H),4.60(s,1H),3.63(s,3H),2.34(s,3H),2.28(s,3H),2.20(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ168.8,166.1,159.9,158.8,148.1,126.6,119.8,106.4,55.8,52.3,37.2,18.8,15.1,11.4ppm.HRMS(EI-TOF)Calculated[M] + for C 14 H 15 N3O3S:305.0834,Found305.0829.

[0232] Example 33. Preparation of methyl 2-amino-3-cyano-4-(2-benzothiazolyl)-6-methyl-4H-pyran-5-carboxylate (A33)

[0233]

[0234] Except that benzothiazole-2-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 63%.

[0235] Melting point: 207.4-209.2℃. 1 H NMR (400MHz, DMSO-d6) δ8.05(d,J=8.0Hz,1H),7.97(d,J=8.1Hz,1H),7.47(m,2H),7.29(s,2H),4.86(s,1H),3.60(s,3H),2.39(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ175.1,166.0,160.0,159.7,153.3,135.2,126.8,125.7,12 3.3,122.9,119.6,105.7,55.0,52.5,38.3,19.0ppm.HRMS(EI-TOF)Calculated[M] + for C16 H 13 N3O3S:327.0678,Found 327.0675.

[0236] Example 34. Preparation of methyl 2-amino-3-cyano-4-(3,4-dichloro-5-isothiazolyl)-6-methyl-4H-pyran-5-carboxylate (A34)

[0237]

[0238] Add 3,4-dichloro-isothiazole-5-carboxylic acid (2.00 g, 10.00 mmol, 1.0 eq.) and 20 mL of anhydrous tetrahydrofuran to a 100 mL eggplant-shaped bottle under ice bath conditions, then add oxalyl chloride (3.20 g, 25.00 mmol, 2.5 eq.) and 2 drops of DMF. After reacting for 1 h, concentrate under reduced pressure to obtain 3,4-dichloro-isothiazole-5-carboxylic acid chloride. Add 10 mL of methanol and continue the reaction for 30 min. After the reaction is completed, concentrate under reduced pressure to obtain 3,4-dichloro-isothiazole-5-carboxylic acid methyl ester, which can be directly reacted in the next step without purification.

[0239]

[0240] Dissolve 3,4-dichloro-isothiazole-5-carboxylic acid methyl ester (2.20 g, 10.00 mmol, 1.0 eq.) in 20 mL methanol in a 100 mL eggplant-shaped bottle, add sodium borohydride (1.50 g, 40.00 mmol, 4.0 eq.) in batches under ice bath conditions, and continue the reaction for 2 h. After TLC shows that the reaction is complete, add dilute hydrochloric acid (1 mol·L -1 ) until pH = 2, then add saturated NaHCO3 solution to pH = 8, remove methanol under reduced pressure, add ethyl acetate to extract and separate the liquids three times, combine the organic phases, dry over anhydrous Na2SO4 and concentrate. The crude product was separated by column chromatography (petroleum ether: ethyl acetate = 3:1, v / v) to obtain 3,4-dichloro-isothiazole-5-methanol, 1.60 g of white solid, with a yield of 87%.

[0241] 1 H NMR (400MHz, CDCl3) δ4.98 (d, J = 5.44Hz, 2H), 2.51 (t, J = 5.82Hz, 1H) ppm.

[0242] Dissolve 3,4-dichloro-isothiazole-5-methanol (1.60 g, 8.70 mmol, 1.0 eq.) in 25 mL of dichloromethane in a 100 mL eggplant-shaped bottle, add Dess-Martin reagent (5.50 g, 13.00 mmol, 1.5 eq.) in batches under ice bath conditions, and continue the reaction for 2 h. After TLC shows that the reaction is complete, add saturated NaHCO3 solution dropwise until no bubbles are generated, and then add saturated Na2S2O3 solution to quench. Filter to remove solid salts, separate the liquids to obtain dichloromethane phase, add ethyl acetate to the aqueous phase for extraction and separation 3 times, combine the organic phases, dry with anhydrous Na2SO4 and concentrate. The crude product is separated by column chromatography (petroleum ether: ethyl acetate = 5:1, v / v) to obtain 3,4-dichloro-isothiazole-5-carboxaldehyde, 1.10 g of white solid, with a yield of 70%.

[0243] 1 H NMR (400MHz, CDCl3) δ10.07 (s, 1H) ppm.

[0244]

[0245] Except that 3,4-dichloro-isothiazole-5-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 75%.

[0246] Melting point: 185.1-186.9℃. 1 H NMR (400MHz, DMSO-d6) δ7.35(s,2H),4.96(s,1H),3.62(s,3H),2.37(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ168.5,165.5,160.2,159.9,147.8,119.2,118.9,103.9,54.0,52.5,33.0,18.9ppm.HRMS(EI-TOF)Calculated[M] + forC 12 H9 35 Cl2N3O3S:344.9742,Found 344.9739.

[0247] Example 35. Preparation of methyl 2-amino-3-cyano-4-(4-methyl-5-1,2,3-thiadiazolyl)-6-methyl-4H-pyran-5-carboxylate (A35)

[0248]

[0249] In an ice bath, add 4-methyl-1,2,3-thiadiazole-5-carboxylic acid (1.50 g, 10.00 mmol, 1.0 eq.) and 20 mL of anhydrous tetrahydrofuran to a 100 mL eggplant-shaped bottle, then add oxalyl chloride (3.20 g, 25.00 mmol, 2.5 eq.) and 2 drops of DMF. After reacting for 1 h, concentrate under reduced pressure to obtain 4-methyl-1,2,3-thiadiazole-5-carboxylic acid chloride. Add 10 mL of methanol and continue the reaction for 30 min, then concentrate under reduced pressure to obtain 4-methyl-1,2,3-thiadiazole-5-carboxylic acid methyl ester, which can be directly used for the next step without purification.

[0250]

[0251] Dissolve 4-methyl-1,2,3-thiadiazole-5-carboxylic acid methyl ester (1.60 g, 10.00 mmol, 1.0 eq.) in 20 mL of methanol in a 100 mL eggplant-shaped bottle, add sodium borohydride (1.50 g, 40.00 mmol, 4.0 eq.) in batches under ice bath, and continue the reaction for 2 h. After TLC shows that the reaction is complete, add dilute hydrochloric acid (1 mol·L -1 ) until pH = 2, then add saturated NaHCO3 solution until no more bubbles are generated, remove methanol under reduced pressure, add ethyl acetate to extract and separate the liquids 5 times, combine the organic phases, dry over anhydrous Na2SO4 and concentrate. The crude product is separated by column chromatography (petroleum ether: ethyl acetate = 1:1, v / v) to obtain 4-methyl-1,2,3-thiadiazole-5-methanol, 1.15 g of yellow oily liquid, with a yield of 89%.

[0252] 1 H NMR (400MHz, CDCl3) δ5.01 (s, 2H), 2.62 (s, 3H) ppm.

[0253] Dissolve 4-methyl-1,2,3-thiadiazole-5-methanol (1.15g, 8.90mmol, 1.0eq.) in 25mL dichloromethane in a 100mL eggplant-shaped bottle, add Dess-Martin reagent (5.60g, 13.30mmol, 1.5eq.) in batches under ice bath, and continue to react for 2h. After TLC shows that the reaction is complete, add saturated NaHCO3 solution dropwise until no bubbles are generated, and then add saturated Na2S2O3 solution to quench. Filter to remove solid salts, separate the liquid to obtain dichloromethane phase, add ethyl acetate to the aqueous phase, extract and separate the liquid 3 times, combine the organic phases, dry with anhydrous Na2SO4 and concentrate. The crude product is separated by column chromatography (petroleum ether: ethyl acetate = 3:1, v / v) to obtain 4-methyl-1,2,3-thiadiazole-5-carboxaldehyde, 821mg of light yellow oily liquid, and the yield is 72%.

[0254] 1H NMR (400MHz, CDCl3) δ10.26 (s, 1H), 3.04 (s, 3H) ppm.

[0255]

[0256] Except that 4-methyl-1,2,3-thiadiazole-5-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 70%.

[0257] Melting point: 171.3-173.0℃. 1 H NMR (400MHz, DMSO-d6) δ7.28(s,2H),5.02(s,1H),3.58(s,3H),2.66(s,3H),2.35(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ165.7,159.1,158.8,157.9,156.0,119.4,106.4,56.0,52.4,30.4,18.9,12.4ppm.HRMS(EI-TOF)Calculated[M] + for C 12 H 12 N4O3S:292.0630,Found 292.0627.

[0258] Example 36. Preparation of methyl 2-amino-3-cyano-4-((2-methyl-4-trifluoromethyl-5-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A36)

[0259]

[0260] Except that 2-methyl-4-trifluoromethyl-thiazole-5-carboxylic acid was used to replace 3,4-dichloro-isothiazole-5-carboxylic acid in Example 34, other conditions and steps were the same as those in Example 34. Colorless oily liquid, the total yield of the four-step reaction was 62%.

[0261] 1 H NMR (400MHz, CDCl3) δ10.34(s,1H),3.37(s,3H)ppm.

[0262]

[0263] Except that 2-methyl-4-trifluoromethyl-thiazole-5-carboxaldehyde was used to replace p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 78%.

[0264] Melting point: 209.7-211.1℃. 1 H NMR (400MHz, DMSO-d6) δ7.27(s,1H),4.96(s,1H),3.58(s,3H),2.64(s,3H),2.35(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.8,165.6,159.1,158.7,149.0,136.3(q, 2 J C-F =34.3Hz),121.7(q, 1 J C-F =272.7Hz),118.9,107.2,56.7,52.3,31.7,19.4,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 14 H 12 F3N3O3S:359.0551,Found 359.0550.

[0265] Example 37. Preparation of methyl 2-amino-3-cyano-4-(2,4-dichloro-5-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A37):

[0266]

[0267] Except that 2,4-dichloro-thiazole-5-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 31%.

[0268] Melting point: 197.4-199.2℃. 1 H NMR (400MHz, DMSO-d6) δ7.26(s,2H),4.77(s,1H),3.61(s,3H),2.31(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ165.6,159.4,159.2,149.4,139.8,132.8,119.3,105.1,54.9,52.5,32.1,18.9ppm.HRMS(EI-TOF)Calculated[M] + forC 12 H9 35 Cl2N3O3S:344.9742,Found 344.9745.

[0269] Example 38. Preparation of methyl 2-amino-3-cyano-4-(4-trifluoromethyl-5-thiazolyl)-6-methyl-4H-pyran-5-carboxylate (A38)

[0270]

[0271] Add 2-amino-4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester (2.00g, 8.32mmol, 1.0eq.) and 20mL 1,4-dioxane to a 100mL two-necked flask at room temperature, connect a condenser and protect with a nitrogen atmosphere. Slowly add tert-butyl nitrite (1.30g, 12.50mmol, 1.5eq.) dropwise under ice bath conditions, and then heat to 90℃ to continue the reaction for 5h. After TLC shows that the reaction is complete, remove the solvent under reduced pressure, and the crude product is separated by column chromatography (petroleum ether: ethyl acetate = 5:1, v / v) to obtain 4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester, 1.35g of yellow-green solid, with a yield of 72%.

[0272] 1 H NMR (400MHz, CDCl3) δ8.87 (s, 1H), 4.40 (q, J = 7.1Hz, 2H), 1.38 (t, J = 7.1Hz, 3H) ppm.

[0273] Add 4-trifluoromethyl-thiazole-5-carboxylic acid ethyl ester (1.35 g, 6.00 mmol, 1.0 eq.) and 20 mL of anhydrous dichloromethane to a 100 mL two-necked bottle at room temperature, and cool to -78°C under nitrogen atmosphere. Slowly add 1 mol·L -1 Diisobutylaluminum hydride n-hexane solution (6.60mL, 6.60mmol, 1.1eq.) was added dropwise and the reaction was continued for 1h. After TLC showed that the reaction was complete, 0.35mL of water and 0.35mL of 15% NaOH aqueous solution were added in sequence to quench the reaction solution, and then a small amount of anhydrous Na2SO4 was added and stirred at room temperature for 10min, and then diatomaceous earth was padded and filtered to obtain the filtrate. The crude product was separated by column chromatography (petroleum ether: ethyl acetate = 5:1, v / v) to obtain 4-trifluoromethyl-thiazole-5-carboxaldehyde, 326mg of yellow oily liquid, with a yield of 30%.

[0274] 1 H NMR (400MHz, CDCl3) δ10.45 (s, 1H), 8.89 (s, 1H) ppm.

[0275]

[0276] Except that 4-trifluoromethyl-thiazole-5-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 33%.

[0277] Melting point: 198.8-199.9℃. 1 H NMR (400MHz, DMSO-d6) δ9.15(s,1H),7.29(s,2H),5.05(s,1H),3.57(s,3H),2.36(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ165.6,159.0,158.8,155.8,149.9,138.0(q, 2 J C-F =34.3Hz),121.9(q, 1 J C-F =272.7Hz),118.9,107.2,56.8,52.3,31.6,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 13 H 10 F3N3O3S:345.0395,Found345.0392.

[0278] Example 39. Preparation of methyl 2-amino-3-cyano-4-(1-methyl-1,2,3-triazolyl)-6-methyl-4H-pyran-5-carboxylate (A39)

[0279]

[0280] Except that 1-methyl-1,2,3-triazole-4-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 70%.

[0281] Melting point: 196.0-198.0℃. 1 H NMR (400MHz, DMSO-d6) δ7.85(s,1H),6.96(s,2H),4.50(s,1H),4.00(s,3H),3.61(s,3H),2.31(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.4,159.8,158.0,150.4,123.0,120.3,106.6,56.1,52.2,36.7,30.8,18.8ppm.HRMS(EI-TOF)Calculated[M] + for C 12 H 13 N5O3:275.1018,Found 275.1015.

[0282] Example 40. Preparation of methyl 2-amino-3-cyano-4-(3,5-dimethyl-4-isoxazolyl)-6-methyl-4H-pyran-5-carboxylate (A40)

[0283]

[0284] Except that 3,5-dimethyl-isoxazole-4-carboxaldehyde was used to replace the p-trifluoromethylbenzaldehyde in Example 1, other conditions and steps were the same as those in Example 1. White solid, total yield 80%.

[0285] Melting point: 194.0-195.3℃. 1 H NMR (400MHz, DMSO-d6) δ6.84(s,2H),4.20(s,1H),3.43(s,3H),2.13(s,3H),2.12(s,3H),1.90(s,3H)ppm. 13 C NMR(101MHz,DMSO-d6)δ166.3,166.0,159.1,158.8,157.6,119.8,116.3,104.8,54.7,52.2,28.1,18.6,10.8,9.9ppm.HRMS(EI-TOF)Calculated[M] + for C 14 H 15 N3O4:289.1063,Found 289.1059.

[0286] Activity Test Example

[0287] 1. Anti-disease activity test part of the present invention

[0288] 1.1 Active oxygen burst level test

[0289] Reactive oxygen species (ROS) are the products of oxygen gaining electrons, including four types: singlet oxygen, superoxide anion, H2O2 and hydroxyl free radicals. The burst of reactive oxygen species is one of the important immune responses of plants to resist disease infection. This allergic reaction often occurs in the early stage of the plant immune process. When plants are stimulated by external harmful conditions, ROS are overexpressed and accumulated in the plant body, triggering an immune response to directly kill pathogenic microorganisms. ROS can also be used as an important signal molecule to regulate the redox level in plant cells and induce plants to trigger stomatal closure and immune response. Plant immune response is always accompanied by the generation and release of reactive oxygen species. Under normal growth conditions, ROS, as a byproduct of oxygen metabolism, is stably maintained at a low level. If plant immune activators are used before the disease occurs, the ROS level in the plant body can be significantly increased, inducing enhanced disease resistance and thus achieving the effect of preventing and controlling diseases. Therefore, quantitative detection of ROS levels can intuitively reflect the activity of the compound to be tested. Among them, diaminobenzidine (DAB) is a common peroxidase colorimetric agent. The generated superoxide anions and H2O2 oxidize DAB to form a brown-red precipitate at the active site of the enzyme. The reaction product is insoluble in water and organic solvents and has good stability and anti-interference properties.

[0290] At the same time, the inventors developed a recognition algorithm software to calculate the total leaf area and sediment area according to the color blocks, and further obtain the percentage of sediment, so as to judge the ability and activity of the compound to induce ROS. Multiple experiments have proved that this method is quite reliable and can be used as a preliminary screening platform for compound activity.

[0291] Experimental object: Active oxygen burst level

[0292] Test concentration: All compounds were tested at 100 mg·L -1 Carried out at concentration.

[0293] Positive control: The positive control agent selected was the commercial plant immune activator flutolide (FBT) successfully developed by the inventor's team, which acts on the dual signaling pathways of salicylic acid and jasmonic acid, and has obtained a temporary license for new pesticide registration in 2015, and can effectively prevent and control a variety of plant diseases.

[0294] Test method: Sow various crops in advance, weigh the compound samples quantitatively, dissolve them with an appropriate amount of DMSO, add an appropriate amount of surfactant, and dilute them with deionized water to the set concentration for use. Prepare a certain concentration of diaminobenzidine colorimetric working solution in advance and store it in a dark place for use. Spray the compound solution to be tested evenly on both sides of the crop leaves, and then induce them under constant conditions for a period of time. After the compound induction is completed, soak each group of leaves with an appropriate amount of diaminobenzidine solution for a period of time. Rinse the leaves with deionized water, boil the leaves with a certain concentration of ethanol solution to remove chlorophyll, and finally form an insoluble reddish-brown precipitate on the surface of the leaves.

[0295] Calculation of results: Each group of leaves was placed on a background plate and photographed. The percentage of the reddish-brown precipitate on the leaf surface was calculated using software. After deducting the water group control, the relative ability of the compound to induce the production of reactive oxygen species was calculated.

[0296] The test was carried out according to the above method. From the total of 40 compounds in this series, eight compounds with better initial screening activity in each group were selected for further verification of the active oxygen burst test.

[0297] According to the above method, the active oxygen burst test data of the preferred compound is shown in the following Table 1:

[0298] Table 1: Active oxygen burst test results of eight selected compounds

[0299]

[0300]

[0301] Analysis and discussion: The retest results of active oxygen burst showed that the series of compounds basically maintained their original activity, among which compounds A1, A23, A34, and A36 showed higher activity. The creation of plant immune activators requires the construction of a stable and reliable activity evaluation system. Active oxygen is a key signaling molecule in the plant immune system, so the quantitative detection of active oxygen levels is very important. Based on the active oxygen burst test platform established by our team, as a preliminary activity evaluation method, after quickly and easily obtaining compounds with certain activity, plant-level plant immune activation activity tests can be carried out, which can greatly improve the efficiency of new structure discovery and screening.

[0302] 1.2 In vivo efficacy test

[0303] Experimental object: Cucumber gray mold

[0304] Test concentration: All compounds were tested at four concentrations of 0.1 mg·L -1 , 1mg·L -1 、10mg·L -1、100mg·L -1 Next proceed.

[0305] Positive control: The test used the clean water group as the negative control, and selected fluoborbide activated ester (FBT) and jinggangmycin as positive control agents. Jinggangmycin was discovered in the 1970s and is a highly effective and low-toxic antibiotic produced by the metabolism of actinomycetes. Plant immune activators have different mechanisms of action from traditional fungicides and need to be used at appropriate concentrations to achieve better control effects. Therefore, four concentrations were used for testing at the same time. According to previous studies, the most suitable concentration of FBT is 0.1 mg·L -1 The most suitable concentration of Jinggangmycin is 100 mg·L -1 .

[0306] Test method: Sow various crops in advance, weigh the compound samples quantitatively, dissolve them with an appropriate amount of DMSO, add an appropriate amount of surfactant, and dilute with water to the set concentration. When the crops grow to a certain period, use the backward method to treat the plants with drugs three times, 7 days, 4 days, and 1 day before inoculation of pathogens, and then inoculate pathogens at the same time. The experiment was carried out using the potted method and repeated 3 times. The calculation method of disease index and disease prevention effect is as follows:

[0307] Disease index = [∑(number of diseased leaves at each level × relative level value) × 100] / (total number of leaves surveyed × representative value of the highest level of disease).

[0308] Control effect (%) = [(disease index of control area - disease index of treated area) × 100] / disease index of control area.

[0309] According to the above method, the in vivo efficacy test data of the preferred compounds are shown in the following Table 2:

[0310] Table 2: Control effect of eight selected compounds on cucumber gray mold

[0311]

[0312]

[0313]

[0314] Analysis and discussion: The test found that compound A34 showed similar activity at three concentrations, among which at 100 mg·L -1 The activity was higher than that of A36, which was consistent with the test results of active oxygen level. -1 When the concentration was further reduced to 0.1 mg·L -1The activity of compound A34 was slightly higher than that of A36. -1 The highest control efficiency against cucumber gray mold was 58.43% at the concentration of 1.5 mg·L, but there was still a certain gap between the activity and the control agent. The most suitable concentration for most compounds was 0.1 mg·L -1 and 1mg·L -1 , generally better than high concentration 10mg·L -1 and 100mg·L -1 The reason is speculated to be related to the mechanism of action of plant immune activators, which cause an increase in the activity of plant defense-related enzymes, including polyphenol oxidase, phenylalanine ammonia lyase, superoxide dismutase, etc., resulting in changes in physiological and biochemical mechanisms, thereby enhancing the disease resistance of the plant. If the dosage concentration is too high, it will inhibit the activity of such enzymes, resulting in poor prevention effect at high concentrations. Plant immune activators can also cause the accumulation of pathogenesis-related proteins, exert anti-disease effects by attacking pathogens and degrading toxins, and induce plants to produce overall disease resistance. Similarly, the expression of such proteins is inhibited at high concentrations.

[0315] 1.3 In vitro antibacterial activity test

[0316] One of the notable characteristics of plant immune activators is that the compounds show high antibacterial effects under in vivo conditions, but have almost no direct antibacterial effect under in vitro conditions. Therefore, after confirming that the series of compounds have in vivo antibacterial effects, the compounds with higher in vivo antibacterial effects are tested for in vitro antibacterial activity to confirm that they play the role of true plant immune activators. As shown in Table 2, most of the compounds in this series have a high antibacterial effect at 1 mg·L -1 The concentration of 100 mg·L -1 No significant activity was shown at all concentrations. -1 This is a more appropriate concentration for use of this series of compounds, and subsequent in vitro antibacterial activity tests were also carried out at this concentration.

[0317] Test objects: Cucumber gray mold, rice sheath blight, rapeseed sclerotinia, wheat fusarium

[0318] Test concentration: All compounds were tested at 1 mg·L -1 concentration.

[0319] Positive control: The commercial nicotinamide broad-spectrum fungicide boscalid was selected as the positive control agent.

[0320] Test method: The mycelium growth rate method was used for determination. After the compound was dissolved in an appropriate amount of DMSO, a small amount of surfactant was added and diluted with water to prepare 50 mg·L -1Then take 1 mL of the compound stock solution and mix it thoroughly with 49 mL of PDA medium solution to make the concentration of the compound to be tested 1 mg·L -1 , and then pour it evenly into 3 90mm culture dishes. After solidification, inoculate pathogens in the culture dishes respectively; after cultivation, measure the average growth diameter of the colonies by the cross method, compare it with the blank control group, and calculate the inhibition rate of the compound on pathogens under in vitro conditions.

[0321] The test was carried out according to the above method, and the test results of the in vitro antibacterial activity of the preferred compounds are shown in Table 3 below.

[0322] Table 3: In vitro antibacterial activity of eight preferred compounds

[0323]

[0324]

[0325] Among them, BC: cucumber gray mold; SS: rapeseed sclerotinia; RS: rice sheath blight; FG: wheat fusarium.

[0326] Analysis and discussion: The in vitro antibacterial activity test showed that the antibacterial activity of this series of compounds was lower than that of the control drug boscalid. At the same time, the inhibitory effect on cucumber gray mold was also significantly lower than the corresponding plant immune activation activity at the same concentration, proving that it played an immune activation role rather than a direct bactericidal effect.

[0327] The results of in vivo immune activation and in vitro antibacterial activity tests show that this series of compounds has a high protective effect against pathogens under in vivo conditions, but almost no direct antibacterial activity under in vitro conditions. Therefore, 2-amino-3-cyano-4-aryl-4H-pyran compounds are expected to be developed into new plant immune activators.

[0328] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound of formula I, or an optical isomer, cis-trans isomer or pharmaceutically acceptable salt thereof, wherein R1 is selected from the group consisting of substituted or unsubstituted C5-C8 aryl (preferably phenyl), substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C3-C6 epoxyalkyl, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 alkylthio, substituted or unsubstituted C1-C6 alkylsulfonyl, substituted or unsubstituted C l -C6 alkyl acyl, substituted or unsubstituted C l -C6 alkyl ester group, substituted or unsubstituted C l -C6 alkylamide; R2 is selected from the following group: substituted or unsubstituted C5-C8 aryl (preferably phenyl), substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 epoxyalkyl, substituted or unsubstituted C1-C6 alkylamino, substituted or unsubstituted C1-C6 alkylthio, substituted or unsubstituted C1-C6 alkylsulfonyl, substituted or unsubstituted C l -C6 alkyl acyl, substituted or unsubstituted C l -C6 alkyl ester group, substituted or unsubstituted C l -C6 alkylamide; Ar is selected from the following group: a substituted or unsubstituted monocyclic, bicyclic or tricyclic aromatic group containing 6 to 14 carbon atoms, or a substituted or unsubstituted 5 to 10 membered monocyclic, bicyclic or tricyclic heteroaromatic group containing 1 to 3 heteroatoms selected from N, O or S.

2. The compound according to claim 1, characterized in that In the formula, R1 is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C6 cycloalkyl; R2 is selected from the group consisting of substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy; Ar is selected from the group consisting of substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, or substituted or unsubstituted 5-10 membered monocyclic or bicyclic heteroaryl.

3. The compound according to claim 2, characterized in that R1 is a substituted or unsubstituted C1-C4 alkyl group; R2 is a substituted or unsubstituted C1-C4 alkyl group; Ar is a substituted or unsubstituted phenyl group, or a substituted or unsubstituted 5-10 membered heteroaryl group.

4. The compound according to claim 3, characterized in that Ar is phenyl, furanyl, thienyl, thiazolyl, isoxazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl or isothiazolyl, and is optionally substituted by 1-3 substituents selected from the group consisting of halogen, nitro, cyano, methyl, methoxy, methylthio, trifluoromethyl; Preferably, Ar is phenyl, thiazolyl or isothiazolyl, and is optionally substituted by 1-2 substituents selected from the group consisting of halogen, methyl, trifluoromethyl; Preferably, Ar is thiazolyl or isothiazolyl, and is optionally substituted by 1-2 substituents selected from the group consisting of halogen, methyl, trifluoromethyl.

5. A compound selected from the group consisting of:

6. The compound according to claim 5, characterized in that The compound is selected from the group consisting of: Preferably, the compound is selected from the group consisting of:

7. A pesticide composition, characterized in that: The pesticide composition contains: (1) 0.001-99.99 wt% of the total weight of the pharmaceutical composition of the compound of any one of claims 1-6, or its optical isomers, cis-trans isomers or pesticide-acceptable salts; and (2) A pesticide-acceptable carrier or excipient.

8. Use of the compound according to any one of claims 1 to 6, or its optical isomers, cis-trans isomers or pesticide-acceptable salts in the preparation of pesticides.

9. A method for preventing and controlling plant diseases, characterized in that: The method comprises: administering an effective amount of the compound according to any one of claims 1 to 6 or the pesticide composition according to claim 7 to the plant, thereby preventing and controlling the corresponding disease.

10. The method for preparing the compound according to any one of claims 1 to 6, characterized in that: The compound was prepared using the general synthetic route shown below: Wherein, R1, R2 and Ar are as described in any one of claims 1-6.

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