A benzisothiazole carboxamide compound and application thereof

Abstract

The application discloses a benzisothiazole carboxamide compound and application thereof. The benzisothiazole carboxamide compound has the characteristics of high specificity and high activity as an inhibitor of myosin motor domain, and can significantly inhibit the growth and development process and pathogenicity of pathogenic fungi, especially Pyricularia oryzae.

Description

Technical Field

[0001] This invention relates to the field of agricultural disease control technology, and more specifically, to a benzisothiazolium carboxamide compound and its applications. Background Technology

[0002] Fungi are a large group of eukaryotes, and their strong survival ability allows them to be widely distributed in nature. Fungal diseases are the largest category of plant diseases, affecting a wide area and causing extremely serious damage. Currently, control mainly relies on chemical control methods, but existing chemical control methods have problems such as single target, serious structural homogeneity, and constantly increasing resistance, making the development of new targets for green pesticides urgent. Finding new targets can lead to the development of green pesticides with entirely new mechanisms of action, breaking through the limitations of existing agents. These new pesticides can not only effectively address the resistance of pathogenic fungi, but also reduce negative environmental impacts, achieving sustainable agricultural development.

[0003] Rice ( Oryza sativa Rice is one of the world's most important food crops. Rice blast is caused by the fungus *Magnapordia oryzae* (…). Magnaporthe oryzae This disease, caused by [unspecified pathogen], is considered by plant pathologists to be the most devastating of the ten most harmful crop fungal diseases globally. It is widely distributed in all major rice-growing regions worldwide, severely impacting rice yield and quality.

[0004] Fungal cell walls are rich in chitin, glucose, and mannose proteins, with chitin being an important and common cell wall component of fungi. Chitin synthases (Chs) are key enzymes in chitin synthesis. Fungal chitin synthases can be classified into seven classes, each containing a chitin synthase domain (CSD) and multiple transmembrane domains (TM). Furthermore, classes 5 and 6 chitin synthases both contain a myosin motor domain (MMD) at their N-terminus. This MMD belongs to class 17 of myosin domains, is found only in filamentous fungi, and contributes to the virulence of various pathogens.

[0005] The rice blast fungus's Chs5 and Chs6 belong to the fifth and sixth classes of chitin synthases, respectively, and both contain an MMD at the N-terminus. Phenotypically, chs5 Knockout does not affect pathogenicity. chs6 Knockout cells affect pathogenicity. chs5 / chs6 Double knockout completely eliminates pathogenicity. chs6△MMD The knockout form also completely loses its pathogenicity. CHS5 and CHS6The gene expression level was highest in the vegetative hyphae. The MMD of Chs6 contains highly conserved P-loop, Switch I, and Switch II myosin domains, while the MMD of Chs5 lacks these structures. This difference in the MMD between Chs5 and Chs6 may explain their functional differences. These results indicate that the MMDs of Chs5 and Chs6 play a crucial role in the infection of rice by *Blastomyces oryzae* and may represent novel fungicide targets. Summary of the Invention

[0006] This invention is based on the inventor's discovery and understanding of the following facts and problems: the control of fungal diseases mainly relies on chemical control methods, but current chemical control methods have problems such as single target, serious structural homogenization, and continuous enhancement of resistance.

[0007] This invention aims to at least partially solve one of the technical problems in related technologies. To this end, embodiments of this invention propose a benzisothiazolium carboxamide compound and its application. As an inhibitor of the myosin motor domain, the benzisothiazolium carboxamide compound exhibits high specificity and activity, and can significantly inhibit the growth and development of pathogenic fungi, especially rice blast fungus, and its pathogenicity.

[0008] This invention provides a benzisothiazolium formamide compound, the structural formula of which is shown below:

[0009] .

[0010] The advantages and technical effects of the benzisothiazolium carboxamide compounds in the embodiments of the present invention are as follows: benzisothiazolium carboxamide compounds can act as inhibitors of the myosin motor domain (MMD), and have the characteristics of high specificity and high activity, and can significantly inhibit the growth and development process and pathogenicity of pathogenic fungi (especially rice blast fungus).

[0011] This invention provides a method for preparing benzisothiazolium carboxamide compounds, comprising the following steps:

[0012] (1) Compound 1 was reacted with methylamine hydrochloride to obtain compound 2;

[0013] (2) The compound 2 was mixed with a 1,4-dioxane solution of hydrochloric acid and reacted to obtain compound 3;

[0014] (3) The compound 3 was reacted with 5-bromobenzo[d]isothiazol-3-carboxylic acid to give compound 4;

[0015] (4) The compound 4 was reacted with 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole to obtain benzisothiazol carboxamide compounds;

[0016] The structural formulas of compounds 1, 2, 3, and 4 are shown below:

[0017] .

[0018] In some embodiments, in step (1), the reaction is carried out under the conditions of benzotriazole-1-yloxy-tris(tetrahydropyrrolidinyl)phosphonium hexafluorophosphate and N,N-diisopropylethylamine;

[0019] And / or, the molar ratio of compound 1 to methylamine hydrochloride is 1:1;

[0020] And / or, the temperature of the reaction is 25°C;

[0021] And / or, the reaction time is 1 h;

[0022] And / or, in step (2), the reaction temperature is 25°C;

[0023] And / or, the reaction time is 1 h.

[0024] In some embodiments, in step (3), the molar ratio of compound 3 and 5-bromobenzo[d]isothiazol-3-carboxylic acid is 1.16:0.39;

[0025] And / or, the reaction is carried out under the conditions of benzotriazole-1-yloxy-tris(tetrahydropyrrolidinyl)phosphonium hexafluorophosphate and N,N-diisopropylethylamine;

[0026] And / or, the temperature of the reaction is 25°C;

[0027] And / or, the reaction time is 1 h.

[0028] In some embodiments, in step (4), the molar ratio of compound 4 and 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole is 1:1;

[0029] And / or, the reaction is carried out in aqueous solution of chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) and potassium carbonate;

[0030] And / or, the temperature of the reaction is 100°C;

[0031] And / or, the reaction time is 1 h;

[0032] And / or, the reaction is carried out under a protective atmosphere.

[0033] This invention provides a myosin motor domain inhibitor, comprising the benzisothiazolium carboxamide compound or its pesticide-acceptable salt as described in this invention.

[0034] In this embodiment of the invention, benzisothiazolium carboxamide compounds, as inhibitors of the myosin motor domain, have the characteristics of high specificity and high activity, and can significantly inhibit the growth and development process and pathogenicity of pathogenic fungi (especially rice blast fungus).

[0035] In some embodiments, a pesticide-acceptable carrier or excipient may also be included;

[0036] And / or, the dosage form of the myosin motor domain inhibitor includes at least one of emulsifiable concentrate, wettable powder, suspension concentrate, soluble powder, aqueous solution, fumigant, granule or seed coating agent;

[0037] And / or, the myosin motor domain is derived from chitin synthase of pathogenic microorganisms.

[0038] This invention provides an application of benzisothiazol carboxamide compounds or myosin motor domain inhibitors for the prevention and control of plant diseases caused by pathogenic microorganisms.

[0039] In the embodiments of the present invention, the benzisothiazolium carboxamide compounds and the inhibitors of myosin motor domains are characterized by high specificity and high activity, and can significantly inhibit the growth and development process and pathogenicity of pathogenic fungi (especially rice blast fungus), and can be used to prevent and control plant diseases caused by pathogenic microorganisms.

[0040] In some embodiments, the pathogenic microorganism includes pathogenic fungi;

[0041] And / or, the pathogenic microorganism includes at least one of the myosin motor domain or its homologous protein;

[0042] And / or, the plant diseases include plant fungal diseases.

[0043] In some embodiments, the pathogenic microorganism includes rice blast fungus;

[0044] And / or, the plant disease includes rice blast;

[0045] And / or, the plant includes rice. Attached Figure Description

[0046] Figure 1 This is a graph showing the interaction results between benzisothiazol carboxamide compounds and MMD of rice blast fungus CHS in Example 1, detected by surface plasmon resonance method.

[0047] Figure 2 The effect of benzisothiazolamide compounds on the growth and development of rice blast fungus was tested in Example 1 by inoculating rice blast fungus cakes into the center of CM medium containing or without the compounds.

[0048] Figure 3 This is a graph showing the effect of different concentrations of benzisothiazolamide compounds on the pathogenicity of rice blast fungus and the statistical results of lesion area, as detected by an in vitro inoculation experiment based on spot inoculation of rice leaves. Detailed Implementation

[0049] Embodiments of the present invention are described in detail below, examples of which are illustrated in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain the present invention, and should not be construed as limiting the present invention.

[0050] An embodiment of the present invention provides a benzisothiazolium carboxamide compound, the structural formula of which is shown below:

[0051] .

[0052] The benzisothiazol carboxamide compounds of the present invention can act as inhibitors of the myosin motor domain (MMD), exhibiting high specificity and activity, and can significantly inhibit the growth and development process and pathogenicity of pathogenic fungi (especially rice blast fungus).

[0053] A method for preparing a benzisothiazolium carboxamide compound according to an embodiment of the present invention includes the following steps:

[0054] (1) Compound 1 was reacted with methylamine hydrochloride to obtain compound 2;

[0055] Optionally, in step (1), the reaction is carried out under the conditions of benzotriazole-1-yloxy-tris(tetrahydropyrrolidinyl)phosphonium hexafluorophosphate and N,N-diisopropylethylamine; the molar ratio of compound 1 to benzotriazole-1-yloxy-tris(tetrahydropyrrolidinyl)phosphonium hexafluorophosphate is 1.66:1.99; the molar ratio of compound 1 to N,N-diisopropylethylamine is 1.66:4.98;

[0056] Optionally, the molar ratio of compound 1 to methylamine hydrochloride is 1:1;

[0057] Optionally, the reaction temperature is 25°C; the reaction time is 1 h;

[0058] Optionally, the solvent for the reaction includes dichloromethane;

[0059] In a specific embodiment, step (1) is described by the following formula:

[0060] ;

[0061] In a specific embodiment, benzotriazole-1-yloxy-tris(tetrahydropyrrolyl)phosphonium hexafluorophosphate (PyBOP, 1.04 g, 1.99 mmol) and N,N-diisopropylethylamine (DIPEA or DIEA, 0.643 g, 4.98 mmol, 0.87 mL) were added to a dichloromethane (5 mL) solution of compound 1 (0.500 g, 1.66 mmol) and methylamine hydrochloride (0.112 g, 1.66 mmol), and the mixture was stirred at 25 °C for 1 h.

[0062] Optionally, a post-processing is performed after the reaction, the post-processing comprising: concentrating the reaction mixture under reduced pressure, purifying the residue by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 50%), monitoring by thin-layer chromatography (developing solvent: petroleum ether: ethyl acetate = 1:1, Rf = 0.1), to obtain a white solid compound 2.

[0063] (2) The compound 2 was mixed with a 1,4-dioxane solution of hydrochloric acid and reacted to obtain compound 3;

[0064] Optionally, in step (2), the ratio of compound 2 to the 1,4-dioxane hydrochloric acid solution is 3.18 mmol: 20 mL, and optionally, the concentration of the 1,4-dioxane hydrochloric acid solution is 2 M.

[0065] Optionally, the reaction temperature is 25°C; the reaction time is 1 h;

[0066] Optionally, the solvent for the reaction includes dichloromethane;

[0067] In a specific embodiment, step (2) is described by the following formula:

[0068] ;

[0069] In a specific embodiment, a solution of 1,4-dioxane hydrochloric acid (2 M, 20 mL) was added to a solution of compound 2 (1.00 g, 3.18 mmol) in dichloromethane (10 mL), and the mixture was stirred at 25 °C for 1 h.

[0070] Optionally, a post-processing step is performed after the reaction, which includes concentrating the reaction mixture under reduced pressure and using the resulting residue directly in the next reaction without purification. This yields a colorless oily compound 3.

[0071] (3) The compound 3 was reacted with 5-bromobenzo[d]isothiazol-3-carboxylic acid to give compound 4;

[0072] Optionally, in step (3), the molar ratio of compound 3 and 5-bromobenzo[d]isothiazol-3-carboxylic acid is 1.16:0.39;

[0073] Optionally, the reaction is carried out under the conditions of benzotriazole-1-yloxy-tris(tetrahydropyrrolidinyl)phosphonium hexafluorophosphate and N,N-diisopropylethylamine; the molar ratio of compound 3 to benzotriazole-1-yloxy-tris(tetrahydropyrrolidinyl)phosphonium hexafluorophosphate is 1.16:0.465; the molar ratio of compound 3 to N,N-diisopropylethylamine is 1:1;

[0074] Optionally, the reaction temperature is 25°C; the reaction time is 1 h;

[0075] Optionally, the solvent for the reaction includes dichloromethane;

[0076] In a specific embodiment, step (3) is described by the following formula:

[0077] ;

[0078] In a specific embodiment, benzotriazole-1-yloxy-tris(tetrahydropyrrolyl)phosphonium hexafluorophosphate (PyBOP, 0.242 g, 0.465 mmol) and N,N-diisopropylethylamine (DIPEA, 0.150 g, 1.16 mmol, 0.202 mL) were added to a dichloromethane (5 mL) solution of compound 3 (0.249 g, 1.16 mmol) and 5-bromobenzo[d]isothiazolium-3-carboxylic acid (0.100 g, 0.39 mmol), and the mixture was stirred at 25 °C for 1 h.

[0079] Optionally, a post-processing is performed after the reaction, the post-processing comprising: concentrating the reaction mixture under reduced pressure, purifying the residue by silica gel column chromatography (eluent: petroleum ether solution of 35-45% ethyl acetate), monitoring by thin-layer chromatography (developing solvent: petroleum ether: ethyl acetate = 1:1, Rf = 0.6), to obtain a white solid compound 4.

[0080] (4) The compound 4 was reacted with 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole to obtain benzisothiazol carboxamide compounds;

[0081] Optionally, in step (4), the molar ratio of compound 4 and 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole is 1:1;

[0082] Optionally, the reaction is carried out in an aqueous solution of chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) and potassium carbonate; the molar ratio of compound 4 to chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) is 0.240:0.024; the concentration of the potassium carbonate aqueous solution is 2 M; and the ratio of compound 4 to the potassium carbonate aqueous solution is 0.240 mmol:0.36 mL.

[0083] Optionally, the reaction temperature is 100°C; the reaction time is 1 h; the reaction is carried out under a protective atmosphere; optionally, the protective atmosphere includes nitrogen.

[0084] Optionally, the solvent for the reaction includes dioxane;

[0085] In a specific embodiment, step (4) is described by the following formula:

[0086] ;

[0087] In a specific embodiment, a solution of compound 4 (0.110 g, 0.240 mmol) and 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1H-pyrazole (0.0500 g, 0.240 mmol) in dioxane (5 mL) was added to a solution of chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (0.0190 g, 0.024 mmol) and an aqueous solution of potassium carbonate (2 M, 0.36 mL). The mixture was stirred at 100 °C for 1 h under a nitrogen atmosphere.

[0088] Optionally, a post-processing step is performed after the reaction, which includes: concentrating the reaction mixture under reduced pressure, purifying the residue by semi-preparative reversed-phase high-performance liquid chromatography (elution gradient: 30%-60% acetonitrile in an aqueous solution containing 0.05% ammonium hydroxide for 12 min), and lyophilizing the collected solution to obtain a white solid compound of benzisothiazolium carboxamide (0.049 g, 0.11 mmol, yield 44.10%, purity 99.08%).

[0089] An inhibitor of myosin motor domain according to an embodiment of the present invention includes the benzisothiazolium carboxamide compound or its pesticide-acceptable salt as described in the embodiments of the present invention.

[0090] In this embodiment of the invention, benzisothiazolium carboxamide compounds, as inhibitors of the myosin motor domain, have the characteristics of high specificity and high activity, and can significantly inhibit the growth and development process and pathogenicity of pathogenic fungi (especially rice blast fungus).

[0091] In some embodiments, the myosin motor domain is derived from the myosin motor domain of a pathogenic microorganism; alternatively, the myosin motor domain is derived from the chitin synthase of the pathogenic microorganism, preferably the MMD of chitin synthases (Chs) of *Blastomyces oryzae*, and in a specific embodiment, the MMD of Chs5 at the N-terminus and / or the MMD of Chs6 at the N-terminus of *Blastomyces oryzae*.

[0092] In the embodiments of the present invention, benzisothiazolamide compounds, as MMD inhibitors of rice blast fungus CHS, have the characteristics of high specificity and high activity, and can significantly inhibit the growth and development process and pathogenicity of pathogenic fungi (especially rice blast fungus).

[0093] In some embodiments, the myosin motor domain inhibitor further includes a pesticide-acceptable carrier or excipient.

[0094] In some embodiments, the present invention does not impose any particular limitation on the dosage form of the myosin motor domain inhibitor, and various dosage forms may be used. For example, the dosage form may include at least one of emulsifiable concentrate, wettable powder, suspension concentrate, soluble powder, aqueous solution, fumigant, granule or seed coating agent.

[0095] The present invention relates to the application of a benzisothiazol carboxamide compound or a myosin motor domain inhibitor for the prevention and control of plant diseases caused by pathogenic microorganisms.

[0096] In the embodiments of the present invention, the benzisothiazolium carboxamide compounds and the inhibitors of myosin motor domains are characterized by high specificity and high activity, and can significantly inhibit the growth and development process and pathogenicity of pathogenic fungi (especially rice blast fungus), and can be used to prevent and control plant diseases caused by pathogenic microorganisms.

[0097] In some embodiments, the pathogenic microorganism includes pathogenic fungi, and / or, the pathogenic microorganism includes at least one of a myosin motor domain or a homologous protein thereof; optionally, the pathogenic microorganism includes pathogenic fungi containing a myosin motor domain or a homologous protein thereof; optionally, the pathogenic microorganism includes *Blastomyces oryzae*. Homologous proteins are proteins derived from different species but evolved from a common ancestral gene.

[0098] In the embodiments of the present invention, benzisothiazolium carboxamide compounds can inhibit the activity of MMD, demonstrating excellent ability to inhibit the growth and pathogenicity of pathogenic fungi (especially rice blast fungus).

[0099] In some embodiments, the plant disease includes plant fungal diseases; alternatively, the plant disease includes rice blast.

[0100] In some embodiments, it is used to inhibit the growth and development process and pathogenicity of pathogenic microorganisms.

[0101] In some embodiments, the plant includes rice, used to prevent and control rice diseases caused by pathogenic microorganisms, optionally used to prevent and control rice blast disease caused by rice blast fungus, and optionally used to inhibit the growth and development process and pathogenicity of rice blast fungus.

[0102] The present invention will now be described with reference to specific embodiments. It should be noted that these embodiments are merely descriptive and do not limit the present invention in any way.

[0103] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0104] Unless otherwise specified, the quantitative experiments in the following examples are all repeated three times, and the results are averaged.

[0105] The detection methods involved in the embodiments of this invention are industry-recognized standard procedures. For example, surface plasmon resonance (SPR) technology, with its high sensitivity and good repeatability, is widely used in the analysis of interactions between small molecules and target proteins and is one of the internationally accepted detection methods. For example, in vitro inoculation experiments using rice leaves are a standard method for detecting the effect of pesticides on the pathogenicity of rice blast fungus (People's Republic of China Agricultural Industry Standard NY / T 3257-2018).

[0106] Example 1

[0107] A benzisothiazolium carboxamide compound (A172) has the following structure:

[0108]

[0109] The preparation method of benzisothiazolium carboxamide compounds includes the following steps:

[0110]

[0111] To a solution of compound 1 (0.500 g, 1.66 mmol) and methylamine hydrochloride (0.112 g, 1.66 mmol) in dichloromethane (5 mL), benzotriazole-1-yloxy-tris(tetrahydropyrrolyl)phosphonium hexafluorophosphate (PyBOP, 1.04 g, 1.99 mmol) and N,N-diisopropylethylamine (DIPEA, 0.643 g, 4.98 mmol, 0.87 mL) were added. The mixture was stirred at 25 °C for 1 h. Liquid chromatography-mass spectrometry analysis showed a target product mass-to-charge ratio signal of 63.1% (retention time: 0.532 min). The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 50%). Thin-layer chromatography was used for monitoring (developing solvent: petroleum ether: ethyl acetate = 1:1, Rf = 0.1). A white solid compound 2 (1.000 g, crude) was obtained.

[0112]

[0113] A solution of 1,4-dioxane in hydrochloric acid (2 M, 20 mL) was added to a solution of compound 2 (1.00 g, 3.18 mmol) in dichloromethane (10 mL). The mixture was stirred at 25 °C for 1 h. Liquid chromatography-mass spectrometry analysis showed a 93.2% mass-to-charge ratio signal of the target product (retention time: 0.229 min). The reaction mixture was concentrated under reduced pressure. The resulting residue was used directly in the next reaction without purification, yielding a colorless oily compound 3 (0.550 g, crude).

[0114]

[0115] To a solution of compound 3 (0.249 g, 1.16 mmol) and 5-bromobenzo[d]isothiazolium-3-carboxylic acid (0.100 g, 0.39 mmol) in dichloromethane (5 mL), benzotriazole-1-yloxy-tris(tetrahydropyrrolyl)phosphonium hexafluorophosphate (PyBOP, 0.242 g, 0.465 mmol) and N,N-diisopropylethylamine (DIPEA, 0.150 g, 1.16 mmol, 0.202 mL) were added. The mixture was stirred at 25 °C for 1 h. Liquid chromatography-mass spectrometry analysis showed that the proportion of the target product peak was 47.8% (retention time: 0.548 min). The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether solution of 35-45% ethyl acetate). Thin-layer chromatography was used for monitoring (developing solvent: petroleum ether: ethyl acetate = 1:1, Rf = 0.6). The product was a white solid, compound 4 (0.130 g, 0.29 mmol, yield 73.86%).

[0116]

[0117] To a solution of compound 4 (0.110 g, 0.240 mmol) and 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1H-pyrazole (0.0500 g, 0.240 mmol) in dioxane (5 mL), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (0.0190 g, 0.024 mmol) and an aqueous solution of potassium carbonate (2 M, 0.36 mL) were added. The mixture was stirred at 100 °C for 1 h under a nitrogen atmosphere. Liquid chromatography-mass spectrometry analysis showed that the proportion of the target product peak was 73.9% (retention time: 0.595 min). The reaction mixture was concentrated under reduced pressure. The residue was purified by semi-preparative reversed-phase high-performance liquid chromatography (elution gradient: 30%-60% acetonitrile in an aqueous solution containing 0.05% ammonium hydroxide, for 12 min). The collected solution was lyophilized to obtain a white solid compound of benzisothiazolium carboxamide (0.049 g, 0.11 mmol, yield 44.10%, purity 99.08%).

[0118] Example 2

[0119] The interaction between the benzisothiazol carboxamide compounds described in Example 1 and the MMD protein was detected by surface plasmon resonance (SPR) method (using a Biacore 8k+ instrument).

[0120] The MMD protein of *C. oryzae* CHS was immobilized on the surface of a CM5 chip using an amino-coupling method. A series of concentration gradients (1.5625 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM) of the benzisothiazolium carboxamide compound described in Example 1 were passed through the CM5 chip surface, and changes in the reaction signal were detected. The signal was then fitted to a pre-set steady-state model, and the equilibrium dissociation constant was calculated. K D The strength of the reaction affinity K D The smaller the child, the stronger their affinity (the result is as follows). Figure 1 As shown.

[0121] from Figure 1 It can be seen that the equilibrium dissociation constants of the benzisothiazolium carboxamide compounds described in Example 1 with the MMD of rice blast fungus CHS5 and CHS6 were detected by surface plasmon resonance method. K D The concentrations were 3.51 μM and 2.19 μM, respectively, indicating that the benzisothiazol carboxamide compound described in Example 1 of this application interacts with MMD.

[0122] Example 3

[0123] The diameter of the colonies was measured after inoculating rice blast fungus P131 mycelial cakes into the center of CM medium with or without added compounds and incubating at 28°C for 7 days.

[0124] from Figure 2 It can be seen that when the CM medium contains 50 μg / mL or 100 μg / mL of the benzisothiazolium carboxamide compound described in Example 1 of this application, the radial growth of mycelium can be significantly inhibited at both concentrations. That is, the benzisothiazolium carboxamide compound described in Example 1 of this application can significantly inhibit the growth and development of rice blast fungus.

[0125] Example 4

[0126] The effect of the benzisothiazolamide compounds described in Example 1 of this application on the pathogenicity of rice blast fungus was investigated by an in vitro inoculation experiment based on spot inoculation of rice leaves.

[0127] In vitro inoculation experiments based on rice leaf spot application are the standard method for detecting the effect of pesticides on the pathogenicity of rice blast fungus (Agricultural Industry Standard of the People's Republic of China NY / T 3257-2018).

[0128] The specific steps are as follows:

[0129] (1) Cut the middle segment of the fourth leaf of a rice seedling at the four-leaf-one-heart stage, divide it into leaves about 5 cm in length, make a slit in it, and place it in a porcelain dish lined with absorbent paper. Prepare a suspension of rice blast fungus conidia and adjust its concentration to 5 × 10⁻⁶. 4 per mL.

[0130] (2) Take 10 μL of rice blast fungus spore suspension mixed with different concentrations of benzisothiazol carboxamide compounds as described in Example 1 of this application, positive control group (rice blast fungus spore suspension with 10 μg / mL pyraclostrobin added), and negative control group (rice blast fungus spore suspension without added agent) and apply it to the scratched area of ​​rice leaves. Apply 3 drops evenly to each leaf.

[0131] (3) Cultivate in the dark and moist environment in a greenhouse at 28℃ for 36 hours.

[0132] (4) Continue to culture the leaves obtained in step (3) under light conditions for 3-4 days, collect data on disease incidence and take photos for recording. The results are as follows: Figure 3 As shown; the statistical results of the lesion area are as follows. Figure 3 As shown.

[0133] from Figure 3 As can be seen from the data, the leaves treated with the benzisothiazolium carboxamide compound described in Example 1 of this application showed brown resistant lesions, just like the positive control group. However, the negative control group showed spindle-shaped lesions characteristic of rice blast. This means that the benzisothiazolium carboxamide compound described in this application can inhibit the infection of rice leaves by conidia of rice blast fungus, thus inhibiting the occurrence of rice blast and protecting the rice plants.

[0134] from Figure 3 The statistical results of the lesion area show that the inhibition of lesion area by the benzisothiazol carboxamide compound described in Example 1 of this application is concentration-dependent.

[0135] In this invention, the terms "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., refer to a specific feature, structure, material, or characteristic described in connection with that embodiment or example, which is included in at least one embodiment or example of the invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0136] Although the above embodiments have been shown and described, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Any changes, modifications, substitutions and variations made to the above embodiments by those skilled in the art are within the protection scope of the present invention.

Claims

1. A benzisothiazolium carboxamide compound, characterized in that, The structural formula of the benzisothiazol carboxamide compound is shown below: 。 2. A method for preparing the benzisothiazolium carboxamide compound according to claim 1, characterized in that, Includes the following steps: (1) Compound 1 was reacted with methylamine hydrochloride to obtain compound 2; (2) The compound 2 was mixed with a 1,4-dioxane solution of hydrochloric acid and reacted to obtain compound 3; (3) The compound 3 was reacted with 5-bromobenzo[d]isothiazol-3-carboxylic acid to give compound 4; (4) The compound 4 was reacted with 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole to obtain benzisothiazol carboxamide compounds; The structural formulas of compounds 1, 2, 3, and 4 are shown below: 。 3. The method for preparing benzisothiazolium carboxamide compounds according to claim 2, characterized in that, In step (1), the reaction is carried out under the conditions of benzotriazole-1-yloxy-tris(tetrahydropyrrolidinyl)phosphonium hexafluorophosphate and N,N-diisopropylethylamine; And / or, in step (1), the molar ratio of compound 1 and methylamine hydrochloride is 1:1; And / or, in step (1), the reaction temperature is 25°C; And / or, in step (1), the reaction time is 1 h; And / or, in step (2), the reaction temperature is 25°C; And / or, in step (2), the reaction time is 1 h.

4. The method for preparing benzisothiazolium carboxamide compounds according to claim 2, characterized in that, In step (3), the molar ratio of compound 3 and 5-bromobenzo[d]isothiazol-3-carboxylic acid is 1.16:0.39; And / or, the reaction is carried out under the conditions of benzotriazole-1-yloxy-tris(tetrahydropyrrolidinyl)phosphonium hexafluorophosphate and N,N-diisopropylethylamine; And / or, the temperature of the reaction is 25°C; And / or, the reaction time is 1 h.

5. The method for preparing the benzisothiazolium carboxamide compound according to claim 2, characterized in that, In step (4), the molar ratio of compound 4 and 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole is 1:1; And / or, the reaction is carried out in aqueous solution of chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) and potassium carbonate; And / or, the temperature of the reaction is 100°C; And / or, the reaction time is 1 h; And / or, the reaction is carried out under a protective atmosphere.

6. A myosin motor domain inhibitor, characterized in that, Includes the benzisothiazolium carboxamide compounds of claim 1 or their pesticide-acceptable salts.

7. The myosin motor domain inhibitor according to claim 6, characterized in that, This also includes pesticide-acceptable carriers or excipients; And / or, the dosage form of the myosin motor domain inhibitor is selected from emulsifiable concentrates, wettable powders, suspension concentrates, soluble powders, aqueous solutions, fumigants, granules, or seed coatings; And / or, the myosin motor domain is derived from chitin synthase of pathogenic microorganisms.

8. The application of a benzisothiazolium carboxamide compound according to claim 1 or a myosin motor domain inhibitor according to claim 6 or 7, characterized in that, Used to prevent and control plant diseases caused by pathogenic microorganisms.

9. The application according to claim 8, characterized in that, The pathogenic microorganisms include pathogenic fungi; And / or, the pathogenic microorganism includes at least one of the myosin motor domain or its homologous protein; And / or, the plant diseases include plant fungal diseases.

10. The application according to claim 9, characterized in that, The pathogenic microorganisms include rice blast fungus; And / or, the plant disease includes rice blast; And / or, the plant includes rice.

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