Application of nitrostyrene compound in prevention and / or control of pathogenic microorganisms and medicine

Abstract

The invention discloses application of a nitrostyrolene compound in prevention and / or control of pathogenic microorganisms and a medicine, further discloses a pharmaceutical preparation containing the nitrostyrolene compound, and relates to the technical field of pharmaceutical chemistry. The compound has a good application prospect in the aspects of preventing and treating agricultural pathogenic fungi, agricultural pathogenic bacteria and agricultural pathogenic nematodes, and is expected to be developed into a novel antibacterial or nematode-killing active ingredient of a medicinal preparation.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to the use of a nitrostyrene compound in the prevention and / or control of pathogenic microorganisms, and more specifically to the application and drug of a nitrostyrene compound in the prevention and / or control of pathogenic microorganisms. Background Technology

[0002] Pathogenic microorganisms, a vast group of tiny organisms found in nature, encompass various types including fungi, bacteria, nematodes, and viruses. Their invasion of core areas of human society has never ceased, posing a persistent and serious cross-sectoral threat to key industries such as agricultural production and healthcare. In agriculture, diseases caused by various pathogenic microorganisms have become one of the core bottlenecks restricting global food security and the improvement of agricultural product quality.

[0003] Among these, fungal diseases, such as damping-off, can infect various crops including wheat, cotton, and vegetables, causing seedlings to wither and die, resulting in a significant decrease in field emergence rates. Sclerotinia rot in rapeseed, a typical fungal disease, spreads during the flowering and pod-setting stage, damaging stems and pods, and in severe cases can reduce rapeseed yield by more than 30%, also affecting the quality of rapeseed oil. Bacterial leaf blight in rice, caused by Xanthomonas, spreads through wind, rain, and agricultural operations. After infection, rice leaves develop white spots, hindering photosynthesis and directly leading to an increase in empty grains. Furthermore, nematode diseases are equally significant. Root-knot nematodes and other species can parasitize crop roots, forming root knots and disrupting nutrient absorption pathways, weakening crop growth, reducing resistance, and further exacerbating the risk of yield reduction. Outbreaks of these diseases not only affect farmers' income but also pose a challenge to the stability of the global food supply chain. Therefore, it is evident that scientific, effective, and precise prevention and control of various pathogenic microorganisms is the foundation for ensuring a high-quality and safe supply of agricultural products and stabilizing agricultural production order. Currently, chemical fungicides, nematicides, and various antimicrobial drugs remain the mainstream means of controlling pathogenic microorganisms globally due to their direct and rapid effects, playing an irreplaceable role in agricultural pest and disease control and clinical infection treatment. However, for a long time, both the blind increase in pesticide dosage and extension of treatment cycles in pursuit of high yields in agricultural production and the irrational use of antimicrobial drugs in clinical medicine have led to a common problem—the continuous enhancement of drug resistance in pathogenic microorganisms.

[0004] Mechanistically, the selective pressure of a single drug promotes the survival and proliferation of drug-resistant mutant strains within a pathogenic microbial population. Simultaneously, the transmission of resistance genes between different strains further expands the resistant population, ultimately leading to a gradual decline in the effectiveness of drug control and forcing continuous increases in drug dosage, creating a vicious cycle of "drug use - resistance - increased dosage - more severe resistance." This cycle not only causes a sustained increase in the infection rate and severity of pathogenic microorganisms in agricultural production but also increases production costs and environmental pressure. Furthermore, it leads to difficulties in the treatment of infectious diseases in the medical field, further exacerbating public health risks. Therefore, in order to overcome the limitations of traditional drugs, researching and developing new, efficient, safe, and low-drug-resistance bactericides / nematicides and antimicrobial drugs has become the core direction and urgent need of scientific research in fields such as agricultural plant protection and pharmaceutical research and development. Summary of the Invention

[0005] To address the above shortcomings, this invention provides an application and drug for nitrostyrene compounds in the prevention and / or control of agricultural pathogenic fungi, agricultural pathogenic bacteria, and agricultural pathogenic nematodes. This solves the problems of existing antimicrobial agents, such as high dosage, poor control efficacy, and easy development of drug resistance. It can be used to control various diseases caused by the above pathogenic microorganisms. The specific technical solution is as follows: The use of a nitrostyrene compound in the prevention and / or control of pathogenic microorganisms, including fungi, bacteria, nematodes and / or viruses.

[0006] Preferably, the structure of the nitrostyrene compound is selected from: Preferably, the structure of the nitrostyrene compound is selected from: .

[0007] Preferably, the structure of the nitrostyrene compound is selected from: .

[0008] Preferably, the structure of the nitrostyrene compound is selected from: .

[0009] Preferably, the pathogenic microorganisms include agricultural pathogenic fungi, agricultural pathogenic bacteria, agricultural pathogenic nematodes, and / or human-derived pathogens.

[0010] Preferably, the agricultural pathogenic fungus includes Rhizoctonia solani. Rhizoctonia solani Sclerotium sclerotiorum Sclerotinia sclerotiorum Botrytis cinerea Botrytis cinerea Fusarium graminearum Fusarium grasses Rice blast fungus Magnaporthe rice and Fusarium pseudograss Fusarium pseudograsses The agricultural fungal disease mentioned is Rhizoctonia solani. Rhizoctonia solani Sclerotium sclerotiorum Sclerotinia sclerotiorum Botrytis cinerea Botrytis cinerea Fusarium graminearum Fusarium grasses Rice blast fungus Magnaporthe rice In addition, Fusarium graminearum causes damping-off, sclerotinia rot in rapeseed, gray mold in tomato, scab in wheat, rice blast, and wheat stem rot.

[0011] Preferably, the agricultural pathogenic bacteria include *Bacillus thuringiensis*, the pathogen that causes bacterial blight in rice. Xanthomonas oryzae PXO99A, Citrus canker pathogen Xanthomonas axonopodis jx-6 and Ralstonia solanacearum Pseudomonas solanacearum The bacterial disease of the plant is *Bacterium tumefaciens*, the pathogen causing bacterial blight of rice. Xanthomonas oryzae PXO99A, Citrus canker pathogen Xanthomonas axonopodis jx-6 and Ralstonia solanacearum Pseudomonas solanacearum These diseases respectively cause bacterial blight in rice, citrus canker, and bacterial wilt.

[0012] Preferably, the agricultural pathogenic nematode is the southern root-knot nematode. Meloidogyne incongruus pine wood nematode Busaphelenchus xylophilus At least one of the following, the agricultural pathogenic nematode disease being the southern root-knot nematode. Meloidogyne incongruus pine wood nematode Busaphelenchus xylophilus The diseases caused by plant root-knot nematodes and pine wilt nematodes, respectively; the human-borne pathogens include Escherichia coli. Escherichia coli ATCC 25922, Staphylococcus aureus Staphylococcus aureus Newman Candida albicans Candida albicans ATCC 24433.

[0013] Preferably, the use of a nitrostyrene compound as the sole active ingredient in a drug for the prevention and / or control of pathogenic microorganisms.

[0014] Preferably, a pharmaceutical preparation for the prevention and / or control of pathogenic microorganisms includes the nitrostyrene compounds described above.

[0015] Preferably, the nitrostyrene compound is the sole active ingredient in the pharmaceutical preparation, and its weight percentage is 1-99%.

[0016] Preferably, the pharmaceutical preparation further includes pharmaceutically acceptable adjuvants.

[0017] Preferably, the additives are selected from one or more of the following: dispersants, wetting agents, fillers, emulsifiers, thickeners, dispersion media, disintegrants, solvents, preservatives, antifreeze agents, specific gravity regulators, defoamers, oil-phase wall materials, aqueous-phase wall materials, and water.

[0018] Preferably, the dosage form of the pharmaceutical preparation is one of granules, dry suspensions, aqueous suspensions, dispersible oil suspensions, microcapsule suspensions, wettable powders, emulsifiable concentrates, water emulsions, water-dispersible granules, and seed treatment agents.

[0019] The present invention achieves at least the following beneficial effects: 1. The nitrostyrene compounds discovered in this invention exhibit good antibacterial or nematicidal activity against 6 types of agricultural pathogenic fungi, 3 types of agricultural pathogenic bacteria, 3 types of human pathogenic bacteria, and 2 types of agricultural pathogenic nematodes, and can be further developed as lead compounds for antibacterial and nematicidal purposes.

[0020] 2. The nitrostyrene compounds discovered in this invention are easy to synthesize, have excellent activity, and a broad spectrum of resistance against pathogenic microorganisms. They exhibit excellent antibacterial and nematicidal activity against a variety of agricultural pathogenic microorganisms and have good application prospects in the control of agricultural pathogenic fungi, agricultural pathogenic bacteria, and agricultural pathogenic nematodes. They can be applied to the development of novel high-activity antibacterial and antinematic drugs and are expected to be developed into active ingredients for novel antibacterial or nematicidal drug formulations.

[0021] 3. This invention provides nitrostyrene compounds and drugs derived therefrom that can be used to prevent and control various diseases in agriculture caused by pathogenic fungi, agricultural pathogenic bacteria, agricultural pathogenic nematodes, and other pathogenic microorganisms. These compounds have good antibacterial and nematicidal activities, which help ensure the quality of agricultural products and human health and safety. Attached Figure Description

[0022] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 A diagram showing the current state of crops after the second application of this invention and / or a suspension containing 35% of the X-17 compound of this invention; Figure 2 This is a diagram showing the current state of crops that have not been sprayed with the drug formulation of this invention during the same growth stage. Detailed Implementation

[0024] The specific embodiments of the present invention are described in detail below, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments. Unless otherwise defined, all technical terms used below have the same meaning as commonly understood by those skilled in the art. The technical terms used herein are for the purpose of describing specific embodiments only and are not intended to limit the scope of protection of the present invention. Unless otherwise specifically stated, all raw materials, reagents, instruments, and equipment used in the present invention are commercially available or can be prepared by existing methods.

[0025] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0026] Based on the aforementioned challenges and to overcome the limitations of traditional drugs, the inventors of this application have continuously explored and researched in the field of chemical drugs, aiming to develop novel drugs that address multiple objectives: In terms of efficacy, they need to possess broader-spectrum antibacterial and bactericidal activity to effectively combat existing drug-resistant strains and complex infections by multiple pathogenic microorganisms; in terms of safety, agricultural drugs need to reduce pollution to soil, water sources, and beneficial organisms, meeting the needs of green agriculture development, while medical drugs need to minimize damage to the normal human flora and toxic side effects; regarding the risk of drug resistance, they need to delay the development of drug resistance in pathogenic microorganisms through innovative target mechanisms, optimized drug structures, or the development of compound formulations. Simultaneously, the promotion and application of new control technologies and drugs must be coupled with scientific medication guidance and the construction of a drug resistance monitoring system to form a complete closed loop of "research and development-application-control," in order to fundamentally solve the problem of pathogenic microorganism control and provide solid support for agricultural safety and human health.

[0027] During the research and development process, the inventors discovered that nitrostyrene compounds and their derivatives possess a wide range of biological activities, such as antibacterial, antiparasitic, and anticancer effects. Nitrostyrene compounds and their derivatives are currently widely used in chemical synthesis and drug development. Derivatization and structural modification of the nitrostyrene core can further expand their applications and significantly enhance their biological activity. However, existing nitrostyrene compounds and their derivatives have relatively complex structures and complex synthesis processes, and their applicability is limited; some can only kill fungi, while others can only kill bacteria, failing to achieve broad-spectrum sterilization.

[0028] After testing the anti-agricultural pathogenic fungi, agricultural pathogenic bacteria, and agricultural pathogenic nematodes of the nitrostyrene derivatives of the present invention, the results showed that these compounds have broad-spectrum and excellent antibacterial and nematicidal activities, and some compounds have better activity than the positive control drug. Finally, the technical solution of the present invention was developed.

[0029] Example 1: Determination and Results of the Anti-Agricultural Pathogenic Fungi Activity of Nitrostyrene Compounds (1) Test reagent: Nitrostyrene compounds.

[0030] (2) Test strain: Rhizoctonia solani Rhizoctonia solani Sclerotium sclerotiorum Sclerotinia sclerotia Botrytis cinerea Botrytis cinerea Fusarium graminearum Fusarium gramineae Rice blast fungus Magnaporthe rice and Fusarium pseudograss Fusarium pseudogramineum The agricultural pathogenic fungi used in this experiment were strains preserved in the laboratory at 4℃.

[0031] (3) Test method: The mycelial growth rate method was used.

[0032] In this experiment, all plant pathogenic fungi were treated with potato agar-dextrose medium (PDA). The PDA medium formula was: 200g potato (peeled), 20g glucose, 15g agar, and 1000mL deionized water. The medium was then autoclaved at 121℃ for 20 minutes and then used for later use.

[0033] Activation method of strain: Incubate agricultural pathogenic fungi on PDA plates at 25°C for 3-6 days.

[0034] Preparation of drug-coated plates: Pour PDA culture medium sterilized at 121℃ into a petri dish, cool to 45-50℃, and add different concentrations of the test compound to prepare drug-coated plates.

[0035] Inoculation and cultivation: In a clean bench, use a punch to make a 5mm diameter mycelium cake at the edge of the mycelium after 3-6 days of cultivation (with the growth conditions as uniform as possible), then use an inoculation needle to pick it up and transfer it to the center of the substrate, and then incubate it upside down in an incubator (25℃).

[0036] Results determination: After the hyphae in the blank control group were fully grown, the growth diameter of the hyphae in the drug-treated group was measured by the cross-cross method, and the growth inhibition rate of this type of compound on hyphae was calculated.

[0037] Inhibition rate (%) = (control mycelium diameter - treated mycelium diameter) / (control mycelium diameter - mycelium cake diameter) × 100. Three parallel experiments were set up for each concentration to determine the inhibition rate of this type of compound. The results are shown in Table 1.

[0038] Table 1. Inhibition rate of nitrostyrene compounds against agricultural pathogenic fungi at 50 μg / mL Note: "-" indicates that the inhibition rate of the compound was not determined.

[0039] As shown in Table 1, the nitrostyrene compounds involved in this invention are effective against Rhizoctonia solani. Rhizoctonia solani Sclerotium sclerotiorum Sclerotinia sclerotiorum Botrytis cinerea Botrytis cinerea Fusarium graminearum Fusarium gramineae Rice blast fungus Magnaporthe rice and Fusarium pseudograss Fusarium pseudogramineum All six plant pathogenic fungi exhibited varying degrees of inhibitory activity, showing high activity against the tested pathogenic fungi and being superior to or comparable to the positive control. Furthermore, some compounds exhibited broad-spectrum activity. Therefore, the nitrostyrene compounds of this invention have value for further research and development in the area of ​​resistance to agricultural pathogenic fungi.

[0040] Example 2: Determination and Results of the Anti-Agricultural Pathogenic Bacteria Activity of Nitrostyrene Compounds (1) Test reagent: Nitrostyrene compounds.

[0041] (2) Test strain: Rice bacterial blight pathogen Xanthomonas oryzae PXO99A, Citrus canker pathogen Xanthomonas axonopodis jx-6 and Ralstonia solanacearum Pseudomonas solanacearum The strains used in this experiment were strains that were frozen in the laboratory at -80℃ with 30% glycerol.

[0042] (3) Test method: The micro-broth dilution method was used. The frozen strains were taken out and streaked onto NB solid medium (beef extract: 3g, peptone: 5g, yeast powder: 1g, sucrose: 10g, agar: 15g, distilled water: 1L, pH 7.0; sterilized at 121℃ for 20min), and incubated at 28℃ until single colonies appeared. Single colonies from the solid medium were transferred to NB liquid medium (beef extract: 3g, peptone: 5g, yeast powder: 1g, sucrose: 10g, distilled water: 1L; sterilized at 121℃ for 20min), and cultured on a shaker at 28℃ and 180rpm until the logarithmic growth phase (OD value between 0.3 and 0.6). The strains in the logarithmic growth phase were diluted with NB liquid medium to approximately 10... 6CFU / mL is prepared for use. Dissolve the compounds separately in DMSO, add them to the liquid culture medium, mix thoroughly, and prepare a drug-containing liquid culture medium with a concentration of 200 μg / mL. Take 50 μL of the drug-containing culture medium and the same volume of approximately 10... 6 CFU / mL bacterial culture was added to the wells of a 96-well plate, resulting in a final drug concentration of 100 μg / mL. A control of 100 μL of bacterial culture containing an equal amount of DMSO was used. The 96-well plates were incubated at 28°C for 24–48 h until bacterial growth was observed in the control group (OD value between 0.25 and 0.35). The OD value of the bacterial culture in each well was measured using a microplate reader. 600 In addition, the OD values ​​of 100 μL of liquid culture medium and a 100 μg / mL drug concentration were measured to correct for the OD values ​​caused by the culture medium and the drug itself. The formulas for calculating the corrected OD value and inhibition rate are as follows: Correcting OD 600 =O2 in bacterial culture medium 600 - OD of sterile cultures 600 ; All experiments were conducted in triplicate to determine the inhibitory activity of nitrostyrene compounds against the three tested plant pathogenic bacteria. The results are shown in Table 2.

[0043] Table 2. Inhibition rate (%) of nitrostyrene compounds against plant pathogenic bacteria at 100 μg / mL. Note: "-" indicates that the inhibition rate of the compound was not determined.

[0044] The drug-containing liquid culture medium was diluted in a 96-well plate by a twofold dilution method to obtain a series of 50 μL drug-containing culture media of different concentrations. Then, the inhibition rate corresponding to the series of concentrations was determined according to the above experimental method. The lowest concentration with an inhibition rate greater than 90% was defined as MIC (μg / mL), and the MIC activity was measured.

[0045] As shown in Table 2, the nitrostyrene compounds provided by this invention are effective against the tested rice bacterial blight pathogen. Xanthomonas oryzae PXO99A, Citrus canker pathogen Xanthomonas axonopodis jx-6 and Ralstonia solanacearum Pseudomonas solanacearumThese three plant bacteria exhibit excellent antibacterial activity, and most of the compounds showed better MIC values ​​than the positive control drug quinolinone in the assays for agricultural bacteria. In particular, the highly active compound X-10 showed MIC values ​​of 3.12 μg / mL, 1.56 μg / mL, and 3.12 μg / mL against rice bacterial blight, citrus canker, and Ralstonia solanacearum, respectively. Its inhibitory activity was significantly better than that of the control drug quinolinone, making it worthy of further development as a fungicide.

[0046] Example 3: Determination of the toxic activity of nitrostyrene compounds against pine wood nematodes (1) Obtaining pine wood nematodes: The pine wood nematodes used in this experiment were obtained by laboratory culture using potato agar-glucose medium (PDA). PDA medium formula: 200g potato (peeled), 20g glucose, 15g agar, 1000mL deionized water. After autoclaving at 121℃ for 20min, the medium was taken out for use. Botrytis cinerea was inoculated onto the PDA medium, and the pine wood nematodes on the Botrytis cinerea were collected using the Bellman funnel method for experimental use.

[0047] (2) Sample preparation: The nitrostyrene compounds used in this example were synthesized in the laboratory.

[0048] (3) Experimental methods: The nematicidal activity of the compounds was determined by the immersion method. Sterile water and treatment liquid systems prepared by different concentrations of the test compound or control drug were added to 48-well plates. The number of nematodes per well was controlled to be about 50. DMSO (dimethyl sulfoxide) of equal concentration was used as a control. Each experimental group was set up with three replicates. The nematodes were incubated at 25°C and observed under a stereomicroscope after 24 hours. Nematodes that could move freely or respond to the shaking of the 48-well plate by moving their heads and tails were identified as live nematodes. After calculating the corrected mortality rate corresponding to each concentration, the median lethal concentration (LC50) of the highly toxic nematicidal compound was determined by IBM statistics SPSS27.0 statistical analysis software.

[0049] (4) Experimental Results The toxic activity of nitrostyrene derivatives against pine wood nematode is shown in Table 3.

[0050] Table 3. Toxicity and LC50 values ​​of 50 μg / mL nitrostyrene compounds against pine wood nematode. Note: "-" indicates that the value of this compound was not determined.

[0051] As shown in Table 3, some of the nitrostyrene compounds involved in this invention have good nematicidal activity against pine wood nematodes, and their inhibition rate is better than or comparable to that of the positive control drug fluopyram.

[0052] Example 4: Assay of the toxic activity of nitrostyrene compounds against second-instar larvae of the southern root-knot nematode (1) Obtaining second-instar larvae of Southern Root-Knot Nematode: Take cucumber roots infected with Southern Root-Knot Nematode, wash off the soil, cut the root knots into pieces, prepare a 1% sodium hypochlorite solution to wash the root knots thoroughly for 1 min, wash off the remaining sodium hypochlorite solution with sterile water, collect the washed nematode eggs with a 500-mesh sieve, place the nematode eggs in a petri dish with sterile water, incubate in a constant temperature incubator at 28℃ for 4 days, and collect the hatched second-instar larvae of Southern Root-Knot Nematode as experimental insects.

[0053] (2) Sample preparation: The nitrostyrene compounds used in this example were synthesized in the laboratory, as in Example 3.

[0054] (3) Experimental method: The method is the same as in Example 3.

[0055] (4) Experimental results: The toxic activity of nitrostyrene derivatives against second-instar larvae of southern root-knot nematodes is shown in Table 4.

[0056] Table 4. Toxicity and LC50 values ​​of 50 μg / mL nitrostylenoids against second-instar larvae of *Sinapis alba*. Note: "-" indicates that the value of this compound was not determined.

[0057] As shown in Table 4, some of the nitrostyrene compounds involved in this invention have good nematicidal activity against second-instar larvae of the southern root-knot nematode, and their inhibition rate is better than or comparable to that of the positive control drug fluopyram.

[0058] The preparation processes for various formulations of the pesticides of this invention are as follows: The preparation method of suspension pesticide is as follows: the raw materials are sheared at high speed in proportion, and then added to a sand mill for grinding for 2-3 hours to obtain suspension pesticide formulation.

[0059] The preparation method of water-dispersible granule pesticide is as follows: the raw materials are pulverized by air jet milling, granulation, drying and sieving in proportion to prepare water-dispersible granule pesticide.

[0060] The preparation method of wettable powder pesticides is as follows: the raw materials are air-jet pulverized and sieved in proportion to prepare wettable powder pesticide formulation.

[0061] The preparation method of emulsifiable concentrate pesticide is as follows: dissolve the active ingredient in a solvent, add other components and stir to mix evenly to obtain the emulsifiable concentrate pesticide.

[0062] Example 5: 35% Compound X-17 Suspension A pharmaceutical suspension for the prevention and / or control of pathogenic microorganisms comprises the following raw materials in weight percentage: 35% compound X-17, 4% alkyl polyoxyethylene ether (dispersant), 1% alkyl alcohol ether phosphate (wetting agent), 4% sodium dodecyl sulfate (dispersant), 0.03% xanthan gum (thickener), 0.02% xanthan gum (thickener), 0.02% ethylene glycol (antifreeze agent), and water to make up to 100%.

[0063] Example 6: 40% Compound X-98 Suspension A pharmaceutical suspension for the prevention and / or control of pathogenic microorganisms comprises the following raw materials in weight percentage: 40% compound X-98, 2% alkyl naphthalene sulfonate formaldehyde condensate (dispersant), 2% alkyl alcohol ether phosphate (wetting agent), 3% sodium dodecyl sulfate (dispersant), 0.02% xanthan gum (thickener), 0.03% ethylene glycol (antifreeze agent), and water to make up to 100%.

[0064] Example 7: 50% Compound X-100 Water Dispersible Granules A water-dispersible granule formulation for the prevention and / or control of pathogenic microorganisms comprises the following raw materials in weight percentage: 50% compound X-100, 3.5% alkyl naphthalene sulfonate formaldehyde condensate (dispersant), 1.7% lignin sulfonate (dispersant), 1.5% separating powder (wetting agent), 1% sodium dodecyl sulfate (dispersant), and kaolin to make up 100%.

[0065] Example 8: 30% Compound X-106 Water Dispersible Granules A water-dispersible granule formulation for the prevention and / or control of pathogenic microorganisms comprises the following raw materials in weight percentage: 30% compound X-106, 3% alkyl naphthalene sulfonate formaldehyde condensate (dispersant), 2.5% lignin sulfonate (dispersant), 1% separating powder (wetting agent), 1% sodium dodecyl sulfate (dispersant), and kaolin to make up 100%.

[0066] Example 9: 60% Compound X-14 Wettable Powder A wettable powder for the prevention and / or control of pathogenic microorganisms, comprising the following raw materials in weight percentage: 60% compound X-14, 4.5% alkyl naphthalene sulfonate formaldehyde condensate (dispersant), 4% lignin sulfonate (dispersant), 2.5% wetting agent, and kaolin to make up 100%.

[0067] Example 10: 40% Compound ZY-6 Wettable Powder A wettable powder for the prevention and / or control of pathogenic microorganisms comprises the following raw materials by weight percentage: 40% compound ZY-6, 7% lignin sulfonate (dispersant), 1.6% sodium methylene bis(naphthalene) sulfonate (wetting agent), and kaolin to make up 100%.

[0068] Example 11: 20% Compound X-5 Wettable Powder A wettable powder for the prevention and / or control of pathogenic microorganisms, comprising the following raw materials by weight percentage: 20% compound X-5, 6% high molecular weight polycarboxylate (dispersant), 3% dodecyl polyoxyethylene ether phosphate (wetting agent), and kaolin to make up 100%.

[0069] Example 12: 25% Compound X-17 Dry Suspension A pharmaceutical dry suspension for the prevention and / or control of pathogenic microorganisms comprises the following raw materials in weight percentage: 25% compound X-17, 6.4% lignin sulfonate, 3.2% polycarboxylate, 3% polyalkylene oxide modified silane, 0.05% pH adjuster, and starch to 100%.

[0070] The preparation process of drug dry suspension includes the following steps: S1. Mix the compound X-17 technical material and various adjuvants in proportion to obtain a solid material, add water, and shear at high speed to obtain a mixed slurry. S2. The mixed slurry from step S1 is milled to obtain a suspension. S3: Spray dry the suspension of S2 to finally prepare a dry suspension.

[0071] Example 13: 40% Compound X-106 Emulsifiable Oil An emulsifiable concentrate for the prevention and / or control of pathogenic microorganisms comprises the following raw materials in weight percentage: 40% compound X-106, 5.4% alkylphenol formaldehyde resin polyoxyethylene ether, 3.6% agricultural emulsion 500#, 20% xylene, and methyl oleate to make up 100%.

[0072] Field efficacy trial for controlling rice sheath blight Test reagents: Pharmaceutical preparations obtained in Examples 5 to 11 Control reagent: 40% difenoconazole suspension, registration certificate number PD20132295, Shaanxi Thompson Biotechnology Co., Ltd.

[0073] Experimental Methods: Following the guidelines in GB / T17980.20-2000 "Field Efficacy Test Guidelines for Pesticides (I) - Control of Rice Sheath Blight with Fungicides", a 3WBD-20L backpack manual sprayer was used to apply the pesticide before the onset of rice sheath blight, ensuring even spraying. In rice paddies in Santang Village, Xinwei Town, Binyang County, Nanning City, Guangxi Province, the first spray was conducted on October 9, 2025, and the second spray was conducted on September 15, 2025, for a total of two applications. Efficacy surveys were conducted 7 and 14 days after the second application, using a diagonal sampling method. Five sampling points were taken from each plot, and five adjacent clumps were surveyed at each point, for a total of 25 clumps. The total number of plants and the number of diseased plants were recorded. Each treatment was replicated four times, and the average value was taken. The disease index and control effect were calculated. The results are shown in Table 5.

[0074] ; .

[0075] Table 5. Results of field efficacy trials for controlling rice sheath blight. As shown in Table 5 above, the nitrostyrene compounds of this invention have better efficacy in field trials for controlling rice sheath blight than the currently available commercially available 40% difenoconazole suspension for controlling rice sheath blight.

[0076] In summary, the nitrostyrene compounds of this invention are effective against Rhizoctonia solani. Rhizoctonia nightshade Sclerotium sclerotiorum Sclerotinia sclerotiorum Botrytis cinerea Botrytis cinerea Fusarium graminearum Fusarium gramineae Rice blast fungus Magnaporthe rice and Fusarium pseudograss Fusarium pseudograsses The fungi that cause damping-off, sclerotinia stem rot in rapeseed, gray mold in tomato, scab in wheat, rice blast, and wheat stem rot are all susceptible to bacterial blight of rice. Xanthomonas oryzae PXO99A, Citrus canker pathogen Xanthomonas axonopodis jx-6 and Ralstonia solanacearum Pseudomonas solanacearum The bacteria that cause rice bacterial blight, citrus canker, and bacterial wilt are three agricultural pathogens, and are effective against Escherichia coli. Escherichia coli ATCC25922, Staphylococcus aureus Staphylococcus aureus Newman and Candida albicans Candida albicans Human-borne bacterial diseases caused by three human-borne bacteria, ATCC 24433, and the southern root-knot nematode. Meloidogyne incongnita pine wood nematode Busaphelenchus xylophilusThe two agricultural pathogenic nematodes that cause plant root-knot nematode disease and pine wilt disease have shown good antibacterial and nematicidal activity, and have further research and development value in the prevention and control of pathogenic microorganisms.

[0077] The foregoing description of specific exemplary embodiments of the invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the invention, as well as various different choices and variations. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. The application of a nitrostyrene compound in the prevention and / or control of pathogenic microorganisms, characterized in that, The structures of the nitrostyrene compounds are selected from: 。 2. The application according to claim 1, characterized in that, The structures of the nitrostyrene compounds are selected from: 。 3. The application according to claim 1, characterized in that, The pathogenic microorganisms include agricultural pathogenic fungi, agricultural pathogenic bacteria, and agricultural pathogenic nematodes.

4. The application according to claim 3, characterized in that, The agricultural pathogenic fungi include Rhizoctonia solani. Rhizoctonia solani Sclerotium sclerotiorum Sclerotinia sclerotiorum Botrytis cinerea Botrytis cinerea Fusarium graminearum Fusarium graminearum Rice blast fungus Magnaporthe oryzae and Fusarium pseudograss Fusarium pseudograminearum .

5. The application according to claim 3, characterized in that, The agricultural pathogens include rice bacterial blight pathogens. Xanthomonas oryzae PXO99A, Citrus canker pathogen Xanthomonas axonopodis jx-6 and Ralstonia solanacearum Pseudomonas solanacearum .

6. The application according to claim 3, characterized in that, The agricultural pathogenic nematode is *Strombus heterophylla*. Meloidogyne incongnita pine wood nematode Busaphelenchus xylophilus At least one of them.

7. A pharmaceutical preparation for the prevention and / or control of pathogenic microorganisms, characterized in that: The pharmaceutical preparation includes the nitrostyrene compounds as described in claims 1 to 6.

8. The pharmaceutical preparation according to claim 7, characterized in that, In the pharmaceutical preparation, nitrostyrene compounds are the sole active ingredient, and their weight percentage ranges from 1% to 99%.

9. The pharmaceutical preparation according to claim 7, characterized in that, The pharmaceutical preparation further includes pharmaceutically acceptable adjuvants selected from one or more of the following: dispersants, wetting agents, fillers, emulsifiers, thickeners, dispersion media, disintegrants, solvents, preservatives, antifreeze agents, specific gravity regulators, defoamers, oil-phase wall materials, aqueous-phase wall materials, and water.

10. The pharmaceutical preparation according to claim 7, characterized in that, The dosage form of the pharmaceutical preparation is one of the following: granules, dry suspension, aqueous suspension, dispersible oil suspension, microcapsule suspension, wettable powder, emulsifiable concentrate, water emulsion, water-dispersible granules, and seed treatment agent.

Images