Oxidic compounds containing cinnamoyl-substituted unsaturated nitrogen-containing heterocycles, compositions, methods of preparation and use

Abstract

The application provides a cinnamoyl-substituted unsaturated nitrogen-containing heterocyclic oxygen-containing compound, a composition, a preparation method and application, and belongs to the technical field of medicinal chemistry.The structural formula of the compound provided by the application is The application takes a cinnamic acid structure as a lead compound, then takes a nitrogen-containing heterocyclic ring such as piperazine, piperidine and homopiperazine as a connecting group, and on this basis, different pharmacophores are connected to synthesize a series of cinnamoyl-substituted unsaturated nitrogen-containing heterocyclic oxygen-containing compounds.The compounds have excellent inhibitory effect on plant pathogenic bacteria such as rice bacterial leaf blight bacteria, citrus canker bacteria and kiwi canker bacteria.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical chemistry, in particular to an oxygen-containing compound containing a cinnamoyl-substituted unsaturated nitrogen-containing heterocycle, a composition, a preparation method and applications. BACKGROUND

[0002] Plant bacterial diseases are a kind of plant diseases caused by plant pathogenic bacteria, which are extremely harmful, and have the characteristics of outbreak, prevalence and destructiveness. Once infected, the entire planting area may become a lifelong epidemic area. In current agricultural production, common plant bacterial diseases include bacterial leaf streak of rice, bacterial leaf blight of rice, citrus canker, kiwi canker, Chinese cabbage soft rot, tobacco bacterial wilt and tomato canker, etc. These diseases have a serious impact on the yield and quality of host plants. Long-term practice has proved that chemical control is the most effective means to control various plant bacterial diseases. However, the excessive and blind use of pesticides not only increases production costs and affects the ecological environment, but also induces pathogenic bacteria to develop drug resistance, resulting in poor control effect of traditional pesticides. Therefore, it is urgent to find new pesticides with novel mechanism of action, green and high efficiency, and low toxicity, in order to enhance the effective control of plant bacterial diseases and provide strong guarantee for the green development of agricultural production.

[0003] From a historical perspective, natural products have always been a valuable source of antibacterial drugs. Although the development of more convenient artificial synthesis methods is rapid, there are still certain limitations in structural diversity, etc. The rich drug structures and active groups contained in natural products provide rich chemical diversity. The low toxicity and environmental friendliness of natural products are also advantages that cannot be ignored. With the continuous maturity of natural product separation, structure analysis and biosynthesis technologies, more attention has been paid to the research and application of natural products. Natural product derivatization and synthetic drugs based on natural products have become an effective means to discover new antibacterial pesticides. Cinnamic acid is an aromatic compound extracted from the stems of Cinnamomum cassia. It and its derivatives exhibit a variety of biological activities and are widely used in traditional medicine and pharmaceutical preparations for the treatment of gastritis, colds, platelet aggregation inhibition and sterilization. Currently, in the field of pesticide research, it has been found that derivatives of cinnamic acid have significant antibacterial, antiviral and herbicidal potential. With the continuous progress of science and technology, in-depth research on cinnamic acid and its derivatives will provide important support for the discovery of new antibacterial pesticides.

[0004] At present, there are a large number of studies on the biological activity of cinnamic acid compounds and the biological activity of nitrogen-containing heterocyclic compounds. Therefore, how to continue to use cinnamic acid structure as a lead compound and use piperazine, piperidine and homopiperazine as linking groups to modify and find active target compounds with high bactericidal efficiency is a problem that needs to be solved at present. SUMMARY

[0005] In view of the above, the present application aims to provide a cinnamoyl-substituted unsaturated nitrogen-containing heterocyclic oxygen-containing compound, a composition, a preparation method and an application, and the cinnamoyl-substituted unsaturated nitrogen-containing heterocyclic oxygen-containing compound provided by the present application has excellent inhibitory effect on plant pathogenic bacteria.

[0006] In order to achieve the above-mentioned purpose, the present application provides the following technical solutions: a cinnamoyl-substituted unsaturated nitrogen-containing heterocyclic oxygen-containing compound having the structure of formula I or a stereoisomer thereof, or a salt thereof or a solvate thereof:

[0007]

[0008] wherein R1 and R3 are each independently selected from one or more of hydrogen, deuterium, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl, optionally substituted or unsubstituted alkoxy, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted aryl, and optionally substituted or unsubstituted heteroaryl;

[0009] R2 is selected from hydroxyl or hydrogen;

[0010] X is selected from one or more of amino, optionally substituted or unsubstituted aminopiperidine, optionally substituted or unsubstituted piperazine, optionally substituted or unsubstituted homopiperazine, and optionally substituted or unsubstituted nitrogen-containing heterocycle.

[0011] Preferably, R1 and R3 are each independently selected from one or more of hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, propenyl, allyl, methoxy, ethoxy, propoxy, butoxy, substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, substituted or unsubstituted C6-C12 aryl, and substituted or unsubstituted C4-C7 heteroaryl, wherein the substitution refers to being substituted by one or more of C1-C6 alkyl, C1-C6 alkoxy, amino, hydroxyl, halogen, nitro, and trifluoromethyl;

[0012] R2 is selected from hydroxyl or hydrogen;

[0013] X is selected from one or more of amino, optionally substituted or unsubstituted aminopiperidine, optionally substituted or unsubstituted piperazine, optionally substituted or unsubstituted homopiperazine, and optionally substituted or unsubstituted nitrogen-containing heterocycle.

[0014] Preferably, the cinnamoyl-substituted unsaturated nitrogen-containing heterocyclic oxygen-containing compound is selected from one of the following structures:

[0015]

[0016]

[0017] The application also provides a preparation method of the above-mentioned unsaturated nitrogen-containing heterocyclic compound containing a cinnamyl group or a stereoisomer, or a salt or a solvate thereof, and a synthetic route is as follows:

[0018]

[0019] In some embodiments,

[0020] R2=H, OH

[0021]

[0022] Specifically comprising the following steps:

[0023] (1) Compound V is dissolved in a solvent, dichlorosulfoxide is added thereto, and stirring reaction is carried out at room temperature, and TLC is used to track the reaction to obtain compound IV;

[0024] In the application, after the reaction is completed, the solvent is removed by rotary evaporation.

[0025] (2) Compound IV and triethylamine are dissolved in a solvent, Boc-X is added thereto, and stirring reaction is carried out at room temperature, and TLC is used to track the reaction to obtain compound III;

[0026] In the application, after the reaction is completed, the product is separated and purified, and the separation and purification specifically comprises the following steps: after the reaction is completed, heavy distilled water is added, dichloromethane is used for extraction, the organic phases are combined, then water washing, drying, and removal of the solvent, and the remaining material is purified and separated by column chromatography.

[0027] (3) Compound III is dissolved in a solvent, concentrated hydrochloric acid is added thereto, and stirring reaction is carried out at room temperature, and TLC is used to track the reaction to obtain compound II;

[0028] In the application, after the reaction is completed, the product is separated and purified, and the separation and purification specifically comprises the following steps: after the reaction is completed, heavy distilled water is added, a sodium hydroxide solution is used to adjust the pH of the water layer to 8-9 at 0℃, dichloromethane is added for extraction, the organic phases are combined, then drying, and removal of the solvent, and the remaining material is purified and separated by column chromatography.

[0029] (4) Compound VIII is dissolved in a solvent, anhydrous potassium carbonate is added, and stirring reaction is carried out with 1,3-dibromopropane and epibromohydrin at 0℃ to obtain compound VII;

[0030] Compound VIII is dissolved in a solvent, anhydrous potassium carbonate is added, and stirring reaction is carried out with 1,3-dibromopropane and epibromohydrin at 80℃ to obtain compound VI;

[0031] In the present application, after the stirring reaction is completed, the separation and purification of the product is also included, which is specifically as follows: after the stirring reaction is completed, heavy distilled water is added, dichloromethane is used for extraction, the organic phase is combined, then water washing, drying, removal of solvent, and the residue is purified and separated by column chromatography.

[0032] (5) Compound II and potassium carbonate are added into a solvent, stirring until the reactants are completely dissolved, then droping compound VII or compound VI, stirring at 70℃ for 12h to obtain the target compound.

[0033] In the present application, after the stirring reaction is completed, the separation and purification of the product is also included, which is specifically as follows: after the stirring reaction is completed, heavy distilled water is added, dichloromethane is used for extraction, the organic phase is combined, then water washing, drying, removal of solvent, and the residue is purified and separated by column chromatography.

[0034] The present application also provides a composition containing the unsaturated nitrogen-containing heterocyclic oxygen-containing compound substituted with cinnamoyl group or stereoisomer thereof, or salt or solvate thereof, and an agricultural adjuvant, fungicide, insecticide or herbicide; the dosage form of the composition is selected from the group consisting of emulsifiable concentrate (EC), powder (DP), wettable powder (WP), granule (GR), aqueous solution (AS), suspension concentrate (SC), ultra-low volume spray (ULV), soluble powder (SP), microcapsule (MC), fumigant (FU), emulsion in water (EW) or water dispersible granule (WG).

[0035] The present application also provides the use of the unsaturated nitrogen-containing heterocyclic oxygen-containing compound substituted with cinnamoyl group or stereoisomer thereof, or salt or solvate thereof, or the composition in the prevention and treatment of agricultural pests and diseases, which includes plant bacterial or fungal diseases; the plant bacterial diseases include plant leaf blight, and the plant fungal diseases include plant canker.

[0036] Preferably, the agricultural pests and diseases include plant pathogenic bacteria such as rice bacterial leaf blight pathogen, Xanthomonas oryzae pv. oryzae, Xanthomonas campestris pv. campestris, Pseudomonas syringae pv. tomato, Pseudomonas syringae pv. actinidiae, Ralstonia solanacearum, Pseudomonas syringae pv. lachrymans, Pseudomonas syringae pv. cucurbitacearum, Pseudomonas syringae, Xanthomonas sojae pv. sojae or Pseudomonas syringae pv. maculicola, and plant pathogenic fungi such as Elsinoe ampelina, Fusarium oxysporum, Phoma exigua, Colletotrichum gloeosporioides, Glomerella cingulata and Diaporthe phaseolorum.

[0037] The application also provides the method for preventing and treating agricultural pests and diseases by using the compound or the composition.

[0038] The application also provides the method for protecting plants from agricultural pests and diseases, which comprises the method step of contacting the plants with the compound or the composition.

[0039] The application also provides the method for using the compound or the composition, which comprises the method step of contacting the compound or the composition with pests or the living environment of the pests, or directly contacting the plants.

[0040] Beneficial technical effects:

[0041] The application takes cinnamic acid structure as a lead compound, and then connects nitrogen-containing heterocyclic rings such as piperazine, piperidine and homopiperazine as a connecting group, and then connects different pharmacophores, so that a series of unsaturated nitrogen-containing heterocyclic oxygen-containing compounds with cinnamoyl substitution are synthesized. The compounds have excellent inhibitory effect on plant pathogenic bacteria such as rice bacterial leaf blight, citrus canker bacteria and kiwi canker bacteria, and provide an important scientific basis for the research and development of new pesticides. DETAILED DESCRIPTION

[0042] In order to make the purpose, technical solutions and advantages of the embodiments of the application clearer, the technical solutions in the embodiments of the application will be described clearly and completely. Obviously, the described embodiments are part of the embodiments of the application, rather than all the embodiments. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative labor belong to the protection scope of the application.

[0043] As used herein, the term "alkyl" includes both branched and straight-chain saturated hydrocarbon groups having the specified number of carbon atoms. In the C1-10alkyl (or alkylene) group, for example, there are C1, C2, C3, C4, C5, C6, C7, C8, C9and C10alkyl groups. Also, for example, "C1-6alkyl" indicates an alkyl group having from 1 to 6 carbon atoms. The alkyl group can be unsubstituted or substituted, such that one or more of its hydrogen atoms are replaced by other chemical groups. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, t-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), and the like.

[0044] As used herein, the term "substituted" means the replacement of any one or more of the hydrogen atoms on the designated atom or group with the selected indicated group, provided that the designated atom's normal valence is not exceeded. If not otherwise stated, the substituent designation is to the point of attachment to the core structure. For example, it is understood that when (cycloalkyl)alkyl is a possible substituent, the point of attachment to the core structure is in the alkyl portion. As used herein, a ring double bond is a double bond formed between two adjacent ring atoms (e.g., C=C, C=N or N=N). When substitution is referred to, especially polysubstitution, it is intended to mean that multiple substituents are substituted on the indicated group at various positions, e.g., dichlorophenyl means 1,2-dichlorophenyl, 1,3-dichlorophenyl, and 1,4-dichlorophenyl.

[0045] Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds or useful synthetic intermediates. A stable compound or stable structure is meant to imply a compound that is sufficiently robust to survive isolation from a reaction mixture and subsequent formulation into an efficacious therapeutic agent. Preferably, the compound does not contain N-halogen, S(O)2H or S(O)H groups.

[0046] The term "aryl" means a monocyclic or bicyclic aromatic hydrocarbon group having from 6 to 12 carbon atoms in the ring portion, such as phenyl and naphthyl, each of which can be substituted.

[0047] The term "halogen" or "halo" means chlorine, bromine, fluorine, and iodine.

[0048] The term "haloalkyl" means a substituted alkyl group having one or more halogen substituents. For example, "haloalkyl" includes mono, di, and tri-fluoromethyl; even though the halo in haloalkyl is specifically stated to be fluorine, chlorine, bromine, iodine, it also refers to a substituted alkyl group having one or more fluorine, chlorine, bromine, iodine substituents.

[0049] The term "heteroaryl" refers to substituted and unsubstituted aromatic 5- or 6-membered monocyclic groups, 9- or 10-membered bicyclic groups, and 11- to 14-membered tricyclic groups, having at least one heteroatom (O, S, or N) in at least one ring, with the heteroatom-containing rings preferably having 1, 2, or 3 heteroatoms selected from O, S, and N. Each ring of the heteroaryl group containing a heteroatom can contain one or two oxygen or sulfur atoms and / or from 1 to 4 nitrogen atoms, provided that the total number of heteroatoms in each ring is 4 or less, and each ring has at least one carbon atom. The fused rings completing the bicyclic and tricyclic groups can contain only carbon atoms and can be saturated, partially saturated, or unsaturated. The nitrogen and sulfur atoms can be optionally oxidized and the nitrogen atoms can be optionally quaternized. The bicyclic or tricyclic heteroaryl must include at least one fully aromatic ring, and the other fused rings can be aromatic or nonaromatic. The heteroaryl group can be attached at any available nitrogen or carbon atom of any ring.

[0050] Exemplary monocyclic heteroaryls include pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, furanyl, thiophenyl, oxadiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, and the like.

[0051] Exemplary bicyclic heteroaryls include indolyl, benzothiazolyl, benzodioxolyl, benzoxazolyl, benzothiophenyl, quinolyl, tetrahydroisoquinolyl, isoquinolyl, benzimidazolyl, benzofuranyl, indolizinyl, benzofuranyl, chromonyl, coumarinyl, benzofuranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, fluoropyridyl, dihydroisoindolyl, tetrahydroquinolinyl, and the like.

[0052] If not otherwise stated, the compounds of the present application are understood to include the free form and salts thereof. The term "salt" denotes acidic and / or basic salts formed with inorganic and / or organic acids and bases. In addition, the term "salt" can include zwitterions (inner salts), such as when a compound of Formula I contains both a basic fragment, such as an amine or pyridine or imidazole ring, and an acidic fragment such as a carboxylic acid. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, such as salts of acceptable metals and amines, wherein the cation does not contribute significant toxicity or biological activity to the salt. However, other salts can be useful, such as in isolation or purification steps, and are included within the scope of the present application.

[0053] When referring to a substituent group as alkenyl, alkynyl, alkyl, halo, aryl, heteroaryl, alkoxy, cycloalkyl, hydroxy, amino, thiol, or when referring to a specific alkenyl, alkynyl, alkyl, halo, aryl, heteroaryl, alkoxy, cycloalkyl, hydroxy, amino, thiol group specifically, one to three of the above substituents are intended.

[0054] Example 1

[0055] 1) Preparation of intermediate 3-nitrocinnamoyl chloride:

[0056]

[0057] In a 50 mL round bottom flask, add compound 3-nitrocinnamic acid (0.6 g, 3.0 mmol) and 10 mL of dichlorosulfoxide, stir the reaction at room temperature, TLC track the reaction. After the reaction is completed, add 10 mL of dichloromethane in three times. Remove the solvent at 50°C by rotary evaporator to obtain intermediate 3-nitrocinnamoyl chloride;

[0058] 2) Preparation of intermediate tert-butyl 4-(3-(3-nitrophenyl)acryloyl)homopiperazine-1-carboxylate:

[0059]

[0060] Stir the mixture of intermediate 3-nitrocinnamoyl chloride (641.0 mg, 3.0 mmol), 5 mL of dichloromethane, 1-Boc-homopiperazine (610.0 mg, 3.0 mmol) and triethylamine (420 μL, 3.0 mmol) at room temperature for about 3 hours, the reaction is completed. TLC track the reaction completion. After the reaction is completed, add 100 mL of heavy water, extract with 50 mL of dichloromethane three times, combine the organic phase, dry the organic phase with anhydrous Na2SO4. Finally remove the solvent at 42°C by rotary evaporator, then purify and separate by column chromatography (eluent (V / V): PE / EA = 5:1) to obtain intermediate tert-butyl 4-(3-(3-nitrophenyl)acryloyl)homopiperazine-1-carboxylate;

[0061] 3) Preparation of intermediate 3-(3-nitrophenyl)-1-homopiperazine-1-propyl-2-enone:

[0062]

[0063] Dissolve intermediate tert-butyl 4-(3-(3-nitrophenyl)acryloyl)homopiperazine-1-carboxylate (1.1 g, 3.0 mmol) in 10 mL of ethyl acetate, then add 2 mL of concentrated hydrochloric acid in a 50 mL round bottom flask, stir the reaction for 12 h, TLC track the reaction completion. After the reaction is completed, add 100 mL of heavy water, adjust the pH of the water layer to 8-9 at 0°C with sodium hydroxide solution, then extract with 50 mL of dichloromethane three times, combine the organic phase, dry the organic phase with anhydrous Na2SO4. Finally remove the solvent at 42°C by rotary evaporator to obtain intermediate 3-(3-nitrophenyl)-1-homopiperazine-1-propyl-2-enone;

[0064] 4) Preparation of intermediate 3-phenoxybromopropanamine:

[0065]

[0066] Phenol (0.48 g, 5 mmol) and potassium carbonate (2.79 g, 20 mmol) were dissolved in 5 mL DMF and stirred in an ice bath, then 1,3-dibromopropane (4.38 mL, 40 mmol) was slowly added dropwise, and stirring was continued for about 6 h, the reaction was completed, TLC tracking reaction completion. After the reaction was completed, 100 mL of heavy water was added, extracted with 50 mL of dichloromethane three times, the organic phase was combined and dried over anhydrous Na2SO4. Finally, the solvent was removed by rotary evaporator at 42°C, and then purified by column chromatography (eluent (V / V): PE / EA = 5:1) to obtain the intermediate 3-phenoxybromopropane ethylamine;

[0067] 5) Preparation of 3-(3-nitrophenyl)-1-(4-(3-phenoxypropyl)homopiperazin-1- propyl)2-alken-1-one:

[0068]

[0069] A mixture of intermediate 3-(3-nitrophenyl)-1-homopiperazine-1-propyl-2-alkenone (170.0 mg, 0.6 mmol), acetonitrile 5 mL, potassium carbonate (140.0 mg, 10.0 mmol) and 3-phenoxybromopropane ethylamine (170 mg, 0.6 mmol) was stirred at 70°C for about 12 hours, the reaction was completed. The mixture was added to 50 mL of heavy water, extracted with dichloromethane three times, dried over anhydrous Na2SO4 and concentrated in vacuum, then purified by thin layer chromatography (eluent (V / V): DCM / MeOH = 15:1) to obtain orange oil, yield 38.1%. Its NMR data are: 1 H NMR (500 MHz, CDC13) δ 8.36 (dd, J = 10.7, 1.8 Hz, 1H, phenyl-H), 8.24-8.12 (m, 1H, phenyl-H), 7.73 (ddd, J = 25.6, 16.2, 9.1 Hz, 2H, phenyl-H), 7.54 (td, J = 7.9, 4.8 Hz, 1H, phenyl-CH=), 7.31-7.17 (m, 1H, 2H, phenyl-CH= CH -+phenyl-H), 7.05-6.77 (m, 4H, phenyl-H), 3.99 (td, J = 6.2, 2.6 Hz, 2H, -O- CH 2-), 3.81-3.68 (m, 4H, Homopiperazinyl-H+-O-CH2-CH2- CH2-), 2.86 - 2.62 (m, 6H, Homopiperazinyl-H), 1.94 (td, J = 14.5, 6.7 Hz, 4H, Homopiperazinyl-H+-O-CH2- CH 2-); 13 C NMR (126 MHz, CDC13) δ 165.57, 158.97, 148.69, 140.11, 139.96, 137.15, 134.26, 129.97, 129.53, 124.02, 121.66, 120.73, 114.52, 65.83, 56.70, 55.48, 54.77, 47.24, 45.15, 29.80, 27.22. HRMS (ESI) [M+H] + calcd for C 23 H 27 N3O4: 410.2074, found: 410.2065.

[0070] The structures of the synthesized cinnamoyl group-substituted unsaturated nitrogen-containing heterocyclic oxygen-containing compounds and the nuclear magnetic resonance hydrogen spectrum and carbon spectrum data are shown in Table 1, and the physicochemical properties are shown in Table 2.

[0071] Table 1 Nuclear magnetic resonance hydrogen spectrum, carbon spectrum and high resolution mass spectrum data of the compounds

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089] Table 2 Physicochemical properties of the target compounds

[0090]

[0091]

[0092] Pharmacological Example 1

[0093] EC 50 Median effective concentration (MEC) is an important indicator for evaluating the sensitivity of plant pathogens to compounds, and it is also a crucial parameter for setting the compound concentration when studying the mechanism of action of target compounds. In concentration gradient experiments, five appropriate concentrations were set using the two-fold dilution method. Finally, the inhibition rate of the agent against the plant pathogen and the agent concentration were converted into logarithmic values, and the toxicity curve was obtained through regression analysis using SPSS software to calculate the EC50. 50 .

[0094] The effective medium concentration (EC) of the target compound against plant pathogens was determined using a turbidimetric method. 50 The experimental subjects were *Xoo*, *Xac*, and *Xoc*, the pathogens of rice bacterial leaf blight. DMSO was dissolved in the culture medium as a blank control. *Xoo* (the pathogen of rice bacterial leaf blight was on M210 solid medium) was placed in NB medium and cultured in a constant temperature shaker at 28℃ and 180 rpm until the logarithmic growth phase. *Xoc* (the pathogen of rice bacterial leaf blight was on M210 solid medium) was placed in NB medium. *Xoc* (the pathogen of rice bacterial leaf blight was on M210 solid medium) was placed in NB medium and cultured in a constant temperature shaker at 28℃ and 180 rpm until the logarithmic growth phase. Prepare different concentrations (e.g., 100, 50, 25, 12.5, 6.25 μg / mL) of the drug (compound) in 5 mL of NB liquid culture medium containing the pathogen and add them to test tubes. Add 40 μL of NB liquid culture medium containing plant pathogen bacteria to each tube. Incubate the tubes at 28-30℃ and 180 rpm in a constant temperature shaker. Culture the pathogens for rice bacterial blight, citrus canker, and rice bacterial leaf streak for 48 h, respectively. Measure the OD of each concentration of bacterial culture using a microplate reader. 595The OD value of the sterile NB liquid culture medium containing the corresponding concentration of the toxin was additionally measured 595 .

[0095] Corrected OD value = OD value of the culture medium containing bacteria - OD value of the sterile culture medium

[0096] Inhibition rate % = [(corrected OD value of the control culture medium bacterial liquid - corrected OD value of the culture medium containing toxin) / corrected OD value of the control culture medium bacterial liquid] x 100

[0097] The embodiments of the present application assist in illustrating the technical solutions of the present application, but the contents of the embodiments are not limited thereto. The experimental results of the target compounds are shown in Table 3.

[0098] Table 3 EC of oxygen-containing compounds containing cinnamoyl-substituted unsaturated nitrogen-containing heterocycles on plant pathogenic bacteria 50

[0099]

[0100]

[0101]

[0102] As can be seen from Table 3, in the in vitro test, the target compounds showed good bacteriostatic activity on plant pathogenic bacteria (such as Xanthomonas oryzae pv. oryzae, Xac and Xanthomonas oryzae pv. oryzicola, Xoc). Compounds 8, 9, 32 and 33 showed excellent inhibitory activity on Xanthomonas oryzae pv. oryzae (Xoo), with an EC 50 of 0.034-0.53 μg / m L; compounds 8, 9, 32, 33, 40 and 41 showed excellent inhibitory activity on Xanthomonas axonopodis pv. citri (Xac), with an EC 50 of 0.034-0.78 μg / m L; compounds 24, 32, 33, 40, 56 showed excellent inhibitory activity on Xanthomonas oryzae pv. oryzicola (Xoc), with an EC 50 of 0.026-0.95 μg / m L. They can be used for preparing pesticides against plant pathogenic bacteria.

[0103] Pharmacological Example 2 Inhibitory activity of the compounds on plant pathogenic fungi (50 μg / mL)

[0104] The mycelium growth rate method, also known as the toxic medium method, is one of the conventional methods for determining the toxicity of fungicides. The main principle is to mix the test agent with the culture medium, and to measure the toxicity of the agent by the growth rate of the colonies on the toxic culture medium. In this example, the test objects are Botryosphaeria dothidea, Fusarium SP, and Physoderma sorokinianum, and DMSO is used as a blank control. The specific operation is as follows: 1) according to the test concentration, weigh an appropriate amount of drug, dissolve it with DMSO (the amount should not exceed 1% of the final toxic culture medium), then add 0.1% Tween 20 solution to make up to 10 mL, pour into 90 mL of melted PDA culture medium, mix well, and pour into 9 culture dishes for standby; 2) sterilize the puncher (inner diameter of 5 mm) by burning, cool it, and then punch the mycelium at the edge of the pre-activated strain, and use the inoculation needle to paste the mycelium surface to the center of the toxic culture medium. After the treatment is completed, place them in a 25°C incubator; 3) when the colony diameter of the control group grows to 5.5-6.6 cm, measure the colony diameters of the control group and each drug treatment group using the cross method; 4) calculate the inhibition rate (%) using the following formula: inhibition rate (%) = (C-T) / (C-0.5) x 100; where C is the colony diameter of the control group, T is the colony diameter of the drug treatment group, and 0.5 is the diameter of the inoculated fungus cake.

[0105] According to the above method, the experimental results of the target compounds in Table 1 are shown in Table 4.

[0106] Table 4 Inhibition activity of compounds on plant pathogenic fungi (50 μg / mL)

[0107]

[0108] As can be seen from Table 4, in the in vitro test, the target compounds showed good inhibition activity on plant pathogenic fungi (such as Botryosphaeria dothidea, Fusarium SP, and Physoderma sorokinianum). Compounds 4 and 7 showed good inhibition activity on Physoderma sorokinianum, with inhibition rates of 60.38% and 62.26% at 50 μg / mL, respectively. Compound 4 showed comparable inhibition activity on Botryosphaeria dothidea to the control drug, amylcin, with inhibition rates of 43.33% and 42.73% at 50 μg / mL, respectively. Compound 7 showed higher inhibition activity on Fusarium SP than the control drug, amylcin, with inhibition rates of 30.61% and 18.87%, respectively. It is shown that compounds 4 and 7 have certain inhibition effect on plant fungal diseases.

[0109] The above only describes the preferred embodiments of the present application, and it should be noted that those skilled in the art can make several improvements and refinements without departing from the principles of the present application, and these improvements and refinements should also be considered within the protection scope of the present application.

Claims

1. An unsaturated cinnamoyl-substituted nitrogen-containing heterocyclic oxygen-containing compound having the structure of Formula I: or a salt thereof, wherein: R1 is selected from substituted or unsubstituted phenyl, wherein substitution means substituted with one or more of C1-C6 alkyl, C1-C6 alkoxy, halogen, nitro, trifluoromethyl; R2 is selected from hydroxyl or hydrogen; R3 is selected from phenyl, substituted or unsubstituted coumarinyl, wherein substitution means substituted with C1-C6 alkyl or halogen; X is selected from piperazine or homopiperazine. wherein The unsaturated cinnamoyl-substituted nitrogen-containing heterocyclic oxygen-containing compound is selected from one of the following structures: The compound II and potassium carbonate are added to a solvent, stirred until the reactants are completely dissolved, then compound VII or compound VI is added dropwise, stirred at 70°C for 12h to obtain the target compound; The preparation method of the compound II comprises the following steps: (1) dissolving compound V in a solvent, then adding dichlorosulfoxide thereto, stirring at room temperature, tracking the reaction by TLC to obtain compound IV; 2. An unsaturated nitrogen-containing heterocyclic oxygen-containing compound or a salt thereof containing a cinnamoyl group, characterized by (2) dissolving compound IV and triethylamine in a solvent, then adding Boc-X thereto, stirring at room temperature, tracking the reaction by TLC to obtain compound III; 3. A process for the production of the cinnamoyl group-containing unsaturated nitrogen-containing heterocycle-containing oxygen compound or salt thereof according to Claim 1 or 2, characterized by, (3) dissolving compound III in a solvent, then adding concentrated hydrochloric acid thereto, stirring at room temperature, tracking the reaction by TLC to obtain compound II; The structural formula of the compound II is The structural formula of the compound VII is The structural formula of the compound VI is 4. The production method according to claim 3, characterized by, The preparation method of the compound VII is: dissolving compound VIII in a solvent, adding anhydrous potassium carbonate, stirring with 1,3-dibromopropane at 0°C to obtain compound VII; The preparation method of the compound VI is: dissolving compound VIII in a solvent, adding anhydrous potassium carbonate, stirring with epoxy bromopropane at 80°C to obtain compound VI; The structural formula of compound V is The structural formula of compound IV is The compound VIII has the structural formula of R3-OH. The structural formula of compound III is The unsaturated cinnamoyl-substituted nitrogen-containing heterocyclic oxygen-containing compound or salt thereof of claim 1 or 2, and an agricultural adjuvant, fungicide, insecticide or herbicide; the dosage form of the composition is selected from the group consisting of emulsifiable concentrate, powder, granule, aqueous agent, suspension concentrate, ultra-low volume spray, microcapsule, smoke agent, and water emulsion. The structural formula of compound II is The agricultural pests are Xanthomonas oryzae pv. oryzae, Botryosphaeria dothidea, Fusarium oxysporum, Physoderma sorokinianum, and Xanthomonas axonopodis pv. citri. The method comprises the step of allowing the harmful organisms or their living environment to be affected by the unsaturated cinnamoyl-substituted nitrogen-containing heterocyclic oxygen-containing compound or salt thereof of claim 1 or 2, or the composition of claim 5; and the agricultural pests are Xanthomonas oryzae pv. oryzae, Botryosphaeria dothidea, Fusarium oxysporum, Physoderma sorokinianum, and Xanthomonas axonopodis pv. citri. The method comprises the step of allowing the harmful organisms or their living environment to be affected by the unsaturated cinnamoyl-substituted nitrogen-containing heterocyclic oxygen-containing compound or salt thereof of claim 1 or 2, or the composition of claim 5; and the agricultural pests are Xanthomonas oryzae pv. oryzae, Botryosphaeria dothidea, Fusarium oxysporum, Physoderma sorokinianum, and Xanthomonas axonopodis pv. citri.

5. A composition characterized in that, ​ 6. Use of the cinnamoyl group-containing unsaturated nitrogen-containing heterocycle- containing oxygen compound or salt thereof according to claim 1 or 2, or the composition according to claim 5, for controlling agricultural pests and diseases, characterized in that, ​ 7. The cinnamoyl group-containing unsaturated nitrogen-containing heterocycle- containing oxygen compound or salt thereof according to Claim 1 or 2, or the method for controlling agricultural pests and diseases by using the composition according to Claim 5, characterized in that, ​ 8. A method of protecting a plant from an agricultural pest or disease, characterized in that, ​

Images