A polysubstituted nitrogen-containing heterocyclic derivative, a preparation method and application thereof

Abstract

The application provides a polysubstituted nitrogen-containing heterocyclic derivative and a preparation method and application thereof. The polysubstituted nitrogen-containing heterocyclic derivative is obtained by designing a novel structure based on a natural nitrogen-containing heterocyclic molecule, and the polysubstituted nitrogen-containing heterocyclic derivative has obvious bactericidal activity, and some compounds have significant bactericidal activity on soft rot bacteria, escherichia coli, pseudomonas solanacearum and other diseases, and can be widely applied to the comprehensive prevention and control of bacterial diseases in agriculture and forestry. The bactericide obtained by mixing the polysubstituted nitrogen-containing heterocyclic derivative of the application with different additives also has good bactericidal effect on escherichia coli, soft rot bacteria, pseudomonas solanacearum and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of biopharmaceuticals, and in particular to a polysubstituted nitrogen-containing heterocyclic derivative and a preparation method and application thereof. Background Art

[0002] Bacterial plant diseases are diseases caused by a series of physiological changes in plant cells and tissues caused by bacterial pathogens. These diseases include soft rot, black rot, canker, bacterial wilt, leaf blight, and leaf spot. In recent years, bacterial diseases have become the second most common disease in Chinese agriculture, second only to fungal diseases. They occur in vegetables, fruit trees, rice, and a variety of other crops, and multiple bacterial diseases can occur within the same plant species. Bacterial diseases often spread in diverse ways, with explosive, epidemic, and devastating effects, making their prevention and control increasingly difficult. For example, bacterial wilt is a widespread soil-borne bacterial disease worldwide. Soil acidification and compaction can lead to widespread wilt outbreaks, and treatment is extremely difficult in the middle and late stages of the disease. Bacterial wilt can infect up to 200 crop species across 50 families, causing widespread death and significant yield reductions in crops such as tomatoes, peppers, ginger, and tobacco, and even destroying orchards. Each year, bacterial wilt causes significant losses in potato production.

[0003] Common bacterial crop diseases include soft rot, canker, bacterial wilt, rice bacterial leaf blight, rice bacterial leaf streak, soybean bacterial blight, and black rot. Bacterial wilt, a devastating vascular disease of Solanaceae plants like peppers, tomatoes, and tobacco, can cause yield losses of 20-50%. Currently, the number and variety of pesticides available for bacterial disease control in my country is very limited, and most are copper preparations and antibiotics. However, long-term use has led to increasing bacterial resistance, and the effectiveness of existing pesticides has been declining.

[0004] Since there are currently few and problematic agents for preventing and controlling bacterial diseases, it is necessary to improve them. Summary of the Invention

[0005] In view of this, the present invention proposes a polysubstituted nitrogen-containing heterocyclic derivative and a preparation method and application thereof to solve the technical defects existing in the prior art.

[0006] In a first aspect, the present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the structural formula of the polysubstituted nitrogen-containing heterocyclic derivative is as follows:

[0007]

[0008] Among them, the R 1One selected from the following groups: H, CH3, CH3CH2, CH3O, OH, F, Cl, Br, CN, CF3, CF3O, NO2, 1-methyl-piperazin-4-yl, morpholine-4-yl, piperazin-1-yl;

[0009] The R 2 One selected from the following groups: H, CH3, CH3CH2, CH3O, OH, F, Cl, Br, CN, CF3, CF3O, NO2;

[0010] The R 3 One selected from the following groups: H, CH3, CH3CH2, CH3O, OH, F, Cl, Br, CN, CF3, CF3O, NO2;

[0011] The R 4 One selected from the following groups: H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, tBu, CH(CH2)2, CH2C≡CH, CH2CH=CH2, CH3O, CH2CH2SO2CH3, CH2CH2SCH3, CH2CN, CH2CF3, CH2CONH2, C(CH2)2CN, CH2Ph, CH2CH2Ph, CH2-4-MeO-Ph, CH2CH2-4-MeO-Ph, CH2-3-MeO-Ph, CH2CH2-3-MeO-Ph, CH2-4-F-Ph, CH2C H2-4-F-Ph, CH2-4-CF3-Ph, CH2CH2-4-CF3-Ph, CH(CH3)Ph, CH2-4-OH-3-MeO-Ph, Ph, 4-MeO-Ph, 4-CF3-Ph, 3-MeO-Ph, 3-CF3-Ph, 2-MeO- Ph, 2-CF3-Ph, 4-Me-Ph, 2-Me-Ph, 3-Me-Ph, 4-Cl-Ph, 3-Cl-Ph, 2-Cl-Ph, 4-F-Ph, 3-F-Ph, 2-F-Ph, 4-Br-Ph, 2-F-4-Cl-Ph, 2,4-Cl2-Ph;

[0012] The R 5One selected from the following groups: CO2Et, CO2Me, COOH, CONHCH2CH3, CONHCH(CH2)2, CONHC(CH2)2CN, CONHCH2CH2SO2CH3, CONHCH2CH2SCH3, CONHNHCOPh, CONHNHCO-4-Py, CONHCH2Ph, CONHCH2CH2Ph, CONHCH2-4-MeO-Ph, CONHCH2CH2-4-MeO-Ph, CONHCH2-4-OH-3-MeO-Ph, CONHCH2CH2-4-F-Ph, SPh, SO2Ph, SO2-4-Me-Ph, SO2-4-Cl-Ph, SO2-2-Me-Ph, SO2-2-Cl-Ph, SO2-3-Me-Ph, SO2-3-Cl-Ph,

[0013] The X is one of CH or N.

[0014] In a second aspect, the present invention further provides a method for preparing the polysubstituted nitrogen-containing heterocyclic derivative, comprising the following steps:

[0015] Substituted aryl methyl ketone is used as a raw material and reacted with diethyl carbonate or dimethyl carbonate under alkaline conditions to prepare substituted aryl formyl acetate;

[0016] Condensing the substituted aryl formyl acetate with N,N-dimethylformamide dimethyl acetal to obtain a first intermediate;

[0017] reacting the first intermediate with a substituted amine under alkaline conditions to obtain a first polysubstituted nitrogen-containing heterocyclic derivative;

[0018] hydrolyzing the first polysubstituted nitrogen-containing heterocyclic derivative under acidic or alkaline conditions to obtain a second polysubstituted nitrogen-containing heterocyclic derivative;

[0019] The second polysubstituted nitrogen-containing heterocyclic derivative and the substituted amine are subjected to a condensation reaction in the presence of a condensing agent to obtain a third polysubstituted nitrogen-containing heterocyclic derivative;

[0020] Alternatively, a substituted aryl methyl ketone is used as a raw material and a brominated intermediate is obtained by bromination reaction;

[0021] The brominated intermediate is reacted with sodium substituted benzenesulfinate or sodium substituted benzenethiophenol to obtain a second intermediate or a third intermediate;

[0022] condensing the second intermediate or the third intermediate with N,N-dimethylformamide dimethyl acetal to obtain a fourth intermediate or a fifth intermediate;

[0023] The fourth intermediate or the fifth intermediate undergoes a substitution cyclization reaction with a substituted amine under the action of a base to obtain a fourth polysubstituted nitrogen-containing heterocyclic derivative or a fifth polysubstituted nitrogen-containing heterocyclic derivative;

[0024] Wherein, the structural formula of the substituted aryl methyl ketone is:

[0025] The structural formula of the substituted aryl formyl acetate is:

[0026] The structural formula of the first intermediate is:

[0027] The structural formula of the first polysubstituted nitrogen-containing heterocyclic derivative is:

[0028] The structural formula of the second multi-substituted nitrogen-containing heterocyclic derivative is:

[0029] The structural formula of the third polysubstituted nitrogen-containing heterocyclic derivative is:

[0030] The structural formula of the brominated intermediate is:

[0031] The structural formula of the second intermediate is:

[0032] The structural formula of the third intermediate is:

[0033] The structural formula of the fourth intermediate is:

[0034] The structural formula of the fifth intermediate is:

[0035] The structural formula of the fourth polysubstituted nitrogen-containing heterocyclic derivative is: The structural formula of the fifth polysubstituted nitrogen-containing heterocyclic derivative is:

[0036] Wherein, the R group is one of Me and Et.

[0037] In a third aspect, the present invention further provides a fungicide comprising the polysubstituted nitrogen-containing heterocyclic derivative.

[0038] In a fourth aspect, the present invention further provides a use of the polysubstituted nitrogen-containing heterocyclic derivative or the fungicide in preventing and controlling plant bacterial diseases.

[0039] A polysubstituted nitrogen-containing heterocyclic derivative of the present invention has the following beneficial effects compared with the existing ones:

[0040] The polysubstituted nitrogen-containing heterocyclic derivatives of the present invention have obvious fungicidal activity. Some compounds have significant fungicidal activity against diseases such as soft rot fungi, Escherichia coli, and Ralstonia solanacearum, and can be widely used in the comprehensive prevention and control of agricultural and forestry bacterial diseases. The fungicides obtained by mixing the polysubstituted nitrogen-containing heterocyclic derivatives of the present invention with different adjuvants also have good fungicidal effects against Escherichia coli, soft rot fungi, Ralstonia solanacearum, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] To more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. Those skilled in the art can also derive other drawings based on these drawings without inventive effort.

[0042] Figure 1 The results of the preliminary screening of the fungicidal activity of different compounds against Ralstonia solanacearum in Example 1 of the present invention are as follows;

[0043] Figure 2 The results of the bactericidal activity screening of compounds at different concentrations against Escherichia coli are shown;

[0044] Figure 3 The results of fungicidal activity screening of compounds at different concentrations against soft rot fungi are shown;

[0045] Figure 4 These are the results of fungicidal activity screening of compounds at different concentrations against Ralstonia solanacearum. DETAILED DESCRIPTION

[0046] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0047] The present application provides a polysubstituted nitrogen-containing heterocyclic derivative, the structural formula of which is shown below:

[0048]

[0049] Among them, R 1is selected from one of the following groups: H, CH3, CH3CH2, CH3O, OH, F, Cl, Br, CN, CF3, CF3O, NO2, 1-methyl-piperazin-4-yl (i.e., 1-methyl-piperazin-4-yl), morpholine-4-yl (morpholine-4-yl), piperazin-1-yl (i.e., (piperazin-1-yl); R 2 One selected from the following groups: H, CH3, CH3CH2, CH3O, OH, F, Cl, Br, CN, CF3, CF3O, NO2;

[0050] R 3 One selected from the following groups: H, CH3, CH3CH2, CH3O, OH, F, Cl, Br, CN, CF3, CF3O, NO2;

[0051] R 4 One selected from the following groups: H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, tBu, CH(CH2)2, CH2C≡CH, CH2CH=CH2, CH3O, CH2CH2SO2CH3, CH2CH2SCH3, CH2CN, CH2CF3, CH2CONH2, C(CH2)2CN, CH2Ph, CH2CH2Ph, CH2-4-MeO-Ph, CH2CH2-4-MeO-Ph, CH2-3-MeO-Ph, CH2CH2-3-MeO-Ph, CH2-4-F-Ph, CH2C H2-4-F-Ph, CH2-4-CF3-Ph, CH2CH2-4-CF3-Ph, CH(CH3)Ph, CH2-4-OH-3-MeO-Ph, Ph, 4-MeO-Ph, 4-CF3-Ph, 3-MeO-Ph, 3-CF3-Ph, 2-MeO- Ph, 2-CF3-Ph, 4-Me-Ph, 2-Me-Ph, 3-Me-Ph, 4-Cl-Ph, 3-Cl-Ph, 2-Cl-Ph, 4-F-Ph, 3-F-Ph, 2-F-Ph, 4-Br-Ph, 2-F-4-Cl-Ph, 2,4-Cl2-Ph;

[0052] R 5One selected from the following groups: CO2Et, CO2Me, COOH, CONHCH2CH3, CONHCH(CH2)2, CONHC(CH2)2CN, CONHCH2CH2SO2CH3, CONHCH2CH2SCH3, CONHNHCOPh, CONHNHCO-4-Py, CONHCH2Ph, CONHCH2CH2Ph, CONHCH2-4-MeO-Ph, CONHCH2CH2-4-MeO-Ph, CONHCH2-4-OH-3-MeO-Ph, CONHCH2CH2-4-F-Ph, SPh, SO2Ph, SO2-4-Me-Ph, SO2-4-Cl-Ph, SO2-2-Me-Ph, SO2-2-Cl-Ph, SO2-3-Me-Ph, SO2-3-Cl-Ph,

[0053] X is one of CH or N.

[0054] In the above embodiments, tBu represents a tert-butyl group, the group represented by Ph is a phenyl group, Me represents a methyl group, Et represents an ethyl group, and Py represents a pyridyl group.

[0055] Nitrogen-containing heterocyclic compounds are common molecular skeletons widely present in natural products and synthetic drugs. They usually have important physiological and biochemical functions and have very important applications in pharmaceutical chemistry, agricultural and food chemistry, biochemistry, and polymer materials. This application uses natural nitrogen-containing heterocyclic molecules as a guide to carry out the design of novel structures to obtain polysubstituted nitrogen-containing heterocyclic derivatives. Polysubstituted nitrogen-containing heterocyclic derivatives have significant fungicidal activity. Some compounds have significant fungicidal activity against diseases such as soft rot fungi, Escherichia coli, and Ralstonia solanacearum, and can be widely used in the comprehensive prevention and control of agricultural and forestry bacterial diseases.

[0056] The polysubstituted nitrogen-containing heterocyclic derivatives of the present invention also include stereoisomers, tautomers, solvates or pesticide-acceptable salts of the compounds represented by the formula.

[0057] In some embodiments, the polysubstituted nitrogen-containing heterocyclic derivative is selected from any one of the following compounds:

[0058]

[0059]

[0060] Based on the same inventive concept, the present invention also provides a method for preparing the above-mentioned polysubstituted nitrogen-containing heterocyclic derivative, comprising the following steps:

[0061] Substituted aryl methyl ketone is used as a raw material and reacted with diethyl carbonate or dimethyl carbonate under alkaline conditions to prepare substituted aryl formyl acetate;

[0062] condensing a substituted aryl formyl acetate with N,N-dimethylformamide dimethyl acetal to obtain a first intermediate;

[0063] reacting the first intermediate with a substituted amine under alkaline conditions to obtain a first polysubstituted nitrogen-containing heterocyclic derivative;

[0064] hydrolyzing the first polysubstituted nitrogen-containing heterocyclic derivative under acidic or alkaline conditions to obtain a second polysubstituted nitrogen-containing heterocyclic derivative;

[0065] The second polysubstituted nitrogen-containing heterocyclic derivative and the substituted amine are subjected to a condensation reaction in the presence of a condensing agent to obtain a third polysubstituted nitrogen-containing heterocyclic derivative;

[0066] Alternatively, a substituted aryl methyl ketone is used as a raw material and a brominated intermediate is obtained by bromination reaction;

[0067] The brominated intermediate is reacted with sodium substituted benzenesulfinate or sodium substituted benzenethiophenol to obtain a second intermediate or a third intermediate;

[0068] condensing the second intermediate or the third intermediate with N,N-dimethylformamide dimethyl acetal to obtain a fourth intermediate or a fifth intermediate;

[0069] The fourth intermediate or the fifth intermediate undergoes a substitution cyclization reaction with a substituted amine under the action of a base to obtain a fourth polysubstituted nitrogen-containing heterocyclic derivative or a fifth polysubstituted nitrogen-containing heterocyclic derivative;

[0070] Among them, the structural formula of the substituted aryl methyl ketone is:

[0071] The structural formula of substituted aryl formyl acetate is:

[0072] The structural formula of the first intermediate is:

[0073] The structural formula of the first polysubstituted nitrogen-containing heterocyclic derivative is:

[0074] The structural formula of the second polysubstituted nitrogen-containing heterocyclic derivative is:

[0075] The structural formula of the third polysubstituted nitrogen-containing heterocyclic derivative is:

[0076] The structural formula of the brominated intermediate is:

[0077] The structural formula of the second intermediate is:

[0078] The structural formula of the third intermediate is:

[0079] The structural formula of the fourth intermediate is:

[0080] The structural formula of the fifth intermediate is:

[0081] The structural formula of the fourth polysubstituted nitrogen-containing heterocyclic derivative is:

[0082] The structural formula of the fifth polysubstituted nitrogen-containing heterocyclic derivative is:

[0083] Wherein, the R group is one of Me and Et.

[0084] Specifically, the reaction formula occurring during the preparation of the above-mentioned polysubstituted nitrogen-containing heterocyclic derivatives is:

[0085]

[0086] Specifically, the substituted aryl methyl ketone 1 is reacted to obtain a substituted aryl formyl acetate 2; the substituted aryl formyl acetate 2 is condensed with N,N-dimethylformamide dimethyl acetal to obtain a first intermediate 3; the first intermediate 3 is reacted with a substituted amine under alkaline conditions to obtain a first polysubstituted nitrogen-containing heterocyclic derivative 4; the first polysubstituted nitrogen-containing heterocyclic derivative 4 is hydrolyzed under acidic or alkaline conditions to obtain a second polysubstituted nitrogen-containing heterocyclic derivative 5; the second polysubstituted nitrogen-containing heterocyclic derivative 5 is condensed with a substituted amine under the action of a condensing agent to obtain a third polysubstituted nitrogen-containing heterocyclic derivative 6.

[0087] Alternatively, the substituted aryl methyl ketone 1 undergoes bromination reaction to obtain a brominated intermediate 7; the brominated intermediate 7 undergoes a substitution reaction with a substituted sodium benzenesulfinate or a substituted sodium benzenethiophenol to obtain a second intermediate 8 or a third intermediate 11; the second intermediate 8 or the third intermediate 11 is condensed with N,N-dimethylformamide dimethyl acetal to obtain a fourth intermediate 9 or a fifth intermediate 12; the fourth intermediate 9 or the fifth intermediate 12 undergoes a substitution cyclization reaction with a substituted amine under the action of an alkaline agent to obtain a fourth polysubstituted nitrogen-containing heterocyclic derivative 10 or a fifth polysubstituted nitrogen-containing heterocyclic derivative 13.

[0088] In some embodiments, in the step of preparing a substituted aryl methyl ketone by reacting it with diethyl carbonate or dimethyl carbonate under alkaline conditions to obtain a substituted aryl formyl acetate, the reaction temperature is 0-50° C., the reaction solvent is one of acetonitrile, 1,2-dichloroethane, and tetrahydrofuran, and the base is one of sodium hydroxide, potassium hydroxide, or sodium hydride.

[0089] In some embodiments, in the step of condensing a substituted aromatic acetic acid ester with N,N-dimethylformamide dimethyl acetal to obtain a first intermediate, the reaction temperature is 20-100° C., the reaction solvent is one of acetonitrile, 1,2-dichloroethane, toluene, xylene, and chlorobenzene, and the catalyst is acetic acid.

[0090] In some embodiments, in the step of reacting the first intermediate with a substituted amine under alkaline conditions to obtain a first polysubstituted nitrogen-containing heterocyclic derivative, the reaction temperature is 25-60° C., the reaction solvent is one of acetonitrile, 1,2-dichloroethane, toluene, xylene, chlorobenzene, and dioxane, and the base is one of sodium hydroxide, potassium hydroxide, or sodium hydride.

[0091] In some embodiments, in the step of hydrolyzing the first polysubstituted nitrogen-containing heterocyclic derivative under acidic or alkaline conditions to obtain the second polysubstituted nitrogen-containing heterocyclic derivative, the reaction solvent is one of methanol, ethanol, isopropanol, and tetrahydrofuran; the base is one of sodium hydroxide, potassium hydroxide, and potassium carbonate; and the acid is hydrochloric acid.

[0092] In some embodiments, in the step of condensing the second polysubstituted nitrogen-containing heterocyclic derivative with the substituted amine in the presence of a condensing agent to obtain a third polysubstituted nitrogen-containing heterocyclic derivative, the reaction temperature is 20 to 50° C., the reaction solvent is any one of acetonitrile, 1,2-dichloroethane, N,N-dimethylformamide, N,N-dimethylacetamide, tetrahydrofuran, dichloromethane, and ethyl acetate, and the condensing agent is TBTU (O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate), HOBt / EDCI (1-hydroxybenzotriazole / 1 -(3-dimethylaminopropyl)-3-ethylcarbodiimide, both of which are used as condensing agents at the same time), HATU (2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate), HDTU, DCC / DMAP (dicyclohexylcarbodiimide / 4-dimethylaminopyridine, both of which are used as condensing agents at the same time), DIC / DMAP (diisopropylcarbodiimide / 4-dimethylaminopyridine, both of which are used as condensing agents at the same time), EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide).

[0093] In some embodiments, in the step of obtaining a brominated intermediate by bromination reaction using a substituted aromatic methyl ketone as a raw material, the reaction temperature is 10-60° C., the reaction solvent is one of acetonitrile, tetrahydrofuran, carbon tetrachloride, 1,2-dichloroethane, and dichloromethane, and the bromination reagent is one of N-bromosuccinimide, copper bromide, sodium bromide, bromine, and dibromohydantoin.

[0094] In some embodiments, in the step of reacting the brominated intermediate with substituted sodium benzenesulfinate or substituted sodium benzenethiophenol to obtain a second intermediate or a third intermediate, the reaction temperature is 40-90° C., and the reaction solvent is one of acetonitrile, ethanol, methanol, acetone, ethyl acetate, 1,2-dichloroethane, tetrahydrofuran, N,N-dimethylformamide, and N,N-dimethylacetamide.

[0095] In some embodiments, in the step of condensing the second intermediate or the third intermediate with N,N-dimethylformamide dimethyl acetal to obtain the fourth intermediate or the fifth intermediate, the reaction temperature is 20-100° C., the reaction solvent is one of acetonitrile, 1,2-dichloroethane, toluene, xylene, and chlorobenzene, and the catalyst is acetic acid.

[0096] In some embodiments, in the step of reacting the fourth intermediate or the fifth intermediate with a substituted amine to undergo a substitution cyclization reaction to obtain a fourth polysubstituted nitrogen-containing heterocyclic derivative or a fifth polysubstituted nitrogen-containing heterocyclic derivative, the reaction temperature is 50-150° C., the reaction solvent is one of dioxane, toluene, xylene, chlorobenzene, dimethyl sulfoxide, N,N-dimethylformamide, and N,N-dimethylacetamide, and the base is one of potassium carbonate, cesium carbonate, and sodium carbonate.

[0097] Based on the same inventive concept, an embodiment of the present application further provides a fungicide comprising the above-mentioned multi-substituted nitrogen-containing heterocyclic derivative.

[0098] In some embodiments, an auxiliary agent is further included, and the mass fraction of the polysubstituted nitrogen-containing heterocyclic derivative in the fungicide is 0.5 to 99.99%, preferably 5 to 45%.

[0099] In some embodiments, the auxiliary agent includes one or more of isopropyl alcohol, glycerol, turpentine, propanol, xylene, chlorobenzene, 1,2-dichloroethane, 1,2-dibromoethane, toluene, methanol, ethanol, N,N-dimethylformamide, ethyl acetate, acetone, butanone, cyclohexanone, dimethyl sulfoxide, and paraffin;

[0100] Or, the additive includes one or more of kaolin, bentonite, clay, diatomaceous earth, montmorillonite, activated clay, dolomite, quartz, calcium carbonate, oxide film, talc, and attapulgite;

[0101] Or, the adjuvant includes one or more of alkyl sulfonates, alkyl sulfonic acid esters, alkyl aryl sulfonates, sorbitol polyoxyethylene esters, polyoxyethylene-fatty alcohol ethers, polyoxyethylene-fatty acid esters, aralkyl polyethylene glycol ethers, fluoroalkyl sulfonic acid esters, alkyl sulfates, lignin sulfonates, polyethylene glycol, and rhamnolipids;

[0102] Or, the auxiliary agent includes one or more of polyvinyl alcohol, carboxymethyl cellulose, and gum arabic;

[0103] Alternatively, the auxiliary agent includes one or more of inorganic dyes, organic dyes and trace nutrients.

[0104] In some embodiments, the fungicide can be prepared as any one of water-dispersible granules, aqueous emulsions, suspensions, wettable powders, or nano-controlled release agents.

[0105] Based on the same inventive concept, the embodiments of the present application also provide a use of the above-mentioned polysubstituted nitrogen-containing heterocyclic derivative or the above-mentioned fungicide in preventing and controlling plant bacterial diseases.

[0106] In some embodiments, the bacterial diseases include one or more of diseases caused by Ralstonia solanacearum on ginger, diseases caused by Ralstonia solanacearum on tomatoes, diseases caused by Ralstonia solanacearum on peppers, diseases caused by Ralstonia solanacearum on tobacco, diseases caused by Ralstonia solanacearum on potatoes, diseases caused by Ralstonia solanacearum on peanuts, soft rot of fruits and vegetables, ulcer disease, and bacterial blight of rice.

[0107] The following further illustrates the polysubstituted nitrogen-containing heterocyclic derivatives, their preparation methods, fungicides, and applications of the present invention using specific examples. This section further illustrates the present invention with reference to specific examples but should not be construed as limiting the present invention. Unless otherwise specified, the techniques employed in the examples are conventional techniques well known to those skilled in the art. Unless otherwise specified, the reagents, methods, and equipment employed in the present invention are conventional in the art.

[0108] Example 1

[0109] The present invention provides a multi-substituted nitrogen-containing heterocyclic derivative, the structural formula of which is shown below:

[0110]

[0111] Among them, R 1 、R 2 、R 3 、R 4 、R 5 The groups are shown in Table 1 below.

[0112] Table 1 - Substituents of different polysubstituted nitrogen-containing heterocyclic derivatives

[0113]

[0114]

[0115] In Table 1 above, Piperazin-1-yl represents piperazin-1-yl, Cyclopropyl represents C3H5-cyclopropyl, and Propargyl represents propargyl.

[0116] Example 2

[0117] The present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the specific structure of which is shown in Table 1 as compound I2, and the preparation reaction formula thereof is:

[0118]

[0119] The specific preparation method of compound I2 is as follows: (1) adding o-chloroacetophenone (10 mmol, 1 eq) to a round-bottom flask containing 15 mL of THF (tetrahydrofuran), stirring to dissolve it, then adding NaH (12 mmol, 1.2 eq) to the reaction flask under continuous stirring, and stirring at room temperature for 30 minutes; then adding diethyl carbonate (12 mmol, 1.2 eq), heating to reflux for reaction, tracking by TLC until the reaction is complete, quenching the reaction with saturated ammonium chloride, extracting with EA, washing with water, drying, and concentrating to obtain the intermediate o-chlorobenzoyl ethyl acetate; (2) taking the above o-chlorobenzoyl ethyl acetate (6 mmol), dissolving it in 15 mL of toluene, adding DMFDMA (N,N-dimethylformamide dimethyl acetal, 7.2 mmol), then slowly heating to 100 degrees for reaction, and monitoring by TLC until the reaction is complete. Complete, reduce pressure and concentrate to obtain a crude product for use; (3) add 15mL toluene, cyclopropylamine (6mmol), and cesium carbonate (6mmol) to the above crude product in sequence, heat to 130℃, stir and react until the substrate conversion is complete, cool, pour the reaction solution into 25mL of ice water, extract with ethyl acetate, wash with water, dry, and concentrate to obtain the intermediate N-(cyclopropyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid ethyl ester; (4) hydrolyze the intermediate N-(cyclopropyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid ethyl ester obtained in step (3) under alkaline (sodium hydroxide) conditions, complete the reaction, adjust the pH to 2-3 with dilute hydrochloric acid under ice bath conditions, precipitate the solid, and filter to obtain the target product. Analytical data of target compound I2: MS (ESI) m / z 230.3 (M+H) + ,calcd.for C 13 H 11 NO3 m / z=229.1.

[0120] Example 3

[0121] The present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the specific structure of which is shown in Compound I5 in Table 1, and the preparation reaction formula thereof is:

[0122]

[0123] The specific preparation method of compound I5 is as follows: steps (1), (2), and (3) refer to the method of Example 2, except that cyclopropylamine is replaced with α-phenylethylamine in step (3) to prepare the intermediate N-(1-phenylethyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid ethyl ester; (4) the obtained intermediate N-(1-phenylethyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid ethyl ester is hydrolyzed under potassium hydroxide and ethanol. After the reaction is completed, the pH is adjusted to 2-3 with dilute hydrochloric acid in an ice bath, extracted with dichloromethane, washed with water, dried, and concentrated to obtain the target product. Analytical data of target compound I5: MS (ESI) m / z 316.4 (M+Na) + ,calcd.forC 18 H 15 NO3 m / z=293.1.

[0124] Example 4

[0125] The present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the specific structure of which is shown in Compound I6 in Table 1, and the preparation reaction formula thereof is:

[0126]

[0127] The specific preparation method of compound I6 is as follows: steps (1) and (2) refer to the method of Example 2; (3) 15 mL of toluene, p-methoxyaniline (6 mmol), and cesium carbonate (6 mmol) are added to the above crude product in sequence, the temperature is raised to 130° C., and the reaction is stirred until the substrate conversion is complete. The reaction solution is cooled, poured into 25 mL of ice water, extracted with ethyl acetate, washed with water, dried, and concentrated to obtain the intermediate N-(4-methoxyphenyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid ethyl ester; (4) the intermediate N-(4-methoxyphenyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid ethyl ester obtained in step (3) is hydrolyzed under alkaline (sodium hydroxide) conditions. After the reaction is completed, the pH is adjusted to about 2-3 with dilute hydrochloric acid under ice bath conditions, the solid is precipitated, and the target product is obtained by suction filtration. Analytical data of target compound I6: MS (ESI) m / z 296.4 (M+H) + ,calcd.for C 17 H 13 NO4 m / z=295.1.

[0128] Example 5

[0129] The present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the specific structure of which is shown in Compound I10 in Table 1, and the preparation reaction formula thereof is:

[0130]

[0131] The specific preparation method of compound I10 is as follows: steps (1) and (2) refer to the method of Example 2; (3) 15 mL of toluene, p-fluorophenylethylamine (6 mmol), and cesium carbonate (6 mmol) are added to the above crude product in sequence, the temperature is raised to 130° C., and the reaction is stirred until the substrate conversion is complete. The reaction solution is cooled, poured into 25 mL of ice water, extracted with ethyl acetate, washed with water, dried, and concentrated to obtain the intermediate N-(4-fluorophenylethyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid ethyl ester; (4) the intermediate N-(4-fluorophenylethyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid ethyl ester obtained in step (3) is hydrolyzed under sodium hydroxide conditions. After the reaction is completed, the pH is adjusted to 2-3 with dilute hydrochloric acid under ice bath conditions, extracted with dichloromethane, washed with water, dried, and concentrated to obtain the target product. Analytical data of target compound I10: MS (ESI) m / z 312.2 (M+H) + ,calcd.for C 18 H 14 FNO3 m / z = 311.1.

[0132] Example 6

[0133] The present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the specific structure of which is shown in Compound I14 in Table 1, and the preparation reaction formula thereof is:

[0134]

[0135] The specific preparation method of compound I14 is as follows: (1) 10 mmol of o-chloroacetophenone is dissolved in 20 mL of dichloromethane, and 20 mmol of liquid bromine in dichloromethane is added dropwise under ice bath condition. After the addition is completed, the mixture is stirred at room temperature for reaction, and the mixture is monitored by TLC until the reaction is complete. The mixture is washed with water, dried, and concentrated to obtain a bromoacetophenone intermediate; (2) 6 mmol of the bromoacetophenone intermediate is dissolved in 20 mL of anhydrous ethanol, and then 6.6 mmol of sodium benzenesulfinate is added. The mixture is slowly heated to reflux for reaction, and the reaction is monitored by TLC until the reaction is complete. The mixture is concentrated, 20 mL of water is added, and the mixture is extracted with dichloromethane. The mixture is washed with water, dried, and concentrated to obtain 1-(2-chlorophenyl)-2-(phenylsulfonyl)ethyl Ketone intermediate; (3) Take the above 1-(2-chlorophenyl)-2-(phenylsulfonyl)ethanone (5mmol), dissolve it in 15mL of xylene, add DMFDMA (6mmol), then slowly heat to reflux reaction, monitor by TLC until the reaction is complete, and concentrate under reduced pressure to obtain a crude product for use; (4) Add 8-10mL of acetic acid and 5.5mmol of cyclopropylamine to the above crude product, heat to reflux reaction for about 4h, concentrate to dryness, then add 20mL of toluene and 6mmol of cesium carbonate, then heat to reflux reaction, monitor by TLC until the reaction is complete, cool, pour into ice water, extract with dichloromethane, wash with water, dry, concentrate, and separate by column chromatography to obtain the target product. Analytical data of target compound I14: MS (ESI) m / z 326.3 (M+H) + ,calcd.for C 18 H 15 NO3S m / z = 325.1.

[0136] Example 7

[0137] The present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the specific structure of which is shown in Table 1 as compound I24, and the preparation reaction formula thereof is:

[0138]

[0139] The specific preparation method of compound I24 is the same as that of Example 6, except that in step (1), NBS (n-bromosuccinimide) is used as the bromination reagent to prepare the relevant brominated intermediate, and in step (2), sodium benzenesulfinate is replaced with sodium thiophenol. Analytical data of target compound I24: MS (ESI) m / z 295.4 (M+H) + ,calcd.for C 17 H 14 N2OS m / z = 294.1.

[0140] Example 8

[0141] The present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the specific structure of which is shown in Table 1 as compound I27, and the preparation reaction formula thereof is:

[0142]

[0143] The specific preparation method of compound I27 is as follows: steps (1) to (4) refer to the method of Example 2; (5) the intermediate 1-ethyl-7-methyl-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid (1 mmol, 1 eq), HOBt (1.5 mmol, 1.5 eq), EDCI (1.5 mmol, 1.5 eq) and DMF (6 mL) were added in sequence to a 50 mL round-bottom flask and stirred until completely dissolved. Methylsulfonylethylamine hydrochloride (1 mmol, 1 eq) was then added, followed by the slow dropwise addition of Et3N (3 mmol, 3 eq). The reaction was stirred at room temperature overnight and the reaction solution was poured into a beaker containing 30 mL of ice water. Solid precipitated and was filtered and washed with a small amount of water to obtain a gray solid crude product. Analytical data of target compound I27: MS (ESI) m / z 338.2 (M+H) + ,calcd.for C 15 H 19 N3O4S m / z=337.1.

[0144] Example 9

[0145] The present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the specific structure of which is shown in Table 1 as compound I29, and the preparation reaction formula thereof is:

[0146]

[0147] The specific preparation method of compound I29 is as follows: steps (1) to (4) refer to the method of Example 2; (5) the intermediate 1-ethyl-7-methyl-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid (1 mmol, 1 eq) and TBTU (1.5 mmol, 1.5 eq) were added to a 50 mL round-bottom flask, 10 mL of DMF was added and stirred to completely dissolve, Et3N (4 mmol, 4 eq) was added dropwise, and the reaction was stirred for 1 hour. After that, isonicotinic acid hydrazide (1 mmol, 1 eq) was added and stirred at room temperature and monitored by TLC. After the reaction was completed, the reaction solution was cooled to room temperature, 50 mL of water was added and stirred, extracted with DCM, washed with saturated brine, washed with water, dried and rotary evaporated to obtain a crude product, and purified by silica gel column chromatography. Analytical data of target compound I29: MS (ESI) m / z 352.4 (M+H) + ,calcd.for C 18 H 17 N5O3 m / z=351.1.

[0148] Example 10

[0149] The present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the specific structure of which is shown in Table 1 as compound I40, and the preparation reaction formula thereof is:

[0150]

[0151] The specific preparation method of compound I40 is as follows: refer to the method of Example 2, except that the conditions such as different substrates, solvents and reaction temperatures are appropriately replaced according to the experimental conditions. Analytical data of target compound I40: MS (ESI) m / z 283.2 (M+H) + ,calcd.for C 12 H8ClFN2O3 m / z=282.0.

[0152] Example 11

[0153] The present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the specific structure of which is shown in Table 1 as compound I41, and the preparation reaction formula thereof is:

[0154]

[0155] The specific preparation method of compound I41 is as follows: refer to the method of Example 2, except that the conditions such as different substrates, solvents and reaction temperatures are appropriately replaced according to the experimental conditions. Analytical data of target compound I41: MS (ESI) m / z 281.2 (M+H) + ,calcd.for C 12 H6ClFN2O3 m / z=280.0.

[0156] Example 12

[0157] The present invention provides a polysubstituted nitrogen-containing heterocyclic derivative, the specific structure of which is shown in Table 1 as compound I42, and the preparation reaction formula thereof is:

[0158]

[0159] The specific preparation method of compound I42 is as follows: refer to the method of Example 2, except that the conditions such as different substrates, solvents and reaction temperatures are appropriately replaced according to the experimental conditions. Analytical data of target compound I42: MS (ESI) m / z 308.4 (M+H) + ,calcd.for C 13 H7ClFN3O3 m / z=307.0.

[0160] The preparation of other compounds in Table 1 can refer to the basic synthesis methods described in Examples 2-12 above, and different chemical raw materials can be selected in combination with the structural characteristics of the compounds described in Table 1 to prepare the other compounds listed in the table.

[0161] Antibacterial performance test

[0162] The nitrogen-containing heterocyclic compound in Example 1 was selected for preliminary screening of bactericidal activity. The test target was R. solanacearum, and the test method adopted was the Oxford cup method. Specifically, the sterilized agar medium was heated to completely melt, poured into a culture dish, 15 ml per dish (lower layer), and waited for it to solidify. In addition, the melted PDA culture medium was cooled to about 50 ° C and mixed with the test bacteria. 5 ml of the culture medium mixed with bacteria was added to the solidified culture medium to be solidified (upper layer). An Oxford cup was placed vertically directly on the surface of the culture medium with aseptic operation, and pressure was gently applied so that it was in contact with the culture medium without gaps, and the sample to be tested was added to the cup. After addition, the culture medium was placed at 37 ° C with the front side facing up and cultured for 16-18 hours. The results were observed, and the size of the inhibition zone could reflect the activity of the compound.

[0163] Figure 1 The figures show the preliminary screening results of the fungicidal activity of different compounds against Ralstonia solanacearum (only compounds with certain activity are listed, and the compound numbers in the figure correspond to the compound numbers in Example 1). The positive control drugs in the figure are streptomycin and bismethazol.

[0164] from Figure 1 It can be seen that some nitrogen-containing heterocyclic compounds have significant fungicidal activity against Ralstonia solanacearum, especially compounds I2, I36, I38, I40, I42, and I44.

[0165] In order to further investigate the potential inhibitory activity level and application potential of this type of compound against bacterial diseases, the Oxford cup method and turbidimetric method were used to conduct the bactericidal spectrum and LC 50 In the data determination test, the test targets are Escherichia coli (E. coli), soft rot fungus (E. carotovora), and R. solanacearum.

[0166] First, referring to the test method for the bactericidal activity of Ralstonia solanacearum, the Oxford cup method was used to test the bactericidal activity of Escherichia coli ( Figure 2 ), soft rot fungi, solanacearum ( Figure 4 ) were evaluated for their antibacterial activity at different concentrations. The specific evaluation results are as follows Figures 2-4 shown.

[0167] pass Figures 2-4Analysis of the screening results showed that the test compounds had significant fungicidal effects on one or more of the test diseases at the test concentrations; in particular, some compounds had significantly better fungicidal effects than the control drug Streptomycin at a lower test concentration of 200 ppm.

[0168] Subsequently, the bactericidal effect of the above-mentioned highly active compounds was further determined by turbidimetry. The specific test method was referred to the literature [Indoor screening and evaluation of ginger bacterial wilt control agents]. The culture medium without drug solution was used as the negative control, and the culture medium without drug solution but with bacteria was used as the positive control. The absorbance value of each drug solution treatment group was measured by spectrophotometer, and LC was performed using SPSS22.0. 50 The test results of the compounds are shown in Tables 2-4 below (the compound numbers in the table correspond to the compound numbers in Example 1).

[0169] Table 2 Bactericidal activity of compounds against Escherichia coli

[0170] Compound <![CDATA[LC 50 (ppm)]]> Linear equation (logx) <![CDATA[R 2 ]]> I2 0.913 y=0.3881x+0.0149 0.9981 I36 37.917 y=0.5501x-0.8685 0.9984 I38 12.643 y=2.0585x-2.287 0.9625 I40 0.064 y=0.2711x+0.3255 0.9898 I42 6.162 y=0.5485x-0.4324 0.994 I44 0.0000091 y=0.1153x+0.5802 0.964 Streptomycin 1.918 y=0.6656x-0.1886 0.988

[0171] Table 3 Bactericidal activity of compounds against soft rot fungi (E. carotovora)

[0172]

[0173]

[0174] Table 4 Bactericidal activity of compounds against R. solanacearum

[0175] Compound <![CDATA[LC 50 (ppm)]]> Linear equation (logx) <![CDATA[R 2 <!-- 18 -->]]> I2 13.790 y=1.4437x-1.6452 1.000 I36 30.519 y=1.2828x-1.901 0.9633 I38 12.724 y=2.0831x-2.3192 0.9547 I40 0.002 y=0.1808x+0.6009 0.9591 I42 34.008 y=1.1495x-1.7593 0.9658 I44 0.284 y=0.3828x+0.2122 0.9689 Streptomycin 0.169 y=0.4388x+0.3401 0.9965

[0176] The data in Tables 2-4 show that test compounds I2, I40, and I44 exhibited superior activity against Enterobacteriaceae, while compounds I38, I40, and I44 exhibited superior activity against soft rot fungi. Compounds I40 and I44 exhibited significant bactericidal activity against Ralstonia solanacearum. The test results using the Oxford cup and turbidimetric methods demonstrate that these compounds exhibit significant antibacterial activity against bacterial diseases, potentially providing a means to address resistance to existing fungicides and potentially serving as alternatives in production practices. This also provides a novel technical strategy for the prevention and control of current bacterial diseases.

[0177] It can be understood that the fungicide obtained by mixing the polysubstituted nitrogen-containing heterocyclic derivatives of the present application with different adjuvants also has a good fungicidal effect on Escherichia coli, soft rot fungi, solanacearum, etc.

[0178] The above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. Use of a polysubstituted nitrogen-containing heterocyclic derivative or a fungicide in preventing and controlling plant bacterial diseases; the polysubstituted nitrogen-containing heterocyclic derivative is compound I40; The structural formula of the compound I40 is The bactericide includes the multi-substituted nitrogen-containing heterocyclic derivative; The bacterial disease is damage caused by Ralstonia solanacearum.

2. The use according to claim 1, wherein the fungicide further comprises an adjuvant, and the mass fraction of the polysubstituted nitrogen-containing heterocyclic derivative in the fungicide is 0.5 to 99.99%.

3. The use according to claim 2, wherein the auxiliary agent comprises one or more of isopropyl alcohol, glycerol, turpentine, propanol, xylene, chlorobenzene, 1,2-dichloroethane, 1,2-dibromoethane, toluene, methanol, ethanol, N,N-dimethylformamide, ethyl acetate, acetone, butanone, cyclohexanone, dimethyl sulfoxide, and paraffin; Or, the auxiliary agent includes one or more of kaolin, bentonite, clay, diatomaceous earth, montmorillonite, activated clay, dolomite, quartz, calcium carbonate, talc, and attapulgite; Or, the auxiliary agent includes one or more of alkyl sulfonates, alkyl sulfonic acid esters, alkyl aryl sulfonates, sorbitol polyoxyethylene esters, polyoxyethylene-fatty alcohol ethers, polyoxyethylene-fatty acid esters, aralkyl polyethylene glycol ethers, fluoroalkyl sulfonic acid esters, alkyl sulfates, lignin sulfonates, polyethylene glycol, and rhamnolipids; Or, the auxiliary agent includes one or more of polyvinyl alcohol, carboxymethyl cellulose, and gum arabic; Alternatively, the auxiliary agent includes one or more of an inorganic dye, an organic dye and a trace nutrient.

4. The use according to claim 2, characterized in that The fungicide can be prepared into any one of water-dispersible granules, water emulsions, suspensions, wettable powders or nano controlled-release agents.

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