Pyrimido triazole thioether-containing amide derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of chemistry, and particularly relates to a pyrimido triazole thioether-containing amide derivative and a preparation method and application thereof, the structural general formula (I) is shown in the specification, R is selected from one or two of halogen, C1-C4 alkyl and nitryl, the halogen is selected from one or two of fluorine, chlorine, bromine and iodine, and the nitryl is selected from one or two of fluorine, chlorine, bromine and iodine. The pyrimidino triazole thioether-containing amide derivative prepared by the invention has a relatively good inhibition effect on citrus canker pathogens in citrus canker. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of chemistry, and in particular relates to a pyrimidine-triazole thioether amide derivative and a preparation method and application thereof. Background Art

[0002] Due to their unique structural skeleton, pyrimidine and triazole compounds contain two active groups, pyrimidine and triazole. They have unique chemical properties and biological activities and are widely used in many fields, including pesticides and medicines. Compounds with pyrimidotriazole structure have biological activities such as antibacterial (Lin Xuanfu, et al. Organic Chemistry, 2013, 33(02): 353-358.), herbicidal (Xu Li, et al. Chemical Research and Application, 2010, 22(04): 475-478.), anti-inflammatory (Zhang HJ, et al. Bioorganic & Medicinal Chemistry Letters, 2017, 27(18): 4409-4414), anti-tumor (Hassan GS, et al. Arabian Journal of Chemistry, 2017, 10: S1345-S1355.), and antiviral (Fares M, et al. Journal of Molecular Structure, 2022, 1252: 132092.).

[0003] Citrus canker is a widespread disease that can harm dozens of Rutaceae plants and is a major quarantine disease that affects citrus production worldwide. It harms citrus leaves, branches, and fruits. If not treated in time, the disease worsens, affecting not only vegetation growth but also citrus production and economic benefits. Therefore, it is necessary to prevent and control citrus canker. However, the inhibitory activity of existing drugs for the pathogenic bacteria of citrus canker is low. Summary of the invention

[0004] In view of the deficiencies of the above-mentioned prior art, the object of the present invention is to provide a pyrimidine-triazole thioether amide derivative and a preparation method and application thereof. The present invention splices a pyrimidine structure, a 1,2,4-triazole structure and a thioether amide structure to obtain a novel pyrimidine-triazole thioether amide derivative, which can inhibit pathogenic bacteria related to citrus canker.

[0005] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0006] A pyrimidine-triazole thioether amide derivative, the general formula of which is as follows (I):

[0007]

[0008] Wherein, R is selected from one or two of halogen, C1-C4 alkyl and nitro.

[0009] In a preferred embodiment of the present invention, the halogen is selected from one or two of fluorine, chlorine, bromine and iodine.

[0010] In a preferred embodiment of the present invention, the position of the halogen, alkyl or nitro substituent in R may be ortho, ortho, para or meta substitution.

[0011] In a preferred embodiment of the present invention, the pyrimidine-triazole thioether amide derivatives can be specifically selected from the following compounds:

[0012] N-(3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0013] N-(4-chloro-3-fluorophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0014] N-(4-chloro-3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0015] N-(3-chloro-4-fluorophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0016] N-(4-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0017] N-(4-bromophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0018] N-(m-Tolyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0019] N-(2-chloro-4-iodophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0020] N-(2,3-dimethylphenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0021] N-(4-ethylphenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0022] N-(4-butylphenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0023] N-(2-Fluoro-phenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0024] N-(4-methyl-3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0025] N-(2-chloro-4-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0026] N-(4-fluoro-3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0027] N-(4-fluoro-2-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0028] N-(2-methyl-4-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0029] N-(2-iodophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0030] N-(o-Tolyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0031] N-(2-bromophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide;

[0032] N-(4-iodophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide.

[0033] In a preferred embodiment of the present invention, R is one of 3-nitrophenyl, 4-chloro-3-fluorophenyl, 4-chloro-3-nitrophenyl, 3-chloro-4-fluorophenyl, 4-nitrophenyl, 4-bromophenyl, 3-methylphenyl, 2-chloro-4-iodophenyl, 2,3-dimethylphenyl, 4-ethylphenyl, 4-butylphenyl, 2-fluorophenyl, 4-methyl-3-nitrophenyl, 2-chloro-4-nitrophenyl, 4-fluoro-3-nitrophenyl, 4-fluoro-2-nitrophenyl, 2-methyl-4-nitrophenyl, 2-iodophenyl, 2-methylphenyl, 2-bromophenyl, and 4-iodophenyl.

[0034] Another object of the present invention is to provide a method for preparing the above-mentioned pyrimidotriazole sulfide amide derivatives, comprising the following steps:

[0035] The product was synthesized by the following method using ethyl trifluoroacetoacetate, formamidine hydrochloride, hydrazine hydrate, carbon disulfide and ethyl bromoacetate as raw materials.

[0036] Synthesis of the compound shown in formula 1:

[0037]

[0038] Add methanol to the bottle, weigh metal Na (0.3 mol), wipe dry, cut into small pieces and slowly add to methanol. Stir the reaction at room temperature, add the corresponding proportion of formamidine hydrochloride (0.2 mol) after 1 hour, and stir at room temperature for half an hour. Finally, add ethyl trifluoroacetoacetate (0.22 mol) to react for 24 hours, and monitor the reaction by TLC. After the reaction is completed, remove the solution to obtain a milky white mixture, cool it, add water to completely dissolve the solid. Subsequently, use 15% dilute hydrochloric acid solution to adjust the pH of the reaction system to 2. A white solid precipitates. Subsequently, filter and dry to obtain hydroxypyrimidine.

[0039] Weigh the above solid (0.1 mol) in a three-necked flask, use acetonitrile as solvent, measure N, N-diisopropylethylamine (0.12 mol) and add it dropwise to the reaction solution under ice bath, and stir for 10 minutes. Then measure phosphorus oxychloride (0.2 mol), and slowly add the reagent dropwise to the reactant using a constant temperature dropping funnel. After the addition is complete, continue stirring for 10 minutes under ice bath conditions. Subsequently, heat reflux at 85°C for 4-6 hours. TLC monitors the progress of the reaction. After the reaction is completed, cool to room temperature, and then decompress and spin dry the solvent to obtain a mixture. Under ice bath conditions, slowly add ice water to the mixture, and then add saturated sodium bicarbonate solution to adjust the pH value to 6-7. Subsequently, extract with dichloromethane 2-3 times. After that, dry it with anhydrous sodium sulfate to remove residual moisture. Spin dry the solvent to obtain a light yellow oily product to obtain the compound shown in Formula 1.

[0040] Synthesis of the compound shown in formula 2:

[0041]

[0042] The compound shown in Formula 1 and hydrazine hydrate are used as raw materials and isopropanol is used as solvent. A condensation reaction occurs under ice bath conditions, and the compound shown in Formula 2 is obtained through post-treatment.

[0043] Synthesis of the compound shown in formula 3:

[0044]

[0045] In anhydrous ethanol solvent, 4-hydrazino-6-trifluoromethylpyrimidine compound represented by formula 2 and carbon disulfide are used as raw materials, potassium hydroxide is added thereto as a condensation agent, and the mixture is heated under reflux to undergo a condensation reaction. After post-treatment, a compound represented by formula 3, i.e., 7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidine-2-thiol, is obtained.

[0046] Synthesis of compound 4:

[0047]

[0048] In anhydrous ethanol solvent, triethylamine and tetrabutylammonium iodide are added to the compound of formula 3 and ethyl bromoacetate as raw materials to cause a condensation reaction. After post-treatment, a compound of formula 4, i.e., ethyl 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetate, is obtained.

[0049] Synthesis of the compound shown in Formula 5:

[0050]

[0051] In tetrahydrofuran solvent, potassium hydroxide and water are added to the compound of formula 4 as a raw material, and a condensation reaction occurs under ice bath conditions. After post-treatment, a compound of formula 5 is obtained, namely 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetic acid.

[0052] Synthesis of Compound I:

[0053]

[0054] In a dichloromethane solvent, the compound represented by formula 5 and aniline with different substituents are used as raw materials, 1-hydroxybenzotriazole (HOBT) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) are added thereto to carry out a condensation reaction, and after post-treatment, compound (I), i.e., a pyrimidine-triazole thioether amide derivative, is obtained, wherein R is selected from one or two of halogen, C1-C4 alkyl and nitro.

[0055]

[0056] In a preferred embodiment of the present invention, in the process of preparing the compound represented by formula 2, the molar ratio of the compound represented by formula 1 and hydrazine hydrate is 0.02:0.03, and in the process of preparing the compound represented by formula 3, the molar ratio of 4-hydrazino-6-trifluoromethylpyrimidine, carbon disulfide and potassium hydroxide is 1:1~2:1~2.

[0057] In a preferred embodiment of the present invention, in the process of preparing the compound of formula 4, the molar ratio of the compound of formula 3, ethyl bromoacetate, triethylamine and tetrabutylammonium iodide is 3:3.6-4:7-8:0.01.

[0058] In a preferred embodiment of the present invention, during the preparation of the compound represented by Formula 5, the molar ratio of the compound represented by Formula 4 to potassium hydroxide is 1:2-4.

[0059] In a preferred embodiment of the present invention, during the preparation of compound (I), the molar ratio of the compound represented by formula 5, 1-hydroxybenzotriazole (HOBT), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI) and aniline with different substituents is 1:1.5~2:1.5~2:1.5~2.

[0060] In a preferred embodiment of the present invention, the reaction temperature for preparing the compound represented by formula 3 is 80°C and the reaction time is 4 to 6 hours, the reaction temperature for preparing the compound represented by formula 4 is 78 to 85°C and the reaction time is 6 to 8 hours, and the reaction temperature for preparing the compound represented by formula 5 is -10 to 10°C and the reaction time is 3 to 5 hours.

[0061] The third object of the present invention is to provide a use of the above-mentioned pyrimidine-triazole thioether amide derivatives and their pesticide-acceptable salts in the preparation of drugs for inhibiting crop-related pathogens.

[0062] The present invention also provides an application of the above-mentioned pyrimidine-triazole thioether amide derivatives and their pesticide-acceptable salts in the preparation of drugs for inhibiting citrus canker bacteria.

[0063] Compared with the prior art, the present invention has the following beneficial effects:

[0064] 1. The present invention splices a pyrimidine structure, a 1,2,4-triazole structure and a thioetheramide structure to obtain a novel pyrimidine-triazole thioetheramide derivative, which can inhibit pathogenic bacteria related to citrus canker.

[0065] 2. The biological activity test results of the pyrimidine-triazole thioether amide derivatives provided by the present invention show that the novel pyrimidine-triazole thioether amide derivatives prepared by the present invention have a good inhibitory effect on citrus canker pathogens in citrus canker.

[0066] 3. The synthesis method of the pyrimidine-containing triazole derivatives of the present invention is simple, the conditions are mild, and it is suitable for industrial production. DETAILED DESCRIPTION

[0067] The following is a detailed description of the technical solutions in the embodiments of the present invention in combination with preferred embodiments and data. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0068] It should be noted that all professional terms used in the present invention are only for the purpose of describing specific embodiments and are not intended to limit the scope of protection of the present invention. Unless otherwise specifically stated, various raw materials, reagents, instruments and equipment used in the following embodiments of the present invention can be purchased from the market or prepared by existing methods.

[0069] It is synthesized by the following method using ethyl trifluoroacetoacetate, formamidine hydrochloride, hydrazine hydrate, carbon disulfide and ethyl bromoacetate as raw materials:

[0070] Synthesis of compound 1:

[0071]

[0072] Add methanol to the bottle, weigh metal Na (0.3 mol), wipe dry, cut into small pieces and slowly add to methanol. Stir the reaction at room temperature, add the corresponding proportion of formamidine hydrochloride (0.2 mol) after 1 hour, and stir at room temperature for half an hour. Finally, add ethyl trifluoroacetoacetate (0.22 mol) to react for 24 hours, and monitor the reaction by TLC. After the reaction is completed, remove the solution to obtain a milky white mixture, cool it, add water to completely dissolve the solid. Subsequently, use 15% dilute hydrochloric acid solution to adjust the pH of the reaction system to 2. A white solid precipitates. Subsequently, filter and dry to obtain hydroxypyrimidine.

[0073] Weigh the above solid (0.1 mol) in a three-necked flask, use acetonitrile as solvent, measure N, N-diisopropylethylamine (0.12 mol) and add it dropwise to the reaction solution under ice bath, and stir for 10 minutes. Then measure phosphorus oxychloride (0.2 mol), and slowly add the reagent dropwise to the reactant using a constant temperature dropping funnel. After the addition is complete, continue stirring for 10 minutes under ice bath conditions. Subsequently, heat reflux at 85°C for 4-6 hours. TLC monitors the progress of the reaction. After the reaction is completed, cool to room temperature, and then reduce the pressure to spin dry the solvent to obtain a mixture. Under ice bath conditions, ice water is slowly added to the mixture, and then a saturated sodium bicarbonate solution is added to adjust the pH value to 6-7. Subsequently, dichloromethane is used for extraction 2-3 times. After that, it is dried over anhydrous sodium sulfate to remove residual moisture. The solvent is spin-dried to obtain a light yellow oily product to obtain compound 1.

[0074] Synthesis of compound 2:

[0075]

[0076] Hydrazine hydrate (0.03 mol) was added to a round-bottomed flask, and isopropanol was used as the solvent. After stirring at room temperature for 30 min, a mixture of compound 1 (0.02 mol) and isopropanol was added dropwise to the reaction system under an ice bath using a constant pressure dropping funnel. After completion of the dropwise addition, stirring reaction was continued under an ice bath, and the progress was monitored by TLC during the reaction. After the reaction was completed, the solvent was removed to obtain a light yellow solid, and then water was added to dissolve the solid, and ethyl acetate was used for extraction 2-3 times, and anhydrous sodium sulfate was dried to remove the moisture in the organic phase. The solvent was spin-dried to obtain a white solid to obtain compound 2.

[0077] Example 1

[0078] The synthesis method of N-(3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ia) comprises the following steps:

[0079] (1) Synthesis of 7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidine-2-thiol

[0080] 4-Hydrazino-6-trifluoromethylpyrimidine (0.05 mol) and potassium hydroxide (0.075 mol) were added to a three-necked flask, and anhydrous ethanol was used as the solvent. Carbon disulfide (0.075 mol) was added under stirring, and the mixture was heated under reflux at 80°C for 4-6 hours. After the reaction was completed, the solvent was dried, and a small amount of water was added to dissolve the solid under stirring at room temperature. The pH of the solution was adjusted to 2 with a 10% dilute hydrochloric acid solution. Solids were precipitated, filtered, and dried to obtain a yellow solid with a yield of 82.36%.

[0081] (2) Synthesis of ethyl 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetate:

[0082] 7-(Trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidine-2-thiol (3mmol) and triethylamine (1.08mL) were added to a round-bottom flask, and anhydrous ethanol was used as the solvent. After reflux reaction for 15 minutes, a mixed solution of TBAI (0.01mmol) and ethyl bromoacetate (3.6mmol) dissolved in anhydrous ethanol was added dropwise, and the reflux reaction was continued for 6-8 hours. After the reaction was completed, the solvent was removed, water was added and stirred at room temperature to precipitate a white solid, which was filtered and dried to obtain a white solid with a yield of 84.50%.

[0083] (3) Synthesis of 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetic acid:

[0084] Ethyl 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetate (0.1mmol) was added to a round-bottom flask, and tetrahydrofuran was used as the solvent. After cooling for 10 minutes under ice bath conditions, potassium hydroxide (0.2mmol) solution was added while stirring. The ice bath was always maintained during the reaction. After the reaction was completed, the solvent was dried as much as possible, the mixture was cooled to room temperature, and then an appropriate amount of water was added thereto. The mixture gradually dissolved and became clear, and then glacial acetic acid was used to adjust the pH until solids precipitated. A white solid was obtained by suction filtration and drying, with a yield of 42.37%.

[0085] (4) Synthesis of N-(3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide

[0086] 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetic acid (0.1mmol), HOBT (0.15mmol) and EDCI (0.15mmol) were added to a flask, dichloromethane was used as solvent, and the reaction was stirred at room temperature for 30 minutes, and then 3-nitroaniline (0.15mmol) was added dropwise. After the reaction was completed, 10% dilute hydrochloric acid solution, saturated sodium bicarbonate solution, saturated brine and pure water were used to wash the reaction, and the organic phase was dried over anhydrous sodium sulfate, and then the solvent was dried to obtain a crude product, and finally the target compound was purified by column chromatography as a white solid with a yield of 43.67% and mp 156.6-158.8°C.

[0087] Example 2

[0088] The synthesis method of N-(4-chloro-3-fluorophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ib) comprises the following steps:

[0089] (1) Synthesis of 7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidine-2-thiol

[0090] 4-Hydrazino-6-trifluoromethylpyrimidine (0.05 mol) and potassium hydroxide (0.075 mol) were added to a three-necked flask, and anhydrous ethanol was used as the solvent. Carbon disulfide (0.075 mol) was added under stirring, and the mixture was heated under reflux at 80°C for 4-6 hours. After the reaction was completed, the solvent was dried, and a small amount of water was added to dissolve the solid under stirring at room temperature. The pH of the solution was adjusted to 2 with a 10% dilute hydrochloric acid solution. Solids were precipitated, filtered, and dried to obtain a yellow solid with a yield of 82.36%.

[0091] (2) Synthesis of ethyl 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetate:

[0092] 7-(Trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidine-2-thiol (3mmol) and triethylamine (1.08mL) were added to a round-bottom flask, and anhydrous ethanol was used as the solvent. After reflux reaction for 15 minutes, a mixed solution of TBAI (0.01mmol) and ethyl bromoacetate (3.6mmol) dissolved in anhydrous ethanol was added dropwise, and the reflux reaction was continued for 6-8 hours. After the reaction was completed, the solvent was removed, water was added and stirred at room temperature to precipitate a white solid, which was filtered and dried to obtain a white solid with a yield of 84.50%.

[0093] (3) Synthesis of 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetic acid:

[0094] Ethyl 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetate (0.1mmol) was added to a round-bottom flask, and tetrahydrofuran was used as the solvent. After cooling for 10 minutes under ice bath conditions, potassium hydroxide (0.2mmol) solution was added while stirring. The ice bath was always maintained during the reaction. After the reaction was completed, the solvent was dried as much as possible, the mixture was cooled to room temperature, and then an appropriate amount of water was added thereto. The mixture gradually dissolved and became clear, and then glacial acetic acid was used to adjust the pH until solids precipitated. A white solid was obtained by suction filtration and drying, with a yield of 42.37%.

[0095] (4) Synthesis of N-(3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide

[0096] 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetic acid (0.1mmol), HOBT (0.15mmol) and EDCI (0.15mmol) were added to a flask, dichloromethane was used as solvent, and the reaction was stirred at room temperature for 30 minutes, and then 4-chloro-3-fluoroaniline (0.15mmol) was added dropwise. After the reaction was completed, 10% dilute hydrochloric acid solution, saturated sodium bicarbonate solution, saturated brine and pure water were used to wash the reaction, and the organic phase was dried over anhydrous sodium sulfate, and then the solvent was dried to obtain a crude product, and finally the target compound was purified by column chromatography as a white solid with a yield of 28.46% and mp 180.4-182.2°C.

[0097] Example 3

[0098] The synthesis method of N-(4-chloro-3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ic) comprises the following steps:

[0099] (1) Synthesis of 7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidine-2-thiol

[0100] 4-Hydrazino-6-trifluoromethylpyrimidine (0.05 mol) and potassium hydroxide (0.075 mol) were added to a three-necked flask, and anhydrous ethanol was used as the solvent. Carbon disulfide (0.075 mol) was added under stirring, and the mixture was heated under reflux at 80°C for 4-6 hours. After the reaction was completed, the solvent was dried, and a small amount of water was added to dissolve the solid under stirring at room temperature. The pH of the solution was adjusted to 2 with a 10% dilute hydrochloric acid solution. Solids were precipitated, filtered, and dried to obtain a yellow solid with a yield of 82.36%.

[0101] (2) Synthesis of ethyl 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetate:

[0102] 7-(Trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidine-2-thiol (3mmol) and triethylamine (1.08mL) were added to a round-bottom flask, and anhydrous ethanol was used as the solvent. After reflux reaction for 15 minutes, a mixed solution of TBAI (0.01mmol) and ethyl bromoacetate (3.6mmol) dissolved in anhydrous ethanol was added dropwise, and the reflux reaction was continued for 6-8 hours. After the reaction was completed, the solvent was removed, water was added and stirred at room temperature to precipitate a white solid, which was filtered and dried to obtain a white solid with a yield of 84.50%.

[0103] (3) Synthesis of 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetic acid:

[0104] Ethyl 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetate (0.1mmol) was added to a round-bottom flask, and tetrahydrofuran was used as the solvent. After cooling for 10 minutes under ice bath conditions, potassium hydroxide (0.2mmol) solution was added while stirring. The ice bath was always maintained during the reaction. After the reaction was completed, the solvent was dried as much as possible, the mixture was cooled to room temperature, and then an appropriate amount of water was added thereto. The mixture gradually dissolved and became clear, and then glacial acetic acid was used to adjust the pH until solids precipitated. A white solid was obtained by suction filtration and drying, with a yield of 42.37%.

[0105] (4) Synthesis of N-(3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide

[0106] 2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetic acid (0.1mmol), HOBT (0.15mmol) and EDCI (0.15mmol) were added to a flask, dichloromethane was used as solvent, and the reaction was stirred at room temperature for 30 minutes, and then 4-chloro-3-nitroaniline (0.15mmol) was added dropwise. After the reaction was completed, 10% dilute hydrochloric acid solution, saturated sodium bicarbonate solution, saturated brine and pure water were used to wash the reaction, and the organic phase was dried over anhydrous sodium sulfate, and then the solvent was dried to obtain a crude product, and finally the target compound was purified by column chromatography as a white solid with a yield of 46.15% and mp 126.3-128.2°C.

[0107] Example 4

[0108] The synthesis method of N-(3-chloro-4-fluorophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Id) is the same as that of Example 1, except that the raw material added in step (4) is 3-chloro-4-fluoroaniline.

[0109] Example 5

[0110] The synthesis method of N-(4-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ie) is the same as that of Example 1, except that the raw material added in step (4) is 4-nitroaniline.

[0111] Example 6

[0112] The synthesis method of N-(4-bromophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (If) is the same as that of Example 1, except that the raw material added in step (4) is 4-bromoaniline.

[0113] Example 7

[0114] The synthesis method of N-(m-tolyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ig) is the same as that in Example 1, except that the raw material added in step (4) is m-toluidine.

[0115] Example 8

[0116] The synthesis method of N-(2-chloro-4-iodophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ig) is the same as that in Example 1, except that the raw material added in step (4) is 2-chloro-4-iodoaniline.

[0117] Example 9

[0118] The synthesis method of N-(2-chloro-4-iodophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ih) is the same as that in Example 1, except that the raw material added in step (4) is 2-chloro-4-iodoaniline.

[0119] Example 10

[0120] The synthesis method of N-(2,3-dimethylphenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ii) is the same as that in Example 1, except that the raw material added in step (4) is 2,3-dimethylaniline.

[0121] Embodiment 11

[0122] The synthesis method of N-(4-ethylphenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ij) is the same as that in Example 1, except that the raw material added in step (4) is 4-ethylphenylamine.

[0123] Example 12

[0124] The synthesis method of N-(4-butylphenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ik) is the same as that in Example 1, except that the raw material added in step (4) is 4-butylaniline.

[0125] Example 13

[0126] The synthesis method of N-(2-fluoro-phenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Il) is the same as that in Example 1, except that the raw material added in step (4) is 2-fluoro-aniline.

[0127] Embodiment 14

[0128] The synthesis method of N-(4-methyl-3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Im) is the same as that in Example 1, except that the raw material added in step (4) is 4-methyl-3-nitroaniline.

[0129] Embodiment 15

[0130] The synthesis method of N-(2-chloro-4-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (In) is the same as that in Example 1, except that the raw material added in step (4) is 2-chloro-4-nitroaniline.

[0131] Example 16

[0132] The synthesis method of N-(2-chloro-4-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (In) is the same as that in Example 1, except that the raw material added in step (4) is 2-chloro-4-nitroaniline.

[0133] Embodiment 17

[0134] The synthesis method of N-(4-fluoro-3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Io) is the same as that in Example 1, except that the raw material added in step (4) is 4-fluoro-3-nitroaniline.

[0135] Embodiment 18

[0136] The synthesis method of N-(4-fluoro-2-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ip) is the same as that in Example 1, except that the raw material added in step (4) is 4-fluoro-2-nitrophenylamine.

[0137] Embodiment 19

[0138] The synthesis method of N-(2-methyl-4-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Iq) is the same as that in Example 1, except that the raw material added in step (4) is 2-methyl-4-nitroaniline.

[0139] Embodiment 20

[0140] The synthesis method of N-(2-iodophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Ir) is the same as that in Example 1, except that the raw material added in step (4) is 2-iodophenylamine.

[0141] Embodiment 21

[0142] The synthesis method of N-(o-tolyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Is) is the same as that in Example 1, except that the raw material added in step (4) is o-toluidine.

[0143] Embodiment 22

[0144] The synthesis method of N-(2-bromophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (It) is the same as that in Example 1, except that the raw material added in step (4) is 2-bromophenylamine.

[0145] Embodiment 23

[0146] The synthesis method of N-(4-iodophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide (Iu) is the same as that in Example 1, except that the raw material added in step (4) is 4-iodophenyl.

[0147] Results Characterization

[0148] The H NMR spectra of the synthesized pyrimidine-triazole sulfide amide derivatives ( 1 H NMR) is shown in Table 1; C NMR spectrum (13 C NMR) data are shown in Table 2; physicochemical properties and elemental analysis data are shown in Table 3.

[0149] Table 1 H NMR spectrum data of target compounds

[0150]

[0151]

[0152]

[0153]

[0154] Table 2 C NMR spectrum data of target compounds

[0155]

[0156]

[0157]

[0158] Table 3 Physicochemical properties and elemental analysis of target compounds

[0159]

[0160]

[0161] Test example: Inhibitory activity of the target compound against Xanthomonas axonopodispv. citri (Xac), a pathogen of citrus canker.

[0162] (1) Test method

[0163] The turbidity method was used to culture bacteria in LB broth to determine the inhibitory activity of the target compound against the test bacteria.

[0164] Activation of bacteria: Inoculate the bacterial strain on LB nutrient agar medium and culture it in a 28°C incubator for 3-5 days. Then select a single colony from the activated medium and inoculate it into a conical flask containing liquid culture medium. Then place it in a constant temperature shaker for shaking culture. Until the bacteria grow to the logarithmic growth phase, the number of bacteria increases exponentially and the activity reaches the best state.

[0165] Preparation of test solution: 0.00375 g of the test compound was dissolved in 150 μL of DMSO solution to prepare a high-concentration stock solution, and then different volumes of stock solution were added to 5 mL centrifuge tubes, specifically 80 μL and 40 μL, and finally the volume was fixed to 4 mL with 0.1% Tween aqueous solution.

[0166] Activity test and result calculation: Take 1mL of the solution to be tested and add it to a test tube containing 4mL of sterilized LB medium, that is, a drug-containing culture medium with a test concentration of 100μg / mL and 50μg / mL is prepared. At the same time, the commercial drugs thiophanate-methyl, thiophanate-methyl and thiophanate-zinc are treated in the same way to obtain a positive control group, and an equal amount of DMSO solution is added to the culture medium as a negative control group. Then, 40μL of bacterial solution is added to the above-mentioned treatment group, and then placed in a constant temperature shaker at a temperature of 28°C and a speed of 180rpm for shaking culture for about 24-48 hours, until the bacteria in the negative control group grow to the logarithmic phase. Then, aspirate 200μL of the bacterial solution in the test tube, and then use an enzyme reader to measure the absorbance of the bacterial solution at 595nm, and the inhibition rate is calculated according to the following formula:

[0167] Inhibition rate

[0168] Where: A0: OD value of negative control group;

[0169] A t : OD value of the drug solution group;

[0170] I: inhibition rate.

[0171] (2) Bioassay results

[0172] Table 4 Inhibitory activity of target compounds against citrus canker pathogens in citrus canker

[0173]

[0174]

[0175] From the experimental data in Table 4, it can be seen that the pyrimidine triazole derivatives containing amide structure all showed certain inhibitory activity against citrus canker bacteria in citrus canker disease. Almost all the compounds had good inhibitory activity against citrus canker bacteria, and the inhibition rate of compounds c, d, g, h, i, j, k, m, p, r, and u was 100% at 100 μg / mL, while compounds c, h, j, k, and u still had 100% inhibitory effect at 50 μg / mL, and the inhibitory effect was higher than that of the positive control drug thiophanate-copper.

[0176] (3) EC of some compounds against Xanthomonas citri citri canker 50 value

[0177] Table 5 EC values ​​of some compounds against Xanthomonas citri citri canker 50 value

[0178]

[0179]

[0180] From the experimental data in Table 5, it can be seen that the EC 50 The values ​​were higher than those of the positive control drug thiophanate-methyl, among which the EC values ​​of compounds h, j, m, and u were 50 The values ​​were 3.39, 8.56, 6.97 and 1.56 μg / mL respectively.

[0181] In summary, the novel pyrimidine-triazole thioether amide derivatives prepared by the present invention have a good inhibitory effect on citrus canker pathogens in citrus canker.

[0182] It should be noted that when the present invention involves a numerical range, it should be understood that the two endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as those in the embodiments, in order to avoid redundancy, the present invention describes a preferred embodiment. Although the preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they know the basic creative concept. Therefore, the attached claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0183] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A pyrimidotriazole thioether amide derivative, characterized in that: Its general formula is as follows: Wherein, R is selected from one or two of halogen, C1-C4 alkyl and nitro.

2. The pyrimidotriazole thioether amide derivative according to claim 1, characterized in that: The halogen is selected from one or two of fluorine, chlorine, bromine and iodine.

3. The pyrimidotriazole thioether amide derivative according to claim 1, characterized in that: The pyrimidine-triazole thioether amide derivatives are specifically selected from the following compounds: N-(3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(4-chloro-3-fluorophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(4-chloro-3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(3-chloro-4-fluorophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(4-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(4-bromophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(m-Tolyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(2-chloro-4-iodophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(2,3-dimethylphenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(4-ethylphenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(4-butylphenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(2-Fluorophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(4-methyl-3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(2-chloro-4-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(4-fluoro-3-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(4-fluoro-2-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(2-methyl-4-nitrophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(2-iodophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(o-Tolyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(2-bromophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide; N-(4-iodophenyl)-2-((7-(trifluoromethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)thio)acetamide.

4. A method for preparing the pyrimidotriazole sulfide amide derivative according to claim 1, characterized in that: The following steps are involved: The compound represented by Formula 1 and hydrazine hydrate are used as raw materials, isopropanol is used as solvent, and a condensation reaction is carried out under ice bath conditions, and after post-treatment, a compound represented by Formula 2 is obtained; In anhydrous ethanol solvent, the compound represented by formula 2 and carbon disulfide are used as raw materials, potassium hydroxide is added thereto as a condensation agent, and the mixture is heated under reflux to undergo a condensation reaction, and after post-treatment, a compound represented by formula 3 is obtained; In anhydrous ethanol solvent, the compound represented by formula 3 and ethyl bromoacetate are used as raw materials, triethylamine and tetrabutylammonium iodide are added thereto, a condensation reaction occurs, and after post-treatment, a compound represented by formula 4 is obtained; In tetrahydrofuran solvent, the compound represented by formula 4 is used as a raw material, potassium hydroxide and water are added thereto, and a condensation reaction occurs under ice bath conditions, and after post-treatment, a compound represented by formula 5 is obtained; In a dichloromethane solvent, the compound represented by Formula 5 and the compound represented by Formula 6 are used as raw materials, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride are added thereto, a condensation reaction occurs, and after post-treatment, a compound represented by Formula I is obtained; The synthetic route is as follows: Wherein, R is selected from one or two of halogen, C1-C4 alkyl and nitro.

5. The method for preparing a pyrimidotriazole sulfide amide derivative according to claim 4, characterized in that: In the process of preparing the compound shown in formula 2, the molar ratio of the compound shown in formula 1 and hydrazine hydrate is 0.02:0.

03. In the process of preparing the compound shown in formula 3, the molar ratio of 4-hydrazino-6-trifluoromethylpyrimidine, carbon disulfide and potassium hydroxide is 1:1~2:1~2.

6. The method for preparing a pyrimidotriazole sulfide amide derivative according to claim 4, characterized in that: In the process of preparing the compound represented by formula 4, the molar ratio of the compound represented by formula 3, ethyl bromoacetate, triethylamine and tetrabutylammonium iodide is 3:3.6-4:7-8:0.

01.

7. The method for preparing a pyrimidotriazole sulfide amide derivative according to claim 4, characterized in that: In the process of preparing the compound represented by formula 5, the molar ratio of the compound represented by formula 4 to potassium hydroxide is 1:2-4.

8. The method for preparing a pyrimidotriazole thioether amide derivative according to claim 4, characterized in that: In the process of preparing the compound represented by formula I, the molar ratio of the compound represented by formula 5, 1-hydroxybenzotriazole, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide and aniline with different substituents is 1:1.5~2:1.5~2:1.5~2.

9. The method for preparing a pyrimidotriazole sulfide amide derivative according to claim 4, characterized in that: The reaction temperature for preparing the compound represented by formula 3 is 80°C and the reaction time is 4 to 6 hours. The reaction temperature for preparing the compound represented by formula 4 is 78 to 85°C and the reaction time is 6 to 8 hours. The reaction temperature for preparing the compound represented by formula 5 is -10 to 10°C and the reaction time is 3 to 5 hours.

10. Use of the pyrimidine-triazole thioether amide derivatives and their pesticide-acceptable salts as claimed in claim 1 in the preparation of drugs for inhibiting citrus canker bacteria.

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