A propiolate antibacterial agent and its preparation method and application

Abstract

The present invention belongs to the field of medicinal chemistry research, and specifically discloses a propiolate antibacterial agent and a preparation method and application thereof. The propiolate compound has a general structural formula as shown in the following general formulas I and II. The compound provided by the present invention has different degrees of inhibitory effects on bacteria or fungi related to agricultural and food pollution, especially has better antibacterial activity against methicillin-resistant Staphylococcus aureus, which also provides a lead compound for the development of drugs for preventing and treating bacteria or fungi related to agricultural and food pollution, especially for the prevention or treatment of methicillin-resistant Staphylococcus aureus, and has a good development prospect.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry research, and specifically relates to a propiolate antibacterial agent, a pharmaceutical composition containing the compound, a preparation method, and the use of the compounds in preparing drugs for preventing and treating bacteria or fungi associated with agricultural and food pollution, especially in preparing drugs for preventing or treating methicillin-resistant Staphylococcus aureus. Background Art

[0002] The agricultural and food industries are often contaminated by bacteria or fungi, which affects the quality of agricultural products and food quality safety, poses a threat to human health, and can cause death in severe cases. Staphylococcus aureus is a Gram-positive bacterium belonging to the genus Staphylococcus. It has a certain tolerance to high temperatures and can survive in high-salt environments. Staphylococcus aureus is widely present in the air, soil, and healthy humans and animals. It often contaminates food through raw materials and is one of the main causes of foodborne bacterial poisoning worldwide. In particular, food poisoning caused by Staphylococcus aureus ranks second in global food poisoning incidents. Foodborne diseases caused by Staphylococcus aureus contamination account for 20-25% of foodborne diseases in my country. Staphylococcus aureus can acquire antibiotic resistance determinants, so Staphylococcus aureus isolates often show resistance to multiple antimicrobial drugs. Among them, methicillin-resistant Staphylococcus aureus acquires resistance to penicillin and other antibiotics (such as β-lactams, tetracyclines, aminoglycosides, macrolides, etc.) by carrying the mecA gene encoding penicillin-binding protein 2a (PBP2a). In addition, MRSA has the ability to colonize and infect humans and animals, most notably causing infections of the skin, soft tissue, bones, and lower respiratory tract. It is one of the important multidrug-resistant pathogens that cause hospital, community, and livestock infections, resulting in considerable morbidity and mortality, and is considered a serious global health problem that needs to be addressed urgently. Currently, the main antibiotics for the treatment of MRSA are vancomycin and some newer drugs, such as linezolid, tigecycline, daptomycin, telavancin, ceftaroline, and a combination of quinupristin and dalfopristin. However, MRSA strains resistant to these drugs have been reported one after another, so it is urgent to develop new candidate drugs to prevent or treat MRSA infections. Summary of the invention

[0003] In order to find drugs with high antibacterial activity, the inventors of the present invention have designed and synthesized a series of novel propiolate derivatives with novel structures and excellent antibacterial activity against a variety of bacteria or fungi, especially methicillin-resistant Staphylococcus aureus, after extensive and in-depth research, and studied the antibacterial activity of this type of novel derivatives against common bacteria and fungi in the agriculture or food industry.

[0004] The primary purpose of the present invention is to provide a propiolate compound, and its stereoisomers, prodrugs, and pharmaceutically acceptable salts or pharmaceutically acceptable solvates;

[0005] Another object of the present invention is to provide a method for preparing the above compound;

[0006] Another object of the present invention is to provide a pharmaceutical composition comprising the above compound;

[0007] Another object of the present invention is to provide the use of the above compound in preparing a drug for preventing and treating bacteria or fungi associated with agricultural and food pollution, especially in preparing a drug for preventing or treating methicillin-resistant Staphylococcus aureus.

[0008] The present invention is achieved through the following technical methods.

[0009] A propiolate compound, the general structural formula of which is shown in the following general formulas I and II:

[0010]

[0011] in:

[0012] A in general formula I and II can be an element such as carbon, oxygen, sulfur, selenium or tellurium;

[0013] In the general formula II, n is 1, 2, 3 or 4;

[0014] R in Formula I and II 1 Selected from:

[0015] 1) N,N-dialkylaminothioformyl, N-alkylaminothioformyl, N,N-dialkylcarbamoyl or N-alkylcarbamoyl, wherein the alkyl group mentioned above refers to a C1-C12 straight chain or branched chain alkyl group.

[0016] 2) Wherein Z1, Z2, Z3, Z4, and Z5 are each independently selected from:

[0017] (1) Hydrogen, fluorine, chlorine, bromine, iodine, methyl, benzyl, nitro, cyano, aldehyde, acetyl, ethoxycarbonyl, difluoromethyl, trifluoromethyl;

[0018] (2) C1-C12 alkyl, C1-C12 alkoxy, C1-C12 alkylthio, C1-C12 alkylamino, C1-C12 oxaalkyl, C1-C12 thiaalkyl, C1-C12 azaalkyl, C1-C12 alkoxyacyl, C1-C12 alkylthioacyl, C1-C12 alkylazineacyl, C1-C12 acyloxy, C1-C12 acylthio or C1-C12 amide, wherein the above-mentioned alkyl refers to C1-C12 straight-chain or branched alkyl;

[0019] (3) Z2 and Z3 may also be substituted or unsubstituted five-membered rings or six-membered rings; the substituents may be selected from the same substituents as those for Z1;

[0020] (4) Z2 and Z3 may also be a five-membered ring or a six-membered ring substituted or unsubstituted with heteroatoms such as oxygen, sulfur, and nitrogen; the substituents may be selected from the same substituents as those for Z1;

[0021] 3) Wherein Z2, Z3, Z4, and Z5 are the same as those defined in 2) above;

[0022] 4) Wherein Z1, Z3, Z4, and Z5 are the same as those defined in 2) above;

[0023] 5)

[0024] Wherein, Y is nitrogen, sulfur or oxygen;

[0025] A1, A2, A3, A4, A5, A6 are each independently selected from:

[0026] (1) Hydrogen, fluorine, chlorine, bromine, iodine, methyl, benzyl, nitro, cyano, aldehyde, acetyl, ethoxycarbonyl, difluoromethyl, trifluoromethyl;

[0027] (2) C1-C12 alkyl, C1-C12 alkoxy, C1-C12 alkylthio, C1-C12 alkylnitro, C1-C12 oxaalkyl, C1-C12 thiaalkyl, C1-C12 azaalkyl, C1-C12 alkoxyacyl, C1-C12 alkylthioacyl, C1-C12 alkylnitroacyl, C1-C12 acyloxy, C1-C12 acylthio or C1-C12 amide, wherein the above-mentioned alkyl refers to a C1-C12 straight-chain or branched alkyl;

[0028] (3) A2 and A3 or A4 and A5 may also be a substituted or unsubstituted six-membered ring; the substituents may be selected from the same substituents as those mentioned above for A1;

[0029] 6) Wherein, Y, A7, A8 are the same as those defined in 5) above;

[0030] 7) R 2 Same as the definitions in 1)-6) above;

[0031] 8) R 3 Same as the definitions in 1)-6) above;

[0032] Preferably, the propiolate compound is a compound represented by the following structural formula:

[0033]

[0034] Preferably, the propiolate compound also includes its stereoisomer, its prodrug, or its pharmaceutically acceptable salt or pharmaceutically acceptable solvate, and a pharmaceutically acceptable carrier, diluent or excipient.

[0035] According to a specific technical solution of the present invention, the compound is a pharmaceutically acceptable salt, wherein the pharmaceutically acceptable salt is an inorganic salt or an organic salt, wherein the inorganic acid salt is a hydrochloride, a hydrobromide, a nitrate, a sulfate or a phosphate, and the organic acid salt is a formate, an acetate, a propionate, a benzoate, a maleate, a fumarate, a succinate, a tartrate, a citrate, an alkyl sulfonate or an aryl sulfonate.

[0036] On the other hand, the present invention also provides a pharmaceutical composition containing the above-mentioned propiolate compound, its stereoisomer, its prodrug, pharmaceutically acceptable salt, pharmaceutically acceptable solvate and a pharmaceutically acceptable carrier, diluent or excipient.

[0037] On the other hand, the present invention also provides a method for preparing the propiolate compound described in the above technical scheme, specifically, compound 1 and compound 2, or compound 3 and compound 2 are mixed and reacted to obtain a compound represented by general formula I or general formula II; the synthesis route is shown in (I) or (II) below:

[0038]

[0039] A can be an element such as carbon, oxygen, sulfur, selenium or tellurium, and R 1 ,R 2 ,R 3 Same as the definitions in 1)-8) above;

[0040] In the preparation of the compound represented by general formula I, the molar ratio of compound 1 to compound 2 is 1:1 to 4, preferably 1:2.

[0041] In the preparation of the compound represented by general formula II, the molar ratio of compound 3 to compound 2 is 1:1 to 4, preferably 1:2.

[0042] In the synthesis methods shown in the reaction schemes (I) and (II) of the present invention, the reaction conditions may be any of the following:

[0043] (1) Catalyst: dicyclohexylcarbodiimide, 4-dimethylaminopyridine, temperature: room temperature;

[0044] (2) Catalyst: 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 4-dimethylaminopyridine, temperature: room temperature;

[0045] (3) Catalyst: p-toluenesulfonic acid, temperature: 80-120°C;

[0046] The solvent used in the reaction conditions (1) to (3) is at least one of dichloromethane, toluene, chloroform, acetonitrile and the like.

[0047] The starting materials for this reaction can be purchased commercially or synthesized.

[0048] On the other hand, the synthesis of raw materials involved in the above synthesis scheme comprises the following steps: (1)

[0050]

[0051] Where A is oxygen or sulfur, R 1 Same as the definitions in 1)-6) above, reaction conditions: catalyst: potassium carbonate, temperature: 100-140°C; (2)

[0053]

[0054] Where A is sulfur, R 1 ,R 2 ,R 3 Same as the definitions in 1)-8) above, reaction conditions: catalyst: triethylamine, temperature: 0-4°C; (3)

[0056]

[0057] Where A is sulfur, R 1 ,R 2 ,R 3 Same as the definitions in 1)-8) above, reaction conditions: catalyst: triethylamine, temperature: 0-4°C; (4)

[0059]

[0060] Where A is oxygen, sulfur or tellurium, R 1 Same as the definitions in 1)-6) above, reaction conditions: catalyst: sodium borohydride, temperature: 60-90°C; (5)

[0062]

[0063] Where A is sulfur, selenium or tellurium, R 1 Same as the definitions in 1)-6) above, reaction conditions: (a) catalyst: copper oxide, potassium hydroxide, solvent: thionyl chloride, temperature: 90°C, (b) catalyst: sodium borohydride, temperature: 60-90°C; (6)

[0065]

[0066] Reaction conditions: Catalyst: sodium ethoxide, temperature: 80-120°C; (7)

[0068]

[0069] Reaction conditions: Temperature: 50-70°C.

[0070] The solvent used in steps (1) to (7) is at least one of N,N-dimethylformamide, ethanol and dichloromethane.

[0071] On the other hand, the present invention also provides a use of the above-mentioned propiolate compound, its stereoisomer, its prodrug, or pharmaceutically acceptable salt or pharmaceutically acceptable solvate in the preparation of a drug for preventing and treating bacteria or fungi related to agricultural and food pollution, especially in the preparation of a drug for preventing or treating methicillin-resistant Staphylococcus aureus. Preferably, the bacteria or fungi related to agricultural and food pollution are methicillin-resistant Staphylococcus aureus, Staphylococcus aureus, Listeria monocytogenes, Salmonella paratyphi B, Bacillus subtilis, Shigella flexneri, Vibrio parahaemolyticus, Penicillium digitatum, Aspergillus niger, etc.

[0072] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0073] The compounds provided by the present invention have different degrees of inhibitory effects on bacteria or fungi related to agricultural and food pollution, and especially have better antibacterial activity against methicillin-resistant Staphylococcus aureus. This also provides lead compounds for the development of drugs for preventing and treating bacteria or fungi related to agricultural and food pollution, especially drugs for preventing or treating methicillin-resistant Staphylococcus aureus, and has good development prospects. DETAILED DESCRIPTION

[0074] The embodiments of the present invention are described in detail below through specific examples, but they are not to be construed as limitations of the present invention in any way.

[0075]

[0076] The compounds of the above general formula are prepared synthetically in several categories.

[0077] General formula of compounds I and II

[0078]

[0079] Example 1

[0080] General synthetic formula of compound IA

[0081]

[0082] The starting materials for this reaction are commercially available.

[0083] Preparation method 1:

[0084] Compound 1 (0.2mmol), compound 2 (0.4mmol, 25μL), dicyclohexylcarbodiimide (0.24mmol, 49.5mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product IA was obtained by vacuum drying.

[0085] Preparation method 2:

[0086] Compound 1 (0.2mmol), compound 2 (0.4mmol, 25μL), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.24mmol, 46mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product IA was obtained by vacuum drying.

[0087] Preparation method 3:

[0088] Compound 1 (0.2 mmol), compound 2 (0.4 mmol, 25 μL), p-toluenesulfonic acid (0.3 mmol, 51.7 mg) and toluene (1 mL) were added to a 10 mL reaction tube. The reaction system was stirred at 100 ° C for 12 hours. After the reaction, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, and detected by TLC. The effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product IA was obtained by vacuum drying.

[0089] Compounds IB and IC can be synthesized using similar methods.

[0090] The table below lists the specific compounds and their structural identification data.

[0091]

[0092] Example 2

[0093] Synthetic formula of compound IC

[0094]

[0095] Preparation of compound 3

[0096]

[0097] Preparation method

[0098] Compound 1 (0.2mmol), compound 2 (0.8mmol, 40μL), sodium borohydride (0.3mmol, 11.3mg) and anhydrous ethanol (1mL) were added to a 10mL reaction tube. The reaction system was stirred at 80°C for 12 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product 3 was obtained by vacuum drying.

[0099] Preparation of Compound IC

[0100]

[0101] Preparation method 1:

[0102] Compound 3 (0.2mmol), compound 4 (0.4mmol, 25μL), dicyclohexylcarbodiimide (0.24mmol, 49.5mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, and it was extracted with water. The organic phase was dried and filtered, and the filtrate was evaporated to remove the solvent. The residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, and detected by TLC. The effluent containing the product was combined, and the solvent was distilled off by a rotary evaporator, and the target product IC was obtained by vacuum drying.

[0103] Preparation method 2:

[0104] Compound 3 (0.2mmol), compound 4 (0.4mmol, 25μL), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.24mmol, 46mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product IC was obtained by vacuum drying.

[0105] Preparation method 3:

[0106] Compound 3 (0.2 mmol), compound 4 (0.4 mmol, 25 μL), p-toluenesulfonic acid (0.3 mmol, 51.7 mg) and toluene (1 mL) were added to a 10 mL reaction tube. The reaction system was stirred at 100 ° C for 12 hours. After the reaction, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, and detected by TLC. The effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product IC was obtained by vacuum drying.

[0107] Compound ID can be synthesized using a similar method.

[0108] The table below lists the specific compounds and their structural identification data.

[0109]

[0110]

[0111] Example 3

[0112] General synthetic formula of compound IB-13

[0113] Preparation of compound 3

[0114]

[0115] Compound 1 (0.2 mmol), compound 2 (1 mL) and sodium ethoxide (0.3 mmol, 20.4 mg) were added to a 10 mL reaction tube. The reaction system was stirred at 100 ° C for 12 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, and it was extracted with water. The organic phase was dried and filtered, and the filtrate was evaporated to remove the solvent. The residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, and detected by TLC. The effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product 3 was obtained by vacuum drying.

[0116] Preparation of Compound IB-13

[0117]

[0118] Preparation method 1:

[0119] Compound 3 (0.2mmol), compound 4 (0.4mmol, 25μL), dicyclohexylcarbodiimide (0.24mmol, 49.5mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, and it was extracted with water. The organic phase was dried and filtered, and the filtrate was evaporated to remove the solvent. The residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, and detected by TLC. The effluent containing the product was combined, and the solvent was removed by rotary evaporation, and the target product IB-13 was obtained by vacuum drying.

[0120] Preparation method 2:

[0121] Compound 3 (0.2mmol), compound 4 (0.4mmol, 25μL), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.24mmol, 46mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product IB-13 was obtained by vacuum drying.

[0122] Preparation method 3:

[0123] Compound 3 (0.2mmol), compound 4 (0.4mmol, 25μL), p-toluenesulfonic acid (0.3mmol, 51.7mg) and toluene (1mL) were added to a 10mL reaction tube. The reaction system was stirred at 100°C for 12 hours. After the reaction, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product IB-13 was obtained by vacuum drying.

[0124] Compound IC-12 can be synthesized using a similar method.

[0125] The table below lists the specific compounds and their structural identification data.

[0126]

[0127] Example 4

[0128] Synthesis formula of compound IC-16

[0129]

[0130] Preparation of compound 4

[0131]

[0132] Compound 1 (0.2mmol, 12μL), compound 2 (0.2mmol, 17.6mg), compound 3 (0.4mmol, 20μL) and anhydrous ethanol (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 10min and then placed in a 60℃ oil bath for 24 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, and the mixture was extracted with water. The organic phase was dried and filtered, and the filtrate was evaporated to remove the solvent. The residue was chromatographed on a silica gel column and eluted with a mixed solvent of petroleum ether and ethyl acetate. The residue was detected by TLC. The effluent containing the product was combined, the solvent was removed by rotary evaporation, and the target product 4 was obtained by vacuum drying.

[0133] Preparation of compound IC-16

[0134]

[0135] Preparation method 1:

[0136] Compound 4 (0.2mmol), compound 5 (0.4mmol, 25μL), dicyclohexylcarbodiimide (0.24mmol, 49.5mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10ml reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, and it was extracted with water. The organic phase was dried and filtered, and the filtrate was evaporated to remove the solvent. The residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, and detected by TLC. The effluent containing the product was combined, and the solvent was removed by rotary evaporation, and the target product IC-16 was obtained by vacuum drying.

[0137] Preparation method 2:

[0138] Compound 4 (0.2mmol), compound 5 (0.4mmol, 25μL), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.24mmol, 46mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product IC-16 was obtained by vacuum drying.

[0139] Preparation method 3:

[0140] Compound 4 (0.2 mmol), compound 5 (0.4 mmol, 25 μL), p-toluenesulfonic acid (0.3 mmol, 51.7 mg) and toluene (1 mL) were added to a 10 mL reaction tube. The reaction system was stirred at 100 ° C for 12 hours. After the reaction, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, and detected by TLC. The effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product IC-16 was obtained by vacuum drying.

[0141] The table below lists the specific compounds and their structural identification data.

[0142]

[0143] Example 5

[0144] General synthetic formula of compound IB

[0145]

[0146] Preparation of compound 3

[0147]

[0148] Compound 1 (0.2mmol), compound 2 (0.4mmol, 35.2.6mg), potassium carbonate (0.3mmol, 41.5mg) and N,N-dimethylformamide (1mL) were added to a 10mL reaction tube. The reaction system was reacted at 120°C for 12 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, and it was extracted with water. The organic phase was dried and filtered, and the filtrate was evaporated to remove the solvent. The residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, and detected by TLC. The effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product 3 was obtained by vacuum drying.

[0149] Preparation of Compound IB

[0150]

[0151] Preparation method 1:

[0152] Compound 3 (0.2mmol), compound 4 (0.4mmol, 25μL), dicyclohexylcarbodiimide (0.24mmol, 49.5mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10ml reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, and it was extracted with water. The organic phase was dried and filtered, and the filtrate was evaporated by rotary evaporation to remove the solvent. The residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, and detected by TLC. The effluent containing the product was combined, and the solvent was removed by rotary evaporation, and the target product IB was obtained by vacuum drying.

[0153] Preparation method 2:

[0154] Compound 3 (0.2mmol), compound 4 (0.4mmol, 25μL), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.24mmol, 46mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product IB was obtained by vacuum drying.

[0155] Preparation method 3:

[0156] Compound 3 (0.2 mmol), compound 4 (0.4 mmol, 25 μL), p-toluenesulfonic acid (0.3 mmol, 51.7 mg) and toluene (1 mL) were added to a 10 mL reaction tube. The reaction system was stirred at 100°C for 12 hours. After the reaction was completed, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product IB was obtained by vacuum drying.

[0157] Compound IC can be synthesized using a similar method.

[0158] The table below lists the specific compounds and their structural identification data.

[0159]

[0160]

[0161]

[0162] Example 6

[0163] Synthetic formula of compound ID-3

[0164]

[0165] Preparation of compound 3

[0166]

[0167] Under nitrogen environment, compound 1 (0.2mmol, 35μL), compound 2 (0.4mmol, 31.6mg), copper oxide (0.02mmol, 1.6mg), potassium hydroxide (0.4mmol, 22.4mg) and dimethyl sulfoxide (1mL) were added to a 10mL reaction tube. The reaction system was reacted at 90°C for 6 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product 3 was obtained by vacuum drying.

[0168] Preparation of compound 5

[0169]

[0170] Compound 3 (0.2mmol), compound 4 (0.8mmol, 40μL), sodium borohydride (0.3mmol, 11.3mg) and anhydrous ethanol (1mL) were added to a 10ml reaction tube. The reaction system was stirred at 80°C for 12 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product 5 was obtained by vacuum drying.

[0171] Preparation of compound ID-3

[0172]

[0173] Preparation method 1:

[0174] Compound 5 (0.2mmol), compound 6 (0.4mmol, 25μL), dicyclohexylcarbodiimide (0.24mmol, 49.5mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product ID-3 was obtained by vacuum drying.

[0175] Preparation method 2:

[0176] Compound 5 (0.2mmol), compound 6 (0.4mmol, 25μL), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.24mmol, 46mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product ID-3 was obtained by vacuum drying.

[0177] Preparation method 3:

[0178] Compound 5 (0.2mmol), compound 6 (0.4mmol, 25μL), p-toluenesulfonic acid (0.3mmol, 51.7mg) and toluene (1mL) were added to a 10mL reaction tube. The reaction system was stirred at 100°C for 12 hours. After the reaction, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product ID-3 was obtained by vacuum drying.

[0179] Compound IE-4 can be synthesized using a similar method.

[0180] The table below lists the specific compounds and their structural identification data.

[0181]

[0182] Example 7

[0183] General synthetic formula of compound IF

[0184]

[0185] Preparation of compound 3

[0186]

[0187] Compound 1 (0.2mmol, 34ul), compound 2 (0.4mmol), triethylamine (0.4mmol, 56μL) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at 0°C for 3 hours. After the reaction was completed, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product 3 was obtained by vacuum drying.

[0188] Preparation of Compound IF

[0189]

[0190] Preparation method 1:

[0191] Compound 3 (0.2mmol), compound 4 (0.4mmol, 25μL), dicyclohexylcarbodiimide (0.24mmol, 49.5mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporation, and the target product IF was obtained by vacuum drying.

[0192] Preparation method 2:

[0193] Compound 3 (0.2mmol), compound 4 (0.4mmol, 25μL), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.24mmol, 46mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product IF was obtained by vacuum drying.

[0194] Preparation method 3:

[0195] Compound 3 (0.2 mmol), compound 4 (0.4 mmol, 25 μL), p-toluenesulfonic acid (0.3 mmol, 51.7 mg) and toluene (1 mL) were added to a 10 mL reaction tube. The reaction system was stirred at 100 ° C for 12 hours. After the reaction, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product IF was obtained by vacuum drying.

[0196] The table below lists the specific compounds and their structural identification data.

[0197]

[0198] Example 8

[0199] General Synthesis Formula of Compound IIA-1

[0200]

[0201] Preparation of compound 3

[0202]

[0203] Compound 1 (0.2mmol, 34ul), compound 2 (0.4mmol, 40μL), triethylamine (0.4mmol, 56μL) and dichloromethane (1mL) were added to a 10ml reaction tube. The reaction system was stirred at 0°C for 3 hours. After the reaction, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product 3 was obtained by vacuum drying.

[0204] Preparation of Compound IIA-1

[0205]

[0206] Preparation method 1:

[0207] Compound 3 (0.2mmol), compound 4 (0.4mmol, 25μL), dicyclohexylcarbodiimide (0.24mmol, 49.5mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, it was cooled, an appropriate amount of ethyl acetate was added, and it was extracted with water. The organic phase was dried and filtered, and the filtrate was evaporated to remove the solvent. The residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, and detected by TLC. The effluent containing the product was combined, and the solvent was distilled off by a rotary evaporator, and the target product IIA-1 was obtained by vacuum drying.

[0208] Preparation method 2:

[0209] Compound 3 (0.2mmol), compound 4 (0.4mmol, 25μL), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.24mmol, 46mg), 4-dimethylaminopyridine (0.02mmol, 2.4mg) and dichloromethane (1mL) were added to a 10mL reaction tube. The reaction system was stirred at room temperature for 12 hours. After the reaction was completed, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, the filtrate was rotary evaporated, the solvent was removed, the residue was chromatographed on a silica gel column, eluted with a mixed solvent of petroleum ether and ethyl acetate, detected by TLC, the effluent containing the product was combined, the solvent was removed by rotary evaporator, and the target product IIA-1 was obtained by vacuum drying.

[0210] Preparation method 3:

[0211] Compound 3 (0.2 mmol), compound 4 (0.4 mmol, 25 μL), p-toluenesulfonic acid (0.3 mmol, 51.7 mg) and toluene (1 mL) were added to a 10 mL reaction tube. The reaction system was stirred at 100 ° C for 12 hours. After the reaction, an appropriate amount of ethyl acetate was added, extracted with water, the organic phase was dried and filtered, and the filtrate was evaporated to remove the solvent. The residue was chromatographed on a silica gel column and eluted with a mixed solvent of petroleum ether and ethyl acetate. TLC was detected, and the effluent containing the product was combined, the solvent was distilled off by a rotary evaporator, and the target product IIA-1 was obtained by vacuum drying.

[0212] The table below lists the specific compounds and their structural identification data.

[0213]

[0214] Biological activity test, selection of new propiolate compounds for antibacterial test

[0215] 1. Experimental Materials

[0216] Bacterial strains: Methicillin-resistant Staphylococcus aureus ATCC43300, Staphylococcus aureus ATCC6538, Listeria monocytogenes ATCC19115, Salmonella paratyphi CMCC(B)50094, Bacillus subtilis ATCC6633, Shigella Castellani CMCC(B)51572, Vibrio Parahemolyticus ATCC17802.

[0217] Fungal strains: Penicillium digitatum CGMCC3.5752, Aspergillus niger CMCC(F)98003.

[0218] Culture medium: LB broth medium (Guangdong Huankai Microbiological Technology Co., Ltd.), potato dextrose agar medium (Guangdong Huankai Microbiological Technology Co., Ltd.) and potato dextrose broth medium (Guangdong Huankai Microbiological Technology Co., Ltd.).

[0219] Positive controls: penicillin, vancomycin, carbendazim.

[0220] 2. Experimental Methods

[0221] Bacteria: Use tweezers to pick up a single colony on the plate with a sterilized toothpick, place it in a sterile liquid LB medium (4 mL test tube), and culture it in a shaking incubator at 37°C overnight to activate the bacteria; then transfer 80-500 μL of the bacterial solution to a new liquid culture medium (the transfer amount depends on the growth status of the bacteria), and culture it in a shaking incubator at 37°C for 2-6 hours. Dilute the bacterial solution to McFarland standard 0.5, and then dilute it 20 times with sterile water for later use.

[0222] Add the compound prepared at a concentration of 2 mg / mL to a 96-well plate, 2.5 μL per well. Use a spray gun to take the bacterial solution and add it to the 96-well plate. The final drug concentration is 100, 50, 25, 12.5, 6.25, 3.13, 1.56, and 0.78 μg / mL, with 3 parallel replicates. Incubate at 37°C in a shaker for 16 to 20 hours, and observe the lowest concentration that inhibits bacterial growth, the minimum inhibitory concentration (MIC).

[0223] Fungi: Pick fungal cells from the fungal culture slant and inoculate them onto potato dextrose agar plates by the spot grafting method. Place them in a 28°C incubator for several days to activate the fungus. Then collect spores, wash them out of the plates with sterile 0.9% saline, filter them with a cell sieve, and dilute to McFarland standard 0.5 for later use.

[0224] Take 50 μL of spores, add them to 50 mL of potato dextrose broth, mix well, add them to a 96-well plate, add 0.2 mL to each well; then add the pre-prepared compound with a concentration of 2 mg / ml to the 96-well plate, the final drug concentration is 100, 50, 25, 12.5, 6.25, 3.13, 1.56, 0.78 μg / mL, set up 3 parallel replicates. Incubate at 28°C in a shaker for 48 to 72 hours, and observe the lowest concentration that inhibits bacterial growth, the minimum inhibitory concentration (MIC), with the naked eye.

[0225] 3. Experimental results

[0226] Penicillin, vancomycin and carbendazim were used as positive controls to preliminarily explore the antibacterial effects of the tested drugs. The results are shown in the following table:

[0227]

[0228]

[0229] MA, methicillin-resistant Staphylococcus aureus; SA, Staphylococcus aureus; LM, Listeria monocytogenes; SP, Salmonella paratyphi; BS, Bacillus subtilis; SC, Shigella Castellani; VP, Vibrio parahemolyticus; PD, Penicillium digitatum; AN, Aspergillus niger.

[0230] The present invention screens the antibacterial activity of the above-selected compounds. The experimental results show that the selected compounds have different degrees of inhibitory effects on the tested strains, especially have better antibacterial activity against methicillin-resistant Staphylococcus aureus. This also provides lead compounds for the development of drugs for preventing and treating bacteria or fungi related to agricultural and food pollution, especially for the prevention or treatment of methicillin-resistant Staphylococcus aureus, and has good development prospects.

[0231] The above are only some embodiments of the present invention, but the embodiments of the present invention are not limited by the above embodiments. For those skilled in the art, several modifications and improvements can be made without departing from the creative concept of the present invention, which all belong to the protection scope of the present invention.

Claims

1. A propiolate compound, characterized in that The propiolate compound is a compound represented by the following structural formula:

2. A pharmaceutical composition, characterized in that It comprises the propiolate compound according to claim 1, its stereoisomer, its prodrug, pharmaceutically acceptable salt, pharmaceutically acceptable solvate and pharmaceutically acceptable carrier, diluent or excipient.

3. Use of the propiolate compound according to claim 1 or the pharmaceutical composition according to claim 2 in the preparation of a drug for preventing and treating bacteria or fungi associated with agricultural and food pollution, wherein the bacteria or fungi associated with agricultural and food pollution are methicillin-resistant Staphylococcus aureus, Staphylococcus aureus, Listeria monocytogenes, Salmonella paratyphi B, Bacillus subtilis, Shigella flexneri, Vibrio parahaemolyticus or Penicillium digitatum.

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