Coumarin compounds, methods of making and using the same

Abstract

The application discloses coumarin compounds, and a preparation method and application thereof. The coumarin compound is selected from a compound shown in formula (A-A); wherein R1 is selected from a group shown in the formula, H or C1-C10 alkyl, wherein R 1′ oxalyl chloride ester selected from H or C1-C10, R 1” substituted or unsubstituted aryl; R2 is selected from a group shown in the formula or H, wherein R 2′ substituted or unsubstituted aryl; R3 is selected from a group shown in the formula, H or hydroxyl, wherein R 3′ C1-C10 carboxylic acid, R 3” substituted or unsubstituted aryl; and when R1 of the coumarin compound is selected from hydrogen, R3 cannot be selected from hydroxyl. The coumarin compound has simple structure, small molecular weight and good biological activity, can inhibit test bacteria in different degrees, and can be used as a potential new antibacterial agent.

Description

Technical Field

[0001] This invention relates to the technical field of plant antibacterial agents, and more specifically, to coumarin compounds, their preparation methods, and applications. Background Technology

[0002] Using natural active products as lead compounds and modifying their structures to obtain substances with superior properties has become a research hotspot worldwide. Coumarin, scientifically known as benzo-α-pyranone, is an oxygen-containing heterocyclic compound. Coumarin and its derivatives are widely distributed in nature, existing as secondary metabolites in the roots, flowers, leaves, bark, seeds, and fruits of many plants. These compounds possess advantages such as small molecular weight, relatively simple synthesis, broad pharmacological effects, and low toxicity, attracting widespread attention from scholars both domestically and internationally.

[0003] Many reported coumarin-based natural and synthetic compounds possess a wide range of pharmacological activities, such as antibacterial, antioxidant, anticancer, antituberculosis, antiproliferative, and enzyme-inhibiting activities. Currently, drugs containing effective coumarin derivatives are available on the market, such as leucin (antibiotic), warfarin (anticoagulant), phenylpropanol (anticoagulant), cyclosporine (anticholinergic and anticonvulsant), and methoxsalen (anti-psoriasis).

[0004] Plant diseases are a global threat to crop growth and the ecological environment. Crops are susceptible to many fungal infections, leading to reduced yields and significant economic losses. Therefore, developing new antifungal drugs is one way to address this problem and improve crop productivity.

[0005] Therefore, it is necessary to develop a new compound with better antibacterial effects. Summary of the Invention

[0006] To address the problems existing in the prior art, this invention proposes coumarin compounds, their preparation methods, and applications. The coumarin compounds of this invention have simple structures and small molecular weights; they possess good biological activity and exhibit varying degrees of inhibition against the tested bacteria, making them potential novel antibacterial agents.

[0007] One object of the present invention is to provide a coumarin compound selected from compounds represented by formula (AA);

[0008]

[0009] in,

[0010] R1 is selected from The indicated group, H or C1-C 10 Alkyl groups;

[0011] The R 1′ Selected from H, Among them, R 1-1′ Selected from alkyl and alkenyl groups; R 1-2′ Selected from alkenyl groups;

[0012] The R 1″ Selected from substituted or unsubstituted aryl groups;

[0013] R2 is selected from The group or H shown, where R 2′ Selected from substituted or unsubstituted aryl groups;

[0014] The R3 is selected from H, hydroxyl, ester group or... The group shown, wherein R 3′ The aryl group is selected from substituted or unsubstituted aryl groups; and when R1 of the coumarin compound is selected from hydrogen, R3 cannot be selected from hydroxyl groups.

[0015] In the coumarin compounds described in this invention, preferably,

[0016] R1 is selected from The indicated group, H, or C1-C5 alkyl group; the R 1′ Selected from H, Among them, R 1-1′ Selected from C1-C6 alkyl and C1-C6 alkenyl groups; R 1-2′ Selected from C1-C6 alkenyl groups; and / or,

[0017] R 1″ Selected from C6-C 10 Substituted or unsubstituted phenyl groups; and / or,

[0018] R2 is selected from The group or H shown, where R 2′ Selected from C5-C 10 Substituted or unsubstituted aryl groups; and / or,

[0019] R3 is selected from H, hydroxyl, C2-C. 10 ester group or The group shown, wherein R 3′ Selected from C4-C 10 The aryl group may be substituted or unsubstituted; and when R1 of the coumarin compound is selected from hydrogen, R3 cannot be selected from hydroxyl.

[0020] In the coumarin compounds described in this invention, preferably,

[0021] R1 is selected from The indicated group, H or C1-C3 alkyl group, said R1′ Selected from H, Among them, R 1-1′ Selected from C1-C3 alkyl groups and C2-C4 alkenyl groups; R 1-2′ Selected from C2-C4 alkenyl groups; and / or,

[0022] R 1″ A phenyl group selected from at least one substituent of halogens, hydroxyl groups, and C1-C5 alkoxy groups; and / or,

[0023] R2 is selected from The group shown, wherein R 2′ An aryl group selected from at least one substituent chosen from halogens, hydroxyl groups, C1-C5 fluoroalkyl groups, and C1-C5 alkoxy groups; and / or,

[0024] The R3 is selected from H, hydroxyl, C2-C8 ester groups, or... The group shown, wherein R 3′ The aryl group is substituted with at least one of the following: halogen, hydroxyl, or C1-C5 fluoroalkyl; and when R1 of the coumarin compound is selected from hydrogen, R3 cannot be selected from hydroxyl.

[0025] In the coumarin compounds described in this invention, preferably,

[0026] R1 is selected from The indicated group, H or C1-C2 alkyl group, said R 1′ Selected from H, Among them, R 1-1′ Selected from C1-C2 alkyl groups and C3-C4 alkenyl groups; R 1-2′ Selected from C3-C4 alkenyl groups; and / or, R 1″ A phenyl group selected from at least one substituent of halogen, hydroxyl, and C1-C2 alkoxy groups, preferably R. 1″ The number of substituents is ≥2, and the substituents are at least ortho- and para-substituents; and / or,

[0027] R2 is selected from The group shown, wherein R 2′ An aryl group substituted with at least one substituent selected from halogens, hydroxyl groups, C1-C2 fluoroalkyl groups, and C1-C2 alkoxy groups, wherein the aryl group is phenyl, pyridyl, or pyrroleyl; preferably R 2′ The number of substituents is ≥2, and the substituents are at least ortho- and para-substituents; and / or,

[0028] The R3 is selected from H, hydroxyl, C2-C6 ester group or... The group shown, wherein R 3′The aryl group is substituted with at least one of the following: halogen, hydroxyl, or C1-C2 fluoroalkyl, wherein the aryl group is phenyl, pyridyl, or pyrrolithyl; preferably, the substituent is ortho-substituted and / or para-substituted; and when R1 of the coumarin compound is selected from hydrogen, R3 cannot be selected from hydroxyl.

[0029] In the coumarin compounds described in this invention, preferably,

[0030] In the coumarin compounds, R1 or R2 is selected from hydrogen; and / or,

[0031] The coumarin compounds are selected from the compounds shown in formula (I) and formula (II);

[0032]

[0033] Wherein, R1, R2, and R3 correspond to the same R1, R2, and R3 of the coumarin compounds described in any one of the objectives of this invention.

[0034] In the coumarin compounds described in this invention, preferably:

[0035]

[0036]

[0037]

[0038]

[0039] A second objective of this invention is to provide a method for preparing coumarin compounds as described in any one of the objectives of this invention, wherein the compound is selected from at least one of the following methods:

[0040] Method (1):

[0041] Optionally, in step (1-1), the compound shown in formula A, R3-H, a carboxylic acid activator, and a catalyst are mixed in a solvent and reacted at room temperature to generate the compound shown in formula (A-1); wherein the structure of the compound shown in formula A is as follows:

[0042] R2 corresponds to the same R2 in any of the coumarin compounds described in any one of the objectives of this invention; preferably, R2 is selected from H;

[0043] The structure of the compound represented by formula (A-1) is shown below:

[0044] R2 corresponds to the same R2 in any of the coumarin compounds described in any one of the objectives of this invention; preferably, R2 is selected from H;

[0045] R3 is selected from ester groups;

[0046] Step (1-2) After mixing the compound shown in formula A or the compound shown in formula (A-1) and the weak organic base in a solvent, add R dropwise. 1′ -X heating reaction yields product 1, which is a coumarin compound; wherein, R 1′ R in -X 1′ R in any of the coumarin compounds according to any one of the objectives of this invention 1′ The corresponding ones are the same;

[0047] Optionally, steps (1-3) include: further reacting product 1, methoxyamine hydrochloride, and an organic weak base in a solvent to obtain the coumarin compound;

[0048] Method (2):

[0049] Or, the compound shown in formula A, R 1″ =O, an organic weak acid, reacted in a solvent to obtain the coumarin compounds described above; R 1″ =R in O 1″ R in any of the coumarin compounds according to any one of the objectives of this invention 1″ The corresponding ones are the same;

[0050] Method (3):

[0051] Or, the compound shown in formula B, R 3′ =O, an organic weak acid, reacted in a solvent to obtain the coumarin compounds described above;

[0052] The structure of the compound represented by formula B is shown below:

[0053]

[0054] R1 is selected from C1-C 10 Alkyl group; preferably R1 is selected from methyl;

[0055] R2 corresponds to the same R2 in any of the coumarin compounds described in any one of the objectives of this invention; preferably, R2 is selected from H;

[0056] R 3′ =R in O 3′ R in any of the coumarin compounds according to any one of the objectives of this invention 3′ The corresponding ones are the same;

[0057] Method (4):

[0058] The compound shown in formula C, R 2′=O, an organic weak acid reacts with heat in a solvent to produce the aforementioned coumarin compounds;

[0059] The structure of the compound shown in formula C is as follows:

[0060] R1 corresponds to the same R1 in any of the coumarin compounds described in any one of the objectives of this invention; R1 is preferably selected from H;

[0061] R3 corresponds to the same R3 in any of the coumarin compounds described in any one of the objectives of this invention; R3 is selected from H; R3 is preferably selected from H.

[0062] R 2′ =R in O 2′ R in any of the coumarin compounds according to any one of the objectives of this invention 2′ The correspondence is the same.

[0063] In the preparation method of coumarin compounds described in this invention, preferably,

[0064] The solvents in methods (1), (2), (3), and (4) are each independently selected from at least one of methanol, ethanol, or dichloromethane; and / or,

[0065] The organic weak base in method (1) is selected from at least one of pyridine or triethylamine; and / or,

[0066] The organic weak acid in methods (2), (3), and (4) is selected from glacial acetic acid; and / or,

[0067] In method (1), the catalyst is selected from at least one of DMAP (4-dimethylaminopyridine) and DCC (dicyclohexylcarbodiimide); and / or,

[0068] In method (1), the carboxylic acid activator is selected from 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide; and / or,

[0069] In method (1), the reaction temperature in steps (1-1) and (1-2) is room temperature; the reaction time is independently selected from 4 to 10 hours; the mixing in step (1-2) is carried out in an ice-water bath; and / or,

[0070] In method (1), the reaction temperature in steps (1-3) is 0-30℃; the reaction time is 30 min-8 h; and / or,

[0071] In method (1), in step (1-1),

[0072] The molar ratio of the compound shown in Formula A to R3-H is 1:(1-3); preferably 1:(1.5-2.5); and / or,

[0073] The molar ratio of the compound shown in Formula A to the catalyst is 1:(0.15–5); preferably 1:(0.15–3); and / or,

[0074] The molar ratio of the compound shown in Formula A to the carboxylic acid activator is 1:(1-3); preferably 1:(1.5-2.5); and / or,

[0075] The molar volume ratio of the compound shown in Formula A to the solvent is 1 mmol : (1-10) mL; and / or,

[0076] In method (1), in steps (1-2),

[0077] The compound shown in formula A or the compound shown in formula (A-1) and R 1′ The molar ratio of -X is 1:(1-5); preferably 1:(1-2); and / or,

[0078] The molar ratio of the compound shown in formula A or the compound shown in formula (A-1) to the weak organic base is 1:(0.02-2); preferably 1:(0.02-1); and / or,

[0079] The molar volume ratio of the compound shown in Formula A or the compound shown in Formula (A-1) to the solvent is 1 mmol: (1-10) mL; and / or,

[0080] In method (1), in steps (1-3),

[0081] The molar ratio of the compound shown in product 1 to methoxyamine hydrochloride is 1:(1-5); preferably 1:(1-2); and / or,

[0082] The molar ratio of the compound shown in product 1 to the organic weak base is 1:(0.02–2); preferably 1:(0.02–1); and / or,

[0083] The molar volume ratio of the compound shown in product 1 to the solvent is 1 mmol: (1-10) mL.

[0084] In the preparation method of coumarin compounds described in this invention, preferably,

[0085] In method (2),

[0086] The compound shown in Formula A and R 1″ =O molar ratio is 1:(1-5); preferably 1:(1-2); and / or,

[0087] The molar volume ratio of the compound shown in Formula A to the weak organic acid is 1 mmol : (0.02–4) mL; preferably 1 mmol : (2–3) mL; and / or,

[0088] The molar volume ratio of the compound shown in Formula A to the solvent is 1 mmol : (1-20) mL; and / or,

[0089] The reaction temperature is 50–90°C; preferably 50–80°C; and / or,

[0090] The reaction time is 4–10 h; preferably 4–8 h; and / or,

[0091] In method (3),

[0092] The compound shown in formula B and R 3′ =O molar ratio is 1:(1-5); preferably 1:(1-2); and / or,

[0093] The molar ratio of the compound shown in Formula B to the weak organic acid is 1 mmol : (0.02–4) mL; preferably 1 mmol : (2–3) mL; and / or,

[0094] The molar volume ratio of the compound shown in Formula B to the solvent is 1 mmol : (1-20) mL; and / or,

[0095] The reaction temperature is 50–90°C; preferably 50–80°C; and / or,

[0096] The reaction time is 4–10 h; preferably 4–8 h; and / or,

[0097] In method (4),

[0098] The compound shown in formula C and R 2′ =O molar ratio is (0.7-1):1; preferably (0.75-0.9):1; and / or,

[0099] The molar volume ratio of the compound shown in Formula C to the weak organic acid is 1 mmol:(1–3) mL; preferably 1 mmol:(1.5–2.5) mL; and / or,

[0100] The molar volume ratio of the compound shown in Formula C to the solvent is 1 mmol:(4–20) mL; preferably 1 mmol:(8–15) mL; and / or,

[0101] The reaction temperature is 60-100℃; preferably 50-80℃; and / or,

[0102] The reaction time is 5-10 hours; preferably 4-8 hours.

[0103] This invention involves the condensation reaction of primary amine compounds with simple aldehydes and ketones to obtain molecules containing imine groups in their molecular structure. One class of compounds is called Schiff bases, which undergo amide condensation reactions with acyl chlorides to form compounds containing amide bonds in their molecular structure. A class of compounds that undergo esterification with carboxylic acids to form molecules containing ester bonds. A class of compounds. Due to the presence of imine in the Schiff base structure, Schiff bases possess high biological activity, including antiviral and antibacterial activities; amide and ester bonds are also effective antibacterial active groups. Based on this, in combination with other substituent groups on the coumarin skeleton, a series of compounds with potential biological activity and even better performance can be obtained.

[0104] A third objective of this invention is to provide the application of the coumarin compounds described in the first objective of this invention or the coumarin compounds prepared by the preparation method described in the second objective of this invention in antibacterial agents; preferably in antibacterial agents against at least one pathogen selected from Botrytis cinerea, Alternaria solanacearum, Fusarium oxysporum, and Alternaria; more preferably in antibacterial agents against plants.

[0105] The endpoints and any values ​​of the ranges disclosed in this invention are not limited to the precise ranges or values; these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein. In the following, various technical solutions can, in principle, be combined with each other to obtain new technical solutions, which should also be considered as specifically disclosed herein.

[0106] Compared with the prior art, the present invention has at least the following advantages:

[0107] (1) This invention establishes a method for synthesizing coumarin compounds and identifies their structures. The coumarin compounds of this invention have simple structures and small molecular weights; they have good biological activity and inhibit the test bacteria to varying degrees, and can be used as potential novel antibacterial agents. To date, no coumarin compounds identical to those of this invention and their antibacterial applications against plant-derived fungi have been published.

[0108] (2) The synthesis method of the present invention is simple and has few steps, and has high separation. Detailed Implementation

[0109] The present invention will now be described in detail with reference to specific embodiments. It should be noted that the following embodiments are only used to further illustrate the present invention and should not be construed as limiting the scope of protection of the present invention. Some non-essential improvements and adjustments made by those skilled in the art based on the content of the present invention are still within the scope of protection of the present invention.

[0110] It should also be noted that the various specific technical features described in the following embodiments can be combined in any suitable manner without contradiction. To avoid unnecessary repetition, the various possible combinations will not be described separately in this invention.

[0111] Furthermore, various embodiments of the present invention can be combined in any way, as long as they do not violate the spirit of the present invention. The resulting technical solutions are part of the original disclosure of this specification and also fall within the protection scope of the present invention.

[0112] Unless otherwise specified, the raw materials used in the examples and comparative examples are all disclosed in the prior art, such as those that can be directly purchased or prepared according to the preparation methods disclosed in the prior art.

[0113] Example 1

[0114] 1. Synthesis of 7-hydroxy-4-aminocoumarin (the structure of this compound is known).

[0115]

[0116] At room temperature, resorcinol (22.84 g, 0.2 mol), cyanoacetic acid (19.33 g, 0.2 mol), zinc chloride (13.95 g, 0.1 mmol), and diethyl ether (50 mL) were added sequentially to a dry three-necked round-bottom flask. A condenser and a drying tube containing anhydrous calcium chloride were attached. Hydrogen chloride gas and nitrogen gas were continuously introduced, and the reaction was stirred vigorously for 5 hours. After that, a large amount of white solid precipitate appeared in the three-necked round-bottom flask. The reaction was stopped, quenched with distilled water, and the white solid was filtered and dried. The yield of the target product was 23.7% (8.41 g), and the melting range was 272.2-273 °C.

[0117] 2. Synthesis of compound 1:

[0118]

[0119] 4-Amino-7-hydroxycoumarin (100 mg, 0.56 mmol), triethylamine (131.5 mg, 1.3 mmol), and ultra-dry dichloromethane (5 mL) were added to a dry round-bottom flask at room temperature. The flask was cooled to 5 °C in an ice-water bath and reacted for 30 min. Then, oxaloyl chloride monoallyl ester (mg, mmol) was slowly added dropwise. After the addition was complete, the temperature was slowly raised to room temperature and reacted for 8 h. After the reaction was completed, the mixture was quenched with distilled water and then extracted three times with dichloromethane. The organic phases were combined. Compound 1 was purified by column chromatography using dichloromethane:methanol = 50:1 as the eluent. The yield of the target product was 64.6%, and the melting range was 231-234 °C.

[0120] Example 2

[0121] Synthesized compound 2:

[0122]

[0123] Compound 1 (223.68 mg, 0.75 mmol), methoxyamine hydrochloride (75 mg, 0.9 mmol), and pyridine (79 mg, 1 mmol) were sequentially added to a 25 mL round-bottom flask equipped with a stirrer and a condenser under an oil bath at 80 °C. Methanol was used as the solvent (5 mL), and the reaction was stopped after 3 hours. After standing for half an hour, recrystallization yielded compound 2. The yield of the target product was 43.5%, and the melting range was 198-203 °C.

[0124] Example 3

[0125] Synthesized compound 3:

[0126]

[0127] At room temperature, 105.2 mg (1.0300 mmol) of 2-methylbutyric acid, 156.1 mg (1.0055 mmol) of EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide), and 18.8 mg (0.1539 mmol) of DMAP were added sequentially to a 50 mL round-bottom flask. Dichloromethane was used as the solvent (3 mL). After reacting for 20 min, 89.1 mg (0.5029 mmol) of 4-amino-7-hydroxycoumarin was weighed and added to the reaction flask. The reaction was continued for 8 h and then stopped. Compound 3 was obtained by column chromatography using dichloromethane:methanol = 50:1 as the eluent. The yield of the target product was 85.3%. The melting range was 208-210 °C.

[0128] Example 4

[0129] Synthesized compound 4:

[0130]

[0131] Compound 3 (200 mg, 0.7654 mmol), triethylamine (88.16 mg, 0.8712 mmol), and ultra-dry dichloromethane (5 mL) were added to a dry round-bottom flask at room temperature. The flask was cooled to 5 °C in an ice-water bath and reacted for 30 min. Then, methyl oxaloyl chloride (94.5 mg, 0.7747 mmol) was slowly added dropwise. After the addition was complete, the temperature was slowly raised to room temperature and reacted for 8 h. Compound 4 was purified by column chromatography using dichloromethane:ethyl acetate (30:1) as the eluent. The yield of the target product was 37.3%. The melting range was 169-171 °C.

[0132] Example 5

[0133] Synthesized compound 5:

[0134]

[0135] Compound 3 (200.1 mg, 0.7658 mmol), triethylamine (88.16 mg, 0.8712 mmol), and ultra-dry dichloromethane (5 mL) were added to a dry round-bottom flask at room temperature. The flask was cooled to 5 °C in an ice-water bath and reacted for 30 min. Then, ethyl oxaloyl chloride (135.3 mg, 0.9768 mmol) was slowly added dropwise. After the addition was complete, the temperature was slowly raised to room temperature and reacted for 8 h. Compound 5 was purified by column chromatography using dichloromethane:ethyl acetate (30:1) as the eluent. The yield of the target product was 57.3%. The melting range was 140-144 °C.

[0136] Example 6

[0137]

[0138] Compound 3 (200.1 mg, 0.7658 mmol), triethylamine (88.16 mg, 0.8712 mmol), and ultra-dry dichloromethane (5 mL) were added to a dry round-bottom flask at room temperature. The flask was cooled to 5 °C in an ice-water bath and reacted for 30 min. Then, oxaloyl chloride monoallyl ester (122.8 mg, 0.8267 mmol) was slowly added dropwise. After the addition was complete, the temperature was slowly raised to room temperature and reacted for 8 h. Compound 6 was purified by column chromatography using dichloromethane:ethyl acetate (30:1) as the eluent. The yield of the target product was 13.7%. The melting range was 151-156 °C.

[0139] Example 7

[0140]

[0141] At room temperature, 119.6 mg (1.03 mmol) of 2-methylvaleric acid, 156.0 mg (1.00 mmol) of EDC, and 18.8 mg (0.15 mmol) of DMAP were added sequentially to a 50 mL round-bottom flask. Using dichloromethane as solvent, (3) mL of solution was added. After reacting for 20 min, 89.0 mg (0.50 mmol) of 4-amino-7-hydroxycoumarin was weighed and added to the reaction flask. The reaction was continued for 8 h before stopping. Compound 7 was obtained by column chromatography using dichloromethane:methanol = 50:1 as the eluent. The yield of the target product was 84.4%, and the melting range was 188-191 °C.

[0142] Example 8

[0143]

[0144] Compound 7 (212 mg, 0.77 mmol), triethylamine (88.16 mg, 0.8712 mmol), and ultra-dry dichloromethane (5 mL) were added to a dry round-bottom flask at room temperature. The flask was cooled to 5 °C in an ice-water bath and reacted for 30 min. Then, methyl oxaloyl chloride (94.5 mg, 0.7747 mmol) was slowly added dropwise. After the addition was complete, the temperature was slowly raised to room temperature and reacted for 8 h. Compound 8 was purified by column chromatography using dichloromethane:ethyl acetate (30:1) as the eluent. The yield of the target product was 55.7%, and the melting range was 148-151 °C.

[0145] Example 9

[0146]

[0147] Compound 7 (212 mg, 0.77 mmol), triethylamine (88.16 mg, 0.8712 mmol), and ultra-dry dichloromethane (5 mL) were added to a dry round-bottom flask at room temperature. The flask was cooled to 5 °C in an ice-water bath and reacted for 30 min. Then, ethyl oxaloyl chloride (135.3 mg, 0.9768 mmol) was slowly added dropwise. After the addition was complete, the temperature was slowly raised to room temperature and reacted for 8 h. Compound 9 was purified by column chromatography using dichloromethane:ethyl acetate (30:1) as the eluent. The yield of the target product was 36.4%, and the melting range was 151-156 °C.

[0148] Example 10

[0149]

[0150] Compound 7 (212 mg, 0.77 mmol), triethylamine (88.16 mg, 0.8712 mmol), and ultra-dry dichloromethane (5 mL) were added to a dry round-bottom flask at room temperature. The flask was cooled to 5 °C in an ice-water bath and reacted for 30 min. Then, oxaloyl chloride monoallyl ester (122.8 mg, 0.8267 mmol) was slowly added dropwise. After the addition was complete, the temperature was slowly raised to room temperature and reacted for 8 h. Compound 10 was purified by column chromatography using dichloromethane:ethyl acetate (30:1) as the eluent. The yield of the target product was 27.4%, and the melting range was 158-163 °C.

[0151] Example 11

[0152]

[0153]

[0154] Under oil bath conditions at 80°C, 100 mg (0.56 mmol) of 4-amino-7-hydroxycoumarin, 146.2 mg (0.72 mmol) of 2-fluoro-4-bromobenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol was used as the solvent (10 mL). The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, and compound 11 was obtained by filtration. The yield of the target product was 25.16%, and the melting range was 293-296°C.

[0155] Example 12

[0156]

[0157] Under oil bath conditions at 80°C, 4-methyl-7-aminocoumarin (131 mg, 0.75 mmol), 4-hydroxy-3-pyridinecarboxaldehyde (123 mg, 1.0 mmol), and glacial acetic acid (1 mL) were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol (10 mL) was used as the solvent. The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, which was filtered to obtain compound 12. The yield of the target product was 10.5%.

[0158] Example 13

[0159]

[0160] Under oil bath conditions at 80°C, 131 mg (0.75 mmol) of 4-methyl-7-aminocoumarin, 95 mg (1.0 mmol) of 2-trifluoromethyl-3-pyridinecarboxaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol was used as the solvent. The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, which was filtered to obtain compound 13. The yield of the target product was 13.5%.

[0161] Example 14

[0162]

[0163]

[0164] Under oil bath conditions at 80°C, 4-methyl-7-aminocoumarin (131 mg, 0.75 mmol), 2-trifluoromethylbenzaldehyde (174 mg, 1.0 mmol), and glacial acetic acid (1 mL) were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol (10 mL) was used as the solvent. The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, and compound 14 was obtained by filtration. The yield of the target product was 57.6%, and the melting range was 277-280°C.

[0165] Example 15

[0166]

[0167] Under oil bath conditions at 80°C, 131 mg (0.75 mmol) of 4-methyl-7-aminocoumarin, 203 mg (1.0 mmol) of 2-fluoro-4-bromobenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol was used as the solvent (10 mL). The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out and was filtered to obtain compound 15. The yield of the target product was 31.1%, and the melting range was 110-111°C.

[0168] Example 16

[0169]

[0170] Under oil bath conditions at 80°C, 131 mg (0.75 mmol) of 4-methyl-7-aminocoumarin, 201 mg (1.0 mmol) of 2-bromo-4-hydroxybenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol was used as the solvent (10 mL). The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, and compound 17 was obtained by filtration. The yield of the target product was 45.7%, and the melting range was 196-201°C.

[0171] Example 17

[0172]

[0173] Under 80°C oil bath conditions, 120 mg (0.75 mmol) of 3-aminocoumarin, 203 mg (1.0 mmol) of 2-fluoro-4-bromobenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. 10 mL of anhydrous methanol was used as the solvent. The reaction was stopped after 8 h. Upon cooling to room temperature, a solid precipitated, which was filtered to obtain compound 17. The yield of the target product was 55.4%, and the melting range was 110-111°C.

[0174] Example 18

[0175]

[0176] Under 80°C oil bath conditions, 120 mg (0.75 mmol) of 3-aminocoumarin, 141 mg (1.0 mmol) of 4-chlorobenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. 10 mL of anhydrous methanol was used as the solvent. The reaction was stopped after 8 h. Upon cooling to room temperature, a solid precipitated, which was filtered to obtain compound 22. The yield of the target product was 45.7%, and the melting range was 196-201°C.

[0177] Example 19

[0178]

[0179] Under 80°C oil bath conditions, 120 mg (0.75 mmol) of 3-aminocoumarin, 185 mg (1.0 mmol) of 4-bromobenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. 10 mL of anhydrous methanol was used as the solvent. The reaction was stopped after 8 h. Upon cooling to room temperature, a solid precipitated, which was filtered to obtain compound 19. The yield of the target product was 45.7%, and the melting range was 196-201°C.

[0180] Example 20

[0181]

[0182] Under oil bath conditions at 80°C, 120 mg (0.75 mmol) of 3-aminocoumarin, 2,6-dibromo-4-pyridinecarboxaldehyde (265 mg, 1.0 mmol) and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol was used as the solvent (10 mL). The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, and compound 20 was obtained by filtration. The yield of the target product was 45.7%, and the melting range was 196-201°C.

[0183] Example 21

[0184]

[0185] Under 80°C oil bath conditions, 120 mg (0.75 mmol) of 3-aminocoumarin, 174 mg (1.0 mmol) of 2-trifluoromethylbenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. 10 mL of anhydrous methanol was used as the solvent. The reaction was stopped after 8 h. Upon cooling to room temperature, a solid precipitated, which was filtered to obtain compound 21. The yield of the target product was 45.7%, and the melting range was 196-201°C.

[0186] Example 22

[0187]

[0188] Under oil bath conditions at 80°C, 120 mg (0.75 mmol) of 3-aminocoumarin, 186 mg (1.0 mmol) of 3-bromo-5-pyridinecarboxaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol was used as the solvent (10 mL). The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, and compound 22 was obtained by filtration. The yield of the target product was 45.7%, and the melting range was 196-201°C.

[0189] Example 23

[0190]

[0191] Under 80°C oil bath conditions, 120 mg (0.75 mmol) of 3-aminocoumarin, 140 mg (1.0 mmol) of 3-chlorobenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. 10 mL of anhydrous methanol was used as the solvent. The reaction was stopped after 8 h. Upon cooling to room temperature, a solid precipitated, which was filtered to obtain compound 23. The yield of the target product was 45.7%, and the melting range was 196-201°C.

[0192] Example 24

[0193]

[0194] Under oil bath conditions at 80°C, 120 mg (0.75 mmol) of 3-aminocoumarin, 173 mg (1.0 mmol) of 2,4-dihydroxy-5-chlorobenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol was used as the solvent (10 mL). The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, and compound 24 was obtained by filtration. The yield of the target product was 45.7%, and the melting range was 196-201°C.

[0195] Example 25

[0196]

[0197] Under oil bath conditions at 80°C, 120 mg (0.75 mmol) of 3-aminocoumarin, 175 mg (1.0 mmol) of 2-trifluoromethyl-3-pyridinecarboxaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol was used as the solvent (10 mL). The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, and compound 25 was obtained by filtration. The yield of the target product was 45.7%, and the melting range was 196-201°C.

[0198] Example 26

[0199]

[0200] Under oil bath conditions at 80°C, 120 mg (0.75 mmol) of 3-aminocoumarin, 176 mg (1.0 mmol) of 2,6-dichloro-3-pyridinecarboxaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol was used as the solvent (10 mL). The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, and compound 26 was obtained by filtration. The yield of the target product was 45.7%, and the melting range was 196-201°C.

[0201] Example 27

[0202]

[0203] Under oil bath conditions at 80°C, 120 mg (0.75 mmol) of 3-aminocoumarin, 209 mg (1.0 mmol) of 2-trifluoromethyl-5-chlorobenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol was used as the solvent (10 mL). The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, and compound 27 was obtained by filtration. The yield of the target product was 60.8%, and the melting range was 120-126°C.

[0204] Example 28

[0205]

[0206] Under 80°C oil bath conditions, 120 mg (0.75 mmol) of 3-aminocoumarin, 242 mg (1.0 mmol) of 3,5-bis(trifluoromethyl)benzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. Anhydrous methanol was used as the solvent (10 mL). The reaction was stopped after 8 h. After cooling to room temperature, a solid precipitated out, and compound 28 was obtained by filtration. The yield of the target product was 66.2%, and the melting range was 125-128°C.

[0207] Example 29

[0208]

[0209] Under 80°C oil bath conditions, 120 mg (0.75 mmol) of 3-aminocoumarin, 136 mg (1.0 mmol) of 2-fluoro-4-bromobenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. 10 mL of anhydrous methanol was used as the solvent. The reaction was stopped after 8 h. Upon cooling to room temperature, a solid precipitated, which was filtered to obtain compound 29. The yield of the target product was 23.1%, and the melting range was 234-239°C.

[0210] Example 30

[0211]

[0212]

[0213] Under 80°C oil bath conditions, 120 mg (0.75 mmol) of 3-aminocoumarin, 125 mg (1.0 mmol) of 2-fluoro-4-bromobenzaldehyde, and 1 mL of glacial acetic acid were sequentially added to a 50 mL round-bottom flask equipped with a stirrer and a condenser. 10 mL of anhydrous methanol was used as the solvent. The reaction was stopped after 8 h. Upon cooling to room temperature, a solid precipitated, which was filtered to obtain compound 30. The yield of the target product was 27.3%, and the melting range was 243-249°C.

[0214] (II) Spectral data of synthesized coumarin compounds

[0215] Using modern spectroscopic techniques, nuclear magnetic resonance hydrogen spectroscopy (NMR) 1 H NMR and carbon nuclear magnetic resonance (NMR) 13 The structure of the synthesized compound was characterized by C NMR.

[0216] Compound 1

[0217] 1 H NMR (400MHz, DMSO-d6) δ10.77(s,1H),10.70(s,1H),7.76(d,J=8.8Hz,1H),6.84(dd,J=8.8,2.4Hz,1H),6.79–6. 68(m,2H),6.03(ddt,J=16.5,10.9,5.7Hz,1H),5.54–5.40(m,1H),5.34(d,J=10.5Hz,1H),4.82(d,J=5.8Hz,2H).

[0218] Compound 2

[0219] 1 1H NMR (600 MHz, Methanol-d4) δ 7.62 (d, J = 8.9 Hz, 1H), 6.63 (dd, J = 8.9, 2.4 Hz, 1H), 6.46 (d, J = 2.3 Hz, 1H), 6.33 (s, 1H), 5.91 (tt, J = 11.5, 5.7 Hz, 1H), 5.30 (dddt, J = 25.1, 11.5, 2.2, 1.0 Hz, 2H), 4.69 (dt, J = 5.8, 0.9 Hz, 2H), 1.29 (s, 3H).

[0220] Compound 3

[0221] 1 1H NMR (600 MHz, Methanol-d4) δ 7.93 (d, J = 8.7 Hz, 1H), 7.11 (d, J = 2.3 Hz, 1H), 7.08 (dd, J = 8.7, 2.3 Hz, 1H), 5.36 (s, 1H), 2.69 (q, J = 6.9 Hz, 1H), 1.88–1.77 (m, 1H), 1.65 (ddd, J = 14.0, 7.6, 6.4 Hz, 1H), 1.29 (d, J = 7.0 Hz, 3H), 1.04 (t, J = 7.5 Hz, 3H).

[0222] 13 13C NMR (151 MHz, MeOD) δ 175.94, 166.40, 158.31, 155.90, 155.33, 124.80, 118.93, 113.48, 111.53, 84.51, 42.27, 27.75, 16.76, 11.84.

[0223] Compound 4

[0224] 1 1H NMR (600 MHz, Methanol-d4) δ 7.93 (dd, J = 13.2, 8.7 Hz, 1H), 7.24 (d, J = 2.3 Hz, 1H), 7.21–7.18 (m, 1H), 3.99 (s, 1H), 2.71 (h, J = 7.0 Hz, 1H), 1.67 (ddd, J = 13.8, 7.5, 6.3 Hz, 1H), 1.30 (dd, J = 7.0, 3.1 Hz, 3H), 1.05 (td, J = 7.5, 1.5 Hz, 3H).

[0225] 13C NMR(151MHz,MeOD)δ175.94,166.42,159.52,158.33,155.93,155.35,150.87 ,124.82,118.94,113.50,111.55,84.51,49.85,42.29,27.76,16.76,11.84.

[0226] Compound 5

[0227] 1 H NMR (600MHz, Methanol-d4) δ7.90(d,J=8.8Hz,1H),7.23(d,J=2.3Hz,1H),7.20(d,J=8.1Hz,2H),4.44(q,J=7.2Hz,2H),2.7 1(h,J=6.9Hz,1H),1.66(ddd,J=13.7,7.5,6.3Hz,1H),1.43(t,J=7.1Hz,3H),1.31(d,J=7.0Hz,4H),1.05(t,J=7.5Hz,3H).

[0228] 13 C NMR(151MHz,MeOD)δ175.80,163.44,160.82,158.05,155.68,155.58,146.97,12 4.67,119.53,113.28,111.87,103.16,64.72,42.28,27.74,16.74,14.21,11.84.

[0229] Compound 6

[0230] 1 H NMR(600MHz, Methanol-d4)δ7.90(d,J=8.8Hz,1H),7.22(d,J=2.3Hz,1H),7.20–7.17(m,1H),3.98(s,2H),1. 83(dp,J=13.4,7.4Hz,1H), 1.66(ddd,J=13.8,7.5,6.3Hz,1H), 1.30(d,J=7.0Hz,3H), 1.04(t,J=7.5Hz,3H).

[0231] 1313C NMR (151 MHz, MeOD) δ 175.58, 174.23, 163.45, 155.58, 155.13, 147.25, 125.53, 124.72, 119.52, 118.95, 113.28, 111.86, 111.55, 103.28, 84.49, 42.28, 27.74, 16.74, 11.84.

[0232] Compound 7

[0233] 1 1H NMR (600 MHz, Methanol-d4) δ 7.93 (d, J = 8.7 Hz, 1H), 7.10 (d, J = 2.2 Hz, 1H), 7.08 (dd, J = 8.7, 2.3 Hz, 1H), 5.36 (s, 1H), 2.76 (h, J = 7.0 Hz, 1H), 1.79 (ddt, J = 13.3, 8.4, 7.4 Hz, 1H), 1.50–1.42 (m, 2H), 1.30 (d, J = 7.0 Hz, 3H), 0.99 (t, J = 7.3 Hz, 3H).

[0234] 13 13C NMR (151 MHz, MeOD) δ 176.09, 166.42, 158.32, 155.89, 155.32, 124.81, 118.92, 113.48, 111.53, 84.48, 40.56, 36.96, 21.44, 17.22, 14.30.

[0235] Compound 8

[0236] 1 1H NMR (600 MHz, Methanol-d4) δ 7.91 (d, J = 8.8 Hz, 1H), 7.22 (d, J = 2.3 Hz, 1H), 7.19 (d, J = 2.5 Hz, 1H), 5.36 (s, 0H), 3.98 (s, 2H), 2.77 (h, J = 6.9 Hz, 1H), 1.86–1.74 (m, 2H), 1.51–1.42 (m, 3H), 1.30 (d, J = 7.0 Hz, 3H), 0.99 (t, J = 7.3 Hz, 4H).

[0237] 13C NMR(151MHz,MeOD)δ176.10,167.54,159.49,158.33,155.93,154.49,150.64,12 4.36,118.91,112.87,111.53,84.52,49.85,40.57,36.97,21.43,17.21,14.28.

[0238] Compound 9

[0239] 1 H NMR(400MHz, Methanol-d4)δ7.90(d,J=8.7Hz,1H),7.09–7.06(m,2H),7.04(d,J=2.3Hz,1H),5.34(s,1H),4.29(q,J=7.1Hz,0 H),2.79–2.68(m,1H),1.83–1.71(m,1H),1.49–1.39(m,3H),1.34–1.27(m,3H),1.27(d,J=2.3Hz,3H),0.97(t,J=7.3Hz,4H). 13 CNMR(101MHz,Methanol-D4)δ176.11,166.44,159.27,158.34,155.91,155.35,148.36,1 24.83,118.94,114.06,111.55,92.76,84.48,40.56,36.98,21.44,17.22,14.30,11.38.

[0240] Compound 10

[0241] 1 H NMR(600MHz,Chloroform-d)δ7.86(d,J=8.2Hz,1H),7.24–7.08(m,2H),5.91(tt,J=11.7,5.8Hz,1H),5.63(s,1H),5.30(dddt,J=25.1,11.5,2.2, 1.0Hz,2H),4.69(dt,J=5.8,1.1Hz,2H),2.55(h,J=7.2Hz,1H),1.71–1.4 8(m,2H),1.43–1.24(m,2H),1.13(d,J=7.3Hz,3H),0.92(t,J=7.3Hz,3H). 13¹³C NMR (151 MHz, Chloroform-d) δ 174.90, 162.37, 161.27, 155.81, 155.31, 154.14, 146.45, 132.65, 128.47, 118.36, 115.26, 112.04, 108.21, 93.21, 66.14, 39.48, 35.52, 20.63, 17.25, 13.92.

[0242] Compound 11

[0243] 1 ¹H NMR (600 MHz, DMSO-d6) δ 10.51 (s, 1H), 7.86 (d, J = 8.9 Hz, 1H), 7.34 (dd, J = 10.7, 2.1 Hz, 1H), 7.28 (dd, J = 8.4, 2.0 Hz, 1H), 7.01 (td, J = 8.6, 1.1 Hz, 1H), 6.77 (dd, J = 8.9, 2.4 Hz, 1H), 6.66 (d, J = 2.4 Hz, 1H). 13 ¹³C NMR (151 MHz, DMSO) δ 163.98, 161.25, 159.78, 153.78, 129.99, 126.96, 126.37, 126.28, 124.63, 118.88, 118.81, 118.26, 118.09, 112.78, 106.38, 101.98.

[0244] Compound 12

[0245] 1 ¹H NMR (600 MHz, Chloroform-d) δ 12.70 (s, 1H), 8.88 (s, 1H), 8.31 (dd, J = 4.4, 1.4 Hz, 1H), 7.68 (d, J = 8.9 Hz, 1H), 7.40 (dd, J = 8.5, 1.4 Hz, 1H), 7.34 (dd, J = 8.5, 4.3 Hz, 1H), 7.29 - 7.28 (m, 1H), 7.27 (d, J = 2.1 Hz, 1H), 6.31 (d, J = 1.4 Hz, 1H), 2.47 (d, J = 1.3 Hz, 3H). 13 ¹³C NMR (151 MHz, Chloroform-d) δ 166.02, 160.69, 158.73, 154.59, 152.00, 150.77, 141.98, 136.97, 127.53, 125.94, 125.37, 119.32, 117.99, 115.10, 109.70, 20.23.

[0246] Compound 13

[0247] 1 H NMR(600MHz,Chloroform-d)δ8.27(d,J=0.8Hz,1H),7.59(d,J=8.4Hz,1H),7.37(d,J=8.5Hz,0H),7.09(d, J=21.7Hz,3H),6.78(s,1H),6.61-6.52(m,1H),6.36(s,1H),6.24(d,J=1.4Hz,1H),2.44(d,J=1.3Hz,3H). 13 C NMR (151MHz, Chloroform-d) δ161.67,154.67,151.88,151.72,149.24,130.90,127.64,118.23,117.84,115.41,112.50,111.76,107.57,19.87.

[0248] Compound 14

[0249] 1 H NMR(600MHz,DMSO-d6)δ8.82(s,1H),7.91(dd,J=6.4,1.5Hz,1H),7.67–7.54(m,3H),7.4 6–7.40(m,2H),7.17(dd,J=8.4,2.2Hz,1H),6.09(q,J=1.4Hz,1H),2.44(d,J=1.3Hz,3H). 13 C10 NMR (151 MHz, Chloroform-d) δ 162.71, 161.67, 154.67, 151.72, 150.75, 132.88, 132.13, 131.24, 129.99, 129.95, 127.64, 126.89, 125.14, 118.24, 117.75, 111.76, 107.44, 19.87. Compound 15

[0250] 1 H NMR (600MHz, DMSO-d6) δ7.84–7.80(m,1H),7.78(t,J=8.0Hz,1H),7.63(dd,J=8.3,1.8Hz,1H),7.40(d,J= 8.6Hz,1H),6.56(dd,J=8.6,2.2Hz,1H),6.43–6.36(m,1H),5.90(q,J=1.2Hz,1H),2.30(d,J=1.2Hz,3H). 13C NMR(151MHz,DMSO)δ163.74,162.01,160.71,155.44,153.72,153.05,130.84,1 28.55,126.28,126.17,120.31,119.72,118.06,117.87,113.62,111.15,17.97.

[0251] Compound 16

[0252] 1 H NMR (600MHz, MeOD) δ8.91(s,1H),8.71(s,1H),7.59(dd,J=17.9,8.5Hz,2H),7.40(d,J=2.3Hz,1H),7.16(d,J=1. 9Hz, 1H), 7.10 (dd, J = 8.3, 2.3Hz, 1H), 6.81 (dd, J = 8.7, 1.9Hz, 1H), 6.09 (q, J = 1.4Hz, 1H), 2.44 (d, J = 1.3Hz, 3H). 13 C NMR(151MHz,MeOD)δ162.48,160.70,159.44,153.72,153.05,150.83,137.38,1 26.18,124.68,123.86,117.40,116.59,114.02,111.15,108.83,107.45,17.98.

[0253] Compound 17

[0254] 1 H NMR(600MHz,DMSO-d6)δ7.82–7.78(m,1H),7.78–7.74(m,1H),7.63(dt,J=8.3,1.2Hz,1H),7.61–7 .58(m,1H),7.42–7.39(m,1H),7.28–7.25(m,1H),7.23(dd,J=7.2,1.8Hz,1H),7.21–7.18(m,1H). 13 C NMR(151MHz,DMSO)δ163.74,162.01,158.60,157.46,147.88,133.27,130.83,13 0.81,128.52,125.31,124.78,124.46,121.77,120.30,120.14,115.39,107.65.

[0255] Compound 18

[0256] 1 H NMR (600MHz, DMSO-d6) δ9.00(s,1H),7.81(s,1H),7.79–7.72(m,3H),7.66–7.61(m,2H),7.54–7.47(m,1H),7.35–7.28(m,2H). 13 C NMR (151MHz, DMSO) δ163.44,159.22,151.75,136.89,135.42,132.92,132.73,129.82,129.41,129.28,128.38,125.33,121.92,118.21.

[0257] Compound 19

[0258] 1 H NMR(600MHz,DMSO-d6)δ9.00(s,1H),7.81(s,1H),7.76(dd,J=7.6,1.6Hz,1H) ,7.74–7.69(m,2H),7.61–7.55(m,2H),7.54–7.47(m,1H),7.35–7.28(m,2H). 13 C NMR (151MHz, DMSO) δ163.44,159.32,151.75,135.84,132.92,132.73,131.82,129.50,129.28,128.38,125.33,124.42,121.92,118.21.

[0259] Compound 20

[0260] 1 H NMR(600MHz,DMSO-d6)δ8.80(s,2H),7.83(dd,J=7.8,1.6Hz,1H),7.66–7.62(m, 1H),7.47(d,J=8.3Hz,1H),7.37–7.35(m,2H),7.31(ddd,J=7.6,5.7,2.8Hz,1H). 13 CNMR(151MHz,DMSO)δ191.20,160.82,158.45,157.90,151.17,133.13,130.25,128.82,125.99,124.69,124.31,121.06,119.28,107.51.

[0261] Compound 21

[0262] 11H NMR (600 MHz, DMSO-d6) δ 10.28 (s, 1H), 8.12–8.10 (m, 1H), 7.96–7.94 (m, 1H), 7.94–7.91 (m, 1H), 7.91 (d, J = 3.0 Hz, 1H), 7.47–7.41 (m, 1H), 7.41–7.37 (m, 1H), 7.27 (dd, J = 7.9, 1.6 Hz, 1H), 7.23 (dd, J = 7.2, 1.8 Hz, 1H), 7.20 (dd, J = 7.3, 5.7 Hz, 1H). 13 13C NMR (151 MHz, DMSO) δ 189.50, 158.45, 147.73, 134.16, 133.13, 130.52, 126.42, 126.38, 126.34, 125.16, 124.64, 124.63, 124.30, 121.62, 115.74, 115.24, 107.49.

[0263] Compound 22

[0264] 1 1H NMR (600 MHz, DMSO-d6) δ 10.07 (s, 1H), 9.05 (d, J = 1.7 Hz, 1H), 9.00 (d, J = 2.3 Hz, 1H), 8.47 (t, J = 2.1 Hz, 1H), 7.41 (dd, J = 7.2, 2.0 Hz, 1H), 7.27 (dd, J = 7.8, 1.7 Hz, 1H), 7.23 (dd, J = 7.2, 1.8 Hz, 1H), 7.21–7.19 (m, 1H), 6.71 (s, 1H). 13 13C NMR (151 MHz, DMSO) δ 191.46, 158.74, 155.16, 149.46, 148.02, 138.64, 133.41, 132.75, 125.46, 124.92, 124.60, 121.90, 121.02, 115.53, 107.80.

[0265] Compound 23

[0266] 1 1H NMR (600 MHz, DMSO-d6) δ 9.03 (s, 1H), 7.83–7.75 (m, 2H), 7.75–7.69 (m, 2H), 7.54–7.48 (m, 1H), 7.48–7.42 (m, 2H), 7.35–7.28 (m, 2H). 13C NMR (151MHz, DMSO) δ163.44,158.84,151.75,139.48,134.87,132.92,132.7 3,130.97,129.28,129.23,128.38,128.16,126.62,125.33,121.92,118.21.

[0267] Compound 24

[0268] 1 H NMR(600MHz,DMSO-d6)δ11.37(s,1H),10.86(s,1H),9.97(s,1H),7.60(s,1H),7.44–7.41(m,1H),7.39( dd,J=16.8,1.6Hz,1H),7.28–7.25(m,1H),7.24–7.21(m,1H),7.19(dd,J=7.3,5.7Hz,1H),6.71(s,1H). 13 C NMR(151MHz,DMSO)δ188.74,158.73,148.02,138.42,135.39,133.95,133.4 1,130.76,128.94,125.45,124.92,124.60,121.90,116.08,115.53,107.78.

[0269] Compound 25

[0270] 1 H NMR (600MHz, DMSO-d6) δ8.58(ddd,J=9.6,7.5,2.1Hz,1H),8.45(dd,J=4.8,2.0Hz,1H),8.00(s,1H),7.77(dd,J=7.7,1.6Hz,1H) ,7.61(ddd,J=8.7,7.3,1.6Hz,1H),7.56(ddd,J=6.9,4.9,1.4Hz,1H),7.41(dd,J=7.3,1.9Hz,1H),7.38(dd,J=7.5,1.1Hz,1H). 13 C NMR(151MHz,DMSO)δ163.39,157.53,151.75,150.64,146.02,134.32,133.00,132.92,13 2.81,129.28,128.30,125.69,125.33,124.75,123.55,121.92,121.40,119.26,118.21.

[0271] Compound 26

[0272] 1 H NMR(600MHz,DMSO-d6)δ8.56(d,J=5.4Hz,1H),7.80–7.75(m,1H),7.43–7.42(m,1H),7. 41(d,J=2.0Hz,1H),7.29–7.25(m,1H),7.24(dd,J=7.2,1.9Hz,1H),7.22–7.18(m,1H). 13 C NMR (151MHz, DMSO) δ158.58,152.81,152.04,151.34,150.88,147.87,145.48,133.26,126.86,125.98,125.30,124.77,124.44,121.75,115.38.

[0273] Compound 27

[0274] 1 H NMR(600MHz,DMSO-d6)δ9.50(q,J=2.1Hz,1H),8.15–8.08(m,1H),7.97(t,J=1.3Hz,1H),7.86(dd,J=8.6,2.3Hz,1H),7.8 1(dd,J=7.8,1.6Hz,1H),7.64(ddd,J=8.8,7.3,1.6Hz,1H),7.47(d,J=8.3Hz,1H),7.42–7.40(m,1H),7.29–7.24(m,1H). 13 C NMR (151MHz, DMSO) δ188.94,161.35,159.96,158.61,147.89,133.28,130.35,12 8.29,125.32,124.78,124.46,121.77,116.02,115.39,112.25,107.67,103.55.

[0275] Compound 28

[0276] 1 H NMR(600MHz,DMSO-d6)δ10.16(s,1H),9.26(s,1H),8.57(d,J=32.0Hz,1H),8.04(s,1H) ,7.40(dd,J=7.5,1.7Hz,1H),7.26(dd,J=7.8,1.7Hz,1H),7.21(dd,J=8.2,1.9Hz,1H). 13C NMR (151MHz, DMSO) δ191.29,161.25,157.60,152.23,148.16,138.31,135.10,133.5 5,132.61,128.90,125.59,125.18,125.05,124.73,119.77,116.25,115.66,107.92.

[0277] Compound 29

[0278] 1 H NMR(600MHz,DMSO-d6)δ7.95(dd,J=8.6,0.6Hz,1H),7.91(s,1H),7.74(dd,J=7.8,1.6Hz,1H),7.61 –7.59(m,1H),7.58(ddd,J=6.2,4.8,2.5Hz,2H),7.42–7.40(m,2H),7.28–7.25(m,3H),3.95(s,3H). 13 C NMR (151MHz, DMSO) δ163.03,158.86,158.60,147.88,145.71,138.53,133.27,125.32,124.78,124.45,123.53,121.76,120.55,115.39,56.15.

[0279] Compound 30

[0280] 1 H NMR(600MHz,DMSO-d6)δ9.25(s,1H),8.47(dd,J=4.8,2.2Hz,1H),8.11–8.04(m,1H),7.81(s,1H) ,7.76(dd,J=7.6,1.6Hz,1H),7.54–7.47(m,1H),7.43(dd,J=8.2,4.9Hz,1H),7.35–7.28(m,2H). 13 C NMR(151MHz,DMSO)δ158.59,157.42,155.97,151.89,147.88,138.82,138.8 0,135.30,133.27,128.61,125.31,124.77,124.45,121.76,115.91,115.38.

[0281] (III) Antifungal activity of coumarin compounds

[0282] 1. Test bacteria used in the experiment

[0283]

[0284] The above four standard bacterial strains were used in this part of the experiments. All strains were provided by the Chemical Ecology Laboratory of the Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, and can also be purchased directly from Gary Chemical Network, Shanghai Boco Biotechnology Co., Ltd., etc.

[0285] 2. Preparation of PDA culture medium

[0286] (1) Weighing and cooking: Calculate the total amount of culture medium required based on the requirement of 10 mL of PDA medium per petri dish. Weigh a certain amount of peeled potatoes according to the calculation of adding 200 g of potatoes to every 1000 mL of distilled water. Cut the potatoes into small pieces and put them in a pot. Add 1000 mL of water and heat to a boil in the pot. Cook the potatoes until soft but not mushy. Filter while hot through 6-8 layers of gauze and discard the residue. Add water to the filtrate to 1000 mL.

[0287] (2) Heating to dissolve: Put the filtrate into a pot, add glucose (20g / 1000mL distilled water) and agar powder (18g / 1000mL distilled water), heat over low heat and stir constantly with a glass rod to prevent the agar powder from sticking to the bottom or overflowing. After the agar is completely dissolved, add water to the required amount.

[0288] (3) Dispensing and autoclaving: Dispense the prepared culture medium into 500ml Erlenmeyer flasks. It is advisable to dispense into the Erlenmeyer flasks no more than half full of their volume, seal them with sealing film, and autoclave them for later use.

[0289] 3. Preparation of compound solutions for synthesized coumarin compounds

[0290] The compound solution was prepared to a concentration of 100 μg / mL by accurately weighing 12.5 mg of the compound into a centrifuge tube, adding 1250 μL of dimethyl sulfoxide (DMSO) using a pipette, and sonicating until completely dissolved. The compound solution was then sterilized under a UV lamp for 30 min.

[0291] 4. Experimental Procedure

[0292] Place 10 mL of PDA medium in a petri dish, add 100 μL of the compound solution, gently shake to mix, label, and cool horizontally. Use 100 μL of dimethyl sulfoxide (DMSO) as a blank control. Perform three parallel experiments. Use commercially available fungicides such as carbendazim and chlorothalonil as positive controls. Use a 0.7 cm diameter punch to collect vigorous mycelial cakes and punch concentric circles. Inoculate the center of fresh medium and invert the dish in a 25°C incubator. Measure the colony diameter after 96 hours using the cross-hatching method, calculate the average diameter, and determine the inhibitory rate of the compound.

[0293] Formula for calculating antibacterial rate: I = [(D0 - D] t [(D0-0.7)]×100%

[0294] (I: Mycelial growth inhibition rate, D0: Diameter of blank colony, D) t (Diameter of colonies after chemical treatment)

[0295] 5. Measurement Results

[0296] The inhibitory activity of the synthetic coumarin compounds against four common plant pathogens—Botrytis cinerea, Alternaria solanacea, Fusarium oxysporum, and Alternaria alternata—was determined using the mycelial growth rate method described above. The experimental results are shown in Table 1.

[0297] Table 1

[0298]

[0299]

[0300] Note: Higher values ​​in Table 1 indicate higher antibacterial activity, while negative values ​​indicate that the compound promotes fungal growth.

[0301] In this invention, the antifungal activity of coumarin compounds was studied using four common plant pathogenic fungi: Botrytis cinerea, Alternaria solani, Fusarium oxysporum, and Alternaria alternata. The mycelial growth rate method was used to screen for pathogenic fungi, which can lay the foundation for the development of novel plant-derived pesticides.

[0302] As shown in Table 1, at a mass concentration of 100 μg / mL, compounds 1-30 synthesized in this invention exhibit varying degrees of antifungal activity against the four tested plant pathogenic fungi, demonstrating their potential as novel plant-derived pesticide antifungal agents. It should be noted that carbendazim or chlorothalonil have significant structural differences from the coumarin compounds of this invention and are not used for comparison of their antifungal rates with the compounds synthesized in this patent. Using carbendazim and chlorothalonil as positive controls sufficiently demonstrates the reliability of this experiment.

[0303] The present invention has been described in detail above with reference to specific embodiments and exemplary examples; however, these descriptions should not be construed as limiting the present invention. Those skilled in the art will understand that various equivalent substitutions, modifications, or improvements can be made to the technical solutions and embodiments of the present invention without departing from the spirit and scope of the invention, and all such modifications and improvements fall within the scope of the present invention. The scope of protection of the present invention is defined by the appended claims.

[0304] All publications, patent applications, patents, and other references mentioned in this specification are incorporated herein by reference. Unless otherwise defined, all technical and scientific terms used in this specification have the meanings commonly understood by those skilled in the art. In case of conflict, the definitions in this specification shall prevail.

[0305] When this specification uses the prefixes “known to those skilled in the art,” “prior art,” or similar terms to derive materials, substances, methods, steps, apparatus, or components, the objects derived from such prefixes cover those commonly used in the art at the time of this application’s filing, but also include those that are not currently commonly used but will become generally recognized in the art as suitable for similar purposes.

[0306] In the context of this specification, except where expressly stated otherwise, any matters or issues not mentioned shall apply directly to those known in the art without any modification.

Claims

1. A coumarin compound, characterized in that, The coumarin compounds are selected from the following compounds: .

2. A coumarin compound, characterized in that, The coumarin compounds are , .

3. A coumarin compound, characterized in that, The coumarin compounds are .

4. A coumarin compound, characterized in that, The coumarin compounds are selected from the following compounds: 、 。 5. The use of a coumarin compound according to any one of claims 1-4 in the preparation of a pesticide antibacterial agent against at least one of the pathogens Alternaria solani and Alternaria alternata.

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