Multi-heterocycle spliced vanillin compound as well as preparation method and application thereof

Abstract

The invention provides a vanillin compound spliced by polyheterocycles. The polyheterocycles are one of a benzimidazole ring, a thiadiazole ring, an oxadiazole ring and a thiazole ring; the vanillin compound spliced by polyheterocycles is one of vanillin derivatives containing a benzimidazole ring, a thiadiazole ring, an oxadiazole ring and a thiazole ring, and the chemical general formula is shown in the specification. The polyheterocycle spliced vanillin compound is prepared through a condensation reaction and a condensation-cyclization reaction, and the compound has good antibacterial activity on valsa mali, red kidney bean root rot, fusarium graminearum, millet pyricularia grisea, botrytis cinerea, alternaria solani and watermelon fusarium oxysporum. The compound can be used for preparing a novel efficient antibacterial agent and is applied to prevention and treatment of the pathogenic bacteria.

Description

Technical Field

[0001] This invention belongs to the field of vanillin compounds, specifically relating to vanillin compounds with multiple heterocyclic structures, their preparation methods, and applications. Background Technology

[0002] Vanillin is a compound mainly derived from the orchid plant Vanilla sylvestris (Vanilla vulgaris). Vanilla planifolia Natural flavoring substances isolated from [various sources] have attracted much attention due to their various biological functions, including antibacterial, anti-inflammatory, antioxidant, and anticancer properties. Among them, the literature [Jing Y, Chen YZ, Wu YX, et al. Inhibitory effects and mechanisms of vanillin on gray mold and black rot of cherry tomatoes[J]. PesticideBiochemistry and Physiology,2021, 175: 104859.] and [Zhang Xuemin. Study on the control mechanism of vanillin against apple rot[D]. Liaoning University,2021.] reported the antibacterial activity of vanillin against various fungi, including those causing gray mold, black rot, and apple rot in cherry and tomato. The literature [Yan Xuan. The role of vanillin in a neuroinflammation-mediated Parkinson's disease model[D]. Changchun: Jilin University,2017.] reported that vanillin has good anti-inflammatory activity. The literature [Liu Junhai, Li Zhizhou, Wang Junhong. Study on vanillin extracted from vanilla pods by microwave and its antioxidant activity [J]. Food Science and Technology, 2016, (8): 210-216] and [Yang Qingming, Ding Lan, Yang Hong, et al. Study on antioxidant activity of edible flavoring vanillin [J]. Food Research and Development, 2007, 28(1): 85-88.] reported that vanillin has good antioxidant activity. The literature [Lirdprapamongkol K, Sakurai H, Kawasaki N, et al. Vanillin suppresses invitro invasion and invivo 57 metastasis of mouse breast cancer cells [J]. European Journal of Pharmaccutical Sciences, 2005, 25(1): 57-65.] reported that vanillin has a significant inhibitory effect on mouse 4T1 cancer cells.

[0003] Through the previous literature research, it is found that vanillin has less application in agriculture, and only a few reports of vanillin agricultural antibacterial are reported. However, the biological activity is relatively mild, the antibacterial effect is poor, and it cannot be applied to actual production, and some derivatives with greatly improved antibacterial activity of vanillin need to be developed. SUMMARY

[0004] The purpose of the present application is to solve the problem of weak antibacterial activity of vanillin in the prior art, which cannot be applied to actual production.

[0005] To solve the above technical problems, the present application provides a multi-heterocyclic spliced vanillin compound and its preparation method and application.

[0006] The technical solution adopted by the present application is as follows: The multi-heterocyclic spliced vanillin compound is one of vanillin derivatives containing benzimidazole ring, thiadiazole ring, oxadiazole ring and thiazole ring, and the chemical general formula of the multi-heterocyclic spliced vanillin compound is as shown in the following formula: In the chemical general formula: In the formula, compounds (1)-(7) are benzimidazole vanillin derivatives, (8)-(14) are thiadiazole vanillin derivatives, (15)-(24) are oxadiazole vanillin derivatives, and (25)-(33) are thiazole vanillin derivatives.

[0007] Preferably, the thiadiazole ring is 1,3,4-thiadiazole ring, and the oxadiazole ring is 1,3,4-oxadiazole ring.

[0008] The preparation method of the multi-heterocyclic spliced vanillin compound comprises the following steps: condensation reaction and condensation-cyclization reaction.

[0009] Further, the condensation reaction comprises A1 or A2, and the condensation-cyclization reaction comprises one of B1, B2, B3 and B4.

[0010] A1: Take 0.8 mmol of 2-(chloromethyl)-5-phenyl-1,3,4-oxadiazole and 1.0 mmol of vanillin are dissolved in 2 mL of acetone, 1.2 mmol of potassium carbonate and 0.1 mmol of potassium iodide are added, the reaction is stirred at 55 ℃, the reaction progress is tracked, after the reaction is completed, the filtrate is concentrated under reduced pressure, and preparative thin layer chromatography on silica gel is used for separation to obtain compound 3-methoxy-4-[ (5-phenyl-1,3,4-oxadiazol-2-yl)methoxy]benzaldehyde.

[0011] A2: Take 1.4 mmol of vanillin, 1.1 mmol of compound 2-(chloromethyl)-1H-benzimidazole, 1.7 mmol of potassium carbonate and 0.1 mmol of potassium iodide are dissolved in 2 mL of acetone; the reaction is stirred at 55 ℃, the reaction progress is tracked, after the reaction is completed, the filtrate is concentrated under reduced pressure, and preparative thin layer chromatography on silica gel is used for separation to obtain compound 2-[ (3-methoxy-4-formylphenoxy)methyl]-1H-benzimidazole.

[0012] B1: Take 0.4 mmol of 3-methoxy-4-[ (5-phenyl-1,3,4-oxadiazol-2-yl)methoxy]benzaldehyde, 0.4 mmol of substituted o-phenylenediamine and 0.4 mmol of sodium metabisulfite are dissolved in 2 mL of dimethyl sulfoxide, the reaction is stirred at 140 ℃, the reaction progress is tracked, after the reaction is completed, the reaction solution is cooled to room temperature, 10 mL of water is added to quench the reaction, 40 mL of ethyl acetate is used for extraction for 5 times, the organic phase is combined, dried with anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and preparative thin layer chromatography on silica gel is used for separation.

[0013] B2: Take 0.4 mmol of substituted aromatic hydrazine and 0.4 mmol of 3-methoxy-4-[ (5-phenyl-1,3,4-oxadiazol-2-yl)methoxy]benzaldehyde are dissolved in 2 mL of methanol, the reaction is stirred at 65 ℃, the reaction progress is tracked, after the reaction is completed, it is concentrated under reduced pressure, 2 mL of toluene is added at room temperature to dissolve the above residue, then 0.5 mmol of Lawesson's reagent and 0.5 mmol of 4-(dimethylamino)pyridine are added in sequence to the above mixture, the reaction is stirred at 110 ℃, the reaction progress is tracked, after the reaction is completed, it is concentrated under reduced pressure, and preparative thin layer chromatography on silica gel is used for separation.

[0014] B3: Take 0.6 mmol of substituted aromatic hydrazine and 0.4 mmol of compound 2-[(3-methoxy-4-formylphenoxy)methyl]-1H-benzimidazole dissolved in 2 mL of methanol, stirred at 65 °C, track the reaction process, after the reaction is completed, concentrated under reduced pressure, take 2 mL of dichloromethane-methanol to dissolve the above residue, then add 0.5 mmol of diethyl iodo benzene to the above mixture, stirred at room temperature, track the reaction process, after the reaction is completed, concentrated under reduced pressure, prepare silica gel thin layer chromatography, separation.

[0015] B4: Take 0.5 mmol of compound 2-[(3-methoxy-4-formylphenoxy)methyl]-1H-benzimidazole and 0.8 mmol of substituted 4-phenyl-thiazol-2-amine dissolved in 2 mL of toluene, stirred at 110 °C, track the reaction process, after the reaction is completed, concentrated under reduced pressure; take 2 mL of anhydrous ethanol to dissolve the above residue, then take 3.2 mmol of sodium borohydride in batches and dissolve in the above mixture, stirred at room temperature, track the reaction process, after the reaction is completed, concentrated under reduced pressure, adjust the pH value to 7 with saturated sodium bicarbonate solution, extract 3 times with 40 mL of ethyl acetate, combine the organic phase, dry the organic phase with anhydrous sodium sulfate; filter, concentrate under reduced pressure, prepare silica gel thin layer chromatography for separation.

[0016] Further, compounds (1)-(7) are obtained by condensation reaction Al and condensation-cyclization reaction Bl; compounds (8)-(14) are obtained by condensation reaction Al and condensation-cyclization reaction B2; compounds (15)-(24) are obtained by condensation reaction A2 and condensation-cyclization reaction B3; compounds (25)-(33) are obtained by condensation reaction A2 and condensation-cyclization reaction B4.

[0017] Preferably, the substituted o-phenylenediamine is one of 4-methyl-o-phenylenediamine, 3,4-dimethyl-o-phenylenediamine, 4-chloro-o-phenylenediamine, 4-bromo-o-phenylenediamine, 4-nitro-o-phenylenediamine or 4-trifluoromethyl-o-phenylenediamine.

[0018] Preferably, the substituted aromatic hydrazide is one of 3-methyl aromatic hydrazide, 4-methyl aromatic hydrazide, 3-methoxy aromatic hydrazide, 4-tert-butyl aromatic hydrazide, 2-furan aromatic hydrazide, 2-thiophene aromatic hydrazide, 4-fluoro aromatic hydrazide, 2-chloro aromatic hydrazide or 4-bromo aromatic hydrazide.

[0019] Preferably, the substituted benzhydrazide is one of 2-methoxy benzhydrazide, 4-methyl benzhydrazide, 4-methoxy benzhydrazide, 4-fluoro benzhydrazide, 4-chloro benzhydrazide, 3-bromo benzhydrazide, 4-bromo benzhydrazide, 3,4,-dichloro benzhydrazide or 2-nitro benzhydrazide.

[0020] The vanillin compound with multi-heterocycle splicing can be applied in the preparation of agricultural antibacterial agents, and is applied in the prevention and treatment of Valsa mali, Rhizoctonia bataticola, Gibberella zeae, Botrytis cinerea, Alternaria solani and Fusarium oxysporum.

[0021] The vanillin compound with multi-heterocycle splicing has good antibacterial activity on Valsa mali, Valsa ceratosperma , Rhizoctonia bataticola, Fusarium solani , Gibberella zeae, Fusarium graminearum , Botrytis cinerea, Pyricularia grisea , Alternaria solani, Botrytis cinerea , and Fusarium oxysporum, Alternaria , which indicates that introducing benzimidazole, thiazole, thiazole or oxadiazole heterocycle structure into the vanillin molecular skeleton helps to improve the antibacterial activity of vanillin. solani The antibacterial activity of the compound (10) on Valsa mali is as high as 100%, which is much higher than that of the raw material vanillin, and shows strong antibacterial effect. Fusarium oxysporum Therefore, the vanillin compound with multi-heterocycle splicing can be applied to prepare a new type of high-efficiency antibacterial agent. DETAILED DESCRIPTION

[0022] The application will be further described below in combination with specific examples.

[0023] Vanillin compound with multi-heterocycle splicing The vanillin compound with multi-heterocycle splicing is one of vanillin derivatives containing benzimidazole ring, thiazole ring, oxadiazole ring and thiazole ring, and the chemical general formula of the vanillin compound with multi-heterocycle splicing is shown in the following formula: In the chemical general formula, Compounds (1)-(7) are benzimidazole vanillin derivatives, (8)-(14) are thiazole vanillin derivatives, (15)-(24) are oxadiazole vanillin derivatives, and (25)-(33) are thiazole vanillin derivatives.

[0024] Preferably, the thiazole ring is a 1,3,4-thiazole ring, and the oxadiazole ring is a 1,3,4-oxadiazole ring.

[0025] Method for preparing polyheterocycle-spliced vanillin compounds The method for preparing polyheterocycle-spliced vanillin compounds comprises the following steps: a condensation reaction and a condensation-cyclization reaction.

[0026] Further, the condensation reaction comprises A1 or A2, and the condensation-cyclization reaction comprises one of B1, B2, B3, and B4.

[0027] A1: 0.8 mmol of 2-(chloromethyl)-5-phenyl-1,3,4-oxadiazole and 1.0 mmol of vanillin are dissolved in 2 mL of acetone, 1.2 mmol of potassium carbonate and 0.1 mmol of potassium iodide are added, and the reaction is stirred at 55°C. The reaction progress is tracked, and after the reaction is completed, the filtrate is concentrated under reduced pressure. Compound 3-methoxy-4-[ (5-phenyl-1,3,4-oxadiazol-2-yl) methoxy] benzaldehyde is obtained by silica gel thin layer chromatography separation.

[0028] A2: 1.4 mmol of vanillin, 1.1 mmol of compound 2-(chloromethyl)-1H-benzimidazole, 1.7 mmol of potassium carbonate, and 0.1 mmol of potassium iodide are dissolved in 2 mL of acetone; the reaction is stirred at 55°C. The reaction progress is tracked, and after the reaction is completed, the filtrate is concentrated under reduced pressure. Compound 2-[ (3-methoxy-4-formylphenoxy) methyl]-1H-benzimidazole is obtained by silica gel thin layer chromatography separation.

[0029] B1: 0.4 mmol of 3-methoxy-4-[ (5-phenyl-1,3,4-oxadiazol-2-yl) methoxy] benzaldehyde, 0.4 mmol of substituted o-phenylenediamine, and 0.4 mmol of sodium metabisulfite are dissolved in 2 mL of dimethyl sulfoxide. The reaction is stirred at 140°C. The reaction progress is tracked, and after the reaction is completed, the reaction solution is cooled to room temperature. 10 mL of water is added to quench the reaction, and the organic phase is extracted with 40 mL of ethyl acetate 5 times. The organic phase is dried with anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and separated by silica gel thin layer chromatography.

[0030] B2: Take 0.4 mmol of substituted aromatic hydrazine and 0.4 mmol of 3-methoxy-4-[(5-phenyl-1,3,4-oxadiazol-2-yl)methoxy]benzaldehyde dissolved in 2 mL of methanol, stirred at 65 ℃, track the reaction process, after the reaction is completed, concentrated under reduced pressure, room temperature, add 2 mL of toluene to dissolve the above residue, then take 0.5 mmol of Lawesson's reagent and 0.5 mmol of 4-(dimethylamino)pyridine to the above mixture, stirred at 110 ℃, track the reaction process, after the reaction is completed, concentrated under reduced pressure, prepare silica gel thin layer chromatography, separation.

[0031] B3: Take 0.6 mmol of substituted aromatic hydrazine and 0.4 mmol of 2-[(3-methoxy-4-formylphenoxy)methyl]-1H-benzimidazole dissolved in 2 mL of methanol, stirred at 65 ℃, track the reaction process, after the reaction is completed, concentrated under reduced pressure, take 2 mL of dichloromethane-methanol to dissolve the above residue, then add 0.5 mmol of iodonium benzyl diethyl to the above mixture, stirred at room temperature, track the reaction process, after the reaction is completed, concentrated under reduced pressure, prepare silica gel thin layer chromatography, separation.

[0032] B4: Take 0.5 mmol of 2-[(3-methoxy-4-formylphenoxy)methyl]-1H-benzimidazole and 0.8 mmol of substituted 4-phenyl-thiazol-2-amine dissolved in 2 mL of toluene, stirred at 110 ℃, track the reaction process, after the reaction is completed, concentrated under reduced pressure; take 2 mL of anhydrous ethanol to dissolve the above residue, then take 3.2 mmol of sodium borohydride in batches to the above mixture, stirred at room temperature, track the reaction process, after the reaction is completed, concentrated under reduced pressure, adjust the pH value to 7 with saturated sodium bicarbonate solution, extract 3 times with 40 mL of ethyl acetate, combine the organic phase, dry the organic phase with anhydrous sodium sulfate; filter, concentrate under reduced pressure, prepare silica gel thin layer chromatography for separation.

[0033] Compounds (1)-(7) are obtained by condensation reaction A1 and condensation-cyclization reaction B1; compounds (8)-(14) are obtained by condensation reaction A1 and condensation-cyclization reaction B2; compounds (15)-(24) are obtained by condensation reaction A2 and condensation-cyclization reaction B3; compounds (25)-(33) are obtained by condensation reaction A2 and condensation-cyclization reaction B4.

[0034] Preferably, the substituted o-phenylenediamine is one of 4-methyl-o-phenylenediamine, 3,4-dimethyl-o-phenylenediamine, 4-chloro-o-phenylenediamine, 4-bromo-o-phenylenediamine, 4-nitro-o-phenylenediamine or 4-trifluoromethyl-o-phenylenediamine.

[0035] Preferably, the substituted aromatic hydrazide is one of 3-methyl aromatic hydrazide, 4-methyl aromatic hydrazide, 3-methoxy aromatic hydrazide, 4-tert-butyl aromatic hydrazide, 2-furan aromatic hydrazide, 2-thiophene aromatic hydrazide, 4-fluoro aromatic hydrazide, 2-chloro aromatic hydrazide or 4-bromo aromatic hydrazide.

[0036] Preferably, the substituted benzhydrazide is one of 2-methoxy benzhydrazide, 4-methyl benzhydrazide, 4-methoxy benzhydrazide, 4-fluoro benzhydrazide, 4-chloro benzhydrazide, 3-bromo benzhydrazide, 4-bromo benzhydrazide, 3,4,-dichloro benzhydrazide or 2-nitro benzhydrazide.

[0037] Physicochemical properties of polyheterocyclic spliced vanillin compounds The physicochemical properties of compound (1) are as follows: 1) Brown-yellow solid, melting point 151.1-151.8℃; 2) The nuclear magnetic resonance spectrum characteristics of the compound are as follows: In deuterated DMSO as solvent, TMS as internal standard, wherein the peaks are as follows: 1 H NMR (400 MHz, DMSO- d 6): δ =8.03–8.05 (m, 2H), 7.85 (d, J = 2.0 Hz, 1H), 7.78 (dd, J = 2.0, 8.4 Hz, 1H),7.58–7.67 (m, 6H),7.37 (d, J = 8.4 Hz, 1H), 7.19–7.21 (m, 2H), 5.55 (s, 2H),3.91 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ = 165.8, 163.5, 152.0, 150.1, 148.9,133.3, 130.5, 127.6, 125.5, 124.0, 122.9, 120.0, 115.8, 111.3, 61.5, 56.7. 3) The high-resolution mass spectrum characteristics of the compound are as follows: Using electrospray ionization: calcd for C 23 H 19 N4O3 + , ([M+H] + ) 399.1457, found399.1456.。

[0038] The physicochemical properties of compound (2) are as follows: 1) Brown solid, melting point 116.3-116.8℃; 2) The nuclear magnetic resonance spectrum characteristics of the compound are as follows: In deuterated DMSO as solvent, TMS as internal standard, wherein the peaks are as follows: 1 H NMR (400 MHz, DMSO- d 6): δ =12.64 (s, 1H), 8.04 (d, J = 7.2 Hz, 2H), 7.81 (s, 1H), 7.73 (d, J = 8.4 Hz, 1H),7.33–7.66 (m, 6H), 7.01 (d, J = 8.0 Hz, 1H), 5.53 (s, 2H), 3.90 (s, 3H), 2.42(s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ = 165.8, 163.5, 150.4, 148.8, 132.3,130.5, 127.6, 125.8, 124.0, 119.8, 115.8, 111.1, 61.5,56.7, 22.3. 3) The high resolution mass spectrum characteristics of the compound are as follows: Using electrospray ionization: calcd for C 24 H 21 N4O3 + , ([M+H] + ) 413.1614, found413.1618.。

[0039] The physicochemical properties of compound (3) are as follows: 1) White solid, melting point 123.8-124.5℃; 2) The nuclear magnetic resonance spectrum characteristics of the compound are as follows: In deuterated DMSO as solvent, TMS as internal standard, wherein the peaks are as follows: 1 H NMR (400 MHz, DMSO- d 6): δ =12.53 (s, 1H), 8.02–8.05 (m, 2H), 7.80 (d, J= 2.0 Hz, 1H), 7.61–7.72 (m, 4H),7.27–7.40 (m, 3H), 5.53 (s, 2H), 3.90 (s, 3H), 2.32 (s, 6H). 13 C NMR (100 MHz, DMSO- d 6): δ = 165.8, 163.6, 151.1, 150.4, 148.6, 143.4, 134.4, 133.3, 131.8,130.7, 130.5,127.6, 126.0, 123.4, 119.7, 115.9, 112.1, 111.1, 61.5, 56.7,21.0. 3)The high resolution mass spectrum of the compound is characterized as follows: The electrospray ionization is used: calcd for C 25 H 22 N4O3 + , ([M+H] + ) 427.1770, found427.1777.。

[0040] The physicochemical properties of compound (4) are as follows: 1)Brown yellow solid, melting point 74.2–74.6℃; 2)The nuclear magnetic resonance spectrum of the compound is characterized as follows: In deuterated DMSO and deuterated chloroform as solvent, TMS as internal standard, wherein the peaks are assigned as follows: 1 H NMR (400 MHz, DMSO- d 6): δ = 13.00 (s, 1H), 8.02–8.05 (m, 2H), 7.83 (d, J = 2.0 Hz, 1H), 7.77(dd, J =2.0, 8.4 Hz, 1H), 7.61–7.67 (m, 5H), 7.38 (d, J = 8.4 Hz, 1H), 7.23 (dd, J =2.0, 8.8 Hz, 1H), 5.56 (s, 2H), 3.91 (s, 3H). 13 C NMR (100 MHz, CDCl3): δ= 165.8, 163.5, 150.4, 133.3, 130.5, 127.6, 123.9, 120.2, 115.7, 111.3, 56.7. 3) The high resolution mass spectrum of the compound is characterized as: Using electrospray ionization: calcd for C 23 H 18 N4O3 35 Cl + , ([M+H] + ) 433.1067, found433.1065.。

[0041] The physicochemical properties of compound (5) are as follows: 1) Light yellow solid, melting point 182.2-182.9°C; 2) The nuclear magnetic resonance spectrum of the compound is characterized as: In deuterated DMSO as solvent, TMS as internal standard, wherein the peaks are assigned as: 1 H NMR (400 MHz, DMSO- d 6): δ =13.02 (s, 1H), 8.02–8.05 (m, 2H), 7.83 (d, J = 2.0 Hz, 1H), 7.77 (dd, J =2.0, 8.4Hz, 1H), 7.61–7.67 (m, 4H), 7.32–7.38 (m, 2H), 5.76 (s, 1H), 5.56 (s, 2H),3.91 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ = 165.8, 163.5, 150.4, 149.2, 133.3,130.5, 127.6, 125.0, 123.9, 120.2, 115.7, 111.3, 61.4,56.7. 3) The high resolution mass spectrum of the compound is characterized as: Using electrospray ionization: calcd for C 23 H 18 N4O3 79 Br + , ([M+H] + ) 477.0562, found477.0569.。

[0042] The physicochemical properties of compound (6) are as follows: 1) Brownish red solid, melting point 70.5-71.1°C; 2) The nuclear magnetic resonance spectrum characteristics of the compound are as follows: In deuterated DMSO as solvent, TMS as internal standard, wherein the peaks are as follows: 1 H NMR (500 MHz, DMSO- d 6): δ = 13.48 (s, 1H), 8.45 (s, 1H), 8.13 (d, J = 6.8 Hz, 1H), 8.04 (d, J = 6.8 Hz, 2H), 7.62-7.87 (m, 6H), 7.42 (d, J = 6.8 Hz, 1H), 5.58 (s, 2H), 3.92 (s, 3H). 13 C NMR (125 MHz, DMSO- d 6): δ = 165.8, 163.4, 150.4, 149.8, 133.3, 130.5, 127.6, 124.4, 123.9, 120.8, 118.8, 115.6, 111.6, 61.4, 56.7. 3) The high resolution mass spectrum characteristics of the compound are as follows: Using electrospray ionization: calcd for C 23 H 18 N5O5 + , ([M+H] + ) 444.1308, found 444.1310.

[0043] The physicochemical properties of compound (7) are as follows: 1) Brown solid, melting point 221.4-222.1°C; 2) The nuclear magnetic resonance spectrum characteristics of the compound are as follows: In deuterated DMSO as solvent, TMS as internal standard, wherein the peaks are as follows: 1 H NMR (400 MHz, DMSO- d 6): δ = 13.25 (s, 1H), 8.01-8.05 (m, 3H), 7.79-7.86 (m, 3H), 7.61-7.67 (m, 3H), 7.53 (d, J = 8.0 Hz, 1H), 7.41 (d,J = 8.4 Hz, 2H), 5.57 (s, 2H), 3.92 (s, 3H). 13 C NMR (100 MHz, DMSO-) d 6): δ = 165.8, 163.5, 150.5, 149.5, 133.3, 130.5, 127.6, 124.7, 123.9, 120.5, 115.7, 113.0, 111.5, 61.4, 56.7. 3) High-resolution mass spectrometry characteristics of this compound: Electrospray ionization was used: calcd for C 24 H 18 F3N4O3 + , ([M+H] + ) 467.1331, found467.1335.

[0044] The physicochemical properties of compound (8) are as follows: 1) Light green solid, melting point 157.5–158.2℃; 2) The nuclear magnetic resonance (NMR) spectral characteristics of this compound: Using deuterated chloroform and deuterated DMSO as solvents and TMS as an internal standard, the peaks were assigned as follows: 1 H NMR (400 MHz, CDCl3): δ = 8.00–8.04 (m, 4H), 7.58–7.67 (m, 8H), 7.39 (d, J = 8.4 Hz, 1H), 5.59 (s, 2H), 3.91 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ = 168.2, 165.8, 163.3,150.6, 150.1, 133.2, 132.3, 130.4, 128.5, 127.6, 124.8,123.9, 122.0, 115.9,111.6, 61.4, 56.8. 3) High-resolution mass spectrometry characteristics of this compound: Electrospray ionization was used: calcd for C 24 H 18 N4O3SNa + , ([M+Na] +) 465.0997, found465.1003.。

[0045] The physicochemical properties of compound (9) are as follows: 1) light yellow solid, melting point 160.3-160.9℃; 2) the nuclear magnetic resonance spectrum characteristics of the compound are as follows: In deuterated DMSO as solvent, TMS as internal standard, wherein the peak assignment is as follows: 1 H NMR (400 MHz, DMSO- d 6): δ =8.05 (dd, J = 1.2, 8.0 Hz, 2H), 7.79–7.82 (m, 2H),7.58–7.65 (m, 5H), 7.47 (d, J =7.6 Hz, 1H), 7.36–7.41(m, 2H), 5.58 (s, 2H), 3.91 (s, 3H),2.42 (s, 3H). 13 C NMR(100 MHz, DMSO- d 6): δ = 168.3, 168.2, 165.8, 163.3, 150.6, 150.1, 139.9, 133.9,133.2, 132.9, 130.5, 130.3, 128.9, 127.6, 125.7, 124.8, 123.9, 122.0,115.8,111.5, 61.4, 56.8, 21.8. 3) the high resolution mass spectrum characteristics of the compound are as follows: Using electrospray ionization: calcd for C 25 H 21 N4O3S + , ([M+H] + ) 457.1334, found457.1335.。

[0046] The physicochemical properties of compound (10) are as follows: 1) light yellow solid, melting point 175.1-175.6℃; 2) the nuclear magnetic resonance spectrum characteristics of the compound are as follows: In deuterated DMSO and deuterated chloroform as solvent, TMS as internal standard, wherein the peak assignment is as follows: 1 H NMR (400 MHz,DMSO- d 6): δ= 8.03 (s, 2H), 7.91 (d, J = 6.0 Hz, 2H), 7.59–7.63 (m, 5H), 7.38 (s,3H), 5.58 (s, 2H), 3.90 (s, 3H), 2.40 (s, 3H), 13 C NMR (100 MHz, CDCl3): δ =168.0, 167.1, 166.0, 162.0, 161.70, 150.3, 149.2, 142.8, 141.6, 132.1, 129.9, 129.8, 129.1, 127.8, 127.4, 127.2, 127.1, 125.4, 123.4, 121.5, 120.6, 115.1, 110.6, 61.2, 56.2, 21.5. 3) High-resolution mass spectrometry characteristics of this compound: Electrospray ionization was used: calcd for C 25 H 21 N4O3S + , ([M+H] + ) 457.1334, found457.1332.

[0047] The physicochemical properties of compound (11) are as follows: 1) White solid, melting point 87.2–87.6℃; 2) The nuclear magnetic resonance (NMR) spectral characteristics of this compound: Using deuterated DMSO as solvent and TMS as internal standard, the peaks were assigned as follows: 1 H NMR (400 MHz, DMSO- d 6): δ =8.02–8.04 (m, 2H), 7.48–7.67 (m, 8H), 7.39 (d, J = 8.4 Hz, 1H), 7.18 (dd, J =2.0, 8.0 Hz, 1H), 5.58 (s, 2H), 3.91 (s, 3H), 3.87 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ= 168.4, 168.1, 163.3, 160.7, 150.6, 150.1, 133.3, 131.7, 130.5,127.6, 124.7, 123.9, 122.0,121.0, 118.2, 115.8, 113.2, 111.6. 3) High resolution mass spectrum of the compound: Using electrospray ionization: calcd for C 25 H 21 N4O4S + , ([M+H] + ) 473.1285, found 473.1284.

[0048] The physicochemical properties of compound (12) are as follows: 1) White solid, melting point 170.3-170.8°C; 2) The nuclear magnetic resonance spectrum of the compound is characterized: In deuterated DMSO as solvent, TMS as internal standard, wherein each peak is assigned as follows: 1 H NMR (400 MHz, DMSO- d 6): δ = 8.04 (d, J = 6.8 Hz, 2H), 7.94 (d, J = 8.4 Hz, 2H), 7.58-7.67 (m, 7H), 7.38 (d, J = 8.4 Hz, 1H), 5.59 (s, 2H), 3.91 (s, 3H), 1.33 (s, 9H). 13 C NMR (100 MHz, DMSO- d 6): δ = 168.2, 167.9, 165.8, 163.3, 155.2, 150.6, 150.1, 133.3, 130.5, 128.3,127.6, 127.3, 124.8, 123.9, 122.0, 115.8,111.6, 61.4, 56.8, 35.7, 31.8. 3) High resolution mass spectrum of the compound: Using electrospray ionization: calcd for C 28 H 26 N4O3SNa + , ([M+Na] +) 521.1623, found521.1625.。

[0049] The physicochemical properties of compound (13) are as follows: 1) Brown-yellow solid, melting point 172.4-173.2°C; 2) The nuclear magnetic resonance spectrum characteristics of the compound are: In deuterated DMSO as solvent, TMS as internal standard, wherein the peak assignment is: 1 H NMR (400 MHz, DMSO- d 6): δ =8.02–8.04 (m, 3H), 7.57–7.67 (m, 5H), 7.34–7.38 (m, 2H), 6.79–6.80 (m, 1H),5.59 (s, 2H), 3.91 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ = 167.3, 165.8, 163.3,158.2, 150.6, 150.2, 147.3, 145.5, 133.2, 130.5, 127.6,124.4, 123.9, 122.1,115.8, 113.9, 113.2, 111.6, 61.4, 56.8. 3) The high resolution mass spectrum characteristics of the compound are: Using electrospray ionization: calcd for C 22 H 17 N4O4S + , ([M+H] + ) 433.0972, found433.0971.。

[0050] The physicochemical properties of compound (14) are as follows: 1) Light yellow solid, melting point 140.4-141.2°C; 2) The nuclear magnetic resonance spectrum characteristics of the compound are: In deuterated DMSO as solvent, TMS as internal standard, wherein the peak assignment is: 1 H NMR (400 MHz, DMSO- d 6): δ =8.02–8.04 (m, 2H), 7.88 (dd, J = 0.8, 5.2 Hz, 1H), 7.80(dd, J= 0.8, 3.6 Hz, 1H),7.55–7.67 (m, 5H), 7.38 (d, J = 8.4 Hz, 1H), 7.26–7.28 (m, 1H), 5.58 (s, 2H), 3.90 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ = 167.6, 165.8, 163.3, 162.1, 150.6, 150.1, 133.3, 132.4, 131.7, 131.4, 130.5, 129.6, 127.6, 124.5, 123.9, 122.0, 115.8, 111.5, 61.4, 56.8. 3) High-resolution mass spectrometry characteristics of this compound: Electrospray ionization was used: calcd for C 22 H 17 N4O3S2 + , ([M+H] + ) 449.0742, found449.0738.

[0051] The physicochemical properties of compound (15) are as follows: 1) Pale yellow solid, melting point 68.5–69.2℃; 2) The nuclear magnetic resonance (NMR) spectral characteristics of this compound: Using deuterated DMSO as solvent and TMS as internal standard, the peaks were assigned as follows: 1 H NMR (400 MHz, DMSO- d 6): δ =12.74 (s, 1H), 8.14–8.16 (m, 2H), 7.74 (dd, J = 2.0, 8.4 Hz, 2H), 7.62–7.67 (m,5H),7.43 (d, J = 8.4 Hz, 1H), 7.20–7.22 (m, 2H), 5.42 (s, 2H), 3.92 (s, 3H). 13 CNMR (100 MHz, DMSO- d 6): δ= 164.9, 164.6, 151.4, 150.3, 150.3, 132.9, 130.3,127.6, 124.3, 121.1, 117.4, 114.6, 110.6, 65.3, 56.7. 3) High resolution mass spectrum of the compound is characterized as: Using electrospray ionization: calcd for C 23 H 19 N4O3 + , ([M+H] + ) 399.1457, found 339.1451.

[0052] The physicochemical properties of compound (16) are as follows: 1) Yellow solid, melting point 115.5-116.2°C; 2) The nuclear magnetic resonance spectrum of the compound is characterized as: In deuterated DMSO as solvent, TMS as internal standard, wherein the peaks are assigned as: 1 H NMR (400 MHz, DMSO- d 6): δ =12.75 (s, 1H), 7.93-7.97 (m, 2H), 7.66-7.73 (m, 3H), 7.40-7.51 (m, 4H), 7.19-7.23 (m, 2H), 5.42 (s, 2H), 3.92 (s, 3H), 2.44 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ = 164.9, 164.5, 160.6, 151.4, 150.3, 150.2, 131.6, 125.5, 121.1, 119.9, 118.7, 117.4, 114.7, 112.5, 110.7, 65.3, 56.7, 56.4. 3) High resolution mass spectrum of the compound is characterized as: Using electrospray ionization: calcd for C 24 H 21 N4O3 + , ([M+H] + ) 413.1614, found 413.1618.

[0053] The physicochemical properties of compound (17) are as follows: 1) White solid, melting point 190.6-191.2 °C; 2) The nuclear magnetic resonance spectrum characteristics of the compound: In deuterated chloroform and deuterated DMSO as solvent, TMS as internal standard, wherein the peak attribution is: 1 H NMR (400 MHz, CDCl3): δ = 12.74 (s, 1H), 7.71-7.74 (m, 2H), 7.67 (d, J = 12.8 Hz, 2H), 7.52-7.56 (m, 3H), 7.42 (d, J = 8.4 Hz, 1H), 7.22 (d, J = 6.0 Hz, 3H), 5.43 (s, 2H),3.92 (s, 3H), 3.88 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ = 164.9, 164.5, 160.6,151.4, 150.3, 150.2, 131.6, 125.5, 121.1, 119.9, 118.7, 117.4, 114.7, 112.5,110.7, 65.3, 56.7, 56.4. 3) The high resolution mass spectrum characteristics of the compound: Using electrospray ionization: calcd for C 24 H 21 N4O4 + , ([M+H] + ) 429.1563, found429.1567.。

[0054] The physicochemical properties of compound (18) are as follows: 1) White solid, melting point 97.7-98.2 °C; 2) The nuclear magnetic resonance spectrum characteristics of the compound: In deuterated DMSO as solvent, TMS as internal standard, wherein the peak attribution is: 1 H NMR (400 MHz, DMSO- d 6): δ =12.74 (s, 1H), 8.07 (d, J = 8.8 Hz, 2H), 7.72 (dd, J= 2.0, 8.4 Hz, 1H), 7.63–7.66 (m, 4H), 7.53 (d, J = 7.6 Hz, 1H), 7.42 (d, J = 8.4 Hz, 1H), 7.17–7.25 (m,2H), 5.42 (s, 2H), 3.92 (s, 3H), 1.34 (s, 9H). 13 C NMR (100 MHz, DMSO- d 6): δ =164.7, 155.8, 151.4, 150.3, 150.3, 143.8, 135.4, 127.4, 127.1, 123.6, 122.4, 121.6, 121.0, 120.0, 117.5, 114.6, 112.5, 110.6, 65.3, 56.7. 3) High-resolution mass spectrometry characteristics of this compound: Electrospray ionization was used: calcd for C 27 H 26 N4O3Na + , ([M+Na] + ) 477.1902, found477.1903.

[0055] The physicochemical properties of compound (19) are as follows: 1) White solid, melting point 151.4–152.1℃; 2) The nuclear magnetic resonance (NMR) spectral characteristics of this compound: Using deuterated DMSO as solvent and TMS as internal standard, the peaks were assigned as follows: 1 H NMR (400 MHz, DMSO- d 6): δ =12.74 (s, 1H), 8.09 (d, J = 1.2 Hz, 1H), 7.66 (dd, J = 2.0, 8.4 Hz, 2H), 7.61 (d, J = 2.0 Hz, 1H), 7.53 (d, J = 7.6 Hz, 1H), 7.46 (d, J = 3.6 Hz, 1H), 7.42 (d, J=8.4 Hz, 1H), 7.17–7.25 (m, 2H), 6.83–6.84 (m, 1H), 5.42 (s, 2H), 3.91 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ = 164.1, 157.6, 151.5, 150.3, 150.2, 147.9, 143.7, 139.6, 135.3, 123.6, 123.4, 121.0, 120.0, 117.0, 115.5, 113.6, 112.5, 110.6, 65.3, 56.7. 3) High-resolution mass spectrometry characteristics of this compound: Electrospray ionization was used: calcd for C 21 H 17 N4O4 + , ([M+H] + ) 389.1250, found389.1247.

[0056] The physicochemical properties of compound (20) are as follows: 1) White solid, melting point 186.2–186.9℃; 2) The nuclear magnetic resonance (NMR) spectral characteristics of this compound: Using deuterated DMSO as solvent and TMS as internal standard, the peaks were assigned as follows: 1 H NMR (400 MHz, DMSO- d 6): δ =12.75 (s, 1H), 7.96–7.97 (m, 2H), 7.62–7.68 (m, 3H), 7.53 (d, J = 7.6 Hz, 1H), 7.42 (d, J = 8.4 Hz, 1H), 7.32–7.34 (m, 1H), 7.18–7.25 (m, 2H), 5.42 (s, 2H), 3.92 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ= 164.3, 161.0, 151.4, 150.3, 150.2,143.8, 135.4, 132.5, 131.4, 129.7, 125.3, 123.6, 122.5, 121.0, 120.0, 117.2,114.6, 112.5,110.6, 65.3, 56.7. 3) High resolution mass spectrum of the compound: Electrospray ionization: calcd for C 21 H 16 N4O3SNa + , ([M+Na] + ) 427.0841, found427.0844.。

[0057] The physicochemical properties of compound (21) are as follows: 1) White solid, melting point 106.2-106.7°C; 2) The nuclear magnetic resonance spectrum of the compound is characterized: In deuterated DMSO as solvent, TMS as internal standard, wherein the peaks are assigned as follows: 1 H NMR (400 MHz, DMSO- d 6): δ =12.73 (s, 1H), 8.21 (s, 2H), 7.42–7.72 (m, 7H), 7.21(s, 2H), 5.42 (s, 2H),3.92 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ = 151.4, 150.3, 130.3, 130.3, 123.6,121.1, 117.7, 117.5, 117.4, 114.6, 110.7, 65.3, 56.7. 3) High resolution mass spectrum of the compound: Electrospray ionization: calcd for C 23 H 17 FN4O3Na + , ([M+Na] + ) 439.1182, found439.1186.。

[0058] The physicochemical properties of compound (22) are as follows: 1) White solid, melting point 107.6-108.3°C; 2) The compound's nuclear magnetic resonance spectrum characteristics: In deuterated DMSO as solvent, TMS as internal standard, wherein the peak is attributed to: 1 H NMR (400 MHz, DMSO- d 6): δ =12.74 (s, 1H), 8.14 (s, 1H), 7.64–7.74 (m, 7H),7.42 (s, 1H), 7.21 (s, 2H),5.42 (s, 2H), 3.91 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6): δ = 165.2, 162.8, 151.6,150.3, 150.2, 134.1, 132.8, 132.3, 132.1, 128.8, 123.5,121.2, 117.2, 114.7,110.6, 65.3, 56.7. 3) The compound's high resolution mass spectrum characteristics: Using electrospray ionization: calcd for C 23 H 17 N4O3 35 ClNa + , ([M+Na] + ) 455.0887, found455.0890.。

[0059] The compound (23) physical and chemical properties are as follows: 1) White solid, melting point 220.2-220.7℃; 2) The compound's nuclear magnetic resonance spectrum characteristics: In deuterated DMSO as solvent, TMS as internal standard, wherein the peak is attributed to: 1 H NMR (400 MHz, DMSO- d 6): δ =12.75 (s, 1H), 8.09 (d, J = 8.0 Hz, 2H), 7.86 (d, J = 8.0 Hz, 2H), 7.58–7.74 (m,4H), 7.42 (d, J = 8.4 Hz, 1H), 7.21 (s, 2H), 5.42 (s, 2H), 3.92 (s, 3H). 13 C NMR(100 MHz, DMSO-d 6): δ = 151.5, 150.3, 150.2, 133.4, 129.5, 123.6, 121.2, 110.7,65.3, 56.7. 3)The high resolution mass spectrum of the compound is characterized as follows: Using electrospray ionization: calcd for C 23 H 17 N4O3 79 BrNa + , ([M+Na] + ) 499.0382, found499.0386.。

[0060] The physicochemical properties of compound (24) are as follows: 1)White solid, melting point 102.8–103.5℃; 2)The nuclear magnetic resonance spectrum of the compound is characterized as follows: In deuterated chloroform as solvent, TMS as internal standard, wherein the peaks are assigned as follows: 1 H NMR (400 MHz, CDCl3): δ =11.01 (s, 1H), 8.26 (s, 1H), 7.75–7.80(m, 3H), 7.35–7.39 (m, 3H), 7.30 (d, J =7.4 Hz, 1H). 7.23–7.26 (m, 2H), 6.66–6.71 (m, 2H), 6.62 (s, 1H), 6.39 (s,1H), 5.45 (s, 2H),4.12 (d, J = 5.8 Hz, 2H), 2.83 (s, 3H). 13 C NMR (100 MHz,CDCl3): δ =171.2, 170.7, 151.2, 150.4, 149.2, 146.5, 134.6, 132.3, 128.8,128.0, 126.0, 122.7, 120.6, 112.4, 110.8, 100.8,65.4, 54.4, 49.5. 3)The high resolution mass spectrum of the compound is characterized as follows: Using electrospray ionization: calcd for C 25 H 22 N4O2SNa + , ([M+Na] +) 465.1361, found465.1363.。

[0061] The physicochemical properties of compound (25) are as follows: 1) Brown-yellow solid, melting point 130.3-131.1°C; 2) The nuclear magnetic resonance spectrum characteristics of the compound are: In deuterated chloroform as solvent, TMS as internal standard, wherein the peak assignment is: 1 H NMR (400 MHz, CDCl3): δ = 8.25 (s, 1H), 8.14 (dd, J = 1.6, 7.6 Hz, 1H), 7.57 (s, 1H), 7.21–7.25 (m, 3H),7.08 (s, 1H), 7.00–7.04 (m, 1H), 6.94 (d, J = 8.0 Hz, 1H), 6.65–6.70 (m, 2H),6.39 (s, 1H), 5.44 (s, 2H), 4.08 (s, 2H), 3.86 (s, 3H), 2.81 (s, 3H). 13 C NMR(100 MHz, CDCl3): δ = 169.1, 156.9, 150.5, 149.1, 146.6, 146.4, 132.6, 129.6,128.6, 123.3, 122.6, 120.8, 120.6, 111.9, 111.1, 110.9, 105.7, 65.3,55.3,49.5. 3) The high resolution mass spectrum characteristics of the compound are: Using electrospray ionization: calcd for C 26 H 25 N4O3S + , ([M+H] + ) 473.1648, found473.1647.。

[0062] The physicochemical properties of compound (26) are as follows: 1) Brown-yellow solid, melting point 85.8-86.3°C; 2) The nuclear magnetic resonance spectrum characteristics of the compound are: In deuterated chloroform as solvent, TMS as internal standard, wherein the peak assignment is: 1 H NMR (400 MHz, CDCl3): δ=8.36 (s, 1H), 7.69 (d, J = 8.0 Hz, 2H). 7.57 (s, 2H), 7.23–7.26 (m, 2H), 7.19(d, J = 8.0 Hz, 2H), 6.68 (s, 2H), 6.56 (s, 1H), 6.36–6.40 (m, 1H), 5.46 (s,2H), 4.11 (s, 2H), 2.74–2.83 (m, 3H), 2.34 (s, 3H). 13 C NMR (100 MHz, CDCl3): δ =171.0, 151.1, 150.4, 149.0, 146.4, 137.9, 132.3, 131.8, 129.5, 125.8, 122.7, 120.6, 111.6, 110.7, 99.8, 65.1, 54.0, 49.5. 3) High-resolution mass spectrometry characteristics of this compound: Electrospray ionization was used: calcd for C 26 H 24 N4O2SNa + , ([M+Na] + ) 479.1518, found479.1519.

[0063] The physicochemical properties of compound (27) are as follows: 1) Pale yellow solid, melting point 80.3–80.9℃; 2) The nuclear magnetic resonance (NMR) spectral characteristics of this compound: Using deuterated chloroform as solvent and TMS as internal standard, the peaks were assigned as follows: 1 H NMR (400 MHz, CDCl3): δ =8.30 (s, 1H), 7.73 (d, J = 8.8 Hz, 2H). 7.57 (s, 2H), 7.23–7.25 (m, 2H), 6.91(d, J = 8.8 Hz, 2H), 6.67–6.72 (m, 2H), 6.48 (s, 1H), 6.41 (s, 1H), 5.45 (s, 2H), 4.11 (s, 2H), 3.81 (s, 3H), 2.85 (s, 3H). 13 C NMR (100 MHz, CDCl3):δ = 170.9, 159.5, 151.0, 150.4, 149.1, 146.4, 132.4, 127.6, 127.2, 122.7, 120.6, 114.1, 112.0, 110.8, 98.9, 65.3, 55.3, 49.5. 3) The high resolution mass spectrum characteristics of the compound: Using electrospray ionization: calcd for C 26 H 25 N4O3S + , ([M+H] + ) 473.1647, found 473.1656.

[0064] The physicochemical properties of compound (28) are as follows: 1) Brown-yellow solid, melting point 98.5-99.2°C; 2) The nuclear magnetic resonance spectrum characteristics of the compound: In deuterated chloroform as solvent, TMS as internal standard, wherein the peak assignment is as follows: 1 H NMR (400 MHz, CDCl3): δ = 8.08 (s, 1H), 7.74-7.78 (m, 2H), 7.57 (s, 2H), 7.23-7.26 (m, 2H), 7.03-7.08 (m, 2H), 6.67-6.74 (m, 2H), 6.55 (s, 1H), 6.40 (d, J = 1.6 Hz, 1H), 5.45 (s, 2H), 4.13 (d, 2H), 2.90 (s, 3H). 13 C NMR (100 MHz, CDCl3): δ = 170.7, 163.8, 161.3, 150.4, 150.3, 149.2, 146.6, 132.2, 131.0, 130.9, 127.7, 127.6, 122.7, 120.6, 115.6, 112.6, 110.6, 100.4, 65.4, 54.5, 49.5. 3) The high resolution mass spectrum characteristics of the compound: Using electrospray ionization: calcd for C 25 H 22 FN4O2S + , ([M+H] +) 461.1447, found 461.1448.

[0065] The physicochemical properties of compound (29) are as follows: 1) pink solid, melting point 100.6-101.3°C; 2) the nuclear magnetic resonance spectrum characteristics of the compound are as follows: The peaks are assigned as follows with deuterated chloroform as solvent and TMS as internal standard: 1 H NMR (400 MHz, CDCl3): δ = 7.98 (s, 1H), 7.73 (d, J = 8.8 Hz, 2H), 7.57 (s, 2H), 7.34 (d, J = 8.4 Hz, 2H),7.22-7.26 (m, 2H),6.68-6.76 (m, 2H), 6.62 (s, 1H), 6.43 (d, J = 1.6 Hz, 1H),5.45 (s, 2H), 4.15 (s, 2H), 2.96 (s, 3H). 13 C NMR (100 MHz, CDCl3): δ = 170.7,150.4, 150.1, 149.2, 146.5, 133.6, 133.1, 132.1, 128.9, 127.1, 122.7,120.6,112.6, 110.7, 101.2, 65.4, 54.4, 49.5. 3) the high resolution mass spectrum characteristics of the compound are as follows: Using electrospray ionization: calcd for C 25 H 22 N4O2S 35 Cl + , ([M+H] + ) 477.1152, found 477.1156.

[0066] The physicochemical properties of compound (30) are as follows: 1) orange-red solid, melting point 98.3-98.8°C; 2) the nuclear magnetic resonance spectrum characteristics of the compound are as follows: The peaks are assigned as follows with deuterated chloroform as solvent and TMS as internal standard: 1 H NMR (400 MHz, CDCl3): δ =7.94-7.96 (m, 2H), 7.71 (d,J = 8.0 Hz, 1H), 7.57 (s, 2H), 7.39–7.41 (m, 1H), 7.21–7.27 (m, 3H), 6.75 (d, J = 8.0 Hz, 1H), 6.71 (dd, J = 1.6, 8.0 Hz, 1H), 6.66(s, 1H), 6.45 (d, J = 1.6 Hz, 1H), 5.45 (s, 2H), 4.20 (d, J = 5.8 Hz (2H), 2.94 (s, 3H). 13 C NMR (100 MHz, CDCl3): δ = 170.7, 150.3, 149.6, 149.1, 146.5, 136.6, 132.1, 130.7, 130.2, 128.9, 124.3, 122.9, 122.7, 120.6, 112.4, 110.7, 101.9, 65.3, 54.4, 49.4. 3) High-resolution mass spectrometry characteristics of this compound: Electrospray ionization was used: calcd for C 25 H 22 N4O2 79 BrS + , ([M+H] + ) 521.0645, found521.0647.

[0067] The physicochemical properties of compound (31) are as follows: 1) White solid, melting point 126.2–126.8℃; 2) The nuclear magnetic resonance (NMR) spectral characteristics of this compound: Using deuterated chloroform as solvent and TMS as internal standard, the peaks were assigned as follows: 1 H NMR (400 MHz, CDCl3): δ =7.81 (s, 1H), 7.67 (d, J = 8.4 Hz, 2H), 7.57 (s, 2H), 7.50 (d, J= 8.4 Hz, 2H), 7.24–7.26 (m, 2H), 6.70–6.77 (m, 2H), 6.64 (s, 2H), 6.45 (s, 1H), 5.46 (s, 2H), 4.18 (s, 2H), 2.99 (s, 3H). 13 C NMR (100 MHz, CDCl3): δ = 170.6, 150.4,150.1, 149.3, 146.6, 133.6, 132.2, 131.8, 127.68, 127.4, 122.7, 121.8, 120.6,112.6, 110.7, 101.3, 65.5, 54.4, 49.5. 3) High-resolution mass spectrometry characteristics of this compound: Electrospray ionization was used: calcd for C 25 H 22 N4O2 79 BrS + , ([M+H] + ) 521.0647, found521.0645.

[0068] The physicochemical properties of compound (32) are as follows: 1) Pink solid, melting point 97.2–97.9℃; 2) The nuclear magnetic resonance (NMR) spectral characteristics of this compound: Using deuterated chloroform as solvent and TMS as internal standard, the peaks were assigned as follows: 1 H NMR (400 MHz, CDCl3): δ =7.90 (d, J = 2.0 Hz, 1H), 7.54–7.60 (m, 3H), 7.42 (d, J = 8.4 Hz, 1H), 7.22–7.26(m, 2H), 6.79 (d, J = 8.0 Hz, 1H), 6.73 (dd, J = 1.6, 8.4 Hz, 1H), 6.67 (s, 1H), 6.52 (d, J = 1.6 Hz, 1H), 5.43 (s, 2H), 4.24 (d, J = 5.8 Hz, 2H), 3.11 (s, 3H). 13 CNMR (100 MHz, CDCl3): δ= 170.5, 150.4, 149.3, 148.9, 146.7, 134.6, 132.9,131.6, 130.7, 127.7, 125.0, 122.7, 120.6, 113.1, 110.9, 102.3, 65.6, 54.7,49.5. 3) High resolution mass spectrum of the compound: Using electrospray ionization: calcd for C 25 H 21 N4O2S 35 Cl2 + , ([M+H] + ) 511.0762, found511.0764.。

[0069] The physicochemical properties of compound (33) are as follows: 1) Yellow solid, melting point 96.4-96.9°C; 2) The nuclear magnetic resonance spectrum of the compound is characterized: In deuterated chloroform as solvent, TMS as internal standard, wherein each peak is assigned as follows: 1 H NMR (400 MHz, CDCl3): δ =7.64–7.68 (m, 2H), 7.51–7.55 (m, 3H), 7.43 (dd, J = 1.2, 7.6 Hz, 1H), 7.21–7.26(m, 3H), 6.82–6.84 (m, 1H). 6.74 (s, 2H), 6.74 (s, 2H), 6.59 (s, 1H), 6.45(s, 1H), 5.40 (s, 2H), 4.25(s, 2H), 3.51 (s, 3H). 13 C NMR (100 MHz, CDCl3): Cytospora mandshurica =169.4, 150.5, 149.5, 149.2, 146.6, 146.6, 132.2, 131.9, 130.6, 129.1, 128.6,123.7, 122.7, 120.4, 113.9, 111.3, 104.6, 65.7,55.3, 49.3. 3) High resolution mass spectrum of the compound: Using electrospray ionization: calcd for C 25 H 21 N5O4SNa + , ([M+Na]+ ) 510.1213, found510.1212.。

[0070] Bioactivity determination test of vanillin compounds spliced with polycyclic heterocycle 1. Test pathogenic bacteria: Valsa mali (Coker) Wagg (1) Fusarium solani ), Rhizoctonia solani Kuhn (2) Fusarium graminearum ), Fusarium graminearum Schwabe (3) Pyricularia ), Pyricularia grisea Sacc (4) grisea Botrytis cinerea ), Botrytis cinerea Pers (5) Alternaria solani ) and Alternaria solani (Jones) R. H. Jones (6) Fusarium oxysporum ) and Fusarium oxysporum f. sp. niveum (6) ​ ).

[0071] 2. Test sample and reagent: The sample is the above-prepared vanillin compounds spliced with polycyclic heterocycle (1)-(33).

[0072] The solvent is acetone.

[0073] 3. Bioassay method: (1) Mycelial growth rate method The inhibition effect of the above-prepared vanillin compounds spliced with polycyclic heterocycle (1)-(33) on the mycelial growth of Valsa mali (Coker) Wagg, Rhizoctonia solani Kuhn, Fusarium graminearum Schwabe, Pyricularia grisea Sacc, Botrytis cinerea Pers and Alternaria solani (Jones) R. H. Jones is determined by mycelial growth rate: after all the test samples are dissolved with acetone, they are mixed with melted PDA medium, poured into sterile culture dishes, about 15 mL per dish, to prepare drug-containing medium with a concentration of 100 μg / mL (the content of acetone in the drug-containing medium does not exceed 0.5%, v / v). After the medium cools and solidifies, the test strains are activated in advance and punched into 4 mm-diameter fungus cakes along the edge of the colony, inoculated in the center of the drug-containing medium, and each drug is repeated 3 times. At the same time, a blank control test without drug is set. Cultivate at a suitable temperature until the colony diameter of the blank control group reaches 2 / 3 of the diameter of the culture dish. The colony diameters of each treatment group and the control group are measured by cross method, and the mycelial growth inhibition rate of each drug treatment group is calculated according to formula-1, and the corresponding standard deviation is calculated. The results of the antibacterial bioassay are shown in Table-1.

[0074] Mycelial growth inhibition rate = (average colony diameter of control group-average colony diameter of treatment group) / (average colony diameter of control group-4 mm) x 100% (formula-1) Table-1 Antifungal activity of vanillin compounds spliced with polycyclic heterocycle on the mycelial growth of six pathogenic fungi As shown in Table-1, the antibacterial bioassay results show that almost all the compounds (except compound 12) prepared in the present application have an inhibitory effect on the growth of Valsa mali mycelium that is higher than that of vanillin; a total of 23 compounds (except compounds 3, 8, 14, 19, 24, 26, 29, 30 and 32) have an inhibitory effect on the growth of Rhizoctonia bataticola mycelium that is higher than that of vanillin; a total of 25 compounds (except compounds 5, 7, 8, 12, 18, 19, 26 and 28) have an inhibitory effect on the growth of Gibberella zeae mycelium that is higher than that of vanillin; compounds 1, 2, 4, 5, 16, 18, 21, 30 and 33 have an inhibitory effect on the growth of Pyricularia grisea mycelium that is higher than that of vanillin; and all the compounds tested have an inhibitory effect on the growth of Alternaria solani mycelium that is higher than that of vanillin.

[0075] As can be found above, the inhibitory activities of most of the prepared derivatives on Valsa mali, Rhizoctonia bataticola and Gibberella zeae are higher than that of vanillin, indicating that the introduction of benzimidazole, 1, 3, 4-thiadiazole, thiazole or 1, 3, 4-oxadiazole heterocyclic structure into the molecular skeleton of vanillin helps to improve the antibacterial activity of such substances. Derivative 10 shows excellent inhibitory activity on Valsa mali, with an inhibition rate of 100%, which is much higher than that of vanillin, indicating that the derivative has high inhibitory activity on fungal mycelium and has great application potential.

[0076] (2) Activity determination by spore germination method The above prepared multi-heterocycle spliced vanillin compounds (1)-(33) were tested for inhibiting spore germination activity of four plant pathogenic fungi, Fusarium oxysporum, Rhizoctonia bataticola, Botrytis cinerea and Alternaria solani, by spore germination method: a. Preparation of spore suspension: the pathogenic fungi were cultured on PDA medium at 25 ± 1°C for 2 weeks, then the colonies were gently scraped off with a scraper, washed with sterile water, filtered with double-layer gauze to remove mycelium and medium, washed repeatedly with sterile water for 3 times, and finally diluted with sterile water to the required spore concentration (100 times, about 20-40 spores per field under a microscope); b. Preparation of drug: the sample to be tested was dissolved in a certain amount of DMSO, diluted with 0.1% Tween-80 sterile water solution to prepare a 1 mg / mL stock solution, and then 5-7 series of mass concentrations were set according to the activity of the drug, and the final content of organic solvent was not more than 2% (v / v); c. Drug treatment: the prepared sample solution was mixed with the spore suspension in equal volume to obtain the required final concentration. 2% DMSO and 0.1% Tween-80 sterile water solution were used as the control group. The above mixture was dropped on a concave glass slide with a micro-sampler, placed in a culture dish with a shallow layer of water, covered and placed in a suitable temperature incubator for culture. The spore germination on the glass slide was observed under a microscope. When the germ tube after spore germination was greater than the short diameter of the spore, it was considered to be germinated. When the spore germination rate of the blank control was greater than 90%, the spore germination rate of each treatment group was observed and calculated (formula-2), and the corrected spore germination inhibition rate of each treatment group was calculated by formula-3, and the inhibition concentration of each compound, 95% confidence interval, etc. were calculated. The biological assay results are shown in Table-2.

[0077] Spore germination rate = germinated spore number / total number of spores counted × 100% (formula-2); Corrected spore germination inhibition rate = (average spore germination rate of control group - average spore germination rate of treatment group) / average spore germination rate of control group × 100% (formula-3).

[0078] Table-2 Inhibitory germination effect of multi-heterocycle spliced vanillin compounds on spores of four plant pathogenic fungi As shown in Table-2: the antibacterial biological assay results show that the prepared multi-heterocycle spliced vanillin compounds (benzimidazole vanillin, thiazole vanillin, thiazole vanillin, and oxadiazole vanillin derivatives) in the application can treat four kinds of pathogenic bacterial spores, and the inhibitory concentration of raw vanillin on the spores of four tested fungi (except for watermelon fusarium wilt fungus) is larger, and the inhibitory activity is general. The inhibitory germination activity of compound 13 on the spores of red kidney bean root rot fungus is better than that of vanillin. The inhibitory germination activity of derivatives 1, 19 and 29 on the spores of tomato gray mold fungus is the most outstanding, and the inhibitory concentration is 7.2, 17.3 and 8.0 μg / mL respectively, and the inhibitory activity is far more than that of vanillin, and it has the potential to be used as a potential tomato gray mold fungus spore germination inhibitor.

[0079] 4. Conclusion In summary, the multi-heterocycle spliced vanillin compounds (benzimidazole vanillin, thiazole vanillin, thiazole vanillin, and oxadiazole vanillin derivatives) in the application can be used for preparing high-efficiency new antibacterial agents, and applied to the prevention and control of apple tree rot fungus, red kidney bean root rot fungus, wheat scab fungus, millet fusarium wilt fungus, tomato gray mold fungus, tomato early blight fungus and watermelon fusarium wilt fungus.

[0080] The specification of the application is considered to be illustrative rather than limiting, and on the basis of the application, those skilled in the art can make some substitutions and modifications to some technical features according to the disclosed technical content without creative labor, and all are within the protection scope of the application.

Claims

1. A vanillin compound spliced with multiple heterocycles, characterized in that, The multi-heterocycle is one of a benzimidazole ring, a thiadiazole ring, an oxadiazole ring, and a thiazole ring; the vanillin compound spliced with the multi-heterocycle is one of vanillin derivatives containing a benzimidazole ring, a thiadiazole ring, an oxadiazole ring, and a thiazole ring, and the chemical general formula of the vanillin compound spliced with the multi-heterocycle is shown in the following formula: The chemical formula is: The compounds (1)-(7) are benzimidazole vanillin derivatives, (8)-(14) are thiazole vanillin derivatives, (15)-(24) are oxadiazole vanillin derivatives, and (25)-(33) are thiazole vanillin derivatives.

2. The polyheterocyclic spliced vanillin compound according to claim 1, characterized in that, The thiazole ring is a 1,3,4-thiazole ring, and the oxadiazole ring is a 1,3,4-oxadiazole ring.

3. The process for preparing a polyheterocyclic spliced vanillin compound according to claim 1 or 2, wherein The method comprises the following steps: condensation reaction and condensation-cyclization reaction.

4. The process for the preparation of polyheterocyclic spliced vanillin derivatives as claimed in claim 3, wherein, The condensation reaction comprises A1 or A2, and the condensation-cyclization reaction comprises one of B1, B2, B3, and B4. A1: 0.8 mmol of 2-(chloromethyl)-5-phenyl-1,3,4-oxadiazole and 1.0 mmol of vanillin are dissolved in 2 mL of acetone, 1.2 mmol of potassium carbonate and 0.1 mmol of potassium iodide are added, and the reaction is stirred at 55 ℃, the reaction progress is tracked and detected, after the reaction is completed, the filtrate is concentrated under reduced pressure, and compound 3-methoxy-4-[ (5-phenyl-1,3,4-oxadiazol-2-yl) methoxy] benzaldehyde is obtained by silica gel thin layer chromatography separation; A2: 1.4 mmol of vanillin, 1.1 mmol of 2-(chloromethyl)-1H-benzimidazole, 1.7 mmol of potassium carbonate and 0.1 mmol of potassium iodide are dissolved in 2 mL of acetone; the reaction is stirred at 55 ℃, the reaction progress is tracked and detected, after the reaction is completed, the filtrate is concentrated under reduced pressure, and compound 2-[ (3-methoxy-4-formylphenoxy) methyl]-1H-benzimidazole is obtained by silica gel thin layer chromatography separation; B1: 0.4 mmol of 3-methoxy-4-[ (5-phenyl-1,3,4-oxadiazol-2-yl) methoxy] benzaldehyde, 0.4 mmol of substituted o-phenylenediamine and 0.4 mmol of sodium metabisulfite are dissolved in 2 mL of dimethyl sulfoxide, the reaction is stirred at 140 ℃, the reaction progress is tracked and detected, after the reaction is completed, the reaction solution is cooled to room temperature, 10 mL of water is added to quench the reaction, 40 mL of ethyl acetate is used to extract 5 times, the organic phase is combined, anhydrous sodium sulfate is used to dry the organic phase, filtration is performed, and the filtrate is concentrated under reduced pressure, followed by silica gel thin layer chromatography separation; B2: 0.4 mmol of substituted aromatic hydrazine and 0.4 mmol of 3-methoxy-4-[ (5-phenyl-1,3,4-oxadiazol-2-yl) methoxy] benzaldehyde are dissolved in 2 mL of methanol, the reaction is stirred at 65 ℃, the reaction progress is tracked and detected, after the reaction is completed, the reaction solution is concentrated under reduced pressure, 2 mL of toluene is added to dissolve the residue at room temperature, 0.5 mmol of Lawesson's reagent and 0.5 mmol of 4- (dimethylamino) pyridine are sequentially added to the mixture, the reaction is stirred at 110 ℃, the reaction progress is tracked and detected, after the reaction is completed, the reaction solution is concentrated under reduced pressure, and the residue is separated by silica gel thin layer chromatography. B3: 0.6 mmol of substituted aromatic hydrazide and 0.4 mmol of compound 2-[(3-methoxy-4-formylphenoxy)methyl]-1H-benzimidazole were dissolved in 2 mL of methanol, and the reaction was stirred at 65°C. The reaction progress was tracked, and after the reaction was completed, the reaction mixture was concentrated under reduced pressure. 2 mL of dichloromethane-methanol was added to the residue, and 0.5 mmol of diethyl iodo phenyl ether was added to the mixture. The reaction was stirred at room temperature. The reaction progress was tracked, and after the reaction was completed, the reaction mixture was concentrated under reduced pressure. The residue was separated by silica gel thin layer chromatography; B4: 0.5 mmol of compound 2-[(3-methoxy-4-formylphenoxy)methyl]-1H-benzimidazole and 0.8 mmol of substituted 4-phenyl-thiazol-2-amine were dissolved in 2 mL of toluene, and the reaction was stirred at 110°C. The reaction progress was tracked, and after the reaction was completed, the reaction mixture was concentrated under reduced pressure. 2 mL of anhydrous ethanol was added to the residue, and 3.2 mmol of sodium borohydride was added to the mixture in batches. The reaction was stirred at room temperature. The reaction progress was tracked, and after the reaction was completed, the reaction mixture was concentrated under reduced pressure. The pH value was adjusted to 7 with saturated sodium bicarbonate solution, and the organic phase was extracted with 40 mL of ethyl acetate three times. The organic phase was combined and dried over anhydrous sodium sulfate. The reaction mixture was filtered, concentrated under reduced pressure, and separated by silica gel thin layer chromatography.

5. The process for the preparation of polyheterocyclic spliced vanillin derivatives as claimed in claim 4, wherein, The compounds (1)-(7) are obtained by condensation reaction A1 and condensation-cyclization reaction B1; the compounds (8)-(14) are obtained by condensation reaction A1 and condensation-cyclization reaction B2; the compounds (15)-(24) are obtained by condensation reaction A2 and condensation-cyclization reaction B3; and the compounds (25)-(33) are obtained by condensation reaction A2 and condensation-cyclization reaction B4.

6. The process for the preparation of polyheterocyclic spliced vanillin derivatives as claimed in claim 5, wherein, The substituted o-phenylenediamine is one of 4-methyl-o-phenylenediamine, 3,4-dimethyl-o-phenylenediamine, 4-chloro-o-phenylenediamine, 4-bromo-o-phenylenediamine, 4-nitro-o-phenylenediamine, or 4-trifluoromethyl-o-phenylenediamine; The substituted aromatic hydrazide is one of 3-methyl aromatic hydrazide, 4-methyl aromatic hydrazide, 3-methoxy aromatic hydrazide, 4-tert-butyl aromatic hydrazide, 2-furan aromatic hydrazide, 2-thiophene aromatic hydrazide, 4-fluoro aromatic hydrazide, 2-chloro aromatic hydrazide, or 4-bromo aromatic hydrazide; The substituted benzhydrazide is one of 2-methoxy benzhydrazide, 4-methyl benzhydrazide, 4-methoxy benzhydrazide, 4-fluoro benzhydrazide, 4-chloro benzhydrazide, 3-bromo benzhydrazide, 4-bromo benzhydrazide, 3,4-dichloro benzhydrazide, or 2-nitro benzhydrazide.

7. The application of the polyheterocycle-spliced vanillin compound of claim 1 or 2 or the polyheterocycle-spliced vanillin compound obtained by the preparation method of any one of claims 3-6.

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