Triazole-cis-alkenyl amide compound as well as preparation method and anti-inflammatory application thereof

Abstract

The invention discloses a triazole-cis-alkenyl amide compound as well as a preparation method and anti-inflammatory application thereof, and relates to the technical field of biological medicines. The triazole-cis-alkenyl amide compound provided by the invention has an excellent effect in the aspects of resisting inflammation and treating allergic rhinitis diseases.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a triazole-cis-enamide compound and a preparation method and anti-inflammatory application thereof. Background Art

[0002] Allergic rhinitis (AR), also known as allergic rhinitis, is a non-infectious inflammatory disease caused by activation of immunoglobulin (IgE) after inhalation of environmental allergens. Its main clinical symptoms are paroxysmal sneezing, runny nose, nasal congestion, nasal itching, and also accompanied by symptoms such as eye itching and tearing. According to statistics, about 10%-40% of the world's population is currently affected by allergic rhinitis, and the trend is increasing year by year, becoming a global health issue of concern.

[0003] Lansiumamide B is a cis-enamide compound isolated from the kernel of Phellodendron chinense. It has multiple pharmacological activities such as treating obesity, lowering blood lipids, anti-diabetes, anti-snake venom, and anti-inflammatory. Its structural characteristics are cinnamamide part and cis-enamide part. 1,2,4-Triazole is a stable azole compound derived from the substitution of nitrogen atom for the 4-carbon atom of pyrazole. It often exists in two tautomeric forms. Due to its dipole effect, hydrogen bond formation ability, rigidity and solubility, 1,2,4-triazole can produce high affinity with biological receptors and become an important pharmacophore. In addition, 1,2,4-triazole can also be used as a bioisostere of amides, esters and carboxylic acids. Therefore, 1,2,4-triazole is a dominant skeleton in the field of drug research and development and is commonly found in various clinical drugs, including anticancer drugs, antiviral drugs, anticonvulsant drugs, antifungal drugs, antianxiety drugs, hypnotics, skeletal muscle relaxants, antimigraine drugs, antiplatelet drugs, antidepressants and aromatase inhibitors.

[0004] The present invention aims to develop anti-inflammatory drugs for treating allergic rhinitis by combining the cis-enamide structure with the 1,2,4-triazole structure commonly used in medicinal chemistry through structural transition. Summary of the invention

[0005] The purpose of the present invention is to provide a triazole-cis-enamide compound and a preparation method and anti-inflammatory application thereof, so as to solve the problems existing in the above-mentioned prior art.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] The present invention provides a 1,2,4-triazole-cis-enamide compound and a pharmaceutically acceptable salt or solvate thereof, the structural formula of which is shown in formula (I):

[0008]

[0009] In formula (I), R is hydrogen, C6-C14 aryl, five- to fourteen-membered heteroaryl, C1-C10 cycloalkyl, five- to fourteen-membered heterocyclyl, C1-C6 straight-chain alkyl or C1-C6 branched-chain alkyl;

[0010] The C6-14 aryl, five- to fourteen-membered heteroaryl, C6-10 cycloalkyl or five- to fourteen-membered heterocyclic group is substituted by one or more substituents selected from halogen, halogenated C1-C6 alkyl, carbonyl, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, carboxyl, cyano, hydroxyl, carbonyl, phenyl-SO2-, nitro, C1-C6 alkyloxycarbonyl; and the five- to fourteen-membered heteroaryl or five- to fourteen-membered heterocyclic group contains 1-3 heteroatoms selected from N, O and S.

[0011] Furthermore, the halogen is F, Cl, Br or I.

[0012] Furthermore, the 1,2,4-triazole-cis-enamide compound and its pharmaceutically acceptable salt or solvate are selected from any one of the following:

[0013]

[0014] Furthermore, the pharmaceutically acceptable salt is a pharmaceutically acceptable salt, including an inorganic acid salt, an organic acid salt, an alkyl sulfonate or an aryl sulfonate.

[0015] Specifically, the inorganic acid salt includes but is not limited to hydrochloride, hydrobromide, hydroiodide, isothiosulfate, nitrate, sulfate, bisulfate, persulfate, phosphate, and bicarbonate;

[0016] Specifically, the organic acid salt includes but is not limited to formates, acetates, propionates, butyrates, valerates, pivalates, camphorates, adipates, alginate, aspartates, benzoates, maleates, fumarates, lactates, nicotinates, succinates, tartrates, citrates, digluconates, glycerophosphates, cysteine ​​salts, oxalates, palmitates, pectinates, 3-phenylpropionates, thiocyanates, glutamate, and oleates;

[0017] Specifically, the alkyl sulfonate includes but is not limited to methyl sulfonate, ethyl sulfonate, and camphor sulfonate;

[0018] Specifically, the aryl sulfonate includes but is not limited to benzene sulfonate, p-toluene sulfonate, and 2-naphthalene sulfonate.

[0019] Furthermore, the solvate includes a solvate of the 1,2,4-triazole-cis-enamide compound and water, ethanol, isopropanol, ether, ethylene glycol, acetone or glycerol.

[0020] The present invention also provides a method for preparing the above-mentioned 1,2,4-triazole-cis-enamide compounds and their pharmaceutically acceptable salts or solvates, comprising the following steps:

[0021]

[0022] a) dissolving the compounds of formula I and formula II in acetonitrile solution in a stoichiometric ratio of 1:1, and reacting at room temperature for 1-3 hours;

[0023] b) dissolving the product compound of formula III obtained in step a) in dry dichloromethane, and dropping the mixture into a dry dichloromethane solution of NBS-dimethyl sulfide (3 equivalents) cooled to -78°C, reacting at -78°C for 0.5-2h, then warming to room temperature (20-25°C) and reacting for 2-5h;

[0024] c) dissolving the compounds of formula V and formula VI in water in a stoichiometric ratio of 1:1.2, adding sodium carbonate in small amounts several times, and reacting at room temperature for 2-5 hours;

[0025] d) dissolving the product compound of formula IV obtained in step b) and the product compound of formula VII obtained in step c) in dry toluene at a stoichiometric ratio of 1:1.2, adding triethylamine, and reacting at 110° C.-130° C. for 1-3 h;

[0026] e) dissolving the product compound of formula VIII obtained in step d) in an ammonia methanol solution and reacting at room temperature for 2-8 days;

[0027] f) reacting the product compound of formula IX obtained in step e) with the compound of formula X at a stoichiometric ratio of 1.2:1 at 40° C.-80° C. for 0.5-3 h;

[0028] g) The product compound of formula XI obtained in step f) is reacted with sodium hydride (1.6 equivalents) at -20°C-5°C with stirring for 0.5-1h, and then iodomethane (1.6 equivalents) is added at this temperature, and finally the temperature is raised to 20°C-50°C and stirred for reaction for 2-6h to obtain a compound of formula XII, which is the structural compound represented by formula (I).

[0029] wherein R is hydrogen, C6-C14 aryl, five- to fourteen-membered heteroaryl, C1-C10 cycloalkyl, five- to fourteen-membered heterocyclyl, C1-C6 straight-chain alkyl, or C1-C6 branched-chain alkyl;

[0030] The C6-14 aryl, five- to fourteen-membered heteroaryl, C6-10 cycloalkyl or five- to fourteen-membered heterocyclic group is substituted by one or more substituents selected from halogen, halogenated C1-C6 alkyl, carbonyl, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, carboxyl, cyano, hydroxyl, carbonyl, phenyl-SO2-, nitro, C1-C6 alkyloxycarbonyl; and the five- to fourteen-membered heteroaryl or five- to fourteen-membered heterocyclic group contains 1-3 heteroatoms selected from N, O and S.

[0031] The present invention further provides the use of the above-mentioned 1,2,4-triazole-cis-enamide compounds and their pharmaceutically acceptable salts or solvates in the preparation of anti-inflammatory drugs.

[0032] The present invention further provides the use of the above-mentioned 1,2,4-triazole-cis-enamide compounds and their pharmaceutically acceptable salts or solvates in the preparation of drugs for treating allergic rhinitis.

[0033] The present invention discloses the following technical effects:

[0034] The invention provides a triazole-cis-enamide compound, which has excellent effects in anti-inflammatory and treating allergic rhinitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0036] Figure 1 It shows the inhibitory effect of 1,2,4-triazole-cis-enamide compounds on the mRNA levels of IL-6 and IL-1β inflammatory factors in Raw264.7 cells in effect verification example 1. Figure A shows the IL-6 RNA expression level of Raw264.7 cells stimulated by LPS after being treated with different compounds, and Figure B shows the IL-1β m expression level of Raw264.7 cells stimulated by LPS after being treated with different compounds; Figures C and E show the IL-6, IL-1β, and TNF-α mRNA expression levels of Raw264.7 cells stimulated by LPS after being treated with different concentrations of 2125 compounds (compared with the Control group). ### P<0.001; compared with LPS group * P<0.05, ** P<0.01, *** P<0.001).

[0037] Figure 2The compound in effect verification example 2 relieves the symptoms of rhinitis in AR mouse model (compared with the Control group). #### P<0.0001; compared with LPS group ** P<0.01, *** P < 0.001, **** P<0.0001).

[0038] Figure 3 The total number of inflammatory cells in the nasal lavage fluid of mice in the effect verification example 2 (compared with the control group ## P<0.01; compared with AR group * P<0.05, ** P<0.01).

[0039] Figure 4 Gimsa staining of nasal lavage cells in mice in effect verification example 2 (A) and the number of eosinophils (B) and mast cells (C) (compared with the Control group #### P<0.0001; compared with AR group *** P < 0.001, **** P<0.0001).

[0040] Figure 5 Expression of HIS in mouse serum (compared with the Control group) #### P<0.0001; compared with AR group * P<0.05, *** P < 0.001, **** P<0.0001).

[0041] Figure 6 The figure shows the pathological picture of the mouse lung tissue stained with HE in effect verification example 2 (magnification: 400×; Scale bar=100 μm).

[0042] Figure 7 The figure shows the PAS staining of mouse lung tissue in effect verification example 2 (magnification: 400×; Scale bar=100 μm). DETAILED DESCRIPTION

[0043] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0044] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0045] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0046] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0047] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0048] The room temperature described in the embodiment of the present invention refers to 20-25°C.

[0049] Example 1 Synthesis of intermediate IV of the present invention:

[0050]

[0051] Step 1: Take a 100mL egg-shaped bottle, weigh phenylhydrazine I (1.08g, 10mmol) in the bottle and add a magnetic bar, add acetonitrile (15mL) to dilute and stir, weigh ethyl glyoxylate II (2.04g, 10mmol, 50wt% in toluene) in a sample bottle, dilute with acetonitrile (5mL) and slowly drip into the egg-shaped bottle, then wash the sample bottle with acetonitrile (10mL) and pour into the egg-shaped bottle, stir and react for 2h at room temperature, wait for the reaction to be complete, remove the solvent with a rotary evaporator, separate and purify by silica gel chromatography column, use petroleum ether as eluent, and obtain yellow solid III (1.88g, 9.8mmol), yield: 98%.

[0052] Step 2: Take a 100mL two-necked flask, a vacuum head and a magnet and place them in an oven for drying. Take out the two-necked flask and assemble it, then evacuate it. After the temperature of the flask cools to room temperature, replace it with argon. Weigh N-bromosuccinimide (2.67g, 15mmol) and add it to the two-necked flask and add dry anhydrous dichloromethane (20mL). Under 0℃ ice bath conditions, slowly drip dimethyl sulfide (2.25mL, 30mmol) into the bottle and react for 15min. Lower the temperature to -78℃, weigh III (961mg, 5mmol) in the sample bottle, add dry anhydrous dichloromethane (10mL) to dissolve it and drip it into the two-necked flask. Then wash the sample bottle with dry anhydrous dichloromethane (6mL) twice and pour it into the two-necked flask. Keep the reaction at -78℃ for 1h, then let it heat up naturally and stir for 3h. After the reaction, water (15 mL) was added dropwise to the two-necked flask to quench the reaction, and dichloromethane (3×15 mL) was used for extraction. The organic phase was washed with water (3×15 mL), then washed once with a saturated sodium chloride solution (20 mL), once with a saturated sodium sulfite solution (20 mL), and finally washed once with water (20 mL). The organic phase was dried over anhydrous sodium sulfate, and the solvent was removed from the dried organic phase by a rotary evaporator. The product was separated and purified by a silica gel chromatography column using petroleum ether as an eluent to obtain a yellow solid IV (1.19 g, 4.4 mmol) with a yield of 88%.

[0053] Example 2 Synthesis of the final product 2121 of the present invention

[0054] (1) Synthesis of Compound 2

[0055]

[0056] Take a 25mL egg-shaped bottle, weigh benzaldehyde (531mg, 5mmol) in the bottle, add a magnetic bar and water (5mL), then add hydroxylamine hydrochloride (417mg, 6mmol), add sodium carbonate (1.06g, 10mmol) in small amounts and several times under room temperature stirring, and react at room temperature for 3h. After the reaction is complete, extract with ether (3×10mL), then wash once with saturated sodium chloride solution (10mL), collect the organic phase and dry it with anhydrous sodium sulfate, remove the solvent from the dried organic phase with a rotary evaporator, and obtain a white solid that is directly used in the next step.

[0057] (2) Synthesis of Compound 3

[0058]

[0059] Take 100mL sealed tube, weigh 2 (363mg, 3mmol), IV (678mg, 2.5mmol) respectively, pour into the sealed tube and add a magnetic bar, draw dry anhydrous toluene (15mL) into the sealed tube, fill the sealed tube with argon (for 1min), draw dry triethylamine (0.69mL, 5mmol) and drip into the sealed tube, tighten the sealed tube stopper, place the sealed tube in a 120℃ oil bath, heat and stir to react for 2h. Remove the oil bath, let the temperature drop to room temperature, remove toluene by rotary evaporation, add water (20mL), extract with dichloromethane (3×20mL), wash once with saturated sodium bicarbonate solution (20mL), wash once with saturated sodium chloride solution (20mL), collect the organic phase and dry with anhydrous sodium sulfate, remove the solvent with a rotary evaporator after drying, separate and purify by silica gel chromatography column, use petroleum ether as eluent, and obtain 403mg (compound 3) of yellow oily liquid, yield: 55%.

[0060] (3) Synthesis of Compound 4

[0061]

[0062] Take 15mL sealed tube, weigh compound 3 (403mg, 1.37mmol) and add a magnetic bar, inject ammonia methanol solution (2.74mL), fill the sealed tube with argon (for 1min), tighten the stopper, and stir to react for 3 days. After the reaction is complete, collect the organic phase, remove the solvent with a rotary evaporator, and then separate and purify it through a silica gel chromatography column with petroleum ether and acetone (V / V=1:1) as eluent to obtain 330mg of white solid (compound 4), yield: 90%.

[0063] (4) Synthesis of Compound 5

[0064]

[0065] Take 10mL The Schlenk tube and the magnetic bar were dried, and vacuum was applied after assembly. When the temperature dropped to room temperature, argon gas was introduced, and compound 4 (291 mg, 1.1 mmol), cesium carbonate (407 mg, 1.25 mmol), and cuprous iodide (9.5 mg, 0.05 mmol) were weighed in. Dry tetrahydrofuran (5 mL) was taken to dilute (Z)-2-iodovinylbenzene (243 mg, 1 mmol) and then injected into the Schlenk tube. DMEDA (11 μL, 0.1 mmol) was then added dropwise. The Schlenk tube was sealed and placed in an oil bath at 60°C for heating and stirring to react for 3 h. When the reaction was completed, the oil bath was removed and the temperature was naturally dropped to room temperature. The reaction solution was filtered through diatomaceous earth, and the diatomaceous earth was rinsed with ethyl acetate (3×20 mL). The organic phase was collected and the solvent was removed by rotary evaporator. After separation and purification on a silica gel chromatography column, petroleum ether and ethyl acetate (V / V=3:1) were used as eluents to obtain 304 mg of a yellow solid (compound 5) with a yield of 83%.

[0066] (5) Synthesis of Compound 2121

[0067]

[0068] Take a 25mL two-necked flask, a vacuum head and a magnetic dryer, assemble and evacuate, wait until the temperature drops to room temperature, replace with argon, weigh compound 5 (275mg, 0.75mmol), draw dry DMF (3mL) into the two-necked flask, add 60% pure sodium hydride (45mg, 1.2mmol) under 0℃ ice bath conditions, keep 0℃ stirring reaction for 30min, then slowly add iodomethane (68μL, 1.2mmol), slowly return to room temperature and stir the reaction for 3h. After the reaction is completed, add water under ice bath conditions to quench the reaction, extract with ether (3×20mL), wash with saturated sodium chloride solution (20mL), dry the organic phase with anhydrous sodium sulfate, and then use a rotary evaporator to remove the solvent, separate and purify by silica gel chromatography column, use petroleum ether and ethyl acetate (V / V=5:1) as eluents, and obtain 228mg of yellow solid (compound 2121), yield: 80%.

[0069] The obtained compound 2121 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were:

[0070] Main conformational isomers: 1 H NMR (400MHz, CDCl3) δ: 7.53-7.31 (m, 14H), 7.26-7.21 (m, 1H), 6.94 (d, J = 8.8Hz, 1H), 6.06 (d, J = 8.8Hz, 1H), 3.16 (s, 3H). 13C NMR (100MHz, CDCl3) δ: 161.9, 157.2, 154.2, 137.8, 135.1, 130.4, 130.3, 129.5 (2×C), 129.3, 129.2 (2×C), 128.8 (2×C), 128.6 (2×C), 128.5 (2×C), 127.7, 127.3, 125.5 (2×C), 121.2, 35.8. Minor conformational isomers: 1 H NMR (400MHz, CDCl3) δ: 7.53-7.31 (m, 14H), 7.26-7.21 (m, 1H), 7.06 (d, J = 9.2Hz, 1H), 6.22 (d, J = 9.2Hz, 1H), 3.31 (s, 3H). 13 C NMR (100 MHz, CDCl3) δ: 162.5, 157.0, 154.5, 137.8, 135.5, 130.6, 130.4, 129.6 (2×C), 129.3, 129.0 (2×C), 128.7 (2×C), 128.6 (2×C), 128.4 (2×C), 127.5, 127.1, 125.5 (2×C), 121.1, 38.7. HRMS (ESI) m / z calculated for C 24 H 21 ON4 + [M+H] + 381.1710, the measured value is 381.1716.

[0071] By choosing different aldehydes, compounds 2122-2142 can be obtained:

[0072] Example 3 Synthesis of Compound 2122:

[0073] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 4-fluorobenzaldehyde to obtain compound 2122 with a yield of 80%. The main conformational isomers were: 1 H NMR(400MHz, CDCl3)δ:7.54-7.40(m,5H),7.38-7.30(m,6H),7.26-7.21(m,1H) ,7.04-7.00(m,2H),6.91(d,J=8.8Hz,1H),6.05(d,J=8.8Hz,1H),3.15(s,3H). 13CNMR (100MHz, CDCl3) δ: 163.9 (d, J = 250Hz), 161.7, 157.1, 153.4, 137.7, 135.0, 131.3 (d, J = 8.5Hz, 2×C), 130.2, 129.6 ( 2×C),129.5,128.8(2×C),128.6(2×C),127.7,125.5(2×C),123.4(d,J=3.3Hz),121.4,115.9(d,J=21.9Hz,2×C),35.8. 19 F NMR (376MHz, CDCl3) δ: -109.0. Minor conformational isomers: 1 H NMR(400MHz, CDCl3)δ:7.54-7.40(m,5H),7.38-7.30(m,6H),7.26-7.21(m,1H) ,7.04-7.00(m,2H),7.05(d,J=9.2Hz,1H),6.22(d,J=9.2Hz,1H),3.30(s,3H). 13 C NMR (100MHz, CDCl3) δ: 163.9 (d, J = 250Hz), 162.3, 157.0, 153.6, 137.7, 135.5, 131.3 (d, J = 8.5Hz, 2×C), 130.2, 129.7 ( 2×C), 129.3, 129.0 (2×C), 128.4 (2×C), 127.5, 125.5 (2×C), 123.3 (d, J=3.3Hz), 120.3, 116.0 (d, J=22.0Hz, 2×C), 38.7. 19 F NMR (376 MHz, CDCl3) δ: -109.0. HRMS (ESI) m / z calculated value is C 24 H 20 ON4F + [M+H] + 399.1616, the measured value is 399.1611.

[0074] Example 4 Synthesis of Compound 2123:

[0075] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was isobutylaldehyde to obtain compound 2123 with a yield of 85%. The main conformational isomers were: 1H NMR(400MHz, CDCl3)δ:7.52-7.45(m,3H),7.44-7.35(m,2H),7.31-7.25(m,4H),7.22-7.18(m,1H),6.8 6(d,J=8.8Hz,1H),5.98(d,J=8.8Hz,1H),3.11(sept,J=6.8Hz,1H),3.10(s,3H),1.29(d,J=6.8Hz,6H). 13 C NMR (100MHz, CDCl3) δ: 162.1, 161.4, 156.5, 137.0, 135.1, 130.3, 129.5 (2×C), 129.4, 128.7 (2×C), 128.5 (2×C), 127.5, 125.5 (2×C), 120.7, 35.7, 25.9, 21.6 (2×C). Minor conformational isomers: 1 H NMR(400MHz, CDCl3)δ:7.52-7.45(m,3H),7.44-7.35(m,2H),7.31-7.25(m,4H),7.22-7.18(m,1H),7.0 2(d,J=9.2Hz,1H),6.17(d,J=9.2Hz,1H),3.22(s,3H),3.11(sept,J=6.8Hz,1H),1.31(d,J=6.8Hz,6H). 13 C NMR (100 MHz, CDCl3) δ: 162.7, 161.7, 156.4, 137.0, 135.5, 130.3, 129.6 (2×C), 129.4, 128.9 (2×C), 128.3 (2×C), 127.3, 125.5 (2×C), 119.8, 38.5, 25.9, 21.6 (2×C). HRMS (ESI) m / z calculated value is C 21 H 23 ON4 + [M+H] + 347.1866, the measured value is 347.1868.

[0076] Example 5 Synthesis of Compound 2124:

[0077] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 2-thiophenecarboxaldehyde to obtain compound 2124 with a yield of 85%. The main conformational isomers were: 1H NMR(400MHz, CDCl3)δ:7.56-7.42(m,5H),7.40-7.37(m,1H),7.34-7.29(m,4H),7.25-7.20(m,1H),7.08 (d,J=3.6Hz,1H),6.94(dd,J=4.8,4.0Hz,1H),6.91(d,J=8.8Hz,1H),6.04(d,J=8.8Hz,1H),3.13(s,3H). 13 C NMR (100MHz, CDCl3) δ: 161.6, 157.0, 149.9, 137.3, 135.0, 130.3, 130.2, 129.8, 129.7 (2×C), 129.6, 129.5, 128.8 (2×C), 128.5 (2×C), 127.7, 127.6, 126.7 (2×C), 121.2, 35.8. Minor conformational isomers: 1 H NMR(400MHz, CDCl3)δ:7.56-7.42(m,5H),7.40-7.37(m,1H),7.34-7.29(m,4H),7.25-7.20(m,1H),7.10 (d,J=3.2Hz,1H),7.04(d,J=9.2Hz,1H),6.94(dd,J=4.8,4.0Hz,1H),6.21(d,J=9.2Hz,1H),3.28(s,3H). 13 C NMR (100 MHz, CDCl3) δ: 162.2, 156.8, 150.1, 137.3, 135.5, 130.5, 130.2, 129.8, 129.7 (2×C), 129.6, 129.3, 129.0 (2×C), 128.4 (2×C), 127.8, 127.5, 126.7 (2×C), 120.2, 38.6. HRMS (ESI) m / z calculated value is C 22 H 19 ON4S + [M+H] + 387.1274, the measured value is 387.1272.

[0078] Example 6 Synthesis of Compound 2125:

[0079] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was cyclohexylcarboxaldehyde to obtain compound 2125 with a yield of 88%. The main conformational isomers were: 1H NMR(400MHz, CDCl3)δ:7.52-7.45(m,3H),7.43-7.35(m,2H),7.32-7.26(m,4H),7.24-7.19(m,1H),6.86(d, J=8.8Hz,1H),5.98(d,J=8.8Hz,1H),3.11(s,3H),2.78-2.72(m,1H),1.81-1.65(m,7H),1.33-1.16(m,3H). 13 C NMR (100MHz, CDCl3) δ: 162.1, 160.6, 156.6, 137.1, 135.2, 130.4, 129.5 (2×C), 129.4, 128.7 (2×C), 128.5 (2×C), 127.5, 125.5 (2×C), 120.7, 35.8, 35.4, 31.7 (2×C), 25.9 (2×C), 25.5. Minor conformational isomers: 1 H NMR(400MHz, CDCl3)δ:7.52-7.45(m,3H),7.43-7.35(m,2H),7.32-7.26(m,4H),7.24-7.19(m,1H),7.02(d, J=9.2Hz,1H),6.18(d,J=9.2Hz,1H),3.22(s,3H),2.78-2.72(m,1H),1.81-1.65(m,7H),1.33-1.16(m,3H). 13 C NMR (100 MHz, CDCl3) δ: 162.8, 160.8, 156.5, 137.1, 135.6, 130.4, 129.6 (2×C), 129.4, 129.0 (2×C), 128.3 (2×C), 127.4, 125.5 (2×C), 119.8, 38.6, 35.4, 31.7 (2×C), 25.9 (2×C), 25.5. HRMS (ESI) m / z calculated for C 24 H 27 ON4 + [M+H] + 387.2174, the measured value is 387.2182.

[0080] Example 7 Synthesis of Compound 2126:

[0081] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 4-bromobenzaldehyde to obtain compound 2126 with a yield of 85%. The main conformational isomers were: 1H NMR (400MHz, CDCl3) δ: 7.52-7.29 (m, 13H), 7.25-7.21 (m, 1H), 6.89 (d, J = 8.8Hz, 1H), 6.05 (d, J = 8.8Hz, 1H), 3.15 (s, 3H). 13 C NMR (100MHz, CDCl3)δ:161.6,157.2,153.3,137.6,135.0,131.9(2×C),130.6(2×C),130.2,129.7(2×C),129.6,128.7(2×C),128.6(2×C),127.7,126.1,125.5(2×C),125.1,121.5,35.8.Minor conformational isomers: 1 H NMR (400MHz, CDCl3) δ: 7.52-7.29 (m, 13H), 7.25-7.21 (m, 1H), 7.05 (d, J = 9.2Hz, 1H), 6.22 (d, J = 9.2Hz, 1H), 3.30 (s, 3H). 13 C NMR (100 MHz, CDCl3) δ: 162.3, 157.1, 153.5, 137.6, 135.5, 132.0 (2×C), 130.6 (2×C), 130.2, 129.8 (2×C), 129.3, 129.0 (2×C), 128.4 (2×C), 127.5, 126.0, 125.5 (2×C), 125.3, 120.3, 38.7. HRMS (ESI) m / z calculated for C 24 H 20 ON + [M+H] + 459.0815, measured value is 459.0807.

[0082] Example 8 Synthesis of Compound 2127:

[0083] The synthesis steps were the same as those in Example 2. Formaldehyde was selected as the aldehyde reagent in the first step to obtain compound 2127 with a yield of 80%. The main conformational isomers were: 1 H NMR (400MHz, CDCl3) δ: 8.56 (s, 1H), 7.67 (d, J = 8.0Hz, 2H), 7.57-7.49 (m, 2H), 7.46-7.40 (m, 1H), 7.36-7.31(m,4H),7.28-7.22(m,1H),6.81(d,J=8.8Hz,1H),6.07(d,J=8.8Hz,1H),3.15(s,3H). 13C NMR (100MHz, CDCl3) δ: 161.5, 158.4, 141.0, 136.7, 135.0, 130.2, 129.9 (2×C), 128.8, 128.7 (2×C), 128.6 (2×C), 127.8, 121.6, 120.3 (2×C), 35.9. Minor conformational isomers: 1 H NMR (400MHz, CDCl3) δ: 8.61 (s, 1H), 7.72 (d, J = 8.0Hz, 2H), 7.57-7.49 (m, 2H), 7.46-7.40 (m, 1H), 7.36-7.31(m,4H),7.28-7.22(m,1H),7.05(d,J=9.2Hz,1H),6.23(d,J=9.2Hz,1H),3.26(s,3H). 13 C NMR (100 MHz, CDCl3) δ: 161.2, 158.2, 141.2, 136.7, 135.0, 130.2, 130.0 (2×C), 129.0 (2×C), 128.9, 128.5 (2×C), 127.6, 121.6, 120.3 (2×C), 38.7. HRMS (ESI) m / z calculated value is C 18 H 17 N4O + [M+H] + 305.1397, the measured value is 305.1401.

[0084] Example 9 Synthesis of Compound 2128:

[0085] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was propionaldehyde to obtain compound 2128 with a yield of 89%. The main conformational isomers were: 1 H NMR(400MHz, CDCl3)δ:7.52-7.39(m,5H),7.32-7.27(m,4H),7.24-7.19(m,1H),6.89(d,J=8 .8Hz,1H),6.00(d,J=8.8Hz,1H),3.11(s,3H),2.82(q,J=7.6Hz,2H),1.31(t,J=7.6Hz,3H). 13 C NMR (100MHz, CDCl3) δ: 162.0, 157.6, 156.6, 137.0, 135.1, 130.4, 129.5 (2×C), 129.3, 128.8 (2×C), 128.5 (2×C), 127.6, 125.0 (2×C), 120.7, 35.8, 20.2, 12.1. Minor conformational isomers: 1H NMR(400MHz, CDCl3)δ:7.52-7.39(m,5H),7.32-7.27(m,4H),7.24-7.19(m,1H),7.03(d,J=9 .2Hz,1H),6.18(d,J=9.2Hz,1H),3.24(s,3H),2.84(q,J=7.6Hz,2H),1.29(t,J=7.6Hz,3H). 13 C NMR (100 MHz, CDCl3) δ: 162.6, 157.8, 156.4, 136.9, 135.5, 130.4, 129.6 (2×C), 129.4, 129.0 (2×C), 128.3 (2×C), 127.4, 125.0 (2×C), 119.9, 38.6, 20.2, 12.1. HRMS (ESI) m / z calculated value is C 20 H 21 ON4 + [M+H] + 333.1710, the measured value is 333.1712.

[0086] Example 10 Synthesis of Compound 2129:

[0087] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 1-naphthaldehyde to obtain compound 2129 with a yield of 83%. The main conformational isomers were: 1 H NMR (400MHz, CDCl3) δ: 7.93 (d, J = 7.2Hz, 1H), 7.87 (d, J = 8.0Hz, 1H), 7.83 (d, J = 8.0Hz, 1H), 7. 52-7.31(m,8H),7.26-7.15(m,6H),6.95(d,J=8.8Hz,1H),6.11(d,J=8.8Hz,1H),3.21(s,3H). 13 C NMR (100 MHz, CDCl3) δ: 161.8, 157.3, 153.1, 137.3, 134.9, 133.6, 131.3, 131.0, 130.2, 129.2, 129.1 (2×C), 128.7 (2×C), 128.6 (2×C), 128.5, 128.4, 127.7, 127.3, 126.6, 125.2, 125.1, 124.9, 124.1 (2×C), 122.0, 35.7. Minor conformational isomers: 1H NMR (400MHz, CDCl3) δ: 7.93 (d, J = 7.2Hz, 1H), 7.87 (d, J = 8.0Hz, 1H), 7.83 (d, J = 8.0Hz, 1H), 7. 52-7.31(m,8H),7.26-7.15(m,6H),7.10(d,J=9.2Hz,1H),6.24(d,J=9.2Hz,1H),3.38(s,3H). 13 C NMR (100 MHz, CDCl3) δ: 162.4, 157.2, 153.4, 137.3, 135.5, 133.6, 131.3, 131.1, 130.2, 129.3, 129.0 (2×C), 128.7 (2×C), 128.6 (2×C), 128.5, 128.3, 127.5, 127.4, 126.7, 125.2, 125.1, 125.0, 124.1 (2×C), 120.2, 38.7. HRMS (ESI) m / z calculated for C 28 H 23 ON4 + [M+H] + 431.1866, the measured value is 431.1850.

[0088] Example 11 Synthesis of Compound 2130:

[0089] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 4-methylbenzaldehyde to obtain compound 2130 with a yield of 80%. The main conformational isomers were: 1 H NMR(400MHz, CDCl3)δ:7.47-7.30(m,11H),7.26-7.21(m,1H),7.13(d,J=8.0H z,2H),6.94(d,J=8.8Hz,1H),6.05(d,J=8.8Hz,1H),3.15(s,3H),2.35(s,3H). 13 C NMR (100MHz, CDCl3) δ: 162.0, 157.1, 154.4, 140.7, 138.0, 135.1, 130.4, 129.4 (2×C), 129.3 (2×C), 129.2, 129.0 (2×C), 128.8 (2×C), 128.6 (2×C), 127.6, 125.5 (2×C), 124.4, 121.1, 35.8, 21.5. Minor conformational isomers: 1H NMR(400MHz, CDCl3)δ:7.47-7.30(m,11H),7.26-7.21(m,1H),7.13(d,J=8.0H z,2H),7.06(d,J=9.2Hz,1H),6.22(d,J=8.8Hz,1H),3.31(s,3H),2.36(s,3H). 13 C NMR (100 MHz, CDCl3) δ: 162.6, 156.9, 154.6, 140.9, 138.0, 135.5, 130.5, 129.6 (2×C), 129.5 (2×C), 129.1, 129.0 (2×C), 128.8 (2×C), 128.4 (2×C), 127.4, 125.8 (2×C), 124.2, 120.1, 38.7, 21.5. HRMS (ESI) m / z calculated for C 25 H 23 ON4 + [M+H] + 395.1866, the measured value is 395.1860.

[0090] Example 12 Synthesis of Compound 2131:

[0091] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 3,3-dimethylbutyraldehyde to obtain compound 2131 with a yield of 87%. The main conformational isomers were: 1 H NMR (400MHz, CDCl3)δ:7.53-7.27(m,9H),7.25-7.20(m,1H),6.84(d,J=8.8Hz,1H),6.01(d,J=8.8Hz,1H),3.13(s,3H),2.72(s,2H),0.87(s,9H). 13 C NMR (100MHz, CDCl3) δ: 162.1, 156.6, 154.9, 137.4, 135.0, 130.4, 129.4 (2×C), 128.8 (2×C), 128.6 (2×C), 127.7, 126.3, 126.2 (2×C), 121.3, 39.0, 35.7, 32.7, 29.6 (3×C). Minor conformational isomers: 1 H NMR (400MHz, CDCl3) δ: 7.53-7.27(m,9H),7.25-7.20(m,1H),7.04(d,J=9.2Hz,1H),6.19(d,J=9.2Hz,1H),3.25(s,3H),2.76(s,2H),0.91(s,9H). 13C NMR (100 MHz, CDCl3) δ: 162.7, 156.5, 155.2, 136.9, 135.6, 130.7, 129.6 (2×C), 129.0 (2×C), 128.4 (2×C), 127.4, 126.3, 126.2 (2×C), 119.8, 38.7, 35.7, 32.7, 29.7 (3×C). HRMS (ESI) m / z calculated value is C 23 H 27 ON4 + [M+H] + 375.2179, the measured value is 375.2189.

[0092] Example 13 Synthesis of Compound 2132:

[0093] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was n-butyraldehyde to obtain compound 2132 with a yield of 87%. The main conformational isomers were: 1 H NMR(400MHz, CDCl3)δ:7.53-7.37(m,5H),7.32-7.27(m,4H),7.23-7.19(m,1H),6.87(d,J=8.8Hz,1H) ,5.99(d,J=8.8Hz,1H),3.11(s,3H),2.75(t,J=7.6Hz,2H),1.81-1.71(m,2H),0.91(t,J=7.6Hz,3H). 13 C NMR (100MHz, CDCl3) δ: 162.0, 156.6, 156.5, 137.0, 135.1, 130.4, 129.5 (2×C), 129.3, 128.7 (2×C), 128.5 (2×C), 127.6, 125.2 (2×C), 120.8, 35.8, 28.4, 21.2, 13.8. Minor conformational isomers: 1 H NMR(400MHz, CDCl3)δ:7.53-7.37(m,5H),7.32-7.27(m,4H),7.23-7.19(m,1H),7.03(d,J=9.2Hz,1H) ,6.18(d,J=8.8Hz,1H),3.23(s,3H),2.75(t,J=7.6Hz,2H),1.81-1.71(m,2H),0.91(t,J=7.6Hz,3H). 13C NMR (100 MHz, CDCl3) δ: 162.6, 156.7, 156.5, 137.0, 135.5, 130.4, 129.6 (2×C), 129.3, 129.0 (2×C), 128.3 (2×C), 127.4, 125.2 (2×C), 119.9, 38.6, 28.4, 21.2, 13.8. HRMS (ESI) m / z calculated for C 21 H 23 ON4 + [M+H] + 347.1866, the measured value is 347.1868.

[0094] Example 14 Synthesis of Compound 2133:

[0095] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was selected as mesitylene-3-benzaldehyde to obtain compound 2133 with a yield of 78%. The main conformational isomers were: 1 H NMR(400MHz, CDCl3)δ:7.38-7.30(m,5H),7.28-7.25(m,3H),7.24-7.19(m,2H),6.87(d, J=8.8Hz,1H),6.86(s,2H),6.05(d,J=8.8Hz,1H),3.18(s,3H),2.30(s,3H),1.94(s,6H). 13 C NMR (100MHz, CDCl3) δ: 162.0, 157.4, 153.4, 140.3, 137.6 (2×C), 137.4, 134.9, 130.4, 129.2 (2×C), 128.8 (2×C), 128.7 (2×C), 128.6 (2×C), 128.3, 127.8, 125.1, 122.6 (2×C), 122.0, 35.7, 21.4, 19.9 (2×C). Minor conformational isomers: 1 H NMR(400MHz, CDCl3)δ:7.38-7.30(m,5H),7.28-7.25(m,3H),7.24-7.19(m,2H),7.08(d, J=9.2Hz,1H),6.86(s,2H),6.22(d,J=9.2Hz,1H),3.30(s,3H),2.30(s,3H),1.98(s,6H). 13C NMR (100 MHz, CDCl3) δ: 161.7, 157.2, 153.6, 140.5, 137.6 (2×C), 137.5, 135.6, 129.4, 129.3 (2×C), 129.1 (2×C), 128.5 (2×C), 128.4 (2×C), 128.3, 127.5, 125.0, 122.6 (2×C), 119.9, 38.8, 21.4, 19.9 (2×C). HRMS (ESI) m / z calculated value is C 27 H 27 ON4 + [M+H] + 423.2179, the measured value is 423.2171.

[0096] Example 15 Synthesis of Compound 2134:

[0097] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 2-methylbenzaldehyde to obtain compound 2134 with a yield of 83%. The main conformational isomers were: 1 H NMR (400MHz, CDCl3) δ: 7.36-7.28 (m, 8H), 7.26-7.16 (m, 6H), 6.90 (d, J = 8.8Hz, 1H), 6.06 (d, J = 8.8Hz, 1H), 3.17 (s, 3H), 2.09 (s, 3H). 13 C NMR (100MHz, CDCl3) δ: 161.8, 157.1, 153.9, 137.6, 137.4, 135.0, 130.8, 130.5, 130.4 (2×C), 130.3, 129.3 (2×C), 128.7 (2×C), 128.6 (2×C), 128.5, 127.7, 126.1, 123.8 (2×C), 121.7, 35.8, 19.9. Minor conformational isomers: 1 HNMR(400MHz, CDCl3)δ:7.36-7.28(m,8H),7.26-7.16(m,6H),7.07(d,J=9.2Hz,1H),6.22(d,J=9.2Hz,1H),3.32(s,3H),2.12(s,3H). 13C NMR (100 MHz, CDCl3) δ: 162.4, 157.0, 154.2, 137.6, 137.4, 135.5, 130.9, 130.6, 130.4 (2×C), 130.3, 129.4, 129.3 (2×C), 129.0 (2×C), 128.4 (2×C), 127.5, 126.2, 123.8 (2×C), 120.1, 38.7, 19.9. HRMS (ESI) m / z calculated for C 25 H 23 ON4 + [M+H] + 395.1866, the measured value is 395.1871.

[0098] Example 16 Synthesis of Compound 2135:

[0099] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 2-furancarboxaldehyde to obtain compound 2135 with a yield of 84%. The main conformational isomers were: 1 H NMR(400MHz, CDCl3)δ:7.53-7.41(m,6H),7.34-7.28(m,4H),7.25-7.21(m,1H),6.90(d,J=8.8 Hz,1H),6.66(d,J=3.6Hz,1H),6.43(dd,J=3.6,1.6Hz,1H),6.04(d,J=8.8Hz,1H),3.14(s,3H). 13 C NMR (100 MHz, CDCl3) δ: 161.6, 157.3, 146.4, 144.7, 141.9, 137.6, 135.0, 130.1, 129.9, 129.3 (2×C), 128.8 (2×C), 128.6 (2×C), 127.7, 126.0 (2×C), 121.5, 113.9, 111.8, 35.8. Minor conformational isomers: 1 H NMR(400MHz, CDCl3)δ:7.53-7.41(m,6H),7.34-7.28(m,4H),7.25-7.21(m,1H),7.03(d,J=9.2 Hz,1H),6.66(d,J=3.6Hz,1H),6.43(dd,J=3.6,1.6Hz,1H),6.22(d,J=9.2Hz,1H),3.26(s,3H). 13C NMR (100 MHz, CDCl3) δ: 162.2, 157.2, 146.7, 144.9, 141.8, 137.6, 135.5, 130.0, 129.5 (2×C), 129.2, 129.0 (2×C), 128.4 (2×C), 127.5, 126.1 (2×C), 120.2, 114.0, 111.8, 38.6. HRMS (ESI) m / z calculated for C 22 H 19 O2N4 + [M+H] + 371.1503, the measured value is 371.1496.

[0100] Example 17 Synthesis of Compound 2136:

[0101] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 2-naphthaldehyde to obtain compound 2136 with a yield of 82%. The main conformational isomers were: 1 H NMR (400MHz, CDCl3) δ: 8.08 (s, 1H), 7.82 (d, J = 8.0Hz, 1H), 7.77 (d, J = 8.4Hz, 1H), 7.76 (d, J = 7.6Hz, 1 H),7.55-7.31(s,12H),7.27-7.22(m,1H),6.97(d,J=8.8Hz,1H),6.08(d,J=8.8Hz,1H),3.18(s,3H). 13 C NMR (100 MHz, CDCl3) δ: 161.9, 157.3, 154.3, 137.9, 135.1, 133.8, 132.8, 130.3, 129.7, 129.5 (2×C), 129.4, 128.8 (2×C), 128.7 (2×C), 128.6, 128.3, 127.8, 127.7, 127.6, 126.9, 125.5 (2×C), 125.4, 124.5, 121.3, 35.8. Minor conformational isomers: 1 H NMR (400MHz, CDCl3) δ: 8.14 (s, 1H), 7.82 (d, J = 8.0Hz, 1H), 7.77 (d, J = 8.4Hz, 1H), 7.76 (d, J = 7.6Hz, 1 H),7.55-7.31(s,12H),7.27-7.22(m,1H),7.09(d,J=9.2Hz,1H),6.24(d,J=9.2Hz,1H),3.35(s,3H). 13C NMR (100 MHz, CDCl3) δ: 162.5, 157.1, 154.5, 137.9, 135.5, 133.8, 132.8, 130.3, 129.6, 129.5 (2×C), 129.4, 129.0 (2×C), 128.7 (2×C), 128.6, 128.4, 127.8, 127.7, 127.5, 126.9, 125.5 (2×C), 125.4, 124.4, 120.2, 38.7. HRMS (ESI) m / z calculated for C 28 H 23 ON4 + [M+H] + 431.1866, the measured value is 431.1875.

[0102] Example 18 Synthesis of Compound 2137:

[0103] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was acetaldehyde to obtain compound 2137 with a yield of 82%. The main conformational isomers were: 1 H NMR(400MHz, CDCl3)δ:7.52-7.43(m,5H),7.33-7.28(m,4H),7.24-7.20(m, 1H), 6.90 (d, J = 8.8Hz, 1H), 6.02 (d, J = 8.8Hz, 1H), 3.11 (s, 3H), 2.54 (s, 3H). 13 C NMR (100MHz, CDCl3) δ: 161.9, 156.6, 152.8, 137.1, 135.1, 130.4, 129.5 (2×C), 129.2, 128.8 (2×C), 128.5 (2×C), 127.6, 124.7 (2×C), 120.7, 35.8, 13.3. Minor conformational isomers: 1 H NMR(400MHz, CDCl3)δ:7.52-7.43(m,5H),7.33-7.28(m,4H),7.24-7.20(m, 1H),7.03(d,J=9.2Hz,1H),6.19(d,J=9.2Hz,1H),3.25(s,3H),2.57(s,3H). 13 C NMR (100 MHz, CDCl3) δ: 162.5, 156.5, 153.0, 137.1, 135.5, 130.4, 129.6 (2×C), 129.4, 129.0 (2×C), 128.4 (2×C), 127.4, 124.7 (2×C), 120.1, 38.6, 13.3. HRMS (ESI) m / z calculated value is C19 H 19 ON4 + [M+H] + 319.1553, the measured value is 319.1546.

[0104] Example 19 Synthesis of Compound 2138:

[0105] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 2-chlorobenzaldehyde to obtain compound 2138 with a yield of 86%. The main conformational isomers were: 1 H NMR (400MHz, CDCl3) δ: 7.48-7.38 (m, 3H), 7.37-7.27 (m, 9H), 7.26-7.20 (m, 2H), 6.88 (d, J = 8.8Hz, 1H), 6.07 (d, J = 8.8Hz, 1H), 3.18 (s, 3H). 13 C NMR (100MHz, CDCl3) δ: 161.6, 157.3, 151.7, 137.4, 135.0, 133.9, 132.1, 132.0, 130.2 (2×C), 129.2 (2×C), 128.8, 128.7 (2×C), 128.6 (2×C), 127.9, 127.7, 127.2, 123.7 (2×C), 121.9, 35.8. Minor conformational isomers: 1 H NMR (400MHz, CDCl3) δ: 7.48-7.38 (m, 3H), 7.37-7.27 (m, 9H), 7.26-7.20 (m, 2H), 7.06 (d, J = 9.2Hz, 1H), 6.23 (d, J = 9.2Hz, 1H), 3.31 (s, 3H). 13 C NMR (100 MHz, CDCl3) δ: 162.2, 157.2, 152.0, 137.4, 135.5, 133.9, 132.1, 132.0, 130.3 (2×C), 129.4 (2×C), 129.3 (2×C), 129.0, 128.4 (2×C), 127.9, 127.7, 127.5, 123.8 (2×C), 120.2, 38.7. HRMS (ESI) m / z calculated for C 24 H 20 ON4Cl + [M+H] + 415.1320, measured value is 415.1327.

[0106] Example 20 Synthesis of Compound 2139:

[0107] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 3-methylbenzaldehyde to obtain compound 2139 with a yield of 85%. The main conformational isomers were: 1 H NMR (400MHz, CDCl3) δ: 7.47-7.31 (m, 10H), 7.25-7.14 (m, 4H), 6.93 (d, J = 8.8Hz, 1H), 6.05 (d, J = 8.8Hz, 1H), 3.15 (s, 3H), 2.30 (s, 3H). 13 C NMR (100 MHz, CDCl3) δ: 161.9, 157.1, 154.4, 138.6, 137.9, 135.1, 131.2, 130.3, 129.9, 129.4 (2×C), 129.3, 128.8 (2×C), 128.6 (2×C), 128.4, 127.7, 127.1, 126.1, 125.5 (2×C), 121.2, 35.8, 21.4. Minor conformational isomers: 1 HNMR(400MHz, CDCl3)δ:7.47-7.31(m,10H),7.25-7.14(m,4H),7.06(d,J=9.2Hz,1H),6.22(d,J=9.2Hz,1H),3.31(s,3H),2.32(s,3H). 13 C NMR (100 MHz, CDCl3) δ: 162.6, 157.0, 154.6, 138.7, 137.9, 135.6, 131.3, 130.3, 129.9, 129.5 (2×C), 129.3, 129.0 (2×C), 128.6 (2×C), 128.4, 127.5, 127.1, 126.1, 125.5 (2×C), 120.1, 38.7, 21.4. HRMS (ESI) m / z calculated for C 25 H 23 ON4 + [M+H] + 395.1866, the measured value is 395.1874.

[0108] Example 21 Synthesis of Compound 2140:

[0109] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 4-chlorobenzaldehyde to obtain compound 2140 with a yield of 83%. The main conformational isomers were: 1H NMR (400MHz, CDCl3) δ: 7.47-7.29 (m, 13H), 7.25-7.21 (m, 1H), 6.90 (d, J = 8.8Hz, 1H), 6.05 (d, J = 8.8Hz, 1H), 3.15 (s, 3H). 13 C NMR (100MHz, CDCl3) δ: 161.7, 157.2, 153.2, 137.6, 136.7, 135.0, 130.4 (2×C), 130.2, 129.7, 129.6 (2×C), 129.5, 129.0 (2×C), 128.7 (2×C), 128.6 (2×C), 127.7, 125.5 (2×C), 121.5, 35.8. Minor conformational isomers: 1 H NMR (400MHz, CDCl3) δ: 7.47-7.29 (m, 13H), 7.25-7.21 (m, 1H), 7.05 (d, J = 9.2Hz, 1H), 6.22 (d, J = 9.2Hz, 1H), 3.30 (s, 3H). 13 C NMR (100 MHz, CDCl3) δ: 162.3, 157.1, 153.5, 137.6, 136.9, 135.5, 130.4 (2×C), 130.2, 129.7, 129.6 (2×C), 129.5, 129.3 (2×C), 128.7 (2×C), 128.4 (2×C), 127.5, 125.7 (2×C), 120.3, 38.7. HRMS (ESI) m / z calculated for C 24 H 20 ON4Cl + [M+H] + 415.1320, the measured value is 415.1313.

[0110] Example 22 Synthesis of Compound 2141:

[0111] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 3-chlorobenzaldehyde to obtain compound 2141 with a yield of 85%. The main conformational isomers were: 1 H NMR(400MHz, CDCl3)δ:7.57(s,1H),7.48-7.41(m,3H),7.39-7.28(m,7H),7 .26-7.21(m,3H),6.89(d,J=8.8Hz,1H),6.06(d,J=8.8Hz,1H),3.16(s,3H). 13C NMR (100 MHz, CDCl3) δ: 161.6, 157.2, 152.8, 137.5, 135.0, 134.7, 130.5, 130.1, 129.9, 129.7, 129.6 (2×C), 129.3 (2×C), 128.7 (2×C), 128.6 (2×C), 127.7, 127.1, 125.5 (2×C), 121.6, 35.8. Minor conformational isomers: 1 H NMR(400MHz, CDCl3)δ:7.62(s,1H),7.48-7.41(m,3H),7.39-7.28(m,7H),7 .26-7.21(m,3H),7.05(d,J=9.2Hz,1H),6.22(d,J=9.2Hz,1H),3.30(s,3H). 13 C NMR (100 MHz, CDCl3) δ: 162.2, 157.1, 153.1, 137.5, 135.5, 134.8, 130.7, 130.1, 129.9, 129.8, 129.6 (2×C), 129.0 (2×C), 128.9 (2×C), 128.4 (2×C), 127.5, 127.1, 125.5 (2×C), 120.3, 38.7. HRMS (ESI) m / z calculated for C 24 H 20 ON4Cl + [M+H] + 415.1320, measured value is 415.1327.

[0112] Example 23 Synthesis of Compound 2142:

[0113] The synthesis steps were the same as those in Example 2. The aldehyde reagent in the first step was 4-trifluoromethylbenzaldehyde to obtain compound 2142 with a yield of 85%. The main conformational isomers were: 1 H NMR(400MHz, CDCl3)δ:7.68-7.57(m,4H),7.52-7.43(m,3H),7.39-7.29(m,6H) ,7.26-7.22(m,1H),6.89(d,J=8.8Hz,1H),6.07(d,J=8.8Hz,1H),3.17(s,3H). 13C NMR (100MHz, CDCl3) δ: 161.5, 157.4, 152.8, 137.5, 135.0, 132.2 (q, J = 32.6Hz), 130.7, 130.1, 129.8, 129.7 (2×C), 1 29.5(2×C),128.7(2×C),128.6(2×C),127.8,125.7(q,J=3.7Hz,2×C),125.6(2×C),123.7(q,J=271Hz),121.8,35.8. 19 F NMR (376MHz,CDCl3)δ:-63.0(3×F).Minor conformational isomers: 1 H NMR(400MHz, CDCl3)δ:7.68-7.57(m,4H),7.52-7.43(m,3H),7.39-7.29(m,6H) ,7.26-7.22(m,1H),7.05(d,J=9.2Hz,1H),6.24(d,J=9.2Hz,1H),3.31(s,3H). 13 C NMR (100MHz, CDCl3) δ: 162.1, 157.3, 153.1, 137.5, 135.5, 132.2 (q, J = 32.6Hz), 130.6, 130.0, 129.9 (2×C), 129.8, 1 29.3(2×C),129.0(2×C),128.4(2×C),127.6,125.7(q,J=3.7Hz,2×C),125.6(2×C),123.7(q,J=271Hz),120.5,38.7. 19 F NMR (376 MHz, CDCl3) δ: -63.0 (3×F). HRMS (ESI) m / z calculated value is C 25 H 20 ON4F + [M+H] + 449.1584, the measured value is 449.1589.

[0114] Effect verification example 1 Screening of cytotoxicity and anti-inflammatory activity of 1,2,4-triazole-cis-enamide compounds

[0115] 1. Cell Culture

[0116] Mouse mononuclear macrophage Raw 264.7 cells were cultured in DMEM high glucose medium (Gibco, Catalog No.: 11995065) supplemented with 10% (V / V) FBS (Gibco, Catalog No.: 10270-106), 100 U / mL penicillin, and 100 μg / mL streptomycin (Gibco, Catalog No.: 15140122). The culture conditions were 37°C, 5% CO2, and the cells were passaged when they reached 80% confluence.

[0117] 2. Cytotoxicity Assay

[0118] Mouse mononuclear macrophage Raw 264.7 cells were used and 2×10 4 / mL, inoculated in a 96-well plate at a density of 100μL / well. When the cell coverage rate was about 80%, all synthetic compounds (10μM) were added, with 3 replicates in each group, and a blank control group was set up. After continued cultivation for 24h, 10μL CCK8 solution was added to each well, and the cells were continued to be incubated in a 37℃, 5% CO2 incubator for 2h. The absorbance OD value was measured at a wavelength of 450nm with an enzyme reader and the cell inhibition rate was calculated.

[0119] 3. Detection of the expression level of inflammatory factors by compounds

[0120] Raw 264.7 cells were cultured at 2×10 5 / mL, 1mL / well density was inoculated in 12-well plates. When the cell coverage rate was about 80%, positive control drug (dexamethasone, final concentration 1μM) and candidate compound (DMSO solvent control, candidate compound 10μM) were added. After 1 hour, LPS with a final concentration of 100ng / mL was added. LPS stimulation continued for 4 hours.

[0121] (1) RNA extraction and qPCR determination

[0122] After completely discarding the culture medium from the cultured cells, 0.5 mL of RL in the RNA extraction kit was added to each well for lysis; the cell lysate was collected and Total RNA was extracted according to the kit instructions (Vazyme, catalog number: RC112), and its concentration was measured using an ultra-micro UV-visible spectrophotometer;

[0123] (2) Reverse transcription reaction

[0124] A 20 μL reverse transcription system was established, including 4 μL 5× Buffer, X μL Total RNA (500 ng), and DEPC-H2O (16-X) μL in 8 tubes, which were marked with serial numbers and placed in a PCR instrument with the settings of 50°C, 15 min; 85°C, 5 s; and 16°C∝ for reverse transcription;

[0125] (3) Fluorescence quantification

[0126] Add 80 μL DEPC-H2O to the transcribed cDNA to make 100 μL, centrifuge and mix, and store at -20°C for later use; multiply (SYBR 10 μL + DEPC-H2O 10 μL + primer 0.5 μL) by the number of samples, take 18 μL / well to a 96-well plate, add 2 μL cDNA to form a 20 μL system, and centrifuge and mix at 1200 rpm for 1 min; place the sample plate in the CFX Connect Real-Time System (real-time fluorescence quantitative PCR instrument) for detection, and perform detection according to the program of (95°C, 2 min; 95°C, 20 s; 57°C, 20 s; 72°C, 20 s) for a total of 39 cycles, 95°C, 1 min; 55°C, 30 s; 95°C, 30 s;

[0127] (4) Analysis

[0128] Adoption 2 -ΔΔCt The experimental results were analyzed by the following calculation formula: △Ct target gene = Ct target gene - Ct internal reference gene, △△Ct target gene = △Ct experimental group target gene - △Ct control group target gene. -ΔΔCt It indicates the expression fold of the target gene in the experimental group relative to that in the control group.

[0129] The primers are shown in Table 1 below:

[0130] Table 1 Primer sequences.

[0131]

[0132] The results are as follows Figure 1 As shown, the synthesized 1,2,4-triazole-cis-enamide compounds showed no obvious toxicity except 2133. The results of inflammation screening showed that most compounds could significantly inhibit the secretion levels of inflammatory factors IL-1β and IL-6, among which compound 2125 had the strongest in vitro anti-inflammatory activity and inhibited inflammatory expression in a concentration-dependent manner.

[0133] Effect verification example 2 Anti-inflammatory effect of compound 2125 on OVA-induced rhinitis mice

[0134] 1. Purpose of the experiment

[0135] The therapeutic effect of compound 2125 on the OVA-induced mouse rhinitis model was investigated.

[0136] 2. Experimental Materials

[0137] Experimental animals: 30 Balb / c male mice, age: 6-8 weeks; weight: about 18-22 g, purchased from Zhuhai Baishitong Biological Co., Ltd.

[0138] Experimental instruments: heating stirrer, paraffin slicer, microscope, syringe, cell counter, balance, oven, refrigerated centrifuge, etc.

[0139] Experimental reagents: HE staining related reagents, PBS solution, etc.

[0140] 3. Animal Experiments

[0141] 1. Establishment of OVA-induced rhinitis mouse model: ① Modeling drug OVA (Sigma, catalog number: S7951); ② Modeling method Balb / c mice were used to establish a rhinitis model. 50μg OVA and 100μL 4% Al(OH)3 were dissolved in 100μL normal saline. 100μL of sensitization solution was injected intraperitoneally every 2 days for 14 days. 5% OVA was administered intranasally for 7 consecutive days from day 15 to day 22, and the mice in the control group were treated with an equal amount of normal saline.

[0142] 2. Animal grouping: blank (Control) group, model (AR) group, positive drug group (dexamethasone, Dex), treatment group (compounds 2125-L, 2125-M, 2125-H, three doses: 50 mg / kg, 100 mg / kg, 150 mg / kg), 5 animals in each group.

[0143] 3. Animal administration: Low (50 mg / kg), medium (100 mg / kg), and high (150 mg / kg) doses of compound 2125 were administered by gavage. Dexamethasone was used as the positive control group. The Control group and AR group were administered with an equal volume of normal saline by gavage once a day for 7 consecutive days. Blood was collected from the eyeballs after 24 hours, and the animals were euthanized by intraperitoneal injection of an overdose of 3% sodium pentobarbital.

[0144] 4. Animal sampling: After the mice were killed, nasal lavage fluid and nasal mucosal tissue were collected.

[0145] 5. Index observation: total number of inflammatory cells, pathology of mouse nasal mucosa tissue, expression of inflammation, etc.

[0146] (1) Behavioral scoring: The mice were observed 30 minutes after the last injection of OVA sensitizer, and the allergic symptoms of the mice were recorded, such as the number of sneezes, the number of nose scratching, and the runny nose. Mice with a score of >5 were considered to have successfully established the model. The specific scoring criteria are shown in Table 2. Figure 2 As shown in the figure, compared with the Control group, mice in the AR group showed rhinitis symptoms such as scratching the nose, sneezing, and runny nose. After administration, the rhinitis symptoms of mice were alleviated.

[0147] Table 2 Behavioral scoring criteria

[0148]

[0149] (2) The total number of cells in the mouse nasal lavage fluid was measured using a cell counter. The results are as follows Figure 3 As shown, compared with the blank group, the total number of cells in the nasal lavage fluid of mice in the AR group was significantly increased; compared with the AR group, Dex and high-dose compound 2125 reduced the total number of cells in the nasal lavage fluid.

[0150] (3) Giemsa staining

[0151] ①Add 70% ethanol and fix for 10 minutes.

[0152] ② Add an appropriate amount of prepared modified Giemsa staining working solution (1X) to the smear sample and stain for 45 minutes.

[0153] ③ Wash thoroughly with distilled water from one side, and after drying, observe and photograph under a microscope.

[0154] The results are as follows Figure 4 As shown in the figure, compared with the Control group, the number of cells in the nasal lavage fluid of mice in the AR group increased, and the number of eosinophils and mast cells increased. Each drug-treated group can effectively reduce the inflammatory cells in the nasal cavity.

[0155] (4) Detection of serum histamine levels by enzyme-linked immunosorbent assay

[0156] Serum collection: Collect whole blood of mice into 1.5 mL EP tubes, centrifuge at 4000 rpm for 10 min, and then centrifuge at 10000 rpm for 10 min. Take the supernatant for ELISA detection of cytokines.

[0157] The secretion of mouse HIS in serum was detected using an enzyme-linked immunosorbent assay (ELISA) kit (Jianglai, catalog number: Histamine HIS: JL45802). The specific steps are as follows:

[0158] a. Reagent Preparation

[0159] ① Dilute the washing solution (20×) to 1× with double distilled water to prepare the required washing solution. ② Add the standard diluent to 1 bottle of standard according to the volume marked on the standard label and incubate at room temperature for 15 minutes. ③ Take 5 clean 1.5 ml centrifuge tubes, add 500μL of standard diluent to each tube in advance, and dilute the standard in multiples to obtain seven standard concentrations of 50, 25, 12.5, 6.25, 3.12, 1.56, and 0.78pg / mL. Finally, add the diluted standards to the pre-coated plate wells in turn, and add the standard diluent directly as 0pg / mL concentration, for a total of seven standard concentrations. ④ Add 200μL to each well, and proceed to the next wash after about 15-30s. Wash the plate five times in total and pat dry on paper.

[0160] b. Operation steps

[0161] ① Take out the required strips from the aluminum foil bag after equilibration at room temperature for 10 minutes, and seal the remaining strips in a ziplock bag and return them to 4°C.

[0162] ② Sample addition: Add 50 μL of samples or standards of different concentrations to the corresponding wells, add 50 μL of universal diluent to the blank wells, and then add 50 μL of Biotin-antibody working solution to each well. Cover the plate with a sealing film and incubate at 37°C for 1 hour.

[0163] ③ Wash the plate 3 times: discard the liquid, add 300μL washing solution to each well, let it stand for 1 min, shake off the washing solution, pat dry on absorbent paper, and repeat washing the plate 3 times.

[0164] ④ Add enzyme conjugate working solution: Add 100 μL of enzyme conjugate working solution to each well, cover with sealing film and incubate at 37°C for 30 minutes.

[0165] ⑤ Wash the plate 5 times and pat dry on thick absorbent paper for the last wash.

[0166] ⑥ Add substrate: Add 90 μL of substrate (TMB) to each well, cover with sealing film, and incubate at 37°C in the dark for 15 min.

[0167] ⑦ Add stop solution: Add 50 μL of stop solution to each well and immediately measure the OD value of each well at a wavelength of 450 nm.

[0168] The results are as follows Figure 5 As shown, compared with the AR group, compound 2125 significantly downregulated the HIS level at doses of 50 mg / kg, 100 mg / kg, and 150 mg / kg.

[0169] (5) Detection of serum ovalbumin-specific immunoglobulin E and histamine levels by enzyme-linked immunosorbent assay

[0170] Serum collection: Collect whole blood of mice into 1.5 mL EP tubes, centrifuge at 4000 rpm for 10 min, and then centrifuge at 10000 rpm for 10 min. Take the supernatant for ELISA detection of cytokines.

[0171] The secretion of mouse IgE in serum was detected using an enzyme-linked immunosorbent assay (ELISA) kit (Jianglai, catalog number: mouse ovalbumin-specific IgE (OVA-sIgE): JL23405). The specific steps are as follows:

[0172] a. Reagent Preparation

[0173] ① Preparation of positive and negative control working solutions: Add 1mL of universal diluent to the freeze-dried control respectively, let it stand for 15 minutes until it is completely dissolved, and then gently mix. ② Preparation of HRP-secondary antibody working solution: Centrifuge the concentrated HRP-antibody at 1000×g for 1 minute 15 minutes before use, and dilute the 100× concentrated HRP-antigen to 1× working concentration with universal diluent. ③ Preparation of 1× washing solution: Take 10mL of 20× washing solution to 190mL of distilled water. ④ Washing: Add 200μL to each well, and proceed to the next time after about 15-30s. Wash the plate five times in total, and pat it dry on paper.

[0174] b. Operation steps

[0175] ① Take out the required strips from the aluminum foil bag after equilibration at room temperature for 10 minutes, and seal the remaining strips in a ziplock bag and return them to 4°C.

[0176] ② Sample addition: Add 100 μL of sample or positive and negative control into the corresponding wells respectively, add 10 μL of universal diluent into the blank wells, cover with sealing film and incubate at 37°C for 1 hour.

[0177] ③ Wash the plate 3 times: discard the liquid, add 300μL washing solution to each well, let it stand for 1 min, shake off the washing solution, pat dry on absorbent paper, and repeat washing the plate 3 times.

[0178] ④ Add secondary antibody: Add 100 μL of secondary antibody working solution to each well, cover with sealing film and incubate at 37°C for 30 min.

[0179] ⑤ Wash the plate 5 times and pat dry on thick absorbent paper for the last wash.

[0180] ⑥ Add substrate: Add 90 μL of substrate (TMB) to each well, cover with sealing film, and incubate at 37°C in the dark for 15 min.

[0181] ⑦ Add stop solution: Add 50 μL of stop solution to each well and immediately measure the OD value of each well at a wavelength of 450 nm.

[0182] The results are shown in Table 3. Compared with the AR group, compound 2125 at a dose of 150 mg / kg significantly reduced the positive expression of IgE.

[0183] Table 3

[0184]

[0185] (6) Tissue fixation, dehydration and embedding

[0186] ① Place the mouse tissues of each group in 10% neutral formaldehyde solution and fix for 24 hours.

[0187] ② Place the trimmed tissue block in an embedding box and wash it with running water for 24 hours to completely remove residual formaldehyde.

[0188] ③ Dehydrate the tissue in an alcohol gradient, with the following specific steps: 70% alcohol for 12 hours, 80% alcohol for 1.5 hours, 95% alcohol I for 45 minutes, 95% alcohol II for 30 minutes, 100% alcohol I for 25 minutes, and 100% alcohol II for 20 minutes.

[0189] ④ After dehydration, place the tissue in alcohol / xylene (1:1, v / v) solution for 20 minutes.

[0190] ⑤ Soak the tissue in xylene I for 20 minutes and then in xylene II for 10 minutes.

[0191] ⑥ Place the tissue in preheated melted paraffin I and paraffin II in an oven at 60-65℃ for 1 hour each.

[0192] ⑦ Pour a small amount of embedding paraffin into the preheated metal embedding frame, place the skin tissue block in it, perpendicular to the embedding frame, with the bottom flat, pour paraffin again, embed the tissue, and cool it.

[0193] ⑧ Paraffin section: Fix the tissue paraffin block on the Leica slicer, slice the slices to a thickness of 5 μM, and slice them continuously. Use toothless forceps to place the slices in 40℃ water for spreading, pick up the slices with a glass slide, bake the slices at 60℃ for 2h, and then store them in a slice box at room temperature for later use.

[0194] (7) HE staining

[0195] ① Dewaxing and hydration: Put the skin tissue paraffin sections into xylene I and xylene II for 15 minutes each, then put them into 100% ethanol I and ethanol II for 3 minutes each, 95% ethanol I and ethanol II for 3 minutes each, 80% ethanol for 3 minutes, and double distilled water for 1 minute;

[0196] ②Hematoxylin staining for 15 minutes, then wash off the excess stain on the slide;

[0197] ③1% hydrochloric acid ethanol (99mL 70% ethanol + 1mL concentrated hydrochloric acid) color separation for 3s, the cell nucleus and chromatin should be clear under the microscope;

[0198] ④ Rinse with running water for 15 minutes, and then with distilled water for 1 minute;

[0199] ⑤ Eosin for 2 minutes, rinse with running water for 1 minute;

[0200] ⑥ Dehydrate with 80% and 100% ethanol for 2s and 7min respectively;

[0201] ⑦ Xylene I and II for 5 min each;

[0202] ⑧ Sealing: Take the slide out of xylene II, add neutral gum to the tissue, gently cover with a coverslip, and let it dry naturally;

[0203] ⑨Observation: Observe pathological changes under a microscope, take photos and analyze.

[0204] like Figure 6 As shown in the figure, the nasal mucosa of mice in the Control group showed no epithelial thickening, no vascular dilation and congestion, clear nasal mucosal structure, and no inflammatory cell infiltration. In the AR group, significant proliferation of nasal mucosal epithelial cells, obvious dilation of small blood vessels and edema of tissue spaces were observed, and inflammatory cell infiltration dominated by eosinophils appeared; the inflammation of nasal mucosal tissues in each drug-treated group was reduced to varying degrees.

[0205] (8) PAS staining

[0206] ① Paraffin sections were routinely dewaxed into distilled water.

[0207] ②According to the instructions of the Bio-Time PAS kit, oxidize with periodic acid solution: take out the periodic acid solution and equilibrate it to room temperature, add 100 μL of periodic acid solution to each sample, react in a wet box in the dark for 10 minutes, remove the periodic acid solution, immerse in distilled water and place on a shaker to wash for 5 minutes.

[0208] ③Schiff reagent staining: add 100μL Schiff reagent to each sample, place in a wet box, and place in a 37℃ oven to avoid light for staining for 30min-1h. Remove the staining solution, soak in distilled water and wash on a shaker for 5min. Note: The optimal staining time for different tissues may vary. Please adjust the staining time according to the actual staining effect.

[0209] ④Hematoxylin staining: Add 100 μL of hematoxylin staining solution to each sample and stain for 30 seconds. Remove the staining solution and rinse with distilled water for more than two times, 3 seconds each time, until the floating color is washed away. For smears, after they are properly dried, they can be sealed with sealing solution and observed under a microscope. Cells can be washed with distilled water to remove excess color and then photographed under a microscope.

[0210] ⑤ Rapid dehydration with 95% ethanol for 2-3 seconds, and dehydration with anhydrous ethanol twice, each time for 5 seconds.

[0211] ⑥ Clear with xylene twice, 2 min each time, and seal with neutral gum.

[0212] Microscopic observation showed (see Figure 7 ), the nasal mucosa of mice in the Control group showed no epithelial thickening, no vascular dilation and congestion, clear nasal mucosal structure, and no goblet cells. In the AR group, significant proliferation of nasal mucosal epithelial cells and increased expression of goblet cells were observed in mice; inflammation of nasal mucosal tissues in all drug-treated groups was reduced to varying degrees.

[0213] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A 1,2,4-triazole-cis-enamide compound and a pharmaceutically acceptable salt or solvate thereof, characterized in that: The structural formula is shown in formula (I): In formula (I), R is hydrogen, C6-C14 aryl, five- to fourteen-membered heteroaryl, C1-C10 cycloalkyl, five- to fourteen-membered heterocyclyl, C1-C6 straight-chain alkyl or C1-C6 branched-chain alkyl; The C6-14 aryl, five- to fourteen-membered heteroaryl, C6-10 cycloalkyl or five- to fourteen-membered heterocyclic group is substituted by one or more substituents selected from halogen, halogenated C1-C6 alkyl, carbonyl, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, carboxyl, cyano, hydroxyl, carbonyl, phenyl-SO2-, nitro, C1-C6 alkyloxycarbonyl; and the five- to fourteen-membered heteroaryl or five- to fourteen-membered heterocyclic group contains 1-3 heteroatoms selected from N, O and S.

2. The 1,2,4-triazole-cis-enamide compound and its pharmaceutically acceptable salt or solvate according to claim 1, characterized in that: The halogen is F, Cl, Br or I.

3. The 1,2,4-triazole-cis-enamide compound and its pharmaceutically acceptable salt or solvate according to claim 1, characterized in that: The 1,2,4-triazole-cis-enamide compound and its pharmaceutically acceptable salt or solvate are selected from any one of the following compounds:

4. The 1,2,4-triazole-cis-enamide compound and its pharmaceutically acceptable salt or solvate according to claim 1, characterized in that: The pharmaceutically acceptable salt is a pharmaceutically acceptable salt, including an inorganic acid salt, an organic acid salt, an alkyl sulfonate or an aryl sulfonate.

5. The 1,2,4-triazole-cis-enamide compound and its pharmaceutically acceptable salt or solvate according to claim 1, characterized in that: The solvate includes a solvate of the 1,2,4-triazole-cis-enamide compound and water, ethanol, isopropanol, ether, ethylene glycol, acetone or glycerol.

6. A method for preparing the 1,2,4-triazole-cis-enamide compound and its pharmaceutically acceptable salt or solvate as claimed in any one of claims 1 to 5, characterized in that: The following steps are involved: a) Dissolve the compounds of formula I and formula II in acetonitrile solution at a stoichiometric ratio of 1:1 and react for 1-3 hours to obtain a compound of formula III: b) dissolving the product compound of formula III obtained in step a) in dry dichloromethane, and dropping the mixture into a dry dichloromethane solution of NBS-dimethyl sulfide cooled to -78°C, reacting at -78°C for 0.5-2h, and then heating the temperature to 20-25°C for 2-5h to obtain a compound of formula IV; c) Compounds of formula V and formula VI are mixed in a stoichiometric amount of 1: 1.2 is dissolved in water, sodium carbonate is added, and the reaction is carried out for 2-5 hours to obtain a compound of formula VII; d) dissolving the compound of formula IV obtained in step b) and the compound of formula VII obtained in step c) in dry toluene at a stoichiometric ratio of 1:1.2, adding triethylamine, and reacting at 110° C.-130° C. for 1-3 h to obtain a compound of formula VIII; e) dissolving the product compound of formula VIII obtained in step d) in an ammonia methanol solution and reacting for 2-8 days to obtain a compound of formula IX; f) reacting the product compound of formula IX obtained in step e) with the compound of formula X at a stoichiometric ratio of 1.2:1 at 40° C.-80° C. for 0.5-3 h to obtain a compound of formula XI; g) reacting the product compound of formula XI obtained in step f) with sodium hydride at -20°C-5°C for 0.5-1h, then adding iodomethane at this temperature, and finally heating to 20°C-50°C for 2-6h to obtain a compound of formula XII, i.e., a compound of formula (I); wherein R is hydrogen, C6-C14 aryl, five- to fourteen-membered heteroaryl, C1-C10 cycloalkyl, five- to fourteen-membered heterocyclyl, C1-C6 straight-chain alkyl, or C1-C6 branched-chain alkyl; The C6-14 aryl, five- to fourteen-membered heteroaryl, C6-10 cycloalkyl or five- to fourteen-membered heterocyclic group is substituted by one or more substituents selected from halogen, halogenated C1-C6 alkyl, carbonyl, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, carboxyl, cyano, hydroxyl, carbonyl, phenyl-SO2-, nitro, C1-C6 alkyloxycarbonyl; and the five- to fourteen-membered heteroaryl or five- to fourteen-membered heterocyclic group contains 1-3 heteroatoms selected from N, O and S.

7. Use of the 1,2,4-triazole-cis-enamide compound and its pharmaceutically acceptable salt or solvate as claimed in any one of claims 1 to 5 in the preparation of anti-inflammatory drugs.

8. Use of the 1,2,4-triazole-cis-enamide compound and the pharmaceutically acceptable salt or solvate thereof as claimed in any one of claims 1 to 5 in the preparation of a drug for treating allergic rhinitis.

Images