Cinnamic acid-capsaicin-tacrine derivatives and their preparation methods and applications

Abstract

The present invention discloses a series of cinnamic acid-capsaicin-tacrine derivatives, their preparation methods, and applications, belonging to the field of pharmaceutical technology. The applicant's experimental results demonstrate that the derivatives described herein have good cholinesterase (AChE / BuChE) and BACE-1 inhibitory activity, and are expected to be used in the treatment of neurodegenerative diseases, particularly Alzheimer's disease, and possess excellent potential medicinal value.

Description

Technical Field

[0001] The invention relates to cinnamic acid-capsaicin-tacrine derivatives and a preparation method and application thereof, belonging to the technical field of medicine. Background Art

[0002] Alzheimer's disease (AD) is the leading cause of dementia and the only one of the top ten fatal diseases in the world that has no effective treatment. With care costs of approximately US$1 trillion annually, it is rapidly becoming one of the most expensive, deadly and burdensome diseases of this century. The number of people with dementia worldwide in 2019 was approximately 50 million, and is expected to triple to 152 million by 2050. Currently, the treatment of AD mainly relies on acetylcholinesterase (AChE) inhibitors (such as donepezil, galantamine, and rivastigmine) and N-methyl-D-aspartate (NMDA) receptor antagonists (such as memantine). However, these available drugs can only improve symptoms such as cognitive and memory impairment, and cannot delay or prevent the progression of AD. Despite the urgent need, the research and development of AD drugs has been difficult.

[0003] The pathogenesis of AD is complex and has not yet been fully deciphered. The currently recognized pathogenesis is low levels of acetylcholine (ACh), the gradual deposition of extracellular β-amyloid protein (Aβ) in the brain, and the aggregation of intracellular Tau protein, which leads to neuronal death and cognitive impairment. The main targets affecting the occurrence and deterioration of AD include acetylcholinesterase (AChE), butyrylcholinesterase (BuChE or BChE), β-amyloid protein, and Tau protein. The multi-target ligand (MTDL) strategy is an important direction of current research. Multi-target drugs can act on more than one pathological target at the same time, which may be more promising and more effective in regulating the progression of AD.

[0004] To date, designing drugs that target cholinesterase, responsible for ACh hydrolysis, to compensate for cholinergic deficits remains an effective strategy for AD. Cholinesterase (ChE) comprises two types: AChE and BuChE. Under normal circumstances, AChE is the primary enzyme responsible for the hydrolysis and metabolism of ACh (~90%). Studies have demonstrated that the PAS site of AChE promotes Aβ deposition, inducing the formation of stable AChE-Aβ complexes that further exacerbate neuronal toxicity. Furthermore, accumulating evidence suggests that AChE levels are only elevated in the early to mid-stages of AD, declining to 90% of normal values in the late stages, rendering AChE inhibitors ineffective. In contrast, BuChE levels increase with disease progression, reaching 120% of normal levels, replacing AChE in its critical role in catalyzing ACh hydrolysis. This fact has prompted a re-examination of BuChE as a valuable target or co-target for the treatment of AD. Based on these findings, dual inhibitors of AChE and BuChE at two sites would be highly desirable. Although inadequate in many respects, ChE inhibition remains one of the very few means to temporarily slow the progression of AD.

[0005] The deposition of Aβ peptide in the brain plays an early and critical role in the pathogenesis of AD. It is produced by amyloid precursor protein (APP) through the amyloid production pathway (mainly dependent on β-secretase and γ-secretase). 40 and Aβ 42 There are two major Aβ peptide isoforms in the brain, the latter showing a higher tendency to aggregate due to the hydrophobicity of its two terminal residues. 42 Aβ is the main component of amyloid plaques and has been shown to be neurotoxic and can cause damage to neurons. 42 The aggregates (oligomers) formed can initiate a pathogenic cascade, ultimately leading to neuronal loss and dementia. In the brain, BACE-1 is the major β-secretase, a transmembrane enzyme, and the key rate-limiting enzyme for the formation of Aβ. These studies suggest that inhibiting BACE-1 can reduce Aβ in the brain. 42 The emergence of this method is indeed a feasible strategy for developing ideal anti-AD drugs.

[0006] Tacrine was the first cholinesterase inhibitor approved by the FDA for the treatment of AD, but was withdrawn from the market due to liver toxicity. Despite this, due to its applicability to chemical modification, it has become a widely used scaffold for drug development. Cinnamic acid is a class of aromatic carboxylic acids (C6-C3) found in plants. It is considered to be the core pharmacophore for the development of AD and has a wide range of pharmacological properties such as antioxidant, anti-inflammatory, anti-Aβ aggregation and neuroprotection. Maria Digiacomo et al. found that cinnamic acid-tacrine derivatives not only have strong AChE / BuChE inhibitory activity, but also exhibit weak BACE-1 inhibitory activity (M. Digiacomo, Z. Chen, S. Wang, A. Lapucci, M. Macchia, X. Yang, J. Chu, Y. Han, R. Pi, S. Rapposelli, Synthesis and pharmacological evaluation of multifunctional tacrine derivatives against several disease pathways of AD, Bioorganic & medicinal chemistry letters, 25 (2015) 807-810.). However, there are currently no reports on the use of N-(4-hydroxy-3-methoxybenzyl)alkylamine from capsaicin to modify the carboxyl group of cinnamic acid and connect tacrine with alkylene chains of different lengths to synthesize a series of cinnamic acid-capsaicin-tacrine derivatives as multifunctional preparations of AChE / BuChE and BACE-1. Summary of the Invention

[0007] The technical problem to be solved by the present invention is to provide a series of cinnamic acid-capsaicin-tacrine derivatives with novel structures and significant cholinesterase and BACE-1 inhibitory activities, as well as preparation methods and applications thereof.

[0008] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0009] The cinnamic acid-capsaicin-tacrine derivative of the present invention is a compound having the structure shown in the following formula 7 or a pharmaceutically acceptable salt thereof:

[0010]

[0011] wherein R1 represents a hydrogen atom, a trifluoromethyl group, a methoxy group, a fluorine atom, a chlorine atom or a bromine atom; R2 represents a hydrogen atom or a methoxy group; or R1 R2 represents a bis(methyleneoxy) group; R3 represents a hydrogen atom or a methoxy group; and n = 3 to 8.

[0012] In the general structure of the above-mentioned cinnamic acid-capsaicin-tacrine derivative, each substituent is preferably as follows:

[0013] R1, R2 and R3 all represent hydrogen atoms, and n=4, 5, 6, 7 or 8;

[0014] R1 R2 represent a bis(methyleneoxy) group, R3 represents a hydrogen atom, and n=5 or 6;

[0015] R1 and R2 both represent methoxy groups, R3 represents a hydrogen atom, and n=3, 5 or 6;

[0016] R1, R2 and R3 all represent methoxy, n=3, 4, 5, 6, 7 or 8;

[0017] R1 represents a trifluoromethyl group, R2 and R3 both represent a hydrogen atom, and n=3;

[0018] R1 and R2 both represent fluorine atoms, R3 represents a hydrogen atom, and n=3;

[0019] R1 represents a chlorine atom, R2 and R3 both represent a hydrogen atom, and n=3;

[0020] R1 represents a bromine atom, R2 and R3 both represent a hydrogen atom, and n=3.

[0021] More preferably, R1, R2 and R3 all represent methoxy groups, and n=7; or R1 represents trifluoromethyl groups, and R2 and R3 all represent hydrogen atoms, and n=3.

[0022] The pharmaceutically acceptable salts of the cinnamic acid-capsaicin-tacrine derivatives involved in the present invention can specifically be the hydrochloride, hydrobromide, phosphate, sulfate, fumarate, salicylate, benzenesulfonate, pyruvate, acetate, mandelate, alkaline metal cation salt or ammonium cation salt of the structural compound represented by the above formula 7.

[0023] The preparation method of the cinnamic acid-capsaicin-tacrine derivative of the present invention comprises the following steps:

[0024] 1) placing a compound represented by the following formula 1 and a compound represented by the following formula 2 in an organic solvent and reacting them in the presence of a condensing agent and an alkaline reagent to obtain a compound represented by the following formula 5;

[0025] 2) taking the compound represented by the following formula 3 and the compound represented by the following formula 5, placing them in an organic solvent, adding an alkaline reagent to react, and obtaining a compound represented by the following formula 6;

[0026] 3) taking the compound represented by the following formula 4 and the compound represented by the following formula 6, placing them in an organic solvent, adding an alkaline reagent to react, and obtaining the compound represented by the following formula 7;

[0027]

[0028] In the above formulae, R1 represents a hydrogen atom, a trifluoromethyl group, a methoxy group, a fluorine atom, a chlorine atom or a bromine atom; R2 represents a hydrogen atom or a methoxy group; or R1 R2 represents a bis-methyleneoxy group; R3 represents a hydrogen atom or a methoxy group; and n = 3 to 8.

[0029] In the above preparation method, the organic solvent involved can be one or a combination of two or more selected from dichloromethane (DCM), 1,2-dichloroethane (DCE), chloroform, chlorobenzene, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), and acetonitrile; preferably, N,N-dimethylformamide or acetonitrile. The amount of the organic solvent used can be determined as needed, generally to a level sufficient to dissolve the raw materials to be reacted.

[0030] In the above preparation method, the alkaline reagent involved can be a conventional choice in the prior art. Preferably, the alkaline reagent involved is one or a combination of two or more selected from sodium carbonate, potassium carbonate, cesium carbonate, potassium tert-butoxide, sodium tert-butoxide, sodium hydride, potassium hydride, triethylamine, sodium hydroxide and potassium hydroxide, and more preferably sodium hydride, triethylamine or potassium carbonate. As for the amount of the alkaline reagent used, in step 1), it is preferably 2 to 3 times the molar amount of the compound of the structure shown in Formula 1, in step 2), it is preferably 3 to 5 times the molar amount of the compound of the structure shown in Formula 5, and in step 3), it is preferably 1 to 2 times the molar amount of the compound of the structure shown in Formula 6.

[0031] In step 1) of the above preparation method, the condensing agent is a conventional choice in the prior art, such as a carbodiimide condensing agent, an organophosphorus condensing agent, etc., preferably a carbodiimide condensing agent, such as dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), or 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), with EDCI being preferred. The amount of the condensing agent used is generally 1 to 2 times the molar amount of the compound represented by Formula 1. When the condensing agent is a carbodiimide condensing agent, a condensation activator is preferably also added. The selection and amount of the condensation activator are the same as those in the prior art. Specifically, the condensation activator can be HOBt, DMAP, HOAt, 4-PPY, etc., and the combination of EDCI and HOBt is a conventional combination. The amount of the condensation activator used is generally 0.8 to 1.2 times the molar amount of the condensing agent. The reaction is preferably carried out in an ice bath or at room temperature.

[0032] In steps 1) to 3) of the above preparation method, the reaction can be carried out with or without heating. When the alkaline reagent selected is sodium hydride or contains sodium hydride, the reaction is preferably carried out at room temperature or in an ice bath, and more preferably, the reaction can be carried out under the protection of an inert atmosphere (such as nitrogen). For other alkaline reagents, the reaction is preferably carried out under heating, and more preferably under reflux at 50-70°C.

[0033] The ratio of the raw materials in each step of the above preparation method is their stoichiometric ratio, and the reaction time of each step is tracked and monitored by thin layer chromatography until the reaction is complete.

[0034] The above method prepares a crude product of the target compound having the structure shown in Formula 7. Therefore, the method of the present invention further includes the step of purifying the crude target compound obtained. Specifically, it can be purified by existing conventional purification methods to improve the purity of the target compound, such as purifying the crude product by silica gel column chromatography. It is more preferred to extract the material obtained by the reaction and then perform silica gel column chromatography to reduce the burden on the silica gel column. Among them, the eluent used for elution during column chromatography is preferably a mixed solvent composed of dichloromethane and methanol. In the mixed solvent, the volume ratio of dichloromethane to methanol is preferably 50:1 to 15:1. If extraction is involved, the extractant is preferably dichloromethane, 1,2-dichloroethane or ethyl acetate (EA).

[0035] The intermediate product obtained in step 1) and step 2) can be used directly in the next step. However, from the perspective of increasing the yield of the target compound, it is preferred to purify the intermediate product obtained in step 1) and step 2) before proceeding to the next step. The purification operation is usually performed by silica gel column chromatography, and more preferably, the material obtained by the reaction is first extracted and then subjected to silica gel column chromatography. In the column chromatography, the eluent used for elution is preferably a mixed solvent composed of petroleum ether and ethyl acetate, or a mixed solvent composed of dichloromethane and ethyl acetate in a volume ratio of 1:1 to 1:2. If extraction is involved, the extractant is preferably dichloromethane, 1,2-dichloroethane, or ethyl acetate.

[0036] The applicant has discovered through experiments that the cinnamic acid-capsaicin-tacrine derivatives of the present invention have good AChE / BuChE and BACE-1 inhibitory activity. Based on this, the present invention also includes the use of the above-mentioned cinnamic acid-capsaicin-tacrine derivatives or pharmaceutically acceptable salts thereof in the preparation of drugs for treating or preventing Alzheimer's disease; further, the present invention also includes the use of the above-mentioned cinnamic acid-capsaicin-tacrine derivatives or pharmaceutically acceptable salts thereof in the preparation of cholinesterase inhibitors, or in the preparation of BACE-1 inhibitors.

[0037] Furthermore, the present invention also includes a pharmaceutical composition comprising a therapeutically effective dose of the above-mentioned cinnamic acid-capsaicin-tacrine derivative or a pharmaceutically acceptable salt thereof as an active ingredient, and at least one pharmaceutically acceptable carrier.

[0038] Compared to the prior art, the present invention provides a series of novel cinnamic acid-capsaicin-tacrine derivatives and their preparation methods. The applicant's experimental results demonstrate that the derivatives possess excellent cholinesterase (AChE / BuChE) and BACE-1 inhibitory activity, demonstrating their potential for treating neurodegenerative diseases, particularly Alzheimer's disease, and possessing excellent medicinal value. DETAILED DESCRIPTION

[0039] In order to better explain the technical solution of the present invention, the present invention is further described in detail below with reference to examples, but the embodiments of the present invention are not limited thereto.

[0040] Example 1: General preparation method of compounds 5a-5h

[0041]

[0042] The compound represented by Formula 1 (6.30 mmol, 1.0 equiv.), EDCI (8.19 mmol, 1.3 equiv.), HOBT (8.19 mmol, 1.3 equiv.), and Et3N (18.90 mmol, 3.0 equiv. of Et3N) were placed in DMF (20 mL) and stirred in an ice bath for 30 min. The compound represented by Formula 2 (7.56 mmol, 1.2 equiv.) was then added and stirred at room temperature for 12 h. After the reaction, the mixture was poured into an appropriate amount of 5 v / v% HCl solution and extracted with EA (30 mL x 3). The combined organic phases were washed sequentially with 5 w / w% NaHCO3 solution, water, and saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. A small amount of the resulting residue was purified by TLC (PE / EA: 1 / 2, v / v) for structural confirmation. The crude product was used in the next step without further purification. Specific characterization is as follows:

[0043] Compound 5a: yield 79%; white solid; 1H NMR (600MHz, DMSO-d6) δ8.87(s,1H),8.49(t,J=5.9Hz,1H),7.58–7.52(m,2H),7.46(d,J=15.8Hz,1H),7.41(t,J=7.1H z,2H),7.39–7.34(m,1H),6.87(d,J=1.9Hz,1H),6.74–6.66(m,3H),4.29(d,J=5.8Hz,2H),3.75(s,3H).HRMS(ESI):m / z calcd for C 17 H 18 NO3 + [M+H] + :284.1281; found:284.1300.

[0044] Compound 5b: yield 88%; white solid; 1 H NMR (600MHz, DMSO-d6) δ8.86(s,1H),8.36(t,J=5.8Hz,1H),7.37(d,J=15.7Hz,1H),7.13(d,J=1.6Hz,1H),7.06(dd,J=8.0,1.7Hz,1H),6.94(d, J=8.0Hz,1H),6.86(d,J=1.9Hz,1H),6.73–6.65(m,2H),6.51(d,J=15.7Hz,1H),6.06(s,2H),4.27(d,J=5.8Hz,2H),3.74(s,3H).HRMS(ESI):m / z calcd for C 18 H 18 NO5 + [M+H] + :328.1179; found:328.1176.

[0045] Compound 5c: yield 87%; white solid; 1H NMR (600MHz, DMSO-d6) δ8.86(s,1H),8.36(t,J=5.9Hz,1H),7.39(d,J=15.7Hz,1H),7.15(d,J=2.0Hz,1H),7.11(dd,J=8.3,2.0Hz,1H),6.98(d,J=8. 3Hz,1H),6.87(d,J=1.9Hz,1H),6.73–6.67(m,2H),6.57(d,J=15.7Hz,1H) ,4.27(d,J=5.8Hz,2H),3.78(d,J=5.5Hz,6H),3.75(s,3H).HRMS(ESI):m / z calcdfor C 19 H 22 NO5 + [M+H] + :344.1492; found:344.1504.

[0046] Compound 5d: yield 87%; white solid; 1 H NMR(600MHz,DMSO-d6)δ8.86(s,1H),8.40(t,J=5.9Hz,1H),7.40(d,J=15.7Hz,1H),6.88(d,J=14.4Hz,3H),6.73–6 .67(m,2H),6.67–6.61(m,1H),4.28(d,J=5.8Hz,2H),3.80(s,6H),3.75(s,3H),3.68(s,3H).HRMS(ESI):m / zcalcd for C 20 H 24 NO6 + [M+H] + :374.1598; found:374.1616.

[0047] Compound 5e: yield 83%; white solid; 1 H NMR(600MHz,DMSO-d6)δ8.90(s,1H),8.51(t,J=5.8Hz,1H),7.94–7.84(m,2H),7.72(d,J=7.8Hz,1H),7.65(t,J=7.8Hz ,1H),7.55(d,J=15.9Hz,1H),6.91–6.80(m,2H),6.75–6.66(m,2H),4.29(d,J=5.7Hz,2H),3.75(s,3H).HRMS(ESI):m / z calcd for C 18 H 17 F3NO3+ [M+H] + :352.1155; found:352.1148.

[0048] Compound 5f: yield 86%; white solid; 1 H NMR (600MHz, DMSO-d6) δ8.88(s,1H),8.50(t,J=5.8Hz,1H),7.66(ddd,J=12.0,7.8,2.0Hz,1H),7.50–7.39(m,3H),6.87(d,J=1.9Hz ,1H),6.72(d,J=8.0Hz,1H),6.69(dd,J=8.0,1.9Hz,1H),6.66(d,J=15.8Hz,1H),4.28(d,J=5.8Hz,2H),3.74(s,3H).HRMS(ESI):m / z calcd for C 17 H 16 F2NO3 + [M+H] + :320.1093; found:320.1088.

[0049] Compound 5g: yield 87%; white solid; 1 H NMR (600MHz, DMSO-d6) δ8.88(s,1H),8.49(t,J=5.8Hz,1H),7.63(d,J=2.1Hz,1H),7.53(dt,J=6.7,2.0Hz,1H),7.4 7–7.40(m,3H),6.87(d,J=1.8Hz,1H),6.78–6.65(m,3H),4.29(d,J=5.8Hz,2H),3.75(s,3H).HRMS(ESI):m / zcalcd for C 17 H 17 ClNO3 + [M+H] + :318.0891; found:318.0897.

[0050] Compound 5h: yield 87%; white solid; 1H NMR (600MHz, DMSO-d6) δ8.88(s,1H),8.48(t,J=5.8Hz,1H),7.77(d,J=1.9Hz,1H),7.60–7.54(m,2H),7.43(d,J=15.8Hz,1H),7.37(t,J =7.9Hz,1H),6.87(d,J=1.9Hz,1H),6.76–6.71(m,2H),6.69(dd,J=8.0,1.9Hz,1H),4.29(d,J=5.8Hz,2H),3.75(s,3H).HRMS(ESI):m / z calcd for C 17 H 17 BrNO3 + [M+H] + :362.0386; found:362.0384.

[0051] Example 2: Preparation of Compound 5a

[0052] Example 1 was repeated except that DCE was used instead of DMF, potassium carbonate was used instead of triethylamine, DCC was used instead of EDCI, and DMAP was used instead of HOBt.

[0053] The residue was purified by TLC (PE / EA: 1 / 2, v / v) to give a white solid in 20% yield. Characterization by H NMR, C NMR, and high-resolution mass spectrometry confirmed it to be compound 5a.

[0054] Example 3: General Preparation Method of Compounds 6a-6t

[0055]

[0056] Compounds 5a-5h (1.40 mmol, 1.0 equiv.) and K2CO3 (7.00 mmol, 5.0 equiv.) were placed in acetonitrile (10 mL) and stirred to dissolve. The compound of formula 3 (1.68 mmol, 1.2 equiv.) was added thereto, and the resulting mixture was stirred and refluxed at 70°C for 10-12 h (TLC reaction monitoring). After the reaction was complete, the mixture was concentrated to dryness, diluted with water, and extracted with EA (20 mL × 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE / EA: 1 / 1-1 / 2, v / v) to obtain light yellow solid compounds 6a-6t. Specific characterization is as follows:

[0057] Compound 6a: Yield 85%; pale yellow solid; 1H NMR(600MHz,DMSO-d6)δ8.54(t,J=5.9Hz,1H),7.60–7.53(m,2H),7.46(d,J=15.8Hz ,1H),7.43–7.39(m,2H),7.39–7.35(m,1H),6.95–6.88(m,2H),6.80(dd,J=8.2,2.0H z,1H),6.69(d,J=15.8Hz,1H),4.32(d,J=5.9Hz,2H),3.95(t,J=6.3Hz,2H),3.74(s, 3H),3.61(t,J=6.7Hz,2H),1.95(p,J=6.9Hz,2H),1.85–1.77(m,2H).HRMS(ESI):m / z calcd for C 21 H 25 BrNO3 + [M+H] + :418.1012; found:418.1000.

[0058] Compound 6b: yield 78%; pale yellow solid; 1 H NMR (600MHz, DMSO-d6) δ8.59(d,J=6.2Hz,1H),7.56(d,J=7.3Hz,2H),7.45(d,J=15.8H z,1H),7.43–7.35(m,3H),6.94–6.87(m,2H),6.79(dd,J=8.2,2.0Hz,1H),6.71(dd,J=1 5.8,2.8Hz,1H),4.32(d,J=5.9Hz,2H),3.92(t,J=6.5Hz,2H),3.74(s,3H),3.65(t,J=6 .6Hz,2H),1.77(q,J=7.1Hz,2H),1.74–1.68(m,2H),1.54–1.48(m,2H).HRMS(ESI):m / z calcd for C 22 H 27 BrNO3 + [M+H] + :432.1169; found:432.1262.

[0059] Compound 6c: yield 88%; pale yellow solid; 1H NMR(600MHz,DMSO-d6)δ8.54(t,J=5.9Hz,1H),7.59–7.53(m,2H),7.46(d,J=15.8Hz, 1H),7.43–7.34(m,3H),6.94–6.86(m,2H),6.80(dd,J=8.2,2.0Hz,1H),6.69(d,J=15. 8Hz,1H),4.32(d,J=5.8Hz,2H),3.91(t,J=6.5Hz,2H),3.74(s,3H),3.53(t,J=6.7Hz ,2H),1.88–1.74(m,2H),1.69(p,J=6.4Hz,2H),1.42(p,J=2.9Hz,4H).HRMS(ESI):m / z calcd for C 23 H 29 BrNO3 + [M+H] + :446.1325; found:446.1311.

[0060] Compound 6d: yield 76%; pale yellow solid; 1 H NMR (600MHz, DMSO-d6) δ8.60(t,J=5.9Hz,1H),7.56(d,J=7.5Hz,2H),7.46(d,J=15.8Hz,1H),7.41(t, J=7.2Hz,2H),7.37(d,J=6.9Hz,1H),6.92–6.87(m,2H),6.79(dd,J=8.2,1.9Hz,1H),6.71(dd,J=15.9 ,2.1Hz,1H),4.32(d,J=5.8Hz,2H),3.90(t,J=6.5Hz,2H),3.74(s,3H),3.52(t,J=6.8Hz,2H),1.79(p ,J=7.0Hz,2H),1.68(p,J=7.1Hz,2H),1.39(t,J=7.6Hz,4H),1.35–1.30(m,2H).HRMS(ESI):m / zcalcd for C 24 H 31 BrNO3 + [M+H] + :460.1482; found:460.1476.

[0061] Compound 6e: yield 83%; pale yellow solid; 1H NMR(600MHz,DMSO-d6)δ8.54(t,J=5.9Hz,1H),7.59–7.52(m,2H),7.46(d,J=15.8Hz,1H),7.41(dd ,J=8.2,6.4Hz,2H),7.39–7.35(m,1H),6.92–6.86(m,2H),6.79(dd,J=8.2,2.0Hz,1H),6.69(d,J= 15.8Hz,1H),4.32(d,J=5.8Hz,2H),3.90(t,J=6.6Hz,2H),3.73(s,3H),3.51(t,J=6.7Hz,2H),1.7 8(p,J=6.8Hz,2H),1.68(p,J=6.7Hz,2H),1.38(q,J=7.3Hz,4H),1.33–1.27(m,4H).HRMS(ESI):m / z calcd for C 25 H 33 BrNO3 + [M+H] + :474.1638; found:474.1621.

[0062] Compound 6f: Yield 81%; pale yellow solid; 1 H NMR (600MHz, DMSO-d6) δ8.48(s,1H),7.37(d,J=15.7Hz,1H),7.13(d,J=1.7Hz,1H),7.06(dd, J=8.1,1.7Hz,1H),6.94(d,J=8.0Hz,1H),6.92–6.87(m,2H),6.78(dd,J=8.2,2.0Hz,1H),6.58 –6.51(m,1H),6.06(s,2H),4.30(d,J=5.9Hz,2H),3.91(t,J=6.4Hz,2H),3.73(s,3H),3.65(t, J=6.6Hz,2H),1.77(p,J=6.8Hz,2H),1.71(p,J=6.7Hz,2H),1.54–1.48(m,2H).HRMS(ESI):m / z calcd forC 23 H 27 BrNO5 + [M+H] + :476.1067; found:476.1236.

[0063] Compound 6g: yield 75%; pale yellow solid; 1H NMR (600MHz, DMSO-d6) δ8.45(t,J=5.9Hz,1H),7.37(d,J=15.7Hz,1H),7.14(d,J=1.7Hz,1H),7.06 (dd,J=8.0,1.7Hz,1H),6.94(d,J=8.0Hz,1H),6.91–6.86(m,2H),6.78(dd,J=8.3,2.0Hz,1H),6.53 (d,J=15.7Hz,1H),6.06(s,2H),4.30(d,J=5.8Hz,2H),3.90(t,J=6.5Hz,2H),3.73(s,3H),3.53(t, J=6.7Hz,2H),1.81(t,J=6.9Hz,2H),1.69(p,J=7.6,6.5Hz,2H),1.45–1.38(m,4H).HRMS(ESI):m / z calcd for C 24 H 29 BrNO5 + [M+H] + :490.1224; found:490.1212.

[0064] Compound 6h: yield 72%; light yellow solid; 1 H NMR(600MHz,DMSO-d6)δ8.42(t,J=6.0Hz,1H),7.39(d,J=15.7Hz,1H),7.16(d,J=2.0Hz,1H) ,7.11(dd,J=8.3,2.0Hz,1H),6.98(d,J=8.3Hz,1H),6.93(d,J=7.9Hz,2H),6.80(dd,J=8.2, 2.0Hz,1H),6.57(d,J=15.7Hz,1H),4.31(d,J=5.8Hz,2H),4.03(t,J=5.8Hz,2H),3.78(d,J= 5.7Hz, 6H), 3.75 (s, 3H), 3.66 (t, J=6.5Hz, 2H), 2.17 (dt, J=47.5, 6.3Hz, 2H). HRMS (ESI): m / z calcd forC 22 H 27 BrNO5 + [M+H] + :464.1067; found:464.1049.

[0065] Compound 6i: Yield 81%; pale yellow solid; 1H NMR (600MHz, DMSO-d6) δ8.40(t,J=5.9Hz,1H),7.39(d,J=15.7Hz,1H),7.15(d,J=2.0Hz,1H),7.11(dd ,J=8.3,1.9Hz,1H),6.98(d,J=8.4Hz,1H),6.90(d,J=8.4Hz,2H),6.79(dd,J=8.2,1.9Hz,1H),6.57(d, J=15.7Hz,1H),4.31(d,J=5.8Hz,2H),3.91(t,J=6.4Hz,2H),3.78(d,J=5.5Hz,6H),3.74(s,3H),3.55( td,J=6.7,1.2Hz,2H),1.86(p,J=6.9Hz,2H),1.71(p,J=6.7Hz,2H),1.58–1.46(m,2H).HRMS(ESI):m / z calcd for C 24 H 31 BrNO5 + [M+H] + :492.1380; found:492.1367.

[0066] Compound 6j: yield 80%; light yellow solid; 1 H NMR (600MHz, DMSO-d6) δ8.41(t,J=5.9Hz,1H),7.40(d,J=15.7Hz,1H),7.15(d,J=2.0Hz,1H),7.11(dd ,J=8.3,2.0Hz,1H),6.98(d,J=8.3Hz,1H),6.92–6.87(m,2H),6.79(dd,J=8.3,2.0Hz,1H),6.57(dd,J= 15.7,1.4Hz,1H),4.31(d,J=5.8Hz,2H),3.90(t,J=6.5Hz,2H),3.78(d,J=5.8Hz,6H),3.74(s,3H),3.5 3(t,J=6.7Hz,2H),1.81(p,J=6.8Hz,2H),1.69(t,J=6.8Hz,2H),1.43(h,J=3.5Hz,4H).HRMS(ESI):m / z calcd for C 25 H 33 BrNO5 + [M+H] + :506.1537; found:506.1528.

[0067] Compound 6k: yield 84%; pale yellow solid;1 H NMR(600MHz,Chloroform-d)δ7.58(d,J=15.5Hz,1H),6.86(dd,J=4.6,2.0Hz,2H),6.72(s,2H),6.32(d,J=15.3Hz,1H),5.88(t,J=5.3H z,1H),4.50(d,J=5.6Hz,2H),4.14(t,J=5.9Hz,2H),3.86(s,9H),3.85(s,3H),3.62(t,J=6.4Hz,2H),2.40–2.30(m,2H).HRMS(ESI):m / z calcd for C 23 H 29 BrNO6 + [M+H] + :494.1173; found:494.1166.

[0068] Compound 6l: yield 76%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ7.57(dd,J=15.5,1.9Hz,1H),6.85–6.81(m,2H), 6.71(d,J=2.1Hz,2H),6.33(dd,J=15.5,2.1Hz,1H),6.01–5.92(m,1H),4.49(d d,J=5.9,2.4Hz,2H),4.05–3.97(m,2H),3.87–3.80(m,12H),3.48(td,J=6.6,2 .0Hz,2H),2.06(t,J=7.2Hz,2H),1.97(dq,J=12.1,6.6Hz,2H).HRMS(ESI):m / z calcd for C 24 H 31 BrNO6 + [M+H] + :508.1329; found:508.1323.

[0069] Compound 6m: yield 82%; pale yellow solid; 1H NMR(600MHz,DMSO-d6)δ8.45(t,J=5.9Hz,1H),7.40(d,J=15.7Hz,1H),6.95–6.8 5(m,4H),6.79(dd,J=8.2,2.0Hz,1H),6.64(d,J=15.7Hz,1H),4.31(d,J=5.8Hz,2 H),3.92(t,J=6.5Hz,2H),3.80(s,6H),3.74(s,3H),3.67(s,3H),3.55(t,J=6.7 Hz,2H),1.90–1.80(m,2H),1.76–1.67(m,2H),1.60–1.45(m,2H).HRMS(ESI):m / z calcd for C 25 H 33 BrNO6 + [M+H] + :522.1486; found:522.1480.

[0070] Compound 6n: Yield 69%; pale yellow solid; 1 H NMR(600MHz,DMSO-d6)δ8.44(t,J=5.9Hz,1H),7.40(d,J=15.7Hz,1H),6.92–6.87(m ,4H),6.79(dd,J=8.2,2.0Hz,1H),6.64(d,J=15.7Hz,1H),4.31(d,J=5.8Hz,2H),3. 91(t,J=6.5Hz,2H),3.80(s,6H),3.74(s,3H),3.68(s,3H),3.53(t,J=6.7Hz,2H),1 .85–1.78(m,2H),1.69(p,J=6.6Hz,2H),1.43(p,J=3.4Hz,4H).HRMS(ESI):m / zcalcd for C 26 H 35 BrNO6 + [M+H] + :536.1642; found:536.1622.

[0071] Compound 6o: yield 87%; pale yellow solid; 1H NMR(600MHz,Chloroform-d)δ7.57(d,J=15.5Hz,1H),6.84–6.82(m,2H),6.71(s,2H),6.32(d,J=15.5Hz,1H),5.90(t,J=5.6Hz,1H),4.49(d,J=5.6Hz,2H ),3.99(t,J=6.7Hz,2H),3.85(d,J=7.2Hz,12H),3.40(t,J=6.8Hz,2H),1.85 (d,J=7.3Hz,4H),1.50–1.43(m,4H),1.41–1.35(m,2H).HRMS(ESI):m / zcalcd for C 27 H 37 BrNO6 + [M+H] + :550.1799; found:550.1793.

[0072] Compound 6p: yield 85%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ7.57(d,J=15.5Hz,1H),6.87–6.80(m,3H),6.72(s ,2H),6.32(d,J=15.5Hz,1H),4.50(d,J=5.6Hz,2H),3.99(t,J=6.8Hz,2H),3.86 (d,J=7.3Hz,12H),3.40(t,J=6.8Hz,2H),1.83(ddd,J=15.0,12.9,7.1Hz,4H),1 .44(dq,J=13.9,6.9Hz,4H),1.36(ddd,J=13.1,9.4,4.9Hz,4H).HRMS(ESI):m / z calcd for C 28 H 39 BrNO6 + [M+H] + :564.1955; found:564.1951.

[0073] Compound 6q: Yield 84%; pale yellow solid; 1H NMR (600MHz, DMSO-d6) δ8.57(t,J=5.8Hz,1H),7.92(s,1H),7.88(d,J=7.7Hz,1 H),7.73(d,J=7.8Hz,1H),7.65(t,J=7.8Hz,1H),7.55(d,J=15.9Hz,1H),6.96– 6.92(m,2H),6.85–6.79(m,2H),4.33(dd,J=5.8,2.8Hz,2H),4.04(t,J=6.0Hz, 2H),3.75(s,3H),3.66(t,J=6.5Hz,2H),2.21(t,J=6.3Hz,2H).HRMS(ESI):m / z calcd for C 21 H 22 BrF3NO3 + [M+H] + :472.0730; found:472.0712.

[0074] Compound 6r: Yield 80%; pale yellow solid; 1 H NMR(600MHz, DMSO-d6)δ8.55(q,J=4.5,3.0Hz,1H),7.67(ddd,J=11.9,7.8,2.0Hz,1H),7.50–7.42(m,3H),6.95–6.90(m,2H),6.82–6.77(m,1H) ,6.67(d,J=15.8Hz,1H),4.32(d,J=5.8Hz,2H),4.03(t,J=6.0Hz,2H),3.75(s,3H),3.65(t,J=6.5Hz,2H),2.21(t,J=6.3Hz,2H).HRMS(ESI):m / z calcd forC 20 H 21 BrF2NO3 + [M+H] + :440.0667;found:440.0680.

[0075] Compound 6s: yield 78%; pale yellow solid; 1H NMR(600MHz, DMSO-d6)δ8.54(t,J=5.9Hz,1H),7.64(d,J=2.1Hz,1H),7.56–7.51(m,1H),7.46–7.42(m,3H),6.96–6.90(m,2H),6.84–6.78(m,1H),6 .74(d,J=15.8Hz,1H),4.33(dd,J=5.8,3.0Hz,2H),4.04(t,J=5.9Hz,2H),3.75(s,3H),3.66(t,J=6.5Hz,2H),2.21(p,J=6.3Hz,2H).HRMS(ESI):m / z calcd forC 20 H 22 BrClNO3 + [M+H] + :438.0466; found:438.0490.

[0076] Compound 6t: yield 77%; pale yellow solid; 1 H NMR (600MHz, DMSO-d6) δ8.53(t,J=5.9Hz,1H),7.78(s,1H),7.57(t,J=8.0Hz,2H),7.46–7.35(m,2H),6.94(d,J=8.6Hz,2H),6.81(d,J=8.4Hz,1 H),6.74(d,J=15.8Hz,1H),4.33(d,J=5.8Hz,2H),4.04(t,J=6.0Hz,2H),3.75(s,3H),3.66(t,J=6.5Hz,1H),2.25–2.16(m,2H).HRMS(ESI):m / z calcd forC 20 H 22 Br2NO3 + [M+H] + :481.9961;found:482.0097.

[0077] Example 4: Preparation of Compounds 6a and 6t

[0078] Compound 6a: Example 3 was repeated, except that chlorobenzene was used instead of acetonitrile and sodium hydroxide was used instead of K2CO3. The reaction was carried out at 100°C until completion. The residue was purified by silica gel column chromatography (PE / EA: 1 / 1 to 1 / 2, v / v) to obtain a pale yellow solid in 62% yield. Characterization by H NMR, C NMR, and high-resolution mass spectrometry confirmed the product to be Compound 6a.

[0079] Compound 6t: Example 3 was repeated, except that DMF was used instead of acetonitrile and sodium hydride was used instead of K2CO3. The reaction was carried out at room temperature under nitrogen until completion. The residue was purified by silica gel column chromatography (PE / EA: 1 / 1 to 1 / 2, v / v) to obtain a pale yellow solid in 65% yield. Characterization by H NMR, C NMR, and high-resolution mass spectrometry confirmed the product to be Compound 6t.

[0080] Example 5: General Preparation Method of Compounds 7a-7t

[0081]

[0082]

[0083] Compound 4 (1.15 mmol, 1.2 equiv.) was placed in DMF (10 mL). NaH (1.92 mmol, 2.0 equiv., 60% mineral oil) was added under nitrogen and stirred in an ice bath for 30 min. Compounds 6a-6t (0.96 mmol, 1.0 equiv.) were then added, respectively, and the mixture was stirred at room temperature for 10 h. After completion of the reaction, an appropriate amount of ice water was added to quench the reaction, and the mixture was extracted with EA (20 mL × 1, 10 mL × 2). The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The resulting residue was purified by column chromatography (DCM / MeOH: 50 / 1 to 15 / 1, v / v) to give compounds 7a-7t. Specific characterization is as follows:

[0084] Compound 7a: yield 36%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.43(d,J=8.5Hz,1H),8.17(d,J=8.7Hz,1H),7.71–7.61(m,2H),7.56–7.50(m ,2H),7.43–7.36(m,2H),7.36–7.29(m,3H),6.83–6.72(m,3H),6.57(d,J=8.1Hz,1H),4.49(d,J=6.0Hz,2H) ,4.09(t,J=5.8Hz,2H),4.03(t,J=5.4Hz,2H),3.64(s,1H),3.61(s,3H),3.15(t,J=6.3Hz,2H),2.53(t,J=6 .3Hz,2H),2.02(p,J=6.2,5.6Hz,2H),1.93(p,J=6.0Hz,2H),1.81(q,J=6.5,5.9Hz,2H),1.77–1.73(m,2H). 13C NMR (150MHz, CDCl3) δ166.36,155.90,151.49,149.09,146.67,140.97,13 8.80,135.15,132.38,132.28,129.68,128.91,128.05,125.28,124.16,12 1.26,121.04,120.04,116.15,112.42,111.46,110.99,68.75,55.77,47. 83,43.35,32.07,29.84,28.38,25.63,22.83,21.99.HRMS(ESI):m / zcalcd forC 34 H 38 N3O3 + [M+H] + :536.2908; found:536.2965.

[0085] Compound 7b: yield 41%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.40(d,J=8.5Hz,1H),8.18(d,J=8.7Hz,1H),7.67–7.54(m,3H),7.50(dd,J=6.9,2.8Hz,2 H),7.39(ddd,J=8.4,7.0,1.2Hz,1H),7.30(dd,J=5.3,1.8Hz,3H),6.86(d,J=2.0Hz,1H),6.80–6.73(m,2H),6.47(d,J= 8.2Hz,1H),4.47(d,J=6.0Hz,2H),3.96(t,J=6.8Hz,2H),3.84(t,J=6.0Hz,2H),3.71(s,3H),3.63(s,1H),3.09(t,J=6. 3Hz,2H),2.55(t,J=6.3Hz,2H),1.90(p,J=6.9Hz,2H),1.79(q,J=6.5Hz,4H),1.74–1.69(m,2H),1.62(p,J=7.5Hz,2H). 13CNMR(150MHz,CDCl3)δ166.35,155.79,151.39,149.42,147.34,140.73,13 8.87,135.15,132.38,132.13,129.59,128.85,127.97,125.23,124.37,12 1.42,120.90,120.19,115.96,113.06,111.88,111.12,68.73,56.00,48.1 8,43.40,30.37,29.82,28.37,23.81,22.92,21.92,20.64.HRMS(ESI):m / z calcd for C 35 H 40 N3O3 + [M+H] + :550.3064; found:550.3123.

[0086] Compound 7c: yield 55%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.42(d,J=8.5Hz,1H),8.18(d,J=8.6Hz,1H),7.67–7.60(m,2H),7.54–7.45(m,2H),7. 43–7.36(m,2H),7.30(dd,J=5.1,2.1Hz,3H),6.84(d,J=2.0Hz,1H),6.79(dd,J=8.2,2.0Hz,1H),6.73(d,J=15.6Hz,1 H),6.60(d,J=8.2Hz,1H),4.48(d,J=5.8Hz,2H),3.93(t,J=6.9Hz,2H),3.86(t,J=6.3Hz,2H),3.67(s,3H),3.63(s, 1H),3.08(t,J=6.4Hz,2H),2.53(t,J=6.4Hz,2H),1.87–1.77(m,6H),1.72(dp,J=8.9,2.7Hz,2H),1.53–1.48(m,4H). 13C NMR (150MHz, CDCl3) δ166.28,155.67,151.45,149.47,147.53,140.80,139. 01,135.11,132.41,131.86,129.62,128.86,127.97,125.20,124.46,121.3 2,120.99,120.17,115.90,113.09,111.83,110.87,68.60,55.95,48.25,43 .46,30.70,29.82,28.78,28.35,25.80,25.27,21.90,20.65.HRMS(ESI):m / z calcd for C 36 H 42 N3O3 + [M+H] + :564.3221; found:564.3218.

[0087] Compound 7d: yield 35%; light yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.41(d,J=8.4Hz,1H),8.16(d,J=8.8Hz,1H),7.68–7.59(m,2H),7.49(dd,J=6.9, 2.8Hz,2H),7.41(t,J=7.8Hz,1H),7.32(d,J=5.0Hz,3H),6.98(s,1H),6.85–6.78(m,2H),6.69–6.59(m,2H),4.4 8(d,J=5.8Hz,2H),3.93–3.85(m,4H),3.76(s,3H),3.63(s,1H),3.14(t,J=6.3Hz,2H),2.56(t,J=6.5Hz,2H),1 .87(q,J=6.6Hz,2H),1.83(t,J=7.1Hz,2H),1.80–1.75(m,4H),1.47(dd,J=10.6,4.4Hz,4H),1.44–1.39(m,2H). 13C NMR (150MHz, CDCl3) δ166.17,155.13,152.14,149.56,147.71,140.90,135.0 6,132.02,131.62,129.65,128.86,127.93,127.90,125.06,124.28,121.74,1 21.17,120.22,116.32,113.19,111.81,111.36,69.10,56.03,48.51,43.55,3 0.90,28.92,28.83,28.58,26.36,25.91,23.85,22.05,20.91.HRMS(ESI):m / z calcd for C 37 H 44 N3O3 + [M+H] + :578.3377; found:578.3432.

[0088] Compound 7e: yield 31%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.41(dd,J=7.7,4.7Hz,1H),8.17(d,J=8.7Hz,1H),7.66–7.58(m,2H),7.47(dq,J=6.6,1.9Hz, 2H),7.43–7.37(m,1H),7.30(td,J=3.8,1.7Hz,3H),6.95(d,J=36.2Hz,1H),6.87–6.78(m,2H),6.70(dd,J=8.2,2.0Hz,1H), 6.66–6.57(m,1H),4.47(d,J=5.8Hz,2H),3.93–3.84(m,4H),3.77(d,J=1.5Hz,3H),3.63(s,1H),3.17(dt,J=8.7,3.9Hz,2H) ,2.58(t,J=6.3Hz,2H),1.88–1.83(m,2H),1.78(dq,J=20.0,7.2,6.8Hz,6H),1.45–1.40(m,4H),1.35(dd,J=7.4,3.8Hz,4H). 13C NMR (150MHz, CDCl3) δ166.24,154.59,151.73,149.46,147.62,141.57,138.89, 135.04,131.51,130.79,129.51,128.77,127.03,125.11,124.58,121.38,120. 66,119.72,115.86,113.05,111.37,110.87,69.01,55.56,48.35,43.44,30.43 ,29.76,29.12,29.01,28.94,28.43,26.50,25.75,21.96,20.69.HRMS(ESI):m / z calcd for C 38 H 46 N3O3 + [M+H] + :592.3534; found:592.3588.

[0089] Compound 7f: yield 32%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.41(d,J=8.5Hz,1H),8.14(d,J=8.7Hz,1H),7.69–7.61(m,1H),7.56(d,J=15.6Hz,1H),7.41 (ddd,J=8.4,6.8,1.2Hz,1H),7.23(t,J=6.0Hz,1H),7.07–6.97(m,2H),6.86(d,J=2.0Hz,1H),6.78–6.73(m,2H),6.60–6.4 2(m,2H),5.96(s,2H),4.48(d,J=6.0Hz,2H),3.97(d,J=7.1Hz,2H),3.87(t,J=6.0Hz,2H),3.74(s,3H),3.64(s,1H),3.11( t,J=6.3Hz,2H),2.54(t,J=6.4Hz,2H),1.90(q,J=7.3Hz,2H),1.84–1.80(m,4H),1.77–1.73(m,2H),1.66(q,J=7.2Hz,2H). 13C NMR (150MHz, CDCl3) δ166.49,155.76,151.47,149.44,149.06,148.29,147.36,1 40.65,138.86,132.49,132.17,129.54,125.29,124.32,123.93,120.98,120.23 ,119.30,115.89,113.08,111.88,111.07,108.59,106.67,101.52,68.74,56.03 ,48.27,43.44,32.06,29.83,28.36,23.69,22.83,21.90,20.65.HRMS(ESI):m / z calcd forC 36 H 40 N3O5 + [M+H] + :594.2962; found:594.3002.

[0090] Compound 7g: yield 51%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.41(d,J=9.0Hz,1H),8.18(d,J=8.3Hz,1H),7.68–7.58(m,1H),7.54(dt,J=13.8,3.1Hz,1H),7.44 –7.36(m,1H),7.24(d,J=9.1Hz,1H),7.03–6.92(m,2H),6.83(d,J=2.1Hz,1H),6.79(dd,J=8.0,2.1Hz,1H),6.74(dd,J=8.0,2.5H z,1H),6.60(dd,J=8.2,1.6Hz,1H),6.53(dd,J=15.6,3.3Hz,1H),5.95(d,J=1.7Hz,2H),4.47(d,J=5.0Hz,2H),3.95–3.84(m,4H) ,3.68(s,3H),3.63(s,1H),3.09(t,J=6.4Hz,2H),2.53(t,J=6.4Hz,2H),1.87–1.76(m,6H),1.75–1.70(m,2H),1.53–1.48(m,4H). 13C NMR (150MHz, CDCl3) δ166.48,155.68,151.33,149.43,148.99,148.24,147.50,14 0.49,139.00,132.34,131.86,129.51,125.15,124.52,123.86,120.87,120.12,11 9.37,115.93,113.04,111.78,110.96,108.53,106.59,101.49,68.62,55.95,48. 21,43.43,30.71,29.81,28.82,25.92,25.34,23.81,21.93,20.68.HRMS(ESI):m / z calcd for C 37 H 42 N3O5 + [M+H] + :608.3119; found:608.3173.

[0091] Compound 7h: yield 48%; light yellow solid; 1 H NMR (600MHz, Chloroform-d) δ8.44(d,J=8.5Hz,1H),8.11(t,J=8.9Hz,1H),7.66(d,J=1.6Hz,1H),7.62(d,J=15.6Hz,1H),7.51( s,1H),7.42(t,J=7.8Hz,1H),7.12(dq,J=3.6,1.9Hz,2H),6.83(d,J=8.8Hz,1H),6.79–6.75(m,1H),6.75–6.72(m,1H),6.53(d,J =8.2Hz,1H),6.25(s,1H),4.49(d,J=6.1Hz,2H),4.19(d,J=5.1Hz,2H),4.09(t,J=5.4Hz,2H),3.90(d,J=9.8Hz,6H),3.64(s,1H ),3.58(s,3H),3.12(t,J=6.4Hz,2H),2.59(t,J=6.2Hz,2H),2.26(t,J=5.7Hz,2H),1.81(d,J=6.0Hz,2H),1.74(t,J=5.7Hz,2H). 13C NMR (150MHz, CDCl3) δ166.83,156.80,151.24,150.47,149.14,149.00,146.58 ,140.52,138.81,132.98,132.22,128.20,125.02,124.68,122.03,120.60,11 9.80,119.44,116.28,112.91,111.85,111.33,111.08,109.93,67.34,56.06, 56.02,55.65,47.17,43.19,30.28,29.79,28.35,22.00,20.74.HRMS(ESI):m / z calcd for C 35 H 40 N3O5 + [M+H] + :582.2962;found:582.3022.

[0092] Compound 7i: yield 53%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.45(d,J=8.5Hz,1H),8.13(d,J=8.7Hz,1H),7.66(dd,J=8.6,6.8Hz,1H),7.61(d,J=15.6Hz,1H),7.44–7.39(m,1H ),7.24–7.19(m,1H),7.14–7.07(m,2H),6.86(d,J=2.0Hz,1H),6.84(d,J=8.2Hz,1H),6.75(dd,J=8.1,2.0Hz,1H),6.64(d,J=15.6Hz,1H),6.46( d,J=8.1Hz,1H),4.49(d,J=6.0Hz,2H),3.98(t,J=6.6Hz,2H),3.90(d,J =5.3Hz,6H),3.87–3.85(m,2H),3.74(d,J=2.0Hz,3H),3.64(d,1H),3.13 (t,J=6.4Hz,2H),2.54(t,J=6.4Hz,2H),1.91(p,J=6.8Hz,2H),1.82(dt,J=13.2,6.2Hz,4H),1.76(dd,J=8.1,3.7Hz,2H),1.68(q,J=7.4Hz,2H). 13C NMR (150MHz, CDCl3) δ166.59,155.72,151.77,150.50,149.40,149.15,147.34, 140.63,139.47,132.74,132.10,128.14,125.11,124.40,122.04,120.12,119.7 4,119.28,116.21,113.01,111.77,111.41,111.09,109.86,68.71,56.03,55.98 ,48.24,43.41,32.02,30.45,29.80,28.72,28.41,21.99,20.78.HRMS(ESI):m / z calcd forC 37 H 44 N3O5 + [M+H] + :610.3275; found:610.3336.

[0093] Compound 7j: yield 42%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.51(d,J=8.5Hz,1H),8.17(d,J=8.7Hz,1H),7.67(dd,J=8.5,6.9Hz,1H),7.60(d,J=15.6 Hz,1H),7.46–7.40(m,1H),7.14–7.06(m,3H),6.86–6.82(m,2H),6.80(dd,J=8.1,2.0Hz,1H),6.65–6.58(m,2H),4.50( d,J=5.9Hz,2H),3.96(t,J=6.8Hz,2H),3.89(d,J=4.4Hz,6H),3.87(d,J=6.2Hz,2H),3.67(s,3H),3.64(s,1H),3.12(t, J=6.4Hz,2H),2.51(t,J=6.4Hz,2H),1.89–1.83(m,4H),1.80(t,J=6.2Hz,2H),1.76–1.71(m,2H),1.54(q,J=3.6Hz,4H). 13C NMR (150MHz, CDCl3) δ166.56,155.63,150.42,149.38,149.08,147.44,140.44,139.0 4,137.80,132.17,131.83,128.13,125.05,124.54,121.96,120.85,120.00,119.37, 115.99,112.96,111.65,111.25,111.04,109.81,68.60,55.99,55.97,55.91,48.11, 43.36,30.67,29.76,28.81,28.47,25.93,25.34,21.94,20.68.HRMS(ESI):m / zcalcd for C 38 H 46 N3O5 + [M+H] + :624.3432; found:624.3486.

[0094] Compound 7k: yield 37%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.32(t,J=8.3Hz,1H),8.11(dd,J=8.6,4.0Hz,1H),7.66–7.55(m,3H),7.43–7.37 (m,1H),6.85–6.76(m,4H),6.73(dt,J=8.1,2.0Hz,1H),6.52(d,J=8.1Hz,1H),4.48(dd,J=6.1,2.0Hz,2H),4.1 4(d,J=5.7Hz,2H),4.08(td,J=5.5,2.3Hz,2H),3.86(dd,J=8.2,2.7Hz,9H),3.64(s,1H),3.58(s,3H),3.07(q, J=6.2,5.7Hz,2H),2.59(t,J=6.3Hz,2H),2.24(p,J=5.5Hz,2H),1.82–1.78(m,2H),1.74(dd,J=5.9,2.7Hz,2H). 13C NMR (150MHz, CDCl3) δ166.46,156.67,153.45,151.87,149.01,146.62,140. 63,139.42,139.23,133.00,132.14,130.92,125.05,124.45,121.02,120.5 9,119.91,116.51,112.85,112.04,111.44,105.11,70.67,61.06,56.31,55 .67,47.30,43.27,29.82,28.53,22.82,22.04,20.83.HRMS(ESI):m / zcalcd for C 36 H 42 N3O6 + [M+H] + :612.3068; found:612.3126.

[0095] Compound 7l: yield 40%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.40(d,J=8.5Hz,1H),8.19(d,J=8.7Hz,1H),7.64(t,J=7.8Hz,1H),7.59–7.49(m,2H) ,7.39(t,J=7.8Hz,1H),6.80(d,J=15.5Hz,4H),6.73(dd,J=8.1,2.0Hz,1H),6.53(d,J=8.1Hz,1H),4.48(d,J=6.0Hz ,2H),4.10(d,J=5.8Hz,2H),4.00(t,J=5.5Hz,2H),3.85(d,J=8.5Hz,9H),3.63(s,1H),3.59(s,3H),3.12(t,J=6.3H z,2H),2.55(t,J=6.3Hz,2H),2.02(t,J=6.5Hz,2H),1.92(t,J=6.1Hz,2H),1.83–1.79(m,2H),1.75(d,J=6.4Hz,2H). 13C NMR (150MHz, CDCl3) δ166.45,155.85,153.42,151.24,149.06,146.67,140. 61,139.41,138.79,132.21,132.10,130.87,125.18,124.34,120.97,120.7 5,119.90,116.17,112.40,111.31,111.09,105.09,68.62,61.04,56.28,55 .75,47.72,43.32,29.80,28.44,27.93,25.73,22.00,20.68.HRMS(ESI):m / z calcd for C 37 H 44 N3O6 + [M+H] + :626.3225; found:626.3281.

[0096] Compound 7m: yield 34%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.31(d,J=8.7Hz,1H),8.19(d,J=8.7Hz,1H),7.60(m,2H),7.53(dd,J=15.6, 4.3Hz, 1H), 7.37 (t, J = 7.7Hz, 1H), 6.83 (s, 1H), 6.74 (d, J = 15.9Hz, 4H), 6.47 (d, J = 8.1Hz, 1H), 4.45 (d, J = 5. 7Hz,2H),3.93(t,J=6.8Hz,2H),3.82(dd,J=5.3,3.1Hz,11H),3.70(d,J=3.3Hz,3H),3.62(s,1H),3.05(t,J =6.0Hz,2H),2.57(t,J=6.3Hz,2H),1.88(p,J=7.2Hz,2H),1.79(m,4H),1.72(m,2H),1.60(q,J=7.4Hz,2H). 13C NMR (150MHz, CDCl3) δ166.38,155.74,153.39,151.27,149.37,147.32,140.5 5,139.37,138.90,132.28,131.96,130.83,125.15,124.46,120.96,120.75,1 20.08,115.98,112.99,111.74,111.17,105.04,68.68,61.01,56.24,55.96,4 8.12,43.39,30.35,29.78,28.41,23.87,22.96,21.92,20.66.HRMS(ESI):m / z calcd for C 38 H 46 N3O6 + [M+H] + :640.3381;found:640.3436.

[0097] Compound 7n: yield 45%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.46–8.36(m,1H),8.18(d,J=8.7Hz,1H),7.66–7.59(m,1H),7.55(dd,J=15.6,2.3Hz,1H),7.4 2–7.28(m,2H),6.83(d,J=2.0Hz,1H),6.78(dd,J=8.3,2.0Hz,1H),6.75(d,J=2.6Hz,2H),6.70(d,J=15.6Hz,1H),6.59(d,J=8 .1Hz,1H),4.52–4.45(m,2H),3.93(t,J=6.8Hz,2H),3.86(d,J=5.9Hz,2H),3.85–3.82(m,9H),3.67(s,3H),3.63(s,1H),3.0 8(t,J=6.4Hz,2H),2.54(t,J=6.3Hz,2H),1.88–1.80(m,4H),1.77(d,J=6.1Hz,2H),1.75–1.69(m,2H),1.50(p,J=3.3Hz,4H). 13C NMR (150MHz, CDCl3) δ166.29,155.54,153.38,151.46,149.43,147.50,140.55, 139.37,139.20,132.18,131.76,130.82,125.08,124.49,121.00,120.92,120. 06,116.04,113.04,111.72,111.06,105.02,68.63,61.01,56.23,55.92,48.19 ,43.43,30.69,28.81,25.91,25.32,23.83,21.94,21.14,20.71.HRMS(ESI):m / z calcd forC 39 H 48 N3O6 + [M+H] + :654.3538; found:654.3578.

[0098] Compound 7o: yield 53%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.44(dd,J=8.6,1.3Hz,1H),8.19(d,J=8.7Hz,1H),7.62(ddd,J=8.3,6.9,1.2Hz,1H),7.55(d,J=15 .5Hz,1H),7.41(ddd,J=8.4,6.9,1.3Hz,1H),7.11(t,J=5.9Hz,1H),6.82(d,J=2.0Hz,1H),6.78(dd,J=8.2,2.0Hz,1H),6.74(s,2H ),6.66–6.61(m,2H),4.47(d,J=5.9Hz,2H),3.92(t,J=7.0Hz,2H),3.88(s,2H),3.84(s,9H),3.74(s,3H),3.63(s,1H),3.13(t,J= 6.4Hz,2H),2.56(t,J=6.4Hz,2H),1.85(td,J=10.5,8.2,4.0Hz,4H),1.80–1.74(m,4H),1.47(q,J=7.3Hz,4H),1.43–1.40(m,2H). 13C NMR (150MHz, CDCl3) δ166.17,155.51,153.46,151.63,149.59,147.69,140.85,1 39.50,139.10,132.49,131.76,130.77,125.29,124.33,121.20,120.69,120.20, 115.87,113.31,111.81,110.83,105.07,69.21,61.07,56.29,56.03,48.43,43.5 5,30.74,29.82,28.39,26.13,25.92,23.58,22.82,21.92,20.69.HRMS(ESI):m / z calcd for C 40 H 50 N3O6 + [M+H] + :668.3694; found:668.3749.

[0099] Compound 7p: yield 54%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.46(d,J=8.5Hz,1H),8.18(d,J=8.7Hz,1H),7.63(t,J=7.8Hz,1H),7.55(d,J =15.5Hz,1H),7.44–7.39(m,1H),6.85–6.78(m,3H),6.73(s,2H),6.70(d,J=8.1Hz,1H),6.57(d,J=15.5Hz, 1H),4.48(d,J=5.8Hz,2H),3.91(q,J=6.9Hz,4H),3.84(s,9H),3.77(s,3H),3.63(s,1H),3.19(t,J=6.4Hz, 2H), 2.56 (t, J=6.4Hz, 2H), 1.88 (m, 2H), 1.83–1.76 (m, 6H), 1.43 (dd, J=13.8, 5.2Hz, 4H), 1.39–1.35 (m, 5H). 13C NMR (150MHz, CDCl3) δ166.07,155.46,153.46,151.69,149.61,147.82,140.88,139 .53,139.25,132.37,131.47,130.71,125.23,124.32,121.30,120.61,120.23,115 .94,113.22,111.81,110.88,105.05,69.15,61.06,56.27,56.07,48.68,43.60,31 .06,29.82,29.05,28.96,28.86,26.44,25.75,23.69,21.98,20.75.HRMS(ESI):m / z calcd for C 41 H 52 N3O6 + [M+H] + :682.3851; found:682.3904.

[0100] Compound 7q: yield 38%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.33(d,J=8.6Hz,1H),8.08(d,J=8.6Hz,1H),7.79–7.73(m,2H),7.70(d,J=15.7Hz,2H ),7.65(t,J=7.7Hz,1H),7.57(d,J=7.8Hz,1H),7.48(t,J=7.8Hz,1H),7.42(t,J=7.8Hz,1H),6.92(t,J=15.7Hz,1H), 6.79–6.71(m,2H),6.53(d,J=8.1Hz,1H),4.49(d,J=6.0Hz,2H),4.11(dt,J=29.1,5.6Hz,4H),3.64(s,1H),3.58(s,3 H),3.08(t,J=6.4Hz,2H),2.59(t,J=6.3Hz,2H),2.23(dd,J=10.1,4.5Hz,2H),1.83–1.80(m,2H),1.77–1.74(m,2H). 13C NMR (150MHz, CDCl3) δ165.88,157.15,148.99,147.42,146.60,139.11,138.9 2,136.09,132.92,132.35,131.43,130.94,129.47,126.01,125.15,124.79, 124.76,124.42,123.40,121.02,119.93,116.42,112.82,111.88,111.47,67 .06,55.68,47.35,43.30,32.07,29.84,22.84,21.97,20.74.HRMS(ESI):m / z calcd for C 34 H 35 F3N3O3 + [M+H] + :590.2625;found:590.2684.

[0101] Compound 7r: yield 41%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.31(t,J=4.5Hz,1H),8.15(d,J=8.9Hz,2H),7.62(t,J=7.8Hz,1H),7.53(dd,J=15.9,2.8Hz,1H), 7.40(t,J=7.8Hz,1H),7.36–7.29(m,1H),7.26–7.22(m,1H),7.08(d,J=9.1Hz,1H),6.88(d,J=15.7Hz,1H),6.77(d,J=2.0Hz,1H) ,6.70(dd,J=8.1,2.0Hz,1H),6.48(d,J=8.1Hz,1H),4.46(d,J=6.0Hz,2H),4.17(t,J=5.7Hz,2H),4.04(t,J=5.4Hz,2H),3.63(s ,1H),3.56(s,3H),3.04(t,J=6.3Hz,2H),2.59(t,J=6.2Hz,2H),2.26(q,J=5.6Hz,2H),1.81–1.75(m,2H),1.72(q,J=6.0Hz,2H). 13C NMR (150MHz, CDCl3) δ166.06,156.99,151.23,148.96,146.55,138.74,138 .21,132.83,132.30,125.43,125.10,124.67,124.59,122.86,120.55,119 .83,117.72,117.61,116.30,116.28,116.16,112.77,111.87,111.39,67. 10,55.66,47.19,43.19,30.36,29.81,28.27,21.97,20.72.HRMS(ESI):m / z calcd for C 33 H 34 F2N3O3 + [M+H] + :558.2563; found:558.2617.

[0102] Compound 7s: yield 18%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.33(d,J=7.7Hz,1H),8.10(d,J=8.6Hz,1H),7.80(s,1H),7.68–7.57 (m,2H),7.49(s,1H),7.41(m,2H),7.27(d,J=2.0Hz,2H),6.84(d,J=15.7Hz,1H),6.79–6.71(m,2H), 6.52(d,J=8.1Hz,1H),4.48(d,J=6.0Hz,2H),4.15(t,J=5.5Hz,2H),4.08(t,J=5.4Hz,2H),3.64(s,1 H),3.58(s,3H),3.11–3.04(m,2H),2.59(t,J=6.3Hz,2H),1.80(t,J=5.8Hz,2H),1.76–1.72(m,2H). 13CNMR(150MHz,CDCl3)δ165.99,156.58,149.72,149.04,146.67,143.92,13 9.29,137.12,134.81,132.88,132.13,130.16,129.47,127.83,126.17,125 .06,124.38,122.89,121.44,119.96,116.80,112.91,112.14,111.51,67. 20,55.69,47.36,43.31,32.06,29.84,29.50,22.83,20.87.HRMS(ESI):m / z calcd for C 33 H 35 ClN3O3 + [M+H] + :556.2361; found:556.2428.

[0103] Compound 7t: yield 46%; pale yellow solid; 1 H NMR(600MHz,Chloroform-d)δ8.39(d,J=8.7Hz,1H),8.10(dd,J=8.9,3.6Hz,1H),7.81(t,J=6.8Hz,1H),7.70–7.63(m,2H) ,7.60(dd,J=15.8,3.4Hz,1H),7.50(d,J=7.8Hz,1H),7.44(d,J=7.8Hz,2H),7.21(t,J=7.9Hz,1H),6.85(d,J=15.6Hz,1H) ,6.77(d,J=2.0Hz,1H),6.73(d,J=8.1Hz,1H),6.51(d,J=8.1Hz,1H),4.49(d,J=6.1Hz,2H),4.19(s,2H),4.08(t,J=5.4Hz ,2H),3.64(s,1H),3.56(s,3H),3.09(t,J=6.1Hz,2H),2.58(t,J=6.3Hz,2H),2.25(m,2H),1.83–1.80(m,2H),1.74(m,2H). 13C NMR (150MHz, CDCl3) δ166.06,156.63,151.49,149.01,146.63,139.16,138 .81,137.44,132.75,132.23,132.02,130.68,130.35,126.49,124.96,124 .66,123.20,122.87,120.88,119.85,116.50,112.86,112.10,111.40,67. 26,55.67,47.11,43.22,30.33,29.79,28.59,22.06,20.84.HRMS(ESI):m / z calcd for C 33 H 35 BrN3O3 + [M+H] + :600.1856; found:600.1880.

[0104] Example 6: Preparation of Compounds 7a and 7t

[0105] Compound 7a: Example 5 was repeated, except that acetonitrile was used instead of DMF and potassium hydride was used instead of sodium hydride. The reaction was carried out at room temperature until completion. The resulting residue was purified by silica gel column chromatography (DCM / MeOH: 50 / 1 to 15 / 1) to obtain a pale yellow solid in a 25% yield. Characterization by H NMR, C NMR, and high-resolution mass spectrometry confirmed the product to be Compound 7a.

[0106] Compound 7t: Example 5 was repeated, except that DMSO was used instead of DMF. The resulting residue was purified by silica gel column chromatography (DCM / MeOH: 50 / 1 to 15 / 1) to afford a pale yellow solid in 21% yield. Characterization by H NMR, C NMR, and high-resolution mass spectrometry confirmed the product to be Compound 7t.

[0107] Experimental Example 1: Study on the AChE / BuChE inhibitory activity of target compounds 7a-7t

[0108] The inhibitory activity of the target compounds against AChE or BuChE was evaluated using Ellman spectrophotometry.

[0109] AChE (EC3.1.1.7, from electric eel and human), BuChE (EC3.1.1.8, from horse serum and human), 5,5′-dithiobis-(2-nitrobenzoic acid) (Ellman's reagent, DTNB), S-butylthiocholine iodide (BTCI), acetylthiocholine iodide (ATCI), and donepezil were purchased from Sigma-Aldrich. The specific steps were as follows: First, the test compound was dissolved in DMSO and diluted to the desired concentration (DMSO concentration less than 1%) with Tis-HCl buffer (pH = 8.0). 160 μL of DTNB (1.5 mM), 50 μL of AChE (0.22 U / mL), and 10 μL of the test drug were added sequentially to a 96-well plate and incubated at 37°C for 6 minutes. Then, 30 μL of ATCI (15 mM) was added as a substrate, and the absorbance change at 405 nm was measured (0, 60, 120, and 180 seconds) in a microplate reader (SpectraMax Plus 384, Molecular devices, CA, USA). Inhibition rate = [1-(absorbance change of experimental group / absorbance change of blank group)] * 100%. The determination of butyrylcholinesterase activity simply requires replacing AChE with BuChE and changing the substrate to thiobutyrylcholine (BTCI). Other procedures are the same. The molar concentration of the compound that provides 50% inhibition is calculated using Graph Pad Prism version 7.00 software (GraphPad Software, San Diego, CA), which is the compound's IC. 50 Each experiment was repeated three times, and the results are expressed as mean ± SD. The results are shown in Table 1.

[0110] Table 1. Inhibitory activity of compounds 7a-7t against cholinesterase

[0111]

[0112]

[0113] a The results are expressed as the mean (n=3) ± SD of three independent experiments.

[0114] The results showed that these compounds are strong cholinesterase (AChE / BuChE) inhibitors. Among the 7a~7t series of derivatives, compound 7t showed the strongest eeAChE inhibitory activity (eeAChE, IC 50=0.27 μM). Comparison of the results for compounds with different chain lengths shows that compounds containing three carbon spacers have better eeAChE inhibitory activity, while chain length is not conducive to eeAChE inhibition. Furthermore, the eeAChE inhibitory activity of compounds was studied by introducing different substituents at the 3-, 4-, and 5-positions of the cinnamic acid benzene ring. As shown in Table 1, when the substituent at the 3-position of the cinnamic acid benzene ring contains a halogen atom (7q-7t), the IC values for eeAChE inhibition are significantly higher than those for compounds containing halogen atom (7q-7t). 50 The values are all in the nanomolar range. The structure-activity relationship of cinnamic acid-capsaicin-tacrine derivatives in inhibiting eeAChE activity is as follows: halogen atom substitution (-CF3, -F, -Cl, -Br) on the cinnamic acid benzene ring ≈ three -OCH3 groups > two -OCH3 groups > no substituents > compounds containing OCH2O closed chains. In addition, all synthesized compounds have good inhibitory activity against eqBuChE, with IC 50 The values ranged from 0.09 μM to 0.44 μM, which were all more potent than the reference compound donepezil (eqBuChE, IC 50 =4.11 μM), among which compounds 7r and 7s had the strongest inhibitory activity against eqBuChE (IC 50 =0.09 μM). Variations in chain length and substituents had little effect on the inhibitory activity of eqBuChE, but overall, compounds with halogen atoms substituted on the cinnamic acid benzene ring (-CF3, -F, -Cl, -Br) were more potent.

[0115] In order to more accurately evaluate the cholinesterase activity of the target compound, human cholinesterase (hAChE / hBuChE) was selected to further determine the activity of the compound. As shown in Table 1, the effects of chain length and substituent changes on the inhibitory activity of hAChE were slightly different from those on eeAChE. Compound 7o (hAChE, IC 50 =0.04μM) has the strongest inhibitory activity against hAChE. The inhibitory activity of compounds 7a~7e and 7l~7p on hAChE by changing the length of the alkyl chain generally increases gradually with the extension of the carbon chain length. The structure-activity relationship of cinnamic acid-capsaicin-tacrine derivatives in inhibiting hAChE activity is as follows: halogen atom substitution (-CF3, -F, -Cl, -Br) on the cinnamic acid benzene ring ≈ three -OCH3 groups > no substituents > two -OCH3 groups > compounds containing OCH2O closed chain. In addition, the inhibitory activity of all synthesized compounds against hBuChE is stronger than that of the reference compound donepezil (hBuChE, IC 50 =2.13 μM), and the changes in compound chain length and substituents had little effect on the inhibitory activity of hBuChE, which was basically consistent with that of eqBuChE.

[0116] Experimental Example 2: Study on BACE-1 inhibitory activity of some target compounds

[0117] BACE-1 (Sigma) inhibition studies were performed using the peptide M-2420 (Bachem, Germany), which mimics the APP sequence, as a substrate. The experimental steps were as follows: 5 μL of the test compound (or DMSO in the control well) was preincubated with 175 μL of the enzyme (in 20 mM sodium acetate, pH 4.5, containing 0.1% CHAPS, w / v) at room temperature for 1 hour. The substrate (3 μM, final concentration) was then added and the reaction was allowed to proceed for 15 minutes. em =405nm(λ exc =320 nm). The DMSO concentration in the final mixture was kept below 5% (v / v) to prevent significant loss of enzyme activity. The background signal of the control wells (containing all reagents except BACE-1) was determined and subtracted. To demonstrate that BACE-1 activity was inhibited, a peptide-mimicking inhibitor (β-secretase inhibitor IV, Calbiochem, IC 5000) was serially diluted in the reaction wells. 50 =13.0±0.1 nM). The molar concentration of the compound that produces 50% inhibition was calculated using Graph Pad Prism version 7.00 software (GraphPad Software, San Diego, CA), which was the IC of the compound. 50 The results are expressed as the mean ± standard deviation (SD) of two independent experiments, each performed in triplicate.

[0118] Table 2. Inhibitory activity of some compounds against human BACE-1

[0119]

[0120] a The results are expressed as the mean (n=2) ± SD of two independent experiments.

[0121] Some compounds with good balanced inhibitory activity against AChE / BuChE were selected for hBACE-1 inhibitory activity study. As shown in Table 2, compound 7q (IC 50 =12.7μM) had the strongest inhibitory effect on BACE-1. In addition, the BACE-1 inhibitory activity of compounds with different linker lengths (7n~7p) was studied. Compounds containing 6 carbon spacers (7o, IC 50 =16.4 μM) than those containing 5 (7n, IC 50 =71.6 μM) and 8 (7p, IC 50 Compounds with a carbon interval greater than 100) have stronger BACE-1 inhibitory activity.

Claims

1. A cinnamic acid-capsaicin-tacrine derivative having the structure shown in the following formula 7 or a pharmaceutically acceptable salt thereof: ； in, R1 represents a hydrogen atom, a trifluoromethyl group, a methoxy group, a fluorine atom, a chlorine atom, or a bromine atom; R2 represents a hydrogen atom or a methoxy group; or R1 R2 represents a bis(methyleneoxy) group; R3 represents a hydrogen atom or a methoxy group; and n = 3 to 8.

2. The cinnamic acid-capsaicin-tacrine derivative according to claim 1, characterized in that: R1, R2 and R3 all represent hydrogen atoms, and n = 4, 5, 6, 7 or 8; R1 R2 represents a bis(methyleneoxy) group, R3 represents a hydrogen atom, and n = 5 or 6; R1 and R2 both represent methoxy groups, R3 represents a hydrogen atom, and n = 3, 5 or 6; R1, R2 and R3 all represent methoxy, n = 3, 4, 5, 6, 7 or 8; R1 represents a trifluoromethyl group, R2 and R3 both represent a hydrogen atom, and n = 3; R1 and R2 both represent fluorine atoms, R3 represents a hydrogen atom, and n = 3; R1 represents a chlorine atom, R2 and R3 both represent hydrogen atoms, and n = 3; R1 represents a bromine atom, R2 and R3 both represent hydrogen atoms, and n = 3.

3. The method for preparing the cinnamic acid-capsaicin-tacrine derivative according to claim 1, comprising the following steps: 1) placing a compound having a structure represented by the following formula 1 and a compound having a structure represented by the following formula 2 in an organic solvent and reacting them in the presence of a condensing agent and an alkaline reagent to obtain a compound having a structure represented by the following formula 5; wherein the condensing agent is a carbodiimide condensing agent or an organophosphorus condensing agent. When the condensing agent is a carbodiimide condensing agent, a condensation activator is also added; 2) taking the compound represented by the following formula 3 and the compound represented by the following formula 5, placing them in an organic solvent, adding an alkaline reagent to react, and obtaining a compound represented by the following formula 6; 3) taking the compound represented by the following formula 4 and the compound represented by the following formula 6, placing them in an organic solvent, adding an alkaline reagent to react, and obtaining the compound represented by the following formula 7; ； In the above formulae, R1 represents a hydrogen atom, a trifluoromethyl group, a methoxy group, a fluorine atom, a chlorine atom, or a bromine atom; R2 represents a hydrogen atom or a methoxy group; or R1 R2 represents a bis(methyleneoxy) group; R3 represents a hydrogen atom or a methoxy group; and n = 3 to 8.

4. The preparation method according to claim 3, wherein In the above steps, the organic solvent involved is one or a combination of two or more selected from dichloromethane, 1,2-dichloroethane, chloroform, chlorobenzene, N,N-dimethylformamide, dimethyl sulfoxide and acetonitrile; In the above steps, the alkaline reagent involved is one or a combination of two or more selected from sodium carbonate, potassium carbonate, cesium carbonate, potassium tert-butoxide, sodium tert-butoxide, sodium hydride, potassium hydride, triethylamine, sodium hydroxide and potassium hydroxide.

5. The preparation method according to claim 3 or 4, characterized in that: The method also includes the step of purifying the prepared compound having the structure shown in Formula 7.

6. Use of the cinnamic acid-capsaicin-tacrine derivative or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a medicament for treating or preventing Alzheimer's disease.

7. Use of the cinnamic acid-capsaicin-tacrine derivative or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a cholinesterase inhibitor or a BACE-1 inhibitor.

8. A pharmaceutical composition comprising a therapeutically effective dose of the cinnamic acid-capsaicin-tacrine derivative or a pharmaceutically acceptable salt thereof according to claim 1 as an active ingredient, and a pharmaceutically acceptable carrier.

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