Acetylcholinesterase and butyrylcholinesterase double-target inhibitor as well as preparation and application thereof

Abstract

The invention discloses an acetylcholin esterase and butyrylcholin esterase double-target inhibitor, which has the following chemical structural formula: wherein R is selected from any one of the following structural formulae;

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine synthesis, and particularly relates to a dual-target inhibitor of acetylcholinesterase and butyrylcholinesterase, and preparation and use thereof. BACKGROUND

[0002] Acetylcholinesterase and butyrylcholinesterase are two important cholinesterases in the human body. Acetylcholinesterase mainly exists in neuromuscular junctions, cholinergic synapses and red blood cells. Its core function is to hydrolyze the neurotransmitter acetylcholine very efficiently and specifically, making it rapidly inactivated, thereby precisely terminating nerve signal transmission and ensuring the normal operation of neuromuscular function.

[0003] Butyrylcholinesterase is widely distributed in plasma, liver, intestinal tract, brain glial cells and other parts. It has a wider range of hydrolysis substrates, and although it can also hydrolyze acetylcholine, it has higher hydrolysis efficiency for longer-chain choline esters such as butyrylcholine. It mainly participates in the metabolism of exogenous ester substances (such as certain drugs and toxins), and plays a role in the detoxification process.

[0004] Currently, there are cholinesterase inhibitor drugs in clinical use, such as galantamine, tacrine and rivastigmine, etc., for the prevention and treatment of Alzheimer's disease. However, long-term use of these drugs can cause obvious adverse reactions, including nausea, diarrhea, insomnia, vomiting, muscle spasms, fatigue, lethargy and loss of appetite, etc. The above drugs are all single-target designs for acetylcholinesterase, and have limited effect on Alzheimer's disease with complex etiology. Therefore, it is necessary to find a dual-target cholinesterase inhibitor with high efficiency and small side effects for the treatment of Alzheimer's disease.

[0005] Patent CN115925695B discloses a novel Stelleranoid A compound, its preparation method, pharmaceutical composition and application. The structure of Stelleranoid A is shown as formula 1

[0006] The Stelleranoid A disclosed in the application has good acetylcholinesterase and butyrylcholinesterase inhibitory activity, with IC50 values of 3.54±0.15 μM and 5.12±0.33 μM, close to the positive control drug galantamine (IC50 values of 15.81±0.41 μM and 55.62±0.27 μM). Patent CN102816107B discloses a carbazole derivative, its preparation method and use. The structure of 2-methoxy-5-[N-isopropyl-N-(3-nitropropionyl)] aminomethyl-9H-carbazole is shown as formula 2: The IC50 values are 2.2±0.5 μM and 2.5±0.9 μM, which are better than the positive control drug galantamine (19.6±8.1 μM and 80.1±11.1 μM).

[0007] Although the prior art performs relatively well in inhibiting acetylcholinesterase and butyrylcholinesterase activity, there is still room for further improvement. For example, the extraction conditions of Stelleranoid A in patent CN115925695B are complex, the yield is low, and the molecular weight > 500 is not conducive to the final drug, and the inhibitory activity needs to be further improved. The synthesis steps in patent CN102816107B are relatively cumbersome, the reaction conditions are relatively harsh, and the reagents used are relatively dangerous (such as: metallic sodium), making it difficult to implement large-scale production. Therefore, it is particularly important to develop new anti-Alzheimer's disease dual-target inhibitors, especially drugs with innovative skeletons. New inhibitors are expected to provide more effective treatment options and may provide new directions for the prevention and treatment of Alzheimer's disease in the future. SUMMARY

[0008] To solve the above technical problems, the present application provides an acetylcholinesterase and butyrylcholinesterase dual-target inhibitor and its preparation and use.

[0009] Specifically, the acetylcholinesterase and butyrylcholinesterase dual-target inhibitor has the structure shown in formula (I):

[0010] wherein the R group is selected from any one of the following structural formulae:

[0011]

[0012] Preferably, the chemical structure of the inhibitor is: and its IC50 values are 0.15±0.02 μM and 10.72±0.06 μM; It is worth mentioning that in this embodiment, the inhibitor has excellent acetylcholinesterase and butyrylcholinesterase inhibitory activity.

[0013] According to another aspect of the present application, the present application provides a preparation method of an acetylcholinesterase and butyrylcholinesterase dual-target inhibitor, which comprises:

[0014] S1001, forming a reaction system with cyclohexanone, sulfur powder and ethyl cyanoacetate, and after reaction, a first post-treatment is performed to obtain an intermediate of formula (II), i.e. ethyl 2-amino-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylate;

[0015] S1002, take the intermediate 2-amino-4,5,6,7-tetrahydrobenzothiophene-3-carboxylate obtained in step S1001, dissolve it in an organic solvent and form a reaction system with chloroacetyl chloride, and after the reaction, a second treatment is performed to obtain intermediate of formula (III), namely 2-(2-chloroacetamido)-4,5,6,7-tetrahydrobenzothiophene-3-carboxylate;

[0016] S1003, take the intermediate of formula (III) obtained in step S1002 and ammonia and dissolve them in an organic solvent to form a reaction system. After the reaction, the intermediate of formula (IV) is obtained by a third post-treatment, namely ethyl 2-(2-aminoacetamido)-4,5,6,7-tetrahydrobenzothiophene-3-carboxylate.

[0017] S1004: The intermediate of formula (IV) obtained in step S1003 and the substituted acyl chloride are dissolved in an organic solvent to form a reaction system. After the reaction, the inhibitor is obtained by the fourth post-treatment.

[0018] Preferably, in step S1001, at least one of the following conditions is satisfied:

[0019] Morpholine is used as the base, and ethanol is used as the organic solvent;

[0020] In step S1001, the reaction temperature is 44-55℃, preferably 50℃, and the reaction time is 4-6h, preferably 4h.

[0021] The ratio of cyclohexanone, sulfur powder, ethyl cyanoacetate, morpholine, and organic solvent added is (10~).

[0022] 20 mmol / L: (15-20) mmol / L: (15-20) mmol / L: (15-20) mL: (50-80) mL; preferably 10 mmol / L: 15 mmol / L: 20 mmol / L: 15 mmol / L: 60 mL.

[0023] Preferably, in step S1001,

[0024] The first post-treatment process is as follows: After the reaction is complete, the solvent is removed using a rotary evaporator to obtain the crude product. A small amount of solvent is added to dissolve the crude product, which is then transferred to a separatory funnel. The crude product is extracted sequentially with ethyl acetate and saturated NaCl solution, and this process is repeated three times. The organic phases are then combined. The organic layer is dried over anhydrous sodium sulfate, filtered, and purified by silica gel column chromatography to obtain intermediate (II).

[0025] Preferably, in step S1002, at least one of the following conditions is satisfied:

[0026] In step S1002, DMF is used as a catalyst, anhydrous K2CO3 is used as an acid-binding agent, and dichloromethane is used as the organic solvent.

[0027] In step S1002, the reaction temperature is 0-5°C, preferably 0°C, and the reaction time is 6-8h, preferably 6h;

[0028] The adding amount ratio of the intermediate of formula (II), chloroacetyl chloride, DMF, anhydrous K2CO3 and the organic solvent is (10-20) mmol:(15-20) mmol:(1-4) mmol:(20-40) mmol:(50-80) mL; preferably

[0029] 10 mmol: 15 mmol: 2 mmol: 30 mmol: 60 mL.

[0030] Preferably, in step S1002, the process of the second post-treatment is specifically as follows: after the reaction is completed, the reaction solution is poured into 50 mL of distilled water, and the water phase is extracted with 3x10 mL of dichloromethane three times, the organic phase is combined, and then washed with 0.5 mol / L hydrochloric acid solution, saturated sodium bicarbonate solution and saturated brine in sequence, the organic phase is dried with anhydrous sodium sulfate, and the organic phase solution is rotary evaporated into a solid state under reduced pressure, and then ultrasonic is applied until the system is uniform and there is no large blocky particles, to obtain the intermediate of formula (III).

[0031] Preferably, in the step S1003, at least one of the following conditions is met:

[0032] The organic solvent is methanol;

[0033] In step S1003, the reaction temperature is 55-65°C, preferably 55°C, and the reaction time is 10-14h, preferably 12h;

[0034] In step S1003, the adding amount ratio of the intermediate of formula (III), ammonia and the organic solvent is (10-20) mmol:(50-100) mmol:(10-20) mL. Preferably, it is 10 mmol: 60 mmol: 15 mL.

[0035] Preferably, in step S1003, the process of the third post-treatment is specifically as follows: after the material is completely converted, the reaction system is cooled to room temperature for filtration, the filter cake is reserved and washed with saturated brine to obtain a crude product, which is then subjected to column chromatography, dissolved in an eluent, rotary evaporated into a solid state under reduced pressure, and then ultrasonic is applied until the system is uniform and there is no large blocky particles, and then dried to obtain a pure compound of formula (IV) intermediate;

[0036] Preferably, DMF is used as the catalyst, anhydrous K2CO3 is used as the acid-binding agent, and dichloromethane is used as the organic solvent;

[0037] The temperature of the reaction in step S1004 is 0-15°C, preferably 5°C, and the reaction time is 8-10h, preferably 10h;

[0038] The adding amount ratio of the intermediate of formula (IV), different substituted acyl chloride, DMF, anhydrous K2CO3 and organic solvent in step S1004 is (10-20)mmol:(15-20)mmol:(1-4)mmol:(20-40)mmol:(50-80)ml; preferably 10mmol:10mmol:0.2mmol:15ml.

[0039] Preferably, in step S1004, the fourth post-treatment process is as follows: after the reaction is completed, the reaction solution is poured into 50ml of distilled water, 3x10ml of dichloromethane is added to extract the aqueous phase three times, the organic phase is combined, and the organic phase is sequentially washed with hydrochloric acid solution, saturated sodium bicarbonate solution and saturated brine, the organic phase is dried with anhydrous sodium sulfate, the solvent is removed under vacuum, and the pure compound is obtained by column chromatography.

[0040] According to another aspect of the present application, the present application provides a use of a acetylcholinesterase and butyrylcholinesterase dual-target inhibitor for preparing a drug capable of inhibiting acetylcholinesterase activity. BRIEF DESCRIPTION OF DRAWINGS

[0041] Figure 1 The preparation route of the acetylcholinesterase and butyrylcholinesterase dual-target inhibitor is shown; DETAILED DESCRIPTION

[0042] The preferred embodiments in the following description are only examples, and other obvious modifications can be conceived by those skilled in the art. The basic principles of the present application defined in the following description can be applied to other embodiments, modifications, improvements, equivalents and other technical solutions without departing from the spirit and scope of the present application.

[0043] A preparation method of an acetylcholinesterase and butyrylcholinesterase dual-target inhibitor according to a preferred embodiment of the present application is described in detail below, wherein the inhibitor has the following chemical structure formula:

[0044] Wherein, R is selected from any one of the following structural formulas:

[0045]

[0046] As preferred, the chemical structure formula of the acetylcholinesterase inhibitor is as follows:

[0047] The IC 50Values of 0.15 ± 0.02 μM and 10.72 ± 0.06 μM.

[0048] Reference Figure 1 The preparation method of the acetylcholinesterase and butyrylcholinesterase dual-target inhibitor comprises the following steps:

[0049] S1001, a reaction system is formed by cyclohexanone, sulfur powder and ethyl cyanoacetate, and after reaction, an intermediate of formula (II), i.e., ethyl 2-amino-4, 5, 6, 7-tetrahydrobenzothiophene-3-carboxylate, is obtained through first post-treatment.

[0050] Preferably, in step S1001, morpholine is used as the base, ethanol is used as the first organic solvent, the reaction temperature is 44-55°C, preferably 50°C, and the reaction time is 4-6h, preferably 4h. After reaction, the solvent is removed by a rotary evaporator, the obtained crude product is dissolved in a small amount of solvent, transferred to a separatory funnel, extracted with ethyl acetate and a saturated NaCl solution in sequence, and the organic phase is combined after repeated three times. The organic layer is dried over anhydrous sodium sulfate, filtered, and purified by silica gel column chromatography to obtain the intermediate of formula (II). The addition amount ratio of the cyclohexanone, sulfur powder, ethyl cyanoacetate, morpholine and organic solvent is (10-20) mmol:(15-20) mmol:(15-20) mmol:(15-20) mL:(50-80) mL.

[0051] Preferably, 10 mmol: 15 mmol: 20 mmol: 15 mmol: 60 mL.

[0052] Further, the preparation method further comprises:

[0053] Reference Figure 1 S1002, the intermediate ethyl 2-amino-4, 5, 6, 7-tetrahydrobenzothiophene-3-carboxylate obtained in step S1001 is dissolved in an organic solvent and chloroacetyl chloride to form a reaction system, and after reaction, an intermediate of formula (III), i.e., 2-ethoxycarbonyl-3-[(2-chloroacetyl)amino]-4, 5, 6, 7-tetrahydrobenzothiopyran-4-one, is obtained through second post-treatment.

[0054] Preferably, in step S1002, DMF is used as catalyst, anhydrous K2CO3 is used as acid-binding agent, the second organic solvent is dichloromethane, the reaction temperature is 0-5°C, preferably 0°C, and the reaction time is 6-8h, preferably 6h. The post-treatment process is as follows: after the reaction is completed, the reaction solution is poured into 50mL distilled water, 3x10mL dichloromethane is added to extract the aqueous phase three times, the organic phase is combined, and the organic phase is sequentially washed with 0.5moL / L hydrochloric acid solution, saturated sodium bicarbonate solution and saturated brine, the organic phase is dried with anhydrous sodium sulfate, and the organic phase solution is rotary evaporated into a solid under reduced pressure, ultrasonicated until the system is uniform and there are no large particles, to obtain the intermediate of formula (III). The addition amount ratio of the intermediate of formula (II), chloroacetyl chloride, DMF, anhydrous K2CO3 and the organic solvent is (10-20)mmoL:(15-20)mmoL:(1-4)mmoL:(20-40)mmoL:(50-80)mL.

[0055] Preferably, 10mmoL:15mmoL:2mmoL:30mmoL:60mL.

[0056] Further, the method further comprises:

[0057] Reference Figure 1 S1003, the intermediate of formula (III) obtained in step S1002 and ammonia are dissolved in an organic solvent to form a reaction system, and after reaction, the intermediate of formula (IV), i.e., 2-(2-aminoacetylamino)-4,5,6,7-tetrahydrobenzothiophene-3-carboxylic acid ethyl ester, is obtained through third post-treatment.

[0058] As a preferred, in step S1003, the third organic solvent is methanol, the reaction temperature is 55-65°C, preferably 55°C, and the reaction time is 10-14h, preferably 12h. The post-treatment process is as follows: the reaction temperature is 55-65°C, preferably 55°C, and the reaction time is 10-14h, preferably 12h. The addition amount ratio of the intermediate of formula (III), ammonia and the organic solvent is (10-20)mmoL:(50-100)mmoL:(10-20)mL.

[0059] Preferably, 10mmoL:60mmoL:15mL.

[0060] Further, the method further comprises:

[0061] Reference Figure 1 S1004, the intermediate of formula (IV) obtained in step S1003 and substituted acyl chloride are dissolved in an organic solvent to form a reaction system, and after reaction, the inhibitor is obtained through fourth post-treatment.

[0062] As preferred, DMF is used as catalyst, anhydrous K2CO3 is used as acid-binding agent, and dichloromethane is used as organic solvent. The reaction temperature is 0-15°C, preferably 5°C, and the reaction time is 8-10h, preferably 10h. The post-treatment process is as follows: after the reaction is completed, the reaction solution is poured into 50mL distilled water, 3x10mL dichloromethane is added to extract the aqueous phase three times, the organic phase is combined, and the organic phase is sequentially washed with hydrochloric acid solution, saturated sodium bicarbonate solution and saturated brine, the organic phase is dried with anhydrous sodium sulfate, the solvent is removed under vacuum, and the pure compound is obtained by column chromatography.

[0063] In step S1004, the adding amount ratio of the intermediate of formula (IV), different substituent acyl chloride, DMF, anhydrous K2CO3 and organic solvent is (10-20)mmoL:(15-20)mmoL:(1-4)mmoL:(20-40)mmoL:(50-80)mL. Preferably, 10mmoL:10mmoL:0.2mmoL:15mL.

[0064] The prepared inhibitor is subjected to acetylcholinesterase and butyrylcholinesterase inhibitor activity test, and the specific test method is as follows:

[0065] 1. Experimental instruments and materials

[0066] Multifunctional fluorescence enzyme marker, SP-Max 3500FL type, Shanghai Shanshu Biological Technology Co., Ltd.;

[0067] Super-clean workbench;

[0068] Bond A3Pipette manual single-channel adjustable pipette, 0.5-10ul, 10-100ul, 100-

[0069] 1000ul Titan Technology;

[0070] 96-well plate (white), sterilized, Corning;

[0071] Acetylcholinesterase and butyrylcholinesterase were purchased from Shanghai Nai Cheng Biological Technology Co., Ltd.; disodium hydrogen phosphate dodecahydrate, disodium hydrogen phosphate dihydrate, color developing agent 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and iodinated acetylthiocholine (ATCI) and iodinated butyrylthiocholine (BTCI) were purchased from Shanghai Titan Technology Co., Ltd.

[0072] Positive control drug, galantamine, purchased from Shanghai Titan Technology Co., Ltd.

[0073] 2. Reagent preparation

[0074] a. Phosphate buffer solution (PBS) 0.1 mol / L, pH = 7.60: 0.1 mM sodium dihydrogen phosphate solution mixed with 0.1 mM disodium hydrogen phosphate solution, adjust the pH of the solution to 7.60.

[0075] b. Substrate solution: first dissolve 72 mg ATCI in 10 mL PBS, then dilute 0.1 mL into 10 mL PBS to prepare a 0.25 mM solution, which can be used. BTCI is prepared in the same way.

[0076] c. Enzyme solution: weigh 25 mg of 200 U / g AChE and add 20 mL of PBS buffer to prepare a 0.25 U / mL stock solution, which can be used immediately, and is prepared immediately before use.

[0077] d. Color reagent: weigh 99 mg of DTNB dissolved in 10 mL of DMSO, take 0.1 mL and add it to 10 mL of DMSO to prepare a 0.25 mM solution, which can be used.

[0078] e. Positive control and sample solution: dissolve galantamine and synthetic compounds in DMSO, prepare an initial concentration of 1000 μmoL / L, and dilute it by a factor of 7 to prepare 7 concentration gradients, in the order of 500 μmoL / L, 100 μmoL / L, 10 μmoL / L, 1 μmoL / L, 0.1 μmoL / L, 0.05 μmoL / L, 0.005 μmoL / L, and prepare three groups for each concentration gradient.

[0079] Experimental method

[0080] Dissolve all test compounds and positive controls in DMSO and dilute to 8 different working concentrations. Then, add 40 μL of each concentration to a 96-well plate. At 37°C, add the same volume of AChE or BuChE solution to each well for pre-incubation for 30 minutes. After pre-incubation, add 40 μL of substrate, and finally add 40 μL of PBS solution to the 96-well plate. At the same time, establish a blank control group and add it to the PBS solution. Then add 40 μL of color reagent, incubate for 5 minutes, and use an enzyme marker to detect OD at 412 nm.

[0081] The experiment was divided into four groups, respectively

[0082] Sample group A (enzyme solution + substrate solution + buffer + test sample solution / galantamine solution + color reagent solution);

[0083] Sample background group A0 (substrate solution + buffer + test sample solution / galantamine solution + color reagent solution);

[0084] Negative control group B (enzyme solution + substrate solution + buffer + chromogenic agent solution + DMSO);

[0085] Negative background control group B0 (substrate solution + buffer + chromogenic agent solution + DMSO);

[0086] The amount of reagent added in each group is shown in Table 2. After measuring the absorbance value, the corresponding inhibition rate was calculated by the formula, and the curve and corresponding IC 50 value were fitted by Graphpad prism software. The inhibition rate of the sample on acetylcholinesterase was calculated as follows:

[0087]

[0088] Table 1: The amount of reagent added in four experimental groups

[0089]

[0090]

[0091] A represents the absorbance of the system in which the substrate, acetylcholinesterase / butyrylcholinesterase is incubated in the presence of sample solvent at 37℃ for 30 min.

[0092] A0 represents the background absorbance of the system in which acetylcholinesterase / butyrylcholinesterase is not added, and the sample and solvent are incubated for 30 min.

[0093] B represents the absorbance of the system in which the substrate, acetylcholinesterase / butyrylcholinesterase is incubated for 30 min.

[0094] B0 represents the absorbance of the system in which only the substrate and solvent are incubated for 30 min.

[0095] The present application will be described in detail below in conjunction with specific examples, but is by no means a limitation on the present application.

[0096] Example 1

[0097] 2-(2-(3,4,5-trimethoxybenzoylamino)acetylamino)-4,5,6,7-tetrahydrobenzothiophene-3-carboxylic acid ethyl ester, the structural formula of which is shown as follows:

[0098]

[0099] The specific synthesis steps are as follows:

[0100] (1) A round bottom flask was charged with cyclohexanone (0.491 g, 5 mmol), ethyl cyanoacetate (0.566 g, 5 mmol) and sulfur powder (0.160 g, 5 mmol). Anhydrous ethanol was used as solvent and morpholine as base. After the system was replaced with nitrogen, the reaction was heated under reflux at 50 °C in an oil bath. The progress of the reaction was monitored by thin layer chromatography. When the starting material spot disappeared, the reaction was terminated. The solvent was removed by rotary evaporation to obtain the crude product, which was dissolved in a small amount of solvent and transferred to a separatory funnel, which was extracted with EA and NaCl saturated solution, respectively. The organic phase was combined and dried over anhydrous sodium sulfate. The filtrate was purified by silica gel column chromatography. The solution was concentrated under reduced pressure to obtain a solid, which was sonicated until the system was uniform and free of large particles. The target product was isolated to obtain the intermediate of formula (II).

[0101] (2) A double-necked round bottom flask, constant pressure dropping funnel and thermometer were prepared and placed in an ice-salt bath. The intermediate of formula (II) (2.25 g, 10 mmol) was added, anhydrous dichloromethane was used as solvent, and the system was stirred and pre-cooled for 30 min. Then, 1.5 eq of potassium carbonate was added as an acid-binding agent, and a catalytic amount of DMF (0.1 mL) was added to promote the activation of the reaction. Chloroacetyl chloride (1.36 g, 12 mmol) was dissolved in pre-cooled anhydrous dichloromethane (10 mL) and transferred to the dropping funnel. Slowly added with a constant pressure dropping funnel, after the addition was completed, the reaction was carried out at room temperature for 8 h. When the reaction was terminated, the mixture was slowly poured into deionized water to quench the reaction, and the extraction was performed using a separatory funnel. The organic phase was combined and purified, the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography. The solution was concentrated under reduced pressure to obtain a solid, which was sonicated until the system was uniform and free of large particles. The target product was isolated to obtain the intermediate of formula (III).

[0102] (3) The intermediate of formula (III) (0.30 g, 1 mmol) and 30 mL of anhydrous methanol were added and magnetically stirred until completely dissolved. Concentrated ammonia (15 mL) was slowly added through a constant pressure funnel. The system was equipped with a reflux condenser under nitrogen protection, transferred to a pre-constant temperature oil bath, and the condensate was circulated for 10 h. The progress of the reaction was monitored by TLC sampling every 2 h. When the new product spot (Rf = 0.2) no longer changed, the reaction system was quickly cooled in an ice-water bath to terminate the reaction. The mixture was transferred to a separatory funnel and extracted after quenching. The organic phase was combined and purified, and column chromatography was performed using wet loading technique: the solution was concentrated under reduced pressure to obtain a solid, which was sonicated until the system was uniform and free of large particles. The target product was isolated by rotary evaporation to obtain the intermediate of formula (IV).

[0103] (4) A 100 mL dry two-necked round bottom flask was equipped with a magnetic stir bar, a thermometer and a constant pressure dropping funnel, and was pre-cooled in an ice-salt bath (-5-0 °C). The intermediate of formula (IV) (0.28 g, 1 mmol) and 15 mL of anhydrous dichloromethane were added and stirred uniformly. 3,4,5-Trimethoxybenzoyl chloride (0.18 mL, 1.2 mmol) was added dropwise, and after the addition, the reaction was continued at room temperature for 6 h. The reaction was monitored by TLC. After the reaction was completed, deionized water was slowly added to quench the reaction, and extraction was performed. The organic phase was combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. Column chromatography was performed, and the solution was concentrated under reduced pressure to obtain a solid. The solid was sonicated until it was uniform and free of large particles. The target product was separated by rotary evaporation to obtain the inhibitor of formula (I).

[0104] Experimental results

[0105] 2-(2-(3,4,5-Trimethoxybenzamido)acetamido)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid ethyl ester, white solid, yield 88%, inhibition IC50values: acetylcholinesterase: 0.15 ± 0.02 μM, butyrylcholinesterase: 10.72 ± 0.08 μM, and the corresponding IC50values of the positive control drug galantamine: acetylcholinesterase: 2.55 ± 0.01 μM, butyrylcholinesterase: 10.88 ± 0.03 μM.

[0106] 1 H NMR (401 MHz, Chloroform-d) δ 11.73 (s, 1H), 7.20 (d, J = 5.1 Hz, 1H), 7.18 (d, J = 3.4 Hz, 2H), 4.40 (d, J = 5.4 Hz, 2H), 4.31 (q, J = 7.1 Hz, 2H), 3.94 (s, 6H), 3.92 (s, 3H), 2.72 (dt, J = 49.2, 6.0 Hz, 4H), 1.81 (p, J = 6.6, 6.0 Hz, 4H), 1.38 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 167.68, 166.43, 166.28, 153.21, 153.21, 146.36, 141.13, 131.13, 129.05, 127.25, 112.49, 104.67, 104.67, 60.95, 60.64, 56.33, 56.33, 43.75, 24.42, 24.42, 22.97, 22.97, 22.97, 14.33. HRMS (ESI) calcd for C 23 H 28 N2O7S [M+Na]+: 499.1509; Found: 499.1501.

[0107] Example 2

[0108] 2-(2-(2-naphthoyl)acetamido)-4,5,6,7-tetrahydrobenzothiophene-3-carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a method similar to Example 1.

[0109]

[0110] White solid, yield 71%, IC50values for acetylcholinesterase: 2.22 ± 0.11 μΜ, butyrylcholinesterase: 11.94 ± 0.49 μΜ.

[0111] 1 H NMR (401 MHz, Chloroform-d) δ 11.75 (s, 1H), 8.39 (d, J = 7.5 Hz, 1H), 7.93 (d, J = 8.2 Hz, 1H), 7.85 (d, J = 6.9 Hz, 2H), 7.59 - 7.42 (m, 3H), 7.17 (s, 1H), 4.43 (d, J = 5.4 Hz, 2H), 4.35 - 4.22 (m, 2H), 2.85 - 2.58 (m, 4H), 1.80 (dt, J = 8.6, 5.5 Hz, 4H), 1.37 (s, 3H). 13 CNMR (101 MHz, Chloroform-d) δ 170.03, 166.37, 166.12, 146.57, 133.68, 133.28, 131.07, 131.00, 130.34, 128.24, 127.15, 127.09, 126.43, 125.81, 125.67, 124.70, 112.32, 60.59, 45.84, 26.36, 24.41, 22.99, 22.99, 14.32. HRMS (ESI) calcd for C 24 H 24 N2O4S [M+Na] + : 459.1348; Found: 459.1347.

[0112] Example 3

[0113] 2-(2-(3-cyclopentylpropanamido)acetamido)-4,5,6,7-tetrahydrobenzothiophene-3-carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a method similar to Example 1.

[0114]

[0115] Yellow solid, yield 77%, IC50values for acetylcholinesterase: 45.31 ± 0.03 μΜ, butyrylcholinesterase: 57.10 ± 0.11 μΜ.

[0116] 1 H NMR (401 MHz, Chloroform-d) δ 11.61 (s, 1H), 4.34 - 4.06 (m, 2H), 3.45 (d, J = 3.0 Hz, 2H), 3.13 - 3.06 (m, 4H), 2.67 (dt, J = 48.9, 6.3 Hz, 2H), 2.36 (d, J = 8.2 Hz, 1H), 2.03 (s, 2H), 1.74 (d, J = 9.2 Hz, 4H), 1.62 - 1.46 (m, 2H), 1.39 (td, J = 7.4, 1.8 Hz, 7H), 1.24 (t, J = 8.1 Hz, 1H), 1.10 (dd, J = 17.2, 8.7 Hz, 1H). 13 C NMR (101 MHz, Chloroform-d) δ 175.58, 170.39, 166.30, 146.50, 130.99, 126.94, 112.28, 60.51, 45.92, 43.22, 39.76, 32.46, 32.46, 31.64, 26.33, 25.17, 25.17, 24.39, 22.95, 22.79, 8.64. HRMS (ESI) calcd for C 21 H 30 N2O4S [M+Na] + : 429.1818; Found: 429.1826.

[0117] Example 4

[0118] 2-(2-(4-methoxybenzamide)acetylamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3- carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a similar method to Example 1.

[0119]

[0120] Yellow solid, yield 75%, IC50values for acetylcholinesterase: 3.11 ± 0.05 μΜ, butyrylcholinesterase: 77.95 ± 0.09 μΜ.

[0121] 1H NMR (401 MHz, Chloroform-d) δ 11.61 (s, 1H), 7.91 (d, J = 8.2 Hz, 2H), 7.60 (s, 1H), 6.90 (d, J = 7.9 Hz, 2H), 4.34 (s, 2H), 4.23 (q, J = 7.0 Hz, 2H), 3.83 (s, 3H), 2.66 (dt, J = 48.6, 5.8 Hz, 4H), 1.76 (d, J = 5.5 Hz, 4H), 1.31 - 1.27 (m, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 167.69, 166.73, 162.44, 146.26, 131.12, 131.12, 129.37, 126.94, 125.91, 125.91, 113.69, 113.69, 112.36, 60.54, 55.44, 43.81, 26.30, 24.38, 22.97, 22.79, 14.32. HRMS (ESI) calcd for C 21 H 24 N2O5S[M+H] + : 417.1478; Found: 417.1482.

[0122] Example 5

[0123] 2-(2-(4-(dimethylamino)benzamido)acetylamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3- carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a similar method to Example 1.

[0124]

[0125] White solid, yield 70%, IC50values: acetylcholinesterase: 2.80 ± 0.12 μΜ, butyrylcholinesterase: 70.23 ± 1.26 μΜ.

[0126] 1 H NMR (401 MHz, Chloroform-d) δ 11.61 (s, 1H), 7.91 (d, J = 8.2 Hz, 2H), 7.60 (s, 1H), 6.90 (d, J = 7.9 Hz, 2H), 4.34 (s, 2H), 4.23 (q, J = 7.0 Hz, 2H), 3.83 (s, 3H), 2.66 (dt, J = 48.6, 5.8 Hz, 4H), 1.76 (d, J = 5.5 Hz, 4H), 1.31 - 1.27 (m, 3H). 13C NMR (101 MHz, Chloroform-d) δ 174.01, 167.49, 165.86, 153.90, 151.63, 131.99, 129.20, 129.20, 127.03, 124.35, 123.00, 111.56, 111.54, 76.76, 60.46, 43.47, 41.64, 41.60, 24.39, 24.38, 22.99, 22.81, 14.24. HRMS (ESI) calcd for C 22 H 27 N3O4S[M+H] + : 430.1795; Found: 430.1796.

[0127] Example 6

[0128] 2-(2-(4-Fluorobenzamido)acetylamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3- carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a similar method to Example 1.

[0129]

[0130] White powdery solid, yield 71%, IC50values: acetylcholinesterase: 23.74 ± 1.50 μΜ, butyrylcholinesterase: 89.37 ± 2.61 μΜ.

[0131] 1 H NMR (401 MHz, Chloroform-d) δ 11.70 (s, 1H), 7.92 (dd, J = 8.8, 5.2 Hz, 2H), 7.25 (s, 1H), 7.12 (t, J = 8.6 Hz, 2H), 4.39 (d, J = 5.3 Hz, 2H), 4.30 (q, J = 7.1 Hz, 2H), 2.71 (dt, J = 47.9, 5.9 Hz, 4H), 1.81 (p, J = 6.7, 5.9 Hz, 4H), 1.37 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 166.94, 166.38, 166.11, 146.32, 131.11, 129.77, 129.77, 127.23, 115.74, 115.53, 115.53, 112.48, 60.66, 43.74, 26.35, 26.35, 22.80, 22.80, 14.30. HRMS (ESI) calcd for C 20 H 21 N2O4SF[M+Na] +: 427.1098; Found: 427.1101.

[0132] Example 7

[0133] 2-(2-(3,4,5-trifluorobenzamido)acetylamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3- carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a similar method to Example 1.

[0134]

[0135] White solid, yield 80%, IC50values for acetylcholinesterase: 37.82 ± 1.26 μΜ, butyrylcholinesterase: 15.37 ± 1.48 μΜ.

[0136] 1 H NMR (401 MHz, Chloroform-d) δ 11.77 (s, 1H), 7.61 (dd, J = 7.7, 6.3 Hz, 2H), 7.25 (s, 1H), 4.39 (d, J = 5.4 Hz, 2H), 4.33 (q, J = 7.1 Hz, 2H), 2.72 (dt, J = 46.0, 5.9 Hz, 4H), 1.81 (p, J = 6.1 Hz, 4H), 1.39 (t, J = 7.1 Hz, 3H).13C NMR (101 MHz, Chloroform-d) δ 172.08, 166.58, 165.54, 160.16, 149.33, 149.29, 146.24, 131.15, 127.41, 127.41, 112.31, 112.08, 112.08, 60.78, 43.74, 26.35, 26.35, 24.41, 24.41, 14.29. HRMS (ESI) calcd for C 20 H 19 N2O4SF3 [M+Na] + : 499.1509; Found: 499.1501.

[0137] Example 8

[0138] 2-(2-(4-nitrobenzamido)acetylamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a similar method to Example 1.

[0139]

[0140] White solid, yield 71%, IC50values for acetylcholinesterase: 40.23 ± 0.99 μΜ, butyrylcholinesterase: 109.38 ± 1.01 μΜ.

[0141] 1 H NMR (401 MHz, Chloroform-d) δ 11.74 (s, 1H), 8.11 (d, J = 8.1 Hz, 2H), 8.08 (s, 1H), 7.75 (d, J = 8.2 Hz, 2H), 4.39 (d, J = 5.5 Hz, 2H), 4.26 (q, J = 7.1 Hz, 2H), 2.70 (dt, J = 46.1, 5.8 Hz, 4H), 1.79 (q, J = 7.0, 6.5 Hz, 4H), 1.34 (t, J = 7.1 Hz, 3H). 13 CNMR (101 MHz, Chloroform-d) δ 171.85, 166.31, 166.01, 151.90, 146.30, 137.65, 132.32, 132.32, 131.14, 128.35, 128.35, 127.13, 123.29, 60.63, 45.91, 26.34, 26.33, 22.78, 22.78, 14.29. HRMS (ESI) calcd for C 20 H 21 N3O6S [M+Na] + : 454.1043; Found: 454.1041.

[0142] Example 9

[0143] 2-(2-(4-(Methoxycarbonyl)benzamido)acetylamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a method similar to Example 1.

[0144]

[0145] Yellow solid, yield 68%, IC50values for acetylcholinesterase: 17.37 ± 0.84 μΜ, butyrylcholinesterase: 13.63 ± 1.03 μΜ.

[0146] 1 H NMR (401 MHz, Chloroform-d) δ 11.74 (s, 1H), 8.11 (d, J = 8.1 Hz, 2H), 8.08 (s, 1H), 7.75 (d, J = 8.2 Hz, 2H), 4.39 (d, J = 5.5 Hz, 2H), 4.26 (q, J = 7.1 Hz, 2H), 2.70 (dt, J = 46.1, 5.8 Hz, 4H), 1.79 (q, J = 7.0, 6.5 Hz, 4H), 1.34 (t, J = 7.1 Hz, 3H). 13C NMR (101 MHz, Chloroform-d) δ 167.31, 166.38, 166.21, 166.17, 146.31, 137.65, 132.84, 131.10, 129.69, 129.69, 127.63, 127.63, 127.00, 112.40, 60.57, 45.89, 43.80, 26.31, 26.31, 22.96, 22.96, 14.29. HRMS (ESI) calcd for C 22 H 24 N2O6S[M+Na] + : 467.1247; Found: 467.1243.

[0147] Example 10

[0148] 2-(2-(4-cyanobenzamido)acetamido)-4,5,6,7-tetrahydrobenzo[b]thiophene-3- carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a similar method to Example 1.

[0149]

[0150] Yellow solid, yield 75%, IC50values for acetylcholinesterase: 30.62 ± 1.83 μΜ, butyrylcholinesterase: 23.69 ± 2.04 μΜ.

[0151] 1 H NMR (401 MHz, Chloroform-d) δ 11.76 (s, 1H), 8.29 (d, J = 9.0 Hz, 2H), 8.23 (s, 1H), 8.19 (d, J = 8.6 Hz, 2H), 4.41 (d, J = 5.6 Hz, 2H), 4.27 (q, J = 7.1 Hz, 2H), 2.69 (dd, J = 45.4, 5.7 Hz, 4H), 1.80 (p, J = 6.4 Hz, 4H), 1.34 (t, J = 7.2 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 171.77, 166.33, 165.98, 149.72, 139.36, 131.14, 131.14, 128.91, 128.91, 127.14, 126.65, 123.64, 119.11, 117.17, 60.63, 45.92, 26.34, 26.34, 24.42, 24.42, 14.28. HRMS (ESI) calcd for C 23 H 28 N2O7S[M+Na] +: 434.1144; Found: 434.1147.

[0152] Example 11

[0153] 2-(2-(5-chlorothiophene-2-carboxamido)acetylamino)-4,5,6,7-tetrahydrobenzo- thien-3-carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a similar method to Example 1.

[0154]

[0155] Yellow solid, yield 70%, IC50values for acetylcholinesterase: 13.25 ± 1.06 μΜ, butyrylcholinesterase: 63.25 ± 1.65 μΜ.

[0156] 1 H NMR (401 MHz, Chloroform-d) δ 11.70 (s, 1H), 7.96-7.88 (m, 3H), 7.24 (s, 1H), 4.39 (d, J = 5.3 Hz, 3H), 4.30 (q, J = 7.1 Hz, 3H), 2.81-2.61 (m, 6H), 1.81 (p, J = 6.7, 5.9 Hz, 6H), 1.37 (t, J = 7.1 Hz, 5H). 13 C NMR (101 MHz, Chloroform-d) δ 167.37, 165.85, 161.56, 146.14, 145.62, 136.77, 135.76, 131.14, 128.38, 127.24, 112.51, 60.68, 43.68, 26.32, 26.32, 22.98, 22.98, 14.35. HRMS (ESI) calcd for C 18 H 19 N2O4S2Cl [M+H]+: 427.0547; Found: 427.0539.

[0157] Example 12

[0158] 2-(2-cinnamoylaminoacetylamino)-4,5,6,7-tetrahydrobenzo-thien-3-carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a similar method to Example 1.

[0159]

[0160] White solid, yield 73%, IC50values for acetylcholinesterase: 65.78 ± 2.21 μΜ, butyrylcholinesterase: 111.21 ± 1.83 μΜ.

[0161] 1H NMR (401 MHz, Chloroform-d) δ 11.58 (s, 1H), 7.69 (d, J = 15.7 Hz, 1H), 7.53 (s, 2H), 7.36 (dt, J = 5.3, 2.8 Hz, 4H), 6.69 (d, J = 9.9 Hz, 1H), 4.29 (d, J = 7.9 Hz, 2H), 4.12 (d, J = 7.2 Hz, 2H), 2.69 (dt, J = 49.7, 5.7 Hz, 4H), 2.05 (s, 4H), 1.33 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 170.03, 166.77, 166.35, 146.30, 141.74, 131.16, 129.79, 129.79, 128.81, 128.81, 127.97, 127.97, 120.14, 112.41, 60.58, 45.91, 26.32, 24.40, 22.98, 21.07, 8.66. HRMS (ESI) calcd for C 22 H 24 N2O4S [M+Na] + : 435.1348; Found: 435.1341.

[0162] Example 13

[0163] 2-(2-(Furan-2-carboxamido)acetylamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3- carboxylic acid ethyl ester, whose structural formula is as follows, was prepared using a similar method to Example 1.

[0164]

[0165] White solid, yield 78%, IC50values for acetylcholinesterase: 20.33 ± 0.98 μΜ, butyrylcholinesterase: 58.86 ± 1.64 μΜ.

[0166] 1 H NMR (401 MHz, Chloroform-d) δ 11.58 (s, 1H), 7.69 (d, J = 15.7 Hz, 1H), 7.53 (s, 2H), 7.36 (dt, J = 5.3, 2.8 Hz, 4H), 6.69 (d, J = 9.9 Hz, 1H), 4.29 (d, J = 7.9 Hz, 2H), 4.12 (d, J = 7.2 Hz, 2H), 2.69 (dt, J = 49.7, 5.7 Hz, 4H), 2.05 (s, 4H), 1.33 (t, J = 7.1 Hz, 3H). 13C NMR (101 MHz, Chloroform-d) δ 166.16, 165.72, 158.67, 147.33, 146.27, 144.50, 131.16, 127.15, 116.41, 115.00, 112.18, 60.59, 45.85, 26.32, 26.32, 22.79, 22.79, 14.31. HRMS (ESI) calcd for C 18 H 20 N2O5S[M+Na] + : 399.0985; Found: 399.0990.

[0167] Example 14

[0168] 2-(2-(4-chlorophenoxy)acetamido)acetamido)-4,5,6,7-tetrahydrobenzo[b]thiophene-3- carboxylate, whose structural formula is as follows, was prepared using a method similar to Example 1.

[0169]

[0170] Yellow solid, yield 70%, IC50values for acetylcholinesterase: 44.24 ± 1.09 μΜ, butyrylcholinesterase: 121.19 ± 1.90 μΜ.

[0171] 1 H NMR (401 MHz, Chloroform-d) δ 11.69 (s, 1H), 7.41 (d, J = 11.3 Hz, 1H), 7.28 (d, J = 9.0 Hz, 2H), 7.02 - 6.77 (m, 2H), 4.61 (d, J = 5.7 Hz, 2H), 4.31 (t, J = 5.6 Hz, 2H), 3.62 - 3.39 (m, 2H), 2.71 (dd, J = 46.1, 6.7 Hz, 4H), 1.93 - 1.61 (m, 4H), 1.36 (q, J = 7.3, 5.5 Hz, 3H).13C NMR (101 MHz, Chloroform-d) δ 168.79, 166.57, 165.30, 155.85, 146.40, 131.08, 129.71, 129.71, 129.48, 127.31, 116.25, 116.25, 112.49, 67.69, 60.68, 42.74, 26.36, 24.42, 22.96, 22.79, 14.30. HRMS (ESI) calcd for C 21 H 23 N2O5SCl[M+H] +: 451.1088; Found: 451.1087.

[0172] Those skilled in the art will appreciate that the application described above is merely an example of the application. It is not intended to limit the application. The objects of the application have been fully and effectively achieved. The functional and structural principles of the application have been shown and described in the embodiments. Without departing from the principles described, the embodiments of the application can be modified or changed in any way.

Claims

1. A dual-target inhibitor of acetylcholinesterase and butyrylcholinesterase, characterized in that, The inhibitor has the following chemical structural formula: wherein R is selected from any one of the following structural formulas:

2. The acetylcholinesterase and butyrylcholinesterase dual target inhibitor according to claim 1, characterized in that, The chemical structural formula of the acetylcholinesterase inhibitor is as follows: Its IC 50 values are 0.15 ± 0.02 μΜ and 10.72 ± 0.06 μΜ.

3. A process for the preparation of a dual acetylcholinesterase and butyrylcholinesterase target inhibitor, characterized in that, The preparation method of the acetylcholinesterase and butyrylcholinesterase dual-target inhibitor comprises the following steps: S1001, three components of cyclohexanone, sulfur powder and ethyl cyanoacetate are formed into a reaction system, and after reaction, an intermediate of formula (II), i.e. ethyl 2-amino-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylate, is obtained through first post-treatment; S1002, the intermediate of ethyl 2-amino-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylate obtained in step S1001 is dissolved in an organic solvent and chloroacetyl chloride is formed into a reaction system, and after reaction, an intermediate of formula (III), i.e. ethyl 2-(2-chloroacetamido)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylate, is obtained through second post-treatment; S1003, the intermediate of formula (III) obtained in step S1002 and ammonia are dissolved in an organic solvent to form a reaction system, and after reaction, an intermediate of formula (IV), i.e. ethyl 2-(2-aminoacetamido)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylate, is obtained through third post-treatment; S1004, the intermediate of formula (IV) obtained in step S1003 and a substituted acyl chloride are dissolved in an organic solvent to form a reaction system, and after reaction, the inhibitor is obtained through fourth post-treatment.

4. The method of claim 3, wherein the acetylcholinesterase and butyrylcholinesterase dual target inhibitor is prepared by, In the step S1001, at least one of the following conditions is met: Morpholine is used as the base, and ethanol is used as the organic solvent; In step S1001, the reaction temperature is 44-55°C, preferably 50°C, and the reaction time is 4-6h, preferably 4h; The adding amount ratio of the cyclohexanone, sulfur powder, ethyl cyanoacetate, morpholine and organic solvent is (10-20)mmol:(15-20)mmol:(15-20)mmol:(15-20)mmol:(50-80)ml; preferably 10mmol:15mmol:20mmol:15mmol:60ml.

5. The method for preparing the dual-target inhibitor of acetylcholinesterase and butyrylcholinesterase according to claim 3, characterized in that, In the step S1001, The first post-treatment process is as follows: after the reaction is completed, the solvent is removed by a rotary evaporator to obtain a crude product, a small amount of solvent is added for dissolution, and then transferred to a separatory funnel, and then extracted with ethyl acetate and NaCl saturated solution in sequence, and the organic phase is combined after repeated three times. The organic layer is dried over anhydrous sodium sulfate, filtered, and purified by silica gel column chromatography to obtain the intermediate of formula (II).

6. The method of claim 3, wherein the acetylcholinesterase and butyrylcholinesterase dual target inhibitor is prepared by, In the step S1002, at least one of the following conditions is met: In step S1002, DMF is used as the catalyst, anhydrous K2CO3 is used as the acid-binding agent, and dichloromethane is used as the organic solvent; In step S1002, the reaction temperature is 0-5°C, preferably 0°C, and the reaction time is 6-8h, preferably 6h; The adding amount ratio of the intermediate of formula (II), chloroacetyl chloride, DMF, anhydrous K2CO3 and organic solvent is (10-20) mmol:(15-20) mmol:(1-4) mmol:(20-40) mmol:(50-80) mL; preferably 10 mmol:15 mmol:2 mmol:30 mmol:60 mL.

7. The method of claim 3, wherein the acetylcholinesterase and butyrylcholinesterase dual target inhibitor is prepared by, In step S1002, after the reaction is completed, the reaction solution is poured into 50 mL of distilled water, 3x10 mL of dichloromethane is added to extract the water phase three times, the organic phases are combined, and the organic phase is washed with 0.5 mol / L hydrochloric acid solution, saturated sodium bicarbonate solution and saturated brine in sequence, the organic phase is dried with anhydrous sodium sulfate, the organic phase solution is rotary evaporated into a solid state under reduced pressure, and the system is ultrasonically treated until it is uniform and there are no large blocky particles, to obtain the intermediate of formula (III).

8. The method of claim 3, wherein the acetylcholinesterase and butyrylcholinesterase dual target inhibitor is prepared by, In step S1003, at least one of the following conditions is met: The organic solvent is methanol; In step S1003, the reaction temperature is 55-65℃, preferably 55℃, and the reaction time is 10-14 h, preferably 12 h; In step S1003, the adding amount ratio of the intermediate of formula (III), ammonia and organic solvent is (10-20) mmol:(50-100) mmol:(10-20) mL; preferably 10 mmol:60 mmol:15 mL.

9. The preparation method of the acetylcholinesterase and butyrylcholinesterase dual-target inhibitor according to claim 3, characterized in that, In step S1003, after the material is completely converted, the reaction system is cooled to room temperature for filtration, the filter cake is reserved and washed with saturated brine to obtain a crude product, which is subjected to column chromatography separation, dissolved in an eluent, rotary evaporated into a solid state under reduced pressure, and ultrasonically treated until the system is uniform and there are no large blocky particles, and then dried to obtain the pure compound intermediate of formula (IV); 10. The preparation method of the acetylcholinesterase and butyrylcholinesterase dual-target inhibitor according to claim 3, characterized in that, DMF is used as a catalyst, anhydrous K2CO3 is used as an acid-binding agent, and dichloromethane is used as the organic solvent; In step S1004, the reaction temperature is 0-15℃, preferably 5℃, and the reaction time is 8-10 h, preferably 10 h; In step S1004, the adding amount ratio of the intermediate of formula (IV), different substituent acyl chloride, DMF, anhydrous K2CO3 and organic solvent is (10-20) mmol:(15-20) mmol:(1-4) mmol:(20-40) mmol:(50-80) mL; preferably 10 mmol:10 mmol:0.2 mmol:15 mL.

11. The preparation method of the acetylcholinesterase and butyrylcholinesterase dual-target inhibitor according to claim 3, characterized in that, In step S1004, the fourth post-processing process is specifically as follows: after the reaction is completed, the reaction solution is poured into 50 mL of distilled water, the water phase is extracted with 3×10 mL of dichloromethane three times, the organic phases are combined, the organic phase is sequentially washed with hydrochloric acid solution, saturated sodium bicarbonate solution and saturated brine, the organic phase is dried with anhydrous sodium sulfate, the solvent is removed in vacuum, and the pure compound is obtained by column chromatography to obtain the inhibitor shown in formula (I).

12. Use of a dual butyrylcholinesterase and acetylcholinesterase target inhibitor according to any one of claims 1 to 11 for the manufacture of a medicament capable of inhibiting acetylcholinesterase activity.

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