A pyrazolone-tacrine compound and its preparation method and application

Abstract

The present invention discloses a pyrazolone-tacrine compound and its preparation method and application. The compound is a compound represented by formula (I), an enol tautomer, a polymorph, a solvate or a pharmaceutically acceptable salt thereof. The beneficial effects of the present invention are as follows: the present invention provides a series of pyrazolone-tacrine compounds with novel structures, which can exert excellent neuroprotective effects from multiple aspects such as inhibiting acetylcholinesterase activity, resisting oxidative stress and chelating toxic metal ions, and play an important role in preventing or treating neurodegenerative diseases caused by damage to the cholinergic system, such as Alzheimer's disease and its complications.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to a pyrazolone-tacrine compound and a preparation method and application thereof. Background Art

[0002] Neurodegenerative diseases are caused by the loss of neurons and / or their myelin sheaths, which deteriorate over time and cause functional impairment. They can be divided into acute neurodegenerative diseases and chronic neurodegenerative diseases. The former mainly includes cerebral ischemia (CI), brain injury (BI), and epilepsy; the latter includes Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), different types of spinocerebellar ataxia (SCA), Pick's disease, etc. For neurodegenerative diseases, existing therapeutic drugs mainly include acetylcholinesterase inhibitors and free radical scavengers.

[0003] Acetylcholine is a neurotransmitter that is believed to be related to various behaviors such as learning and memory. Acetylcholine will be hydrolyzed by acetylcholinesterase, causing a decrease in acetylcholine levels in the brain, which in turn leads to a decrease in learning, memory, and action abilities. If not intervened, it will lead to the occurrence of neurodegenerative diseases such as Alzheimer's disease (AD). The tetrahydroacridine drug tacrine is a first-generation reversible anticholinesterase inhibitor. It is highly lipid-soluble and can easily pass through the blood-brain barrier. It can not only inhibit the degradation of acetylcholine by acetylcholinesterase and increase the content of acetylcholine in the body, but also excite M and N receptors, increasing the release of acetylcholine in cognitive function-related areas of the brain.

[0004] On the other hand, normal cell aerobic metabolism will produce oxygen free radicals and nitrogen free radicals, including superoxide anion free radicals, nitric oxide free radicals, etc. Among them, reactive oxygen free radicals (ROS) will cause a series of biochemical reactions, destroy the cell structure, cause cell necrosis, and cause various diseases. The brain is one of the organs with the highest oxygen-sugar ratio and the most active metabolism in the body. Therefore, some cells in brain tissue, such as cholinergic neurons and glial cells, are more likely to produce free radicals, which will cause irreversible damage to neuronal cells and lead to neurodegenerative diseases. The pyrazolone compound Edaravone is a free radical scavenger that can scavenge free radicals by providing electrons, thereby inhibiting lipid peroxidation reactions, reducing the damage of free radicals to cells, and protecting nerve cells.

[0005] All of these existing drugs are expected to achieve the therapeutic effect of neurodegenerative diseases through a certain pathway of action. However, given the diversity of pathogenic factors of neurodegenerative diseases, a single pathway of action cannot significantly reduce the overall dysfunction and loss of neurons, which also makes the current drug treatment limited and unable to cope with advanced neurodegenerative diseases. Therefore, it is very necessary to develop a drug that improves neurodegenerative diseases through multiple pathways to further improve the treatment effect. Summary of the invention

[0006] In view of the shortcomings of the prior art, the present invention provides a pyrazolone-tacrine compound and its preparation method and application. The compound can exert neuroprotective effects in multiple ways and is expected to be developed into a new drug for treating neurodegenerative diseases.

[0007] In a first aspect of the present invention, a pyrazolone-tacrine compound is provided, which is a compound represented by formula (I), an enol tautomer, a polymorph, a solvate or a pharmaceutically acceptable salt thereof;

[0008]

[0009] Wherein, Z is selected from C6-C10 aromatic hydrocarbon groups, substituted or unsubstituted 5-7 membered aromatic heterocyclic groups, substitution means substitution by 1-3 substituents selected from halogen atoms and haloalkyl groups; X is hydrogen or a halogen atom; n is an integer of 1-3; m is an integer of 1-2.

[0010] According to a specific embodiment of the present invention, Z is selected from phenyl, a substituted or unsubstituted 6-membered aromatic heterocyclic group, and the heteroatom in the aromatic heterocyclic group is a nitrogen atom.

[0011] According to a specific embodiment of the present invention, Z is selected from any one of the following:

[0012] According to a specific embodiment of the present invention, the compound represented by formula (I) is selected from any one of the following ET1 to ET21:

[0013]

[0014]

[0015]

[0016] The second aspect of the present invention provides a method for preparing the aforementioned pyrazolone-tacrine compound, comprising: subjecting the compound of formula (1) to a cyclization reaction with dimethyl butynedioate to obtain a compound of formula (2); subjecting the compound of formula (2) to a hydrolysis reaction under alkaline conditions to obtain a compound of formula (3); subjecting the compound of formula (3) to a condensation reaction with a compound of formula (4) to obtain a compound of formula (I);

[0017]

[0018] Wherein, the definitions of Z, X, n, and m are as described above.

[0019] According to a specific embodiment of the present invention, the molar ratio of the compound represented by formula (1) to dimethyl butynedioate is 1:0.9-1.5, and the conditions for the cyclization reaction include: in the presence of methanol, reacting at 0°C for 2-4 hours, adding sodium methoxide to the reaction solution, and refluxing at 60-80°C for 0.5-2 hours.

[0020] According to a specific embodiment of the present invention, the conditions for the hydrolysis reaction include: reflux at 30-50° C. for 4-6 hours in the presence of methanol, tetrahydrofuran and a base.

[0021] According to a specific embodiment of the present invention, the molar ratio of the compound represented by formula (3) to the compound represented by formula (4) is 1:1.0-1.5, and the conditions of the condensation reaction include: reacting at room temperature for 20-30 hours under the protection of an inert atmosphere and in the presence of a solvent and a condensation agent.

[0022] The third aspect of the present invention provides a pharmaceutical composition comprising the aforementioned pyrazolone-tacrine compound; preferably, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients; preferably, the pharmaceutical composition is in the form of capsules, pills, tablets, granules or injections.

[0023] The fourth aspect of the present invention provides the use of the aforementioned pyrazolone-tacrine compounds in any of the following i) to v): i) for preparing drugs for inhibiting acetylcholinesterase activity; ii) for preparing drugs for scavenging free radicals; iii) for preparing drugs for protecting neuronal cells; iv) for preparing drugs for improving animal memory and cognition; v) for preparing drugs for preventing, treating or assisting in the treatment of neurodegenerative diseases.

[0024] The beneficial effects of the present invention are:

[0025] The present invention provides a series of pyrazolone-tacrine compounds with novel structures, which can exert excellent neuroprotective effects from multiple aspects such as inhibiting acetylcholinesterase activity, resisting oxidative stress and chelating toxic metal ions, and play an important role in preventing or treating neurodegenerative diseases caused by damage to the cholinergic system, such as Alzheimer's disease and its complications. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a synthetic route for the pyrazolone-tacrine compounds of the present invention;

[0027] Figure 2 Characterization diagram of the chelating ability of the target compound with different metals;

[0028] Figure 3 This is a behavioral characterization diagram of the target compound on ICR mice;

[0029] Figure 4 This is the Nissl staining image of the hippocampus of ICR mice;

[0030] Figure 5 These are glycine silver staining images of the hippocampus and H&E staining images of the liver of ICR mice. DETAILED DESCRIPTION

[0031] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention is clearly and completely described below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0032] The solvents used in the examples of the present invention are all commercially available chemically pure or analytically pure. The structures of the compounds were determined by nuclear magnetic resonance (NMR). The NMR was determined using a Bruker AVANCE-600 NMR spectrometer, the solvents used were CDCl3 or DMSO-d6, and the internal standard was TMS.

[0033] The method for preparing pyrazolone-tacrine compounds of the present invention comprises the following steps:

[0034] (1) Compound 1 is dissolved in methanol to prepare a solution with a molar concentration of 0.85-1.5 mol / L, and a methanol solution containing 0.9-1.5 eq of dimethyl butynedioate is added dropwise under ice bath conditions. The reaction solution is reacted at 0° C. for 2-4 hours, and then 1.0-2.0 eq of sodium methoxide is added to the reaction solution. The reaction solution is heated under reflux at 60-80° C. for 0.5-2 hours to obtain compound 2;

[0035] (2) Compound 2 was dissolved in a mixed solution of methanol and tetrahydrofuran (THF) in a volume ratio of 4:3 to prepare a solution with a molar concentration of 0.3 to 0.6 mol / L, and 1.5 to 2.5 eq of a 2 mol / L sodium hydroxide aqueous solution was added dropwise under an ice bath to react and obtain Compound 3;

[0036] (3) Compound 3 is dissolved in N,N-dimethylformamide (DMF) to prepare a solution with a molar concentration of 0.05-0.1 mol / L, and then 2.0-3.5 eq of N,N-diisopropylethylamine (DIPEA) and 1.0-1.5 eq of condensing agent O-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU) are added to the reaction solution, and finally 1.0-1.5 eq of the corresponding amine compound is added to the reaction solution to prepare pyrazolone-tacrine compound ET1-18 (I); or,

[0037] Compound 3 was dissolved in dry dichloromethane (DCM) to prepare a solution with a molar concentration of 0.05-0.1 mol / L, and 0.8-1.2 eq of 1-ethyl-3 (3-dimethylpropylamine) carbodiimide (EDCI) and 1-hydroxybenzotriazole (HOBT) were added to the reaction solution, and finally 1.0-1.5 eq of the corresponding amine compound was added to the reaction solution to prepare pyrazolone-tacrine compound ET19-21 (I);

[0038] Among them, amine compounds include N 1 -(1,2,3,4-tetrahydroacridin-9-yl)ethane-1,2-diamine (8a), N 1 -(1,2,3,4-tetrahydroacridin-9-yl)propane-1,3-diamine (8b), N 1 -(6-Chloro-1,2,3,4-tetrahydroacridin-9-yl)ethane-1,2-diamine (8c), N 1 -(6-chloro-1,2,3,4-tetrahydroacridin-9-yl)propane-1,3-diamine (8d), N 1 -(6-Chloro-1,2,3,4-tetrahydroacridin-9-yl)butane-1,4-diamine (8e), N 1 -(6-chloro-2,3-dihydro-1H-cyclopenta[b]quinolin-9-yl)propane-1,3-diamine (8f), N 1 -(2,3-dihydro-1H-cyclopenta[b]quinolin-9-yl)ethane-1,2-diamine (8 g), N 1 -(2,3-Dihydro-1H-cyclopenta[b]quinolin-9-yl)propane-1,3-diamine (8h).

[0039] The synthetic route of the pyrazolone-tacrine compounds of the present invention is as follows Figure 1 As shown, wherein Z is selected from C6-C10 aromatic hydrocarbon groups, substituted or unsubstituted 5-7 membered aromatic heterocyclic groups, substituted by 1-3 substituents selected from halogen atoms and haloalkyl groups, preferably Z is phenyl, substituted or unsubstituted 6 membered aromatic heterocyclic groups, the hetero atom in the aromatic heterocyclic group is a nitrogen atom, more preferably: X is a hydrogen or halogen atom; n is an integer of 1 to 3; and m is an integer of 1 to 2.

[0040] The preparation method of the above-mentioned pyrazolone-tacrine compounds is exemplified below in conjunction with specific examples.

[0041] Example 1: Preparation of compound 5-hydroxy-1-phenyl-1H-pyrazole-3-carboxylic acid methyl ester (2a)

[0042]

[0043] Compound phenylhydrazine (1a, 12g, 110mmol) was dissolved in methanol (120mL), and a methanol (80mL) solution containing dimethyl butynedioate (15.2mL, 124mmol) was added dropwise under ice bath conditions. After the reaction solution was reacted at 0℃ for 3 hours, sodium methoxide (9.2g, 172mmol) was added to the reaction solution, and refluxed at 70℃ for 1 hour. After the reaction was completed, the methanol solvent was concentrated to remove it, 80mL of water was added, and 2mol / LHCl was added dropwise under ice bath conditions until the pH of the reaction solution was 4, and a yellow precipitate was precipitated. It was filtered by suction, and the filter cake was washed with water and dried to obtain a yellow powder 2a with a yield of 60%.

[0044] Example 2: Preparation of compound 5-hydroxy-1-phenylpyrazole-3-carboxylic acid (3a)

[0045]

[0046] Compound 2a (17.4 g, 80 mmol) was dissolved in a mixture of methanol (120 mL) and tetrahydrofuran (90 mL). 2 mol / L NaOH (90 mL, 180 mmol) was added dropwise under ice bath, and the reaction solution was refluxed at 40°C for 5 hours. After the reaction was completed, 2 equivalents of water were added, the reaction solution was concentrated to remove methanol, and 2 mol / L HCl was added dropwise under ice bath until the pH of the reaction solution was 4. A large amount of light yellow solid was precipitated, which was filtered by suction, and the filter cake was washed with water and dried to obtain light yellow solid 3a with a yield of 65%.

[0047] Example 3: Preparation of compound 5-hydroxy-1-pyridin-2-ylpyrazole-3-carboxylic acid (3b)

[0048]

[0049] Intermediate 3b is prepared by replacing phenylhydrazine (1a) with 2-hydrazinepyridine (1b) and reacting with dimethyl butynedioate, followed by hydrolysis. The reaction process is similar to 3a, and the product is a yellow powder 3b with a yield of 55%. 1 HNMR (300MHz, DMSO-d6) δ8.53 (ddd, J=5.0, 1.9, 0.9Hz, 1H), 8.09 (ddd, J=8.3, 7.5, 1. 9Hz, 1H), 7.81 (dt, J=8.3, 1.0Hz, 1H), 7.46 (ddd, J=7.5, 5.0, 1.1Hz, 1H), 5.99 (s, 1H).

[0050] Example 4: Preparation of compound 1-(5-fluoropyridin-2-yl)-5-hydroxy-1H-pyrazole-3-carboxylic acid (3c)

[0051]

[0052] Intermediate 3c is prepared by replacing phenylhydrazine (1a) with 5-fluoro-2-hydrazinepyridine (1c) and reacting with dimethyl butynedioate, followed by hydrolysis. The reaction process is similar to 3a, and the product is a yellow powder 3c with a yield of 58%. Melting point: 229.3℃–232.7℃. 1 HNMR (600MHz, DMSO-d6) δ8.56(d,J=3.0Hz,1H),7.99(td,J=8.6,3.1Hz,1H),7.80(dd,J=9.0,3.9Hz,1H),5.95(s,1H).

[0053] Example 5: Preparation of compound 1-(5-chloropyridin-2-yl)-5-hydroxy-1H-pyrazole-3-carboxylic acid (3d)

[0054]

[0055] Intermediate 3d is prepared by replacing phenylhydrazine (1a) with 5-chloro-2-hydrazinepyridine (1d) and reacting with dimethyl butynedioate, followed by hydrolysis. The reaction process is similar to 3a, and the product is a dark yellow solid 3d with a yield of 70%. Melting point: 216.2℃–220.5℃. 1 HNMR (600MHz, DMSO-d6) δ12.14(s,1H),8.60(d,J=2.5Hz,1H),8.16(dd,J=8.7,2.6Hz,1H),7.80(d,J=8.7Hz,1H),5.98(s,1H).

[0056] Example 6: Preparation of compound 5-hydroxy-1-(pyrimidin-2-yl)-1H-pyrazole-3-carboxylic acid (3e)

[0057]

[0058] Intermediate 3e is prepared by replacing phenylhydrazine (1a) with 2-hydrazinepyrimidine (1e) and reacting with dimethyl butynedioate, followed by hydrolysis. The reaction process is similar to 3a, and the product is an orange flaky solid 3e with a yield of 63%. Melting point: 162.4℃–172.9℃. 1 HNMR (600MHz, DMSO-d6) δ12.94(s,1H),12.02(s,1H),8.95(d,J=4.9Hz,2H),7.60(t,J=4.9Hz,1H),5.98(s,1H).

[0059] Example 7: Preparation of compound 5-hydroxy-1-(5-(trifluoromethyl)pyridin-2-yl)-1H-pyrazole-3-carboxylic acid (3f)

[0060]

[0061] Intermediate 3f is prepared by replacing phenylhydrazine (1a) with 5-trifluoromethyl-2-hydrazinepyridine (1f) and reacting with dimethyl butynedioate, followed by hydrolysis. The reaction process is similar to 3a, and the product is an orange-red powder 3f with a yield of 65%. Melting point: 137.4℃–144.5℃. 1 HNMR (600MHz, DMSO-d6) δ13.05(s,1H),12.31(s,1H),8.95(dt,J=2.8,1.0Hz,1H),8.44(dd,J=8.8,2.5Hz,1H),8.02(d,J=8.6Hz,1H),6.01(s,1H).

[0062] Example 8: Preparation of compound 1-(3-chloropyridin-2-yl)-5-hydroxy-1H-pyrazole-3-carboxylic acid (3 g)

[0063]

[0064] Intermediate 3g is prepared by replacing phenylhydrazine (1a) with 3-chloro-2-hydrazinopyridine (1g) and reacting with dimethyl butynedioate, followed by hydrolysis. The reaction process is similar to 3a, and the product is a gray powder of 3g with a yield of 60%.

[0065] Example 9: Preparation of Compound 9-Chloro-1,2,3,4-tetrahydroacridine (7a)

[0066]

[0067] POCl3 (90 mL) was added dropwise to a flask containing anthranilic acid (11.1 g, 81 mmol) and cyclohexanone (11.7 mL, 113 mmol) at low temperature. The reaction solution was refluxed at 120 °C for 5 h, cooled, concentrated to remove POCl3, and an appropriate amount of water was added.

[0068] Saturated Na2CO3 solution was added dropwise at low temperature to precipitate a light yellow precipitate, which was filtered and dried to obtain a yellow powder 7a.

[0069] Example 10: Preparation of compound 6,9-dichloro-1,2,3,4-tetrahydroacridine (7b)

[0070]

[0071] Intermediate 7b is obtained by replacing anthranilic acid with 4-chloro-2-aminobenzoic acid and refluxing with cyclohexanone in the presence of POCl3. The reaction process is similar to that of 7a. The product is a brown powder.

[0072] Example 11: Preparation of Compound 9-chloro-2,3-dihydro-1H-cyclopenta[b]quinoline (7c)

[0073]

[0074] Intermediate 7c is obtained by replacing the cyclohexanone in Example 9 with cyclopentanone and refluxing in the presence of POCl3, and the reaction process is similar to that of 7a. The product is a black powder.

[0075] Example 12: Preparation of Compound 6,9-Dichloro-2,3-dihydro-1H-cyclopenta[b]quinoline (7d)

[0076]

[0077] Intermediate 7d is obtained by replacing the anthranilic acid in Example 9 with 4-chloro-2-aminobenzoic acid and refluxing with cyclopentanone in the presence of POCl3. The reaction process is similar to that of 7a. The product is a brown powder.

[0078] Example 13: Preparation of Compound N 1 -(1,2,3,4-Tetrahydroacridin-9-yl)ethane-1,2-diamine (8a)

[0079]

[0080] Take ethylenediamine hydrochloride (15g, 115mmol) in a flask, add methanol to dissolve, add methanol (12g, 230mmol) dissolved in NaOCH3 dropwise in an ice bath, and react in an ice bath for 1h. Remove methanol, add 7a (5g, 23mmol) in a flask, add phenol (solid, liquid when heated and stirred) (15g), add KI (15mg), and react at 169℃ under N2 protection for 6h. After cooling, add KOH solution (15mol / L) dropwise under stirring, adjust the pH to alkaline, extract with DCM, concentrate the organic phase, and purify by silica gel column chromatography (DCM:MeOH=10:1, v / v) to obtain compound 8a.

[0081] Example 14: Preparation of Compound N 1 -(1,2,3,4-Tetrahydroacridin-9-yl)propane-1,3-diamine (8b)

[0082]

[0083] Compound 7a reacts with 1,3-propylenediamine in the presence of phenol as a solvent. The reaction process is similar to that of compound 8a. Compound 8b is obtained by purification by silica gel column chromatography.

[0084] Example 15: Preparation of Compound N 1 -(6-Chloro-1,2,3,4-tetrahydroacridin-9-yl)1,2-diamine (8c)

[0085]

[0086] Compound 7b was reacted with ethylenediamine in the presence of isoamyl alcohol as solvent. The reaction process was similar to that of compound 8a. Compound 8c was obtained by purification by silica gel column chromatography (DCM:MeOH=8:1, v / v).

[0087] Example 16: Preparation of Compound N 1 -(6-Chloro-1,2,3,4-tetrahydroacridin-9-yl)propane-1,3-diamine (8d)

[0088]

[0089] Compound 7b was reacted with 1,3-propylenediamine in the presence of isopentanol as solvent. The reaction process was similar to that of compound 8a. Compound 8d was obtained by purification by silica gel column chromatography (DCM:MeOH=8:1, v / v).

[0090] Example 17: Preparation of Compound N 1 -(6-Chloro-1,2,3,4-tetrahydroacridin-9-yl)butane-1,4-diamine (8e)

[0091]

[0092] Compound 7b reacts with 1,4-butanediamine in isopentanol as solvent. The reaction process is similar to that of compound 8a. Compound 8e is obtained by purification by silica gel column chromatography.

[0093] Example 18: Preparation of Compound N 1 -(6-Chloro-2,3-dihydro-1H-cyclopenta[b]quinolin-9-yl)propane-1,3-diamine (8f)

[0094]

[0095] Compound 7d reacts with 1,3-propylenediamine in isopentanol as solvent. The reaction process is similar to that of compound 8a. Compound 8f is obtained by purification by silica gel column chromatography.

[0096] Example 19: Preparation of Compound N 1 -(2,3-Dihydro-1H-cyclopenta[b]quinolin-9-yl)ethane-1,2-diamine (8 g)

[0097]

[0098] Compound 7c was reacted with ethylenediamine in the presence of phenol as solvent. The reaction process was similar to that of compound 8a. Compound 8g was obtained by purification by silica gel column chromatography (DCM:MeOH=8:1, v / v).

[0099] Example 20: Preparation of Compound N 1 -(2,3-Dihydro-1H-cyclopenta[b]quinolin-9-yl)propane-1,3-diamine (8h)

[0100]

[0101] Compound 7c was reacted with 1,3-propylenediamine in the presence of phenol as solvent. The reaction process was similar to that of compound 8a. Compound 8h was obtained by purification via silica gel column chromatography (DCM:MeOH=8:1, v / v).

[0102] Example 21: Preparation of compound 5-hydroxy-1-(pyridin-2-yl)-N-(2-((1,2,3,4-tetrahydroacridin-9-yl)amino)ethyl)-1H-pyrazole-3-carboxamide (ET1)

[0103]

[0104] Compound 3b (205 mg, 1 mmol) was dissolved in DMF (15 mL), and DIPEA (176 μL, 1 mmol) was added. HATU (448 mg, 1.17 mmol) was added under ice bath, and compound 8a (241 mg, 1 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. Insoluble matter was precipitated after adding water (30 mL), which was filtered, and the filter cake was washed with water and dried. The target compound ET1 was purified by silica gel column chromatography as a yellow powder (200 mg) with a yield of 46%. Melting point: 214°C–217°C. 1 H NMR (400MHz, DMSO-d6) δ8.61(t,J=6.0Hz,1H),8.53(dd,J=5.1,1.8Hz,1H),8.49(d,J=8.7Hz ,1H),8.09(td,J=7.9,1.9Hz,1H),7.92–7.84(m,2H),7.80–7.77(m,1H),7.77–7.75(m,1H), 7.59(ddd,J=8.4,6.9,1.3Hz,1H),7.49–7.43(m,1H),5.90(s,1H),4.06(q,J=6.0Hz,2H),3. 62(q,J=6.1Hz,2H),2.94(d,J=6.0Hz,2H),2.68(d,J=4.2Hz,2H),1.82(q,J=3.6,3.0Hz,4H). 13 CNMR(101MHz,DMSO-d6)δ162.67,156.86,155.53,152.06,150.99,147.92,146.83,140.37,138.21,133.26,125.81,12 5.62,122.92,119.46,116.05,115.49,112.05,87.94,58.93,48.21,28.38,24.27,21.86,20.73.HRMS(m / z):calcdforC 24 H 25 N6O2[M+H] + ,429.2304;found,429.2024.

[0105] Example 22: Preparation of compound 5-hydroxy-1-phenyl-N-(3-((1,2,3,4-tetrahydroacridin-9-yl)amino)propyl)-1H-pyrazole-3-carboxamide (ET2)

[0106]

[0107] Compound 3a (204 mg, 1 mmol) was dissolved in DMF (15 mL), and DIPEA (528 μL, 3 mmol) was added. HATU (760 mg, 2 mmol) was added under ice bath, and compound 8b (255 mg, 1 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. Insoluble matter was precipitated after adding water (20 mL), which was filtered, and the filter cake was washed with water and dried. The target compound ET2 was purified by silica gel column chromatography as a white powder (300 mg) with a yield of 68%. 1 HNMR(400MHz,DMSO-d6)δ8.39(d,J=8.6Hz,1H),8.33(t,J=6.1Hz,1H),7.88–7 .81(m,2H),7.77(td,J=8.6,1.2Hz,3H),7.56–7.52(m,1H),7.52–7.47(m,2H) ,7.40–7.31(m,1H),5.85(s,1H),3.92(q,J=6.6Hz,2H),3.35–3.32(m,2H),2. 94(brs,2H),2.69–2.61(m,2H),1.98(p,J=6.5Hz,2H),1.80(q,J=3.4Hz,4H). 13 CNMR(101MHz,DMSO-d6))δ162.31,156.21,153.99,150.77,146.17,138.67,138.26,133.21,129.41(×2),127.15,125.57,1 25.51,122.43(×2),119.51,115.83,111.62,87.71,55.38,45.35,36.26,30.55,28.35,21.83,20.70.HRMS(m / z):calcdforC 26 H 28 N5O2[M+H] + ,442.2238;found,442.2262.

[0108] Example 23: Preparation of compound 5-hydroxy-1-phenyl-N-((1,2,3,4-tetrahydroacridin-9-yl)amino)ethyl-1H-pyrazole-3-carboxamide (ET3)

[0109]

[0110] Compound 3a (612 mg, 3 mmol) was dissolved in DMF (15 mL), and DIPEA (0.7 mL, 4 mmol) was added, followed by HATU (1425 mg, 3.75 mmol), and compound 8a (724 mg, 3 mmol). The mixture was reacted at room temperature for 18 h under N2 protection. The reaction solution was concentrated, 10 mL of water was added, and the mixture was extracted with ethyl acetate. The organic phase was concentrated and purified by silica gel column chromatography (DCM: MeOH = 30: 1, v / v) to obtain compound ET3, which was a brown solid of 800 mg. The yield was 62%. Melting point: 343°C–345°C. 1 HNMR(400MHz,DMSO-d6)δ8.53(d,J=6.0Hz,1H),8.50(d,J=8.8Hz,1H),7.92–7.88(m,1H), 7.87(d,J=7.5Hz,1H),7.79–7.77(m,1H),7.73(dd,J=7.9,1.5Hz,2H),7.59(ddd,J=8.5,6 .9,1.3Hz,1H),7.50(t,J=7.9Hz,2H),7.36(t,J=7.5Hz,1H),5.86(s,1H),4.06(q,J=6.0H z,2H),3.61(q,J=6.2Hz,2H),2.94(d,J=6.3Hz,2H),2.73–2.65(m,2H),1.86–1.77(m,4H). 13 CNMR(101MHz,DMSO-d6)δ163.04,156.84,154.03,150.96,145.79,138.60,138.22,133.27,129.42(×2),127.24,125.60 ,124.35,122.46(×2),119.45,116.04,112.02,87.75,67.71,48.38,28.39,21.87,20.74,20.47.HRMS(m / z):calcdforC 25 H 26 N5O2[M+H] + ,428.2081;found,428.2109.

[0111] Example 24: Preparation of compound 5-hydroxy-1-(pyridin-2-yl)-N-((3-(1,2,3,4-tetrahydroacridin-9-yl)amino)propyl)-1H-pyrazole-3-carboxamide (ET4)

[0112]

[0113] Compound 3b (205 mg, 1 mmol) was dissolved in DMF (15 mL), and DIPEA (528 μL, 3 mmol) was added. HATU (1140 mg, 3 mmol) was added under ice bath, and compound 8b (255 mg, 1 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. Insoluble matter was precipitated after adding water (10 mL), which was filtered, and the filter cake was washed with water and dried. The target compound ET4 was purified by silica gel column chromatography as a white powder (240 mg) with a yield of 54%. Melting point: 212°C–215°C. 1 HNMR(400MHz, DMSO-d6)δ13.26(s,1H),8.53(dd,J=5.1,1.8Hz,1H),8.42(t,J=6.1Hz,1H),8.39(d, J=8.7Hz,1H),8.09(td,J=7.9,1.9Hz,1H),7.86–7.80(m,2H),7.77(dd,J=8.5,1.3Hz,1H),7.54(ddd ,J=8.4,6.9,1.3Hz,1H),7.46(dd,J=7.5,5.1Hz,1H),5.90(s,1H),3.93(q,J=6.6Hz,2H),3.41–3.3 6(m,2H),2.94(d,J=4.8Hz,2H),2.67(d,J=5.5Hz,2H),2.00(p,J=6.6Hz,2H),1.80(p,J=2.7Hz,4H). 13 CNMR(101MHz,DMSO-d6)δ161.53,155.82,155.14,151.76,150.34,147.41,147.26,146.79,140.00,137.83,132.78,125.0 9,122.40,119.07,115.41,114.91,111.22,87.49,54.96,44.92,35.88,30.03,27.92,21.40,20.27.HRMS(m / z):calcdfor C 25 H 27 N6O2[M+H] + ,443.2190;found,443.2208.

[0114] Example 25: Preparation of compound N-(3-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)propyl)-5-hydroxy-1-phenyl-1H-pyrazole-3-carboxamide (ET5)

[0115]

[0116] Compound 3a (204 mg, 1 mmol) was dissolved in DMF (20 mL), and DIPEA (528 μL, 3 mmol) was added. HATU (950 mg, 2.5 mmol) was added under ice bath, and compound 8d (289 mg, 1 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. Insoluble matter was precipitated after adding water (25 mL), which was filtered, and the filter cake was washed with water and dried. The target compound ET5 was purified by silica gel column chromatography as a white solid (285 mg) with a yield of 60%. Melting point: 225°C–228°C. 1 HNMR(400MHz, DMSO-d6)δ8.32(d,J=9.2Hz,1H),8.29(s,1H),7.84–7.76(m,2H),7.73(d,J=2.2Hz,1H),7.51–7.44(m,3H),7.40(s,1H),7.3 3(d,J=7.1Hz,1H),5.79(s,1H),3.79(d,J=6.5Hz,2H),3.31(d,J=6.3Hz,2H),2.89(s,2H),2.63(s,2H),1.92(d,J=6.5Hz,2H),1.77(s,4H). 13 CNMR(101MHz,DMSO-d6)δ162.41,154.83,146.15,141.28,138.91,136.44,129.34(×2),128.87,127.53,126.87,125.21,12 4.19,122.20(×2),120.49,115.52,113.19,87.46,45.44,36.24,30.48,29.77,24.48,22.03,21.13.HRMS(m / z):calcdforC 26 H 27 ClN5O2[M+H] + ,476.1848;found,476.1840.

[0117] Example 26: Preparation of Compound N-(3-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)propyl)-5-hydroxy-1-pyridin-2-yl-1H-pyrazole-3-carboxamide (ET6)

[0118]

[0119] Compound 3b (205 mg, 1 mmol) was dissolved in DMF (15 mL), and DIPEA (264 μL, 1.5 mmol) was added. HATU (448 mg, 1.17 mmol) was added under ice bath, and compound 8d (289 mg, 1 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. Insoluble matter was precipitated after adding water (15 mL), which was filtered, and the filter cake was washed with water and dried. The target compound ET6 was purified by silica gel column chromatography as a white powder (380 mg) with a yield of 79%. Melting point: 215°C–216°C. 1 HNMR(400MHz,DMSO-d6)δ8.53(dd,J=5.0,1.7Hz,1H),8.41(t,J=6.0Hz,1H),8.37(d,J=9.3Hz,1H),8 .10(td,J=7.9,1.9Hz,1H),7.96(d,J=6.0Hz,1H),7.80(d,J=8.4Hz,1H),7.71(d,J=2.2Hz,1H),7.52 (dd,J=9.2,2.2Hz,1H),7.47(ddd,J=7.5,5.0,1.0Hz,1H),5.88(s,1H),3.92(q,J=6.5Hz,2H),3.36( q,J=6.3Hz,2H),2.91(t,J=5.5Hz,2H),2.62(brs,2H),2.00(p,J=6.5Hz,2H),1.79(t,J=3.6Hz,4H). 13 CNMR(101MHz,DMSO-d6)δ161.85,156.14,155.58,152.20,151.12,147.78,147.14,140.43,139.03,137.61,128.11,125.6 1,122.78,118.24,115.18,114.38,112.18,87.87,45.46,36.25,30.12,28.37,24.13,21.69,20.62.HRMS(m / z):calcdfor C 25 H 26 ClN6O2[M+H] + ,477.1800;found,477.1804.

[0120] Example 27: Preparation of Compound N-(2-((2,3-dihydro-1H-cyclopentadienyl[b]quinolin-9-yl)amino)ethyl)-5-hydroxy-1-(pyridin-2-yl)-1H-pyrazole-3-carboxamide (ET7)

[0121]

[0122] Compound 3b (205 mg, 1 mmol) was dissolved in DMF (15 mL), and DIPEA (352 μL, 2 mmol) was added. HATU (448 mg, 1.17 mmol) was added under ice bath, and compound 8g (227 mg, 1 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. Insoluble matter was precipitated after adding water (10 mL), which was filtered, and the filter cake was washed with water and dried. The target compound ET7 was purified by silica gel column chromatography as a white powder (300 mg) with a yield of 72%. Melting point: 160°C–163°C. 1 HNMR(600MHz,DMSO-d6)δ8.69(dt,J=12.7,5.8Hz,2H),8.52(dd,J=12.0,5.1Hz,1H),8.43(d ,J=9.0Hz,1H),8.09(t,J=7.5Hz,1H),7.87(q,J=6.8,5.8Hz,1H),7.78(dd,J=16.4,8.2Hz,2 H),7.66(t,J=7.8Hz,1H),7.45(dq,J=13.3,7.1,6.0Hz,1H),5.92(s,1H),3.91(q,J=6.5Hz, 2H),3.57(q,J=6.5Hz,2H),3.47–3.40(m,2H),3.11(t,J=7.8Hz,2H),2.16(p,J=7.6Hz,2H). 13 CNMR(151MHz,DMSO-d6)δ162.43,161.91,161.37,155.57,153.21,152.11,147.89,147.04,140.38,137.71,132.98 ,126.43,123.59,122.88,119.99,117.30,115.43,88.01,44.70,39.01,31.64,31.34,22.73.HRMS(m / z):calcdforC 23 H 21 N6O2[MH] + ,413.1731;found,413.1733.

[0123] Example 28: Preparation of Compound N-(3-((2,3-dihydro-1H-cyclopentadienyl[b]quinolin-9-yl)amino)propyl)-5-hydroxy-1-(pyridin-2-yl)-1H-pyrazole-3-carboxamide (ET8)

[0124]

[0125] Compound 3b (205 mg, 1 mmol) was dissolved in DMF (15 mL), and DIPEA (528 μL, 3 mmol) was added. HATU (622 mg, 1.6 mmol) was added under ice bath, and compound 8h (241 mg, 1 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. Insoluble matter was precipitated after adding water (20 mL), which was filtered, and the filter cake was washed with water and dried. The target compound ET8 was purified by silica gel column chromatography as a white powder (270 mg) with a yield of 63%. Melting point: 192°C–194°C. 1 HNMR(600MHz,DMSO-d6)δ8.66(t,J=6.5Hz,1H),8.56–8.50(m,1H),8.47(t,J=6.2Hz,1H),8.41(d ,J=8.6Hz,1H),8.10(td,J=7.9,1.9Hz,1H),7.84(q,J=7.7Hz,2H),7.75(d,J=8.4Hz,1H),7.62(t ,J=7.8Hz,1H),7.46(dd,J=7.4,5.0Hz,1H),5.93(s,1H),3.78(q,J=6.9Hz,2H),3.39(d,J=6.3Hz ,2H),3.29(t,J=7.4Hz,2H),3.11(t,J=7.9Hz,2H),2.13(p,J=7.6Hz,2H),1.93(p,J=6.8Hz,2H). 13 CNMR(151MHz,DMSO-d6)δ162.76,161.94,161.31,155.62,152.91,152.27,147.77,147.27,140.45,137.69,132.95,12 6.41,123.38,122.77,120.00,117.21,115.22,87.93,42.37,36.19,31.63,31.44,31.39,22.68.HRMS(m / z):calcdforC 24 H 23 N6O2[MH] + ,427.1888;found,427.1888.

[0126] Example 29: Preparation of Compound N-(2-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)ethyl)-5-hydroxy-1-(pyridin-2-yl)-1H-pyrazole-3-carboxamide (ET9)

[0127]

[0128] Compound 3b (205 mg, 1 mmol) was dissolved in DMF (15 mL), and DIPEA (528 μL, 3 mmol) was added. HATU (829 mg, 2.1 mmol) was added under ice bath, and compound 8c (275 mg, 1 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. Insoluble matter was precipitated after adding water (25 mL), which was filtered and dried. The target compound ET9 was purified by silica gel column chromatography, and the product was 200 mg of white powder with a yield of 43%. Melting point: 199°C–203°C. 1 HNMR(600MHz,DMSO-d6)δ8.53(d,J=4.5Hz,2H),8.50(d,J=9.2Hz,1H),8.09(t,J=7.8 Hz,1H),8.00(brs,1H),7.74(d,J=8.3Hz,1H),7.72(d,J=2.0Hz,1H),7.60(dd,J=9.1 ,2.1Hz,1H),7.47(dd,J=7.4,5.0Hz,1H),5.89(s,1H),4.05(q,J=6.0Hz,2H),3.61(q ,J=6.1Hz,3H),2.90(q,J=4.6,3.1Hz,2H),2.65(d,J=6.1Hz,2H),1.82–1.79(m,4H). 13 CNMR(151MHz,DMSO-d6)δ162.64,156.90,155.57,152.08,151.52,147.89,146.75,140.34,139.00,137.65,128.34,12 5.77,122.91,118.30,115.40,114.73,112.71,87.89,54.07,48.30,42.31,28.45,21.73,20.65.HRMS(m / z):calcdforC 24 H 22 ClN6O2[MH] + ,461.1498;found,461.1544.

[0129] Example 30: Preparation of Compound N-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)butyl)-5-hydroxy-1-(pyridin-2-yl)-1H-pyrazole-3-carboxamide (ET10)

[0130]

[0131] Compound 3b (410 mg, 2 mmol) was dissolved in DMF (5 mL), and DIPEA (1045 μL, 6 mmol) was added under ice bath, HATU (912 mg, 2.4 mmol) was added under ice bath, and compound 8e (667 mg, 2.2 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. Water (30 mL) was added to precipitate a light yellow solid, which was filtered, and the filter cake was washed with methanol, dried, and purified by silica gel column chromatography to obtain compound ET10 as a light yellow solid (200 mg) with a yield of 20%. Melting point: 194°C–196°C. 1 HNMR(600MHz,DMSO-d6)δ12.91(s,1H),8.51(d,J=3.5Hz,1H),8.38(d,J=9.2Hz,1H),8.22(t,J=6.0 Hz,1H),8.08(td,J=7.9,1.8Hz,1H),7.82(s,1H),7.79(d,J=8.2Hz,1H),7.71(d,J=2.1Hz,1H),7.5 5(dd,J=9.2,2.1Hz,1H),7.47–7.42(m,1H),5.87(s,1H),3.88(d,J=6.7Hz,2H),3.25(d,J=6.4Hz,2 H),2.92–2.91(m,2H),2.61(s,2H),1.82–1.80(m,4H),1.77(p,J=7.2Hz,2H),1.61(p,J=6.9Hz,2H). 13 CNMR(151MHz,DMSO-d6)δ161.08,155.56,155.19,151.91,151.24,147.21,146.92,140.01,138.67,137.15,127.72,125.23,1 22.21,117.92,114.53,114.10,111.85,87.42,47.05,37.97,28.02,27.00,26.04,23.70,21.27,20.20.HRMS(m / z):calcdforC 26 H 26 ClN6O2[MH] + ,489.1811;found,489.1813.

[0132] Example 31: Preparation of compound N-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)propyl)-1-(5-fluoropyridin-2-yl)-5-hydroxy-1H-pyrazole-3-carboxamide (ET11)

[0133]

[0134] Compound 3c (223 mg, 1 mmol) was dissolved in DMF (5 mL), and DIPEA (525 μL, 3 mmol) was added under ice bath, HATU (456 mg, 1.2 mmol) was added under ice bath, and compound 8d (300 mg, 1.03 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. After adding water (30 mL), a muddy brown solid was precipitated, and the filtrate was extracted with DCM, concentrated and dried, and the filter cake and the organic phase were combined to make sand. Compound ET11 was obtained by silica gel column chromatography with DCM:MeOH (20:1, V / V) as a white solid (50 mg) with a yield of 10%. Melting point: 224°C–227°C. 1 HNMR(600MHz,DMSO-d6)δ8.56(d,J=3.1Hz,1H),8.38–8.36(m,2H),8.01(td,J=8. 6,3.1Hz,1H),7.93(s,1H),7.79(dd,J=9.0,3.9Hz,1H),7.74(d,J=2.2Hz,1H),7. 53(dd,J=9.2,2.2Hz,1H),5.84(s,1H),3.90(q,J=6.6Hz,2H),3.39–3.28(m,2H), 2.91(t,J=5.6Hz,2H),2.62(s,2H),1.97(p,J=6.6Hz,2H),1.79(p,J=2.9Hz,4H). 13 CNMR(151MHz,DMSO-d6)δ161.13,157.38(d,J=252.7Hz),155.29,153.88,1 50.31,146.74,138.20,136.77,135.44(d,J=26.3Hz),127.25,126.06(d,J= 19.8Hz), 124.80, 117.96 (d, J = 4.5Hz), 117.94, 117.42, 113.56, 111.33, 86. 87,44.55,35.33,29.29,27.51,23.25,20.82,19.75.HRMS(m / z):calcdforC 25 H 23 ClFN6O2[MH] + ,493.1561;found,493.1561.

[0135] Example 32: Preparation of compound N-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)butyl)-1-(5-fluoropyridin-2-yl)-5-hydroxy-1H-pyrazole-3-carboxamide (ET12)

[0136]

[0137] Compound 3c (223 mg, 1 mmol) was dissolved in DMF (5 mL), and DIPEA (525 μL, 3 mmol) was added under ice bath, HATU (456 mg, 1.2 mmol) was added under ice bath, and compound 8e (300 mg, 0.99 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. Water (30 mL) was added to precipitate a gray solid, which was filtered and extracted with DCM. The filtrate was concentrated and dried. The filter cake and the organic phase were combined to make sand, and the compound ET12 was obtained by silica gel column chromatography with DCM:MeOH (20:1, V / V) as a white solid (90 mg) with a yield of 17%. Melting point: 196°C–198°C. 1 HNMR (600MHz, DMSO-d6) δ8.54(d,J=3.0Hz,1H),8.38(d,J=9.3Hz,1H),8.20(t,J=6.0Hz,1H),7. 99(td,J=8.6,3.0Hz,1H),7.81(s,1H),7.79(dd,J=9.1,4.0Hz,1H),7.72(d,J=2.2Hz,1H),7.55( dd,J=9.2,2.2Hz,1H),5.84(s,1H),3.87(q,J=6.7Hz,2H),3.23(q,J=6.5Hz,2H),2.92(q,J=5.3 ,4.0Hz,2H),2.62–2.59(m,2H),1.82–1.80(m,4H),1.75(p,J=7.3Hz,2H),1.59(p,J=6.9Hz,2H). 13 CNMR(151MHz,DMSO-d6)δ161.49,157.95(d,J=252.6Hz),155.84,154.59,151.36,147.49,146.87,138.84,137.44,135.93(d,J=26.2Hz),129.07, 127.97,126.74(d,J=19.8Hz),125.48,118.08,114.27,112.06,87.55,5 3.86,47.29,38.17,28.19,27.22,26.25,21.48,20.40.HRMS(m / z):calcd forC 26 H 25 ClFN6O2[MH] + ,507.1717;found,507.1722.

[0138] Example 33: Preparation of compound N-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)ethyl)-1-(5-fluoropyridin-2-yl)-5-hydroxy-1H-pyrazole-3-carboxamide (ET13)

[0139]

[0140] Compound 3c (446 mg, 2 mmol) was dissolved in DMF (5 mL), and DIPEA (1045 μL, 6 mmol) was added under ice bath, HATU (912 mg, 2.4 mmol) was added under ice bath, and compound 8c (661 mg, 2.4 mmol) was added. The reaction was carried out at room temperature for 24 h under N2 protection. The reaction solution was concentrated at 85 °C, and 10 mL of methanol was added to the reaction solution. A light yellow solid was precipitated, which was filtered, and the filter cake was sanded. Compound ET13 was obtained by silica gel column chromatography DCM: MeOH (50:1, V / V), a white flaky solid (250 mg), with a yield of 27%. Melting point: 190 °C–192 °C. 1 HNMR (600MHz, DMSO-d6) δ12.80 (s, 1H), 8.56 (d, J = 3.3Hz, 1H), 8.55 (t, J = 6. 4Hz,1H),8.49(d,J=9.2Hz,1H),8.03–7.98(m,2H),7.80–7.73(m,2H),7.58 (dd,J=9.1,2.2Hz,1H),5.86(s,1H),4.03(q,J=6.0Hz,2H),3.60(q,J=6.1H z, 2H), 2.91 (t, J = 5.8Hz, 2H), 2.65 (t, J = 5.7Hz, 2H), 1.81 (q, J = 6.8Hz, 4H). 13 CNMR(151MHz,DMSO-d6)δ162.77,158.30(d,J=252.9Hz),156.83,154.74, 151.44,147.44,146.51,138.95,137.68,136.38(d,J=26.2Hz),128.35,1 26.86(d,J=20.3Hz),125.75,119.00(d,J=4.2Hz),118.24,114.67,112.6 2,87.73,54.05,48.32,42.31,28.41,21.72,20.63.HRMS(m / z):calcdforC 24 H 23 ClFN6O2[M+H] + ,481.1550;found,481.1570.

[0141] Example 34: Preparation of Compound N-((6-chloro-2,3-dihydro-1H-cyclopenta[b]quinolin-9-yl)amino)propyl)-1-(5-fluoropyridin-2-yl)-5-hydroxy-1H-pyrazole-3-carboxamide (ET14)

[0142]

[0143] Compound 3c (223 mg, 1 mmol) was dissolved in DMF (5 mL), and DIPEA (525 μL, 3 mmol) was added under ice bath, HATU (456 mg, 1.2 mmol) was added under ice bath, and compound 8f (303 mg, 1.1 mmol) was added. The reaction was allowed to react at room temperature for 24 h under N2 protection. Water was added to quench the reaction, and a black solid precipitated. The filter cake was dissolved in 10 mL of DCM, and petroleum ether was added dropwise. A white solid precipitated, and the mixture was filtered to obtain compound ET14, a white powder solid (50 mg), with a yield of 10%. Melting point: 168°C–170°C. 1 HNMR(600MHz,DMSO-d6)δ8.74(t,J=6.2Hz,1H),8.56(d,J=2.9Hz,1H),8.43(d,J=9.2Hz,1H),8 .39(t,J=6.1Hz,1H),8.01(td,J=8.5,3.0Hz,1H),7.79(dd,J=9.1,4.0Hz,1H),7.72(d,J=2.0H z,1H),7.68(dd,J=9.0,2.1Hz,1H),5.86(s,1H),3.76(q,J=6.8Hz,2H),3.36(q,J=6.5,5.6Hz, 2H), 3.27 (t, J = 7.3Hz, 2H), 3.10 (t, J = 7.9Hz, 2H), 2.12 (p, J = 7.7Hz, 2H), 1.91 (p, J = 6.8Hz, 2H). 13 CNMR(151MHz,DMSO-d6)δ161.57,161.50,157.78(d,J=252.8Hz),154.33, 152.36,147.19,146.55,138.12,137.02,135.80(d,J=26.2Hz),126.50(d ,J=20.5Hz),126.20,125.36,118.47,118.29(d,J=4.5Hz),115.61,112.4 5,87.30,42.12,35.74,31.28,30.78,30.71,22.15.HRMS(m / z):calcdforC 24 H 21 ClFN6O2[MH] +,479.1404;found,479.1407.

[0144] Example 35: Preparation of Compound N-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)-1-(5-chloropyridin-2-yl)-5-hydroxy-1H-pyrazole-3-carboxamide (ET15)

[0145]

[0146] Compound 3d (288 mg, 1.2 mmol) was dissolved in DMF (5 mL), and DIPEA (627 μL, 3.6 mmol) was added under ice bath, HATU (912 mg, 2.4 mmol) was added under ice bath, and compound 8c (331 mg, 1.2 mmol) was added. Under N2 protection, the reaction was allowed to react at room temperature for 24 h. Water was added to quench the reaction, and a gray solid was precipitated. The filter cake was filtered, and the filter cake was washed three times with methanol (10 mL). The filter cake was sonicated and filtered to obtain compound ET15 as a white solid (256 mg) with a yield of 43%. Melting point: 154°C–164°C. 1 HNMR (600MHz, DMSO-d6) δ8.52(d,J=2.6Hz,1H),8.37(t,J=6.0Hz,1H),8.30(d,J=9. 2Hz,1H),8.06(dd,J=8.8,2.6Hz,1H),8.00(d,J=8.8Hz,1H),7.70(d,J=2.3Hz,1H), 7.40(dd,J=9.1,2.2Hz,1H),6.72(s,1H),5.59(s,1H),3.78(q,J=5.9Hz,2H),3.52( q,J=6.2Hz,2H),2.86(d,J=5.8Hz,2H),2.66(d,J=5.4Hz,2H),1.76(t,J=3.7Hz,4H). 13 CNMR(151MHz,DMSO-d6)δ163.04,157.87,153.58,150.17,147.23,146.81,143.89,138.84,135.03,128.55,127.08,12 4.69,123.06,120.35,117.62,116.93,114.64,86.87,48.51,40.51,31.42,24.82,22.43,21.74.HRMS(m / z):calcdforC 24 H 23 Cl2N6O2[M+H] + ,497.1254;found,497.1276.

[0147] Example 36: Preparation of Compound N-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)propyl)-5-hydroxy-1-(5-(trifluoromethyl)pyridin-2-yl)-1H-pyrazole-3-carboxamide (ET16)

[0148]

[0149] Compound 3f (409 mg, 1.5 mmol) was dissolved in DMF (5 mL), and DIPEA (785 μL, 4.5 mmol) was added under ice bath, HATU (684 mg, 1.8 mmol) was added under ice bath, and compound 8d (433 mg, 1.5 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. The DMF was removed by concentration at 85 °C, and water (5 mL) and methanol (10 mL) were added. A gray solid was precipitated, which was filtered, and the filter cake was sanded. Compound ET16 was obtained by silica gel column chromatography with DCM:MeOH (30:1, V / V) as a light yellow solid (100 mg) with a yield of 12%. Melting point: 195 °C–198 °C. 1 HNMR(600MHz,DMSO-d6)δ8.91(s,1H),8.56–8.52(m,1H),8.42(d,J=8.7Hz,1H),8.38(d,J=8.9Hz,1H),8.13(s,1H),8.01–7.95(m,2H),7.48(d ,J=9.0Hz,1H),5.85(s,1H),3.93–3.85(m,2H),2.97–2.93(m,2H),2.65 –2.59(m,2H),2.04–1.94(m,2H),1.79–1.74(m,4H),1.23–1.19(m,2H). 13 CNMR(151MHz,DMSO-d6)δ161.98,155.72(×2),154.26,151.45,148.16,145.75,139.32,137.17,137.08,127.94,125.47,124.06(q,J =271.3Hz),122.97,118.50,115.69,114.51,112.12,87.93,45.26,36.26,30.20,28.46,24.29,21.77,20.62.HRMS(m / z):calcdforC 26 H 25 ClF3N6O2[M+H] + ,545.1674;found,545.1696.

[0150] Example 37: Preparation of Compound N-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)propyl)-5-hydroxy-1-(pyrimidin-2-yl)-1H-pyrazole-3-carboxamide (ET17)

[0151]

[0152] Compound 3e (412 mg, 2 mmol) was dissolved in anhydrous dichloromethane (10 mL), triethylamine (832 μL, 6 mmol) was added under ice bath, EDCI (573 mg, 3 mmol) was added under ice bath, HOBT (405 mg, 3 mmol) was added, compound 8d (578 mg, 2 mmol) was added, and the reaction was carried out at room temperature for 24 h under N2 protection. The solvent was dried, 10 mL of water and methanol were added, and a muddy solid was precipitated. The filtrate was filtered, and sand was made. Compound ET17 was obtained by silica gel column chromatography DCM: MeOH (20:1, V / V), a light yellow solid powder (50 mg), and a yield of 5%. Melting point: 101°C–103°C. 1 HNMR(600MHz,DMSO-d6)δ8.96–8.84(m,2H),8.30(t,J=6.1Hz,1H),8.23(d,J =9.2Hz,1H),7.79(d,J=30.8Hz,1H),7.52(dd,J=29.8,6.7Hz,1H),7.39(d,J= 9.1Hz,1H),7.27(dt,J=36.9,7.9Hz,1H),5.72(s,1H),3.66–3.58(m,2H),3.2 9(q,J=6.7Hz,2H),2.88(s,2H),2.64(s,2H),1.88–1.80(m,2H),1.76(s,4H). 13 CNMR(151MHz,DMSO-d6)δ161.97,158.86(×2),158.56,156.41,147.60,142.92,134.17,127.66,126.15,125.07,125 .04,124.14,123.40,118.60,110.34,86.59,48.61,44.82,35.76,30.32,24.53,22.06,21.42.HRMS(m / z):calcdfor C 24 H 25 ClN7O2[M+H] + ,478.1753;found,478.1774.

[0153] Example 38: Preparation of Compound N-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)ethyl)-5-hydroxy-1-(5-(trifluoromethyl)pyridin-2-yl)-1H-pyrazole-3-carboxamide (ET18)

[0154]

[0155] Compound 3f (409 mg, 1.5 mmol) was dissolved in DMF (5 mL), and DIPEA (785 μL, 4.5 mmol) was added under ice bath, HATU (684 mg, 1.8 mmol) was added under ice bath, and compound 8c (413 mg, 1.5 mmol) was added. The mixture was reacted at room temperature for 24 h under N2 protection. DMF was removed by concentration at 85 °C, and water (5 mL) and methanol (10 mL) were added. A muddy solid was precipitated, which was filtered, and the filter cake was sanded. Compound ET18 was obtained by silica gel column chromatography with DCM:MeOH (50:1, V / V) as a white solid (200 mg) with a yield of 25%. Melting point: 156 °C–163 °C. 1 HNMR(600MHz,DMSO-d6)δ8.94(d,J=2.4Hz,1H),8.62(t,J=6.1Hz,1H),8.53– 8.43(m,2H),8.01(t,J=5.6Hz,1H),7.98(d,J=8.7Hz,1H),7.70(d,J=2.1Hz,1 H),7.59(dd,J=9.2,2.1Hz,1H),5.91(s,1H),4.05(q,J=6.0Hz,2H),3.62(q,J =6.1Hz,2H),2.93–2.87(m,2H),2.65(d,J=6.2Hz,2H),1.81(q,J=6.8Hz,4H). 13 CNMR(151MHz,DMSO-d6)δ162.44,156.89,155.82,154.16,151.47,151.44,14 7.67,145.77(q,J=3.1Hz),138.93,137.65,137.36(q,J=3.6Hz),128.30,125 .75,123.97(q,J=272.2Hz),123.61(q,J=32.1,31.6Hz),118.23,115.88,114 .68,112.68,88.37,48.23,28.41,24.17,21.71,20.63.HRMS(m / z):calcdforC 25 H 23 ClF3N6O2[M+H] +,531.1518;found,531.1537.

[0156] Example 39: Preparation of Compound N-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)ethyl)-5-hydroxy-1-(pyrimidin-2-yl)-1H-pyrazole-3-carboxamide (ET19)

[0157]

[0158] Compound 3e (370 mg, 1.8 mmol) was dissolved in anhydrous dichloromethane (10 mL), triethylamine (625 μL, 4.5 mmol) was added under ice bath, EDCI (429 mg, 2.25 mmol) was added under ice bath, HOBT (303 mg, 2.25 mmol) was added, compound 8c (413 mg, 1.5 mmol) was added, and the reaction was carried out at room temperature for 24 h under N2 protection. The solvent was dried, 10 mL of water was added, and a red solid was precipitated, which was filtered, and the filter cake was sanded. Compound ET19 was obtained by silica gel column chromatography DCM: MeOH (10:1, V / V), pink solid powder (120 mg), yield 14%. Melting point: 158°C–171°C. 1 HNMR(600MHz,DMSO-d6)δ8.89(d,J=4.9Hz,2H),8.41(t,J=6.1Hz,1H),8.25(d, J=9.1Hz,1H),7.73(d,J=2.2Hz,1H),7.52(t,J=4.9Hz,1H),7.37(dd,J=9.0,2. 3Hz,1H),6.31–6.20(m,1H),5.75(s,1H),3.70(q,J=6.1Hz,2H),3.50(q,J=6.3 Hz,2H),2.89–2.85(d,J=6.3Hz,2H),2.69(q,J=5.6Hz,2H),1.80–1.75(m,4H). 13 CNMR(151MHz,DMSO-d6)δ163.06,162.27,158.89,157.23,156.32×2,151.58,147.38,145.74,133.34 ,126.02,124.87,123.76,119.72,117.53,115.09,86.81,47.99,32.39,24.66,22.30,21.81,20.77.

[0159] Example 40: Preparation of Compound N-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)-1-(3-chloropyridin-2-yl)-5-hydroxy-1H-pyrazole-3-carboxamide (ET20)

[0160]

[0161] Compound 3g (718 mg, 3 mmol) was dissolved in anhydrous dichloromethane (10 mL), triethylamine (832 μL, 6 mmol) was added under ice bath, EDCI (573 mg, 3 mmol) was added under ice bath, HOBT (405 mg, 3 mmol) was added, compound 8c (413 mg, 1.5 mmol) was added, and the reaction was carried out at room temperature for 24 h under N2 protection. The solvent was dried, 10 mL of water was added, and a brown oil was precipitated, extracted with DCM, and the organic phase was combined to make sand. Compound ET20 was obtained by silica gel column chromatography with DCM: MeOH (25:1, V / V), as a gray solid (130 mg), with a yield of 13%. Melting point: 178°C–180°C. 1 HNMR(600MHz,DMSO-d6)δ8.60(dd,J=4.7,1.6Hz,1H),8.56(td,J=6.0,2.1Hz,1H),8.48 (d,J=9.2Hz,1H),8.24(dd,J=8.1,1.5Hz,1H),7.99–7.95(m,1H),7.93–7.85(m,1H),7. 66(dd,J=8.1,4.6Hz,1H),7.54(dt,J=9.4,2.1Hz,1H),5.85(s,1H),3.99(q,J=6.0Hz,2 H),3.57(q,J=6.1Hz,2H),2.99–2.93(m,2H),2.65(d,J=5.4Hz,2H),1.83–1.77(m,4H). 13 C NMR(151MHz,DMSO-d6)δ162.49,155.78,154.64,151.46,147.72,146.69,146.49,139.66,139.04,136.83,128.84,127 .74,126.78,125.13,118.28,114.43,112.15,85.79,47.81,40.06,28.19,23.76,21.36,20.26.HRMS(m / z):calcdforC 24 H 23 Cl2N6O2[M+H] + ,497.1254;found,497.1278.

[0162] Example 41: Preparation of compound N-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)propyl)-1-(3-chloropyridin-2-yl)-5-hydroxy-1H-pyrazole-3-carboxamide (ET21)

[0163]

[0164] Compound 3g (480 mg, 2 mmol) was dissolved in DMF (7 mL), HOBT (405 mg, 3 mmol) was added under ice bath, pybop (1.56 g, 3 mmol) was added, and compound 8d (578 mg, 2 mmol) was added. The reaction was carried out at room temperature for 24 h under N2 protection. The solvent was dried and 10 mL of water was added. A black oil was precipitated. The mixture was extracted with ethyl acetate, and the organic phases were combined for sand making. The compound ET21 was obtained as a brown solid (100 mg) by silica gel column chromatography with DCM:MeOH (50:1, V / V). The yield was 10%. Melting point: 175°C–177°C. 1 HNMR(600MHz,DMSO-d6)δ11.90(s,1H),8.60(d,J=4.6Hz,1H),8.39(t,J=6.1Hz,1H),8 .37(d,J=9.0Hz,1H),8.24(d,J=8.0Hz,1H),7.78(d,J=2.0Hz,1H),7.67(dd,J=8.1,4. 7Hz,1H),7.53(d,J=9.2Hz,1H),5.81(s,1H),3.88(q,J=6.7Hz,2H),3.30(q,J=6.2Hz, 2H), 2.93 (t, J = 6.0Hz, 2H), 2.61 (s, 2H), 1.94 (p, J = 6.6Hz, 2H), 1.80 (t, J = 4.0Hz, 4H). 13 CNMR(151MHz,DMSO-d6)δ162.58,161.99,155.89,154.74,151.41,148.00,147.21,146.96,139.96,137.49,129.13,127.0 7,125.53,121.01,118.19,114.22,111.99,86.04,45.12,42.11,35.87,30.13,28.22,21.53,20.46.HRMS(m / z):calcdfor C 25 H 25 Cl2N6O2[M+H] + ,511.1411;found,511.1433.

[0165] Example 42: Acetylcholinesterase (AChE) inhibition ability experiment

[0166] Preparation of acetylcholinesterase inhibition ability test solution: Accurately weigh 680 mg of KH2PO4 and dissolve it in 50 mL of distilled water, adjust the pH to 8±0.1 with KOH, and store the resulting buffer solution in a 4°C refrigerator for later use. Accurately weigh 15 mg of NaHCO3 and 39.6 mg of DTNB, and dissolve them in 10 mL of distilled water to obtain a 10 mM DTNB solution, and store the solution in a -20°C refrigerator for later use. Accurately weigh 217 mg of ATC and dissolve it in 10 mL of distilled water to obtain a 75 mM ATC solution, and store the solution in a -20°C refrigerator for later use. Dissolve 500 units of AChE in 50 mL of distilled water to obtain an AChE solution with a concentration of 10 units / mL, divide it into 2 mL centrifuge tubes, and store it in a -20°C refrigerator for later use. Prepare the compound to be tested with DMSO to 20 mM, and prepare a concentration of 10 with distilled water. -4 The mother solution was then diluted with distilled water to prepare a concentration of 10 -5 , 10 -6 , 10 -7 , 10 -8 , 10 -9 M solution, freshly prepared and used. Place all the above solutions at room temperature one hour in advance, and dilute the AChE solution with a concentration of 10 units / mL with distilled water to an AChE solution with a concentration of 2.5 units / mL. The specific addition is shown in Table 1. Donepezil is the positive control compound. The absorbance (OD) is then measured at a wavelength of 412nm. 10μL of distilled water without the test compound is used as the control group. 20μL of distilled water without the test compound and enzyme is used as the blank group, and the above tests are performed in parallel three times. The ultraviolet absorption value measured in the control group is used as the baseline value of 100%, and the inhibition rate of the test compound at each concentration is calculated. The calculation results are used to calculate the IC of the test compound using GraphPad Prism TM (GraphPad Software, San Diego, CA, USA) software in non-linear regression analysis mode (non-linear regression analysis model). 50 The experimental results (Table 2) show that more than half of the compounds have inhibitory activity against AChE, some of which show better AChE inhibitory activity than donepezil, and some compounds have AChEIC 50 Close to 1nM (donepezil AChEIC 50 Close to 40nM).

[0167] Table 1 AChE test sample addition table

[0168] Group Buffer Test compound AChE DTNB ATC Total volume Control group 40μL 10 μL distilled water 10μL 20μL 20μL 100μL Blank Group 40μL 10 μL distilled water 10 μL distilled water 20μL 20μL 100μL Compound Group 40μL 10μL 10μL 20μL 20μL 100μL

[0169] Table 2 Inhibitory effect of compounds on acetylcholinesterase

[0170]

[0171] Note: ND in the table means not tested.

[0172] Example 43: DPPH free radical scavenging experiment

[0173] Preparation of DPPH test solution: 1.35 mg of DPPH solid was dissolved in 34.3 mL of methanol to prepare a 100 μM DPPH test solution. Preparation of sample solution: The sample was dissolved in a suitable solvent to prepare a 30 mM solution. The specific addition is shown in Table 3. Before the test, the 30 mM solution of the test compound was diluted with methanol to 300 μM, and Edaravone (Eda) was used as a positive control compound. It was then diluted with methanol to 5 concentrations (10, 20, 40, 60, 80 μM) for testing. A value measurement (test group): In a 96-well plate, 40 μL of the methanol solution of the test compound was taken with a pipette, and then 160 μL of DPPH solution was added. All the above tests were performed twice in parallel. At room temperature for 10 hours, the A519 nm value was measured. A0 value measurement (blank group): 40 μL of methanol solution was taken with a pipette, and 160 μL of DPPH solution was added. The measured A value is A0. The experimental results (Table 4) show that most compounds have scavenging ability against DPPH at concentrations of 4 to 16 μM. Among them, most of the target compounds show better DPPH scavenging ability than edaravone. The scavenging ability of some compounds against DPPH at a concentration of 16 μM reaches more than 60% (the scavenging rate of edaravone at the same concentration is 35.8%).

[0174] Table 3 DPPH test sample addition table

[0175] Test compound solution Methanol DPPH test solution Total volume 30μL 10μL 160μL 200μL 25μL 15μL 160μL 200μL 20μL 20μL 160μL 200μL 10μL 30μL 160μL 200μL 5μL 35μL 160μL 200μL

[0176] Table 4 DPPH scavenging ability of the compounds (%)

[0177]

[0178]

[0179] Example 44: Evaluation of the protective effect of tert-butyl hydroperoxide on neuronal cell damage

[0180] The selection of neural cells includes but is not limited to SH-SY5Y and PC-12 cells. The experimental method is: take a bottle of cells in the logarithmic growth phase, add trypsin cell digestion solution to digest, so that the attached cells fall off, and prepare a concentration of 1×10 5100 μL of cell suspension (1×10 cells / mL) was added to each well of a 96-well cell culture plate. 4 cells), and the plate was placed in a 37°C, 5% CO2 incubator for 24 hours. Cell damaging agents, such as tert-butyl hydroperoxide (tBHP), were prepared with DMEM culture medium. The preferred tBHP concentration for this test was 550 μM. The compound to be tested was then prepared with DMEM culture medium containing the cell damaging agent, and the culture medium containing the compound to be tested was incubated with the corresponding cells for 36 hours. The culture medium was then discarded, the plate was washed once with PBS buffered saline, and 100 μL of the corresponding DMEM (containing damaging agent) culture medium containing different concentrations of the compound to be tested was added to each well; a blank control group (only high-glucose DMEM culture medium) was set up at the same time. The 96-well plate was placed in a 37°C, 5% CO2 incubator for 48 hours, 15 μL of MTT solution (5 mg / mL) was added to each well, and the culture was continued in the incubator for 4 hours. The culture medium was discarded, 150 μL of DMSO was added to each well to dissolve, and the crystals were completely dissolved by shaking on a shaker for 5 minutes. Finally, the OD value was read at a wavelength of 490 nm using a multifunctional microplate reader to calculate the cell growth inhibition rate. The results showed that the survival rate of cells under the action of 550 μM tBHP was 63.97%, and all compounds had a protective effect on neuronal cell damage induced by tert-butyl hydroperoxide. The experimental results of some compounds are shown in Table 5, and some compounds, such as compound ET16, had a better neuronal cell protective effect than edaravone.

[0181] Table 5 Protective effect of compounds on SH-SY5Y cell damage induced by tBHP

[0182]

[0183] Example 45: Catalase (CAT) and superoxide dismutase (SOD) activity test experiment

[0184] Thirty adult male ICR mice (8 weeks) were selected and randomly divided into 5 groups, namely: blank group, scopolamine group (film formation group), ET11 plus scopolamine group, ET21 plus scopolamine group, donepezil plus scopolamine group (positive drug group). The compound was prepared into an intraperitoneal injection solution using 2% DMSO, 48% PEG400, and 50% saline. The film formation group was administered at a dose of 1 mg / kg, and the compound group was administered at a dose of 5 mg / kg, and intraperitoneal injection was used. The activity of catalase (CAT) in brain tissue was determined using the ammonium molybdate detection method. After 1 hour of administration, the mice were euthanized, the brain tissue weight was accurately weighed, and nine times the volume of saline was added according to weight (g): volume (mL) = 1:9, and mechanical homogenization was performed at 4000r / min and centrifuged for ten minutes. CAT activity was determined according to the kit operation table. The total superoxide dismutase (T-SOD) enzyme activity was measured using the xanthine oxidase method. The sample pretreatment was the same as the CAT pretreatment. T-SOD activity was determined according to the kit operation table. The results showed (Table 6): The CAT enzyme activity of the groups using compounds ET11 and ET21 was higher than that of the scopolamine group, among which the CAT enzyme activity of the ET11 group was higher, close to that of the donepezil group. The T-SOD enzyme activity of the group using compound ET11 was higher than that of the scopolamine group, but the difference with the scopolamine group was not large, and the T-SOD enzyme activity of the ET11 group was slightly lower than that of the donepezil group.

[0185] Table 6 Effects of compounds on CAT and T-SOD

[0186]

[0187] Example 46: Metal Chelation Test

[0188] Taking compound ET11 as an example, this example uses ultraviolet spectroscopy to explore the neurotoxic metal ion chelating ability of the target compound. The metal salts used in the test are aluminum chloride, cuprous iodide, cupric chloride, ferrous sulfate, ferric chloride and zinc chloride. A methanol solution of the test compound (1.0 mL, 300 μM) was added to an aqueous solution containing metal ions (1.0 mL, 300 μM) and incubated on a shaker at 37°C for 1 hour. The solution was then transferred to a quartz cell to measure the absorbance of the solution at 200-550 nm. The experimental results are shown in Figure 2. Figure 2 As shown in a, compound ET11 and Cu 2+ with Fe 3+ When the compound coexists, the full wavelength changes significantly, indicating that the compound ET11 and Cu 2+ with Fe 3+ Therefore, this example follows the same determination method, keeping the concentration of compound ET11 unchanged (300 μM) and changing the concentration of Cu 2+The concentration of compound ET11 was determined by mixing with Cu 2+ The changes in the absorbance of the compounds at 200–550 nm when they coexist. Figure 2 As shown in b, when compound ET11 is combined with Cu 2+ When the compound coexists with Cu in a ratio of 2:1, 2+ The interaction approaches a steady state. 2+ The full wavelength of the compound did not change significantly. Therefore, compound ET11 may be related to Cu 2+ Chelation occurs in a quantitative ratio of 2:1.

[0189] Example 47: Water maze experiment

[0190] Thirty adult male ICR mice (8 weeks) were selected and randomly divided into 5 groups, namely: blank group, scopolamine group (film formation group), ET11 plus scopolamine group, ET21 plus scopolamine group, donepezil plus scopolamine group (positive drug group). The compound was prepared into an intraperitoneal injection solution using 2% DMSO, 48% PEG400, and 50% saline. The film formation group was administered at a dose of 1 mg / kg, and the compound group was administered at a dose of 5 mg / kg, and the administration was administered by intraperitoneal injection. The acquisition experiment was performed 5 days after administration. The mice were placed in the water maze laboratory one day before the experiment to adapt to the experimental environment. The mice were placed in the water from the set entry point facing the pool wall. The experimental mice climbed onto the platform within 90s and stayed for more than 3 seconds. It was considered to have found the platform. If the mouse still did not find the platform after more than 90s, the experimenter led the mouse to the platform. The time from entering the water to finding the platform was recorded as the escape latency. If the platform was not found within 90 seconds, the escape latency was recorded as 90 seconds. The training was conducted once a day, and the average value was taken as the escape latency. On the last day after all the training, the mice were subjected to an exploratory experiment. After the platform was removed, the mice were placed in the quadrant opposite to the quadrant where the platform was located with their heads facing the pool wall. The experimental animal behavior monitoring and analysis system was used to record the behavioral data such as the time required for the mice to successfully address (s), the addressing path, the number of quadrant crossings (n), and the swimming speed. Statistical analysis was performed to evaluate the cognitive ability of the mice. The results are shown in Figure 3 As shown, it can be seen that compared with the blank group, the addressing time of the model group mice is longer and the addressing path is disordered. The compounds E11 and ET21 of the present invention significantly shorten the addressing time of the model mice, and the addressing path is concise and clear.

[0191] Example 48: Immunohistochemistry test

[0192] After the water maze test, mice in each group were euthanized, and then the brains were fixed in 4% paraformaldehyde solution. The coronal sections of the brain were dewaxed and then stained with Nissl and glycine silver, respectively, and finally the stained sections were scanned using a slide scanner. The liver tissues of each group were stained with HE, and the liver tissue section samples were dewaxed, stained with hematoxylin, eosin, dehydrated and sealed, and finally photographed with a slide scanner. The results of Nissl staining, glycine silver staining, and H&E staining are shown in Figure 2. Figure 4 and 5 As shown, it can be seen that compounds ET11 and ET21 can improve the damage of neurons in the hippocampus of the mouse brain caused by scopolamine, and do not show obvious liver toxicity.

[0193] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

Claims

1. A pyrazolone-tacrine compound, characterized in that: It is a compound represented by formula (I), an enol tautomer of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof; Wherein, Z is selected from any of the following: X is a hydrogen or halogen atom; n is an integer of 1 to 3; and m is an integer of 1 to 2.

2. The pyrazolone-tacrine compound according to claim 1, characterized in that: The compound represented by formula (I) is selected from any one of the following ET1 to ET21:

3. A method for preparing the pyrazolone-tacrine compound according to any one of claims 1 to 2, characterized in that: The preparation method comprises: The compound represented by formula (1) is subjected to a cyclization reaction with dimethyl butynedioate to obtain a compound represented by formula (2); The compound represented by formula (2) is subjected to a hydrolysis reaction under alkaline conditions to obtain a compound represented by formula (3); The compound represented by formula (3) is subjected to condensation reaction with the compound represented by formula (4) to obtain the compound represented by formula (I); Wherein, the definitions of Z, X, n and m are as described in claim 1.

4. The preparation method according to claim 3, characterized in that: The molar ratio of the compound represented by formula (1) to dimethyl butynedioate is 1:0.9-1.

5. The conditions of the cyclization reaction include: reacting at 0°C for 2-4 hours in the presence of methanol, adding sodium methoxide to the reaction solution, and refluxing at 60-80°C for 0.5-2 hours.

5. The preparation method according to claim 3, characterized in that: The conditions of the hydrolysis reaction include: reflux at 30-50° C. for 4-6 hours in the presence of methanol, tetrahydrofuran and a base.

6. The preparation method according to claim 3, characterized in that: The molar ratio of the compound represented by formula (3) to the compound represented by formula (4) is 1:1.0-1.

5. The conditions of the condensation reaction include: reacting at room temperature for 20-30 hours under the protection of an inert atmosphere and in the presence of a solvent and a condensation agent.

7. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the pyrazolone-tacrine compound according to any one of claims 1 to 2.

8. The pharmaceutical composition according to claim 7, characterized in that The pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

9. The pharmaceutical composition according to claim 7, characterized in that The dosage form of the pharmaceutical composition is capsule, pill, tablet, granule or injection.

10. Use of the pyrazolone-tacrine compound according to any one of claims 1 to 2 in any one of the following i) to v): i) for preparing a drug for inhibiting acetylcholinesterase activity; ii) for preparing drugs for scavenging free radicals; iii) for preparing drugs for protecting neuronal cells; iv) for preparing a drug for improving animal memory and cognition; v) Used for preparing drugs for preventing, treating or assisting in treating neurodegenerative diseases.

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