Coumarin-quinoline derivatives, preparation methods and applications thereof

Abstract

The present solution discloses a coumarin-quinoline derivative in the field of medicinal chemistry, which combines a coumarin structure with anti-Aβ 42 protein aggregation activity and the pharmacophore quinoline ring of anti-Alzheimer's drugs, and adjusts the length between the two structures with an alkyl chain. Its structural formula is shown in Formula (I) or Formula (II), #imgabs0# The coumarin-quinoline derivative described in the present invention can be applied to the preparation of drugs for treating Alzheimer's disease, cerebrovascular dementia or myasthenia gravis, and the dosage form of the drug can be selected from tablets, pills, capsules, injections, suspensions or emulsions.

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and particularly relates to coumarin-quinoline derivatives, their preparation methods and applications. Background Art

[0002] Alzheimer's disease (AD), also known as senile dementia, is a chronic neurodegenerative disease. Senile dementia has become a highly prevalent disease causing death among the middle-aged and elderly, and it is also extremely costly. The global cost for treating and caring for Alzheimer's patients accounts for 11.2% of the total expenditure on diseases of the population aged 60 and above, far higher than that of stroke, cardiovascular diseases, and cancer, bringing a heavy burden to the families of patients and society. The aging population in China is increasing year by year. Currently, there are approximately 10 million AD patients in China, ranking first in the world. At the same time, China is also one of the countries with the fastest growth rate of the number of dementia patients globally. More seriously, for this neurodegenerative disease, no specific medicine or treatment method has been found yet, and only drugs can be used to relieve brain atrophy.

[0003] At present, the types and quantities of commonly used anti-AD drugs in clinical practice are relatively limited. They are mainly cholinesterase inhibitors or NMDA receptor antagonists. However, these drugs can only improve the cognitive level and quality of life of patients in the short term and cannot substantially reverse the degenerative process of nerve cells.

[0004] The causes of Alzheimer's disease are complex, including genetics, trauma, viral infections, etc. And its pathogenesis involves the regulation of multiple signaling pathways, such as the misfolding and aggregation of β-amyloid protein (Aβ protein), oxidative stress, inflammation, etc. And these factors interact and are interrelated with each other. Regarding complex diseases including AD, the internationally widely recognized concept of polypharmacology believes that the essence of the occurrence of complex diseases is the imbalance of the biological network caused by the combined action of multiple factors, rather than simply being caused by a single factor. The traditional "one drug, one target, one disease" treatment strategy is often difficult to meet the treatment requirements for complex diseases.

[0005] More and more studies have shown that drugs that act on multiple disease-related targets simultaneously, that is, multi-target drugs, may have better therapeutic effects. This has achieved certain success in the treatment of various diseases including AIDS, cancer, and depression. If a drug can simultaneously intervene in multiple links or multiple nodal proteins that induce the disease and regulate the entire disease mechanism network, it will exert a better therapeutic effect. For this reason, a multi-target anti-AD strategy has been proposed, that is, to design multi-target drugs that simultaneously contain multiple target pharmacophores of AD, act on multiple targets in the pathogenic mechanism of AD simultaneously, produce a synergistic effect, and thus achieve the best therapeutic effect. The multi-target anti-AD strategy conforms to the complex pathological conditions of AD and is a promising treatment strategy. Summary of the Invention

[0006] The present invention aims at the deficiencies of the prior art and provides a coumarin-quinoline derivative, a preparation method thereof and an application thereof.

[0007] In the coumarin-quinoline derivative in this solution, its structural formula is shown as formula (I) or formula (II),

[0008]

[0009] In formula (I), n is 2 or 3 or 4 or 5; R and R1 are hydrogen or methyl; R2 is hydrogen or chlorine; in formula (II), n is 2 or 3 or 4 or 5; R2 is hydrogen or chlorine.

[0010] Furthermore, the coumarin-quinoline derivative is selected from the following formula 3n or 3x:

[0011]

[0012] The present invention also provides a preparation method of the coumarin-quinoline derivative, which involves fusing the coumarin structure with anti-Aβ 42 protein aggregation activity and the pharmacophore quinoline ring of the anti-Alzheimer's disease drug, and simultaneously changing the length between the two structures.

[0013] Specifically: The preparation process of formula (I) is as follows:

[0014] (1) Dissolve and bromoalkane in solvent A, and carry out a substitution reaction under the action of acid-binding agent B to obtain compound The solvent A is one or several mixed reagents of acetone, dichloromethane, chloroform, acetonitrile, tetrahydrofuran, dioxane or dimethylformamide, preferably acetone;

[0015] (2) React with 5-chloro-8-hydroxyquinoline or 8-hydroxyquinoline to obtain the target product

[0016] The preparation process of formula (II) is as follows:

[0017] (1) Dissolve and bromoalkane in solvent A, and carry out a substitution reaction under the action of acid-binding agent B to obtain compound The solvent A is one or several mixed reagents of acetone, dichloromethane, chloroform, acetonitrile, tetrahydrofuran, dioxane or dimethylformamide, preferably acetone;

[0018] (2) React with 5-chloro-8-hydroxyquinoline or 8-hydroxyquinoline to obtain the target product

[0019] Optimized, the obtained target product was purified by column chromatography.

[0020] Furthermore, the bromoalkane is 1,2-dibromoethane, 1,3-dibromopropane, 1,4-dibromobutane or 1,5-dibromopentane.

[0021] Furthermore, the acid-binding agent B used is one or a mixture of reagents such as potassium carbonate, sodium carbonate, cesium carbonate, triethylamine, and N,N-diisopropylethylamine.

[0022] The coumarin-quinoline derivative described in the present invention can be applied to the preparation of drugs for treating Alzheimer's disease, cerebrovascular dementia or myasthenia gravis, and the drug dosage form can be selected from tablets, pills, capsules, injections, suspensions or emulsions.

[0023] The designed and synthesized coumarin-quinoline derivative of the present invention has good inhibition of Aβ 42 protein aggregation, inhibits the activities of monoamine oxidase and butyrylcholinesterase, and can penetrate the blood-brain barrier. Most of the compounds have an Aβ 42 protein aggregation rate of more than 40% at a concentration of 10 μM.

[0024] Among them, the inhibition rate of compound 3x on Aβ 42 protein self-aggregation reaches 61.2%, which is comparable to the positive controls curcumin and resveratrol; the IC 50 of compound 3n for inhibiting monoamine oxidase B reaches 0.77 μM, which is significantly better than the positive control ladostigil.

[0025] Furthermore, the IC 50 of compound 3x for inhibiting butyrylcholinesterase reaches 0.15 μM, and it can penetrate the blood-brain barrier, with a Pe value of 12.4, and can be used as a potential multi-functional anti-Alzheimer's drug. At the same time, 3x can inhibit Aβ 42 protein self-aggregation, induce SYSH-5Y (neuroblastoma) cell toxicity, and thus protect nerve cells; in addition, at the animal level, 3x can improve the cognitive impairment of APP / PS1 ((Alzheimer's disease)-neurodegenerative disease model) double transgenic mice.

[0026] The coumarin-quinoline derivative provided by the present invention has the activity of inhibiting Aβ 42 protein self-aggregation, inhibiting the activities of monoamine oxidase and butyrylcholinesterase, and can penetrate the blood-brain barrier. It is particularly suitable, but not limited to, the preparation of drugs for treating Alzheimer's disease, cerebrovascular dementia or myasthenia gravis.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] 1. The present invention provides coumarin - quinoline derivatives, and the preparation method of the derivatives is simple and easy to synthesize.

[0029] 2. Experiments show that the derivatives of the present invention have the activity of inhibiting the self - aggregation of Aβ 42 protein, inhibiting the activities of acetylcholinesterase, butyrylcholinesterase, and monoamine oxidase related to the onset of AD, and can cross the blood - brain barrier, having high medical value and broad market development prospects. Brief Description of the Drawings

[0030] Figure 1 Compound 3x in the present invention protects SYSH - 5Y cells from Aβ 42 protein - induced toxicity;

[0031] Figure 2 Compound 3x in the present invention improves the cognitive impairment of APP / PS1 transgenic mice. Detailed Embodiments

[0032] The technical solutions of the present invention are further described below through specific embodiments.

[0033] Unless otherwise specified, the reagents, equipment, and methods used in the present invention are conventional commercially available reagents, equipment, and methods commonly used in the technical field.

[0034] Taking the reaction in the presence of potassium carbonate and acetone during the process of obtaining the intermediate as an example, when preparing the structural formula (I), its synthesis process is as follows:

[0035]

[0036] Taking the reaction in the presence of potassium carbonate and acetone during the process of obtaining the intermediate as an example, when preparing the structural formula (II), its synthesis process is as follows:

[0037]

[0038] Example 1: Synthesis of Intermediate 2a

[0039] Add 7 - hydroxycoumarin (2 g, 12.34 mmol), K2CO3 (5.11 g, 37 mmol), 1,2 - dibromoethane (6.38 mL, 74.04 mmol) and 20 mL of acetone into a 100 mL flask for reflux reaction, and monitor the progress of the reaction by TLC. After the reaction is completed, rotary evaporate the acetone under reduced pressure, add 50 mL of distilled water, stir well, filter, dry the filter cake, and then separate and purify by silica gel column chromatography to obtain 2.6 g of white solid. The yield is 78.33%.

[0040]

[0041] Example 2: Synthesis of Intermediate 2b

[0042] The synthesis method is the same as that of 2a, using 1,3-dibromopropane instead of 1,2-dibromoethane. 2.64 g of white solid was obtained by silica gel column chromatography, and the yield was 75.6%.

[0043]

[0044] Example 3: Synthesis of Intermediate 2c

[0045] The synthesis method is the same as that of 2a, using 1,4-dibromobutane instead of 1,2-dibromoethane; using dichloromethane instead of acetone; using sodium carbonate instead of potassium carbonate. 2.58 g of white solid was obtained by silica gel column chromatography, and the yield was 70.4%.

[0046]

[0047] Example 4: Synthesis of Intermediate 2d

[0048] The synthesis method is the same as that of 2a, using 1,5-dibromopentane instead of 1,2-dibromoethane; using chloroform instead of acetone; using cesium carbonate instead of potassium carbonate. 3.01 g of white solid was obtained by silica gel column chromatography, and the yield was 78.4%.

[0049]

[0050] Example 5: Synthesis of Intermediate 2e

[0051] The synthesis method is the same as that of 2a, using 1,5-dibromopentane instead of 1,2-dibromoethane; using acetonitrile instead of acetone; using triethylamine instead of potassium carbonate. 2.85 g of white solid was obtained by silica gel column chromatography, and the yield was 88.7%.

[0052]

[0053] Example 6: Synthesis of Intermediate 2f

[0054] The synthesis method is the same as that of 2a, using 4-methylumbelliferone instead of 7-hydroxycoumarin; using tetrahydrofuran instead of acetone; using N,N-diisopropylethylamine instead of potassium carbonate, and 1,3-dibromopropane instead of 1,2-dibromoethane. 2.1 g of white solid was obtained by silica gel column chromatography, and the yield was 62.3%.

[0055]

[0056] Example 7: Synthesis of Intermediate 2g

[0057] The synthesis method was the same as that of 2a, using 4-methylumbelliferone instead of 7-hydroxycoumarin; using tetrahydrofuran instead of acetone; using sodium carbonate instead of potassium carbonate, and 1,4-dibromobutane instead of 1,2-dibromoethane. A white solid of 2.32 g was obtained by silica gel column chromatography, with a yield of 65.7%.

[0058]

[0059] Example 8: Synthesis of Intermediate 2h

[0060] The synthesis method was the same as that of 2a, using 4-methylumbelliferone instead of 7-hydroxycoumarin; using tetrahydrofuran instead of acetone, and 1,4-dibromopentane instead of 1,2-dibromoethane. A white solid of 2.93 g was obtained by silica gel column chromatography, with a yield of 79.4%.

[0061]

[0062] Example 9: Synthesis of Intermediate 2i

[0063] The synthesis method was the same as that of 2a, using 4-hydroxycoumarin instead of 7-hydroxycoumarin; using acetonitrile instead of acetone. A white solid of 2.15 g was obtained by silica gel column chromatography, with a yield of 64.77%.

[0064]

[0065] Example 10: Synthesis of Intermediate 2j

[0066] The synthesis method was the same as that of 2a, using 4-hydroxycoumarin instead of 7-hydroxycoumarin; using acetonitrile instead of acetone, and 1,3-dibromopropane instead of 1,2-dibromoethane. A white solid of 2.23 g was obtained by silica gel column chromatography, with a yield of 63.8%.

[0067]

[0068] Example 11: Synthesis of Intermediate 2k

[0069] The synthesis method was the same as that of 2a, using 4-hydroxycoumarin instead of 7-hydroxycoumarin, and 1,4-dibromobutane instead of 1,2-dibromoethane.

[0070] A white solid of 2.58 g was obtained by silica gel column chromatography, with a yield of 70.4%.

[0071]

[0072] Example 12: Synthesis of Intermediate 2l

[0073] The synthesis method was the same as that of 2a, using 4-hydroxycoumarin instead of 7-hydroxycoumarin, 1,3-dibromopentane instead of 1,2-dibromoethane; using cesium carbonate instead of potassium carbonate. A white solid of 2.18 g was obtained by silica gel column chromatography with a yield of 56.8%.

[0074]

[0075] Example 13: Synthesis of Compound 3a

[0076] 5-Chloro-8-hydroxyquinoline (0.13 g, 0.74 mmol), K2CO3 (0.20 g, 1.48 mmol) and 10 mL of acetonitrile were added to a 50 mL flask and refluxed. After 30 min, the substituted coumarin intermediate 2a (0.20 g, 0.74 mmol) was added, and the reflux reaction was continued. The progress of the reaction was monitored by TLC. After the reaction was completed, the acetonitrile was evaporated under reduced pressure, 50 mL of water was added, and the mixture was stirred thoroughly, filtered by suction, and the filter cake was dried. Then it was separated and purified by silica gel column chromatography to obtain 0.20 g of a white solid. The yield was 72.5%. 1 H NMR (500 MHz, CDCl3) δ 9.01 (dd, J = 4.2, 1.7 Hz, 1H), 8.55 (dd, J = 8.5, 1.7 Hz, 1H), 7.64 (d, J = 9.5 Hz, 1H), 7.60–7.47 (m, 2H), 7.41–7.35 (m, 1H), 7.10 (d, J = 8.4 Hz, 1H), 6.96–6.88 (m, 2H), 6.27 (d, J = 9.5 Hz, 1H), 4.65 (dd, J = 5.6, 3.5 Hz, 2H), 4.60 (dd, J = 5.6, 3.5 Hz, 2H). 13 C NMR (126 MHz, CDCl3) δ 161.67, 161.20, 155.76, 153.54, 149.97, 143.40, 140.76, 133.13, 128.85, 127.21, 126.33, 123.11, 122.52, 113.37, 112.98, 112.89, 109.42, 101.78, 67.41, 66.89.

[0077]

[0078] Example 14: Synthesis of Compound 3b

[0079] The synthesis method was the same as that of 3a, except that the coumarin intermediate 2b was used instead of 2a. A white solid of 0.23 g was obtained by column chromatography separation and purification with a yield of 83.6%. 11H NMR (500 MHz, CDCl3) δ 9.00 (dd, J = 4.2, 1.7 Hz, 1H), 8.54 (dd, J = 8.5, 1.7 Hz, 1H), 7.62 (d, J = 9.4 Hz, 1H), 7.59–7.50 (m, 2H), 7.39–7.32 (m, 1H), 7.04 (d, J = 8.4 Hz, 1H), 6.89–6.83 (m, 2H), 6.24 (d, J = 9.4 Hz, 1H), 4.80–4.76 (m, 1H), 4.45 (t, J = 6.2 Hz, 2H), 4.34 (t, J = 5.9 Hz, 2H), 2.54 (p, J = 6.1 Hz, 2H), 1.32–1.24 (m, 1H). 13 13C NMR (126 MHz, CDCl3) δ 162.06, 161.27, 155.82, 153.81, 149.84, 143.43, 140.79, 133.06, 128.75, 127.13, 126.40, 122.44, 122.42, 113.11, 112.80, 112.60, 108.82, 101.63, 65.49, 65.18, 28.86.

[0080]

[0081] Example 15: Synthesis of Compound 3c

[0082] The synthesis method was the same as that of 3a, except that coumarin intermediate 2c was used instead of 2a. 0.23 g of white solid was obtained by column chromatography separation and purification, and the yield was 88.6%.

[0083]

[0084] Example 16: Synthesis of Compound 3d

[0085] The synthesis method was the same as that of 3a, except that coumarin intermediate 2d was used instead of 2a. 0.23 g of white solid was obtained by column chromatography separation and purification, and the yield was 87.6%. 11H NMR (500 MHz, CDCl3) δ 9.00 (dd, J = 4.2, 1.7 Hz, 1H), 8.54 (dd, J = 8.6, 1.7 Hz, 1H), 7.64 (d, J = 9.4 Hz, 1H), 7.59–7.50 (m, 2H), 7.35 (d, J = 8.6 Hz, 1H), 6.99 (d, J = 8.4 Hz, 1H), 6.83 (dd, J = 8.5, 2.4 Hz, 1H), 6.80 (d, J = 2.4 Hz, 1H), 6.25 (d, J = 9.5 Hz, 1H), 4.27 (t, J = 6.7 Hz, 2H), 4.07 (t, J = 6.3 Hz, 2H), 2.12 (p, J = 6.9 Hz, 2H), 2.00–1.91 (m, 2H), 1.82–1.72 (m, 2H). 13 13C NMR (126 MHz, CDCl3) δ 162.29, 161.35, 155.87, 153.98, 149.78, 143.50, 140.84, 133.01, 128.73, 127.11, 126.43, 122.34, 122.07, 112.96, 112.91, 112.44, 108.54, 101.36, 68.91, 68.33, 28.78, 28.65, 22.73.

[0086]

[0087] Example 17: Synthesis of Compound 3e

[0088] The synthesis method was the same as that of 3a, except that coumarin intermediate 2e was used instead of 2a. 0.20 g of white solid was obtained by column chromatography separation and purification, and the yield was 73.8%. 1 1H NMR (500 MHz, CDCl3) δ 9.03 (dd, J = 4.2, 1.7 Hz, 1H), 8.56 (dd, J = 8.5, 1.7 Hz, 1H), 7.56 (td, J = 8.5, 4.7 Hz, 2H), 7.50 (dd, J = 8.5, 2.4 Hz, 1H), 7.17–7.09 (m, 1H), 6.96–6.89 (m, 2H), 6.18–6.14 (m, 1H), 4.65 (dd, J = 5.8, 3.6 Hz, 2H), 4.61 (dt, J = 5.7, 2.6 Hz, 2H), 2.40 (d, J = 1.4 Hz, 3H). 1313C NMR (126 MHz, CDCl3) δ 161.53, 161.26, 155.17, 153.58, 152.52, 150.00, 140.79, 133.11, 127.21, 126.33, 125.60, 123.10, 122.51, 113.92, 112.62, 112.20, 109.45, 101.79, 67.46, 66.83.

[0089]

[0090] Example 18: Synthesis of Compound 3f

[0091] The synthesis method was the same as that of 3a, except that coumarin intermediate 2f was used instead of 2a. 0.25 g of white solid was obtained by column chromatography separation and purification, and the yield was 93.2%. 1 1H NMR (500 MHz, CDCl3) δ 9.02 (dt, J = 3.0, 1.6 Hz, 1H), 8.54 (ddt, J = 7.2, 3.4, 1.6 Hz, 1H), 7.55 (dddd, J = 18.0, 9.7, 3.6, 1.4 Hz, 2H), 7.47 (dt, J = 8.7, 2.0 Hz, 1H), 7.04 (dt, J = 8.4, 1.6 Hz, 1H), 6.91–6.83 (m, 2H), 6.13 (q, J = 1.7 Hz, 1H), 4.49–4.43 (m, 2H), 4.34 (t, J = 5.9 Hz, 2H), 2.54 (p, J = 6.1 Hz, 2H), 2.39 (dt, J = 2.5, 1.5 Hz, 3H). 13 13C NMR (126 MHz, CDCl3) δ 161.87, 161.31, 155.21, 153.84, 152.51, 149.87, 140.83, 133.03, 127.13, 126.38,

[0092] 125.51, 122.43, 122.41, 113.65, 112.44, 112.01, 108.83, 101.66, 65.53, 65.13, 28.88.

[0093]

[0094] Example 19: Synthesis of Compound 3g

[0095] The synthesis method was the same as that of 3a, except that coumarin intermediate 2g was used instead of 2a. 0.22 g of white solid was obtained by column chromatography separation and purification, and the yield was 82.1%. 11H NMR (500 MHz, CDCl3) δ 9.01 (dd, J = 4.2, 1.7 Hz, 1H), 8.51 (dd, J = 8.5, 1.7 Hz, 1H), 7.59–7.48 (m, 2H), 7.44 (d, J = 9.1 Hz, 1H), 6.99 (d, J = 8.4 Hz, 1H), 6.81 (d, J = 7.9 Hz, 2H), 6.13 (d, J = 1.4 Hz, 1H), 4.34 (t, J = 6.4 Hz, 2H), 4.18 (t, J = 6.2 Hz, 2H), 2.39 (d, J = 1.2 Hz, 3H), 2.24 (dq, J = 7.8, 6.4 Hz, 2H), 2.13 (dt, J = 8.6, 6.1 Hz, 2H). 13 13C NMR (126 MHz, CDCl3) δ 161.99, 161.38, 155.20, 153.82, 152.61, 149.84, 140.83, 132.91, 127.08, 126.38, 125.41, 122.35, 122.14, 113.43, 112.66, 111.84, 108.53, 101.30, 68.70, 68.17, 26.05, 25.49, 18.68.

[0096]

[0097] Example 20: Synthesis of Compound 3h

[0098] The synthesis method was the same as that of 3a, except that coumarin intermediate 2h was used instead of 2a. 0.22 g of white solid was obtained by column chromatography separation and purification, with a yield of 82.7%. 1 1H NMR (500 MHz, CDCl3) δ 9.01 (dd, J = 4.2, 1.6 Hz, 1H), 8.54 (dd, J = 8.5, 1.7 Hz, 1H), 7.59–7.50 (m, 2H), 7.47 (d, J = 8.8 Hz, 1H), 6.99 (d, J = 8.4 Hz, 1H), 6.84 (dd, J = 8.8, 2.5 Hz, 1H), 6.80 (d, J = 2.5 Hz, 1H), 6.13 (q, J = 1.3 Hz, 1H), 4.28 (t, J = 6.7 Hz, 2H), 4.07 (t, J = 6.3 Hz, 2H), 2.40 (d, J = 1.3 Hz, 3H), 2.17–2.08 (m, 2H), 1.95 (dt, J = 14.8, 6.5 Hz, 2H), 1.82–1.71 (m, 2H). 1313C NMR (126 MHz, CDCl3) δ 162.09, 161.39, 155.26, 153.99, 152.61, 149.80, 140.86, 133.00, 127.11, 126.43, 125.49, 122.33, 122.06, 113.47, 112.57, 111.86, 108.54, 101.38, 68.93, 68.27, 28.80, 28.65, 22.74, 18.69.

[0099]

[0100] Example 21: Synthesis of Compound 3i

[0101] The synthesis method was the same as that of 3a, except that coumarin intermediate 2i was used instead of 2a. 0.21 g of yellow solid was obtained by column chromatography separation and purification, and the yield was 78.6%. 1 1H NMR (500 MHz, CDCl3) δ 9.02 (dd, J = 4.2, 1.7 Hz, 1H), 8.57 (dd, J = 8.6, 1.6 Hz, 1H), 7.77 (dd, J = 8.0, 1.6 Hz, 1H), 7.56 (dddd, J = 19.1, 8.7, 6.7, 1.9 Hz, 3H), 7.34–7.26 (m, 1H), 7.26–7.19 (m, 1H), 7.15 (d, J = 8.4 Hz, 1H), 5.79 (s, 1H), 4.76 (dd, J = 5.8, 3.5 Hz, 2H), 4.70 (dd, J = 5.7, 3.5 Hz, 2H). 13 13C NMR (126 MHz, CDCl3) δ 165.39, 162.74, 153.53, 153.30, 150.09, 140.91, 133.18, 132.52, 127.32, 126.27, 123.88, 123.59, 123.24, 122.59, 116.74, 115.42, 110.13, 90.99, 67.65, 67.15.

[0102]

[0103] Example 22: Synthesis of Compound 3j

[0104] The synthesis method was the same as that of 3a, except that coumarin intermediate 2j was used instead of 2a. 0.10 g of light yellow solid was obtained by column chromatography separation and purification, and the yield was 35.7%. 11H NMR (500 MHz, CDCl3) δ 9.01 (dd, J = 4.2, 1.7 Hz, 1H), 8.54 (dd, J = 8.5, 1.7 Hz, 1H), 7.81 (dd, J = 7.9, 1.6 Hz, 1H), 7.60–7.50 (m, 3H), 7.31 (d, J = 8.3 Hz, 1H), 7.28–7.21 (m, 1H), 7.04 (d, J = 8.4 Hz, 1H), 5.75 (s, 1H), 4.48 (dt, J = 11.6, 6.0 Hz, 4H), 2.64 (p, J = 6.0 Hz, 2H). 13 13C NMR (126 MHz, CDCl3) δ 165.44, 162.84, 153.67, 153.30, 149.96, 140.81, 133.06, 132.40, 127.19, 126.32, 123.86, 122.90, 122.74, 122.50, 116.81, 115.62, 108.92, 90.81, 66.13, 65.37, 28.46, 0.02.

[0105]

[0106] Example 23: Synthesis of Compound 3k

[0107] The synthesis method was the same as that of 3a, except that coumarin intermediate 2k was used instead of 2a. After separation and purification by column chromatography, 0.24 g of a light yellow solid was obtained, with a yield of 89.7%. 1 1H NMR (500 MHz, CDCl3) δ 9.04 (dd, J = 4.2, 1.7 Hz, 1H), 8.52 (dd, J = 8.5, 1.7 Hz, 1H), 7.73 (dd, J = 8.0, 1.6 Hz, 1H), 7.60–7.47 (m, 3H), 7.32–7.27 (m, 1H), 7.25–7.18 (m, 1H), 6.99 (d, J = 8.4 Hz, 1H), 5.76 (s, 1H), 4.36 (dt, J = 8.0, 5.7 Hz, 4H), 2.26 (ttd, J = 16.4, 7.5, 4.7 Hz, 4H). 13 13C NMR (126 MHz, CDCl3) δ 165.54, 163.01, 153.68, 153.26, 149.98, 140.77, 132.96, 132.28, 127.10, 126.31, 123.78, 122.89, 122.52, 122.34, 116.72, 115.63, 108.50, 90.65, 69.17, 68.59, 25.92, 25.38.

[0108]

[0109] Example 24: Synthesis of Compound 3l

[0110] The synthesis method was the same as that of 3a, except that coumarin intermediate 2l was used instead of 2a. 0.16 g of white solid was obtained by column chromatography separation and purification, with a yield of 60.2%. 1 H NMR(500MHz,CDCl3)δ8.99(dd,J=4.2,1.7Hz,1H),8.54(dd,J=8.5,1.7Hz,1H),7.81(dd,J=7.9,1.7Hz,1H),7.58–7.50(m,3H),7.31(dd,J=8.4,1.1Hz,1H),7.24(ddd,J=8.1,7.4,1.1Hz,1H),6.99(d,J=8.4Hz,1H),5.67(s,1H),4.29(t,J=6.6Hz,2H),4.18(t,J=6.3Hz,2H),2.15(dt,J=14.4,6.8Hz,2H),2.04(dq,J=8.2,6.4Hz,2H),1.87–1.77(m,2H). 13 C NMR(126MHz,CDCl3)δ165.65,163.00,153.93,153.32,149.83,140.87,133.00,132.35,127.12,126.40,123.83,123.01,122.35,122.16,116.77,115.74,108.56,90.43,69.13,68.77,28.59,28.34,22.79.

[0111]

[0112] Example 25: Synthesis of Compound 3m

[0113] The synthesis method was the same as that of 3a, except that 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. 0.19 g of white solid was obtained by column chromatography separation and purification, with a yield of 78.0%. 11H NMR (500 MHz, CDCl3) δ 8.99 (dd, J = 4.2, 1.8 Hz, 1H), 8.16 (dd, J = 8.2, 1.8 Hz, 1H), 7.65 (d, J = 9.5 Hz, 1H), 7.52–7.44 (m, 3H), 7.41–7.35 (m, 1H), 7.19 (dd, J = 7.3, 1.6 Hz, 1H), 6.92 (d, J = 7.6 Hz, 2H), 6.27 (d, J = 9.5 Hz, 1H), 4.67 (dd, J = 5.9, 4.0 Hz, 2H), 4.61 (dd, J = 5.8, 4.0 Hz, 2H). 13 13C NMR (126 MHz, CDCl3) δ 161.81, 161.21, 155.79, 154.29, 149.54, 143.41, 140.25, 136.09, 129.61, 128.82, 126.61, 121.78, 120.54, 113.31, 112.96, 112.82, 109.50, 101.80, 67.11, 66.95.

[0114]

[0115] Example 26: Synthesis of Compound 3n

[0116] The synthesis method was the same as that of 3a, except that coumarin intermediate 2b was used instead of 2a, and 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. After separation and purification by column chromatography, 0.19 g of white solid was obtained with a yield of 78.0%. 1 1H NMR (500 MHz, CDCl3) δ 8.96 (dd, J = 4.2, 1.7 Hz, 1H), 8.13 (dd, J = 8.3, 1.8 Hz, 1H), 7.61 (d, J = 9.5 Hz, 1H), 7.47–7.32 (m, 4H), 7.11 (dd, J = 7.6, 1.3 Hz, 1H), 6.88–6.82 (m, 2H), 6.23 (d, J = 9.4 Hz, 1H), 4.46 (t, J = 6.2 Hz, 2H), 4.33 (t, J = 6.0 Hz, 2H), 2.54 (p, J = 6.1 Hz, 2H). 13 13C NMR (126 MHz, CDCl3) δ 162.12, 161.26, 155.83, 154.57, 149.43, 143.43, 140.33, 135.98, 129.54, 128.74, 126.65, 121.67, 119.91, 113.08, 112.77, 112.56, 108.92, 101.67, 65.33, 65.21, 28.92.

[0117]

[0118] Example 27: Synthesis of Compound 3o

[0119] The synthesis method was the same as that of 3a, except that coumarin intermediate 2c was used instead of 2a, and 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. 0.22 g of white solid was obtained by column chromatography separation and purification, and the yield was 90.4%. 1 H NMR(500MHz,CDCl3)δ8.97(dd,J=4.2,1.7Hz,1H),8.13(dd,J=8.3,1.8Hz,1H),7.62(d,J=9.4Hz,1H),7.50–7.31(m,4H),7.08(dd,J=7.7,1.3Hz,1H),6.83(dq,J=4.3,2.4Hz,2H),6.25(d,J=9.5Hz,1H),4.35(t,J=6.4Hz,2H),4.19(t,J=6.3Hz,2H),2.25(dq,J=8.1,6.5Hz,2H),2.13(dq,J=9.3,6.4Hz,2H). 13 C NMR(126MHz,CDCl3)δ162.28,161.31,155.87,154.62,149.43,143.47,140.37,135.91,129.52,128.68,126.65,121.65,119.68,112.95,112.93,112.41,108.66,101.42,68.42,68.32,26.12,25.61.

[0120]

[0121] Example 28: Synthesis of Compound 3p

[0122] The synthesis method was the same as that of 3a, except that coumarin intermediate 2d was used instead of 2a, and 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. 0.22 g of white solid was obtained by column chromatography separation and purification, and the yield was 92.5%. 11H NMR (500 MHz, CDCl3) δ 8.95 (dt, J = 3.1, 1.5 Hz, 1H), 8.12 (dd, J = 8.2, 1.8 Hz, 1H), 7.61 (d, J = 9.4 Hz, 1H), 7.49–7.30 (m, 4H), 7.07 (dd, J = 7.7, 1.2 Hz, 1H), 6.85–6.74 (m, 2H), 6.23 (d, J = 9.4 Hz, 1H), 4.28 (t, J = 6.7 Hz, 2H), 4.05 (t, J = 6.4 Hz, 2H), 2.12 (p, J = 7.0 Hz, 2H), 1.94 (p, J = 6.7 Hz, 2H), 1.81–1.71 (m, 2H). 13 13C NMR (126 MHz, CDCl3) δ 162.32, 161.31, 155.87, 154.73, 149.35, 143.49, 140.35, 135.95, 129.52, 128.76, 128.74, 126.69, 121.58, 119.56, 112.92, 112.88, 112.41, 108.65, 101.37, 68.64, 68.37, 28.82, 28.74, 22.76.

[0123]

[0124] Example 29: Synthesis of Compound 3q

[0125] The synthesis method was the same as that of 3a, except that coumarin intermediate 2e was used instead of 2a, and 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. After separation and purification by column chromatography, 0.20 g of white solid was obtained with a yield of 80.7%. 1 1H NMR (500 MHz, CDCl3) δ 8.99 (dd, J = 4.2, 1.7 Hz, 1H), 8.17 (dd, J = 8.3, 1.7 Hz, 1H), 7.58–7.44 (m, 5H), 6.97–6.90 (m, 2H), 6.16 (q, J = 1.2 Hz, 1H), 4.70–4.65 (m, 2H), 4.65–4.59 (m, 2H), 2.41 (d, J = 1.2 Hz, 3H). 13 13C NMR (126 MHz, CDCl3) δ 161.62, 161.33, 155.18, 154.27, 152.57, 149.50, 136.15, 129.62, 126.64, 125.61, 124.48, 124.00, 121.80, 120.51, 112.65, 112.15, 109.46, 101.80, 67.10, 66.88, 18.71.

[0126]

[0127] Example 30: Synthesis of Compound 3r

[0128] The synthesis method was the same as that of 3a, except that coumarin intermediate 2f was used instead of 2a, and 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. After separation and purification by column chromatography, 0.17 g of white solid was obtained, and the yield was 68.4%. 1 H NMR(500MHz,CDCl3)δ8.97(dd,J=4.2,1.7Hz,1H),8.14(dd,J=8.3,1.8Hz,1H),7.49–7.39(m,4H),7.14(ddd,J=7.6,6.2,1.9Hz,1H),6.92–6.84(m,2H),6.13(q,J=1.3Hz,1H),4.48(t,J=6.2Hz,2H),4.35(t,J=6.0Hz,2H),2.55(p,J=6.1Hz,2H),2.39(d,J=1.2Hz,3H). 13 C NMR(126MHz,CDCl3)δ161.94,161.35,155.22,154.58,152.54,149.43,140.33,135.97,129.54,126.65,125.50,121.67,119.90,113.61,112.43,111.96,108.92,101.71,65.28,65.24,28.94,18.69.

[0129]

[0130] Example 31: Synthesis of Compound 3s

[0131] The synthesis method was the same as that of 3a, except that coumarin intermediate 2g was used instead of 2a, and 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. After separation and purification by column chromatography, 0.20 g of white solid was obtained, and the yield was 83.9%. 11H NMR (500 MHz, CDCl3) δ 8.98 (dq, J = 5.2, 2.2 Hz, 1H), 8.14 (dt, J = 8.4, 2.1 Hz, 1H), 7.51–7.35 (m, 4H), 7.18–7.06 (m, 1H), 6.85 (ddt, J = 9.5, 5.0, 2.4 Hz, 2H), 6.17–6.12 (m, 1H), 4.36 (td, J = 6.3, 2.7 Hz, 2H), 4.20 (td, J = 6.3, 2.6 Hz, 2H), 2.26 (ddt, J = 9.4, 6.8, 4.8 Hz, 2H), 2.15 (dt, J = 9.3, 6.2 Hz, 2H). 13 13C NMR (126 MHz, CDCl3) δ 162.09, 161.44, 155.26, 154.62, 152.62, 149.41, 135.94, 129.53, 126.67, 125.45, 121.65, 119.66, 113.46, 112.65, 111.86 (2C), 108.66, 101.44, 68.41, 68.26, 26.09, 25.65, 18.71.

[0132]

[0133] Example 32: Synthesis of Compound 3t

[0134] The synthesis method was the same as that of 3a, except that the coumarin intermediate 2h was used instead of 2a, and 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. After separation and purification by column chromatography, 0.18 g of light yellow solid was obtained with a yield of 73.4%. 1 1H NMR (500 MHz, CDCl3) δ 8.97 (dq, J = 4.2, 1.4 Hz, 1H), 8.15 (dq, J = 8.3, 1.7 Hz, 1H), 7.53–7.35 (m, 4H), 7.09 (dt, J = 7.7, 1.3 Hz, 1H), 6.91–6.78 (m, 2H), 6.14 (q, J = 1.5 Hz, 1H), 4.30 (td, J = 6.9, 1.4 Hz, 2H), 4.08 (tt, J = 6.4, 1.6 Hz, 2H), 2.41 (dt, J = 3.1, 1.3 Hz, 3H), 2.20–2.09 (m, 2H), 2.01–1.90 (m, 2H), 1.83–1.73 (m, 3H). 1313C NMR (126 MHz, CDCl3) δ 162.14, 161.42, 155.28, 154.76, 152.62, 149.38, 140.39, 135.94, 129.53, 126.68, 125.49, 121.59, 119.57, 113.47, 112.59, 111.86, 108.65, 101.41, 68.66, 68.34, 28.85, 28.75, 22.78, 18.71.

[0135]

[0136] Example 33: Synthesis of Compound 3u

[0137] The synthesis method was the same as that of 3a, except that coumarin intermediate 2i was used instead of 2a, and 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. After separation and purification by column chromatography, 0.09 g of a light yellow solid was obtained, with a yield of 36.1%. 1 1H NMR (500 MHz, CDCl3) δ 8.97 (dd, J = 4.2, 1.7 Hz, 1H), 8.17 (dd, J = 8.3, 1.8 Hz, 1H), 7.80 (dd, J = 7.9, 1.6 Hz, 1H), 7.56–7.42 (m, 4H), 7.33–7.28 (m, 1H), 7.24–7.19 (m, 2H), 5.78 (s, 1H), 4.77 (dd, J = 5.8, 3.7 Hz, 2H), 4.70 (dd, J = 5.8, 3.7 Hz, 2H). 13 13C NMR (126 MHz, CDCl3) δ 165.46, 162.78, 154.26, 153.29, 149.62, 140.39, 136.08, 132.47, 129.69, 126.53, 123.85, 123.34, 121.84, 120.96, 116.69, 115.48, 110.11, 90.92, 67.74, 66.79.

[0138]

[0139] Example 34: Synthesis of Compound 3v

[0140] The synthesis method was the same as that of 3a, except that coumarin intermediate 2j was used instead of 2a, and 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. After separation and purification by column chromatography, 0.15 g of a white solid was obtained, with a yield of 60.4%. 11H NMR (500 MHz, CDCl3) δ 8.98 (dd, J = 4.1, 1.7 Hz, 1H), 8.15 (dd, J = 8.3, 1.7 Hz, 1H), 7.83 (dd, J = 7.9, 1.6 Hz, 1H), 7.55 (ddd, J = 8.8, 7.4, 1.7 Hz, 1H), 7.50–7.40 (m, 3H), 7.32 (d, J = 8.3 Hz, 1H), 7.29–7.22 (m, 1H), 7.13 (dd, J = 7.4, 1.5 Hz, 1H), 5.75 (s, 1H), 4.51 (t, J = 6.1 Hz, 2H), 4.48 (t, J = 6.0 Hz, 2H), 2.65 (p, J = 6.1 Hz, 2H). 13 13C NMR (126 MHz, CDCl3) δ 165.50, 162.89, 154.42, 153.32, 149.49, 140.27, 136.04, 132.37, 129.58, 126.60, 123.86, 122.97, 121.77, 120.17, 116.79, 115.68, 109.01, 90.79, 66.28, 65.08, 28.54.

[0141]

[0142] Example 35: Synthesis of Compound 3w

[0143] The synthesis method was the same as that of 3a, except that coumarin intermediate 2k was used instead of 2a, and 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. 0.18 g of white solid was obtained by column chromatography separation and purification, and the yield was 74.9%.

[0144]

[0145] Example 36: Synthesis of Compound 3x

[0146] The synthesis method was the same as that of 3a, except that coumarin intermediate 2l was used instead of 2a, and 8-hydroxyquinoline was used instead of 5-chloro-8-hydroxyquinoline. 0.14 g of white solid was obtained by column chromatography separation and purification, and the yield was 58.0%. 11H NMR (500 MHz, CDCl3) δ 8.96 (dd, J = 4.3, 1.8 Hz, 1H), 8.15 (dd, J = 8.3, 1.8 Hz, 1H), 7.83 (dd, J = 7.9, 1.6 Hz, 1H), 7.55 (ddd, J = 8.7, 7.2, 1.6 Hz, 1H), 7.52–7.35 (m, 3H), 7.33 (dd, J = 8.4, 1.0 Hz, 1H), 7.29–7.22 (m, 1H), 7.09 (dd, J = 7.6, 1.3 Hz, 1H), 5.68 (s, 1H), 4.32 (t, J = 6.6 Hz, 2H), 4.19 (t, J = 6.3 Hz, 2H), 2.17 (p, J = 6.8 Hz, 2H), 2.11–2.01 (m, 2H), 1.89–1.79 (m, 2H). 13 13C NMR (126 MHz, CDCl3) δ 165.69, 163.05, 154.69, 153.34, 149.38, 140.36, 135.98, 132.34, 129.55, 126.66, 123.85, 123.07, 121.62, 119.66, 116.77, 115.78, 108.68, 90.43, 77.31, 77.05, 76.80, 69.18, 68.49, 28.66, 28.37, 22.82.

[0147]

[0148] Example 37: Inhibitory effect of the quinoline derivative of the present invention on Aβ self-aggregation:

[0149] Select the compounds prepared in Examples 13 to 34 and use the ThT method to measure the Aβ self-aggregation inhibitory activity.

[0150] 1. Preparation of solutions

[0151] (1) 10 mM pH 7.4 phosphate buffer solution (PBS): Take 3.618 g of Na2HPO4 and 0.603 g of KH2PO4 in a beaker, and make up to 100 mL with ultrapure water as the stock solution. Take 4 mL of the stock solution and dilute it to 100 mL with ultrapure water, adjust the pH to 7.4, and store it at 4 °C for later use.

[0152] (3) Aβ 42 Protein solution: Add 1 mg of protein to 500 μL of hexafluoroisopropanol, place it at room temperature overnight, wait for the protein to completely dissolve and make the aggregated Aβ 42Completely disperse and evenly aliquot into 10 200 μL EP tubes, freeze overnight at -80 °C, then lyophilize using a freeze dryer, and store the lyophilized powder at -20 °C for later use. When in use, dissolve it in DMSO to a concentration of 5 mM, and then dilute it with PBS buffer to 100 μM.

[0153] (4) 50 mM glycine-NaOH buffer: Weigh 0.938 g of glycine into a beaker, add 250 mL of ultrapure water to dissolve it completely, adjust the pH to 8.5 with NaOH solution, and store it at 4 °C for later use.

[0154] (5) 5 μM thioflavin T solution: Weigh 3.1 mg of ThT powder into a 1.5 mL EP tube, add 970 μL of glycine-NaOH solution to make its concentration 10 mM as a stock solution, and store it at -20 °C in the dark. Before use, take 3 μL of the stock solution and dilute it to 5 μM with 6 mL of glycine-NaOH solution. This solution is prepared and used immediately.

[0155] (7) 10.0 mM sample solution: Weigh a certain amount of each sample to be analyzed and dissolve it in an appropriate amount of DMSO to prepare a 10.0 mM sample solution, and then dilute it to 40 μM with 10 mM PBS at pH 7.4.

[0156] 2. Aβ 42 Protein self-aggregation inhibition activity test

[0157] Take 10 μL of 50 μM Aβ 1-42 protein and 10 μL of 20 μM compound solution in a 200 μL EP tube and incubate at 37 °C for 48 hours. 10 μL of protein and 10 μL of PBS are set as the blank control, and the positive controls are 10 μL of protein and 10 μL of curcumin or resveratrol or donepezil. After 48 hours, add 180 μL of 5 μM ThT solution to the 200 μL EP tube and mix well. React in the dark at room temperature for 5 minutes. Pipette out 180 μL with a multi-channel pipette into a black 96-well plate and measure the fluorescence absorption value with a multimode microplate reader (INFINITE M1000). The excitation wavelength is 450 nm and the absorption wavelength is 482 nm. Using the fluorescence absorption value of the blank control as a reference, calculate the self-aggregation inhibition rate of the compound on Aβ 42 protein according to the following formula.

[0158] Inhibition rate = (1 - I Fi / I Fc ) × 100%, where I Fi is the fluorescence value of the sample containing the compound minus the background, and I Fc is the fluorescence absorption value of the blank control minus the background.

[0159] The results are shown in Table 1. The results indicate that when the ratio of the compound to Aβ is 1:2.5, most of the compounds described in the present invention have a certain inhibitory effect on Aβ 42 protein, among which Compound 3x exhibits the strongest inhibitory activity, reaching 69.0%, comparable to the positive controls curcumin and resveratrol. Therefore, the coumarin-8-hydroxyquinoline derivatives described in the present invention have great development prospects and can be used to prepare drugs for treating Alzheimer's disease.

[0160] Table 1 Inhibitory activity of coumarin-8-hydroxyquinazoline derivatives on Aβ 42 self-aggregation

[0161]

[0162] Example 36: Inhibitory effect of the quinoline derivatives described in the present invention on monoamine oxidase:

[0163] Select the compounds prepared in Examples 13 to 34, and use fluorescence method to determine the inhibitory activities of monoamine oxidase A and monoamine oxidase B. Clorgyline is used as the positive control for monoamine oxidase A, and Ladostigil is used as the positive control for monoamine oxidase B.

[0164] (1) Preparation of drug solutions:

[0165] Weigh a certain amount of each sample to be analyzed and dissolve it in dimethyl sulfoxide (DMSO), prepare a 10 mM concentration, store it in a -20 °C low-temperature refrigerator, and dilute it to the required concentration with phosphate buffer (50 mM / L, pH 7.4) when in use.

[0166] (2) Preparation of reaction solutions:

[0167] Monoamine oxidase A (M7316-1VL, recombinant, expressed in baculovirus infected BTI insect cells) was purchased from Sigma; pipette a certain volume of monoamine oxidase A and dilute it to 75 μg / mL with deionized water.

[0168] Monoamine oxidase B (M7441-1VL, recombinant, expressed in baculovirus infected BTI insect cells) was purchased from Sigma; pipette a certain volume of monoamine oxidase B and dilute it to 12.5 μg / mL with deionized water.

[0169] Horseradish peroxidase (SRE0082-5KU) was purchased from Sigma. A certain amount of Amplex Red reagent was weighed and prepared into a solution of 10 U / mL with phosphate buffer solution (50 mM / L, pH 7.4), and stored in the dark at 4 °C.

[0170] Amplex Red reagent was purchased from Sigma. A certain amount of Amplex Red reagent was weighed and prepared into a solution of 200 μM / L with phosphate buffer solution (50 mM / L, pH 7.4), and stored in the dark at 4 °C.

[0171] Tyramine was purchased from Sigma. A certain amount of Amplex Red reagent was weighed and prepared into a solution of 10 mM / L with phosphate buffer solution (50 mM / L, pH 7.4), and stored at 4 °C.

[0172] Benzylamine was purchased from Sigma. A certain amount of Amplex Red reagent was weighed and prepared into a solution of 10 mM / L with phosphate buffer solution (50 mM / L, pH 7.4), and stored at 4 °C.

[0173] (3) Monoamine oxidase test:

[0174] 80 μL of MAO-A or MAO-B enzyme solution and 20 μL of the test compound solution were respectively added into a black 96-well plate, incubated at 37 °C for 15 min, and immediately 20 μL of horseradish peroxidase solution, 20 μL of Amplex Red reagent solution and tyramine (for testing MAO-A) or benzylamine (for testing MAO-B), a total of 100 μL of a mixed solution of 40 μL of phosphate buffer solution were added. After incubation at 37 °C for 20 min, the fluorescence value was read at an excitation wavelength of 545 nm and an emission wavelength of 590 nm. The experimental results are shown in Table 2. The results show that some of the compounds described in the present invention have certain monoamine oxidase inhibitory activities. Among them, the inhibitory activities of 3a, 3g, 3h, 3n, 3q, 3t against MAO-B are significantly better than the positive control, and the selectivity index of 3n for oxidase B reaches 25.9. It can be used to prepare drugs for treating Alzheimer's disease.

[0175] Table 2 Inhibitory activities of coumarin-8-hydroxyquinazoline derivatives against monoamine oxidase

[0176]

[0177]

[0178] Inhibitory activity of monoamine oxidase at a compound concentration of 20 μM.

[0179] b Monoamine oxidase B selectivity index = IC 50 (hMAO-A) / IC50(hMAO-B).

[0180] c not determined.

[0181] Example 37: Inhibitory effect of the quinoline derivatives of the present invention on butyrylcholinesterase:

[0182] The inhibitory effects of the compounds prepared in Examples 13 to 34 on butyrylcholinesterase were tested using the method of Ellman (Biochemical Pharmacology 1961, 7, 88 - 95.). The results were expressed as IC 50 values, with Tacrine as the positive control. All tests were carried out on a PowerWave XS2 full-wavelength microplate reader at 37°C. Data analysis was processed using the software Origin.

[0183] (1) Preparation of drug solutions:

[0184] Weigh a certain amount of each sample to be analyzed and dissolve it in dimethyl sulfoxide (DMSO) to prepare a 10 mM concentration solution, which was stored in a -20°C refrigerator. Before use, it was diluted to the required concentration with phosphate buffer (0.1 mol / L, pH 8.0) to make the final DMSO concentration less than or equal to 0.5% (v / v).

[0185] (2) Preparation of enzyme stock solutions:

[0186] Butyrylcholinesterase (E.C. 3.1.1.8, from equine serum) was purchased from Sigma; weigh a certain amount of butyrylcholinesterase and dilute it with deionized water to the appropriate activity range.

[0187] (3) Preparation of substrate stock solutions:

[0188] Butylthiocholine (BTC) was purchased from Sigma; weigh a certain amount of BTC and prepare a 0.01 mol / L solution with phosphate buffer solution (0.1 mol / L, pH 8.0), and store it in the dark at 4°C.

[0189] (4) Preparation of chromogenic reagent stock solutions:

[0190] The chromogenic reagent 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) was purchased from Sigma. A certain amount of DTNB was weighed and prepared into a 0.01 mol / L solution with phosphate buffer solution (0.1 mol / L, pH 8.0), and stored in the dark at 4 °C.

[0191] (5) Testing:

[0192] Six wells were selected in a 96-well plate, and 10 μL of enzyme solution and 0, 5, 10, 20, 35, 50 μL of the compound solution to be tested were added respectively. Then 0.1 mol / L phosphate buffer solution with pH 8.0 was added to make the total volume 100 μL. It was incubated in a microplate reader with full wavelength at 37 °C for 15 min. Immediately, 100 μL of a mixture of 10 μL of BTC solution, 10 μL of DTNB solution and 80 μL of phosphate buffer solution was added, and the absorbance change was measured by scanning at λ = 412 nm for 2 min. The experimental results are shown in Table 3. The results indicate that some of the compounds described in the present invention have certain inhibitory activity against butyrylcholinesterase and can be used to prepare drugs for treating Alzheimer's disease.

[0193] Table 3. Inhibitory activity of compounds against butyrylcholinesterase

[0194]

[0195] a Inhibitory activity of butyrylcholinesterase at 5 μM of the compound.

[0196] Example 38: Blood-brain barrier permeability of the quinoline derivatives described in the present invention:

[0197] The compounds prepared in Examples 13 - 34 were selected and the blood-brain barrier permeability was predicted by the PAMPA-BBB method. 1. Reagent preparation

[0198] (1) Preparation of 2% porcine brain extract (Porcine brain lipid, PBL): Weigh 10 mg of porcine brain extract and add it to 500 μL of n-dodecane to dissolve it completely. Prepare it before use.

[0199] (2) Preparation of 50 mM PBS: Weigh 1.36 g of K2HPO4 into 200 mL of ultrapure water, and adjust the pH to 7.4 with KOH.

[0200] (3) Preparation of 5 mg / mL control drug stock solution: Weigh 5 mg of the control drug and dissolve it in 1 mL of DMSO, and store it at -20 °C. (4) Preparation of 100 μg / mL sample solution to be tested: Take 20 μL of the stock solution into a 1.5 mL EP tube, and add 980 μL of buffer solution (pH 7.4 PBS:ethanol = 70:30).

[0201] 2. Testing

[0202] (1) Carefully aspirate 4 μL of 2% porcine brain extract and add it onto the hydrophobic membrane of the 96-well plate serving as the dosing chamber.

[0203] (2) Quickly aspirate 200 μL of the sample solution to be tested into the 96-well plate as the dosing chamber, and add 300 μL of buffer solution (pH 7.4 PBS: ethanol: DMSO = 68:30:2) into the receiving chamber.

[0204] (3) Carefully place the dosing chamber flat on the receiving chamber, and ensure that the membrane is in full contact with the receiving liquid.

[0205] (4) Let it stand at room temperature for 10 hours, and then carefully remove the dosing chamber. Use a multifunctional microplate reader to measure the OD value of the compound in the receiving chamber at its maximum absorption peak.

[0206] (5) Aspirate 200 μL of the sample solution to be tested into 300 μL of buffer solution (pH 7.4 PBS: ethanol: DMSO = 68:30:2) and mix well to serve as the theoretical equilibrium solution, and measure the OD value of the compound at its maximum absorption peak.

[0207] (6) Use the buffer solution containing 2% DMSO as the blank control.

[0208] (7) Calculate the Pe value according to the formula

[0209] P e = -Vd×Va / [(Vd + Va)A×t] × ln(1 - OD 接收池内待测样品 / OD 理论平衡溶液待测样品 )

[0210] Vd is the volume of the dosing chamber, Va is the volume of the receiving chamber, A is the permeation area, and t is the permeation time.

[0211] The experimental results are shown in Table 4. The results indicate that the Pe values of most of the compounds described in the present invention are greater than 3.8, showing the ability to cross the blood-brain barrier. They can be used to prepare drugs for treating Alzheimer's disease.

[0212] Table 4 Prediction of Pe Values and Blood-Brain Barrier Permeation Ability of Coumarin-8-hydroxyquinazoline Derivatives

[0213] Compound <![CDATA[Pe(×10 -6 cm / s)]]> <![CDATA[prediction a > Compound <![CDATA[Pe(×10 -6 cm / s)]]> <![CDATA[prediction a > 3a 3.3±0.5 CNS± 3m 10.1±0.4 CNS+ 3b 14.2±1.5 CNS+ 3n 5.0±0.3 CNS+ 3c 14.5±1.1 CNS+ 3o 16.5±1.0 CNS+ 3d 0.8±0.1 CNS- 3p 3.8±1.0 CNS± 3e 4.5±0.4 CNS+ 3q 5.4±0.5 CNS+ 3f 5.5±0.0 CNS+ 3r 13.1±0.2 CNS+ 3g 7.5±0.2 CNS+ 3s 22.2±1.8 CNS+ 3h 1.3±0.0 CNS- 3t 7.7±0.5 CNS+ 3i 14.0±0.8 CNS+ 3u 19.7±1.5 CNS+ 3j 4.8±0.5 CNS+ 3v 10.6±0.4 CNS+ 3k 13.6±1.3 CNS+ 3w 5.7±0.5 CNS+ 3l 13.6±0.8 CNS+ 3x 12.4±0.7 CNS+

[0214] aCompounds with permeabilities Pe>3.8×10-6cm / s could cross the BBB by passive diffusion, CNS+, high brain penetration;CNS-, low brain penetration n.d., no determined.

[0215] Example 39: Protective effect of quinoline derivative 3x of the present invention on Aβ-induced cytotoxicity in SYSH-5Y cells

[0216] Select the compound prepared in Example 34, and use the MTT method to evaluate the protective effect of the compound on Aβ-induced cytotoxicity in SYSH-5Y cells.

[0217] 1. Incubation of the drug

[0218] (1) Dissolve Aβ42 in DMSO to 5 mM, and then dilute it with DMEM medium to 20 μM for standby.

[0219] (2) Take an appropriate volume of 20 μM Aβ 42， Add different volumes of 10 mM 3x to make the final concentrations 2, 5, and 10 μM respectively, and incubate at 37 °C for 24 hours.

[0220] 2. Operation process

[0221] Take SYSH-5Y cells in the logarithmic growth phase, digest them with 0.25% trypsin, centrifuge to collect the cells, resuspend them with DMEM medium (10% fetal bovine serum, 1% double antibody), count them with a cell counting plate, and inoculate them into a 96-well cell culture plate, 100 μL per well, 5000 cells / well. After culturing for 24 hours to allow the cells to adhere, aspirate the original medium, add the incubated Aβ42 and 3x, 100 μL per well, and set 3 replicates. Continue to culture for 24 hours. The blank and control groups are added with medium containing the same amount of DMSO instead. After 24 hours, except for the blank group wells, add medium containing 5 mg / mL MTT to each well and continue to culture for 4 hours. Then discard the medium, and add 100 μL DMSO to each well. After the product is fully dissolved, measure the OD value of each well at a wavelength of 570 nm on a full-wavelength microplate reader. The cell survival rate in each sample is calculated by the following formula: Cell survival rate (%) = (ODsample - ODblank) / (ODcontrol - ODblank). The results are as Figure 1 shown. The results show that the compound 3x of the present invention has a strong protective ability against Aβ42-induced damage to SYSH-5Y cells and can be used to prepare drugs for treating Alzheimer's disease.

[0222] Example 40: Effect of the quinoline derivative 3x of the present invention on the cognitive ability of APP / PS1 transgenic mice

[0223] The compound prepared in Example 34 was selected, and the water maze method was used to evaluate the effect of the compound on the cognitive ability of APP / PS1 transgenic mice

[0224] 1. Animals

[0225] Eighteen SPF-grade 6-month-old APPsw / PSEN1 double transgenic male mice and six wild-type mice C57BL / 6J of the same species were purchased from Huafukang Biotechnology Co., Ltd. The newly purchased mice were caged and fed indoors with the room temperature controlled at about 23°C, 12-hour light, humidity 60 - 70%, free access to food and water, and acclimated to the environment in the experimental animal center for one week. All experiments complied with the guidelines for the use and care of experimental animals and were approved by the Animal Experiment Ethics Committee of Zunyi Medical University. The experimental animal use license number is SYXK(Qian)2021 - 0004

[0226] 2. Grouping

[0227] Six wild-type mice C57BL / 6J of the same species were used as the wild-type control group. Eighteen APPsw / PSEN1 double transgenic mice were randomly divided into three groups of 6 each, namely the normal saline group, the positive control donepezil group, and the 3x treatment group

[0228] 3. Water maze place navigation experiment

[0229] The water maze place navigation test was performed 24 days after gavage. First, clear water about 32 cm deep was injected into the pool (diameter 120 cm, depth 45 cm) so that the water surface was 2 cm higher than the platform (diameter 9 cm, height 30 cm). The water temperature was controlled at about 20°C, and titanium dioxide was added to make the water milky white so that the safety platform could not be clearly seen in the water. The pool was equally divided into 4 quadrants (from east, south, west, and north are quadrants one to four respectively). The platform was placed in the middle of the third quadrant, and the mice were placed into the water at any point in the four quadrants facing the pool wall. The test lasted for 5 days. Four training sessions were carried out every day, and the mice entered the water from 4 different entry points, 90 seconds each time. The instrument automatically recorded the route map of the mice finding and climbing onto the platform, the time (latency) required for the mice to find the underwater hidden platform and stand on it from entering the water, and the swimming distance. If the mice did not find the platform within 90 seconds, they needed to be guided to the platform, and the latency was counted as 90 seconds at this time. Let the mice stay for 10 seconds and then put them back into the cage

[0230] 4. Water maze spatial exploration experiment

[0231] Twenty-four hours after the positioning navigation test was completed, the platform was removed. The mice entered the water in the quadrant opposite to the original platform quadrant, and the number of times the mice crossed the platform within 90 seconds was recorded and compared and analyzed. Data collection and processing were completed by Morris software. The results are as Figure 2 shown. The results show that compound 3x of the present invention can significantly shorten the time for AD mice to find the platform and increase the number of times of passing through the virtual platform, improve the spatial cognitive ability of AD mice, and can be used to prepare drugs for treating Alzheimer's disease.

Claims

1. Coumarin-quinoline derivatives, characterized in that: The compound has the structure shown in formula (I) or formula (II), , , the structure represented by the formula (I) or formula (II) is any one of the following compounds: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 2. The preparation method of the coumarin-quinoline derivative according to claim 1, characterized in that: Coumarin structures with anti-Aβ 42 protein aggregation activity are fused with the pharmacophore quinoline ring of anti-Alzheimer's disease drugs, while changing the length between the two structures.

3. The preparation method of the coumarin-quinoline derivative according to claim 2, characterized in that: The preparation process of the said formula (I) is as follows: (1) Dissolve and bromohydrocarbon alkane in solvent A, and carry out a substitution reaction under the action of acid-binding agent B to obtain compound ; the solvent A is one or a mixture of several reagents selected from acetone, dichloromethane, chloroform, acetonitrile, tetrahydrofuran, dioxane or dimethylformamide; (2) React with 5-chloro-8-hydroxyquinoline or 8-hydroxyquinoline to obtain the target product .

4. The preparation method of the coumarin-quinoline derivative according to claim 2, wherein: The preparation process of the said formula (II) is as follows: (1) Dissolve and bromoalkane in solvent A, and carry out a substitution reaction under the action of acid-binding agent B to obtain compound ; the solvent A is one or a mixture of several reagents selected from acetone, dichloromethane, chloroform, acetonitrile, tetrahydrofuran, dioxane or dimethylformamide; (2) React with 5-chloro-8-hydroxyquinoline or 8-hydroxyquinoline to obtain the target product .

5. The preparation method of the coumarin-quinoline derivative according to claim 3 or 4, characterized in that: The said brominated alkane is 1,2-dibromoethane, 1,3-dibromopropane, 1,4-dibromobutane or 1,5-dibromopentane.

6. The preparation method of the coumarin-quinoline derivative according to claim 3 or 4, characterized in that: The used acid-binding agent B is one or a mixture of reagents of potassium carbonate, sodium carbonate, cesium carbonate, triethylamine, N,N-diisopropylethylamine.

7. Use of the coumarin-quinoline derivative according to claim 1 in the preparation of a medicament for treating Alzheimer's disease.

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