Substituted coumarin-eugenol derivative as well as preparation method and application thereof

Abstract

The invention discloses a substituted coumarin-eugenol derivative as well as a preparation method and application thereof. The derivative is prepared by connecting a coumarin mother nucleus and eugenol through a flexible chain by virtue of a two-step nucleophilic substitution reaction. The derivative disclosed by the invention is novel in structure, can simultaneously and effectively inhibit monoamine oxidase-B and acetylcholin esterase and inhibit A beta protein aggregation, has excellent oxidation resistance and metal ion chelation capability, and realizes a multi-target synergistic effect. In-vitro and in-vivo experiments show that the compound has a remarkable improvement effect on a plurality of pathological links such as Alzheimer's disease, and is low in neurotoxicity and high in safety. Meanwhile, the preparation method has the advantages of easily available raw materials and simple steps, is suitable for industrial production, and has a wide application prospect in the aspect of preparing the anti-neurodegenerative disease medicine.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to a substituted coumarin-eugenol derivative, a preparation method and application thereof. BACKGROUND

[0002] Alzheimer's disease (AD), also known as senile dementia, is a chronic progressive central nervous system degenerative disease. With the development of society and the improvement of people's living standards, the world population is becoming increasingly aged, and therefore the incidence of AD is increasing year by year, and has become one of the major diseases that seriously threaten the life and health and quality of life of the elderly. So far, the pathogenesis of AD has not been clear, and many factors are involved, such as the misfolding and aggregation of β-amyloid protein, the decrease of cholinergic level, inflammation, oxidative stress, the over-phosphorylation of tau protein, mitochondrial damage, abnormal energy metabolism, imbalance of metal ions in the body, and abnormal cell apoptosis cycle. These factors are interrelated and influence each other, and therefore the therapeutic effect of single target is mostly unsatisfactory.

[0003] Among the existing AD treatment drugs, most of them still focus on single target, for example, acetylcholinesterase inhibitors (such as donepezil) can only relieve symptoms and cannot stop disease progression; and the immunotherapy targeting Aβ has been repeatedly frustrated in clinical trials, reflecting the limitations of single intervention strategy under the complex pathological mechanism of AD. Therefore, in recent years, multi-target drugs have become one of the hotspots in the research of anti-AD drugs. A large number of experiments have proved that the activity of MAO-B gradually increases with age, especially in the elderly around the AD plaque, and the increase of its activity will lead to an increase in free radicals with neurotoxicity in the brain, increase of oxidative stress, and further accelerate the aggregation of Aβ protein and the over-phosphorylation of tau protein in the brain, and finally cause nerve damage and lead to neuronal death. Inhibitors of MAO-B have also been proved to have important value in the treatment of AD (Zhang N. Research Progress of Monoamine Oxidase Inhibitors in the Treatment of Neurodegenerative Diseases [J]. Medical Theory and Practice, 2015, 28(13): 1713-1715, 1718).

[0004] On the other hand, oxidative stress also plays a very important role in the pathogenesis of Alzheimer's disease (AD). Abnormal metabolism of substances and energy in the brain tissue of AD patients leads to the accumulation of free radicals, resulting in oxidative damage, which in turn causes apoptosis and characteristic neuropathological changes in AD. Numerous experiments have demonstrated that antioxidants have a protective effect against AD and can slow down its progression (Chen Mengyuan, Zhang Juan, Li Chao, et al. Potential preventive and therapeutic effects of nitric oxide and natural antioxidants on Alzheimer's disease [J]. Food and Nutrition Science, 2016, 005(003): 105-113). Therefore, research on small molecules with antioxidant effects has become one of the hot topics in anti-AD drug research.

[0005] Furthermore, imbalance of metal ion homeostasis is also considered an important link in the pathogenesis of Alzheimer's disease (AD). Metal ions such as copper, zinc, and iron can promote Aβ aggregation and increase its neurotoxicity, while metal ion chelators are considered to have potential therapeutic value. Although acetylcholinesterase inhibition is a well-established target, existing drugs have drawbacks such as significant side effects and limited efficacy. Therefore, developing drugs that can simultaneously target multiple key targets, including MAO-B, Aβ aggregation, oxidative stress, metal ion imbalance, and the cholinergic system, has become an important direction in the current research and development of new anti-AD drugs.

[0006] Coumarins and eugenol are both bioactive structural units found in natural products. Coumarin compounds possess various pharmacological activities, including anti-inflammatory, antioxidant, and anti-Aβ aggregation effects, while eugenol exhibits excellent antioxidant and neuroprotective properties. However, there are currently no reports on the systematic study of multi-target anti-AD activity by linking coumarins and eugenol through flexible chains. Therefore, designing and synthesizing a class of novel substituted coumarin-eugenol derivatives and evaluating their activity at multiple AD-related targets is of great significance for the development of novel multi-target anti-AD drugs. Summary of the Invention

[0007] To address the aforementioned technical problems, this invention proposes a substituted coumarin-eugenol derivative, its preparation method, and its application, in order to overcome the shortcomings and deficiencies of existing anti-AD drugs, which have single targets and limited therapeutic effects. This compound can simultaneously inhibit MAO-B and Aβ self-aggregation, and also has antioxidant properties.

[0008] To achieve the above objectives, the present invention provides a substituted coumarin-eugenol derivative having a structure as shown in general formula (I) or general formula (II):

[0009] Wherein, R1 is selected from hydrogen, methyl, chloro or formyloxyethyl; R2 is selected from hydrogen, methyl, trifluoromethyl or phenyl; n is an integer from 2 to 5.

[0010] Furthermore, the derivative is any one of the following compounds: .

[0011] The present invention also provides pharmaceutically acceptable salts of the substituted coumarin-eugenol derivatives as described above.

[0012] The present invention also provides a method for preparing the substituted coumarin-eugenol derivative as described above, comprising the following steps: S1. React the first or second reactant with Br-(CH2). n -Br undergoes a monosubstitution reaction in an alkaline environment to give either the first or second intermediate product; S2. The first or second intermediate obtained in step S1 is reacted with eugenol under alkaline conditions to undergo a nucleophilic substitution reaction to obtain the substituted coumarin-eugenol derivative of general formula (I) or general formula (II). The structure of the first reactant is as follows: The structure of the second reactant is In this context, R1 is selected from hydrogen, methyl, chloro, or formyloxyethyl; R2 is selected from hydrogen, methyl, trifluoromethyl, or phenyl; and n is an integer from 2 to 5.

[0013] Further, in step S1, the alkaline conditions are provided by cesium carbonate, potassium carbonate, or sodium carbonate; and / or, in step S2, the alkaline conditions are provided by cesium carbonate or potassium carbonate; and / or, the solvent in step S2 is N,N-dimethylformamide or acetonitrile.

[0014] Further, in step S1, the monosubstituted reaction is carried out at 25–60 °C for 2–8 h; and / or, in step S2, the nucleophilic substitution reaction is carried out at 25–100 °C for 2–8 h.

[0015] The present invention also provides the use of the substituted coumarin-eugenol derivatives as described above or pharmaceutically acceptable salts thereof in the preparation of medicaments for inhibiting monoamine oxidase-B, inhibiting acetylcholinesterase, inhibiting Aβ protein aggregation, antioxidation, and / or chelating metal ions.

[0016] The present invention also provides the use of the substituted coumarin-eugenol derivatives as described above or pharmaceutically acceptable salts thereof in the preparation of medicaments for the prevention and / or treatment of neurodegenerative diseases.

[0017] Furthermore, the neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, vascular dementia, Huntington's disease, amyotrophic lateral sclerosis (ALS), or myasthenia gravis.

[0018] The present invention also provides a pharmaceutical formulation comprising a therapeutically effective amount of the substituted coumarin-eugenol derivative as described above or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients.

[0019] Furthermore, the dosage form of the pharmaceutical preparation is tablets, pills, capsules, injections, suspensions, or emulsions.

[0020] Compared with the prior art, the present invention has the following advantages and technical effects: This invention provides a substituted coumarin-eugenol derivative with a novel structure. This compound exhibits inhibitory effects on monoamine oxidase-B activity, Aβ aggregation activity, and antioxidant effects. All of these targets are closely related to diseases such as Alzheimer's disease. Through the combined action of multiple targets, it can effectively reverse scopolamine-induced memory impairment in mice, demonstrating that this compound has significant therapeutic effects on diseases such as Alzheimer's disease. Furthermore, this substituted coumarin-eugenol derivative has low biotoxicity and high safety, making it highly valuable for medical research and market applications. On the other hand, its production requires inexpensive raw materials, involves few reaction steps, and is simple to prepare, making it highly suitable for large-scale industrial production. Attached Figure Description

[0021] Figure 1 The UV spectrum of the interaction between the substituted coumarin-eugenol derivative Dg-4d and metal ions in Experimental Example 11; Figure 2 The cell viability graphs are obtained from Examples 11 and 12 of Experiment 7, showing the cell viability of H2O2-induced SH-SY5Y cells after treatment with the substituted coumarin-eugenol derivatives Dg-4d and Dg-5d obtained in Examples 11 and 12. Figure 3 The substituted coumarin-eugenol derivative Dg-4d in Example 11 is an example of the effect of Dg-4d on Aβ. 1-42 Figure 1 shows the results of a passive avoidance experiment that induced the improvement of memory impairment in mice; where A shows the change in the latency of mice entering the dark room, and B shows the number of errors during the 5-minute test period. Detailed Implementation

[0022] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0023] This invention provides a series of novel substituted coumarin-eugenol derivatives, whose general preparation method mainly involves a two-step substitution reaction. The synthetic route is smooth and simple to operate, as shown below:

[0024] The general preparation steps are briefly described below: S1. The coumarin nucleus compound shown in formula (III) or (V) is reacted with an excess of 1,ω-dibromoalkane (Br-(CH2)). n -Br, n=2~5) undergoes a monosubstitution reaction in an alkaline environment to give intermediate compound (Ⅳ) or (Ⅵ); S2. The intermediate obtained in step S1 is subjected to a nucleophilic substitution reaction with eugenol under alkaline conditions. After post-treatment purification, the target product compound (I) or (II) is obtained.

[0025] The synthesis process of representative compounds is described in detail below through specific examples.

[0026] Example 1 Synthesis of Dg-2b, a substituted coumarin-eugenol derivative

[0027] In a 25 mL round-bottom flask, add 5 mL of acetonitrile (CH3CN), eugenol (0.21 g, 1.23 mmol), and anhydrous cesium carbonate (0.28 g, 1.99 mmol). After stirring the mixture at room temperature for 30 min, add the coumarin intermediate 7-(2-bromoethoxy)-4-methyl-2 H -Crotene-2-one (0.2 g, 1.30 mmol) was stirred at 80 °C, and the reaction progress was monitored by TLC. After the reaction was completed, the reaction solution was evaporated under reduced pressure, water was added and stirred at room temperature for 2 h to allow the solid to fully precipitate. The solid was then filtered, dried, and separated by silica gel column chromatography to obtain the coumarin-eugenol derivative Dg-2b, with a yield of 29%.

[0028] 1 H NMR (600 MHz, CDCl3) δ 7.49 (d, J = 8.7 Hz, 1H), 6.93 - 6.88 (m,3H), 6.73 (d, J = 8.5 Hz, 2H), 6.14 (d, J = 1.7 Hz, 1H), 5.96 (ddt, J = 16.8,10.1, 6.7 Hz, 1H), 5.14 - 5.02 (m, 2H), 4.39 (s, 4H), 3.85 (s, 3H), 3.34 (d, J = 6.6 Hz, 2H), 2.40 (d, J = 1.3 Hz, 3H). 13C NMR (151 MHz, CDCl3) δ 161.77,161.29, 155.22, 152.51, 149.86, 146.21, 137.52, 134.22, 125.52, 120.55,115.78, 114.89, 112.77, 112.61, 112.14, 101.78, 67.93, 67.19, 55.90, 39.85,18.69. ESI-MS m / z: 366.15 [M+H] + .

[0029] Example 2 Synthesis of Dg-3b, a substituted coumarin-eugenol derivative

[0030] The difference from Example 1 is that the raw material used in this example is 7-(3-bromopropoxy)-4-methyl-2 H -Creno-2-one replaced by 7-(2-bromoethoxy)-4-methyl-2 H -Crotene-2-one, other parameters and operations are as described in Example 1, to obtain the substituted coumarin-eugenol derivative Dg-3b, yield: 39%.

[0031] 1 H NMR (600 MHz, CDCl3) δ 7.47 (d, J = 8.7 Hz, 1H), 6.88 - 6.82 (m,3H), 6.75 - 6.66 (m, 2H), 6.12 (t, J = 1.1 Hz, 1H), 5.95 (ddt, J = 16.8,10.1, 6.7 Hz, 1H), 5.16 - 4.99 (m, 2H), 4.22 (dt, J = 28.5, 6.1 Hz, 4H), 3.84(s, 3H), 3.32 (d, J = 6.7 Hz, 2H), 2.39 (t, J = 1.0 Hz, 3H), 2.32 (t, J = 6.1Hz, 2H). 13C NMR (151 MHz, CDCl3) δ 162.02, 161.34, 155.29, 152.54, 149.55,146.53, 137.62, 133.38, 125.49, 120.48, 115.67, 113.74, 113.58, 112.60,112.42, 111.96, 101.54, 65.58, 65.16, 55.90, 39.82, 29.12, 18.68. ESI-MS m / z:381.17 [M+H] + .

[0032] Example 3 Synthesis of Dg-4b, a substituted coumarin-eugenol derivative

[0033] The difference from Example 1 is that the raw material used in this example is 7-(4-bromobutoxy)-4-methyl-2 H -Creno-2-one replaced by 7-(2-bromoethoxy)-4-methyl-2 H -Crotene-2-one, other parameters and operations are as described in Example 1, to obtain the substituted coumarin-eugenol derivative Dg-4b, yield: 28%.

[0034] 1 H NMR (600 MHz, CDCl3) δ 7.47 (d, J = 8.8 Hz, 1H), 6.85 - 6.79 (m,3H), 6.70 (d, J = 9.9 Hz, 2H), 6.11 (s, 1H), 5.95 (ddt, J = 16.9, 10.3, 6.8Hz, 1H), 5.11 - 5.02 (m, 2H), 4.08 (dd, J = 22.7, 5.5 Hz, 4H), 3.84 (s, 3H), 3.33 (d, J = 6.7 Hz, 2H), 2.38 (s, 3H), 2.02 (q, J = 3.8 Hz, 4H). 13C NMR (151MHz, CDCl3) δ 162.14, 161.35, 155.30, 152.59, 149.43, 146.66, 137.67, 133.04,125.48, 120.45, 115.65, 113.47, 113.30, 112.66, 112.35, 111.86, 101.36,68.69, 68.24, 55.91, 39.83, 26.00, 25.87, 18.68. ESI-MS m / z: 395.19 [M+H] + .

[0035] Example 4 Synthesis of Dg-5b, a substituted coumarin-eugenol derivative

[0036] The difference from Example 1 is that the raw material used in this example is 7-(5-bromopentoxy)-4-methyl-2 H -Creno-2-one replaced by 7-(2-bromoethoxy)-4-methyl-2 H -Crotene-2-one, other parameters and operations are as described in Example 1, to obtain the substituted coumarin-eugenol derivative Dg-5b, yield: 28%.

[0037] 1 H NMR (600 MHz, CDCl3) δ 7.48 (dd, J = 8.9, 2.0 Hz, 1H), 6.86 - 6.79(m, 3H), 6.70 (d, J = 9.6 Hz, 2H), 6.13 (d, J = 2.5 Hz, 1H), 6.00 - 5.91 (m,1H), 5.11 - 5.02 (m, 2H), 4.04 (q, J = 7.6 Hz, 4H), 3.85 (d, J = 1.9 Hz, 3H), 3.33 (d, J = 6.6 Hz, 2H), 2.39 (s, 3H), 1.90 (q, J = 7.6 Hz, 4H), 1.68 (d, J = 7.0 Hz, 2H). 13C NMR (151 MHz, CDCl3) δ 162.17, 161.38, 155.32, 152.57,149.43, 146.78, 137.69, 132.93, 125.48, 120.46, 115.62, 113.48, 113.32,112.63, 112.38, 111.88, 101.40, 68.93, 68.38, 55.95, 39.82, 28.98, 28.79,22.65, 18.69.ESI-MS m / z: 409.20 [M+H] + .

[0038] Example 5 Synthesis of Dg-2c, a substituted coumarin-eugenol derivative

[0039] In a 25 mL round-bottom flask, add 5 mL of N,N-dimethylformamide (DMF), eugenol (0.21 g, 1.23 mmol), and anhydrous potassium carbonate (0.30 g, 2.16 mmol). After stirring the mixture at room temperature for 30 min, add the coumarin intermediate 4-(2-bromoethoxy)-2 H -Crotene-2-one (0.29 g, 1.08 mmol) was stirred at 80 °C, and the reaction progress was monitored by TLC. After the reaction was completed, water was added to the reaction solution and stirred at room temperature for 2 h to allow the solid to precipitate fully. The solid was then filtered, dried, and separated by silica gel column chromatography to obtain the coumarin-eugenol derivative Dg-2c, with a yield of 61%.

[0040] 1 H NMR (600 MHz, CDCl3) δ 7.82 (dd, J = 8.0, 1.7 Hz, 1H), 7.57 (ddd, J = 8.5, 7.3, 1.6 Hz, 1H), 7.35 - 7.25 (m, 2H), 6.95 (d, J = 8.2 Hz, 1H), 6.77(d, J = 6.8 Hz, 2H), 5.99 (ddt, J = 16.8, 9.8, 6.7 Hz, 1H), 5.77 (s, 1H), 5.16 - 5.09 (m, 2H), 4.51 (s, 4H), 3.87 (s, 3H), 3.38 (d, J= 6.7 Hz, 2H). 13 CNMR (151 MHz, CDCl3) δ 165.55, 162.92, 153.35, 149.63, 146.33, 137.55,133.70, 132.37, 123.85, 122.98, 120.45, 116.80, 115.74, 115.72, 113.94,112.40, 90.68, 66.11, 65.44, 55.84, 39.83, 28.72. ESI-MS m / z: 353.14 [M+H] + .

[0041] Example 6 Synthesis of Dg-3c, a substituted coumarin-eugenol derivative

[0042] In a 25 mL round-bottom flask, add 5 mL of DMF, eugenol (0.32 g, 1.95 mmol), and anhydrous cesium carbonate (0.99 g, 3.25 mmol). After stirring the mixture at room temperature for 30 min, add the coumarin intermediate 4-(3-bromopropoxy)-2. H -Crotene-2-one (0.31 g, 1.08 mmol) was stirred at room temperature, and the reaction progress was monitored by TLC. After the reaction was completed, water was added to the reaction solution and stirred at room temperature for 2 h to allow the solid to precipitate fully. The solid was then filtered and dried. The coumarin-eugenol derivative Dg-3c was obtained by silica gel column chromatography with a yield of 52%.

[0043] 1 H NMR (600 MHz, CDCl3) δ 7.75 (dd, J = 7.9, 1.6 Hz, 1H), 7.49 (ddd, J = 8.7, 7.3, 1.6 Hz, 1H), 7.28 - 7.20 (m, 2H), 6.80 (d, J = 8.0 Hz, 1H), 6.68- 6.62 (m, 2H), 5.90 (ddt, J = 16.8, 10.0, 6.7 Hz, 1H), 5.68 (s, 1H), 5.07 -4.98 (m, 2H), 4.33 (t, J = 6.1 Hz, 2H), 4.19 (t, J= 6.0 Hz, 2H), 3.79 (s,3H), 3.28 (d, J = 6.7 Hz, 2H), 2.37 (p, J = 6.0 Hz, 2H). 13 C NMR (151 MHz, CDCl3) δ 165.55, 162.92, 153.35, 149.63, 146.33, 137.55, 133.70, 132.37,123.85, 122.98, 120.45, 116.80, 115.74, 115.72, 113.94, 112.40, 90.68, 66.11,65.44, 55.84, 39.83, 28.72. ESI-MS m / z: 367.15 [M+H] + .

[0044] Example 7 Synthesis of Dg-4c, a substituted coumarin-eugenol derivative

[0045] The difference from Example 6 is that the coumarin intermediate in this example is 4-(4-bromobutoxy)-2 H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-4c, yield: 81%.

[0046] 1 H NMR (600 MHz, CDCl3) δ 7.75 (dd, J = 7.9, 1.7 Hz, 1H), 7.52 - 7.47(m, 1H), 7.28 - 7.19 (m, 2H), 6.77 (d, J = 8.0 Hz, 1H), 6.69 - 6.64 (m, 2H), 5.91 (ddt, J = 16.7, 9.8, 6.8 Hz, 1H), 5.66 (s, 1H), 5.07 - 5.00 (m, 2H), 4.21 (t, J = 6.3 Hz, 2H), 4.07 (t, J = 6.1 Hz, 2H), 3.81 (s, 3H), 3.29 (d, J= 6.7 Hz, 2H), 2.10 (p, J = 6.7 Hz, 2H), 2.05 - 2.00 (m, 2H). 13 C NMR (151MHz, CDCl3) δ 165.67, 163.00, 153.35, 149.42, 146.47, 137.63, 133.18, 132.31,123.82, 123.04, 120.40, 116.75, 115.78, 115.68, 113.25, 112.28, 90.49, 69.20,68.55, 55.86, 39.82, 29.71, 25.77 . ESI-MS m / z: 381.17 [M+H] + .

[0047] Example 8 Synthesis of Dg-5c, a substituted coumarin-eugenol derivative

[0048] The difference from Example 6 is that the coumarin intermediate in this example is 4-(5-bromopentoxy)-2 H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-5c, yield: 68%.

[0049] 1 H NMR (600 MHz, CDCl3) δ 7.77 (dd, J = 7.8, 1.6 Hz, 1H), 7.49 (td, J = 7.8, 1.6 Hz, 1H), 7.26 - 7.19 (m, 2H), 6.76 (d, J = 7.9 Hz, 1H), 6.65 (d, J = 10.5 Hz, 2H), 5.95 - 5.87 (m, 1H), 5.62 (s, 1H), 5.05 - 4.99 (m, 2H), 4.11(t, J = 6.3 Hz, 2H), 4.00 (t, J = 6.4 Hz, 2H), 3.28 (d, J = 6.7 Hz, 2H), 1.97- 1.92 (m, 2H), 1.88 (q,J = 7.1 Hz, 2H), 1.68 (q, J = 8.2 Hz, 2H). 13 C NMR(151 MHz, CDCl3) δ 165.69, 163.01, 153.38, 149.44, 146.71, 137.66, 133.05,132.34, 123.85, 123.04, 120.46, 116.80, 115.81, 115.65, 113.36, 112.39,90.44, 69.22, 68.79, 55.92, 39.82, 28.90. ESI-MS m / z: 395.19 [M+H] + .

[0050] Example 9 Synthesis of Dg-2d, a substituted coumarin-eugenol derivative

[0051] The difference from Example 6 is that the coumarin intermediate in this example is 7-(2-bromoethoxy)-3,4-dimethyl-2 H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the coumarin-eugenol derivative Dg-2d, yield: 39%.

[0052] 1 H NMR (600 MHz, CDCl3) δ 7.49 (dd, J = 9.0, 2.0 Hz, 1H), 6.92 - 6.85(m, 3H), 6.76 - 6.69 (m, 2H), 5.95 (dtt, J = 14.7, 7.1, 3.5 Hz, 1H), 5.15 -5.02 (m, 2H), 4.40 - 4.36 (m, 4H), 3.87 - 3.83 (m, 3H), 3.34 (d, J = 6.7 Hz, 2H), 2.36 (d, J = 2.1 Hz, 3H), 2.18 (d, J = 2.3 Hz, 3H). 13C NMR (151 MHz, CDCl3) δ 162.44, 160.64, 153.48, 149.83, 146.25, 146.23, 137.54, 134.15,125.26, 120.55, 119.10, 115.76, 114.84, 114.44, 112.61, 112.54, 101.48,67.94, 67.09, 55.90, 39.84, 15.09, 13.17. ESI-MS m / z: 381.17 [M+H] + .

[0053] Example 10 Synthesis of Dg-3d, a substituted coumarin-eugenol derivative

[0054] The difference from Example 6 is that the coumarin intermediate in this example is 7-(3-bromopropoxy)-3,4-dimethyl-2 H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-3d, yield: 54%.

[0055] 1 H NMR (600 MHz, CDCl3) δ 7.47 (d, J = 8.8 Hz, 1H), 6.87 - 6.80 (m,3H), 6.73 - 6.68 (m, 2H), 5.99 - 5.87 (m, 1H), 5.10 - 5.00 (m, 2H), 4.22 (t, J = 6.1 Hz, 2H), 4.19 (t, J = 6.1 Hz, 2H), 3.84 (d, J = 1.4 Hz, 3H), 3.32(dd, J = 6.5, 1.9 Hz, 2H), 2.35 (s, 3H), 2.31 (p, J = 6.1 Hz, 2H), 2.17 (s, 3H). 13C NMR (151 MHz, CDCl3) δ 162.48, 160.89, 153.55, 149.54, 146.56,146.25, 137.63, 133.33, 125.21, 120.48, 118.91, 115.66, 114.19, 113.73,112.42, 112.38, 101.25, 65.63, 65.05, 55.91, 39.82, 29.15, 15.07, 13.16.ESI-MS m / z: 395.19 [M+H] + .

[0056] Example 11 Synthesis of Dg-4d, a substituted coumarin-eugenol derivative

[0057] The difference from Example 6 is that the coumarin intermediate in this example is 7-(4-bromobutoxy)-3,4-dimethyl-2 H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-4d, yield: 63%.

[0058] 1 H NMR (600 MHz, CDCl3) δ 7.47 (d, J = 8.8 Hz, 1H), 6.85 - 6.77 (m,3H), 6.70 (d, J = 8.4 Hz, 2H), 5.95 (dddd, J = 17.9, 10.0, 7.3, 6.2 Hz, 1H),5.11 - 5.03 (m, 2H), 4.09 (dd, J = 12.7, 6.2 Hz, 4H), 3.85 (d, J = 1.4 Hz, 3H), 3.33 (d, J = 6.7 Hz, 2H), 2.35 (d, J = 1.4 Hz, 3H), 2.17 (d, J = 1.6 Hz, 3H), 2.02 (p, J = 3.3 Hz, 4H). 13C NMR (151 MHz, CDCl3) δ 162.48, 161.01,153.57, 149.44, 146.68, 146.28, 137.68, 133.02, 125.20, 120.45, 118.82,115.64, 114.08, 113.32, 112.43, 112.36, 101.09, 68.70, 68.11, 55.92, 39.83,25.99, 25.90, 15.07, 13.16. ESI-MS m / z: 395.19 [M+H] + .

[0059] Example 12 Synthesis of Dg-5d, a substituted coumarin-eugenol derivative

[0060] The difference from Example 6 is that the coumarin intermediate in this example is 7-(5-bromopentoxy)-3,4-dimethyl-2 H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-5d, yield: 64%.

[0061] 1 H NMR (600 MHz, CDCl3) δ 7.48 (dd, J = 8.9, 1.8 Hz, 1H), 6.82 (ddd, J = 10.3, 7.4, 2.0 Hz, 2H), 6.78 (d, J = 2.2 Hz, 1H), 6.71 (dd, J = 9.3, 2.2Hz, 2H), 5.95 (dtt, J = 16.9, 8.9, 4.4 Hz, 1H), 5.13 - 5.02 (m, 2H), 4.06 -4.01 (m, 4H), 3.85 (d, J = 1.9 Hz, 3H), 3.33 (d, J = 6.7 Hz, 2H), 2.36 (s,3H), 2.18 (s, 3H), 1.94 - 1.86 (m, 4H), 1.69 - 1.66 (m, 2H). 13C NMR (151 MHz, CDCl3) δ 162.51, 161.04, 153.57, 149.42, 146.79, 146.30, 137.70, 132.91,125.21, 120.46, 118.82, 115.61, 114.08, 113.32, 112.41, 112.39, 101.10,68.94, 68.26, 55.95, 39.82, 28.99, 28.83, 22.65, 15.07, 13.15. ESI-MS m / z:423.22 [M+H] + .

[0062] Example 13 Synthesis of Dg-2e, a substituted coumarin-eugenol derivative

[0063] The difference from Example 6 is that the coumarin intermediate in this example is 7-(2-bromoethoxy)-4-phenyl-2- H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-2e, yield: 34%.

[0064] 1 H NMR (600 MHz, CDCl3) δ 7.51 (dd, J = 5.4, 2.7 Hz, 3H), 7.46 - 7.42(m, 2H), 7.38 (dd, J = 8.9, 2.1 Hz, 1H), 6.97 (q, J = 3.6 Hz, 1H), 6.90 (dt, J = 7.9, 3.9 Hz, 1H), 6.84 (dt, J = 8.8, 2.4 Hz, 1H), 6.75 - 6.71 (m, 2H), 6.25 - 6.20 (m, 1H), 5.96 (dddd, J = 16.8, 14.6, 6.6, 3.2 Hz, 1H), 5.12 -5.05 (m, 2H), 4.40 (dt, J = 5.5, 3.0 Hz, 4H), 3.85 (t, J = 3.8 Hz, 3H), 3.34(d,J = 6.7 Hz, 2H). 13 C NMR (151 MHz, CDCl3) δ 161.93, 161.25, 155.93,155.81, 149.85, 146.19, 137.52, 135.58, 134.22, 129.61, 128.85(2C), 128.41(2C), 127.99, 120.56, 115.78, 114.87, 112.80 (d, J = 7.6 Hz), 112.62, 112.03,102.02, 67.90, 67.25, 55.90, 39.85. ESI-MS m / z: 429.17 [M+H] + .

[0065] Example 14 Synthesis of Dg-3e, a substituted coumarin-eugenol derivative

[0066] The difference from Example 6 is that the coumarin intermediate in this example is 7-(3-bromopropoxy)-4-phenyl-2- H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-3e, yield: 81%.

[0067] 1 H NMR (600 MHz, CDCl3) δ 7.53 - 7.49 (m, 3H), 7.43 (dd, J = 6.1, 3.1Hz, 2H), 7.38 - 7.35 (m, 1H), 6.92 (d, J = 2.6 Hz, 1H), 6.86 - 6.83 (m, 1H), 6.81 - 6.78 (m, 1H), 6.73 - 6.69 (m, 2H), 6.21 (d, J = 1.9 Hz, 1H), 5.94(dddd, J = 16.7, 12.5, 7.5, 4.5 Hz, 1H), 5.06 (t, J = 12.9 Hz, 2H), 4.26 (t, J = 6.4 Hz, 2H), 4.20 (t, J= 6.3 Hz, 2H), 3.84 (d, J = 1.9 Hz, 3H), 3.33 (d, J = 6.7 Hz, 2H), 2.32 (p, J = 6.1 Hz, 2H). 13 C NMR (151 MHz, CDCl3) δ 162.18,161.30, 156.00, 155.83, 149.55, 146.52, 137.62, 135.63, 133.39, 129.58,128.83(2C), 128.40(2C), 127.95, 120.48, 115.68, 113.75, 112.66, 112.52,112.42, 111.86, 101.78, 65.55, 65.24, 55.90, 39.83, 29.11. ESI-MS m / z: 443.19 [M+H] + .

[0068] Example 15 Synthesis of Dg-4e, a substituted coumarin-eugenol derivative

[0069] The difference from Example 6 is that the coumarin intermediate in this example is 7-(4-bromobutoxy)-4-phenyl-2- H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crone-2-one compound. After the reaction was complete, water was added to dissolve cesium carbonate, and the pH was adjusted to approximately 7-8 with dilute hydrochloric acid. The aqueous layer was extracted three times with ethyl acetate, and the organic layers were combined. The organic layers were washed three times each with saturated NaCl and water (30 mL × 3), respectively. The organic layers were dried over anhydrous sodium sulfate, and the solvent was evaporated. Other parameters and procedures were as described in Example 6, yielding the substituted coumarin-eugenol derivative Dg-4e in 51% yield.

[0070] 1 H NMR (600 MHz, CDCl3) δ 7.53 - 7.50 (m, 3H), 7.45 - 7.42 (m, 2H),7.36 (d, J = 8.9 Hz, 1H), 6.89 (d, J = 2.5 Hz, 1H), 6.82 (d, J = 7.9 Hz, 1H), 6.78 (dt, J= 9.0, 1.5 Hz, 1H), 6.71 (d, J = 8.3 Hz, 2H), 6.21 (d, J = 0.9Hz, 1H), 5.99 - 5.92 (m, 1H), 5.11 - 5.02 (m, 2H), 4.13 (d, J = 5.9 Hz, 2H), 4.08 (q, J = 4.2 Hz, 2H), 3.85 (d, J = 0.9 Hz, 3H), 3.33 (d, J = 6.7 Hz, 2H), 2.03 (p, J = 3.2 Hz, 4H). 13 C NMR (151 MHz, CDCl3) δ 162.30, 161.34, 156.03,155.88, 149.43, 146.65, 137.67, 135.65, 133.07, 129.57, 128.83(2C), 128.40(2C), 127.93, 120.45, 115.65, 113.30, 112.75, 112.42, 112.35, 111.78, 101.60,68.68, 68.31, 55.92, 39.83, 25.98, 25.86. ESI-MS m / z: 457.20 [M+H] + .

[0071] Example 16 Synthesis of Dg-5e, a substituted coumarin-eugenol derivative

[0072] The difference from Example 6 is that the coumarin intermediate in this example is 7-(5-bromopentoxy)-4-phenyl-2- H -Creno-2-one replaced by 7-(4-bromobutoxy)-4-phenyl-2 H For other parameters and procedures of -chromene-2-one, refer to Example 6 to obtain the substituted coumarin-eugenol derivative Dg-5e, with a yield of 45%.

[0073] 1 H NMR (600 MHz, CDCl3) δ 7.52 - 7.50 (m, 3H), 7.45 - 7.42 (m, 2H), 7.38 - 7.35 (m, 1H), 6.87 (d, J= 2.6 Hz, 1H), 6.81 (dt, J = 8.2, 1.5 Hz, 1H), 6.77 (d, J = 8.7 Hz, 1H), 6.72 - 6.69 (m, 2H), 6.22 - 6.20 (m, 1H), 6.00- 5.91 (m, 1H), 5.09 - 5.04 (m, 2H), 4.06 - 4.01 (m, 4H), 3.85 - 3.84 (m,3H), 3.33 (d, J = 6.6 Hz, 2H), 1.91 (q, J = 7.1 Hz, 4H), 1.70 - 1.66 (m, 2H). 13C NMR (151 MHz, CDCl3) δ 162.32, 161.34, 156.03, 155.88, 149.42, 146.78,137.69, 135.65, 132.94, 129.58, 128.83(2C), 128.40(2C), 127.95, 120.47,115.63, 113.33, 112.69, 112.41, 112.39, 111.77, 101.62, 68.92, 68.44, 55.95,39.82, 28.98, 28.78, 22.65. ESI-MS m / z: 471.22 [M+H] + .

[0074] Example 17 Synthesis of Dg-2f, a substituted coumarin-eugenol derivative

[0075] The difference from Example 6 is that the coumarin intermediate in this example is 7-(2-bromoethoxy)-3-chloro-4-methyl-2- H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-2f, yield: 44%.

[0076] 1 H NMR (600 MHz, CDCl3) δ 7.52 (dd, J = 9.0, 2.0 Hz, 1H), 6.95 (dt, J= 9.0, 2.5 Hz, 1H), 6.91 - 6.87 (m, 2H), 6.74 - 6.71 (m, 2H), 5.95 (dddd, J =16.6, 7.8, 6.3, 2.2 Hz, 1H), 5.11 - 5.04 (m, 2H), 4.39 (d, J = 2.2 Hz, 4H), 3.84 (d, J = 2.1 Hz, 3H), 3.34 (d, J = 6.7 Hz, 2H), 2.55 - 2.53 (m, 3H). 13 CNMR (151 MHz, CDCl3) δ 161.69, 157.43, 153.04, 149.83, 147.95, 146.16,137.51, 134.25, 125.87, 120.54, 117.94, 115.79, 114.83, 113.52, 113.46,112.59, 101.71, 67.90, 67.33, 55.89, 39.84, 16.19. ESI-MS m / z: 401.11 [M+H] + .

[0077] Example 18 Synthesis of Dg-3f, a substituted coumarin-eugenol derivative

[0078] The difference from Example 6 is that the coumarin intermediate in this example is 7-(3-bromopropoxy)-3-chloro-4-methyl-2- H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H For other parameters and procedures of -chromene-2-one, refer to Example 6 to obtain the substituted coumarin-eugenol derivative Dg-3f, with a yield of 33%.

[0079] 1 H NMR (600 MHz, CDCl3) δ 7.50 (dt, J = 9.0, 1.5 Hz, 1H), 6.90 (dd, J = 8.9, 2.5 Hz, 1H), 6.86 - 6.82 (m, 2H), 6.73 - 6.68 (m, 2H), 5.94 (ddddd, J= 16.9, 8.2, 6.8, 4.2, 1.6 Hz, 1H), 5.11 - 5.02 (m, 2H), 4.25 (td, J = 6.2, 2.0 Hz, 2H), 4.19 (td, J = 6.1, 1.9 Hz, 2H), 3.85 - 3.82 (m, 3H), 3.32 (d, J = 6.7 Hz, 2H), 2.56 - 2.50 (m, 3H), 2.35 - 2.29 (m, 2H). 13 C NMR (151 MHz, CDCl3) δ 161.94, 157.48, 153.12, 149.53, 147.98, 146.48, 137.60, 133.42,125.83, 120.47, 117.76, 115.69, 113.72, 113.30, 113.27, 112.40, 101.43,65.49, 65.28, 55.89, 39.82, 29.09, 16.17. ESI-MS m / z: 415.13 [M+H] + .

[0080] Example 19 Synthesis of Dg-4f, a substituted coumarin-eugenol derivative

[0081] The difference from Example 6 is that the coumarin intermediate in this example is 7-(4-bromobutoxy)-3-chloro-4-methyl-2- H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-4f, yield: 59%.

[0082] 1 H NMR (600 MHz, CDCl3) δ 7.50 (dd, J = 8.8, 1.3 Hz, 1H), 6.89 (ddd, J = 8.9, 2.6, 1.3 Hz, 1H), 6.84 - 6.80 (m, 2H), 6.71 (d, J = 8.8 Hz, 2H), 5.96(ddtd, J= 16.8, 10.1, 6.7, 1.3 Hz, 1H), 5.12 - 5.04 (m, 2H), 4.15 - 4.05 (m,4H), 3.85 (d, J = 1.3 Hz, 3H), 3.33 (dd, J = 6.6, 1.9 Hz, 2H), 2.54 (d, J =1.3 Hz, 3H), 2.06 - 2.00 (m, 4H). 13 C NMR (151 MHz, CDCl3) δ 162.06, 157.49,153.15, 149.42, 148.01, 146.63, 137.66, 133.08, 125.82, 120.45, 117.68,115.66, 113.36, 113.29, 113.17, 112.35, 101.27, 68.67, 68.37, 55.91, 39.83,25.99, 25.84, 16.18. ESI-MS m / z: 429.15 [M+H] + .

[0083] Example 20 Synthesis of Dg-5f, a substituted coumarin-eugenol derivative

[0084] The difference from Example 6 is that the coumarin intermediate in this example is 7-(5-bromopentoxy)-3-chloro-4-methyl-2- H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-5f, yield: 59%.

[0085] 1 H NMR (600 MHz, CDCl3) δ 7.50 (d, J = 8.9 Hz, 1H), 6.88 (dd, J = 8.9,2.4 Hz, 1H), 6.82 - 6.78 (m, 2H), 6.72 - 6.68 (m, 2H), 5.95 (ddt, J = 16.8,9.9, 6.7 Hz, 1H), 5.10 - 5.03 (m, 2H), 4.03 (dt, J= 8.8, 6.5 Hz, 4H), 3.84(s, 3H), 3.32 (d, J = 6.7 Hz, 2H), 2.53 (s, 3H), 1.93 - 1.87 (m, 4H), 1.70 -1.66 (m, 2H). 13 C NMR (151 MHz, CDCl3) δ 162.08, 157.49, 153.13, 149.41,148.02, 146.76, 137.68, 132.94, 125.84, 120.46, 117.65, 115.63, 113.30 (d, J = 4.4 Hz), 113.15, 112.38, 101.28, 68.90, 68.50, 55.94, 39.82, 28.97, 28.76,22.64, 16.17. ESI-MS m / z: 443.16 [M+H] + .

[0086] Example 21 Synthesis of Dg-2g, a substituted coumarin-eugenol derivative

[0087] The difference from Example 6 is that the coumarin intermediate in this example is 7-(2-bromoethoxy)-2-oxo-2- H -Ethyl chromene-3-carboxylate instead of 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the coumarin-eugenol derivative Dg-2g, yield: 34%.

[0088] 1 H NMR (600 MHz, CDCl3) δ 8.49 (s, 1H), 7.49 (dd, J = 8.7, 1.5 Hz, 1H), 6.94 (dt, J = 8.8, 2.0 Hz, 1H), 6.90 - 6.87 (m, 2H), 6.74 - 6.70 (m,2H), 5.99 - 5.91 (m, 1H), 5.10 - 5.05 (m, 2H), 4.42 - 4.37 (m, 6H), 3.84 (d, J = 1.7 Hz, 3H), 3.33 (d, J = 6.7 Hz, 2H), 1.39 (td,J = 7.2, 1.6 Hz, 3H). 13 CNMR (151 MHz, CDCl3) δ 164.27, 163.45, 157.45, 157.15, 149.84, 148.94,146.09, 137.49, 134.33, 130.69, 120.54, 115.80, 114.86, 114.28, 114.11,112.60, 111.85, 101.28, 67.83, 67.54, 61.73, 55.87, 39.84, 14.29. ESI-MS m / z:425.16 [M+H] + .

[0089] Example 22 Synthesis of Dg-3g, a substituted coumarin-eugenol derivative

[0090] The difference from Example 6 is that the coumarin intermediate in this example is 7-(3-bromopropoxy)-2-oxo-2- H -Ethyl chromene-3-carboxylate instead of 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-3g, yield: 31%.

[0091] 1 H NMR (600 MHz, CDCl3) δ 8.49 (s, 1H), 7.48 (dd, J = 8.7, 1.4 Hz, 1H), 6.89 (dt, J = 8.7, 1.9 Hz, 1H), 6.86 – 6.81 (m, 2H), 6.74 – 6.68 (m,2H), 5.99 – 5.90 (m, 1H), 5.10 – 5.03 (m, 2H), 4.40 (qd, J = 7.1, 1.5 Hz, 2H), 4.29 (td, J = 6.1, 1.5 Hz, 2H), 4.19 (td, J = 6.0, 1.4 Hz, 2H), 3.84 (d, J = 1.5 Hz, 3H), 3.35 – 3.30 (m, 2H), 2.32 (q, J= 6.0 Hz, 2H), 1.40 (td, J =7.1, 1.4 Hz, 3H). 13 C NMR (151 MHz, CDCl3) δ 164.53, 163.49, 157.55, 157.20,149.53, 148.99, 146.42, 137.59, 133.48, 130.69, 120.46, 115.70, 114.05,113.96, 113.74, 112.39, 111.62, 101.00, 65.50, 65.37, 61.70, 55.87, 39.82,29.01, 14.30. ESI-MS m / z: 439.18 [M+H] + .

[0092] Example 23 Synthesis of the substituted coumarin-eugenol derivative Dg-4g

[0093] The difference from Example 6 is that the coumarin intermediate in this example is 7-(4-bromobutoxy)-2-oxo-2- H -Ethyl chromene-3-carboxylate instead of 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-4g, yield: 35%.

[0094] 1 H NMR (600 MHz, CDCl3) δ 8.49 (s, 1H), 7.47 (d, J = 8.7 Hz, 1H), 6.88– 6.79 (m, 3H), 6.70 (d, J = 8.4 Hz, 2H), 5.95 (dq, J = 16.6, 7.4 Hz, 1H), 5.06 (t, J = 13.1 Hz, 2H), 4.39 (q, J = 7.2 Hz, 2H), 4.14 (d, J = 6.3 Hz, 2H), 4.09 – 4.05 (m, 2H), 3.84 (s, 3H), 3.32 (d, J = 6.7 Hz, 2H), 2.03 (q, J= 6.5 Hz, 4H), 1.39 (t, J = 7.2 Hz, 3H). 13 C NMR (151 MHz, CDCl3) δ 164.67,163.52, 157.59, 157.22, 149.41, 149.02, 146.59, 137.64, 133.11, 130.67,120.44, 115.66, 114.01, 113.95, 113.28, 112.33, 111.52, 100.84, 68.64(2C),61.69, 55.89, 39.82, 25.96, 25.79, 14.30. ESI-MS m / z: 453.19 [M+H] + .

[0095] Example 24 Synthesis of Dg-5g, a substituted coumarin-eugenol derivative

[0096] The difference from Example 6 is that the coumarin intermediate in this example is 7-(5-bromopentoxy)-2-oxo-2- H -Creptene-3-carboxylate instead of 7-(2-bromoethoxy)-2-oxo-2 H -chromene-3-carboxylate, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-5g, yield: 31%.

[0097] 1 H NMR (600 MHz, DMSO- d 6 )) δ 8.72 (s, 1H), 7.83 (d, J = 8.7 Hz, 1H),7.03 – 6.99 (m, 2H), 6.86 (d, J = 8.1 Hz, 1H), 6.77 (d, J = 2.0 Hz, 1H), 6.66(dd, J = 8.2, 2.0 Hz, 1H), 5.93 (ddt, J = 16.8, 10.0, 6.7 Hz, 1H), 5.09 –5.01 (m, 2H), 4.28 (q, J = 7.1 Hz, 2H), 4.15 (t, J= 6.4 Hz, 2H), 3.93 (t, J = 6.4 Hz, 2H), 3.73 (s, 3H), 3.28 (d, J = 6.8 Hz, 2H), 1.82 (p, J = 6.8 Hz, 2H), 1.76 (q, J = 7.0 Hz, 2H), 1.57 (qd, J = 8.8, 6.2 Hz, 2H), 1.31 (t, J =7.1 Hz, 3H). 13 C NMR (151 MHz, DMSO- d 6 )) δ 164.70, 163.32, 157.48, 156.77,149.68, 149.46, 146.92, 138.45, 132.76, 132.12, 120.71, 115.94, 114.10,113.91, 113.64, 113.04, 111.80, 101.15, 69.08, 68.71, 61.39, 55.93, 39.55,28.95, 28.56, 22.63, 14.59. ESI-MS m / z: 467.20 [M+H] + .

[0098] Example 25 Synthesis of Dg-2i, a substituted coumarin-eugenol derivative

[0099] The difference from Example 6 is that the coumarin intermediate in this example is 7-(2-bromoethoxy)-4-(trifluoromethyl)-2 H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-2i, yield: 29%.

[0100] 1 H NMR (600 MHz, CDCl3) δ 7.64 - 7.61 (m, 1H), 6.99 - 6.96 (m, 2H), 6.88 (d, J = 8.0 Hz, 1H), 6.74 - 6.71 (m, 2H), 6.62 (s, 1H), 5.95 (ddt,J =16.9, 10.1, 6.7 Hz, 1H), 5.11 - 5.05 (m, 2H), 4.43 - 4.38 (m, 4H), 3.85 (s,3H), 3.34 (d, J = 6.7 Hz, 2H). 13 C NMR (151 MHz, CDCl3) δ 162.70, 159.41,156.26, 149.83, 146.10, 141.58 (d, J = 32.9 Hz), 137.49, 134.32, 126.31 (d, J = 2.5 Hz), 120.54, 115.80, 114.83, 113.88, 112.60, 112.41 (d, J = 5.8 Hz),107.27, 102.35, 67.84, 67.47, 55.86, 39.84. ESI-MS m / z: 421.13[M+H] + .

[0101] Example 26 Synthesis of Dg-3i, a substituted coumarin-eugenol derivative

[0102] The difference from Example 6 is that the coumarin intermediate in this example is 7-(3-bromopropoxy)-4-(trifluoromethyl)-2 H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-3i, yield: 29%.

[0103] 1 H NMR (600 MHz, DMSO- d 6 ) δ 7.61 (dd, J = 9.0, 2.1 Hz, 1H), 7.16 (d, J = 2.5 Hz, 1H), 7.07 (dd, J = 9.0, 2.5 Hz, 1H), 6.90 (d, J = 8.1 Hz, 1H), 6.84(s, 1H), 6.77 (d, J = 2.0 Hz, 1H), 6.66 (dd,J = 8.2, 2.0 Hz, 1H), 5.92 (ddt, J = 16.7, 9.7, 6.8 Hz, 1H), 5.09 - 4.99 (m, 2H), 4.28 (t, J = 6.3 Hz, 2H), 4.08 (t, J = 6.2 Hz, 2H), 3.73 (s, 3H), 3.28 (d, J = 6.8 Hz, 2H), 2.18 (p, J = 6.2 Hz, 2H). 13 C NMR (151 MHz, DMSO- d 6 ) δ 162.83, 159.21, 156.35, 149.55,146.66, 140.02, 138.41, 133.16, 126.32, 120.72, 115.97, 114.25, 114.14,113.74, 113.05, 106.88, 102.65, 65.90, 65.45, 55.93, 40.52, 28.95. ESI-MS m / z: 435.14 [M+H] + .

[0104] Example 27 Synthesis of Dg-4i, a substituted coumarin-eugenol derivative

[0105] The difference from Example 6 is that the coumarin intermediate in this example is 7-(4-bromobutoxy)-4-(trifluoromethyl)-2 H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-4i, yield: 29%.

[0106] 1 H NMR (600 MHz, CDCl3) δ 7.60 (d, J = 8.9 Hz, 1H), 6.93 - 6.86 (m,2H), 6.81 (d, J = 7.9 Hz, 1H), 6.71 (d, J = 8.6 Hz, 2H), 6.61 (s, 1H), 5.96(td,J = 16.7, 6.8 Hz, 1H), 5.12 - 5.02 (m, 2H), 4.15 (t, J = 5.8 Hz, 2H), 4.08 (t, J = 5.6 Hz, 2H), 3.85 (s, 3H), 3.33 (d, J = 6.7 Hz, 2H), 2.06 - 2.00(m, 4H). 13 C NMR (151 MHz, CDCl3) δ 163.05, 159.50, 156.38, 149.41, 146.60,141.63 (q, J = 32.7 Hz), 137.64, 133.12, 126.27 (d, J = 2.7 Hz), 120.45,115.66, 113.77, 113.28, 112.34, 112.09 (q, J = 5.9 Hz), 106.91, 101.87,68.64, 68.54, 55.89, 39.82, 25.98, 25.79. ESI-MS m / z: 449.16 [M+H] + .

[0107] Example 28 Synthesis of Dg-5i, a substituted coumarin-eugenol derivative

[0108] The difference from Example 6 is that the coumarin intermediate in this example is 7-(5-bromopentoxy)-4-(trifluoromethyl)-2 H -Crone-2-one replaced by 4-(3-bromopropoxy)-2 H -Crotene-2-one, other parameters and operations are as described in Example 6, to obtain the substituted coumarin-eugenol derivative Dg-5i, yield: 32%.

[0109] 1 H NMR (600 MHz, DMSO- d 6 ) δ 7.60 (dd, J = 9.0, 2.3 Hz, 1H), 7.13 (d, J = 2.5 Hz, 1H), 7.05 (dd, J= 9.1, 2.5 Hz, 1H), 6.87 - 6.82 (m, 2H), 6.77 (d, J = 2.0 Hz, 1H), 6.66 (dd, J = 8.2, 2.0 Hz, 1H), 5.93 (ddt, J = 16.8, 9.9,6.7 Hz, 1H), 5.09 - 5.00 (m, 2H), 4.14 (t, J = 6.4 Hz, 2H), 3.93 (t, J = 6.4Hz, 2H), 3.73 (s, 3H), 3.28 (d, J = 6.8 Hz, 2H), 1.79 (dp, J = 31.0, 6.8 Hz, 4H), 1.60 - 1.54 (m, 2H). 13 C NMR (151 MHz, DMSO- d 6 ) δ 163.01, 159.21, 156.37,149.47, 146.93, 139.84 (t, J = 32.1 Hz), 138.44, 132.75, 126.25 (d, J = 2.4Hz), 120.69, 115.92, 114.15, 113.90, 113.51 (q, J = 5.7 Hz), 113.03, 106.72,102.56, 69.00, 68.70, 55.92, 39.55, 28.95, 28.54, 22.61. ESI-MS m / z: 463.17[M+H] + .

[0110] Experimental Example 1: Inhibitory effect of substituted coumarin-eugenol derivatives on monoamine oxidase-B The inhibitory activity of the substituted coumarin-eugenol derivatives obtained in Examples 1-28 against monoamine oxidase-B (MAO-B, Sigma) was determined by fluorescence spectrophotometry. Results are expressed as inhibition rates, with ladostigil as a positive control. All tests were performed on a PowerWave XS2 full-wavelength microplate reader, with absorbance measured at 490 nm. The test concentration of the compound was 40 μM, and the inhibition rate was calculated using the following formula: Inhibition rate (%) = [1 - (sample - sample background) / (blank group - blank background)] × 100%. In the blank group, 10 µL of PBS (pH=7.6) replaced 10 µL of sample solution; in the blank background group, 30 µL of PBS (pH=7.6) replaced 30 µL of substrate; in the sample background group, 10 µL of PBS (pH=7.6) replaced 10 µL of sample solution; and in the sample background group, 30 µL of PBS (pH=7.6) replaced 30 µL of substrate.

[0111] The experimental results are shown in Table 1.

[0112] Table 1. Inhibitory activity of substituted coumarin-eugenol derivatives against MAO-B ; As shown in Table 1, all compounds obtained in this invention exhibit varying degrees of inhibitory activity against MAO-B. Among them, compound Dg-4d showed the strongest inhibitory activity, reaching 79.8%, which is more potent than the positive control ladostigil. This demonstrates that the substituted coumarin-eugenol derivatives obtained in this invention can be used to prepare anti-Alzheimer's drugs based on MAO-B inhibition.

[0113] Additionally, based on the compound structure and the overall structure-activity relationship in Table 1, the following may be observed: two methyl substitutions on the aromatic ring of the coumarin moiety result in better activity; eugenol linkers attached to the 7th position of the coumarin core show better activity than those attached to the 4th position; introducing larger groups at the 3rd and 4th positions of the coumarin core significantly reduces activity; the chain length between the coumarin and eugenol moieties has a significant impact on activity, with a carbon chain length of 4 being the most suitable.

[0114] Experimental Example 2: Inhibition of Aβ by substituted coumarin-eugenol derivatives 1-42 Self-aggregation The inhibition of Aβ by the substituted coumarin-eugenol derivatives obtained in Examples 1-28 was determined using the thiosulfate T (ThT) fluorescence method. 1-42 Self-aggregation activity, compounds and Aβ 1-42 The final concentration of all samples was 20 μM. Curcumin, resveratrol, and 7-hydroxycoumarin were used as positive controls.

[0115] Take 10 μL of 40 μM Aβ respectively 1-42The protein was mixed with 10 μL of a 40 μM compound and incubated at 37°C for 48 h. The blank control was 10 μL of 40 μM Aβ. 1-42 The protein was mixed with 10 μL of pH 7.4 phosphate buffer and incubated together; the positive control was Aβ. 1-42 The protein was co-incubated with resveratrol. After 72 h, the incubation solution was transferred to black 96-well plates, and 180 μL of 5 μM thiamine T solution was added. The plates were then incubated in the dark at room temperature for 5 min. Finally, the fluorescence absorbance was measured using a multi-mode microplate reader, with an excitation wavelength of 450 nm and an absorption wavelength of 485 nm. The Aβ value in the negative control experiment was used as a reference. 1-42 Using the fluorescence intensity of the compound bound to thiosulfate T as a control, the effect of the compound on Aβ was determined. 1-42 Inhibition rate of protein aggregation. The results are shown in Table 2.

[0116] Table 2. Effects of substituted coumarin-eugenol derivatives on Aβ 1-42 Inhibitory activity of self-aggregation ; The results show that the compound described in this invention has an effect on Aβ. 1-42 These compounds exhibit strong inhibitory activity, with most showing better activity than 7-hydroxycoumarin and curcumin. Compounds Dg-4d and Dg-5d demonstrated the strongest inhibitory activity, reaching 92.6% and 90.4%, respectively. This demonstrates that the substituted coumarin-eugenol derivatives described in this invention have significant development potential and can be used to inhibit Aβ. 1-42 Self-aggregation, used to prepare drugs for treating Alzheimer's disease.

[0117] Furthermore, based on the compound structure and the overall structure-activity relationship in Table 2, the following may be observed: electron-donating substitutions on the aromatic ring of the coumarin moiety have higher activity than electron-withdrawing substitutions; simultaneously, phenyl substitutions on the coumarin core have better activity than other electron-withdrawing substitutions; eugenol linkers attached to the 7th position of the coumarin core have better activity than those attached to the 4th position; additionally, the chain length between the coumarin moiety and eugenol affects the activity, with a carbon chain length of 4 being more desirable.

[0118] Experimental Example 3: In vitro antioxidant activity experiment of substituted coumarin-eugenol derivatives The in vitro antioxidant activity of the substituted coumarin-eugenol derivatives obtained in Examples 1-28 was determined by oxygen radical uptake capacity (ORAC) method. AAPH was used as the source of peroxy radicals, and sodium fluorescein (FL) was used as the fluorescent indicator to evaluate the antioxidant capacity of some compounds. The experimental results were expressed as Trolox equivalents.

[0119] Different concentrations of the compound or 20 μL of Trolox and 120 μL of FL dilution were respectively pipetted into black 96-well culture plates, mixed with a pipette, and incubated at 37 ℃ for 15 min. Then, 60 μL of AAPH was quickly added. Fluorescence values ​​were measured and recorded every 1 min using a multi-mode microplate reader. The excitation wavelength was 485 nm and the emission wavelength was 535 nm, for a total of 240 min. The blank control was tested with 20 μL of PBS instead of the compound. The area between the curve and the coordinate axis (AUC) was calculated by integration using ORIGIN software. The formula for calculating the protective area of ​​the sample is: Net AUC = AUC antioxidant – AUC blank. The ORAC-FL value was calculated as: [(AUC Sample - AUC blank) / (AUC Trolox - AUC blank)] / [Trolox concentration / sample concentration). The sample ORAC value was expressed as a Trolox equivalent.

[0120] Table 3. In vitro antioxidant activity of substituted coumarin-eugenol derivatives ; As shown in Table 3, some of the compounds obtained in this invention exhibit good antioxidant activity in vitro. Among them, compounds Dg-4d and Dg-3d have ORAC values ​​of 5.8 and 5.2 at a concentration of 5 μM, respectively, indicating good antioxidant properties. This demonstrates that the substituted coumarin-eugenol derivatives obtained in this invention can be used to prepare drugs for treating Alzheimer's disease based on their antioxidant properties.

[0121] Experimental Example 4: Inhibitory effect of substituted coumarin-eugenol derivatives on acetylcholinesterase The inhibitory effect of the substituted coumarin-eugenol derivatives obtained in Examples 1-28 on acetylcholinesterase was determined using the Ellman method. Results were expressed as inhibition rates, with tacrine used as a positive control. All tests were performed on a PowerWave XS2 full-wavelength microplate reader at 37 °C. Data analysis was performed using Origin software.

[0122] In a 96-well plate, six wells were selected, and 10 μL of enzyme solution and 0, 5, 10, 20, 35, and 50 μL of the test compound solution were added to each well. 0.1 mol / L pH 8.0 phosphate buffer was added to bring the total volume to 100 μL. The plate was incubated at 37 ℃ for 15 min using a full-wavelength microplate reader. Immediately afterwards, a mixture of 10 μL ATC solution, 10 μL DTNB solution, and 80 μL phosphate buffer was added to a total volume of 100 μL. The absorbance was measured at λ=412 nm for 2 min. The experimental results are shown in Table 4. The results indicate that some of the compounds described in this invention have good inhibitory activity against acetylcholinesterase, which is better than that of the parent nucleus 7-hydroxycoumarin. Among them, compounds Dg-4d, Dg-3i, Dg-4i, and Dg-5i showed the best inhibition of acetylcholinesterase activity, with inhibition rates of 75.5%, 73.2%, 74.3%, 83.6%, and 84.3%, respectively. They exhibited good inhibitory activity and could be used to prepare drugs for treating Alzheimer's disease.

[0123] Table 4. Inhibitory activity of substituted coumarin-eugenol derivatives against acetylcholinesterase ; Experimental Example 5: Metal Complexation Experiment of Substituted Coumarin-Eugenol Derivatives The metal complexing ability of the substituted coumarin-eugenol derivative Dg-4d obtained in Example 11 was determined by UV-vis method, and the specific steps are as follows: 1. Solution preparation: (1) Compound Dg-4d solution: Weigh a certain amount of the compound and prepare it to 1 mM with anhydrous ethanol.

[0124] (2) Metal ion solution: Weigh a certain amount of NaCl, KCl, CaCl2, ZnSO4, CuSO4 and FeSO4 and prepare 10 mM solution with ultrapure water, then dilute to 1 mM with anhydrous ethanol.

[0125] 2. The interaction of compound Dg-4d with NaCl, KCl, CaCl2, ZnSO4, CuSO4, and FeSO4. Take six 5 mL centrifuge tubes and add 40 μL of 1 mM Dg-4d compound solution to each. Then add 40 μL of 1 mM NaCl, KCl, CaCl2, ZnSO4, CuSO4, and FeSO4 solutions respectively. Finally, add anhydrous ethanol to bring the total volume to 4000 μL, ensuring that the final concentration of both compound Dg-4g and the metal ions is 10 μM. For the blank control, add anhydrous ethanol to 40 μL of 1 mM compound solution until the concentration is the same as in the sample. After mixing, let stand at room temperature for 30 min, then pour into a quartz cuvette and scan the absorption curve using a UV-Vis spectrometer. The test temperature is room temperature, the test range is 200–700 nm, the wavelength interval is 1 nm, the scan rate is 200 nm / min, and each sample is tested three times, and the average value is taken.

[0126] See results Figure 1 ,Depend on Figure 1 It is evident that the compound Dg-4d of this invention possesses strong metal complexing ability and exhibits strong affinity for Cu. 2+ It exhibits good selectivity; other substituted coumarin-eugenols with similar structures also have similar effects, proving that the substituted coumarin-eugenol derivatives obtained in this invention can be used to prepare drugs for treating Alzheimer's disease based on metal complexation.

[0127] Experimental Example 6: Toxicity Study of Substituted Coumarin-Eugenol Derivatives on Nerve Cells The toxicity of the substituted coumarin-eugenol derivatives obtained in Examples 1-28 to nerve cells (SH-SY5Y) was determined by the MTT assay.

[0128] Cell viability (%) in each sample = (OD sample - OD blank) / (OD control - OD blank) × 100%; Cell inhibition rate (%) in each sample = 100% - Cell viability (%) in each sample. Plot the inhibition rate against concentration; the concentration with an inhibition rate of 50% is the IC50 of the compound. 50 Values. See Table 5 for the results.

[0129] Table 5. Toxicity of substituted coumarin-eugenol derivatives on nerve cells (SH-SY5Y) ; As shown in Table 5, the IC50 values ​​of SH-SY5Y cells containing compounds Dg-4d and Dg-5d, which exhibit the strongest inhibitory activity against MAO-B and Aβ self-aggregation, are... 50 The values ​​were 121.4 μM and 120.1 μM, respectively, which are greater than 100 μM, indicating low neurotoxicity and good safety.

[0130] Example 7: Study on the neuroprotective effects of substituted coumarin-eugenol derivatives Dg-4d and Dg-5d on H2O2-induced SH-SY5Y cell damage. The neuroprotective effects of the substituted coumarin-eugenol derivatives Dg-4d and Dg-5d obtained in Examples 11 and 12 on H2O2-induced SH-SY5Y cell damage were determined using the MTT assay. The specific steps are as follows: SH-SY5Y cells were passaged in 96-well plates at a cell density of 1 × 10⁻⁶ cells per well. 4 After removing the culture medium, different concentrations of compounds Dg-4d and Dg-5d (1, 5, and 10 μM) were added and incubated for 3 h, with 7-hydroxycoumarin as a positive control. Then, 10 μL of H2O2 (200 μM) was added and incubation continued for 12 h. Cell viability was determined using the MTT assay as described in Example 6 above, and the results are shown below. Figure 2 .

[0131] Depend on Figure 2 It is evident that the substituted coumarin-eugenol derivatives Dg-4d and Dg-5d obtained in this invention have good neuroprotective effects against H2O2-induced SH-SY5Y cell damage, and exhibit better protective ability than 7-hydroxycoumarin; among them, the cell survival rate of SH-SY5Y cells increased to 89.3% after treatment with 10 μM compound Dg-4d.

[0132] Experimental Example 8: The effect of substituted coumarin-eugenol derivative Dg-4d on Aβ 1-42 Study on the effect of induced memory impairment in mice The passive avoidance experiment was used to evaluate the effect of the substituted coumarin-eugenol derivative Dg-4d obtained in Example 11 on Aβ. 1-42 The study investigated the effect of induced memory impairment in mice. The specific steps are as follows: (1) Establishment of AD mouse model Sixty male SD mice (3 months old) were used and placed in an environment with a temperature of 22-25 °C, relative humidity of 50-70%, and a light-dark cycle of 12 h. The 60 male SD mice were randomly divided into 6 groups: a) control group (injected with physiological saline and administered distilled water by gavage); b) AD model group (Aβ... 1-42 (c) High-dose group (Dg-4d-H 8.0 mg / kg); (d) Medium-dose group (Dg-4d-M 4.0 mg / kg); (e) Low-dose group (Dg-4d-L 2.0 mg / kg). Aβ 1-42Forty-eight hours after injection, each treatment group was administered distilled water once daily for 14 consecutive days. The control group and AD model group were administered distilled water by gavage using the same method. After 14 days of treatment, a passive avoidance test was conducted as follows.

[0133] (2) Passive avoidance experiment In vivo memory enhancement was assessed using a passive avoidance experiment in mice, comprising two separate tests (a training test and a test 24 hours later). The experimental setup consisted of two identical compartments (a light compartment and a dark compartment) separated by a guillotine door, illuminated by 250 lx LED lights. In the training test, mice were first placed in the light chamber and allowed free movement for 5 minutes to familiarize themselves with their environment. The door was then opened, and the mice quickly entered the dark compartment while an electrical stimulator was activated, delivering an electric shock (24V, 0.5mA) to the animal's paws. This training was repeated for 5 minutes. Mice that failed to enter the dark compartment within 180 seconds were eliminated and subjected to the test.

[0134] The training experiment was conducted 24 hours later. Mice were placed back into the light chamber and the door was opened. The total test time was set to 5 minutes. During the test time, latency and the number of errors were recorded. Latency (i.e., the time it takes for a mouse to enter the dark chamber) was defined as the time it takes for the mouse to enter the dark chamber, and the number of errors was the number of times the mouse entered the dark chamber within 5 minutes. Results are shown below. Figure 3 .

[0135] Depend on Figure 3 As shown, Aβ 1-42 The latency in the treatment group (model group, 98 sec) was significantly shorter than that in the control group (232 sec), and the average number of errors within 5 minutes was significantly increased (5.3 in the model group, 2.0 in the control group), indicating that Aβ injection... 1-42 It significantly induced memory impairment in mice. Treatment with donepezil and different concentrations of compound Dg-4d (2.0, 4.0, 8.0 mg / kg) significantly reversed latency and mean error count compared to the model group. Furthermore, compound Dg-4d prolonged latency and reduced error count in a dose-dependent manner. Specifically, the 8.0 mg / kg dose of compound Dg-4d showed better efficacy in increasing latency and reducing error count compared to the donepezil group.

[0136] As can be seen from the above, the substituted coumarin-eugenol derivatives obtained in this invention possess good MAO-B inhibitory activity, inhibition of Aβ self-aggregation, inhibition of acetylcholinesterase, antioxidant activity, and metal chelating effect, and exhibit low toxicity to nerve cells, demonstrating high safety. Among them, compound Dg-4d exhibits good Aβ... 1-42 Induced SH-SY5Y cell damage has a strong neuroprotective effect; animal experiments show that high-dose compound Dg-4d can effectively improve Aβ.1-42 The resulting memory impairment in mice demonstrates that the compounds provided by this invention are highly suitable for preparing drugs against Alzheimer's disease.

[0137] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A substituted coumarin-eugenol derivative, characterized in that, having a structure as shown in general formula (I) or general formula (II): wherein R1 is selected from hydrogen, methyl, chlorine or formyloxyethyl; R2 is selected from hydrogen, methyl, trifluoromethyl or phenyl; and n is an integer from 2 to 5.

2. The substituted coumarin-eugenol derivative according to claim 1, characterized in that, The derivative is any one of the following compounds: 。 3. The substituted coumarin-eugenol derivative of any one of claims 1 or 2 in a pharmaceutically acceptable salt.

4. A process for the preparation of a substituted coumarin-eugenol derivative as claimed in any one of claims 1 or 2, characterized in that, comprising the following steps: S1. subjecting the first reactant or the second reactant to a monosubstitution reaction with Br-(CH2) n - Br in a basic environment to obtain the first intermediate product or the second intermediate product; S2. performing a nucleophilic substitution reaction of the first intermediate product or the second intermediate product obtained in step S1 with eugenol under alkaline conditions to obtain the substituted coumarin-eugenol derivative of general formula (I) or general formula (II); The structure of the first reactant is The structure of the second reactant is wherein R1 is selected from hydrogen, methyl, chloro or formyloxyethyl; R2 is selected from hydrogen, methyl, trifluoromethyl or phenyl, and n is an integer from 2 to 5.

5. The method of claim 4, wherein, In step S1, the alkaline conditions are provided by cesium carbonate, potassium carbonate or sodium carbonate; and / or, in step S2, the alkaline conditions are provided by cesium carbonate or potassium carbonate; and / or, the solvent in step S2 is N,N-dimethylformamide or acetonitrile.

6. The method of claim 4, wherein, In step S1, the mono-substitution reaction is performed at 25-60 °C for 2-8 h; and / or, in step S2, the nucleophilic substitution reaction is performed at 25-100 °C for 2-8 h.

7. Use of the substituted coumarin-eugenol derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 or 2 in the manufacture of a medicament, characterized in that, The drug is used for inhibiting monoamine oxidase-B, inhibiting acetylcholinesterase, inhibiting Aβ protein aggregation, antioxidation and / or chelating metal ions.

8. Use of the substituted coumarin-eugenol derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 or 2 in the manufacture of a medicament, characterized in that, The drug is used for preventing and / or treating neurodegenerative diseases.

9. Use according to claim 7, characterized in that, The neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, cerebrovascular dementia, Huntington's disease, amyotrophic lateral sclerosis or myasthenia gravis.

10. A pharmaceutical preparation, characterized in that, A pharmaceutical composition comprising a therapeutically effective amount of the substituted coumarin-eugenol derivative of any one of claims 1 or 2, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients.

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