Preparation method and application of alkoxy-substituted chalcone derivative

Abstract

The invention discloses a preparation method and application of an alkoxy substituted chalcone derivative, and belongs to the field of medicines, the alkoxy substituted chalcone derivative refers to a compound (E)-3-(3, 4-dihydroxyphenyl)-1-(2, 5-dimethoxyphenyl) prop-2-ene-1-ketone, and the structural formula is shown in the specification. The preparation method comprises the following steps: adding 3, 5-dimethoxyacetophenone into a reaction container, adding absolute ethyl alcohol for dissolving, adding a 60% sodium hydroxide aqueous solution, stirring at room temperature, adding 3, 4-dihydroxybenzaldehyde, stirring at 70-90 DEG C for reaction, cooling to room temperature after the reaction is finished, acidifying with hydrochloric acid, extracting with ethyl acetate, drying with anhydrous sodium sulfate, and concentrating under reduced pressure to obtain the 3, 5-dimethoxyacetophenone. And purifying through silica gel column chromatography to obtain a product. The invention also provides application of the alkoxy-substituted chalcone derivative in preparation of drugs for inhibiting oxidative stress injury of nerve cells and application of the alkoxy-substituted chalcone derivative in preparation of drugs for treating neurodegenerative diseases. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of medicine, and specifically relates to a preparation method and application of an alkoxy substituted chalcone derivative. Background Art

[0002] Neurodegenerative diseases are a class of chronic diseases characterized by degeneration and death of neurons in the central nervous system, mainly including Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), etc. These diseases usually have the following common characteristics: complex pathogenesis, involving genetic factors, environmental factors, oxidative stress, neuroinflammation, etc.; slow progression, it may take years or even decades from the onset of early symptoms to severe functional impairment; difficult to treat, current treatment methods mainly focus on relieving symptoms, and there is no cure.

[0003] Chalcone is a class of compounds with a wide range of biological activities. Its structure contains conjugated double bonds and carbonyl groups, which can interact with a variety of biological targets. In recent years, chalcone derivatives have shown great potential in the treatment and diagnosis of neurodegenerative diseases. The main research directions include:

[0004] (1) Inhibition of β-amyloid protein (Aβ) aggregation: Aβ aggregation is one of the key pathological features of Alzheimer's disease (AD). Studies have shown that chalcone derivatives can inhibit Aβ aggregation by binding to it, thereby reducing neurotoxicity. For example, some chalcone derivatives are designed to have groups that specifically bind to Aβ and can effectively inhibit the formation of Aβ fibers (Chen Yiping, Wang Ling, Xu Jun, Huang Zhishu, Gu Lianquan. Molecular dynamics mechanism of chalcone derivatives inhibiting beta-Amyloid aggregation [C] / / National Medicinal Chemistry Academic Conference and the Fourth Sino-British Medicinal Chemistry Academic Conference, 2013.).

[0005] (2) Antioxidant and anti-inflammatory effects: Oxidative stress and neuroinflammation are important pathological mechanisms of neurodegenerative diseases. Chalcone derivatives have good antioxidant activity due to the conjugated double bonds and carbonyl groups in their structures, which can scavenge free radicals and reduce oxidative stress damage. In addition, some derivatives also show anti-inflammatory effects, which help to relieve neuroinflammation.

[0006] (3) Multi-target drug development: In view of the complexity of neurodegenerative diseases, multi-target drugs have become a research hotspot. Chalcone derivatives can act on multiple targets simultaneously, such as acetylcholinesterase (AChE), monoamine oxidase B (MAO-B), etc. For example, some chalcone derivatives are designed as dual inhibitors of AChE and MAO-B, which can both increase the level of neurotransmitters and reduce the production of neurotoxic substances (Chen Lin, Wang Keren, Sang Zhipei. Research progress of chalcone derivatives in the treatment and diagnosis of Alzheimer's disease [J]. Pharmaceutical Progress, 2019, 43(4): 293-299.).

[0007] (4) Development of diagnostic probes: Chalcone derivatives can also be used as diagnostic probes for early detection of neurodegenerative diseases. For example, some chalcone derivatives have been developed as fluorescent probes that can specifically bind to Aβ plaques and are used for early diagnosis of Alzheimer's disease (Chen Lin, Wang Keren, Sang Zhipei. Research progress of chalcone derivatives in the treatment and diagnosis of Alzheimer's disease [J]. Pharmaceutical Progress, 2019, 43(4): 293-299.).

[0008] At present, significant progress has been made in the research of chalcone derivatives in the treatment and diagnosis of neurodegenerative diseases. Through structural optimization and multi-target design, chalcone derivatives have shown good biological activity and clinical application potential. However, further clinical trials are still needed to verify their safety and efficacy in order to promote their widespread application in neurodegenerative diseases. Summary of the invention

[0009] The purpose of the present invention is to provide a preparation method and use of an alkoxy substituted chalcone derivative. The present invention uses an oxidative stress model of nerve cells to study the role of alkoxy substituted chalcone derivatives in neuroprotection, providing an important new idea and potential therapeutic strategy for the treatment of neurodegenerative diseases.

[0010] The alkoxy-substituted chalcone derivative of the present invention is specifically the compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one, English name (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one, CAS: 152809-88-6, chemical formula C 17 H 16 O 5 , molecular weight 300.31, structural formula:

[0011]

[0012] The preparation method of the compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one of the present invention comprises the following steps:

[0013] Add 3,5-dimethoxyacetophenone to a reaction container, add anhydrous ethanol to dissolve, add 60% sodium hydroxide aqueous solution, stir at room temperature, add 3,4-dihydroxybenzaldehyde, stir and react at 70°C-90°C, cool to room temperature after the reaction, acidify with hydrochloric acid, extract with ethyl acetate, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and purify by silica gel column chromatography to obtain the product.

[0014] The present invention has found through experiments that compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one can significantly inhibit H 2 O 2 Mediates oxidative stress-induced loss of PC12 cells and increases cell viability, and resists H 2 O 2 The stronger the ability of oxidative stress. Compared with the control group, the compound at a concentration of 10 μM restored 90% of the cell activity, with no significant difference.

[0015] Therefore, the present invention provides the use of the compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one in the preparation of a drug for inhibiting oxidative stress damage of nerve cells.

[0016] Since oxidative stress-induced nerve damage is the main pathological mechanism of neurodegenerative diseases, all drugs that can inhibit oxidative stress damage are likely to be potential drugs for treating neurodegenerative diseases. On PC12 neural cells, the compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one has an antioxidant effect at low concentrations, so the present invention also provides the use of the compound in the preparation of drugs for treating neurodegenerative diseases.

[0017] Beneficial effects of the present invention:

[0018] Due to the complexity of brain function, the treatment of neurodegenerative diseases has always been a difficult problem in medicine. The pathology of neurodegenerative diseases involves many molecular mechanisms, such as oxidative stress, mitochondrial damage, excitotoxicity, etc. The present invention uses the oxidative stress model of nerve cells to study the neuroprotective effect of compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one, which is of great significance for the treatment of neurodegenerative diseases. Therefore, the technical solution of the present invention provides a new choice for the research and development of drugs for the treatment of neurodegenerative diseases. The drug capable of inhibiting oxidative stress damage provided by the present invention is a new use of an old drug and can be used as a potential drug for the treatment of neurodegenerative diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The compound of the present invention (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one 1 H-NMR spectrum;

[0020] Figure 2 The compound of the present invention (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one 13 C-NMR spectrum;

[0021] Figure 3 is the HRMS spectrum of the compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one of the present invention;

[0022] Figure 4 The results of CCK-8 detection of cell activity in the present invention; ***P<0.001, NS indicates no significant difference. DETAILED DESCRIPTION

[0023] The present invention is further described in detail below with specific embodiments, but it should not be understood that the scope of the above subject matter of the present invention is limited to the following embodiments, and all technologies realized based on the content of the present invention belong to the scope of the present invention.

[0024] Unless otherwise specified, all reagents and materials used in the present invention are commercially available.

[0025] Example 1

[0026] The preparation method of compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one comprises the following steps:

[0027]

[0028] 116.64 mg of 3,5-dimethoxyacetophenone was added to a 10 mL round-bottom flask, 1 mL of anhydrous ethanol was added to dissolve, 0.5 mL of 60% sodium hydroxide aqueous solution was added, and the mixture was stirred at room temperature for 10 min. 74.5 mg of 3,4-dihydroxybenzaldehyde was added, and the mixture was stirred at 85 ° C for 15 min. The mixture was cooled to room temperature, acidified with hydrochloric acid, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-1:10, v / v) to obtain 57.5 mg of the product with a yield of 35.5%.

[0029] like Figure 1 As shown, 1 H-NMR (400 MHz, DMSO-d 6 )δ7.36(d,J=15.8Hz,1H),7.15–7.06(m,4H),7.01(dd,J=6.8,2.4Hz,2H),6.78(d,J=8.1Hz,1H),3.81(s,3H),3.74(s,3H). like Figure 2 As shown, 13 C-NMR (101 MHz, DMSO-d 6 )δ191.49,153.06,151.69,148.74,145.69,143.92,129.78,126.03,123.36,122.10,118.06,115.89,114.68,113.94,113.90,56.38,55.55. Figure 3 As shown, HRMS (ESI) calculated for C 17 H 16 O 5 [M+H] + :301.1071, found 301.1075.

[0030] Example 2

[0031] The purpose of this example is to study the protective effect of compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one on oxidative stress injury of PC12 cells.

[0032] 1. Experimental Methods

[0033] 1.1 PC12 cell culture

[0034] PC12 cells purchased from Wuhan ATCC Cell Bank were cultured in DMEM containing 10% FBS, 100 U / mL penicillin and 100 U / mL streptomycin at 37°C, 95% air and 5% CO 2The cells were cultured in a cell culture incubator at 1×10 5 The cells were plated in 96-well plates at a density of 100 cells / well. After 24 hours, PC12 cells were first treated with 300 μM H 2 O 2 The cells were treated for 30 minutes. Then, the compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one was added to give a final concentration of 1, 5, 10, 20 and 40 μM. After 24 hours, the cell activity was detected using the Cell Counting Kit-8.

[0035] 1.2CCK-8 detection

[0036] According to the instructions of the reagent, the cell viability was detected using the CCK-8 kit. After the PC12 cells seeded in the 96-well plate were treated with the compound for 24 hours, 10 μL of CCK-8 solution was added to each well and incubated at 37°C for 30 minutes. Thereafter, the absorbance at 450 nm was measured using a microplate reader (BioTek).

[0037] The cell viability calculation formula is as follows:

[0038]

[0039] 2. Experimental results

[0040] The compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one significantly inhibited H 2 O 2 Mediates oxidative stress-induced PC12 cell loss and enhances cell viability.

[0041] In order to further study the in vivo effects of the compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one, H 2 O 2 Induce oxidative stress damage in PC12 cells. PC12 cells were first added with 300 μM H 2 O 2 After incubation for 30 minutes, the cells were treated with different final concentrations (1, 5, 10, 20 and 40 μM) of compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one and incubated for 24 hours. Cell activity was detected by CCK-8. Figure 4 As shown, compared with the control group, 300 μM H 2 O 2 The induced cell count decreased by 55% and the cell activity was significantly reduced (P<0.001). 2O 2 Compared with the control group, 1μM and 5μM concentrations of compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one significantly blocked the H 2 O 2 The cell survival rate induced by H was decreased (P<0.001), and the resistance to H 2 O 2 The stronger the ability to resist oxidative stress. Compared with the control group, the 10μM compound restored 90% of the cell activity, with no significant difference. After that, as the compound concentration (20, 40μM) increased, the biological activity of the compound decreased. The 40μM compound no longer had significant antioxidant activity.

[0042] Since oxidative stress-induced nerve damage is the main pathological mechanism of neurodegenerative diseases, all drugs that can inhibit oxidative stress damage are likely to be potential drugs for the treatment of neurodegenerative diseases. Studies have found that the synthetic compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one does have an antioxidant effect at low concentrations on PC12 neural cells, so it has the potential to treat neurodegenerative diseases.

Claims

1. A method for preparing an alkoxy-substituted chalcone derivative, characterized in that: The alkoxy-substituted chalcone derivative refers to the compound (E)-3-(3,4-dihydroxyphenyl)-1-(2,5-dimethoxyphenyl)prop-2-en-1-one, with the structural formula: The method comprises the following steps: adding 3,5-dimethoxyacetophenone into a reaction container, adding anhydrous ethanol to dissolve, adding a 60% sodium hydroxide aqueous solution, stirring at room temperature, adding 3,4-dihydroxybenzaldehyde, stirring at 70-90°C for reaction, cooling to room temperature after the reaction is completed, acidifying with hydrochloric acid, extracting with ethyl acetate, drying with anhydrous sodium sulfate, concentrating under reduced pressure, and purifying through silica gel column chromatography to obtain a product.

2. Use of the alkoxy-substituted chalcone derivative according to claim 1 in the preparation of a drug for inhibiting oxidative stress damage of nerve cells.

3. Use of the alkoxy-substituted chalcone derivative according to claim 1 in the preparation of a drug for treating neurodegenerative diseases.

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