A benzopyran chalcone compound, preparation method and application and pharmaceutical preparation thereof
By synthesizing benzopyran chalcone compounds and applying them to various pharmaceutical formulations, the problem of poor efficacy of existing anti-inflammatory drugs has been solved, enabling effective treatment of inflammatory diseases such as acute lung injury, while ensuring biocompatibility.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- WENZHOU MEDICAL UNIV
- Filing Date
- 2023-11-20
- Publication Date
- 2026-06-19
AI Technical Summary
Existing anti-inflammatory drugs are not very effective and have side effects, making it difficult to effectively control inflammatory diseases such as acute lung injury, inflammatory bowel disease, skin inflammation, asthma, and joint pain. Furthermore, long-term use increases medical costs and delays the treatment process.
Benzopyran chalcone compounds were designed and synthesized, prepared through a specific chemical reaction route, and applied to anti-inflammatory drugs, including forms such as injections, tablets, capsules, aerosols, suppositories, films, pellets, ointments, controlled-release agents, and nano-formulations.
Benzopyran chalcone compounds exhibit significant anti-inflammatory activity, particularly in the treatment of acute lung injury, and have high biocompatibility, without causing significant hepatotoxicity or nephrotoxicity, providing a more stable treatment option.
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Figure CN117551085B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of pharmaceutical chemistry, and in particular to a benzopyran chalcone compound, its preparation method, its application, and its pharmaceutical formulation. Background Technology
[0002] Inflammatory diseases are a widespread medical problem globally, encompassing many different conditions, including but not limited to acute lung injury, inflammatory bowel disease, skin inflammation, asthma, and joint pain. These diseases share a common characteristic: the excessive secretion of inflammatory cytokines, causing severe inflammatory responses that lead to chronic pain, tissue damage, and disease. These diseases result in a significant decline in the quality of life for many patients, and can even cause severe disability and death.
[0003] Currently, medications for treating inflammatory diseases mainly include nonsteroidal anti-inflammatory drugs (NSAIDs), steroids, and immunosuppressants. While these methods can control inflammation to some extent, they often come with a range of problems. For example, long-term use of NSAIDs can cause gastrointestinal ulcers and bleeding, steroids can lead to immune system suppression and other side effects, and immunosuppressants may increase the risk of infection. Furthermore, the efficacy of existing medications is inconsistent, and patients may need to try multiple drugs to find the most suitable treatment. This not only increases medical costs but also delays disease management and recovery for patients.
[0004] Therefore, it is of great significance to study a benzopyran chalcone compound, its preparation method, and to formulate it into an anti-inflammatory drug. Summary of the Invention
[0005] The purpose of this invention is to provide a benzopyran chalcone compound, its preparation method, its application, and its pharmaceutical formulation, in order to solve the problem of poor therapeutic effects of existing anti-inflammatory drugs.
[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:
[0007] This invention provides a benzopyran chalcone compound having the structure shown in formula (I):
[0008]
[0009] This invention also provides a method for preparing the above-mentioned benzopyran chalcone compounds. The synthetic route for benzopyran chalcone compounds is as follows:
[0010]
[0011] The preparation method of benzopyran chalcone compounds includes the following steps:
[0012] (1) Compound 1, potassium carbonate, and chloromethyl methyl ether were mixed in acetone and reacted to obtain compound 2;
[0013] (2) Compound 2, sodium hydroxide, tetrabutylammonium bromide and dimethyl sulfate were mixed in a solvent and reacted to obtain compound 3;
[0014] (3) Compound 3, anhydrous ethanol and hydrochloric acid were mixed and reacted to obtain compound 4;
[0015] (4) Compound 4, ethylenediamine diacetate solution and 3-methyl-2-butenal were mixed and reacted to obtain compound 5;
[0016] (5) Compounds 5 and 6 were mixed in anhydrous ethanol and reacted to obtain benzopyran chalcone compounds.
[0017] Preferably, in step (1), the molar ratio of compound 1, potassium carbonate and chloromethyl methyl ether is 0.5-1.5:2-5:2-3; the molar volume ratio of compound 1 and acetone is 0.5-1.5 mmol:20-80 mL; the reaction temperature is 50-70 °C; and the reaction time is 2-4 h.
[0018] Preferably, in step (2), the molar ratio of compound 2, sodium hydroxide, tetrabutylammonium bromide, and dimethyl sulfate is 1:1-2:0.05-0.2:2.0-3.0; the molar volume ratio of compound 2 to solvent is 1 mmol:30-50 mL, wherein the solvent includes dichloromethane and water; the reaction temperature is 20-30°C, and the reaction time is 1-3 h.
[0019] Preferably, in step (3), the molar ratio of compound 3 to HCl in hydrochloric acid is 1:0.5 to 1.5; the molar volume ratio of compound 3 to anhydrous ethanol is 1 mmol:30 to 50 mL; the reaction temperature is 40 to 60 °C, and the reaction time is 2 to 4 h.
[0020] Preferably, in step (4), the molar ratio of compound 4, 3-methyl-2-butenal, and ethylenediamine diacetate is 0.5–0.6:0.6–0.7:0.02–0.07; the concentration of the ethylenediamine diacetate solution is 0.005–0.02 mmol / mL; the reaction temperature is 110–130 °C, and the reaction time is 8–12 h.
[0021] Preferably, in step (5), the molar ratio of compound 5 to compound 6 is 1:1 to 1.5; the molar volume ratio of compound 5 to anhydrous ethanol is 1 mmol: 50 to 80 mL; the reaction temperature is 90 to 110 °C; and the reaction time is 45 to 55 h.
[0022] The present invention also provides the application of the above-mentioned benzopyran chalcone compounds in the preparation of anti-inflammatory drugs.
[0023] Preferably, the inflammation includes acute lung injury, intrauterine adhesions, inflammatory bowel disease, skin inflammation, asthma, or joint pain.
[0024] The present invention also provides a pharmaceutical preparation comprising an active ingredient and excipients, wherein the active ingredient comprises the above-mentioned benzopyran chalcone compounds; the pharmaceutical preparation is an injection, tablet, capsule, aerosol, suppository, film, drop pill, ointment, controlled-release agent, sustained-release agent or nano-formulation.
[0025] The beneficial effects of this invention are:
[0026] Based on the benzopyran structural skeleton, a benzopyran chalcone compound (I) was designed and synthesized using drug design methods. The benzopyran chalcone compound (I) of this invention exhibits excellent anti-inflammatory activity, especially showing significant therapeutic effects on acute lung injury. Attached Figure Description
[0027] Figure 1 The benzopyran chalcone compound (I) prepared in Example 1 1 H NMR spectrum;
[0028] Figure 2 The benzopyran chalcone compound (I) prepared in Example 1 13 C10 NMR spectrum;
[0029] Figure 3For example 1, the in vitro anti-inflammatory activity (IL-6 and TNF-α) of the benzopyran chalcone compound (I) prepared in Example 1 was tested. In this diagram, A shows the transcriptional level of TNF-α inflammatory factor in the control group, LPS model group, and benzopyran chalcone compound (I) treatment group; B shows the transcriptional level of IL-6 inflammatory factor in the control group, LPS model group, and benzopyran chalcone compound (I) treatment group; C shows the protein expression level of TNF-α inflammatory factor in the control group, LPS model group, and benzopyran chalcone compound (I) treatment group; and D shows the protein expression level of IL-6 inflammatory factor in the control group, LPS model group, and benzopyran chalcone compound (I) treatment group.
[0030] Figure 4 For Test Example 2, the in vivo anti-inflammatory activity of the benzopyran chalcone compound (I) prepared in Example 1 was tested. In the figure, A is the content of TNF-α inflammatory factor in the serum of mice in the control group, LPS model group and benzopyran chalcone compound (I) treatment group; B is the content of TNF-α inflammatory factor in the bronchoalveolar lavage fluid of mice in the control group, LPS model group and benzopyran chalcone compound (I) treatment group; C is the content of IL-6 inflammatory factor in the serum of mice in the control group, LPS model group and benzopyran chalcone compound (I) treatment group; D is the content of IL-6 inflammatory factor in the bronchoalveolar lavage fluid of mice in the control group, LPS model group and benzopyran chalcone compound (I) treatment group; E is the morphological structure of the lung tissue of mice in the control group, LPS model group and benzopyran chalcone compound (I) treatment group.
[0031] Figure 5 The graph shows the results of a biosafety assessment of the benzopyran chalcone compound (I) prepared in Example 1 for Test Example 3. In Test Example 3, A represents the ALT content in the blood of mice at the control group, therapeutic concentration of 50 mg / kg, and therapeutic concentration of 100 mg / kg; B represents the AST content in the blood of mice at the control group, therapeutic concentration of 50 mg / kg, and therapeutic concentration of 100 mg / kg; and C represents the Cr content in the blood of mice at the control group, therapeutic concentration of 50 mg / kg, and therapeutic concentration of 100 mg / kg. Detailed Implementation
[0032] This invention provides a benzopyran chalcone compound having the structure shown in formula (I):
[0033]
[0034] This invention also provides a method for preparing the above-mentioned benzopyran chalcone compounds. The synthetic route for benzopyran chalcone compounds is as follows:
[0035]
[0036] The preparation method of benzopyran chalcone compounds includes the following steps:
[0037] (1) Compound 1, potassium carbonate, and chloromethyl methyl ether were mixed in acetone and reacted to obtain compound 2;
[0038] (2) Compound 2, sodium hydroxide, tetrabutylammonium bromide and dimethyl sulfate were mixed in a solvent and reacted to obtain compound 3;
[0039] (3) Compound 3, anhydrous ethanol and hydrochloric acid were mixed and reacted to obtain compound 4;
[0040] (4) Compound 4, ethylenediamine diacetate solution and 3-methyl-2-butenal were mixed and reacted to obtain compound 5;
[0041] (5) Compounds 5 and 6 were mixed in anhydrous ethanol and reacted to obtain benzopyran chalcone compounds.
[0042] In this invention, in step (1), the molar ratio of compound 1, potassium carbonate, and chloromethyl methyl ether is 0.5–1.5:2–5:2–3, preferably 0.8–1.2:3–4:2.2–2.8, and more preferably 1.0:4:2.5; the molar volume ratio of compound 1 and acetone is 0.5–1.5 mmol:20–80 mL, preferably 0.8–1.2 mmol:30–70 mL, and more preferably 1.0 mmol:40–60 mL; the reaction temperature is 50–70 °C, preferably 55–65 °C, and more preferably 60 °C; and the reaction time is 2–4 h, preferably 2.5–3.5 h, and more preferably 3 h.
[0043] In this invention, in step (2), the molar ratio of compound 2, sodium hydroxide, tetrabutylammonium bromide, and dimethyl sulfate is 1:1-2:0.05-0.2:2.0-3.0, preferably 1:1.2-1.8:0.08-0.15:2.1-2.5, and more preferably 1:1.5:0.1-0.12:2.2-2.3; the molar volume ratio of compound 2 to solvent is 1 mmol:30-50 mL, preferably 1 mmol:35-45 mL, and more preferably 1 mmol:40 mL, and the solvent comprises dichloromethane and water; the volume ratio of dichloromethane to water is 3:2; the reaction temperature is 20-30°C, preferably 22-28°C, and more preferably 25°C; the reaction time is 1-3 h, preferably 1.5-2.5 h, and more preferably 2 h.
[0044] In this invention, in step (3), the molar ratio of compound 3 to HCl in hydrochloric acid is 1:0.5-1.5, preferably 1:0.8-1.2, and more preferably 1:1.0; the molar volume ratio of compound 3 to anhydrous ethanol is 1 mmol:30-50 mL, preferably 1 mmol:35-45 mL, and more preferably 1 mmol:40 mL; the reaction temperature is 40-60°C, preferably 45-55°C, and more preferably 50°C; the reaction time is 2-4 h, preferably 2.5-3.5 h, and more preferably 3 h.
[0045] In this invention, in step (4), the molar ratio of compound 4, 3-methyl-2-butenal, and ethylenediamine diacetate is 0.5–0.6:0.6–0.7:0.02–0.07, preferably 0.52–0.58:0.62–0.68:0.04–0.06, and more preferably 0.55:0.64–0.66:0.05; the ethylenediamine diacetate solution is preferably prepared from ethylenediamine diacetate and xylene, and the concentration of the ethylenediamine diacetate solution is 0.005–0.02 mmol / mL, preferably 0.008–0.015 mmol / mL, and more preferably 0.01–0.012 mmol / mL; the reaction temperature is 110–130°C, preferably 115–125°C, and more preferably 120°C; the reaction time is 8–12 h, preferably 9–11 h, and more preferably 10 h.
[0046] In this invention, in step (5), the molar ratio of compound 5 to compound 6 is 1:1 to 1.5, preferably 1:1.1 to 1.4, and more preferably 1:1.2 to 1.3; the molar volume ratio of compound 5 to anhydrous ethanol is 1 mmol: 50 to 80 mL, preferably 1 mmol: 55 to 70 mL, and more preferably 1 mmol: 60 mL; the reaction temperature is 90 to 110 °C, preferably 95 to 105 °C, and more preferably 100 °C; the reaction time is 45 to 55 h, preferably 46 to 52 h, and more preferably 48 to 50 h.
[0047] The present invention also provides the application of the above-mentioned benzopyran chalcone compounds in the preparation of anti-inflammatory drugs.
[0048] In this invention, the inflammation includes acute lung injury, intrauterine adhesions, inflammatory bowel disease, skin inflammation, asthma, or joint pain.
[0049] The present invention also provides a pharmaceutical preparation comprising an active ingredient and pharmaceutical excipients, wherein the active ingredient comprises the above-mentioned benzopyran chalcone compounds; the pharmaceutical preparation is an injection, tablet, capsule, aerosol, suppository, film, drop pill, ointment, controlled-release agent, sustained-release agent or nano-formulation.
[0050] In this invention, the pharmaceutical preparation is preferably an injection, tablet, capsule, aerosol, suppository, sustained-release preparation or nano-preparation, and more preferably an injection, tablet or capsule.
[0051] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.
[0052] Compound 1 in the embodiments of the present invention is Compound 6 is
[0053] Example 1
[0054] 1.00 mmol of compound 1 and 4.00 mmol of potassium carbonate were mixed in 40 mL of acetone and heated to 60 °C. Then, 2.5 mmol of chloromethyl methyl ether was added dropwise to the reaction mixture over 30 min. The mixture was refluxed for 3 h. After the reaction was completed, the mixture was cooled to room temperature. Finally, the reaction mixture was filtered and the solvent was removed under reduced pressure. The product was then subjected to column chromatography to give a white oily product (compound 2) with a yield of 55%.
[0055] 1.00 mmol of compound 2 was dissolved in 40 mL of a mixed solvent of dichloromethane / deionized water (the volume ratio of dichloromethane to deionized water was 3:2). Then, 1.50 mmol of sodium hydroxide and 0.10 mmol of tetrabutylammonium bromide were added, followed by the dropwise addition of 2.20 mmol of dimethyl sulfate. The mixture was stirred at 12000 rpm at 25 °C for 2 h. The resulting mixture was heated at 60 °C for 30 min to destroy the remaining dimethyl sulfate. The mixture was then extracted with dichloromethane. The organic layer was dried with anhydrous magnesium sulfate. The solvent was removed under reduced pressure at 4.5 kPa at room temperature. The product was purified by silica gel column chromatography to obtain a white oily product (compound 3) with a yield of 80%.
[0056] 1.00 mmol of compound 3 was dissolved in 40 mL of anhydrous ethanol, and 3 M hydrochloric acid (containing 1 mmol of HCl) was added. The mixture was then heated to 50 °C and reacted for 3 h. After cooling to room temperature, the mixture was poured into deionized water and extracted three times with ethyl acetate. The combined organic layers were dried with anhydrous magnesium sulfate and concentrated under vacuum at 9.3 kPa at room temperature. The product was purified by silica gel column chromatography to obtain a white solid product (compound 4) with a yield of 85%.
[0057] 0.55 mmol of compound 4, 0.64 mmol of 3-methyl-2-butenal, and 5 mL of ethylenediamine diacetate solution (the ethylenediamine diacetate solution was prepared by ethylenediamine diacetate and xylene, and the concentration of the ethylenediamine diacetate solution was 0.01 mmol / mL) were mixed and reacted at 120 °C for 10 h. After cooling to room temperature, the mixture was poured into deionized water and extracted with ethyl acetate. The combined organic layers were dried with anhydrous magnesium sulfate and concentrated under vacuum. The product was purified by silica gel column chromatography to obtain a yellow solid product (compound 5) with a yield of 65%.
[0058] 1.00 mmol of compound 5 and 1.2 mmol of compound 6 were mixed in 60 mL of anhydrous ethanol and reacted at 95 °C for 48 h. After the reaction was completed, the reaction was quenched with 25% hydrochloric acid to pH 6, and then extracted with ethyl acetate. The organic layer was dried with anhydrous magnesium sulfate and concentrated under vacuum. The product was purified by silica gel column chromatography to obtain a yellow solid product, which is benzopyran chalcone compound (I), with a yield of 45%.
[0059] Structural characterization data:
[0060] 1H NMR (400MHz, DMSO-d6) δ14.72(s,1H),11.39(s,1H),7.91(d,J=9.7Hz,2H),7.85(d,J=15.5Hz,1H),7 .55–7.40(m,3H),6.59–6.51(m,2H),6.12(s,1H),5.60(d,J=10.0Hz,1H),3.95(s,3H),1.42(s,6H).
[0061] 13 C NMR(101MHz,DMSO-d6)δ192.64,162.92,161.81,160.15,146.22,137.98,128.57,127.30,126.4 9,123.74,123.62,121.50,115.81,112.80,105.99,102.79,102.54,92.42,78.57,56.90,28.52.
[0062] Example 2
[0063] 0.50 mmol of compound 1 and 2.0 mmol of potassium carbonate were mixed in 20 mL of acetone and heated to 50 °C. Then, 2.0 mmol of chloromethyl methyl ether was added dropwise to the reaction mixture over 30 min. The mixture was refluxed for 4 h. After the reaction was completed, the mixture was cooled to room temperature. Finally, the reaction mixture was filtered and the solvent was removed under reduced pressure. The product was then subjected to column chromatography to obtain a white oily product (compound 2) with a yield of 52%.
[0064] 1.00 mmol of compound 2 was dissolved in 30 mL of a mixed solvent of dichloromethane / deionized water (the volume ratio of dichloromethane to deionized water was 3:2). Then, 1.0 mmol of sodium hydroxide and 0.05 mmol of tetrabutylammonium bromide were added, followed by the dropwise addition of 2.0 mmol of dimethyl sulfate. The mixture was stirred at 12000 rpm at 20 °C for 3 h. The resulting mixture was heated at 60 °C for 30 min to destroy the remaining dimethyl sulfate. The mixture was then extracted with dichloromethane. The organic layer was dried with anhydrous magnesium sulfate. The solvent was removed under reduced pressure at 3.3 kPa at room temperature. The product was purified by silica gel column chromatography to obtain a white oily product (compound 3) with a yield of 75%.
[0065] 1.00 mmol of compound 3 was dissolved in 30 mL of anhydrous ethanol, and 3 M hydrochloric acid (containing 0.5 mmol of HCl) was added. The mixture was then heated to 60 °C and reacted for 2 h. After cooling to room temperature, the mixture was poured into deionized water and extracted three times with ethyl acetate. The combined organic layers were dried with anhydrous magnesium sulfate and concentrated under vacuum. The product was purified by silica gel column chromatography to obtain a white solid product (compound 4). The yield of compound 4 was 81%.
[0066] 0.6 mmol of compound 4, 0.7 mmol of 3-methyl-2-butenal, and 10 mL of ethylenediamine diacetate solution (the ethylenediamine diacetate solution was prepared by ethylenediamine diacetate and xylene, and the concentration of the ethylenediamine diacetate solution was 0.006 mmol / mL) were reacted at 110 °C for 12 h. After cooling to room temperature, the mixture was poured into deionized water and extracted with ethyl acetate. The combined organic layers were dried with anhydrous magnesium sulfate and concentrated under vacuum at 6.6 kPa at room temperature. The product was purified by silica gel column chromatography to obtain a yellow solid product (compound 5) with a yield of 67%.
[0067] 1.00 mmol of compound 5 and 1 mmol of compound 6 were mixed in 80 mL of anhydrous ethanol and reacted at 90 °C for 55 h. After the reaction was completed, the reaction was quenched with 25% hydrochloric acid to pH 6, and then extracted with ethyl acetate. The organic layer was dried with anhydrous magnesium sulfate and concentrated under vacuum. The product was purified by silica gel column chromatography to obtain a yellow solid product, which is benzopyran chalcone compound (I), with a yield of 43%.
[0068] Example 3
[0069] 1.50 mmol of compound 1 and 5.0 mmol of potassium carbonate were mixed in 60 mL of acetone and heated to 70 °C. Then, 3.0 mmol of chloromethyl methyl ether was added dropwise to the reaction mixture over 30 min. The mixture was refluxed for 2 h. After the reaction was completed, the mixture was cooled to room temperature. Finally, the reaction mixture was filtered and the solvent was removed under reduced pressure. The product was then subjected to column chromatography to give a white oily product (compound 2) with a yield of 58%.
[0070] 1.00 mmol of compound 2 was dissolved in 50 mL of a mixed solvent of dichloromethane / deionized water (the volume ratio of dichloromethane to deionized water was 3:2). Then, 2.0 mmol of sodium hydroxide and 0.2 mol of tetrabutylammonium bromide were added, followed by the dropwise addition of 3.0 mmol of dimethyl sulfate. The mixture was stirred at 12000 rpm at 30 °C for 1 h. The resulting mixture was heated at 60 °C for 30 min to destroy the remaining dimethyl sulfate. The mixture was then extracted with dichloromethane. The organic layer was dried with anhydrous magnesium sulfate. The solvent was removed under reduced pressure at 6.6 kPa at room temperature. The product was purified by silica gel column chromatography to obtain a white oily product (compound 3) with a yield of 83%.
[0071] 1.00 mmol of compound 3 was dissolved in 50 mL of anhydrous ethanol, and 3 M hydrochloric acid (containing 1.5 mmol of HCl) was added. The mixture was then heated to 40 °C and reacted for 4 h. After cooling to room temperature, the mixture was poured into deionized water and extracted three times with ethyl acetate. The combined organic layers were dried with anhydrous magnesium sulfate and concentrated under vacuum. The product was purified by silica gel column chromatography to obtain a white solid product (compound 4) with a yield of 88%.
[0072] 0.5 mmol of compound 4, 0.6 mmol of 3-methyl-2-butenal, and 4 mL of ethylenediamine diacetate solution (the ethylenediamine diacetate solution was prepared by ethylenediamine diacetate and xylene, and the concentration of the ethylenediamine diacetate solution was 0.01 mmol / mL) were reacted at 130 °C for 8 h. After cooling to room temperature, the mixture was poured into deionized water and extracted with ethyl acetate. The combined organic layers were dried with anhydrous magnesium sulfate and concentrated under vacuum at 8.5 kPa at room temperature. The product was purified by silica gel column chromatography to obtain a yellow solid product (compound 5) with a yield of 70%.
[0073] 1.00 mmol of compound 5 and 1.5 mmol of compound 6 were mixed in 50 mL of anhydrous ethanol and reacted at 110 °C for 45 h. After the reaction was completed, the reaction was quenched with 25% hydrochloric acid to pH 6, and then extracted with ethyl acetate. The organic layer was dried with anhydrous magnesium sulfate and concentrated under vacuum. The product was purified by silica gel column chromatography to obtain a yellow solid product, which is benzopyran chalcone compound (I), with a yield of 48%.
[0074] Test Example 1
[0075] In vitro anti-inflammatory activity (IL-6 and TNF-α) of benzopyran chalcone compounds: A classic in vitro inflammatory cell model was constructed using LPS-stimulated Raw264.7 macrophages to detect the anti-inflammatory activity of the synthesized benzopyran chalcone compounds. Although only the LPS-stimulated Raw264.7 inflammatory cell model is used as an example to illustrate its use in preparing anti-inflammatory drugs, the term "anti-inflammatory" in this invention includes, but is not limited to, LPS-induced inflammatory responses in Raw264.7 cells. In this test example, the Raw264.7 cell line was used for preliminary in vitro anti-inflammatory activity testing. The cells were stimulated to secrete inflammatory factors TNF-α and IL-6 by adding 0.5 μg / mL of LPS. The effects of a 10 μM drug concentration on the transcriptional levels of TNF-α and IL-6 inflammatory factors at 6 h and on the protein expression levels of TNF-α and IL-6 inflammatory factors at 24 h were tested.
[0076] Experimental results are as follows Figure 3 China A and Figure 3 As shown in Figure B, LPS stimulation significantly increased the transcriptional levels of TNF-α and IL-6 inflammatory factors in Raw264.7 cells; while treatment with 10 μM benzopyran chalcone compound (I) significantly reduced the transcriptional levels of TNF-α and IL-6 inflammatory factors. Figure 3 C and Figure 3 The experimental results from D further showed that treatment with 10 μM benzopyran chalcone compound (I) significantly reduced the protein expression levels of LPS-induced TNF-α and IL-6 inflammatory factors.
[0077] Test Example 2
[0078] Tests on the in vivo anti-inflammatory activity of benzopyran chalcone compounds (alleviation of LPS-induced acute lung injury): BALB / c (C57 black mice) were acclimatized for one week, and an acute lung injury model was established by intratracheal instillation of LPS (LPS dosage in mice was 5 mg / kg, dissolved in physiological saline). To further verify the in vivo anti-inflammatory activity of benzopyran chalcone compound (I), the concentrations of TNF-α and IL-6 in the bronchoalveolar lavage fluid (BALF) and blood of mice were measured using ELISA. The experimental results are as follows: Figure 4 As shown in Figures A through D, benzopyran chalcone compounds (I) effectively inhibited the release of inflammatory factors TNF-α and IL-6 in both blood and lavage fluid.
[0079] The effects of benzopyran chalcone compound (I) on LPS-induced acute lung injury were tested. Further hematoxylin and eosin (H&E) staining experiments were used to observe the lung tissue morphology of mice in the control group, LPS-induced model group, and benzopyran chalcone compound (I) treatment group. The experimental results are as follows: Figure 4 As shown in Figure E, compared with the control group, the lung tissue of mice in the LPS model group showed significant thickening of alveolar walls, formation of hyaline membranes, significant damage to alveolar structure, and alveolar collapse. The alveolar tissue of mice in the benzopyran chalcone compound (I) treatment group was similar to that of the control group, indicating that benzopyran chalcone compound (I) can effectively alleviate LPS-induced acute lung injury.
[0080] Test Example 3
[0081] ALT (alanine aminotransferase) and AST (aspartate aminotransferase) are two commonly used enzyme indicators in liver function tests to assess the health of liver function. Under normal circumstances, the levels of these two enzymes are low. When the liver is damaged or diseased, they are usually released into the bloodstream, causing their levels to rise. Cr (creatinine) is a muscle metabolic product that is excreted through the kidneys. The kidneys are the body's main excretory organs, responsible for removing waste and metabolic products from the blood. When kidney function is impaired, the body's ability to clear creatinine decreases, leading to elevated creatinine levels in the blood. Therefore, blood creatinine levels are commonly used as an indicator of kidney function. Elevated creatinine levels may be a sign of kidney dysfunction. Therefore, in order to further evaluate the biosafety of benzopyran chalcone compound (I), mice were treated with 5 times (50 mg / kg) and 10 times (100 mg / kg) concentrations of benzopyran chalcone compound (I), and blood was drawn 7 days later to detect the levels of ALT, AST and Cr.
[0082] Experimental results are as follows Figure 5 As shown, continuous administration of 50 mg / kg and 100 mg / kg of benzopyran chalcone compound (I) for 7 days did not cause significant changes in the levels of ALT, AST and Cr in the blood of mice, indicating that it has high biocompatibility.
[0083] As can be seen from the above embodiments, the present invention provides a benzopyran chalcone compound, its preparation method, and its application. Based on the benzopyran structural skeleton, the present invention designs and synthesizes a benzopyran chalcone compound (I) using drug design methods. The benzopyran chalcone compound of the present invention exhibits excellent anti-inflammatory activity and high biocompatibility.
[0084] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. A benzopyran chalcone compound, characterized in that, The benzopyran chalcone compounds have the structure shown in formula (I): （I）。 2. The method for preparing the benzopyran chalcone compound according to claim 1, characterized in that, The synthetic routes for benzopyran chalcone compounds are as follows: The preparation method of benzopyran chalcone compounds includes the following steps: (1) Compound 1, potassium carbonate, and chloromethyl methyl ether were mixed in acetone and reacted to obtain compound 2; (2) Compound 2, sodium hydroxide, tetrabutylammonium bromide and dimethyl sulfate were mixed in a solvent and reacted to obtain compound 3; (3) Compound 3, anhydrous ethanol and hydrochloric acid were mixed and reacted to obtain compound 4; (4) Compound 4, ethylenediamine diacetate solution and 3-methyl-2-butenal were mixed and reacted to obtain compound 5; (5) Compound 5 and Compound 6 were mixed in anhydrous ethanol and reacted to obtain benzopyran chalcone compounds.
3. The preparation method according to claim 2, characterized in that, In step (1), the molar ratio of compound 1, potassium carbonate and chloromethyl methyl ether is 0.5~1.5:2~5:2~3; the molar volume ratio of compound 1 and acetone is 0.5~1.5mmol:20~80mL; the reaction temperature is 50~70℃ and the reaction time is 2~4h.
4. The preparation method according to claim 2 or 3, characterized in that, In step (2), the molar ratio of compound 2, sodium hydroxide, tetrabutylammonium bromide, and dimethyl sulfate is 1:1~2:0.05~0.2:2.0~3.0; the molar volume ratio of compound 2 to solvent is 1 mmol:30~50 mL, the solvent is a mixture of dichloromethane and deionized water, and the volume ratio of dichloromethane to water is 3:2; the reaction temperature is 20~30℃, and the reaction time is 1~3 h.
5. The preparation method according to claim 4, characterized in that, In step (3), the molar ratio of compound 3 to HCl in hydrochloric acid is 1:0.5~1.5; the molar volume ratio of compound 3 to anhydrous ethanol is 1 mmol:30~50 mL; the reaction temperature is 40~60℃, and the reaction time is 2~4 h.
6. The preparation method according to claim 3 or 5, characterized in that, In step (4), The molar ratio of compound 4, 3-methyl-2-butenal, and ethylenediamine diacetate is 0.5~0.6:0.6~0.7:0.02~0.07; the concentration of the ethylenediamine diacetate solution is 0.005~0.02 mmol / mL; the reaction temperature is 110~130℃, and the reaction time is 8~12 h.
7. The preparation method according to claim 6, characterized in that, In step (5), the molar ratio of compound 5 to compound 6 is 1:1~1.5; the molar volume ratio of compound 5 to anhydrous ethanol is 1 mmol: 50~80 mL; the reaction temperature is 90~110℃, and the reaction time is 45~55 h.
8. The use of the benzopyran chalcone compound of claim 1 in the preparation of an anti-acute lung injury inflammatory drug.
9. A pharmaceutical preparation, characterized in that, The pharmaceutical preparation includes an active ingredient and excipients, wherein the active ingredient includes the benzopyran chalcone compounds as described in claim 1; the pharmaceutical preparation is an injection, tablet, capsule, aerosol, suppository, film, drop pill, ointment, controlled-release agent, sustained-release agent, or nano-formulation.