A benzene-alpha-pyrone monocarbonyl curcuminoid and its preparation method and application

Abstract

The application provides a benzene-alpha-pyranyl monocarbonyl curcumin analogue as well as a preparation method and application thereof, and belongs to the technical field of medicinal chemistry. The benzene-alpha-pyranyl monocarbonyl curcumin analogue with the structure of formula (I) is synthesized based on a curcumin skeleton and combined with a pharmacological pharmacological group of xanthohumol compounds, has excellent antitumor activity on cervical cancer, liver cancer, osteosarcoma and ovarian cancer, and has low toxic and side effects on normal liver cells.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, and in particular to a benzo-α-pyran-type monocarbonyl curcumin analog, its preparation method, and its application. Background Technology

[0002] Cancer has become a disease with high incidence and mortality rates, seriously threatening human health. In recent decades, with the rapid development of medical techniques, breakthroughs have been made in cancer treatment. However, the treatment of most cancers in clinical practice still mainly relies on adjuvant surgery and radiotherapy / chemotherapy. Radiotherapy and chemotherapy are often accompanied by severe toxic side effects, and patients' quality of life has not been fundamentally improved.

[0003] Curcumin, a polyphenolic compound extracted from turmeric, has attracted widespread attention due to its multiple biological activities, including antitumor, antioxidant, and anti-inflammatory effects. However, the limited antitumor activity and bioavailability of curcumin itself in vivo restrict its clinical application. To overcome these limitations, researchers have begun to synthesize a series of curcumin analogs based on the curcumin core structure, aiming to improve bioavailability and antitumor activity. Xanthohumols are a class of secondary metabolites, mainly derived from certain molds. Recent studies have shown that they possess certain anticancer activities, but their significant toxic side effects on normal cells and tissues severely limit their practical application. Therefore, researching a benzo-α-pyran-type monocarbonyl curcumin analog and its preparation method that can improve bioavailability and antitumor activity is of great significance for the preparation of antitumor drugs. Summary of the Invention

[0004] The purpose of this invention is to provide a benzo-α-pyran-type monocarbonyl curcumin analog, its preparation method, and its application, in order to solve the problems of low bioavailability and low antitumor activity of curcumin in the prior art.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0006] This invention provides a benzo-α-pyran-type monocarbonyl curcumin analog, wherein the benzo-α-pyran-type monocarbonyl curcumin analog has the structure shown in formula (Ⅰ):

[0007]

[0008] This invention provides a method for preparing the above-described benzo-α-pyran-type monocarbonyl curcumin analog. The synthetic route for the benzo-α-pyran-type monocarbonyl curcumin analog is as follows:

[0009]

[0010] The preparation method of benzo-α-pyran-type monocarbonyl curcumin analogs includes the following steps:

[0011] (1) 3-methyl-2-butenal, calcium chloride, compound 1, triethylamine and anhydrous ethanol were mixed and subjected to a cyclization reaction to obtain compound 2;

[0012] (2) Compound 2, sodium hydroxide solution and acetone were mixed and subjected to the first Claisen-Schmidt reaction to obtain compound 3;

[0013] (3) Compound 3, o-chlorobenzaldehyde, sodium hydroxide solution and anhydrous ethanol were mixed and subjected to a second Claysen-Schmidt reaction to obtain a benzo-α-pyran type monocarbonyl curcumin analog.

[0014] Preferably, in step (1), the molar ratio of 3-methyl-2-butenal, calcium chloride, compound 1, and triethylamine is 25-30:12-16:12-16:25-30; and the molar volume ratio of compound 1 and anhydrous ethanol is 12-16 mmol:30-50 mL.

[0015] Preferably, in step (1), the cyclization reaction temperature is 60–100°C and the cyclization reaction time is 1–3 h.

[0016] Preferably, in step (2), the molar volume ratio of compound 2 to acetone is 4-7 mmol: 30-50 mL; the volume ratio of sodium hydroxide solution to acetone is 1-2 mL: 30-50 mL; and the mass concentration of sodium hydroxide solution is 15-25%.

[0017] Preferably, in step (2), the temperature of the first Claisen-Schmidt reaction is 20-30°C, and the time of the first Claisen-Schmidt reaction is 44-52 h.

[0018] Preferably, in step (3), the molar ratio of compound 3 to o-chlorobenzaldehyde is 0.6–0.9:1.5–1.8; the molar volume ratio of compound 3 to anhydrous ethanol is 0.6–0.9 mmol:40–60 mL; the molar volume ratio of compound 3 to sodium hydroxide solution is 0.6–0.9 mmol:10–20 mL; and the mass concentration of sodium hydroxide solution is 15–25%.

[0019] Preferably, in step (3), the temperature of the second Claisen-Schmidt reaction is 20-30°C, and the time of the second Claisen-Schmidt reaction is 10-14h.

[0020] This invention provides the application of the above-described benzo-α-pyran-type monocarbonyl curcumin analog in the preparation of antitumor drug formulations, wherein the tumor includes liver cancer, cervical cancer, osteosarcoma, ovarian cancer, melanoma, head and neck cancer, breast cancer, lymphoma, or leukemia.

[0021] Preferably, the antitumor drug formulation comprises injections, tablets, capsules, aerosols, suppositories, films, pellets, or ointments.

[0022] The beneficial effects of this invention are:

[0023] (1) Based on the curcumin structural skeleton, this invention synthesizes a benzo-α-pyran-type monocarbonyl curcumin analog by combining the pharmacological and medicinal groups of flavophenol compounds. This not only retains the antitumor active site of curcumin, but also improves the bioavailability of curcumin, reduces the toxic side effects on normal cells, and improves the antitumor activity, with higher stability and better antitumor activity.

[0024] (2) The benzo-α-pyran-type monocarbonyl curcumin analog synthesized in this invention has excellent anti-tumor activity against cervical cancer, liver cancer, osteosarcoma and ovarian cancer, but has virtually no toxic side effects on normal liver cells, and is expected to become a broad-spectrum anti-tumor drug. Attached Figure Description

[0025] Figure 1 The benzo-α-pyran-type monocarbonyl curcumin analog prepared in Example 1 1 H NMR spectrum;

[0026] Figure 2 The benzo-α-pyran-type monocarbonyl curcumin analog prepared in Example 1 13 C10 NMR spectrum;

[0027] Figure 3 The results of the benzo-α-pyran-type monocarbonyl curcumin analog prepared in Example 1 promoting tumor cell apoptosis;

[0028] Figure 4 The results show that the benzo-α-pyran-type monocarbonyl curcumin analog prepared in Example 1 inhibited tumor cell metastasis. Detailed Implementation

[0029] This invention provides a benzo-α-pyran-type monocarbonyl curcumin analog, wherein the benzo-α-pyran-type monocarbonyl curcumin analog has the structure shown in formula (Ⅰ):

[0030]

[0031] This invention provides a method for preparing the benzo-α-pyran-type monocarbonyl curcumin analog, and the synthetic route for the benzo-α-pyran-type monocarbonyl curcumin analog is as follows:

[0032]

[0033] The preparation method of benzo-α-pyran-type monocarbonyl curcumin analogs includes the following steps:

[0034] (1) 3-methyl-2-butenal, calcium chloride, compound 1, triethylamine and anhydrous ethanol were mixed and subjected to a cyclization reaction to obtain compound 2;

[0035] (2) Compound 2, sodium hydroxide solution and acetone were mixed and subjected to the first Claisen-Schmidt reaction to obtain compound 3;

[0036] (3) Compound 3, o-chlorobenzaldehyde, sodium hydroxide solution and anhydrous ethanol were mixed and subjected to a second Claysen-Schmidt reaction to obtain a benzo-α-pyran type monocarbonyl curcumin analog.

[0037] In this invention, in step (1), the molar ratio of 3-methyl-2-butenal, calcium chloride, compound 1, and triethylamine is 25-30:12-16:12-16:25-30, preferably 26-29:13-15:13-15:26-29, and more preferably 27-28.86:14.48:14.48:27-28.86; the molar volume ratio of compound 1 and anhydrous ethanol is 12-16 mmol:30-50 mL, preferably 13-15 mmol:35-45 mL, and more preferably 14.48 mmol:40 mL.

[0038] In this invention, in step (1), the temperature of the cyclization reaction is 60-100°C, preferably 70-90°C, and more preferably 80°C, and the time of the cyclization reaction is 1-3 hours, preferably 1.5-2.5 hours, and more preferably 2 hours.

[0039] In this invention, in step (2), the molar volume ratio of compound 2 to acetone is 4-7 mmol: 30-50 mL, preferably 5-6 mmol: 35-45 mL, more preferably 5.6-5.9 mmol: 40 mL; the volume ratio of sodium hydroxide solution to acetone is 1-2 mL: 30-50 mL, preferably 1.2-1.8 mL: 35-45 mL, more preferably 1.5 mL: 40 mL; the mass concentration of sodium hydroxide solution is 15-25%, preferably 18-22%, more preferably 20%.

[0040] In this invention, in step (2), the temperature of the first Claisen-Schmidt reaction is 20-30°C, preferably 22-28°C, and more preferably 25°C; the time of the first Claisen-Schmidt reaction is 44-52h, preferably 46-50h, and more preferably 48h.

[0041] In this invention, in step (3), the molar ratio of compound 3 to o-chlorobenzaldehyde is 0.6–0.9:1.5–1.8, preferably 0.65–0.85:1.6–1.7, and more preferably 0.7–0.82:1.64–1.66; the molar volume ratio of compound 3 to anhydrous ethanol is 0.6–0.9 mmol:40–60 mL, preferably 0.65–0.85 mmol:45–55 mL, and more preferably 0.7–0.82 mmol:50 mL; the molar volume ratio of compound 3 to sodium hydroxide solution is 0.6–0.9 mmol:10–20 mL, preferably 0.65–0.85 mmol:12–18 mL, and more preferably 0.7–0.82 mmol:15 mL; and the mass concentration of sodium hydroxide solution is 15–25%, preferably 18–22%, and more preferably 20%.

[0042] In this invention, in step (3), the temperature of the second Claisen-Schmidt reaction is 20-30°C, preferably 22-28°C, and more preferably 25°C; the time of the second Claisen-Schmidt reaction is 10-14h, preferably 11-13h, and more preferably 12h.

[0043] This invention provides the application of the above-described benzo-α-pyran-type monocarbonyl curcumin analog in the preparation of antitumor drug formulations, wherein the tumor includes liver cancer, cervical cancer, osteosarcoma, ovarian cancer, melanoma, head and neck cancer, breast cancer, lymphoma, or leukemia.

[0044] In this invention, the antitumor drug preparation comprises injections, tablets, capsules, aerosols, suppositories, films, drops, or ointments, preferably injections, tablets, capsules, aerosols, suppositories, or ointments, and more preferably injections, tablets, or capsules.

[0045] 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.

[0046] Compound 1 in the embodiments of the present invention is Compound 2 is Compound 3 is

[0047] Example 1

[0048] 28.86 mmol of 3-methyl-2-butenal, 14.48 mmol of calcium chloride, and 14.48 mmol of compound 1 were added to 40 mL of anhydrous ethanol, followed by 28.86 mmol of triethylamine. The reaction mixture was stirred at 12000 rpm and refluxed at 80 °C for 2 hours. After the reaction was complete, the reaction was quenched with water, and the solvent anhydrous ethanol was removed by vacuum distillation at 0.1 kPa. The reaction product was dissolved in distilled water (30 mL) and extracted three times with ethyl acetate (30 mL). The organic layer was dried over anhydrous magnesium sulfate, and the ethyl acetate was removed by vacuum distillation at 0.1 kPa. Finally, the product was purified by silica gel column chromatography to give compound 2 (a yellow oily liquid) in a yield of 63.9%.

[0049] 5.88 mmol of compound 2 was added to 40 mL of acetone, followed by 1.5 mL of sodium hydroxide solution (20% by mass). The mixture was stirred at 12000 rpm and reacted at 25 °C for 48 h. After the reaction was complete, the solvent acetone was removed by vacuum distillation under 0.1 kPa. The reaction product was dissolved in 30 mL of distilled water and extracted three times with ethyl acetate (30 mL of ethyl acetate each time). The organic layer was dried over anhydrous magnesium sulfate and the ethyl acetate was removed by vacuum distillation under 0.1 kPa. Finally, the product was purified by silica gel column chromatography to give compound 3 (yellow solid), with a yield of 71.5%.

[0050] 0.818 mmol of compound 3 and 1.64 mmol of o-chlorobenzaldehyde were added to 50 mL of anhydrous ethanol, followed by 15 mL of sodium hydroxide solution (20% by mass). The mixture was stirred at 12000 rpm and reacted at 25 °C for 12 h. After the reaction was complete, the product was neutralized with hydrochloric acid solution (36% concentration), then dissolved in distilled water (30 mL), and extracted three times with ethyl acetate (30 mL ethyl acetate each time). The organic layer was dried over anhydrous magnesium sulfate, and the ethyl acetate was removed by vacuum distillation at 0.1 kPa. Finally, the product was purified by silica gel column chromatography to obtain a benzo-α-pyran-type monocarbonyl curcumin analog (yellow solid) in a yield of 21.3%.

[0051] Example 2

[0052] 30 mmol of 3-methyl-2-butenal, 16 mmol of calcium chloride, and 16 mmol of compound 1 were added to 50 mL of anhydrous ethanol, followed by 30 mmol of triethylamine. The reaction mixture was stirred at 12,000 rpm and refluxed at 100 °C for 1 hour. After the reaction was complete, the reaction was quenched with water, and the solvent anhydrous ethanol was removed by vacuum distillation under 0.1 kPa. The reaction product was dissolved in distilled water (30 mL) and extracted three times with ethyl acetate (30 mL ethyl acetate each time). The organic layer was dried over anhydrous magnesium sulfate, and the ethyl acetate was removed by vacuum distillation under 0.1 kPa. Finally, the product was purified by silica gel column chromatography to give compound 2 (a yellow oily liquid) in a yield of 62.8%.

[0053] 7 mmol of compound 2 was added to 50 mL of acetone, followed by 2 mL of sodium hydroxide solution (25% by mass). The mixture was stirred at 12000 rpm and reacted at 30 °C for 44 h. After the reaction was complete, the solvent acetone was removed by vacuum distillation under 0.1 kPa. The reaction product was dissolved in 30 mL of distilled water and extracted three times with ethyl acetate (30 mL of ethyl acetate each time). The organic layer was dried over anhydrous magnesium sulfate and the ethyl acetate was removed by vacuum distillation under 0.1 kPa. Finally, the product was purified by silica gel column chromatography to give compound 3 (yellow solid), with a yield of 70.8%.

[0054] 0.9 mmol of compound 3 and 1.8 mmol of o-chlorobenzaldehyde were added to 60 mL of anhydrous ethanol, followed by 10 mL of sodium hydroxide solution (25% by mass). The mixture was stirred at 12000 rpm and reacted at 30 °C for 10 h. After the reaction was complete, the product was neutralized with hydrochloric acid solution (36% concentration), then dissolved in distilled water (30 mL), and extracted three times with ethyl acetate (30 mL ethyl acetate each time). The organic layer was dried over anhydrous magnesium sulfate, and the ethyl acetate was removed by vacuum distillation at 0.1 kPa. Finally, the product was purified by silica gel column chromatography to obtain a benzo-α-pyran-type monocarbonyl curcumin analog (yellow solid) with a yield of 22.1%.

[0055] Example 3

[0056] 25 mmol of 3-methyl-2-butenal, 12 mmol of calcium chloride, and 12 mmol of compound 1 were added to 30 mL of anhydrous ethanol, followed by 25 mmol of triethylamine. The reaction mixture was stirred at 12,000 rpm and refluxed at 60 °C for 3 hours. After the reaction was complete, the reaction was quenched with water, and the solvent anhydrous ethanol was removed by vacuum distillation at 0.1 kPa. The reaction product was dissolved in distilled water (30 mL) and extracted three times with ethyl acetate (30 mL ethyl acetate each time). The organic layer was dried over anhydrous magnesium sulfate, and the ethyl acetate was removed by vacuum distillation at 0.1 kPa. Finally, the product was purified by silica gel column chromatography to give compound 2 (a yellow oily liquid) in a yield of 64.3%.

[0057] 4 mmol of compound 2 was added to 30 mL of acetone, followed by 1 mL of sodium hydroxide solution (15% by mass). The mixture was stirred at 12000 rpm and reacted at 20 °C for 52 h. After the reaction was complete, the solvent acetone was removed by vacuum distillation under 0.1 kPa. The reaction product was dissolved in 30 mL of distilled water and extracted three times with ethyl acetate (30 mL of ethyl acetate each time). The organic layer was dried over anhydrous magnesium sulfate and the ethyl acetate was removed by vacuum distillation under 0.1 kPa. Finally, the product was purified by silica gel column chromatography to give compound 3 (yellow solid), with a yield of 70.3%.

[0058] 0.6 mmol of compound 3 and 1.5 mmol of o-chlorobenzaldehyde were added to 40 mL of anhydrous ethanol, followed by 20 mL of sodium hydroxide solution (15% by mass). The mixture was stirred at 12,000 rpm and reacted at 20 °C for 14 h. After the reaction was complete, the product was neutralized with hydrochloric acid solution (36% concentration), then dissolved in distilled water (30 mL), and extracted three times with ethyl acetate (30 mL ethyl acetate each time). The organic layer was dried over anhydrous magnesium sulfate, and the ethyl acetate was removed by vacuum distillation at 0.1 kPa. Finally, the product was purified by silica gel column chromatography to obtain a benzo-α-pyran-type monocarbonyl curcumin analog (yellow solid) in a yield of 20.7%.

[0059] The benzo-α-pyran-type monocarbonyl curcumin analog prepared in Example 1 was characterized by NMR, and the results are shown in the figure. Figures 1-2 The obtained data is as follows:

[0060] 1H-NMR (400MHz, DMSO-d6) δ (ppm): 9.80 (s, 1H), 8.04 (d, J = 16.0Hz, 1H), 8.00-7.94 (m, 1H), 7.89 (d, J = 15.8Hz, 1H), 7.57-7.48 (m, 2 H),7.46-7.34(m,3H),7.08(d,J=15.9Hz,1H),6.73(d,J=10.1Hz,1H),6.38(d,J=8.6Hz,1H),5.70(d,J=10.0Hz,1H),1.35(s,6H). 13 C-NMR(101MHz,DMSO-d6)δ(ppm):188.45,156.63,153.61,139.73,136.76,134.58,133.10,132.18,130 .58,129.52,129.19,129.10,128.66,128.27,123.70,117.08,116.34,110.63,109.77,76.74,28.06×2.

[0061] Test Example 1

[0062] The cytotoxicity of the benzo-α-pyran-type monocarbonyl curcumin analog, curcumin, and xanthohumol C prepared in Example 1 to normal and tumor cells was tested using the MTT assay.

[0063] Cell viability / toxicity assay (MTT): a) Cells in logarithmic growth phase (normal hepatocytes MIHA, cervical cancer HeLa cells, hepatocellular carcinoma HepG2 cells, osteosarcoma MG63 cells, and ovarian cancer SKOV3 cells) were seeded in 96-well plates at a density of 5000 cells / well and 160 μL / well; b) 12 hours after seeding, 40 μL of DMEM medium (purchased from Gibco) containing benzo-α-pyran-type monocarbonyl curcumin analogue, curcumin, or xanthohumol C from Example 1 was added to each well. The concentrations of benzo-α-pyran-type monocarbonyl curcumin analogue, curcumin, or xanthohumol C in the DMEM medium were 200 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM, and 1.5625 μM, respectively, to maintain a total volume of 200 μL / well. Blank wells (without MTT staining) and control wells (DMEM medium without benzo-α-pyran monocarbonyl curcumin analogue) were set up and cultured for 44 hours. c) 20 μL of MTT solution (5 mg / mL) was added to each well, and the mixture was incubated at 37°C for another 4 hours. The supernatant was then removed, and 100 μL of DMSO was added to each well. The mixture was shaken for 15 seconds to fully dissolve the formazan. d) The absorbance of each well was measured at 492 nm using a microplate reader, and the results were recorded. All experiments were repeated three times, and the average value and error were calculated. The results are shown in Table 1.

[0064] The testing procedure for the cytotoxicity of curcumin and xanthohumol C against normal and tumor cells is the same as that for benzo-α-pyran monocarbonyl curcumin analogs.

[0065] The structural formula of xanthohumol C is: Table 1. IC50 (μM) values ​​of benzo-α-pyran-type monocarbonyl curcumin analogues, curcumin, and xanthohumol C against normal hepatocytes and tumor cells in Example 1.

[0066]

[0067] Table 1 shows that curcumin exhibits comparable cytotoxicity against normal hepatocytes and tumor cells. While xanthohumol C possesses antitumor activity against tumor cells, its toxicity to normal hepatocytes is greater than that against tumor cells, thus limiting its drug development and clinical application. In contrast, the benzo-α-pyran-type monocarbonyl curcumin analog synthesized based on the curcumin skeleton and incorporating the active group of xanthohumol shows significantly lower toxicity to normal cells than curcumin, but significantly higher toxicity to tumor cells, exhibiting a clear tumor cell predisposition. The benzo-α-pyran-type monocarbonyl curcumin analog synthesized in this invention possesses stronger antitumor activity than both curcumin and xanthohumol C, while effectively reducing the toxicity of curcumin and xanthohumol C to normal cells.

[0068] As demonstrated in Test Example 1, the benzo-α-pyran-type monocarbonyl curcumin analog synthesized in this invention exhibits excellent antitumor activity against cervical cancer HeLa cells, liver cancer HepG2 cells, osteosarcoma MG63 cells, and ovarian cancer SKOV3 cells. The term "tumor" in this invention includes, but is not limited to, cervical cancer HeLa cells, liver cancer HepG2 cells, osteosarcoma MG63 cells, and ovarian cancer SKOV3 cells.

[0069] Test Example 2

[0070] Test of the effect of the benzo-α-pyran-type monocarbonyl curcumin analog obtained in Example 1 on promoting tumor cell death: The benzo-α-pyran-type monocarbonyl curcumin analog obtained in Example 1 was added to DMEM medium (purchased from Gibco) at specified concentrations, wherein the concentrations of the benzo-α-pyran-type monocarbonyl curcumin analog were 5.0 μM, 10.0 μM, and 20.0 μM.

[0071] a) The human cervical cancer cell line HeLa was seeded into a 10cm cell culture dish at a cell density of 5×10⁻⁶ cells / cm². 6 After 6 hours of cell adhesion at a density of 1 / mL, DMEM medium containing different concentrations of benzo-α-pyran-type monocarbonyl curcumin analogue was added. The control group received a corresponding volume of 0.1% DMSO in DMEM medium. Cells were cultured for another 48 hours. b) All cells (including those that floated) were collected. Adhered cells were collected using 0.25% trypsin digestion, centrifuged, and washed twice with 1×PBS. The supernatant was discarded, and the precipitate (cells) was retained. c) The collected cells were... Fluor488annexin V and PI were used for double staining, and the specific staining procedure was performed according to Alexa standards. The annexin V / Dead Cell Apoptosis Kit was used as instructed; d) the stained cell suspension was directly tested using flow cytometry, and the experimental data were processed and fitted using FlowJo software. The results are as follows: Figure 3 As shown in the scatter plot of a bivariate flow cytometer, the lower left quadrant shows live cells, the lower right quadrant shows early apoptotic cells, the upper left quadrant shows mid-term apoptotic cells, and the upper right quadrant shows late apoptotic cells.

[0072] from Figure 3 It can be seen that the benzo-α-pyran-type monocarbonyl curcumin analog synthesized in this invention can effectively induce apoptosis in the cervical cancer cell line HeLa, and the ability to induce tumor cell apoptosis is dose-dependent.

[0073] Test Example 3

[0074] The benzo-α-pyran-type monocarbonyl curcumin analog obtained in Example 1 inhibits the metastasis of tumor cells:

[0075] (1) Take 2×10 5 HeLa cells were seeded at a density of 10 cells / mL into 6-well plates. After cell adhesion, DMEM medium (purchased from Gibco) containing specified concentrations of benzo-α-pyran monocarbonyl curcumin analogues (5.0 μM, 10.0 μM, and 20.0 μM) was added. The control group was treated with the corresponding volume of 0.1% DMSO. After incubation for 48 hours, cells were digested, counted, and 1×10⁶ cells were separated. 5 (1) Mix 100 μL of serum-free culture medium and seed the cells into Transwell chambers. Add 600 μL of culture medium containing 10% serum to the bottom of the chamber. (2) Incubate for 72 h. Wipe the cells that have not penetrated the inner layer of the chamber with a cotton swab. Add 0.5 mL of 4% paraformaldehyde to each well and fix for 15 min at room temperature. (3) Wash along the side wall of the chamber with PBS and dry with a cotton swab. (4) Add 500 μL of crystal violet to each well and stain in the dark for 5 min. (5) Wash the unstained crystal violet along the side wall of the chamber with double-distilled water (dd H2O) and dry with a cotton swab. Add 500 μL of acetic acid (33% concentration) to each well to fully dissolve the crystal violet. (6) Take 100 μL of the dissolved glacial acetic acid into a 96-well plate and set up three sub-wells for each group. Detect the absorbance of each group at a wavelength of 560 nm using an ELISA reader.

[0076] Experimental results are as follows Figure 4 As shown, from Figure 4 It can be seen that, compared with the control group, the administration of benzo-α-pyran-type monocarbonyl curcumin analog can effectively inhibit the permeability of HeLa cells, and the inhibition of permeability gradually increases with the increase of concentration, indicating that the benzo-α-pyran-type monocarbonyl curcumin analog synthesized in this invention can effectively inhibit the metastatic ability of tumor cells.

[0077] As can be seen from the above embodiments, the present invention provides a benzo-α-pyran type monocarbonyl curcumin analog, its preparation method and application. Based on the curcumin skeleton, the present invention combines the pharmacological and medicinal groups of flavophenol compounds to synthesize a benzo-α-pyran type monocarbonyl curcumin analog with the structure of formula (I). It has excellent anti-tumor activity against cervical cancer, liver cancer, osteosarcoma and ovarian cancer, and has low toxicity to normal liver cells.

[0078] 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 benzo-α-pyran-type monocarbonyl curcumin analog, characterized in that, The benzo-α-pyran-type monocarbonyl curcumin analog has the structure shown in formula (Ⅰ):

2. The method for preparing the benzo-α-pyran-type monocarbonyl curcumin analog according to claim 1, characterized in that, The synthetic route for benzo-α-pyran-type monocarbonyl curcumin analogs is as follows: The preparation method of benzo-α-pyran-type monocarbonyl curcumin analogs includes the following steps: (1) 3-methyl-2-butenal, calcium chloride, compound 1, triethylamine and anhydrous ethanol were mixed and subjected to a cyclization reaction to obtain compound 2; (2) Compound 2, sodium hydroxide solution and acetone were mixed and subjected to the first Claisen-Schmidt reaction to obtain compound 3; (3) Compound 3, o-chlorobenzaldehyde, sodium hydroxide solution and anhydrous ethanol were mixed and subjected to a second Claysen-Schmidt reaction to obtain a benzo-α-pyran type monocarbonyl curcumin analog.

3. The method for preparing benzo-α-pyran-type monocarbonyl curcumin analogs according to claim 2, characterized in that, In step (1), the molar ratio of 3-methyl-2-butenal, calcium chloride, compound 1, and triethylamine is 25-30:12-16:12-16:25-30; the molar volume ratio of compound 1 and anhydrous ethanol is 12-16 mmol:30-50 mL.

4. The method for preparing benzo-α-pyran-type monocarbonyl curcumin analogs according to claim 2 or 3, characterized in that, In step (1), the cyclization reaction temperature is 60-100℃ and the cyclization reaction time is 1-3h.

5. The method for preparing benzo-α-pyran-type monocarbonyl curcumin analogs according to claim 4, characterized in that, In step (2), the molar volume ratio of compound 2 to acetone is 4-7 mmol: 30-50 mL; the volume ratio of sodium hydroxide solution to acetone is 1-2 mL: 30-50 mL; and the mass concentration of sodium hydroxide solution is 15-25%.

6. The method for preparing benzo-α-pyran-type monocarbonyl curcumin analogs according to claim 3 or 5, characterized in that, In step (2), the temperature of the first Claisen-Schmidt reaction is 20-30°C, and the time of the first Claisen-Schmidt reaction is 44-52 h.

7. The method for preparing benzo-α-pyran-type monocarbonyl curcumin analogs according to claim 6, characterized in that, In step (3), the molar ratio of compound 3 to o-chlorobenzaldehyde is 0.6–0.9:1.5–1.8; the molar volume ratio of compound 3 to anhydrous ethanol is 0.6–0.9 mmol:40–60 mL; the molar volume ratio of compound 3 to sodium hydroxide solution is 0.6–0.9 mmol:10–20 mL; and the mass concentration of sodium hydroxide solution is 15–25%.

8. The method for preparing benzo-α-pyran-type monocarbonyl curcumin analogs according to claim 5 or 7, characterized in that, In step (3), the temperature of the second Claisen-Schmidt reaction is 20-30°C, and the time of the second Claisen-Schmidt reaction is 10-14 hours.

9. The use of the benzo-α-pyran-type monocarbonyl curcumin analogue according to claim 1 in the preparation of antitumor drug formulations, characterized in that, The tumor is liver cancer, cervical cancer, osteosarcoma, or ovarian cancer.

10. The application according to claim 9, characterized in that, The antitumor drug preparation is an injection, tablet, capsule, aerosol, suppository, film, drop pill, or ointment.

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