Polyphenol derivative as well as preparation method and application thereof

Abstract

The invention discloses a polyphenol derivative as well as a preparation method and application thereof. Pharmacological studies prove that the polyphenol derivative L96-2 provided by the invention can effectively inhibit proliferation of human breast cancer cell lines MCF-7 and MDA-MB-231 and a human nasopharyngeal carcinoma cell line C666-1, and the inhibitory activity of the polyphenol derivative L96-2 is obviously higher than that of parent compounds icaritin and isatin of the polyphenol derivative L96-2. Meanwhile, the L96-2 can be used for effectively inducing apoptosis of the MCF-7 cells. The polyphenol derivative L96-2 provided by the invention has a very good inhibition effect on breast cancer and nasopharynx cancer cells and has a very good development value, and the technology also provides a new thought and approach for synthesis and antitumor research of polyphenol derivatives. The preparation method provided by the invention has the advantages of simplicity, cheap and easily available raw materials, no special requirements on production equipment, easiness in operation, low cost and easiness in realization of industrial production.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a polyphenol derivative, its preparation method, and its application. Background Technology

[0002] Malignant tumors are common and prevalent diseases that seriously threaten human health and life. Breast cancer, as the malignant tumor with the highest incidence and mortality rate among women worldwide, has always received widespread attention. Breast cancer has a high incidence, recurrence rate, and mortality rate, making early diagnosis and treatment crucial. Currently, clinical treatment for breast cancer mainly involves surgery and chemotherapy, but these often come with a series of complications, such as postoperative depression, severely impacting patients' survival rate and quality of life. Nasopharyngeal carcinoma is a malignant tumor originating in the top and lateral walls of the nasopharynx, exhibiting a distinct geographical distribution, primarily originating in southern my country. Due to the close proximity of the nasopharynx to brain stem cell regions, major blood vessels, and nerves, coupled with the often subtle early symptoms of nasopharyngeal carcinoma, patients are usually diagnosed at an advanced stage, making surgical treatment difficult. Radiotherapy and chemotherapy are the main treatment methods, but these are prone to complications affecting the ear, nose, and brain.

[0003] Polyphenols are a collective term for plant secondary metabolites containing multiple phenolic hydroxyl groups in their molecular structure. They are one of the most important families of natural products, widely found in fruits, vegetables, tea, legumes, grains, and other plants. Phenylacetin and flavonoids are examples of polyphenols. Polyphenolic compounds possess various biological activities, including anti-inflammatory and antioxidant effects. Icaritin is a natural polyphenol extracted from the traditional Chinese medicine Epimedium. Icaritin exhibits multiple biological activities, including anti-inflammatory, antioxidant, and antitumor effects; however, its poor water solubility and low oral bioavailability limit its clinical efficacy. Therefore, researchers are working to improve these issues through strategies such as structural modification. Indole-2,3-dione is a natural indole analog with various biological activities, including antibacterial and antitumor effects. Although indole and its derivatives have shown good potential in antitumor research, their selectivity for normal cells is low. The inventor has been dedicated to the research of small molecule drugs based on natural products, and has carried out a number of drug modification studies on polyphenols and indigo compounds, resulting in a novel polyphenol derivative L96-2, which has no relevant literature reports to date. Summary of the Invention

[0004] The purpose of this invention is to provide a polyphenol derivative and its preparation method to address the shortcomings of existing technologies. This compound has a good inhibitory effect on breast cancer and nasopharyngeal carcinoma cells, which is significantly stronger than its parent compounds icariin and indigo, and has great development value. This technology also provides new ideas and approaches for the synthesis and anti-tumor research of polyphenol derivatives, and the preparation process is simple and the raw materials are easy to obtain.

[0005] The technical solution of the present invention is as follows:

[0006] A polyphenol derivative, characterized by the following structural formula:

[0007]

[0008] The preparation method of the polyphenol derivatives as described above includes the following steps:

[0009] (1) Take indigo, add N,N-dimethylformamide, stir to dissolve, and set aside;

[0010] (2) Add cesium carbonate and 1,4-dibromobutane to the solution in step (1) in sequence, stir the reaction, quench the reaction with water, extract with ethyl acetate, and combine the organic phases;

[0011] (3) Take the organic phase from step (2), wash it with water, then wash it with saturated saline solution, separate the organic phase, dry it with anhydrous Na2SO4, filter it, and keep the filtrate for later use.

[0012] (4) Take the filtrate from step (3), concentrate it, load it onto a silica gel column, elute, collect the eluent, concentrate it, and obtain a red solid.

[0013] (5) Add acetone solution of epimedium to the red solid obtained in step (4), heat to reflux, cool to room temperature, quench the reaction with water, extract with ethyl acetate, combine the organic phases, and set aside for use;

[0014] (6) Take the organic phase from step (5), wash it with saturated brine, separate the organic phase, dry it with anhydrous Na2SO4, filter it, and keep the filtrate for later use.

[0015] (7) Take the filtrate from step (6), concentrate it, load it onto a silica gel column, elute, collect the eluent, concentrate it, and obtain a yellow liquid, which is the polyphenol derivative.

[0016] Furthermore, in the preparation method of the polyphenol derivatives described above, the amount of indigo used in step (1) is 1.9g, and the amount of N,N-dimethylformamide added is 45mL.

[0017] Furthermore, in the preparation method of the polyphenol derivatives described above, the amount of cesium carbonate added in step (2) is 6.5g, the amount of 1,4-dibromobutane added is 4.9mL, the stirring reaction temperature is 25℃, the reaction time is 24 hours, the amount of water added for quenching the reaction is 20mL, and the extraction with ethyl acetate is performed 3 times with 20mL each time.

[0018] Furthermore, in the preparation method of the polyphenol derivatives described above, the washing with water in step (3) involves washing with water 7 times, 15 mL each time, and using 20 mL of saturated saline solution.

[0019] Furthermore, in the preparation method of the polyphenol derivatives described above, the eluent in step (4) is a solution with a volume ratio of petroleum ether: ethyl acetate = 5:1.

[0020] Furthermore, in the preparation method of the polyphenol derivatives described above, the acetone solution of icariin in step (5) is obtained by dissolving 626 mg of icariin in 17 mL of acetone, the heating and reflux is carried out at 65 °C for 40 hours, the amount of water added to quench the reaction is 20 mL, and the extraction with ethyl acetate is carried out by extracting with ethyl acetate 3 times, 20 mL each time.

[0021] Furthermore, in the preparation method of the polyphenol derivatives described above, the amount of saturated saline solution used in step (6) is 20 mL.

[0022] Furthermore, in the preparation method of the polyphenol derivatives described above, the eluent in step (7) is ethyl acetate.

[0023] The application of the polyphenol derivatives as described in claim 1 is in the preparation of drugs for treating breast cancer and nasopharyngeal carcinoma.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] 1. This invention uses icariin as the parent compound and achieves the first chemical splicing of icariin-indoleone through chemical methods, resulting in a novel polyphenol derivative L96-2.

[0026] 2. Pharmacological studies have confirmed that the polyphenol derivative L96-2 obtained in this invention can effectively inhibit the proliferation of human breast cancer cell lines MCF-7 and MDA-MB-231, as well as human nasopharyngeal carcinoma cell line C666-1. Its inhibitory activity is significantly stronger than that of its parent compounds, icariin and indigo. Simultaneously, L96-2 can effectively induce apoptosis in MCF-7 cells. This indicates that L96-2 provided by this invention has a good inhibitory effect on breast cancer and nasopharyngeal carcinoma cells, and has significant development value. This technology also provides new ideas and approaches for the synthesis and anti-tumor research of polyphenol derivatives.

[0027] 3. The preparation method of this invention is simple, the raw materials are inexpensive and readily available, there are no special requirements for production equipment, it is easy to operate, low in cost, and easy to realize industrial production. Attached Figure Description

[0028] Figure 1 For L96-2 1 HNMR spectrum (400M, solvent: deuterated chloroform).

[0029] Figure 2 For L96-2 13 CNMR spectrum (100M, solvent: deuterated chloroform).

[0030] Figure 3 This is the mass spectrum (ESI) of L96-2.

[0031] Figure 4 Flow cytometry plots of apoptosis in MCF-7 cells treated with different concentrations of L96-2.

[0032] Figure 5 Histograms of apoptosis in MCF-7 cells treated with different concentrations of L96-2 were plotted, n=3. Compared with the control group, ****P<0.0001, ***P<0.001. Detailed Implementation

[0033] The present invention will be further described below with reference to specific embodiments, but this does not limit the scope of protection and application of the present invention:

[0034] I. Preparation of a polyphenol derivative

[0035] Example 1

[0036] The preparation of a polyphenol derivative includes the following steps:

[0037]

[0038] (1) Take indigo (1.9g), add N,N-dimethylformamide (45mL), stir to dissolve, and set aside;

[0039] (2) Add cesium carbonate (6.5 g) and 1,4-dibromobutane (4.9 mL) to the solution in step (1) in sequence. After stirring at 25 °C for 24 hours, add water (20 mL) to quench the reaction. Extract with ethyl acetate 3 times, 20 mL each time, and combine the organic phases.

[0040] (3) Take the organic phase from step (2), wash it with water 7 times, 15 mL each time, and then wash it with saturated saline (20 mL). Separate the organic phase, dry it with anhydrous Na2SO4, filter it, and keep the filtrate for later use.

[0041] (4) Take the filtrate from step (3), concentrate it, and then elute it onto a silica gel column with a petroleum ether:ethyl acetate ratio of 5:1 (v / v). Collect the eluent, concentrate it, and obtain a red solid, R. f =0.42 (petroleum ether / ethyl acetate, 3:1);

[0042] (5) Add 17 mL of acetone solution of 626 mg of epimedium to the red solid obtained in step (4), heat under reflux at 65°C for 40 hours, cool to room temperature, add water (20 mL) to quench the reaction, extract with ethyl acetate 3 times, 20 mL each time, combine the organic phases and set aside for use;

[0043] (6) Take the organic phase from step (5), wash it with saturated saline (20 mL), separate the organic phase, dry it with anhydrous Na2SO4, filter it, and keep the filtrate for later use.

[0044] (7) Take the filtrate from step (6), concentrate it, load it onto a silica gel column, elute with ethyl acetate, collect the eluent, concentrate it, and obtain a yellow liquid L96-2 (1.1 g, yield 70%). f =0.5 (petroleum ether / ethyl acetate, 1:3).

[0045] The structural confirmation results of the prepared compound L96-2 are shown in the figure. Figure 1 — Figure 3 .

[0046] 1 H NMR (400MHz, CHLOROFORM-D) δ12.70 (s, 1H), 8.14–7.95 (m, 2H), 7.35 (ddd, J=7.4, 3.4, 1.2Hz, 2H), 7.29 (tdd, J=7.8, 2.8, 1.3Hz, 2H),7.06–7.02(m,2H),7.00–6.95(m,2H),6.88(dd,J=31.4,7.8Hz,2H),6.34(s,1H),5.18–5.10(m,1H),4.41(d,J=32.5Hz,2H), 4.08(d,J=4.9Hz,2H),3.98(t,J=5.9Hz,2H),3.86(s,3H),3.82–3.66(m,4H),3.44(d,J=6.9Hz,2H),3.21(t,J=16.8Hz,2H),2.99 (dd,J=20.4,17.0Hz,2H),2.18–2.09(m,6H),1.95–1.89(m,4H),1.88–1.80(m,4H),1.73(d,J=1.4Hz,3H),1.64(d,J=1.4Hz,3H). 13CNMR(100MHz,CHLOROFORM-D)δ207.51,207.20,179.26,176.49,176.32,161.95,161.65,160.30,156.4 0,153.55,143.02,142.94,137.54,131.85,130.37,130.14,130.10,129.91,124.07,123.98,123.27,12 3.16,123.03,122.46,114.09,109.20,108.87,107.79,105.57,95.55,74.17,74.06,72.05,68.10,55. 54,49.20,49.04,39.87,39.67,31.45,31.35,27.48,26.50,25.86,23.94,21.80,18.13.MS(ESI)[M+Na] + :909.8.

[0047] II. Pharmacological and Efficacy Tests

[0048] Example 2

[0049] Effect of L96-2 on the proliferation of human breast cancer cell line MCF-7

[0050] Objective: To investigate the effect of L96-2 on the proliferation of human breast cancer cell line MCF-7.

[0051] 1. Experimental Materials

[0052] 1.1 Cell line: Human breast cancer cell line MCF-7

[0053] 1.2 Reagents and Instruments: Fetal bovine serum (FBS, Gibco), DMEM medium (Gibco), DMSO (Solarbio), PBS buffer (Solarbio), CCK8 kit (Zeta Life), trypsin (Gibco), BioTek Synergy microplate reader (BioTek), low-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instruments Co., Ltd.), constant temperature water bath (Shanghai Yiheng Scientific Instruments Co., Ltd.), clean bench (Shanghai Yiheng Scientific Instruments Co., Ltd.), cell culture incubator (Thermo), inverted microscope (Leica).

[0054] 2. Experimental Methods

[0055] 2.1 Culture MCF-7 cells. When they reach the logarithmic growth phase, digest them with 0.25% trypsin, centrifuge at 1000 rpm for 5 min, resuspend the cell pellet in DMEM medium containing 10% FBS, and seed them into 96-well culture plates at a density of 8000 cells / well. Then, incubate overnight at 37°C, 5% CO2 and saturated humidity.

[0056] 2.2 Each group has 3 replicates. Add the drug to each well according to the following conditions, and then continue to incubate for 24 hours.

[0057] Control group: the group that did not receive any medication.

[0058] Positive control group 1: Icariin was added, with concentration gradients of 64μM, 32μM, 16μM, 8μM, 4μM, 2μM, and 1μM.

[0059] Positive control group 2: Indigo was added, with concentration gradients of 64 μM, 32 μM, 16 μM, 8 μM, 4 μM, 2 μM, and 1 μM.

[0060] L96-2 group: L96-2 was added, and the concentration gradient was set to 64μM, 32μM, 16μM, 8μM, 4μM, 2μM, and 1μM.

[0061] Twenty-four hours after the above cell intervention, the effect of the drug on tumor cell proliferation was detected using a CCK8 assay kit. The specific procedure was as follows: 10 μL of CCK8 reagent was added to each well, and the cells were incubated in the dark for 1 hour. The absorbance (OD) value was then measured at 450 nm using a microplate reader. A dose-response curve was fitted using GraphPad Prism software, and the drug concentration at which 50% cell viability was inhibited was calculated, i.e., the IC50 concentration. 50 value.

[0062] 3. Experimental Results

[0063] Table 1. Effects of L96-2 on the proliferation of human breast cancer cell line MCF-7 ( n=3)

[0064]

[0065] As shown in Table 1, L96-2 significantly inhibited the proliferation of human breast cancer cell line MCF-7, and its inhibitory effect was significantly better than that of the positive control drugs icariin and indigo.

[0066] Example 3

[0067] Effect of L96-2 on the proliferation of human breast cancer cell line MDA-MB-231

[0068] Objective: To investigate the effect of L96-2 on the proliferation of human breast cancer cell line MDA-MB-231.

[0069] 1. Experimental Materials

[0070] 1.1 Cell line: Human breast cancer cell line MDA-MB-231

[0071] 1.2 Reagents and Instruments: Fetal bovine serum (FBS, Gibco), DMEM medium (Gibco), DMSO (Solarbio), PBS buffer (Solarbio), CCK8 kit (Zeta Life), trypsin (Gibco), BioTek Synergy microplate reader (BioTek), low-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instruments Co., Ltd.), constant temperature water bath (Shanghai Yiheng Scientific Instruments Co., Ltd.), clean bench (Shanghai Yiheng Scientific Instruments Co., Ltd.), cell culture incubator (Thermo), inverted microscope (Leica).

[0072] 2. Experimental Methods

[0073] 2.1 Culture MDA-MB-231 cells until they reach the logarithmic growth phase, then... . Digest with 25% trypsin, centrifuge at 1000 rpm for 5 min, resuspend the cell pellet in DMEM medium containing 10% FBS, seed at a density of 8000 cells / well in 96-well culture plates, and then incubate overnight at 37°C, 5% CO2 and saturated humidity.

[0074] 2.2 Each group has 3 replicates. Add the drug to each well according to the following conditions, and then continue to incubate for 24 hours.

[0075] Control group: the group that did not receive any medication.

[0076] Positive control group 1: Icariin was added, with concentration gradients of 64μM, 32μM, 16μM, 8μM, 4μM, 2μM, and 1μM.

[0077] Positive control group 2: Indigo was added, with concentration gradients of 64 μM, 32 μM, 16 μM, 8 μM, 4 μM, 2 μM, and 1 μM.

[0078] L96-2 group: L96-2 was added, and the concentration gradient was set to 64μM, 32μM, 16μM, 8μM, 4μM, 2μM, and 1μM.

[0079] Twenty-four hours after the above cell intervention, the effect of the drug on tumor cell proliferation was detected using a CCK8 assay kit. The specific procedure was as follows: 10 μL of CCK8 reagent was added to each well, and the cells were incubated in the dark for 1 hour. The absorbance (OD) value was then measured at 450 nm using a microplate reader. A dose-response curve was fitted using GraphPad Prism software, and the drug concentration at which 50% cell viability was inhibited was calculated, i.e., the IC50 concentration. 50 value.

[0080] 3. Experimental Results

[0081] Table 2. Effects of L96-2 on the proliferation of human breast cancer cell line MDA-MB-231 ( n=3)

[0082]

[0083] As shown in Table 2, L96-2 significantly inhibited the proliferation of human breast cancer cell line MDA-MB-231, and its inhibitory effect was significantly better than that of the positive control drugs icariin and indigo.

[0084] Example 4

[0085] Effect of L96-2 on the proliferation of human nasopharyngeal carcinoma cell line C666-1

[0086] Objective: To investigate the effect of L96-2 on the proliferation of human nasopharyngeal carcinoma cell line C666-1.

[0087] 1. Experimental Materials

[0088] 1.1 Cell line: Human nasopharyngeal carcinoma cell line C666-1

[0089] 1.2 Reagents and Instruments: Fetal bovine serum (FBS, Gibco), RPMI Medium 1640 medium (Gibco), DMSO (Solarbio), PBS buffer (Solarbio), CCK8 kit (Zeta Life), trypsin (Gibco), BioTek Synergy microplate reader (BioTek), low-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instruments Co., Ltd.), constant temperature water bath (Shanghai Yiheng Scientific Instruments Co., Ltd.), clean bench (Shanghai Yiheng Scientific Instruments Co., Ltd.), cell culture incubator (Thermo), inverted microscope (Leica).

[0090] 2. Experimental Methods

[0091] 2.1 C666-1 cells were cultured and digested with 0.25% trypsin when they reached the logarithmic growth phase. The cells were centrifuged at 1000 rpm for 5 min, and the cell pellet was resuspended in RPMI Medium 1640 medium containing 10% FBS. The cells were then seeded at a density of 8000 cells / well in 96-well culture plates and incubated overnight at 37°C, 5% CO2 and saturated humidity.

[0092] 2.2 Each group has 3 replicates. Add the drug to each well according to the following conditions, and then continue to incubate for 24 hours.

[0093] Control group: the group that did not receive any medication.

[0094] Positive control group 1: Icariin was added, with concentration gradients of 64μM, 32μM, 16μM, 8μM, 4μM, 2μM, and 1μM.

[0095] Positive control group 2: Indigo was added, with concentration gradients of 64 μM, 32 μM, 16 μM, 8 μM, 4 μM, 2 μM, and 1 μM.

[0096] L96-2 group: L96-2 was added, and the concentration gradient was set to 64μM, 32μM, 16μM, 8μM, 4μM, 2μM, and 1μM.

[0097] Twenty-four hours after the above cell intervention, the effect of the drug on tumor cell proliferation was detected using a CCK8 assay kit. The specific procedure was as follows: 10 μL of CCK8 reagent was added to each well, and the cells were incubated in the dark for 1 hour. The absorbance (OD) value was then measured at 450 nm using a microplate reader. A dose-response curve was fitted using GraphPad Prism software, and the drug concentration at which 50% cell viability was inhibited was calculated, i.e., the IC50 concentration. 50 value.

[0098] 3. Experimental Results

[0099] Table 3. Effects of L96-2 on the proliferation of human nasopharyngeal carcinoma cell line C666-1 ( n=3)

[0100]

[0101] As shown in Table 3, L96-2 significantly inhibited the proliferation of human nasopharyngeal carcinoma cell line C666-1, and its inhibitory effect was significantly better than that of the positive control drugs icariin and indigo.

[0102] In summary, L96-2 prepared in this invention can inhibit the proliferation of various tumor cells, and its antitumor activity is significantly better than that of the positive controls icariin and indigo. It is an antitumor compound with development potential and can be directly used for the treatment of related diseases and the preparation of related drugs.

[0103] Example 5

[0104] Effect of L96-2 on apoptosis rate of human breast cancer cell line MCF-7

[0105] Objective: To investigate the effect of L96-2 on the apoptosis rate of human breast cancer cell line MCF-7.

[0106] 1. Experimental Materials

[0107] 1.1 Cell line: Human breast cancer cell line MCF-7

[0108] 1.2 Reagents and Instruments: Fetal bovine serum (FBS, Gibco), DMEM medium (Gibco), DMSO (Solarbio), PBS buffer (Solarbio), Annexin V-FITC apoptosis detection kit (Beyotime Biotech), trypsin (Gibco), BioTek Synergy microplate reader (BioTek), low-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instruments Co., Ltd.), constant temperature water bath (Shanghai Yiheng Scientific Instruments Co., Ltd.), clean bench (Shanghai Yiheng Scientific Instruments Co., Ltd.), cell culture incubator (Thermo), inverted microscope (Leica), flow cytometer (Beijing Cenglang Biotechnology Co., Ltd.).

[0109] 2. Experimental Methods

[0110] 2.1 Culture MCF-7 cells. When they reach the logarithmic growth phase, digest them with 0.25% trypsin, centrifuge at 1000 rpm for 5 min, resuspend the cell pellet in DMEM medium containing 10% FBS, and seed them into 6-well culture plates at a density of 500,000 cells / well. Then, incubate overnight at 37°C, 5% CO2 and saturated humidity.

[0111] 2.2 Each group has 3 replicates. Add the drug to each well according to the following conditions, and then continue to incubate for 24 hours.

[0112] Control group: the group that did not receive any medication.

[0113] Group 1: Add L96-2 at a concentration of 10 μM.

[0114] Group 2: Add L96-2 at a concentration of 20 μM.

[0115] Group 3: Add L96-2 at a concentration of 30 μM.

[0116] Twenty-four hours after the above cell intervention, the effect of L96-2 on the apoptosis rate of human breast cancer cell line MCF-7 was detected using the Beyotime Annexin V-FITC apoptosis detection kit. The specific procedure was as follows: Cells were digested with EDTA-free protease. Once the cells became rounded and detached from the bottom, serum-containing medium was immediately added to terminate the digestion, followed by centrifugation at 1000 rpm for 5 min. Cells were washed with pre-cooled PBS and centrifuged again at 1000 rpm for 5 min. The supernatant was discarded, and this procedure was repeated twice. Cells were gently resuspended in 195 μL of Annexin V-FITC binding buffer, followed by the addition of 5 μL of Annexin V-FITC staining solution and 10 μL of LPI staining solution, and gently mixed. The cells were incubated at room temperature in the dark for 30 min, then placed on ice. Flow cytometry was immediately used for detection, and the results were analyzed using Flowjo software (version 9.0.0, USA).

[0117] 3. Experimental Results

[0118] See Figure 4 , Figure 5 .

[0119] like Figure 4 , Figure 5 As shown, compared with the control group, L96-2 treatment significantly increased both early and late apoptotic cells in MCF-7 cells, and the apoptosis rate increased with increasing dosage.

[0120] Experimental results showed that L96-2 treatment significantly induced apoptosis in MCF-7 cells. Compared with the control group, the proportions of both early and late apoptotic cells in L96-2-treated MCF-7 cells were significantly increased, and this pro-apoptotic effect exhibited a clear dose-dependent characteristic. These results demonstrate that L96-2 can effectively induce apoptosis in MCF-7 cells, and its induction efficacy increases with increasing treatment concentration, indicating that L96-2 has potential application value in the field of anti-tumor research.

Claims

1. A polyphenol derivative, characterized in that, The structural formula is as follows:

2. The method for preparing polyphenol derivatives as described in claim 1, characterized in that, Includes the following steps: (1) Take indigo, add N,N-dimethylformamide, stir to dissolve, and set aside; (2) Add cesium carbonate and 1,4-dibromobutane to the solution in step (1) in sequence, stir the reaction, quench the reaction with water, extract with ethyl acetate, and combine the organic phases; (3) Take the organic phase from step (2), wash it with water, then wash it with saturated saline solution, separate the organic phase, dry it with anhydrous Na2SO4, filter it, and keep the filtrate for later use. (4) Take the filtrate from step (3), concentrate it, load it onto a silica gel column, elute, collect the eluent, concentrate it, and obtain a red solid. (5) Add acetone solution of epimedium to the red solid obtained in step (4), heat to reflux, cool to room temperature, quench the reaction with water, extract with ethyl acetate, combine the organic phases, and set aside for use; (6) Take the organic phase from step (5), wash it with saturated brine, separate the organic phase, dry it with anhydrous Na2SO4, filter it, and keep the filtrate for later use. (7) Take the filtrate from step (6), concentrate it, load it onto a silica gel column, elute, collect the eluent, concentrate it, and obtain a yellow liquid, which is the polyphenol derivative.

3. The method for preparing polyphenol derivatives according to claim 2, characterized in that, In step (1), the amount of indigo used is 1.9g, and the amount of N,N-dimethylformamide added is 45mL.

4. The method for preparing polyphenol derivatives according to claim 2, characterized in that, In step (2), the amount of cesium carbonate added is 6.5g, the amount of 1,4-dibromobutane added is 4.9mL, the stirring reaction temperature is 25℃, the reaction time is 24 hours, the amount of water added for quenching the reaction is 20mL, and the extraction with ethyl acetate is performed 3 times, with 20mL each time.

5. The method for preparing polyphenol derivatives according to claim 2, characterized in that, The washing with water in step (3) involves washing with water 7 times, 15 mL each time, and using 20 mL of saturated saline solution.

6. The method for preparing polyphenol derivatives according to claim 2, characterized in that, The eluent in step (4) is a solution with a volume ratio of petroleum ether: ethyl acetate = 5:

1.

7. The method for preparing polyphenol derivatives according to claim 2, characterized in that, The acetone solution of icariin in step (5) is obtained by dissolving 626 mg of icariin in 17 mL of acetone, heating to reflux at 65 °C for 40 hours, adding 20 mL of water to quench the reaction, and extracting with ethyl acetate three times, 20 mL each time.

8. The method for preparing polyphenol derivatives according to claim 2, characterized in that, The amount of saturated saline solution used in step (6) is 20 mL.

9. The method for preparing polyphenol derivatives according to claim 2, characterized in that, The eluent in step (7) is ethyl acetate.

10. The application of the polyphenol derivative as described in claim 1, characterized in that, The application is in the preparation of drugs for treating breast cancer and nasopharyngeal carcinoma.

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