A preparation method and application of polyphenol metal coordination compound

Abstract

The invention discloses a preparation method and application of a polyphenol metal coordination compound, belonging to the field of biomedicine, and the preparation method comprises the following steps: preparation of ellagic acid-copper organic framework material, preparation of ellagic acid-copper-adriamycin complex, and preparation of ellagic acid-copper-adriamycin-chondroitin sulfate complex. The invention finally prepares ellagic acid-copper-adriamycin-chondroitin sulfate complex by coordinating ellagic acid with copper ion, adriamycin and chondroitin sulfate, wherein the preparation of ellagic acid-copper-adriamycin complex and ellagic acid-copper-adriamycin-chondroitin sulfate complex have good killing effect on tumor cells, and reverse the drug resistance mechanism of tumor cells, solving the problem that adriamycin causes drug resistance in some tumors.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a preparation method and application of polyphenol metal coordination compounds. Background Art

[0002] In 2020, breast cancer surpassed lung cancer with 2.26 million new cases, becoming the malignant tumor with the highest incidence, accounting for 11.7% of all new cancer patients. Surgical resection and radiotherapy and chemotherapy are currently the main means of treating breast cancer, but breast cancer patients often develop drug resistance after long-term medication, which limits the effectiveness of current breast cancer treatment. The heterogeneity of tumor cells is one of the important reasons for drug resistance, so treatment strategies based on a single mechanism often induce drug resistance. Doxorubicin (DOX), as one of the representative drugs of anthracyclines, has become one of the most commonly used chemotherapy drugs in the treatment of breast cancer due to its potent anticancer effects. DOXIL, a passive targeted drug delivery system for doxorubicin delivery ® (Doxorubicin hydrochloride liposome injection) was approved by the FDA in 1995 for the treatment of malignant tumors. However, there are literature reports that breast cancer cells treated with DOX can develop drug resistance by activating cell stemness-related pathways including Notch and Hedgehog, as well as cell survival pathways including AKT and MAPK. Therefore, the development of new drug combination strategies and new drug formulations targeting the drug resistance mechanism of tumor cells has important potential clinical application value for overcoming chemotherapy resistance.

[0003] Metal-organic frameworks (MOFs) are a type of organic-inorganic hybrid materials generated by the coordination of metal clusters or metal ions and organic ligands. They have large pore volume, high specific surface area and easily adjustable skeleton structure, which can achieve high drug loading. However, there are no reports on the formation of complexes between metal-organic frameworks and ellagic acid to induce tumor cell death. Summary of the invention

[0004] In the first aspect of the present invention, in order to solve the problem that doxorubicin may cause tumor cells to develop drug resistance, a method for preparing a polyphenol metal coordination compound is provided, wherein the preparation steps include:

[0005] S1: adding ellagic acid to a sodium hydroxide solution, stirring, adding a copper ion solution, stirring, centrifuging, and lyophilizing the supernatant to obtain an ellagic acid-copper organic framework material;

[0006] S2: mixing ellagic acid-copper organic framework material and doxorubicin, adding water to dissolve, stirring and centrifuging, collecting precipitates, washing with water and freeze-drying to obtain ellagic acid-copper-doxorubicin complex;

[0007] S3: dispersing the ellagic acid-copper-doxorubicin complex in the chondroitin sulfate aqueous solution, stirring and centrifuging, collecting the precipitate, washing with water and freeze-drying to obtain the ellagic acid-copper-doxorubicin-chondroitin sulfate complex.

[0008] The present invention coordinates ellagic acid with copper ions to innovatively synthesize ellagic acid-copper organic framework materials, which are structurally stable under neutral physiological conditions, but can be degraded under a low pH value environment in tumor cells. Therefore, ellagic acid-copper organic framework materials can be used as potential anti-tumor drug carriers to achieve high drug loading and pH-responsive drug release at tumor sites. And through a one-pot method, ellagic acid-copper organic framework materials and doxorubicin are synthesized into ellagic acid-copper-doxorubicin complex nanoparticles. The present invention coordinates and disperses ellagic acid-copper-doxorubicin complex nanoparticles to obtain ellagic acid-copper-doxorubicin-chondroitin sulfate complexes.

[0009] Preferably, the concentration of the sodium hydroxide solution in step S1 is 0.1-0.5 M.

[0010] Preferably, the copper ion solution in step S1 is one or more of a copper sulfate solution and a copper chloride solution.

[0011] Preferably, the copper ion concentration in the copper ion solution in step S1 is 10-50 mM.

[0012] Preferably, in step S1, the molar ratio of ellagic acid to copper ions is 1:(0.5-2).

[0013] Copper, as a common MOF assembly material, can promote copper cell death in tumor cells by interfering with the tricarboxylic acid cycle process in mitochondrial respiration, providing a new potential therapeutic strategy for tumor treatment. In addition, copper ions can also reverse tumor resistance by disrupting mitochondrial energy metabolism and reducing ATP-dependent p-gp drug efflux.

[0014] Preferably, in step S2, the mass ratio of ellagic acid-copper organic framework material to doxorubicin is 1:(0.5-2).

[0015] MOF materials containing copper ions can be used in combination with chemotherapy drugs including doxorubicin during tumor treatment, which may improve the tumor resistance caused by a single chemotherapy drug. The copper death killing mechanism produced during the treatment process and the characteristics of activating immunotherapy will help improve the problem of tumor chemotherapy resistance.

[0016] Preferably, the concentration of the chondroitin sulfate aqueous solution in step S3 is 1-5 mg / mL.

[0017] Preferably, the mass ratio of chondroitin sulfate to ellagic acid-copper-doxorubicin complex in step S3 is 1:(5-10). Chondroitin sulfate (CS) is a proteoglycan widely distributed on the cell surface and can be covalently linked to proteins. Due to its high specific expression on the tumor surface, it can be used as a packaging material for active targeted delivery of tumors.

[0018] The second aspect of the present invention provides an application of a polyphenol metal coordination compound. Further, the application is to use the aforementioned polyphenol metal coordination compound to prepare a drug for treating cancer. Specifically, the application is to use the polyphenol metal coordination compound to kill tumor cells and reverse the doxorubicin resistance of tumor cells.

[0019] Preferably, the cancer is breast cancer.

[0020] Beneficial effects of the present invention: The present invention provides a method for preparing a polyphenol metal coordination compound and its application, comprising the following steps: preparing ellagic acid-copper organic framework material, preparing ellagic acid-copper-doxorubicin complex, and preparing ellagic acid-copper-doxorubicin-chondroitin sulfate complex. The present invention prepares ellagic acid-copper-doxorubicin-chondroitin sulfate complex by combining ellagic acid with copper ions, doxorubicin, and chondroitin sulfate, wherein the preparation of ellagic acid-copper-doxorubicin complex and ellagic acid-copper-doxorubicin-chondroitin sulfate complex have good killing effect on tumor cells, and reverse the drug resistance mechanism of tumor cells, solving the problem that doxorubicin causes drug resistance in some tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a flow chart for preparing the ellagic acid-copper-doxorubicin-chondroitin sulfate complex in Example 1 of the present invention;

[0022] Figure 2 The particle size measurement results and photos of the ellagic acid-copper organic framework material complex in Example 2 of the present invention;

[0023] Figure 3 The particle size measurement results and photos of the ellagic acid-copper-doxorubicin complex in Example 2 of the present invention;

[0024] Figure 4 The particle size measurement results and photos of the ellagic acid-copper-doxorubicin-chondroitin sulfate complex in Example 2 of the present invention;

[0025] Figure 5 The zeta potential measurement results of the ellagic acid-copper-doxorubicin-chondroitin sulfate complex, the ellagic acid-copper organic framework material, and the ellagic acid-copper-doxorubicin complex in Example 2 of the present invention;

[0026] Figure 6Fourier transform infrared spectra of the ellagic acid-copper-doxorubicin-chondroitin sulfate complex, the ellagic acid-copper organic framework material, and the ellagic acid-copper-doxorubicin complex in Example 2 of the present invention;

[0027] Figure 7 X-ray diffraction spectra of the ellagic acid-copper-doxorubicin-chondroitin sulfate complex, the ellagic acid-copper organic framework material, and the ellagic acid-copper-doxorubicin complex in Example 2 of the present invention;

[0028] Figure 8 This is a graph showing the in vitro stability test results of the ellagic acid-copper-doxorubicin-chondroitin sulfate complex in Example 2 of the present invention;

[0029] Fig. 9 The statistical graph of the survival rate of MCF-7 / Adr cells calculated by CCK-8 experiment after adding ellagic acid-copper-doxorubicin-chondroitin sulfate complex, ellagic acid-copper organic framework material, ellagic acid-copper-doxorubicin complex and doxorubicin at different concentrations to MCF-7 / Adr cells in Example 3 of the present invention;

[0030] Fig.10 The ellagic acid-copper-doxorubicin-chondroitin sulfate complex, ellagic acid-copper organic framework material, ellagic acid-copper-doxorubicin complex, and doxorubicin in Example 3 of the present invention are added to MCF-7 cells at different concentrations and then a CCK-8 experiment is performed to calculate the survival rate of MCF-7 cells;

[0031] Fig.11 A statistical graph of the survival rate of 4T1 cells calculated by CCK-8 experiment after adding ellagic acid-copper-doxorubicin-chondroitin sulfate complex, ellagic acid-copper organic framework material, ellagic acid-copper-doxorubicin complex and doxorubicin at different concentrations to 4T1 cells in Example 3 of the present invention;

[0032] Fig.12 The photographs of cell clone formation after the MCF-7 / Adr drug-resistant cells, MCF-7 cells, and 4T1 cells were subjected to cloning experiments after 5 mM saline, ellagic acid-copper-doxorubicin-chondroitin sulfate complex, ellagic acid-copper organic framework material, ellagic acid-copper-doxorubicin complex, and doxorubicin were applied respectively in Example 3 of the present invention;

[0033] Fig.13 This is a statistical graph of clone formation experiment after applying 5 mM physiological saline, ellagic acid-copper-doxorubicin-chondroitin sulfate complex, ellagic acid-copper organic framework material, ellagic acid-copper-doxorubicin complex, and doxorubicin to MCF-7 / Adr drug-resistant cells in Example 3 of the present invention and performing a clone experiment;

[0034] Fig.14This is a statistical graph of clone formation experiment after applying 5 mM physiological saline, ellagic acid-copper-doxorubicin-chondroitin sulfate complex, ellagic acid-copper organic framework material, ellagic acid-copper-doxorubicin complex, and doxorubicin to MCF-7 cells in Example 3 of the present invention and performing a clone experiment;

[0035] Fig.15 This is a statistical graph of clone formation experiment after applying 5 mM physiological saline, ellagic acid-copper-doxorubicin-chondroitin sulfate complex, ellagic acid-copper organic framework material, ellagic acid-copper-doxorubicin complex, and doxorubicin to 4T1 cells in Example 3 of the present invention and performing a clone experiment;

[0036] Fig.16 The distribution diagram of the relevant preparations in the mouse body and the enrichment diagram in the main organs after the mouse breast cancer in situ model was injected with normal saline, ellagic acid-copper-doxorubicin-chondroitin sulfate complex, ellagic acid-copper organic framework material, ellagic acid-copper-doxorubicin complex, and doxorubicin into the tail vein in Example 4 of the present invention;

[0037] Fig.17 This is a graph showing the experimental results of a tumor inhibition experiment conducted after normal saline, ellagic acid-copper-doxorubicin-chondroitin sulfate complex, ellagic acid-copper organic framework material, ellagic acid-copper-doxorubicin complex, and doxorubicin were injected into the tail vein of an in situ mouse breast cancer model in Example 4 of the present invention; DETAILED DESCRIPTION

[0038] In order to make the above-mentioned purposes, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below. It should be noted that the following embodiments are only used to illustrate the implementation method and typical parameters of the present invention, and are not used to limit the parameter range described in the present invention. Reasonable changes derived therefrom are still within the scope of protection of the claims of the present invention.

[0039] It should be noted that the endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be regarded as specifically disclosed in this article.

[0040] The occurrence and development of tumors is a complex process of multiple genes, multiple stages and multiple steps. It is an inseparable whole with the tumor microenvironment (TME). A large number of studies have shown that it helps to promote tumor proliferation, invasion, adhesion, and angiogenesis, thereby promoting the generation of malignant tumors. At the same time, it shows that low pH should be one of the important factors affecting tumor metastasis. Compared with normal cell tissues, tumor cells have a lower pH. The specific embodiment of the present invention provides a preparation method and application of a polyphenol metal coordination compound. This compound is based on ellagic acid-copper organic framework material. In the case of cooperating with doxorubicin and chondroitin sulfate, it can better kill tumor cells and enhance the sensitivity of doxorubicin-resistant cells. The principle is that ellagic acid is a natural compound with a polyphenol dilactone structure. Recent studies have reported that ellagic acid has a significant inhibitory effect on a variety of tumors, especially a variety of drug-resistant tumor cells. Ellagic acid has shown significant inhibition of cell survival-related pathways such as the AKT pathway and the MAPK pathway in a variety of tumors, and also has a significant downregulation effect on cell stemness-related pathways including Notch and Hedgehog. In addition, ellagic acid can also interfere with the energy supply in cells by destroying mitochondria, thereby affecting the drug efflux mediated by the ATP-dependent multidrug resistance receptor P-gp and reversing tumor resistance. Mitochondrial damage mediated by ellagic acid can also promote apoptosis of tumor cells, and can significantly enhance tumor killing when used in combination with chemotherapy drugs. The ellagic acid-copper organic framework material provided in the specific embodiment of the present invention is structurally stable under a neutral environment, but decomposes under the low pH environment of tumor cells. Therefore, the ellagic acid-copper organic framework material is released in tumor cells as a drug carrier, and doxorubicin can be embedded in the double strand of DNA to form a stable complex, affecting the function of DNA, preventing tumor DNA replication and RNA transcription; ellagic acid can mediate damage to tumor cell mitochondria and promote cell apoptosis; copper ions can mediate cell copper death, and doxorubicin has a strong anti-cancer effect. Chondroitin sulfate is highly expressed on the surface of tumors, so it can be used as a packaging material for active targeted delivery of tumors.

[0041] In a specific embodiment of the present invention, the steps for preparing the ellagic acid-copper-adriamycin-chondroitin sulfate complex include:

[0042] S1. Preparation of ellagic acid-copper organic framework material: adding ellagic acid to a sodium hydroxide solution with a concentration of 0.1-0.5 M, stirring, adding a copper sulfate solution with a copper ion concentration of 10-50 mM, the molar ratio of ellagic acid to copper ion being 1:(0.5-2), stirring, centrifuging, and lyophilizing the supernatant to obtain the ellagic acid-copper organic framework material;

[0043] S2. Preparation of ellagic acid-copper-doxorubicin complex: Mix the ellagic acid-copper organic framework material prepared in step S1 with doxorubicin, wherein the mass ratio of the ellagic acid-copper organic framework material to doxorubicin is 1:(0.5-2), add water to dissolve, stir and centrifuge, collect the precipitate, wash with water and freeze-dry to obtain the ellagic acid-copper-doxorubicin complex;

[0044] S3. Preparation of ellagic acid-copper-doxorubicin-chondroitin sulfate complex: the ellagic acid-copper-doxorubicin complex of step S2 is dispersed in a chondroitin sulfate aqueous solution with a concentration of 1-5 mg / mL, and the mass ratio of chondroitin sulfate to ellagic acid-copper-doxorubicin complex is 1:(5-10). After stirring, centrifugation is performed, and the precipitate is collected, washed with water, and then freeze-dried to obtain the ellagic acid-copper-doxorubicin-chondroitin sulfate complex.

[0045] To make the embodiments of the present invention easy to understand, in the following embodiments, ellagic acid is abbreviated as EA, ellagic acid-copper organic framework material will be abbreviated as EA-CU, doxorubicin is abbreviated as DOX, ellagic acid-copper-doxorubicin complex is abbreviated as EC@DOX or NPs, chondroitin sulfate is abbreviated as CS, and ellagic acid-copper-doxorubicin-chondroitin sulfate complex is abbreviated as CS / EC@DOX or CS / NPs.

[0046] Example 1

[0047] Preparation of CS / EC@DOX. The specific preparation process is as follows Figure 1 Shown

[0048] S1: Preparation of EA-CU: Weigh 0.2 g NaOH and add it to 20 mL deionized water to prepare 0.25 M NaOH solution, weigh 168 mg CuSO4·5H2O solid and add it to 20 mL deionized water to prepare 33.6 mM Cu 2+ Weigh 100 mg of ellagic acid (EA) and add 20 mL of 0.25 M NaOH solution. Stir at room temperature for 24 hours at 1000 rpm. Then add 33.6 mM Cu 2+ (Molar ratio: EA:Cu 2+ =1:1), stirred at 1000 rpm for 24 hours at room temperature, and then centrifuged (10000 rpm, 15 min). The supernatant was placed in a glass dish and frozen in a -80° refrigerator overnight. It was then placed in a freeze dryer for 48 hours. The freeze-dried powder was retained to obtain EA-CU.

[0049] S2. Preparation of EC@DOX: EA-Cu synthesized in step S1 was mixed with DOX in a mass ratio of 1:1, dissolved in deionized water, stirred at 1000 rpm at room temperature for 24 hours, and then centrifuged (10000 rpm, 15 min). The precipitate was collected, washed three times with water, and the retained precipitate was freeze-dried to obtain EC@DOX.

[0050] S3, preparation of CS / EC@DOX: The EC@DOX powder synthesized in step S2 was dispersed in a chondroitin sulfate (CS) aqueous solution, stirred at 1000 rpm for 24 hours at room temperature, centrifuged at 10000 rpm for 15 minutes, the precipitate was collected, washed with water three times, and the retained precipitate was freeze-dried to obtain CS / EC@DOX. Chondroitin sulfate (CS) was prepared in a 3 mg / mL deionized water solution.

[0051] Example 2

[0052] The particle sizes of EA-Cu, EC@DOX, and CS / EC@DOX prepared in Example 1 were measured by dynamic light scattering, and the morphology of the preparations was photographed by TEM. The Zeta potential, in vitro stability, and Fourier transform infrared spectra of the preparations were measured. The final results are as follows: Figure 2~Figure 8 shown.

[0053] in Figure 2 The particle size of EA-CU and its electron microscope photo; Figure 3 The particle size of EC@DOX and its photo under electron microscope; Figure 4 The particle size of CS / EC@DOX and its electron microscope photo; Figure 5 Zeta potential measurement results of EA-Cu, EC@DOX, and CS / EC@DOX; Figure 6 Fourier transform infrared spectra of EA-Cu, EC@DOX, and CS / EC@DOX; Figure 7 X-ray diffraction results of EA-Cu, EC@DOX, and CS / EC@DOX; Figure 8 Color change of CS / EC@DOX after being placed at room temperature for 7 days

[0054] Depend on Figure 2~Figure 8 It can be seen that EA-Cu, EC@DOX, and CS / EC@DOX are all nanoparticles with diameters ranging from 80 to 200 nm, and the absolute value of the Zeta potential of CS / EC@DOX is much higher than that of EA-Cu and EC@DOX, which means that CS / EC@DOX has good stability. Figure 8 Proved this.

[0055] Example 3

[0056] Different concentrations of DOX, EA-CU, EC@DOX, and CS / EC@DOX were used to treat different cells at different time points to verify the auxiliary effects of EA-CU and CS on drug uptake; CCK8 cytotoxicity assay and clone formation assay were used to verify the proliferation inhibition and killing effects of DOX, EA-CU, EC@DOX, and CS / EC@DOX on tumor MCF-7 and MCF-7 / Adr drug-resistant cells in vitro, as well as the synergistic killing effect of EA-CU and DOX after nanoformulation packaging. The experimental results are shown in Figure 2. Figures 9 to 15 shown.

[0057] Fig. 9 The survival rate of MCF-7 / Adr drug-resistant cells was calculated by CCK-8 experiment after treating MCF-7 / Adr drug-resistant cells with different concentrations of DOX, EA-CU, EC@DOX, and CS / EC@DOX. Fig. 9 It can be seen that EA-CU has similar anti-cancer properties to DOX for drug-resistant cells. At the same time, at high concentrations (molar concentration greater than or equal to 5 μM), the anti-cancer properties of EC@DOX and CS / EC@DOX for drug-resistant cells are significantly better than those of DOX and EA-CU. CS / EC@DOX can significantly enhance the tumor killing ability of DOX. This experiment indirectly proves that CS / EC@DOX has the effect of reversing doxorubicin resistance in breast cancer.

[0058] Fig.10 The survival rate of MCF-7 cells was calculated by CCK-8 experiment after treating MCF-7 cells with different concentrations of DOX, EA-CU, EC@DOX, and CS / EC@DOX. Fig.10 It can be seen that for common cancer cells, the tumor inhibition performance of DOX is slightly better than that of EA-CU, but the tumor inhibition performance of EC@DOX and CS / EC@DOX is significantly better than that of DOX and EA-CU. CS / EC@DOX has the best tumor inhibition performance. CS / EC@DOX can significantly enhance the tumor killing ability of DOX.

[0059] Fig.11 The survival rate of 4T1 cells was calculated by CCK-8 experiment after 4T1 cells were treated with different concentrations of DOX, EA-CU, EC@DOX, and CS / EC@DOX. This experiment proves that Fig.11 The conclusions of the experiments shown also indicate that EA-CU, EC@DOX, and CS / EC@DOX all have significant anti-cancer properties for different breast cancer cells from different sources, and CS / EC@DOX can significantly enhance the tumor killing ability of DOX.

[0060] Fig.12The following are photos of cell clone formation after applying 5 mM saline, DOX, EA-CU, EC@DOX, and CS / EC@DOX to MCF-7 / Adr resistant cells, MCF-7 cells, and 4T1 cells, respectively. Figure 13~Figure 15 From the statistical graph of clone formation, it can be concluded that both EC@DOX and CS / EC@DOX have good anti-growth ability against tumor cells, among which CS / EC@DOX has the best anti-growth ability against tumor cells.

[0061] Example 4

[0062] In vivo distribution assay was used to evaluate the in vivo distribution and tumor targeting abilities of DOX, EA-CU, EC@DOX, and CS / EC@DOX:

[0063] A luciferase reporter gene system was constructed using MCF-7 wild-type and MCF-7 drug-resistant cells, and an orthotopic mouse breast cancer model was constructed. The growth of the orthotopic tumors in mice was monitored by fluorescence after tail vein injection of DOX, EC@DOX, CS / EC@DOX and saline (control group). The anti-tumor effects of DOX, EC@DOX, CS / EC@DOX and the effect of reversing doxorubicin resistance were verified in vivo. The experimental results are shown in Figure 2. Fig.16 , Fig.17 shown.

[0064] Fig.16 A in the figure shows the distribution of related preparations in mice 1h, 2h, 4h, 6h, 8h, and 24h after tail vein injection of DOX, EC@DOX, and CS / EC@DOX into the mouse breast cancer orthotopic model. Fig.16 Figure B shows the enrichment of related agents in the main organs of mice 24 hours after the tail vein injection of DOX, EC@DOX, and CS / EC@DOX into the mouse breast cancer orthotopic model. Fig.16 It can be seen that compared with DOX, EC@DOX and CS / EC@DOX have better inhibitory effects on tumors in vivo. After the three preparations are injected into the body, the main metabolic sites are located in the liver and kidneys. Among them, CS / EC@DOX can act precisely on tumor tissue as a targeted drug to exert an anti-tumor effect. At the same time, since this example uses a mouse doxorubicin-resistant model, this example can prove that CS / EC@DOX has the effect of reversing the doxorubicin resistance of breast cancer cells.

[0065] Fig.17 These are the experimental results of tumor inhibition experiments after injecting normal saline, DOX, EC@DOX, and CS / EC@DOX into the tail vein of an orthotopic mouse breast cancer model. Fig.17A in the figure is the measurement diagram of the tumor size of each group of mice after the tumor inhibition experiment. Fig.17 B in the figure is the curve of tumor mass and inhibition rate in each group. Fig.17 It can be seen that CS / EC@DOX has a significant inhibitory effect on the tumor of the mouse breast cancer in situ model. At the same time, since this example uses a mouse doxorubicin-resistant model, this example can prove that CS / EC@DOX has the effect of reversing the doxorubicin resistance of breast cancer cells.

[0066] The mechanism of action of the ellagic acid-copper-doxorubicin-chondroitin sulfate complex in a specific embodiment of the present invention is as follows: the ellagic acid-copper-doxorubicin-chondroitin sulfate complex accurately reaches the tumor cells under the action of chondroitin sulfate and enters the tumor cells by endocytosis. In the low pH intracellular environment of the tumor cells, the ellagic acid-copper-doxorubicin-chondroitin sulfate complex decomposes to produce doxorubicin. In the tumor cells, doxorubicin accumulates to kill the tumor. At the same time, the ellagic acid produced by the decomposition of the ellagic acid-copper-doxorubicin-chondroitin sulfate complex can mediate the functional disorder of the mitochondria of the tumor cells, induce mitochondrial apoptosis to kill the tumor cells, and further, the copper ions produced by the decomposition of ellagic acid-copper-doxorubicin-chondroitin sulfate can mediate the copper death pathway of the tumor cells. For tumor cells resistant to doxorubicin, doxorubicin will be pumped out of the tumor cells through the P-gp glycoprotein, but the mitochondrial apoptosis and copper death pathways can still play a role, which will increase the sensitivity of doxorubicin-resistant cells to doxorubicin and achieve the purpose of reversing the resistance of doxorubicin-resistant cells.

[0067] Although the disclosure is disclosed as above, the protection scope of the disclosure is not limited thereto. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the disclosure, and these changes and modifications will fall within the protection scope of the present invention.

Claims

1. A method for preparing a polyphenol metal coordination compound, characterized in that: The preparation method comprises the following steps: S1: adding ellagic acid to a sodium hydroxide solution, stirring, adding a copper ion solution, stirring, centrifuging, and lyophilizing the supernatant to obtain an ellagic acid-copper organic framework material; S2: mixing ellagic acid-copper organic framework material and doxorubicin, adding water to dissolve, stirring and centrifuging, collecting precipitates, washing with water and freeze-drying to obtain ellagic acid-copper-doxorubicin complex; S3: dispersing the ellagic acid-copper-doxorubicin complex in the chondroitin sulfate aqueous solution, stirring and centrifuging, collecting the precipitate, washing with water and freeze-drying to obtain the ellagic acid-copper-doxorubicin-chondroitin sulfate complex.

2. The method for preparing a polyphenol metal coordination compound according to claim 1, characterized in that: The concentration of the sodium hydroxide solution in step S1 is 0.1-0.5 M.

3. The method for preparing a polyphenol metal coordination compound according to claim 1, characterized in that: The copper ion solution in step S1 is a copper sulfate solution and / or a copper chloride solution.

4. The method for preparing a polyphenol metal coordination compound according to claim 3, characterized in that: The copper ion concentration in the copper ion solution is 10-50 mM.

5. The method for preparing a polyphenol metal coordination compound according to claim 1, characterized in that: In step S1, the molar ratio of ellagic acid to copper ions is 1:(0.5-2).

6. The method for preparing a polyphenol metal coordination compound according to claim 1, characterized in that: In the step S2, the mass ratio of ellagic acid-copper organic framework material to doxorubicin is 1:(0.5-2).

7. The method for preparing a polyphenol metal coordination compound according to claim 1, characterized in that: The concentration of the chondroitin sulfate aqueous solution in step S3 is 1-5 mg / mL.

8. The method for preparing a polyphenol metal coordination compound according to claim 1, characterized in that: In the step S3, the mass ratio of chondroitin sulfate to ellagic acid-copper-adriamycin complex is 1:(5-10).

9. A polyphenol metal coordination compound, characterized in that: The polyphenol metal coordination compound is prepared by any preparation method according to claims 1 to 8.

10. An application of the polyphenol metal coordination compound according to claim 9, characterized in that: The polyphenol metal coordination compound is applied to the preparation of drugs for treating breast cancer.

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