A metal polyphenol nanoparticle, its preparation method, and its application in the preparation of TMEM16A and / or EGFR inhibitors.

Abstract

This invention provides metal polyphenol nanoparticles, a preparation method, and their application in the preparation of TMEM16A and / or EGFR inhibitors. The metal polyphenol nanoparticles comprise an organic framework formed by the reaction of proanthocyanidins and PEG polyphenol compounds, and metal nanoparticles modified by metal coordination within the organic framework. This invention also provides metal polyphenol nanoparticles containing an anti-breast cancer drug, comprising: the aforementioned metal polyphenol nanoparticles, and an EGFR inhibitor drug coated within the metal polyphenol nanoparticle network. Furthermore, both types of metal polyphenol nanoparticles also include a hyaluronic acid shell. The metal polyphenol nanoparticles of this invention establish a dual-target nanonetwork of TMEM16A and EGFR, which can synergistically inhibit breast cancer proliferation, migration, and recurrence, thereby achieving efficient and specific treatment of breast cancer.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a metal polyphenol nanoparticle, its preparation method, and its application in the preparation of TMEM16A and / or EGFR inhibitors. Background Technology

[0002] Breast cancer is a very common cancer among women, with approximately 2 million new cases worldwide each year. Both incidence and mortality rates are on the rise, and the disease burden is increasing, making it a major global public health issue. Triple-negative breast cancer is the most aggressive subtype of breast cancer, accounting for about 15% of all breast cancers. Currently, clinical treatment options are very limited. Compared with other subtypes of breast cancer, advanced triple-negative breast cancer is characterized by high invasiveness, high early recurrence rate, high rate of visceral metastasis, and poor prognosis. Advanced triple-negative breast cancer is not sensitive to targeted therapy and endocrine therapy, lacking specific treatment methods; traditional treatments mainly consist of surgery, radiotherapy, and chemotherapy.

[0003] With the development of medicine, the treatment methods for breast cancer patients have become more and more perfect. However, the treatment process is accompanied by corresponding side effects. Since triple-negative breast cancer lacks effective treatment targets, its main treatment methods are surgical resection and radiotherapy and chemotherapy.

[0004] Surgical treatment can achieve maximum local control of the primary tumor and regional lymph nodes. However, as breast cancer progresses to the mid-stage, patients are often weak and metastasis is highly likely. Blindly performing surgery can significantly weaken the patient, making recovery difficult and increasing the risk of recurrence, which is even more challenging to treat. Chemotherapy, on the other hand, is extremely harmful to the body. While chemotherapy drugs kill cancer cells, they also kill normal cells, leading to complications such as decreased white blood cell count, bone marrow suppression, nausea, vomiting, and weight loss. Furthermore, due to drug resistance, both single-agent and combination chemotherapy are less effective, with a median survival of 9-12 months and a 5-year survival rate of less than 30%. Chemotherapy resistance has become a bottleneck in the treatment of triple-negative breast cancer. Therefore, developing effective treatment options for breast cancer is urgently needed.

[0005] At the same time, an increasing number of nanomedicines have provided new ideas for the treatment of breast cancer. However, most conventional nanomedicine delivery systems require the synthesis of additional carriers and the synthesis routes are relatively complex. These processes are difficult to control precisely in space and time, have poor reproducibility, and reduce the efficiency of drug treatment. In addition, most carriers have poor biocompatibility and are difficult to metabolize and excrete, thus reducing their effectiveness in in vitro and in vivo experiments.

[0006] In view of this, the present invention is hereby proposed. Summary of the Invention

[0007] One objective of this invention is to provide a metal polyphenol nanoparticle, comprising an organic framework formed by the reaction of proanthocyanidins and PEG polyphenol compounds, and metal nanoparticles modified by metal coordination within the organic framework. The metal polyphenol nanoparticle uses the natural polyphenol small molecule inhibitor proanthocyanidins as its framework, with metal ions coordinating to form a metal polyphenol nanonetwork. This network can efficiently encapsulate anti-breast cancer drugs, such as the EGFR inhibitor gefitinib, and utilizes the photothermal properties of the metal polyphenol nanonetwork to achieve chemotherapy-PTT synergistic therapy.

[0008] A second objective of this invention is to provide a method for preparing the aforementioned metal polyphenol nanoparticles. The method includes the following steps: mixing a metal salt solution and a PEG polyphenol compound solution, followed by a first stirring; then mixing with a proanthocyanidin solution, followed by a second stirring; and finally collecting the metal polyphenol nanoparticles after the reaction is complete. The method for preparing the metal polyphenol nanoparticles of this invention is simple and easy to operate, and can produce nanoparticles with uniform size and spherical morphology.

[0009] A third objective of this invention is to provide metal polyphenol nanoparticles containing anti-breast cancer drugs. These nanoparticles comprise the aforementioned metal polyphenol nanoparticles and the anti-breast cancer drug encapsulated within a network of these nanoparticles. This polyphenol nanonetwork efficiently encapsulates anti-breast cancer drugs, such as the EGFR inhibitor gefitinib, forming a dual-target nanonetwork of TMEM16A and EGFR. The dual targets synergistically inhibit breast cancer proliferation, migration, and recurrence, thereby achieving efficient and specific treatment of breast cancer.

[0010] The fourth objective of this invention is to provide a method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs. The preparation method includes the following steps: mixing a metal salt solution and a PEG polyphenol compound solution, followed by a first stirring; then mixing with a proanthocyanidin solution, followed by a second stirring; then mixing with a gefitinib solution, followed by a third stirring; after the reaction is complete, collecting the metal polyphenol nanoparticles containing the anti-breast cancer drug.

[0011] The fifth objective of this invention is to provide the application of metal polyphenol nanoparticles, or metal polyphenol nanoparticles containing anti-breast cancer drugs, in the preparation of TMEM16A and / or EGFR inhibitors.

[0012] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted:

[0013] In a first aspect, the present invention provides metal polyphenol nanoparticles, the metal polyphenol nanoparticles comprising an organic framework body formed by the reaction of proanthocyanidins and PEG polyphenol compounds, and metal nanoparticles modified in the organic framework body by metal coordination.

[0014] This invention utilizes proanthocyanidins, a small-molecule inhibitor of natural polyphenols, as a framework, with metal ions coordinating to form a metal-polyphenol nanonetwork. Compared to traditional nanoparticles, metal-polyphenol nanoparticles with proanthocyanidins as the framework do not require the introduction of an additional carrier; they can form a nanonetwork simply by coordinating with metals. Furthermore, the metal-polyphenol nanoparticles of this invention possess excellent photothermal responsiveness, rapidly heating up intracellularly or extracellularly under near-infrared light irradiation. They can also efficiently encapsulate anti-breast cancer drugs, such as the EGFR inhibitor gefitinib. The photothermal properties of the metal-polyphenol nanonetwork enable synergistic chemotherapy-PTT therapy.

[0015] Among them, procyanidins, also known as proanthocyanidins, are polyphenolic compounds that can produce anthocyanins under hot acid treatment. They are widely distributed and rich in variety, mainly found in natural plants such as grape seeds, ginkgo leaves, lotus seedpods, Japanese cypress, and French coastal pine bark. They have antioxidant and free radical scavenging abilities. At the same time, procyanidins are also a highly effective natural small molecule TMEM16A channel inhibitor with excellent anti-tumor activity.

[0016] Preferably, the PEG polyphenol compound is obtained by reacting an eight-arm polyethylene glycol activated ester with a dopamine salt.

[0017] Preferably, the molecular weight of the eight-arm polyethylene glycol activated ester is 8000-12000 Da, for example, it can be 8000 Da, 8500 Da, 9000 Da, 9500 Da, 10000 Da, 10500 Da, 11000 Da, 11500 Da, 12000 Da, etc.

[0018] Preferably, the eight-arm polyethylene glycol activated ester is an eight-arm polyethylene glycol succinimide ester.

[0019] Preferably, the dopamine salt is dopamine hydrochloride.

[0020] Preferably, the metal nanoparticles are selected from any one or a combination of at least two of Cu, Zr, or Fe, with Fe being the most preferred.

[0021] Preferably, the metal polyphenol nanoparticles further include a hyaluronic acid shell.

[0022] In this invention, the outer layer of the nanonetwork is coated with a layer of hyaluronic acid (HA) as a shell, which can target the CD44 receptor in tumor cells, providing a new solution for the research and development of targeted therapies for breast cancer.

[0023] Preferably, the molecular weight of the hyaluronic acid is 8000-12000 Da, for example, it can be 8000 Da, 8500 Da, 9000 Da, 9500 Da, 10000 Da, 10500 Da, 11000 Da, 11500 Da, 12000 Da, etc.

[0024] In a second aspect, the present invention provides a method for preparing metal polyphenol nanoparticles as described in the first aspect, the method comprising the following steps:

[0025] The metal salt solution and the PEG polyphenol compound solution were mixed and stirred for the first time; then mixed with the proanthocyanidin solution and stirred for the second time; after the reaction was completed, the metal polyphenol nanoparticles were collected.

[0026] This method enables the simple and efficient preparation of metal polyphenol nanoparticles based on polyphenol TMEM16A inhibitors, which can achieve local treatment of breast cancer by inhibiting the overexpression of TMEM16A channels. This method is of great significance for the research and development, quality control and even clinical application of targeted therapies for breast cancer.

[0027] Preferably, the metal polyphenol nanoparticles are PF-NPs nanoparticles.

[0028] Preferably, the PEG polyphenol compound is prepared by the following steps:

[0029] First, an eight-arm polyethylene glycol activated ester, dopamine salt, and solvent are mixed and reacted; then, an acid-binding agent is added to the above reaction system and the reaction is carried out; after the reaction is completed, the PEG polyphenol compound is collected.

[0030] Preferably, the mass ratio of the eight-arm polyethylene glycol activated ester to the dopamine salt is 10:(3-5), for example, it can be 10:3, 10:3.2, 10:3.4, 10:3.6, 10:3.8, 10:4, 10:4.2, 10:4.4, 10:4.6, 10:4.8, 10:5, etc.

[0031] Preferably, the solvent is N,N-dimethylformamide (DMF).

[0032] Preferably, the acid-binding agent is triethylamine (TEA, Et3N).

[0033] Preferably, the amount of acid-binding agent added is 0.1% to 1% of the total mass of the eight-arm polyethylene glycol activated ester and dopamine salt, for example, it can be 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, etc.

[0034] Preferably, before adding the acid-binding agent to the above reaction system, the reaction is carried out at 20-30°C (e.g., 20°C, 22°C, 24°C, 26°C, 28°C, 30°C, etc.) for 0.5-2 hours, for example, 0.5 hours, 0.6 hours, 0.8 hours, 1 hour, 1.2 hours, 1.4 hours, 1.6 hours, 1.8 hours, 2 hours, etc.

[0035] Preferably, after adding the acid-binding agent to the above reaction system, the reaction is carried out at 20-30°C (e.g., 20°C, 22°C, 24°C, 26°C, 28°C, 30°C, etc.) for 10-15 hours, for example, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, etc.

[0036] Preferably, after the reaction is completed, the collected product needs to be dialyzed under slightly acidic conditions.

[0037] Preferably, the slightly acidic condition is an aqueous solution of hydrochloric acid.

[0038] Preferably, the concentration of the hydrochloric acid aqueous solution is 0.5 to 2 mol / L, for example, it can be 0.5 mol / L, 0.6 mol / L, 0.8 mol / L, 1 mol / L, 1.2 mol / L, 1.4 mol / L, 1.6 mol / L, 1.8 mol / L, 2 mol / L, etc.

[0039] Preferably, the dialysis time is 42 to 54 hours, for example, 42 hours, 44 hours, 46 hours, 48 ​​hours, 50 hours, 52 hours, 54 hours, etc.

[0040] Preferably, the dialysis process further includes lyophilization, wherein the lyophilization temperature is -80 to -40°C, for example, -80°C, -75°C, -70°C, -65°C, -60°C, -55°C, -50°C, -45°C, -40°C, etc., and the lyophilization time is 24 to 48 hours, for example, 24 hours, 30 hours, 36 hours, 42 hours, 48 ​​hours, etc.

[0041] As a preferred embodiment of the present invention, the PEG polyphenol compound is prepared by the following steps:

[0042] First, an eight-arm polyethylene glycol activated ester, dopamine salt, and solvent are mixed and reacted at 20–30°C for 0.5–2 h in the presence of a protective gas. Then, an acid-binding agent is added to the above reaction system, and the reaction is continued at 20–30°C for 10–15 h in the presence of a protective gas and in the dark. The product after the reaction is completed is dialyzed and lyophilized to obtain the PEG polyphenol compound.

[0043] Preferably, the mass ratio of the metal salt, PEG polyphenol compound, and proanthocyanidins is (4-6):(5-7):(2-3);

[0044] Among them, "4 to 6" can be, for example, 4, 4.2, 4.4, 4.6, 4.8, 5, 5.2, 5.4, 5.6, 5.8, 6, etc.;

[0045] Among them, "5 to 7" can be, for example, 5, 5.2, 5.4, 5.6, 5.8, 5, 6.2, 6.4, 6.6, 6.8, 7, etc.;

[0046] Among them, "2 to 3" can be, for example, 2, 2.2, 2.4, 2.6, 2.8, 6, etc.

[0047] Preferably, the concentration of the metal salt solution is 50-150 mg / mL, for example, it can be 50 mg / mL, 60 mg / mL, 80 mg / mL, 100 mg / mL, 120 mg / mL, 140 mg / mL, 150 mg / mL, etc.

[0048] Preferably, the concentration of the PEG polyphenol compound solution is 1 to 5 mg / mL, for example, it can be 1 mg / mL, 1.5 mg / mL, 2 mg / mL, 2.5 mg / mL, 3 mg / mL, 3.5 mg / mL, 4 mg / mL, 4.5 mg / mL, 5 mg / mL, etc.

[0049] Preferably, the concentration of the proanthocyanidin solution is 5–15 mg / mL, for example, it can be 5 mg / mL, 6 mg / mL, 8 mg / mL, 10 mg / mL, 12 mg / mL, 14 mg / mL, 15 mg / mL, etc.

[0050] Preferably, the temperature of the first stirring is 20-30℃, for example, 20℃, 22℃, 24℃, 26℃, 28℃, 30℃, etc., and the time is 0.5-2h, for example, 0.5h, 0.6h, 0.7h, 0.8h, 0.9h, 1h, 1.2h, 1.4h, 1.6h, 1.8h, 2h, etc.

[0051] Preferably, the temperature of the second stirring is 20-30℃, for example, 20℃, 22℃, 24℃, 26℃, 28℃, 30℃, etc., and the time is 6-8h, for example, 6h, 6.2h, 6.5h, 6.8h, 7h, 7.2h, 7.5h, 7.8h, 8h, etc.

[0052] Preferably, the method for collecting the metal polyphenol nanoparticles is centrifugation, wherein the centrifugation speed is 8000-12000 r / min, for example, 8000 r / min, 9000 r / min, 10000 r / min, 11000 r / min, 12000 r / min, etc., and the centrifugation time is 5-15 min, for example, 5 min, 6 min, 8 min, 10 min, 11 min, 12 min, 13 min, 14 min, 15 min, etc.

[0053] Preferably, the centrifugation process further includes washing, which is washing with water at least once, for example, once, twice, three times, four times, five times, etc.

[0054] Preferably, the product obtained from washing needs to be redispersed in water for storage, and the mass ratio of the product to water is 1:(3-5), for example, it can be 1:3, 1:3.2, 1:3.4, 1:3.6, 1:3.8, 1:4, 1:4.2, 1:4.4, 1:4.6, 1:4.8, 1:5, etc.

[0055] Preferably, the preparation method of the metal polyphenol nanoparticles further includes coating with a hyaluronic acid shell, specifically comprising the following steps:

[0056] Metal polyphenol nanoparticles were placed in a hyaluronic acid solution, coated by stirring, and then centrifuged to obtain hyaluronic acid-coated metal polyphenol nanoparticles.

[0057] Preferably, the preparation method of the metal polyphenol nanoparticles further includes coating with a hyaluronic acid shell, specifically comprising the following steps:

[0058] PF-NPs nanoparticles were placed in a hyaluronic acid solution, coated by stirring, and then centrifuged to obtain HA-PF-NPs nanoparticles.

[0059] Preferably, the mass ratio of the PF-NPs nanoparticles to the hyaluronic acid solution is 1:(0.5~1), for example, it can be 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, etc.

[0060] Preferably, the concentration of the hyaluronic acid solution is 0.5 to 1 mg / mL, for example, it can be 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1 mg / mL, etc.

[0061] Preferably, the stirring is carried out under light-protected conditions.

[0062] Preferably, the stirring temperature is 20-30℃, for example, 20℃, 22℃, 24℃, 26℃, 28℃, 30℃, etc., and the stirring time is 6-8h, for example, 6h, 6.2h, 6.4h, 6.6h, 6.8h, 7h, 7.2h, 7.4h, 7.6h, 7.8h, 8h, etc.

[0063] As a preferred embodiment of the present invention, the metal polyphenol nanoparticles are prepared by the following steps:

[0064] Preparation of S1 and PEG polyphenol compounds

[0065] First, an eight-arm polyethylene glycol activated ester, dopamine salt, and solvent are mixed and reacted at 20–30°C for 0.5–2 h in the presence of a protective gas. Then, an acid-binding agent is added to the above reaction system, and the reaction continues at 20–30°C for 10–15 h in the presence of a protective gas and in the dark. The product after the reaction is completed is dialyzed and lyophilized to obtain the PEG polyphenol compound.

[0066] S2. Preparation of metal polyphenol nanoparticles

[0067] The metal salt solution and the PEG polyphenol compound solution were mixed and stirred at 20–30°C for 0.5–1 h; then, the proanthocyanidin solution was added to the above system and the mixture was stirred at 20–30°C for 4–8 h; after the reaction was completed, the mixture was centrifuged and washed to obtain the metal polyphenol nanoparticles.

[0068] S3, a transparent outer shell covering

[0069] Metal polyphenol nanoparticles were placed in a hyaluronic acid solution and stirred at 20–30°C for 6–8 hours to complete the coating. Then, they were centrifuged to obtain metal polyphenol nanoparticles coated with a transparent shell.

[0070] Thirdly, the present invention provides a metal polyphenol nanoparticle containing an anti-breast cancer drug, wherein the metal polyphenol nanoparticle containing the anti-breast cancer drug comprises: the metal polyphenol nanoparticle described in the first aspect, and the anti-breast cancer drug coated in the metal polyphenol nanoparticle network.

[0071] Preferably, the anti-breast cancer drug is an EGFR inhibitor.

[0072] Preferably, the EGFR inhibitor is gefitinib.

[0073] Gefitinib is a first-generation EGFR inhibitor (a small molecule compound). Inhibition of EGFR can hinder tumor growth, metastasis, and angiogenesis, and increase tumor cell apoptosis. This invention combines gefitinib with a metallopolyphenol nanonetwork to achieve synergistic chemotherapy-PTT therapy. This approach overcomes both gefitinib resistance and the susceptibility of anthocyanins to oxidation, providing a highly effective solution for breast cancer treatment through their synergistic action on two targets.

[0074] Preferably, the loading of the EGFR inhibitor is 3 to 8 wt%, for example, it can be 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, 5 wt%, 5.5 wt%, 6 wt%, 6.5 wt%, 7 wt%, 7.5 wt%, 8 wt%, etc.

[0075] Preferably, the metal polyphenol nanoparticles containing anti-breast cancer drugs further include a hyaluronic acid shell.

[0076] In this invention, a layer of hyaluronic acid (HA) is coated on the outer layer of a metal polyphenol nanonetwork containing anti-breast cancer drugs as a shell, which can target the CD44 receptor in tumor cells, providing a new solution for the research and development of targeted therapies for breast cancer.

[0077] Preferably, the molecular weight of the hyaluronic acid is 8000-12000 Da, for example, it can be 8000 Da, 8500 Da, 9000 Da, 9500 Da, 10000 Da, 10500 Da, 11000 Da, 11500 Da, 12000 Da, etc.

[0078] Fourthly, the present invention provides a method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs as described in the third aspect, the preparation method comprising the following steps:

[0079] The metal salt solution and the PEG polyphenol compound solution were mixed and stirred for the first time; then mixed with the proanthocyanidin solution and stirred for the second time; then mixed with the gefitinib solution and stirred for the third time; after the reaction was completed, the metal polyphenol nanoparticles containing the anti-breast cancer drug were collected.

[0080] Preferably, the metal polyphenol nanoparticles containing anti-breast cancer drugs are PFG-NPs nanoparticles.

[0081] Preferably, the concentration of the gefitinib solution is 5–15 mg / mL, for example, it can be 5 mg / mL, 6 mg / mL, 8 mg / mL, 10 mg / mL, 12 mg / mL, 14 mg / mL, 15 mg / mL, etc.

[0082] Preferably, the amount of gefitinib solution added accounts for 2 to 6% of the total volume of the mixed solution, for example, it can be 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, etc.

[0083] Preferably, the temperature of the first stirring is 20-30℃, for example, 20℃, 22℃, 24℃, 26℃, 28℃, 30℃, etc., and the time is 0.5-1h, for example, 0.5h, 0.6h, 0.7h, 0.8h, 0.9h, 1h, etc.

[0084] Preferably, the temperature of the second stirring is 20-30°C, for example, 20°C, 22°C, 24°C, 26°C, 28°C, 30°C, etc., and the time is 5-10 minutes, for example, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, etc.

[0085] Preferably, the temperature of the third stirring is 20-30°C, for example, 20°C, 22°C, 24°C, 26°C, 28°C, 30°C, etc., and the time is 4-8 hours, for example, 4 hours, 4.5 hours, 5 hours, 5.5 hours, 6 hours, 6.5 hours, 7 hours, 7.5 hours, 8 hours, etc.

[0086] Preferably, the method for collecting the metal polyphenol nanoparticles containing anti-breast cancer drugs is centrifugation. The centrifugation speed is 8000-12000 r / min, for example, 8000 r / min, 9000 r / min, 10000 r / min, 11000 r / min, 12000 r / min, etc., and the centrifugation time is 10-15 min, for example, 10 min, 11 min, 12 min, 13 min, 14 min, 15 min, etc.

[0087] Preferably, the centrifugation process further includes washing, which is washing with water at least once, for example, once, twice, three times, four times, five times, etc.

[0088] Preferably, the product obtained from washing needs to be redispersed in water for storage, and the mass ratio of the product to water is 1:(0.8~1.2), for example, it can be 1:0.8, 1:0.85, 1:0.9, 1:0.95, 1:1, 1:1.05, 1:1.1, 1:1.15, 1:1.2, etc.

[0089] Preferably, the preparation method of the metal polyphenol nanoparticles further includes coating with a hyaluronic acid shell, specifically comprising the following steps:

[0090] Metal polyphenol nanoparticles containing anti-breast cancer drugs were placed in a hyaluronic acid solution, and the coating was completed by stirring. Then, the nanoparticles were centrifuged to obtain hyaluronic acid-coated metal polyphenol nanoparticles containing anti-breast cancer drugs.

[0091] Preferably, the preparation method of the metal polyphenol nanoparticles containing anti-breast cancer drugs further includes coating with a hyaluronic acid shell, specifically including the following steps:

[0092] PFG-NPs nanoparticles were placed in a hyaluronic acid solution, coated by stirring, and then centrifuged to obtain HA-PFG-NPs nanoparticles.

[0093] Preferably, the mass ratio of the PFG-NPs nanoparticles to the hyaluronic acid solution is 1:(0.5-1), for example, it can be 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, etc.

[0094] Preferably, the concentration of the hyaluronic acid solution is 0.5 to 1 mg / mL, for example, it can be 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1 mg / mL, etc.

[0095] Preferably, the stirring is carried out under light-protected conditions.

[0096] Preferably, the stirring temperature is 20-30℃, for example, 20℃, 22℃, 24℃, 26℃, 28℃, 30℃, etc., and the stirring time is 6-8h, for example, 6h, 6.2h, 6.4h, 6.6h, 6.8h, 7h, 7.2h, 7.4h, 7.6h, 7.8h, 8h, etc.

[0097] As a preferred technical solution of the present invention, the metal polyphenol nanoparticles containing anti-breast cancer drugs are prepared by the following steps (e.g. Figure 1 As shown):

[0098] Preparation of S1 and PEG polyphenol compounds

[0099] First, an eight-arm polyethylene glycol activated ester, dopamine salt, and solvent are mixed and reacted at 20–30°C for 0.5–2 h in the presence of a protective gas. Then, an acid-binding agent is added to the above reaction system, and the reaction continues at 20–30°C for 10–15 h in the presence of a protective gas and in the dark. The product after the reaction is completed is dialyzed and lyophilized to obtain the PEG polyphenol compound.

[0100] S2. Preparation of metal polyphenol nanoparticles containing anti-breast cancer drugs

[0101] The metal salt solution and the PEG polyphenol compound solution were mixed and stirred at 20–30°C for 0.5–1 h; then, the proanthocyanidin solution was added to the above system and stirred at 20–30°C for 5–10 min; then, the gefitinib aqueous solution was added to the above system and stirred at 20–30°C for 4–8 h; after the reaction was completed, the mixture was centrifuged and washed to obtain the metal polyphenol nanoparticles containing the anti-breast cancer drug.

[0102] Fifthly, the present invention provides the use of metal polyphenol nanoparticles as described in the first aspect, or metal polyphenol nanoparticles containing anti-breast cancer drugs as described in the third aspect, in the preparation of TMEM16A and / or EGFR inhibitors.

[0103] TMEM16A (ANO1) is a calcium-activated chloride channel with ten transmembrane segments. TMEM16A ion channels are highly expressed in breast cancer tissues, and this high expression is closely related to cancer proliferation, migration, and invasion. Studies have shown that TMEM16A activation promotes the in vitro migration and invasion of breast cancer cells, as well as the metastasis of breast cancer in mice. Furthermore, breast cancer patients with higher TMEM16A levels have larger lymph node metastases and shorter survival times. Mechanistically, TMEM16A promotes migration and invasion by activating EGFR / STAT3 / ROCK1 signaling. Simultaneously, activated EGFR also further promotes TMEM16A expression.

[0104] Based on this, this invention uses a TMEM16A inhibitor in combination with an EGFR inhibitor to synergistically inhibit breast cancer proliferation, migration, and recurrence through dual targets, thereby achieving highly efficient and specific treatment for breast cancer. In contrast, current broad-spectrum chemotherapy drugs have significant side effects and are prone to drug resistance.

[0105] Sixthly, the present invention provides a method of using nanoparticles as described in the first or third aspect, the method specifically comprising the following steps:

[0106] Metal polyphenol nanoparticles as described in the first aspect, or metal polyphenol nanoparticles containing anti-breast cancer drugs as described in the third aspect, were incubated in the culture medium of MDA-MB-231 cells (human breast cancer cells).

[0107] After the nanoparticles are endocytosed by cells, they are irradiated with a near-infrared laser to induce a photothermal conversion effect and drug release.

[0108] After the medication has taken effect, assess the treatment effectiveness.

[0109] Compared with the prior art, the present invention has the following beneficial effects:

[0110] (1) In this invention, proanthocyanidins, a small molecule inhibitor of natural polyphenols, are used as the backbone and metal ions are coordinated to form a metal polyphenol nanonetwork. Compared with traditional nanoparticles, metal polyphenol nanoparticles with proanthocyanidins as the backbone do not require the introduction of additional carriers and can form a nanonetwork after coordination with metals.

[0111] (2) The metal polyphenol nanoparticles described in this invention have good photothermal responsiveness and can rapidly heat up under near-infrared light irradiation inside or outside the cell. They can also efficiently encapsulate anti-breast cancer drugs, such as the EGFR inhibitor gefitinib. The photothermal properties of the metal polyphenol nanonetwork are used to achieve chemotherapy-PTT synergistic therapy.

[0112] (3) The raw materials of the nanoparticles of the present invention are all non-toxic and highly biocompatible. For example, proanthocyanidins are compounds with definite pharmacological effects found in natural plants. Proanthocyanidins have antioxidant and free radical scavenging capabilities and no toxic side effects. PEG can significantly increase the blood circulation time of nanoparticles and their accumulation at tumor sites. Iron ions are essential metal elements for the human body and are easily metabolized.

[0113] (4) The preparation process of this invention is simple, pollution-free, low-cost, highly efficient, and easy to achieve mass production, and has broad application prospects. Attached Figure Description

[0114] To more clearly illustrate the technical solutions in the specific embodiments of the present invention or the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0115] Figure 1 This is a schematic diagram illustrating the preparation process of the metal polyphenol nanoparticles containing anti-breast cancer drugs described in this invention.

[0116] Figure 2 Scanning electron microscope images of the nanoparticles provided in Examples 1 and 2.

[0117] Figure 3 Measurement graphs of the particle size and potential of the nanoparticles provided in Examples 1 and 2.

[0118] Figure 4 Infrared thermal images of PFG-NPs nanoparticles at different concentrations.

[0119] Figure 5 Infrared thermal images of different nanoparticles.

[0120] Figure 6 The survival rate of MDA-MB-231 cells after different treatment groups is shown in the figure.

[0121] Figure 7 Figures showing the results of live and dead staining experiments with U87mg after different treatment groups. Detailed Implementation

[0122] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0123] Unless otherwise stated, the technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. Furthermore, any methods or materials similar to or equivalent to those described herein may also be used in this invention.

[0124] To facilitate a clearer understanding of the present invention, specific embodiments are described in detail below. Unless otherwise specified, the experimental animals, drugs, and reagents used in the embodiments of the present invention are all sourced from legitimate and readily available channels.

[0125] Example 1

[0126] This embodiment provides proanthocyanidin-Fe nanoparticles (PF-NPs) and proanthocyanidin-Fe-gefitinib nanoparticles (PFG-NPs), which are prepared by the following steps:

[0127] Synthesis of S1 and PEG polyphenol compounds

[0128] 100 mg of 8ARM-PEG-NHS (Pengshuo Biotechnology) and 38 mg of dopamine hydrochloride were added to 25 mL of N,N-dimethylformamide (DMF) and reacted at room temperature for 1 h under nitrogen protection. Then, 35 μL of triethylamine (TEA, Et3N) was added to the above reaction and the reaction was continued for 12 h under light-protected and nitrogen-protected conditions. The product after the reaction was completed was dialyzed under slightly acidic conditions (2 mL of pure water + 20 mL of 1 mol / L hydrochloric acid) for 48 h and then lyophilized at -70 °C for 48 h to obtain PEG polyphenol compound.

[0129] Synthesis of S2, Proanthocyanidin-Fe Nanoparticles (PF-NPs)

[0130] 0.5 mL (100 mg / mL) of ferric chloride hexahydrate (FeCl3·6H2O) aqueous solution and 22 mL (2.6 mg / mL) of PEG polyphenol derivative aqueous solution were stirred at room temperature for 1 h; then 2.5 mL (10 mg / mL) of proanthocyanidin aqueous solution was added, and stirring was continued at room temperature in the dark for 7 h; after the reaction was completed, the PF-NPs nanoparticles were centrifuged (10000 rpm) for 10 min and washed three times with water; finally, the product was redispersed in water at a mass ratio of 1:1 and sealed and stored in a refrigerator at 4 °C in the dark.

[0131] Synthesis of S2', Proanthocyanidin-Fe-Gefitinib Nanoparticles (PFG-NPs)

[0132] 0.5 mL (100 mg / mL) of ferric chloride hexahydrate (FeCl3·6H2O) aqueous solution and 22 mL (2.6 mg / mL) of PEG polyphenol derivative aqueous solution were stirred at room temperature for 1 h. Then, 2.5 mL (10 mg / mL) of proanthocyanidin aqueous solution was added, and the mixture was stirred at room temperature in the dark for 7 h. Then, 1.25 mL (10 mg / mL) of gefitinib aqueous solution was added to the above system, and the mixture was stirred at room temperature in the dark for 7 h. After the reaction was completed, the PFG-NPs nanoparticles were centrifuged (10,000 rpm) for 10 min and washed three times with water. Finally, the product was redispersed in water at a mass ratio of 1:1 and stored in a sealed container at 4 °C in the dark.

[0133] Example 2

[0134] This embodiment provides hyaluronic acid-coated proanthocyanidin-Fe nanoparticles (HA-PF-NPs) and hyaluronic acid-coated proanthocyanidin-Fe-gefitinib nanoparticles (HA-PFG-NPs), wherein the nanoparticles are prepared by the following steps:

[0135] Synthesis of S1 and PEG polyphenol compounds

[0136] 100 mg of 8ARM-PEG-NHS (Pengshuo Biotechnology) and 38 mg of dopamine hydrochloride were added to 25 mL of N,N-dimethylformamide (DMF) and reacted at room temperature for 1 h under nitrogen protection. Then, 35 μL of triethylamine (TEA, Et3N) was added to the above reaction and the reaction was continued for 12 h under light-protected and nitrogen-protected conditions. The product after the reaction was completed was dialyzed under slightly acidic conditions (2 mL of pure water + 20 mL of 1 mol / L hydrochloric acid) for 48 h and then lyophilized at -70 °C for 48 h to obtain PEG polyphenol compound.

[0137] Synthesis of S2, Proanthocyanidin-Fe Nanoparticles (PF-NPs)

[0138] 0.5 mL (100 mg / mL) of ferric chloride hexahydrate (FeCl3·6H2O) aqueous solution and 22 mL (2.6 mg / mL) of PEG polyphenol derivative aqueous solution were stirred at room temperature for 1 h; then 2.5 mL (10 mg / mL) of proanthocyanidin aqueous solution was added, and stirring was continued at room temperature in the dark for 7 h; after the reaction was completed, the PF-NPs nanoparticles were centrifuged (10000 rpm) for 10 min and washed three times with water; finally, the product was redispersed in water at a mass ratio of 1:1 and sealed and stored in a refrigerator at 4 °C in the dark.

[0139] S3. Synthesis of hyaluronic acid-coated proanthocyanidin-Fe nanoparticles (HA-PF-NPs)

[0140] 10 mL of PF-NPs nanoparticles were placed in 10 mL of hyaluronic acid solution (1 mg / mL, average molecular weight 10000 Da), stirred at room temperature for 7 h to complete the coating, and then centrifuged to remove impurities to obtain HA-PF-NPs nanoparticles.

[0141] Synthesis of S2', Proanthocyanidin-Fe-Gefitinib Nanoparticles (PFG-NPs)

[0142] 0.5 mL (100 mg / mL) of ferric chloride hexahydrate (FeCl3·6H2O) aqueous solution and 22 mL (2.6 mg / mL) of PEG polyphenol derivative aqueous solution were stirred at room temperature for 1 h. Then, 2.5 mL (10 mg / mL) of proanthocyanidin aqueous solution was added, and the mixture was stirred at room temperature in the dark for another 0.1 h. Then, 1.25 mL (10 mg / mL) of gefitinib aqueous solution was added to the above system, and the mixture was stirred at room temperature in the dark for another 7 h. After the reaction was completed, the PFG-NPs nanoparticles were centrifuged (10,000 rpm) for 10 min and then washed three times with water. Finally, the product was redispersed in water at a mass ratio of 1:1 and stored in a sealed container at 4 °C in the dark.

[0143] Synthesis of S3', Hyaluronic Acid-Coated Proanthocyanidin-Fe Nanoparticles (HA-PF-NPs)

[0144] 10 mL of PFG-NPs nanoparticles were placed in 10 mL of hyaluronic acid solution (1 mg / mL, molecular weight 10000 Da), and stirred at room temperature for 7 h to complete the coating. Then, the nanoparticles were centrifuged to remove impurities and obtain HA-PFG-NPs nanoparticles.

[0145] Test Example 1

[0146] Characterization data testing of nanoparticles

[0147] Test samples: Nanoparticles provided in Examples 1 and 2;

[0148] Test method: Particle size and potential were measured by scanning electron microscopy (SEM).

[0149] like Figure 2 As shown, the particle size of PF-NPs is 50–120 nm; the particle size of PFG-NPs is 70–130 nm; the particle size of HA-PF-NPs is 100–180 nm; and the particle size of HA-PFG-NPs is 100–220 nm.

[0150] like Figure 3 As shown, the metal polyphenol nanoparticles include an organic framework formed by the reaction of proanthocyanidins and PEG polyphenol compounds, and metal nanoparticles modified by metal coordination in the organic framework; and when the PEG polyphenol compound is obtained by the reaction of an eight-arm polyethylene glycol activated ester and dopamine salt; and when the metal nanoparticles are Fe, nanoparticles with more uniform size and better dispersibility can be obtained.

[0151] Test Example 2

[0152] Photothermal Response Test

[0153] Test samples: PF-NPs, PFG-NPs, HA-PF-NPs, and HA-PFG-NPs nanoparticles provided in Examples 1 and 2;

[0154] Test method: Nanoparticles of different concentrations were placed in a 96-well plate and irradiated with 1W light for 5 minutes (808nm, 1W / cm²). 2 Every 60 seconds, an infrared camera was used to record the images. Additionally, a fixed concentration of pure water, proanthocyanidin aqueous solution (PC), gefitinib aqueous solution (Gf), and two types of nanoparticles, PF-NPs and PFG-NPs, were irradiated with light at a power of 1W for 5 minutes (808nm, 1W / cm²). 2 Infrared cameras were used to capture and record images.

[0155] like Figures 4-5 As shown, the nanoparticles of this invention have good photothermal responsiveness and can rapidly heat up inside or outside cells under near-infrared light irradiation, while efficiently delivering gefitinib, thereby achieving chemotherapy-PTT synergistic therapy.

[0156] Test Example 3

[0157] Cell proliferation test

[0158] Test samples: PF-NPs, PFG-NPs, HA-PF-NPs, and HA-PFG-NPs nanoparticles provided in Examples 1 and 2;

[0159] Test method: The effect of [the assay] on the cell proliferation of MDA-MB-231 cells was determined by the CCK-8 assay. MDA-MB-231 cells were cultured in 96-well plates for 24 h.

[0160] The subjects were divided into a normal group and a light-illuminated group (LASER), and then two types of samples (PF-NPs and PFG-NPs) were added at concentrations of 6.25 μg / mL, 12.5 μg / mL, 25 μg / mL, 50 μg / mL, 100 μg / mL and 200 μg / mL, respectively.

[0161] In the light-illuminated group, after 4 hours of incubation with nanomedicine, each well of the cells was irradiated with an 808nm near-infrared laser at a power of 1W for 5 minutes (808nm, 1W / cm²). 2 After the light exposure ends, continue culturing for 48 hours. After culturing, add 10 μL of LCK-8 assay reagent to each well and incubate at 37℃ for 2-4 hours. Then, set the measurement wavelength to 450 nm on a microplate reader and place the 96-well plate inside to measure the absorbance. Relative cell viability is determined by comparison with a control.

[0162] The results are as follows Figure 6 As shown, both PF-NPs and PFG-NPs inhibited the proliferation of MDA-MB-231 cells in a concentration-dependent manner. Among them, PFG-NPs loaded with gefitinib were more effective than PF-NPs containing only proanthocyanidins. Furthermore, the inhibitory effect of both materials was more significant after laser irradiation than that of the control group.

[0163] The results are as follows Figure 7 The results of live and dead cell staining showed that both PF-NPs and PFG-NPs had better killing effects than the groups treated with small molecule proanthocyanidins and gefitinib alone. Among them, PFG-NPs loaded with gefitinib were more effective than PF-NPs containing only proanthocyanidins. Furthermore, the killing effects of both materials were more significant than those of the control group after laser irradiation.

[0164] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A metal polyphenol nanoparticle, characterized in that, The metal polyphenol nanoparticles include an organic framework body formed by the reaction of proanthocyanidins and PEG polyphenol compounds, and metal nanoparticles modified on the organic framework body by metal coordination. The PEG polyphenol compound is obtained by reacting an octagonal polyethylene glycol activated ester with a dopamine salt, wherein the octagonal polyethylene glycol activated ester is an octagonal polyethylene glycol succinimide ester, and the dopamine salt is dopamine hydrochloride; the metal nanoparticles are selected from any one or a combination of at least two of Cu, Zr or Fe. The preparation method of the metal polyphenol nanoparticles includes the following steps: First, an eight-arm polyethylene glycol activated ester, dopamine salt, and solvent are mixed and reacted; then, an acid-binding agent is added to the above reaction system and the reaction is carried out; after the reaction is completed, the PEG polyphenol compound is collected; the mass ratio of the eight-arm polyethylene glycol activated ester to dopamine salt is 10:(3~5); The metal salt solution and the PEG polyphenol compound solution were mixed and stirred for the first time; then mixed with the proanthocyanidin solution and stirred for the second time; after the reaction was completed, the metal polyphenol nanoparticles were collected; the mass ratio of the metal salt, PEG polyphenol compound and proanthocyanidin was (4~6):(5~7):(2~3); the concentration of the metal salt solution was 50~150 mg / mL, the concentration of the PEG polyphenol compound solution was 1~5 mg / mL, and the concentration of the proanthocyanidin solution was 5~15 mg / mL.

2. The metal polyphenol nanoparticles according to claim 1, characterized in that, The molecular weight of the eight-arm polyethylene glycol activated ester is 8000~12000 Da.

3. The metal polyphenol nanoparticles according to claim 1, characterized in that, The metal nanoparticles are Fe.

4. The metal polyphenol nanoparticles according to claim 1, characterized in that, The metal polyphenol nanoparticles also include a hyaluronic acid shell.

5. The metal polyphenol nanoparticles according to claim 4, characterized in that, The molecular weight of the hyaluronic acid is 8000~12000 Da.

6. A method for preparing metal polyphenol nanoparticles according to any one of claims 1 to 5, characterized in that, The preparation method includes the following steps: First, an eight-arm polyethylene glycol activated ester, dopamine salt, and solvent are mixed and reacted; then, an acid-binding agent is added to the above reaction system and the reaction is carried out; after the reaction is completed, the PEG polyphenol compound is collected; the mass ratio of the eight-arm polyethylene glycol activated ester to dopamine salt is 10:(3~5); The metal salt solution and the PEG polyphenol compound solution were mixed and stirred for the first time; then mixed with the proanthocyanidin solution and stirred for the second time; after the reaction was completed, the metal polyphenol nanoparticles were collected; the mass ratio of the metal salt, PEG polyphenol compound and proanthocyanidin was (4~6):(5~7):(2~3); the concentration of the metal salt solution was 50~150 mg / mL, the concentration of the PEG polyphenol compound solution was 1~5 mg / mL, and the concentration of the proanthocyanidin solution was 5~15 mg / mL.

7. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, In the preparation of the PEG polyphenol compound, the solvent is N,N -Dimethylformamide.

8. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, In the preparation of the PEG polyphenol compound, the acid-binding agent is triethylamine.

9. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, The amount of acid-binding agent added is 0.1-1% of the total mass of the eight-arm polyethylene glycol activated ester and dopamine salt.

10. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, Before adding the acid-binding agent to the above reaction system, react at 20~30℃ for 0.5~2 h.

11. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, After adding an acid-binding agent to the above reaction system, react at 20~30℃ for 10~15 h.

12. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, After the reaction is complete, the collected product needs to be dialyzed under slightly acidic conditions.

13. The method for preparing metal polyphenol nanoparticles according to claim 12, characterized in that, The slightly acidic condition is an aqueous solution of hydrochloric acid.

14. The method for preparing metal polyphenol nanoparticles according to claim 13, characterized in that, The concentration of the aqueous hydrochloric acid solution is 0.5~2 mol / L.

15. The method for preparing metal polyphenol nanoparticles according to claim 12, characterized in that, The dialysis time is 42-54 hours.

16. The method for preparing metal polyphenol nanoparticles according to claim 12, characterized in that, The dialysis process also includes lyophilization, wherein the lyophilization temperature is -80 to -40°C and the lyophilization time is 24 to 48 hours.

17. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, The metal polyphenol nanoparticles are PF-NPs nanoparticles.

18. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, The temperature of the first stirring is 20~30℃, and the time is 0.5~2 h.

19. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, The second stirring is performed at a temperature of 20-30°C for 6-8 hours.

20. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, The method for collecting the metal polyphenol nanoparticles is centrifugation, with a centrifugation speed of 8000~12000 r / min and a centrifugation time of 5~15 min.

21. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, The centrifugation process also includes washing, which involves washing with water at least once.

22. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, The product obtained from the washing process needs to be redispersed in water for storage, and the mass ratio of the product to water is 1:(3~5).

23. The method for preparing metal polyphenol nanoparticles according to claim 6, characterized in that, The preparation method of the metal polyphenol nanoparticles also includes coating with a hyaluronic acid shell, specifically including the following steps: PF-NPs nanoparticles were placed in a hyaluronic acid solution, coated by stirring, and then centrifuged to obtain HA-PF-NPs nanoparticles.

24. The method for preparing metal polyphenol nanoparticles according to claim 23, characterized in that, The mass ratio of the PF-NPs nanoparticles to the hyaluronic acid solution is 1:(0.5~1).

25. The method for preparing metal polyphenol nanoparticles according to claim 23, characterized in that, The concentration of the hyaluronic acid solution is 0.5~1 mg / mL.

26. The method for preparing metal polyphenol nanoparticles according to claim 23, characterized in that, The stirring is carried out under light-protected conditions.

27. The method for preparing metal polyphenol nanoparticles according to claim 23 or 26, characterized in that, The stirring temperature is 20~30℃, and the stirring time is 6~8 h.

28. A metal polyphenol nanoparticle containing an anti-breast cancer drug, characterized in that, The metal polyphenol nanoparticles containing anti-breast cancer drugs include: metal polyphenol nanoparticles as described in any one of claims 1 to 5, and anti-breast cancer drugs coated in the metal polyphenol nanoparticle network.

29. The metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 28, characterized in that, The anti-breast cancer drug is an EGFR inhibitor.

30. The metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 29, characterized in that, The EGFR inhibitor is gefitinib.

31. The metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 29, characterized in that, The loading amount of the EGFR inhibitor is 3~8 wt%.

32. The metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 28, characterized in that, The metal polyphenol nanoparticles containing anti-breast cancer drugs also include a hyaluronic acid shell.

33. The metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 32, characterized in that, The molecular weight of the hyaluronic acid is 8000~12000 Da.

34. A method for preparing metal polyphenol nanoparticles containing an anti-breast cancer drug according to any one of claims 28-33, characterized in that, The preparation method includes the following steps: The metal salt solution and the PEG polyphenol compound solution were mixed and stirred for the first time; then mixed with the proanthocyanidin solution and stirred for the second time; then mixed with the gefitinib solution and stirred for the third time; after the reaction was completed, the metal polyphenol nanoparticles containing the anti-breast cancer drug were collected.

35. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 34, characterized in that, The metal polyphenol nanoparticles containing anti-breast cancer drugs are PFG-NPs nanoparticles.

36. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 34, characterized in that, The concentration of the gefitinib solution is 5-15 mg / mL.

37. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 34, characterized in that, The amount of gefitinib solution added accounts for 2-6% of the total volume of the mixed solution.

38. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 34, characterized in that, The temperature of the first stirring is 20~30℃, and the time is 0.5~1 h.

39. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 34, characterized in that, The second stirring is performed at a temperature of 20-30°C for 5-10 minutes.

40. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 34, characterized in that, The third stirring is performed at a temperature of 20-30°C for 4-8 hours.

41. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 34, characterized in that, The method for collecting the metal polyphenol nanoparticles containing anti-breast cancer drugs is centrifugation, with a centrifugation speed of 8000~12000 r / min and a centrifugation time of 10~15 min.

42. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 41, characterized in that, The centrifugation process also includes washing, which involves washing with water at least once.

43. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 42, characterized in that, The product obtained from the washing process needs to be redispersed in water for storage, and the mass ratio of the product to water is 1:(0.8~1.2).

44. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 34, characterized in that, The preparation method of the metal polyphenol nanoparticles containing anti-breast cancer drugs also includes coating with a hyaluronic acid shell, specifically including the following steps: PFG-NPs nanoparticles were placed in a hyaluronic acid solution, coated by stirring, and then centrifuged to obtain HA-PFG-NPs nanoparticles.

45. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 44, characterized in that, The mass ratio of the PFG-NPs nanoparticles to the hyaluronic acid solution is 1:(0.5~1).

46. ​​The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 44, characterized in that, The concentration of the hyaluronic acid solution is 0.5~1 mg / mL.

47. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 44, characterized in that, The stirring is carried out under light-protected conditions.

48. The method for preparing metal polyphenol nanoparticles containing anti-breast cancer drugs according to claim 44 or 47, characterized in that, The stirring temperature is 20~30℃, and the stirring time is 6~8 h.

49. The use of a metal polyphenol nanoparticle according to any one of claims 1 to 5, or a metal polyphenol nanoparticle containing an anti-breast cancer drug according to any one of claims 28 to 33, in the preparation of TMEM16A and / or EGFR inhibitors.

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