CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, nanomedicines for targeted delivery of CuET, and their preparation and application

Abstract

The present invention belongs to the technical field of nanomaterials, and more specifically, relates to CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, nanomedicines for targeted delivery of CuET, and their preparation and application. Hydroxyethyl starch and polydopamine are used to stabilize CuET nanocrystals to form nanoparticles, and a layer of thiolated hydroxyethyl starch coupled with folic acid is also wrapped on the outer layer of the nanoparticles to provide targeting and stabilizing effects. The nanomedicines for targeted delivery of CuET prepared by the present invention can improve the water solubility of CuET, prolong the circulation time of CuET in the blood, increase the enrichment of CuET in the tumor site, enhance the inhibitory effect of CuET on tumor cells and tumor stem cells, and enhance the anti-tumor effect of CuET in vivo. In addition, combined with hyperbaric oxygen therapy, the anti-tumor effect of the nanomedicines for targeted delivery of CuET can be further improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of nanomaterials, and more specifically, relates to CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, nanomedicines for targeted delivery of CuET, and preparation and application thereof. Background Art

[0002] CuET is a coordination compound formed by the reaction of disulfiram, a widely used alcohol abstinence drug in clinical practice, and Cu(Ⅱ) ions. Studies have shown that CuET is the main active ingredient of disulfiram in its anti-cancer function. Disulfiram is first metabolized into DDTC salt after entering the body, and then reacts with Cu(Ⅱ) ions to generate CuET to kill cancer cells. There have been many clinical studies combining disulfiram and Cu(Ⅱ) ions for cancer treatment. However, clinical research results show that its anti-cancer effect is not good. The main reason is that disulfiram is not water soluble, and it is easily metabolized after entering the body, and it cannot effectively form CuET with the function of killing cancer cells at the tumor site. The method of directly using CuET also faces many challenges. CuET is also very poorly soluble in water and is easily cleared in the body. These factors have greatly limited the application of disulfiram and CuET in anti-cancer.

[0003] At present, there are many technologies that use various nanocarriers to load and deliver disulfiram or CuET for tumor treatment. Pu Yuji et al. used a variety of proteins to prepare CuET nanoparticles, which improved the water solubility of CuET. Tang Hanxiao et al. used PVP, PLLA-PEG and other polymers to prepare microspheres encapsulating CuET, which improved the water solubility and stability of CuET. However, these methods all have the following problems: 1. The prepared drug carrier lacks tumor targeting and is difficult to enrich at the tumor site; 2. The preparation process is complicated, the drug loading efficiency is not high, and the drug loading amount is not high; 3. Toxic organic reagents are required in the preparation process, which brings difficulties to impurity removal, purification and quality control. Summary of the invention

[0004] In view of the defects of the prior art, the purpose of the present invention is to provide a CuET nanoparticle co-stabilized by hydroxyalkyl starch and polydopamine, a nanodrug for targeted delivery of CuET, and its preparation and application, so as to solve the technical problems of the prior art that CuET has poor solubility and stability in water, which is not conducive to tumor treatment application.

[0005] To achieve the above objectives, the present invention provides a CuET nanoparticle co-stabilized by hydroxyalkyl starch and polydopamine, which is a nanoparticle formed by hydroxyalkyl starch and polydopamine co-stabilized CuET nanocrystals, wherein hydroxyalkyl starch and polydopamine are coated on the surface of the CuET nanocrystals to form the CuET nanoparticle co-stabilized by hydroxyalkyl starch and polydopamine.

[0006] Preferably, the hydroxyalkyl starch is hydroxymethyl starch, hydroxyethyl starch, hydroxypropyl starch or hydroxybutyl starch, and more preferably hydroxyethyl starch.

[0007] Preferably, the mass ratio of the hydroxyalkyl starch to the CuET nanocrystals is 100:1-100, and the mass ratio of the polydopamine to the CuET nanocrystals is 10:1-10.

[0008] Preferably, the diameter of the nanoparticles is 40 to 300 nanometers.

[0009] According to another aspect of the present invention, a hydroxyalkyl starch and polydopamine co-stabilized CuET nanoparticle dispersion is provided, which comprises the hydroxyalkyl starch and polydopamine co-stabilized CuET nanoparticles, wherein the nanoparticles are nanoparticles formed by hydroxyalkyl starch and polydopamine co-stabilized CuET nanocrystals, wherein hydroxyalkyl starch and polydopamine are coated on the surface of the CuET nanocrystals so that the CuET nanocrystals are stably dispersed in water to form the hydroxyalkyl starch and polydopamine co-stabilized CuET nanoparticle dispersion.

[0010] Preferably, the concentration of the CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine in the dispersion is 0.1-100 mg / mL.

[0011] According to another aspect of the present invention, there is provided a method for preparing the nanoparticle dispersion, comprising the following steps:

[0012] (1) mixing a Cu(II) ion salt solution and a DDTC salt solution in a hydroxyalkyl starch aqueous solution to react with each other to obtain a hydroxyalkyl starch stabilized CuET nanoparticle dispersion;

[0013] (2) mixing the CuET nanoparticle dispersion stabilized by hydroxyalkyl starch in step (1) with dopamine under alkaline conditions to cause polymerization of dopamine, and obtaining the CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch and polydopamine after purification.

[0014] Preferably, in step (1), the concentration of the hydroxyalkyl starch aqueous solution is 0.1 to 100 mg / mL; the concentration of copper (II) ions in the Cu (II) ion salt solution is 0.000177 to 17.7 mg / mL;

[0015] The Cu(II) ion salt is copper nitrate, copper sulfate or copper chloride; the DDTC salt solution is a DDTC sodium salt aqueous solution or a DDTC potassium salt aqueous solution;

[0016] The Cu(II) ion salt solution and the DDTC salt solution are added dropwise to the hydroxyalkyl starch aqueous solution, and stirring is performed while the addition is performed to obtain a CuET nanoparticle dispersion stabilized by the hydroxyalkyl starch.

[0017] Preferably, in step (2), the molar ratio of dopamine to the Cu(II) ions in step (1) is 10:0.427-4.27; the pH range of the alkaline condition is 9-11; and the purification in step (2) is ultrafiltration or dialysis.

[0018] According to another aspect of the present invention, a CuET nanomedicine co-stabilized by hydroxyalkyl starch and modified polydopamine is provided, which is a nanomedicine obtained by modifying the surface of thiolated hydroxyalkyl starch with polydopamine in the CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch.

[0019] Preferably, the preparation method of the nano drug comprises the following steps: mixing the CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch and polydopamine with thiolated hydroxyalkyl starch and reacting the mixture so that the surface of the polydopamine therein is modified with the thiolated hydroxyalkyl starch, thereby obtaining the nano drug.

[0020] According to another aspect of the present invention, a nano drug for targeted delivery of CuET is provided, which is a nano drug obtained by surface-modifying hydroxyalkyl starch with polydopamine coupled to thiol-modified tumor-targeting molecules in the CuET nanoparticle dispersion co-stabilized with hydroxyalkyl starch and polydopamine.

[0021] Preferably, the preparation method of the nano drug comprises the following steps: mixing the CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch and polydopamine with the thiolated hydroxyalkyl starch coupled with the tumor-targeting molecule, and reacting the mixture so that the surface of the polydopamine is modified by the thiolated hydroxyalkyl starch coupled with the tumor-targeting molecule, thereby obtaining the nano drug.

[0022] Preferably, the mass ratio of the thiolated hydroxyalkyl starch coupled with the tumor-targeting molecule to the Cu(II) ions in the CuET is 100:0.00177-17.7.

[0023] According to another aspect of the present invention, there is provided a use of the CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, the CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch and polydopamine, the CuET nanodrug co-stabilized by hydroxyalkyl starch and modified polydopamine and / or the nanodrug for targeted delivery of CuET in the preparation of drugs for treating and / or preventing tumors.

[0024] Preferably, the CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, the CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch and polydopamine, the CuET nanodrug co-stabilized by hydroxyalkyl starch and modified polydopamine and / or the nanodrug for targeted delivery of CuET are used in a hyperbaric oxygen environment.

[0025] In general, the above technical solution conceived by the present invention has the following advantages compared with the prior art:

[0026] Beneficial effects:

[0027] (1) The present invention provides a hydroxyalkyl starch and polydopamine co-stabilized CuET nanoparticle and nanoparticle dispersion. Hydroxyalkyl starch and polydopamine are used to coat the surface of CuET nanocrystals to form hydroxyalkyl starch and polydopamine co-modified CuET nanoparticles. Experiments have found that this modification method solves the water solubility problem of CuET and improves the solubility and stability of CuET in aqueous solution.

[0028] (2) Based on the CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, the present invention modifies the surface of polydopamine with a layer of thiol-modified hydroxyethyl starch. Experimental results show that the obtained CuET nanoparticles have better stability in the blood and better tumor inhibition effect than those without modifying the polydopamine.

[0029] (3) Based on the CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, the present invention further modifies the surface of polydopamine with a layer of thiol-modified hydroxyethyl starch coupled with tumor-targeting molecules, thereby obtaining a CuET nanomedicine capable of targeted delivery. The CuET nanomedicine can be specifically enriched at the tumor site and is more taken up by tumor cells. Compared with nanomedicines that are not coupled with tumor-targeting molecules, the CuET nanomedicine has a better anti-tumor effect.

[0030] (4) The present invention uses hydroxyethyl starch, which is widely used in clinical practice, as an excipient, and prepares CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, nanoparticle dispersions, CuET nanomedicines co-stabilized by hydroxyalkyl starch and modified polydopamine, and nanomedicines for targeted delivery of CuET in an aqueous solution by a simple preparation method. The preparation process does not contain toxic organic or inorganic reagents and has great clinical application prospects.

[0031] (5) The preparation process of the CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, nanoparticle dispersion, CuET nanodrug co-stabilized by hydroxyalkyl starch and modified polydopamine, and nanodrug for targeted delivery of CuET provided by the present invention has high drug loading efficiency, no complicated impurity removal and purification process, and can be used for the application of anticancer drugs. The encapsulation rate is 100%, the drug loading amount is easy to control, and it is easy to repeat.

[0032] (6) The nanomedicine for targeted delivery of CuET designed and prepared in the preferred embodiment of the present invention solves the water solubility problem of CuET, and molecules modified with tumor targeting groups such as folic acid can be specifically enriched at the tumor site and more taken up by tumor cells.

[0033] (7) The co-stabilized CuET nanomedicine designed and prepared by the present invention can not only kill a variety of cancer cells, but also kill tumor stem cells and inhibit the development of cancer.

[0034] (8) Experiments have shown that the co-stabilized CuET nanomedicine provided by the present invention combined with hyperbaric oxygen therapy for cancer treatment has a synergistic effect.

[0035] (9) A nano drug for targeted delivery of CuET provided in a preferred embodiment of the present invention comprises nanoparticles composed of CuET nanocrystals stabilized by hydroxyethyl starch and polydopamine, and a layer of thiolated hydroxyethyl starch coupled with folic acid is also coated on the outer layer of the nanoparticles to provide targeting and stabilizing effects. First, CuET nanoparticles stabilized by hydroxyethyl starch are prepared by a one-step method, then dopamine is loaded into the nanoparticles by polymerization reaction, and finally the thiolated hydroxyethyl starch coupled with folic acid is coated on the outer layer of the nanoparticles to further enhance the stability and targeting of the CuET nanoparticles. A nano drug for targeted delivery of CuET can be prepared through the above process. The nano drug for targeted delivery of CuET prepared by the present invention can improve the water solubility of CuET, prolong the circulation time of CuET in the blood, increase the enrichment of CuET in the tumor site, enhance the inhibitory effect of CuET on tumor cells and tumor stem cells, and enhance the anti-tumor effect of CuET in vivo. In addition, combined with hyperbaric oxygen therapy, the anti-tumor effect of the nano drug for targeted delivery of CuET can be further enhanced. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 The synthetic route of thiolated hydroxyethyl starch for coupling folic acid;

[0037] Figure 2 These are photos of different samples prepared in Example 1 and Comparative Example 1;

[0038] Figure 3 The dynamic light scattering particle size distribution results of samples 1 to 5;

[0039] Figure 4 TEM results of sample 2;

[0040] Figure 5 is the FTEM result of sample 2;

[0041] Figure 6 The particle size distribution results of different samples in 50% FBS solution;

[0042] Figure 7 XRD results of different samples;

[0043] Figure 8 The UV absorption spectrum analysis results of different samples;

[0044] Fig. 9 The results of 4T1 cell viability after incubation with different concentrations of drugs;

[0045] Fig.10 Results of 4T1 stem cell viability after incubation of different samples with different concentrations of drugs;

[0046] Fig.11 The efficacy results of different samples in treating 4T1 breast cancer subcutaneous tumors;

[0047] Fig.12 The efficacy results of different samples in treating Panc02 pancreatic cancer;

[0048] Fig.13 Photos of Panc02 pancreatic cancer tumors treated with different samples combined with hyperbaric oxygen therapy. DETAILED DESCRIPTION

[0049] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0050] Hydroxyethyl starch is widely used as a plasma expander in clinical practice, and has good water solubility and excellent biocompatibility. In order to improve the stability of CuET in water, the present invention initially attempts to use hydroxyethyl starch for stabilization. Experiments have found that hydroxyethyl starch can improve the solubility and stability of CuET in water to a certain extent, and can be used for cancer treatment by in vivo injection. However, when it is used as an intravenous drug, no obvious tumor inhibition effect is observed, indicating that its stability cannot meet the requirements of intravenous injection. To this end, the present invention further provides a hydroxyalkyl starch and polydopamine co-stabilized CuET nanoparticles, which are nanoparticles formed by hydroxyalkyl starch and polydopamine co-stabilizing CuET nanocrystals, wherein hydroxyalkyl starch and polydopamine are coated on the surface of the CuET nanocrystals to form the hydroxyalkyl starch and polydopamine co-stabilized CuET nanoparticles. The present invention is by simultaneously coating hydroxyalkyl starch and polydopamine on the surface of CuET nanocrystals, and experiments have found that the solubility and stability of CuET nanocrystals in aqueous solution are significantly improved, and can be used for cancer treatment by intravenous injection.

[0051] The hydroxyalkyl starch of the present invention is hydroxymethyl starch, hydroxyethyl starch, hydroxypropyl starch or hydroxybutyl starch, preferably hydroxyethyl starch. In some embodiments, the molecular weight of the hydroxyalkyl starch is 25-480 kDa, and the molar substitution degree of the hydroxyalkyl of the hydroxyalkyl starch is 0.4-0.6; for example, when the hydroxyalkyl starch is hydroxyethyl starch, the molecular weight of the hydroxyethyl starch is 25-480 kDa, and the molar substitution degree of the hydroxyethyl of the hydroxyethyl starch is 0.4-0.6.

[0052] In some embodiments, the mass ratio of the hydroxyalkyl starch to the CuET nanocrystals is 100:1-100, preferably 100:5-20, and the mass ratio of the polydopamine to the CuET nanocrystals is 10:1-10, preferably 10:3-5.

[0053] In some embodiments, the diameter of the nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine provided by the present invention is 40 to 300 nanometers.

[0054] The present invention also provides a CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch and polydopamine, which comprises CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, wherein the nanoparticles are nanoparticles formed by hydroxyalkyl starch and polydopamine co-stabilizing CuET nanocrystals, wherein hydroxyalkyl starch and polydopamine are coated on the surface of the CuET nanocrystals so that the CuET nanocrystals are stably dispersed in water to form the dispersion.

[0055] In some embodiments, the concentration of the CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine in the dispersion is 0.1 to 100 mg / mL, preferably 1 to 10 mg / mL.

[0056] In the present invention, hydroxyalkyl starch and polydopamine are used as stabilizers to coat CuET nanocrystals, which may prevent the aggregation and precipitation of CuET nanocrystals by virtue of the weak binding effect between hydroxyalkyl starch, polydopamine and CuET, thereby improving the solubility and stability of CuET nanocrystals in aqueous solution.

[0057] The present invention also provides a method for preparing the nanoparticle dispersion, comprising the following steps:

[0058] (1) mixing a Cu(II) ion salt solution and a DDTC salt solution in a hydroxyalkyl starch aqueous solution to react with each other to obtain a hydroxyalkyl starch stabilized CuET nanoparticle dispersion;

[0059] (2) mixing the CuET nanoparticle dispersion stabilized by hydroxyalkyl starch in step (1) with dopamine under alkaline conditions to cause polymerization of dopamine, and obtaining the CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch and polydopamine after purification.

[0060] In some embodiments, in step (1), the concentration of the hydroxyalkyl starch aqueous solution is 0.1-100 mg / mL; the concentration of copper (II) ions in the Cu (II) ion salt solution is 0.000177-17.7 mg / mL; the molar ratio of the DDTC ions to the Cu (II) ions is 1.8-2.2:1, preferably 2:1; the Cu (II) ion salt is copper nitrate, copper sulfate or copper chloride; the DDTC salt solution is a DDTC sodium salt aqueous solution or a DDTC potassium salt aqueous solution; the Cu (II) ion salt solution and the DDTC salt solution are added dropwise to the hydroxyalkyl starch aqueous solution, and stirred while dropping to obtain a hydroxyalkyl starch stabilized CuET nanoparticle dispersion. The stirring includes but is not limited to magnetic stirring, mechanical stirring, vortex oscillation or ultrasonic oscillation.

[0061] In some embodiments, the Cu(II) ion salt solution and the DDTC salt solution are added to the hydroxyalkyl starch aqueous solution by dropwise addition, and the dropwise addition speed is 0.01 to 10 mL / s. When the stirring method is magnetic stirring, mechanical stirring, or vortex oscillation, the stirring speed is 10 to 3000 rpm / min. When the stirring method is ultrasonic oscillation, the ultrasonic power is 10 to 200 W.

[0062] In some embodiments, in step (2), the molar ratio of dopamine to the Cu(II) ions in step (1) is 10:0.427-4.27; after the hydroxyalkyl starch-stabilized CuET nanoparticle dispersion is mixed with dopamine in step (2), ammonia water or sodium hydroxide is also introduced to obtain the alkaline environment, and the pH range is 9-11. The purification in step (2) is ultrafiltration or dialysis.

[0063] In order to further improve the stability of CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine in the blood, the present invention modifies the surface of polydopamine in the nanoparticles with thiolated hydroxyalkyl starch, so that polydopamine reacts with thiol to form a CS covalent bond, further improving the stability of the nanodrug in the blood, and provides a CuET nanodrug co-stabilized by hydroxyalkyl starch and modified polydopamine. In some embodiments, the dispersion of CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine is mixed with thiolated hydroxyalkyl starch and then reacted, so that the surface of polydopamine therein is modified with thiolated hydroxyalkyl starch, that is, the CuET nanodrug co-stabilized by hydroxyalkyl starch and modified polydopamine is obtained. In some embodiments, the mass ratio of the thiolated hydroxyalkyl starch to the Cu(II) ions in the CuET is 100:0.00177~17.7.

[0064] In some embodiments, the thiolated hydroxyalkyl starch can be prepared by three steps: step S1, reacting hydroxyethyl starch with succinic anhydride to obtain carboxylated hydroxyethyl starch; step S2, reacting carboxylated hydroxyethyl starch with 2-(pyridine disulfide)-ethylamine hydrochloride to obtain hydroxyethyl starch-2-(pyridine disulfide); step S3, thiolation of hydroxyethyl starch-2-(pyridine disulfide) to obtain thiolated hydroxyalkyl starch.

[0065] Furthermore, in order to improve the targeting of CuET nanomedicine, the present invention also provides a nanomedicine for targeted delivery of CuET, which is a nanomedicine obtained by modifying the surface of polydopamine in the CuET nanoparticles co-stabilized with hydroxyalkyl starch and polydopamine with a layer of thiolated hydroxyalkyl starch coupled with a tumor-targeting molecule. The tumor-targeting molecule includes but is not limited to folic acid, lactobionic acid or iRGD peptide.

[0066] Taking folic acid as an example of a tumor-targeting molecule, thiolated hydroxyalkyl starch coupled with folic acid can be prepared by four steps: step S1, coupling hydroxyethyl starch with folic acid through an ester bond; step S2, reacting the hydroxyethyl starch coupled with folic acid with succinic anhydride to obtain carboxylated hydroxyethyl starch coupled with folic acid; step S3, reacting the carboxylated hydroxyethyl starch coupled with folic acid with 2-(pyridine disulfide)-ethylamine hydrochloride to obtain hydroxyethyl starch-2-(pyridine disulfide) coupled with folic acid; step S4, thiolation of hydroxyethyl starch-2-(pyridine disulfide) coupled with folic acid to obtain thiolated hydroxyalkyl starch coupled with folic acid, and the reaction route is as follows. Figure 1 shown.

[0067] In some embodiments, the preparation method of the nanodrug comprises the following steps: mixing the CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch and polydopamine with the thiolated hydroxyalkyl starch coupled with the tumor-targeting molecule and reacting the mixture so that the surface of the polydopamine is modified with a layer of thiolated hydroxyalkyl starch coupled with the tumor-targeting molecule, thereby obtaining the nanodrug.

[0068] In some embodiments, the mass ratio of the thiolated hydroxyalkyl starch coupled with folic acid to the Cu(II) ion in the CuET is 100:0.00177-17.7. The reaction time after mixing is 4-48 hours.

[0069] The CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, the CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch and polydopamine, the CuET nanomedicine co-stabilized by hydroxyalkyl starch and modified polydopamine and / or the nanomedicine for targeted delivery of CuET described in the present invention can be used to prepare drugs for treating and / or preventing tumors.

[0070] Hyperbaric oxygen is widely used in the clinic as a treatment for carbon monoxide poisoning. The present invention has found that hyperbaric oxygen also helps promote the anti-tumor effect of anti-tumor nanomedicines. The present invention combines CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine, CuET nanoparticle dispersions co-stabilized by hydroxyalkyl starch and polydopamine, CuET nanomedicine co-stabilized by hydroxyalkyl starch and modified polydopamine, and / or the nanomedicine for targeted delivery of CuET with hyperbaric oxygen for anti-tumor treatment of mice, achieving good anti-tumor effects.

[0071] The following are examples:

[0072] Example 1

[0073] This embodiment provides a nano drug for targeted delivery of CuET, wherein 200 / 0.5 specifications of hydroxyethyl starch are selected, and CuCl2 and DDTC sodium salt aqueous solutions are used as reaction ion solutions, and the following preparation steps are performed:

[0074] (1) Dissolve 10 mg of hydroxyethyl starch in 1 mL of water, add 1 mL of a 0.374 mg / mL CuCl2 aqueous solution, and mix at 25° C. to obtain a mixed solution A; wherein the molecular weight of the hydroxyethyl starch is 200 kDa, and the degree of hydroxyethyl substitution is 0.5;

[0075] (2) 1 mL of 0.954 mg / mL DDTC sodium salt aqueous solution was added dropwise to the obtained mixed solution A while mixing, wherein the dropping time was 60 s, wherein the mixing method was magnetic stirring at 1000 rpm / min, and the mixing temperature was 25° C. After the dropwise addition was completed, a mixed solution B was obtained. The mixed solution B is a CuET nanoparticle solution stabilized by hydroxyethyl starch.

[0076] (3) Add 1 mL of 2.5 mg / mL dopamine aqueous solution to the obtained mixed solution B, and then add 1 mL of ammonia water and 5 mL of ultrapure water. Stir magnetically at 25°C for 8 h at a stirring speed of 1000 rpm / min to obtain a mixed solution C. Place the mixed solution C in a 3500 Da dialysis bag and dialyze with ultrapure water for 3 days to obtain a mixed solution D. The mixed solution D is a CuET nanoparticle dispersion stabilized by hydroxyethyl starch and polydopamine.

[0077] (4) Add 10 mg of thiolated hydroxyethyl starch coupled with folic acid to the obtained mixed solution D, and stir magnetically for 48 hours at a speed of 1000 rpm / min. The nanomedicine for targeted delivery of CuET is obtained, which is named sample 1. The synthesis method of thiolated hydroxyethyl starch coupled with folic acid, hydroxyethyl starch 200 / 0.5 is selected, and the preparation process is as follows:

[0078] S1: 1 g of hydroxyethyl starch was dissolved in 10 mL of DMSO, and 0.1 g of folic acid, 0.32 g of EDCI and 0.21 g of DMAP were added, and the mixture was reacted at 45°C for 48 h. The reaction product was placed in a dialysis bag with a molecular weight cutoff of 3500 Da, dialyzed in ultrapure water for 3 days, and the sample was freeze-dried at -50°C for 96 h to obtain product 1.

[0079] S2: Dissolve 1 g of product 1 in 10 mL of DMSO, add 0.14 g of succinic anhydride and 0.2 g of DMAP, and react at 45°C for 24 h. Place the reaction product in a dialysis bag with a molecular weight cutoff of 3500 Da, dialyze in ultrapure water for 3 days, and freeze-dry the sample at -50°C for 96 h to obtain product 2.

[0080] S3: Dissolve 1 g of product 2 in 30 mL of ultrapure water, add 0.38 g of EDCI, 0.12 g of NHS and 0.22 g of 2-(2-pyridyldisulfide)ethylamine hydrochloride, react at 25°C for 48 h, place the reaction product in a dialysis bag with a molecular weight cutoff of 3500 Da, dialyze in ultrapure water for 3 days, freeze-dry the sample at -50°C for 96 h to obtain product 3.

[0081] S4: 0.5 g of product 4 was dissolved in 15 mL of DMSO, 0.8 g of dithiothreitol was added, and the mixture was reacted at 25° C. for 24 h under nitrogen protection. The reaction product was placed in a dialysis bag with a molecular weight cutoff of 3500 Da, and dialyzed in ultrapure water for 3 days. The sample was freeze-dried at -50° C. for 96 h to obtain product 4. Product 4 is thiolated hydroxyethyl starch coupled with folic acid.

[0082] Comparative Example 1

[0083] The preparation of CuET includes the following steps:

[0084] 1 mL of 0.954 mg / mL DDTC sodium salt aqueous solution was added dropwise to 1 mL of 0.374 mg / mL CuCl2 aqueous solution, wherein the dropping time was 60 s, wherein the mixing method was magnetic stirring at 1000 rpm / min, and the mixing temperature was 25° C. After the dropping was completed, the CuET sample was obtained.

[0085] Figure 2 From the photographs of the CuET sample prepared in this comparative example, the mixed solution B obtained in step (2) of Example 1, the mixed solution D obtained in step (3) of Example 1, and the sample 1 prepared in Example 1 after being placed for 12 hours, it can be seen that the CuET sample prepared in this comparative example has obvious agglomeration and precipitation of CuET, while the mixed solution B, the mixed solution D, and the sample 1 in Example 1 are all stable dispersions.

[0086] Example 2

[0087] This embodiment provides a nano drug for targeted delivery of CuET, wherein 200 / 0.5 specifications of hydroxyethyl starch are selected, and CuCl2 and DDTC sodium salt aqueous solutions are used as reaction ion solutions, and the following preparation steps are performed:

[0088] (1) Dissolve 10 mg of hydroxyethyl starch in 1 mL of water, add 1 mL of a 0.374 mg / mL CuCl2 aqueous solution, and mix at 25° C. to obtain a mixed solution A; wherein the molecular weight of the hydroxyethyl starch is 200 kDa, and the degree of hydroxyethyl substitution is 0.5;

[0089] (2) Add 1 mL of 0.954 mg / mL DDTC sodium salt aqueous solution dropwise to the obtained mixed solution A while mixing, wherein the dropping time is 60 s, wherein the mixing method is magnetic stirring at 1000 rpm / min, and the mixing temperature is 25° C. After the dropwise addition is completed, a mixed solution B is obtained. The mixed solution B is a CuET nanoparticle solution stabilized by hydroxyethyl starch.

[0090] (3) Add 1 mL of 2.5 mg / mL dopamine aqueous solution to the obtained mixed solution B, and then add 1 mL of ammonia water and 5 mL of ultrapure water. Stir magnetically at 25°C for 8 h at a stirring speed of 1000 rpm / min to obtain a mixed solution C. Place the mixed solution C in a 3500 Da dialysis bag and dialyze with ultrapure water for 3 days to obtain a mixed solution D. The mixed solution D is a CuET nanoparticle dispersion stabilized by hydroxyethyl starch and polydopamine.

[0091] (4) Add 10 mg of thiolated hydroxyethyl starch coupled with folic acid (preparation steps are the same as in Example 1) to the obtained mixed solution D, and stir magnetically for 48 hours at a speed of 1000 rpm / min. Thus, a nanomedicine for targeted delivery of CuET was obtained, which was named Sample 2.

[0092] Example 3

[0093] This embodiment provides a nano drug for targeted delivery of CuET. Hydroxyethyl starch is selected as 130 / 0.4 specification, and CuCl2 and DDTC sodium salt aqueous solution are used as reaction ion solutions, and the following preparation steps are performed:

[0094] (1) Dissolve 10 mg of hydroxyethyl starch in 1 mL of water, add 1 mL of a 0.374 mg / mL CuCl2 aqueous solution, and mix at 60° C. to obtain a mixed solution A; wherein the molecular weight of the hydroxyethyl starch is 130 kDa, and the degree of hydroxyethyl substitution is 0.4;

[0095] (2) Add 1 mL of 0.954 mg / mL DDTC sodium salt aqueous solution dropwise to the obtained mixed solution A while mixing, wherein the dropping time is 60 s, wherein the mixing method is magnetic stirring at 1000 rpm / min, and the mixing temperature is 60° C. After the dropwise addition is completed, a mixed solution B is obtained. The mixed solution B is a CuET nanoparticle solution stabilized by hydroxyethyl starch.

[0096] (3) Add 1 mL of 2.5 mg / mL dopamine aqueous solution to the obtained mixed solution B, and then add 1 mL of ammonia water and 5 mL of ultrapure water. Stir magnetically at 25°C for 8 h at a stirring speed of 1000 rpm / min to obtain a mixed solution C. Place the mixed solution C in a 3500 Da dialysis bag and dialyze with ultrapure water for 3 days to obtain a mixed solution D. The mixed solution D is a CuET nanoparticle dispersion stabilized by hydroxyethyl starch and polydopamine.

[0097] (4) Add 10 mg of thiolated hydroxyethyl starch coupled with folic acid (preparation steps are the same as in Example 1) to the obtained mixed solution D, and stir magnetically for 48 hours at a speed of 1000 rpm / min. Thus, a nano drug for targeted delivery of CuET was obtained, named Sample 3.

[0098] Example 4

[0099] This embodiment provides a nano drug for targeted delivery of CuET. Hydroxyethyl starch is selected as 480 / 0.5 specification, and CuCl2 and DDTC sodium salt aqueous solution are used as reaction ion solutions, and the following preparation steps are performed:

[0100] (1) Dissolve 10 mg of hydroxyethyl starch in 1 mL of water, add 1 mL of a 0.374 mg / mL CuCl2 aqueous solution, and mix at 25° C. to obtain a mixed solution A; wherein the molecular weight of the hydroxyethyl starch is 480 kDa, and the degree of hydroxyethyl substitution is 0.5;

[0101] (2) Add 1 mL of 0.954 mg / mL DDTC sodium salt aqueous solution dropwise to the obtained mixed solution A while mixing, wherein the dropping time is 60 s, wherein the mixing method is magnetic stirring at 1000 rpm / min, and the mixing temperature is 25° C. After the dropwise addition is completed, a mixed solution B is obtained. The mixed solution B is a CuET nanoparticle solution stabilized by hydroxyethyl starch.

[0102] (3) Add 1 mL of 2.5 mg / mL dopamine aqueous solution to the obtained mixed solution B, and then add 1 mL of ammonia water and 5 mL of ultrapure water. Stir magnetically at 25°C for 8 h at a stirring speed of 1000 rpm / min to obtain a mixed solution C. Place the mixed solution C in a 3500 Da dialysis bag and dialyze with ultrapure water for 3 days to obtain a mixed solution D. The mixed solution D is a CuET nanoparticle dispersion stabilized by hydroxyethyl starch and polydopamine.

[0103] (4) Add 10 mg of thiolated hydroxyethyl starch coupled with folic acid (preparation steps are the same as in Example 1) to the obtained mixed solution D, and stir magnetically for 48 hours at a speed of 1000 rpm / min. Thus, a nano drug for targeted delivery of CuET was obtained, named Sample 4.

[0104] Example 5

[0105] This embodiment provides a nano drug for targeted delivery of CuET, wherein 200 / 0.5 specifications of hydroxyethyl starch are selected, and CuCl2 and DDTC sodium salt aqueous solutions are used as reaction ion solutions, and the following preparation steps are performed:

[0106] (1) Dissolve 10 mg of hydroxyethyl starch in 1 mL of water, add 1 mL of a 0.374 mg / mL CuCl2 aqueous solution, and mix at 25° C. to obtain a mixed solution A; wherein the molecular weight of the hydroxyethyl starch is 200 kDa, and the degree of hydroxyethyl substitution is 0.5;

[0107] (2) Add 1 mL of 0.954 mg / mL DDTC sodium salt aqueous solution dropwise to the obtained mixed solution A while mixing, wherein the dropping time is 60 s, wherein the mixing method is magnetic stirring at 1000 rpm / min, and the mixing temperature is 25° C. After the dropwise addition is completed, a mixed solution B is obtained. The mixed solution B is a CuET nanoparticle solution stabilized by hydroxyethyl starch.

[0108] (3) Add 1 mL of 2.5 mg / mL dopamine aqueous solution to the obtained mixed solution B, and then add 1 mL of ammonia water and 5 mL of ultrapure water. Stir magnetically at 25°C for 8 h at a stirring speed of 1000 rpm / min to obtain a mixed solution C. Place the mixed solution C in a 3500 Da dialysis bag and dialyze with ultrapure water for 3 days to obtain a mixed solution D. The mixed solution D is a CuET nanoparticle dispersion stabilized by hydroxyethyl starch and polydopamine.

[0109] (4) Add 10 mg of thiolated hydroxyethyl starch to the obtained mixed solution D, and stir magnetically for 48 h at a speed of 1000 rpm / min. Thus, a CuET nanodrug co-stabilized by hydroxyalkyl starch and modified polydopamine was obtained, which was named sample 5. The preparation steps of the thiolated hydroxyethyl starch are as follows:

[0110] S1: 1 g of hydroxyethyl starch (select 200 / 0.5 of hydroxyethyl starch) was dissolved in 10 mL of DMSO, 0.14 g of succinic anhydride and 0.2 g of DMAP were added, and the mixture was reacted at 45°C for 24 h. The reaction product was placed in a dialysis bag with a molecular weight cutoff of 3500 Da, dialyzed in ultrapure water for 3 days, and the sample was freeze-dried at -50°C for 96 h to obtain product 1.

[0111] S2: Dissolve 1 g of product 1 in 30 mL of ultrapure water, add 0.38 g of EDCI, 0.12 g of NHS and 0.22 g of 2-(2-pyridyldisulfide)ethylamine hydrochloride, react at 25°C for 48 h, place the reaction product in a dialysis bag with a molecular weight cutoff of 3500 Da, dialyze in ultrapure water for 3 days, freeze-dry the sample at -50°C for 96 h to obtain product 2.

[0112] S3: 0.5 g of product 2 was dissolved in 15 mL of DMSO, 0.8 g of dithiothreitol was added, and the mixture was reacted at 25°C for 24 h under nitrogen protection. The reaction product was placed in a dialysis bag with a molecular weight cutoff of 3500 Da, and dialyzed in ultrapure water for 3 days. The sample was freeze-dried at -50°C for 96 h to obtain product 3. Product 3 is thiolated hydroxyethyl starch.

[0113] Experimental Example 1

[0114] Morphological determination of nanomedicine for targeted delivery of CuET: The solutions of samples 1 to 5 prepared in Examples 1 to 5 were taken and the particle size and distribution of the nanoparticles were determined on a dynamic light scattering instrument. Sample 2 prepared in Example 2 was dripped onto the copper mesh carbon film, and after drying, its morphology and element distribution were observed on TEM and FTEM. Figure 3 The results show that the particle size distribution of samples 1 to 5 prepared in Examples 1 to 5 is relatively uniform, and the distribution range is 40 to 200 nanometers. Among them, solution D of Example 1 and Example 5 is a CuET nanoparticle dispersion stabilized by hydroxyethyl starch and polydopamine. After being modified with thiolated hydroxyethyl starch in step (4) of Example 5, the diameter of the nanoparticles increases from 74 nanometers to 107 nanometers in sample 5; after being modified with thiolated hydroxyethyl starch coupled with folic acid in step (4) of Example 1, the diameter of the nanoparticles increases from 74 nanometers to 102 nanometers in sample 2. Figure 4 The TEM results of sample 2 show that the prepared nanomedicine for targeted delivery of CuET is a nanoparticle formed by hydroxyethyl starch and polydopamine encapsulating CuET nanocrystals and modifying the surface of thiolated hydroxyethyl starch coupled with folic acid. Figure 5 The elemental distribution of the nanoparticles by FTEM also confirmed this conclusion.

[0115] Experimental Example 2

[0116] The mixed solution B, mixed solution D and sample 2 solution prepared in Example 2 were mixed with 50% FBS (fetal bovine serum), and the particle size and distribution of the nanoparticles in the mixed solution were measured on a dynamic light scattering instrument on days 0, 1, 2, 3, 4, 5, 6 and 7, respectively. Figure 6The results showed that the nanoparticles in mixed solution D (CuET nanoparticle dispersion stabilized by hydroxyethyl starch and polydopamine) and sample 2 were relatively stable, while the nanoparticles in mixed solution B (CuET nanoparticle solution stabilized by hydroxyethyl starch) adsorbed proteins in FBS, which resulted in larger particle size and aggregation.

[0117] Experimental Example 3

[0118] Spectral characterization of nanoparticles: The sample 2 solution prepared in Example 2 was freeze-dried to obtain a solid nanomedicine for targeted delivery of CuET. The solid sample was subjected to XRD to confirm its structure. Figure 7 The XRD results show that in the XRD results of sample 2, the peaks at 10.15°, 11.79°, 12.35°, 14.44°, 21.3°, and 24.14° are consistent with the results of CuET and the standard card, which indicates that CuET exists in the nanoparticles in the form of nanocrystals. Sample solution 2 was measured by ultraviolet absorption spectroscopy. Figure 8 The UV results showed that sample 2 had an absorption peak at 450nm, which further confirmed the presence of CuET in the nanoparticles. Comparison of the UV results of CuET@HES and CuET@PDA / HES showed that there was a significant absorption enhancement at wavelengths of 300-400nm, indicating that PDA was successfully polymerized into the CuET nanoparticles. Comparison of the UV absorption spectra of CuET@PDA / HES and sample 2 showed that the UV absorption peak at 280nm was significantly enhanced, indicating that the thiolated hydroxyethyl starch coupled with folic acid was successfully modified on the surface of the nanoparticles.

[0119] Experimental Example 4

[0120] Cytotoxicity experiment: Breast cancer 4T1 cells were seeded into 96-well plates at a seeding density of 5000 cells / well, with a culture medium volume of 0.1 mL, and incubated overnight in a cell culture incubator (37°C, 5% CO2). The culture medium was removed, and 0.2 mL of culture medium containing different concentrations of drugs was added and incubated for 24 hours. Cell viability was determined using the CCK8 method. Fig. 9 The results showed that sample 2 prepared in Example 2 had the lowest IC50 value, which was 53.50 ng / mL.

[0121] Experimental Example 5

[0122] Stem cell killing experiment: 4T1 stem cells were seeded into 96-well low-adhesion plates at a seeding density of 5000 cells / well, with a culture medium volume of 0.1 mL, and incubated overnight in a cell culture incubator (37°C, 5% CO2). 0.1 mL of culture medium containing different concentrations of drugs was added and incubated for 24 hours. Cell viability was determined using the CCK8 method. Fig.10 The results showed that sample 2 prepared in Example 2 had the lowest IC50 value, which was 64.08 ng / mL.

[0123] Experimental Example 6

[0124] Animal efficacy experiment: The mouse breast cancer 4T1 cell suspension was subcutaneously inoculated on the right posterior side of the back of female BALB / c mice, 4×10 5 cells / 0.1mL, a mouse breast cancer 4T1 subcutaneous transplant tumor model was established. When the subcutaneous tumor volume was 50-120mm 3 At 14:00, mice were randomly divided into 4 groups, 8 mice in each group, and injected with physiological saline, disulfiram + CuCl2, CuET and sample 2 through the tail vein, respectively. The dose of CuET was 4 mg / kg. The first day of administration was marked as day 1. The above dose was administered 5 times through the tail vein on days 3, 5, 7, 9 and 11. From day 1, the long side a and short side b of the subcutaneous tumor of the mice were measured with a vernier caliper every other day. The tumor volume was calculated as: V = a × b × b / 2. The tumor volume-time curve was drawn.

[0125] Fig.11 CuET@HES is the mixed solution B of Example 2, i.e., the CuET nanoparticle solution stabilized by hydroxyethyl starch; CuET is the CuET sample prepared in Comparative Example 1; the DSF+CuCl2 group is a mixed solution of disulfiram DSF and CuCl2 with a molar concentration of 1:1, both of which are purchased from the market, and the molar concentration is consistent with the concentration of CuET; the solution D group is the CuET nanoparticle dispersion stabilized by hydroxyethyl starch and polydopamine in the mixed solution D of Example 2. From the perspective of the tumor inhibition effect on the 15th day, the CuET@HES group, the CuET group and the DSF+CuCl2 group had poor tumor inhibition effects, and did not show any tumor inhibition effect compared with the normal saline group; the solution D group had a certain improvement in the tumor inhibition effect compared with the normal saline group; and the sample 5 obtained in Example 5, i.e., the CuET nanodrug stabilized by hydroxyethyl starch and modified polydopamine, had shown obvious tumor inhibition effects, indicating that the modification of polydopamine with thiolated hydroxyethyl starch further improved the stability of the CuET nanodrug in the blood; the sample 2 obtained in Example 2 (the targeted delivery CuET nanodrug after further modification of the thiolated hydroxyethyl starch coupled with folic acid) had the best tumor inhibition effect due to the targeting effect of folic acid.

[0126] Experimental Example 7

[0127] Animal efficacy experiment: The pancreas of male C57 mice was inoculated with mouse pancreatic cancer Panc02 cell suspension, 1×10 6 cells / 0.05mL to establish a mouse pancreatic cancer model. Ten days after the mouse model was established, the mice were randomly divided into three groups, each with 8 mice, and injected with normal saline, disulfiram + CuCl2 and sample 5 through the tail vein, respectively. The dose of CuET was 4 mg / kg, and the first day of administration was recorded as day 1. Then, the above dose was administered five times through the tail vein on days 3, 5, 7, 9, and 11. The mice were killed on day 23, and the pancreatic cancer tumors were removed and weighed. Fig.12 The results showed that compared with the control group, sample 5 prepared in Example 5 had the best tumor inhibition effect.

[0128] Experimental Example 8

[0129] Animal efficacy experiment: The pancreas of male C57 mice was inoculated with mouse pancreatic cancer Panc02 cell suspension, 1×10 6 cells / 0.05mL to establish a mouse pancreatic cancer model. Seven days after mouse modeling, the mice were randomly divided into 6 groups, with 8 mice in each group. The hyperbaric oxygen group was treated with hyperbaric oxygen once on the 7th, 8th and 9th day of mouse modeling, and the hyperbaric oxygen condition was 2.5 atmospheres for 90 minutes. On the 10th day of mouse modeling, normal saline, disulfiram + CuCl2 and sample 5 were injected through the tail vein, where the dose of CuET was 4 mg / kg. The hyperbaric oxygen group treated the mice with hyperbaric oxygen immediately after administration. The first day of administration was recorded as the 1st day, and then the above dose was administered 5 times through the tail vein on the 3rd, 5th, 7th, 9th and 11th days. The mice were killed on the 23rd day, the pancreatic cancer tumors were removed, and photos were taken. Fig.13 The results showed that compared with other control groups, sample 5 prepared in Example 5 had a better tumor inhibition effect, and the effect was even better after combined with hyperbaric oxygen therapy.

[0130] It will be easily understood by those skilled in the art that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A nanomedicine for targeted delivery of CuET, characterized in that: It is a nanomedicine obtained by modifying the surface of polydopamine in a CuET nanoparticle dispersion co-stabilized with hydroxyalkyl starch coupled with a thiolated molecule targeting a tumor; The hydroxyalkyl starch and polydopamine co-stabilized CuET nanoparticle dispersion is a dispersion of hydroxyalkyl starch and polydopamine co-stabilized CuET nanoparticles in an aqueous solution, and the alkyl starch and polydopamine co-stabilized CuET nanoparticles are nanoparticles formed by hydroxyalkyl starch and polydopamine co-stabilized CuET nanocrystals, wherein hydroxyalkyl starch and polydopamine are coated on the surface of the CuET nanocrystals to form the hydroxyalkyl starch and polydopamine co-stabilized CuET nanoparticles; The preparation method of the nano drug comprises the following steps: mixing the CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch and polydopamine with the thiolated hydroxyalkyl starch coupled with the tumor-targeting molecule, and reacting the mixture so that the surface of the polydopamine is modified by the thiolated hydroxyalkyl starch coupled with the tumor-targeting molecule, thereby obtaining the nano drug; the tumor-targeting molecule is folic acid; The hydroxyalkyl starch is hydroxyethyl starch; In the CuET nanoparticles co-stabilized by alkyl starch and polydopamine, the mass ratio of the hydroxyalkyl starch to the CuET nanocrystals is 100:1-100, and the mass ratio of the polydopamine to the CuET nanocrystals is 10:1-10; the diameter of the nanoparticles is 40-300 nanometers.

2. The nano drug according to claim 1, characterized in that In the CuET nanoparticle dispersion co-stabilized by hydroxyalkyl starch and polydopamine, the concentration of the CuET nanoparticles co-stabilized by hydroxyalkyl starch and polydopamine in the dispersion is 0.1-100 mg / mL.

3. The nano drug according to claim 1, characterized in that The method for preparing the nanoparticle dispersion comprises the following steps: (1) mixing a Cu(II) ion salt solution and a DDTC salt solution in a hydroxyalkyl starch aqueous solution to react with each other to obtain a hydroxyalkyl starch stabilized CuET nanoparticle dispersion; (2) The CuET nanoparticle dispersion stabilized by hydroxyalkyl starch in step (1) is mixed with dopamine under alkaline conditions to cause polymerization of dopamine, and the CuET nanoparticle dispersion stabilized by hydroxyalkyl starch and polydopamine is obtained after purification.

4. The nano drug according to claim 3, characterized in that In step (1), the concentration of the hydroxyalkyl starch aqueous solution is 0.1-100 mg / mL; the concentration of copper (II) ions in the Cu (II) ion salt solution is 0.000177-17.7 mg / mL; The Cu (II) ion salt is copper nitrate, copper sulfate or copper chloride; the DDTC salt solution is DDTC sodium salt aqueous solution or DDTC potassium salt aqueous solution; Adding the Cu(II) ion salt solution and the DDTC salt solution to the hydroxyalkyl starch aqueous solution by dropwise addition, stirring while dropping, to obtain a CuET nanoparticle dispersion stabilized by hydroxyalkyl starch; In step (2), the molar ratio of the dopamine to the Cu (II) ions in step (1) is 10:0.427-4.

27.

5. Use of the nanomedicine for targeted delivery of CuET according to any one of claims 1 to 4 in the preparation of a drug for treating and / or preventing tumors, wherein the tumor is breast cancer or pancreatic cancer.

6. The use according to claim 5, characterized in that The nanomedicine for targeted delivery of CuET as claimed in any one of claims 1 to 4 is used in a hyperbaric oxygen environment.

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