Application of a Composite Nanozyme Material in a Tumor Dual-Targeted Drug Delivery System

Abstract

The present disclosure provides an application of a composite nanozyme material in a tumor dual-targeted drug delivery system. The composite nanozyme material is used in the preparation of an anti-tumor injection or an implantable medicament. The composite nanozyme material has peroxidase-like activity, catalase-like activity, and glutathione oxidase-like activity. The application method includes: using the composite nanozyme material as a contrast agent to determine the location of the tumor by magnetic resonance; setting a magnetic field at the location of the tumor and injecting an injection or a medicament containing the composite nanozyme material. The present invention first uses a magnetic composite nanozyme material as a MIR contrast agent to determine the location of the tumor by magnetic resonance technology, and further utilizes the biological activity targeting of the composite nanozyme and the physical magnetic targeting of the external magnetic field of the tumor tissue to achieve the specific release of the chemotherapy drug functional molecule at the target site.

Description

Technical Field

[0001] The present disclosure relates to the field of biomedical technologies, and particularly to the application of a composite nanozyme material in a tumor dual-targeted drug delivery system. Background Art

[0002] Currently, cancer is the biggest killer threatening human health. The conventional methods for cancer treatment clinically include surgical resection, radiotherapy, chemotherapy, or a combination therapy of the above regimens. Among them, chemotherapy is one of the clinically preferred treatment regimens. However, traditional chemotherapy drugs have poor targeting and toxic side effects on normal cells.

[0003] During the occurrence and metastasis of tumors, the relationship between the internal and external environments of tumor cells is the tumor microenvironment (TME). The TME is relatively hypoxic, and the contents of hydrogen peroxide (H2O2) and glutathione (GSH) are relatively high. These factors have hindered the treatment effects of conventional methods to a certain extent.

[0004] Therefore, aiming at the tumor microenvironment, constructing a composite nanozyme system with high enzymatic reaction efficiency for a tumor dual-targeted drug delivery system has more obvious advantages in clinical cancer treatment. Summary of the Invention

[0005] The present disclosure provides an application of a composite nanozyme material in a tumor dual-targeted drug delivery system to at least solve the above technical problems existing in the prior art.

[0006] The present disclosure provides an application of a composite nanozyme material in a tumor dual-targeted drug delivery system. The composite nanozyme material is used in the preparation of an anti-tumor injection or an implantable medicament. The composite nanozyme material has peroxidase-like activity (POD), catalase-like activity (CAT), and glutathione oxidase-like activity (GSH-Ox).

[0007] The application method includes:

[0008] Using the composite nanozyme material as a contrast agent to determine the tumor location by magnetic resonance;

[0009] Setting a magnetic field at the tumor location and injecting an injection or a medicament containing the composite nanozyme material.

[0010] In an implementable embodiment, the composite nanozyme material is prepared by in-situ growing a copper ion-doped metal-organic framework material UiO66-NH2(Cu) on the surface of magnetic nanoparticles composed of multiple metals to form a core-shell structure, then bonding oxidized hyaluronic acid to the surface of the core-shell structure, and then loading a chemotherapeutic drug to obtain a tumor-targeted composite nanozyme material.

[0011] In one feasible embodiment, a magnetic field is set at the tumor location, and the magnetic induction intensity is 3800 - 4200 Gs.

[0012] In one feasible embodiment, for the injection or medicament containing the composite nanozyme material, the injection amount is 15 - 25 mg / Kg.

[0013] In one feasible embodiment, the drug loading amount of the chemotherapeutic drug in the composite nanozyme material is 36.3 - 38.3%.

[0014] In one feasible embodiment, the encapsulation efficiency of the chemotherapeutic drug in the composite nanozyme material is 88.1 - 90.1%.

[0015] In one feasible embodiment, the crystal size of the composite nanozyme is 35 - 45 nm.

[0016] In one feasible embodiment, in the composite nanozyme, the copper ion - doped metal - organic framework material UiO66 - NH2(Cu), Zr 4+ and Cu 2+ have a molar ratio of 1:2.

[0017] The application of a composite nanozyme material disclosed in the present invention in a tumor dual - target drug delivery system. In view of the characteristics of the tumor microenvironment and combining with the novel cell death mode of ferroptosis, the present invention provides a method for the application of a composite nanozyme material in a tumor dual - target drug delivery system. First, a magnetic composite nanozyme material is used as a MIR contrast agent, and the location of the tumor is determined by magnetic resonance technology. Further, the biological activity targeting of the composite nanozyme and the physical magnetic targeting of the external magnetic field of the tumor tissue are utilized to achieve the specific release of the chemotherapeutic drug functional molecules at the target site. The composite nanozyme material is prepared by in - situ growth of a copper ion - doped metal - organic framework material UiO66 - NH2(Cu) on the surface of magnetic nanoparticles composed of multiple metals to form a core - shell structure, then bonding oxidized hyaluronic acid to the surface of the core - shell structure, and then loading chemotherapeutic drugs to obtain a tumor - targeting composite nanozyme material. The present invention utilizes the amino groups on its surface to form acid - sensitive Schiff base bonds with the aldehyde groups of OHA molecules. The nanoparticles modified by OHA stably exist in the physiological environment, while the Schiff base bonds break in the acidic environment of the tumor tissue, releasing the nanoparticles. The higher H2O2 in the tumor microenvironment is used to generate oxygen to improve tissue hypoxia; H2O2 is catalyzed into toxic •OH to induce apoptosis of tumor cells; the high concentration of GSH in the tumor microenvironment is consumed, causing the inactivation of GPX4, resulting in ferroptosis. Through a series of cascade enzyme activity reactions, the maximum killing effect on tumors is achieved.

[0018] It should be understood that the content described in this part is not intended to identify the key or important features of the embodiments of the present disclosure, nor is it used to limit the scope of the present disclosure. Other features of the present disclosure will become easily understood through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] By referring to the following detailed description with reference to the accompanying drawings, the above and other objects, features, and advantages of the exemplary embodiments of the present disclosure will become readily understandable. In the drawings, several embodiments of the present disclosure are shown in an exemplary but non-limiting manner, wherein:

[0020] In the drawings, the same or corresponding reference numerals denote the same or corresponding parts.

[0021] Figure 1 The scanning electron microscope (SEM) image of the zinc ferrite nanomaterial (ZnFe2O4) prepared in the embodiment of the present disclosure is shown;

[0022] Figure 2 The XRD pattern of the zinc ferrite nanomaterial (ZnFe2O4) prepared in the embodiment of the present disclosure is shown;

[0023] Figure 3 The scanning electron microscope (SEM) image of UiO66-NH2(Cu) prepared in the embodiment of the present disclosure is shown;

[0024] Figure 4 The Zr 4+ and Cu 2+ The XRD patterns of UiO66-NH2(Cu) with different ratios are shown;

[0025] Figure 5 The Zr 4+ and Cu 2+ The CAT activities of UiO66-NH2(Cu) with different ratios are shown;

[0026] Figure 6 The Fourier transform infrared spectrum (FTIR) of OHA prepared in the embodiment of the present disclosure is shown;

[0027] Figure 7 The scanning electron microscope (SEM) image of the tumor-targeted composite nanozyme material prepared in the embodiment of the present disclosure is shown;

[0028] Figure 8 The XRD pattern of the tumor-targeted composite nanozyme material prepared in the embodiment of the present disclosure is shown;

[0029] Figure 9 The Fourier transform infrared spectrum (FTIR) of the tumor-targeted composite nanozyme material prepared in the embodiment of the present disclosure is shown;

[0030] Figure 10 The ultraviolet absorption spectrum of the tumor-targeted composite nanozyme material prepared in the embodiment of the present disclosure is shown;

[0031] Figure 11 The enzymatic kinetic curve of the tumor-targeted composite nanozyme material prepared in the embodiments of the present disclosure with H2O2 as the substrate is shown;

[0032] Figure 12 The consumption diagram of H2O2 by the tumor-targeted composite nanozyme material prepared in the embodiments of the present disclosure is shown;

[0033] Figure 13 The curve of dissolved oxygen generated by catalyzing H2O2 by each material prepared in the embodiments of the present disclosure under different pH conditions is shown;

[0034] Figure 14 The consumption of GSH by different concentrations of D / Z@UCO prepared in the embodiments of the present disclosure is shown;

[0035] Figure 15 The T2-weighted images and signal intensities at different concentrations of D / Z@UCO prepared in the embodiments of the present disclosure are shown;

[0036] Figure 16 The magnetic picture of D / Z@UCO prepared in the embodiments of the present disclosure is shown (left: without magnet, right: with magnet);

[0037] Figure 17 The schematic diagram of cell viability measured after co-incubating D / Z@UCO prepared in the embodiments of the present disclosure with mouse breast cancer 4T1 cells for 24 h is shown;

[0038] Figure 18 The killing effect on tumor cells of D / Z@UCO prepared in the embodiments of the present disclosure under different pH and different H2O2 concentration conditions is shown;

[0039] Figure 19 The confocal laser scanning microscope (CLSM) image of the change in intracellular O2 level of D / Z@UCO prepared in the embodiments of the present disclosure is shown;

[0040] Figure 20 The statistical result of the average fluorescence intensity of the intracellular O2 level of D / Z@UCO prepared in the embodiments of the present disclosure is shown;

[0041] Figure 21 The intracellular GSH level of D / Z@UCO prepared in the embodiments of the present disclosure is shown;

[0042] Figure 22 The CLSM image of the cell expressing GPX4 after the action of D / Z@UCO is shown;

[0043] Figure 23 The average fluorescence statistics of the CLSM image of the cell expressing GPX4 after the action of D / Z@UCO are shown;

[0044] Figure 24 Show the cell uptake images after D / Z@UC, D / Z@UCO, and D / Z@UCO+M treatments;

[0045] Figure 25 Show the weights and pictures of tumors after 21 days in different treatment groups. Detailed implementation manners

[0046] To make the objectives, features, and advantages of the present disclosure more obvious and understandable, the technical solutions in the embodiments of the present disclosure will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present disclosure. Obviously, the described embodiments are only a part of the embodiments of the present disclosure, rather than all the embodiments. Based on the embodiments in the present disclosure, all other embodiments obtained by those skilled in the art without creative efforts belong to the scope of protection of the present disclosure.

[0047] Application of a composite nanozyme material in a tumor dual-targeted drug delivery system, the composite nanozyme material is used in the preparation of an anti-tumor injection or implantable medicament, and the composite nanozyme material has peroxidase-like activity (POD), catalase-like activity (CAT), and glutathione oxidase-like activity (GSH-Ox);

[0048] The application method includes:

[0049] S1. Using the composite nanozyme material as a contrast agent, determining the tumor location by magnetic resonance;

[0050] S2. Setting a magnetic field at the tumor location and injecting an injection or medicament containing the composite nanozyme material. The magnetic induction intensity of the magnetic field is 3800 - 4200 Gs.

[0051] Determine the injection volume according to the body weight of the subject. The injection volume is 15 - 25 mg / Kg. Assuming the injection volume is 20 mg / Kg, for example, for a 60 kg subject, the injection volume is 1200 mg.

[0052] In one example, the composite nanozyme material is a core-shell structure formed by in-situ growth of copper ion-doped metal-organic framework material UiO66-NH2(Cu) on the surface of magnetic nanoparticles composed of multiple metals, and then oxidizing hyaluronic acid is bonded to the surface of the core-shell structure, and then a chemotherapy drug is loaded to prepare a tumor-targeted composite nanozyme material.

[0053] The preparation method of the composite nanozyme material includes the following steps:

[0054] 1. Taking magnetic nanoparticles composed of multiple metals as an example of zinc ferrite nanozyme, the preparation of zinc ferrite nanozyme (ZnFe2O4):

[0055] Dissolve 0.164 g of zinc chloride, 0.649 g of ferric chloride hexahydrate, 0.240 g of sodium citrate dihydrate and 1.200 g of sodium acetate in diethylene glycol solution. Ultrasonically stir the above mixture at room temperature, then transfer the obtained suspension into a reaction kettle and heat it at 200 °C for 12 h; wash the obtained product successively with pure water and absolute ethanol, centrifuge it at 11000 r / min for 20 min, and dry it in vacuum at 60 °C for 12 h to obtain ZnFe2O4 (Z) nanomaterials.

[0056] 1.1 Characterization of ZnFe2O4

[0057] Obtain the morphology of zinc ferrite nanomaterials by scanning electron microscopy (SEM). As Figure 1 shown, the obtained ZnFe2O4 is in a spherical structure with a particle size of about 30 nm. Use an X-ray powder diffractometer (XRD) to analyze and verify its crystal structure. The results are as Figure 2 shown. The obtained ZnFe2O4 XRD pattern corresponds to the ZnFe2O4 standard card (JCPDS 22-1012), indicating that ZnFe2O4 has been successfully prepared.

[0058] 2. Prepare ZnFe2O4@UiO66-NH2(Cu) (Z@UC) powder, that is, in-situ grow copper ion-doped metal-organic framework material UiO66-NH2(Cu) on the surface of zinc ferrite nanomaterials;

[0059] 2.1. First, prepare UiO66-NH2(Cu) separately in the present invention. The preparation process is as follows:

[0060] Dissolve 0.0814 g of zirconium chloride and 0.1187 g of copper chloride in 10 mL of N,N-dimethylformamide (DMF) to obtain solution A; dissolve 2-aminoterephthalic acid in 10 mL of DMF to obtain solution B; mix solution A, solution B and 0.6 mL of acetic acid evenly in a beaker, ultrasonically treat for 5 min, pour it into a polytetrafluoroethylene reaction kettle, and place it in an oven at 100 °C for 24 h. After cooling to room temperature, obtain suspension C; centrifuge suspension C at 11000 r / min for 5 min, discard the supernatant to obtain precipitate D, wash precipitate D with DMF three times, then wash precipitate D with absolute ethanol three times, and finally dry it overnight in a vacuum oven at 60 °C to obtain UiO66-NH2(Cu) (UC) powder.

[0061] 2.2 Characterization of UiO66-NH2(Cu)

[0062] The SEM image of UiO66-NH2(Cu) nanoparticles is asFigure 3 As shown, the UiO66-NH2(Cu) nanoparticles are octahedrons with a crystal size of approximately 100 nm. The XRD pattern is as Figure 4 shown. The XRD characterization peaks are consistent with the crystal structure of the nanoparticles reported in the literature, confirming the successful synthesis of the UiO66-NH2(Cu) nanoparticles. In addition, the effects of the ratios of Zr 4+ and Cu 2+ in UiO66-NH2(Cu) on the structure and enzyme activity were investigated. Since the lattice of Cu 2+ is smaller than that of Zr 4+ , the XRD peaks will shift slightly to the right. As shown in Figure 4 , as the ratio of Cu 2+ increases, the characteristic peaks shift more to the right. In addition, due to the doping of Cu 2+ affecting its enzyme activity, considering the importance of both structural stability and enzyme activity factors, the UiO66-NH2(Cu) prepared with a Zr 4+ :Cu 2+ ratio of 1:2 was finally determined for subsequent experiments. As shown in Figure 5 , the CAT activity was investigated using the titanium sulfate colorimetric method. After adding 50 μL of H2O2 (150 mM) and 200 μL of UiO66-NH2(Cu) (1 mg / mL) to the buffer and mixing for 30 min, it was added to the titanium sulfate solution. After 10 min, the concentration of H2O2 was obtained by measuring the absorbance at 410 nm. The stronger the CAT activity, the less H2O2 remained and the lighter the color. As shown in Figure 5 , as the ratio of Cu doping 2+ increases, the color gradually becomes lighter and the CAT activity is higher.

[0063] 2.3. Preparation of ZnFe2O4@UiO66-NH2(Cu) (Z@UC) powder

[0064] 80 mg of ZnFe2O4 was dispersed in 0.6 mM 3-mercaptopropionic acid (3-MPA) solution (20 mL), magnetically stirred for 24 h, and then washed with ultrapure water and ethanol. 0.414 mmol, 80 mg of ZnFe2O4 was added to 10 mL of DMF and sonicated for 20 min. Zirconium chloride (0.3493 mmol, 0.0814 g) was added and stirred for 30 min, then copper chloride (0.0874 mmol, 0.0140 g) was added and stirred for 30 min. Acetic acid (0.6 mL) was added and stirred for 3 min, and then 2-aminoterephthalic acid (0.3493 mmol, 0.0621 g) was added and stirred for 1 h. Finally, the reaction mixture was poured into a polytetrafluoroethylene autoclave, sealed and reacted at 100 °C for 12 h. The final product was collected by centrifugation, rinsed with DMF / absolute ethanol and dried in vacuo at 70 °C to obtain ZnFe2O4@UiO66-NH2(Cu) (Z@UC) powder.

[0065] To further improve the dispersion effect of ZnFe2O4@UiO66-NH2(Cu) (Z@UC) powder, 10 mg of Z@UC powder was further added to 5 mL of ultrapure water and sonicated for 1 h. 100 mg of sodium citrate (Na3Cit·2H2O) was weighed and dissolved in 15 mL of ultrapure water. The ZnFe2O4@UiO66-NH2(Cu) dispersion was added dropwise to the Na3Cit·2H2O aqueous solution and reacted at 80 °C for 30 min. After cooling to room temperature, it was stirred for 12 h. It was collected by centrifugation to obtain ZnFe2O4@UiO66-NH2(Cu)-Cit.

[0066] 3. Preparation of oxidized hyaluronic acid (OHA)

[0067] 0.5 g of hyaluronic acid (HA) was dissolved in 50 mL of ultrapure water, and 8 mL of 3.3% sodium periodate (NaIO4) aqueous solution was added to make the molar ratio of HA to NaIO4 1:1. It was stirred at room temperature in the dark for 12 h, dialyzed for 3 days and freeze-dried to obtain OHA. The reaction formula is as follows:

[0068]

[0069] 3.1 Characterization of OHA

[0070] Figure 6 The Fourier transform infrared spectroscopy (FTIR) of the OHA sample prepared in this example is shown. According to Figure 6 It can be seen that all HA and OHA have polysaccharide characteristics. The presence of aldehyde groups in OHA can be determined by 1730 cm -1The sharp peak (-C=O stretching vibration) is used to determine the successful synthesis of OHA.

[0071] 4. Preparation of tumor-targeted composite nanozyme material DOX / ZnFe2O4@UiO66-NH2(Cu)-OHA (D / Z@UCO)

[0072] In this example, doxorubicin (DOX) is used as the chemotherapeutic drug. Dissolve 10 mg of OHA in 20 mL of PBS solution. Disperse 10 mg of ZnFe2O4@UiO66-NH2(Cu)-Cit and 10 mg of doxorubicin (DOX) in PBS (pH = 7.4) solution and ultrasonically mix for 1 h. Then, slowly add the mixture dropwise to the OHA solution. Adjust the pH of the mixed solution to 12 and react for 2 h. Then, adjust the pH back to 7.4 and stir in the dark for 12 h. Centrifuge to collect the final tumor-targeted composite nanozyme material DOX / ZnFe2O4@UiO66-NH2(Cu)-OHA (D / Z@UCO).

[0073] The drug loading (DL) and encapsulation efficiency (EE) are measured using a UV spectrophotometer and calculated as follows:

[0074] DL% = (W0 - C f V f ) / W0 × 100% Formula (1)

[0075] EE% = (W0 - C f V f ) / C p V p × 100% Formula (2)

[0076] In the formula, W0 is the dosage of the drug, C f , V f are the concentration and volume of free DOX, and C p , V p are the concentration and volume of the D / Z@UCO dispersion.

[0077] After calculation, the drug loading is 37.3 ± 1%, and the encapsulation amount is 89.1 ± 1%

[0078] 4.1 Characterization of D / Z@UCO

[0079] The SEM image of D / Z@UCO is as Figure 7 shown. Morphological observation shows that D / Z@UCO is spherical, and the crystal size is about 40 nm; Figure 8 The XRD crystal structure shown indicates that the characteristic peaks match the crystal structure of the single-component nanoparticles, confirming the successful synthesis of D / Z@UCO; FTIR is as Figure 9As shown, compared with individual components, the composite material has the characteristic peaks of all individual components, indicating the successful synthesis of D / Z@UCO.

[0080] 4.2 Enzymatic activity characterization of D / Z@UCO

[0081] The POD activity of the above-prepared D / Z@UCO was determined by the color reaction of 3,3',5,5'-tetramethylbenzidine (TMB). That is, composite nanozyme materials with different concentrations were added to 0.1 M acetic acid-sodium acetate buffer solutions with different pH values. The final concentration of H2O2 was 0.5 M, and the final concentration of TMB was 0.4 mM. After mixing, the absorbance of the solution at 652 nm was measured at different time points.

[0082] As Figure 10 shown by the absorbance results, as the concentration of D / Z@UCO increases, the production amount of oxidized TMB is more, indicating higher peroxidase (POD) activity. As Figure 11 shown by the enzymatic kinetic curve with H2O2 as the substrate, it was measured from the D / Z@UCO nanozyme dispersion in H2O2 solutions with different concentrations, where the final concentration of TMB was 0.4 mM. From Figure 11 it can be obtained that the affinity constant Km of the prepared D / Z@UCO is 1.061 μM and the maximum reaction rate Vmax is 139.4 nMs -1 .

[0083] By measuring the consumption and oxygen production levels of D / Z@UCO for H2O2, it was proved that the above-prepared D / Z@UCO also has CAT activity. As Figure 12 shown, as the concentration of D / Z@UCO increases, the consumption of H2O2 gradually increases, indicating higher CAT activity. Figure 13 Shown is the schematic diagram of the curve of D / Z@UCO catalyzing H2O2 to produce dissolved oxygen under different pH conditions. D / Z@UCO produces the highest amount of O2 in an environment with a pH of 6.5, indicating that D / Z@UCO can effectively produce O2 under the weakly acidic conditions at the tumor site, relieve hypoxia at the tumor site, and thus improve the killing effect on tumors.

[0084] By measuring the consumption of GSH by D / Z@UCO, it was proved that the above-prepared D / Z@UCO also has GSH-Ox enzyme activity. As Figure 14 shown, as the concentration of D / Z@UCO increases, the consumption of GSH is faster, indicating better GSH-Ox activity.

[0085] It was further confirmed that this composite nanozyme material can be used as a MIR contrast agent. The application method is as follows:

[0086] Disperse D / Z@UCO in 2% agarose gel at concentrations ranging from 0 to 1000 mg / mL. Use a 3.0 T nuclear magnetic resonance spectrometer from GE Company, as Figure 15 shown in the T2-weighted images and signal intensities at different D / Z@UCO concentrations; Figure 16 are the magnetic pictures of D / Z@UCO (left: without magnet, right: with magnet). According to Figure 15 and Figure 16 The results show that D / Z@UCO has certain magnetism, and the T2-weighted images gradually darken with the increase in D / Z@UCO concentration, indicating that D / Z@UCO can be used as a T2 contrast agent.

[0087] 5. Antitumor effect of composite nanozyme materials at the cellular level

[0088] The above-prepared D / Z@UCO was co-incubated with mouse breast cancer 4T1 cells for 24 h under normal oxygen concentration and hypoxic conditions respectively, and the cell viability was measured by the MTT method. The results are as Figure 17 shown. With the increase in D / Z@UCO concentration, the cell viability gradually decreases. Under hypoxic conditions, D / Z@UCO produces oxygen through CAT activity, and three enzyme activity cascades to kill tumor cells. Figure 18 Shown is the killing effect of the nanoparticles on tumor cells with the increase in H2O2 concentration under different pH and different H2O2 concentration conditions, and it is more beneficial for D / Z@UCO to play a role under acidic conditions. Due to the GSH-Ox activity of D / Z@UCO, the concentration of GSH is reduced, the consumption of •OH is reduced, the activity of GPX4 is reduced, ferroptosis is induced, and with the help of CAT activity, the hypoxic conditions inside the tumor are alleviated, which is beneficial for the efficacy of DOX and the accumulation of a large amount of •OH in cancer cells, greatly improving the killing effect of the nanoparticles on tumors.

[0089] By measuring the oxygen content in cells with an RDPP hypoxia probe, it is proved that the nanomaterial alleviates the hypoxia in the tumor microenvironment, and the results are recorded by CLSM. As Figure 19 and Figure 20 shown, with the increase in H2O2 concentration, the intracellular oxygen level gradually increases, indicating that the nanomaterial can effectively produce O2 and alleviate the hypoxia inside the tumor.

[0090] The change in the intracellular GSH level of 4T1 cells was further studied as Figure 21 shown. The GSH-Ox activity of D / Z@UCO reduces the intracellular GSH level, leading to the inactivation of GPX4 and ultimately resulting in ferroptosis. As Figure 22 and Figure 23As shown, with the increase in the concentration of D / Z@UCO, the intracellular GSH concentration decreased, indicating that D / Z@UCO has good GSH-Ox activity in cells. The expression of GPX4 in 4T1 cells was studied by CLSM. With the increase in the concentration of D / Z@UCO, the intracellular GPX4 fluorescence intensity gradually decreased, indicating that D / Z@UCO can induce ferroptosis in cells.

[0091] To prove the targeting of D / Z@UCO, the dual-targeting effect was demonstrated through a cell uptake experiment. Utilizing the fluorescence characteristics of DOX, the cell uptake of each nanoparticle by D / Z@UC (non-targeting group), D / Z@UCO (OHA targeting group), and D / Z@UCO+M (OHA targeting group and magnetic targeting group) was observed at 2 h and 4 h, as Figure 24 shown. There was basically no drug enrichment in the non-targeting group, while there was a small amount of drug enrichment around the tumor cells in the single OHA targeting group. After applying the magnetic field, the drug enrichment was significantly enhanced, indicating that the dual-targeting effect of the OHA and magnetic field groups showed the best cell uptake effect.

[0092] 6. Antitumor effect of the composite nanomaterial enzyme at the animal level

[0093] A mouse orthotopic breast cancer model was constructed. When the tumor reached 50 mm 3 , treatment began. The mice were randomly divided into 5 groups, namely the PBS group, Z@UCO (no drug added, non-magnetic targeting group), Z@UCO+M (no drug added, magnetic targeting group), D / Z@UCO (drug added, non-magnetic targeting group), and D / Z@UCO+M (drug added, magnetic targeting group). The tumor-targeting composite nanozyme material prepared above was diluted to a certain concentration and injected into the tumor-bearing mice via the tail vein, and the tumor size of the tumor-bearing mice was statistically analyzed. The results are as Figure 25 shown. Compared with the control group, the tumors of the experimental mice in the Z@UCO, Z@UCO+M, D / Z@UCO, and D / Z@UCO+M groups were all inhibited to a certain extent. Compared with the non-magnetic targeting groups of Z@UCO and D / Z@UCO, the tumor inhibition effect of the magnetic targeting groups of Z@UCO+M and D / Z@UCO+M on the experimental mice was stronger, and the tumors of the mice in the D / Z@UCO+M drug-added group showed an obvious shrinking trend.

[0094] When the composite nanomaterial enzyme material is applied clinically, first, the composite nanomaterial enzyme material is used as a contrast agent to determine the tumor location by magnetic resonance, then a magnetic field is set at the tumor location with a magnetic induction intensity of 4000 Gs, and then an injection or dosage containing 20 mg / Kg of the composite nanomaterial enzyme is injected.

[0095] It should be understood that various forms of the processes shown above can be used, with steps reordered, added, or deleted. For example, the steps described in this disclosure can be executed in parallel, sequentially, or in a different order, as long as the desired results of the technical solution disclosed in this disclosure can be achieved, and this is not limited herein.

[0096] In addition, the terms "first" and "second" are used only for descriptive purposes and cannot be construed as indicating or implying relative importance or implicitly specifying the quantity of the indicated technical features. Thus, features defined with "first" and "second" may explicitly or implicitly include at least one of such features. In the description of this disclosure, "a plurality" means two or more, unless otherwise specifically defined.

[0097] As described above, the above are only specific embodiments of this disclosure, but the protection scope of this disclosure is not limited thereto. Any person skilled in the art within the technical scope disclosed in this disclosure can easily think of changes or substitutions, which should all be covered within the protection scope of this disclosure. Therefore, the protection scope of this disclosure should be subject to the protection scope of the claims.

Claims

1. Use of a composite nanozyme material in the preparation of an anti-tumor injection or implantable medicament, characterized in that the composite nanozyme material has peroxidase-like activity, catalase-like activity and glutathione oxidase-like activity; the composite nanozyme material is formed by in-situ growth of copper ion-doped metal-organic framework material UiO66-NH2(Cu) on the surface of magnetic nanoparticles composed of ZnFe2O4 to form a core-shell structure, then bonding oxidized hyaluronic acid to the surface of the core-shell structure, and then loading chemotherapy drugs to obtain a tumor-targeting composite nanozyme material.

2. The application according to claim 1, wherein The drug loading amount of the chemotherapy drug in the composite nanozyme material is 36.3-38.3%.

3. The application according to claim 1, wherein The encapsulation efficiency of the chemotherapy drug in the composite nanozyme material is 88.1-90.1%.

4. The application according to claim 1, characterized in that, The crystal size of the composite nanozyme is 35-45 nm.

5. The application according to claim 1, wherein In the composite nanozyme, the copper ion-doped metal-organic framework material UiO66-NH2(Cu), Zr 4+ and Cu 2+ have a molar ratio of 1:2.

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