A raspberry-like basic copper chloride nanomaterial with dual enzymatic activity, its preparation method and application

Abstract

A raspberry-like basic copper chloride nanomaterial with dual enzymatic activity, its preparation method, and its application are disclosed. The steps are as follows: Dimethyl sulfoxide is added to an aqueous sodium hydroxide solution containing N-fluorenemethoxycarbonyl-L-arginine (Fmoc-L-Arg), and the solution is sonicated until clear to obtain mixed solution A. An aqueous solution of copper chloride dihydrate (CuCl2·2H2O) is prepared for later use. The CuCl2·2H2O aqueous solution is added to mixed solution A to obtain mixed solution B, and the reaction is allowed to stand. After the reaction is complete, the precipitate is collected by centrifugation and washed with water. This invention employs a biomimetic mineralization method, using Fmoc-L-Arg molecules as templates to guide and control the formation of uniformly sized (130 nm) Cu2(OH)3Cl nanomaterials. These nanomaterials simultaneously possess dual enzymatic activities similar to peroxidase and glutathione oxidase, exhibiting good biocompatibility and enabling specific killing of tumor cells.

Description

Technical Field

[0001] This invention relates to the field of biomedical nanomaterials technology, specifically to a raspberry-like basic copper chloride nanomaterial with dual enzymatic activity, its preparation method, and its application. Background Technology

[0002] Transition metal nanomaterials have shown great promise in the field of tumor therapy, especially in chemokinetic therapy. Among them, copper-based nanomaterials, after being internalized by tumor cells, release copper ions that can not only react with excess hydrogen peroxide in the tumor microenvironment in a Fenton-like reaction to generate highly toxic hydroxyl radicals, but also consume excess glutathione, thereby efficiently and specifically killing tumor cells.

[0003] Basic copper chloride (Cu₂(OH)₃Cl) is characterized by good stability and high copper content, exhibiting superior bioavailability and biosafety compared to other copper-based materials. As a promising copper-based nanomaterial, its conventional synthesis methods mainly include hydrothermal and solvothermal methods. However, these methods often require harsh conditions, operating under high temperature and pressure, and are time-consuming (typically several to tens of hours), with complex processes and high energy consumption, severely restricting the further development and application of this material. Furthermore, traditional synthetic routes primarily focus on the mineral properties of Cu₂(OH)₃Cl, with less attention paid to its morphology control, especially regarding the limited reports on nanoscale morphology regulation of Cu₂(OH)₃Cl. This results in insufficient exposure of the material's catalytic active sites, thus severely limiting its catalytic efficiency.

[0004] Based on this, this invention employs a biomimetic mineralization method. Under mild reaction conditions, an organic template is constructed using N-fluorenylmethoxycarbonyl-L-arginine (Fmoc-L-Arg) molecules. Through the coordination of Fmoc-L-Arg with copper chloride dihydrate (CuCl2·2H2O), raspberry-like Cu2(OH)3Cl nanomaterials with a size of only 130 nm are precisely induced. Using 3,3',5,5'-tetramethylbenzidine (TMB) as a probe, the highly efficient Fenton-like catalytic activity of the Cu2(OH)3Cl nanomaterials was demonstrated. Using 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) as a chromogenic reagent, the glutathione oxidase-like activity of the Cu2(OH)3Cl nanomaterials was characterized. Further cytotoxicity and live / dead staining studies showed that the Cu2(OH)3Cl nanomaterials possess good biocompatibility. Simultaneously, it consumes excess glutathione within cells to undermine their antioxidant defenses; on the other hand, it enhances copper-mediated Fenton-like reactions to accelerate the production of reactive oxygen species. This dual mechanism works synergistically to ultimately and efficiently induce tumor cell death. Summary of the Invention

[0005] Technical Problem Solved: Addressing the problems of demanding synthesis conditions, lengthy processing times, and difficulties in size control in existing Cu2(OH)3Cl synthesis techniques, this invention provides a raspberry-like basic copper chloride nanomaterial with dual enzymatic activity, its preparation method, and its applications. Employing a biomimetic mineralization method under mild reaction conditions, using Fmoc-L-Arg molecules as organic templates, and leveraging their coordination with CuCl2·2H2O, the formation of raspberry-like Cu2(OH)3Cl nanomaterials with a size of only 130 nm is precisely guided, thus developing a mild preparation method for Cu2(OH)3Cl nanomaterials. Furthermore, this invention verifies the highly efficient peroxidase-like and glutathione oxidase-like dual catalytic activities of the Cu2(OH)3Cl nanomaterials, ultimately confirming their excellent killing effect on tumor cells.

[0006] Technical solution: The first objective of this invention is to provide a method for preparing raspberry-like basic copper chloride nanomaterials with dual enzymatic activity, the steps of which are as follows:

[0007] Step 1: Add sodium hydroxide aqueous solution to N-fluorenylmethoxycarbonyl-L-arginine (Fmoc-L-Arg), mix by sonication, then add dimethyl sulfoxide to obtain a mixture. Sonicate the mixture until the solution is clear to obtain mixed solution A; prepare CuCl2·2H2O aqueous solution for later use.

[0008] Step 2: Quickly add CuCl2·2H2O aqueous solution to mixed solution A within one second to obtain mixed solution B, and let it stand to react;

[0009] Step 3: After the reaction is complete, the precipitate is collected by centrifugation and washed with water to obtain the raspberry-like basic copper chloride nanomaterial with dual enzyme activity, denoted as Cu2(OH)3Cl nanomaterial.

[0010] Preferably, in step one, the ratio of Fmoc-L-Arg, sodium hydroxide aqueous solution, and dimethyl sulfoxide in mixed solution A is 90-110 mg: 200-250 μL: 4-4.5 mL, and the concentration of the added sodium hydroxide aqueous solution is 0.115 M.

[0011] Preferably, in step one, the concentration of the CuCl2·2H2O aqueous solution is 0.16-0.17M.

[0012] Preferably, in step two, the volume ratio of CuCl2·2H2O aqueous solution to mixed solution A is 1.35-1.5:1.

[0013] Preferably, in step two, the reaction conditions are: reacting at 24-28°C for 5-8 minutes.

[0014] Preferably, in step three, the centrifugation conditions are: centrifugation at 6000-9000 rpm for 5 min.

[0015] Preferably, in step three, the washing with water is performed as follows: wash with water 1-3 times, and then disperse the washed nanomaterials in ultrapure water for later use.

[0016] The second objective of this invention is to provide a raspberry-like basic copper chloride nanomaterial with dual enzymatic activity prepared based on the above method.

[0017] A third objective of this invention is to provide the application of a raspberry-like basic copper chloride nanomaterial with dual enzymatic activity, as described above, in the preparation of a drug for tumor treatment.

[0018] Beneficial effects:

[0019] (1) The preparation method of Cu2(OH)3Cl nanomaterials provided by the present invention is mild and simple. The reaction does not require high temperature, high pressure and complex equipment, but only needs to be completed by short-term standing at room temperature and pressure, and the product does not require complicated subsequent purification treatment.

[0020] (2) The method for preparing Cu2(OH)3Cl nanomaterials provided by this invention can obtain uniform raspberry-like nanomaterials with a size of 130 nm. Compared with the amorphous precipitate structure that is easily obtained by traditional hydrothermal and solvothermal methods, this unique small-sized microstructure has fully exposed active sites and has higher peroxidase-like and glutathione oxidase-like catalytic activities.

[0021] (3) The preparation method of Cu2(OH)3Cl nanomaterials provided by the present invention uses biomolecules as templates and surface modification units, and has good biocompatibility. Attached Figure Description

[0022] Figure 1 TEM (A), SEM (B), and EDX mapping (C) images of the Cu2(OH)3Cl nanomaterials prepared in Example 1.

[0023] Figure 2 X-ray diffraction pattern (A) and X-ray photoelectron spectrum (B) of Cu2(OH)3Cl nanomaterials prepared in Example 1.

[0024] Figure 3The figure shows the catalytic performance test results of Cu2(OH)3Cl nanomaterial in Application Example 1. In the figure, A is the UV spectrum of Cu2(OH)3Cl nanomaterial catalyzing the oxidation of TMB (0.5 mM); B is the EPR spectrum of Cu2(OH)3Cl nanomaterial; C is the UV spectrum of Cu2(OH)3Cl nanomaterial after reacting with GSH (0.5 mM) and then adding DTNB; D is a comparison of the changes in the absorbance of Cu2(OH)3Cl nanomaterial for degrading MB over time before and after reacting with GSH.

[0025] Figure 4 Example 1 shows the cytotoxicity evaluation of Cu2(OH)3Cl nanomaterials. In the figure, A is the cell survival rate of L-929 cells and HeLa cells after incubation with different mass concentrations of Cu2(OH)3Cl nanomaterials for 24 h; B is the fluorescence imaging of HeLa cells before and after treatment with Cu2(OH)3Cl nanomaterials, after AM / PI co-staining. Detailed Implementation

[0026] The present invention will be further described below with reference to the accompanying drawings and specific embodiments.

[0027] Unless otherwise specified, the raw materials used in the embodiments of this invention are all from commercially available products, including:

[0028] N-fluorenylmethoxycarbonyl-L-arginine was purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China).

[0029] Copper chloride dihydrate and methylene blue (MB) were purchased from Shanghai Maclean Biotechnology Co., Ltd. (Shanghai, China).

[0030] Dimethyl sulfoxide and sodium hydroxide were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai).

[0031] Glutathione (GSH) and 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) were purchased from Shanghai Aladdin Biochemical Co., Ltd. (Shanghai, China).

[0032] The Cell Proliferation-Cytotoxicity Assay Kit (CCK-8) and the Calcein / PI Cell Viability and Cytotoxicity Assay Kit were purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China).

[0033] The buffer solution was PBS, which was purchased from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China).

[0034] Example 1

[0035] This embodiment provides a method for preparing Cu2(OH)3Cl nanomaterials with dual enzymatic activity, as detailed below:

[0036] 200 μL of sodium hydroxide aqueous solution (0.115 M) was added to 100 mg (0.25 mmol) Fmoc-L-Arg. After simple sonication and mixing, 4 mL of dimethyl sulfoxide was added to obtain a mixture. The mixture was sonicated until the solution became clear to obtain mixed solution A. Then, 5.8 mL of CuCl2·2H2O aqueous solution (5.8 mL, 0.165 M) was rapidly added to it within one second to obtain mixed solution B. The mixture was allowed to stand at room temperature (25 °C) for 5 minutes under normal temperature and pressure conditions to form Cu2(OH)3Cl nanomaterials. After the reaction was completed, the precipitate was collected by centrifugation (9000 rpm, 5 minutes) and washed three times with water to obtain raspberry-like basic copper chloride nanomaterials with dual enzymatic activity, denoted as Cu2(OH)3Cl nanomaterials. The obtained Cu2(OH)3Cl nanomaterials were stored in ultrapure water for later use.

[0037] Depend on Figure 1 (AC) It is known that Cu2(OH)3Cl nanomaterials are raspberry-like nanoparticles with an average diameter of about 130 nm. Cu, Cl, and O elements are uniformly distributed within the nanoparticles, while C and N elements are present in small amounts. This indicates that after deprotonation by sodium hydroxide solution, the unique amphiphilic characteristics of Fmoc-L-Arg molecules facilitate their assembly in solution through hydrophilic-hydrophobic interactions, forming an assembly with an arginine structure pointing towards water and an Fmoc aromatic structure encapsulated within. With the introduction of CuCl2·2H2O aqueous solution, the exposed -COO on the surface of the assembly... - With Cu 2+ Cu combines through coordination. 2+ The formation of the -Fmoc-L-Arg complex is OH - and Cl - Through stronger ionic bonds with Cu 2+ The organic template provides nucleation sites, while its relatively weak coordination is replaced as Cu2(OH)3Cl continues to grow and mature. Ultimately, after the reaction, the organic template molecules largely detach, forming a raspberry-like Cu2(OH)3Cl nanomaterial dominated by ionic bonds. However, due to... Figure 1 It can be seen that C and N elements are also distributed on the surface of nanoparticles, indicating that Fmoc-L-Arg is retained to a certain extent on the surface of Cu2(OH)3Cl nanomaterials, which provides a guarantee for the biocompatibility of Cu2(OH)3Cl nanomaterials.

[0038] Depend on Figure 2 The X-ray diffraction analysis diagram in (A) shows that the diffraction peaks of Cu2(OH)3Cl nanomaterials are consistent with the diffraction peaks of Cu2(OH)3Cl crystals in the PDF standard card. Figure 2 The high-resolution XPS spectrum of Cu2p in (B) shows that Cu 2+ The characteristic binding energies of Cu are 934.67 eV and 954.42 eV, and Cu 2+ The satellite peaks confirm that copper is mainly present as Cu. 2+ It exists in the form of Cu2(OH)3Cl nanomaterials.

[0039] Example 2

[0040] This embodiment provides a method for preparing Cu2(OH)3Cl nanomaterials with dual enzymatic activity, as detailed below:

[0041] Step 1: Add sodium hydroxide aqueous solution to Fmoc-L-Arg, mix by simple sonication, then add dimethyl sulfoxide solution to obtain a mixture. Sonicate the mixture until the solution is clear to obtain mixed solution A. Dissolve CuCl2·2H2O in ultrapure water to obtain CuCl2·2H2O aqueous solution for later use.

[0042] Step 2: Quickly add CuCl2·2H2O aqueous solution to mixed solution A within one second to obtain mixed solution B, and let it stand to react;

[0043] Step 3: After the reaction is complete, the precipitate is collected by centrifugation, washed with water, and then a raspberry-like basic copper chloride nanomaterial with dual enzyme activity is obtained, denoted as Cu2(OH)3Cl nanomaterial.

[0044] In step one, the ratio of Fmoc-L-Arg, sodium hydroxide aqueous solution, and dimethyl sulfoxide in mixed solution A is 90 mg: 250 μL: 4 mL, and the concentration of the added sodium hydroxide aqueous solution is 0.115 M.

[0045] In step one, the concentration of the CuCl2·2H2O aqueous solution is 0.17 M.

[0046] In step two, the volume ratio of CuCl2·2H2O aqueous solution to mixed solution A is 1.4:1.

[0047] In step two, the temperature for the static reaction is 24 ℃, and the reaction time is 5 min.

[0048] In step three, the centrifugation conditions are: centrifugation at 6000 rpm for 5 minutes, followed by washing with water 1-3 times.

[0049] Example 3

[0050] This embodiment provides a method for preparing Cu2(OH)3Cl nanomaterials with dual enzymatic activity, as detailed below:

[0051] Step 1: Add sodium hydroxide aqueous solution to Fmoc-L-Arg, mix by simple sonication, then add dimethyl sulfoxide solution to obtain a mixture. Sonicate the mixture until the solution is clear to obtain mixed solution A. Dissolve CuCl2·2H2O in ultrapure water to obtain CuCl2·2H2O aqueous solution for later use.

[0052] Step 2: Quickly add CuCl2·2H2O aqueous solution to mixed solution A within one second to obtain mixed solution B, and let it stand to react;

[0053] Step 3: After the reaction is complete, the precipitate is collected by centrifugation, washed with water, and then a raspberry-like basic copper chloride nanomaterial with dual enzyme activity is obtained, denoted as Cu2(OH)3Cl nanomaterial.

[0054] In step one, the ratio of Fmoc-L-Arg, sodium hydroxide aqueous solution, and dimethyl sulfoxide in mixed solution A is 110 mg: 200 μL: 4.5 mL, and the concentration of the added sodium hydroxide aqueous solution is 0.115 M.

[0055] In step one, the concentration of the CuCl2·2H2O aqueous solution is 0.16 M.

[0056] In step two, the volume ratio of CuCl2·2H2O aqueous solution to mixed solution A is 1.35:1.

[0057] In step two, the temperature for the static reaction is 28 ℃, and the reaction time is 8 min.

[0058] In step three, the centrifugation conditions are: centrifugation at 6000 rpm for 5 minutes, followed by washing with water 1-3 times.

[0059] Application Example 1

[0060] The catalytic properties and tumor therapy effects of the Cu2(OH)3Cl nanomaterials prepared in Example 1 were studied as follows:

[0061] (1) The catalytic effect of the prepared nanomaterials was evaluated by monitoring the absorbance change of TMB over time in the presence of H2O2. The total reaction volume was 3 mL, the buffer solution was PBS (pH = 6.5), the reaction temperature was 37 °C, the concentration of Cu2(OH)3Cl nanomaterials was 0.01 mg / mL, the concentration of TMB was 0.5 mM, and the concentration of H2O2 was 50 mM, denoted as Cu2(OH)3Cl + H2O2 + TMB.

[0062] Systems without H₂O₂ or without Cu₂(OH)₃Cl nanomaterials were used as controls, denoted as Cu₂(OH)₃Cl+TMB and H₂O₂+TMB, respectively. The reaction solutions were reacted in cuvettes for 1 minute, followed by UV-Vis analysis. Figure 3 The UV-Vis spectrum of (A) shows that Cu2(OH)3Cl + TMB and H2O2 + TMB solutions do not exhibit obvious absorption peaks at 652 nm. However, for the mixed solution of Cu2(OH)3Cl + H2O2 + TMB, after reacting in a cuvette for 1 minute, a significant absorption enhancement was observed at 652 nm, indicating that Cu2(OH)3Cl can catalyze the generation of free radicals from H2O2 and further oxidize TMB.

[0063] (2) Using DMPO as a scavenging agent, the type of free radicals generated by the prepared nanomaterials was detected by electron paramagnetic resonance (EPR) spectroscopy. The total reaction volume was 1 mL, the solvent was PBS buffer solution (pH = 6.5), the reaction temperature was 25 ℃, the concentration of Cu2(OH)3Cl nanomaterial was 0.01 mg / mL, the concentration of DMPO was 100 mM, and the concentration of H2O2 was 50 mM, denoted as DMPO + H2O2 + Cu2(OH)3Cl. Systems without H2O2 or without Cu2(OH)3Cl nanomaterial were used as controls, denoted as DMPO + Cu2(OH)3Cl and DMPO + H2O2, respectively. After mixing the above three systems in a centrifuge tube for 1 minute, a small amount of liquid was transferred using a capillary tube and placed into the test chamber to collect the spectrum. Figure 3 The EPR spectrum in (B) shows that DMPO + Cu2(OH)3Cl and DMPO + H2O2 solutions did not cause significant signal changes, while DMPO + H2O2 + Cu2(OH)3Cl produced four signal peaks with a signal intensity ratio of 1:2:2:1. This indicates that Cu2(OH)3Cl can decompose H2O2 into hydroxyl radicals, which are captured by DMPO and have peroxidase-like catalytic activity.

[0064] (3) The glutathionein-like catalytic effect of the prepared Cu2(OH)3Cl nanomaterials was evaluated by the DTNB colorimetric method. The total reaction volume was 3 mL, the solvent was PBS buffer solution (pH = 6.5), the reaction temperature was 37 ℃, the concentration of Cu2(OH)3Cl nanomaterials was 0.01 mg / mL, the concentration of DTNB was 0.1 mM, and the concentration of GSH was 0.5 mM, denoted as Cu2(OH)3Cl+DTNB+GSH. Systems with only DTNB, GSH, and DTNB+GSH added were used as controls, denoted as DTNB, GSH, and DTNB+GSH, respectively.

[0065] Depend on Figure 3 The UV absorption spectrum of (C) shows that the DTNB+GSH solution alone exhibits a strong absorption peak at 425 nm after reacting for 10 min. However, the absorption peak at 425 nm of the DTNB+GSH solution treated with Cu2(OH)3Cl nanomaterials for 10 min shows a significant decrease after reacting for 10 min. This indicates that Cu2(OH)3Cl nanomaterials can undergo redox reactions with GSH, achieving the effect of glutathione oxidase-like consumption of GSH.

[0066] (4) The peroxidase activity of the prepared nanomaterials after reaction with GSH was evaluated by MB degradation. The total reaction volume was 3 mL, the solvent was PBS (pH = 6.5), the reaction temperature was 37 °C, the concentration of Cu2(OH)3Cl nanomaterials was 0.01 mg / mL, the concentration of GSH was 0.125 mM, the concentration of MB was 10 μg / mL, and the concentration of H2O2 was 50 mM, denoted as Cu2(OH)3Cl + GSH + H2O2 + MB. The system without GSH treatment was used as a control, denoted as Cu2(OH)3Cl + H2O2 + MB.

[0067] Depend on Figure 3 (D) It can be seen that without GSH treatment for 10 min, the degradation rate of MB by Cu2(OH)3Cl nanomaterials was 63.5% within 10 min, while after GSH treatment for 10 min, the degradation rate of MB reached as high as 94%. The increased peroxidase activity induced by this process is mainly due to the following two points: on the one hand, GSH, as a strong reducing agent, consumes and generates hydroxyl radicals; on the other hand, GSH can degrade Cu... 2+ Reduced to Cu with stronger Fenton-like reactivity + This, in turn, induces the generation of more hydroxyl radicals. This result further corroborates that Cu2(OH)3Cl nanomaterials can, to a certain extent, disrupt the redox dynamic equilibrium dominated by GSH, while simultaneously achieving efficient dual catalytic effects similar to peroxidase and glutathione oxidase.

[0068] (5) Cytotoxicity was assessed using the CCK-8 assay. HeLa (cancer cells, from the Cell Bank of the Chinese Academy of Sciences) and L-929 (normal cells, from the Cell Bank of the Chinese Academy of Sciences) were cultured at 1.5 × 10⁶ cells per well. 4Cells were seeded at a density of [number] cells per well in 96-well plates and cultured at 37 °C and 5% CO2 for 24 hours. Different concentrations (0, 4, 8, 16, 20 μg / mL) of Cu2(OH)3Cl nanomaterials were prepared using cell culture medium. After discarding the old culture medium and washing twice with PBS buffer, the nanomaterials of different concentrations were added to the 96-well plates and incubated for another 24 hours. The old culture medium was then discarded, and the plates were washed twice with PBS buffer. CCK-8 solution was added to each well, and the plates were incubated for another 0.5 hours. The absorbance at 450 nm was measured using a microplate reader.

[0069] Depend on Figure 4 (A) It can be seen that at a concentration of 20 μg / mL, normal cells still have a high survival rate, confirming the good biocompatibility of the nanomaterial. At the same concentration, tumor cells show a low survival rate, indicating that the nanomaterial has a killing effect on tumor cells.

[0070] (6) The Calcein / PI cell staining method was used to further evaluate the killing effect of nanomaterials on HeLa cells. HeLa cells were stained at 1.5 × 10⁶ cells per well. 4 Cells were seeded at a density of [number] cells per well in 96-well plates and cultured at 37 °C and 5% CO2 for 24 hours. A 16 μg / mL concentration of Cu2(OH)3Cl nanomaterial was prepared using culture medium. After discarding the old culture medium and washing twice with PBS buffer, the nanomaterial at the above concentration was added to the 96-well plates and incubated for another 24 hours. The old culture medium was then discarded, and the cells were washed twice with PBS buffer. Cells were stained with Calcein / PI and photographed using a fluorescence inverted microscope. Live cells showed green fluorescence, while dead cells showed red fluorescence.

[0071] Depend on Figure 4 (B) It can be seen that after treatment with 16 μg / mL Cu2(OH)3Cl nanomaterials, tumor cells showed a large number of deaths compared with the untreated group.

[0072] The results of the above cell experiments show that Cu2(OH)3Cl nanomaterials have good biosafety and can amplify peroxidase-like activity by depleting GSH, thereby disrupting the redox homeostasis of tumor cells and achieving a highly efficient tumor killing effect.

[0073] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended 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 therein. Such 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 method for preparing raspberry-like basic copper chloride nanomaterials with dual enzymatic activity, characterized in that, The steps are as follows: Step 1: Add sodium hydroxide aqueous solution to N-fluorenylmethoxycarbonyl-L-arginine (Fmoc-L-Arg), mix by sonication, then add dimethyl sulfoxide to obtain a mixture. Sonicate the mixture until the solution is clear to obtain mixed solution A; prepare copper chloride dihydrate (CuCl2·2H2O) aqueous solution for later use. Step 2: Quickly add CuCl2·2H2O aqueous solution to mixed solution A within one second to obtain mixed solution B, and let it stand to react; Step 3: After the reaction is complete, the precipitate is collected by centrifugation and washed with water to obtain the raspberry-like basic copper chloride nanomaterial with dual enzyme activity, denoted as Cu2(OH)3Cl nanomaterial.

2. The method for preparing a raspberry-like basic copper chloride nanomaterial with dual enzymatic activity according to claim 1, characterized in that, In step one, the ratio of Fmoc-L-Arg, sodium hydroxide aqueous solution, and dimethyl sulfoxide in mixed solution A is 90-110 mg: 200-250 μL: 4-4.5 mL, and the concentration of the added sodium hydroxide aqueous solution is 0.115 M.

3. The method for preparing a raspberry-like basic copper chloride nanomaterial with dual enzymatic activity according to claim 1, characterized in that, In step one, the concentration of the CuCl2·2H2O aqueous solution is 0.16-0.17 M.

4. The method for preparing a raspberry-like basic copper chloride nanomaterial with dual enzymatic activity according to claim 1, characterized in that, In step two, the volume ratio of CuCl2·2H2O aqueous solution to mixed solution A is 1.35-1.5:

1.

5. The method for preparing a raspberry-like basic copper chloride nanomaterial with dual enzymatic activity according to claim 1, characterized in that, In step two, the conditions for the static reaction are: reacting at 24-28 ℃ for 5-8 minutes.

6. The method for preparing a raspberry-like basic copper chloride nanomaterial with dual enzymatic activity according to claim 1, characterized in that, In step three, the centrifugation conditions are: centrifugation at 6000-9000 rpm for 5 minutes.

7. The method for preparing a raspberry-like basic copper chloride nanomaterial with dual enzymatic activity according to claim 1, characterized in that, In step three, the washing with water is as follows: wash with water 1-3 times, and then disperse the washed nanomaterials in ultrapure water for later use.

8. A raspberry-like basic copper chloride nanomaterial with dual enzymatic activity prepared according to any one of claims 1-7.

9. The application of a raspberry-like basic copper chloride nanomaterial with dual enzymatic activity as described in claim 8 in the preparation of a drug for tumor treatment.

Images