Copper-containing lentinan nano-selenium as well as preparation method and application thereof

Abstract

The invention provides copper-containing lentinan nano-selenium as well as application and a preparation method thereof. The preparation method comprises the following steps: (1) preparation of stock solutions: respectively preparing an ascorbic acid solution, a sodium selenite solution, a copper sulfate solution and a lentinan solution; then taking a concentration value within the concentration range according to the ratio of c (CuSO4): c (Na2SeO3) = 1%-100%, and adding water to dilute the copper sulfate solution to obtain a dilute copper sulfate solution; (2) stirring the sodium selenite solution, the dilute copper sulfate solution and the lentinan solution in a reaction bottle, then adding ultrapure water and stirring, then adding an ascorbic acid solution and stirring, then placing the reaction bottle at low temperature for a certain time, and then stirring and reacting at room temperature for a certain time; (3) after the reaction is finished, filling reaction liquid into a dialysis belt, and dialyzing for a certain time; and after dialysis is finished, transferring the reaction liquid, centrifuging, and taking the supernatant to obtain the target product copper-containing lentinan nano-selenium. The invention aims to explore the application of the copper-containing nano-selenium medicine in the aspect of tumor treatment.

Description

Technical Field

[0001] The invention belongs to the technical field of synthetic drug chemistry, and particularly relates to a copper-containing lentinan nano-selenium, a preparation method of the copper-containing lentinan nano-selenium, and an application of the copper-containing lentinan nano-selenium in drugs for treating malignant tumors. Background Art

[0002] Cancer is one of the biggest threats to human health. The latest global cancer data in 2022 shows that there will be nearly 20 million new cancer cases in the world in 2022, among which the incidence and mortality of lung cancer and breast cancer are at the forefront. At present, the main means of cancer treatment include chemotherapy, radiotherapy, surgery, immunotherapy, targeted therapy and combined therapy. Chemotherapy is one of the commonly used means of cancer treatment. Although chemotherapy drugs are rich and diverse, the side effects they bring, such as drug resistance and systemic toxicity, are still a major obstacle to this therapy.

[0003] Selenium (Se), as one of the essential trace elements for human health, plays a vital role in preventing diseases. By binding to selenoproteins, selenium exerts multiple effects, including antioxidant, anti-inflammatory, anti-tumor and immunomodulatory. Chemical derivatives of selenium include inorganic compounds such as selenite and organic compounds such as selenomethylselenocysteine ​​and selenomethionine. In recent years, more and more studies have demonstrated that selenium has a chemopreventive effect in cancer risk and incidence, but the inorganic and organic forms of selenium are still highly toxic and prone to side effects. After nano-sizing selenium, selenium can have higher biocompatibility and utilization, and more importantly, it can reduce toxicity while maintaining high biological activity.

[0004] Transition metals, due to their unique d or f orbitals, have highly efficient catalytic activity, inducing the production of reactive oxygen species (ROS) through the Fenton reaction, thereby increasing cellular oxidative stress, leading to DNA, protein and lipid damage, and ultimately causing cancer cell death. Transition metals represented by copper have been widely studied as components of metalloenzymes, signaling molecules and important components of metal anticancer drugs. In addition, copper-based drugs also have the advantages of relatively low cost, adjustable structure, diverse composition, efficient photothermal / photodynamic therapy performance, unique imaging capabilities and good biocompatibility. Based on this, in order to improve the anti-tumor activity of nano-selenium, it is of great significance to design nano-selenium and copper doping to form selenium-copper compounds with other functions such as catalytic activity, and to study their anti-tumor mechanisms. Summary of the invention

[0005] In view of the above-mentioned shortcomings of the prior art, the present invention aims to explore the application of copper-containing nano-selenium drugs obtained by doping copper sulfate and lentinan nano-selenium in different proportions in the treatment of tumors.

[0006] The present application provides a method for preparing copper-containing Lentinan nano-selenium, comprising the following steps:

[0007] (1) Preparation of stock solutions: ascorbic acid solution, sodium selenite solution, copper sulfate solution and lentinan solution are prepared respectively; then, the copper sulfate solution is diluted with water at a concentration value within the concentration range of c(CuSO4):c(Na2SeO3)=1%-100% to obtain a dilute copper sulfate solution; it should be noted that the copper sulfate solution is diluted with water at a ratio of c(CuSO4):c(Na2SeO3) refers to the concentration ratio of the diluted copper sulfate solution to the sodium selenite solution.

[0008] (2) stirring a sodium selenite solution, a dilute copper sulfate solution and a lentinan solution in a reaction flask, then adding ultrapure water and stirring, then adding an ascorbic acid solution and stirring, then placing the reaction flask at low temperature for a certain period of time, and then stirring and reacting at room temperature for a certain period of time;

[0009] (3) After the reaction is completed, the reaction solution is placed in a dialysis tape and dialyzed for a certain period of time; after the dialysis is completed, the reaction solution is transferred and centrifuged, and the supernatant is taken to obtain the target product, copper-containing Lentinan polysaccharide nano-selenium.

[0010] Specifically, the concentration ranges of the ascorbic acid solution, sodium selenite solution, copper sulfate solution and lentinan solution are 180-220 mM, 40-60 mM, 40-60 mM and 11.5-13.5 mg / mL, respectively.

[0011] Specifically, c(CuSO4):c(Na2SeO3)=any one of 1%, 10%, 30%, and 100%.

[0012] Specifically, step (2): stir the sodium selenite solution, dilute copper sulfate solution and lentinan solution in a reaction bottle for 4-6 minutes, then add ultrapure water and stir for 4-6 minutes, then add ascorbic acid solution and stir for 4-6 minutes, then place the reaction bottle at 3-5°C for 25-35 minutes, and then stir and react at room temperature for 11-13 hours.

[0013] Specifically, step (3): after the reaction is completed, the reaction solution is loaded into a 6000-8000 molecular weight dialysis tape and dialyzed for 2.5-3.5 days; after the dialysis is completed, the reaction solution is transferred and centrifuged at 2500-3500 rpm for 9-11 minutes, and the supernatant is taken to obtain the target product, copper-containing Lentinan polysaccharide nano-selenium.

[0014] Specifically, the following steps are included:

[0015] (1) Preparation of stock solution: Weigh a certain amount of ascorbic acid powder and dissolve it in ultrapure water to prepare a 200 mM ascorbic acid solution; weigh a certain amount of sodium selenite powder and dissolve it in ultrapure water to prepare a 50 mM sodium selenite solution; weigh a certain amount of copper sulfate pentahydrate and dissolve it in ultrapure water to prepare a 50 mM copper sulfate solution, and dilute the copper sulfate solution with water at any concentration value within the concentration range of c(CuSO4):c(Na2SeO3)=10%, 30%, 100% to obtain a dilute copper sulfate solution; weigh 125 mg of Lentinan powder in a centrifuge tube, add ultrapure water to a total volume of 10 mL, and prepare a 12.5 mg / mL Lentinan solution;

[0016] (2) 1 mL of sodium selenite solution, 1 mL of dilute copper sulfate solution and 2 mL of Lentinan solution were added to the reaction bottle and stirred for 5 min, then 5 mL of ultrapure water was added and stirred for 5 min, then 1 mL of ascorbic acid solution was added and stirred for 5 min, then the reaction bottle was placed at 4 ° C for 30 min, and then stirred at room temperature for 12 h;

[0017] (3) After the reaction is completed, the reaction solution is loaded into a 6000-8000 molecular weight dialysis tape and dialyzed for 3 days. After the dialysis is completed, the reaction solution is transferred and centrifuged at 3000 rpm for 10 min, and the supernatant is taken to obtain the target product, copper-containing Lentinan nano-selenium.

[0018] Copper-containing lentinan nano-selenium prepared according to any of the preparation methods described above.

[0019] The application of the copper-containing lentinan nano-selenium in drugs for treating malignant tumors.

[0020] The improvements of the present application bring the following advantages: The present invention has been experimentally proven that the introduction of copper can significantly enhance the inhibitory effect of Lentinan nano-selenium on tumor growth, making these copper-containing nano-selenium compounds promising for the treatment of tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a hydrated particle size diagram of copper-containing Lentinan nano-selenium in the embodiment of the present application;

[0022] Figure 2 This is the XRD diagram of copper-containing Lentinan nano-selenium in the embodiment of this application;

[0023] Figure 3 This is a TEM image of copper-containing Lentinan nano-selenium in the example of this application. DETAILED DESCRIPTION

[0024] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0025] Example 1: Preparation of copper-containing Lentinan nano-selenium with different doping ratios

[0026] (1) Preparation of stock solution: Weigh a certain amount of ascorbic acid powder and dissolve it in ultrapure water to prepare a 200 mM ascorbic acid solution; weigh a certain amount of sodium selenite powder and dissolve it in ultrapure water to prepare a 50 mM sodium selenite solution; weigh a certain amount of copper sulfate pentahydrate and dissolve it in ultrapure water to prepare a 50 mM copper sulfate solution, dilute the copper sulfate solution according to the concentration of c(CuSO4):c(Na2SeO3)=10%, and obtain a dilute copper sulfate solution; weigh 125 mg of Lentinan powder in a centrifuge tube, add ultrapure water to a total volume of 10 mL, and prepare a 12.5 mg / mL Lentinan solution;

[0027] (2) 1 mL of sodium selenite solution, 1 mL of dilute copper sulfate solution and 2 mL of Lentinan solution were added to the reaction bottle and stirred for 5 min, then 5 mL of ultrapure water was added and stirred for 5 min, then 1 mL of ascorbic acid solution was added and stirred for 5 min, then the reaction bottle was placed at 4 ° C for 30 min, and then stirred at room temperature for 12 h;

[0028] (3) After the reaction is completed, the reaction solution is loaded into a 7000 molecular weight dialysis tape and dialyzed for 3 days. After the dialysis is completed, the reaction solution is transferred and centrifuged at 3000 rpm for 10 min, and the supernatant is taken to obtain the target product, copper-containing Lentinan nano-selenium.

[0029] (4) Repeat steps (1)-(3) twice to prepare multiple copper-containing Lentinan nano-selenium with different doping ratios, the difference being that the dilution ratio of the copper sulfate solution in step (1) is different, and the copper sulfate solutions of different concentrations are obtained by diluting the solution according to three different concentrations of c(CuSO4):c(Na2SeO3)=1%, 30% and 100%, respectively, to prepare copper-containing Lentinan nano-selenium with different doping ratios.

[0030] Example 2: Preparation of copper-containing Lentinan nano-selenium with different doping ratios

[0031] (1) Preparation of stock solution: Weigh a certain amount of ascorbic acid powder and dissolve it in ultrapure water to prepare a 180 mM ascorbic acid solution; weigh a certain amount of sodium selenite powder and dissolve it in ultrapure water to prepare a 40 mM sodium selenite solution; weigh a certain amount of copper sulfate pentahydrate and dissolve it in ultrapure water to prepare a 40 mM copper sulfate solution, dilute the copper sulfate solution according to the concentration of c(CuSO4):c(Na2SeO3)=10%, and obtain a dilute copper sulfate solution; weigh 115 mg of Lentinan powder in a centrifuge tube, add ultrapure water to a total volume of 10 mL, and prepare a 11.5 mg / mL Lentinan solution;

[0032] (2) 0.9 mL of sodium selenite solution, 0.9 mL of dilute copper sulfate solution and 1.8 mL of Lentinan solution were added to the reaction bottle and stirred for 4 min, then 4 mL of ultrapure water was added and stirred for 4 min, then 0.9 mL of ascorbic acid solution was added and stirred for 4 min, then the reaction bottle was placed at 5 ° C for 25 min, and then stirred at room temperature for 11 h;

[0033] (3) After the reaction is completed, the reaction solution is loaded into a 6000 molecular weight dialysis tape and dialyzed for 2.5 days. After the dialysis is completed, the reaction solution is transferred and centrifuged at 2500 rpm for 9 minutes, and the supernatant is taken to obtain the target product, copper-containing Lentinan nano-selenium.

[0034] (4) Repeat steps (1)-(3) twice to prepare multiple copper-containing Lentinan nano-selenium with different doping ratios, the difference being that the dilution ratio of the copper sulfate solution in step (1) is different, and the copper sulfate solutions of different concentrations are obtained by diluting the solution according to three different concentrations of c(CuSO4):c(Na2SeO3)=1%, 30% and 100%, respectively, to prepare copper-containing Lentinan nano-selenium with different doping ratios.

[0035] Example 3: Preparation of copper-containing Lentinan nano-selenium with different doping ratios

[0036] (1) Preparation of stock solution: Weigh a certain amount of ascorbic acid powder and dissolve it in ultrapure water to prepare a 220 mM ascorbic acid solution; weigh a certain amount of sodium selenite powder and dissolve it in ultrapure water to prepare a 60 mM sodium selenite solution; weigh a certain amount of copper sulfate pentahydrate and dissolve it in ultrapure water to prepare a 60 mM copper sulfate solution, dilute the copper sulfate solution according to the concentration of c(CuSO4):c(Na2SeO3)=10%, and obtain a dilute copper sulfate solution; weigh 135 mg of Lentinan powder in a centrifuge tube, add ultrapure water to a total volume of 10 mL, and prepare a 13.5 mg / mL Lentinan solution;

[0037] (2) 1.1 mL of sodium selenite solution, 1.1 mL of dilute copper sulfate solution and 2.2 mL of Lentinan solution were added to the reaction bottle and stirred for 6 min, then 6 mL of ultrapure water was added and stirred for 6 min, then 1.1 mL of ascorbic acid solution was added and stirred for 6 min, then the reaction bottle was placed at 3 ° C for 35 min, and then stirred at room temperature for 13 h;

[0038] (3) After the reaction is completed, the reaction solution is loaded into a 8000 molecular weight dialysis tape and dialyzed for 3.5 days. After the dialysis is completed, the reaction solution is transferred and centrifuged at 3500 rpm for 11 minutes, and the supernatant is taken to obtain the target product, copper-containing Lentinan nano-selenium.

[0039] (4) Repeat steps (1)-(3) twice to prepare multiple copper-containing Lentinan nano-selenium with different doping ratios, the difference being that the dilution ratio of the copper sulfate solution in step (1) is different, and the copper sulfate solutions of different concentrations are obtained by diluting the solution according to three different concentrations of c(CuSO4):c(Na2SeO3)=1%, 30% and 100%, respectively, to prepare copper-containing Lentinan nano-selenium with different doping ratios.

[0040] Example 4: Preparation of copper-containing Lentinan nano-selenium with different doping ratios

[0041] (1) Preparation of stock solution: Weigh a certain amount of ascorbic acid powder and dissolve it in ultrapure water to prepare a 190 mM ascorbic acid solution; weigh a certain amount of sodium selenite powder and dissolve it in ultrapure water to prepare a 45 mM sodium selenite solution; weigh a certain amount of copper sulfate pentahydrate and dissolve it in ultrapure water to prepare a 45 mM copper sulfate solution, dilute the copper sulfate solution according to the concentration of c(CuSO4):c(Na2SeO3)=10%, and obtain a dilute copper sulfate solution; weigh 120 mg of Lentinan powder in a centrifuge tube, add ultrapure water to a total volume of 10 mL, and prepare a 12 mg / mL Lentinan solution;

[0042] (2) 1 mL of sodium selenite solution, 1 mL of dilute copper sulfate solution and 2 mL of Lentinan solution were added to the reaction bottle and stirred for 5 min, then 5 mL of ultrapure water was added and stirred for 5 min, then 1 mL of ascorbic acid solution was added and stirred for 5 min, then the reaction bottle was placed at 4 ° C for 25 min, and then stirred at room temperature for 11 h;

[0043] (3) After the reaction is completed, the reaction solution is loaded into a 6500 molecular weight dialysis tape and dialyzed for 3 days. After the dialysis is completed, the reaction solution is transferred and centrifuged at 2800 rpm for 10 min, and the supernatant is taken to obtain the target product, copper-containing Lentinan nano-selenium.

[0044] (4) Repeat steps (1)-(3) twice to prepare multiple copper-containing Lentinan nano-selenium with different doping ratios, the difference being that the dilution ratio of the copper sulfate solution in step (1) is different, and the copper sulfate solutions of different concentrations are obtained by diluting the solution according to three different concentrations of c(CuSO4):c(Na2SeO3)=1%, 30% and 100%, respectively, to prepare copper-containing Lentinan nano-selenium with different doping ratios.

[0045] Example 5: Preparation of copper-containing Lentinan nano-selenium with different doping ratios

[0046] (1) Preparation of stock solution: Weigh a certain amount of ascorbic acid powder and dissolve it in ultrapure water to prepare a 210 mM ascorbic acid solution; weigh a certain amount of sodium selenite powder and dissolve it in ultrapure water to prepare a 55 mM sodium selenite solution; weigh a certain amount of copper sulfate pentahydrate and dissolve it in ultrapure water to prepare a 55 mM copper sulfate solution, dilute the copper sulfate solution according to the concentration of c(CuSO4):c(Na2SeO3)=10%, and obtain a dilute copper sulfate solution; weigh 125 mg of Lentinan powder in a centrifuge tube, add ultrapure water to a total volume of 10 mL, and prepare a 12.5 mg / mL Lentinan solution;

[0047] (2) 1 mL of sodium selenite solution, 1 mL of dilute copper sulfate solution and 2 mL of Lentinan solution were added to the reaction bottle and stirred for 5 min, then 5 mL of ultrapure water was added and stirred for 5 min, then 1 mL of ascorbic acid solution was added and stirred for 5 min, then the reaction bottle was placed at 4 ° C for 30 min, and then stirred at room temperature for 12 h;

[0048] (3) After the reaction is completed, the reaction solution is loaded into a 7500 molecular weight dialysis tape and dialyzed for 3 days. After the dialysis is completed, the reaction solution is transferred and centrifuged at 3300 rpm for 10 min, and the supernatant is taken to obtain the target product, copper-containing Lentinan nano-selenium.

[0049] (4) Repeat steps (1)-(3) twice to prepare multiple copper-containing Lentinan nano-selenium with different doping ratios, the difference being that the dilution ratio of the copper sulfate solution in step (1) is different, and the copper sulfate solutions of different concentrations are obtained by diluting the solution according to three different concentrations of c(CuSO4):c(Na2SeO3)=1%, 30% and 100%, respectively, to prepare copper-containing Lentinan nano-selenium with different doping ratios.

[0050] Experiment 6: Characterization of Copper-Containing Lentinan Nanoselenium

[0051] (1) ICP: 100 μL of sample (containing copper-containing Lentinan nano-selenium) was nitrated with aqua regia and diluted to 10 mL with ultrapure water. The sample was tested on an iCAP-RQ inductively coupled plasma mass spectrometer. As shown in Table 1, copper can be doped into Lentinan nano-selenium at a theoretical doping ratio.

[0052] Table 1 Theoretical and actual doping of copper-containing Lentinan nano-selenium drugs

[0053]

[0054] (2) Particle size: Take 200 μL of sample and dilute it to 2 mL with ultrapure water. Perform particle size test on Malvern potentiometric particle size analyzer. Figure 1 As shown, it indicates that the copper-containing Lentinan nano-selenium has a suitable particle size.

[0055] (3) XRD: Take a small amount of sample powder and test it on an X-ray diffractometer, such as Figure 2 As shown, the incorporation of Cu ions causes the XRD peak to shift to the right.

[0056] (4) TEM: Take 200 μL of sample and drop it on the copper grid, then test it under transmission electron microscope, such as Figure 3 As shown, the particle size decreases with the increase of copper doping ratio.

[0057] Experiment 7: Study on the anti-tumor activity of copper-containing Lentinan nanoselenium in vitro

[0058] The method of measuring the growth inhibition of tumor cells and normal cells (HeLa, HepG2, HK2 cells) by using the MTT method in Experiment 1 mainly includes the following steps:

[0059] (1) Place the copper-containing Lentinan nanoselenium on an ultrasonicator for 2 minutes, take a certain amount of sample and dilute it to 64 μg / mL with DMEM medium, and perform gradient dilution, and the solution is ready for use;

[0060] (2) Different tumor cells in logarithmic growth phase (American Model Culture Collection) were obtained at a density of 2×10 3cells / mL were inoculated into a 96-well plate (100 μL / well) and allowed to adhere to the wall and grow for 24 hours. Then, 100 μL of the drugs of different concentrations in step (1) were added respectively and cultured for 72 hours. After the cells showed a certain degree of damage, 30 μL of MTT solution (5 mg / mL, PBS solution) was added to each well and incubated for 4 hours. Then, the upper culture medium (DMEM high-glucose medium, Gibco) in the 96-well plate was aspirated, 150 μL of dimethyl sulfoxide was added, and the plate was gently shaken on a shaker for 15 minutes to fully dissolve the purple crystals in the 96-well plate. Then, the absorbance of each well at 570 nm was measured with a multifunctional microplate reader, and the cell survival rate was calculated, and a graph was drawn to obtain the half inhibitory concentration (IC50). Cell survival rate (%) = (OD570 experimental group / OD570 control group) × 100%.

[0061] Test results: As shown in Table 2, the introduction of selenium can significantly enhance the inhibitory effect of copper-containing Lentinan nano-selenium drugs on tumor growth. Compared with undoped Lentinan nano-selenium, copper-containing Lentinan nano-selenium has a stronger inhibitory effect on tumor cells, and the inhibition on HeLa cells is more obvious.

[0062] Table 2 Results of MTT toxicity test on tumor cells of copper-containing Lentinan nanoselenium drugs

[0063]

[0064] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A method for preparing copper-containing Lentinan nano-selenium, characterized in that: The following steps are involved: (1) Preparation of stock solutions: preparing ascorbic acid solution, sodium selenite solution, copper sulfate solution and lentinan solution respectively; then diluting the copper sulfate solution with water at a concentration value within the concentration range of c(CuSO4):c(Na2SeO3)=1%-100% to obtain a dilute copper sulfate solution; (2) stirring a sodium selenite solution, a dilute copper sulfate solution and a lentinan solution in a reaction flask, then adding ultrapure water and stirring, then adding an ascorbic acid solution and stirring, then placing the reaction flask at low temperature for a certain period of time, and then stirring and reacting at room temperature for a certain period of time; (3) After the reaction is completed, the reaction solution is placed in a dialysis tape and dialyzed for a certain period of time; after the dialysis is completed, the reaction solution is transferred and centrifuged, and the supernatant is taken to obtain the target product, copper-containing Lentinan polysaccharide nano-selenium.

2. The preparation method according to claim 1, characterized in that: The concentration ranges of the ascorbic acid solution, the sodium selenite solution, the copper sulfate solution and the lentinan solution are 180-220 mM, 40-60 mM, 40-60 mM and 11.5-13.5 mg / mL respectively.

3. The preparation method according to claim 1, characterized in that: c(CuSO4):c(Na2SeO3)=1%, 10%, 30%, 100% any one.

4. The preparation method according to claim 1, characterized in that: Step (2): Stir the sodium selenite solution, dilute copper sulfate solution and Lentinan solution in a reaction flask for 4-6 minutes, then add ultrapure water and stir for 4-6 minutes, then add ascorbic acid solution and stir for 4-6 minutes, then place the reaction flask at 3-5°C for 25-35 minutes, and then stir and react at room temperature for 11-13 hours.

5. The preparation method according to claim 1, characterized in that: Step (3): After the reaction is completed, the reaction solution is loaded into a 6000-8000 molecular weight dialysis tape and dialyzed for 2.5-3.5 days; after the dialysis is completed, the reaction solution is transferred and centrifuged at 2500-3500 rpm for 9-11 minutes, and the supernatant is taken to obtain the target product, copper-containing Lentinan polysaccharide nano-selenium.

6. The preparation method according to any one of claims 1 to 5, characterized in that: The following steps are involved: (1) Preparation of stock solution: Weigh a certain amount of ascorbic acid powder and dissolve it in ultrapure water to prepare a 200 mM ascorbic acid solution; weigh a certain amount of sodium selenite powder and dissolve it in ultrapure water to prepare a 50 mM sodium selenite solution; weigh a certain amount of copper sulfate pentahydrate and dissolve it in ultrapure water to prepare a 50 mM copper sulfate solution, and dilute the copper sulfate solution with water at any concentration value within the concentration range of c(CuSO4):c(Na2SeO3)=10%, 30%, 100% to obtain a dilute copper sulfate solution; weigh 125 mg of Lentinan powder in a centrifuge tube, add ultrapure water to a total volume of 10 mL, and prepare a 12.5 mg / mL Lentinan solution; (2) 1 mL of sodium selenite solution, 1 mL of dilute copper sulfate solution and 2 mL of Lentinan solution were added to the reaction bottle and stirred for 5 min, then 5 mL of ultrapure water was added and stirred for 5 min, then 1 mL of ascorbic acid solution was added and stirred for 5 min, then the reaction bottle was placed at 4 ° C for 30 min, and then stirred at room temperature for 12 h; (3) After the reaction is completed, the reaction solution is loaded into a 6000-8000 molecular weight dialysis tape and dialyzed for 3 days. After the dialysis is completed, the reaction solution is transferred and centrifuged at 3000 rpm for 10 min, and the supernatant is taken to obtain the target product, copper-containing Lentinan nano-selenium.

7. Copper-containing Lentinan nano-selenium obtained according to the preparation method according to any one of claims 1 to 6.

8. Use of the copper-containing Lentinan nano-selenium according to claim 7 in drugs for treating malignant tumors.

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