Medicine for treating hepatocellular carcinoma and application

Abstract

The invention discloses a medicine for treating hepatocellular carcinoma and application. The preparation method comprises the following steps: firstly, constructing selenium-based nanoparticles Se / ZIF8, wherein the selenium-based nanoparticles consist of Se and ZIF8; the selenium-based nanoparticles and the NK cells are jointly used for treating the liver cancer cells, so that the killing activity of the NK cells on the liver cancer cells can be remarkably enhanced. Meanwhile, the selenium-based nanoparticles and the NK cells are used for jointly treating in-situ liver cancer tumor-bearing mice, the tumor inhibition effect of the NK cells can be remarkably improved, and the synergistic interaction effect is achieved. The invention provides a new immune drug choice for treating the liver cancer, and has a wide application prospect in clinical treatment of the liver cancer.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and specifically, to a drug for treating hepatocellular carcinoma and its application. Background Art

[0002] Hepatocellular carcinoma (HCC) is the most common primary liver cancer globally. Its etiology is complex and diverse, and the mortality rate is high, making it the fourth leading cause of cancer-related deaths. With the progress of medical research, treatment strategies for HCC are constantly evolving, especially in the fields of targeted therapy and immunotherapy. Traditional chemotherapy has limited efficacy against HCC, while emerging targeted therapy and immunotherapy offer more hope.

[0003] The immune microenvironment of the liver is rich in macrophages, NK cells, and T lymphocytes, providing a unique opportunity for the development of effective HCC immunotherapies. Notably, NK cells account for 25% - 50% of liver lymphocytes, a proportion significantly higher than that in the spleen or peripheral blood, indicating their crucial role in liver immune defense. A key advantage of NK cells is their ability to mediate anti-tumor effects independent of major histocompatibility complex (MHC) restriction, enabling them to rapidly and directly eliminate cancer cells. NK cells exert their anti-tumor functions through two main mechanisms: direct cytotoxicity through receptor-mediated tumor cell recognition and induction of apoptosis via death receptors, and indirect cytotoxicity through the secretion of perforin, granzyme B, and pro-inflammatory cytokines.

[0004] Although NK cell immunotherapy is effective in hematological malignancies, it has not produced ideal results in solid tumors. As a deep-seated malignancy, HCC has an immunosuppressive tumor microenvironment (TME) that can impede NK cell infiltration, inhibit immune activation, and lead to NK cell dysfunction. Elevated levels of reactive oxygen species (ROS) within the tumor further limit NK cell-mediated tumor eradication by causing NK cell damage and dysfunction. Therefore, the development of a drug that effectively promotes NK cell therapy for HCC is of great significance.

[0005] Selenium (Se) is an essential trace element that is metabolized in the body to selenocysteine (Sec) residues and incorporated into 25 selenoproteins that regulate antioxidant defense and energy metabolism. However, the biomedical application of elemental selenium is limited by its poor water solubility, so strategies are needed to improve its bioavailability. Summary of the Invention

[0006] To overcome the above-mentioned defects and deficiencies in the prior art, the present invention provides a drug for treating hepatocellular carcinoma and its application.

[0007] The first object of the present invention is to provide an application of a composition in the preparation of a medicament for treating liver cancer.

[0008] The second object of the present invention is to provide a medicament for treating liver cancer.

[0009] The present invention claims the following: An application of a composition in the preparation of a medicament for treating liver cancer, wherein the composition comprises selenium-based nanoparticles Se / ZIF8 and NK cells; The preparation method of the selenium-based nanoparticles Se / ZIF8 comprises the following steps: fully reacting selenium powder with zeolitic imidazolate framework material ZIF8 to obtain the product.

[0010] Preferably, the preparation method of the selenium-based nanoparticles Se / ZIF8 comprises the following steps: fully reacting selenium powder with zeolitic imidazolate framework material ZIF8, and then modifying the product with DSPE-PEG2000 to obtain the product.

[0011] Preferably, the preparation method of the zeolitic imidazolate framework material ZIF8 is: mixing zinc salt, 2-methylimidazole and methanol and fully reacting them to obtain the product.

[0012] More preferably, the dosage ratio of the zinc salt, 2-methylimidazole and methanol is (1-1.4) g: (2.4-2.8) g: (78-82) mL.

[0013] Further preferably, the dosage ratio of the zinc salt, 2-methylimidazole and methanol is 1.2 g: 2.6 g: 80 mL.

[0014] More preferably, the reaction temperature is 24-26 °C and the reaction time is 0.8-1.2 h.

[0015] Further preferably, the reaction time is 1 h.

[0016] Further preferably, the zinc salt is Zn(NO3)2·6H2O.

[0017] Preferably, the dosage ratio of the selenium powder and the zeolitic imidazolate framework material ZIF8 is 1 g: (3-5) g.

[0018] More preferably, the dosage ratio of the selenium powder and the zeolitic imidazolate framework material ZIF8 is 1 g: 4 g.

[0019] Preferably, the temperature of the full reaction is 255-265 °C.

[0020] More preferably, the temperature of the full reaction is 260 °C.

[0021] Preferably, the time for sufficient reaction is 0.8 to 1.2 h.

[0022] More preferably, the time for sufficient reaction is 1 h.

[0023] Preferably, the dosage ratio of the product to DSPE-PEG2000 is 1 g:(0.8 - 1.2) g.

[0024] More preferably, the dosage ratio of the product to DSPE-PEG2000 is 1 g:1 g.

[0025] Preferably, the modification is to react the product with DSPE-PEG2000 for 11 to 13 h after mixing.

[0026] More preferably, the modification is to react the product with DSPE-PEG2000 for 12 h after mixing.

[0027] Preferably, the dosage ratio of the selenium-based nanoparticle Se / ZIF8 to NK cells in the composition is (0.125 - 1) μg:(0.8 - 1.2)×10 7 cells.

[0028] More preferably, the dosage ratio of the selenium-based nanoparticle Se / ZIF8 to NK cells is 1 μg:1×10 7 cells.

[0029] Preferably, the liver cancer is primary liver cancer.

[0030] More preferably, the primary liver cancer is hepatocellular carcinoma.

[0031] An anti-tumor drug, wherein the anti-tumor drug comprises the above composition.

[0032] Preferably, the dosage ratio of the selenium-based nanoparticle Se / ZIF8 to NK cells in the composition is (0.125 - 1) μg:(0.8 - 1.2)×10 7 cells.

[0033] More preferably, the dosage ratio of the selenium-based nanoparticle Se / ZIF8 to NK cells is 1 μg:1×10 7 cells.

[0034] Preferably, the liver cancer is primary liver cancer.

[0035] More preferably, the primary liver cancer is hepatocellular carcinoma.

[0036] Compared with the prior art, the present invention has the following beneficial effects: The present invention discloses a drug for treating hepatocellular carcinoma and its application. First, the selenium-based nanoparticles Se / ZIF8 are constructed in the present invention, and the selenium-based nanoparticles are composed of Se and zeolitic imidazolate framework material ZIF8. When the selenium-based nanoparticles are co-treated with NK cells on liver cancer cells, the killing activity of NK cells against liver cancer cells can be significantly enhanced. At the same time, when the selenium-based nanoparticles are co-treated with NK cells on mice bearing orthotopic liver cancer tumors, the tumor suppression effect of NK cells can be significantly improved, playing a role of synergistic enhancement. The preparation raw materials of the selenium-based nanoparticles of the present invention are cheap and easily available, and the preparation method is simple and efficient, providing a new choice of immune drug for the treatment of liver cancer and having broad application prospects in the clinical treatment of liver cancer. Description of the Drawings

[0037] Figure 1 Zeta potential characterization results of ZIF8, intermediate and Se / ZIF8 prepared in Example 1.

[0038] Figure 2 Transmission electron microscope morphology characterization diagrams of ZIF8, intermediate and Se / ZIF8 prepared in Example 1.

[0039] Figure 3 Element mapping analysis diagrams of ZIF8, intermediate and Se / ZIF8 prepared in Example 1.

[0040] Figure 4 Survival rate results of NK cells killing HepG-2 cells at different effector-to-target ratios.

[0041] Figure 5 Effects of ZIF8, intermediate and Se / ZIF8 at different concentrations on the survival rate of liver cancer cells HepG-2.

[0042] Figure 6 Effects of different treatment methods of Se / ZIF8 and NK cells on the survival rate of HepG-2 cells.

[0043] Figure 7 Changes in Se content in plasma over time after intravenous injection of the intermediate and Se / ZIF8 (n = 5).

[0044] Figure 8 Effects of different materials acting alone or in combination with NK cells on the survival rate of HepG-2 cells.

[0045] Figure 9 Killing effects of Se / ZIF8, NK cells or Se / ZIF8+NK cells on in vitro tumor spheres.

[0046] Figure 10Animal magnetic resonance imaging of the effect of Se / ZIF8, NK cells, or Se / ZIF8 + NK cells on tumor development in an orthotopic liver cancer-bearing mouse model established with HepG-2 tumor cells.

[0047] Figure 11 Comparison chart of the differences in the development of tumors and the liver in an orthotopic liver cancer-bearing mouse model established with HepG-2 tumor cells treated with Se / ZIF8, NK cells, or Se / ZIF8 + NK cells.

[0048] Figure 12 Influence diagram of the change in tumor growth volume in an orthotopic liver cancer-bearing mouse model established with HepG-2 tumor cells treated with Se / ZIF8, NK cells, or Se / ZIF8 + NK cells.

[0049] Figure 13 Influence diagram of the change in tumor growth weight in an orthotopic liver cancer-bearing mouse model established with HepG-2 tumor cells treated with Se / ZIF8, NK cells, or Se / ZIF8 + NK cells.

[0050] Figure 14 Comparison chart of the tumor growth inhibition rate in an orthotopic liver cancer-bearing mouse model established with HepG-2 tumor cells treated with Se / ZIF8, NK cells, or Se / ZIF8 + NK cells. Detailed implementation mode

[0051] The following specific examples are used to further illustrate the present invention, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in the technical field.

[0052] Unless otherwise specified, the reagents and materials used in the following examples are all commercially available.

[0053] Example 1 Preparation and characterization of Se / ZIF8 I. Experimental method Dissolve 1.2 g of Zn(NO3)2·6H2O and 2.6 g of 2-methylimidazole in 80 mL of methanol, stir and react at room temperature for 1 h to obtain the zeolitic imidazolate framework material ZIF8. After drying ZIF8, mix it with Se powder in a mass ratio of 4 g:1 g in a vacuum-sealed environment, and then heat it to 260 °C and keep it warm for 1 h to obtain an intermediate. Mix 100 mg of the intermediate with 10 mL of an aqueous solution of DSPE-PEG2000 with a concentration of 10 g / L, stir for 12 h, then centrifuge at 10000 rpm for 10 min to take the precipitate, and freeze-dry the precipitate to obtain Se / ZIF8.

[0054] Zeta potential and transmission electron microscopy were used to characterize ZIF8, intermediates and Se / ZIF8 using Zetasizer Nano ZS, and the chemical compositions of ZIF8, intermediates and Se / ZIF8 were determined using Fourier transform infrared spectrometer (Equinox 55, Bruker).

[0055] 2. Experimental Results Zeta potential characterization results are shown in Figure 1 As shown in the figure, the surface of ZIF8 is mostly positively charged, and the loading of Se (i.e., intermediate) changes the Zeta potential from positive to negative, confirming the successful incorporation of Se. After DSPE-PEG2000 modification (i.e., Se / ZIF8), the Zeta potential is slightly negative and has no significant change.

[0056] The morphological characterization results of transmission electron microscopy (TEM) are shown in Figure 2 As shown, ZIF8 alone exhibited a regular morphology and good dispersion, but Se-loaded (i.e., intermediate) and DSPE-PEG2000-modified nanomaterials (i.e., Se / ZIF8) presented a more rounded shape.

[0057] Fourier transform infrared spectrometer test results ( Figure 3 ) further confirmed the successful synthesis of the nanomaterial (i.e., Se / ZIF8), and in addition to the characteristic N and Zn signals from ZIF8, the uniform distribution of Se throughout the MOF and the presence of P element from DSPE-PEG2000 could be clearly detected.

[0058] Example 2 Optimal target ratio of NK cells to kill HepG-2 cells 1. Experimental Methods 1. Cultivation of HepG-2 cells and NK-92 cells HepG-2 cells were purchased from the American Type Culture Collection (ATCC) and added with DMEM complete medium containing 10% fetal bovine serum (v / v) and 1% double antibody (v / v), and cultured statically in a humidified incubator at 37°C and 5% CO2.

[0059] NK-92 cells were purchased from the American Type Culture Collection (ATCC) and added with α-MEM complete medium containing 10% fetal bovine serum (v / v), 10% horse serum (v / v) and 1% double antibody (v / v), and cultured statically in a humidified incubator at 37°C and 5% CO2.

[0060] 2. MTT assay Plate HepG-2 cells at a density of 5000 cells per well. After culturing HepG-2 cells for 24 h, add NK-92 cells (plate NK-92 cells at a cell number ratio of NK-92:HepG-2 of 1:4, 1:2, 1:1, 2:1, and 4:1) and incubate for 72 h. After the treatment, gently aspirate the upper layer of NK-92 cells, add 100 μL of PBS buffer to each well, and take pictures of HepG-2 cells with a Nikon microscope.

[0061] Use an MTT assay kit to detect the survival rate of HepG-2 cells. The specific steps are as follows: Add 25 μL of MTT solution (5 mg / mL, Sigma) to each well and incubate in a 37°C, 5% CO2 incubator for 4 - 5 h. After aspirating the supernatant, add 150 μL of DMSO to each well. Use the group without adding NK-92 cells as the control group, read the value at 570 nm with an enzyme-linked immunosorbent assay (ELISA) reader, and calculate the survival rate of the cells. The survival rate of the cells is calculated according to the following formula: Survival rate = absorbance value of the treatment group / absorbance value of the control group × 100%.

[0062] II. Experimental Results The results are as Figure 4 shown. The cytotoxicity of NK cells against HepG-2 cells increases with the increase in the effector-to-target ratio (the cell number ratio of NK cells to HepG-2 cells). Therefore, a 1:1 effector-to-target ratio was selected for the tumor killing experiment.

[0063] Example 3 Effect of Se / ZIF8 on the Survival Rate of HepG-2 Cells I. Experimental Method Seed HepG-2 cells in a cell culture plate, add 100 μL of DMEM complete medium containing 10% fetal bovine serum (v / v) and 1% double antibody (v / v), and statically culture in a 37°C, 5% CO2 humidified incubator until the density of HepG-2 cells is about 8×10 4 / mL. Then, treat HepG-2 cells with Se / ZIF8 (prepared in Example 1) at final system concentrations (the concentration after adding the drug to the cell culture plate) of 0, 1.25 μg / mL, 2.5 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL, and 40 μg / mL for 24 h, and detect the survival rate of HepG-2 cells using an MTT assay kit according to Example 2.

[0064] II. Experimental Results The results are as Figure 5As shown, the effect of Se / ZIF8 on the viability of HepG-2 cells showed concentration dependence. The cell viability corresponding to Se / ZIF8 with a concentration not exceeding 1.25 μg / mL was basically similar to that of the control group. Therefore, the concentration of Se / ZIF8 was selected as 0.125 - 1 μg / mL for the tumor killing experiment.

[0065] Example 4 Anti-hepatocarcinoma cell activity of Se / ZIF8 in combination with NK cells in vitro I. Experimental method HepG-2 cells were inoculated into a cell culture plate and added with DMEM complete medium containing 10% fetal bovine serum (v / v) and 1% double antibody (v / v). They were statically cultured in a humidified incubator at 37°C and 5% CO2 until the density of HepG-2 cells reached about 8×10 4 / mL.

[0066] NK-92 cells were inoculated into a cell culture plate and added with α-MEM complete medium containing 10% fetal bovine serum (v / v), 10% horse serum (v / v) and 1% double antibody (v / v). They were statically cultured in a humidified incubator at 37°C and 5% CO2 until the density of NK-92 cells reached about 1×10 7 / mL.

[0067] The ZIF8, intermediate and Se / ZIF8 prepared in Example 1 were used to treat the cells respectively according to the following methods: Method I: NK-92 cells were pretreated with 1 μg / mL ZIF8, intermediate or Se / ZIF8 for 24 h, and then used to treat HepG-2 cells for 24 h; Method II: HepG-2 cells were pretreated with 1 μg / mL ZIF8, intermediate or Se / ZIF8 for 24 h, and then NK-92 cells were added to treat HepG-2 cells for 24 h; Method III: 1 μg / mL ZIF8, intermediate or Se / ZIF8 and NK-92 cells were added to HepG-2 cells simultaneously and treated for 24 h.

[0068] According to the method of Example 2, an MTT kit was used to detect the viability of HepG-2 cells after being treated by the three methods.

[0069] II. Experimental results The results are as Figure 6As shown, at a concentration of 1 μg / mL, after pretreatment of HepG-2 cells with the intermediate and Se / ZIF8 for 24 h and then adding NK cells, both can significantly enhance the killing effect of NK cells on HepG-2 cells, and there is no significant difference in the sensitizing effect of the two on NK cells, while using ZIF8 alone has no such effect. In addition, modifying the intermediate (i.e., Se / ZIF8) with DSPE-PEG2000 can enhance the stability of the material and the metabolism time of the drug ( Figure 7 ), so Se / ZIF8 was selected for the tumor killing experiment.

[0070] The effects of Method I and Method III on enhancing the killing effect of NK cells on HepG-2 cells are not as significant as Method II. This is attributed to the sequential regulation of Se / ZIF8 on tumor cells and NK cells. Therefore, Method II was used for the tumor killing experiment, that is, HepG-2 cells were pretreated with Se / ZIF8 for 24 h and then NK cells were added.

[0071] Example 5 Effects of nanoparticles of different selenium forms alone or in combination with NK cells on the survival rate of HepG-2 cells I. Experimental method HepG-2 cells and NK-92 cells were prepared according to Example 4.

[0072] LNT-SeNPs: Take 1 mL of lentinan (LNT, CAS No.: 37339-90-5) solution (2 mg / mL), 500 μL of Na2SeO3 solution (100 mM), and 500 μL of vitamin C solution (400 mM) and add them to 8 mL of ultrapure water. Stir and react at room temperature for 24 h to obtain.

[0073] Selenocysteine (SeC2): Purchased from Shanghai Yuanye Bio-Technology Co., Ltd.

[0074] LNT-SeNPs+ZIF8: Prepare LNT-SeNPs according to the above method, and then mix them with ZIF8 prepared in Example 1.

[0075] SeC2+ZIF8: Mix SeC2 with ZIF8 prepared in Example 1.

[0076] Single action: Uniformly at a final concentration (the concentration when co-cultured with cells) containing 0.2 μg / mL of Se and 0.8 μg / mL of ZIF8, treat HepG-2 cells with LNT-SeNPs, SeC2, LNT-SeNPs+ZIF8, SeC2+ZIF8, ZIF8 (prepared in Example 1), and Se / ZIF8 (prepared in Example 1) for 24 h.

[0077] Combined action: Uniformly according to the Se content of 0.2 μg / mL and the ZIF8 concentration of 0.8 μg / mL in the final concentration (the concentration when co-cultured with cells), pre-treat HepG-2 cells with LNT-SeNPs, SeC2, LNT-SeNPs+ZIF8, SeC2+ZIF8, ZIF8 (prepared in Example 1), and Se / ZIF8 (prepared in Example 1) for 24 h, and then add NK-92 cells to treat HepG-2 cells for 24 h.

[0078] II. Experimental results The results are shown in Table 1 and Figure 8 As shown, when LNT-SeNPs, SeC2, LNT-SeNPs+ZIF8, SeC2+ZIF8, ZIF8, and Se / ZIF8 act alone on HepG-2 cells, there is no obvious killing effect, while the combined action of Se / ZIF8 and NK cells shows a significant killing effect on HepG-2 cells, playing a synergistic effect.

[0079] Table 1 Effects of different materials alone or in combination with NK cells on the survival rate of HepG-2 cells (%)

[0080] Example 6 In vitro anti-simulated solid tumor activity of Se / ZIF8 synergistic with NK cells I. Experimental method Since NK cells aggregate on the surface of tumor spheres and it is difficult to observe the actual size of the spheres, a tumor sphere model was constructed using HepG-2 stably transfected with GFP. HepG-2 tumor spheres were formed and grown in a 96-well plate (Corning merge) with an ultra-low adhesion surface: Each well was inoculated with HepG-2 cells stably transfected with GFP at a density of 1000 cells / well. After the HepG-2 tumor spheres grew for 3 days, 2 μg / mL of Se / ZIF8 prepared in Example 1 was added to pre-treat the HepG-2 tumor spheres for 24 h. After 24 h, NK-92 cells (1×10 6 / well) were added and incubated for 7 days. During the incubation process, fluorescence microscopy was used to take pictures of HepG-2 tumor spheres at different depths.

[0081] II. Experimental results The results are as Figure 9 shown. When Se / ZIF8 and NK cells act alone, the volume of the tumor spheres decreases slightly. After the combined treatment, the volume of the spheres further decreases significantly, indicating that Se / ZIF8 synergistic with NK cells can enhance the tumor killing effect.

[0082] Example 7 Inhibitory effect of the combined action of Se / ZIF8 and NK cells on tumor growth in an orthotopic liver cancer-bearing mouse model I. Experimental methods Inject 10 μL of serum-free medium containing 2×10 5 HepG-2 cells into the left lobe of the liver of male BALB / c nude mice to establish an orthotopic liver tumor model.

[0083] Seven days after tumor inoculation, the mice were randomly divided into 4 groups (n = 8) and treated as follows: Se / ZIF8 group: Prepare the Se / ZIF8 obtained in Example 1 with normal saline and inject 2 mg / kg of Se / ZIF8 into the caudal vein of the mice, respectively on days 1, 3, 5, 7, 9, 11, and 13.

[0084] NK-92 cell group: Inject 2×10 7 cells / mouse of NK-92 cells (prepared with serum-free medium) into the caudal vein of the mice, respectively on days 2, 4, 6, 8, 10, 12, and 14.

[0085] Se / ZIF8 + NK-92 cell group: The administration concentrations of Se / ZIF8 and NK-92 cells are the same as those in the Se / ZIF8 group and the NK-92 cell group. Inject Se / ZIF8 into the caudal vein on days 1, 3, 5, 7, 9, 11, and 13, and inject NK-92 cells into the caudal vein on days 2, 4, 6, 8, 10, 12, and 14.

[0086] Control group: Administer an equal amount of normal saline once a day for 14 consecutive days.

[0087] During the experiment, the development of liver tumors in each group was observed and evaluated by animal magnetic resonance imaging (MRI). At the end of the experiment, the mice were sacrificed, the livers were surgically dissected for measurement, and the tumor masses were dissected and weighed.

[0088] II. Experimental results The results of magnetic resonance imaging are as Figure 10 shown. On the 0th day of treatment, the MRI image clearly showed tumors (white dotted frames) on the livers of the orthotopic liver tumor mice, while there were no tumors in the normal mice. After 14 days of treatment, the liver tumors in the control group, Se / ZIF8 group, and NK-92 cell group developed rapidly, but the treatment of the Se / ZIF8 + NK-92 cell group could significantly inhibit the growth of liver tumors, and the tumor area was significantly smaller than that of other groups (white dotted frames), indicating that Se / ZIF8 synergistically with NK cells has excellent antitumor effects.

[0089] At the end of the 14th day of treatment, the mice were dissected, and the livers were taken for measurement and analysis ( Figure 11). The digital photos of the liver clearly show the difference between the tumor and the liver. The livers of the mice in the control group, Se / ZIF8 group, and NK-92 cell group were swollen and progressed to a serious tumor size (white dotted line), further indicating that neither Se / ZIF8 alone nor NK-92 cells alone could effectively inhibit tumor growth. However, the treatment with Se / ZIF8 + NK-92 cells effectively prevented the malignant proliferation of the tumor, and the tumor volume was significantly smaller than that of other groups, with less liver lesions.

[0090] The digital photos of the tumors of the mice at the end of the treatment showed that the tumor volume in the Se / ZIF8 + NK-92 cell group was significantly reduced, in sharp contrast to other treatments. The tumor volumes in the Se / ZIF8 group and the NK group after monotherapy were both smaller than those in the control group, but the differences were not significant ( Figure 12 ). The average tumor weight in the control group was about 1.2875 g, and the average tumor weights in the Se / ZIF8 group and the NK-92 cell group were 1.0193 g and 0.9013 g respectively, which were 79% and 70% of the control group respectively. The average tumor weight in the Se / ZIF8 + NK-92 cell group was only 0.4184 g, about 32% of the control group ( Figure 13 ). The tumor inhibition rate in the Se / ZIF8 + NK-92 cell group was as high as 67%, far exceeding 21% in the Se / ZIF8 group and 30% in the NK-92 cell group ( Figure 14 ). In summary, the combination of Se / ZIF8 and NK cells has excellent anti-tumor effects on liver tumors in vivo.

[0091] The above embodiments are the preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. Use of a composition in the preparation of a drug for treating liver cancer, characterized in that: The composition comprises selenium-based nanoparticles Se / ZIF8 and NK cells; The preparation method of the selenium-based nanoparticles Se / ZIF8 comprises the following steps: fully reacting selenium powder and zeolite imidazolate framework material ZIF8 to obtain the selenium-based nanoparticles Se / ZIF8.

2. The use according to claim 1, characterized in that: The preparation method of the selenium-based nanoparticles Se / ZIF8 comprises the following steps: fully reacting selenium powder and zeolite imidazolate framework material ZIF8, and then modifying the product with DSPE-PEG2000 to obtain the selenium-based nanoparticles Se / ZIF8.

3. The use according to claim 1, characterized in that: The usage ratio of the selenium powder and the zeolite imidazolate framework material ZIF8 is 1 g: (3-5) g.

4. The use according to claim 1, characterized in that: The temperature of the sufficient reaction is 255-265°C.

5. The use according to claim 1, characterized in that: The time for the full reaction is 0.8 to 1.2 h.

6. The use according to claim 2, characterized in that: The usage ratio of the product to DSPE-PEG2000 is 1 g: (0.8-1.2) g.

7. The use according to claim 1, characterized in that: The dosage ratio of the selenium-based nanoparticles Se / ZIF8 and NK cells in the composition is (0.125-1) μg: (0.8-1.2)×10 7 indivual.

8. The use according to any one of claims 1 to 7, characterized in that: The liver cancer is primary liver cancer.

9. The use according to claim 8, characterized in that: The primary liver cancer is hepatocellular carcinoma.

10. A drug for treating liver cancer, characterized in that: The medicine comprises the composition according to claim 1.

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