WO2024198628A1 - Pharmaceutical composition and use thereof in preparation of drug for treating tumors - Google Patents

Abstract

Provided is a pharmaceutical composition comprising a metal oxide nanoparticle and a soluble dietary fiber, wherein the metal oxide nanoparticle is selected from one or more of a tungsten trioxide nanoparticle, a titanium dioxide nanoparticle, a manganese dioxide nanoparticle and a molybdenum oxide nanoparticle. Further provided is the use of the pharmaceutical composition in the preparation of a drug for treating tumors. The provided pharmaceutical composition has a significant inhibitory effect on tumors, and slows down an increase rate of tumor volume, and/or reduces the tumor volume and weight. In addition, the pharmaceutical composition in combination with a tumor vaccine and an immune checkpoint inhibitor can further enhance the tumor inhibitory effect. The provided pharmaceutical composition has the advantages of readily available raw materials for preparation, a simple preparation method and low costs, and is suitable for large-scale production and has good application prospects.

Description

A pharmaceutical composition and its use in preparing a drug for treating tumors Technical Field

The present invention relates to the field of medicine, and in particular, to a pharmaceutical composition and use thereof in preparing an anti-tumor drug.

Background Art

Malignant tumors are one of the serious diseases that endanger human health. The number of cancer cases in my country is about 1.7 million per year, and the incidence rate has been rising in recent years. The number of patients who die from cancer each year is 1.5 million, surpassing cardiovascular and cerebrovascular diseases to become the first cause of death ["China Modern Medicine Journal", Zhang Yangde and Peng Jian; 2003, No. 10, pp. 43-46]. Although the diagnostic technology level of tumors in my country has made great progress, the treatment mainly relies on surgery, chemotherapy, radiotherapy, and immunotherapy. These therapies have great side effects on the human body and cause unbearable pain to patients, seriously affecting the quality of life of patients.

Summary of the invention

In order to solve the above technical problems, the present invention provides a pharmaceutical composition, which has a significant inhibitory effect on tumors and does not cause additional pain to patients.

Therefore, on the one hand, the present invention provides a pharmaceutical composition, characterized in that the pharmaceutical composition comprises metal oxide nanoparticles and soluble dietary fiber; the metal oxide nanoparticles are selected from one or more of tungsten trioxide nanoparticles, titanium dioxide nanoparticles, manganese dioxide nanoparticles, and molybdenum oxide nanoparticles.

On the other hand, the present invention also provides the use of the pharmaceutical composition provided in the present application in the preparation of anti-tumor drugs.

As shown in the experimental results, the pharmaceutical composition provided by the present invention has a significant inhibitory effect on tumors, slowing down the rate of increase in tumor volume, and/or reducing the volume and weight of the tumor; and when used in combination with tumor vaccines and immune checkpoint inhibitors, it can further enhance the tumor inhibition effect. In a preferred embodiment, the composition is made into particles, the metal oxide nanoparticles are the core, and the The soluble dietary fiber is the outer shell; in this preferred embodiment, after the pharmaceutical composition is ingested into the body, the soluble dietary fiber is degraded in the large intestine, and then the inner core metal oxide nanoparticles are exposed and infiltrate into the intestinal mucosa to exert their biological functions and reduce their side effects in other parts of the body.

The pharmaceutical composition provided by the present invention has readily available raw materials for preparation, a simple preparation method, low cost, is suitable for large-scale production, and has good application prospects.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an electron microscope characterization image of β-glucan-coated tungsten trioxide nanoparticles (abbreviated as WO ₃ @Glu NP) prepared in Example 1 of the present invention.

Figures 2A, 2B and 2C show the melanoma volume detection results of each mouse at different times in the control group, the group treated with tungsten trioxide and β-glucan (abbreviated as _WO3NP +Glu) and the group treated with _WO3 @GluNP; Figure 2D shows the result of averaging the volume data of each group of mice and plotting them in the same figure; Figure 2E shows the tumor weight in tumor-bearing mice in the control group, the group treated with _WO3NP +Glu and the group treated with _WO3 @GluNP; Figure 2F shows the percentage of T cells in TIL (TIL is the abbreviation of tumor-infiltrating lymphocytes) in the control group, the group treated with _WO3NP +Glu and the group treated with _WO3 @GluNP.

Figures 3A, 3B and 3C show the melanoma volume detection results of each mouse at different times in the control group, the group treated with immune checkpoint inhibitors (abbreviated as ICI) and the group treated with ICI+ _WO3 @Glu NP; Figure 3D shows the results of averaging the volume data of each group of mice and plotting them in the same figure; Figure 3E shows the tumor weight in tumor mice in the control group, the group treated with ICI and the group treated with ICI+ _WO3 @Glu NP; Figure 3F shows the percentage of T cells in TIL (TIL is the abbreviation of tumor-infiltrating lymphocytes) in the control group, the group treated with ICI and the group treated with ICI+WO3@Glu NP; Figure 3G shows CD4 and CD8 staining sections; Figures 3H and 3I show the changes in the content of CD4+T cells and CD8+T cells in the sections, respectively.

FIG. 4 shows the results of small animal in vivo imaging of pancreatic cancer mice in the control group, the ICI-combination treatment group (anti-PD-1 antibody and anti-CTLA-4 antibody were used in combination), and the ICI-combination + WO ₃ @Glu NP treatment group at different times.

Figure 5 shows the control group, the ICI-combined treatment group and the ICI-combined + WO ₃ @Glu NP treatment group. Photo of pancreatic tissue of pancreatic cancer mice in the treatment group 14 days later.

FIG. 6 shows the results of small animal in vivo imaging of colorectal cancer mice in the control group, the ICI-combination treatment group, and the ICI-combination + WO ₃ @Glu NP treatment group.

DETAILED DESCRIPTION

The invention provides a pharmaceutical composition, characterized in that the pharmaceutical composition comprises metal oxide nanoparticles and soluble dietary fiber; the metal oxide nanoparticles are selected from one or more of tungsten trioxide nanoparticles, titanium dioxide nanoparticles, manganese dioxide nanoparticles, and molybdenum oxide nanoparticles.

The composition of the present application can be prepared into an aqueous solution, a suspension, an emulsion or a mixture in any form.

There is no particular limitation on the soluble dietary fiber, but commonly used soluble dietary fiber is preferred. For example, the soluble dietary fiber is selected from one or more of β-glucan, lignin, resistant starch, cellulose, inulin, hemicellulose, lactulose, wheat dextrin, and galacto-oligosaccharide.

The metal oxide nanoparticles and soluble dietary fiber in the pharmaceutical composition can be used after simple mixing.

In order to obtain a better anti-tumor effect, in a preferred embodiment, the pharmaceutical composition has a core-shell structure; the metal oxide nanoparticles are the core and the soluble dietary fiber is the shell. In this way, when the pharmaceutical composition is ingested into the body, the soluble dietary fiber is degraded in the large intestine, and then the metal oxide nanoparticles in the core are exposed and infiltrate into the intestinal mucosa to exert their biological functions and reduce the side effects of the metal oxide nanoparticles in other parts of the body.

The particle size of the pharmaceutical composition having a core-shell structure is not particularly limited as long as it is nanometer-scale; however, in a preferred embodiment, the diameter of the inner core is 20-100 nanometers, preferably 30-60 nanometers; in another preferred embodiment, the thickness of the outer shell is 50-350 nanometers, preferably 80-200 nanometers; to facilitate absorption and timely degradation of soluble dietary fiber.

The diameter of the core and the thickness of the shell of the present application can be measured by a dynamic light scattering method.

In the present application, the mass ratio of the nano metal oxide to the soluble dietary fiber is not particularly limited, but in order to obtain a better tumor inhibition effect, in a preferred embodiment, the mass ratio of the metal oxide to the soluble dietary fiber is 1:1-100, such as 1:95, 1:90, 1:85; preferably 1:2-80, such as 1:75, 1:70, 1:65; more preferably 1:3-60, such as 1:55; More preferably, it is 1:5-50, for example, 1:45, 1:40, 1:35, 1:30, 1:25, 1:20, 1:15, 1:10, 1:8 and the like.

In order to obtain better anti-tumor effects, the drug combination is used to assist in tumor immunotherapy that enhances the body's innate immunity and specific immune response.

In a preferred embodiment, the pharmaceutical composition further comprises one or more of a tumor vaccine and an immune checkpoint inhibitor.

The tumor vaccine is not particularly limited. Preferably, the tumor vaccine includes one or more of: whole cell vaccine, tumor polypeptide vaccine, genetic engineering vaccine and antibody tumor vaccine.

The checkpoint inhibitor is not particularly limited. Preferably, the checkpoint inhibitor is selected from one or more of anti-PD-1 monoclonal antibodies, anti-PD-L1 monoclonal antibodies, and anti-CTLA-4 monoclonal antibodies.

The core-shell structured pharmaceutical composition of the present invention can be prepared by various methods, such as precipitation method, chemical adsorption method, chemical deposition method, and microemulsion method.

When the soluble dietary fiber is β-glucan, preferably, the core-shell structured pharmaceutical composition of the present invention is prepared by a method comprising the following steps:

1) preparing an alkaline solution of β-glucan: mixing β-glucan and an emulsifier, and adjusting the pH value to 8-10;

2) preparing an acid solution of the metal oxide nanoparticles: mixing the metal oxide nanoparticles and a weak acid;

3) At room temperature, the acid solution of the metal oxide nanoparticles is added dropwise into the alkaline solution of the soluble dietary fiber under stirring to adjust the pH to 4-6, and then the stirring is continued for 0.5-2 hours.

Preferably, washing and drying are performed in step 3).

Preferably, the room temperature includes a temperature of 20-45°C, such as 20°C, 25°C, 30°C, 35°C, 40°C or 45°C, etc.; the reaction time can be 40min, 50min, 70min, 80min, 90min, 100min, etc.

In a preferred embodiment, the weak acid is one or more of acetic acid, oxalic acid, formic acid and propionic acid.

In a preferred embodiment, the emulsifier may be one or more of Tween 80, Tween 60, polyglycerol fatty acid ester, and pectin;

In the present application, the tumor also includes a tumor that was not completely removed after tumor resection.

In the present application, the tumor includes a tumor that re-emerges after tumor clearance.

In the present application, the tumor includes solid tumors and non-solid tumors.

The present invention also provides the use of the pharmaceutical composition in the preparation of a drug for treating tumors; preferably, the tumor is selected from breast tumors, colon tumors, liver tumors, stomach tumors, kidney tumors, pancreatic cancer, colorectal cancer, ovarian tumors, lymphomas, osteosarcomas, gliomas, prostate cancer or melanoma.

The pharmaceutical composition of the present application will not substantially induce systemic inflammatory response.

The drug combination of the present application does not substantially induce hemolytic reactions, heart damage, liver damage, spleen damage, lung damage and/or kidney damage.

The present invention also provides a method for treating tumors, which comprises administering the pharmaceutical composition provided by the present application to a tumor patient, for example, orally. The dosage of the pharmaceutical composition provided by the present invention can be taken daily, for example, using the formula m = user's body weight (kg) × (100-500) mg/kg, and can be taken 1-3 times a day.

The present invention also provides a method for treating tumors that may include the combined administration of other tumor therapeutic drugs, such as tumor vaccines and immune checkpoint inhibitors. Tumor vaccines and immune checkpoint inhibitors may be administered by intraperitoneal injection at a dose of 50-250 μg/time.

Example 1: Preparation of a mixture of tungsten trioxide nanoparticles (purchased from Sigma-Aldrich (USA)) and β-glucan (abbreviated as WO ₃ NP+Glu)

3 mg of tungsten trioxide nanoparticles and 3 mg of β-glucan were mixed evenly, wherein the weight ratio of β-glucan to tungsten trioxide nanoparticles was 1:1.

Example 2: Preparation of β-glucan-coated tungsten trioxide nanoparticles (abbreviated as WO ₃ @Glu NP)

The preparation method of WO3@Glu NP is as follows:

(1) preparing a dextran base solution: uniformly mixing 20 ml of an aqueous solution of 0.1 wt % β-glucan and 0.1 wt % Tween 80, and adjusting the pH of the solution to 11 with 2 wt % sodium hydroxide;

(2) Preparation of an acid solution of tungsten trioxide nanoparticles: 5 mg of tungsten trioxide nanoparticles (purchased from Sigma-Aldrich, USA) were added to 5 ml of a 4 wt % acetic acid solution;

(3) At room temperature, 5 ml of the acid solution of tungsten trioxide nanoparticles prepared in step (2) was added dropwise into 5 ml of the β-glucan base solution prepared in step (1), and the pH of the solution was adjusted to 5 with 4 wt % acetic acid, and then stirred for 1 hour;

(4) Wash with water and dry to obtain WO ₃ @Glu NPs.

The electron microscope characterization of the obtained WO ₃ @Glu NP is shown in Figure 1, where the left picture is tungsten trioxide nanoparticles (abbreviated as WO ₃ NP) and the right picture is WO ₃ @Glu NP. The weight ratio of β-glucan and tungsten trioxide nanoparticles is 1:1, the average diameter of tungsten trioxide nanoparticles is 46nm, and the average thickness of β-glucan shell membrane is 100nm.

The following experiments were conducted using the WO ₃ NP+Glu prepared in Example 1 and the WO ₃ @Glu NP prepared in Example 2.

Example 3: Experiment on the inhibition of melanoma by WO ₃ NP+Glu and WO ₃ @Glu NP

Since patients undergoing long-term tumor immunotherapy usually have intestinal disorders, the mouse model used in this application uses a tumor and enteritis superimposed model.

Experimental animal modeling: 0.1 mL (1×10 ⁶ ) B16 tumor cells were injected subcutaneously into the back of 6-8 week old C57BL/6 female mice. When the tumor volume of the mice reached 150 mm ³ , they were randomly divided into groups, with 5 mice in each group. Dextran sulfate sodium (DSS) was added to drinking water to simulate clinical immune-related enteritis (DSS concentration 2% by weight, administered for 5 days); DSS was directly added to drinking water, and mice drank water for 5 days without quantitative requirements.

Drug administration: Starting from the 0th day of enteritis modeling, drugs or water were administered every other day as follows (once a day): 0.1 ml ultrapure water/mouse/day (oral, as the control group, abbreviated as Ctrl), 3 mg WO ₃ NP+Glu/mouse/day (oral, WO ₃ NP+Glu group), 3 mg WO ₃ @Glu NP/mouse/day (oral, WO ₃ @Glu NP group).

Each group of mice was fed for 14 days, and the volume of the tumor was measured every other day. The test results are shown in Figures 2A, 2B, 2C, and 2D. After 14 days of feeding, the mice were killed, and the weight of the tumor and tumor-infiltrating T cells were measured. The test results are shown in Figures 2E and 2F, respectively.

Tumor volume detection method: Use a vernier caliper to measure the length of the major axis and minor axis of the tumor tissue, and calculate according to the formula V = 0.5 × ab ² , where a represents the major axis and b represents the minor axis.

Tumor weight detection method: After killing the mice, the tumor tissue was separated and weighed.

Tumor-infiltrating T cell detection method: After killing mice, the tumor tissue was ground into a single-cell suspension, stained with CD45 fluorescent antibody (purchased from Beijing Chengzhi Kewei Biotechnology Co., Ltd.) and CD3 fluorescent antibody (purchased from Beijing Chengzhi Kewei Biotechnology Co., Ltd.), and the fluorescence signal was detected by flow cytometry.

Figures 2A, 2B and 2C show the tumor volume of each mouse in the control group, the WO ₃ NP+Glu and WO ₃ @Glu NP treatment groups at different times. Since one mouse died in the control group during the experiment, the data of 4 mice were used in each group when drawing the graphs.

Figure 2D shows the volume data of each group of mice being averaged and plotted in the same figure. It can be seen from the figure that 8 days after enteritis modeling, the tumor volumes of the control group, the group treated with WO ₃ NP+Glu and the group treated with WO ₃ @Glu NP gradually showed significant differences. Both WO ₃ NP+Glu and WO ₃ @Glu NP were able to inhibit the increase in tumor volume, and WO ₃ @Glu NP showed better tumor inhibition effect than WO ₃ NP+Glu.

It can also be seen from Figure 2E that both WO ₃ NP+Glu and WO ₃ @Glu NP can inhibit the increase of tumor weight, and WO ₃ @Glu NP shows better tumor inhibition effect than WO ₃ NP+Glu.

As can be seen from FIG. 2F , both WO ₃ NP+Glu and WO ₃ @Glu NP can increase the percentage of T cells in TIL (TIL is the abbreviation of tumor-infiltrating lymphocytes), and WO ₃ @Glu NP shows a better effect than WO ₃ NP+Glu.

Example 4: Experiment on the inhibition of melanoma by administration of WO ₃ @Glu NP combined with immune checkpoint inhibitor (ICI)

Since patients undergoing long-term tumor immunotherapy usually have intestinal disorders, the mouse model used in this application uses a tumor and enteritis superimposed model.

Experimental animal modeling: 0.1 mL (1×10 ⁶ ) B16 tumor cells were injected subcutaneously into the back of 6-8 week old C57BL/6 female mice. When the tumor volume of the mice reached 150 mm ³ , they were randomly divided into groups, with 5 mice in each group. Dextran sulfate sodium (DSS) was added to drinking water to simulate clinical immune-related enteritis (DSS concentration 2% by weight, administered for 5 days); DSS was directly added to drinking water, and mice drank water for 5 days without quantitative requirements.

Drug administration: Starting from day 0 of enteritis modeling, the mice were treated every other day according to the following regimen (once a day): 0.1 ml ultrapure water/mouse/day (oral, control group, abbreviated as Ctrl), 200 μg anti-PD-1 antibody/mouse/day (intraperitoneal injection, ICI-treated group), 3 mg WO ₃ @Glu NP (oral) + 200 μg anti-PD-1 antibody (intraperitoneal injection)/mouse/day (ICI+WO ₃ @Glu NP-treated group).

Each group of mice was fed for 14 days, and the volume of the tumor was measured every other day. The test results are shown in Figures 3A, 3B, 3C, and 3D. After 14 days of feeding, the mice were killed, and the weight of the tumor and tumor-infiltrating T cells were measured. The test results are shown in Figures 3E and 3F, respectively.

Tumor volume detection method: Use a vernier caliper to measure the length of the major axis and minor axis of the tumor tissue, and calculate according to the formula V = 0.5 × ab ² , where a represents the major axis and b represents the minor axis.

Tumor weight detection method: After killing the mice, the tumor tissue was separated and weighed.

Tumor-infiltrating T cell detection method: After killing mice, the tumor tissue was ground into a single-cell suspension, stained with CD45 fluorescent antibody (purchased from Beijing Chengzhi Kewei Biotechnology Co., Ltd.) and CD3 fluorescent antibody (purchased from Beijing Chengzhi Kewei Biotechnology Co., Ltd.), and the fluorescence signal was detected by flow cytometry.

Tumor tissue sectioning and staining: Another part of the tumor was fixed and embedded in paraffin for sectioning. Immunohistochemical antibodies were used to stain CD4 and CD8 to detect changes in the content of CD4+T cells and CD8+T cells. The results are shown in Figures 3G, 3H, and 3I.

Figures 3A, 3B and 3C show the results of tumor volume detection of each mouse at different times in the control group, ICI treatment group and ICI+WO ₃ @Glu NP treatment group. Since one mouse died in the control group during the experiment, the data of 4 mice were used in each group when drawing the graphs.

Figure 3D is the average volume data of each group of mice plotted in the same figure. It can be seen from the figure that 8 days after enteritis modeling, the tumor volumes of the control group, the ICI-treated group and the ICI+WO ₃ @Glu NP-treated group gradually showed significant differences. Both ICI and ICI+WO ₃ @Glu NP were able to inhibit the increase in tumor volume, and ICI+WO ₃ @Glu NP showed better tumor inhibition effect than ICI.

It can also be seen from Figure 3E that both ICI and ICI+WO ₃ @Glu NPs can inhibit the increase of tumor weight, and ICI+WO ₃ @Glu NPs show better tumor inhibition effect than ICI.

As can be seen from Figure 3F, both ICI and ICI+WO ₃ @Glu NPs can increase the percentage of T cells in TIL (TIL is the abbreviation of tumor-infiltrating lymphocytes), and ICI+WO ₃ @Glu NPs show better effects than ICI.

As can be seen from Figure 3G, there are more staining marks in the ICI+WO ₃ @Glu NP than in ICI in the sections, indicating that the content of CD4+T cells and CD8+T cells in the tumor tissues of mice treated with ICI+WO ₃ @Glu NP is higher, which means that WO ₃ @Glu NP adjuvant therapy improves the infiltration of immune cells. The numerical values of CD4+T cells and CD8+T cells quantification are shown in Figures 3H and 3I.

From the above data, it can be seen that WO ₃ @Glu NP and ICI may exert different anti-tumor effects, and the combination of WO ₃ @Glu NP and ICI can further enhance the anti-tumor effect.

Example 4: Experiment on inhibiting the development of in situ pancreatic cancer by administering WO ₃ @Glu NP in combination with anti-PD-1 antibody and anti-CTLA-4 antibody

Experimental animal modeling: 0.025mL (1×10 ⁶ ) luci-Panc02 tumor cells and (1×10 ⁶ ) mouse pancreatic stellate cells were injected orthotopically into the pancreas of 6-8 week old C57BL/6 female mice. Five days after tumor implantation, the mice were randomly divided into groups, with 6 mice in each group. At this time, dextran sulfate sodium (DSS) was added to the drinking water to simulate clinical immune-related enteritis (DSS concentration 2% by weight, administered for a total of 9 days); DSS was directly added to the drinking water, and the mice drank water for 9 days without quantitative requirements.

Dosing treatment: Starting from day 0 of enteritis modeling, the mice were treated every other day according to the following scheme (once a day): 0.1 ml ultrapure water/mouse/day (oral, control group, abbreviated as Ctrl), (100 μg anti-PD-1 antibody + 100 μg anti-CTLA-4 antibody)/mouse/day (intraperitoneal injection, ICI-combined treatment group); (100 μg anti-PD-1 antibody + 100 μg anti-CTLA-4 antibody + 3 mg WO3@Glu NP)/mouse/day (intraperitoneal injection of antibodies, oral administration of nanoparticles, ICI-combined + WO ₃ @Glu NP treatment group).

Each group of mice was raised for 14 days, and small animal live imaging was performed on the 5th, 8th, 11th and 14th days of enteritis modeling. The test results are shown in Figure 4. After 14 days of raising, the mice were killed, the spleen and pancreas were removed, and the size of the pancreas was detected. The test results are shown in Figure 5.

Small animal in vivo imaging: D-luciferin potassium salt was intraperitoneally injected at a dose of 150 mg/kg body weight, and 3% sodium pentobarbital solution was intraperitoneally injected 10 minutes later. Small animal optical 3D in vivo imaging system (IVIS-Spectrum) (manufacturer PerkinElmer) was then used for small animal in vivo imaging.

It can be clearly seen from Figure 4 that as the feeding time of the three groups of mice increases, the tumors gradually worsen (the fluorescence intensity scale indicates the order of magnitude of tumor cells from low to high, from 2*10 ⁴ to 5*10 ⁵ ). The fastest growing mice are those in the control group, followed by the mice in the ICI-combination treatment group, and the tumor growth rate of the mice in the ICI-combination + WO ₃ @Glu NP treatment group is the slowest. From the 11th day to the 14th day, the tumors of the mice in the control group and the ICI-combination treatment group still grow rapidly; while the tumor changes of the mice in the ICI-combination + WO ₃ @Glu NP treatment group are not obvious. This result shows that WO ₃ @Glu NP significantly promotes the inhibition of tumor growth by ICI-combination.

It can also be clearly seen from Figure 5 that at 14 days, the pancreases of the 6 mice in the control group and the ICI-combination treatment group were very large, and there was almost no difference; while the pancreases of the 6 mice in the ICI-combination + WO ₃ @Glu NP treatment group were significantly smaller than those in the control group and the ICI-combination treatment group. This result further illustrates that WO ₃ @Glu NP significantly promotes the efficacy of ICI-combination in inhibiting tumor growth.

Example 5: WO ₃ @Glu NPs were administered in combination with anti-PD-1 antibody and anti-CTLA-4 antibody to inhibit the growth of in situ colorectal cancer.

Experimental animal modeling: 0.025mL (1×10 ⁶ ) luci-CT 26 tumor cells were injected orthotopically into the cecum of 6-8 week old C57BL/6 female mice. Three days after tumor implantation, the mice were randomly divided into groups, with 6 mice in each group. At this time, dextran sulfate sodium (DSS) was added to the drinking water to simulate clinical immune-related enteritis (DSS concentration 2% by weight, administered for a total of 8 days); DSS was directly added to the drinking water, and the mice drank water for 8 days without quantitative requirements.

Dosing treatment: Starting from day 0 of enteritis modeling, the mice were treated every other day according to the following scheme (once a day): 0.1 ml ultrapure water/mouse/day (oral, control group, abbreviated as Ctrl), (100 μg anti-PD-1 antibody + 100 μg anti-CTLA-4 antibody)/mouse/day (intraperitoneal injection, ICI-combined treatment group); (100 μg anti-PD-1 antibody + 100 μg anti-CTLA-4 antibody + 3 mg WO3@Glu NP)/mouse/day (intraperitoneal injection of antibodies, oral administration of nanoparticles, ICI-combined + WO ₃ @Glu NP treatment group).

Each group of mice was raised for 14 days, and small animal live imaging was performed on the 5th, 8th and 11th day of enteritis modeling (the method is the same as that of Example 4). The test results are shown in Figure 6: As the feeding time of the three groups of mice increased, the tumors gradually worsened (the fluorescence intensity scale was from low to high, from 2*10 ⁴ to 5*10 ⁵ ); the tumors in the mice of the control group and the ICI-combined treatment group grew relatively fast, and a large number of tumor cells had appeared on the 8th day, as shown by a strong fluorescence signal. By the 11th day, the fluorescence intensity signal in the control group showed that the tumor volume increased significantly, while the tumor fluorescence signal area in other mice of the control group and the ICI-combined treatment group decreased slightly. This is because colorectal cancer is easy to metastasize, and the fluorescence statistics can no longer accurately show the tumor size, so subsequent small animal live imaging is no longer performed. On the 11th day, the tumors in the mice of the ICI-combined +WO ₃ @Glu NP treatment group were obviously smaller than the tumor size in the mice of the control group and the ICI-combined treatment group on the 8th day. This result indicated that _WO3 @Glu NPs significantly promoted ICI-combined inhibition of tumor growth.

Claims (10)

A pharmaceutical composition, characterized in that the pharmaceutical composition comprises metal oxide nanoparticles and soluble dietary fiber; the metal oxide nanoparticles are selected from one or more of tungsten trioxide nanoparticles, titanium dioxide nanoparticles, manganese dioxide nanoparticles, and molybdenum oxide nanoparticles; preferably, the soluble dietary fiber is selected from one or more of β-glucan, lignin, resistant starch, cellulose, inulin, hemicellulose, lactulose, wheat dextrin, and galacto-oligosaccharides. The pharmaceutical composition according to claim 1, characterized in that the pharmaceutical composition has a core-shell structure; the metal oxide nanoparticles are the core and the soluble dietary fiber is the shell. The pharmaceutical composition according to claim 2, characterized in that the diameter of the core is 20-100 nanometers, preferably 30-60 nanometers. The pharmaceutical composition according to claim 2, characterized in that the thickness of the shell is 50-350 nanometers, preferably 80-200 nanometers. The pharmaceutical composition according to claim 1, characterized in that the mass ratio of the metal oxide to the soluble dietary fiber is 1:1-100. The pharmaceutical composition according to claim 1, characterized in that the mass ratio of the metal oxide to the soluble dietary fiber is 1:5-50. The pharmaceutical composition according to any one of claims 1 to 6, characterized in that the pharmaceutical composition further comprises one or more of a tumor vaccine and an immune checkpoint inhibitor; preferably, the tumor vaccine comprises: one or more of a whole cell vaccine, a tumor polypeptide vaccine, a genetically engineered vaccine and an antibody tumor vaccine; preferably, the checkpoint inhibitor is selected from one or more of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody and an anti-CTLA-4 monoclonal antibody. The pharmaceutical composition according to claim 2, characterized in that the soluble dietary fiber is β-glucan, and the pharmaceutical composition is prepared by a method comprising the following steps: 1) preparing an alkaline solution of β-glucan: mixing β-glucan and an emulsifier, and adjusting the pH value to 8-10; preferably, the emulsifier is one or more of Tween 80, Tween 60, polyglycerol fatty acid ester, and pectin; 2) preparing an acid solution of the metal oxide nanoparticles: mixing the metal oxide nanoparticles and a weak acid; 3) at room temperature, dropwise add the soluble dietary fiber into the alkaline solution under stirring. The acid solution of the metal oxide nanoparticles is adjusted to pH 4-6, and then stirred for 0.5-2 hours. The pharmaceutical composition according to claim 8, characterized in that the weak acid is one or more of acetic acid, oxalic acid, formic acid, and propionic acid. Use of the pharmaceutical composition according to any one of claims 1 to 9 in the preparation of a drug for treating tumors; preferably, the tumor is selected from breast tumors, colon tumors, liver tumors, stomach tumors, kidney tumors, pancreatic cancer, colorectal cancer, ovarian tumors, lymphomas, osteosarcomas, gliomas, prostate cancer or melanoma.