Combined traditional Chinese medicine composition for treating tumors by improving tumor immune microenvironment and application of combined traditional Chinese medicine composition

Abstract

The invention relates to a combined traditional Chinese medicine composition for treating tumors by improving a tumor immune microenvironment and application thereof. The combined traditional Chinese medicine composition comprises rhein and hesperidin. Rheinic acid and hesperidin have stronger antitumor activity than single components in combined application, and the combined pharmaceutical composition can significantly inhibit the growth of cancer-related fibroblasts, reduce CCL2 secreted by cancer-related adipocytes and increase infiltration of CD8 + T cells at tumor sites, so that the growth of tumors is significantly inhibited, and the antitumor effect is improved. And a new method and thought are provided for combined treatment of cancers.

Description

Technical Field

[0001] The present invention belongs to the technical field of Chinese medicine compositions, and in particular relates to a combined Chinese medicine composition for treating tumors by improving the tumor immune microenvironment and its application. Background Art

[0002] The tumor immune microenvironment plays a key role in tumor development and treatment. In recent years, traditional Chinese medicine treatment has become an important means of tumor treatment. However, the impact of the tumor microenvironment on traditional Chinese medicine treatment is still an important research field. The occurrence and development of tumors are not only affected by the genetic variation of tumor cells themselves, but also regulated by the tumor immune microenvironment. In-depth research on the interaction between the tumor immune microenvironment and treatment is particularly important. Clarifying these contents is necessary for precise treatment based on the characteristics of the immune microenvironment of different tumor patients and improving the treatment effect. Cancer-associated fibroblasts and cancer-associated adipocytes, as two important stromal cells in the tumor immune microenvironment, are the main drivers of the immunosuppressive microenvironment that promotes tumor growth. Therefore, it is of great significance to find drugs and dosing regimens that simultaneously intervene in cancer-associated fibroblasts and cancer-associated adipocytes.

[0003] Rhein is the main component of the traditional Chinese medicine rhubarb. It has anti-cancer, antibacterial, anti-inflammatory, diuretic, laxative, and therapeutic effects on diabetic nephropathy. At the same time, it has a certain inhibitory effect on the fibrosis of breast cancer tumor tissue and renal fibrosis. Hesperidin is a flavonoid compound composed of the flavonoid hesperidin and a molecule of rutin. It has a wide range of anti-inflammatory, antioxidant, anti-diabetic, lipid-lowering, cardioprotective, liver-protective and neuroprotective effects. It can inhibit the production and activity of CCL2 in cancer-related fat cells, thereby inhibiting the development and metastasis of tumors.

[0004] In breast cancer, cancer-associated fibroblasts and cancer-associated adipocytes enhance the immunosuppressive microenvironment and promote the occurrence, development and metastasis of tumors. The combined use of rhein and hesperidin can inhibit breast cancer by removing cancer-associated fibroblasts and inhibiting the secretion of CCL2 by cancer-associated adipocytes. However, there are few detailed reports on the combined use of the two in tumor treatment and application. Summary of the invention

[0005] In view of the problems existing in the prior art, the present invention aims to use rhein and hesperidin in combination to inhibit the growth of cancer-related fibroblasts, reduce the secretion of CCL2 by cancer-related adipocytes, and increase CD8 + T cell infiltration has a significant effect in treating breast cancer. This is achieved through the following technical solutions: A combined Chinese medicine composition for treating tumors by improving the tumor immune microenvironment. The Chinese medicine composition comprises rhein and hesperidin.

[0006] Furthermore, rhein is rhein monomer or a pharmaceutically acceptable salt thereof, and hesperidin is hesperidin monomer or a pharmaceutically acceptable salt thereof.

[0007] Furthermore, the mass ratio of rhein to hesperidin is 1-10:1-10, and preferably the mass ratio of rhein to hesperidin is 1:1.

[0008] Furthermore, the dosage form of the combined Chinese medicine composition is any pharmaceutically acceptable dosage form, preferably nanoparticles, liposomes, nanoemulsions, microcapsules, microspheres, micropellets, solid dispersions, and more preferably nanoparticles.

[0009] The average particle size of the rhein and hesperidin co-loaded nanoparticles is preferably 50-400 nm, more preferably 100-300 nm.

[0010] The rhein and hesperidin co-loaded nanoparticles further include an emulsifier, an emulsifier, an organic phase and water. The emulsifier is preferably at least one of poloxamer 188, Tween 80, soybean lecithin, and polyoxyethylene hydrogenated castor oil EL35, and more preferably poloxamer 188. The emulsifier is preferably at least one of n-butanol, ethanol, polyethylene glycol 400, polyvinyl alcohol, and 1,2-propylene glycol, and more preferably polyvinyl alcohol. The oil is preferably at least one of methanol, acetonitrile, dichloromethane, chloroform, and ethyl acetate, and more preferably methanol.

[0011] The preparation method of nanoparticles of rhein and hesperidin is as follows: accurately weigh the prescribed amount of rhein, hesperidin and PLGA, dissolve them in an appropriate amount of organic phase, and dissolve them by ultrasonication in a water bath; use pure water to dissolve the prescribed amount of emulsifier and co-emulsifier as the aqueous phase; add the organic phase to the aqueous phase, ultrasonically emulsify in a water bath for 10 minutes, and remove the organic solvent by rotary evaporation under reduced pressure at 33°C to obtain nanoparticles co-loaded with rhein and hesperidin.

[0012] Furthermore, the Chinese medicine composition also includes any one or more combinations of pharmaceutical excipients that are pharmaceutically acceptable.

[0013] Furthermore, the pharmaceutical excipients include polylactic acid-glycolic acid copolymer, poloxamer 188 and polyvinyl alcohol.

[0014] Furthermore, the administration form of the Chinese medicine composition includes any one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration or transdermal administration, preferably intraperitoneal injection.

[0015] The use of a combined Chinese medicine composition in the preparation of anti-tumor drugs, wherein the tumors include melanoma, gastric cancer, lung cancer, pancreatic cancer, liver cancer, kidney cancer, breast cancer, cervical cancer, uterine cancer, skin cancer, prostate cancer, thyroid cancer, leukemia, lymphoma, esophageal cancer, oral cancer, intestinal cancer, nasal cancer, head and neck cancer, preferably breast cancer.

[0016] Application of a combined Chinese medicine composition in the preparation of a drug for improving the tumor immune microenvironment.

[0017] Furthermore, the immune microenvironment includes cancer-associated fibroblasts, cancer-associated adipocytes, tumor-associated macrophages, tumor-associated neutrophils, and tumor-infiltrating lymphocytes, preferably cancer-associated fibroblasts and cancer-associated adipocytes.

[0018] The traditional Chinese medicine composition provided by the present invention plays an important role in improving the tumor immune microenvironment. On the one hand, the nanoparticle dosage form effectively increases the solubility of rhein and hesperidin. On the other hand, the nanosized system can relieve the immunosuppressive effect in the tumor microenvironment, enhance the CD8+T cell infiltration in the tumor microenvironment, and significantly inhibit the growth of the tumor. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a transmission electron microscopy image of the nanoparticles co-loaded with rhein and hesperidin according to the present invention; Figure 2 This is a particle size distribution diagram of the nanoparticles co-loaded with rhein and hesperidin of the present invention; Figure 3 The stability test results of the nanoparticles co-loaded with rhein and hesperidin of the present invention; Figure 4 To investigate the in vitro release of the nanoparticles co-loaded with rhein and hesperidin of the present invention; Figure 5 The figure is a diagram showing the therapeutic effect of the nanoparticles co-loaded with rhein and hesperidin on the animal model of breast cancer in situ tumor; wherein A is a schematic diagram of the dosing regimen of the nanoparticles; B is the change in body weight of tumor-bearing mice during treatment; C is a diagram showing the change in tumor volume of tumor-bearing mice; D is the weight of the tumor peeled off from the tumor-bearing mice on the 14th day after drug treatment; E is a diagram showing the peeled off tumor from the tumor-bearing mice on the 14th day after different drug treatments; F is a diagram showing the change in tumor luminescence intensity of tumor-bearing mice; G is a curve of BLIrel ratios of different drug treatment groups at different times; and H is a diagram showing H&E staining of tumor tissue; Figure 6 Flow cytometry was used to detect the changes in immune cells and CCL2 expression in tumor tissues of tumor-bearing mice after treatment with different drugs; Figure 7 Immunofluorescence was used to detect α-SMA and CD8 in tumor tissues of tumor-bearing mice after treatment with different drugs. + Changes in T expression; Figure 8 H&E staining (bar 100 μm) and blood biochemical indexes of the main organs of tumor-bearing mice; Note: *P<0.05, **P<0.01, ***P<0.001. The t-test was used to evaluate the significant difference, and the results are expressed as mean ± SD. DETAILED DESCRIPTION

[0020] The present invention is further described below in conjunction with the accompanying drawings in order to better understand the technical solution.

[0021] Example 1: Preparation of rhein and hesperidin co-loaded nanoparticles The preferred ratio of rhein and hesperidin co-loaded nanoparticles in this embodiment is (rhein: hesperidin = 1:1) Rhein 5 mg, hesperidin 5 mg, PLGA 50 mg, poloxamer 188 300 mg, polyvinyl alcohol 100 mg, dilute to the required volume with water for injection.

[0022] The emulsified solvent diffusion method was used for preparation: the prescribed amount of rhein, hesperidin and PLGA were accurately weighed, dissolved in an appropriate amount of methanol, and dissolved by ultrasonication in a water bath to serve as the organic phase; the prescribed amount of poloxamer 188 (P188) and polyvinyl alcohol (PVA) were dissolved in pure water to serve as the aqueous phase; the organic phase was added to the aqueous phase, ultrasonically emulsified in a water bath for 10 minutes, and the organic solvent was removed by rotary evaporation under reduced pressure at 33°C to obtain rhein and hesperidin co-loaded nanoparticles.

[0023] Example 2: Preparation of rhein and hesperidin co-loaded nanoparticles The preferred ratio of rhein and hesperidin co-loaded nanoparticles in this embodiment is (rhein: hesperidin = 1:5) Rhein 1 mg, hesperidin 5 mg, PLGA 50 mg, poloxamer 188 300 mg, polyvinyl alcohol 100 mg, dilute to the required volume with water for injection.

[0024] The emulsified solvent diffusion method was used for preparation: the prescribed amount of rhein, hesperidin and PLGA were accurately weighed, dissolved in an appropriate amount of methanol, and dissolved by ultrasonication in a water bath to serve as the organic phase; the prescribed amount of poloxamer 188 (P188) and polyvinyl alcohol (PVA) were dissolved in pure water to serve as the aqueous phase; the organic phase was added to the aqueous phase, ultrasonically emulsified in a water bath for 10 minutes, and the organic solvent was removed by rotary evaporation under reduced pressure at 33°C to obtain rhein and hesperidin co-loaded nanoparticles.

[0025] Example 3: Preparation of rhein and hesperidin co-loaded nanoparticles The preferred ratio of rhein and hesperidin co-loaded nanoparticles in this embodiment is (rhein: hesperidin = 5:1) Rhein 5 mg, hesperidin 1 mg, PLGA 50 mg, poloxamer 188 300 mg, polyvinyl alcohol 100 mg, dilute to the required volume with water for injection.

[0026] The emulsified solvent diffusion method was used for preparation: the prescribed amount of rhein, hesperidin and PLGA were accurately weighed, dissolved in an appropriate amount of methanol, and dissolved by ultrasonication in a water bath to serve as the organic phase; the prescribed amount of poloxamer 188 (P188) and polyvinyl alcohol (PVA) were dissolved in pure water to serve as the aqueous phase; the organic phase was added to the aqueous phase, ultrasonically emulsified in a water bath for 10 minutes, and the organic solvent was removed by rotary evaporation under reduced pressure at 33°C to obtain rhein and hesperidin co-loaded nanoparticles.

[0027] The particle size detection results of the nanoparticles co-loaded with rhein and hesperidin in different ratios in the above Examples 1-3 are similar. The average particle size of the nanoparticles with a ratio of rhein to hesperidin of 1:1 is (114.1 ± 0.98) nm, the average particle size of the nanoparticles with a ratio of rhein to hesperidin of 1:5 is (132.5 ± 0.10) nm, and the average particle size of the nanoparticles with a ratio of rhein to hesperidin of 5:1 is (127.7 ± 0.16) nm. Preferably, the ratio of rhein to hesperidin is 1:1 for subsequent experiments.

[0028] Experimental Example 1: Characterization of Rhein and Hesperidin Co-loaded Nanoparticles The prepared sample was diluted to a certain extent, and its average particle size and Zeta potential were measured by a laser particle size analyzer. The sample was dropped on a copper mesh, negatively stained with 2% (w / v) phosphotungstic acid, and the morphology and particle size of the nanoemulsion were observed by transmission electron microscopy (TEM).

[0029] Test results such as Figure 1 and Figure 2 As shown: After 2% phosphotungstic acid negative staining, the electron microscopy image shows that rhein and hesperidin co-loaded nanoparticles ( Figure 1 ) surface is intact, round, and the particle size distribution is uniform, and the particle size is about 100nm. The rhein and hesperidin co-loaded nanoparticles ( Figure 2 ) can be seen from the particle size distribution of the nanoemulsion that the particle size distribution is uniform, the average particle size is about (114.1 ± 0.98) nm, the polydispersity index (PDI) is 0.15 ± 0.01, and the Zeta potential is (5.11 ± 0.89) mV.

[0030] Experimental Example 2: Stability Study of Nanoparticles Co-loaded with Rhein and Hesperidin The effect of high-speed centrifugation on the stability of rhein and hesperidin co-loaded nanoparticles was investigated. After high-speed centrifugation, the nanoparticles showed no drug precipitation, flocculation, or stratification compared with the untreated nanoparticles, and there was no significant difference in particle size and PDI ( Figure 3 A), indicating that the stability of rhein and hesperidin co-loaded nanoparticles was not affected by high-speed centrifugation.

[0031] The effect of heating and cooling cycles on the stability of rhein and hesperidin co-loaded nanoparticles was investigated. After the heat-cooling cycle test, the particle size and PDI of the nanoemulsion had no significant difference compared with the untreated nanoemulsion ( Figure 3 B), Rhein and hesperidin co-loaded nanoparticles have good stability at 4°C and 25°C.

[0032] Experimental Example 3: Study on the in vitro release of rhein and hesperidin co-loaded nanoparticles To investigate the in vitro release of rhein and hesperidin co-loaded nanoparticles, 1 mL of rhein and hesperidin co-loaded nanoparticles, rhein and hesperidin APIs were precisely drawn, and three copies were prepared in parallel and placed in pre-treated dialysis bags, and the two ends of the dialysis bags were tied with cotton thread. They were placed in 30 mL of release medium of pH 7.4 phosphate buffer (containing 0.2% Tween-80), and the dialysis bags were completely immersed under the liquid surface. The dialysis bags were oscillated at 37°C (100r / min), and 1 mL of dialysate was drawn from the centrifuge tube at 0.1, 0.5, 0.75, 1, 2, 6, 8, 12, 24, 36, and 48, respectively, and the release medium of the same temperature and pH was supplemented. After the samples were filtered through a 0.45μm microporous filter membrane, the contents of rhein and hesperidin were determined by UV spectrophotometry, and the cumulative release curve was drawn ( Figure 4 The results showed that the cumulative release of the raw material was greater than 90% within 6 hours, while the cumulative release of rhein and hesperidin co-loaded nanoparticles reached 90% within 24 hours, indicating that rhein and hesperidin co-loaded nanoparticles can slow down the release rate of rhein and hesperidin and achieve a sustained release effect.

[0033] Experimental Example 4: Antitumor effect of rhein and hesperidin co-loaded nanoparticles Mouse 4T1 breast cancer cells in the logarithmic growth phase were digested with 0.25% trypsin and centrifuged at 1000 rpm for 3 minutes to adjust the cells to an appropriate concentration. The cells were placed on ice for later use. After mixing the cells, 7×10 5 cells were implanted in the first pair of mammary fat pads under the left armpit of mice ( Figure 5 A).

[0034] The mice were divided into five groups: PBS group, blank nanoparticle group (Blank-NP), rhein nanoparticle group (Rhe-NP), hesperidin nanoparticle group (Hes-NP), and rhein and hesperidin co-loaded nanoparticle group (HR-NP), with 5-6 mice in each group. The drugs were intraperitoneally injected every day, and the tumor diameter was measured every two days. The weight changes of mice during the drug administration period were recorded ( Figure 5 B), calculate the tumor volume ( Figure 5C) Using mouse in vivo fluorescence technology to detect changes in tumor luminescence intensity ( Figure 5 F), Calculation of BLI of different drug treatment groups at different times rel Ratio curve ( Figure 5 G). Tumor-bearing mice were killed on the 14th day of administration, and the tumors were dissected out after dissection ( Figure 5 E) Record the tumor weight of tumor-bearing mice ( Figure 5 D), H&E stained tumor tissue ( Figure 5 H).

[0035] The experimental results showed that compared with PBS and blank nanoparticles, nanoparticles co-loaded with rhein and hesperidin can significantly reduce the growth of breast cancer, and the combined use of rhein and hesperidin has stronger anti-tumor activity than single components. In summary, nanoparticles co-loaded with rhein and hesperidin can also significantly reduce the growth of tumors in cancer cells such as melanoma, gastric cancer, lung cancer, pancreatic cancer, liver cancer, kidney cancer, cervical cancer, uterine cancer, skin cancer, prostate cancer, thyroid cancer, leukemia, lymphoma, esophageal cancer, oral cancer, intestinal cancer, nasal cancer, head and neck cancer, etc.

[0036] Experimental Example 5: Analysis of Immune Cells in Tumor Tissue Regulatory T cells (Tregs) are a type of T lymphocyte with negative immune regulatory function. Myeloid-derived suppressor cells (MDSCs) are derived from bone marrow progenitor cells and immature myeloid cells and are a group of cells with strong immunosuppressive function. Tumor-associated macrophages (TAMs) are the most abundant immune cells in tumor tissues and can participate in natural immunity and acquired immunity to respond to the tumor microenvironment. TAMs can be activated into the classic M1 type and the alternative M2 type, which are mainly involved in intracellular killing of pathogens and anti-tumor immune responses. The M2 type is more inclined to immunoregulation and tumor promotion activity.

[0037] About 100 mg of tumor tissue was collected and placed in a centrifuge tube containing PBS. The tumor was cut into small pieces, digestive enzyme solution was added, single cells were separated by grinding, centrifugation was performed, cells were collected, red blood cell lysis solution was added, red blood cells were lysed, PBS was washed twice, centrifuged, and single cell suspension was obtained after re-suspending. CD8+ T cells in tumor tissue were analyzed by flow cytometry ( Figure 5 A, B), CD8+T cells in lymphoid tissue ( Figure 5 C, D), CCL2 in tumor tissue ( Figure 5 E) M1 / M2 ratio ( Figure 5 F) Tregs Figure 5 G), MDSCs ( Figure 5 H) situations, such as Figure 6 shown.

[0038] The experimental results showed that compared with the control group, the CD8+T cells and M1 / M2 ratio were significantly upregulated, while Tregs, MDSCs and CCL2 were significantly downregulated after the combined use of rhein and hesperidin, indicating that the combined use of rhein and hesperidin can regulate the tumor microenvironment and inhibit tumor growth.

[0039] Experimental Example 6: Immunofluorescence detection of α-SMA and CD8 in tumor tissue + Changes in T expression The tumor microenvironment of triple-negative breast cancer is composed of tumor cells, cancer-associated fibroblasts, adipocytes, infiltrating immune cells, stromal cells and other cell types, as well as non-cellular tissue components. Cancer-associated fibroblasts, as important stromal cells in the tumor microenvironment, mediate endothelial hyperplasia reactions and are the main driving factor of the immunosuppressive microenvironment that promotes tumor growth, thereby reducing the effectiveness of anti-tumor treatment.

[0040] After the treatment, the tumor-bearing mice in each group were killed by cervical dislocation, and the tumor tissues were removed and fixed in 4% paraformaldehyde. Part of the tumor tissue was dehydrated, embedded in paraffin, and sliced. The slices were placed in a 37°C oven for 2 h until dried, and then dewaxed. After heating and repairing with sodium citrate repair solution (pH 6.0), PBS was used for washing, and the primary antibody (CD8, α-SMA antibody) was incubated at 4°C overnight. After washing with PBS the next day, the secondary antibody was incubated at 37°C for 30 min. After washing with PBS, the slices were sealed with anti-fluorescence quenching sealing agent, and the digital pathology scanner VS120-S6-W (OLYMPUS, Japan) was used to observe the staining results of the slices. Figure 7 The results showed that compared with the control group, the combined use of rhein and hesperidin reduced α-SMA in the tumor site and increased CD8+T cell infiltration.

[0041] Experimental Example 7: H&E staining analysis of the main organs and tumor tissues of tumor-bearing mice and blood biochemical indicators After administration, blood was collected from the eyeballs of tumor-bearing mice. 1.5 mL of blood was placed in an EP tube and placed on ice for 30 min before centrifugation (3500 rpm, 10 min, 4°C) to obtain the supernatant for the detection of liver and kidney function indicators. The results are shown in the figure ( Figure 8 A) After administration, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (CREA) and blood urea nitrogen (BUN) in tumor-bearing mice did not change significantly compared with those in the PBS group. The results showed that there was no obvious toxicity to the liver and kidneys in each administration group.

[0042] After the administration, the organs (heart, liver, spleen, lung, kidney) and tumor tissues of tumor-bearing mice were obtained and fixed in 4% paraformaldehyde, and immediately prepared into paraffin sections. The paraffin sections were dewaxed, stained with hematoxylin for 5 minutes, differentiated with hydrochloric acid ethanol for 5 seconds, stained with eosin for 5 minutes, and then dehydrated with ethanol and xylene gradients, and finally sealed with neutral gum. The sections were placed under a 200x microscope for observation and photography ( Figure 8 B). The cell morphology of the main organs (heart, liver, spleen, lung, and kidney) in each drug treatment group was relatively complete, the nuclei did not increase significantly, and no significant damage was observed.

[0043] The above experiments show that the combined use of rhein and hesperidin has a good anti-tumor effect.

Claims

1. A combined Chinese medicine composition for treating tumors by improving the tumor immune microenvironment, characterized in that: The traditional Chinese medicine composition comprises rhein and hesperidin.

2. A combined Chinese medicine composition for treating tumors by improving the tumor immune microenvironment as claimed in claim 1, characterized in that: Rhein is rhein monomer or a pharmaceutically acceptable salt thereof, and hesperidin is hesperidin monomer or a pharmaceutically acceptable salt thereof.

3. A combined Chinese medicine composition for treating tumors by improving the tumor immune microenvironment as claimed in claim 1, characterized in that: The mass ratio of rhein to hesperidin is 1-10:1-10, and preferably the mass ratio of rhein to hesperidin is 1:

1.

4. A combined Chinese medicine composition for treating tumors by improving the tumor immune microenvironment as claimed in claim 1, characterized in that: The dosage form of the combined Chinese medicine composition is any pharmaceutically acceptable dosage form, preferably nanoparticles, liposomes, nanoemulsions, microcapsules, microspheres, micropellets, solid dispersions, and more preferably nanoparticles.

5. The combined Chinese medicine composition for treating tumors by improving the tumor immune microenvironment according to claim 1, characterized in that: The traditional Chinese medicine composition also includes any one or more combinations of pharmaceutical excipients acceptable in pharmaceutical science.

6. A combined Chinese medicine composition for treating tumors by improving the tumor immune microenvironment as claimed in claim 5, characterized in that: Pharmaceutical excipients include polylactic acid-glycolic acid copolymer, poloxamer 188 and polyvinyl alcohol.

7. The combined Chinese medicine composition for treating tumors by improving the tumor immune microenvironment according to claim 1, characterized in that: The administration form of the Chinese medicine composition includes any one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration or transdermal administration, preferably intraperitoneal injection.

8. Use of the combined Chinese medicine composition according to any one of claims 1 to 7 in the preparation of anti-tumor drugs, wherein the tumors include melanoma, gastric cancer, lung cancer, pancreatic cancer, liver cancer, kidney cancer, breast cancer, cervical cancer, uterine cancer, skin cancer, prostate cancer, thyroid cancer, leukemia, lymphoma, esophageal cancer, oral cancer, intestinal cancer, nasal cancer, head and neck cancer, preferably breast cancer.

9. Use of the combined Chinese medicine composition according to any one of claims 1 to 7 in the preparation of a drug for improving the tumor immune microenvironment.

10. The use according to claim 9, characterized in that The immune microenvironment includes cancer-associated fibroblasts, cancer-associated adipocytes, tumor-associated macrophages, tumor-associated neutrophils, and tumor-infiltrating lymphocytes, preferably cancer-associated fibroblasts and cancer-associated adipocytes.

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