CN116036281A

CN116036281A - Application of sodium-glucose cotransporter 2 inhibitor in preparation of tumor immunotherapy medicaments

Info

Publication number: CN116036281A
Application number: CN202211707300.4A
Authority: CN
Inventors: 侯嘉杰; 张海燕
Original assignee: Sun Yat Sen University Cancer Center
Current assignee: Sun Yat Sen University Cancer Center
Priority date: 2022-12-27
Filing date: 2022-12-27
Publication date: 2023-05-02

Abstract

The invention provides application of a sodium-glucose cotransporter 2 inhibitor in preparing tumor immunotherapy medicaments. According to the research result of the invention, the sodium-glucose cotransporter 2 inhibitor can regulate and control the immune microenvironment in liver tissues where tumors occur, can reduce the number of immature dendritic cells in the liver tissues, reduce the inhibition effect of the immature dendritic cells on a local immune system, and recover the killing function of T lymphocytes on the tumors, thereby achieving the purpose of inhibiting the growth of the tumors, and having remarkable treatment effect on the liver tumors, so the sodium-glucose cotransporter 2 inhibitor can be used for preparing tumor immunotherapy medicaments or realizing synergistic effect in combination with tumor immunotherapy. In addition, the SGLT2 inhibitor has good safety and convenient administration, and has liver tissue specific anticancer value of the SGLT2 inhibitor and excellent application prospect in preparing tumor therapeutic drugs, especially tumor immunotherapy drugs.

Description

Application of sodium-glucose cotransporter 2 inhibitor in preparation of tumor immunotherapy medicaments

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to application of a sodium-glucose cotransporter 2 inhibitor in preparation of tumor immunotherapy medicaments.

Background

Under normal conditions, the human immune system can recognize and clear tumor cells in the tumor microenvironment, but for survival and growth, the tumor cells can adopt different strategies, so that the immune system of the human body is inhibited and can not normally kill the tumor cells, thereby surviving in each stage of anti-tumor immune response. Tumor immunotherapy is a therapeutic method for controlling and eliminating tumors by restarting and maintaining tumor-immune circulation and restoring the normal anti-tumor immune response of the body. Tumor immunotherapy includes monoclonal antibody immune checkpoint inhibitors, therapeutic antibodies, cancer vaccines, cell therapies, small molecule inhibitors, and the like. In recent years, good information on tumor immunotherapy is continuous, and strong anti-tumor activity has been shown in the treatment of various tumors such as melanoma, non-small cell lung cancer, renal cancer, prostate cancer and other solid tumors, and a plurality of tumor immunotherapeutic drugs have been applied in clinical batches. Tumor immunotherapy is evaluated as the most important scientific breakthrough in 2013 by journal of science due to its excellent therapeutic effect and innovation; discoverers of related tumor immunity mechanisms were awarded a 2018 Norbeol medical prize.

The immune therapy is the same as other antitumor drugs, and it is difficult to avoid the occurrence of drug resistance. In the clinical practice of immunotherapy, only about 20% of patients respond to immunotherapy, and some of them relapse and lose the response after a period of treatment. The immunotherapeutic resistance patterns can be classified into primary resistance (primary resistance), adaptive resistance (adaptive immune resistance) and acquired resistance (acquired resistance) according to the mechanism of drug resistance. Such as the liver, is also one of the most common target organs for cancer metastasis (tumors occurring in the liver include primary/metastatic liver tumors) as a key organ involved in the metabolism of the body. With the physiological properties of the 'immune preferential organ', various types of immunosuppressive cells exist in the liver microenvironment in infiltration, and the later cells possibly weaken the killing function of T cells, so that novel immunotherapeutic drugs such as immune checkpoint inhibitors and the like are disabled. A recent study reviews the immune checkpoint inhibitor treatment response of metastatic melanoma and lung cancer patients and finds that liver metastases patients respond less to immunotherapy and have a shorter survival time than patients with other organ metastases—liver metastases can lead to systemic immune tolerance, and even under immune checkpoint inhibitor treatment pressure, systemic multi-site tumor hyperprogression can occur. The primary/metastatic liver tumor responds poorly to chemotherapy, radiotherapy and immunotherapy, and liver-specific immunotherapy resistance also occurs in a new generation of immunotherapeutic drugs represented by immune checkpoint monoclonal antibodies.

The clinical practice of tumor immunotherapy is frustrated, and the dilemma of breaking liver tumor immunotherapy by exploring novel immunotherapeutic drugs is a urgent matter.

Disclosure of Invention

Aiming at the problem of poor tumor immunotherapy effect in the prior art, the invention aims to provide the application of the sodium-glucose cotransporter 2 inhibitor in preparing tumor immunotherapy medicaments and provides a new choice for tumor immunotherapy.

The primary purpose of the invention is to provide an application of the sodium-glucose cotransporter 2 inhibitor in preparing tumor immunotherapy medicaments or preparing tumor immunotherapy synergists.

It is a further object of the present invention to provide the use of sodium-glucose cotransporter 2 inhibitors in combination with a tumor immunotherapeutic agent for the manufacture of a medicament for the treatment of tumors.

It is still another object of the present invention to provide a therapeutic agent for liver tumor.

The invention realizes the aim by the following technical scheme:

the invention discovers the tumor immunology function and mechanism of the sodium-glucose cotransporter 2 (SGLT 2) inhibitor, and shows the important value of the sodium-glucose cotransporter 2 (SGLT 2) inhibitor in the liver immune microenvironment remodeling and primary/metastatic liver tumor immunotherapy for the first time. The SGLT2 inhibitor has a treatment mechanism of primary/metastatic liver tumor different from other hypoglycemic drugs, has better treatment effect than certain immunotherapeutic drugs, and is expected to break the clinical dilemma of primary/metastatic liver tumor immunotherapy.

Specifically, the invention discovers that (1) the SGLT2 inhibitor can not generate obvious curative effect on subcutaneous tumors of mice, but can effectively inhibit the growth of primary/metastatic liver tumors, and the tumor treatment effect of the SGLT2 inhibitor has organ specificity, but does not have a floods inhibition effect. (2) SGLT2 inhibitors show obvious curative effects on various primary/metastatic liver tumors. (3) The SGLT2 inhibitor can treat tumor-bearing mice with complete immune systems, and has no obvious curative effect on liver tumors of combined immunodeficiency mice; from this, it can be determined that SGLT2 inhibitors exert therapeutic effects by modulating the liver immune microenvironment, rather than acting directly on tumor cells. (4) SGLT2 inhibitor treatment can effectively reduce immature dendritic cells which play a role in immunosuppression in liver, thereby restoring the killing function of T lymphocytes on tumors.

The liver immune microenvironment can be regulated and controlled based on the SGLT2 inhibitor, so that the SGLT2 inhibitor can improve the effect of tumor immunotherapy and can be used as a novel tumor immunotherapy drug or an immunotherapy synergist; meanwhile, the SGLT2 inhibitor and the tumor immunotherapy drug can obviously improve the treatment effect on liver tumors, and provide more choices for tumor treatment.

Therefore, the following technical schemes are all within the protection scope of the invention:

the invention provides an application of a sodium-glucose cotransporter 2 inhibitor in preparing tumor immunotherapy medicaments or preparing tumor immunotherapy synergists.

Preferably, the drug is a drug capable of remodelling the tissue immune microenvironment.

Preferably, the tumor immunotherapy comprises an immune checkpoint inhibitor therapy.

The invention also provides application of the sodium-glucose cotransporter 2 inhibitor in combination with a tumor immunotherapy drug in preparing a drug for treating tumors.

Preferably, the tumor immunotherapy drug comprises an immune checkpoint inhibitor therapy drug.

More preferably, the immune checkpoint inhibitor therapy drug is a PD-1 antibody.

Preferably, the tumor is one that is resistant to immunotherapy.

Preferably, the tumor is a liver tumor, including a primary liver tumor, a metastatic liver tumor.

Preferably, the metastatic liver tumor comprises a colon cancer liver metastasis, a breast cancer liver metastasis or a melanoma liver metastasis.

The invention also provides a tumor immunotherapy medicament comprising a sodium-glucose cotransporter 2 inhibitor, preferably further comprising a tumor immunotherapy medicament.

More preferably, the tumor immunotherapy drug comprises an immune checkpoint inhibitor therapy drug.

Preferably, the composition further comprises pharmaceutically acceptable auxiliary materials.

Sodium-glucose cotransporter 2 inhibitors of the invention include, but are not limited to, irinotecan, canagliflozin.

The mouse model experiment proves that the daily oral SGLT2 inhibitor can effectively treat primary/metastatic liver tumors, and has no treatment effect on the same tumor cells at other parts, thus indicating the specific curative effect of the sodium-glucose cotransporter 2 (SGLT 2) inhibitor on the primary/metastatic liver tumors.

Experimental data indicate that the immunobiological basis of the SGLT2 inhibitor is to exert curative effect by regulating the liver immune microenvironment, and not directly act on tumor cells.

The technical scheme of the invention has the following beneficial effects:

the invention discovers that the sodium-glucose cotransporter 2 inhibitor can regulate and control the immune microenvironment of liver tumor, reduce the number of immature dendritic cells in liver, reduce the inhibition effect of the immature dendritic cells on the immune system in liver, and recover the killing function of T lymphocytes on tumor, thereby achieving the purpose of inhibiting the growth of liver tumor and having remarkable treatment effect on liver tumor. The SGLT2 inhibitor has good safety and convenient administration, and has excellent transformation prospect for treating liver tumors.

Drawings

Fig. 1 shows the therapeutic effect of the SGLT2 inhibitor ASP1941 of example 1 on MC38 subcutaneous tumors.

FIG. 2 is a graph showing the therapeutic effect of SGLT2 inhibitor ASP1941 of example 2 on primary/metastatic liver tumors; wherein, fig. 2A shows the therapeutic effect of SGLT2 inhibitor ASP1941 on colorectal cancer MC38 liver metastasis; FIG. 2B is a graph showing the therapeutic effect of SGLT2 inhibitor ASP1941 on liver metastasis of breast cancer cells E0771; FIG. 2C is a graph showing the therapeutic effect of SGLT2 inhibitor ASP1941 on melanoma cell B16F10 liver metastases; fig. 2D shows the therapeutic effect of SGLT2 inhibitor ASP1941 on hepatoma cell Hepa1-6 hepatoma in situ.

Fig. 3 is a graph showing the therapeutic effect of SGLT2 inhibitor ASP1941 on liver metastasis in immunodeficient mice of example 3.

FIG. 4 is a spectral flow analysis of example 4; wherein, fig. 4A is an analysis result of liver; fig. 4B shows the peripheral blood analysis results.

FIG. 5 is an experimental result of example 5 immature dendritic cells co-cultured in vitro with activated CD8T cells.

FIG. 6 is a graph showing the change in weight of colon cancer MC38 liver metastases following administration of the SGLT2 inhibitor ASP1941 in combination with anti-PD-1 of example 6.

Detailed Description

The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.

Reagents and materials used in the following examples are commercially available unless otherwise specified.

EXAMPLE 1SGLT2 inhibitors have no therapeutic effect on colorectal cancer cell (MC 38) subcutaneous tumors

1. Experimental method

Wild type C57BL/6 mice (SPF-class animal house raised in Experimental animal center at North school of China university) were perfused daily with 1mg/kg of SGLT2 inhibitor ASP1941 (Igliflozin, selleck, cat# S8637) or control solvent (physiological saline).

7 days after administration, colon cancer MC38 cells (from the cell bank of the unit) are injected into the wild type C57BL/6 mice, and a wild type mouse colon cancer MC38 cell subcutaneous injection tumor-bearing model is constructed, wherein the specific process is as follows:

taking MC38 cells of colon cancer in logarithmic growth phase, digesting with 0.25% trypsin, centrifuging to remove supernatant, centrifuging with serum-free culture solution for 2 times, calculating cell number, adjusting cell concentration, and concentrating cell suspension in PBS to obtain cell concentration of 1.0X10 × ⁷ Colon cancer MC38 cells (0.1 mL/mouse) were injected subcutaneously once per mL in the left flank of C57BL/6 mice. The successful subcutaneous tumor model is single tumor, without double tumor, ectopic tumor and abnormal tumor, and gradually increases the volume, and the volume can reach 1500mm about 21-28 days ³ 。

The SGLT2 inhibitor ASP1941 or physiological saline was continued daily after tumor inoculation, and the specific dose was the same as before tumor inoculation. Tumor volume was measured every 2-3 days from day 5 after tumor inoculation, tumor major and minor diameters were measured 2 times, according to formula v=1/2 ab ² Calculating tumor volume and drawing tumor growth curve. Mice were sacrificed 4 weeks after tumor loading, tumor growth curves were drawn and tumor samples were collected for evaluation of the effect of drug treatment.

2. Experimental results

The experimental results of the therapeutic effect of the SGLT2 inhibitor ASP1941 on colorectal cancer MC38 cell subcutaneous tumor are shown in fig. 1, and it can be seen that the tumor volume of the mice in the SGLT2 inhibitor ASP1941 group is not significantly different from the tumor volume of the mice in the control group along with the time. The results indicate that the SGLT2 inhibitor ASP1941 has no therapeutic effect on MC38 subcutaneous tumors, and that MC38 subcutaneous tumors have no therapeutic response to SGLT2 inhibitors.

EXAMPLE 2SGLT2 inhibitors specifically treat primary/metastatic liver tumors

1. Experiment of therapeutic Effect of SGLT2 inhibitor ASP1941 on colorectal cancer MC38 liver metastasis

1. Experimental method

6-8 week-old C57BL/6 female wild-type mice were treated daily with 1mg/kg of the gastric lavage SGLT2 inhibitor ASP1941 or control solvent (normal saline).

7 days after administration, a model of liver metastasis by intraspleen injection of wild-type mouse colon cancer MC38 cells was established. The specific process is as follows:

under SPF condition, the C57BL/6 female wild type mice are anesthetized by intraperitoneal injection of 1% pentobarbital, prepared into skin, and the skin is sterilized by right lateral recumbent position. The left subcostal incision was made and an incision parallel to the rib arch, approximately 1cm long, was made and the spleen gently pulled out. Surrounding spleen tissue was isolated, exposing the entire spleen and splenomegaly vessels. A slipknot is reserved at the splenic electrode by 5-0 silk threads, and is not fastened once. Colon cancer MC38 cell suspension (1×10) was aspirated with a 1mL syringe with 27G needle ⁶ ) The needle was inserted approximately 0.5cm from the lower pole of the spleen, taking care not to puncture the spleen. Slow injection of 0.1mL of the cell suspension, the spleen swelling was seen to whiten. Slowly withdraw the needle, quickly tighten the wire reserved in advance to ligate the needle, and prevent the cell suspension from leaking out of the needle hole. The spleen was gently squeezed with a wet cotton swab for about 1 minute to shrink the spleen and restore color. Ligating splenomegaly vessels, and resecting spleen after isolating perispleen tissue, taking care not to damage pancreatic tissue on the back of the spleen.

The SGLT2 inhibitor ASP1941 or physiological saline was continuously administered daily after the model was established, and the specific administration dose was the same as before the model was established. Mice were observed daily for signs, sacrificed 2 weeks after tumor loading, weighed for open-abdomen, compared for differences in tumor growth between groups, and peripheral blood and tumor tissue specimens were collected (for immunocyte detection in example 4).

2. Experimental results

The weight change of colon cancer MC38 liver metastases following administration of SGLT2 inhibitor ASP1941 is shown in fig. 2A, which shows that the SGLT2 inhibitor ASP1941 group had significantly reduced tumor mass compared to the control group. The results show that the SGLT2 inhibitor ASP1941 has a remarkable treatment effect on MC38 liver metastasis, and the MC38 liver metastasis can generate a remarkable treatment response on the SGLT2 inhibitor.

2. Experiment of therapeutic Effect of SGLT2 inhibitor ASP1941 on breast cancer cell E0771 liver metastasis

1. Experimental method

The specific experimental process is the same as the experimental of the SGLT2 inhibitor ASP1941 on the treatment effect of colon cancer MC38 liver metastasis, and is different in that the experimental process uses breast cancer cells E0771 to replace colon cancer MC38 cells to construct a wild mouse breast cancer cells E0771 spleen internal injection liver metastasis model.

2. Experimental results

The weight change of breast cancer cells E0771 liver metastases after administration of SGLT2 inhibitor ASP1941 is shown in fig. 2B, which shows that the SGLT2 inhibitor ASP1941 group has significantly reduced tumor mass compared to the control group. The result shows that the SGLT2 inhibitor ASP1941 has obvious treatment effect on the breast cancer cell E0771 liver metastasis, and the breast cancer cell E0771 liver metastasis can generate obvious treatment response to the SGLT2 inhibitor.

3. Experimental effect of SGLT2 inhibitor ASP1941 on treatment of melanoma cell B16F10 liver metastasis

1. Experimental method

The specific experimental process is the same as the experimental of the SGLT2 inhibitor ASP1941 on the treatment effect of colon cancer MC38 liver metastasis, and is different in that the experimental process uses melanoma cells B16F10 to replace colon cancer MC38 cells to construct a wild mouse melanoma cells B16F10 spleen internal injection liver metastasis model.

2. Experimental results

Changes in weight of melanoma cell B16F10 liver metastases following administration of SGLT2 inhibitor ASP1941 as shown in fig. 2C, it can be seen that the tumor mass was significantly reduced in the SGLT2 inhibitor ASP1941 group compared to the control group. The result shows that the SGLT2 inhibitor ASP1941 has obvious treatment effect on the liver metastasis of the melanoma cell B16F10, and the liver metastasis of the melanoma cell B16F10 can generate obvious treatment response to the SGLT2 inhibitor.

4. Therapeutic action experiment of SGLT2 inhibitor ASP1941 on hepatoma cell Hepa1-6 hepatoma in situ

1. Experimental method

C57BL/6 male wild-type mice of 6-8 weeks old were treated daily with 1mg/kg of a gastric lavage SGLT2 inhibitor ASP1941 or a control solvent (physiological saline).

After 7 days of administration, a wild mouse hepatoma cell Hepa1-6 hepatoma in situ model was established. The specific process is as follows:

under SPF condition, the mice are anesthetized by intraperitoneal injection of 1% pentobarbital, prepared into skin, and the right lateral recumbent position is taken to disinfect the skin. A midline incision was made to expose the liver. Mouse liver cancer cell line Hepa1-6 cells resuspended in matrigel (1:1 mix) were slowly injected under the liver capsule (1×10) using an insulin syringe ⁶ ) Care was taken to prevent tumor cell leakage. The SGLT2 inhibitor ASP1941 or physiological saline was continuously administered daily after the tumor model was established, and the specific administration dose was the same as before tumor inoculation. Tumor-bearing mice were sacrificed 14 days after inoculation, livers were removed and weighed, and differences in tumor growth between groups were compared.

2. Experimental results

The weight change of hepatoma cells Hepa1-6 liver in situ tumor after administration of SGLT2 inhibitor ASP1941 is shown in fig. 2D, and it can be seen that the SGLT2 inhibitor ASP1941 group has significantly reduced tumor quality compared with the control group. The result shows that the SGLT2 inhibitor ASP1941 has obvious treatment effect on hepatoma cell Hepa1-6 hepatoma in situ, and the hepatoma cell Hepa1-6 hepatoma can generate obvious treatment response to the SGLT2 inhibitor.

Example 3 intrahepatic immune cells are important factors in mediating the tumor therapeutic effects of SGLT2 inhibitors

1. Experimental method

Severe immunodeficiency (NOG) mice were treated daily with 1mg/kg dose of the gastric lavage SGLT2 inhibitor ASP1941 or control solvent (normal saline).

7 days after administration, a model of the immune deficiency of colon cancer MC38 cell by intrasplenic injection of liver metastasis was established, and the specific procedure is the same as in example 2.

The SGLT2 inhibitor ASP1941 or physiological saline was continuously administered daily after the tumor model was established, and the specific administration dose was the same as before tumor inoculation. Mice were sacrificed 2 weeks after tumor-bearing, livers were weighed, and differences in tumor growth between groups were compared.

2. Experimental results

The change in weight of liver metastases in immunodeficient mice following administration of SGLT2 inhibitor ASP1941 is shown in fig. 3, which shows that SGLT2 inhibitor ASP1941 group has no significant difference in tumor mass from control group and NOG murine liver metastases do not respond effectively to SGLT2 inhibitor treatment. The results indicate that the inhibition of liver metastasis by SGLT2 inhibitors is dependent on immune cells in vivo.

Example 4SGLT2 inhibitors remodelling of the tumor immune microenvironment in the liver

1. Experimental method

(1) Detection of liver immune cells

Spectral flow techniques were used to analyze changes in immune cells in liver of colon cancer MC38 liver metastasis model mice following treatment with SGLT2 inhibitors of example 2. Example 2 model mouse livers were taken after mice were sacrificed and homogenized as a single cell suspension with gentleMACS Dissociator (Miltenyi). After digestion, cells were screened on a 70 μm screen, centrifuged on a 35%/55%/75% percoll density gradient, interface lymphocytes were collected, washed with PBS, resuspended with FACS solution, labeled with streaming antibodies (CD 45, tcrβ, tcrγδ, F4/80, CD11b, ly-6C, ly-6G, NK 1.1.1, CD4, CD8, CD19, CD11c, CD103, CD69, CD44, MHC II) and finally added to streaming cell counting microspheres. The ratio and number of liver infiltrating immune cells were measured using a full spectrum flow cytometer (Cytek Aurora) and the differences between groups were counted.

(2) Detection of peripheral blood immune cells

The changes in peripheral blood immune cells of colon cancer MC38 liver metastasis model mice following treatment with SGLT2 inhibitors of example 2 were analyzed using spectroscopic flow techniques. Example 2 model mice peripheral blood was collected after mice were sacrificed, erythrocytes were lysed using erythrocyte lysate, peripheral blood immune cells were centrifuged, washed with PBS, resuspended with FACS solution, labeled with streaming antibodies (CD 45, tcrp, tcrγδ, F4/80, CD11b, ly-6C, ly-6G, NK 1.1.1, CD4, CD8, CD19, CD11c, CD103, CD69, CD44, MHC II) and finally added to streaming cell count microspheres. The ratio and number of liver infiltrating immune cells were measured using a full spectrum flow cytometer (Cytek Aurora) and the differences between groups were counted.

2. Experimental results

The experimental results are shown in FIG. 4, and it can be seen that the SGLT2 inhibitor group intra-hepatic CD11C ⁺ F4/80 ⁺ The proportion of immature dendritic (imDC) cells was significantly reduced, the T cell proportion was significantly increased (FIG. 4A), and the peripheral blood immature dendritic cellsAnd the T cell ratio was not significantly changed (fig. 4B). Experimental results show that the SGLT2 inhibitor can remodel the immune microenvironment of the tumors in the liver.

EXAMPLE 5 in-liver immature dendritic cells exert immunosuppressive function

1. Experimental method

Flow sorting purification the immature dendritic cells in the liver of the colon cancer MC38 liver metastasis model mouse constructed in example 2 were tested for proliferation of CD8T cells after co-culture with activated CD8T cells in vitro.

The method comprises the following steps: taking a colon cancer MC38 liver metastasis model mouse, extracting liver infiltration immune cells, and separating and purifying CD11C by using a flow cytometry ⁺ F4/80 ⁺ Immature dendritic cells. Cell viability by direct counting with microscope after staining with trypan blue>95%. Another 6-8 week old C57BL/6 wild type mouse was isolated and purified spleen CD8T cells by immunomagnetic beads and labeled with CTV and co-cultured with the purified immature dendritic cells in 96-well plates (2X 10) ⁵ Well, concentration gradient immature dendritic cells: CD8T cells were 1:1, a step of; 1:10;1:100 And stimulated with anti-CD 3/CD28 magnetic beads. A CD3/CD28 control group was set. After 72 hours, CD8T cell proliferation was examined by flow cytometry.

2. Experimental results

As shown in FIG. 5, it can be seen that the immature dendritic cells can effectively inhibit proliferation of CD8T cells, and exhibit immunosuppressive function.

Example 6SGLT2 inhibitors enhance the therapeutic response of metastatic liver tumors to immune checkpoint blockers

1. Therapeutic action experiment of SGLT2 inhibitor ASP1941 combined immune checkpoint blocker anti-PD-1 on MC38 liver metastasis

1. Experimental method

6-8 week old C57BL/6 female wild-type mice were treated with the gastric SGLT2 inhibitor ASP1941 or control solvent (normal saline) at a dose of 1mg/kg daily for 21 consecutive days.

7 days after administration, a model of liver metastasis by intraspleen injection of wild-type mouse colon cancer MC38 cells was established. The specific procedure was as in example 2. After the model is built, the SGLT2 inhibitor ASP1941 or normal saline is continuously administered, and the specific dosage is the same as before the model is built. 250 μg of anti-PD-1 blocking antibody (BioXcell, BP 0146) was subcutaneously injected 6 days after tumor-bearing, one needle every 3 days, followed by 3 needles. Mice were sacrificed 2 weeks after tumor-bearing, and the difference in tumor growth between groups was compared.

2. Experimental results

The weight change of colon cancer MC38 liver metastasis after combined administration (SGLT 2 inhibitor ASP1941+anti-PD-1) is shown in FIG. 6, and it can be seen that the tumor quality of the anti-PD-1 single-drug treatment group is not significantly changed compared with that of the control group. Liver metastatic mice developed immune therapy resistance. Notably, the combined administration (SGLT 2 inhibitor ASP 1941+anti-PD-1) group had significantly reduced tumor mass compared to the control group. The result shows that the SGLT2 inhibitor ASP1941 combined with the immune checkpoint blocker anti-PD-1 has a remarkable treatment effect on MC38 liver metastasis, and the SGLT2 inhibitor can effectively enhance the treatment response of metastatic liver tumors to the immune checkpoint blocker.

The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.

Claims

1. The application of sodium-glucose cotransporter 2 inhibitor in preparing tumor immunotherapy medicine or tumor immunotherapy synergist.

2. The use according to claim 1, wherein the medicament is a medicament capable of remodelling the tissue immune microenvironment and/or enhancing the tumor immune response.

3. The use of claim 1, wherein the tumor immunotherapy comprises an immune checkpoint inhibitor therapy.

4. The application of sodium-glucose cotransporter 2 inhibitor in preparing medicine for treating tumor.

5. The use of claim 4, wherein the tumor immunotherapeutic agent comprises an immune checkpoint inhibitor therapy agent.

6. The use according to claims 1-5, wherein the tumor is a tumor that is resistant to immunotherapy.

7. The use according to claims 1-5, wherein the tumor comprises a primary liver tumor, a metastatic liver tumor.

8. The use according to claim 7, wherein the metastatic liver tumor comprises colon cancer liver metastasis, breast cancer liver metastasis or melanoma liver metastasis.

9. A tumor immunotherapeutic agent comprising a sodium-glucose cotransporter 2 inhibitor, preferably a tumor immunotherapeutic agent.

10. The medicament of claim 9, further comprising a pharmaceutically acceptable excipient.