Compositions and methods for combination treatment of cancer

Abstract

The present invention relates to combination therapy compositions and methods of combination therapy for the treatment of cancer. The present invention discloses a composition comprising: (i) an effective amount of a programmed cell death protein-1 (PD-1) inhibitor capable of blocking a PD-1 pathway, partially recovering the function of T cells, and enabling these cells to continue killing tumor cells; and (ii) an effective amount of a glucosylceramide synthetase (GCS) inhibitor capable of enhancing the activity of the immune system; wherein the immunosuppressive agent is administered intravenously by using an intravenous dosage form, and the GCS inhibitor is administered orally by using an oral dosage form. The tumor growth can be effectively inhibited by using the composition and the combined treatment method disclosed by the invention.

Description

Technical Field

[0001] The present invention relates to a composition and method for combined therapy for treating cancer, in particular to a composition and method comprising a programmed death 1 protein inhibitor and a glucosylceramide synthase inhibitor, and belongs to the field of medicine. Background Art

[0002] Immunotherapy has become the main effective means to deal with tumors. Activated T cells can effectively eliminate pathogens and abnormal cells, and are the main way of specific immunity of the body. With the in-depth understanding and research of the regulatory mechanism of T cells, people have found that there are immune checkpoint molecules on the surface of T cells, which play a negative regulatory role, protect the body's normal cells from T cell damage, and maintain the body's immune homeostasis, but tumor cells can also use the checkpoint molecules on the surface of T cells to escape T cell elimination. Programmed death receptor-1 (PD-1) is obtained from apoptotic T cell hybridomas. It is an important immunosuppressive molecule that can bind to programmed death ligand-1 (PD-L1) highly expressed in tumor cells, so that the PD-1 signaling pathway is continuously activated, which can inhibit T cell function, thereby causing the immune function to be unable to attack tumor cells. Therefore, PD-1 / PD-L1 inhibitors can promote the restoration of T cell activity, thereby treating a variety of tumors.

[0003] PD-1 / PD-L1 immunotherapy has achieved great success in the past decade. It has been found that PD-1 / PD-L1 immunotherapy has good clinical effects on non-small cell lung cancer, melanoma, head and neck tumors, breast cancer, liver cancer and other cancers. However, the disadvantage is that only a part of patients show clinical response, and most patients cannot benefit from PD-1 / PDL-1 immunotherapy. In addition, a large number of treatment responders will develop acquired resistance after the initial response, leading to treatment failure. At present, the mechanism of primary tumor resistance is still unclear. Insufficient tumor immunogenicity, bare lymphocyte syndrome (MHC dysfunction), irreversible T cell exhaustion, primary resistance to IFN-γ signaling and immunosuppressive microenvironment may cause patients to develop resistance. Therefore, the combination of PD-1 / PD-L1 inhibitors with other therapies is being widely studied (Lei Q, et al., Front Cell Dev Biol. 2020. Resistance Mechanisms of Anti-PD-1PD-L1 Therapy in Solid Tumors).

[0004] UDP-glucose ceramide glycosyltransferase (UGCG) is a gene encoding glucosylceramide synthase (GCS), a unique protein in cells and a key enzyme in the metabolism of glycosphingolipids (GSLs). It is essential for normal cell function. The precursor of all gangliosides is glucosylceramide produced by UGCG. Increased UGCG activity leads to increased levels of gangliosides and / or cerebrosides. UGCG is also associated with multidrug resistance protein 1 (MDR1) gene expression (Marthe-SusannaWegner, et al., Wegner et al. BMC Cancer (2018) 18:153, The UDP-glucoseceramide glycosyltransferase (UGCG) and the link to multidrugresistance protein 1 (MDR1)). GSLs are important components of GSL-rich microdomains in cell membranes and play an important role in molecular signaling, cell crosstalk, and cell adhesion. Tumors often express high levels of GSLs, which lead to high concentrations of free GSLs in the tumor microenvironment through active or passive processes, which interfere with the killing ability of the immune system and make tumors resistant. In addition to drug resistance, high expression of GSLs in tumors can also have a negative impact on the function of T cells and dendritic cells (DCs). Therefore, inhibiting the production of GSLs may be beneficial for cancer immunotherapy, that is, inhibiting UGCG or its encoded GCS may have a positive effect on tumor immunotherapy (TaoZhang, et al., Front Immunol. 2019Jan 29;10:90.,The Role of Glycosphingolipids in Immune Cell Functions).

[0005] Miglustat (IC 50 =20.4μM) and Eliglustat (IC 50=10ng / mL) are both glucosylceramide synthase inhibitors (GCS inhibitors), which were approved by the FDA in 2003 and 2014, respectively, for the treatment of Gaucher disease to reduce the accumulation of glucocerebroside in patients. In some previous mouse tumor models, it has been shown that the use of GCS inhibitors alone may be effective against certain tumors. For example, miglustat can reduce the tumor burden in the mouse Ehrlich ascites tumor model (J Inokuchi. et al., Cancer Lett. 1987 Dec;38(1-2):23-30.,Antitumor activity via inhibition ofglycosphingolipid biosynthesis), and iliglustat may be effective against B cell lymphoma (Elena V Pavlova, et al., J Pathol 2015; 235: 113–124,Inhibition of UDP-glucosylceramide synthase in mice prevents Gaucher disease-associated B-cell malignancy) and myeloma (WO2015166284A1).

[0006] However, in general, when administered alone, tumor therapeutic agents are usually not effective for all patients. Therefore, based on the effects of PD-1 / PD-L1 inhibitors on immunosuppressive sites and the mechanism of action of GSLs on immune cells, attempts can be made to improve tumor cure rates by combining these therapeutic drugs. Summary of the invention

[0007] Based on the effects of GCS inhibitors on immune cells and the current status of insufficient immune response of PD-1 / PD-L1 inhibitors in treating tumors, the present invention uses the GCS inhibitor iliglustat in combination with a PD-1 inhibitor to observe that the combination produces better therapeutic effects on some tumor mouse models, for example, surpassing the effects of monotherapy. The present invention surprisingly found that the combined administration of the GCS inhibitor iliglustat and the PD-1 inhibitor achieved an unexpectedly excellent anti-tumor effect.

[0008] Specifically, the present invention provides a composition for combined cancer treatment, the composition comprising: (i) a therapeutically effective amount of a programmed cell death protein-1 inhibitor (PD-1 inhibitor), and (ii) a therapeutically effective amount of a glucosylceramide synthase inhibitor (GCS inhibitor).

[0009] In some embodiments, the PD-1 inhibitor is selected from: OPDIVO (nivolumab), Keytruda (pembrolizumab), Tuoyi (toripalimab), Daboshu (sintilimab), Erica (camrelizumab), BeiGene (tislelizumab), Aniko (pembrolizumab), and Sepalizumab.

[0010] In some embodiments, the GCS inhibitor is a compound represented by Formula 1, Eliglustat, or a pharmaceutically acceptable salt thereof.

[0011]

[0012] Formula 1 In some embodiments, the salt of the GCS inhibitor is preferably a tartrate salt.

[0013] In some embodiments, the PD-1 inhibitor is administered intravenously using an intravenous dosage form, and the GCS inhibitor is administered orally or intravenously using an oral dosage form or an intravenous dosage form.

[0014] In certain treatment regimens, the combination therapy method is to simultaneously administer therapeutic doses of both of the above-mentioned drugs to an individual in need thereof.

[0015] In some treatment regimens, a PD-1 inhibitor may be administered after a GCS inhibitor.

[0016] In certain treatment regimens, a GCS inhibitor is administered after a PD-1 inhibitor.

[0017] In some embodiments, ilioglustat and pharmaceutically acceptable salts thereof can be administered once or more daily at a therapeutic dose as ilioglustat, and the PD-1 inhibitor can be administered once every 2 weeks, or once every 3 weeks at a therapeutic dose.

[0018] In some embodiments, the duration of treatment with the combination therapy is at least 24 weeks.

[0019] In some embodiments, the cancer can be a solid tumor or a hematological tumor.

[0020] In the above compositions, treatment methods and uses, the individual is a human and the cancer is a solid tumor. In some cases, the solid tumor is non-small cell lung cancer, renal cell carcinoma, head and neck cancer, melanoma, hepatocellular carcinoma, breast cancer, endometrial cancer, ovarian cancer, gastric cancer, colorectal cancer, bladder cancer, pancreatic cancer, glioblastoma, biliary tract cancer, glioma, cervical cancer, advanced or refractory solid tumors, small cell lung cancer, squamous cell carcinoma of the lung, desmoplastic melanoma, mesothelin-positive pleural mesothelioma, esophageal cancer, anal cancer, prostate cancer and thyroid cancer.

[0021] In certain cases, the solid tumor is preferably non-small cell lung cancer or liver cancer.

[0022] In the above-mentioned methods of treatment, compositions and uses, the individual can be a human and the cancer is a blood tumor, in some cases, the blood tumor is acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), diffuse B-cell lymphoma (DLBCL), EBV-positive DLBCL, primary mediastinal large B-cell lymphoma, T cell / histiocyte-rich large B-cell lymphoma, follicular lymphoma, Hodgkin's lymphoma (HL), mantle cell lymphoma (MCL), multiple myeloma (MM), myeloid leukemia-1 protein (Mcl-1), myelodysplastic syndrome (MDS), non-Hodgkin's lymphoma (NHL) or small lymphocytic lymphoma (SLL).

[0023] In certain cases, the hematological tumor is preferably acute lymphoblastic leukemia or multiple myeloma.

[0024] In some of the above methods of treatment, compositions, and uses, the individual is a human, the cancer tests positive for human PD-L1 and is selected from lung cancer, endometrial cancer, urothelial carcinoma, head and neck squamous cell carcinoma, liver cancer, or melanoma.

[0025] The present invention also provides a composition and treatment method for combined cancer treatment, comprising (i) a therapeutically effective amount of a programmed cell death protein-1 inhibitor, and (ii) a therapeutically effective amount of a glucosylceramide synthase inhibitor, wherein the combination therapy exhibits excellent anti-tumor effects compared to the case where they are used alone, and can exhibit anti-tumor effects on various types of cancer.

[0026] In another aspect, the present invention provides a method of treating cancer in an individual, the method comprising simultaneously administering to the individual a combination therapy comprising a PD-1 inhibitor and a GCS inhibitor.

[0027] The combination therapy may also include one or more additional therapeutic agents. The additional therapeutic agent may be, for example, a chemotherapeutic agent, a biological therapeutic agent, an immunogenic agent, etc. other than a GCS inhibitor.

[0028] In some cases, at least one therapeutic agent in the combination therapy is administered using the same dosing regimen (dose, frequency, and duration of treatment) that is typically used when the agent is used as a monotherapy to treat the same cancer. In other cases, the total amount of at least one therapeutic agent received by the patient in the combination therapy is lower than the total amount of the agent when used as a monotherapy, e.g., a smaller dose, a less frequent dose, and / or a shorter duration of treatment.

[0029] The combination therapies disclosed herein can be used before or after surgery, and can be used before, during, or after radiation therapy to remove tumors.

[0030] In some cases, the combination therapy disclosed herein is administered to patients who have not been previously treated with a biologic or chemotherapeutic agent, i.e., treatment-naive patients. In other cases, the combination therapy is administered to patients who have failed to achieve a sustained response after prior treatment with a biologic or chemotherapeutic agent, i.e., treatment-experienced patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is the result of the effect of iliglucostat combined with PD-1 on the viability of A549 cells for 48 hours when Jurkat cells were not activated; Figure 2 This is the result of the effect of iliglucostat combined with PD-1 on A549 cell viability for 48 hours under Jurkat cell activation; Figure 3 This is the result of the effect of combined administration of iliglustat and PD-1 on the tumor volume of C57 lung adenocarcinoma mice; Figure 4 This is the result of the effect of combined administration of iliglustat and PD-1 on the tumor volume of ICR liver cancer mice. DETAILED DESCRIPTION

[0032] Abbreviations and meanings: The terms used herein are only for describing specific embodiments and are not intended to limit the present invention. Unless otherwise specified, all technical terms and scientific terms used herein generally have the same meanings as those generally understood by those skilled in the art to which the present invention belongs.

[0033] The term PD-1 / PD-L1 inhibitor refers to drugs that inhibit tumor growth by blocking the immunosuppressive response mediated by the PD-1 / PD-L1 pathway. The PD-1 receptor expressed by T cells binds to its ligands PD-L1 and PD-L2, which can inhibit T cell proliferation and cytokine production. The PD-1 ligand of some tumor cells is upregulated, and signal transduction through this pathway can inhibit the immune surveillance of activated T cells against tumor cells. The term GCS inhibitor refers to glucosylceramide synthase inhibitors, which is a key enzyme in the synthesis of glucocerebroside. Known GCS inhibitors include iliglustat, meglustat, ambroxol, (D,L) 1-phenyl-2-decanoylamino 3-morpholinol-propanol (PDMP), etc.

[0034] Miglustat: Miglustat is an inhibitor of glucosylceramide synthase, the enzyme that initiates a series of reactions in the synthesis of most glycosphingolipids. Gaucher disease type 1 (GD1) is caused by a functional defect in the enzyme glucocerebrosidase, which mediates the degradation of the glycosphingolipid glucosylceramide. Miglustat acts as a competitive and reversible inhibitor of the enzyme glucosylceramide synthase. Miglustat helps to reduce the rate of glycosphingolipid biosynthesis, thereby reducing the amount of glycosphingolipid substrate to a level that allows the residual activity of the defective glucosylcerabrosidase to be more effective (substrate reduction therapy). In vitro and in vivo studies have shown that Miglustat can reduce the synthesis of glucosylceramide-based glycosphingolipids.

[0035] Eliglustat: Eliglustat capsule is a specific inhibitor of glucosylceramide synthase (IC 50 =10ng / mL), which is a substrate reduction therapy for GD1. In clinical trials, iliglustat capsules have the effects of reducing spleen and liver volume, improving anemia and improving thrombocytopenia.

[0036] In some embodiments, the PD-1 inhibitor is selected from: OPDIVO (nivolumab), Keytruda (pembrolizumab), Tuoyi (toripalimab), Daboshu (sintilimab), Erica (camrelizumab), BeiGene (tislelizumab), and Aniko (pembrolizumab).

[0037] Opdivo (nivolumab): Nivolumab is a human immunoglobulin G4 (IgG4) monoclonal antibody (HuMAb) that binds to the PD-1 receptor, blocks its interaction with PD-L1 and PD-L2, and blocks PD-1 pathway-mediated immunosuppressive responses, including anti-tumor immune responses. In syngeneic mouse tumor models, blocking PD-1 activity can inhibit tumor growth.

[0038] Keytruda (pembrolizumab): Pembrolizumab is a monoclonal antibody that binds to the PD-1 receptor, blocking the interaction between PD-1 and PD-L1 and PD-L2, and relieving the inhibition of immune responses mediated by the PD-1 pathway, including anti-tumor immune responses. In a syngeneic mouse tumor model, blocking PD-1 activity can inhibit tumor growth.

[0039] TuoYi (Teplizumab): Teplizumab can bind to PD-1 on the surface of T cells, blocking its binding to ligands PD-L1 and PD-L2, thereby eliminating the immunosuppression of the PD-1 signaling pathway. Teplizumab can promote T cell proliferation, activate T cell function, and inhibit tumor growth.

[0040] Daboshu (sintilimab): Sintilimab is a human immunoglobulin G4 (IgG4) monoclonal antibody (HuMAb) that binds to the PD-1 receptor, blocks its interaction with PD-L1 and PD-L2, and blocks PD-1 pathway-mediated immunosuppressive responses, including anti-tumor immune responses. In a syngeneic mouse tumor model, blocking PD-1 activity can inhibit tumor growth.

[0041] Erica (carrelizumab): Carrelizumab for injection is a human immunoglobulin G4 (IgG4) monoclonal antibody (HuMAb) that binds to the PD-1 receptor, blocks its interaction with PD-L1 and PD-L2, and blocks PD-1 pathway-mediated immunosuppressive responses, including anti-tumor immune responses. In a syngeneic mouse tumor model, blocking PD-1 activity can inhibit tumor growth.

[0042] Tislelizumab: Tislelizumab is a humanized recombinant anti-PD-1 monoclonal antibody. The EC binding of Tislelizumab to human recombinant PD-1 50 The IC value of inhibiting the binding of PD-1 to PD-L1 is 0.12nM, the Ko value is 1.45XW10M (low density of PD-1), 1.1010nM (high density of PD-1), and the IC value of inhibiting the binding of PD-1 to PD-L1 is 50 The IC value of inhibiting the binding of PD-1 to PD-L2 is about 0.5nM. 50 About 0.4~0.6nM.

[0043] ANIKO (Peampalimab): Peampalimab is a PD-1 monoclonal antibody that uses the immunoglobulin G1 (IgG1) subtype and undergoes a modified crystal ("Fc") segment, which can durably block PD-1 / PD-L1 binding.

[0044] Sepalimab: a humanized recombinant anti-PD-1 monoclonal antibody.

[0045] The present invention provides a combination and its use in treating tumors, the use comprising providing a pharmaceutically effective amount of the composition of the present invention to a subject in a pharmaceutically acceptable manner. "Cancer" herein refers to a pathological state in the human body characterized by uncontrolled cell proliferation. Examples include, but are not limited to, cancer, lymphoma, blastoma, and leukemia. More specific examples of cancer include, but are not limited to, lung (small cell and non-small cell) cancer, breast cancer, prostate cancer, carcinoid cancer, bladder cancer, gastric cancer, pancreatic cancer, liver cancer (hepatocellular carcinoma), hepatoblastoma, colorectal cancer, head and neck cancer, squamous cell carcinoma, esophageal cancer, ovarian cancer, uterine cancer, endometrial cancer, mesothelioma, melanoma, sarcoma, osteosarcoma, liposarcoma, thyroid cancer, desmoid tumor, acute myeloid leukemia (AML) and chronic myeloid leukemia (CML).

[0046] "Therapeutically effective amount" herein refers to an amount of the compound provided herein that is effective to "treat" a disorder in a subject or mammal. In the case of treating cancer, a therapeutically effective amount of a drug can reduce the number of cancer cells, reduce the size of tumors, inhibit cancer cell infiltration into peripheral organs, inhibit tumor metastasis, inhibit tumor growth to a certain extent, and / or alleviate one or more of the symptoms associated with cancer to a certain extent.

[0047] Throughout the specification and examples, the following abbreviations will be used: PD-1: Programmed cell death protein-1 PD-L1: Programmed cell death ligand-1 UDP-UGCG: UDP glucose ceramide glucosyltransferase GCS: glucosylceramide synthase MDR1: Multidrug resistance protein 1 MHC: major histocompatibility complex IFN-γ: interferon gamma GSLs: glycosphingolipids DC: Dendritic cell GD1: Gaucher disease type 1. Example

[0048] The present invention is further illustrated by the following examples. Although preferred embodiments of the present invention are described and shown herein, it will be apparent to those skilled in the art that these embodiments are merely illustrative. Those skilled in the art may make various changes, modifications and substitutions to these embodiments without departing from the present invention. It should be understood that the optional embodiments of the present invention described herein may be used to practice the present invention. The scope of the present invention is defined by the appended claims, and methods and structures within the scope of these claims and their equivalents are also covered by the appended claims.

[0049] Example 1 Anti-tumor effect of iliglustat combined with PD-1 inhibitor (in vitro cell test model) In vitro cell models Jurkat cell line: human acute T-lymphoblastic leukemia cell line; A549 cell line: human non-small cell lung cancer cell line; Cell Counting Kit-8 (CCK-8): Dongren Chemical, Japan; Phytohemagglutinin, Aladdin; A549 cell line and Jurkat cells were cultured in 1640 medium containing 10% fetal bovine serum and 1% double antibody at 37°C and 5% CO 2 incubator.

[0050] Jurkat cell activation co-culture system: Jurkat cells were pre-treated with 2 μg / mL phytohemagglutinin for 48 h for activation.

[0051] Drugs and dosing concentrations Eliglustat, Beijing Kailai Tiancheng, dosage concentration 10 -5 M; Tislelizumab injection / Tislelizumab, BeiGene, dosing concentration 10 μg / mL; Camrelizumab for injection / Erica, Jiangsu Hengrui, dosing concentration 10 μg / mL; Experimental Grouping Normal control group (CTRL), iligliusta group (group D), tislelizumab group (group P1), carrelizumab group (group P2), iligliusta combined with tislelizumab group (P1+group D), and iligliusta combined with carrelizumab group (P2+group D).

[0052] Detection indicators Cell viability: After the cells were treated with drugs for 48 hours, the supernatant was discarded, the suspended Jurkat cells were washed with saline, and 100 μl / well of 10-fold diluted CCK8 reagent was added. After incubation at 37°C for 45 minutes, the OD value was measured at 450 nm.

[0053] Test results A549 cells were co-cultured with unactivated Jurkat cells and treated with drugs for 48 hours. Compared with the CTRL group, the viability of A549 cells in the D group / P1 group / P2 group did not change significantly, and the viability in the P2+D group was significantly reduced. There was no significant difference between the P1+D group and the P1 group, and the viability of the P2+D group was significantly reduced compared with the CTRL group, and compared with the P2 group, both were significantly reduced, with statistical differences. The experimental results are shown in the attached figure of the specification. Figure 1 : Effect of ilioglustat combined with PD-1 on A549 cell viability for 48 hours in the absence of activated Jurkat cells (n=5, compared with the CTRL group, *P<0.05). The experimental results suggest that in the absence of immune cells, ilioglustat and PD-1 alone have almost no effect on the growth of A549 non-small cell lung cancer cells, and ilioglustat combined with PD-1 antibody No. 2 has a certain anti-tumor effect.

[0054] A549 cells were co-cultured with activated Jurkat cells and treated with drugs for 48 hours. Compared with the CTRL group, the viability of A549 cells in the D group, P1+D group, and P2+D group was significantly reduced. After the combination of iliglustat and PD-1, the viability of cells in the P1+D group was significantly reduced compared with the P1 group, and the viability of cells in the P2+D group was significantly reduced compared with the P2 group. The experimental results are shown in the attached figure of the specification. Figure 2 : Effect of iliglustat combined with PD-1 on A549 cell viability for 48 hours under Jurkat cell activation (n=5, compared with CTRL group, *P<0.05). The results are consistent with the experimental results of the unactivated system, confirming that iliglustat has a significant effect on improving the activity of immune cells and can sensitize the efficacy of PD-1 inhibitors.

[0055] Example 2 Anti-tumor effect of iliglustat combined with PD-1 inhibitor (lung cancer mouse model) Material Eliglustat: Beijing Kailai Tiancheng Mouse PD-1 antibody: InVivoMAb anti-mouse PD-1, CD279, Bio X cell C57BL / 6J mice, ♀, 18-20 g, Beijing Weitonglihua Lung cancer mouse model Mouse Lewis lung adenocarcinoma tumor cells LL / 2 (LL C 1) Subcutaneously inoculated into C57BL / 6 mice, number of inoculation: 1×10 7 cells / mouse, with an inoculation volume of 0.2 ml / mouse. After 15 days, the mice were killed in a clean bench and the tumors were removed. Sterile saline was added to the homogenizer and fully ground. The tumor fluid was collected and centrifuged at 1500 rpm for 10 min. The supernatant was discarded. The cells were resuspended in sterile saline and counted. The number of cells was 1×10 7 The cells were inoculated subcutaneously on the dorsal side of the right leg of 6-week-old C57BL / 6J female mice, with an inoculation volume of 0.2 ml per mouse.

[0056] Experimental Grouping Twenty-four C57BL / 6 mice inoculated with tumor cells were randomly divided into four groups, with 6 mice in each group, namely control group (CTRL), PD-1 antibody administration group (P), iliglucositol administration group (D), and PD-1 antibody and iliglucositol combination administration group (P+D).

[0057] Dosage and administration The PD-1 antibody was prepared with sterile saline and administered immediately after use at a dose of 0.2 mg / animal; iliglucosidase was prepared with deionized water at a dose of 150 mg / kg.

[0058] The administration method was as follows: intraperitoneal injection of PD-1 antibody, 0.2 ml / mouse, once every two days, for a total of 5 times; intragastric administration of 0.1 ml / 10g body weight of iliglustat, once a day, for a total of 10 times; the control group was fed with the same volume of deionized water and intraperitoneally injected with the same volume of normal saline, and the administration began on the second day of inoculation and continued for 9 days.

[0059] Ten days after transplantation, the short diameter and long diameter of the target tumor were measured using a vernier caliper (Deli Digital Display, DL91150). The following equation was used to calculate the tumor volume TV.

[0060] Tumor volume TV (mm 3 )=long diameter (mm)×short diameter (mm)×short diameter (mm) / 2.

[0061] The tumor volumes (TV) of the control group, the iliglustat-administered group, the anti-mouse PD-1 antibody-administered group, and the combined-administered group were measured, and the values ​​obtained by logarithmic transformation of the tumor volumes were used for statistical analysis.

[0062] In the subcutaneous LL / 2 (LLc1) transplant model, the combination of iliglustat and anti-mouse PD-1 antibody showed significantly higher anti-tumor effect than either alone. The tumor growth was small in the early stage of administration, and the tumor volume increased significantly from the 8th day of administration. Compared with the CTRL group, there was no significant change in the tumor volume of mice in the P group after 6-8 days of administration, and tumor growth was inhibited on the 9th day; while the tumor volume of mice in the P+D group was significantly smaller than that of the CTRL group during the administration period, and there was a statistical difference. The daily changes in tumor volume are shown in the attached figure of the instruction manual. Figure 3 :Effects of drug administration on tumor volume of C57 lung adenocarcinoma mice (n=6, compared with CTRL group, * P <0.05, ** P <0.01, *** P <0.001).

[0063] Example 3 Anti-tumor effect of iliglustat combined with PD-1 inhibitor (liver cancer mouse model) Material Eliglustat: Beijing Kailai Tiancheng Mouse PD-1 antibody: InVivoMAb anti-mouse PD-1, CD279, Bio X cell ICR mice, ♀, 18-20 g, Beijing Weitonglihua Mouse model of liver cancer Mouse H22 liver cancer ascites cells were intraperitoneally injected into ICR mice, with the number of cells inoculated being 1×10 7 cells / mouse, inoculation volume 0.2 ml / mouse. Seven days later, mice were killed in a clean bench and ascites was extracted. The cells were centrifuged at 1500 rpm for 10 min and the supernatant was discarded. Cells were resuspended in sterile saline and counted. The number of cells was calculated as 1×10 7 The cells were inoculated subcutaneously on the dorsal side of the right leg of 6-week-old ICR mice, with an inoculation volume of 0.2 ml per mouse.

[0064] Experimental Grouping Twenty-four ICR mice inoculated with tumor cells were randomly divided into four groups, with 6 mice in each group, namely control group (CTRL), PD-1 antibody administration group (P), iliglucositol administration group (D), and PD-1 antibody and iliglucositol combination administration group (P+D).

[0065] Dosage and administration The PD-1 antibody was prepared with sterile saline and administered immediately after use at a dose of 0.2 mg / animal; iliglucosidase was prepared with deionized water at a dose of 150 mg / kg.

[0066] The administration method was as follows: intraperitoneal injection of PD-1 antibody, 0.2 ml / mouse, once every two days, for a total of 5 times; intragastric administration of 0.1 ml / 10g body weight of iliglustat, once a day, for a total of 10 times; the control group was fed with the same volume of deionized water and intraperitoneally injected with the same volume of normal saline, and the administration began on the second day of inoculation and continued for 9 days.

[0067] Ten days after transplantation, the short diameter and long diameter of the target tumor were measured using a vernier caliper (Deli Digital Display, DL91150). The following equation was used to calculate the tumor volume TV.

[0068] Tumor volume TV (mm 3 )=long diameter (mm)×short diameter (mm)×short diameter (mm) / 2.

[0069] The tumor volumes (TV) of the control group, the iliglustat-administered group, the anti-mouse PD-1 antibody-administered group, and the combined-administered group were measured, and the values ​​obtained by logarithmic transformation of the tumor volumes were used for statistical analysis.

[0070] In the H22 liver cancer ascites cell transplantation model, in vitro tumor volume measurements during the administration period showed that the tumor volume increased slightly in the early stage, but increased significantly from the 8th day of administration. The mice had no obvious tumors in the first 5 days, and the tumor volume was counted from the 6th day. Compared with the CTRL group, the tumor volume of mice in the P group / D group did not change significantly after 6-7 days of administration. Tumor growth began to be inhibited from the 8th day, and the inhibition was obvious on the 9th day; the tumor volume of mice in the P+D group was smaller after 6-9 days of administration, and there was a statistical difference. The daily changes in tumor volume are shown in the attached figure of the instruction manual. Figure 4 : Effects of drug administration on the tumor volume of ICR liver cancer mice (n=6, compared with the CTRL group, *P<0.05, **P<0.01, ***P<0.001).

Claims

1. A composition for combined cancer therapy, comprising: (i) an effective amount of a PD-1 inhibitor, and (ii) an effective amount of a GCS inhibitor.

2. The composition according to claim 1, wherein The PD-1 inhibitor is selected from: Opdivo (nivolumab), Keytruda (pembrolizumab), Toyi (toripalimab), Daboshu (sintilimab), Erica (carrelizumab), BeiGene (tislelizumab), Aniko (pembrolizumab), and Sepalizumab.

3. The composition according to claim 1, wherein The GCS inhibitor is a compound represented by Formula 1, iligrustat or a pharmaceutically acceptable salt thereof. The pharmaceutically acceptable salt of iligrustat is preferably iligrustat tartrate. Formula 1.

4. The composition according to any one of claims 1 to 3, wherein The GCS inhibitor and its pharmaceutically acceptable salt are administered once or more daily at a therapeutic dose in the form of iliglustat, and the PD-1 inhibitor is administered once every 2 weeks or once every 3 weeks at a therapeutic dose.

5. The composition according to any one of claims 1 to 4, wherein The cancer is a solid tumor or a hematological tumor.

6. The composition according to claim 5, wherein The solid tumor is non-small cell lung cancer, renal cell carcinoma, head and neck cancer, melanoma, hepatocellular carcinoma, breast cancer, endometrial cancer, ovarian cancer, gastric cancer, colorectal cancer, bladder cancer, pancreatic cancer, glioblastoma, biliary tract cancer, glioma, cervical cancer, advanced or refractory solid tumors, small cell lung cancer, squamous cell carcinoma of the lung, desmoplastic melanoma, mesothelin-positive pleural mesothelioma, esophageal cancer, anal cancer, prostate cancer and thyroid cancer, preferably non-small cell lung cancer.

7. The composition according to claim 5, wherein The blood tumor is acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, diffuse large B-cell lymphoma, EBV-positive DLBCL, primary mediastinal large B-cell lymphoma, T cell / histiocyte-rich large B-cell lymphoma, follicular lymphoma, Hodgkin's disease lymphoma, mantle cell lymphoma, multiple myeloma, myeloid leukemia-1 protein, myelodysplastic syndrome, non-Hodgkin's lymphoma or small lymphocytic lymphoma, preferably acute lymphocytic leukemia and multiple myeloma.

8. A method of combined cancer treatment, comprising simultaneously administering a GCS inhibitor and a PD-1 inhibitor to an individual.

9. The method of claim 8, further comprising one or more additional therapeutic agents, preferably chemotherapeutic agents, biotherapeutic agents, immunogenic agents other than the GCS inhibitor.

10. The method according to claim 8 or 9, wherein: The method can be used before or after surgery, and before, during or after radiation therapy to remove a tumor.

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