Application of TGF-beta RI inhibitor in tumor treatment and pharmaceutical composition of TGF-beta RI inhibitor

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to application of a TGF-beta RI inhibitor in tumor treatment and a pharmaceutical composition of the TGF-beta RI inhibitor. The invention discloses an application of a TGF-beta RI inhibitor single drug or a drug combination in tumor treatment, and the TGF-beta RI inhibitor single drug or the drug combination has a good anti-tumor activity effect, can obviously improve the health condition and the life quality of a patient, and is beneficial to be used as a treatment scheme of advanced or metastatic tumors.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the use of a TGF-βRI inhibitor in treating tumors. Background Art

[0002] Transforming growth factor-β (TGF-β) is a multifunctional growth factor superfamily with a wide range of biological activities, participating in early embryonic development, cartilage and bone formation, extracellular matrix synthesis, inflammation, interstitial fibrosis, regulation of immune and endocrine functions, and tumor formation and development. TGF-β signaling molecules transduce signals through transmembrane receptor complexes. TGF-β receptors are transmembrane proteins present on the cell surface, which are divided into type I receptors (TGF-βRI), type II receptors (TGF-βR II), and type III receptors (TGF-βRIII).

[0003] Current studies have shown that in tumor cells, TGF-β can directly affect tumor growth (non-intrinsic effects of TGF-β signals), or indirectly affect tumor growth (intrinsic effects of TGF-β) by inducing epithelial-mesenchymal transition, blocking anti-tumor immune responses, increasing tumor-associated fibrosis, and enhancing angiogenesis. At the same time, TGF-β has a strong fibrosis-inducing effect, and it is an activator of tumor-associated fibroblasts. These fibroblasts are the main source of collagen type I and other fibrotic factors. The induced products of fibroblasts and other fibrotic factors may continue to cultivate a microenvironment that reduces immune responses, increases drug resistance, and enhances tumor angiogenesis. In addition, during individual development and tumor growth, TGF-β affects angiogenesis. For example, mouse embryos with TGF-βRI deficiency show severe vascular development defects, proving that the TGF-β signaling pathway is a key regulator in the development of vascular endothelial and smooth muscle cells. In 2013, the FDA granted Eli Lilly's small molecule TGF-βR I inhibitor LY2157299 (WO 2002 / 094833) for the treatment of glioma and liver cancer. WO2017215506A1 discloses a series of TGF-βRI inhibitors, which show good tumor suppression activity, but still need to develop effective doses and dosing regimens, and further optimize single-drug or combined treatment regimens, in order to further improve clinical benefits and improve quality of life. Summary of the invention

[0004] One aspect of the present invention provides a use of a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of an anti-tumor drug.

[0005]

[0006] Another aspect of the present invention provides a method for treating tumors, comprising the step of administering a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0007]

[0008] In some embodiments, the tumor is selected from pelvic paraganglioma, small cell lung cancer, non-small cell lung cancer, nasopharyngeal carcinoma, gastric cancer, intestinal cancer, colon cancer, cervical cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumors, ovarian tumors, peritoneal tumors, melanoma, glioma, glioblastoma, hepatocellular carcinoma, papillary nephroma, head and neck squamous cell carcinoma, esophageal squamous cell carcinoma, endometrial cancer, synovial sarcoma, thymic carcinoma, bile duct cancer, thyroid cancer, bile duct cancer / gallbladder cancer, colorectal cancer, urothelial carcinoma, esophageal cancer, gastric and esophageal junction adenocarcinoma, pleural mesothelioma or Hodgkin lymphoma.

[0009] Another aspect of the present invention provides a pharmaceutical combination or kit comprising a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapeutic agent.

[0010]

[0011] In some embodiments, the pharmaceutical combination or kit is used to treat a tumor.

[0012] Another aspect of the present invention provides use of the aforementioned drug combination or drug kit in the preparation of anti-tumor drugs.

[0013] Another aspect of the present invention provides a method for treating tumors, comprising the step of administering a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapeutic agent to a subject in need thereof.

[0014] In some embodiments, the tumor is an advanced or metastatic tumor that has failed at least standard treatment, or cannot be treated with standard treatment due to intolerable toxicity, or has no standard treatment.

[0015] In some embodiments, the tumor is a solid tumor.

[0016] In another aspect of the present invention, a medicine kit is provided, which comprises a container, a pharmaceutical composition contained in the container, and a package insert, wherein the package insert contains instructions for administering the compound of formula I or a pharmaceutically acceptable salt thereof, an immunosuppressant, or a combination of the two.

[0017] The TGF-βRI inhibitor of the present invention is used alone or in combination with an immunotherapeutic agent, or is used in combination with an immunotherapeutic agent and concurrently with chemotherapy (CT), radiotherapy (RT) or chemoradiotherapy (CRT) for the treatment of tumors. It has good anti-tumor activity. In some tumor types, the objective response rate (ORR) is as high as 66.7%, and the disease control rate (DCR) reaches 100%, which significantly improves the health status and quality of life of the subjects and provides a new treatment option for advanced or metastatic tumors.

[0018] The compound of formula I of the present invention or its pharmaceutically acceptable salt is a specific small molecule inhibitor of transforming growth factor-β receptor I (TGF-βRI). Through the inhibitory effect on the TGF-β signaling pathway, it has multiple anti-tumor effects of anti-tumor angiogenesis, inducing tumor cell apoptosis and enhancing anti-tumor immunity, and has a significant growth inhibitory effect on a variety of tumors. In the mouse H22 liver cancer model, the tumor inhibition rate (TGI) of the compound of formula I of the present invention or its pharmaceutically acceptable salt at a dose of 15 mg / kg orally twice a day is 61.6%; and the compound of formula I of the present invention or its pharmaceutically acceptable salt can be used in combination with programmed death receptor 1 (PD-1) immune checkpoint inhibitors in the Pan02 pancreatic cancer mouse model to significantly inhibit tumor growth, show a synergistic effect, and take effect at a lower dose.

[0019] Immune escape is a key strategy for tumor cells to escape immune system surveillance while promoting their survival, metastasis, and spread. In fact, tumors have developed a variety of mechanisms to suppress immune responses, including immune checkpoint molecules such as PD-1. PD-1 is a cell surface inhibitory receptor that plays an important physiological role in maintaining peripheral tolerance. Under pathological conditions, tumor cells upregulate the expression of PD-1, activate the PD-L1-PD-1 signaling pathway, and block the activation of immune cells, which is a mechanism for tumor cells to escape anti-tumor immune control. Targeting PD-1 is a major breakthrough in tumor immunotherapy. In fact, the success of PD-1 blocking immunotherapy represents an unprecedented success in treating different cancer types. In order to improve the treatment effect, a deeper understanding of the mechanisms regulating PD-1 expression and signaling in tumors is needed. In recent years, a large number of studies have shown that in patients with a variety of solid tumors (such as pelvic paraganglioma, small cell lung cancer, non-small cell lung cancer, nasopharyngeal carcinoma, gastric cancer, intestinal cancer, colon cancer, cervical cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumors, ovarian tumors, peritoneal tumors, melanoma, glioma, glioblastoma, hepatocellular carcinoma, papillary nephroma, head and neck squamous cell carcinoma, esophageal squamous cell carcinoma, endometrial cancer, synovial sarcoma, thymic carcinoma, bile duct cancer, thyroid cancer, bile duct cancer / gallbladder cancer, colorectal cancer, urothelial carcinoma, esophageal cancer, gastric and esophageal junction adenocarcinoma, pleural mesothelioma or Hodgkin's lymphoma, etc.), PD-1 was found to be highly expressed in tumor-infiltrating CD8+T cells in different proportions and was negatively correlated with the disease. Immunotherapy has achieved unprecedented breakthroughs in the treatment of different tumors by blocking and destroying PD-1 / PD-L1 signals, which has led to a real revolution in the treatment of highly aggressive cancers. Currently, multiple PD-1 antagonists have been approved for the treatment of solid tumors. For example, nivolumab has been approved for multiple indications, including adult patients with locally advanced or metastatic non-small cell lung cancer (NSCLC), patients with recurrent or metastatic squamous cell carcinoma of the head and neck (SCCHN), patients with advanced or metastatic gastric cancer, gastroesophageal junction cancer, or esophageal adenocarcinoma, adult patients with non-epithelioid malignant pleural mesothelioma, and patients with esophageal cancer or gastroesophageal junction cancer. For example, pembrolizumab has been approved for use in patients with non-small cell lung cancer, esophageal cancer, recurrent head and neck squamous cell carcinoma (HNSCC), metastatic or unresectable MSI-H / dMMR colorectal cancer, advanced unresectable or metastatic esophageal or gastroesophageal junction cancer, and hepatocellular carcinoma (HCC).For example, sintilimab is currently approved for the treatment of relapsed or refractory classical Hodgkin lymphoma, advanced or metastatic squamous NSCLC, unresectable or metastatic hepatocellular carcinoma, advanced, recurrent or metastatic esophageal squamous cell carcinoma, gastric or gastroesophageal junction adenocarcinoma, locally advanced or metastatic non-squamous non-small cell lung cancer (NSCLC). Tislelizumab is currently approved for the treatment of relapsed / refractory classical Hodgkin lymphoma (r / r cHL), locally advanced or metastatic urothelial carcinoma, advanced lung squamous cell carcinoma, advanced nasopharyngeal carcinoma, advanced or metastatic non-small cell lung cancer (NSCLC), adult patients with advanced unresectable or metastatic MSI-H / dMMR solid tumors, locally advanced or metastatic esophageal squamous cell carcinoma (ESCC), recurrent or metastatic nasopharyngeal carcinoma (NPC), and advanced unresectable or metastatic gastric or gastroesophageal junction adenocarcinoma (G / GEJ).

[0020] Based on the above research on the molecular mechanism of the TGF-β1 signaling pathway and the immunotherapy mechanism of PD-1 antagonists, the compound of formula I of the present invention or its pharmaceutically acceptable salt or its combination with a PD-1 antagonist is expected to have great clinical value in the treatment of pelvic paraganglioma, small cell lung cancer, bile duct cancer, thymic carcinoma, nasopharyngeal carcinoma, colon cancer, thyroid cancer, cervical cancer, head and neck squamous cell carcinoma, colorectal cancer, hepatocellular carcinoma, esophageal squamous cell carcinoma, endometrial cancer, synovial sarcoma, non-small cell lung cancer, gastric cancer, esophageal cancer, gastric and esophageal junction adenocarcinoma, melanoma, urothelial carcinoma, pleural mesothelioma, Hodgkin's lymphoma and other tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The graph shows the changes in tumor growth in mice bearing Pan02 homologous transplant tumor model in Example 1 after administration of different test substances (data points represent the mean and standard error within the group, n=7).

[0022] Figure 2 The graph shows the changes in relative tumor growth in mice bearing Pan02 homologous transplant tumor model in Example 1 after administration of different test substances (data points represent the mean and standard error within the group, n=7).

[0023] Figure 3 The tumor growth curves of each group in Example 2 are shown (mean ± standard error).

[0024] Figure 4 The tumor weight of each group in Example 2 is shown (mean ± standard error).

[0025] Note: The compound of formula I marked in each figure is the p-toluenesulfonate salt of the compound of formula I. DETAILED DESCRIPTION

[0026] Compounds of formula I

[0027] The pharmaceutically acceptable salt of the compound of formula I described in the present invention is selected from benzenesulfonate, p-toluenesulfonate, methanesulfonate, ethanesulfonate, hydrochloride or sulfate.

[0028] In some embodiments, the pharmaceutically acceptable salt of the compound of Formula I is p-toluenesulfonate.

[0029] The administration frequency of the compound of formula I or a pharmaceutically acceptable salt thereof is once / day, twice / day, three times / day, once / two days, once / three days, once / week, once / two weeks, once / three weeks, continuous administration for one week and rest for one week, or continuous administration for two weeks and rest for two weeks. When the administration is continuous for one week or continuous for two weeks, the daily administration frequency may be once / day, twice / day or three times / day.

[0030] The compound of formula I or a pharmaceutically acceptable salt thereof can be administered in any amount or manner that elicits the desired response in a subject, for example, 15-100 mg / kg can be administered twice daily to mice or at equivalent dosing amounts to other subjects.

[0031] The dosage of the compound of formula I or a pharmaceutically acceptable salt thereof is selected from 5-100 mg / kg, preferably 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 65 mg / kg, 70 mg / kg, 75 mg / kg, 80 mg / kg, 85 mg / kg, 90 mg / kg, 95 mg / kg or 100 mg / kg.

[0032] The daily dosage of the compound of formula I or a pharmaceutically acceptable salt thereof is about 5-500 mg, for example, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg or about 500 mg.

[0033] In some embodiments, the compound of Formula I or a pharmaceutically acceptable salt thereof is administered continuously to the subject.

[0034] In some embodiments, the compound of Formula I or a pharmaceutically acceptable salt thereof is administered intermittently to the subject.

[0035] In some embodiments, the continuous administration of a compound of Formula I, or a pharmaceutically acceptable salt thereof, is interrupted by one or more drug holidays.

[0036] In some embodiments, each drug holiday lasts about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days.

[0037] In some embodiments, the compound of Formula I, or a pharmaceutically acceptable salt thereof, is administered daily for one week, followed by a one-week drug holiday.

[0038] In some embodiments, the compound of Formula I, or a pharmaceutically acceptable salt thereof, is administered daily for two weeks, followed by a two-week drug holiday.

[0039] In some embodiments, when the compound of Formula I or a pharmaceutically acceptable salt thereof is used to treat a tumor, the compound is continuously cycled between a two-week dosing period and a one-week drug-free period until a change in the disease state is observed.

[0040] In some embodiments, the change in disease state is a complete remission.

[0041] In some embodiments, the change in disease state is a partial remission.

[0042] The compound of formula I or its pharmaceutically acceptable salt is in the form of a pharmaceutical composition suitable for oral administration.

[0043] In some embodiments, the pharmaceutical composition comprises a compound of formula I or a pharmaceutically acceptable salt thereof as an active ingredient and at least one pharmaceutically acceptable excipient, wherein the pharmaceutically acceptable excipient is selected from one or more of a filler, a disintegrant, a lubricant, and a glidant.

[0044] The filler is selected from one or more of cellulose, silicified microcrystalline cellulose, starch, calcium hydrogen phosphate, sucrose, lactose, mannitol, xylitol, lactitol, and maltodextrin.

[0045] The disintegrant is selected from one or more of sodium starch glycolate, sodium carboxymethyl starch, corn starch, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, cross-linked carboxymethyl cellulose, methyl cellulose, pregelatinized starch, and sodium alginate.

[0046] The lubricant is selected from one or more of stearic acid, magnesium stearate, sodium stearate, calcium stearate, zinc stearate, glyceryl monostearate, glyceryl distearate, glyceryl tristearate, myristic acid, palmitic acid, sodium stearyl fumarate, talc, hydrogenated vegetable oil, and mineral oil.

[0047] The glidant is selected from one or more of powdered cellulose, magnesium trisilicate, colloidal silicon dioxide, silicon dioxide, and talc.

[0048] In some embodiments, the weight percentage of the active ingredient of the compound of formula I or a pharmaceutically acceptable salt thereof is 10%-50% active ingredient, preferably 15%-30%.

[0049] The weight percentage of the filler is 10%-80%, preferably 60%-80%.

[0050] The weight percentage of the disintegrant is 1%-10%, preferably 2%-8%.

[0051] The weight percentage of the lubricant is 1%-5%, preferably 1%-3%.

[0052] The weight percentage of the glidant is 0.01%-5%, preferably 0.5-2%.

[0053] In some embodiments, the pharmaceutical composition is obtained by dissolving the compound of formula I or a pharmaceutically acceptable salt thereof in a solvent.

[0054] In some embodiments, the solvent is an aqueous solution of methylcellulose and / or an aqueous solution of Tween-80; preferably an aqueous solution of 0.5% methylcellulose and / or an aqueous solution of 0.5% Tween-80.

[0055] Immunosuppressants

[0056] The immunosuppressant is a therapeutic agent (any one) or a combination thereof against the following targets: PD-1, PD-L1, CTLA-4, PD-L2, LAG3, TIM3, 2B4, A2aR, B7H1, B7H3, B7H4, BTLA, CD2, CD27, CD28, CD30, CD40, CD70, CD80, CD86, CD137, CD160, CD226, CD276, DR3, GAL9, GITR, HAVCR2, HVEM, IDO1, IDO2, ICOS, KIR, LAIR1, LIGHT, MARCO (macrophage receptor with collagen structure), PS (phosphatidylserine), OX-40, SLAM, TIGHT, VISTA or VTCN1.

[0057] In some embodiments, the immunotherapeutic agent is selected from Galunisertib (LY2157299), Toripalimab, AMP-224, Zimberelimab (GLS-010), Sintilimab (IBI-308), Tislelizumab (BGB-A317), Pidilizumab, Pembrolizumab, LZM-009, Nivolumab, Sintilimab, Cemiplimab, Tislelizumab, Dostarlimab, BI-754091, or Spartalizumab (PDR-001).

[0058] The administration frequency of the immunotherapy agent is once / two days, once / three days, twice / week, once / week, once / two weeks, once / three weeks, once / month, once / two months, administer for one week and rest for one week, or administer for two weeks and rest for two weeks. When administering for one week and resting for one week, the administration frequency can be once / week, and when administering for two weeks and resting for two weeks, the administration frequency can be once / week or once / two weeks.

[0059] The dosage of the immunotherapeutic agent is selected from 1-20 mg / kg, for example, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg.

[0060] In some embodiments, the immunotherapeutic agent is an anti-PD-1 antibody or an antigen-binding fragment thereof.

[0061] In some embodiments, the immunotherapeutic agent is in a formulation suitable for administration by injection.

[0062] The immunotherapeutic agent may be administered in any amount or manner that elicits the desired response in a subject, for example, 10 mg / kg may be administered twice weekly to mice or an equivalently effective dosing amount to other subjects.

[0063] In some embodiments, the immunotherapeutic agent is administered continuously to the subject.

[0064] In some embodiments, the immunotherapeutic agent is administered intermittently to the subject.

[0065] In some embodiments, continuous administration of an immunotherapeutic agent is interrupted by one or more drug holidays.

[0066] In some embodiments, each drug holiday lasts about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days.

[0067] Medication combinations or kits

[0068] The present invention provides a drug combination or a drug kit comprising a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapeutic agent, and a method for treating tumors by combining a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapeutic agent.

[0069] The combined administration route of the present invention can be selected from oral administration and parenteral administration, respectively. The parenteral administration includes, but is not limited to, intraperitoneal injection, intravenous injection, subcutaneous injection, and intramuscular injection.

[0070] When the compound of formula I or its pharmaceutically acceptable salt and the immunotherapeutic agent are used to treat tumors, there is no restriction on the order of administration of the two. They can be administered simultaneously, the immunotherapeutic agent can be administered first and then the compound of formula I, or the immunotherapeutic agent can be administered first and then the anti-PD-1 antibody.

[0071] The pharmaceutical combination or kit comprises one cycle or two cycles of dosage of the compound of formula I or a pharmaceutically acceptable salt thereof or an immunotherapeutic agent.

[0072] In one embodiment, the pharmaceutical combination or kit comprises 1-100 mg; e.g., 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg of a compound of Formula I.

[0073] In one embodiment, the pharmaceutical combination or kit comprises 1-300 mg, e.g., 3 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg of an immunotherapeutic agent.

[0074] The immunotherapeutic agent is preferably a PD-1 antibody, more preferably BP0146 (BioXcell) or Teplizumab.

[0075] The medicine kit comprises a container, a drug combination contained in the container, and a package insert, wherein the package insert contains instructions for administering the compound of formula I or its pharmaceutically acceptable salt and the immunosuppressant according to the aforementioned method.

[0076] Combined with other cancer treatment options

[0077] In one embodiment of the present invention, the combination therapy of the compound of formula I or a pharmaceutically acceptable salt thereof and an immunotherapeutic agent can also be combined with other cancer treatment regimens.

[0078] In some embodiments, the other cancer treatment regimen includes chemotherapy (CT), radiotherapy (RT) or chemoradiotherapy (CRT) for the treatment of tumors. Wherein the chemotherapy, radiotherapy or chemoradiotherapy are given to the subject in need of treatment simultaneously, successively or at intervals.

[0079] In some embodiments, the chemotherapy drugs include, but are not limited to, one or more of alkylating agents, podophyllums, camptothecins, taxanes, antimetabolites, and antibiotic anti-tumor drugs.

[0080] Preferably, the chemotherapy drug can be one or more of etoposide, doxorubicin, topotecan, irinotecan, fluorouracil, gemcitabine, paclitaxel, platinum drugs, and anthracyclines, for example, paclitaxel, carboplatin, cisplatin, or pemetrexed.

[0081] In some embodiments, the chemotherapeutic drug is administered in a once-daily dosing, continuous dosing regimen.

[0082] In some embodiments, radiation therapy can be treatment with electrons, photons, protons, alpha (α) emitters, other ions, radionucleotides, boron captured neutrons, and combinations thereof.

[0083] In some embodiments, the radiation therapy is administered according to the location of the lesion and actual clinical conditions, using conventional clinical radiation therapy doses.

[0084] In some embodiments, the radiation therapy uses the following doses (α / β=10): brain BED: 60-80 Gy; or lung lesions BED: 80-180 Gy; or bone metastasis BED: 60-80 Gy; or liver metastasis BED: 60-120 Gy; or retroperitoneal lymph nodes, or mediastinal lymph nodes: 60-120 Gy; or metastasis to other parts: 60-120 Gy.

[0085] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof and an immunotherapeutic agent are combined and further used for tumor treatment by chemotherapy (CT), radiotherapy (RT) or chemoradiotherapy (CRT), and the three can be administered in the same administration cycle. The administration cycle can be 1 week, 2 weeks, 3 weeks (21 days), 3-4 weeks (21-28 days), 4 weeks (28 days), 6 weeks, 3 months, 6 months, 9 months, 1 year, 18 months or 24 months.

[0086] Anti-tumor

[0087] The tumor described in the present invention may be a solid tumor or a hematological tumor.

[0088] In some embodiments, the tumor is selected from solid tumors. For example, the solid tumor is selected from pelvic paraganglioma, small cell lung cancer, non-small cell lung cancer, nasopharyngeal carcinoma, gastric cancer, intestinal cancer, colon cancer, breast cancer, cervical cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumors, ovarian tumors, peritoneal tumors, melanoma, glioma, glioblastoma, hepatocellular carcinoma, papillary nephroma, head and neck squamous cell carcinoma, esophageal squamous cell carcinoma, endometrial cancer, synovial sarcoma, thymic carcinoma, bile duct cancer, thyroid cancer, bile duct cancer / gallbladder cancer, colorectal cancer, urothelial carcinoma or esophageal cancer.

[0089] The compound of formula I or a pharmaceutically acceptable salt thereof is used in combination with an immunosuppressant for anti-tumor treatment, wherein the tumor is selected from pelvic paraganglioma, small cell lung cancer, non-small cell lung cancer, nasopharyngeal carcinoma, gastric cancer, intestinal cancer, colon cancer, breast cancer, cervical cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumors, ovarian tumors, peritoneal tumors, melanoma, glioma, glioblastoma, hepatocellular carcinoma, papillary nephroma, head and neck squamous cell carcinoma, esophageal squamous cell carcinoma, endometrial cancer, synovial sarcoma, thymic carcinoma, bile duct cancer, thyroid cancer, bile duct cancer / gallbladder cancer, colorectal cancer, urothelial carcinoma, esophageal cancer, gastric and esophageal junction adenocarcinoma, pleural mesothelioma or Hodgkin's lymphoma.

[0090] I. Definition and Explanation

[0091] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered to be uncertain or unclear in the absence of a special definition, but should be understood according to its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or its active ingredient.

[0092] As used herein and unless otherwise specified, the terms "comprising", "including", "having", "containing", including grammatical equivalents thereof, should generally be understood as open and non-limiting, for example, not excluding other unlisted elements or steps.

[0093] The term "inhibit" is used relative to a control. One skilled in the art will readily determine the appropriate control for each experiment. For example, a reduced response in a subject or cell treated with a compound is compared to a response in a subject or cell not treated with the compound.

[0094] The disclosure of all ranges in the present invention shall be deemed as the disclosure of all sub-ranges and all point values ​​within the ranges. For example, the disclosure of 5-100 shall be deemed as also disclosing the ranges of 5-50, 50-100, etc., and also disclosing point values ​​of 5mg, 10mg, 20mg, 30mg, 40mg, 50mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, etc.

[0095] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0096] The compounds may be present in the pharmaceutical compositions as pharmaceutically acceptable salts.

[0097] The term "pharmaceutically acceptable salt" refers to salts of compounds of the invention, prepared from compounds of the invention having specific substituents with relatively nontoxic acids or bases. When the compounds of the invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When the compounds of the invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, fumaric acid and methanesulfonic acid, and also include salts of amino acids (such as arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups, and thus can be converted into any base or acid addition salt.

[0098] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, these compounds in free acid or base form are reacted with a stoichiometric amount of an appropriate base or acid to prepare.

[0099] The term "effective amount" or "therapeutically effective amount" encompasses an amount sufficient to ameliorate or prevent the symptoms or conditions of a medical condition. An effective amount also means an amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on factors such as the condition to be treated, the patient's overall health, the method, route and dosage of administration, and the severity of side effects. An effective amount may be the maximum dose or dosing regimen that avoids significant side effects or toxic effects.

[0100] The amount of compound administered may depend on the subject being treated, the subject's age, health, sex, and weight, the type of concurrent treatment (if any), the severity of the condition, the nature of the desired effect, the manner and frequency of treatment, and the judgment of the prescribing physician. The frequency of administration may also depend on the pharmacodynamic effect on arterial oxygen tension. However, the most preferred dosage may be adjusted for individual subjects, as will be understood by those skilled in the art and can be determined without undue experimentation. This generally includes adjusting the standard dosage (e.g., if the patient is of low weight, then reducing the dosage).

[0101] The "combined" administration method of the present invention is selected from simultaneous administration, independent formulation and co-administration or independent formulation and sequential administration.

[0102] The term "antibody" herein is used in the broadest sense and specifically encompasses monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, as long as they exhibit the desired biological activity. Antibodies can be murine, human, humanized, chimeric, or derived from other species. Antibodies are proteins produced by the immune system that are capable of recognizing and binding to specific antigens. Target antigens typically have multiple binding sites, also known as epitopes, which are recognized by CDRs (complementarity determining regions) on a variety of antibodies. Each antibody that specifically binds to a different epitope has a different structure. Therefore, an antigen may have more than one corresponding antibody. Antibodies include full-length immunoglobulin molecules or immunologically active portions of full-length immunoglobulin molecules, i.e., molecules that contain antigen binding sites that immunospecifically bind to a target antigen or a portion thereof, and these targets include, but are not limited to, cancer cells or cells that produce autoimmune antibodies associated with autoimmune diseases.

[0103] The term "humanized antibody" refers to a non-human antibody that has been genetically engineered, and its amino acid sequence has been modified to increase the homology with the sequence of a human antibody. Generally speaking, all or part of the CDR region of a humanized antibody comes from a non-human antibody (donor antibody), and all or part of the non-CDR region (e.g., variable region FR and / or constant region) comes from a human immunoglobulin (recipient antibody). Humanized antibodies generally retain or partially retain the expected properties of the donor antibody, including but not limited to, antigen specificity, affinity, reactivity, ability to increase immune cell activity, ability to enhance immune response, etc.

[0104] The term "subject" refers to an animal, including but not limited to primates (e.g., humans), monkeys, cows, pigs, sheep, goats, horses, dogs, cats, rabbits, rats or mice. The terms "subject" and "patient" are used interchangeably herein with respect to, for example, mammalian subjects, such as humans. The term "antigen-binding fragment" herein refers to a Fab fragment, a Fab' fragment, a F(ab')2 fragment, and an Fv fragment sFv fragment that binds to human PD-1, which has antigen-binding activity.

[0105] The term "CR" in this article means complete remission.

[0106] The term “PR” in this article means partial response.

[0107] The term "SD" herein means stable disease.

[0108] The term "PD" in this article refers to progressive disease.

[0109] The term "ORR" in this article, objective response rate, complete response (CR) + partial response (PR), refers to the objective tumor efficacy assessed by the RECIST version 1.1 standard, and refers to the percentage of CR and PR cases among patients who can be evaluated for efficacy.

[0110] The term "DCR" in this article, disease control rate, complete remission (CR) + partial remission (PR) + stable disease (SD), refers to the percentage of cases with CR, PR and SD (≥4 weeks) in patients who can be evaluated for efficacy. For example, disease control rate (DCR): defined as the proportion of subjects with complete remission (CR), partial remission (PR) and stable disease (SD) in the total number of subjects, that is, DCR = (number of subjects with CR + PR + SD) / total number of subjects * 100%, binomial distribution is used to estimate the DCR of the treatment group and the control group and its 95% CI, as well as the difference between the groups and its 95% CI; for example, duration of remission (DOR): defined as the time interval from the first recorded disease remission to disease progression or death (whichever occurs first), Kaplan-Meier is used to estimate the median DOR, and a survival curve is drawn.

[0111] As used herein, the term "radiation therapy" or "radiotherapy" refers to the use of ionizing radiation to treat cancer.

[0112] The term "chemotherapy" as used herein refers to the use of specific chemical agents to treat cancer.

[0113] II. Embodiment

[0114] Example 1: The in vivo efficacy of the p-toluenesulfonate salt of the compound of formula I and its combination with PD-1 antibody in a subcutaneous homograft tumor model of the mouse pancreatic cancer cell line Pan02 was evaluated.

[0115] Experimental Materials:

[0116] C57BL / 6 mice, 8-10 weeks old, female, were purchased from Shanghai Slake Laboratory Animal Co., Ltd. They were kept in an SPF animal room with a temperature of 20-26°C and a humidity of 40-70%.

[0117] PD-1 antibody: Catalog number: BP0146, purchased from BioXcell.

[0118] Mouse pancreatic cancer Pan02 cells were purchased from the National Cancer Institute (NCI) of the United States and cultured in vitro in a monolayer culture medium containing 10% fetal bovine serum in a 5% CO2 incubator at 37°C. The cells were routinely digested and passaged using trypsin-EDTA. When the cells were in the exponential growth phase and the saturation was 80%-90%, the cells were harvested and counted.

[0119] Experimental steps:

[0120] Cell seeding and grouping:

[0121] The cells were resuspended in DPBS (Dulbecco's Phosphate-Buffered Saline) at a density of 5×10 7 cells / ml. 0.1mL DPBS (containing 5×10 6 Pan02 cells) were subcutaneously inoculated on the right back of each mouse until the average tumor volume reached about 61 mm 3 The patients were randomly divided into groups and given medication according to the tumor volume. The medication schedule is shown in Table 1.

[0122] Table 1 Dosage regimen

[0123] Group Test article Number of animals / group Route of administration Dosage mg / kg Dosing frequency 1 Vehicle control group 7 po / BID 2 PD-1 Antibodies 7 ip 10 BIW 3 p-Toluenesulfonate of the compound of formula I 7 po 50 BID 4 p-Toluenesulfonate of the compound of formula I + PD-1 7 po+ip 25+10 BID+BIW 5 p-Toluenesulfonate of the compound of formula I + PD-1 7 po+ip 50+10 BID+BIW

[0124] BID: twice daily; BIW: twice weekly; po: oral; ip: intraperitoneal.

[0125] Preparation of solvent and test substance

[0126] A. Solvents for each group / compound

[0127] a) Vehicle control group: 0.5% MC (Methylcellulose) + 0.5% Tween-80

[0128] b) p-Toluenesulfonate of the compound of formula I: 0.5% MC + 0.5% Tween-80

[0129] c) PD-1 antibody: DPBS

[0130] B. Preparation method

[0131]

[0132] Daily animal observation

[0133] The health status and mortality of the animals were monitored daily. Routine inspections included observing the effects of tumor growth and drug treatment on the daily behavior of the animals, such as behavioral activities, food and water intake (only visual observation), weight changes, physical signs or other abnormal conditions. The number of deaths and side effects of animals in each group were recorded based on the number of animals in each group.

[0134] Experiment termination

[0135] If the animal's health condition continues to deteriorate, or the tumor volume exceeds 2,000mm 3 If the animal is seriously ill or in pain, it must be euthanized. If the animal has any of the following conditions, notify the veterinarian and euthanize it:

[0136] Obvious weight loss, with a weight loss of more than 20%;

[0137] No free access to food and water;

[0138] The average tumor volume of the control group reached 2,000mm 3 , the experiment terminated.

[0139] The animals developed the following clinical manifestations and continued to deteriorate:

[0140] Piloerection; arched back; pale ears, nose, eyes or feet; rapid breathing; convulsions; continuous diarrhea; dehydration; slow movement; vocalization.

[0141] Tumor measurements and experimental parameters

[0142] Tumor diameter was measured with a vernier caliper twice a week. The tumor volume was calculated as follows: V = 0.5 × a × b 2 , a and b represent the long and short diameters of the tumor, respectively.

[0143] The anti-tumor efficacy of the compound was evaluated by tumor growth inhibition rate TGI (%) or relative tumor proliferation rate T / C (%).

[0144] T / C(%)=T RTV / C RTV ×100(T RTV : Average RTV of the treatment group; C RTV : Average RTV of negative control group).

[0145] The relative tumor volume (RTV) was calculated based on the results of tumor measurement. The calculation formula is:

[0146] RTV=V t / V0, where V0 is the tumor volume measured at the time of group administration (i.e., D0), V t is the tumor volume at a certain measurement, T RTV With C RTV Get data from the same day.

[0147] TGI (%) reflects the tumor growth inhibition rate.

[0148] TGI (%) = [(1-(average tumor volume of a treatment group at the end of drug administration-average tumor volume of the treatment group at the beginning of drug administration)) / (average tumor volume of the solvent control group at the end of treatment-average tumor volume of the solvent control group at the beginning of treatment)]×100.

[0149] Collection method

[0150] At the end of the experiment, tumors from all mice were collected, snap-frozen in liquid nitrogen, and stored at -80°C.

[0151] Statistical analysis

[0152] Statistical analysis was performed using SPSS software based on the relative tumor volume and tumor weight at the end of the experiment. The comparison between two groups was analyzed using t-test, and the comparison between three or more groups was analyzed using one-way ANOVA (Tukey HSD). If the variance was not equal (F value was significantly different), the Games-Howell method was used for testing. P<0.05 was considered to be significantly different.

[0153] Experimental results:

[0154] Effects of the test substance on tumor growth

[0155] In this experiment, the p-toluenesulfonate salt of the compound of formula I and its combination with PD-1 were evaluated in vivo in a mouse pancreatic cancer cell Pan02 homograft tumor model. Figure 1 and Figure 2 shown.

[0156] The experiment ended on the 31st day, and the tumors of each mouse were collected, weighed, and photographed. The tumor weight and its analysis are shown in Tables 2 and 3. The tumor weight and tumor volume data have the same trend. The results of the tumor weight data analysis are consistent with the results of the tumor volume analysis. The results showed that compared with the solvent control group, the p-toluenesulfonate salt of the compound of Formula I + PD-1 group can significantly inhibit the growth of pancreatic cancer cell Pan02 homograft tumors in mice.

[0157] Table 2 Antitumor effect of the test substance on Pan02 homograft tumor model (based on tumor volume data on day 31 after administration)

[0158]

[0159]

[0160] Note:

[0161] a. Mean ± SEM, n = 7.

[0162] b. Relative tumor proliferation rate T / C (%) = T RTV / C RTV ×100(T RTV : Average RTV of the treatment group; C RTV : Average RTV of negative control group).

[0163] c. Tumor (volume) growth inhibition rate TGI (%) = [(1-(average tumor volume of a treatment group at the end of drug administration - average tumor volume of the treatment group at the beginning of drug administration)) / (average tumor volume of the solvent control group at the end of treatment - average tumor volume of the solvent control group at the beginning of treatment)] × 100.

[0164] Table 3 Effects of the test substances on tumor weight in mice with Pan02 homologous transplanted tumors

[0165]

[0166] Note:

[0167] a. Mean ± SEM, n = 7.

[0168] b. Tumor (body weight) growth inhibition by T / C weight =(TW 溶媒 -TW treatment ) / TW 溶媒 Calculation (TW 溶媒 : Tumor weight of vehicle control group, TW treatment : tumor weight of drug-treated group).

[0169] Effects of the test substance on the body weight of mice bearing subcutaneous transplanted pancreatic cancer Pan02 cells

[0170] The experiment was carried out according to the dosing regimen, and the animals' activities such as eating and drinking were observed every day, and the weight of the animals was recorded every two or three days. During the entire dosing cycle, the mice's food intake and drinking were normal. During the experiment, the weight of mice in each group increased steadily, and no abnormality was observed in the mice. This indicates that the test substance has no obvious toxicity to C57BL / 6 mice at the experimental dose.

[0171] Experimental conclusion:

[0172] In this experiment, the mouse pancreatic cancer cell Pan02 homograft tumor model was used to evaluate the in vivo efficacy of the p-toluenesulfonate salt of the compound of formula I and its combination with PD-1 antibody. After 31 days of observation, the treatment groups of PD-1 (10 mg / kg), p-toluenesulfonate salt of the compound of formula I + PD-1 (25 + 10 mg / kg), and p-toluenesulfonate salt of the compound of formula I + PD-1 (50 + 10 mg / kg) all showed significant tumor inhibition effects, with corresponding TGIs of 40.56%, 43.69%, and 60.31%, respectively. This indicates that the p-toluenesulfonate salt of the compound of formula I combined with PD-1 antibody can significantly inhibit the growth of mouse pancreatic cancer cell Pan02 homograft tumors in vivo.

[0173] During the experiment, the weight of the animals in each group increased steadily and there was no abnormality in their condition. That is, the test substance had no obvious toxicity to mice at the experimental dose.

[0174] All experimental operations strictly complied with standard experimental operating procedures and were carried out according to the experimental operations described in this study protocol.

[0175] Example 2: In vivo antitumor efficacy of the p-toluenesulfonate salt of the compound of formula I in a mouse H22 liver cancer model

[0176] Experimental Materials:

[0177] Animal preparation: Female BALB / c animals were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. and kept in an SPF animal room. The animals were allowed to adapt for at least one week before the experiment. Free access to food and water was allowed, the temperature was 20℃ to 25℃, the relative humidity was 40% to 70%, and the light hours were from 5 am to 5 pm every day.

[0178] Cell culture: H22 cells were cultured in RPMI 1640 medium supplemented with 10% FBS and 1% mycin-streptomycin in a 37°C, 5% CO2 incubator.

[0179] Cell inoculation: Collect cells and adjust the cell density to 2.0×10 7 / mL; mice were inoculated subcutaneously on the right back, with an inoculation volume of 0.05 mL per animal and an inoculation volume of 1×10 6 cells.

[0180] Preparation of p-toluenesulfonate salt of compound of formula I: The p-toluenesulfonate salt of compound of formula I is prepared in a solvent of 0.5% methylcellulose + 0.5% Tween-80, sterilized by high pressure, and stored at 4°C. The p-toluenesulfonate salt of compound of formula I (correction factor = 1.45) is prepared fresh before each administration, mixed by vortexing, and diluted with the solvent to a final concentration of 1.5 mg / mL, 5 mg / mL, and 10 mg / mL, as a clear solution; the oral administration volume is 10 mL / kg.

[0181] Preparation of reference compound LY2157299: LY2157299 solvent was 0.5% methylcellulose + 0.5% Tween-80, sterilized by high pressure, and stored at 4°C. LY2157299 (correction factor = 1.02) was freshly prepared before each administration. It was diluted with solvent to a final concentration of 5 mg / mL, and the oral administration volume was 10 mL / kg.

[0182] Experimental process:

[0183] Animal drug administration

[0184] When the tumor grows to 50-140 mm 3 (D6), 40 tumor-bearing mice were randomly divided into 5 groups, 8 mice in each group, respectively:

[0185] G1: vehicle control group (oral administration of p-toluenesulfonate vehicle of the compound of formula I);

[0186] G2: low-dose group of p-toluenesulfonate salt of the compound of formula I (15 mg / kg);

[0187] G3: medium dose group of p-toluenesulfonate salt of the compound of formula I (50 mg / kg);

[0188] G4: high dose of p-toluenesulfonate salt of the compound of formula I (100 mg / kg);

[0189] G5: LY2157299 administration group (50 mg / kg).

[0190] The p-toluenesulfonate salt of the compound of formula I and LY2157299 were both administered orally (po) twice a day (BID); the administration was continued for 15 days. The animals were weighed daily with an electronic balance, and the tumor volume was investigated with a vernier caliper three times a week. During the administration period, when the relative body weight change (RCBW) of the animal was less than -10%, the dosage was halved, and when it was less than <-15%, the administration was stopped until the RCBW returned to above -15%. The animals were clinically observed daily, and when the subcutaneous tumor volume exceeded 2000mm 3Animals were euthanized when they showed severe weight loss (RCBW<-20%) or other severe clinical symptoms. The dosing schedule is shown in Table 4.

[0191] Table 4 Dosage regimen

[0192]

[0193] At the end of the experiment (D21), all animals were euthanized, and the tumors were removed, embedded in paraffin, and prepared into FFPE blocks.

[0194] Efficacy evaluation method

[0195] The subcutaneous tumor volume was calculated using the following formula:

[0196] Tumor volume (TV) = (L×W2) / 2, where L is the long diameter of the tumor and W is the wide diameter of the tumor. The tumor volume is used to calculate the tumor growth inhibition rate (TGI), and the TGI shown in percentage is used to indicate the anti-tumor activity of the drug.

[0197] The TGI formula is as follows:

[0198] TGI (%) = [1-avT i-0 / avC i-0 )]×100; where avT i-0 is the average tumor volume of the dosing group on a specific day minus the average tumor volume of the dosing group on the day of grouping; where avC i-0 The mean tumor volume of the vehicle control group on a particular day was subtracted from the mean tumor volume of the vehicle control group on the day of grouping. Tumor volume data are shown as mean ± standard error (SEM).

[0199] The relative body weight change (RCBW) was calculated using the following formula: RCBW (%) = (BW i -BW0) / BW0×100; where BW i is the body weight of the animal on a particular day, and BW0 is the body weight of the animal on the day of grouping. Animal body weight (BW) and relative body weight change (RCBW) are also shown as mean ± standard error (SEM).

[0200] Statistical analysis methods

[0201] The tumor volumes of the groups were compared statistically using two-way ANOVA (Dunnett's multiple comparisons test). When P < 0.05, the difference was considered statistically significant. The tumor volumes were first logarithmically transformed to base 10 (animals with complete tumor regression were considered to have a tumor volume of 10 mm 3), and then two-way ANOVA was performed.

[0202] The tumor weights between the groups were statistically compared using one-way ANOVA (Dunnett's multiple comparisons test). When P < 0.05, the difference was considered statistically significant. The tumor weights were first logarithmically transformed to base 10 (animals with complete tumor regression were considered to have tumor weight = 0.01 g) and then subjected to one-way ANOVA.

[0203] All graphs and statistical analyses were performed using GraphPad Prism 6.00 software (GraphPad Software).

[0204] Experimental results:

[0205] H22 tumors grow well subcutaneously in BALB / c mice, and the antitumor activity of the p-toluenesulfonate salt of the compound of Formula I was observed in this experiment. At the end of the experiment (D21), the tumor volumes of the vehicle control group, the low-dose p-toluenesulfonate salt group of the compound of Formula I (15 mg / kg), the medium-dose p-toluenesulfonate salt group of the compound of Formula I (50 mg / kg), the high-dose p-toluenesulfonate salt group of the compound of Formula I (100 mg / kg), and the LY2157299 administration group (50 mg / kg) were 2267.17±223.44 mm, respectively. 3 , 930.22±291.18mm 3 , 1000.58±191.78mm 3 , 1061.15±226.94mm 3 and 1377.01±698.94mm 3 .

[0206] At the end of the experiment (D21), the tumor growth inhibition rates (TGI) of the corresponding administration groups were 61.6%, 58.3%, 55.6%, and 40.9%, respectively; the low-dose group (15 mg / kg) and the medium-dose group (50 mg / kg) of the p-toluenesulfonate salt of the compound of formula I were able to significantly inhibit the growth of H22 tumors, and the efficacy was better than that of the LY2157299 administration group (50 mg / kg). Tumor growth data are shown in Figure 3 , Figure 4 and Table 5.

[0207] At the end of the experiment (D21), the average animal body weights of the vehicle control group, the low-dose group of the p-toluenesulfonate of the compound of Formula I (15 mg / kg), the medium-dose group of the p-toluenesulfonate of the compound of Formula I (50 mg / kg), the high-dose group of the p-toluenesulfonate of the compound of Formula I (100 mg / kg) and the LY2157299 administration group (50 mg / kg) were 24.35±0.35 g, 22.14±0.73 g, 23.14±0.30 g, 24.44±0.67 g, 22.85±0.88 g, respectively, and the average relative body weight changes were 15.7±2.1%, 7.7±4.3%, 7.7±0.6%, 17.4±3.6%, 8.1±5.7%, respectively; within the dosage and cycle, the weight of the animals increased steadily, and no abnormal conditions of the animals were observed.

[0208] Table 5 Summary of in vivo efficacy evaluation of the p-toluenesulfonate salt of the compound of formula I in the mouse liver cancer H22 model

[0209]

[0210]

[0211] Note:

[0212] a. Mean ± SEM, n = 8.

[0213] b. Tumor (volume) growth inhibition rate TGI (%) = [(1-(average tumor volume of a treatment group at the end of drug administration - average tumor volume of the treatment group at the beginning of drug administration)) / (average tumor volume of the solvent control group at the end of treatment - average tumor volume of the solvent control group at the beginning of treatment)] × 100.

[0214] c. Based on data from day 21 after vaccination.

[0215] d. Tumor disappearance is the number of mice with tumor disappearance / number of animals in each group.

[0216] Experimental conclusion:

[0217] In the mouse H22 liver cancer model, monotherapy with different doses of p-toluenesulfonate salt of the compound of formula I (15 mg / kg, 50 mg / kg and 100 mg / kg) can inhibit tumor growth; compared with the vehicle control group, the p-toluenesulfonate salt administration groups of 15 mg / kg and 50 mg / kg of the compound of formula I have significant differences; within the dosage range used, no dose-dependent pharmacodynamic relationship of p-toluenesulfonate salt of the compound of formula I was observed. Compared with the vehicle control group, monotherapy with 50 mg / kg of the reference compound LY2157299 can partially inhibit tumor growth. In the H22 model, the efficacy of monotherapy with different doses of p-toluenesulfonate salt of the compound of formula I (15 mg / kg, 50 mg / kg and 100 mg / kg) is better than that of 50 mg / kg of the reference compound LY2157299 ( Figure 3 Within the dosage and cycle, monotherapy with different doses of the p-toluenesulfonate salt of the compound of Formula I and monotherapy with LY2157299 had no obvious side effects on the animals, and the weight of the animals increased steadily ( Figure 4 ).

[0218] The foregoing description of specific exemplary embodiments of the present invention is for the purpose of illustration and demonstration. These descriptions are not intended to limit the present invention to the precise form disclosed, and it is clear that many changes and variations can be made based on the above teachings. The purpose of selecting and describing the exemplary embodiments is to explain the specific principles of the present invention and its practical application, so that those skilled in the art can realize and utilize various different exemplary embodiments of the present invention and various different selections and changes. The scope of the present invention is intended to be limited by the claims and their equivalents.

Claims

1. A pharmaceutical combination or kit comprising a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of an immunotherapeutic agent; The immunosuppressant is a therapeutic agent or a combination thereof against the following targets: PD-1, PD-L1, CTLA-4, PD-L2, LAG3, TIM3, 2B4, A2aR, B7H1, B7H3, B7H4, BTLA, CD2, CD27, CD28, CD30, CD40, CD70, CD80, CD86, CD137, CD160, CD226, CD276, DR3, GAL9, GITR, HAVCR2, HVEM, IDO1, IDO2, ICOS, KIR, LAIR1, LIGHT, MARCO, PS, OX-40, SLAM, TIGHT, VISTA or VTCN1.

2. The pharmaceutical combination or kit according to claim 1, wherein: The immunotherapeutic agent is an anti-PD-1 antibody or an antigen-binding fragment thereof; Preferably, the immunotherapeutic agent is selected from Galunisertib, Teplizumab, AMP-224, Zimberelimab, Sintilimab, Tislelizumab, Pidolizumab, Pembrolizumab, LZM-009, Nivolumab, Sintilimab, Cimiprilimab, Tislelizumab, Dotalimumab, BI-754091 or Spartalizumab.

3. The pharmaceutical combination or kit according to claim 1 or 2, wherein: The pharmaceutically acceptable salt of the compound of formula I is selected from benzenesulfonate, p-toluenesulfonate, methanesulfonate, ethanesulfonate, hydrochloride or sulfate.

4. The pharmaceutical combination or kit according to any one of claims 1 to 3, wherein: The drug combination or drug kit is used to treat tumors; Preferably, the tumor is an advanced or metastatic tumor.

5. The pharmaceutical combination or kit according to claim 4, wherein: The tumor is a solid tumor; Preferably, the tumor is selected from pelvic paraganglioma, small cell lung cancer, non-small cell lung cancer, nasopharyngeal carcinoma, gastric cancer, intestinal cancer, colon cancer, breast cancer, cervical cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumors, ovarian tumors, peritoneal tumors, melanoma, glioma, glioblastoma, hepatocellular carcinoma, papillary nephroma, head and neck squamous cell carcinoma, esophageal squamous cell carcinoma, endometrial cancer, synovial sarcoma, thymic carcinoma, bile duct cancer, thyroid cancer, bile duct cancer / gallbladder cancer, colorectal cancer, urothelial carcinoma, esophageal cancer, gastric and esophageal junction adenocarcinoma, pleural mesothelioma or Hodgkin's lymphoma.

6. Use of the drug combination or drug kit according to any one of claims 1 to 5 in the preparation of anti-tumor drugs.

7. The use according to claim 6, wherein The compound of formula I or the immunotherapeutic agent is continuously or intermittently administered to the subject; Preferably, the continuous administration of the compound of formula I or the immunotherapeutic agent is interrupted by one or more drug holidays.

8. The pharmaceutical combination or kit according to any one of claims 1 to 5, wherein: The kit comprises a container, a pharmaceutical composition contained in the container, and a package insert containing instructions for the pharmaceutical composition.

9. Use of a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of an anti-tumor drug.

10. The use according to claim 9, wherein The pharmaceutically acceptable salt of the compound of formula I is selected from benzenesulfonate, p-toluenesulfonate, methanesulfonate, ethanesulfonate, hydrochloride or sulfate; Preferably, the tumor is selected from pelvic paraganglioma, small cell lung cancer, non-small cell lung cancer, nasopharyngeal carcinoma, gastric cancer, intestinal cancer, colon cancer, cervical cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer, oral cancer, prostate cancer, bone cancer, kidney cancer, ovarian cancer, bladder cancer, liver cancer, fallopian tube tumors, ovarian tumors, peritoneal tumors, melanoma, glioma, glioblastoma, hepatocellular carcinoma, papillary nephroma, head and neck squamous cell carcinoma, esophageal squamous cell carcinoma, endometrial cancer, synovial sarcoma, thymic carcinoma, bile duct cancer, thyroid cancer, bile duct cancer / gallbladder cancer, colorectal cancer, urothelial carcinoma, esophageal cancer, gastric and esophageal junction adenocarcinoma, pleural mesothelioma or Hodgkin's lymphoma.

Images