Therapeutic for cancer refractory to immune checkpoint inhibitor

Abstract

Provided is a compound that exhibits anti-cancer activity based on CHK1 inhibition. The present disclosure is a result of the discovery that compounds represented by formula (1) or pharmacologically acceptable salts thereof have excellent antitumor activity due to strong inhibitory activity against CHK1. (In the formula, R1, R2, L, V, W, and Q are as defined in the description, and X, Y, and Z each independently represent CR8 or a nitrogen atom. X, Y, and Z are not simultaneously CR8, and R8 is as defined in the description.)

Description

Drugs for treating cancers that are resistant to immune checkpoint inhibitors

[0001] The present disclosure relates to a CHK1 inhibitor, a liposome encapsulating a CHK1 inhibitor, or a combination of these with an immune checkpoint inhibitor, as a therapeutic agent for cancer that exhibits resistance to treatment with an immune checkpoint inhibitor.

[0002] CHK1 is a serine / threonine protein kinase downstream of ATR in the DNA damage checkpoint signaling pathway during the cell cycle. In mammalian cells, CHK1 is phosphorylated in an ATR-dependent manner in response to DNA damage caused by ionizing radiation or DNA replication stress resulting from excessive cell proliferation or genomic instability in cancer cells (Non-Patent Documents 1-5). This phosphorylation activates CHK1, causing CHK1 to phosphorylate CDC25A, inhibiting CDC25A-mediated dephosphorylation of cyclin E / CDK2 and halting progression from the S phase. CHK1 also phosphorylates CDC25C, inhibiting CDC25C-mediated dephosphorylation of cyclin B / CDK1, halting progression from the G2M phase. In both cases, regulation of CDK activity induces cell cycle arrest, preventing cell division in the presence of DNA damage and promoting DNA damage repair. CHK1 inhibition suppresses the checkpoint function of DNA damage in the cell cycle in cancer cells, causing DNA repair failure and uncontrollable DNA synthesis in the presence of DNA damage, resulting in DNA fragmentation, replication catastrophe, and cell death (Patent Documents 1 and 2).

[0003] Since their launch, immune checkpoint inhibitors have established themselves as the fourth line of cancer treatment, following surgery, radiation therapy, and chemotherapy. These immune checkpoint inhibitors are currently approved for a wide range of cancer types, including melanoma. While their efficacy has been reported to be characterized by long-term survival (durable clinical response), this effect is only observed in a small proportion of patients. Therefore, improvements in response rates are needed, such as the 13.3% overall response rate of nivolumab in head and neck squamous cell carcinoma (Non-Patent Document 7).

[0004] Various inhibitors of CHK1 have been reported (Patent Documents 1 to 3).

[0005] International Publication No. 2010 / 077758 International Publication No. 2012 / 064548 International Publication No. 2017 / 132928

[0006] Dai Y and Grant S, Clin Cancer Res, 16(2):376-383 (2010)Benada J and Macurek L, Biomolecules, 5:1912-1937 (2015)King C, Mol Cancer Ther, 14(9):2004-2013 (2015)Dent P, Expert Opinion on Investigational drugs, 28(12):1095-1100 (2019)Evangelisti G. et al. Expert Opinion on Investigational Drugs, 29(8):779-792 (2020)David H. et al. J Clin Oncol, 34:1764-1771 (2016)Raffaele A. et al. Expert Opinion on Biological Therapy, 19:3, 169-171 (2019)

[0007] The present disclosure provides agents for treating cancers that are resistant to immune checkpoint inhibitors or for preventing recurrence of cancers that have gone into remission.

[0008] As a result of extensive research, the present inventors have found that a CHK1 inhibitor or a liposome encapsulating a CHK1 inhibitor enhances cytocidal effects and antigen presentation by continuously exposing the drug to cancer tissue, and that M2 macrophages are reduced, creating an intratumor environment (inflammatory microenvironment) that attracts CD8-positive T cells. They have also found that the agent exhibits excellent antitumor effects against cancers that are resistant to immune checkpoint inhibitor therapy. Furthermore, they have found that combining the inhibitor or liposome with an immune checkpoint inhibitor exhibits even more excellent antitumor effects. Furthermore, they have found that combining the inhibitor or liposome with an immune checkpoint inhibitor increases CD8-positive T cells with memory function, resulting in tumor rejection or tumor growth suppression upon secondary transplantation, thereby demonstrating a preventive effect against the recurrence of cancers in remission. This discovery has led to the completion of the present disclosure.

[0009] That is, the present disclosure is as follows. [Item 1] A therapeutic and / or prophylactic agent for cancer patients showing resistance to immune checkpoint inhibitors, comprising as an active ingredient a CHK1 inhibitor or a liposome encapsulating a CHK1 inhibitor. [Item 2] The therapeutic and / or prophylactic agent according to Item 1, characterized in that the CHK1 inhibitor and an immune checkpoint inhibitor are used in combination. [Item 3] The therapeutic and / or prophylactic agent according to Item 2, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously. [Item 4] The therapeutic and / or prophylactic agent according to any one of Items 2 or 3, characterized in that the immune checkpoint inhibitor is administered after the CHK1 inhibitor is administered. [Item 5] The therapeutic and / or prophylactic agent according to any one of Items 2 to 4, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously, and then the anti-PD-1 antibody is administered. [Item 6] The therapeutic and / or prophylactic agent according to any one of Items 2 to 4, wherein the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously, and then the anti-PD-1 antibody is administered at least three days later. [Item 7] The therapeutic and / or prophylactic agent according to any one of Items 2 to 6, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody. [Item 8] The therapeutic and / or prophylactic agent according to any one of Items 2 to 6, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody. [Item 9] The therapeutic and / or prophylactic agent according to any one of Items 2 to 6, wherein the anti-PD-1 antibody is pembrolizumab, nivolumab, spartalizumab, or cemiplimab. [Item 10] The therapeutic and / or prophylactic agent according to any one of Items 1 to 9, wherein the number of CD3-positive cells in tumor tissue of the cancer patient is reduced compared to tumor tissue of a patient responsive to the anti-PD-1 antibody. [Item 11] The therapeutic and / or prophylactic agent according to any one of Items 1 to 10, wherein the number of CD8-positive cells in tumor tissue of the cancer patient is reduced compared to tumor tissue of a patient responsive to an anti-PD-1 antibody.[Item 12] The therapeutic and / or preventive agent according to any one of Items 1 to 11, wherein the number of CD8-positive cells in the tumor tissue of the cancer patient is 0.135 or less. [Item 13] The therapeutic and / or preventive agent according to any one of Items 1 to 12, wherein the M1-M2 ratio of macrophages in the tumor tissue of the cancer patient is 21 or less. [Item 14] The therapeutic and / or preventive agent according to any one of Items 1 to 13, wherein the number of CD206-positive cells in the tumor tissue of the cancer patient is increased compared to tumor tissue of a patient responsive to an anti-PD-1 antibody. [Item 15] The therapeutic and / or preventive agent according to any one of Items 1 to 14, wherein the number of IAd-positive cells in the tumor tissue of the cancer patient is decreased compared to tumor tissue of a patient responsive to an anti-PD-1 antibody. [Item 16] The therapeutic and / or prophylactic agent according to any one of Items 1 to 15, wherein the number of CD62L-negative CD44-positive cells in the lymph nodes of the cancer patient is reduced compared to the lymph nodes of patients responsive to an anti-PD-1 antibody. [Item 17] The therapeutic and / or prophylactic agent according to any one of Items 1 to 16, wherein PD-L1 expression in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 18] The therapeutic and / or prophylactic agent according to any one of Items 1 to 17, wherein VEGFa expression in the tumor tissue of the cancer patient is increased compared to the tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 19] The therapeutic and / or prophylactic agent according to any one of Items 1 to 17, wherein Granzyme B expression in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 20] The therapeutic and / or prophylactic agent according to any one of Items 1 to 17, wherein Interferon γ expression in the tumor tissue of the cancer patient is reduced compared to tumor tissue of a patient responsive to an anti-PD-1 antibody. [Item 21] The therapeutic and / or prophylactic agent according to any one of Items 1 to 17, wherein H2abl expression in the tumor tissue of the cancer patient is reduced compared to tumor tissue of a patient responsive to an anti-PD-1 antibody.[Item 22] The therapeutic and / or prophylactic agent according to any one of Items 1 to 17, wherein CD80 expression in the tumor tissue of the cancer patient is reduced compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 23] The therapeutic and / or prophylactic agent according to any one of Items 1 to 17, wherein Mrc1 expression in the tumor tissue of the cancer patient is increased compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 24] The therapeutic and / or prophylactic agent according to any one of Items 1 to 17, wherein Arginase 1 expression in the tumor tissue of the cancer patient is increased compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 25] The therapeutic and / or prophylactic agent according to any one of Items 1 to 17, wherein DNA replication stress in the tumor tissue of the cancer patient is reduced compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 26] A therapeutic and / or prophylactic agent for preventing the recurrence of cancer that has gone into remission, comprising as an active ingredient a CHK1 inhibitor or a liposome encapsulating a CHK1 inhibitor. [Item 27] ​​The therapeutic and / or prophylactic agent according to Item 26, characterized in that the CHK1 inhibitor is used in combination with an immune checkpoint inhibitor. [Item 28] The therapeutic and / or prophylactic agent according to either Item 26 or 27, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody. [Item 29] The therapeutic and / or prophylactic agent according to either Item 26 or 27, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody. [Item 30] The therapeutic and / or prophylactic agent according to either Item 26 or 27, characterized in that the anti-PD-1 antibody is pembrolizumab, nivolumab, spartalizumab, or cemiplimab.[Item 31] The cancer is selected from the group consisting of squamous cell carcinoma, melanoma, acute leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, myelodysplastic syndrome, polycythemia vera, malignant lymphoma, plasma cell neoplasm, multiple myeloma, brain tumor, head and neck cancer, head and neck squamous cell carcinoma, esophageal cancer, thyroid cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, thymoma / thymic carcinoma, breast cancer, triple-negative breast cancer, gastric cancer, gallbladder / bile duct cancer, liver cancer, hepatocellular carcinoma, pancreatic cancer, colon cancer, rectal cancer, MSI-high colorectal cancer, anal cancer, and gastrointestinal cancer. Item 31. The therapeutic and / or preventive agent according to any one of Items 26 to 30, wherein the cancer is at least one type selected from the group consisting of tubular stromal tumor, chorioepithelial carcinoma, endometrial cancer, cervical cancer, ovarian cancer, platinum-resistant ovarian cancer, PARP inhibitor-resistant ovarian cancer, bladder cancer, urothelial cancer, kidney cancer, renal cell carcinoma, prostate cancer, testicular tumor, testicular germ cell tumor, ovarian germ cell tumor, Wilms' tumor, malignant melanoma, neuroblastoma, osteosarcoma, Ewing's sarcoma, chondrosarcoma, soft tissue sarcoma, and skin cancer. [Item 32] The therapeutic and / or preventive agent according to any one of Items 26 to 30, wherein the cancer is at least one type of cancer selected from the group consisting of melanoma, squamous cell carcinoma, non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, urothelial cancer, non-muscle-invasive bladder cancer, kidney cancer, hepatocellular carcinoma, MSI-high colorectal cancer, esophageal cancer, colon cancer, rectal cancer, breast cancer, and endometrial cancer. [Item 33] The therapeutic and / or preventive agent according to any one of Items 26 to 30, wherein the cancer is at least one type of cancer selected from the group consisting of head and neck cancer, squamous cell carcinoma, non-small cell lung cancer, anal cancer, breast cancer, triple-negative breast cancer, ovarian cancer, platinum-resistant ovarian cancer, and endometrial cancer. [Item 34] The therapeutic and / or preventive agent according to any one of Items 26 to 30, wherein the cancer is a solid cancer with a high tumor mutation burden (Tumor Mutation Burden-High; TMB-High). [Item 35] The therapeutic and / or preventive agent according to any one of Items 1 to 34, wherein the liposome further comprises a phospholipid.[Item 36] The therapeutic and / or prophylactic agent according to Item 35, wherein the phospholipid is one or a combination of two or more selected from the group consisting of phosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, soybean lecithin, egg yolk lecithin, hydrogenated egg yolk lecithin, and hydrogenated soybean lecithin. [Item 37] The therapeutic and / or prophylactic agent according to any one of Items 1 to 36, wherein the liposome further comprises a sterol. [Item 38] The therapeutic and / or prophylactic agent according to any one of Items 1 to 37, wherein the sterol is cholesterol. [Item 39] The therapeutic and / or prophylactic agent according to any one of Items 1 to 38, wherein the liposome further comprises a polymer-modified lipid. [Item 40] The therapeutic and / or prophylactic agent according to Item 39, wherein the polymer moiety of the polymer-modified lipid is polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, methoxypolyethylene glycol, methoxypolypropylene glycol, methoxypolyvinyl alcohol, methoxypolyvinylpyrrolidone, ethoxypolyethylene glycol, ethoxypolypropylene glycol, ethoxypolyvinyl alcohol, ethoxypolyvinylpyrrolidone, propoxypolyethylene glycol, propoxypolypropylene glycol, propoxypolyvinyl alcohol, or propoxypolyvinylpyrrolidone. [Item 41] The therapeutic and / or prophylactic agent according to Item 39 or 40, wherein the lipid moiety of the polymer-modified lipid is phosphatidylethanolamine or diacylglycerol. [Item 42] The therapeutic and / or prophylactic agent according to any one of Items 1 to 41, wherein the liposome encapsulating the CHK1 inhibitor comprises: (1) 40 to 70 mol % of phospholipid, (2) 30 to 50 mol % of cholesterol, and (3) 1 to 10 mol % of a polymer-modified lipid. [Item 43] The therapeutic and / or prophylactic agent according to any one of Items 1 to 42, wherein the liposome further comprises an additive selected from the group consisting of inorganic acids, inorganic acid salts, organic acids, organic acid salts, sugars, buffers, antioxidants, and polymers.[Item 1A] Use of a CHK1 inhibitor or a liposome encapsulating a CHK1 inhibitor for the manufacture of a therapeutic and / or prophylactic agent for cancer patients showing resistance to immune checkpoint inhibitors. [Item 2A] The use according to Item 1A, characterized in that the CHK1 inhibitor and an immune checkpoint inhibitor are used in combination. [Item 3A] The use according to Item 2A, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously. [Item 4A] The use according to any one of Item 2A or 3A, characterized in that the immune checkpoint inhibitor is administered after the CHK1 inhibitor. [Item 5A] The use according to any one of Item 2A to 4A, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously, and then the anti-PD-1 antibody is administered. [Item 6A] The use according to any one of Item 2A to 4A, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously, and then the anti-PD-1 antibody is administered three days or more later. [Item 7A] The use of any one of Items 2A to 6A, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody. [Item 8A] The use of any one of Items 2A to 6A, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody. [Item 9A] The use of any one of Items 2A to 6A, wherein the anti-PD-1 antibody is pembrolizumab, nivolumab, spartalizumab, or cemiplimab. [Item 10A] The use of any one of Items 1A to 9A, wherein the number of CD3-positive cells in the tumor tissue of the cancer patient is reduced compared to tumor tissue of a patient responsive to the anti-PD-1 antibody. [Item 11A] The use of any one of Items 1A to 10A, wherein the number of CD8-positive cells in the tumor tissue of the cancer patient is reduced compared to tumor tissue of a patient responsive to the anti-PD-1 antibody. [Item 12A] The use according to any one of Items 1A to 11A, wherein the number of CD8-positive cells in the tumor tissue of the cancer patient is 0.135 or less.[Item 13A] The use of any one of Items 1A to 12A, wherein the M1-M2 ratio of macrophages in the tumor tissue of the cancer patient is 21 or less. [Item 14A] The use of any one of Items 1A to 13A, wherein the number of CD206-positive cells in the tumor tissue of the cancer patient is increased compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 15A] The use of any one of Items 1A to 14A, wherein the number of IAd-positive cells in the tumor tissue of the cancer patient is decreased compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 16A] The use of any one of Items 1A to 15A, wherein the number of CD62L-negative CD44-positive cells in the lymph nodes of the cancer patient is decreased compared to lymph nodes from patients responsive to an anti-PD-1 antibody. [Item 17A] The use of any one of Items 1A to 16A, wherein PD-L1 expression in the tumor tissue of the cancer patient is decreased compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 18A] The use of any one of Items 1A to 17A, wherein VEGFa expression in the tumor tissue of the cancer patient is increased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 19A] The use of any one of Items 1A to 17A, wherein Granzyme B expression in the tumor tissue of the cancer patient is decreased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 20A] The use of any one of Items 1A to 17A, wherein Interferon γ expression in the tumor tissue of the cancer patient is decreased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 21A] The use of any one of Items 1A to 17A, wherein H2abl expression in the tumor tissue of the cancer patient is decreased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 22A] The use of any one of Items 1A to 17A, wherein CD80 expression in tumor tissue of the cancer patient is reduced compared to tumor tissue of a patient responsive to an anti-PD-1 antibody.[Item 23A] The use according to any one of Items 1A to 17A, characterized in that Mrc1 expression in the tumor tissue of the cancer patient is increased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 24A] The use according to any one of Items 1A to 17A, characterized in that Arginase 1 expression in the tumor tissue of the cancer patient is increased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 25A] The use according to any one of Items 1A to 17A, characterized in that DNA replication stress in the tumor tissue of the cancer patient is reduced compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 26A] Use of a CHK1 inhibitor or a liposome encapsulating a CHK1 inhibitor for the manufacture of a therapeutic and / or prophylactic agent for preventing the recurrence of cancer that has gone into remission. [Item 27A] The use according to Item 26A, characterized in that the CHK1 inhibitor is used in combination with an immune checkpoint inhibitor. [Item 28A] The use of either one of Items 26A or 27A, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody. [Item 29A] The use of either one of Items 26A or 27A, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody. [Item 30A] The use of either one of Items 26A or 27A, wherein the anti-PD-1 antibody is pembrolizumab, nivolumab, spartalizumab, or cemiplimab.[Item 31A] The cancer is selected from the group consisting of squamous cell carcinoma, melanoma, acute leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, myelodysplastic syndrome, polycythemia vera, malignant lymphoma, plasma cell neoplasm, multiple myeloma, brain tumor, head and neck cancer, head and neck squamous cell carcinoma, esophageal cancer, thyroid cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, thymoma / thymic carcinoma, breast cancer, triple-negative breast cancer, gastric cancer, gallbladder / bile duct cancer, liver cancer, hepatocellular carcinoma, pancreatic cancer, colon cancer, rectal cancer, MSI-high colorectal cancer, and anal cancer. The use of any one of Items 26A to 30A, wherein the cancer is at least one type of cancer selected from the group consisting of cancer, gastrointestinal stromal tumor, chorioepithelial carcinoma, endometrial cancer, cervical cancer, ovarian cancer, platinum-resistant ovarian cancer, PARP inhibitor-resistant ovarian cancer, bladder cancer, urothelial cancer, kidney cancer, renal cell carcinoma, prostate cancer, testicular tumor, testicular germ cell tumor, ovarian germ cell tumor, Wilms' tumor, malignant melanoma, neuroblastoma, osteosarcoma, Ewing's sarcoma, chondrosarcoma, soft tissue sarcoma, and skin cancer. [Item 32A] The use of any one of Items 26A to 30A, wherein the cancer is melanoma, non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, urothelial cancer, non-muscle-invasive bladder cancer, renal cancer, hepatocellular carcinoma, MSI-high colorectal cancer, esophageal cancer, colorectal cancer, breast cancer, or endometrial cancer. [Item 33A] The use of any one of Items 26A to 30A, wherein the cancer is at least one type of cancer selected from the group consisting of head and neck cancer, squamous cell carcinoma, non-small cell lung cancer, anal cancer, breast cancer, triple-negative breast cancer, ovarian cancer, platinum-resistant ovarian cancer, and endometrial cancer. [Item 34A] The use of any one of Items 26A to 30A, wherein the cancer is a solid cancer with a high tumor mutation burden (Tumor Mutation Burden-High; TMB-High). [Item 35A] The use of any one of Items 1A to 34A, wherein the liposome further comprises a phospholipid.[Item 36A] The use according to Item 35A, wherein the phospholipid is one or a combination of two or more selected from the group consisting of phosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, soybean lecithin, egg yolk lecithin, hydrogenated egg yolk lecithin, and hydrogenated soybean lecithin. [Item 37A] The use according to any one of Items 1A to 36A, wherein the liposome further comprises a sterol. [Item 38A] The use according to any one of Items 1A to 37A, wherein the sterol is cholesterol. [Item 39A] The use according to any one of Items 1A to 38A, wherein the liposome further comprises a polymer-modified lipid. [Item 40A] The use according to Item 39A, wherein the polymer moiety of the polymer-modified lipid is polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, methoxypolyethylene glycol, methoxypolypropylene glycol, methoxypolyvinyl alcohol, methoxypolyvinylpyrrolidone, ethoxypolyethylene glycol, ethoxypolypropylene glycol, ethoxypolyvinyl alcohol, ethoxypolyvinylpyrrolidone, propoxypolyethylene glycol, propoxypolypropylene glycol, propoxypolyvinyl alcohol, or propoxypolyvinylpyrrolidone. [Item 41A] The use according to Item 39A or 40A, wherein the lipid moiety of the polymer-modified lipid is phosphatidylethanolamine or diacylglycerol. [Item 42A] The use according to any one of Items 1A to 41A, wherein the liposome encapsulating the CHK1 inhibitor comprises: (1) 40 to 70 mol % of phospholipid, (2) 30 to 50 mol % of cholesterol, and (3) 1 to 10 mol % of polymer-modified lipid. [Item 43A] The use according to any one of Items 1A to 42A, wherein the liposome further comprises an additive selected from the group consisting of inorganic acids, inorganic acid salts, organic acids, organic acid salts, sugars, buffers, antioxidants, and polymers.[Item 1B] A therapeutic and / or prophylactic agent containing, as an active ingredient, a CHK1 inhibitor or a liposome encapsulating a CHK1 inhibitor, for use in the treatment and / or prevention of cancer patients exhibiting resistance to immune checkpoint inhibitors. [Item 2B] The therapeutic and / or prophylactic agent according to Item 1B, characterized in that the CHK1 inhibitor and an immune checkpoint inhibitor are used in combination. [Item 3B] The therapeutic and / or prophylactic agent according to Item 2B, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously. [Item 4B] The therapeutic and / or prophylactic agent according to any one of Item 2B or 3B, characterized in that the immune checkpoint inhibitor is administered after the CHK1 inhibitor is administered. [Item 5B] The therapeutic and / or prophylactic agent according to any one of Item 2B to 4B, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously, and then the anti-PD-1 antibody is administered. [Item 6B] The therapeutic and / or prophylactic agent according to any one of Items 2B to 4B, wherein the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously, and then the anti-PD-1 antibody is administered at least three days later. [Item 7B] The therapeutic and / or prophylactic agent according to any one of Items 2B to 6B, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody. [Item 8B] The therapeutic and / or prophylactic agent according to any one of Items 2B to 6B, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody. [Item 9B] The therapeutic and / or prophylactic agent according to any one of Items 2B to 6B, wherein the anti-PD-1 antibody is pembrolizumab, nivolumab, spartalizumab, or cemiplimab. [Item 10B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 9B, wherein the number of CD3-positive cells in the tumor tissue of the cancer patient is reduced compared to that in tumor tissue of a patient responsive to an anti-PD-1 antibody. [Item 11B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 10B, wherein the number of CD8-positive cells in the tumor tissue of the cancer patient is reduced compared to that in tumor tissue of a patient responsive to an anti-PD-1 antibody.[Item 12B] The therapeutic and / or preventive agent according to any one of Items 1B to 11B, wherein the number of CD8-positive cells in the tumor tissue of the cancer patient is 0.135 or less. [Item 13B] The therapeutic and / or preventive agent according to any one of Items 1B to 12B, wherein the M1-M2 ratio of macrophages in the tumor tissue of the cancer patient is 21 or less. [Item 14B] The therapeutic and / or preventive agent according to any one of Items 1B to 13B, wherein the number of CD206-positive cells in the tumor tissue of the cancer patient is increased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 15B] The therapeutic and / or preventive agent according to any one of Items 1B to 14B, wherein the number of IAd-positive cells in the tumor tissue of the cancer patient is decreased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 16B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 15B, wherein the number of CD62L-negative CD44-positive cells in the lymph nodes of the cancer patient is reduced compared to the lymph nodes of patients responsive to an anti-PD-1 antibody. [Item 17B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 16B, wherein PD-L1 expression in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 18B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 17B, wherein VEGFa expression in the tumor tissue of the cancer patient is increased compared to the tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 19B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 17B, wherein Granzyme B expression in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 20B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 17B, wherein Interferon γ expression in tumor tissue of the cancer patient is reduced compared to tumor tissue of a patient responsive to an anti-PD-1 antibody.[Item 21B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 17B, characterized in that H2abl expression in the tumor tissue of the cancer patient is reduced compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 22B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 17B, characterized in that CD80 expression in the tumor tissue of the cancer patient is reduced compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 23B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 17B, characterized in that Mrc1 expression in the tumor tissue of the cancer patient is increased compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 24B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 17B, characterized in that Arginase 1 expression in the tumor tissue of the cancer patient is increased compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 25B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 17B, characterized in that DNA replication stress in the tumor tissue of the cancer patient is reduced compared to tumor tissue of a patient responsive to an anti-PD-1 antibody. [Item 26B] A therapeutic and / or prophylactic agent containing, as an active ingredient, a CHK1 inhibitor or a liposome encapsulating a CHK1 inhibitor, for use in preventing recurrence of cancer that has gone into remission. [Item 27B] The therapeutic and / or prophylactic agent according to Item 26B, characterized in that the CHK1 inhibitor is used in combination with an immune checkpoint inhibitor. [Item 28B] The therapeutic and / or prophylactic agent according to any one of Items 26B or 27B, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody. [Item 29B] The therapeutic and / or prophylactic agent according to any one of Items 26B or 27B, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody. [Item 30B] The therapeutic and / or prophylactic agent according to any one of Items 26B and 27B, wherein the anti-PD-1 antibody is pembrolizumab, nivolumab, spartalizumab, or cemiplimab.[Item 31B] The cancer is selected from the group consisting of squamous cell carcinoma, melanoma, acute leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, myelodysplastic syndrome, polycythemia vera, malignant lymphoma, plasma cell neoplasm, multiple myeloma, brain tumor, head and neck cancer, head and neck squamous cell carcinoma, esophageal cancer, thyroid cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, thymoma / thymic carcinoma, breast cancer, triple-negative breast cancer, gastric cancer, gallbladder / bile duct cancer, liver cancer, hepatocellular carcinoma, pancreatic cancer, colon cancer, rectal cancer, MSI-high colorectal cancer, anal cancer, and gastrointestinal cancer. The therapeutic and / or prophylactic agent according to any one of Items 26B to 30B, wherein the cancer is at least one type selected from the group consisting of endothelial stromal tumor, chorioepithelial carcinoma, endometrial cancer, cervical cancer, ovarian cancer, platinum-resistant ovarian cancer, PARP inhibitor-resistant ovarian cancer, bladder cancer, urothelial cancer, kidney cancer, renal cell carcinoma, prostate cancer, testicular tumor, testicular germ cell tumor, ovarian germ cell tumor, Wilms' tumor, malignant melanoma, neuroblastoma, osteosarcoma, Ewing's sarcoma, chondrosarcoma, soft tissue sarcoma, and skin cancer. [Item 32B] The therapeutic and / or preventive agent according to any one of Items 26B to 30B, wherein the cancer is melanoma, non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, urothelial cancer, non-muscle-invasive bladder cancer, kidney cancer, hepatocellular carcinoma, MSI-high colorectal cancer, esophageal cancer, colorectal cancer, breast cancer, or endometrial cancer. [Item 33B] The therapeutic and / or preventive agent according to any one of Items 26B to 30B, wherein the cancer is at least one type of cancer selected from the group consisting of head and neck cancer, squamous cell carcinoma, non-small cell lung cancer, anal cancer, breast cancer, triple-negative breast cancer, ovarian cancer, platinum-resistant ovarian cancer, or endometrial cancer. [Item 34B] The therapeutic and / or preventive agent according to any one of Items 26B to 30B, wherein the cancer is a solid cancer with a high tumor mutation burden (Tumor Mutation Burden-High; TMB-High). [Item 35B] The therapeutic and / or preventive agent according to any one of Items 1B to 34B, wherein the liposome further comprises a phospholipid.[Item 36B] The therapeutic and / or prophylactic agent according to Item 35B, wherein the phospholipid is one or a combination of two or more selected from the group consisting of phosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, soybean lecithin, egg yolk lecithin, hydrogenated egg yolk lecithin, and hydrogenated soybean lecithin. [Item 37B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 36B, wherein the liposome further comprises a sterol. [Item 38B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 37B, wherein the sterol is cholesterol. [Item 39B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 38B, wherein the liposome further comprises a polymer-modified lipid. [Item 40B] The therapeutic and / or prophylactic agent according to Item 39B, wherein the polymer moiety of the polymer-modified lipid is polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, methoxypolyethylene glycol, methoxypolypropylene glycol, methoxypolyvinyl alcohol, methoxypolyvinylpyrrolidone, ethoxypolyethylene glycol, ethoxypolypropylene glycol, ethoxypolyvinyl alcohol, ethoxypolyvinylpyrrolidone, propoxypolyethylene glycol, propoxypolypropylene glycol, propoxypolyvinyl alcohol, or propoxypolyvinylpyrrolidone. [Item 41B] The therapeutic and / or prophylactic agent according to Item 39B or 40B, wherein the lipid moiety of the polymer-modified lipid is phosphatidylethanolamine or diacylglycerol. [Item 42B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 41B, wherein the liposome encapsulating the CHK1 inhibitor comprises: (1) 40 to 70 mol % of phospholipid, (2) 30 to 50 mol % of cholesterol, and (3) 1 to 10 mol % of a polymer-modified lipid. [Item 43B] The therapeutic and / or prophylactic agent according to any one of Items 1B to 42B, wherein the liposome further comprises an additive selected from the group consisting of inorganic acids, inorganic acid salts, organic acids, organic acid salts, sugars, buffers, antioxidants, and polymers.[Item 1C] A method for treating and / or preventing cancer in a patient exhibiting resistance to immune checkpoint inhibitor therapy, comprising administering to the patient a CHK1 inhibitor or a liposome encapsulating a CHK1 inhibitor. [Item 2C] The method according to Item 1C, characterized in that the CHK1 inhibitor and an immune checkpoint inhibitor are used in combination. [Item 3C] The method according to Item 2C, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously. [Item 4C] The method according to any one of Item 2C or 3C, characterized in that the immune checkpoint inhibitor is administered after the CHK1 inhibitor. [Item 5C] The method according to any one of Item 2C to 4C, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously, and then the anti-PD-1 antibody is administered. [Item 6C] The method according to any one of Item 2C to 4C, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously, and then the anti-PD-1 antibody is administered at least three days later. [Item 7C] The method of any one of Items 2C to 6C, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody. [Item 8C] The method of any one of Items 2C to 6C, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody. [Item 9C] The method of any one of Items 2C to 6C, wherein the anti-PD-1 antibody is pembrolizumab, nivolumab, spartalizumab, or cemiplimab. [Item 10C] The method of any one of Items 1C to 9C, wherein the number of CD3-positive cells in the tumor tissue of the cancer patient is reduced compared to tumor tissue of a patient responsive to the anti-PD-1 antibody. [Item 11C] The method of any one of Items 1C to 10C, wherein the number of CD8-positive cells in the tumor tissue of the cancer patient is reduced compared to tumor tissue of a patient responsive to the anti-PD-1 antibody. [Item 12C] The method according to any one of Items 1C to 11C, wherein the number of CD8-positive cells in the tumor tissue of the cancer patient is 0.135 or less.[Item 13C] The method of any one of Items 1C to 12C, wherein the M1-M2 ratio of macrophages in the tumor tissue of the cancer patient is 21 or less. [Item 14C] The method of any one of Items 1C to 13C, wherein the number of CD206-positive cells in the tumor tissue of the cancer patient is increased compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 15C] The method of any one of Items 1C to 14C, wherein the number of IAd-positive cells in the tumor tissue of the cancer patient is decreased compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 16C] The method of any one of Items 1C to 15C, wherein the number of CD62L-negative CD44-positive cells in the lymph nodes of the cancer patient is decreased compared to lymph nodes from patients responsive to an anti-PD-1 antibody. [Item 17C] The method of any one of Items 1C to 16C, wherein PD-L1 expression in the tumor tissue of the cancer patient is decreased compared to tumor tissue from patients responsive to an anti-PD-1 antibody. [Item 18C] The method of any one of Items 1C to 17C, wherein VEGFa expression in the tumor tissue of the cancer patient is increased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 19C] The method of any one of Items 1C to 17C, wherein Granzyme B expression in the tumor tissue of the cancer patient is decreased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 20C] The method of any one of Items 1C to 17C, wherein Interferon γ expression in the tumor tissue of the cancer patient is decreased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 21C] The method of any one of Items 1C to 17C, wherein H2abl expression in the tumor tissue of the cancer patient is decreased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 22C] The method of any one of Items 1C to 17C, wherein CD80 expression in the tumor tissue of the cancer patient is decreased compared to tumor tissue of patients responsive to an anti-PD-1 antibody.[Item 23C] The method of any one of Items 1C to 17C, characterized in that Mrc1 expression in the tumor tissue of the cancer patient is increased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 24C] The method of any one of Items 1C to 17C, characterized in that Arginase 1 expression in the tumor tissue of the cancer patient is increased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 25C] The method of any one of Items 1C to 17C, characterized in that DNA replication stress in the tumor tissue of the cancer patient is decreased compared to tumor tissue of patients responsive to an anti-PD-1 antibody. [Item 26C] A method for preventing recurrence of cancer that has gone into remission, comprising administering to a patient a CHK1 inhibitor or a liposome encapsulating a CHK1 inhibitor. [Item 27C] The method of Item 26C, characterized in that the CHK1 inhibitor is used in combination with an immune checkpoint inhibitor. [Item 28C] The method of any one of Items 26C or 27C, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody. [Item 29C] The method of any one of Items 26A or 27C, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody. [Item 30C] The method of any one of Items 26C or 27C, wherein the anti-PD-1 antibody is pembrolizumab, nivolumab, spartalizumab, or cemiplimab.[Item 31C] The cancer is selected from the group consisting of squamous cell carcinoma, melanoma, acute leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, myelodysplastic syndrome, polycythemia vera, malignant lymphoma, plasma cell neoplasm, multiple myeloma, brain tumor, head and neck cancer, head and neck squamous cell carcinoma, esophageal cancer, thyroid cancer, lung cancer, small cell lung cancer, non-small cell lung cancer, thymoma / thymic carcinoma, breast cancer, triple-negative breast cancer, gastric cancer, gallbladder / bile duct cancer, liver cancer, hepatocellular carcinoma, pancreatic cancer, colon cancer, rectal cancer, MSI-high colorectal cancer, and anal cancer. The method of any one of Items 26C to 30C, wherein the cancer is at least one type of cancer selected from the group consisting of cancer, gastrointestinal stromal tumor, chorioepithelial carcinoma, endometrial cancer, cervical cancer, ovarian cancer, platinum-resistant ovarian cancer, PARP inhibitor-resistant ovarian cancer, bladder cancer, urothelial cancer, kidney cancer, renal cell carcinoma, prostate cancer, testicular tumor, testicular germ cell tumor, ovarian germ cell tumor, Wilms' tumor, malignant melanoma, neuroblastoma, osteosarcoma, Ewing's sarcoma, chondrosarcoma, soft tissue sarcoma, and skin cancer. [Item 32C] The method of any one of Items 26C to 30C, wherein the cancer is melanoma, non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, urothelial cancer, non-muscle-invasive bladder cancer, renal cancer, hepatocellular carcinoma, MSI-high colorectal cancer, esophageal cancer, colorectal cancer, breast cancer, or endometrial cancer. [Item 33C] The method of any one of Items 26C to 30C, wherein the cancer is at least one type of cancer selected from the group consisting of head and neck cancer, squamous cell carcinoma, non-small cell lung cancer, anal cancer, breast cancer, triple-negative breast cancer, ovarian cancer, platinum-resistant ovarian cancer, and endometrial cancer. [Item 34C] The method of any one of Items 26C to 30C, wherein the cancer is a solid cancer with a high tumor mutation burden (Tumor Mutation Burden-High; TMB-High). [Item 35C] The method of any one of Items 1C to 34C, wherein the liposome further comprises a phospholipid.[Item 36C] The method according to Item 35C, wherein the phospholipid is one selected from the group consisting of phosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, soybean lecithin, egg yolk lecithin, hydrogenated egg yolk lecithin, and hydrogenated soybean lecithin, or a combination of two or more thereof. [Item 37C] The method according to any one of Items 1C to 36C, wherein the liposome further comprises a sterol. [Item 38C] The method according to any one of Items 1C to 37C, wherein the sterol is cholesterol. [Item 39C] The method according to any one of Items 1C to 36C, wherein the liposome further comprises a polymer-modified lipid. [Item 40C] The method according to Item 39C, wherein the polymer moiety of the polymer-modified lipid is polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, methoxypolyethylene glycol, methoxypolypropylene glycol, methoxypolyvinyl alcohol, methoxypolyvinylpyrrolidone, ethoxypolyethylene glycol, ethoxypolypropylene glycol, ethoxypolyvinyl alcohol, ethoxypolyvinylpyrrolidone, propoxypolyethylene glycol, propoxypolypropylene glycol, propoxypolyvinyl alcohol, or propoxypolyvinylpyrrolidone. [Item 41C] The method according to Item 39C or 40C, wherein the lipid moiety of the polymer-modified lipid is phosphatidylethanolamine or diacylglycerol. [Item 42C] The method according to any one of Items 1C to 41C, wherein the liposome encapsulating the CHK1 inhibitor comprises: (1) 40 to 70 mol % of a phospholipid, (2) 30 to 50 mol % of cholesterol, and (3) 1 to 10 mol % of a polymer-modified lipid. [Item 43C] The method according to any one of Items 1C to 42C, wherein the liposome further comprises an additive selected from the group consisting of inorganic acids, inorganic acid salts, organic acids, organic acid salts, sugars, buffers, antioxidants, and polymers. [Item 44] The CHK1 inhibitor is a compound represented by formula (1): [In the formula, R １ represents a hydrogen atom, an optionally substituted C１－６ Alkyl, optionally substituted C ３－１０ Cycloalkyl, optionally substituted 3- to 10-membered saturated heterocyclic group, optionally substituted C ６－１０ R represents an aryl or an optionally substituted 5- to 12-membered heteroaryl; ２ represents a hydrogen atom, a halogen atom, a cyano, a nitro, a carboxyl, a sulfonic acid, a phosphoric acid, or -OR ３ , -SR ３ , -COR ４ , -CO ２ R ４ , -CONR ５ R ６ , -SO ２ R ４ , -SO ２ NR ５ R ６ , -OCOR ４ , -OCO ２ R ４ , -OCONR ５ R ６ , -NR ５ R ６ , -NR ７ COR ４ , -NR ７ CO ２ R ４ , -NR ７ CONR ５ R ６ , -NR ７ SO ２ R ４ , -NR ７ SO ２ NR ５ R ６ , optionally substituted C １－６ Alkyl, optionally substituted C ２－６ Alkenyl, optionally substituted C ２－６ Alkynyl, optionally substituted C ３－１０ Cycloalkyl, optionally substituted 3- to 10-membered saturated heterocyclic group, optionally substituted C ６－１０ R represents an aryl or an optionally substituted 5- to 12-membered heteroaryl; ３ is a hydrogen atom or C １－６ represents alkyl, R ４ is C１－６ represents alkyl, R ５ , R ６ and R ７ are each independently a hydrogen atom or C １－６ alkyl, wherein R ５ and R ６ Both are C １－６ When X, Y and Z are alkyl, they may be taken together with the nitrogen atom to which they are bonded to form a 3- to 8-membered nitrogen-containing saturated heterocycle, and X, Y and Z are each independently CR ８ or a nitrogen atom, wherein X, Y, and Z simultaneously represent CR ８ Not R ８ When there are a plurality of groups, each group independently represents a hydrogen atom, a halogen atom, a cyano group, a nitro group, a carboxyl group, a sulfonic acid group, a phosphoric acid group, or an —OR ９ , -SR ９ , -COR １０ , -CO ２ R １０ , -CONR １１ R １２ , -SO ２ R １０ , -SO ２ NR １１ R １２ , -OCOR １０ , -OCO ２ R １０ , -OCONR １１ R １２ , -NR １１ R １２ , -NR １３ COR １０ , -NR １３ CO ２ R １０ , -NR １３ CONR １１ R １２ , -NR １３ SO ２ R １０ , -NR １３ SO ２ NR １１ R １２ , optionally substituted C １－６ Alkyl, optionally substituted C ２－６ Alkenyl, optionally substituted C２－６ Alkynyl, optionally substituted C ３－１０ Cycloalkyl, optionally substituted 3- to 10-membered saturated heterocyclic group, optionally substituted C ６－１０ R represents an aryl or an optionally substituted 5- to 12-membered heteroaryl; ９ is a hydrogen atom or C １－６ represents alkyl, R １０ is C １－６ represents alkyl, R １１ , R １２ and R １３ are each independently a hydrogen atom or C １－６ alkyl, wherein R １１ and R １２ Both are C １－６ When L is alkyl, they may be taken together with the nitrogen atom to which they are bonded to form a 3- to 8-membered nitrogen-containing saturated heterocycle, and L is a single bond or an optionally substituted C １－６ represents alkylene; V represents a single bond, optionally substituted C ３－１０ represents a cycloalkylene or an optionally substituted 3- to 10-membered divalent saturated heterocyclic group; W represents a single bond or an optionally substituted C １－６ represents an alkylene; Q represents a hydrogen atom or NHR １４ represents R １４ represents a hydrogen atom, an optionally substituted C １－６ Alkyl, optionally substituted C ３－１０ [Item 45] The therapeutic and / or prophylactic agent according to any one of Items 1 to 43, which is a compound represented by the formula:

[0023] or a pharmaceutically acceptable salt thereof, wherein R １ , R ２ , R ８ , R １４ Optionally substituted C in L, V, and W １－６ Alkyl, optionally substituted C ２－６ Alkenyl, optionally substituted C ２－６ Alkynyl, optionally substituted C ３－１０Cycloalkyl, optionally substituted 3- to 10-membered saturated heterocyclic group, optionally substituted C ６－１０ Aryl, optionally substituted 5- to 12-membered heteroaryl, optionally substituted C １－６ Alkylene, optionally substituted C ３－１０ The cycloalkylene or optionally substituted 3- to 10-membered divalent saturated heterocyclic group is each independently (1) a halogen atom, (2) a hydroxyl group, or (3) C ６－１０ (4) 5- to 12-membered heteroaryl; (5) C １－６ alkyl, (6) C ２－６ alkenyl, (7) C ２－６ alkynyl, (8) C １－６ Alkoxy, (9) C １－６ Alkylthio (10) C ３－１０ (11) a 3- to 10-membered saturated heterocyclic group, (12) a carboxyl, (13) -COR １５ , (14)-CO ２ R １５ , (15)-CONR １６ R １７ , (16)-NR １６ R １７ , (17)-NR １８ COR １５ , (18)-NR １８ CO ２ R １５ , (19)-NR １８ SO ２ R １５ , (20)-NR １８ CONR １６ R １７ , (21)-NR １８ SO ２ NR １６ R １７ , (22)-SO ２ R １５ , (23)-SO ２ NR １６ R １７ , (24)-OCOR １５ , (25)-OCO ２ R１５ , (26)-OCONR １６ R １７ (27) sulfonic acid, (28) phosphoric acid, (29) cyano, and (30) nitro, wherein the (3) C ６－１０ (4) 5- to 12-membered heteroaryl; (5) C １－６ alkyl, (6) C ２－６ alkenyl, (7) C ２－６ alkynyl, (8) C １－６ Alkoxy, (9) C １－６ Alkylthio, (9) C ３－１０ The group shown in cycloalkyl and (10) 3- to 10-membered saturated heterocyclic group is (a) a halogen atom, (b) a hydroxyl group, or (c) C ６－１０ (d) 5- to 12-membered heteroaryl; (e) C １－６ (f) alkyl, ２－６ alkenyl, (g) C ２－６ alkynyl, (h) C １－６ Alkoxy, (i) C ３－１０ (j) a 3- to 10-membered saturated heterocyclic group; (k) a carboxyl; (l) —COR １５ , (m)-CO ２ R １５ , (n)-CONR １６ R １７ , (o)-NR １６ R １７ , (p)-NR １８ COR １５ , (q)-NR １８ SO ２ R １５ , (r)-SO ２ R １５ , (s)-SO ２ NR １６ R １７ (t) sulfonic acid, (u) phosphate, (v) cyano, and (w) nitro; R １５ However, if there are multiple, each independently, C１－６ alkyl, R １６ and R １７ are each independently a hydrogen atom or C １－６ alkyl, and R １６ or R １７ If there are multiple １６ or R １７ may be the same or different, and R １６ and R １７ Both are C １－６ When R is alkyl, they may be combined with the nitrogen atom to which they are bonded to form a 3- to 8-membered nitrogen-containing saturated heterocycle, １８ is a hydrogen atom or C １－６ Item 46. The therapeutic and / or prophylactic agent according to Item 44, wherein R is alkyl. １ is a C optionally substituted with a hydrogen atom or 1 to 3 fluorine atoms １－６ The therapeutic and / or prophylactic agent according to any one of Items 44 to 45, wherein R is alkyl. [Item 47] １ [Item 48] The therapeutic and / or prophylactic agent according to any one of Items 44 to 46, wherein R is a methyl group. ２ is a hydrogen atom, a halogen atom, cyano, -OR ３ , C １－６ Alkyl, C ３－１０ The therapeutic and / or prophylactic agent according to any one of Items 44 to 47, wherein R is a cycloalkyl or a 3- to 10-membered saturated heterocyclic group. [Item 49] ２ The therapeutic and / or prophylactic agent according to any one of Items 44 to 46, wherein R is cyano. [Item 50] ８ When there are a plurality of groups, each group independently represents a hydrogen atom, a halogen atom, a cyano group, or an -OR ９ , -CO ２ R １０ , -CONR １１ R １２ , -NR １１ R １２ , -NR １３ COR １０ , C １－６Alkyl (the alkyl is a fluorine atom, a chlorine atom, a bromine atom, a hydroxyl group, a phenyl, a 5- to 6-membered heteroaryl, C １－６ Alkoxy, C ３－７ cycloalkyl, 3- to 7-membered saturated heterocyclic group, —CONR １６ R １７ , -NR １６ R １７ and cyano), C ３－１０ Cycloalkyl (the cycloalkyl is a fluorine atom, a chlorine atom, a bromine atom, a hydroxyl group, a phenyl, a 5- to 6-membered heteroaryl, C １－６ Alkyl, C １－６ Alkoxy, C ３－７ cycloalkyl, 3- to 7-membered saturated heterocyclic group, —CONR １６ R １７ , -NR １６ R １７ and cyano), a 3- to 10-membered saturated heterocyclic group (the saturated heterocyclic group is optionally substituted with 1 to 3 identical or different substituents selected from the group consisting of a fluorine atom, a chlorine atom, a bromine atom, a hydroxyl group, phenyl, a 5- or 6-membered heteroaryl, C １－６ Alkyl, C １－６ Alkoxy, C ３－７ cycloalkyl, 3- to 7-membered saturated heterocyclic group, —CONR １６ R １７ , -NR １６ R １７ and cyano), phenyl (the phenyl is optionally substituted with 1 to 3 identical or different substituents selected from the group consisting of a fluorine atom, a chlorine atom, a bromine atom, a hydroxyl group, phenyl, 5- to 6-membered heteroaryl, C １－６ Alkyl, C １－６ Alkoxy, C ３－７ cycloalkyl, 3- to 7-membered saturated heterocyclic group, —CONR １６ R １７ , -NR １６ R １７and cyano), or 5- to 6-membered heteroaryl (the heteroaryl is a fluorine atom, a chlorine atom, a bromine atom, a hydroxyl group, phenyl, a 5- to 6-membered heteroaryl, C １－６ Alkyl, C １－６ Alkoxy, C ３－７ cycloalkyl, 3- to 7-membered saturated heterocyclic group, —CONR １６ R １７ , -NR １６ R １７ [Item 51] The therapeutic and / or prophylactic agent according to any one of Items 44 to 49, wherein L is a single bond, or C １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ [Item 52] The therapeutic and / or prophylactic agent according to any one of Items 44 to 50, wherein V is a single bond, C ３－１０ Cycloalkylene (the cycloalkylene is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), or a 3- to 10-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a fluorine atom, a hydroxyl group, a C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ [Item 53] The therapeutic and / or prophylactic agent according to any one of Items 44 to 51, wherein W is a single bond, or C １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R１７ [Item 54] The therapeutic and / or prophylactic agent according to any one of Items 44 to 52, wherein R is a substituted or unsubstituted alkyl group, and R is a substituted or unsubstituted alkyl group, and R is a substituted or unsubstituted alkyl group. １４ is a hydrogen atom, C １－６ Alkyl (the alkyl is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), C ３－１０ Cycloalkyl (the cycloalkyl is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), or a 3- to 10-membered saturated heterocyclic group (the saturated heterocyclic group may be substituted with 1 to 3 identical or different substituents selected from the group consisting of a fluorine atom, a hydroxyl group, a C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ [Item 55] The therapeutic and / or prophylactic agent according to any one of Items 44 to 53, wherein formula (1) is the following formula (2): wherein X, Y, and Z each independently represent CR ８ or a nitrogen atom, wherein X, Y, and Z simultaneously represent CR ８ Not R ８ When there are a plurality of atoms, each independently represents a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C. １－６ Alkyl (the alkyl is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), or a 5- or 6-membered heteroaryl (the heteroaryl may be substituted with 1 or 2 identical or different substituents selected from the group consisting of a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR１６ R １７ and cyano), wherein L represents a single bond, or １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), V represents a single bond, C ３－１０ Cycloalkylene (the cycloalkylene is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), or a 3- to 10-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a fluorine atom, a hydroxyl group, a C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), W represents a single bond, or C １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), Q is a hydrogen atom or NHR １４ represents R １４ is a hydrogen atom, C １－６ Alkyl (the alkyl is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), C ３－１０ Cycloalkyl (the cycloalkyl is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３Alkoxy, —NR １６ R １７ and cyano), or a 3- to 10-membered saturated heterocyclic group (the saturated heterocyclic group may be substituted with 1 to 3 identical or different substituents selected from the group consisting of a fluorine atom, a hydroxyl group, a C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), R １６ and R １７ are each independently a hydrogen atom or C １－６ represents alkyl, and R １６ or R １７ If there are multiple １６ or R １７ may be the same or different, and R １６ and R １７ Both are C １－６ [Item 56] The therapeutic and / or prophylactic agent according to any one of Items 44 to 55, wherein one or two of X, Y and Z represent a nitrogen atom. [Item 57] X is a nitrogen atom, and Y and Z are CR ８ [Item 58] The therapeutic and / or prophylactic agent according to any one of Items 44 to 56, wherein Y is a nitrogen atom, and X and Z are CR ８ [Item 59] The therapeutic and / or prophylactic agent according to any one of Items 44 to 56, wherein Z is a nitrogen atom, and X and Y are CR ８ [Item 60] The therapeutic and / or prophylactic agent according to any one of Items 44 to 56, wherein R ８ When there are a plurality of groups, each group independently represents a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－６ Alkyl (the alkyl is a fluorine atom, a hydroxyl group, and １－３[Item 61] The therapeutic and / or prophylactic agent according to any one of Items 44 to 59, wherein L is a single bond, or C １－６ The therapeutic and / or prophylactic agent according to any one of Items 44 to 60, wherein V is alkylene (the alkylene may be substituted with 1 to 2 identical or different substituents selected from the group consisting of a fluorine atom, a hydroxyl group, and a cyano). [Item 62] V is a single bond, C ３－７ Cycloalkylene (the cycloalkylene is a C group optionally substituted with a fluorine atom, a hydroxyl group, or 1 to 2 hydroxyl groups) １－３ a 3- to 7-membered divalent saturated heterocyclic group (which may be substituted with 1 to 2 identical or different substituents selected from the group consisting of alkyl, and cyano), or a 3- to 7-membered divalent saturated heterocyclic group (which may be substituted with a fluorine atom, a hydroxyl group, 1 to 2 hydroxyl groups or fluorine atoms, １－３ [Item 63] The therapeutic and / or prophylactic agent according to any one of Items 44 to 61, wherein W is a single bond, or C １－６ The therapeutic and / or prophylactic agent according to any one of Items 44 to 62, wherein R is alkylene (the alkylene may be substituted with 1 to 2 identical or different substituents selected from the group consisting of a fluorine atom, a hydroxyl group, and a cyano). [Item 64] R １４ is a hydrogen atom, or C １－６ The therapeutic and / or prophylactic agent according to any one of Items 44 to 63, wherein Q is alkyl (which may be substituted with 1 to 2 identical or different substituents selected from the group consisting of a fluorine atom, a hydroxyl group, and a cyano). [Item 65] Q is a hydrogen atom, NH ２ [Item 66] The therapeutic and / or prophylactic agent according to any one of Items 44 to 64, wherein formula (1) is the following formula (3): [In the formula, R ８ａ and R ８ｂ are each independently a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－３L represents a single bond or a C alkyl group optionally substituted with one hydroxyl group. １－６ represents an alkylene; V represents a single bond; ３－７ Cycloalkylene (the cycloalkylene is a hydroxyl group and a C １－３ alkyl), or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is substituted with a fluorine atom, a cyano, a hydroxyl group, and a C １－３ and W represents a single bond or a C alkyl group optionally substituted with one hydroxyl group. １－３ represents alkylene, Q is a hydrogen atom or NH ２ [Item 67] The therapeutic and / or prophylactic agent according to Item 44, wherein R ８ａ and R ８ｂ [Item 68] The therapeutic and / or prophylactic agent according to Item 66, wherein R is each independently a hydrogen atom, a fluorine atom, or a chlorine atom. ８ａ and R ８ｂ [Item 69] The therapeutic and / or prophylactic agent according to Item 66, wherein each of L is independently a hydrogen atom or a chlorine atom. １－３ [Item 70] The therapeutic and / or prophylactic agent according to any one of Items 66 to 68, wherein V is alkylene. [Item 71] The therapeutic and / or prophylactic agent according to any one of Items 66 to 69, wherein V is a single bond. [Item 72] The therapeutic and / or prophylactic agent according to any one of Items 66 to 69, wherein V is C ３－７ [Item 72] The therapeutic and / or prophylactic agent according to any one of Items 66 to 71, wherein W is cycloalkylene. [Item 73] The therapeutic and / or prophylactic agent according to any one of Items 66 to 71, wherein W is a single bond. [Item 74] The therapeutic and / or prophylactic agent according to any one of Items 66 to 71, wherein W is C １－３ [Item 74] The therapeutic and / or prophylactic agent according to any one of Items 66 to 71, wherein Q is alkylene. ２ [Item 75] The therapeutic and / or prophylactic agent according to any one of Items 66 to 73, wherein formula (1) is the following formula (4): [In the formula, R ８ｂ and R ８ｃare each independently a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－３ L represents a single bond or C １－６ alkylene (which may be substituted with one substituent selected from the group consisting of a fluorine atom, a hydroxyl group, and a cyano group); V is a single bond; C ３－７ Cycloalkylene (the cycloalkylene is a hydroxyl group and a C １－３ alkyl), or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a C group optionally substituted with a fluorine atom, a cyano, a hydroxyl group, and 1 to 3 hydroxyl groups and fluorine atoms). １－３ and W represents a single bond or a C alkyl group optionally substituted with one hydroxyl group. １－３ represents alkylene, Q is a hydrogen atom, NH ２ Item 76] R ８ｂ and R ８ｃ [Item 77] The therapeutic and / or prophylactic agent according to Item 75, wherein L is a single bond, or C optionally substituted with one hydroxyl group or fluorine atom. １－６ Item 78: The therapeutic and / or prophylactic agent according to any one of Items 75 and 76, wherein V is alkylene. [Item 79] The therapeutic and / or prophylactic agent according to any one of Items 76 and 77, wherein V is a single bond, C ３－７ Cycloalkylene, or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a fluorine atom, a hydroxyl group, and a C １－３ [Item 79] The therapeutic and / or prophylactic agent according to any one of Items 75 to 77, wherein W is a single bond, or C １－３ [Item 80] The therapeutic and / or prophylactic agent according to any one of Items 75 to 78, wherein Q is alkylene. [Item 81] The therapeutic and / or prophylactic agent according to any one of Items 75 to 79, wherein Q is a hydrogen atom. [Item 82] The therapeutic and / or prophylactic agent according to any one of Items 75 to 79, wherein Q is NH ２[Item 82] The therapeutic and / or prophylactic agent according to any one of Items 75 to 79, wherein Q is NHMe. [Item 83] The therapeutic and / or prophylactic agent according to any one of Items 75 to 79, wherein formula (1) is the following formula (5): [In the formula, R ８ａ and R ８ｃ are each independently a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－３ L represents a single bond or a C alkyl group optionally substituted with one hydroxyl group. １－6 represents an alkylene; V represents a single bond; ３－７ Cycloalkylene (the cycloalkylene is a hydroxyl group and a C １－３ alkyl), or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a C １－３ and W represents a single bond or a C alkyl group optionally substituted with one hydroxyl group. １－３ represents alkylene, Q is a hydrogen atom or NH ２ [Item 84] The therapeutic and / or prophylactic agent according to Item 44, wherein R ８ａ and R ８ｃ [Item 85] The therapeutic and / or prophylactic agent according to Item 83, wherein L is a C optionally substituted with one hydroxyl group. １－6 Item 86] The therapeutic and / or prophylactic agent according to any one of Items 83 and 84, wherein V is alkylene. [Item 87] The therapeutic and / or prophylactic agent according to any one of Items 83 and 84, wherein V is a single bond, C ３－７ Cycloalkylene or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a C group optionally substituted with a hydroxyl group and one hydroxyl group). １－３ [Item 87] The therapeutic and / or prophylactic agent according to any one of Items 83 to 85, wherein W is a single bond, or C １－３[Item 88] The therapeutic and / or prophylactic agent according to any one of Items 83 to 86, wherein Q is alkylene. [Item 89] The therapeutic and / or prophylactic agent according to any one of Items 83 to 87, wherein Q is a hydrogen atom. [Item 90] The therapeutic and / or prophylactic agent according to any one of Items 83 to 87, wherein Q is NH ２ [Item 90] The therapeutic and / or prophylactic agent according to any one of Items 83 to 87, wherein formula (1) is the following formula (6): [In the formula, R ８ａ is a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－３ L represents a single bond or C １－６ alkylene (which may be substituted with one substituent selected from the group consisting of a fluorine atom, a hydroxyl group, and a cyano group); V is a single bond; C ３－７ Cycloalkylene (the cycloalkylene is a hydroxyl group and a C １－３ alkyl), or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a C group optionally substituted with a fluorine atom, a cyano, a hydroxyl group, and 1 to 3 fluorine atoms). １－３ and W represents a single bond or a C alkyl group optionally substituted with one hydroxyl group. １－３ represents alkylene, Q is a hydrogen atom, NH ２ Item 91] R ８ａ [Item 92] The therapeutic and / or prophylactic agent according to Item 90, wherein L is a hydrogen atom. １－４ Item 93: The therapeutic and / or prophylactic agent according to any one of items 90 and 91, wherein V is alkylene. [Item 94] V is a single bond, or C ３－７ The therapeutic and / or prophylactic agent according to any one of Items 90 to 92, wherein W is a single bond, or C １－３ The therapeutic and / or prophylactic agent according to any one of Items 90 to 93, wherein Q is alkylene. [Item 95] The therapeutic and / or prophylactic agent according to any one of Items 90 to 93, wherein Q is NH ２Item 95. The therapeutic and / or prophylactic agent according to any one of Items 90 to 94, [Item 96] The therapeutic and / or prophylactic agent according to Item 44, which contains a compound selected from the following compounds or a pharmaceutically acceptable salt thereof: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[2-(3-aminopropoxy)-6-chloro-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[3-(3-aminopropoxy)-5-methoxypyridin-4-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[4-(3-aminopropoxy)-2-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, 5-[(5-{3-[(3-fluoroazetidin-3-yl)methoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{2-methoxy-4-[(3-methylazetidin-3-yl)methoxy]pyridin-3-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{4-[(3-hydroxyazetidin-3-yl)methoxy]-2-methoxypyridin-3-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-{[5-(4-{[3-(hydroxymethyl)azetidin-3-yl]methoxy}-2-methoxypyridin-3-yl)-1H-pyrazol-3-yl]amino}pyrazine-2-carbonitrile, 5-[(5-{3-[(3R)-3-aminobutoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(3S)-3-aminobutoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-{[5-(3-{[1-(aminomethyl)cyclopropyl]methoxy}-5-methoxypyridin-4-yl)-1H-pyrazol-3-yl]amino}pyrazine-2-carbonitrile,5-[(5-{3-methoxy-5-[(morpholin-2-yl)methoxy]pyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-methoxy-5-[(morpholin-2-yl)methoxy]pyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(2S)-3-amino-2-hydroxypropoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, N-{5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}-5-chloropyrazin-2-amine, N-{5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}-5-(trifluoromethyl)pyrazin-2-amine, 5-({5-[4-(3-aminopropoxy)-6-methoxypyrimidin-5-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[3-(azetidin-3-yl)methoxy-5-methoxypyridin-4-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, 5-{[5-(3-{[(1R,3S)-3-aminocyclohexyl]oxy}-5-methoxypyridin-4-yl)-1H-pyrazol-3-yl]amino}pyrazine-2-carbonitrile, (S)-5-[(5-{3-[(3-fluoropyrrolidin-3-yl)methoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, (S)-5-[(5-{3-methoxy-[5-(pyrrolidin-3-yl)methoxy]pyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, (R)-5-[(5-{3-[(3-fluoropyrrolidin-3-yl)methoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-{[5-(4-{[1-(aminomethyl)cyclopropyl]methoxy}-6-methoxypyrimidin-yl)-1H-pyrazol-3-yl]amino}pyrazine-2-carbonitrile,5-({5-[3-(3-aminopropoxy)-5-(fluoromethoxy)pyridin-4-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, 5-[(5-{3-methoxy-5-[(3-methylazetidin-3-yl)methoxy]pyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-{[5-(3-{[3-(difluoromethyl)azetidin-3-yl]methoxy}-5-methoxypyridin-4-yl)-1H-pyrazol-3-yl]amino}pyrazine-2-carbonitrile, 5-[(5-{3-[(2S)-3-amino-2-methylpropoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(2R)-3-amino-2-methylpropoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(2S)-3-amino-2-fluoropropoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(2R)-3-amino-2-fluoropropoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(2R)-3-amino-2-fluoropropoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-methoxy-5-[3-(methylamino)propoxy]pyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-{[5-(3-{[(1R,3R)-3-aminocyclopentyl]oxy}-5-methoxypyridin-4-yl)-1H-pyrazol-3-yl]amino}pyrazine-2-carbonitrile, 5-[(5-{3-[(1R)-1-(azetidin-3-yl)ethoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(1R)-1-(3-hydroxyazetidin-3-yl)ethoxy]-5-methoxypyridin-4-yl}-1H-pyrazol-3-yl)amino]pyrazine-2-carbonitrile,5-{[5-(3-methoxy-5-{[(1R,3R)-3-(methylamino)cyclopentyl]oxy}pyridin-4-yl)-1H-pyrazol-3-yl]amino}pyrazine-2-carbonitrile, 5-{[5-(3-{[(1R,2S,4S,5S)-4-aminobicyclo[3.1.0]hexan-2-yl]oxy}-5-methoxypyridin-4-yl)-1H-pyrazol-3-yl]amino}pyrazine-2-carbonitrile, 5-{[5-(2-{[1-(aminomethyl)cyclopropyl]methoxy}-4-methoxypyridin-3-yl)-1H-pyrazol-3-yl]amino}pyrazine-2-carbonitrile, 5-{[5-(4-{[1-(aminomethyl)cyclopropyl]methoxy}-2-methoxypyridin-3-yl)-1H-pyrazol-3-yl]amino}pyrazine-2-carbonitrile. [Item 97] A therapeutic and / or prophylactic agent according to Item 44, comprising a compound selected from the following compounds or a pharmaceutically acceptable salt thereof: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[3-(3-aminopropoxy)-5-methoxypyridin-4-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[4-(3-aminopropoxy)-2-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile. [Item 98] The therapeutic and / or prophylactic agent according to Item 44, which contains a compound selected from the following compounds or a pharmaceutically acceptable salt thereof: 5-({5-[4-(3-aminopropoxy)-2-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile. [Item 99] The therapeutic and / or prophylactic agent according to Item 44, which contains the following compound or a pharmaceutically acceptable salt thereof: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile. [Item 100] The therapeutic and / or prophylactic agent according to Item 44, which contains the following compound or a pharmaceutically acceptable salt thereof:5-({5-[3-(3-aminopropoxy)-5-methoxypyridin-4-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile.

[0010] The present disclosure provides a therapeutic and / or prophylactic agent for cancer patients exhibiting resistance to immune checkpoint inhibitors, which contains a CHK1 inhibitor or a liposome encapsulating a CHK1 inhibitor as an active ingredient. Furthermore, when the inhibitor or liposome is used in combination with an immune checkpoint inhibitor, a superior antitumor effect is exhibited compared to when the inhibitor or liposome is used alone. Furthermore, the inhibitor, the liposome, or a combination of these with an immune checkpoint inhibitor also exhibits an effect of preventing the recurrence of cancer in remission.

[0011] The present disclosure provides a CHK1 inhibitor comprising a 5-heteroaryl-1H-pyrazol-3-amine derivative or a pharmaceutically acceptable salt thereof. The compound of the present disclosure combines excellent CHK1 inhibitory activity with high safety. When encapsulated in liposomes and released sustainedly from the liposomes, the compound acts on cancer sustainably and exhibits potent antitumor effects. The compound of the present disclosure is useful as a therapeutic agent for diseases involving CHK1, and specifically, is applicable to patients with pancreatic cancer, ovarian cancer, osteosarcoma, Ewing's sarcoma, chondrosarcoma, soft tissue sarcoma, and the like.

[0012] The present disclosure has demonstrated pharmacological activity in Phase 2 studies in ovarian cancer patients. Compared to prexasertib, a 5-phenyl-1H-pyrazol-3-amine derivative, the present disclosure failed to maximize efficacy with 72 hours of continuous exposure, as predicted from non-clinical studies. It was revealed, however, that the compound or salt thereof of the present disclosure can further improve this. Furthermore, while prexasertib causes side effects due to high maximum blood concentrations after a single administration, the compound or salt thereof of the present disclosure did not exhibit such side effects. While side effects due to prolonged drug exposure were observed after three consecutive days of administration, the compound or salt thereof of the present disclosure did not exhibit such side effects (Non-Patent Documents 3 and 6). Additionally, the present inventors have newly discovered that prexasertib poses a risk of hepatotoxicity, and the compound of the present invention exhibits a reduced risk of hepatotoxicity. From the above, the compound of the present disclosure possesses both excellent CHK1 inhibitory activity and a high level of safety.

[0013] In particular, compounds having a pyridine ring with a nitrogen atom at a specific position, as represented by formula (3), are characterized by having both high antitumor activity and safety, as well as excellent pharmacokinetics.

[0014] In particular, the compound of Example 1, which is included in formula (3), exhibits higher CHK1 inhibitory activity and has a strong cell proliferation inhibitory effect compared to prexasertib, and is characterized by being highly safe. Furthermore, the compound of Example 1 exhibits good pharmacokinetics and is characterized by being efficiently encapsulated in liposomes. A liposome formulation of the compound of Example 1 has excellent antitumor effect and is highly safe, and when used in combination with an existing anticancer drug, it exhibits even more exceptional antitumor effect.

[0015]

[0039] Figure 7 shows the powder X-ray diffraction pattern of Form I of the compound of Example 39. The horizontal axis represents the diffraction angle 2θ (°), and the vertical axis represents the number of counts (the same applies to Figures 2 to 6 below). Figure 8 shows the powder X-ray diffraction pattern of Form II of the compound of Example 40. Figure 9 shows the powder X-ray diffraction pattern of Form III of the compound of Example 41. Figure 10 shows the powder X-ray diffraction pattern of Form IV of the compound of Example 42. Figure 11 shows the powder X-ray diffraction pattern of Form V of the compound of Example 43. Figure 12 shows the powder X-ray diffraction pattern of Form VI of the compound of Example 44. Figure 7 shows the tumor PD response effect of a solution formulation or liposome formulation of plexasertib in ES-2 tumor-bearing mice (Test A of Test Example 11). The PD response was evaluated by detecting the expression levels of γH2AX and tubulin by Western blotting, quantifying the intensity of each band using ImageJ software, and calculating the relative value of the expression level of γH2AX based on the expression level of tubulin. The vertical axis indicates the relative value of γH2AX. Figure 8 shows the tumor PD response effect using ES-2 cancer-bearing mice for Test B of Test Example 11, a solution formulation of plexasertib, or the liposome formulation of Example 1. The PD response was evaluated by detecting the expression levels of γH2AX and tubulin by Western blotting, quantifying the intensity of each band using ImageJ software, and calculating the relative value of the expression level of γH2AX based on the expression level of tubulin. The vertical axis indicates the relative value of γH2AX. Figure 9 shows the tumor PD response effect using ES-2 cancer-bearing mice for Test C of Test Example 11, a liposome formulation of Example 1. The PD response was evaluated by detecting the expression levels of γH2AX and tubulin by Western blotting, quantifying the intensity of each band using ImageJ software, and calculating the relative value of the expression level of γH2AX based on the expression level of tubulin. The vertical axis shows the relative value of γH2AX. Figure 10 shows an example of an administration regimen for the anti-PD-1 antibody and the liposome preparation of Example 1. The liposome preparation of Example 1 is administered simultaneously with the anti-PD-1 antibody, and then administered once a week (A1), once every two weeks (A2), once every three weeks (A3), once every four weeks (A4), or once every five weeks (A5).The anti-PD-1 antibody is administered simultaneously with Example 1, and then administered every 2 weeks (B1), 3 weeks (B2), 4 weeks (B3), or 6 weeks (B4) according to the individual regimens. That is, Example 1 and the anti-PD-1 antibody are administered according to an administration regimen selected by any combination thereof. Preferred examples include combinations of administration regimens of A1 and B1, A1 and B2, A1 and B3, A1 and B4, A1 and B5, A2 and B1, A2 and B2, A2 and B3, A2 and B4, A2 and B5, A3 and B1, A3 and B2, A3 and B3, A3 and B4, A3 and B5, A4 and B1, A4 and B2, A4 and B3, A4 and B4, A4 and B5, A5 and B1, A5 and B2, A5 and B3, A5 and B4, and A5 and B5. Fig. 11 is a diagram showing the cell counts of CD8-positive T cells per clonal amount and the clonotypes of the TCR repertoire for the liposome preparation of Example 1 in the test of Test Example 28. Fig. 12 is a diagram showing the CD8 score for the liposome preparation of Example 1 in the test of Test Example 28. Fig. 13 is a diagram showing the M1-M2 ratio for the liposome preparation of Example 1 in the test of Test Example 28. Fig. 14 is a diagram showing the expression level of ssDNA by the treatment of Example 1 in the test of Test Example 34.

[0016] The present disclosure will be described in more detail below. Throughout this specification, unless otherwise specified, singular expressions should be understood to include the plural concept. Therefore, singular articles (e.g., "a," "an," "the," etc. in English) should be understood to include the plural concept unless otherwise specified. Furthermore, it should be understood that terms used in this specification are used in the sense commonly used in the relevant field unless otherwise specified. Therefore, unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In the case of conflict, the present specification (including definitions) will prevail.

[0017] The terms used in this specification are explained below.

[0018] In this specification, the number of substituents in a group defined as "optionally substituted" is not particularly limited as long as substitution is possible. Furthermore, unless otherwise specified, the description of each group also applies to the case where the group is a part or substituent of another group.

[0019] Examples of the "halogen atom" include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom. A fluorine atom or a chlorine atom is preferred.

[0020] "C １－６ "Alkyl" means an alkyl having 1 to 6 carbon atoms, ６ "Alkyl" means alkyl having 6 carbon atoms, and similarly for other numbers.

[0021] "C １－６ "Alkyl" means a straight or branched chain saturated hydrocarbon group having from 1 to 6 carbon atoms. １－６ As the alkyl, preferably "C １－４ alkyl", and more preferably "C １－３ "C alkyl" is an example. １－３ Specific examples of "alkyl" include methyl, ethyl, propyl, 1-methylethyl, etc. １－４ Specific examples of "alkyl" include the above-mentioned "C １－３ In addition to the specific examples of "alkyl", butyl, 1,1-dimethylethyl, 1-methylpropyl, 2-methylpropyl, etc. are also included. １－６ Specific examples of "alkyl" include the above-mentioned "C １－４ In addition to the specific examples of "alkyl", pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 1-methylbutyl, 2-methylbutyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, hexyl, and the like can be mentioned.

[0022] "C ２－６ "Alkenyl" means a straight or branched chain unsaturated hydrocarbon group having 2 to 6 carbon atoms and containing 1 to 3 double bonds. ２－６ Alkenyl is preferably "C ２－４"C alkenyl" is an example. ２－４ Specific examples of "alkenyl" include vinyl, propenyl, methylpropenyl, butenyl, etc. ２－６ Specific examples of "alkenyl" include the above-mentioned "C ２－４ In addition to the specific examples of "alkenyl", pentenyl, hexenyl, etc. are also included.

[0023] "C ２－６ "Alkynyl" means a straight or branched chain unsaturated hydrocarbon group having 2 to 6 carbon atoms and containing one triple bond. ２－６ As the "alkynyl", preferably "C ２－４ "C alkynyl" is an example. ２－４ Specific examples of "alkynyl" include propynyl, methylpropynyl, butynyl, etc. ２－６ Specific examples of "alkynyl" include the above-mentioned "C ２－４ In addition to the specific examples of "alkynyl", methylbutynyl, pentynyl, hexynyl, etc. are included.

[0024] "C １－６ "Alkoxy" means "C １－６ "C alkyloxy" １－６ The "C alkyl" moiety is １－６ "C" is synonymous with "alkyl." １－６ The "alkoxy" is preferably "C １－４ Alkoxy" is preferred, and "C １－３ "Alkoxy" is an example. １－３ Specific examples of "alkoxy" include methoxy, ethoxy, propoxy, 1-methylethoxy, etc. １－４ Specific examples of "alkoxy" include the above-mentioned "C １－３ In addition to the specific examples of "alkoxy", butoxy, 1,1-dimethylethoxy, 1-methylpropoxy, 2-methylpropoxy, etc. are also included. １－６ Specific examples of "alkoxy" include the above-mentioned "C １－４In addition to the specific examples of "alkoxy", pentyloxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, 1-methylbutoxy, 2-methylbutoxy, 4-methylpentyloxy, 3-methylpentyloxy, 2-methylpentyloxy, 1-methylpentyloxy, hexyloxy, and the like can be mentioned.

[0025] "C １－６ "C" in "Alkylthio" １－６ The "C alkyl" moiety is １－６ "C" is synonymous with "alkyl." １－６ As the alkylthio, preferably "C １－４ alkylthio", and more preferably "C １－３ "Alkylthio" is an example. １－３ Specific examples of "alkylthio" include methylthio, ethylthio, propylthio, 1-methylethylthio, etc. １－４ Specific examples of "alkylthio" include the above-mentioned "C １－３ In addition to the specific examples of "alkylthio," butylthio, 1,1-dimethylethylthio, 1-methylpropylthio, 2-methylpropylthio, etc. are also included. １－６ Specific examples of "alkylthio" include the above-mentioned "C １－４ In addition to the specific examples of "alkylthio", pentylthio, 1,1-dimethylpropylthio, 1,2-dimethylpropylthio, 1-methylbutylthio, 2-methylbutylthio, 4-methylpentylthio, 3-methylpentylthio, 2-methylpentylthio, 1-methylpentylthio, hexylthio, and the like can be mentioned.

[0026] "C １－６ "Alkylene" means a divalent saturated hydrocarbon group having 1 to 6 carbon atoms, which may be straight or branched. １－６ As the "alkylene", preferably "C １－４ alkylene", and more preferably "C １－３ "C alkylene" is an example. １－３ Specific examples of "alkylene" include methylene, ethylene, propylene, trimethylene, etc. １－４Specific examples of "alkylene" include the above-mentioned "C １－３ In addition to the specific examples of "alkylene," butylene, 1,1-dimethylethylene, 1,2-dimethylethylene, 1-methyltrimethylene, 2-methyltrimethylene, etc. are also included. １－６ Specific examples of "alkylene" include the above-mentioned "C １－４ In addition to the specific examples of "alkylene," examples include pentylene, 1,1-dimethyltrimethylene, 1,2-dimethyltrimethylene, 1-methylbutylene, 2-methylbutylene, 1-methylpentylene, 2-methylpentylene, 3-methylpentylene, hexylene, and the like.

[0027] "C ３－１０ "Cycloalkyl" means a cyclic saturated hydrocarbon group having 3 to 10 carbon atoms, and includes those having a partially unsaturated bond and those having a bridged structure. ３－１０ As the "cycloalkyl", preferably "C ３－７ "Cycloalkyl" is an example. ３－７ Specific examples of "cycloalkyl" include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc. ３－１０ Specific examples of "cycloalkyl" include the above-mentioned "C ３－７ In addition to the specific examples of "cycloalkyl", cyclooctyl, cyclononyl, cyclodecyl, adamantyl, etc. are included.

[0028] Also, "C ３－１０ "Cycloalkyl" includes the above C ３－１０ Bicyclic compounds in which a cycloalkyl and an aromatic hydrocarbon ring are fused together are also included. Specific examples of such fused ring compounds include the structures shown below.

[0029] Specific examples of the crosslinked structure include the structures shown below.

[0030] "C ３－１０"Cycloalkylene" means a cyclic divalent saturated hydrocarbon group having 3 to 10 carbon atoms, and includes those having a partially unsaturated bond and those having a bridged structure. ３－１０ As the "cycloalkylene", preferably "C ３－７ "Cycloalkylene" is an example. ３－７ Specific examples of "cycloalkylene" include cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene, etc. ３－１０ Specific examples of "cycloalkyl" include the above-mentioned "C ３－７ In addition to the specific examples of "cycloalkyl", cyclooctylene, cyclononylene, cyclodecylene, adamantylene, etc. are included.

[0031] Specific examples of the crosslinked structure include the structures shown below.

[0032] The term "3- to 10-membered saturated carbocyclic ring" refers to a cyclic saturated hydrocarbon having 3 to 10 carbon atoms. A preferred example of the "3- to 10-membered saturated carbocyclic ring" is a "4- to 6-membered saturated carbocyclic ring." Specific examples of the "4- to 6-membered saturated carbocyclic ring" include a cyclobutane ring, a cyclopentane ring, and a cyclohexane ring. Specific examples of the "3- to 10-membered saturated carbocyclic ring" include, in addition to the specific examples of the "4- to 6-membered saturated carbocyclic ring" mentioned above, a cyclopropane ring, a cycloheptane ring, cyclooctane, cyclononane, and cyclodecane.

[0033] The term "3- to 10-membered saturated heterocyclic group" refers to a monovalent saturated heterocyclic group consisting of 1 to 2 atoms independently selected from the group consisting of nitrogen, oxygen, and sulfur atoms, and 2 to 9 carbon atoms, and includes groups having a partially unsaturated bond and a bridged structure. The atoms constituting the ring are -C(O)-, -S(O)-, -SO ２-, and may include those oxidized to -. As the "3- to 10-membered saturated heterocyclic group", preferably, a "4- to 7-membered monocyclic saturated heterocyclic group" is mentioned. Specific examples of the "4- to 7-membered monocyclic saturated heterocyclic group" include, for example, oxetanyl, azetidinyl, tetrahydrofuryl, pyrrolidinyl, imidazolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, dioxothiomorpholinyl, hexamethyleneiminyl, oxazolidinyl, thiazolidinyl, oxoimidazolidinyl, dioxoimidazolidinyl, oxooxazolidinyl, dioxooxazolidinyl, dioxothiazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, and the like. Examples of the "3- to 10-membered saturated heterocyclic group" include oxiranyl, aziridinyl, etc., in addition to the specific examples of the "4- to 7-membered monocyclic saturated heterocyclic group". The "3- to 10-membered saturated heterocyclic group" also includes bicyclic groups in which the 3- to 10-membered saturated heterocyclic group and a 6-membered aromatic hydrocarbon ring or a 6-membered aromatic heterocyclic ring form a fused ring. Examples of the 6-membered aromatic hydrocarbon ring forming a fused ring include a benzene ring. Examples of the 6-membered aromatic heterocyclic ring forming a fused ring include pyridine, pyrimidine, pyridazine, etc. Specific examples of the bicyclic "3- to 10-membered saturated heterocyclic group" forming a fused ring include dihydroindolyl, dihydroisoindolyl, dihydropurinyl, dihydrothiazolopyrimidinyl, dihydrobenzodioxanyl, isoindolyl, indazolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydronaphthyridinyl, and the like.

[0034] The term "3- to 8-membered nitrogen-containing saturated heterocycle" refers to a saturated heterocycle consisting of one nitrogen atom and 2 to 7 carbon atoms. A preferred example of the "3- to 8-membered nitrogen-containing saturated heterocycle" is a "4- to 6-membered nitrogen-containing saturated heterocycle". Specific examples of the "4- to 6-membered nitrogen-containing saturated heterocycle" include an azetidine ring, a pyrrolidine ring, and a piperidine ring. Specific examples of the "3- to 8-membered nitrogen-containing saturated heterocycle" include, in addition to the specific examples of the "4- to 6-membered nitrogen-containing saturated heterocycle", an aziridine ring, an azepane ring, and an azocane ring.

[0035] The term "3- to 10-membered divalent saturated heterocyclic group" refers to a divalent saturated heterocyclic group consisting of 1 to 2 atoms independently selected from the group consisting of nitrogen, oxygen, and sulfur atoms, and 2 to 9 carbon atoms, and includes those having a partially unsaturated bond and those having a bridged structure. The atoms constituting the ring are -C(O)-, -S(O)-, -SO ２ -, and may include those oxidized such as -. As the "3- to 10-membered saturated heterocyclic group", preferably, a "4- to 7-membered monocyclic saturated heterocyclic group" is mentioned. Specific examples of the "4- to 7-membered monocyclic saturated heterocyclic group" include, for example, oxetanylene, azetidinylene, tetrahydrofurylene, pyrrolidinylene, imidazolidinylene, piperidinylene, morpholinylene, thiomorpholinylene, dioxothiomorpholinylene, hexamethyleneiminylene, oxazolidinylene, thiazolidinylene, oxoimidazolidinylene, dioxoimidazolidinylene, oxooxazolidinylene, dioxooxazolidinylene, dioxothiazolidinylene, tetrahydrofuranylene, tetrahydropyranylene, and the like. Examples of the "3- to 10-membered saturated heterocyclic group" include oxiranylene, aziridinylene, etc., in addition to the specific examples of the "4- to 7-membered monocyclic saturated heterocyclic group". The "3- to 10-membered saturated heterocyclic group" also includes bicyclic groups in which the 3- to 10-membered saturated heterocyclic group and a 6-membered aromatic hydrocarbon ring or a 6-membered aromatic heterocyclic ring form a fused ring. Examples of the 6-membered aromatic hydrocarbon ring forming a fused ring include a benzene ring. Examples of the 6-membered aromatic heterocyclic ring forming a fused ring include pyridine, pyrimidine, pyridazine, etc. Specific examples of the bicyclic "3- to 10-membered saturated heterocyclic group" forming a fused ring include dihydroindolylene, dihydroisoindolylene, dihydropurinylene, dihydrothiazolopyrimidinylene, dihydrobenzodioxanylene, isoindolylene, indazolylene, tetrahydroquinolinylene, tetrahydroisoquinolinylene, tetrahydronaphthyridinylene, and the like.

[0036] "C ６－１０ "Aryl" means an aromatic hydrocarbon ring group having 6 to 10 carbon atoms. ６－１０Specific examples of "aryl" include phenyl, 1-naphthyl, 2-naphthyl, etc. Preferably, phenyl is used. ６－１０ "Aryl" includes the above C ６－１０ Aryl and C ４－６ Also included are bicyclic rings formed by condensing with a cycloalkyl or a 5- to 6-membered saturated heterocycle. ６－１０ Specific examples of "aryl" include groups shown below.

[0037] The "aromatic hydrocarbon ring" is defined as the ring ６－１０ "aryl" means the ring portion of an "aryl";

[0038] "5- to 12-membered heteroaryl" means a monocyclic 5- to 7-membered aromatic heterocyclic ring group or a bicyclic 8- to 12-membered aromatic heterocyclic ring group containing 1 to 4 atoms independently selected from the group consisting of nitrogen, oxygen, and sulfur atoms. A "5- to 7-membered monocyclic heteroaryl" is preferred. More preferred is pyridyl, pyrimidinyl, quinolyl, or isoquinolyl. Even more preferred is pyridyl. Specific examples of a "5- to 7-membered monocyclic heteroaryl" include pyridyl, pyridazinyl, isothiazolyl, pyrrolyl, furyl, thienyl, thiazolyl, imidazolyl, pyrimidinyl, thiadiazolyl, pyrazolyl, oxazolyl, isoxazolyl, pyrazinyl, triazinyl, triazolyl, oxadiazolyl, triazolyl, and tetrazolyl. Specific examples of the "5- to 12-membered heteroaryl" include, in addition to the specific examples of the "5- to 7-membered monocyclic heteroaryl", indolyl, indazolyl, chromenyl, quinolyl, isoquinolyl, benzofuranyl, benzothienyl, benzoxazolyl, benzothiazolyl, benzisoxazolyl, benzisothiazolyl, benzotriazolyl, benzimidazolyl, etc.

[0039] The term "aromatic heterocycle" refers to the ring moiety of the above "5- to 12-membered heteroaryl".

[0040] "Cancer" and "tumor" are used interchangeably in this disclosure and both refer to malignant tumors, including carcinomas, sarcomas, and hematological malignancies. Specific examples of "cancer" or "tumor" include, for example, acute leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, myelodysplastic syndrome, polycythemia vera, malignant lymphoma, plasma cell neoplasm, multiple myeloma, brain tumor, head and neck cancer, esophageal cancer, thyroid cancer, small cell lung cancer, non-small cell lung cancer, thymoma / thymic carcinoma, breast cancer, gastric cancer, gallbladder / bile duct cancer, liver cancer, hepatocellular carcinoma, pancreatic cancer, colon cancer, rectal cancer, anal cancer, gastrointestinal stromal tumor, chorioepithelial carcinoma, uterine cancer, cervical cancer, ovarian cancer, bladder cancer, urothelial cancer, kidney cancer, renal cell carcinoma, prostate cancer, testicular tumor, testicular germ cell tumor, ovarian germ cell tumor, Wilms' tumor, malignant melanoma, neuroblastoma, osteosarcoma, Ewing's sarcoma, chondrosarcoma, soft tissue sarcoma, and skin cancer.

[0041] Prexasertib is a compound having the following structure:

[0042] In the compounds of the present disclosure represented by formula (1), (2), (3), (4), or (5), R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R ８ａ , R ８ｂ , R ８ｃ , R ９ , R １０ , R １１ , R １２ , R １３ , R １４ , R １５ , R １６ , R １７ , R １８ Preferred values ​​for X, Y, Z, L, V, W, Q and Y are as follows, but the technical scope of the present disclosure is not limited to the range of compounds listed below.

[0043] R １ A preferred embodiment of the formula (I) is a C group optionally substituted with 1 to 3 fluorine atoms. １－６ alkyl.１ More preferred embodiments of R include a methyl group or an ethyl group which may be substituted with 1 to 3 fluorine atoms. １ A more preferred embodiment of R is a methyl group optionally substituted with 1 to 3 fluorine atoms. １ An even more preferred embodiment of is a methyl group.

[0044] R ２ Preferred embodiments of the group include a hydrogen atom, a halogen atom, cyano, -OR ３ , C optionally substituted with 1 to 3 halogen atoms １－６ Alkyl, C ３－１０ cycloalkyl, or a 3- to 10-membered saturated heterocyclic group. ２ A more preferred embodiment of the above is a C group optionally substituted with a hydrogen atom, a halogen atom, cyano, or 1 to 3 halogen atoms. １－６ alkyl. ２ A more preferred embodiment of the formula (I) is a C group optionally substituted with a halogen atom, cyano, or 1 to 3 halogen atoms. １－６ alkyl. ２ An even more preferred embodiment of is cyano.

[0045] R ３ A preferred embodiment of is a hydrogen atom or C １－６ alkyl. ３ A more preferred embodiment of the above is C １－６ alkyl. ３ A more preferred embodiment of the formula is C １－３ alkyl. ３ An even more preferred embodiment of is a methyl group.

[0046] R ４ A preferred embodiment of the formula is C １－３ alkyl. ４ A more preferred embodiment of is a methyl group.

[0047] R ５ , R ６ and R ７A preferred embodiment of is a hydrogen atom or C １－６ alkyl. ５ , R ６ and R ７ A more preferred embodiment of the above is C １－６ alkyl. ５ , R ６ and R ７ A more preferred embodiment of the formula is C １－３ alkyl. ５ , R ６ and R ７ An even more preferred embodiment of is a methyl group.

[0048] R ８ Preferred embodiments of the group include a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, and C １－６ Alkyl (the alkyl is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), or 5- or 6-membered heteroaryl (the heteroaryl is optionally substituted with 1 to 2 identical or different substituents selected from the group consisting of a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano). ８ More preferred embodiments of the group include a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, and C １－６ Alkyl (the alkyl is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano). ８ In a more preferred embodiment, the group is a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－３ alkyl. ８ An even more preferred embodiment of is a hydrogen atom or a chlorine atom.

[0049] R ８ａ , R ８ｂ and R ８ｃ Preferred embodiments of R include a hydrogen atom, a fluorine atom, a chlorine atom, and a bromine atom. ８ａ , R ８ｂ and R ８ｃ More preferred embodiments of R include a hydrogen atom and a chlorine atom. ８ａ , R ８ｂ and R ８ｃ A more preferred embodiment of is a hydrogen atom.

[0050] R ９ A preferred embodiment of is a hydrogen atom or C １－６ alkyl. ９ A more preferred embodiment of the above is C １－３ alkyl. ９ A more preferred embodiment of is a methyl group.

[0051] R １０ A preferred embodiment of the formula is C １－３ alkyl. １０ A more preferred embodiment of is a methyl group.

[0052] R １１ , R １２ and R １３ A preferred embodiment of is a hydrogen atom or C １－６ alkyl. １１ , R １２ and R １３ A more preferred embodiment of the above is C １－３ alkyl. １１ , R １２ and R １３ A more preferred embodiment of is a methyl group.

[0053] R １４ Preferred embodiments of the group include a hydrogen atom, C １－６ Alkyl (the alkyl is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７and cyano), C ３－１０ Cycloalkyl (the cycloalkylene is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), or a 3- to 10-membered saturated heterocyclic group (the saturated heterocyclic group may be substituted with 1 to 3 identical or different substituents selected from the group consisting of a fluorine atom, a hydroxyl group, a C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano). １４ More preferred embodiments of the group include a hydrogen atom, C １－６ Alkyl (the alkyl is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano). １４ In a more preferred embodiment, it is a hydrogen atom or C １－３ alkyl (the alkyl may be substituted with one substituent selected from the group consisting of a fluorine atom, a hydroxyl group, and a cyano). １４ An even more preferred embodiment of is a hydrogen atom or a methyl group.

[0054] R １５ A preferred embodiment of the formula is C １－６ alkyl. １５ A more preferred embodiment of the above is C １－３ alkyl. １５ A more preferred embodiment of is a methyl group.

[0055] R １６ and R １７ A preferred embodiment of is a hydrogen atom or C １－６ alkyl. １６ and R１７ A more preferred embodiment of the above is C １－６ alkyl. １６ and R １７ A more preferred embodiment of the formula is C １－３ alkyl. １６ and R １７ An even more preferred embodiment of is a methyl group.

[0056] R １８ A preferred embodiment of is a hydrogen atom or C １－６ alkyl. １８ A more preferred embodiment of the above is C １－６ alkyl. １８ A more preferred embodiment of the formula is C １－３ alkyl. １８ An even more preferred embodiment of is a methyl group.

[0057] Preferred embodiments of X, Y and Z include CR ８ In the present disclosure, when referring to a compound or a pharmaceutically acceptable salt of the present disclosure, X, Y, and Z may simultaneously be CR ８ Another preferred embodiment of X, Y, and Z is that one or two of X, Y, and Z represent a nitrogen atom. Another preferred embodiment of X, Y, and Z is that X is CR ８ and Y and Z are nitrogen atoms. In another preferred embodiment of X, Y and Z, Y is CR ８ and X and Z are nitrogen atoms. In another preferred embodiment of X, Y, and Z, Z is CR ８ and X and Y are nitrogen atoms.

[0058] Preferred embodiments of L include a single bond or C １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７and cyano). A more preferred embodiment of L is a single bond or C １－６ An alkylene group (the alkylene group may be substituted with one substituent selected from the group consisting of a fluorine atom, a hydroxyl group, and a cyano group) is preferred. A more preferred embodiment of L is a single bond or C １－６ An even more preferred embodiment of L is a single bond or a C alkylene group optionally substituted with one hydroxyl group. １－６ Examples include alkylene.

[0059] Preferred embodiments of V include a single bond, C ３－１０ Cycloalkylene (the cycloalkylene is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), or a 3- to 10-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a fluorine atom, a hydroxyl group, a C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano). ３－７ Cycloalkylene (the cycloalkylene is a C group optionally substituted with a fluorine atom, a hydroxyl group, or 1 to 2 hydroxyl groups) １－３ and cyano), or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a fluorine atom, a hydroxyl group, a C １－３(V may be substituted with 1 to 2 identical or different substituents selected from the group consisting of alkyl, and cyano). ３－７ Cycloalkylene (the cycloalkylene is a C group optionally substituted with a fluorine atom, a hydroxyl group, or 1 to 2 hydroxyl groups or fluorine atoms) １－３ alkyl), or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a C １－３ and alkyl, each of which may be substituted with 1 to 2 identical or different substituents selected from the group consisting of alkyl. ３－７ Cycloalkylene (the cycloalkylene is a hydroxyl group and a C １－３ alkyl), or a 3- to 7-membered divalent saturated heterocyclic group (the 3- to 7-membered saturated heterocyclic group is substituted with a fluorine atom, a cyano, a hydroxyl group, and a C １－３ and alkyl.

[0060] Preferred embodiments of W include a single bond or C １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano). １－３ An alkylene group (the alkylene group may be substituted with one substituent selected from the group consisting of a fluorine atom, a hydroxyl group, and a cyano group) is preferred. １－３ An alkylene group (the alkylene group may be substituted with one substituent selected from the group consisting of a fluorine atom and a hydroxyl group) is preferred. An even more preferred embodiment of W is a single bond or a C group optionally substituted with one hydroxyl group. １－３ Examples include alkylene.

[0061] Preferred embodiments of Q include a hydrogen atom or NHR １４ More preferred embodiments of Q include a hydrogen atom, NH ２ In a more preferred embodiment, Q is a hydrogen atom or NH ２ An even more preferred embodiment of Q is a hydrogen atom. Another even more preferred embodiment of Q is NH ２ Examples include:

[0062] One embodiment of the compound represented by formula (1) is the following (A): (A) R １ is a methyl group or an ethyl group optionally substituted with 1 to 3 fluorine atoms, and R ２ is a hydrogen atom, a halogen atom, cyano, -OR ３ , C optionally substituted with 1 to 3 halogen atoms １－６ Alkyl, C ３－１０ cycloalkyl or a 3- to 10-membered saturated heterocyclic group, R ３ is a hydrogen atom or C １－６ alkyl; X, Y, and Z are each independently CR ８ or a nitrogen atom, wherein X, Y, and Z simultaneously represent CR ８ Not R ８ is a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, C １－６ Alkyl (the alkyl is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), or 5- or 6-membered heteroaryl (the heteroaryl is optionally substituted with 1 to 2 identical or different substituents selected from the group consisting of a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), and L is a single bond or C １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C１－３ Alkoxy, —NR １６ R １７ and cyano), and V is a single bond, C ３－１０ Cycloalkylene (the cycloalkylene is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), or a 3- to 10-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a fluorine atom, a hydroxyl group, a C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), and W is a single bond or C １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), and Q is a hydrogen atom or NHR １４ and R １４ is a hydrogen atom, C １－６ Alkyl (the alkyl is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), C ３－１０ Cycloalkyl (the cycloalkylene is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), or a 3- to 10-membered saturated heterocyclic group (the saturated heterocyclic group may be substituted with 1 to 3 identical or different substituents selected from the group consisting of a fluorine atom, a hydroxyl group, a C １－３ Alkyl, C１－３ Alkoxy, —NR １６ R １７ and cyano), and R １６ and R １７ is a hydrogen atom or C １－６ or a pharmaceutically acceptable salt thereof.

[0063] One embodiment of the compound represented by formula (1) is the following (B): (B) R １ is a methyl group optionally substituted with 1 to 3 fluorine atoms, and R ２ is a C optionally substituted with a halogen atom, cyano, or 1 to 3 halogen atoms; １－６ alkyl; X, Y, and Z are each independently CR ８ or a nitrogen atom, wherein X, Y, and Z simultaneously represent CR ８ Not R ８ is a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－３ alkyl, and L is a single bond or C １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), and V is a single bond, C ３－１０ Cycloalkylene (the cycloalkylene is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), or a 3- to 10-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a fluorine atom, a hydroxyl group, a C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７and cyano), and W is a single bond or C １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), and Q is a hydrogen atom or NHR １４ and R １４ is a hydrogen atom, C １－６ Alkyl (the alkyl is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), C ３－１０ Cycloalkyl (the cycloalkylene is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), or a 3- to 10-membered saturated heterocyclic group (the saturated heterocyclic group may be substituted with 1 to 3 identical or different substituents selected from the group consisting of a fluorine atom, a hydroxyl group, a C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), and R １６ and R １７ is a hydrogen atom or C １－６ or a pharmaceutically acceptable salt thereof.

[0064] One embodiment of the compound represented by formula (1) is the following (C): (C) R １ is a methyl group or a fluoromethyl group, and R ２ is a chlorine atom, cyano, or trifluoromethyl group; X, Y, and Z are each independently CR ８ or a nitrogen atom, wherein X, Y, and Z simultaneously represent CR ８Not R ８ is a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－３ alkyl, and L is a single bond or C １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), and V is a single bond, C ３－１０ Cycloalkylene (the cycloalkylene is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), or a 3- to 10-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a fluorine atom, a hydroxyl group, a C １－３ Alkyl, C １－３ Alkoxy, —NR １６ R １７ and cyano), and W is a single bond or C １－６ Alkylene (the alkylene is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, —NR １６ R １７ and cyano), and Q is a hydrogen atom or NHR １４ and R １４ is a hydrogen atom or C １－３ alkyl (the alkyl may be substituted with one substituent selected from the group consisting of a fluorine atom, a hydroxyl group, and a cyano); R １６ and R １７ is a hydrogen atom or C １－６ or a pharmaceutically acceptable salt thereof, wherein:

[0065] One embodiment of the compound represented by formula (1) is the following (D): (D) R１ is a methyl group, and R ２ is cyano; X, Y, and Z are each independently CR ８ or a nitrogen atom, wherein X, Y, and Z simultaneously represent CR ８ Not R ８ is a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－３ alkyl, and L is a single bond or C １－６ alkylene (the alkylene may be substituted with one substituent selected from the group consisting of a fluorine atom, a hydroxyl group, and a cyano), and V is a single bond, C ３－７ Cycloalkylene (the cycloalkylene is a C group optionally substituted with a fluorine atom, a hydroxyl group, or 1 to 2 hydroxyl groups) １－３ and cyano), or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a fluorine atom, a hydroxyl group, a C １－３ and C is a single bond, or C １－３ alkylene (the alkylene may be substituted with one substituent selected from the group consisting of a fluorine atom, a hydroxyl group, and a cyano), and Q is a hydrogen atom, NH ２ or NHMe, or a pharmaceutically acceptable salt thereof.

[0066] One embodiment of the compound represented by formula (1) or (2) is the following (E): (E) X, Y, and Z are each independently CR ８ or a nitrogen atom, wherein X, Y, and Z simultaneously represent CR ８ Not R ８ is a hydrogen atom or a chlorine atom, and L is a single bond or a C optionally substituted with one hydroxyl group or one fluorine atom. １－６ alkylene, V is a single bond, C ３－７ Cycloalkylene (the cycloalkylene is a hydroxyl group and a C １－３alkyl), or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a C group optionally substituted with a fluorine atom, cyano, a hydroxyl group, and 1 to 2 hydroxyl groups or fluorine atoms). １－３ and W is a single bond or a C alkyl group optionally substituted with one hydroxyl group. １－３ alkylene, Q is a hydrogen atom, NH ２ or NHMe, or a pharmaceutically acceptable salt thereof.

[0067] One embodiment of the compound represented by formula (3) is the following (F): (F) R ８ａ and R ８ｂ are each independently a hydrogen atom or a chlorine atom, L is a single bond or C optionally substituted with one hydroxyl group １－３ alkylene, V is a single bond, C ３－７ Cycloalkylene (the cycloalkylene is a hydroxyl group and a C １－３ alkyl), or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is substituted with a fluorine atom, a cyano, a hydroxyl group, and a C １－３ and W is a C alkyl group optionally substituted with a single bond or one hydroxyl group. １－３ alkylene, Q is a hydrogen atom or NH ２ or a pharmaceutically acceptable salt thereof.

[0068] One embodiment of the compound represented by formula (3) is (G) below. (G) R ８ａ and R ８ｂ are each independently a hydrogen atom or a chlorine atom, and L is C optionally substituted with one hydroxyl group. １－３ alkylene, V is a single bond, W is a single bond or a C group optionally substituted with one hydroxyl group. １－３ alkylene, and Q is NH ２or a pharmaceutically acceptable salt thereof.

[0069] One embodiment of the compound represented by formula (3) is (H) below. (H) R ８ａ and R ８ｂ are each independently a hydrogen atom, and L is C optionally substituted with one hydroxyl group. １－３ alkylene, and V is C ３－７ cycloalkylene, W is a C optionally substituted with one hydroxyl group; １－３ alkylene, and Q is NH ２ or a pharmaceutically acceptable salt thereof.

[0070] One embodiment of the compound represented by formula (4) is the following (I): (I) R ８ｂ and R ８ｃ is a hydrogen atom, L is a single bond, or C optionally substituted with one hydroxyl group or one fluorine atom; １－３ alkylene, V is a single bond, C ３－７ Cycloalkylene (the cycloalkylene is a hydroxyl group and a C １－３ alkyl), or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a C group optionally substituted with a fluorine atom, cyano, a hydroxyl group, and 1 to 3 hydroxyl groups or fluorine atoms). １－３ and W is a C alkyl group optionally substituted with a single bond or one hydroxyl group. １－３ alkylene, Q is a hydrogen atom, NH ２ or NHMe, or a pharmaceutically acceptable salt thereof.

[0071] One embodiment of the compound represented by formula (4) is the following (J): (J) R ８ｂ and R ８ｃ are each independently a hydrogen atom, and L is C optionally substituted with one hydroxyl group or fluorine atom. １－３alkylene, and V is a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a C group optionally substituted with a fluorine atom, a cyano, a hydroxyl group, or 1 to 3 hydroxyl groups and fluorine atoms). １－３ and R is an integer of 1 to 10, and R is an integer of 1 to 10. In one embodiment, R is an integer of 1 to 10, and R is an integer of 1 to 10.

[0072] One embodiment of the compound represented by formula (4) is (K) below. (K) R ８ｂ and R ８ｃ are each independently a hydrogen atom, and L is C optionally substituted with one hydroxyl group or fluorine atom. １－３ alkylene, and V is C ３－７ Cycloalkylene (the cycloalkylene is a hydroxyl group and a C １－３ and W is C optionally substituted with one hydroxyl group. １－３ alkylene, and Q is NH ２ or a pharmaceutically acceptable salt thereof.

[0073] One embodiment of the compound represented by formula (4) is (L) below. (L) R ８ｂ and R ８ｃ are each independently a hydrogen atom, L is a single bond, and V is C ３－７ Cycloalkylene (the cycloalkylene is a hydroxyl group and a C １－３ alkyl), W is a single bond, and Q is NH ２ or NHMe, or a pharmaceutically acceptable salt thereof.

[0074] One embodiment of the compound represented by formula (4) is (M) below. (M) R ８ｂ and R ８ｃ are each independently a hydrogen atom, and L is C optionally substituted with one hydroxyl group or fluorine atom. １－３alkylene, V is a single bond, W is a single bond or a C group optionally substituted with one hydroxyl group. １－３ alkylene, and Q is NH ２ or NHMe, or a pharmaceutically acceptable salt thereof.

[0075] One embodiment of the compound represented by formula (5) is (N) below. (N) R ８ａ and R ８ｃ are each independently a hydrogen atom, L is a single bond or C optionally substituted with one hydroxyl group １－３ alkylene, V is a single bond, C ３－７ Cycloalkylene (the cycloalkylene is a hydroxyl group and a C １－３ alkyl), or a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a C １－３ and W is a single bond or a C alkyl group optionally substituted with one hydroxyl group. １－３ alkylene, Q is a hydrogen atom or NH ２ or a pharmaceutically acceptable salt thereof.

[0076] One embodiment of the compound represented by formula (5) is the following (O): (O) R ８ａ and R ８ｃ are each independently a hydrogen atom, and L is C optionally substituted with one hydroxyl group. １－３ alkylene, and V is a 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group is a C １－３ and R is an integer of 1 to 10, and R is an integer of 1 to 10. In one embodiment, R is an integer of 1 to 10, and R is an integer of 1 to 10.

[0077] One embodiment of the compound represented by formula (5) is the following (P): (P) R ８ａand R ８ｃ are each independently a hydrogen atom, and L is C optionally substituted with one hydroxyl group. １－３ alkylene, V is a single bond, W is a single bond or a C group optionally substituted with one hydroxyl group. １－３ alkylene, and Q is NH ２ or a pharmaceutically acceptable salt thereof.

[0078] One embodiment of the compound represented by formula (5) is (Q) below. (Q) R ８ａ are each independently a hydrogen atom, and L is C optionally substituted with one hydroxyl group. １－３ alkylene, and V is C ３－７ cycloalkylene, W is a C optionally substituted with one hydroxyl group; １－３ alkylene, and Q is NH ２ or a pharmaceutically acceptable salt thereof.

[0079] One embodiment of the compound represented by formula (6) is the following (R): (R) R ８ａ is a hydrogen atom, and L is C １－３ alkylene, and V is a single bond or C ３－７ cycloalkylene, W is a single bond or C １－３ alkylene, and Q is NH ２ or a pharmaceutically acceptable salt thereof.

[0080] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile hydrochloride (Form I) having an X-ray powder diffraction pattern with characteristic peaks at least at 7.2°±0.2° in terms of 2θ.

[0081] A further aspect of the present invention includes 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile hydrochloride in a crystalline form (Form I) having an X-ray powder diffraction pattern with characteristic peaks at least at 8.8°±0.2° in terms of 2θ.

[0082] A further aspect of the present invention includes 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile hydrochloride in a crystalline form (Form I) having an X-ray powder diffraction pattern with characteristic peaks at least at 7.2°±0.2° and 8.8°±0.2° 2θ.

[0083] A further aspect of the present invention includes crystalline Form I of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile hydrochloride, having a powder X-ray diffraction pattern with characteristic peaks at 7.2°±0.2°, 8.8°±0.2°, 9.8°±0.2°, 10.2°±0.2°, 10.7°±0.2°, 16.7°±0.2°, 18.5°±0.2°, 26.2°±0.2°, 27.0°±0.2°, and 26.4°±0.2°, expressed in terms of 2θ. The crystal is identified by the presence of four or five peaks selected from these ten peaks.

[0084] A further aspect of the present invention includes 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile phosphate in a crystalline form (Form II) having an X-ray powder diffraction pattern with characteristic peaks at least at 6.8°±0.2° in terms of 2θ.

[0085] A further aspect of the present invention includes 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile phosphate in a crystalline form (Form II) having an X-ray powder diffraction pattern with characteristic peaks at least at 13.0°±0.2° in 2θ.

[0086] A further aspect of the present invention includes 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile phosphate in a crystalline form (Form II) having an X-ray powder diffraction pattern with characteristic peaks at least at 6.8°±0.2° and 13.0°±0.2° 2θ.

[0087] A further aspect of the present invention includes crystalline Form II of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile phosphate, which has a powder X-ray diffraction pattern with characteristic peaks at the following angles in degrees 2θ: 6.8°±0.2°, 7.5°±0.2°, 11.7°±0.2°, 11.9°±0.2°, 13.0°±0.2°, 16.4°±0.2°, 19.3°±0.2°, 20.4°±0.2°, 22.7°±0.2°, and 24.3°±0.2°. The crystal is identified by the presence of four or five peaks selected from these ten peaks.

[0088] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile tosylate (Form III) having an X-ray powder diffraction pattern with characteristic peaks at least at 6.0°±0.2° in 2θ.

[0089] A further aspect of the present invention includes a crystalline form (Form III) of the tosylate salt of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile having an X-ray powder diffraction pattern with characteristic peaks at least at 17.0°±0.2° in terms of 2θ.

[0090] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile tosylate (Form III) having an X-ray powder diffraction pattern with characteristic peaks at least at 6.0°±0.2° and 17.0°±0.2° 2-theta.

[0091] A further aspect of the present invention includes 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile tosylate in the crystalline form of Form III, which has a powder X-ray diffraction pattern with characteristic peaks at 6.0°±0.2°, 9.0°±0.2°, 12.1°±0.2°, 14.4°±0.2°, 16.2°±0.2°, 17.0°±0.2°, 22.8°±0.2°, and 26.3°±0.2° in terms of 2θ. The crystal is identified by the presence of four or five peaks selected from these ten peaks.

[0092] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile (Form IV) having an X-ray powder diffraction pattern with characteristic peaks at least at 9.3°±0.2° in terms of 2θ.

[0093] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile (Form IV) having an X-ray powder diffraction pattern with characteristic peaks at least at 10.2°±0.2° in terms of 2θ.

[0094] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile (Form IV) having an X-ray powder diffraction pattern with characteristic peaks at least at 9.3°±0.2° and 10.2°±0.2° 2-theta.

[0095] A further aspect of the present invention includes crystalline Form IV of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, having a powder X-ray diffraction pattern with characteristic peaks at 9.3°±0.2°, 10.2°±0.2°, 10.7°±0.2°, 13.6°±0.2°, 16.7°±0.2°, 17.1°±0.2°, 17.8°±0.2°, 18.6°±0.2°, 26.1°±0.2°, and 26.4°±0.2°, expressed in terms of 2θ. The crystal is identified by the presence of four or five peaks selected from these ten peaks.

[0096] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile (Form V) having an X-ray powder diffraction pattern with characteristic peaks at least at 7.9°±0.2° expressed in terms of 2θ.

[0097] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile (Form V) having an X-ray powder diffraction pattern with characteristic peaks at least at 8.7°±0.2° in terms of 2θ.

[0098] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile (Form V) having an X-ray powder diffraction pattern with characteristic peaks at least at 7.9°±0.2° and 8.7°±0.2° 2θ.

[0099] A further aspect of the present invention includes crystalline Form V of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, having a powder X-ray diffraction pattern with characteristic peaks at the following angles in degrees 2-theta: 7.9°±0.2°, 8.7°±0.2°, 12.2°±0.2°, 13.1°±0.2°, 15.9°±0.2°, 17.6°±0.2°, 19.9°±0.2°, 21.9°±0.2°, 22.8°±0.2°, and 26.6°±0.2°. The crystal is identified by the presence of four or five peaks selected from these ten peaks.

[0100] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile (Form VI) having an X-ray powder diffraction pattern with characteristic peaks at least at 5.3°±0.2° in terms of 2θ.

[0101] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile (Form VI) having an X-ray powder diffraction pattern with characteristic peaks at least at 5.7°±0.2° expressed in terms of 2θ.

[0102] A further aspect of the present invention includes a crystalline form of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile (Form VI) having an X-ray powder diffraction pattern with characteristic peaks at least at 5.3°±0.2° and 5.7°±0.2° 2-theta.

[0103] A further aspect of the present invention includes crystalline Form VI of 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile, having an X-ray powder diffraction pattern with characteristic peaks at 5.3°±0.2°, 5.7°±0.2°, 7.0°±0.2°, 7.3°±0.2°, 7.8°±0.2°, 8.4°±0.2°, 9.3°±0.2°, 10.5°±0.2°, 11.5°±0.2°, and 14.1°±0.2° in terms of 2-theta. The crystal is identified by the presence of four or five peaks selected from these ten peaks.

[0104] When administering the compound of the present disclosure, the dosage will vary depending on the symptoms, age, administration method, etc., but for example, in the case of intravenous injection, an effect can be expected by administering a daily dose of 0.01 mg (preferably 0.1 mg) as a lower limit to 1000 mg (preferably 100 mg) as an upper limit to an adult in a single dose or in divided doses depending on the symptoms. Examples of administration schedules include a single dose, once daily administration for three consecutive days, or twice daily administration for one week. Furthermore, each of the administration methods described above can be repeated at intervals of about 1 day to about 60 days.

[0105] The compounds of the present disclosure may be administered parenterally or orally, but are preferably administered parenterally, more preferably by intravenous injection. The compounds of the present disclosure are also preferably formulated and administered in a pharmaceutically acceptable carrier, such as liposomes.

[0106] Liposomes encapsulating compounds of the present disclosure contain at least one or more phospholipids, such as phosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, and sphingomyelin.

[0107] The fatty acid residue in the phospholipid is not particularly limited, and examples thereof include saturated or unsaturated fatty acid residues having 14 to 18 carbon atoms, and specific examples include acyl groups derived from fatty acids such as myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, etc. Furthermore, phospholipids derived from natural products such as egg yolk lecithin and soybean lecithin, as well as hydrogenated egg yolk lecithin and hydrogenated soybean lecithin (also called hydrogenated soybean phospholipid or hydrogenated soybean phosphatidylcholine) in which the unsaturated fatty acid residues of these phospholipids have been hydrogenated, can also be used.

[0108] The amount (molar fraction) of phospholipids to be mixed with respect to the total liposome membrane components is not particularly limited, but is preferably 30 to 80%, more preferably 40 to 70%.

[0109] Liposomes encapsulating the compounds of the present disclosure may contain sterols. Examples of sterols include cholesterol, β-sitosterol, stigmasterol, campesterol, brassica sterol, ergosterol, and fucosterol. Cholesterol is a preferred example of the sterol. The amount (molar fraction) of sterols relative to the total liposome membrane components is not particularly limited, but is preferably 0 to 60%, more preferably 10 to 50%, and even more preferably 30 to 50%.

[0110] Liposomes encapsulating the compounds of the present disclosure may contain polymer-modified lipids to improve in vivo retention. The amount (molar fraction) of the polymer-modified lipid relative to the total liposome membrane components is not particularly limited, but is preferably 0-20%, more preferably 1-10%. The polymer moiety of the polymer-modified lipid is preferably a hydrophilic polymer, more preferably a hydrophilic polymer in which the end of the polymer not bound to the lipid is alkoxylated. Specific examples of the polymer moiety of the polymer-modified lipid include, but are not limited to, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, methoxypolyethylene glycol, methoxypolypropylene glycol, methoxypolyvinyl alcohol, methoxypolyvinylpyrrolidone, ethoxypolyethylene glycol, ethoxypolypropylene glycol, ethoxypolyvinyl alcohol, ethoxypolyvinylpyrrolidone, propoxypolyethylene glycol, propoxypolypropylene glycol, propoxypolyvinyl alcohol, and propoxypolyvinylpyrrolidone. The polymer portion of the polymer-modified lipid preferably includes polyethylene glycol, methoxypolyethylene glycol, methoxypolypropylene glycol, ethoxypolyethylene glycol, ethoxypolypropylene glycol, propoxypolyethylene glycol, and propoxypolypropylene glycol.The polymer portion of the polymer-modified lipid more preferably includes polyethylene glycol, methoxypolyethylene glycol, ethoxypolyethylene glycol, ethoxypolypropylene glycol, and propoxypolyethylene glycol.The polymer portion of the polymer-modified lipid is even more preferably includes polyethylene glycol and methoxypolyethylene glycol.The polymer portion of the polymer-modified lipid is most preferably includes methoxypolyethylene glycol.The molecular weight of the polymer portion of the polymer-modified lipid is not particularly limited, but may be, for example, 100 to 10,000 daltons, preferably 1,000 to 7,000 daltons, more preferably 1,500 to 5,000 daltons, and most preferably 1,500 to 3,000 daltons.

[0111] The lipid moiety of the polymer-modified lipid is not particularly limited, and examples thereof include phosphatidylethanolamine and diacylglycerol. Preferred examples of the lipid moiety of the polymer-modified lipid include phosphatidylethanolamine having a saturated or unsaturated fatty acid residue having 14 to 18 carbon atoms, and diacylglycerol having a saturated or unsaturated fatty acid residue having 14 to 18 carbon atoms, more preferred examples include phosphatidylethanolamine having a saturated fatty acid residue having 14 to 18 carbon atoms, and diacylglycerol having a saturated fatty acid residue having 14 to 18 carbon atoms, and even more preferred examples include phosphatidylethanolamine having a palmitoyl group or a stearoyl group, and diacylglycerol having a palmitoyl group or a stearoyl group. The most preferred example of the lipid moiety of the polymer-modified lipid is distearoylphosphatidylethanolamine.

[0112] Liposomes encapsulating the compounds of the present disclosure can contain pharmaceutically acceptable additives, such as inorganic acids, inorganic acid salts, organic acids, organic acid salts, sugars, buffers, antioxidants, and polymers.

[0113] Inorganic acids include, for example, phosphoric acid, hydrochloric acid, and sulfuric acid.

[0114] Inorganic acid salts include, for example, sodium hydrogen phosphate, sodium chloride, ammonium sulfate, and magnesium sulfate.

[0115] Organic acids include, for example, citric acid, acetic acid, succinic acid, and tartaric acid.

[0116] Organic acid salts include, for example, sodium citrate, sodium acetate, disodium succinate, and sodium tartrate.

[0117] Sugars include, for example, glucose, sucrose, mannitol, sorbitol, and trehalose.

[0118] Examples of buffering agents include L-arginine, L-histidine, trometamol (trishydroxymethylaminomethane, Tris) and salts thereof.

[0119] Antioxidants include, for example, sodium sulfite, L-cysteine, sodium thioglycolate, sodium thiosulfate, ascorbic acid, and tocopherol.

[0120] Polymers include, for example, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, and sodium carboxymethylcellulose.

[0121]

[0039] Below, examples of methods for producing the compounds of the present disclosure represented by formula (1) will be described, but the methods for producing the compounds of the present disclosure are not limited to these. The compounds used in the following production methods may form salts as long as they do not interfere with the reaction.

[0122] The compounds of the present disclosure can be produced using known compounds as starting materials, for example, by the following production methods A, B, C, D, E, F, G, H, I, J, K, L, M, N, O, P, Q, and R, or methods similar thereto, or by appropriately combining synthesis methods known to those skilled in the art. Compounds of the present disclosure other than formula (a2) can also be produced by appropriately combining methods similar thereto or synthesis methods known to those skilled in the art.

[0123] Production Method A The compound of the present disclosure represented by formula (a2) can be produced, for example, by the following method. [In the formula, R １ , R ２ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group. １Examples of the protecting groups include the amino protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0124] [Step 1] Compound (a2) can be prepared by removing the protecting group P of compound (a1) obtained by the following production method. １ This step can be carried out, for example, in accordance with the method described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0125] Production Method B The compound of the present disclosure represented by formula (a1) can be produced, for example, by the following method. [In the formula, R １ , R ２ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group, and P ２ means a protecting group for phenol. １ Examples of the protecting groups include the amino protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999). ２ Examples of the protecting groups include the phenol protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0126] Compound (b3) is commercially available.

[0127] [Step 1] Compound (b2) can be prepared by removing the protecting group P of compound (b1) obtained by the following production method. ２ This step can be carried out, for example, in accordance with the method described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0128] [Step 2] Compound (a1) can be produced by subjecting compound (b2) and compound (b3) to Mitsunobu reaction in an appropriate solvent in the presence of a Mitsunobu reagent.

[0129] Examples of Mitsunobu reagents include diethyl azodicarboxylate (DEAD), isopropyl azodicarboxylate (DIAD), N,N,N',N'-tetraisopropylazodicarboxamide (TIPA), 1,1'-(azodicarbonyl)dipiperidine (ADDP), N,N,N',N'-tetramethylazodicarboxamide (TMAD), triphenylphosphine, tributylphosphine, and the like. Cyanomethylenetrimethylphosphorane (CMMP) and cyanomethylenetributylphosphorane (CMBP) can also be used.

[0130] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, anisole, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; or mixed solvents thereof, etc. Preferred examples of the solvent include toluene, benzene, THF, 1,4-dioxane, and mixed solvents thereof.

[0131] The reaction time is usually 5 minutes to 72 hours, preferably 12 to 24 hours.

[0132] The reaction temperature is usually 0°C to 100°C, preferably 0°C to 50°C.

[0133] Production Method C The compound of the present disclosure represented by formula (b1) can be produced, for example, by the following method. [In the formula, R １ , R ２ , X, Y and Z are as defined in item 1, and P ２ means a protecting group for phenol. ２ Examples of the protecting groups include the phenol protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0134] [Step 1] Compound (c2) can be produced by reacting compound (c1) with hydrazine monohydrate in a suitable solvent in the presence or absence of a suitable acid.

[0135] Examples of the acid include acetic acid, propionic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, hydrochloric acid, sulfuric acid, camphorsulfonic acid, etc. Preferred examples of the acid include acetic acid and p-toluenesulfonic acid.

[0136] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, anisole, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; alcohol solvents such as methanol, ethanol, isopropyl alcohol, butanol, etc.; or mixtures thereof. Preferred examples of the solvent include 1,4-dioxane, toluene, ethanol, etc.

[0137] The reaction time is usually 5 minutes to 72 hours, preferably 12 to 24 hours.

[0138] The reaction temperature is usually 0°C to 200°C, preferably 50°C to 100°C.

[0139] [Step 2] Compound (b1) can be produced by reacting compound (c2) with a 5-chloropyrazine derivative in the presence of a suitable base in a suitable solvent. Compound (b1) can also be produced by reacting compound (c2) with a 5-chloropyrazine derivative in the presence of a suitable palladium catalyst, ligand, and base in a suitable solvent.

[0140] Examples of the base include organic bases such as triethylamine, diisopropylethylamine, tributylamine, 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), pyridine, dimethylaminopyridine, picoline, N-methylmorpholine (NMM), and N-ethylmorpholine, and inorganic bases such as sodium hydrogencarbonate, potassium hydrogencarbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, and potassium hydroxide. Preferred examples of the base include triethylamine, diisopropylethylamine, N-ethylmorpholine, potassium carbonate, and cesium carbonate.

[0141] Examples of the palladium catalyst include salts such as palladium chloride and palladium acetate, and zero-valent palladium complexes such as tris(dibenzylideneacetone)dipalladium(0) chloroform adduct.

[0142] Examples of the ligand include phosphine ligands such as 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (Xantphos), 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (SPhos), and 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (XPhos). A preferred example of the ligand is 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (Xantphos).

[0143] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, anisole, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; or mixtures thereof. Preferred examples of the solvent include tetrahydrofuran, 1,4-dioxane, toluene, dimethyl sulfoxide, etc.

[0144] The reaction time is usually 5 minutes to 48 hours, preferably 1 hour to 6 hours.

[0145] The reaction temperature is usually 0°C to 200°C, preferably 50°C to 100°C.

[0146] Production Method D The compound of the present disclosure represented by formula (c1) can be produced, for example, by the following method.

[0147] [In the formula, R１ , X, Y and Z are as defined in item 1, and P ２ means a protecting group for phenol. ２ Examples of the protecting groups include the phenol protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0148] [Step 1] Compound (c1) can be produced by reacting compound (d1) with acetonitrile in the presence of a suitable base in a suitable solvent.

[0149] As the base, an inorganic base can be used. Examples of the inorganic base include hydrides such as sodium hydride, alkali halides such as potassium fluoride, alkali hydroxides such as sodium hydroxide and potassium hydroxide, alkali carbonates such as sodium carbonate, potassium carbonate, cesium carbonate and sodium bicarbonate, alkali alkoxides such as sodium ethoxide, sodium tert-butoxide and potassium tert-butoxide, and alkali metals such as n-butyllithium, methyllithium and isopropylmagnesium bromide. Preferred examples of the base include sodium ethoxide, sodium tert-butoxide, potassium tert-butoxide and n-butyllithium.

[0150] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, anisole, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; alcohol solvents such as methanol, ethanol, isopropyl alcohol, butanol, etc.; and mixtures thereof. Preferred solvents include tetrahydrofuran, toluene, etc.

[0151] The reaction time is usually 5 minutes to 72 hours, preferably 12 to 24 hours, and the reaction temperature is usually 0°C to 200°C, preferably 50°C to 100°C.

[0152] Production Method E The compound of the present disclosure represented by formula (b1) can be produced, for example, by the following method. [In the formula, R １ , R ２ , X, Y and Z are as defined in item 1, and P ２ means a protecting group for phenol. ２ Examples of the protecting groups include the phenol protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0153] [Step 1] Compound (e2) can be produced by reacting compound (e1) with carbon disulfide and iodomethane in the presence of a suitable base in a suitable solvent.

[0154] The base may be an inorganic base. Examples of the inorganic base include hydrides such as sodium hydride, alkali halides such as potassium fluoride, alkali hydroxides such as sodium hydroxide and potassium hydroxide, alkali carbonates such as sodium carbonate, potassium carbonate, cesium carbonate and sodium hydrogencarbonate, alkali alkoxides such as sodium ethoxide, sodium tert-butoxide and potassium tert-butoxide, and alkali metals such as n-butyllithium, methyllithium and isopropylmagnesium bromide. Preferred examples of the base include sodium hydride and potassium tert-butoxide.

[0155] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, anisole, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; alcohol solvents such as methanol, ethanol, isopropyl alcohol, butanol, etc.; and mixtures thereof. Preferred examples of the solvent include tetrahydrofuran, toluene, N,N-dimethylformamide, etc.

[0156] The reaction time is usually 5 minutes to 72 hours, preferably 30 minutes to 2 hours.

[0157] The reaction temperature is usually from -78°C to 200°C, preferably from 0°C to 25°C.

[0158] [Step 2] Compound (e3) can be produced by reacting compound (e2) with a 5-aminopyrazine derivative in the presence of a suitable base in a suitable solvent.

[0159] Examples of the base include organic bases such as triethylamine, diisopropylethylamine, tributylamine, 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), pyridine, dimethylaminopyridine, picoline, N-methylmorpholine (NMM), and N-ethylmorpholine, and inorganic bases such as sodium hydrogencarbonate, potassium hydrogencarbonate, sodium carbonate, potassium carbonate, sodium hydroxide, and potassium hydroxide. Preferred examples of the base include triethylamine and diisopropylethylamine.

[0160] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, anisole, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; or mixtures thereof. Preferred examples of the solvent include tetrahydrofuran, toluene, dimethyl sulfoxide, etc.

[0161] The reaction time is usually 5 minutes to 48 hours, preferably 2 to 8 hours.

[0162] The reaction temperature is usually 0°C to 200°C, preferably 50°C to 100°C.

[0163] [Step 3] Compound (b1) can be produced by reacting compound (e3) with hydrazine monohydrate in a suitable solvent in the presence or absence of a suitable acid.

[0164] Examples of the acid include acetic acid, propionic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, hydrochloric acid, sulfuric acid, camphorsulfonic acid, etc. Preferred examples of the acid include acetic acid.

[0165] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, anisole, and 1,4-dioxane; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, and xylene; ester solvents such as ethyl acetate and methyl acetate; aprotic solvents such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, and dimethyl sulfoxide; alcohol solvents such as methanol, ethanol, isopropyl alcohol, and butanol; or mixtures thereof. Preferred solvents include 1,4-dioxane and ethanol.

[0166] The reaction time is usually 5 minutes to 72 hours, preferably 12 to 24 hours.

[0167] The reaction temperature is usually 0°C to 200°C, preferably 50°C to 100°C.

[0168] Production Method F The compound of the present disclosure represented by formula (c1) can be produced, for example, by the following method. [In the formula, R １ , X, Y and Z are as defined in item 1, and P ２ means a protecting group for phenol. ２ Examples of the protecting groups include the phenol protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0169] [Step 1] Compound (f1) can be produced by reacting compound (e1) with dimethylformamide dimethyl acetal in an appropriate solvent.

[0170] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, anisole, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; or mixtures thereof. Preferred examples of the solvent include N,N-dimethylformamide, etc.

[0171] The reaction time is usually 5 minutes to 48 hours, preferably 24 to 48 hours.

[0172] The reaction temperature is usually 0°C to 200°C, preferably 60°C to 120°C.

[0173] [Step 2] Compound (f2) can be produced by reacting compound (f1) with hydroxyamine hydrochloride in an appropriate solvent.

[0174] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, anisole, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; alcohol solvents such as methanol, ethanol, isopropyl alcohol, butanol, etc.; or mixtures thereof. Preferred examples of the solvent include ethanol and isopropyl alcohol.

[0175] The reaction time is usually 5 minutes to 48 hours, preferably 6 to 12 hours.

[0176] The reaction temperature is usually 0°C to 200°C, preferably 40°C to 80°C.

[0177] [Step 3] Compound (c1) can be produced by reacting compound (f2) with a base in an appropriate solvent.

[0178] Examples of the base include inorganic bases such as sodium hydrogen carbonate, potassium hydrogen carbonate, sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, etc. Preferred examples of the base include sodium hydroxide and potassium hydroxide.

[0179] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, anisole, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; protic solvents such as water, methanol, ethanol, isopropyl alcohol, butanol, etc.; or mixtures thereof. Preferred examples of the solvent include a mixed solvent of water and ethanol.

[0180] The reaction time is usually 5 minutes to 48 hours, preferably 1 hour to 6 hours.

[0181] The reaction temperature is usually 0°C to 200°C, preferably 20°C to 50°C.

[0182] Production Method G The compound of the present disclosure represented by formula (a1) can be produced, for example, by the following method. [In the formula, R １ , R ２ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group. １Examples of the protecting groups include the amino protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0183] [Step 1] Compound (g2) can be produced from compound (g1) obtained by the following production method by the method described in Step 1 of Production Method C or a method analogous thereto.

[0184] [Step 2] Compound (a1) can be produced from compound (g2) by the method described in Step 2 of Production Method C or a method analogous thereto.

[0185] Production Method H The compound represented by formula (a1) can be produced, for example, by the following method. [In the formula, R １ , R ２ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group. １ Examples of the protecting groups include the amino protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0186] [Step 1] Compound (h2) can be produced from compound (h1) obtained by the following production method by the method described in Step 1 of Production Method E or a method analogous thereto.

[0187] [Step 2] Compound (h3) can be produced from compound (h2) by the method described in step 2 of production method E or a method analogous thereto. [Step 3] Compound (a1) can be produced from compound (h3) by the method described in step 3 of production method E or a method analogous thereto.

[0188] Production Method I The compound represented by formula (g1) can be produced, for example, by the following method. [In the formula, R １ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group. １ Examples of the protecting groups include the amino protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0189] [Step 1] Compound (i1) can be produced from compound (h1) obtained by the following production method by the method described in Step 1 of Production Method F or a method analogous thereto.

[0190] [Step 2] Compound (i2) can be produced from compound (i1) by the method described in Step 2 of Production Method F or a method analogous thereto.

[0191] [Step 3] Compound (g1) can be produced from compound (i2) by the method described in Step 3 of Production Method F or a method analogous thereto.

[0192] Production Method J The compound represented by formula (g1) can also be produced, for example, by the following method. [In the formula, R １ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group. １ Examples of the protecting groups include the amino protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0193] [Step 1] Compound (g1) can be produced from compound (j1) obtained by the following production method by the method described in Step 1 of Production Method D or a method analogous thereto.

[0194] Production Method K The compound of the present disclosure represented by formula (d1) can be produced, for example, by the following method. [In the formula, R １ , X, Y and Z are as defined in item 1, and P ２ means a protecting group for phenol. ２ Examples of the protecting groups include the phenol protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0195] The compound (k1) is commercially available.

[0196] [Step 1] Compound (d1) can be produced from compound (k1) by, for example, the method described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0197] Production Method L The compound of the present disclosure represented by formula (e1) can be produced, for example, by the following method. [In the formula, R １ , X, Y and Z are as defined in item 1, and P ２ means a protecting group for phenol. ２ Examples of the protecting groups include the phenol protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0198] Compound (l1) is commercially available.

[0199] [Step 1] Compound (e1) can be produced from compound (l1) by the method described in Step 1 of Production Method K or a method analogous thereto.

[0200] Production Method M The compound of the present disclosure represented by formula (j1) can be produced, for example, by the following method. [In the formula, R １ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group. １ Examples of the protecting groups include the amino protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0201] The compound (k1) and the compound (b3) are commercially available.

[0202] [Step 1] Compound (j1) can be produced from compound (k1) and compound (b3) by the method described in step 2 of production method B or a method analogous thereto.

[0203] Production Method N The compound of the present disclosure represented by formula (j1) can be produced, for example, by the following method. [In the formula, R １ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group, and LG means a leaving group. １ Examples of LG include the amino-protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999). Examples of LG include halogen, methanesulfonyloxy, p-toluenesulfonyloxy, and trifluoromethanesulfonyloxy.]

[0204] The compound (k1) and the compound (n1) are commercially available.

[0205] [Step 1] Compound (j1) can be produced by reacting compound (k1) with compound (n1) in the presence or absence of a suitable base in a suitable solvent.

[0206] Examples of the base include organic bases such as triethylamine, diisopropylethylamine, tributylamine, 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), pyridine, 4-dimethylaminopyridine, picoline, N-methylmorpholine (NMM), etc., and inorganic bases such as sodium hydrogen carbonate, potassium hydrogen carbonate, sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, etc. Preferred examples of the base include triethylamine, diisopropylethylamine, potassium carbonate, sodium hydroxide, etc.

[0207] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include alcoholic solvents such as methanol, ethanol, 2-propanol (isopropyl alcohol), tert-butanol, etc.; etheric solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, anisole, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; or mixtures thereof. Preferred examples of the solvent include 2-propanol, tetrahydrofuran, toluene, acetonitrile, N,N-dimethylformamide, etc.

[0208] The reaction temperature is usually from -80°C to reflux, preferably from 25°C to 90°C.

[0209] The reaction time is usually 30 minutes to 48 hours, preferably 6 to 12 hours.

[0210] Production Method O The compound of the present disclosure represented by formula (h1) can be produced, for example, by the following method. [In the formula, R １ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group. １ Examples of the protecting groups include the amino protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999).

[0211] The compound (l1) and the compound (b3) are commercially available.

[0212] [Step 1] Compound (h1) can be produced from compound (l1) and compound (b3) by the method described in step 2 of production method B or a method analogous thereto.

[0213] Production Method P The compound of the present disclosure represented by formula (h1) can be produced, for example, by the following method. [In the formula, R １ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group, and LG means a leaving group. １ Examples of LG include the amino-protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999). Examples of LG include halogen, methanesulfonyloxy, p-toluenesulfonyloxy, and trifluoromethanesulfonyloxy.]

[0214] Compound (l1) and compound (n1) are commercially available.

[0215] [Step 1] Compound (h1) can be produced from compound (l1) and compound (n1) by the method described in step 2 of production method B or a method analogous thereto.

[0216] Production Method Q The compound of the present disclosure represented by formula (h1) can be produced, for example, by the following method. [In the formula, R １ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group, and LG means a leaving group. １ Examples of LG include the amino-protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999). Examples of LG include halogen, methanesulfonyloxy, p-toluenesulfonyloxy, and trifluoromethanesulfonyloxy.]

[0217] The compound (q1), the compound (q2) and the compound (b3) are commercially available.

[0218] [Step 1] Compound (q3) can be produced from compound (q1) and compound (q2) by the method described in step 1 of production method N or a method analogous thereto.

[0219] [Step 2] Compound (q4) can be produced by reacting compound (q3) with acetaldehyde in the presence of a suitable base in a suitable solvent.

[0220] Examples of the base include n-butyllithium, sec-butyllithium, tert-butyllithium, lithium diisopropylamide, lithium tetramethylpiperidide, lithium hexamethyldisilazide, sodium hexamethyldisilazide, potassium hexamethyldisilazide, isopropylmagnesium chloride-lithium chloride complex, and the like.

[0221] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, anisole, xylene, etc.; halogenated solvents such as methylene chloride, chloroform, 1,2-dichloroethane, etc.; aprotic solvents such as acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; or mixtures thereof. Tetrahydrofuran is a preferred solvent.

[0222] The reaction temperature is usually from -80°C to reflux, preferably from -80°C to 25°C.

[0223] The reaction time is usually 30 minutes to 48 hours, preferably 6 to 12 hours.

[0224] [Step 3] Compound (q5) can be produced by reacting compound (q4) with a suitable oxidizing agent in the presence or absence of a suitable base in a suitable solvent.

[0225] Examples of the oxidizing agent include manganese dioxide, Dess-Martin reagent, dimethyl sulfoxide-oxalyl chloride, dimethyl sulfoxide-trifluoroacetic anhydride, sulfur trioxide-pyridine complex, 2,2,6,6-tetramethylpiperidine-1-oxy radical-sodium hypochlorite, etc. Examples of the base include organic bases such as triethylamine, diisopropylethylamine, tributylamine, 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), pyridine, 4-dimethylaminopyridine, picoline, N-methylmorpholine (NMM), etc., and inorganic bases such as sodium hydrogen carbonate, potassium hydrogen carbonate, sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, etc.

[0226] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include halogenated solvents such as methylene chloride, chloroform, 1,2-dichloroethane, etc.; ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, anisole, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; or mixtures thereof. Preferred examples of the solvent include methylene chloride, tetrahydrofuran, toluene, acetonitrile, N,N-dimethylformamide, etc.

[0227] The reaction temperature is usually −80° C. to reflux, preferably −80° C. to 25° C. The reaction time is usually 30 minutes to 48 hours, preferably 6 to 12 hours.

[0228] [Step 4] Compound (h1) can be produced from compound (q5) and compound (b3) by the method described in step 2 of production method B or a method analogous thereto.

[0229] Production Method R The compound of the present disclosure represented by formula (i2) can be produced, for example, by the following method. [In the formula, R １ , R １４ , X, Y, Z, L, V and W are as defined in item 1; １ means an amino protecting group, and LG means a leaving group. １ Examples of LG include the amino-protecting groups described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, John Wiley & Sons, Inc., 1999). Examples of LG include halogen, methanesulfonyloxy, p-toluenesulfonyloxy, and trifluoromethanesulfonyloxy.]

[0230] The compound (l1) and the compound (b3) are commercially available.

[0231] [Step 1] Compound (r1) can be produced from compound (l1) by the method described in Step 1 of Production Method F or a method analogous thereto.

[0232] [Step 2] Compound (r2) can be produced from compound (r1) by the method described in Step 2 of Production Method F or a method analogous thereto.

[0233] [Step 3] Compound (r3) can be produced by reacting compound (r2) with an appropriate halogenating agent or sulfonylating agent in the presence or absence of a suitable base, in an appropriate solvent or without a solvent.

[0234] Examples of the halogenating agent or sulfonylating agent include thionyl chloride, phosphorus oxychloride, oxalyl chloride, phosphorus tribromide, methanesulfonyl chloride, N-phenylbis(trifluoromethanesulfonimide), trifluoromethanesulfonic anhydride, and the like.

[0235] Examples of the base include organic bases such as triethylamine, diisopropylethylamine, tributylamine, 1,5-diazabicyclo[4.3.0]non-5-ene (DBN), 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), pyridine, 4-dimethylaminopyridine, picoline, and N-methylmorpholine (NMM); and inorganic bases such as sodium hydrogen carbonate, potassium hydrogen carbonate, sodium carbonate, potassium carbonate, sodium hydroxide, and potassium hydroxide.

[0236] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include halogenated solvents such as methylene chloride, chloroform, 1,2-dichloroethane, etc.; ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, anisole, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; or mixtures thereof. Preferred examples of the solvent include methylene chloride, tetrahydrofuran, toluene, acetonitrile, N,N-dimethylformamide, etc.

[0237] The reaction temperature is usually from −80° C. to reflux, preferably from 0° C. to reflux.

[0238] The reaction time is usually 30 minutes to 48 hours, preferably 30 minutes to 12 hours.

[0239] [Step 4] Compound (i2) can be produced by reacting compound (r3) with compound (b3) in the presence of a suitable base in a suitable solvent, catalyzed by a suitable palladium source and ligand.

[0240] Examples of the base include organic bases such as triethylamine, diisopropylethylamine, tributylamine, pyridine, 4-dimethylaminopyridine, picoline, N-methylmorpholine (NMM), etc., and inorganic bases such as sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide, etc. Preferred examples of the base include diisopropylethylamine, cesium carbonate, etc.

[0241] Examples of palladium sources include salts such as palladium chloride and palladium acetate, and zero-valent palladium complexes such as tris(dibenzylideneacetone)dipalladium(0) chloroform adduct. Examples of ligands include phosphine ligands such as 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (Xantphos), 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (SPhos), and 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (XPhos). A preferred ligand is 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (Xantphos).

[0242] The solvent is not particularly limited as long as it is inert under the reaction conditions of this step, and examples thereof include halogenated solvents such as methylene chloride, chloroform, 1,2-dichloroethane, etc.; ether solvents such as diethyl ether, diisopropyl ether, tetrahydrofuran, methylcyclopentyl ether, 1,4-dioxane, etc.; aromatic hydrocarbon solvents such as benzene, toluene, chlorobenzene, anisole, xylene, etc.; ester solvents such as ethyl acetate, methyl acetate, etc.; aprotic solvents such as acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-2-pyrrolidinone, 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide, etc.; or mixtures thereof. Preferred examples of the solvent include tetrahydrofuran, 1,4-dioxane, toluene, acetonitrile, N,N-dimethylformamide, etc.

[0243] The reaction temperature is usually from 0° C. to reflux, preferably from 50° C. to reflux. The reaction time is usually from 5 minutes to 72 hours, preferably from 2 to 12 hours.

[0244] In the above-described production methods, starting materials or intermediates for which production methods are not described are either commercially available or can be synthesized from commercially available compounds by methods known to those skilled in the art or methods similar thereto.

[0245] In each reaction of the production method described above, even when the use of a protecting group is not specifically specified, a protecting group can be used as needed. For example, when any functional group other than the reactive site changes under the reaction conditions described, or when the method described is inappropriate to carry out without a protecting group, the target compound can be obtained by protecting the functional group other than the reactive site as needed and deprotecting it after the reaction or after a series of reactions.

[0246] Examples of the protecting group that can be used include those described in Protective Groups in Organic Synthesis (Theodora W. Greene, Peter G. M. Wuts, published by John Wiley & Sons, Inc., 1999). Specific examples of the amino protecting group include benzyloxycarbonyl, tert-butoxycarbonyl, acetyl, benzyl, etc. Specific examples of the hydroxyl protecting group include trialkylsilyl such as trimethylsilyl and tert-butyldimethylsilyl, acetyl, benzyl, etc.

[0247] Introduction and removal of the protecting group can be carried out by a method commonly used in organic synthetic chemistry (for example, see the aforementioned Protective Groups in Organic Synthesis) or a method analogous thereto.

[0248] In this specification, protecting groups, condensing agents, etc. may be represented by abbreviations according to IUPAC-IUB (International Board of Biochemical Nomenclature) commonly used in this technical field. Note that the names of compounds used in this specification do not necessarily conform to the IUPAC nomenclature.

[0249] The intermediates or target compounds in the above-described production methods can be converted into other compounds included in the present disclosure by appropriately converting their functional groups (for example, by protecting or deprotecting the functional groups as necessary, and then performing various conversions using amino, hydroxyl, carbonyl, halogen, etc. as a stepping stone). The conversion of functional groups can be performed by a commonly used method (see, for example, Comprehensive Organic Transformations, R. C. Larock, John Wiley & Sons Inc. (1999)).

[0250] The intermediates and target compounds in the above-described production methods can be isolated and purified by purification methods commonly used in organic synthetic chemistry (e.g., neutralization, filtration, extraction, washing, drying, concentration, recrystallization, various types of chromatography, etc.). In addition, the intermediates can also be used in the next reaction without any particular purification.

[0251] "Pharmaceutically acceptable salts" include acid addition salts and base addition salts. For example, acid addition salts include inorganic acid salts such as hydrochloride, hydrobromide, sulfate, hydroiodide, nitrate, and phosphate, and organic acid salts such as citrate, oxalate, phthalate, fumarate, maleate, succinate, malate, acetate, formate, propionate, benzoate, trifluoroacetate, methanesulfonate, benzenesulfonate, para-toluenesulfonate, and camphorsulfonate. Furthermore, base addition salts include inorganic base salts such as sodium salt, potassium salt, calcium salt, magnesium salt, barium salt, and aluminum salt, and organic base salts such as trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, tromethamine [tris(hydroxymethyl)methylamine], tert-butylamine, cyclohexylamine, dicyclohexylamine, and N,N-dibenzylethylamine. Furthermore, "pharmaceutically acceptable salts" also include amino acid salts with basic or acidic amino acids such as arginine, lysine, ornithine, aspartic acid, or glutamic acid.

[0252] Suitable salts of starting materials and intermediates, as well as salts acceptable as pharmaceutical raw materials, are conventional non-toxic salts, including, for example, acid addition salts such as organic acid salts (e.g., acetate, trifluoroacetate, maleate, fumarate, citrate, tartrate, methanesulfonate, benzenesulfonate, formate, toluenesulfonate, etc.) and inorganic acid salts (e.g., hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate, etc.), salts with amino acids (e.g., arginine, aspartic acid, glutamic acid, etc.), metal salts such as alkali metal salts (e.g., sodium salt, potassium salt), alkaline earth metal salts (e.g., calcium salt, magnesium salt), ammonium salts, organic base salts (e.g., trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, etc.), and others that can be appropriately selected by those skilled in the art.

[0253] In the present disclosure, any one or more of the compounds represented by formulas (1) to (5) １ H ２ Deuterium-converted products converted to H(D) are also included in the compounds represented by formulas (1) to (5).

[0254] The present disclosure includes compounds represented by formulas (1) to (5) or pharmaceutically acceptable salts thereof. In addition, the compounds of the present disclosure may exist in the form of hydrates and / or solvates with various solvents (such as ethanol solvates), and these hydrates and / or solvates are also included in the compounds of the present disclosure.

[0255] Furthermore, the compounds of the present disclosure include all possible isomers such as optical isomers based on optically active centers, atropisomers based on axial or planar chirality resulting from restricted intramolecular rotation, other stereoisomers, tautomers, geometric isomers, and the like, as well as all types of crystalline forms, and mixtures thereof.

[0256] In particular, optical isomers and atropisomers can be obtained as racemates, or as optically active isomers when optically active starting materials or intermediates are used. Furthermore, if necessary, at an appropriate stage of the production method, the racemate of the corresponding starting material, intermediate, or final product can be physically or chemically resolved into its optical antipodes by known separation methods such as a method using an optically active column or fractional crystallization. Examples of such resolution methods include a diastereomeric method in which a racemate is reacted with an optically active resolving agent to synthesize two diastereomers, which are then separated by a method such as fractional crystallization, taking advantage of their different physical properties.

[0257] When it is desired to obtain a pharmaceutically acceptable salt of a compound of the present disclosure, if the compound represented by formulas (1) to (5) is obtained in the form of a pharmaceutically acceptable salt, it may be purified as is, or if it is obtained in the free form, it may be dissolved or suspended in an appropriate organic solvent, and an acid or a base may be added to form a salt by a conventional method.

[0258] Liposomes encapsulating the compounds of the present disclosure can be produced, for example, by the following method.

[0259] [Step 1] Membrane components such as phospholipids and cholesterol are dissolved in an organic solvent such as chloroform, and the organic solvent is evaporated in a flask to form a thin film of the lipid mixture on the inner wall of the flask. Alternatively, the lipid mixture may be obtained as a lyophilized product by dissolving the components in t-butyl alcohol or the like and then lyophilizing the solution. Alternatively, membrane components such as phospholipids and cholesterol may be dissolved in an organic solvent and then powdered using a CRUX instant vacuum dryer (manufactured by Hosokawa Micron Corporation) to obtain a lipid mixture, which is available from Nippon Fine Chemical Co., Ltd. under the name Presome (registered trademark).

[0260] [Step 2] An internal aqueous phase solution such as an aqueous ammonium sulfate solution is added to the lipid mixture obtained in step 1 and dispersed to obtain a crude liposome dispersion.

[0261] [Step 3] The crude liposome dispersion obtained in step 2 is passed through a filter using an extruder to obtain a desired particle size. Alternatively, the crude liposome dispersion obtained in step 2 is extruded from a nozzle at high pressure using a high-pressure homogenizer to obtain a desired particle size. The liposome particle size is not particularly limited, but is, for example, 10 nm to 200 nm, preferably 30 nm to 150 nm, more preferably 40 nm to 140 nm, even more preferably 50 to 120 nm, and most preferably 60 to 100 nm. The liposome particle size is an average value measured by dynamic light scattering, and can be measured, for example, using a Zetasizer Nano ZS (Malvern Instruments).

[0262] [Step 4] The external aqueous phase of the liposome solution obtained in step 3 is replaced by gel filtration, dialysis, tangential flow filtration, ultracentrifugation, or the like.

[0263] [Step 5] The liposome solution obtained in step 4, in which the external aqueous phase has been replaced, is incubated with the compound to be encapsulated, thereby encapsulating the compound in the liposomes.

[0264] [Step 6] The resulting liposomes encapsulating the compound are subjected to gel filtration, dialysis, tangential flow filtration, ultracentrifugation, etc. to remove any unencapsulated compound. Note that if a desired encapsulation rate is obtained in step 5, step 6 can be omitted.

[0265] The compounds of the present disclosure can be used in combination with other drugs to enhance their effects. Specifically, the compounds of the present disclosure can be used in combination with drugs such as hormone therapy agents, chemotherapy agents, immunotherapy agents, or drugs that inhibit cell growth factors and their receptor activity. Hereinafter, drugs that can be used in combination with the compounds of the present disclosure will be abbreviated as "concomitant drugs."

[0266] The compounds of the present disclosure exhibit excellent anticancer effects even when used as a single agent, but by further using them in combination with one or more of the above-mentioned concomitant drugs (multidrug combination), the effects can be further enhanced or the patient's QOL can be improved.

[0267] Examples of "hormonal therapeutic agents" include fosfestrol, diethylstilbestrol, chlorotrianisene, medroxyprogesterone acetate, megestrol acetate, chlormadinone acetate, cyproterone acetate, danazol, dienogest, asoprisnil, allylestrenol, gestrinone, nomegestol, tadenane, mepartricin, raloxifene, ormeloxifene, levormeloxifene, antiestrogens (e.g., tamoxifen citrate, toremifene citrate, etc.), birth control pills, mepitiostane, testololactone, aminoglutethimide, LH-RH derivatives (LH-RH agonists (e.g., goserelin acetate, buserelin, levothyroxine), levothyroxine, ... prorelin, etc.), LH-RH antagonists), droloxifene, epitiostanol, ethinyl estradiol sulfonate, aromatase inhibitors (e.g., fadrozole hydrochloride, anastrozole, letrozole, exemestane, vorozole, formestane, etc.), antiandrogens (e.g., flutamide, enzalutamide, apalutamide, bicalutamide, nilutamide, etc.), adrenocortical hormone drugs (e.g., dexamethasone, prednisolone, betamethasone, triamcinolone, etc.), androgen synthesis inhibitors (e.g., abiraterone, etc.), retinoids, and agents that slow the metabolism of retinoids (e.g., liarozole, etc.).

[0268] Examples of "chemotherapeutic agents" that can be used include alkylating agents, antimetabolites, anticancer antibiotics, plant-derived anticancer agents, molecular targeted therapeutic agents, immunomodulators, and other chemotherapeutic agents. Representative examples are listed below.

[0269] Examples of the "alkylating agent" include nitrogen mustard, nitrogen mustard-N-oxide hydrochloride, chlorambucil, cyclophosphamide, ifosfamide, thiotepa, carboquone, improsulfan tosylate, busulfan, nimustine hydrochloride, mitobronitol, melphalan, dacarbazine, ranimustine, estramustine sodium phosphate, triethylenemelamine, carmustine, lomustine, streptozotocin, Examples of drugs include benzodiazepine, pipobroman, etoglucide, carboplatin, cisplatin, miboplatin, nedaplatin, oxaliplatin, altretamine, ambamustine, dibrospidium hydrochloride, fotemustine, prednimustine, pumitepa, ribomustine, temozolomide, treosulfan, trofosfamide, zinostatin stimalamer, adozelesin, systostin, bizelesin, trabectedin, and DDS formulations thereof.

[0270] Examples of "antimetabolites" include mercaptopurine, 6-mercaptopurine riboside, thioinosine, methotrexate, pemetrexed, eocitabine, cytarabine, cytarabine ocfosfate, ancitabine hydrochloride, 5-FU drugs (e.g., fluorouracil, tegafur, UFT, doxifluridine, carmofur, galocitabine, emitefur, capecitabine, etc.), aminopterin, nelzarabine, leucoporin calcium, tabloid, butosin, folinate calcium, levofolinate calcium, cladribine, emitefur, fludarabine, gemcitabine, hydroxycarbamide, pentostatin, piritrexim, idoxuridine, mitoguazone, tiazofurin, ambamustine, bendamustine, and DDS preparations thereof.

[0271] Examples of "anticancer antibiotics" include actinomycin D, actinomycin C, mitomycin C, chromomycin A3, bleomycin hydrochloride, bleomycin sulfate, peplomycin sulfate, daunorubicin hydrochloride, doxorubicin hydrochloride, aclarubicin hydrochloride, pirarubicin hydrochloride, epirubicin hydrochloride, neocarzinostatin, mithramycin, sarkomycin, carzinophilin, mitotane, zorubicin hydrochloride, mitoxantrone hydrochloride, idarubicin hydrochloride, eribulin, and DDS preparations thereof.

[0272] Examples of "plant-derived anticancer agents" include etoposide, etoposide phosphate, vinblastine sulfate, vincristine sulfate, vindesine sulfate, teniposide, paclitaxel, docetaxel, DJ-927, vinorelbine, irinotecan, topotecan, and DDS formulations thereof.

[0273] Examples of "molecular targeted therapeutic agents" include imatinib, gefitinib, erlotinib, sorafenib, dasatinib, sunitinib, nilotinib, lapatinib, pazopanib, ruxolitinib, crizotinib, vemurafenib, vandetanib, ponatinib, cabozantinib, tofacitinib, regorafenib, bosutinib, axitinib, dabrafenib, trametinib, nintedanib, idelalisib, ceritinib, lenvatinib, palbociclib, alectinib, afatinib, osimertinib, ribociclib, and abemaciclib. , brigatinib, neratinib, copanlisib, cobimetinib, ibrutinib, acalabrutinib, encorafenib, binimetinib, baricitinib, fostamatinib, lorlatinib, erdafitinib, entrectinib, dacomitinib, sirolimus, everolimus, temsirolimus, olaparib, rucaparib, niraparib, venetoclax, azacitidine, decitabine, vorinostat, panobinostat, romidepsin, bortezomib, carfilzomib, larotrectinib, and ixazomib.

[0274] "Immunomodulators" include, for example, lenalidomide and pomalidomide.

[0275] Examples of "other chemotherapeutic agents" include sobuzoxane.

[0276] Examples of "immunotherapeutic agents (BRM)" include picibanil, krestin, sizofiran, lentinan, ubenimex, interferon, interleukin, macrophage colony-stimulating factor, granulocyte colony-stimulating factor, erythropoietin, lymphotoxin, BCG vaccine, Corynebacterium parvum, levamisole, polysaccharide K, procodazole, anti-CTLA4 antibody, anti-PD-1 antibody, anti-PD-L1 antibody, and Toll-like receptor agonists (e.g., TLR7 agonist, TLR8 agonist, TLR9 agonist, etc.).

[0277] The cell growth factor in the drug that inhibits the action of cell growth factors and their receptors may be any substance that promotes cell growth, and typically includes a peptide with a molecular weight of 20,000 or less that exerts its effect at low concentrations by binding to a receptor. Specifically, EGF (epidermal growth factor) or a substance having substantially the same activity as EGF (e.g., TGFalpha, etc.), insulin or a substance having substantially the same activity as insulin (e.g., insulin, IGF (insulin-like growth factor)-1, IGF-2, etc.), FGF (fibroblast growth factor) or a substance having substantially the same assay as FGF (e.g., acidic FGF, basic FGF, KGK (keratinocyte growth factor), FGF-10, etc.), and other cell growth factors (e.g., CSF (colony stimulating factor), EPO (erythropoietin), IL-2 (interleukin-2), NGF (nerve growth factor), etc.). growth factor), PDGF (platelet-derived growth factor), TGF-beta (transforming growth factor beta), HGF (hepatocyte growth factor), VEGF (vascular endothelial growth factor), heregulin, angiopoietin, etc.

[0278] The administration period of the compound of the present disclosure and the concomitant drug is not limited, and they may be administered to the subject simultaneously or at staggered times. The compound of the present disclosure and the concomitant drug may also be used as a combination drug. The dose of the concomitant drug can be appropriately selected based on clinically used doses. The compounding ratio of the compound of the present disclosure to the concomitant drug can be appropriately selected depending on the subject, administration route, target disease, symptoms, combination, etc. For example, when the subject is a human, 0.01 to 100 parts by weight of the concomitant drug may be used per 1 part by weight of the compound of the present disclosure. Furthermore, for the purpose of suppressing side effects, the compound of the present disclosure may be used in combination with drugs (concomitant drugs) such as antiemetics, hypnotics, and anticonvulsants.

[0279] "Exhibiting resistance to immune checkpoint inhibitors" means, in the case of solid cancers, a state in which tumor growth cannot be suppressed even when an immune checkpoint inhibitor is administered. "Exhibiting resistance to anti-PD-1 antibodies" means a state in which tumor growth cannot be suppressed even when the above-mentioned anti-PD-1 antibodies are administered. "Exhibiting resistance to anti-PD-L1 antibodies" means a state in which tumor growth cannot be suppressed even when the above-mentioned anti-PD-L1 antibodies are administered. In the case of blood cancers, it means a state in which an increase in the number of cancer cells in the blood cannot be suppressed even when an immune checkpoint inhibitor is administered. "Exhibiting resistance to anti-PD-1 antibodies" means a state in which an increase in the number of cancer cells in the blood cannot be suppressed even when the above-mentioned anti-PD-1 antibodies are administered. "Exhibiting resistance to anti-PD-L1 antibodies" means a state in which an increase in the number of cancer cells in the blood cannot be suppressed even when the above-mentioned anti-PD-L1 antibodies are administered.

[0280] Anti-PD-1 antibodies are proteins that recognize the PD-1 molecule, have a Y-shaped four-chain structure (two light and two heavy polypeptide chains), and are molecules that selectively recognize one molecule via the Fab region. Anti-PD-1 antibodies are produced by fusing antibody-producing B cells with myeloma to produce hybridomas, and then purifying the antibodies secreted into the culture supernatant.

[0281] The M1-M2 ratio of macrophages refers to the ratio of a cell population defined by M1 macrophage markers (e.g., CD86, STAT1, IL-1beta, iNOS, CXCL9, CXCL10, etc.) to a cell population defined by M2 macrophage markers (e.g., Mrc1, Arg1, CD163, etc.). A higher value indicates a more inflammatory environment within the tumor, making it easier for anti-PD-1 / PD-L1 antibodies to be effective.

[0282] A solid cancer with a high tumor mutation burden (Tumor Mutation Burden-High; TMB-High) refers to a cancer with a TMB score of 10 mut / Mb or more as measured in FoundationOne CDx Cancer Genomic Profile.

[0283] "Anti-PD-1 antibody responsive patients" refers to a state in which tumor growth is suppressed by administering anti-PD-1 antibodies. A state in which tumor growth is suppressed refers to stable disease (SD), partial response (PR), or complete response (CR) according to the RECIST criteria. In particular, a state in which the effect of tumor growth suppression continues for 6 months (24 weeks) or more is called durable clinical benefit (DCB).

[0284] The CHK1 inhibitor and the immune checkpoint inhibitor may be administered simultaneously, and then the anti-PD-1 antibody may be administered 3 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks later.

[0285] The M1-M2 ratio of macrophages of cancer patients showing resistance to immune checkpoint inhibitors is preferably 21 or less, 20 or less, 19 or less, 18 or less, 17 or less, 16 or less, 15 or less, 14 or less, 13 or less, 12 or less, 11 or less, 10 or less, 9.9 or less, 9.8 or less, 9.7 or less, 9.6 or less, 9.5 or less, 9.4 or less, 9.3 or less, 9.2 or less, 9.1 or less, 9.0 or less, 8.9 or less. 8.8 or less, 8.7 or less, 8.6 or less, 8.5 or less, 8.4 or less, 8.3 or less, 8.2 or less, 8.1 or less, 8.0 or less, 7.9 or less, 7.8 or less, 7.7 or less, 7.6 or less, 7.5 or less, 7. 4 or less, 7.3 or less, 7.2 or less, 7.1 or less, 7.0 or less, 6.9 or less, 6.8 or less, 6.7 or less, 6.6 or less, 6.5 or less, 6.4 or less, 6.3 or less, 6.2 or less, 6.1 or less, 6.0 or less, 5 .. 9 or less, 5.8 or less, 5.7 or less, 5.6 or less, 5.5 or less, 5.4 or less, 5.3 or less, 5.2 or less, 5.1 or less, 5.0 or less, 4.9 or less, 4.8 or less, 4.7 or less, 4.6 or less, 4.5 or less, 4.4 or less, 4.3 or less, 4.2 or less, 4.1 or less, 4.0 or less, 3.9 or less, 3.8 or less, 3.7 or less, 3.6 or less, 3.5 or less, 3.4 or less, 3.3 or less, 3.2 or less, 3.1 or less, 3.0 or less , 2.9 or less, 2.8 or less, 2.7 or less, 2.6 or less, 2.5 or less, 2.4 or less, 2.3 or less, 2.2 or less, 2.1 or less, 2.0 or less, 1.9 or less, 1.8 or less, 1.7 or less, 1.6 or less, 1.5 or less, 1.4 or less, 1.3 or less, 1.2 or less, 1.1 or less, 1.0 or less, 0.9 or less, 0.8 or less, 0.7 or less, 0.6 or less, 0.5 or less, 0.4 or less, 0.3 or less, 0.2 or less, 0.1 or less.

[0286] The number of CD8-positive cells in tumor tissue of a cancer patient exhibiting treatment resistance to immune checkpoint inhibitors is preferably 0.135 or less, 0.130 or less, 0.125 or less, 0.120 or less, 0.115 or less, 0.110 or less, 0.095 or less, 0.090 or less, 0.085 or less, 0.080 or less, 0.075 or less, 0.070 or less, 0.065 or less, 0.060 or less, 0.055 or less, 0.050 or less, 0.045 or less, 0.040 or less, 0.035 or less, 0.030 or less, 0.025 or less, 0.020 or less, 0.015 or less, or 0.010 or less.

[0287] In this specification, "or" is used when "at least one or more" of the items listed in the sentence can be employed. The same applies to "alternative." When specified in this specification as "within a range of two values," the range includes the two values ​​themselves.

[0288] All references cited herein, including scientific literature, patents, patent applications, and the like, are incorporated by reference in their entirety to the same extent as if each were specifically set forth.

[0289] The present disclosure has been described above by showing preferred embodiments for ease of understanding. The present disclosure will be described below based on examples. However, the above description and the following examples are provided for illustrative purposes only and are not intended to limit the present disclosure. Therefore, the scope of the present invention is not limited to the embodiments or examples specifically described herein, but is limited only by the claims.

[0290] The present disclosure will be explained in more detail below with reference to Reference Examples, Examples and Test Examples, but the present invention is not limited thereto.

[0291] The following abbreviations may be used in this specification: Ts: p-toluenesulfonyl THF: tetrahydrofuran TFA: trifluoroacetic acid DMF: N,N-dimethylformamide DMSO: dimethyl sulfoxide MeCN: acetonitrile Me: methyl Boc: tert-butoxycarbonyl Dess-Martin reagent: Dess-Martin periodinane (1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3-(1H)-one)

[0292] The NMR (Nuclear Magnetic Resonance) data used to identify the compounds was obtained using a JNM-ECS400 nuclear magnetic resonance spectrometer (400 MHz) manufactured by JEOL Ltd. The symbols used in NMR are: s, singlet; d, doublet; dd, double of doublets; t, triplet; td, double of triplets; q, quartet; m, multiplet; br, broad; brs, broad singlet; brm, broad multiplet; and J, coupling constant.

[0293] The LC / MS (Liquid Chromatography-Mass Spectrometry) analysis conditions used for compound identification are as follows: Among the observed mass spectrometry values ​​[MS (m / z)], the value corresponding to the monoisotopic mass (accurate mass consisting of only the main isotope) was defined as [M+H] ＋ , [M-H] － or [M+2H] ２＋ etc., and retention time is indicated as Rt (min).

[0294] Measurement Condition A Detector: ACQUITY (registered trademark) SQ detector (Waters) HPLC: ACQUITY UPLC (registered trademark) system Column: Waters ACQUITY UPLC (registered trademark) BEH C18 (1.7 μm, 2.1 mm × 30 mm) Solvent: Solution A: 0.06% formic acid / H ２ O, B solution: 0.06% formic acid / MeCN Gradient condition: 0.0-1.3min Linear gradient from B 2% to 96% Flow rate: 0.8mL / min UV: 220nm and 254nm Column temperature: 40℃

[0295] Measurement Condition B Detector: APCI 6120 Quadruple LC / MS (Agilent Technologies) HPLC: Agilent Technologies 1260 Infinity (registered trademark) system Column: Agilent Technologies (registered trademark) ZORBAX SB-C18 (1.8 μm, 2.1 mm × 50 mm) Solvent: Solution A: 0.1% formic acid / H ２ O, B solution: MeCN Gradient condition: 0.0-5.0min Linear gradient from B 5% to 90% Flow rate: 0.6mL / min UV: 210nm, 254nm, and 280nm Column temperature: 40℃

[0296] Measurement condition C Detection equipment: Shimadzu LCMS-2020 Column: L-column-2 ODS (4.6 mm x 35 mm) Gradient condition: MeCN / H ２ O / HCO ２ H=10 / 90 / 0.1 → 100 / 0 / 0.1 (0-2min), 100 / 0 / 0.1 (2-4min) Flow rate: 2mL / min Column temperature: 40℃

[0297] Reference Example 1: tert-Butyl 3-{[(4-acetyl-5-methoxypyridin-3-yl)oxy]methyl}-3-fluoroazetidine-1-carboxylate

[0298] Preparation of tert-butyl 3-{[(4-acetyl-5-methoxypyridin-3-yl)oxy]methyl}-3-fluoroazetidine-1-carboxylate (Reference Example 1) To a solution of 1-(3-hydroxy-5-methoxy-4-pyridinyl)ethanone (1.00 g) in THF (50.0 mL), tert-butyl 3-fluoro-3-(hydroxymethyl)azetidine-1-carboxylate (1.85 g) and triphenylphosphine (3.15 g) were added at room temperature, and the mixture was cooled to 0°C. Diisopropyl azodicarboxylate (2.5 mL) was added, and the mixture was stirred at 0°C for 12 hours. Saturated brine was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain Reference Example 1 (720 mg). LC-MS; [M+H] ＋ 355.2 / Rt (min) 0.886 (measurement condition A)

[0299] Reference Example 2: tert-Butyl 3-{[(3-acetyl-2-methoxypyridin-4-yl)oxy]methyl}-3-(hydroxymethyl)azetidine-1-carboxylate

[0300] a) Preparation of tert-butyl 3-(hydroxymethyl)-3-{[(4-methylbenzene-1-sulfonyl)oxy]methyl}azetidine-1-carboxylate Pyridine (5.60 mL), trimethylamine hydrochloride (0.13 g), and para-toluenesulfonyl chloride (1.45 g) were added to a solution of tert-butyl 3,3-bis(hydroxymethyl)azetidine-1-carboxylate (1.50 g) in methylene chloride (20.0 mL) at 0° C., and the mixture was stirred at room temperature for 12 hours. Saturated brine was added to the reaction mixture to quench the mixture, followed by extraction with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure to give the title compound (2.30 g). The resulting residue was used in the next reaction without purification. LC-MS [M+H] ＋ 372.1 / Rt (min) 0.928 (measurement condition A)

[0301] b) Preparation of tert-butyl 3-{[(3-acetyl-2-methoxypyridin-4-yl)oxy]methyl}-3-(hydroxymethyl)azetidine-1-carboxylate (Reference Example 2) 1-(4-hydroxy-2-methoxy-3-pyridinyl)ethanone (1.00 g) and cesium carbonate (7.80 g) were added to a solution of tert-butyl 3-(hydroxymethyl)-3-{[(4-methylbenzene-1-sulfonyl)oxy]methyl}azetidine-1-carboxylate (2.30 g) in DMF (10.0 mL), and the mixture was stirred at room temperature for 12 hours. Saturated brine was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and then filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain Reference Example 2 (1.01 g). LC-MS; [M+H] ＋ 367.3 / Rt (min) 0.796 (measurement condition A)

[0302] Reference Example 3: tert-butyl (3-{[3-(3-amino-1H-pyrazol-5-yl)-4-methoxypyridin-2-yl]oxy}propyl)carbamate

[0303] a) Preparation of tert-butyl {3-[(3-acetyl-4-methoxypyridin-2-yl)oxy]propyl}carbamate Cesium carbonate (7.80 g) and 3-(Boc-amino)propyl bromide (4.27 g) were added to a solution of 1-(2-hydroxy-4-methoxypyridin-3-yl)ethanone (2.00 g) in DMF (40.0 mL) at 0° C., and the mixture was stirred at room temperature for 12 hours. Saturated brine was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the title compound (2.25 g). LC-MS; [M+H] ＋ 325.2 / Rt (min) 0.959 (measurement conditions A) 1H-NMR (CDCl3) δ: 8.06 (1H, d, J = 6.0 Hz), 6.55 (1H, d, J = 6.0 Hz), 4.97 (1H, brs), 4.40 (2H, t, J = 6.4 Hz), 3.86 (3H, s), 3.27-3.24 (2H, m), 2.49 (3H, s), 1.96-1.90 (2H, m), 1.44 (9H, s).

[0304] b) Preparation of tert-butyl [3-({3-[(2E)-3-(dimethylamino)propyl-2-enyl]-4-methoxypyridin-2-yl}oxy)propyl]carbamate N,N-Dimethylformamide dimethyl acetal (10.0 mL) was added to a solution of tert-butyl {3-[(3-acetyl-4-methoxypyridin-2-yl)oxy]propyl}carbamate (2.25 g) in DMF (10.0 mL), and the mixture was stirred at 115° C. for 24 hours. After cooling, the solvent of the reaction solution was evaporated under reduced pressure to obtain the title compound (4.30 g) as a crude product. LC-MS; [M+H] ＋ 380.3 / Rt (min) 0.722 (measurement condition A)

[0305] c) Preparation of tert-butyl (3-{[4-methoxy-3-(1,2-oxazol-5-yl)pyridin-2-yl]oxy}propyl)carbamate Hydroxyamine hydrochloride (5.49 g) was added to a solution of tert-butyl [3-({3-[(2E)-3-(dimethylamino)propyl-2-enyl]-4-methoxypyridin-2-yl}oxy)propyl]carbamate (4.30 g) in ethanol (30.0 mL), and the mixture was stirred at 65° C. for 2 hours. After cooling, the reaction solution was added to a saturated aqueous solution of sodium hydrogen carbonate to quench the reaction. The resulting aqueous solution was extracted twice with ethyl acetate. The resulting organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, and then filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the title compound (1.63 g). LC-MS; [M+H] ＋ 350.2 / Rt (min) 0.722 (measurement conditions A) 1H-NMR (CDCl3) δ: 8.32 (1H, d, J = 1.6 Hz), 8.12 (1H, d, J = 6.0 Hz), 6.64 (1H, d, J = 6.0 Hz), 6.59 (1H, d, J = 1.6 Hz), 4.87 (1H, brs), 4.45 (2H, t, J = 6.4 Hz), 3.92 (3H, s), 3.28-3.24 (2H, m), 1.99-1.93 (2H, m), 1.44 (9H, s).

[0306] d) Preparation of tert-butyl (3-{[3-(cyanoacetyl)-4-methoxypyridin-2-yl]oxy}propyl)carbamate Potassium hydroxide (0.29 g) was added to a mixed solution of tert-butyl (3-{[4-methoxy-3-(1,2-oxazol-5-yl)pyridin-2-yl]oxy}propyl)carbamate (1.63 g) in ethanol (20.0 mL) and water (5.00 mL), and the mixture was stirred at room temperature for 5 hours. The reaction solution was concentrated under reduced pressure, and the residue was added to saturated brine for quenching. The resulting aqueous solution was extracted twice with ethyl acetate. The resulting organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, then filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the title compound (1.50 g). LC-MS; [M+H] ＋ 350.2 / Rt (min) 0.872 (measurement conditions A) 1H-NMR (CDCl3) δ: 8.14 (1H, d, J = 6.0 Hz), 6.59 (1H, d, J = 6.0 Hz), 4.94 (1H, brs), 4.44 (2H, t, J = 6.0 Hz), 3.92 (3H, s), 3.29-3.23 (2H, m), 1.99-1.93 (2H, m), 1.43 (9H, s).

[0307] e) Preparation of tert-butyl (3-{[3-(3-amino-1H-pyrazol-5-yl)-4-methoxypyridin-2-yl]oxy}propyl)carbamate (Reference Example 3) Acetic acid (3.28 mL) and hydrazine monohydrate (2.78 mL) were added to a solution of tert-butyl (3-{[3-(cyanoacetyl)-4-methoxypyridin-2-yl]oxy}propyl)carbamate (1.99 g) in ethanol (20.0 mL) at 0° C., and the mixture was stirred at 90° C. for 24 hours. After cooling, the reaction solution was quenched by adding it to a saturated aqueous solution of sodium bicarbonate. The resulting aqueous solution was extracted twice with chloroform. The resulting organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, then filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform / methanol) to obtain Reference Example 3 (1.60 g). LC-MS; [M+H]＋ 364.0 / Rt (min) 0.684 (measurement conditions A) 1H-NMR (DMSO-D6) δ: 11.61-10.91 (1H, brs), 8.01 (1H, d, J = 12.0 Hz), 6.93-6.89 (1H, m), 6.84 (1H, d, J = 6.0 Hz), 5.93 (1H, brs), 4.50 (1H, brs), 4.32 (2H, t, J = 6.4 Hz), 3.89 (3H, s), 3.09-3.04 (2H, m), 1.86-1.79 (2H, m), 1.37 (9H, s).

[0308] Reference Example 4: tert-Butyl (3-{[3-(3-amino-1H-pyrazol-5-yl)-6-chloro-4-methoxypyridin-2-yl]oxy}propyl)carbamate

[0309] a) Preparation of 1-{6-chloro-4-methoxy-2-[(4-methoxyphenyl)methoxy]pyridin-3-yl}ethan-1-ol A solution of 2-chloro-4-methoxy-6-[(4-methoxybenzyl)oxy]pyridine (10.0 g) in THF (100 mL) was cooled to −78°C. 2.8 mol / L n-butyllithium (15.3 mL) was added, and the mixture was stirred at −78°C for 3 hours. Acetamide (6.30 mL) was added, and the mixture was stirred at −78°C for 6 hours. The reaction mixture was quenched by adding saturated aqueous sodium bicarbonate solution, and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and then filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the title compound (4.25 g). LC-MS; [M+H] ＋324.2 / Rt (min) 1.118 (measurement conditions A) 1H-NMR (CDCl3) δ: 7.36 (2H, d, J = 8.8 Hz), 6.89 (2H, d, J = 8.8 Hz), 6.55 (1H, s), 5.32 (2H, s), 5.16-5.11 (1H, m), 3.85 (3H, s), 3.80 (3H, s), 3.30 (1H, s), 1.41 (3H, d, J = 6.8 Hz). 13C-NMR (CDCl3) δ: 165.2, 159.6, 147.6, 130.1, 128.6, 114.1, 112.6, 101.8, 68.7, 62.9, 56.3, 55.4, 23.1.

[0310] b) Preparation of 1-{6-chloro-4-methoxy-2-[(4-methoxyphenyl)methoxy]pyridin-3-yl}ethan-1-one. Dess-Martin reagent (6.19 g) was added to a solution of 1-{6-chloro-4-methoxy-2-[(4-methoxyphenyl)methoxy]pyridin-3-yl}ethan-1-ol (3.15 g) in methylene chloride (100 mL), and the mixture was stirred at room temperature for 12 hours. The reaction solution was quenched by adding it to a saturated aqueous solution of sodium bicarbonate. The resulting aqueous solution was extracted twice with ethyl acetate. The resulting organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, then filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the title compound (2.50 g). LC-MS; [M+H] ＋ 322.2 / Rt (min) 1.081 (measurement condition A)

[0311] c) Preparation of 1-(6-chloro-2-hydroxy-4-methoxypyridin-3-yl)ethan-1-one TFA (6.00 mL) was added to a solution of 1-{6-chloro-4-methoxy-2-[(4-methoxyphenyl)methoxy]pyridin-3-yl}ethan-1-one (2.50 g) in methylene chloride (30.0 mL), and the mixture was stirred at room temperature for 2 hours. The solvent was evaporated under reduced pressure, and the residue was purified by amine silica gel column chromatography (chloroform / methanol) to obtain the title compound (2.00 g). LC-MS; [M+H] ＋202.1 / Rt (min) 0.773 (measurement condition A)

[0312] d) Preparation of tert-butyl {3-[(3-acetyl-6-chloro-4-methoxypyridin-2-yl)oxy]propyl}carbamate Cesium carbonate (9.70 g) and 3-(Boc-amino)propyl bromide (5.63 g) were added to a solution of 1-(6-chloro-2-hydroxy-4-methoxypyridin-3-yl)ethan-1-one (3.00 g) in DMF (50.0 mL) at 0° C., and the mixture was stirred at room temperature for 12 hours. Saturated brine was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the title compound (1.01 g). LC-MS; [M+H] ＋ 359.3 / Rt (min) 1.123 (measurement condition A)

[0313] e) Preparation of tert-butyl [3-({6-chloro-3-[(2E)-3-(dimethylamino)propyl-2-enyl]-4-methoxypyridin-2-yl}oxy)propyl]carbamate N,N-Dimethylformamide dimethyl acetal (3.00 mL) was added to a solution of tert-butyl {3-[(3-acetyl-6-chloro-4-methoxypyridin-2-yl)oxy]propyl}carbamate (840 mg) in DMF (10.0 mL), and the mixture was stirred at 115° C. for 24 hours. After cooling, the solvent of the reaction solution was evaporated under reduced pressure to obtain the title compound (0.97 g) as a crude product. LC-MS; [M+H] ＋ 414.4 / Rt (min) 0.944 (measurement condition A)

[0314] f) Preparation of tert-butyl (3-{[6-chloro-4-methoxy-3-(1,2-oxazol-5-yl)pyridin-2-yl]oxy}propyl)carbamate Hydroxyamine hydrochloride (1.70 g) was added to a solution of tert-butyl [3-({6-chloro-3-[(2E)-3-(dimethylamino)propyl-2-enyl]-4-methoxypyridin-2-yl}oxy)propyl]carbamate (0.97 g) in ethanol (30.0 mL), and the mixture was stirred at 65° C. for 2 hours. After cooling, the reaction solution was added to a saturated aqueous solution of sodium hydrogen carbonate to quench the reaction. The resulting aqueous solution was extracted twice with ethyl acetate. The resulting organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, and then filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the title compound (677 mg). LC-MS; [M+H] ＋ 384.3 / Rt (min) 1.132 (measurement condition A)

[0315] g) Preparation of tert-butyl (3-{[6-chloro-3-(cyanoacetyl)-4-methoxypyridin-2-yl]oxy}propyl)carbamate Potassium hydroxide (100 mg) was added to a mixed solution of tert-butyl (3-{[6-chloro-4-methoxy-3-(1,2-oxazol-5-yl)pyridin-2-yl]oxy}propyl)carbamate (677 mg) in ethanol (20.0 mL) and water (5.00 mL), and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was added to saturated brine for quenching. The resulting aqueous solution was extracted twice with ethyl acetate. The resulting organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, then filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the title compound (470 mg). LC-MS; [M+H] ＋ 384.3 / Rt (min) 1.064 (measurement condition A)

[0316] h) Preparation of tert-butyl (3-{[3-(3-amino-1H-pyrazol-5-yl)-6-chloro-4-methoxypyridin-2-yl]oxy}propyl)carbamate (Reference Example 4) Acetic acid (0.71 mL) and hydrazine monohydrate (0.77 mL) were added to a solution of tert-butyl (3-{[6-chloro-3-(cyanoacetyl)-4-methoxypyridin-2-yl]oxy}propyl)carbamate (470 mg) in ethanol (20.0 mL) at 0° C., and the mixture was stirred at 90° C. for 24 hours. After cooling, the reaction solution was quenched by adding it to a saturated aqueous solution of sodium hydrogen carbonate. The resulting aqueous solution was extracted twice with chloroform. The resulting organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, then filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform / methanol) to obtain Reference Example 4 (350 mg). LC-MS; [M+H] ＋ 398.3 / Rt (min) 0.911 (measurement condition A)

[0317] Reference Example 5: 1-(4-hydroxy-6-methoxypyrimidin-5-yl)ethan-1-one

[0318] a) Preparation of 1-(4-hydroxy-6-methoxypyrimidin-5-yl)ethan-1-one To a solution of 1-(4,6-dimethoxypyrimidin-5-yl)ethan-1-one (2.00 g) in dichloromethane (30.0 mL) was added a solution of boron tribromide in dichloromethane (1.0 mol / L, 54.9 mL) at −60° C., and the mixture was stirred at −50° C. or below for 3 hours. The reaction mixture was quenched by adding saturated saline solution, and then extracted with chloroform. The organic layer was washed with saturated saline solution and dried over anhydrous sodium sulfate, and then filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (methanol / chloroform) to obtain the title compound (220 mg). LC-MS; [M+H] ＋ 169.1 / Rt (min) 0.367 (measurement conditions A) 1H-NMR (CDCl3) δ: 14.31 (1H, brs), 8.36 (1H, s), 4.10 (3H, s), 2.65 (3H, s).

[0319] Reference Example 6: tert-butyl (3-{[4-acetyl-5-(fluoromethoxy)pyridin-3-yl]oxy}propyl)carbamate

[0320] a) Preparation of 3-fluoro-5-(fluoromethoxy)pyridine Cesium carbonate (4.32 g) and fluoromethyl 4-methylbenzenesulfonate (2.17 g) were added to a solution of 5-fluoropyridin-3-ol (1.00 g) in DMF (30.0 mL) at room temperature, and the mixture was heated and stirred at 80°C for 8 hours. Water was added to the reaction mixture, and the mixture was extracted with diethyl ether. The organic layer was dried over anhydrous sodium sulfate, filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the title compound (538 mg). LC-MS; [M+H] ＋ 146.0 / Rt (min) 0.574 (measurement condition A)

[0321] b) Preparation of 1-[3-fluoro-5-(fluoromethoxy)pyridin-4-yl]ethan-1-ol A solution of N,N-diisopropylethylamine (0.687 mL) in THF (15 mL) was cooled to -78°C, n-butyllithium (1.58 mol / L, 3.05 mL) was added, and the mixture was stirred at 0°C for 15 minutes. The reaction mixture was cooled again to -78°C, 3-fluoro-5-(fluoromethoxy)pyridine (538 mg) was added, and the mixture was stirred for 1 hour, followed by the addition of acetaldehyde (0.419 mL). The reaction mixture was gradually warmed to room temperature and stirred overnight. Water was then added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and then filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the title compound (475 mg). LC-MS; [M+H] ＋190.1 / Rt (min) 0.494 (measurement conditions A) 1H-NMR (CDCl3) δ:8.30 (1H, d, J = 1.8 Hz), 8.26 (1H, s), 5.85 (1H, dd, J = 6.7, 3.1 Hz), 5.71(1H, dd, J = 7.2, 3.2 Hz), 5.31-5.22 (1H, m), 2.60 (1H, dd, J = 9.8, 1.8 Hz),1.59 (3H, d, J = 6.7 Hz).

[0322] c) Preparation of 1-[3-fluoro-5-(fluoromethoxy)pyridin-4-yl]ethan-1-one A solution of 1-[3-fluoro-5-(fluoromethoxy)pyridin-4-yl]ethan-1-ol (475 mg) in dichloromethane (10.0 mL) was ice-cooled, Dess-Martin reagent (1.60 g) was added, and the mixture was stirred at room temperature overnight. Aqueous sodium thiosulfate solution and saturated aqueous sodium bicarbonate solution were added to the reaction mixture, and the mixture was extracted with chloroform. The resulting organic layer was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the title compound (407 mg). LC-MS; [M+H] ＋ 188.0 / Rt (min) 0.618 (measurement condition A)

[0323] d) Preparation of tert-butyl (3-{[4-acetyl-5-(fluoromethoxy)pyridin-3-yl]oxy}propyl)carbamate To a solution of 1-[3-fluoro-5-(fluoromethoxy)pyridin-4-yl]ethan-1-one (407 mg) and cesium carbonate (1.42 g) in DMF (10.0 mL), tert-butyl (3-hydroxypropyl)carbamate (762 mg) was added, and the mixture was heated and stirred at 80°C for 8 hours. Water was added to the reaction solution, and the mixture was extracted with diethyl ether. The organic layer was dried over anhydrous sodium sulfate, filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the title compound (67 mg). LC-MS; [M+H] ＋343.2 / Rt (min) 0.832 (measurement conditions A) 1H-NMR(CDCl3) δ:8.22 (1H, d, J = 1.8 Hz), 8.16 (1H, s), 5.68 (2H, d, J = 53.6 Hz), 4.70 (1H, brs), 4.15 (2H, t, J = 6.1 Hz), 3.32-3.21 (2H, m), 2.50 (3H, s), 2.00-1.93 (2H,m), 1.41 (9H, s).

[0324] Reference Example 7: tert-butyl (3-{[4-methoxy-3-(1,2-oxazol-5-yl)pyridin-2-yl]oxy}propyl)carbamate

[0325] a) Preparation of (2E)-3-(dimethylamino)-1-(2-hydroxy-4-methoxypyridin-3-yl)prop-2-en-1-one To a mixture of 1-(2-hydroxy-4-methoxypyridin-3-yl)ethanone (2.50 g) and toluene (6.23 ml), tert-butoxybis(dimethylamino)methane (5.21 g) was added, and the mixture was heated and stirred at 55°C for 6 hours. After cooling, toluene (18.6 ml) was added dropwise, and the resulting crystals were collected by filtration and washed with toluene. The crystals were dried in vacuo to obtain the title compound (2.87 g). The filtrate was concentrated under reduced pressure, and the crystals precipitated from an ethanol / toluene mixed solvent were washed with an ethanol / toluene mixed solvent and dried in vacuo to obtain the second crystal of the title compound (0.20 g). LC-MS; [M+H] ＋ 223.1 / Rt (min) 1.564 1H-NMR (DMSO-D6) δ: 11.25 (1H, br s), 7.39 (1H, d, J = 6.7 Hz), 7.11 (1H, br s), 6.21(1H, d, J = 7.9 Hz), 5.00 (1H, d, J = 11.0 Hz), 3.73 (3H, s), 3.00 (3H, br s),2.75 (3H, br s).

[0326] b) Preparation of 4-methoxy-3-(1,2-oxazol-5-yl)pyridin-2-ol Hydroxyamine hydrochloride (1.56 g) was added to a mixture of (2E)-3-(dimethylamino)-1-(2-hydroxy-4-methoxypyridin-3-yl)prop-2-en-1-one (2.50 g) and ethanol (37.5 ml), and the mixture was stirred at room temperature for 16 hours. Water (37.5 ml) was added to the reaction mixture, and the mixture was concentrated under reduced pressure to 28.5 g. Further water (9.0 ml) was added, and the mixture was stirred at 0°C. The resulting crystals were collected by filtration, washed with cold water, and dried in vacuo to give the title compound (1.53 g). LC-MS [M+H] ＋ 193.1 / Rt (min) 0.928 1H-NMR (DMSO-D6) δ: 11.83 (1H, br s), 8.51 (1H, d, J =1.8 Hz), 7.61 (1H, d, J = 7.3 Hz), 6.83 (1H, d, J = 1.8 Hz), 6.38 (1H, d, J =7.3 Hz), 3.93 (3H, s).

[0327] c) Preparation of 4-methoxy-3-(1,2-oxazol-5-yl)pyridin-2-yl trifluoromethanesulfonate Trifluoromethanesulfonic anhydride (2.16 g) was added dropwise to a mixture of 4-methoxy-3-(1,2-oxazol-5-yl)pyridin-2-ol (1.23 g) and pyridine (6.1 ml) at 0°C. The reaction mixture was warmed and stirred at room temperature for 16 hours. The reaction mixture was cooled to 0°C, water (16 ml) was added dropwise, and the mixture was stirred at 0°C for 1.5 hours. The resulting crystals were collected by filtration, washed with a water / pyridine mixed solvent and then with water, and dried in vacuo to give the title compound (1.91 g). LC-MS; [M+H] ＋ 325.0 / Rt (min) 4.781 1H-NMR (CDCl3) δ: 8.37 (1H, d, J = 1.8 Hz), 8.32(1H, d, J = 6.1 Hz), 7.01 (1H, d, J = 6.1 Hz), 6,67 (1H, d, J = 1.8 Hz), 4.00(3H, s).

[0328] d) Preparation of tert-butyl (3-{[4-methoxy-3-(1,2-oxazol-5-yl)pyridin-2-yl]oxy}propyl)carbamate Diisopropylethylamine (72 mg) was added to a mixture of 4-methoxy-3-(1,2-oxazol-5-yl)pyridin-2-yl trifluoromethanesulfonate (50 mg), tert-butyl (3-hydroxypropyl)carbamate (54 mg), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (18 mg), tris(dibenzylideneacetone)dipalladium(0) chloroform adduct (16 mg), and toluene (1.5 ml), and the mixture was heated and stirred at 100°C for 4 hours. After allowing the reaction mixture to cool, insoluble matter was removed by filtration through Celite and washed with ethyl acetate. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the title compound (30 mg). LC-MS; [M+H] ＋ 350.2 / Rt (minutes) 4.647

[0329] Reference Examples 8 to 41: Using the corresponding starting compounds, the compounds of Reference Examples 8 to 41 shown in the table below were obtained in the same manner as in Reference Examples 1 to 7.

[0330] Reference Example 42: tert-butyl {3-[(3-{3-[(5-cyanopyrazin-2-yl)amino]-1H-pyrazol-5-yl}-4-methoxypyridin-2-yl)oxy]propyl}carbamate

[0331] To a solution of tert-butyl (3-{[3-(3-amino-1H-pyrazol-5-yl)-4-methoxypyridin-2-yl]oxy}propyl)carbamate (600 mg) in DMSO (10.0 mL) were added 4-ethylmorpholine (1.00 mL) and 5-chloropyrazine-2-carbonitrile (461 mg) at room temperature, and the mixture was stirred at 80°C for 12 hours. Saturated brine was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to give the title compound (1.01 g). LC-MS; [M+H] ＋ 467.3 / Rt (min) 0.885 (measurement conditions A) 1H-NMR (DMSO-D6) δ: 12.4 (1H, s), 10.7 (1H, s), 8.61 (1H, s), 8.50 (1H, brs), 8.07 (1H, d, J = 5.2 Hz), 7.06 (1H, brs), 6.90 (1H, d, J = 6.0 Hz), 4.34 (2H, t, J = 6.4 Hz), 3.93 (3H, s), 3.14-3.08 (2H, m), 1.87-1.84 (2H, m), 1.35 (9H, s).

[0332] Reference Examples 43 to 78: Using the corresponding starting compounds, the compounds of Reference Examples 43 to 78 shown in the table below were obtained in the same manner as in Reference Example 42.

[0333] Reference Example 79: tert-butyl {3-[(3-{3-[(5-chloropyrazin-2-yl)amino]-1H-pyrazol-5-yl}-4-methoxypyridin-2-yl)oxy]propyl}carbamate

[0334] To a solution of tert-butyl (3-{[3-(3-amino-1H-pyrazol-5-yl)-4-methoxypyridin-2-yl]oxy}propyl)carbamate (60 mg) in 1,4-dioxane (825 μL), 2,5-dichloropyrazine (32.9 μL), tris(dibenzylideneacetone)dipalladium(0) (15.1 mg), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (19.1 mg), and cesium carbonate (108 mg) were added at room temperature, and the mixture was stirred at 150° C. for 2 hours under microwave irradiation. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, then filtered off, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the title compound. LC-MS; [M+H] ＋ 476.3 / Rt (min) 0.969 (measurement condition A)

[0335] Example 1: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile

[0336] To a solution of tert-butyl {3-[(3-{3-[(5-cyanopyrazin-2-yl)amino]-1H-pyrazol-5-yl}-4-methoxypyridin-2-yl)oxy]propyl}carbamate (200 mg) obtained in Reference Example 42 in methylene chloride (20.0 mL) was added TFA (2.00 mL) at room temperature, and the mixture was stirred at room temperature for 1 hour. The solvent was evaporated under reduced pressure, and the residue was purified by amine silica gel column chromatography (ethyl acetate / methanol) to obtain Example 1 (112 mg). LC-MS; [M+H] ＋367.2 / Rt (min) 0.671 (measurement conditions A) 1H-NMR (DMSO-D6) δ: 8.65 (1H, s), 8.50 (1H, brs), 8.06 (1H, d, J = 6.4 Hz), 7.04 (1H, brs), 6.89 (1H, d, J = 6.4 Hz), 4.41 (2H, t, J = 6.0 Hz), 3.92 (3H, s), 2.75 (1H, t, J = 6.8 Hz), 1.85-1.76 (2H, m).

[0337] Examples 2 to 38: Using the corresponding starting compounds, the compounds of Examples 2 to 38 shown in the table below were obtained in the same manner as in Example 1.

[0338] Powder X-ray diffraction measurements of Examples 39 to 44 were performed under the conditions shown below. The obtained diffraction patterns (XRD spectra) are shown in Figures 1 to 6. Examples 39 to 41 are crystals of various salts of the compound of Example 1, and Examples 42 to 44 are crystalline polymorphs of the compound of Example 1.

[0339] The crystal form can be identified based on the characteristic diffraction peaks of each crystal shown in the diffraction patterns of FIGS.

[0340] The main diffraction peaks and characteristic diffraction peaks identified from the diffraction patterns of Figures 1 to 6 are listed below. Note that the diffraction peak values ​​at the diffraction angle 2θ (°) described in the following examples may be subject to some measurement error depending on the measuring device or measurement conditions. Specifically, the measurement error may be within a range of ±0.2, preferably ±0.1.

[0341] Powder X-ray diffraction measurement method: Detector: Spectris Power X-ray diffraction system Empyrian X-ray tube: CuKα (wavelength: 1.54 angstroms) Tube voltage: 45 kV Tube current: 40 mA Measurement range: 4 to 40 degrees (2θ) Step width: 0.013 degrees Integration time: 100 seconds / step

[0342] Example 39: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile hydrochloride

[0343] Methanol (0.6 mL) was added to the compound of Example 1 (30.0 mg), and then 5-10% (W / V) hydrochloric acid-methanol solution (60 μL) was added, followed by stirring at a set temperature of 60°C for 2 hours. After cooling, the mixture was left to stand overnight, and the precipitated solid was collected by filtration and dried to obtain the title compound as a crystal (Form I). [Form I] The powder X-ray diffraction pattern is shown in Figure 1. Main diffraction peaks: 2θ (°) = 7.2, 8.8, 9.8, 10.2, 10.7, 16.7, 18.5, 26.2, 27.0 Characteristic diffraction peaks: 2θ (°) = 7.2, 8.8, 9.8, 10.2, 10.7

[0344] Example 40: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile phosphate

[0345] To the compound of Example 1 (30.0 mg), a 13.4 mg / mL phosphoric acid-methanol solution (1200 μL) was added, and the mixture was stirred at a set temperature of 60°C for 4 hours. After cooling, the mixture was left standing overnight, and the precipitated solid was collected by filtration and dried to obtain the title compound as a crystal (Form II). The powder X-ray diffraction pattern for [Form II] is shown in Figure 2. Main diffraction peaks: 2θ (°) = 6.8, 7.5, 11.7, 11.9, 13.0, 16.4, 19.3, 20.4, 22.7, 24.3 Characteristic diffraction peaks: 2θ (°) = 6.8, 7.5, 11.7, 11.9, 13.0

[0346] Example 41: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile tosylate

[0347] Methanol (0.6 mL) was added to the compound of Example 1 (30.0 mg), and then a 26.0 mg / mL tosylic acid-methanol solution (0.6 mL) was added, followed by stirring at a set temperature of 60°C for 2 hours. After cooling, the mixture was left standing overnight, and the precipitated solid was collected by filtration and dried to obtain the title compound as a crystal (Form III). [Form III] The powder X-ray diffraction pattern is shown in Figure 3. Main diffraction peaks: 2θ (°) = 6.0, 9.0, 12.1, 14.4, 16.2, 17.0, 22.8, 26.3 Characteristic diffraction peaks: 2θ (°) = 6.0, 9.0, 12.1, 14.4, 16.2, 17.0

[0348] Example 42: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile

[0349] [Form IV] The powder X-ray diffraction pattern is shown in Figure 4. The title compound was obtained as a crystal (Form IV) according to Example 1. Main diffraction peaks: 2θ (°) = 9.3, 10.2, 10.7, 13.6, 16.7, 17.1, 17.8, 18.6, 26.1, 26.4 Characteristic diffraction peaks: 2θ (°) = 9.3, 10.2, 10.7, 16.7, 26.1, 26.4

[0350] Example 43: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile

[0351] [Form V] The powder X-ray diffraction pattern is shown in Figure 5. A solution of the compound of Example 1 (5 mg) in tetrahydrofuran / water (10 / 1, 0.5 mL) was added and heated at a set temperature of 105°C for 1 hour. After cooling, the container was sealed and left to stand for 4 days, then opened and left to stand for 3 days, and the solvent was distilled off. The precipitated solid was obtained as the title compound as a crystal (Form V). Main diffraction peaks: 2θ (°) = 7.9, 8.7, 12.2, 13.1, 15.9, 17.6, 19.9, 21.9, 22.8, 26.6 Characteristic diffraction peaks: 2θ (°) = 7.9, 8.7, 12.2, 13.1, 15.9, 26.6

[0352] Example 44: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridin-3-yl]-1H-pyrazol-3-yl}amino)pyrazine-2-carbonitrile

[0353] [Form VI] The powder X-ray diffraction pattern is shown in Figure 6. A solution of the compound of Example 1 (5 mg) in acetone / water (10 / 1, 0.5 mL) was added and heated at a set temperature of 105°C for 1 hour. After cooling, the container was sealed and left to stand for 4 days, then opened and left to stand for 3 days, and the solvent was distilled off. The precipitated solid was obtained as the title compound as a crystal (Form VI). Main diffraction peaks: 2θ (°) = 5.3, 5.7, 7.0, 7.3, 7.8, 8.4, 9.3, 10.5, 11.5, 14.1 Characteristic diffraction peaks: 2θ (°) = 5.3, 5.7, 7.0, 7.3, 8.4, 9.3

[0354] Test Example Test Example 1: CHK1 inhibitory activity test IMAP TR-FRET Screening Express Kit (R8160) was obtained from Molecular Device, Inc. CHK1 kinase (02-117, Carna Bio), FAM-labeled CHK1tide (R7185, Molecular Device, Inc.), and ATP were diluted with assay buffer to final concentrations of 4 μg / mL, 2 μM, and 20 μM, respectively. FAM-PKAtide (R7255, Molecular Device) and FAM-Phospho-PKAtide (R7304, Molecular Device) were diluted and mixed to create a calibration standard series with phosphorylation levels ranging from 0 to 100%. 5 μL of compound dissolved in 0.4% DMSO was added to a 384-well plate. A compound test group was added with 5 μL each of CHK1, CHK1tide, and ATP, and a standard group was added with 20 μL each of the standard. The kinase reaction was carried out at 30°C for 3 hours. Then, 60 μL of Binding Reagent (80% Buffer A, 20% Buffer B, 1:600 ​​Binding Reagent, 1:400 Tb-Donor) was added, and the binding reaction was carried out at room temperature for 2 hours. Fluorescence intensities at 520 nm and 490 nm upon excitation at 340 nm were measured using a SpectraMax Paradigm (Molecular Devices). The phosphorylation level of CHK1tide was calculated using the standard. The DMSO-treated group was set to a phosphorylation level of 100%, and kinase activity was calculated using the formula below. The IC40 corresponding to the concentration of the compound at which kinase activity was 50% was calculated. ５０ The value was calculated.

[0355] Kinase inhibition (%) = 100 - A / B x 100 A: Signal in the presence of the compound to be evaluated B: Signal in the negative control (DMSO-treated group)

[0356] The compounds obtained in the Examples and Plexasertib purchased from MedChemExpress (the same supplier was used in the following Test Examples) were subjected to the test shown in Test Example 1. ５０The values ​​(nM) are shown in the table below.

[0357] As shown in the table above, the compounds of Examples 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 17, 18, 19, 20, 21, 22, 24, 26, 28, 30, and 37 exhibited stronger inhibitory effects on CHK1 activity than prexasertib. Among them, the compounds of Examples 1 and 8 have particularly strong and exceptional effects of inhibiting CHK1 activity.

[0358] Test Example 2: Cell proliferation inhibition experiment ES-2 cells were obtained from the American Type Culture Collection (ATCC). The cells were cultured in McCoy's 5a medium containing 10% fetal bovine serum and 1% penicillin / streptomycin at 37°C and 5% CO ２ 500 cells were seeded per well in a 384-well plate, and the test compound was added so that the final DMSO concentration was 0.1%, followed by culturing for 2 days. After the culture was completed, the cell viability was calculated using CellTiter-Glo® 3D Reagent (Promega, G968B). From the viability curve, the IC value, which corresponds to the concentration of the test compound that shows 50% inhibition of cell proliferation, was determined. ５０ The values ​​were calculated. ES-2 cells are known to exhibit cisplatin resistance.

[0359] The compounds obtained in the Examples and Prexasertib were subjected to the test shown in Test Example 2. The concentration of each test substance that inhibits 50% of cell growth (IC ５０ The cell proliferation inhibition test results for representative compounds of the present disclosure are listed in the table below, with the rankings I, II, III, IV, and V (1000 nM<I, 1000 nM>II>300 nM, 300 nM>III>100 nM, 100 nM>IV>30 nM, and 30 nM>V).

[0360] As shown in the table above, the compounds of Examples 1, 3, 4, 5, 6, 11, 15, 17, 19, 20, 21, 22, 23, 25, 26, 27, 28, 29, 30, 31, 32, 33, 35, 36, 37 and 38 exhibited excellent cell proliferation inhibitory effects equivalent to those of prexasertib.

[0361] Test Example 3: hERG current inhibition test Test substances were added to cultured CHO cell lines stably expressing the hERG (human Ether-a-go-go Related Gene) gene at concentrations of 0.27 to 100 μM. hERG current under voltage stimulation was measured using Qube384 (Sophion Bioscience), and the concentration (IC) of each test substance at which the hERG current was inhibited by 50% was determined. ５０ The values ​​(μM) were calculated.

[0362] The compounds obtained in the Examples and Prexasertib were subjected to the test shown in Test Example 3. In addition, for the compounds of each Example, the IC value of hERG obtained in Test Example 3 was ５０ The IC value of CHK1 inhibition obtained in Test Example 1 ５０ The value obtained by dividing by was calculated as hERG / CHK1. The results are shown in the table below.

[0363] As shown in the table above, the compounds of Examples 1, 3, 4, 7 to 10, 14, 24, 32 and 34 had IC values ​​of CHK1 inhibition. ５０ and hERG IC ５０ Among them, the compounds of Examples 1, 3, 7, 8, 14 and 24 had an IC value of 50,000 times or more for CHK1 inhibition. ５０ and hERG IC ５０ These six compounds showed a difference of 18 times or more compared to prexasertib, and have heterogeneous effects with high safety.

[0364] Test Example 4: Evaluation of CYP3A4 MBI and Enzyme Inactivation Clearance Cytochrome P450 (hereinafter referred to as CYP) is the most important enzyme group involved in drug metabolism, and many pharmacokinetic interactions are based on the inhibition of these CYP activities. There are multiple molecular species of CYP, and CYP3A4 in particular is most involved in the metabolism of drugs in CYP-mediated oxidation reactions, and accounts for the majority of CYP present in the liver.

[0365] There are two main types of CYP inhibition: "reversible inhibition" and "irreversible inhibition (mechanism-based inhibition: MBI)." In particular, MBI-based CYP inhibition is known to have the potential to cause not only drug interactions but also serious side effects (such as hepatotoxicity) (Curr Opin Drug Discover Devel. 2010 January, 13(1), 66-77, Therapeutics and Clinical Risk Management, 2005, 1(1), 3-13).

[0366] The compound of this example and prexasertib were used to evaluate MBI and enzyme inactivation clearance of CYP3A4.

[0367] Using human liver microsomes as the enzyme source and midazolam or testosterone as the substrate, the inhibitory effect and mode of inhibition of the test compound against CYP3A4 were evaluated. After a metabolic reaction at 37°C for 30 minutes, the substrate-derived metabolic products in the test compound-added group (4 concentrations) and the substrate-derived metabolic products in the non-added group were measured by LC-MS / MS, and the inhibition rate was calculated from the ratio. In addition, the IC ５０ If the test compound exhibits MBI activity, the IC value was calculated by pre-incubating the compound in the presence of NADPH (cofactor) and then adding a substrate to initiate the reaction. ５０ Since it is known that pre-incubation reduces the IC ５０ If the value changed by 2-fold or more, it was judged that there was an MBI effect.

[0368] If MBI action is observed, k ｉｎａｃｔ(maximum enzyme inactivation rate constant) and K Ｉ The enzyme inactivation clearance (CL) (the concentration of the inhibitor that results in 50% of the maximum enzyme inactivation rate) was calculated by the nonlinear least squares method. Furthermore, the enzyme inactivation clearance (CL) was calculated according to the method described in Drug Metabolism and Disposition, 2011, 39(7), 1247-1254. ｉｎｔ ＝ k ｉｎａｃｔ / K Ｉ (ml / min / mmol)×CYP contents (pmol / mg protein)).

[0369] Pre-incubation IC as an index of MBI action on CYP3A4 of each example compound and prexasertib ５０ The fold change in values ​​and enzyme inactivation clearance are shown in the table below.

[0370] The present inventors have newly discovered that prexasertib inhibits CYP3A4 based on MBI, and therefore there is a risk of causing serious side effects such as hepatotoxicity.

[0371] As shown in the table above, it was revealed that although prexasertib exhibits MBI-based inhibition of CYP3A4, the compounds of Examples 1, 2, 4, 6 to 8, 14, 29 to 31, 34, 37, and 38 do not exhibit MBI-based inhibition of CYP3A4. These results indicate that the compounds of Examples 1, 2, 4, 6 to 8, 14, 29 to 31, 34, 37, and 38 have heterogeneous effects, such as high safety with reduced risk of not only drug interactions but also serious side effects such as hepatotoxicity. Among these, the compound represented by formula (3) exhibits particularly high safety and heterogeneous effects.

[0372] ​Test Example 5. Human bone marrow cell colony formation test (blood toxicity evaluation test) Human bone marrow CD34-positive hematopoietic stem cells were obtained from Lonza Co., Ltd. The frozen cells were thawed in a 9:1 mixture of Methocult Express medium and RPMI 1640 medium containing 10% fetal bovine serum and 1% penicillin / streptomycin, and 5,000 cells were seeded per well in a 6-well plate. The cells were incubated at 37°C and 5% CO ２ The cells were cultured overnight in the presence of DMSO. The compound to be evaluated was added to a final concentration of 0.1% and 100 nM DMSO, and the cells were cultured for 48 hours. The cells were harvested and washed with PBS. One-sixth of the amount of cells was seeded into a 12-well plate and incubated at 37°C, 5% CO ２ After completion of the culture, the number of colonies formed was counted using thiazolyl blue tetrazolium bromide (MTT).

[0373] The representative compounds obtained in the Examples and Prexasertib were subjected to the test shown in Test Example 5. The results are shown in the table below.

[0374] As shown in the above table, it was revealed that 100 nM of prexasertib exhibited an inhibitory effect on human bone marrow cell colony formation, whereas 100 nM of Example 1 did not exhibit an inhibitory effect on human bone marrow cell colony formation. From these results, the compound of Example 1 has a heterogeneous effect with high safety in terms of hematotoxicity.

[0375] Test Example 6 Liposome Encapsulation Test A liposome encapsulation test was carried out for representative Example compounds and Prexasertib.

[0376] 6.48 g of hydrogenated soybean phosphatidylcholine (COATSOME NC-21E, NOF Corporation), 2.32 g of cholesterol (Sigma), and 2.06 g of distearoylphosphatidylethanolamine-methoxypolyethylene glycol 2000 (SUNBRIGHT DSPE-020CN, NOF Corporation) were dissolved in 560 mL of t-butyl alcohol heated to 65°C. This solution was frozen in a dry ice-acetone bath, and then the t-butyl alcohol was removed by distillation under reduced pressure to obtain a lipid mixture.

[0377] To the lipid mixture, 200 mL of 250 mM ammonium sulfate solution was added, heated to 65°C, and dispersed using a homogenizer (ULTRA-TURRAX, manufactured by IKA Corporation) to obtain a crude liposome dispersion. The crude liposome dispersion was then dispersed using a high-pressure homogenizer (Nano-Mizer NM2, manufactured by Yoshida Kikai Kogyo Co., Ltd.) at 100 MPa to obtain liposomes with an average particle size (Z-average) of approximately 80 nm. Using a dialysis cassette (Slide-A-Lyzer G2 Dialysis Cassettes 20K MWCO, manufactured by Thermo Scientific), the outer aqueous phase of the liposomes was replaced with 10 mM L-histidine buffer / 10% sucrose solution (pH 6.5) to obtain a blank liposome solution. The mixture was filtered through a 0.22 μm membrane filter, and a 10 mM L-histidine buffer / 10% sucrose solution (pH 6.5) was added to adjust the total lipid concentration to 75 mM (75 μmol / mL) or 50 mM (50 μmol / mL). The amount of the mixture was adjusted as needed.

[0378] 2-3 mg of the test compound was weighed out, 1 mL of empty liposome solution with a total lipid concentration of 50 mM was added, and the pH was adjusted to 5.5-7 with 1 mol / L hydrochloric acid or 1 mol / L sodium hydroxide aqueous solution, and the mixture was heated in a water bath at 65°C for 10-30 minutes and then cooled on ice. Any insoluble matter was removed by centrifugation at 15,000 x g for 5 minutes.

[0379] The liposome encapsulation rate was calculated as follows. 100 μL of the liposome solution was placed in an ultrafiltration filter (Amicon Ultra, 100K, 0.5 mL, manufactured by Merck) and centrifuged at 4°C and 15,000 × g for 10 minutes. The compound concentration in the filtrate after ultrafiltration was measured by HPLC and used as the unencapsulated compound concentration. The liposome solution was diluted with a trifluoroacetic acid / water / methanol mixture (0.1 / 25 / 75) and allowed to stand at 5°C for 10 minutes or more. Insoluble matter was removed by centrifugation at 15,000 × g for 5 minutes, and the compound concentration in the supernatant was measured by HPLC and used as the compound concentration in the liposome solution. The encapsulation rate and encapsulation efficiency were calculated using the following formulas. Encapsulation rate (%) = (compound concentration in liposome solution - concentration of unencapsulated compound) x 100 / compound concentration in liposome solution Encapsulation efficiency (%) = (compound concentration in liposome solution - concentration of unencapsulated compound) x 100 / compound concentration at time of introduction

[0380] The HPLC measurement conditions are as follows: HPLC conditions Column: Acquity UPLC BEH C18, 1.7 μm, 50 × 2.1 mm Column temperature: 40°C Mobile phase: A: Water containing 0.1% trifluoroacetic acid B: Acetonitrile A / B (min): 95 / 5 (0) → 0 / 100 (3.5) → 0 / 100 (4) → 95 / 5 (4.01) → 95 / 5 (5) Flow rate: 0.8 mL / min Detection: UV-visible detector Measurement wavelength: 254 nm Injection volume: 3 μL or 5 μL

[0381]

[0382] It was confirmed that prexasertib and Examples 1, 3, 4, 5, and 6 achieved high encapsulation efficiencies of 80% or more. Furthermore, during the liposome encapsulation procedure in Example 4, a significant increase in liquid viscosity was observed when 1 mol / L hydrochloric acid was added to adjust the pH. This increase in viscosity may pose a problem when producing liposomes on a large scale. On the other hand, no increase in viscosity was observed during the liposome encapsulation procedure in Examples 1, 3, 5, and 6. Therefore, the compounds represented by formulas (3) and (4) demonstrated particularly favorable liposome encapsulation operability.

[0383] Test Example 6A. Liposome Encapsulation Test of Example 1 A liposome encapsulation test was carried out on the compound of Example 1.

[0384] 11.08 g of Presome ACD-1 (a pre-prepared mixture of hydrogenated soybean phosphatidylcholine, cholesterol, and distearoylphosphatidylethanolamine-methoxypolyethylene glycol 2000 in a mass ratio of 3:1:1, manufactured by Nippon Fine Chemical Co., Ltd.) was weighed out, 200 mL of 250 mM ammonium sulfate solution was added, and the mixture was heated to 65°C and dispersed using a homogenizer (ULTRA-TURRAX, manufactured by IKA Corporation) to obtain a crude liposome dispersion. The crude liposome dispersion was further dispersed using a high-pressure homogenizer (Nano-Mizer NM2, manufactured by Yoshida Kikai Kogyo Co., Ltd.) at a pressure of 100 MPa to obtain liposomes with an average particle size (Z-average) of 80 nm. Using a dialysis cassette (Slide-A-Lyzer G2 Dialysis Cassettes 20K MWCO, manufactured by Thermo Scientific), the outer aqueous phase of the liposomes was replaced with a 10 mM L-histidine buffer / 10% sucrose solution (pH 6.5), and the resulting solution was filtered through a 0.22 μm membrane filter to obtain a blank liposome solution with a total lipid concentration of 75 mM (75 μmol / mL).

[0385] The compound of Example 1 was weighed out, and about 0.4 mL of 10 mM L-histidine buffer / 10% sucrose solution (pH 6.5), about 0.1 mL of 0.1 mol / L hydrochloric acid, and 1 mL of the above empty liposome solution were added, and the pH was adjusted to 6 to 6.5 by adding 1 mol / L hydrochloric acid. The mixture was heated in a water bath at 65°C for 30 minutes and then cooled on ice. Thereafter, the mixture was filtered through a 0.22 μm membrane filter.

[0386] The liposome encapsulation rate was calculated as follows. Unencapsulated compound concentration: 100 μL of 4% phosphoric acid aqueous solution was added to 100 μL of liposome solution, and the compound concentration was measured by HPLC. The HPLC measurement conditions are as follows. HPLC conditions Column: MonoSelect nPEC (GL Sciences Inc.) Column temperature: 30°C Mobile phase: A: Water containing 0.1% trifluoroacetic acid B: Acetonitrile A / B (min): 95 / 5 (0) → 95 / 5 (1) → 70 / 30 (6) → 70 / 30 (7) → 95 / 5 (7.01) → 95 / 5 (10) Flow rate: 1 mL / min Detection: UV-visible detector Measurement wavelength: 254 nm Injection volume: 2 μL

[0387] Compound concentration in liposome solution: The liposome solution was diluted with a trifluoroacetic acid / water / methanol mixture (0.1 / 25 / 75) and allowed to stand at 5°C for 10 minutes or more. The mixture was centrifuged at 15,000 x g for 5 minutes to remove insoluble matter, and the compound concentration of the supernatant was measured by HPLC. The HPLC measurement conditions were as follows. HPLC conditions: Column: Acquity UPLC BEH C18, 1.7 μm, 50 x 2.1 mm; Column temperature: 40°C; Mobile phase: A: Water containing 0.1% trifluoroacetic acid; B: Acetonitrile; A / B (min): 95 / 5 (0) → 0 / 100 (3.5) → 0 / 100 (4) → 95 / 5 (4.01) → 95 / 5 (5); Flow rate: 0.8 mL / min; Detection: UV-visible detector; Measurement wavelength: 254 nm; Injection volume: 5 μL

[0388] The encapsulation rate and encapsulation efficiency were calculated using the following formulas: encapsulation rate (%) = (compound concentration in liposome solution - concentration of unencapsulated compound) x 100 / compound concentration in liposome solution encapsulation efficiency (%) = (compound concentration in liposome solution - concentration of unencapsulated compound) x 100 / compound concentration at introduction

[0389]

[0390] It was confirmed that Example 1 was able to achieve a high liposome encapsulation efficiency of 90% or more.

[0391] Test Example 6B. Liposome Encapsulation Test of Example 1 A liposome encapsulation test was carried out on the compound of Example 1.

[0392] 11.08 g of Presome ACD-1 (a pre-prepared mixture of hydrogenated soybean phosphatidylcholine, cholesterol, and distearoylphosphatidylethanolamine-methoxypolyethylene glycol 2000 in a mass ratio of 3:1:1, manufactured by Nippon Fine Chemical Co., Ltd.) was weighed out, 200 mL of 250 mM ammonium sulfate solution was added, and the mixture was heated to 65°C and dispersed using a homogenizer (ULTRA-TURRAX, manufactured by IKA Corporation) to obtain a crude liposome dispersion. The crude liposome dispersion was further dispersed using a high-pressure homogenizer (Nano-Mizer NM2, manufactured by Yoshida Kikai Kogyo Co., Ltd.) at a pressure of 100 MPa to obtain liposomes with an average particle size (Z-average) of 81 nm. Using a dialysis cassette (Slide-A-Lyzer G2 Dialysis Cassettes 20K MWCO, manufactured by Thermo Scientific), the outer aqueous phase of the liposomes was replaced with a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5), and the resulting solution was filtered through a 0.22 μm membrane filter to obtain a blank liposome solution with a total lipid concentration of 53.1 mg / mL.

[0393] 120 mg of the compound of Example 1 was weighed out, and 18.3 mL of 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5), 0.3 mL of 1 mol / L hydrochloric acid, and 41.7 mL of the above empty liposome solution were added, and the pH was adjusted to 6.5 with 1 mol / L hydrochloric acid or 1 mol / L sodium hydroxide. This solution was heated in a 50°C water bath for 30 minutes, then ice-cooled and filtered through a 0.22 μm membrane filter. The compound concentration in the liposome solution of Example 1 was 2.00 mg / mL, the encapsulation rate was 98.0%, and the average particle size (Z-average) was 84 nm.

[0394] The liposome encapsulation rate was calculated as follows. Unencapsulated compound concentration: 100 μL of liposome solution, 100 μL of 4% phosphoric acid aqueous solution, and 300 μL of physiological saline were mixed and centrifuged at 100,000 × g for 60 minutes to remove insoluble matter, and the compound concentration in the supernatant was measured by HPLC. Compound concentration in liposome solution: The liposome solution was diluted with a trifluoroacetic acid / water / methanol mixture (0.1 / 25 / 75) and allowed to stand at 5°C for 10 minutes or more. Insoluble matter was removed by centrifugation at 15,000 × g for 5 minutes, and the compound concentration in the supernatant was measured by HPLC. The HPLC measurement conditions are as follows. HPLC conditions Column: Acquity UPLC BEH C18, 1.7 μm, 50 × 2.1 mm Column temperature: 40 ° C. Mobile phase: A: Water containing 0.1% trifluoroacetic acid B: Acetonitrile A / B (min): 95 / 5 (0) → 0 / 100 (3.5) → 0 / 100 (4) → 95 / 5 (4.01) → 95 / 5 (5) Flow rate: 0.8 mL / min Detection: UV-visible detector Measurement wavelength: 254 nm Injection volume: 5 μL

[0395] The encapsulation rate was calculated using the following formula: encapsulation rate (%) = (compound concentration in liposome solution - unencapsulated compound concentration) x 100 / compound concentration in liposome solution

[0396] Liposomes encapsulating the compound of Example 1 were stored at 5° C., and changes in compound concentration, encapsulation rate, and average particle size were confirmed. As shown in Table 11, no significant changes were observed in the compound concentration, encapsulation rate, and average particle size of liposomes encapsulating Example 1, demonstrating excellent storage stability.

[0397]

[0398] Test Example 7 Pharmacokinetics Test The solution formulation of prexasertib, Examples 1, 3, and 6, and the liposome formulation of prexasertib were intravenously administered to mice, and the concentrations of the test substances in the blood were measured.

[0399] The solution preparations were prepared by dissolving the compound in a 10 mM glycine / 5% mannitol solution (pH 2) or a 10 mM glycine / 5% mannitol solution (pH 2) containing 20% ​​sulfobutylether-β-cyclodextrin, and then filtering the solution through a 0.22 μm membrane filter.

[0400] The liposome preparations used were prepared by preparing liposomes encapsulating a test compound in the same manner as in Test Example 6, replacing the outer aqueous phase of the liposomes with 10 mM L-histidine buffer / 10% sucrose solution (pH 6.5) using a gel filtration column PD-10 (GE Healthcare), filtering the liposomes with a 0.22 μm membrane filter, and adjusting the concentration by adding 10 mM L-histidine buffer / 10% sucrose solution (pH 6.5).

[0401] <Administration test> The solution formulation or liposome formulation was intravenously administered to 7-week-old female BALB / c mice, and blood was collected from the jugular vein without anesthesia over time up to 72 hours after administration. Immediately after collection, four volumes of methanol were added to the blood, which was then centrifuged, and the concentration of the test compound in the resulting supernatant was quantified by LC-MS / MS.

[0402] The LC-MS / MS measurement conditions were as follows: HPLC: Prominence system (Shimadzu Corporation) MS / MS: 4000 QTRAP (SCIEX) Column: Cadenza CD-C18, 3 μm, 50 × 2 mm (Intact Corporation) Column temperature: 40°C Mobile phase: A: water containing 0.1% formic acid B: acetonitrile containing 0.1% formic acid A / B (min): 90 / 10 (0) → 10 / 90 (2.5) → 10 / 90 (3.5) → 90 / 10 (3.6) → 90 / 10 (5.0) Flow rate: 0.4 mL / min Detection: ESI (positive mode) Injection volume: 0.1 to 5 μL

[0403] The test results are shown in the tables below. In the tables, "mean" means the average and "S.D." means the standard deviation. The plasma concentrations of the test compound from 0 hours after administration to 72 hours after administration are shown in Tables 12, 13, 14, 15, and 16.

[0404]

[0405]

[0406]

[0407]

[0408]

[0409] The AUC for Examples 1, 3, and 6 and the liposome formulation of prexasertib was calculated by the trapezoidal method from 0 hour after administration to the time point (t) at which the plasma test compound concentration could be quantified, and is shown in Table 17.

[0410] The above results confirmed that the liposome-encapsulated liposomes of Examples 1 and 3, when administered intravenously, exhibited high blood retention. These results demonstrated the achievement of a sustained exposure of 72 hours, which is important for maximizing the efficacy of prexasertib, as reported in non-clinical studies. Therefore, Examples 1 and 3 have excellent pharmacokinetic profiles and are highly useful as anticancer agents. It was demonstrated that the specific position of the nitrogen atom on the pyridine ring, a characteristic of the compound represented by formula (3), is important.

[0411] Test Example 8. Drug efficacy evaluation test using tumor-bearing mice transplanted with ES-2 cells. The antitumor effects of prexasertib, a liposome preparation encapsulating prexasertib, and a liposome preparation encapsulating Example 1 were evaluated.

[0412] The solution formulation of prexasertib was prepared by dissolving prexasertib in a 10 mM glycine / 5% mannitol solution (pH 2) containing 20% ​​sulfobutylether-β-cyclodextrin, and then filtering the solution through a 0.22 μm membrane filter.

[0413] Liposomal formulations were prepared in the same manner as in Test Example 6. A test compound was weighed out, and a blank liposome solution with a total lipid concentration of 50 mM was added to the mixture to give a compound concentration of 2 mg / mL. 1 mol / L hydrochloric acid was added to adjust the pH to 6-7. Alternatively, a 10 mM L-histidine buffer / 10% sucrose solution (pH 6.5) and hydrochloric acid were added to the test compound to dissolve or disperse the compound, and a blank liposome solution with a total lipid concentration of 75 mM was added to give a compound concentration of 2 mg / mL, adjusting the pH to 6-7. This solution was heated in a 65°C water bath for 10-30 minutes and then ice-cooled. The mixture was allowed to stand overnight or longer in a 5°C refrigerator and then filtered through a 0.22 μm membrane filter. The liposome encapsulation rate calculated by the method described in Test Example 6 was 99% or higher.

[0414] 4-7 week-old BALB / c-nu / nu mice (CANN.Cg-Foxn1nu / CrlCrlj, female, Charles River Japan) were inoculated with 1 × 10 ES-2 cells (ATCC). ６ The tumors were intradermally transplanted around the ventral region to achieve a ratio of 1000 cells / mouse. After confirming the engraftment of ES-2 cells 5 to 14 days after transplantation, the solution formulation was administered intravenously once a week at a dose of 30 mg / kg or the liposome formulation was administered subcutaneously twice a day for three consecutive days at a dose of 3 mg / kg, 7.5 mg / kg, or 20 mg / kg (a total of six administrations per week). The tumor volume was measured over time from the start of administration, and the effect of compound administration on tumor volume reduction was evaluated. The tumor volume was calculated using the following formula, using the minor and major diameters of the tumor measured with an electronic caliper (Mitutoyo).

[0415] Tumor volume [mm ３ ] = 0.5×(breadth diameter [mm]) ２ × major axis [mm]

[0416] A control group administered with only the solvent was compared with a group administered with a compound of the present disclosure, and the T / C was calculated using the following formula to evaluate the antitumor effect. The percentage of complete tumor regression (CR) individuals at the end of administration of the solution formulation or liposome formulation was also recorded. For the control group, an empty liposome solution prepared in the same manner as in Test Example 6 was used.

[0417] T / C (%) = (tumor volume at the end of administration in the group administered with the compound of the present disclosure - tumor volume at the start of administration in the group administered with the compound of the present disclosure) / (tumor volume at the end of administration in the control administration group - tumor volume at the start of administration in the control administration group) × 100

[0418] Table 18 shows the T / C (%) and CR rate for tumor-bearing mice transplanted with ES-2 cells at each dose and administration period of the test compound.

[0419] In efficacy evaluation tests using tumor-bearing mice, liposomally formulated Example 1 was found to exhibit excellent antitumor effects at all doses, similar to those of liposomally formulated plexasertib. In evaluation tests using a dose of 20 mg / kg, these liposomal formulations achieved tumor regression that was not achieved with solution-formulated plexasertib. These test results demonstrate that Example 1 is a promising compound that exhibits excellent antitumor effects and has exceptional efficacy.

[0420] Test Example 9. Measurement of neutrophil count using tumor-bearing mice transplanted with ES-2 cells 1×10 ES-2 cells (ATCC) were transplanted into 4- to 7-week-old BALB / c-nu / nu mice (CANN.Cg-Foxn1nu / CrlCrlj, female, Charles River Japan). ６ The cells were intradermally transplanted around the abdominal region to form a 100% ES-2 cell / mouse ratio. After confirming the engraftment of ES-2 cells 5 to 14 days after transplantation, a liposome formulation encapsulating prexasertib and a liposome formulation encapsulating Example 1 were administered intravenously in single doses. 72 hours after administration, blood was collected and the neutrophil count was measured. For comparison in this test, an empty liposome solution prepared in the same manner as in Test Example 6 was used.

[0421] A control group administered with blank liposome solution was compared with treatment groups administered with liposome-formulated Example 1 and liposome-formulated plexasertib, and the neutrophil survival rate was calculated using the following formula to evaluate the safety of each formulation.

[0422] Neutrophil survival rate (%) = (neutrophil count in the treatment group 72 hours after administration) / (neutrophil count in the control treatment group 72 hours after administration) × 100

[0423] Table 19 shows the survival rate of neutrophils in tumor-bearing mice transplanted with ES-2 cells.

[0424]

[0425] The liposomal formulation of Example 1 exhibits the same antitumor effect as liposomal formulation of plexasertib at the same dose. At the minimum effective dose of 3 mg / kg, liposomal formulation of plexasertib exhibited a neutrophil survival rate of 15%, demonstrating simultaneous observation of antitumor efficacy and hematotoxicity. On the other hand, at the minimum effective dose of 3 mg / kg, liposomal formulation of Example 1 exhibited a neutrophil survival rate of 61%, demonstrating reduced hematotoxicity side effects. Furthermore, as shown in Test Example 8, liposomal formulation of Example 1 exhibited a higher neutrophil survival rate at a dose of 20 mg / kg, at which tumor regression was observed, than liposomal formulation of plexasertib administered at 7.5 mg / kg. Thus, Example 1 is an excellent compound possessing both higher safety and efficacy than plexasertib, and exhibits exceptional effects. Similarly, the liposomal formulation of Example 1 is an excellent compound possessing both high safety and efficacy, and exhibits exceptional effects.

[0426] Test Example 10. Human bone marrow cell myeloid differentiation induction test (hematotoxicity evaluation test) Human bone marrow CD34-positive hematopoietic stem cells were obtained from Lonza Co., Ltd. The frozen cells were thawed in IMDM medium containing 1% fetal bovine serum, suspended in Complete hemaTox (registered trademark) Myeloid Medium, and seeded at 1,000 cells per well in a 96-well plate. The compound to be evaluated was added to a final DMSO concentration of 0.1% and 3 nM, and the cells were incubated at 37°C and 5% CO ２ After the completion of the culture, half of the cell suspension per well was collected, and the luminescence intensity was measured using CellTiter-Glo (registered trademark) 3D Reagent (Promega, G968B). The cell survival rate was calculated using the following formula.

[0427] Cell survival rate (%)=(luminescence measurement value of the compound to be evaluated after 7 days) / (luminescence measurement value of the control group after 7 days)×100

[0428] The representative compounds obtained in the Examples and Prexasertib were subjected to the test shown in Test Example 10. The results are shown in the table below.

[0429] As shown in the above table, it was revealed that although 3 nM of Prexasertib exhibits an inhibitory effect on myeloid cells differentiated from human bone marrow cells, 3 nM of Example 1 does not exhibit an inhibitory effect on myeloid cells differentiated from human bone marrow cells. From these results, compared to Prexasertib, Example 1 is a compound that is highly safe in terms of hematotoxicity and has exceptional and unique effects.

[0430] Test Example 11. Tumor PD test using tumor-bearing mice transplanted with ES-2 cells. The pharmacodynamic (PD) response in tumors was evaluated using prexasertib, a liposome preparation encapsulating prexasertib, and a liposome preparation encapsulating Example 1.

[0431] The solution formulation of prexasertib was prepared by dissolving prexasertib in a 10 mM glycine / 5% mannitol solution (pH 2) containing 20% ​​sulfobutylether-β-cyclodextrin, and then filtering the solution through a 0.22 μm membrane filter.

[0432] Liposomal formulations were prepared in the same manner as in Test Example 6. A test compound was weighed out, and a blank liposome solution with a total lipid concentration of 50 mM was added to the mixture to give a compound concentration of 2 mg / mL. 1 mol / L hydrochloric acid was added to adjust the pH to 6-7. Alternatively, a 10 mM L-histidine buffer / 10% sucrose solution (pH 6.5) and hydrochloric acid were added to the test compound to dissolve or disperse the compound, and a blank liposome solution with a total lipid concentration of 75 mM was added to give a compound concentration of 2 mg / mL, adjusting the pH to 6-7. This solution was heated in a 65°C water bath for 10-30 minutes and then ice-cooled. The mixture was allowed to stand overnight or longer in a 5°C refrigerator and then filtered through a 0.22 μm membrane filter. The liposome encapsulation rate calculated by the method described in Test Example 6 was 99% or higher.

[0433] 4-7 week old BALB / c-nu / nu mice (CANN.Cg-Foxn1nu / CrlCrlj, female, Charles River Japan) were inoculated with 1 × 10 ovarian cancer ES-2 cells (ATCC). ６ The tumors were intradermally transplanted around the ventral region to achieve a concentration of 1000 cells / mouse. Five to 14 days after transplantation, ES-2 cell engraftment was confirmed, and then the cells were administered under the following conditions for each test: Test A: A single intravenous administration of the solution formulation at 30 mg / kg or the liposome formulation at 7.5 mg / kg, 3 mg / kg, 1 mg / kg, or 0.3 mg / kg was performed. Test B: A single intravenous administration of the solution formulation at 30 mg / kg or the liposome formulation at 7.5 mg / kg, 3 mg / kg, 1 mg / kg, or 0.3 mg / kg was performed. Test C: A single intravenous administration of the liposome formulation at 20 mg / kg, 7.5 mg / kg, 3 mg / kg, 1 mg / kg, or 0.3 mg / kg was performed. Tumors were harvested 1, 3, or 6 days after administration, and the PD response of the tumors to the compound administration was evaluated.

[0434] The PD response in tumors was evaluated by Western blotting. Band intensities detected with anti-γH2AX antibody (05-636, Merck) and anti-tubulin antibody (3873, Cell Signaling Technology) were calculated using ImageJ software.

[0435] A control group administered with only the solvent was compared with a group administered with the compound of the present disclosure, and the relative value of each band intensity was calculated using the following formula to evaluate the PD response effect in the tumor. For the control group, an empty liposome solution prepared in the same manner as in Test Example 6 was used.

[0436] γH2AX intensity={(γH2AX band intensity in tumors of the group administered with the compound of the present disclosure) / (Tubulin band intensity in tumors of the group administered with the compound of the present disclosure)} / {(γH2AX band intensity in tumors of the control group / Tubulin band intensity in tumors of the control group)}×100

[0437] 7 to 9 show the intensity of γH2AX in tumor-bearing mice transplanted with ES-2 cells at various doses and administration periods of the test compound.

[0438]

[0073] From the above results, as shown in Test Example 11, Test A, when liposomal formulation of plexasertib was administered, a strong PD response was observed at a dose of 7.5 mg / kg three days after administration, achieving a PD response that could not be achieved with solution-formulated plexasertib. As shown in Test Example 11, Test B, when liposomal formulation of Example 1 was administered, a strong PD response was observed at a dose of 7.5 mg / kg one day after administration, achieving a PD response that could not be achieved with solution-formulated plexasertib. As shown in Test Example 11, Test C, when liposomal formulation of Example 1 was administered, a strong PD response was observed at doses of 7.5 mg / kg and 20 mg / kg three and six days after administration. As shown in Test Example 7, these liposomal formulations achieved the 72-hour sustained exposure that is important for maximizing the efficacy of plexasertib, as reported in non-clinical studies. Therefore, Example 1 has excellent pharmacokinetic and pharmacodynamic profiles and is extremely useful as an anticancer agent.

[0439] Test Example 12. Drug efficacy evaluation test using peritoneally disseminated cancer-bearing mice transplanted with ES-2 cells. The antitumor effects were evaluated using prexasertib, a liposome preparation encapsulating prexasertib, and a liposome preparation encapsulating Example 1.

[0440] The solution formulation of prexasertib was prepared by dissolving prexasertib in a 10 mM glycine / 5% mannitol solution (pH 2) containing 20% ​​sulfobutylether-β-cyclodextrin, and then filtering the solution through a 0.22 μm membrane filter.

[0441] A liposome formulation encapsulating prexasertib was prepared as follows: 11.08 g of Presome ACD-1 (Nippon Fine Chemicals Co., Ltd.) and 200 mL of 250 mM ammonium sulfate solution were mixed and dispersed using a homogenizer (ULTRA-TURRAX, IKA Corporation) to obtain a crude liposome dispersion. The crude liposome dispersion was further dispersed using a high-pressure homogenizer (Nano-Mizer NM2, Yoshida Kikai Kogyo Co., Ltd.) at a pressure of 100 MPa to obtain liposomes with an average particle size (Z-average) of 80 nm. Using a dialysis cassette (Slide-A-Lyzer G2 Dialysis Cassettes 20K MWCO, Thermo Scientific), the outer aqueous phase of the liposomes was replaced with a 10 mM L-histidine buffer / 10% sucrose solution (pH 6.5) and filtered through a 0.22 μm membrane filter to obtain a blank liposome solution with a total lipid concentration of 75 mM. 60 mg of plexasertib, 10 mL of a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5), and 20 mL of the blank liposome solution were added and adjusted to pH 6.5. This solution was heated in a 50°C water bath for 30 minutes, then cooled on ice and filtered through a 0.22 μm membrane filter. The concentration of plexasertib in the liposome solution was 1.97 mg / mL, and the encapsulation rate was 98%. The compound concentration and encapsulation rate in the liposome solution were calculated as follows. Unencapsulated compound concentration: 100 μL of liposome solution, 100 μL of 4% phosphoric acid aqueous solution, and 300 μL of physiological saline were mixed and centrifuged at 100,000 × g for 60 minutes to remove insoluble matter, and the compound concentration in the supernatant was measured by HPLC. Compound concentration in liposome solution: The liposome solution was diluted with a trifluoroacetic acid / water / methanol mixture (0.1 / 25 / 75), centrifuged at 4°C and 15,000 × g for 10 minutes to remove insoluble matter, and the compound concentration in the supernatant was measured by HPLC. The HPLC measurement conditions were as follows.HPLC conditions Column: Acquity UPLC BEH C18, 1.7 μm, 50 × 2.1 mm Column temperature: 40°C Mobile phase: A: Water containing 0.1% trifluoroacetic acid B: Acetonitrile A / B (min): 90 / 10 (0) → 70 / 30 (3) → 0 / 100 (3.5) → 0 / 100 (4) → 90 / 10 (4.01) → 90 / 10 (5) Flow rate: 0.8 mL / min Detection: UV-visible detector Measurement wavelength: 254 nm Injection volume: 5 μL The encapsulation rate was calculated using the following formula: Encapsulation rate (%) = (concentration of compound in liposome solution - concentration of unencapsulated compound) × 100 / concentration of compound in liposome solution.

[0442] A liposome formulation encapsulating Example 1 was prepared as follows. 11.08 g of Presome ACD-1 (manufactured by Nippon Fine Chemical Co., Ltd.) and 200 mL of 250 mM ammonium sulfate solution were mixed and dispersed using a homogenizer (ULTRA-TURRAX, manufactured by IKA Corporation) to obtain a crude liposome dispersion. The crude liposome dispersion was further dispersed using a high-pressure homogenizer (Nano-Mizer NM2, manufactured by Yoshida Kikai Kogyo Co., Ltd.) at a pressure of 100 MPa to obtain liposomes with an average particle size (Z-average) of 81 nm. Using a dialysis cassette (Slide-A-Lyzer G2 Dialysis Cassettes 20K MWCO, Thermo Scientific), the outer aqueous phase of the liposomes was replaced with a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5), and filtered through a 0.22 μm membrane filter to obtain a blank liposome solution with a total lipid concentration of 53.1 mg / mL. 80 mg of the compound of Example 1, 12.2 mL of a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5), and 27.8 mL of the blank liposome solution were added, and the pH was adjusted to 6.5. This solution was heated in a 50°C water bath for 30 minutes, then cooled on ice, and filtered through a 0.22 μm membrane filter. The compound concentration in the liposome solution in Example 1 was 1.99 mg / mL, and the encapsulation rate was 97%. The liposome encapsulation rate was calculated as follows. Unencapsulated compound concentration: 100 μL of 4% phosphoric acid aqueous solution was added to 100 μL of liposome solution, and the compound concentration was measured by HPLC. The HPLC measurement conditions were as follows. HPLC Conditions Column: MonoSelect nPEC (GL Sciences Inc.) Column Temperature: 30°C Mobile Phase: A: 0.1% trifluoroacetic acid-containing water B: acetonitrile A / B (min): 95 / 5 (0) → 95 / 5 (1) → 70 / 30 (6) → 70 / 30 (7) → 95 / 5 (7.01) → 95 / 5 (10) Flow Rate: 1 mL / min Detection: UV-Visible Detector Measurement Wavelength: 254 nm Injection Volume: 2 μL Compound Concentration in Liposome Solution: The liposome solution was diluted with a trifluoroacetic acid / water / methanol mixture (0.1 / 25 / 75), centrifuged at 4°C and 15,000 × g for 10 minutes to remove insoluble matter, and the compound concentration of the supernatant was measured by HPLC. The HPLC measurement conditions are as follows.HPLC conditions Column: Acquity UPLC BEH C18, 1.7 μm, 50 × 2.1 mm Column temperature: 40°C Mobile phase: A: Water containing 0.1% trifluoroacetic acid B: Acetonitrile A / B (min): 95 / 5 (0) → 0 / 100 (3.5) → 0 / 100 (4) → 95 / 5 (4.01) → 95 / 5 (5) Flow rate: 0.8 mL / min Detection: UV-visible detector Measurement wavelength: 254 nm Injection volume: 5 μL The encapsulation rate was calculated using the following formula: Encapsulation rate (%) = (compound concentration in liposome solution - concentration of unencapsulated compound) × 100 / compound concentration in liposome solution.

[0443] Five-week-old BALB / c-nu / nu mice (CAnN.Cg-Foxn1nu / CrlCrlj, female, Charles River Japan) were inoculated with 1 × 10 ovarian cancer ES-2 cells (ATCC) expressing luciferase. ６ The cells were transplanted intraperitoneally to form a 100% ES-2 cell / mouse ratio. Five to 14 days after transplantation, the engraftment of ES-2 cells was confirmed using an IVIS Imaging System (PerkinElmer), and then the solution formulation was administered intravenously once a week at a dose of 30 mg / kg or the liposome formulation at 4.5 mg / kg or 20 mg / kg. Luciferase-induced luminescence was measured over time from the start of administration to evaluate the tumor shrinkage effect of compound administration. The mice were observed based on the tumor size, systemic symptoms, and weight transition estimated from luciferase-induced luminescence, and the survival time of mice that showed no serious findings and were in good health was measured.

[0444] The median survival time was calculated for the control group administered with only the solvent and the group administered with the compound of the present disclosure, and the antitumor effect was evaluated. An empty liposome solution was used for the control group.

[0445] Table 21 shows the median survival time for mice bearing peritoneal disseminated ES-2 cell transplanted cancer at each dose and administration period of the test compound.

[0446] In a pharmacological efficacy evaluation test using mice bearing peritoneal disseminated cancer, it was revealed that liposomally formulated Example 1 exhibited excellent antitumor effects at a dose of 20 mg / kg, similar to liposomally formulated prexasertib. In an evaluation test using a dose of 20 mg / kg, these liposomal formulations achieved an extension of survival time that could not be achieved with solution-formulated prexasertib. The results of this test demonstrate that Example 1 is a promising compound that exhibits excellent antitumor effects and has exceptional efficacy.

[0447] Test Example 13. Drug efficacy evaluation test using ovarian orthotopic cancer-bearing mice transplanted with ES-2 cells. The antitumor effects of prexasertib, a liposome preparation encapsulating prexasertib, and a liposome preparation encapsulating Example 1 were evaluated.

[0448] The solution formulation of prexasertib was prepared by dissolving prexasertib in a 10 mM glycine / 5% mannitol solution (pH 2) containing 20% ​​sulfobutylether-β-cyclodextrin, and then filtering the solution through a 0.22 μm membrane filter.

[0449] A liposome formulation encapsulating prexasertib was prepared as follows: 11.08 g of Presome ACD-1 (Nippon Fine Chemicals Co., Ltd.) and 200 mL of 250 mM ammonium sulfate solution were mixed and dispersed using a homogenizer (ULTRA-TURRAX, IKA Corporation) to obtain a crude liposome dispersion. The crude liposome dispersion was further dispersed using a high-pressure homogenizer (Nano-Mizer NM2, Yoshida Kikai Kogyo Co., Ltd.) at a pressure of 100 MPa to obtain liposomes with an average particle size (Z-average) of 80 nm. Using a dialysis cassette (Slide-A-Lyzer G2 Dialysis Cassettes 20K MWCO, Thermo Scientific), the outer aqueous phase of the liposomes was replaced with a 10 mM L-histidine buffer / 10% sucrose solution (pH 6.5) and filtered through a 0.22 μm membrane filter to obtain a blank liposome solution with a total lipid concentration of 75 mM. 60 mg of plexasertib, 10 mL of a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5), and 20 mL of the blank liposome solution were added and adjusted to pH 6.5. This solution was heated in a 50°C water bath for 30 minutes, then cooled on ice and filtered through a 0.22 μm membrane filter. The concentration of plexasertib in the liposome solution was 1.97 mg / mL, and the encapsulation rate was 98%. The compound concentration and encapsulation rate in the liposome solution were calculated as follows. Unencapsulated compound concentration: 100 μL of liposome solution, 100 μL of 4% phosphoric acid aqueous solution, and 300 μL of physiological saline were mixed and centrifuged at 100,000 × g for 60 minutes to remove insoluble matter, and the compound concentration in the supernatant was measured by HPLC. Compound concentration in liposome solution: The liposome solution was diluted with a trifluoroacetic acid / water / methanol mixture (0.1 / 25 / 75), centrifuged at 4°C and 15,000 × g for 10 minutes to remove insoluble matter, and the compound concentration in the supernatant was measured by HPLC. The HPLC measurement conditions were as follows.HPLC conditions Column: Acquity UPLC BEH C18, 1.7 μm, 50 × 2.1 mm Column temperature: 40°C Mobile phase: A: Water containing 0.1% trifluoroacetic acid B: Acetonitrile A / B (min): 90 / 10 (0) → 70 / 30 (3) → 0 / 100 (3.5) → 0 / 100 (4) → 90 / 10 (4.01) → 90 / 10 (5) Flow rate: 0.8 mL / min Detection: UV-visible detector Measurement wavelength: 254 nm Injection volume: 5 μL The encapsulation rate was calculated using the following formula: Encapsulation rate (%) = (concentration of compound in liposome solution - concentration of unencapsulated compound) × 100 / concentration of compound in liposome solution.

[0450] A liposome formulation encapsulating Example 1 was prepared as follows. 11.08 g of Presome ACD-1 (manufactured by Nippon Fine Chemical Co., Ltd.) and 200 mL of 250 mM ammonium sulfate solution were mixed and dispersed using a homogenizer (ULTRA-TURRAX, manufactured by IKA Corporation) to obtain a crude liposome dispersion. The crude liposome dispersion was further dispersed using a high-pressure homogenizer (Nano-Mizer NM2, manufactured by Yoshida Kikai Kogyo Co., Ltd.) at a pressure of 100 MPa to obtain liposomes with an average particle size (Z-average) of 81 nm. Using a dialysis cassette (Slide-A-Lyzer G2 Dialysis Cassettes 20K MWCO, Thermo Scientific), the outer aqueous phase of the liposomes was replaced with a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5), and filtered through a 0.22 μm membrane filter to obtain a blank liposome solution with a total lipid concentration of 53.1 mg / mL. 80 mg of the compound of Example 1, 12.2 mL of a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5), and 27.8 mL of the blank liposome solution were added, and the pH was adjusted to 6.5. This solution was heated in a 50°C water bath for 30 minutes, then cooled on ice, and filtered through a 0.22 μm membrane filter. The compound concentration in the liposome solution in Example 1 was 1.99 mg / mL, and the encapsulation rate was 97%. The liposome encapsulation rate was calculated as follows. Unencapsulated compound concentration: 100 μL of 4% phosphoric acid aqueous solution was added to 100 μL of liposome solution, and the compound concentration was measured by HPLC. The HPLC measurement conditions were as follows. HPLC Conditions Column: MonoSelect nPEC (GL Sciences Inc.) Column Temperature: 30°C Mobile Phase: A: 0.1% trifluoroacetic acid-containing water B: acetonitrile A / B (min): 95 / 5 (0) → 95 / 5 (1) → 70 / 30 (6) → 70 / 30 (7) → 95 / 5 (7.01) → 95 / 5 (10) Flow Rate: 1 mL / min Detection: UV-Visible Detector Measurement Wavelength: 254 nm Injection Volume: 2 μL Compound Concentration in Liposome Solution: The liposome solution was diluted with a trifluoroacetic acid / water / methanol mixture (0.1 / 25 / 75), centrifuged at 4°C and 15,000 × g for 10 minutes to remove insoluble matter, and the compound concentration of the supernatant was measured by HPLC. The HPLC measurement conditions are as follows.HPLC conditions Column: Acquity UPLC BEH C18, 1.7 μm, 50 × 2.1 mm Column temperature: 40°C Mobile phase: A: Water containing 0.1% trifluoroacetic acid B: Acetonitrile A / B (min): 95 / 5 (0) → 0 / 100 (3.5) → 0 / 100 (4) → 95 / 5 (4.01) → 95 / 5 (5) Flow rate: 0.8 mL / min Detection: UV-visible detector Measurement wavelength: 254 nm Injection volume: 5 μL The encapsulation rate was calculated using the following formula: Encapsulation rate (%) = (compound concentration in liposome solution - concentration of unencapsulated compound) × 100 / compound concentration in liposome solution.

[0451] Five-week-old BALB / c-nu / nu mice (CAnN.Cg-Foxn1nu / CrlCrlj, female, Charles River Japan) were inoculated with 1 × 10 ovarian cancer ES-2 cells (ATCC) expressing luciferase. ６ The cells were transplanted into the right ovary so as to form 1000 cells / mouse. Five to 14 days after transplantation, the engraftment of ES-2 cells was confirmed using an IVIS Imaging System (PerkinElmer), and then the solution formulation was administered intravenously once a week at a dose of 30 mg / kg or the liposome formulation at 4.5 mg / kg or 20 mg / kg. Luciferase-induced luminescence was measured over time from the start of administration to evaluate the tumor shrinkage effect of compound administration. The mice were observed based on the tumor size, systemic symptoms, and weight transition estimated from luciferase-induced luminescence, and the survival time of mice that showed no serious findings and were in good health was measured.

[0452] The median survival time was calculated for the control group administered with only the solvent and the group administered with the compound of the present disclosure, and the antitumor effect was evaluated. An empty liposome solution was used for the control group.

[0453] Table 22 shows the median survival time for ovarian orthotopic cancer-bearing mice transplanted with ES-2 cells at each dose and administration period of the test compound.

[0454] In a pharmacological efficacy evaluation test using mice bearing orthotopic ovarian cancer, it was revealed that liposomally formulated Example 1 exhibited excellent antitumor effects at a dose of 20 mg / kg, similar to liposomally formulated prexasertib. In evaluation tests using a dose of 20 mg / kg, these liposomal formulations achieved an extension of survival time that was not achieved with solution-formulated prexasertib. The results of this test demonstrate that Example 1 is a promising compound that exhibits excellent antitumor effects and has exceptional efficacy.

[0455] Test Example 14. Drug efficacy evaluation test using cancer-bearing mice transplanted with various cancer cells. The antitumor effect was evaluated using a liposome preparation encapsulating Example 1.

[0456] The liposome preparation encapsulating Example 1 was the same as that used in Test Example 12 or one prepared by the following method. 138.4 g of Presome ACD-1 (manufactured by Nippon Fine Chemical Co., Ltd.) was weighed out, and 2437 g of 250 mM ammonium sulfate solution was added. The mixture was dispersed using a homogenizer (CLEARMIX, manufactured by M TECHNIQUE) and degassed by vacuum degassing to obtain a crude liposome dispersion. The crude liposome dispersion was further dispersed using a high-pressure homogenizer (Microfluidizer M-110 EH-30, manufactured by Microfluidics) at a pressure of 14,500 psi to obtain liposomes with an average particle size (Z-average) of 73 nm. Using a tangential flow filtration system equipped with five Pellicon cassettes (Pellicon 2 Mini 300 kD 0.1 m², Merck), the liposome outer aqueous phase was replaced with a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5) and filtered through a 0.2 μm membrane filter to obtain a blank liposome solution with a total lipid concentration of 58.2 mg / mL. 3.0 g of the compound of Example 1 was weighed out, and 562 g of a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5) and 980 g of the blank liposome solution were added to adjust the pH to 6.5. The solution was heated at 45°C for 60 minutes and then cooled. Using a Tangential Flow Filtration system equipped with five Pellicon cassettes (Pellicon 2 Mini 300 kD 0.1 m², manufactured by Merck), the outer aqueous phase of the liposomes was replaced with a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5) and filtered through a 0.2 μm membrane filter. 50 mL of this solution was dispensed into glass vials, which were stoppered and seamed. The compound concentration in the liposome solution of Example 1 was 1.98 mg / mL, and the encapsulation rate was 97%. The compound concentration and encapsulation rate in the liposome solution were calculated as follows. Concentration of unencapsulated compound: 100 μL of liposome solution, 100 μL of 4% aqueous phosphoric acid solution, and 300 μL of physiological saline were mixed and centrifuged at 100,000×g for 60 minutes to remove insoluble matter, and the compound concentration in the supernatant was measured by HPLC.Compound concentration in liposome solution: The liposome solution was diluted with a trifluoroacetic acid / water / methanol mixture (0.1 / 25 / 75), centrifuged at 15,000×g at 4° C. for 10 minutes to remove insoluble matter, and the compound concentration in the supernatant was measured by HPLC under the following HPLC measurement conditions. HPLC conditions Column: Acquity UPLC BEH C18, 1.7 μm, 50 × 2.1 mm Column temperature: 40°C Mobile phase: A: Water containing 0.1% trifluoroacetic acid B: Acetonitrile A / B (min): 95 / 5 (0) → 60 / 40 (3) → 0 / 100 (3.5) → 0 / 100 (4) → 95 / 5 (4.01) → 95 / 5 (5) Flow rate: 0.8 mL / min Detection: UV-visible detector Measurement wavelength: 254 nm Injection volume: 5 μL The encapsulation rate was calculated using the following formula. Encapsulation rate (%) = (concentration of compound in liposome solution - concentration of unencapsulated compound) x 100 / concentration of compound in liposome solution Or, 11.08 g of Presome ACD-1 (manufactured by Nippon Fine Chemical Co., Ltd.) and 200 mL of 250 mM ammonium sulfate solution were mixed and dispersed using a homogenizer (ULTRA-TURRAX, manufactured by IKA Corporation) to obtain a crude liposome dispersion. The crude liposome dispersion was further dispersed using a high-pressure homogenizer (Nano-Mizer NM2, manufactured by Yoshida Kikai Kogyo Co., Ltd.) at a pressure of 100 MPa to obtain liposomes with an average particle size (Z-average) of 81 nm. Using a dialysis cassette (Slide-A-Lyzer G2 Dialysis Cassettes 20K MWCO, Thermo Scientific), the outer aqueous phase of the liposomes was replaced with a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5), and filtered through a 0.22 μm membrane filter to obtain a blank liposome solution with a total lipid concentration of 53.1 mg / mL. 60 mg of the compound of Example 1, 9.16 mL of a 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5), and 20.8 mL of the blank liposome solution were added, and the pH was adjusted to 6.5. This solution was heated in a 50°C water bath for 30 minutes, then cooled on ice, and filtered through a 0.22 μm membrane filter. The compound concentration in the liposome solution was 2.08 mg / mL and the encapsulation rate was 98% in Example 1. The compound concentration and encapsulation rate in the liposome solution were calculated as follows.Unencapsulated compound concentration: 100 μL of liposome solution, 100 μL of 4% phosphoric acid aqueous solution, and 300 μL of physiological saline were mixed and centrifuged at 100,000 × g for 60 minutes to remove insoluble matter, and the compound concentration in the supernatant was measured by HPLC. Compound concentration in liposome solution: The liposome solution was diluted with a trifluoroacetic acid / water / methanol mixture (0.1 / 25 / 75), centrifuged at 4°C and 15,000 × g for 10 minutes to remove insoluble matter, and the compound concentration in the supernatant was measured by HPLC. The HPLC measurement conditions were as follows. HPLC conditions Column: Acquity UPLC BEH C18, 1.7 μm, 50 × 2.1 mm Column temperature: 40°C Mobile phase: A: Water containing 0.1% trifluoroacetic acid B: Acetonitrile A / B (min): 95 / 5 (0) → 0 / 100 (3.5) → 0 / 100 (4) → 95 / 5 (4.01) → 95 / 5 (5) Flow rate: 0.8 mL / min Detection: UV-visible detector Measurement wavelength: 254 nm Injection volume: 5 μL The encapsulation rate was calculated using the following formula: Encapsulation rate (%) = (compound concentration in liposome solution - concentration of unencapsulated compound) × 100 / compound concentration in liposome solution.

[0457] BALB / c-nu / nu mice (CANN.Cg-Foxn1nu / CrlCrlj, female, Charles River, Japan) or NODSCID mice (NOD.CB17-Prkdc ｓｃｉｄ ovarian cancer SKOV-3 cells (ATCC), sarcoma SJCRH30 cells (ATCC), sarcoma HT-1080 cells (ATCC), pancreatic cancer AsPC-1 cells (ATCC), pancreatic cancer BxPC-3 cells (ATCC), lung cancer Calu-6 cells (ATCC), or ovarian cancer OV5304 cells (Crown Bioscience) were inoculated into a 5x10 ５ cells / mouse or 1 x 10 ６ cells / mouse or 3 x 10 ６ cells / mouse or 5 x 10 ６ cells / mouse or 8mm ３ ~27mm ３The tumors were intradermally transplanted around the ventral region to form a tumor mass per mouse. After confirming the engraftment of various cancer cells 5 to 27 days after transplantation, the liposome formulation was administered intravenously once a week at a dose of 4.5 mg / kg, 10 mg / kg, or 20 mg / kg. The tumor volume was measured over time from the start of administration, and the effect of compound administration on tumor volume reduction was evaluated. The tumor volume was calculated using the short and long diameters of the tumor measured with an electronic caliper (Mitutoyo) using the following formula. It is known that BxPC-3 cells are resistant to gemcitabine, and OV5304 is resistant to PARP inhibitors.

[0458] Tumor volume [mm ３ ] = 0.5×(breadth diameter [mm]) ２ × major axis [mm]

[0459] A control group administered with only the solvent was compared with a group administered with the compound of the present disclosure, and the T / C was calculated using the following formula to evaluate the antitumor effect. An empty liposome solution was used for the control group.

[0460] T / C (%) = (tumor volume at the end of administration in the group administered with the compound of the present disclosure - tumor volume at the start of administration in the group administered with the compound of the present disclosure) / (tumor volume at the end of administration in the control administration group - tumor volume at the start of administration in the control administration group) × 100

[0461] Table 23 shows the T / C (%) of the test compound at each dose and administration period in cancer-bearing mice transplanted with various cancer cells.

[0462] In a combined efficacy evaluation test using cancer-bearing mice, it was revealed that Example 1 formulated in liposomes exhibited excellent antitumor effects against various cancer cell lines. The test results demonstrate that Example 1 is a promising compound that exhibits excellent antitumor effects and has exceptional effects. Test Example 14A. Pharmacological efficacy evaluation test using cancer-bearing mice transplanted with various cancer cells The antitumor effect was evaluated using a liposome formulation encapsulating Example 1.

[0463] A liposome formulation encapsulating Example 1 was prepared as follows. 498.2 g of Presome ACD-1 (manufactured by Nippon Fine Chemical Co., Ltd.) was weighed out, and 8772 g of 250 mM ammonium sulfate solution was added. The mixture was dispersed using a homogenizer (CLEARMIX, manufactured by M TECHNIQUE) and degassed by vacuum degassing to obtain a crude liposome dispersion. The crude liposome dispersion was further dispersed using a high-pressure homogenizer (Microfluidizer M-110 EH-30, manufactured by Microfluidics) at a pressure of 14,500 psi to obtain liposomes with an average particle size (Z-average) of 73 nm. Using a Tangential Flow Filtration system equipped with five Pellicon cassettes (Pellicon 2 Mini 300 kD 0.1 m, manufactured by Merck), the outer aqueous phase of the liposomes was replaced with water for injection, and then L-histidine and sucrose were added to make a 10 mM L-histidine / 9.4% sucrose solution. The pH was adjusted to 6.5 with 1 mol / L hydrochloric acid, and the solution was filtered through a 0.2 μm membrane filter to obtain a blank liposome solution with a total lipid concentration of 50.2 mg / mL.

[0464] 10.0 g of the compound of Example 1 was weighed out, and 1354 g of 10 mM L-histidine buffer / 9.4% sucrose solution (pH 6.5), 27 g of 1 mol / L hydrochloric acid, and 3786 g of the empty liposome solution were added, and the pH was adjusted to 6.5 with 1 mol / L hydrochloric acid. After heating at 45°C for 60 minutes, the mixture was cooled, adjusted to pH 6.5, and filtered through a 0.2 μm membrane filter. This solution was dispensed in 50 mL aliquots into glass vials, stoppered, and seamed. The concentration of the compound of Example 1 in the liposome solution was 2.01 mg / mL, the encapsulation rate was 97.5%, and the average particle size (Z-average) was 74 nm.

[0465] The compound concentration and encapsulation rate in the liposome solution were calculated as follows: Unencapsulated compound concentration: 100 μL of liposome solution, 100 μL of 4% phosphoric acid aqueous solution, and 300 μL of physiological saline were mixed and centrifuged at 100,000 × g for 60 minutes to remove insoluble matter, and the compound concentration in the supernatant was measured by HPLC.

[0466] Compound concentration in liposome solution: The liposome solution was diluted with a trifluoroacetic acid / water / methanol mixture (0.1 / 25 / 75), centrifuged at 15,000×g at 4° C. for 10 minutes to remove insoluble matter, and the compound concentration in the supernatant was measured by HPLC under the following HPLC measurement conditions. HPLC conditions Column: Acquity UPLC BEH C18, 1.7 μm, 50 × 2.1 mm Column temperature: 40°C Mobile phase: A: Water containing 0.1% trifluoroacetic acid B: Acetonitrile A / B (min): 95 / 5 (0) → 60 / 40 (3) → 0 / 100 (3.5) → 0 / 100 (4) → 95 / 5 (4.01) → 95 / 5 (5) Flow rate: 0.8 mL / min Detection: UV-visible detector Measurement wavelength: 254 nm Injection volume: 5 μL The encapsulation rate was calculated using the following formula: Encapsulation rate (%) = (concentration of compound in liposome solution - concentration of unencapsulated compound) × 100 / concentration of compound in liposome solution

[0467] Four to seven-week-old BALB / c-nu / nu mice (CAnN.Cg-Foxn1nu / CrlCrlj, female, Charles River Japan) were inoculated with 1x10 A2780 cells (ECACC), HCC1806 cells (ATCC), An3CA cells (ATCC), Detroit 562 cells (ATCC), FaDu cells (ATCC), or NCI-H460 cells (ATCC). ６ cells / mouse or 2x10 ６ cells / mouse or 1.2x10 ６ cells / mouse or 1x10 ６ cells / mouse or 3x10 ６ cells / mouse or 3x10 ６ The tumors were intradermally transplanted around the ventral region to form 100 cells / mouse. Af...

Claims

1. A therapeutic and / or prophylactic agent for cancer patients who are resistant to immune checkpoint inhibitors, comprising a CHK1 inhibitor or liposomes encapsulating a CHK1 inhibitor as an active ingredient.

2. The therapeutic and / or prophylactic agent according to claim 1, characterized by using the CHK1 inhibitor in combination with an immune checkpoint inhibitor.

3. The therapeutic and / or prophylactic agent according to claim 2, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously.

4. The therapeutic and / or prophylactic agent according to claim 2, characterized in that the immune checkpoint inhibitor is administered after the administration of the CHK1 inhibitor.

5. The therapeutic and / or prophylactic agent according to claim 2, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously, followed by the administration of the anti-PD-1 antibody.

6. The therapeutic and / or prophylactic agent according to claim 2, characterized in that the CHK1 inhibitor and the immune checkpoint inhibitor are administered simultaneously, and then the anti-PD-1 antibody is administered three days or more later.

7. The therapeutic and / or prophylactic agent according to claim 2, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody or an anti-PD-L1 antibody.

8. The therapeutic and / or prophylactic agent according to claim 2, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody.

9. The therapeutic and / or prophylactic agent according to claim 2, characterized in that the anti-PD-1 antibody is pembrolizumab, nivolumab, spartalizumab, or semiprimab.

10. The therapeutic and / or prophylactic agent according to claim 1, characterized in that the number of CD3-positive cells in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

11. The therapeutic and / or prophylactic agent according to claim 1, characterized in that the number of CD8-positive cells in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

12. The therapeutic and / or prophylactic agent according to claim 1, characterized in that the number of CD8-positive cells in the tumor tissue of the cancer patient is 0.135 or less.

13. The therapeutic and / or prophylactic agent according to claim 1, characterized in that the M1-M2 ratio of macrophages in the tumor tissue of the cancer patient is 21 or less.

14. The therapeutic and / or prophylactic agent according to claim 1, characterized in that the number of CD206-positive cells in the tumor tissue of the cancer patient is increased compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

15. The therapeutic and / or prophylactic agent according to claim 1, characterized in that the number of IAd (MHC class II) positive cells in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

16. The therapeutic and / or prophylactic agent according to claim 1, characterized in that the number of CD62L-negative CD44-positive cells in the lymph nodes of the cancer patient is reduced compared to the lymph nodes of a patient responsive to anti-PD-1 antibodies.

17. The therapeutic and / or prophylactic agent according to claim 1, characterized in that PD-L1 expression in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of a patient responsive to an anti-PD-1 antibody.

18. The therapeutic and / or prophylactic agent according to claim 1, characterized in that VEGFa expression in the tumor tissue of the cancer patient is increased compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

19. The therapeutic and / or prophylactic agent according to claim 1, characterized in that Granzyme B expression in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

20. The therapeutic and / or prophylactic agent according to claim 1, characterized in that Interferon γ expression in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

21. The therapeutic and / or prophylactic agent according to claim 1, characterized in that the expression of H2ab1 (MHC class II) in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

22. The therapeutic and / or prophylactic agent according to claim 1, characterized in that CD80 expression in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

23. The therapeutic and / or prophylactic agent according to claim 1, characterized in that Mrc1 expression in the tumor tissue of the cancer patient is increased compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

24. The therapeutic and / or prophylactic agent according to claim 1, characterized in that Arginase1 expression in the tumor tissue of the cancer patient is increased compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

25. The therapeutic and / or prophylactic agent according to claim 1, characterized in that the DNA replication stress in the tumor tissue of the cancer patient is reduced compared to the tumor tissue of a patient responsive to anti-PD-1 antibodies.

26. The therapeutic and / or prophylactic agent according to claim 1, wherein the liposome further comprises a phospholipid.

27. The therapeutic and / or prophylactic agent according to claim 26, wherein the phospholipid is one selected from the group consisting of phosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, soy lecithin, egg yolk lecithin, hydrogenated egg yolk lecithin, and hydrogenated soy lecithin, or a combination of two or more thereof.

28. The therapeutic and / or prophylactic agent according to claim 1, wherein the liposomes further comprise sterols.

29. The therapeutic and / or prophylactic agent according to claim 1, wherein the sterols are cholesterol.

30. The therapeutic and / or prophylactic agent according to claim 1, wherein the liposome further comprises polymer-modified lipids.

31. The therapeutic and / or prophylactic agent according to claim 30, wherein the polymer portion of the polymer-modified lipid is polyethylene glycol, polypropylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, methoxypolyethylene glycol, methoxypolypropylene glycol, methoxypolyvinyl alcohol, methoxypolyvinylpyrrolidone, ethoxypolyethylene glycol, ethoxypolypropylene glycol, ethoxypolyvinyl alcohol, ethoxypolyvinylpyrrolidone, propoxypolyethylene glycol, propoxypolypropylene glycol, propoxypolyvinyl alcohol, or propoxypolyvinylpyrrolidone.

32. The therapeutic and / or prophylactic agent according to claim 30, wherein the lipid portion of the polymer-modified lipid is phosphatidylethanolamine or diacylglycerol.

33. The liposomes containing the CHK1 inhibitor, (1) 40-70 mol% phospholipids, (2) 30-50 mol% cholesterol, and (3) 1 to 10 mol% polymer-modified lipids, A therapeutic and / or prophylactic agent according to claim 1, comprising:

34. The therapeutic and / or prophylactic agent according to claim 1, wherein the liposomes further contain an additive selected from the group consisting of inorganic acids, inorganic acid salts, organic acids, organic acid salts, sugars, buffers, antioxidants, and polymers.

35. The CHK1 inhibitor is defined by formula (1): 【Chemistry 1】 [In the formula, R １ C is a hydrogen atom, which may be substituted. １－６ Alkyl, optionally substituted C ３－１０ Cycloalkyl groups, optionally substituted 3- to 10-membered saturated heterocyclic groups, optionally substituted C ６－１０ Represents an aryl group, or a 5- to 12-membered heteroaryl group which may be substituted. R ２ is a hydrogen atom, a halogen atom, cyano, nitro, carboxyl, sulfonic acid, phosphoric acid, -OR ３ , -SR ３ , -COR ４ , -CO ２ R ４ , -CONR ５ R ６ , -SO ２ R ４ , -SO ２ NR ５ R ６ , -O COR ４ , -OCO ２ R ４ , -OCONR ５ R ６ , -NR ５ R ６ , -NR ７ COR ４ , -NR ７ CO ２ R ４ , -NR ７ CONR ５ R ６ , -NR ７ SO ２ R ４ , -NR ７ SO ２ NR ５ R ６ , optionally substituted C １－６ alkyl, optionally substituted C ２－６ alkenyl, optionally substituted C ２－６ alkynyl, optionally substituted C ３－１０ cycloalkyl, optionally substituted 3- to 10-membered saturated heterocyclic group, optionally substituted C ６－１０ aryl, or optionally substituted 5- to 12-membered heteroaryl, and R ３ is a hydrogen atom or C １－６ Represents alkyl, R ４ C １－６ Represents alkyl, R ５ , R ６ and R ７ Each of these is independently a hydrogen atom, or C １－６ R represents alkyl, and here it is bonded to the same nitrogen atom. ５ and R ６ Both are C １－６ In the case of alkyl groups, these may form a 3- to 8-membered nitrogen-containing saturated heterocycle together with the nitrogen atom to which they are bonded. X, Y, and Z are each independent of CR ８ or represent a nitrogen atom, where X, Y and Z are simultaneously CR ８ Instead, R ８ When there are a plurality of them, each independently represents a hydrogen atom, a halogen atom, cyano, nitro, carboxyl, sulfonic acid, phosphoric acid, -OR ９ -SR ９ -COR １０ -CO ２ R １０ -CONR １１ R １２ -SO ２ R １０ -SO ２ NR １１ R １２ -O COR １０ -OCO ２ R １０ -OCONR １１ R １２ -NR １１ R １２ -NR １３ COR １０ -NR １３ CO ２ R １０ -NR １３ CONR １１ R １２ -NR １３ SO ２ R １０ -NR １３ SO ２ NR １１ R １２ represents an optionally substituted C １－６ alkyl, an optionally substituted C ２－６ alkenyl, an optionally substituted C ２－６ alkynyl, an optionally substituted C ３－１０ cycloalkyl, an optionally substituted 3- to 10-membered saturated heterocyclic group, an optionally substituted C ６－１０ aryl, or an optionally substituted 5- to 12-membered heteroaryl, R ９ is a hydrogen atom or C １－６ Represents alkyl, R １０ C １－６ Represents alkyl, R １１ , R １２ and R １３ Each of these is independently a hydrogen atom or C １－６ R represents alkyl, and here it is bonded to the same nitrogen atom. １１ and R １２ Both are C １－６ In the case of alkyl groups, these may form a 3- to 8-membered nitrogen-containing saturated heterocycle together with the nitrogen atom to which they are bonded. L may be a single bond or a substituted C. １－６ Represents alkylene, V is a single bond, and C may be substituted. ３－１０ Represents a cycloalkylene or a 3- to 10-membered divalent saturated heterocyclic group which may be substituted, W may be a single bond or a substituted C. １－６ Represents alkylene, Q is a hydrogen atom or NHR １４ This represents, R １４ C is a hydrogen atom, which may be substituted. １－６ Alkyl, optionally substituted C ３－１０ Represented by [a cycloalkyl group or a substituted 3- to 10-membered saturated heterocyclic group] The therapeutic and / or prophylactic agent according to claim 1, which is a compound or a pharmaceutically acceptable salt thereof.

36. Equation (1) becomes the following equation (2): 【Chemistry 2】 [In the formula, X, Y, and Z are each independent of CR ８ or represent a nitrogen atom, where X, Y and Z are simultaneously CR ８ Instead, R ８ If there are multiple, each one is independent. hydrogen atom, Fluorine atom, Chlorine atom, Bromine atom, C １－６ Alkyl (the alkyl group is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, -NR １６ R １７ (which may be substituted with one or two identical or different substituents selected from the group consisting of , and cyano), 5-6 member heteroaryl (the heteroaryl is composed of a fluorine atom, a hydroxyl group, and C １－３ Alkyl, C １－３ Alkoxy, -NR １６ R １７ (May be substituted with one or two identical or different substituents selected from the group consisting of , and cyano) L is Single bond, or C １－６ Alkylene (the alkylene contains a fluorine atom, a hydroxyl group, C １－３ Alkoxy, -NR １６ R １７ (May be substituted with one to three identical or different substituents selected from the group consisting of , and cyano) V is, single bond, C ３－１０ Cycloalkylene (the cycloalkylene contains a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, -NR １６ R １７ (These may be substituted with one to three identical or different substituents selected from the group consisting of , and cyano), or A 3-10 membered divalent saturated heterocyclic group (the saturated heterocyclic group may be substituted with a fluorine atom, a hydroxyl group, one or two hydroxyl groups, or a fluorine atom) C １－３ Alkyl, C １－３ Alkoxy, -NR １６ R １７ (May be substituted with one to three identical or different substituents selected from the group consisting of , and cyano) W is Single bond, or C １－６ Alkylene (the alkylene contains a fluorine atom, a hydroxyl group, C １－３ Alkoxy, -NR １６ R １７ (May be substituted with one to three identical or different substituents selected from the group consisting of , and cyano) Q is a hydrogen atom, or NHR １４ This represents, R １４ teeth, hydrogen atom, C １－６ Alkyl (the alkyl group is a fluorine atom, a hydroxyl group, C １－３ Alkoxy, -NR １６ R １７ (These may be substituted with one to three identical or different substituents selected from the group consisting of cyano, etc.) C ３－１０ Cycloalkyl (the cycloalkyl is a fluorine atom, a hydroxyl group, C １－３ Alkyl, C １－３ Alkoxy, -NR １６ R １７ (These may be substituted with one to three identical or different substituents selected from the group consisting of , and cyano), or A saturated heterocyclic group with 3 to 10 members (the saturated heterocyclic group contains a fluorine atom, a hydroxyl group, and C １－３ Alkyl, C １－３ Alkoxy, -NR １６ R １７ (May be substituted with one to three identical or different substituents selected from the group consisting of , and cyano) R １６ and R １７ Each of these is independently a hydrogen atom, or C １－６ R represents alkyl, １６ or R １７ If there are multiple, R １６ or R １７ Each of these may be the same or different, and here, R bonded to the same nitrogen atom １６ and R １７ Both are C １－６ In the case of alkyl groups, these may form a 3- to 8-membered nitrogen-containing saturated heterocycle together with the nitrogen atom to which they are bonded. The therapeutic and / or prophylactic agent according to claim 35.

37. Equation (1) becomes the following equation (3): 【Transformation 3】 [In the formula, R ８ａ and R ８ｂ Each of these is independently a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－３ Represents alkyl, L may be a single bond or C may be substituted with one hydroxyl group. １－６ Represents alkylene, V is, single bond, C ３－７ Cycloalkylene (the cycloalkylene has a hydroxyl group and C １－３ (May be substituted with one substituent selected from the group consisting of alkyl groups), or A 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group comprises a fluorine atom, a cyano group, a hydroxyl group, and C １－３ This represents a molecule that may be substituted with one substituent selected from the group consisting of alkyl groups. W is Single bond, or C may be substituted with one hydroxyl group. １－３ Represents alkylene, Q is a hydrogen atom or NH ２ It is expressed as] The therapeutic and / or prophylactic agent according to claim 35.

38. R ８ａ and R ８ｂ However, each is independently a hydrogen atom, a fluorine atom, or a chlorine atom. The therapeutic and / or preventive agent according to claim 37.

39. R ８ａ and R ８ｂ However, each is independently a hydrogen atom or a chlorine atom. The therapeutic and / or preventive agent according to claim 37.

40. L is C １－３ It is alkylene. The therapeutic and / or preventive agent according to claim 37.

41. V is a single bond. The therapeutic and / or preventive agent according to claim 37.

42. V is C ３－７ It is a cycloalkylene. The therapeutic and / or preventive agent according to claim 37.

43. W is a single bond. The therapeutic and / or preventive agent according to claim 37.

44. W is C １－３ It is alkylene. The therapeutic and / or preventive agent according to claim 37.

45. Q is NH ２ That is, The therapeutic and / or preventive agent according to claim 37.

46. Equation (1) becomes the following equation (4): 【Chemistry 4】 [In the formula, R ８ｂ and R ８ｃ Each of these is independently a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－３ Represents alkyl, L is Single bond, or C １－６ Represents alkylene (the alkylene may be substituted with one substituent selected from the group consisting of a fluorine atom, a hydroxyl group, and cyano), V is, single bond, C ３－７ Cycloalkylene (the cycloalkylene has a hydroxyl group and C １－３ (May be substituted with one substituent selected from the group consisting of alkyl groups), or A 3-7 member divalent saturated heterocyclic group (the saturated heterocyclic group may be substituted with a fluorine atom, a cyano group, a hydroxyl group, and 1-3 hydroxyl groups and a fluorine atom) C １－３ This represents a molecule that may be substituted with one substituent selected from the group consisting of alkyl groups. W is Single bond, or C may be substituted with one hydroxyl group. １－３ Represents alkylene, Q is a hydrogen atom, NH ２ [represented as , or NHMe] The therapeutic and / or prophylactic agent according to claim 35.

47. R ８ｂ and R ８ｃ However, it is a hydrogen atom. The therapeutic and / or prophylactic agent according to claim 46.

48. L, Single bond, or C may be substituted with one hydroxyl group or fluorine atom. １－６ It is alkylene. The therapeutic and / or prophylactic agent according to claim 46.

49. V is, single bond, C ３－７ Cycloalkylene, or A 3- to 7-membered divalent saturated heterocyclic group (the saturated heterocyclic group contains a fluorine atom, a hydroxyl group and C １－３ (It may be substituted with one substituent selected from the group consisting of alkyl groups.) The therapeutic and / or prophylactic agent according to claim 46.

50. W is Single bond, or C １－３ It is alkylene. The therapeutic and / or prophylactic agent according to claim 46.

51. Q is a hydrogen atom. The therapeutic and / or prophylactic agent according to claim 46.

52. Q is NH ２ That is, The therapeutic and / or prophylactic agent according to claim 46.

53. Q is NHMe. The therapeutic and / or prophylactic agent according to claim 46.

54. Equation (1) is given by the following equation (5): 【Transformation 5】 [In the formula, R ８ａ and R ８ｃ Each of these is independently a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, or C １－３ Represents alkyl, L may be a single bond or C may be substituted with one hydroxyl group. １－6 Represents alkylene, V is, single bond, C ３－７ Cycloalkylene (the cycloalkylene has a hydroxyl group and C １－３ (May be substituted with one substituent selected from the group consisting of alkyl groups), or A 3-7 member divalent saturated heterocyclic group (the saturated heterocyclic group may be substituted with a fluorine atom, a cyano group, a hydroxyl group, and one hydroxyl group) １－３ This represents a molecule that may be substituted with one substituent selected from the group consisting of alkyl groups. W is Single bond, or C may be substituted with one hydroxyl group. １－３ Represents alkylene, Q is a hydrogen atom, or NH ２ It is expressed as] The therapeutic and / or prophylactic agent according to claim 35.

55. R ８ａ and R ８ｃ However, it is a hydrogen atom. The therapeutic and / or prophylactic agent according to claim 54.

56. C may be substituted with one hydroxyl group. １－6 It is alkylene. The therapeutic and / or prophylactic agent according to claim 54.

57. V is, single bond, C ３－７ Cycloalkylene or A 3-7 member divalent saturated heterocyclic group (the saturated heterocyclic group may be substituted with a hydroxyl group and one hydroxyl group) C １－３ (It may be substituted with one substituent selected from the group consisting of alkyl groups.) The therapeutic and / or prophylactic agent according to claim 54.

58. W is Single bond, or C １－３ It is alkylene. The therapeutic and / or prophylactic agent according to claim 54.

59. Q is a hydrogen atom. The therapeutic and / or prophylactic agent according to claim 54.

60. Q is NH ２ That is, The therapeutic and / or prophylactic agent according to claim 54.

61. The therapeutic and / or prophylactic agent according to claim 35, comprising a compound selected from the following compounds or a pharmaceutically acceptable salt thereof: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridine-3-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[2-(3-aminopropoxy)-6-chloro-4-methoxypyridine-3-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[3-(3-aminopropoxy)-5-methoxypyridine-4-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[4-(3-aminopropoxy)-2-methoxypyridine-3-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile, 5-[(5-{3-[(3-fluoroazetidine-3-yl)methoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{2-methoxy-4-[(3-methylazetidine-3-yl)methoxy]pyridine-3-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{4-[(3-hydroxyazetidine-3-yl)methoxy]-2-methoxypyridine-3-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-{[5-(4-{[3-(hydroxymethyl)azetidine-3-yl]methoxy}-2-methoxypyridine-3-yl)-1H-pyrazole-3-yl]amino}pyrazine-2-carbonitrile, 5-[(5-{3-[(3R)-3-aminobutoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(3S)-3-aminobutoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-{[5-(3-{[1-(aminomethyl)cyclopropyl]methoxy}-5-methoxypyridine-4-yl)-1H-pyrazole-3-yl]amino}pyrazine-2-carbonilicate, 5-[(5-{3-methoxy-5-[(morpholine-2-yl)methoxy]pyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-methoxy-5-[(morpholine-2-yl)methoxy]pyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(2S)-3-amino-2-hydroxypropoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, N-{5-[2-(3-aminopropoxy)-4-methoxypyridine-3-yl]-1H-pyrazole-3-yl}-5-chloropyrazine-2-amine, N-{5-[2-(3-aminopropoxy)-4-methoxypyridine-3-yl]-1H-pyrazole-3-yl}-5-(trifluoromethyl)pyrazine-2-amine, 5-({5-[4-(3-aminopropoxy)-6-methoxypyrimidine-5-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[3-(azetidine-3-yl)methoxy-5-methoxypyridine-4-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile, 5-{[5-(3-{[(1R,3S)-3-aminocyclohexyl]oxy}-5-methoxypyridine-4-yl)-1H-pyrazole-3-yl]amino}pyrazine-2-carbonitrile, (S)-5-[(5-{3-[(3-fluoropyrrolidine-3-yl)methoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, (S)-5-[(5-{3-methoxy-[5-(pyrroridine-3-yl)methoxy]pyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, (R)-5-[(5-{3-[(3-fluoropyrrolidine-3-yl)methoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-{[5-(4-{[1-(aminomethyl)cyclopropyl]methoxy}-6-methoxypyrimidine--yl)-1H-pyrazole-3-yl]amino}pyrazine-2-carbonitrile, 5-({5-[3-(3-aminopropoxy)-5-(fluoromethoxy)pyridine-4-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile, 5-[(5-{3-methoxy-5-[(3-methylazetidine-3-yl)methoxy]pyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-{[5-(3-{[3-(difluoromethyl)azetidine-3-yl]methoxy}-5-methoxypyridine-4-yl)-1H-pyrazole-3-yl]amino}pyrazine-2-carbonilicate, 5-[(5-{3-[(2S)-3-amino-2-methylpropoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(2R)-3-amino-2-methylpropoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(2S)-3-amino-2-fluoropropoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(2R)-3-amino-2-fluoropropoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-methoxy-5-[3-(methylamino)propoxy]pyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-{[5-(3-{[(1R,3R)-3-aminocyclopentyl]oxy}-5-methoxypyridine-4-yl)-1H-pyrazole-3-yl]amino}pyrazine-2-carbonitrile, 5-[(5-{3-[(1R)-1-(azetidine-3-yl)ethoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-[(5-{3-[(1R)-1-(3-hydroxyazetidine-3-yl)ethoxy]-5-methoxypyridine-4-yl}-1H-pyrazole-3-yl)amino]pyrazine-2-carbonitrile, 5-{[5-(3-methoxy-5-{[(1R,3R)-3-(methylamino)cyclopentyl]oxy}pyridine-4-yl)-1H-pyrazole-3-yl]amino}pyrazine-2-carbonitrile, 5-{[5-(3-{[(1R,2S,4S,5S)-4-aminobicyclo[3.1.0]hexane-2-yl]oxy}-5-methoxypyridine-4-yl)-1H-pyrazole-3-yl]amino}pyrazine-2-carbonitrile, 5-{[5-(2-{[1-(aminomethyl)cyclopropyl]methoxy}-4-methoxypyridine-3-yl)-1H-pyrazole-3-yl]amino}pyrazine-2-carbonilicate, 5-{[5-(4-{[1-(aminomethyl)cyclopropyl]methoxy}-2-methoxypyridine-3-yl)-1H-pyrazole-3-yl]amino}pyrazine-2-carbonilicate.

62. The therapeutic and / or prophylactic agent according to claim 35, comprising a compound selected from the following compounds or a pharmaceutically acceptable salt thereof: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridine-3-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[3-(3-aminopropoxy)-5-methoxypyridine-4-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile, 5-({5-[4-(3-aminopropoxy)-2-methoxypyridine-3-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile.

63. The therapeutic and / or prophylactic agent according to claim 35, comprising a compound selected from the following compounds or a pharmaceutically acceptable salt thereof: 5-({5-[4-(3-aminopropoxy)-2-methoxypyridine-3-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile.

64. The therapeutic and / or prophylactic agent according to claim 35, comprising the following compounds or pharmaceutically acceptable salts thereof: 5-({5-[2-(3-aminopropoxy)-4-methoxypyridine-3-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile.

65. The therapeutic and / or prophylactic agent according to claim 35, comprising the following compounds or pharmaceutically acceptable salts thereof: 5-({5-[3-(3-aminopropoxy)-5-methoxypyridine-4-yl]-1H-pyrazole-3-yl}amino)pyrazine-2-carbonitrile.

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