Pharmaceutical composition for treating tumors

Abstract

The present invention provides a pharmaceutical composition for treating tumors, which comprises (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof and is administered in combination with an RAS inhibitor.

Description

Pharmaceutical composition for tumor treatment

[0001] The present invention relates to a pharmaceutical composition for treating tumors.

[0002] Ras proteins belong to a family of related proteins present in all eukaryotes, from yeast to humans (Non-Patent Document 1). It is believed that amino acid substitutions due to mutations in the RAS gene increase the proportion of activated RAS due to reduced GTPase function and reduced affinity for GAP. The resulting excessive signal transduction is believed to lead to carcinogenesis and accelerated cancer proliferation. There are four major Ras proteins: KRAS (KRAS4A and KRAS4B), NRAS, and HRAS, which are produced by three genes: KRAS, NRAS, and HRAS. KRAS is the most commonly mutated member of the RAS family and is considered the most common oncogenic gene driver in human cancer. KRAS mutations are commonly observed in non-small cell lung cancer, colon cancer, and pancreatic cancer (Non-Patent Document 2).

[0003] On the other hand, the Wnt / β-catenin signaling pathway is a signaling pathway that has been highly conserved throughout the evolution of animals and regulates gene expression involved in cell proliferation and differentiation, body axis and organ formation, etc. Abnormal activation of the Wnt / β-catenin signaling pathway is known to be observed in various cancers, including colorectal cancer and hepatocellular carcinoma (Non-Patent Document 3).

[0004] The IUPAC name of the compound represented by formula (I) is (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide. The compound represented by formula (I) is also called (6S,9aS)-N-benzyl-8-((6-(3-(4-ethylpiperazin-1-yl)azetidin-1-yl)pyridin-2-yl)methyl)-6-((2-fluoro-4-hydroxyphenyl)methyl)-4,7-dioxo-2-(prop-2-en-1-yl)-octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide, and is known as a compound having a Wnt pathway modulating activity (Patent Documents 1 and 2).

[0005] International Publication No. WO 2015 / 098853 International Publication No. WO 2016 / 208576

[0006] Santos et al., Structural and functional properties of ras proteins, FASEB J. 1989, 3(10),2151-2163Interaction between Wnt / β-catenin and RAS-ERK pathways and an anti-cancer strategy via degradations of β-catenin and RAS by targeting the Wnt / β-catenin pathway. npj Precision Onc 2, 5 (2018)Fodde, et al., Wnt / β-catenin signaling in cancer stemness and malignant behavior, Current Opinion in Cell Biology 2007, 19:150-158

[0007] In recent years, active development of drugs targeting RAS has been underway. For example, as a molecular targeted drug for KRAS mutations, sotorasib, a KRAS G12C inhibitor, is clinically used as a therapeutic agent for non-small cell lung cancer harboring KRAS G12 mutations. However, to date, no combination therapy using a RAS inhibitor and a Wnt / β-catenin pathway inhibitor has been reported.

[0008] An object of the present invention is to provide a novel pharmaceutical composition for tumor treatment.

[0009] As a result of extensive investigations, the present inventors have found that combined administration of (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide (hereinafter also referred to as the compound represented by formula (I)) or a pharmaceutically acceptable salt thereof and a RAS inhibitor exhibits a high antitumor effect, and have completed the present invention.

[0010] That is, the present disclosure is as follows: [1] A pharmaceutical composition for tumor treatment comprising (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is administered in combination with a RAS inhibitor. [2] The pharmaceutical composition according to [1], wherein (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof is administered simultaneously or at different times with a RAS inhibitor. [3] (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof The pharmaceutical composition according to [1] or [2], wherein the RAS inhibitor is N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide. [4] The pharmaceutical composition according to any one of [1] to [3], wherein the RAS inhibitor is a KRAS inhibitor. [5] The pharmaceutical composition according to [4], wherein the KRAS inhibitor is at least one selected from sotorasib (AMG 510), adagrasib (MRTX849), MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, mRNA-5671, JAB-21822, BI 1823911, MK-1084, ELI-002, SDGR5, and JAB-22000. [6] The pharmaceutical composition according to [4], wherein the KRAS inhibitor is sotorasib, adagrasib, or BI 1701963. [7] The pharmaceutical composition according to [4], wherein the KRAS inhibitor is a KRAS G12C inhibitor.[8] The pharmaceutical composition according to [7], wherein the KRAS G12C inhibitor is sotorasib or adagrasib. [9] The pharmaceutical composition according to any one of [1] to [8], wherein the tumor is at least one selected from non-small cell lung cancer, colon cancer, pancreatic cancer, mesothelioma, uterine cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, testicular cancer, small intestine cancer, appendix cancer, and neuroendocrine tumor.

[10] The pharmaceutical composition according to any one of [1] to [8], wherein the tumor is at least one selected from non-small cell lung cancer, colon cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[11] The pharmaceutical composition according to any one of [1] to [8], wherein the tumor is at least one selected from non-small cell lung cancer and colon cancer.

[12] A pharmaceutical composition for tumor treatment containing a RAS inhibitor, which is administered in combination with (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof.

[13] The pharmaceutical composition according to

[12] , wherein the RAS inhibitor is administered simultaneously or at different times with (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof.

[14] (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof ,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide.

[15] The pharmaceutical composition according to any one of

[12] to

[14] , wherein the RAS inhibitor is a KRAS inhibitor.

[16] The pharmaceutical composition according to

[15] , wherein the KRAS inhibitor is at least one selected from sotorasib, adagrasib, MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, mRNA-5671, JAB-21822, BI 1823911, MK-1084, ELI-002, SDGR5, and JAB-22000.

[17] The pharmaceutical composition according to

[15] , wherein the KRAS inhibitor is sotorasib, adagrasib, or BI 1701963.

[18] The pharmaceutical composition according to

[15] , wherein the KRAS inhibitor is a KRAS G12C inhibitor.

[19] The pharmaceutical composition according to

[18] , wherein the KRAS G12C inhibitor is sotorasib or adagrasib.

[20] The pharmaceutical composition according to any one of

[12] to

[19] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, mesothelioma, uterine cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, testicular cancer, small intestine cancer, appendix cancer, and neuroendocrine tumor.

[21] The pharmaceutical composition according to any one of

[12] to

[19] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[22] The pharmaceutical composition according to any one of

[12] to

[19] , wherein the tumor is at least one selected from non-small cell lung cancer and colorectal cancer.

[23] A tumor therapeutic agent comprising (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof, which is administered in combination with a RAS inhibitor.

[24] The therapeutic agent according to

[23] , wherein (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof is administered simultaneously or at different times with a RAS inhibitor.

[25] (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof The therapeutic agent according to

[23] or

[24] , wherein the RAS inhibitor is N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide.

[26] The therapeutic agent according to any one of

[23] to

[25] , wherein the RAS inhibitor is a KRAS inhibitor.

[27] The therapeutic agent according to

[26] , wherein the KRAS inhibitor is at least one selected from sotorasib, adagrasib, MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, mRNA-5671, JAB-21822, BI 1823911, MK-1084, ELI-002, SDGR5, and JAB-22000.

[28] The therapeutic agent according to

[26] , wherein the KRAS inhibitor is sotorasib, adagrasib, or BI 1701963.

[29] The therapeutic agent according to

[26] , wherein the KRAS inhibitor is a KRAS G12C inhibitor.

[30] The therapeutic agent according to

[29] , wherein the KRAS G12C inhibitor is sotorasib or adagrasib.

[31] The therapeutic agent according to any one of

[23] to

[30] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, mesothelioma, uterine cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, testicular cancer, small intestine cancer, appendix cancer, and neuroendocrine tumor.

[32] The therapeutic agent according to any one of

[23] to

[30] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[33] The therapeutic agent according to any one of

[23] to

[30] , wherein the tumor is at least one selected from non-small cell lung cancer and colorectal cancer.

[34] A tumor therapeutic agent containing a RAS inhibitor, which is administered in combination with (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof.

[35] The therapeutic agent according to

[34] , wherein the RAS inhibitor is administered simultaneously or at different times with (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof.

[36] (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof The therapeutic agent according to

[34] or

[35] , wherein the RAS inhibitor is N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide.

[37] The therapeutic agent according to any of

[34] to

[36] , wherein the RAS inhibitor is a KRAS inhibitor.

[38] The therapeutic agent according to

[37] , wherein the KRAS inhibitor is at least one selected from sotorasib, adagrasib, MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, mRNA-5671, JAB-21822, BI 1823911, MK-1084, ELI-002, SDGR5, and JAB-22000.

[39] The therapeutic agent according to

[37] , wherein the KRAS inhibitor is sotorasib, adagrasib, or BI 1701963.

[40] The therapeutic agent according to

[37] , wherein the KRAS inhibitor is a KRAS G12C inhibitor.

[41] The therapeutic agent according to

[40] , wherein the KRAS G12C inhibitor is sotorasib or adagrasib.

[42] The therapeutic agent according to any one of

[34] to

[41] , wherein the tumor is at least one selected from non-small cell lung cancer, colon cancer, pancreatic cancer, mesothelioma, uterine cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, testicular cancer, small intestine cancer, appendix cancer, and neuroendocrine tumor.

[43] The therapeutic agent according to any one of

[34] to

[41] , wherein the tumor is at least one selected from non-small cell lung cancer, colon cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[44] The therapeutic agent according to any one of

[34] to

[41] , wherein the tumor is at least one selected from non-small cell lung cancer and colon cancer.

[45] A method for treating a tumor, comprising administering (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof, and a RAS inhibitor to a patient in need thereof.

[46] The method according to

[45] , wherein (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof and a RAS inhibitor are administered simultaneously or at different times.

[47] (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof

[48] ​​The method according to any one of

[45] to

[47] , wherein the RAS inhibitor is a KRAS inhibitor.

[49] The method according to

[48] , wherein the KRAS inhibitor is at least one selected from sotorasib (AMG 510), adagrasib (MRTX849), MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, mRNA-5671, JAB-21822, BI 1823911, MK-1084, ELI-002, SDGR5, and JAB-22000.

[50] The method according to

[48] , wherein the KRAS inhibitor is sotorasib, adagrasib, or BI 1701963.

[51] The method according to

[48] , wherein the KRAS inhibitor is a KRAS G12C inhibitor.

[52] The method of

[51] , wherein the KRAS G12C inhibitor is sotorasib or adagrasib.

[53] The method of any of

[45] to

[52] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, mesothelioma, uterine cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, testicular cancer, small intestine cancer, appendix cancer, and neuroendocrine tumor.

[54] The method of any of

[45] to

[52] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[55] The method of any of

[45] to

[52] , wherein the tumor is at least one selected from non-small cell lung cancer and colorectal cancer.

[56] Use of (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof for the manufacture of a pharmaceutical composition for tumor treatment to be administered in combination with a RAS inhibitor.

[57] The use according to

[56] , wherein a pharmaceutical composition for tumor treatment containing (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof is administered simultaneously or at different times with a RAS inhibitor.

[58] (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof The use according to

[56] or

[57] , wherein the RAS inhibitor is N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide.

[59] The use according to any one of

[56] to

[58] , wherein the RAS inhibitor is a KRAS inhibitor.

[60] The use according to

[59] , wherein the KRAS inhibitor is at least one selected from sotorasib, adagrasib, MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, mRNA-5671, JAB-21822, BI 1823911, MK-1084, ELI-002, SDGR5, and JAB-22000.

[61] The use according to

[59] , wherein the KRAS inhibitor is sotorasib, adagrasib, or BI 1701963.

[62] The use according to

[59] , wherein the KRAS inhibitor is a KRAS G12C inhibitor.

[63] The use according to

[62] , wherein the KRAS G12C inhibitor is sotorasib or adagrasib.

[64] The use according to any one of

[56] to

[63] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, mesothelioma, uterine cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, testicular cancer, small intestine cancer, appendix cancer, and neuroendocrine tumor.

[65] The use according to any one of

[56] to

[63] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[66] The use according to any one of

[56] to

[63] , wherein the tumor is at least one selected from non-small cell lung cancer and colorectal cancer.

[67] Use of a RAS inhibitor for the manufacture of a pharmaceutical composition for tumor treatment, to be administered in combination with (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof.

[68] The use according to

[67] , wherein the RAS inhibitor is administered simultaneously or at different times with (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof.

[69] (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof The use according to

[67] or

[68] , wherein the RAS inhibitor is N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide.

[70] The use according to any one of

[67] to

[69] , wherein the RAS inhibitor is a KRAS inhibitor.

[71] The use according to

[70] , wherein the KRAS inhibitor is at least one selected from sotorasib, adagrasib, MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, mRNA-5671, JAB-21822, BI 1823911, MK-1084, ELI-002, SDGR5, and JAB-22000.

[72] The use according to

[70] , wherein the KRAS inhibitor is sotorasib, adagrasib, or BI 1701963.

[73] The use according to

[70] , wherein the KRAS inhibitor is a KRAS G12C inhibitor.

[74] The use according to

[73] , wherein the KRAS G12C inhibitor is sotorasib or adagrasib.

[75] The use according to any one of

[67] to

[74] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, mesothelioma, uterine cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, testicular cancer, small intestine cancer, appendix cancer, and neuroendocrine tumor.

[76] The use according to any one of

[67] to

[74] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[77] The use according to any one of

[67] to

[74] , wherein the tumor is at least one selected from non-small cell lung cancer and colorectal cancer.

[78] (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof for use in tumor treatment, administered in combination with a RAS inhibitor.

[79] (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof is administered simultaneously or at different times with a RAS inhibitor. (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof according to

[78] , which is administered to (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof is (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)- The (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide according to

[78] or

[79] , which is 4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide, or a pharmaceutically acceptable salt thereof.

[81] The (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof according to any one of

[78] to

[80] , wherein the RAS inhibitor is a KRAS inhibitor.

[82] KRAS inhibitors include sotorasib, adagrasib, MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, mRNA-5671, JAB-21822, and BI 1823911, MK-1084, ELI-002, SDGR5, and JAB-22000. The (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof according to

[81] , which is at least one selected from the group consisting of 1823911, MK-1084, ELI-002, SDGR5, and JAB-22000.

[83] The (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof according to

[81] , wherein the KRAS inhibitor is sotorasib, adagrasib, or BI 1701963.

[84] (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof according to

[81] , wherein the KRAS inhibitor is a KRAS G12C inhibitor.

[85] The (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof according to

[84] , wherein the KRAS G12C inhibitor is sotorasib or adagrasib.

[86] The (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof according to any of

[78] to

[85] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, mesothelioma, uterine cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, testicular cancer, small intestine cancer, appendix cancer, and neuroendocrine tumor.

[87] The (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof according to any one of

[78] to

[85] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[88] The (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof according to any one of

[78] to

[85] , wherein the tumor is at least one selected from non-small cell lung cancer and colorectal cancer.

[89] A RAS inhibitor for use in tumor treatment, administered in combination with (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof:

[90] The RAS inhibitor according to

[89] , wherein the RAS inhibitor is administered simultaneously or at different times with (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof.

[91] (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof The RAS inhibitor according to

[89] or

[90] , wherein the RAS inhibitor is N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide.

[92] The RAS inhibitor according to any one of

[89] to

[91] , wherein the RAS inhibitor is a KRAS inhibitor.

[93] The RAS inhibitor according to

[92] , wherein the KRAS inhibitor is at least one selected from sotorasib, adagrasib, MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, mRNA-5671, JAB-21822, BI 1823911, MK-1084, ELI-002, SDGR5, and JAB-22000.

[94] The RAS inhibitor according to

[92] , wherein the KRAS inhibitor is sotorasib, adagrasib, or BI 1701963.

[95] The RAS inhibitor according to

[92] , wherein the KRAS inhibitor is a KRAS G12C inhibitor.

[96] The RAS inhibitor according to

[95] , wherein the KRAS G12C inhibitor is sotorasib or adagrasib.

[97] The RAS inhibitor according to any one of

[89] to

[96] , wherein the tumor is at least one selected from non-small cell lung cancer, colon cancer, pancreatic cancer, mesothelioma, uterine cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, testicular cancer, small intestine cancer, appendix cancer, and neuroendocrine tumor.

[98] The RAS inhibitor according to any one of

[89] to

[96] , wherein the tumor is at least one selected from non-small cell lung cancer, colon cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

[99] The RAS inhibitor according to any one of

[89] to

[96] , wherein the tumor is at least one selected from non-small cell lung cancer and colon cancer.

[100] The pharmaceutical composition, therapeutic agent, or use according to any one of [1] to

[11] ,

[23] to

[33] , and

[56] to

[66] , wherein the pharmaceutical composition or therapeutic agent containing (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof is for oral administration.

[101] The pharmaceutical composition, therapeutic agent, therapeutic method, use, compound for use or a pharmaceutically acceptable salt thereof, or RAS inhibitor for use according to any one of

[12] to

[22] ,

[34] to

[55] , and

[67] to

[99] , wherein (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof is orally administered.

[102] The pharmaceutical composition, therapeutic agent, therapeutic method, use, compound for use or a pharmaceutically acceptable salt thereof, or RAS inhibitor for use according to any one of [1] to [8],

[12] to

[19] ,

[23] to

[30] ,

[34] to

[41] ,

[45] to

[52] ,

[56] to

[63] ,

[67] to

[74] ,

[78] to

[85] , and

[89] to

[96] , wherein the tumor is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, uterine cancer, bile duct cancer, and bladder cancer.

[0011] The combined administration of (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof and a RAS inhibitor exhibits a high antitumor effect.

[0012] 1 is a graph showing the antitumor effect of combined administration of the compound represented by formula (I) (abbreviated as E7386 in the figure) and a KRAS inhibitor in KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358. 2 is a graph showing the antitumor effect of combined administration of the compound represented by formula (I) (abbreviated as E7386 in the figure) and a KRAS inhibitor in a transplant model of KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358. 3 is a graph showing the cell growth inhibitory activity of combined treatment of the compound represented by formula (I) (abbreviated as E7386 in the figure) and a KRAS inhibitor in KRAS G12C mutation-positive human pancreatic cancer cell line MIA PaCa-2.

[0023] Figure 1 shows the cell growth inhibitory activity of a compound represented by formula (I) (abbreviated as E7386 in the figure) and a KRAS inhibitor in a KRAS G12C mutation-positive human bladder cancer cell line UM-UC-3.

[0024] Figure 2 shows the cell growth inhibitory activity of a compound represented by formula (I) (abbreviated as E7386 in the figure) and a KRAS inhibitor in a KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358.

[0025] Figure 3 shows the cell growth inhibitory activity of a compound represented by formula (I) (abbreviated as E7386 in the figure) and a KRAS inhibitor in a KRAS G12D mutation-positive human cholangiocarcinoma cell line SNU-869.

[0023] Figure 1 shows the cell growth inhibitory activity of combined treatment of the compound represented by formula (I) (abbreviated as E7386 in the figure) with a KRAS inhibitor in a KRAS G12D mutation-positive human uterine cancer cell line HEC-1-B.

[0024] Figure 2 shows the cell growth inhibitory activity of combined treatment of the compound represented by formula (I) (abbreviated as E7386 in the figure) with a KRAS inhibitor in a KRAS G12C mutation-positive human pancreatic cancer cell line MIA PaCa-2.

[0025] Figure 3 shows the antitumor effect of combined administration of the compound represented by formula (I) (abbreviated as E7386 in the figure) with a KRAS inhibitor in a transplant model of a KRAS G12C mutation-positive human colon cancer cell line SW837. FIG. 1 shows the antitumor effect of combined administration of the compound represented by formula (I) (abbreviated as E7386 in the figure) and a KRAS inhibitor in a transplant model of the KRAS G12C mutation-positive human pancreatic cancer cell line MIA PaCa-2.1 is a graph showing the antitumor effect of combined administration of the compound represented by formula (I) (abbreviated as E7386 in the figure) and a KRAS inhibitor in a transplant model of KRAS G12C mutation-positive human bladder cancer cell line UM-UC-3. 2 is a graph showing the antitumor effect of combined administration of the compound represented by formula (I) (abbreviated as E7386 in the figure) and a KRAS inhibitor in a transplant model of KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358.

[0013] The following describes embodiments of the present disclosure. The following embodiments are examples for explaining the present disclosure, and are not intended to limit the present disclosure to these embodiments. The present disclosure can be implemented in various forms without departing from the spirit and scope of the present disclosure.

[0014] In the present disclosure, the term "pharmaceutically acceptable salt" is not particularly limited as long as it forms a salt with the compound represented by formula (I), and examples thereof include inorganic acid salts, organic acid salts, inorganic base salts, organic base salts, acidic or basic amino acid salts, etc.

[0015] Examples of inorganic acid salts include hydrochlorides, hydrobromides, sulfates, nitrates, and phosphates. Examples of organic acid salts include carboxylates such as acetates, succinates, fumarates, maleates, tartrates, citrates, lactates, stearates, benzoates, and mandelates, and sulfonates such as methanesulfonates, ethanesulfonates, p-toluenesulfonates, and benzenesulfonates.

[0016] Examples of inorganic base salts include alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts and magnesium salts, aluminum salts, and ammonium salts. Examples of organic base salts include diethylamine salts, diethanolamine salts, meglumine salts, and N,N'-dibenzylethylenediamine salts.

[0017] Examples of acidic amino acid salts include aspartate and glutamate, and examples of basic amino acid salts include arginine salt, lysine salt, and ornithine salt.

[0018] The compound represented by formula (I) is (6S,9aS)-N-benzyl-8-((6-(3-(4-ethylpiperazin-1-yl)azetidin-1-yl)pyridin-2-yl)methyl)-6-((2-fluoro-4-hydroxyphenyl)methyl)-4,7-dioxo-2-(prop-2-en-1-yl)-octahydro-1H-pyrazino[2,1-c][1,2,4]triazine-1-carboxamide Its IUPAC name is (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide. In this specification, the compound represented by formula (I) is also referred to as "E7386."

[0019] The compound represented by formula (I) can be prepared by the method described in US Pat. No. 9,174,998 or US Pat. No. 1,025,9817, and has pharmacological activity including antitumor activity.

[0020] In the present disclosure, one embodiment of the "compound represented by formula (I) or a pharmaceutically acceptable salt thereof" is, for example, the compound represented by formula (I), i.e., (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide.

[0021] A "pharmaceutical composition" or "therapeutic agent" in this disclosure is understood to mean a substance that produces a desired effect in a tissue, animal, mammal, human, or other subject.

[0022] In this disclosure, "treating" and its derivatives refer to therapeutic therapy. With reference to a specific condition, treating means (1) ameliorating the condition or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade leading to or causing the condition, or (b) one or more biological manifestations of the condition, (3) alleviating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0023] In this disclosure, a "therapeutically effective amount" refers to an amount that is effective to elicit a desired biological or pharmaceutical response in a tissue system, animal, or human. In one embodiment, a therapeutically effective amount refers to an amount effective, at dosages and for periods of time necessary, to achieve a desired therapeutic result (e.g., amelioration or prevention of symptoms, or prolongation of survival). In one embodiment, a therapeutically effective amount is an amount that does not exceed the maximum tolerated dose.

[0024] In the present disclosure, the term "patient" refers to a mammal, particularly a human, suffering from or suspected of suffering from a tumor. Mammals include, for example, guinea pigs, mice, rats, gerbils, cats, rabbits, dogs, cows, goats, sheep, horses, monkeys, chimpanzees, and humans. In some embodiments, the patient is a human. The pharmaceutical compositions, therapeutic agents, or treatment methods of the present disclosure are particularly useful for treating human patients with tumors.

[0025] The terms "a," "an," "the," and similar terms as used in the context of this disclosure (especially in the context of the claims) are intended to encompass both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0026] The "pharmaceutical composition" or "therapeutic agent" of the present disclosure can be administered by injection (intravenous injection, intra-arterial injection, local injection), oral, nasal, transdermal, pulmonary, or ophthalmic administration. Examples of injections include intravenous, subcutaneous, intradermal, and intra-arterial injections, as well as local injections into target cells or organs. Examples of dosage forms of the pharmaceutical composition or therapeutic agent for oral administration include tablets, powders, granules, syrups, capsules, and oral liquids. Examples of dosage forms of the pharmaceutical composition for parenteral administration include injections, drip infusions, eye drops, ointments, suppositories, suspensions, poultices, lotions, aerosols, and plasters. In one embodiment, the pharmaceutical composition is an injection or drip infusion. The pharmaceutical composition of the present disclosure can be formulated, for example, by a method described in the 18th Edition of the Japanese Pharmacopoeia (JP), the United States Pharmacopoeia (USP), or the European Pharmacopoeia (EP).

[0027] In one embodiment of the present disclosure, the compound of formula (I) or a pharmaceutically acceptable salt thereof is orally administered to a patient. In another embodiment, the pharmaceutical composition or therapeutic agent containing the compound of formula (I) or a pharmaceutically acceptable salt thereof is for oral administration.

[0028] In one embodiment, the pharmaceutical composition or therapeutic agent according to the present disclosure can further comprise a pharmaceutically acceptable carrier.

[0029] Pharmaceutically acceptable carriers include, for example, liquid or solid fillers, diluents, excipients, manufacturing aids, solvent encapsulating materials, and the like.

[0030] In the present disclosure, two subjects administered in combination can be administered simultaneously or at different times. The term "administered simultaneously" in "administered simultaneously or at different times" refers to two subjects administered in combination at the same time or substantially at the same time via the same administration route (simultaneously administration), and to two subjects administered simultaneously or substantially at the same time via different administration routes (separately administration). Here, "administered simultaneously" also includes cases where two subjects are administered as a single formulation. Furthermore, in the present disclosure, the term "administered at different times" in "administered simultaneously or at different times" refers to two subjects administered in combination at different times via the same administration route or different administration routes (sequentially administration). More specifically, this refers to an administration method in which administration of one of the two subjects is completed before administration of the other is initiated. "Administered at different times" also includes cases where the administration schedules for the two subjects differ in administration frequency or duration.

[0031] The dosage of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof when administered alone varies significantly depending on the type of disease, the patient's age, sex, body weight, and severity of symptoms. The dosage of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is not particularly limited. Typically, when orally administered to an adult (body weight 60 kg) or child, the dosage is 0.1 to 5000 mg, 0.5 to 3000 mg, or 1.0 to 1000 mg of the compound represented by formula (I) per day, and in one embodiment, 100 mg to 300 mg. This dosage can typically be administered once per day or over multiple days, or in two to six divided doses per day. The dosage and administration schedule can be modified when one or more additional chemotherapeutic agents are used. The administration schedule can be determined by the physician treating a particular patient. In one embodiment, for example, the compound represented by formula (I) is orally administered twice per day to a human subject at a dosage of 80 mg to 120 mg per dose. The specific dose, administration route, administration frequency, administration cycle, etc. of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof are appropriately determined taking into consideration the type of target disease, the age, sex, weight, and severity of symptoms of the patient, other drugs to be administered in combination, etc.

[0032] RAS inhibitors in the present disclosure may include any compound or biomolecule that (a) binds to and inhibits the function of a biomolecule (e.g., a protein or nucleic acid) having RAS, (b) binds to and degrades a biomolecule (e.g., a protein or nucleic acid) having RAS, (c) suppresses the expression of a biomolecule (e.g., a protein or nucleic acid) having RAS, (d) inhibits the GDP-GTP exchange reaction of RAS by inhibiting the protein-protein interaction between RAS and a guanine nucleotide exchange factor (e.g., SOS1, SOS2), or (e) functions as a vaccine to stimulate immunity against mutant RAS.

[0033] In the present disclosure, examples of RAS inhibitors include sotorasib (AMG 510), adagrasib (MRTX849), MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, D-1553, mRNA5671, BI 2852, SCH-54292, TLN-4601, Salirasib, Deltarasin, JAB-21822, and BI 1823911, MK-1084, ELI-002, SDGR5, JAB-22000, and ASP3082.

[0034] Sotorasib (also known as AMG 510) has the following structure: It refers to the compound having the chemical name 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one. The structural formula and preparation of sotorasib (AMG 510) are described in U.S. Pat. No. 10,519,146, from Column 411, line 10 to Column 415, line 19, which are incorporated herein by reference.

[0035] Adaglasib (also known as MRTX849) has the following structure: It refers to the compound having the chemical name 2-[(2S)-4-[7-(8-chloro-1-naphthyl)-2-[[(2S)-1-methylpyrrolidin-2-yl]methoxy]-6,8-dihydro-5H-pyrido[3,4-d]pyrimidin-4-yl]-1-(2-fluoroprop-2-enoyl)piperazin-2-yl]acetonitrile. The structural formula and preparation of adagrasib (MRTX849) are set forth in U.S. Pat. No. 1,068,9377, from Column 1286, line 32 to Column 1288, line 17, which are incorporated herein by reference.

[0036] MRTX1133 has the following structure: It refers to a compound having the chemical name 4-(4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-8-fluoro-2-(((2R,7aS)-2-fluorohexahydro-1H-pyrrolidin-7a-yl)methoxy)pyrodo[4,3-d]pyrimidin-7-yl)-5-ethynyl-6-fluoronaphthalen-2-ol. The structural formula and preparation method of MRTX1133 are described in WO2021041671.

[0037] The pharmacological activity of BI 1701963 was reported at the AACR Annual Meeting 2021#CT210; 2021 Apr 10-15 and May 17-21 (Abstract https: / / doi.org / 10.1158 / 1538-7445.AM2021-CT210).

[0038] The structural formula and preparation of BI-3406 are described in US Pat. No. 1,0898,487, which is incorporated herein by reference.

[0039] RG6330 is the same compound as GDC-6036. The structural formula and preparation of RG6330 are described in US Patent No. 1,236,068, which is incorporated herein by reference.

[0040] The structural formula and preparation method of JDQ443 are described in WO2021124222.

[0041] The structural formula and preparation of ARS-1620 is described in US Pat. No. 1,370,386, which is incorporated herein by reference.

[0042] The pharmacological activity of iExosomes has been reported in Nature volume 546, pages 498-503 (2017).

[0043] The structural formula and preparation of D-1553 are described in US Pat. No. 1,091,481, which is incorporated herein by reference.

[0044] The structural formula and preparation of BI 2852 are described in WO 2020 / 173935.

[0045] The structural formula and preparation method of SCH-54292 are reported in Bioorganic & Medicinal Chemistry, Volume 5, Issue 1, January 1997, Pages 125-133.

[0046] The structural formula and preparation of TLN-4601 are described in US Pat. No. 7,101,872, which is incorporated herein by reference.

[0047] The structural formula and method of preparation of Salirasib are described in US Reissue Patent No. RE39,682, which is incorporated herein by reference.

[0048] The structural formula and preparation of Deltarasin are described in US Pat. No. 9,861,623, which is incorporated herein by reference.

[0049] In the present disclosure, RAS inhibitors include sotorasib (AMG 510), adagrasib (MRTX849), MRTX1133), BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, D-1553, mRNA5671, BI 2852, SCH-54292, TLN-4601, Salirasib, Deltarasin, JAB-21822, BI 1823911, MK-1084, ELI-002, SDGR5, JAB-22000, ASP3082, and the like. In one embodiment, the compounds selected from the group consisting of sotorasib (AMG 510), adagrasib (MRTX849), MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, mRNA-5671, JAB-21822, BI 1823911, MK-1084, ELI-002, SDGR5, JAB-22000, and ASP3082, and in particular embodiments, sotorasib (AMG 510), adagrasib (MRTX849), and BI 1701963.

[0050] In one embodiment of the present disclosure, the RAS inhibitor is a KRAS inhibitor, an NRAS inhibitor, or an HRAS inhibitor. In another embodiment of the present disclosure, the RAS inhibitor is a KRAS inhibitor.

[0051] In the present disclosure, a KRAS inhibitor refers to an inhibitor that (i) blocks downstream signals caused by RAS by binding to KRAS, or (ii) inhibits protein-protein interaction between KRAS and a guanine nucleotide exchange factor (e.g., SOS1, SOS2). In addition, in the above (i), an inhibitor that binds to a GTP-binding mutant KRAS protein is a RAS-on inhibitor, and an inhibitor that binds to a GDP-binding mutant KRAS protein is a RAS-off inhibitor.

[0052] In one embodiment of the present disclosure, the KRAS inhibitor is sotorasib, adagrasib, MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, mRNA-5671, JAB-21822, BI 1823911, MK-1084, ELI-002, SDGR5, JAB-22000, and ASP3082.

[0053] In one embodiment of the present disclosure, RAS-on inhibitors are MRTX1133, RMC-6291, and RMC-6236. In one embodiment of the present disclosure, RAS-off inhibitors are sotorasib, adagrasib, MRTX1133, RG6330, LY3537982, JDQ443, ARS-1620, JNJ-74699157, JAB-21822, BI 1823911, MK-1084, and JAB-22000.

[0054] Examples of useful KRAS inhibitors include KRAS G12C inhibitors, KRAS G12D inhibitors, and pan-KRAS inhibitors, and in one embodiment, for example, a KRAS G12C inhibitor. Here, the KRAS G12C inhibitor refers to an inhibitor that binds to a KRAS mutant (e.g., protein, nucleic acid) having a mutation in which Gly at position 12 of KRAS is replaced by Cys and blocks downstream signaling caused by the mutant KRAS, and the KRAS G12D inhibitor refers to an inhibitor that binds to a KRAS mutant (e.g., protein, nucleic acid) having a mutation in which Gly at position 12 of KRAS is replaced by Asp and blocks downstream signaling caused by the mutant KRAS. A pan-KRAS inhibitor means (a) an inhibitor that binds to KRAS (e.g., protein, nucleic acid) regardless of the presence or absence of mutation, the position of mutation, or the amino acid after mutation, and blocks downstream signals caused by RAS, or (b) an inhibitor that binds to a guanine nucleotide exchange factor of KRAS (e.g., SOS1, SOS2) and inhibits the protein-protein interaction between KRAS and the guanine nucleotide exchange factor, thereby blocking downstream signals caused by RAS.

[0055] In one embodiment, the KRAS G12C inhibitors are sotorasib (AMG 510), adagrasib (MRTX849), RG6330, LY3537982, JDQ443, RMC-6291, JAB-21822, BI 1823911, and MK-1084, or in another embodiment, sotorasib (AMG 510) and adagrasib (MRTX849). In one embodiment, the KRAS G12D inhibitors are MRTX1133 and JAB-22000. In one embodiment, the pan-KRAS inhibitors are BI-3406, BI 1701963, LUNA18, SDGR5, and RMC-6236.

[0056] The RAS inhibitor according to the present disclosure can be administered by injection (intravenous injection, intra-arterial injection, local injection), oral administration, nasal administration, transdermal administration, pulmonary administration, or ophthalmic administration. Examples of injections include intravenous injection, subcutaneous injection, intradermal injection, intra-arterial injection, and local injection into target cells or organs. Examples of dosage forms of the RAS inhibitor for oral administration include tablets, powders, granules, syrups, capsules, and oral liquids. Examples of dosage forms of the RAS inhibitor for parenteral administration include injections, drip infusions, eye drops, ointments, suppositories, suspensions, poultices, lotions, aerosols, and plasters. In one embodiment, the RAS inhibitor is an injection or drip infusion. The RAS inhibitor according to the present disclosure can be formulated, for example, according to the methods described in the 18th Edition of the Japanese Pharmacopoeia (JP), the United States Pharmacopoeia (USP), or the European Pharmacopoeia (EP).

[0057] The dosage of the RAS inhibitor is not particularly limited, but is typically 0.1 to 5000 mg, 0.5 to 3000 mg, or 1.0 to 1000 mg per day when administered orally to an adult (body weight 60 kg) or child. This dosage can typically be administered once daily or over multiple days, or in 2 to 6 divided doses per day. The dosage and administration schedule can be modified when one or more additional chemotherapeutic agents are used. The administration schedule can be determined by the physician treating the particular patient. When sotorasib is administered as a single RAS inhibitor, it is orally administered at a dose selected from the group consisting of, for example, 960 mg, 480 mg, 240 mg, and 120 mg per day. The specific dosage, administration route, administration frequency, administration cycle, etc. of the RAS inhibitor are determined appropriately taking into account the type of disease to be treated, the patient's age, sex, weight, and severity of symptoms, other drugs to be administered in combination, etc.

[0058] In the present disclosure, tumors to be treated include, for example, solid cancers. Examples of solid cancers to be treated include non-small cell lung cancer, colon cancer, pancreatic cancer, mesothelioma, uterine cancer (e.g., cervical cancer and endometrial cancer), ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, testicular cancer, small intestine cancer, appendix cancer, and neuroendocrine tumors. In one embodiment, the solid cancer is at least one selected from non-small cell lung cancer, colon cancer, pancreatic cancer, ovarian cancer, and uterine cancer. In another embodiment, the solid cancer is at least one selected from non-small cell lung cancer, colon cancer, pancreatic cancer, uterine cancer, bile duct cancer, and bladder cancer. In a specific embodiment, the solid cancer is at least one selected from non-small cell lung cancer and colon cancer. In one embodiment, the tumor to be treated is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, and uterine cancer (e.g., cervical cancer, endometrial cancer). In another embodiment, the tumor to be treated is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, ovarian cancer, and bladder cancer, and in yet another embodiment, at least one selected from colorectal cancer, pancreatic cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, and uterine cancer (e.g., cervical cancer, endometrial cancer).

[0059] One embodiment of the present disclosure includes a pharmaceutical composition for tumor therapy (or tumor therapeutic agent) containing a compound represented by formula (I) and administered in combination with sotorasib, and the tumor to be treated is non-small cell lung cancer.

[0060] Another embodiment of the present disclosure includes a pharmaceutical composition for tumor therapy (or tumor therapeutic agent) containing a compound represented by formula (I) and administered in combination with sotorasib, wherein the tumor to be treated is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, and bladder cancer, and in a more specific embodiment, at least one selected from non-small cell lung cancer and colorectal cancer.

[0061] One embodiment of the present disclosure includes a pharmaceutical composition for tumor therapy (or a tumor therapeutic agent) containing a compound represented by formula (I) and administered in combination with adagrasib, wherein the tumor to be treated is non-small cell lung cancer.

[0062] Another embodiment of the present disclosure includes a pharmaceutical composition for tumor therapy (or a tumor therapeutic agent) containing a compound represented by formula (I) and administered in combination with adagrasib, wherein the tumor to be treated is at least one selected from non-small cell lung cancer and colorectal cancer.

[0063] One embodiment of the present disclosure includes a pharmaceutical composition for tumor therapy (or a tumor therapeutic agent) containing a compound represented by formula (I) and administered in combination with MRTX 1133, and in one embodiment, the tumor to be treated is at least one selected from non-small cell lung cancer, colorectal cancer, and pancreatic cancer. In another embodiment, the tumor to be treated by the pharmaceutical composition for tumor therapy (or a tumor therapeutic agent) containing a compound represented by formula (I) and administered in combination with MRTX 1133 is at least one selected from uterine cancer and bile duct cancer.

[0064] In one embodiment of the present disclosure, there is provided a method for treating tumors by administering a compound represented by formula (I) or a pharmaceutically acceptable salt thereof in combination with a RAS inhibitor. In one embodiment, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is administered orally.

[0065] One embodiment of the present disclosure includes a method for treating a tumor comprising orally administering to a patient a therapeutically effective amount of a compound of formula (I) in combination with sotorasib, wherein the tumor being treated is non-small cell lung cancer.

[0066] One embodiment of the present disclosure includes a method for treating a tumor comprising orally administering to a patient a therapeutically effective amount of a compound represented by formula (I) and sotorasib in combination, wherein the tumor to be treated is at least one selected from non-small cell lung cancer, colorectal cancer, pancreatic cancer, and bladder cancer, and in a more specific embodiment, at least one selected from non-small cell lung cancer and colorectal cancer.

[0067] One embodiment of the present disclosure includes a method for treating a tumor comprising orally administering to a patient a compound of formula (I) and adagrasib in therapeutically effective amounts, wherein the tumor to be treated is non-small cell lung cancer.

[0068] Another embodiment of the present disclosure includes a method for treating a tumor comprising orally administering to a patient a therapeutically effective amount of a compound of formula (I) and adagrasib, wherein the tumor to be treated is at least one selected from non-small cell lung cancer and colorectal cancer.

[0069] One embodiment of the present disclosure includes a method for treating tumors by orally administering to a patient a therapeutically effective amount of a compound represented by formula (I) in combination with MRTX 1133, wherein the tumor to be treated is at least one selected from non-small cell lung cancer, colorectal cancer, and pancreatic cancer. Another embodiment includes a method for treating tumors by orally administering to a patient a therapeutically effective amount of a compound represented by formula (I) in combination with MRTX 1133, wherein the tumor to be treated is at least one selected from uterine cancer and bile duct cancer.

[0070] The terms "comprising" or "including" as used herein are intended to be open-ended and inclusive, unless the context clearly dictates otherwise, and do not exclude additional, unrecited features, but rather encompass the closed terms "consisting of" or "consisting essentially of."

[0071] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by a person skilled in the art to which this invention belongs. Terms used herein should be interpreted as having a meaning consistent with the meaning in this specification and the related technical field, and should not be interpreted in an idealized or overly formal sense, unless otherwise defined.

[0072] Example 1 Antitumor effect of combined administration of the compound represented by formula (I) and a KRAS inhibitor in KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358

[0073] The combined growth inhibitory activity of the compound represented by formula (I) and sotorasib (manufactured by Angene International) was evaluated using KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358 (ATCC Cat. No. CRL-5807). The cells were cultured in RPMI 1640 medium (manufactured by FUJIFILM Wako Pure Chemicals) containing 10% v / v fetal bovine serum (FBS). 2 x 10 cells were plated in a 96-well plate. ３ The cells were seeded at 100 cells / well and incubated in 5% CO ２ The cells were cultured at 37°C for 24 hours. Subsequently, the cells were cultured for 72 hours using the compound represented by formula (I) (TOP 370 nM, 3-fold common ratio) and sotorasib (TOP 1 μM, 3-fold common ratio) under conditions of sotorasib alone and sotorasib and the compound represented by formula (I). The compound represented by formula (I) and sotorasib were adjusted to the designated concentrations by diluting DMSO stock with medium. For sotorasib alone, medium was added instead of the compound represented by formula (I). After 72 hours of culture, the cells were fixed with 10% trichloroacetic acid. The fixed cells were then stained with 0.4% (wt / vol) sulforhodamine B (SRB) dissolved in 1% acetic acid, and the absorbance at 515 nm was measured using a plate reader (PerkinElmer). The measured value was defined as the relative cell number. Using the cell count immediately before drug treatment and the cell count after 72 hours of drug treatment, the GI was calculated based on the concentration of each compound represented by formula (I) as 100%.５０ was calculated.

[0074] The compound represented by formula (I) was dissolved in 0.1 M hydrochloric acid, and sotorasib was dissolved in 1% Tween 80 (Sigma) / 2% HPMC (FUJIFILM Wako Pure Chemicals) / 97% distilled water (Otsuka Pharmaceutical Factory, Inc.) and used in the experiment.

[0075] Growth inhibition curve and GI ５０ The results are shown in Table 1 and Figure 1. In Table 1 and Figure 1, E7386 refers to the compound represented by formula (I). In Figure 1, the horizontal axis represents the concentration of sotorasib, and the vertical axis represents ΔT / C. In Figure 1, the measurement results are shown as the mean ± standard error (SEM) of ΔT / C. ΔT / C means the cell proliferation rate when sotorasib is administered alone (sotorasib group) relative to the cell proliferation rate when sotorasib and the compound represented by formula (I) are administered in combination. As a result, combined administration of the compound represented by formula (I) and sotorasib showed a significant growth inhibitory effect compared to sotorasib administered alone (sotorasib group) in a KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358 culture system.

[0076] Example 2 Antitumor effect of combined administration of the compound represented by formula (I) and a KRAS inhibitor in a transplant model of KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358

[0077] KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358 cells were added to Matrigel / HBSS (50:50) and plated at 8 × 10 ６ A cell suspension was prepared to obtain 1000 cells. The resulting suspension was transplanted subcutaneously into nude mice (CANN.Cg-Foxn1 nu / CrlCrlj, Charles River Japan). Six mice were included in each group. Starting on the seventh day after transplantation, the compound represented by formula (I) (25 mg / kg, once daily, 14 days, oral administration) and sotorasib (100 mg / kg, once daily, 14 days, oral administration) were administered alone or in combination to the single-administration group and the combined-administration group, respectively. The control group received neither of the above drugs.

[0078] At the time of administration, the compound of formula (I) was dissolved in 0.1 M hydrochloric acid, and sotorasib was dissolved in 1% Tween 80 (Sigma) / 2% HPMC (FUJIFILM Wako Pure Chemicals) / 97% distilled water (Otsuka Pharmaceutical Factory, Inc.) The administration started on the 1st day, and the major and minor diameters of the tumors that developed in each mouse were measured on the 4th, 8th, 11th, and 15th days using a Digimatic caliper (Mitutoyo Corp.).

[0079] Tumor volume was calculated according to the following formula: Tumor volume (mm ３ ) = tumor long diameter (mm) × tumor short diameter ２ (mm ２ ) / 2

[0080] The tumor volume measurements for each group are shown as the mean and standard error (SEM) in Table 2 and Figure 2. The numbers in Table 2 represent the mean value of tumor volume ± standard error (SEM).

[0081] As a result, combined administration of the compound represented by formula (I) and sotorasib showed a significant antitumor effect in a KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358 transplant model. In Table 2 and Figure 2, E7386 refers to the compound represented by formula (I). *** in Figure 2 indicates that combined administration of the compound represented by formula (I) and sotorasib statistically significantly inhibited tumor growth compared to administration of either compound alone (***: p<0.0001; Repeated measures ANOVA followed by Dunnett's type multiple comparison after logarithmic transformation).

[0082] Example 3 Cell proliferation inhibitory activity of the compound of formula (I) in combination with a KRAS inhibitor (sotorasib, adagrasib, MRTX1133, or BI3406) in KRAS G12C mutation-positive human pancreatic cancer cell line MIA PaCa-2, KRAS G12C mutation-positive human bladder cancer cell line UM-UC-3, KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358, KRAS G12D mutation-positive human cholangiocarcinoma cell line SNU-869, and KRAS G12D mutation-positive human uterine cancer cell line HEC-1-B

[0083] The growth inhibitory activity of the compound of formula (I) in combination with a KRAS inhibitor was evaluated using each KRAS mutation-positive cell line. The KRAS inhibitors used were sotorasib (manufactured by Angene International), adagrasib (manufactured by Amadis Chemical), or MRTX1133 (manufactured by WuXi AppTec). The source of each cell line and the culture medium are shown in Table 3. Cells were seeded at 5,000 cells / well in a 96-well flat-bottom ultra-low binding surface plate (manufactured by Corning) and cultured at 37°C with 5% CO2 for 1 day. Thereafter, the compound represented by formula (I) (TOP 333 nM, 3-fold common ratio) and one of three KRAS inhibitors, sotorasib (TOP 0.2-1 μM, 5-fold common ratio), adagrasib (TOP 1 μM, 5-fold common ratio), MRTX1133 (TOP 1 μM, 5-fold common ratio), or BI3406 (TOP 20 μM, 5-fold common ratio), were used and cultured for 5 days under the conditions of each single agent and in combination with the compound represented by formula (I). Note that each drug was prepared to the designated concentration by diluting DMSO stock with medium, and added so that the medium volume after drug addition was 100 μL / well. After 5 days of culture, 30 μL of CellTiter-Glo 3D Reagent (Promega) was added to each well and incubated for 30 minutes in the dark. 50 μL of each was then transferred to a separate white 96-well plate (Greiner), and the luciferase luminescence intensity was measured using a plate reader (PerkinElmer). The measured value was defined as the relative cell number. The cell number immediately before drug treatment and the cell number after 5 days of drug treatment were used to calculate the GI ratio, with the concentration of each compound represented by formula (I) as the 100% standard. ５０ was calculated.

[0084]

[0085] G.I. ５０The results and growth inhibition curves are shown in Table 4 and Figures 3A to 3F. In Table 4 and Figures 3A to 3F, E7386 refers to the compound represented by formula (I). In Figures 3A to 3F, the horizontal axis represents the concentration of each KRAS inhibitor, and the vertical axis represents ΔT / C. In Figures 3A to 3F, the measurement results for N = 3 (3 wells) for each drug addition condition are shown as the mean value of ΔT / C ± standard deviation (SD). ΔT / C refers to the cell growth rate when each KRAS inhibitor is administered alone relative to the cell growth rate when each KRAS inhibitor is administered in combination with E7386. As a result, in culture systems of KRAS G12C mutation-positive human pancreatic cancer cell line MIA PaCa-2 and KRAS G12C mutation-positive human bladder cancer cell line UM-UC-3, combination administration of E7386 and sotorasib showed significant growth inhibitory activity compared to sotorasib alone (Figures 3A and 3B).Furthermore, in culture systems of KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358, combination administration of E7386 and adagrasib showed significant growth inhibitory activity compared to adagrasib alone (Figure 3C). Furthermore, in culture systems of the KRAS G12D mutation-positive human cholangiocarcinoma cell line SNU-869 and the KRAS G12D mutation-positive human uterine cancer cell line HEC-1-B, combined administration of E7386 and MRTX1133 demonstrated significant growth inhibitory activity compared to MRTX1133 alone (Figures 3D and 3E).Furthermore, in culture systems of the KRAS G12C mutation-positive human pancreatic cancer cell line MIA PaCa-2, combined administration of E7386 and BI3406 demonstrated significant growth inhibitory activity compared to BI3406 alone (Figure 3F).

[0086]

[0087] Example 4 Antitumor effect of combined administration of E7386 and a KRAS inhibitor (sotrasib or adagrasib) in transplant models of KRAS G12C mutation-positive human colon cancer cell line SW837, KRAS G12C mutation-positive human pancreatic cancer cell line MIA PaCa-2, KRAS G12C mutation-positive human bladder cancer cell line UM-UC-3, and KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358

[0088] The efficacy test conditions for each cell line in mice (suspension medium, number of transplanted cells, mouse strain, administered drug, administration start date, and test period) are shown in Table 5. Each cell line was adjusted to the desired number of transplanted cells in the suspension medium, and the resulting suspension was transplanted subcutaneously into the lateral skin of nude mice (6-week-old, female). Starting from the administration start date shown in Table 5, the compound represented by formula (I) (25 mg / kg, once daily, oral administration), sotorasib (100 mg / kg, once daily, oral administration), and adagrasib (10 mg / kg, once daily, oral administration) were administered alone or in combination to the single-administration group or the combined-administration group until the day before the final day of the test period.

[0089] At the time of administration, E7386 was prepared in 0.1 M hydrochloric acid (FUJIFILM Wako Pure Chemicals), sotorasib was prepared in 1% Tween 80 (Tokyo Chemical Industry Co., Ltd.) / 2% HPMC (FUJIFILM Wako Pure Chemicals, Pure Chemicals) / 97% distilled water (Otsuka Pharmaceutical Factory, Inc.), and adagrasib was previously dissolved in 35% DMSO (FUJIFILM Wako Pure Chemicals) / 65% Tween 80 (Sigma) to a concentration of 100 mg / mL, and then prepared in 10% Captisol (Cyclolab) / 50 mM citrate buffer (MUTO PURE CHEMICALS). Starting from the first day of administration, the major and minor diameters of the tumors that developed in each mouse were measured twice a week until the final day of the test using a Digimatic caliper (Mitutoyo Corp.).

[0090] Tumor volume was calculated according to the following formula: Tumor volume (mm ３ ) = tumor long diameter (mm) × tumor short diameter ２ (mm ２ ) / 2

[0091] The tumor volume measurements for each group are shown in Table 6 and Figures 4A to 4D as the mean and standard error (SEM) (Figure 4A: SW837, Figure 4B: MIA PaCa-2, Figure 4C: UM-UC-3, Figure 4D: NCI-H358). The numbers in Table 6 represent the mean tumor volume ± standard error (SEM). Note that tests were performed using six mice per group. As a result, in KRAS G12C mutation-positive human colon cancer cell line SW837, KRAS G12C mutation-positive human pancreatic cancer cell line MIA PaCa-2, and KRAS G12C mutation-positive human bladder cancer cell line UM-UC-3 xenograft models, combined administration of E7386 and sotorasib demonstrated significant antitumor effects compared to administration of either drug alone. Furthermore, in a KRAS G12C mutation-positive human non-small cell lung cancer cell line NCI-H358 xenograft model, combined administration of E7386 and adagrasib demonstrated a significant antitumor effect compared to either compound administered alone. In Tables 5 and 6 and Figures 4A to 4D, E7386 refers to the compound represented by formula (I). In Figures 4A to 4D, * or **** indicates that combined administration of E7386 and sotorasib statistically significantly inhibited tumor growth compared to administration of either compound alone (*<0.05, ****: p<0.0001; Repeated measures ANOVA followed by Dunnett's type multiple comparison after logarithmic transformation).

[0092]

[0093]

Claims

1. A pharmaceutical composition for tumor treatment comprising (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is administered in combination with a RAS inhibitor. 【Chemistry 1】

2. A pharmaceutical composition for tumor treatment comprising a RAS inhibitor, and administered in combination with (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide represented by formula (I) or a pharmaceutically acceptable salt thereof. 【Chemistry 2】

3. 2. The pharmaceutical composition according to claim 1, wherein (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof is administered simultaneously or at different times with the RAS inhibitor.

4. 3. The pharmaceutical composition according to claim 2, wherein the RAS inhibitor and (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof are administered simultaneously or at different times.

5. (6S,9aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide or a pharmaceutically acceptable salt thereof aS)-N-benzyl-8-({6-[3-(4-ethylpiperazin-1-yl)azetidin-1-yl]pyridin-2-yl}methyl)-6-(2-fluoro-4-hydroxybenzyl)-4,7-dioxo-2-(prop-2-en-1-yl)hexahydro-2H-pyrazino[2,1-c][1,2,4]triazine-1(6H)-carboxamide.

6. The pharmaceutical composition according to claim 1 or 2, wherein the RAS inhibitor is a KRAS inhibitor.

7. The pharmaceutical composition according to claim 6, wherein the KRAS inhibitor is at least one selected from sotorasib, adagrasib, MRTX1133, BI-3406, BI 1701963, RG6330, LY3537982, JDQ443, RMC-6291, RMC-6236, LUNA18, ARS-1620, JNJ-74699157, GDC-6036, iExosomes, and mRNA-5671.

8. 7. The pharmaceutical composition of claim 6, wherein the KRAS inhibitor is sotorasib, adagrasib or BI 1701963.

9. The pharmaceutical composition of claim 6 , wherein the KRAS inhibitor is a KRAS G12C inhibitor.

10. The pharmaceutical composition of claim 9, wherein the KRAS G12C inhibitor is sotorasib or adagrasib.

11. 3. The pharmaceutical composition according to claim 1 or 2, wherein the tumor is at least one selected from non-small cell lung cancer, colon cancer, pancreatic cancer, mesothelioma, uterine cancer, ovarian cancer, bladder cancer, bile duct cancer, gastric cancer, breast cancer, small cell lung cancer, testicular cancer, small intestine cancer, appendix cancer, and neuroendocrine tumor.

12. 3. The pharmaceutical composition according to claim 1, wherein the tumor is at least one selected from non-small cell lung cancer, colon cancer, pancreatic cancer, uterine cancer, and ovarian cancer.

13. The pharmaceutical composition according to claim 1 or 2, wherein the tumor is at least one selected from non-small cell lung cancer and colorectal cancer.

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