CN118574632A - Medicine for treating or preventing cancer - Google Patents

Abstract

A drug for treating or preventing cancer, comprising a first active ingredient, used in combination with a second active ingredient, wherein the first active ingredient is a compound represented by the following formula (1) or a salt thereof or a solvate thereof, and the second active ingredient is a molecular targeting agent, [Chemical Formula 1]

Description

Drugs for the treatment or prevention of cancer

Technical Field

The present invention relates to a drug for treating or preventing cancer.

Background Art

Cancer is a disease in which cells proliferate uncontrollably due to abnormality in genes. Chemotherapy is used as a medical treatment for cancer, and various anticancer agents are applied. In recent years, many molecular targeted agents have been developed as anticancer agents, which specifically act on molecules specifically expressed in certain cancer cells or molecules that are hyperexpressed in cancer cells. As molecular targeted agents, for example, antibody drugs such as cetuximab, bevacizumab, and panitumumab are known (patent document 1), low molecular drugs such as gefitinib, erlotinib, and afatinib are known (patent document 2). Further, in recent years, medium molecule drugs (patent document 3) are also known.

Prior art literature

Patent Literature

Patent Document 1: Japanese Patent Application Publication No. 2018-076369

Patent Document 2: Japanese Patent Application Publication No. 2021-063014

Patent Document 3: International Publication No. 2021/090855

SUMMARY OF THE INVENTION

Problem that the invention aims to solve

Under such circumstances, regarding the treatment or prevention of cancer, there is a current demand for drugs with higher efficacy.

Therefore, an object of the present invention is to provide a drug that exhibits a more excellent effect than ever before in the treatment or prevention of cancer.

Means used to solve problems

The present invention includes the following [1] to [94].

[1] A drug for treating or preventing cancer, comprising a first active ingredient, used in combination with a second active ingredient, wherein the first active ingredient is a compound represented by the following formula (1) (hereinafter also referred to as "Compound 1") or a salt thereof or a solvate thereof, and the second active ingredient is a molecular targeting agent.

[Chemical formula 1]

[2] A drug for treating or preventing cancer, comprising a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is a molecular targeting agent and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

[3] A drug for treating or preventing cancer, comprising a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is a MAPK/ERK pathway inhibitor.

[4] A drug for treating or preventing cancer, comprising a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is a MAPK/ERK pathway inhibitor and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

[5] A drug for treating or preventing cancer, comprising a first active ingredient, used in combination with a second active ingredient, wherein the first active ingredient is compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is one selected from the group consisting of an EGFR inhibitor, a VEGF inhibitor, a SHP2 inhibitor, a KRAS-G12C inhibitor and a MEK inhibitor.

[6] A drug for treating or preventing cancer, comprising a first active ingredient, used in combination with a second active ingredient, wherein the first active ingredient is one selected from an EGFR inhibitor, a VEGF inhibitor, a SHP2 inhibitor, a KRAS-G12C inhibitor and a MEK inhibitor, and the second active ingredient is compound 1 or a salt thereof or a solvate thereof.

[7] The drug according to any one of [1] to [6], wherein the first active ingredient and the second active ingredient are provided as a kit.

[8] A method for treating or preventing cancer, comprising administering a first active ingredient and a second active ingredient to a subject, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is a molecular targeting agent.

[9] A method for treating or preventing cancer, comprising administering a first active ingredient and a second active ingredient to a subject, wherein the first active ingredient is a molecular targeting agent, and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

[10] A method for treating or preventing cancer, comprising administering a first active ingredient and a second active ingredient to a subject, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is a MAPK/ERK pathway inhibitor.

[11] A method for treating or preventing cancer, comprising administering a first active ingredient and a second active ingredient to a subject, wherein the first active ingredient is a MAPK/ERK pathway inhibitor, and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

[12] A method for treating or preventing cancer, comprising administering a first active ingredient and a second active ingredient to a subject, wherein the first active ingredient is compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is one selected from the group consisting of an EGFR inhibitor, a VEGF inhibitor, a SHP2 inhibitor, a KRAS-G12C inhibitor and a MEK inhibitor.

[13] A method for treating or preventing cancer, comprising administering a first active ingredient and a second active ingredient to a subject, wherein the first active ingredient is one selected from an EGFR inhibitor, a VEGF inhibitor, a SHP2 inhibitor, a KRAS-G12C inhibitor and a MEK inhibitor, and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

[14] A first active ingredient for use in the treatment or prevention of cancer, which is used in combination with a second active ingredient, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is a molecular targeting agent.

[15] A first active ingredient for use in the treatment or prevention of cancer, which is used in combination with a second active ingredient, wherein the first active ingredient is a molecular targeting agent and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

[16] A first active ingredient for use in the treatment or prevention of cancer, which is used in combination with a second active ingredient, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is a MAPK/ERK pathway inhibitor.

[17] A first active ingredient for use in the treatment or prevention of cancer, which is used in combination with a second active ingredient, wherein the first active ingredient is a MAPK/ERK pathway inhibitor and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

[18] A first active ingredient for use in the treatment or prevention of cancer, which is used in combination with a second active ingredient, wherein the first active ingredient is compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is one selected from the group consisting of an EGFR inhibitor, a VEGF inhibitor, a SHP2 inhibitor, a KRAS-G12C inhibitor and a MEK inhibitor.

[19] A first active ingredient for use in the treatment or prevention of cancer, which is used in combination with a second active ingredient, wherein the first active ingredient is one selected from EGFR inhibitors, VEGF inhibitors, SHP2 inhibitors, KRAS-G12C inhibitors and MEK inhibitors, and the second active ingredient is compound 1 or a salt thereof or a solvate thereof.

[20] Use of a first active ingredient in the preparation of a drug for treating or preventing cancer for use in combination with a second active ingredient, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is a molecular targeting agent.

[21] Use of a first active ingredient in the preparation of a drug for treating or preventing cancer for use in combination with a second active ingredient, wherein the first active ingredient is a molecular targeting agent and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

[22] Use of a first active ingredient in the preparation of a drug for treating or preventing cancer for use in combination with a second active ingredient, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is a MAPK/ERK pathway inhibitor.

[23] Use of a first active ingredient in the preparation of a drug for treating or preventing cancer for use in combination with a second active ingredient, wherein the first active ingredient is a MAPK/ERK pathway inhibitor and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

[24] Use of a first active ingredient in the preparation of a drug for the treatment or prevention of cancer used in combination with a second active ingredient, wherein the first active ingredient is compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is one selected from the group consisting of EGFR inhibitors, VEGF inhibitors, SHP2 inhibitors, KRAS-G12C inhibitors and MEK inhibitors.

[25] Use of a first active ingredient in the preparation of a drug for the treatment or prevention of cancer for use in combination with a second active ingredient, wherein the first active ingredient is selected from an EGFR inhibitor, a VEGF inhibitor, a SHP2 inhibitor, a KRAS-G12C inhibitor and a MEK inhibitor, and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

[26] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein the combination of the first active ingredient and the second active ingredient is administered simultaneously or separately (i.e., the first active ingredient and the second active ingredient are used in combination simultaneously or separately).

[27] The medicine, method, first active ingredient for use, or use according to any one of [1] to [26], wherein the first active ingredient is used in combination with the second active ingredient as a compound.

[28] The drug, method, first active ingredient for use, or use described in any one of [1], [2], [7], [8], [9], [14], [15], [20], [21], [26] and [27], wherein the molecular targeted agent is a MAPK/ERK pathway inhibitor.

[29] The drug, method, first active ingredient for use, or use described in any one of [1], [2], [7], [8], [9], [14], [15], [20], [21], [26], [27] and [28], wherein the molecular targeted agent is at least one selected from EGFR inhibitors, VEGF inhibitors, SHP2 inhibitors, KRAS-G12C inhibitors and MEK inhibitors.

[30] The drug, method, first active ingredient for use, or use described in any one of [1], [2], [7], [8], [9], [14], [15], [20], [21], [26], [27], [28] and [29], wherein the molecular targeted agent is an EGFR inhibitor, a VEGF inhibitor, a SHP2 inhibitor, a KRAS-G12C inhibitor or a MEK inhibitor.

[31] The drug, method, first active ingredient for use, or use described in any one of [1], [2], [7], [8], [9], [14], [15], [20], [21], [26], [27], [28], [29] and [30], wherein the molecular targeted agent is an EGFR inhibitor.

[32] The drug, method, first active ingredient for use, or use described in any one of [1], [2], [7], [8], [9], [14], [15], [20], [21], [26], [27], [28], [29] and [30], wherein the molecular targeting agent is a VEGF inhibitor.

[33] The drug, method, first active ingredient for use, or use described in any one of [1], [2], [7], [8], [9], [14], [15], [20], [21], [26], [27], [28], [29] and [30], wherein the molecular targeted agent is a SHP2 inhibitor.

[34] The drug, method, first active ingredient for use, or use described in any one of [1], [2], [7], [8], [9], [14], [15], [20], [21], [26], [27], [28], [29] and [30], wherein the molecular targeted agent is a KRAS-G12C inhibitor.

[35] The drug, method, first active ingredient for use, or use described in any one of [1], [2], [7], [8], [9], [14], [15], [20], [21], [26], [27], [28], [29] and [30], wherein the molecular targeted agent is a MEK inhibitor.

[36] The drug, method, first active ingredient for use, or use described in [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27], [29], [30] and [31], wherein the EGFR inhibitor is at least one selected from gefitinib, erlotinib, afatinib, osimertinib, lapatinib or their salts or solvates, and anti-EGFR antibodies.

[37] A drug, method, first active ingredient for use, or use according to any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27], [29], [30], [31], and [36], wherein the EGFR inhibitor is an anti-EGFR antibody.

[38] The drug, method, first active ingredient for use, or use described in [36] or [37], wherein the anti-EGFR antibody has a heavy chain variable region comprising the amino acid sequence of sequence number 1, and a light chain variable region comprising the amino acid sequence of sequence number 2.

[39] The drug, method, first active ingredient for use, or use according to any one of [36] to [38], wherein the anti-EGFR antibody is cetuximab.

[40] The drug, method, first active ingredient for use, or use described in any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27] and [29] to [39], wherein the VEGF inhibitor is at least one selected from sorafenib, pazopanib, sunitinib, axitinib, regorafenib or their salts or solvates, and anti-VEGF antibodies.

[41] The drug, method, first active ingredient for use, or use described in any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27] and [29] to [40], wherein the VEGF inhibitor is an anti-VEGF antibody.

[42] The drug, method, first active ingredient for use, or use described in [40] or [41], wherein the anti-VEGF antibody has a heavy chain variable region comprising the amino acid sequence of sequence number 3, and a light chain variable region comprising the amino acid sequence of sequence number 4.

[43] The drug, method, first active ingredient for use, or use according to any one of [40] to [42], wherein the anti-VEGF antibody is bevacizumab.

[44] A drug, method, first active ingredient for use, or use according to any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27], and [29] to [43], wherein the SHP2 inhibitor is at least one selected from RMC4550, TNO155, RLY1971, SHP099, NSC-87877, and their salts and solvates thereof.

[45] A drug, method, first active ingredient for use, or use according to any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27], and [29] to [44], wherein the SHP2 inhibitor is at least one selected from RMC4550, TNO155, and their salts and solvates thereof.

[46] A drug, method, first active ingredient for use, or use according to any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27], and [29] to [45], wherein the SHP2 inhibitor is RMC4550 or a salt thereof or a solvate thereof.

[46-1] A drug, method, first active ingredient for use, or use according to any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27], and [29] to [45], wherein the SHP2 inhibitor is TNO155 or a salt thereof or a solvate thereof.

[47] A drug, method, first active ingredient for use, or use according to any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27], and [29] to [46], wherein the KRAS-G12C inhibitor is at least one selected from sotolacib, MRTX849, and salts thereof and solvates thereof.

[48] The drug, method, first active ingredient for use, or use described in any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27] and [29] to [47], wherein the KRAS-G12C inhibitor is sotolacib.

[48-1] A drug, method, first active ingredient for use, or use described in any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27], and [29] to [47], wherein the KRAS-G12C inhibitor is MRTX849.

[49] The drug, method, first active ingredient for use, or use described in any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27] and [29] to [47], wherein the KRAS-G12C inhibitor is sotolacib or a salt thereof or a solvate thereof.

[49-1] A drug, method, first active ingredient for use, or use described in any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27] and [29] to [47], wherein the KRAS-G12C inhibitor is MRTX849 or a salt thereof or a solvate thereof.

[50] A drug, method, first active ingredient for use, or use according to any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27], and [29] to [49], wherein the MEK inhibitor is at least one selected from the group consisting of a compound represented by the following formula (2), trametinib, selumetinib, CH4987655, and salts thereof, and solvates thereof.

[Chemical formula 2]

[51] The drug, method, first active ingredient for use, or use described in any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27] and [29] to [50], wherein the MEK inhibitor is a compound represented by the following formula (2), trametinib, or a salt thereof, or a solvate thereof.

[Chemical formula 3]

[52] A drug, method, first active ingredient for use, or use according to any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27], and [29] to [51], wherein the MEK inhibitor is a compound represented by the following formula (2) or a salt thereof or a solvate thereof.

[53] The drug, method, first active ingredient for use, or use described in any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27] and [29] to [51], wherein the MEK inhibitor is trametinib or a salt thereof or a solvate thereof.

[54] The drug, method, first active ingredient for use, or use described in any one of [5], [6], [7], [12], [13], [18], [19], [24], [25], [26], [27], [29] to [51] and [53], wherein the MEK inhibitor is trametinib dimethyl sulfoxide adduct.

[55] The drug, method, first active ingredient for use, or use according to any one of [1] to [54], wherein Compound 1 or a salt thereof or a solvate thereof is a solvate of Compound 1.

[56] The medicine, method, first active ingredient for use, or use described in [55], wherein the solvate is a hydrate.

[57] The drug, method, first active ingredient for use, or use according to any one of [1] to [54], wherein Compound 1 or a salt thereof or a solvate thereof is Compound 1.

[58] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is a hydrate of Compound 1, and the molecular targeting agent, the MAPK/ERK pathway inhibitor or the EGFR inhibitor is cetuximab.

[59] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is a hydrate of Compound 1, and the molecular targeting agent, the MAPK/ERK pathway inhibitor or the VEGF inhibitor is an anti-VEGF antibody.

[60] The drug, method, first active ingredient for use, or use described in [59], wherein the anti-VEGF antibody is bevacizumab.

[61] A drug, method, first active ingredient for use, or use according to any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is a hydrate of Compound 1, and the molecular targeted agent, the MAPK/ERK pathway inhibitor or the SHP2 inhibitor is RMC4550 or its salt or solvate thereof.

[61-1] A drug, method, first active ingredient for use, or application described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is a hydrate of Compound 1, and the molecular targeting agent, the MAPK/ERK pathway inhibitor or the SHP2 inhibitor is TNO155 or its salt or solvate thereof.

[62] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is a hydrate of Compound 1, and the above-mentioned molecular targeted agent, the above-mentioned MAPK/ERK pathway inhibitor or the above-mentioned SHP2 inhibitor is RLY1971 or its salt or solvate thereof.

[63] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is a hydrate of Compound 1, and the molecular targeted agent, the MAPK/ERK pathway inhibitor or the KRAS-G12C inhibitor is sotolacib or its salt or solvate thereof.

[63-1] A drug, method, first active ingredient for use, or application described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is a hydrate of Compound 1, and the above-mentioned molecular targeted agent, the above-mentioned MAPK/ERK pathway inhibitor or the above-mentioned KRAS-G12C inhibitor is MRTX849 or its salt or solvate thereof.

[64] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is a hydrate of Compound 1, and the above-mentioned molecular targeting agent, the above-mentioned MAPK/ERK pathway inhibitor or the above-mentioned MEK inhibitor is a compound represented by the following formula (2) or its salt or solvate thereof.

[65] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is a hydrate of Compound 1, and the above-mentioned molecular targeted agent, the above-mentioned MAPK/ERK pathway inhibitor or the above-mentioned MEK inhibitor is trametinib or its salt or solvate thereof.

[66] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate is Compound 1, and the molecular targeted agent, the MAPK/ERK pathway inhibitor or the EGFR inhibitor is cetuximab.

[67] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate is Compound 1, and the molecular targeting agent, the MAPK/ERK pathway inhibitor or the VEGF inhibitor is an anti-VEGF antibody.

[68] The drug, method, first active ingredient for use, or use described in [67], wherein the anti-VEGF antibody is bevacizumab.

[69] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is Compound 1, and the above-mentioned molecular targeted agent, the above-mentioned MAPK/ERK pathway inhibitor or the above-mentioned SHP2 inhibitor is RMC4550 or its salt or solvate thereof.

[69-1] A drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or a salt thereof or a solvate thereof is Compound 1, and the molecular targeting agent, the MAPK/ERK pathway inhibitor or the SHP2 inhibitor is TNO155 or a salt thereof or a solvate thereof.

[70] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is Compound 1, and the above-mentioned molecular targeted agent, the above-mentioned MAPK/ERK pathway inhibitor or the above-mentioned SHP2 inhibitor is RLY1971 or its salt or solvate thereof.

[71] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is Compound 1, and the molecular targeted agent, the MAPK/ERK pathway inhibitor or the KRAS-G12C inhibitor is sotolacib or its salt or solvate thereof.

[71-1] A drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or a solvate thereof is Compound 1, and the molecular targeted agent, the MAPK/ERK pathway inhibitor or the KRAS-G12C inhibitor is MRTX849 or its salt or a solvate thereof.

[72] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is Compound 1, and the molecular targeting agent, the MAPK/ERK pathway inhibitor or the MEK inhibitor is a compound represented by formula (2) or its salt or solvate thereof.

[73] The drug, method, first active ingredient for use, or use described in any one of [1] to [25], wherein Compound 1 or its salt or solvate thereof is Compound 1, and the above-mentioned molecular targeted agent, the above-mentioned MAPK/ERK pathway inhibitor or the above-mentioned MEK inhibitor is trametinib or its salt or solvate thereof.

[74] The drug, method, first active ingredient for use, or use according to any one of [1] to [73], wherein the cancer is a solid cancer or a blood cancer.

[75] A drug, method, first active ingredient for use, or application according to any one of [1] to [74], wherein the cancer is at least one selected from lung cancer, esophageal cancer, gastric cancer, colorectal cancer, uterine cancer, ovarian cancer, pancreatic cancer, bladder cancer, thyroid cancer, skin cancer, leukemia, malignant lymphoma, and multiple myeloma.

[76] A drug, method, first active ingredient for use, or application described in any one of [1] to [75], wherein the cancer is a cancer associated with abnormalities in the RAS gene or MAPK/ERK pathway (abnormalities in proteins on the MAPK/ERK pathway or genes that produce the proteins).

[77] The drug, method, first active ingredient for use, or use according to any one of [1] to [76], wherein the abnormality in the MAPK/ERK pathway is abnormal activation of the MAPK/ERK pathway.

[78] The drug, method, first active ingredient for use, or use described in any one of [1] to [77], wherein the abnormality in the MAPK/ERK pathway is abnormal activation caused by amplification of a protein in the MAPK/ERK pathway.

[79] The drug, method, first active ingredient for use, or use described in any one of [1] to [78], wherein the abnormality in the MAPK/ERK pathway is abnormal activation caused by a gene mutation in the MAPK/ERK pathway.

[80] The drug, method, first active ingredient for use, or use according to any one of [1] to [79], wherein the cancer is a cancer associated with abnormality in the RAS gene.

[81] The drug, method, first active ingredient for use, or use described in [80], wherein the abnormality of the RAS gene is a mutation in the coding region of the RAS gene and/or an amplification of the copy number of the RAS gene.

[82] The drug, method, first active ingredient for use, or use described in any one of [76] to [81], wherein the RAS gene is at least one RAS gene selected from the group consisting of KRAS gene, NRAS gene, and HRAS gene.

[83] The drug, method, first active ingredient for use, or use described in any one of [76] to [82], wherein the RAS gene is the KRAS gene.

[84] The drug, method, first active ingredient for use, or use according to any one of [1] to [83], wherein the cancer is associated with the production of mutant RAS protein and/or increased production of RAS protein.

[85] The drug, method, first active ingredient for use, or use according to any one of [1] to [84], wherein the cancer is associated with the production of a mutant RAS protein.

[86] The drug, method, first active ingredient for use, or use described in [84] or [85], wherein the mutant RAS protein is one or more mutant RAS proteins selected from mutant KRAS protein, mutant NRAS protein, and mutant HRAS protein.

[87] The drug, method, first active ingredient for use, or use described in any one of [84] to [86], wherein the mutant RAS protein has a mutation at at least one amino acid position selected from G12, G13 and Q61.

[88] A drug, method, first active ingredient for use, or use according to any one of [84] to [87], wherein the mutant RAS protein has a mutation at at least one amino acid position selected from G12, G13 and Q61 (but excluding mutations in G12D).

[89] The drug, method, first active ingredient for use, or use according to any one of [84] to [88], wherein the mutant RAS protein is a mutant KRAS protein.

[90] The drug, method, first active ingredient for use, or use described in any one of [84] to [89], wherein the mutant RAS protein is a mutant KRAS protein having at least one amino acid mutation selected from G12A, G12C, G12D, G12S, G12V, G13D, Q61H, and Q61K.

[91] The drug, method, first active ingredient for use, or use described in any one of [84] to [90], wherein the mutant RAS protein is a mutant KRAS protein having at least one amino acid mutation selected from G12A, G12C, G12S, G12V, G13D, Q61H, and Q61K.

[92] The drug, method, first active ingredient for use, or use described in any one of [84] to [88], wherein the mutant RAS protein is a mutant NRAS protein having at least one amino acid mutation selected from G12C, G12D, G13D, G13V, Q61K, and Q61L.

[93] The drug, method, first active ingredient for use, or use described in any one of [84] to [88] and [92], wherein the mutant RAS protein is a mutant NRAS protein having at least one amino acid mutation selected from G12C, G13D, G13V, Q61K, and Q61L.

[94] The drug, method, first active ingredient for use, or use described in any one of [84] to [88], wherein the mutant RAS protein is a mutant HRAS protein having an amino acid mutation of G13R.

Effects of the Invention

According to the present invention, a drug is provided that exhibits a more excellent effect than ever before in the treatment or prevention of cancer.

BRIEF DESCRIPTION OF THE DRAWINGS

[Figure 1] Figure 1 is a graph showing the results of evaluating the RAS signaling inhibitory activity of the combination of Compound 1 and Sotolacib using NCI-H358 in Example 1. More specifically, Figure 1 is an electrophoretic diagram showing the results of Western blotting of proteins (KRAS, NRAS, HRAS, KRAS-GTP, NRAS-GTP, HRAS-GTP, ERK, AKT, MEK, pERK, pAKT, and pMEK) extracted from NCI-H358.

[Fig. 2] Fig. 2 is a graph showing the results of the evaluation of the RAS signaling inhibitory activity by the combination of Compound 1 and Sotolacib using NCI-H1373 in Example 1. More specifically, Fig. 2 is an electrophoretic diagram showing the results of Western blotting of proteins (KRAS, NRAS, HRAS, KRAS-GTP, NRAS-GTP, HRAS-GTP, ERK, AKT, MEK, pERK, pAKT, and pMEK) extracted from NCI-H1373.

[Fig. 3] Fig. 3 is a diagram showing the results of evaluating the RAS signaling inhibitory activity by combining Compound 1 with RMC-4550 (RMC) using NCI-H441 in Example 1. More specifically, Fig. 3 is an electrophoretic diagram showing the results of Western blotting of proteins (KRAS, NRAS, HRAS, KRAS-GTP, NRAS-GTP, HRAS-GTP, ERK, AKT, MEK, pERK, pAKT, and pMEK) extracted from NCI-H441.

[Fig. 4] Fig. 4 is a diagram showing the results of evaluating the RAS signaling inhibitory activity by combining Compound 1 with trametinib (trame) using NCI-H441 in Example 1. More specifically, Fig. 4 is an electrophoretic diagram showing the results of Western blotting of proteins (KRAS, NRAS, HRAS, KRAS-GTP, NRAS-GTP, HRAS-GTP, ERK, AKT, MEK, pERK, pAKT, and pMEK) extracted from NCI-H441.

[Fig. 5] Fig. 5 is a graph showing the results of the evaluation of the RAS signaling inhibitory activity by the combination of Compound 1 and Compound 2 using NCI-H358 in Example 1. More specifically, Fig. 5 is an electrophoretic diagram showing the results of Western blotting of proteins (KRAS, KRAS-GTP, ERK, AKT, MEK, pERK, pAKT, and pMEK) extracted from NCI-H358.

[Fig. 6] Fig. 6 is a graph showing the results of the evaluation of the RAS signaling inhibitory activity by the combination of Compound 1 and Compound 2 using NCI-H441 in Example 1. More specifically, Fig. 6 is an electrophoretic diagram showing the results of Western blotting of proteins (KRAS, KRAS-GTP, ERK, AKT, MEK, pERK, pAKT, and pMEK) extracted from NCI-H441.

[Figure 7] Figure 7 is a graph showing the results of evaluating the cell proliferation inhibitory activity of the combination of compound 1 and RMC-4550 using NCI-H358 in Example 2. Respectively, the horizontal axis of Figure 7a represents the concentration of RMC-4550, and the vertical axis represents the concentration of compound 1. Each cell in Figure 7a shows GI (growth inhibition, proliferation inhibition rate (%)) at its respective concentration. The horizontal axis of Figure 7b represents the value of the concentration of RMC-4550 divided by the concentration of GI155 (0.55 μM) given as a single dose of RMC-4550, and the vertical axis represents the value of the concentration of compound 1 divided by the concentration of GI155 (0.033 μM) given as a single dose of compound 1. The concentration of GI155 administered in the case of the combination of RMC-4550 and compound 1 was measured and plotted in the graph of Figure 7b.

[Figure 8] Figure 8 is a graph showing the results of evaluating the cell proliferation inhibitory activity of the combination of compound 1 and RMC-4550 using NCI-H441 in Example 2. The horizontal axis of Figure 8a represents the concentration of RMC-4550, and the vertical axis represents the concentration of compound 1. Each cell in Figure 8a shows GI (%) at its respective concentration. The horizontal axis of Figure 8b represents the value of the concentration of RMC-4550 divided by the concentration of GI100 (0.35 μM) given as a single dose of RMC-4550, and the vertical axis represents the value of the concentration of compound 1 divided by the concentration of GI100 (0.0055 μM) given as a single dose of compound 1. The concentration of GI100 given in the case of the combination of RMC-4550 and compound 1 was measured and plotted in the graph of Figure 8b.

[Figure 9] Figure 9 is a graph showing the results of evaluating the cell proliferation inhibitory activity of the combination of compound 1 and trametinib in Example 2 using NCI-H358. The horizontal axis of Figure 9a represents the concentration of trametinib, and the vertical axis represents the concentration of compound 1. Each cell in Figure 9a shows GI (%) at each concentration. The horizontal axis of Figure 9b represents the concentration of trametinib divided by the concentration of GI50 (0.00035 μM) given with a single dose of trametinib, and the vertical axis represents the concentration of compound 1 divided by the concentration of GI50 (0.0046 μM) given with a single dose of compound 1. The concentration of GI50 given in the case of combining trametinib and compound 1 was measured and plotted in the graph of Figure 9b.

[ Fig. 10] Fig. 10 is a graph showing the results of in vivo antitumor activity evaluation of the combination of Compound 1 and cetuximab using LoVo in Example 3. The vertical axis represents tumor volume, and the horizontal axis represents the number of days after cell transplantation.

[Fig. 11] Fig. 11 is a diagram showing the results of the in vivo RAS signaling inhibitory activity evaluation of Example 4. "Cetu" in the figure means cetuximab. More specifically, Fig. 11 is an electrophoretic diagram showing the results of Western blotting of proteins (KRAS, KRAS-GTP, ERK, AKT, EGFR, pERK, pAKT, and pEGFR) extracted from tumor cells of Example 4.

[ Fig. 12] Fig. 12 is a graph showing the results of in vivo antitumor activity evaluation of the combination of Compound 1 and bevacizumab using AsPC-1 (pancreatic cancer: KRAS G12D mutant) in Example 5. The vertical axis represents tumor volume, and the horizontal axis represents the number of days after cell transplantation.

[ Fig. 13] Fig. 13 is a graph showing the results of evaluating the cell proliferation inhibitory activity of the combination of Compound 1 and Sotolacib using SW837 in Example 6.

[ Fig. 14] Fig. 14 is a graph showing the results of evaluating the cell proliferation inhibitory activity of the combination of Compound 1 and MRTX849 in Example 7 using HCC-1171 and NCI-H1373.

[ Fig. 15] Fig. 15 is a graph showing the results of evaluating the cell proliferation inhibitory activity of the combination of Compound 1 and TNO155 in Example 7 using HCC-1171, NCI-H23, NCI-H1373 and UM-UC-3.

[ Fig. 16] Fig. 16 is a graph showing the results of in vivo antitumor activity evaluation of the combination of Compound 1 and sotolacib using CTG-2579 (lung cancer: KRAS G12C mutant) in Example 8. The vertical axis represents tumor volume, and the horizontal axis represents the number of days elapsed from the start of administration, with the administration start date being day 0.

[ Fig. 17] Fig. 17 is a diagram showing the results of the in vivo RAS signaling inhibitory activity evaluation in Example 9. More specifically, Fig. 17 is an electrophoretic diagram showing the results of Western blotting of proteins (KRAS, KRAS-GTP, ERK, pERK, AKT, and pAKT) extracted from tumor cells in Example 9.

[ Fig. 18 ] Fig. 18 is a graph showing the results of in vivo antitumor activity evaluation of the combination of Compound 1 and MRTX849 using LU65 (lung cancer: KRAS G12C mutant) and UM-UC-3 (bladder cancer KRAS G12C mutant) in Example 10. The vertical axis represents tumor volume, and the horizontal axis represents the number of days after cell transplantation.

[ Fig. 19] Fig. 19 is a graph showing the results of in vivo antitumor activity evaluation of the combination of Compound 1 and TNO155 using NCI-H1373 (lung cancer: KRAS G12C mutant) in Example 11. The vertical axis represents tumor volume, and the horizontal axis represents the number of days elapsed from the start of administration, with the administration start date being day 0.

[ Fig. 20] Fig. 20 is a graph showing the results of in vivo antitumor activity evaluation of the combination of Compound 1 and Compound 2 using NCI-H441 (lung cancer: KRAS G12V mutant) in Example 12. The vertical axis represents tumor volume, and the horizontal axis represents the number of days after cell transplantation.

[ Fig. 21] Fig. 21 is a graph showing the results of in vivo antitumor activity evaluation of the combination of Compound 1 and Compound 2 using NCI-H1373 (lung cancer: KRAS G12C mutant) in Example 12. The vertical axis represents tumor volume, and the horizontal axis represents the number of days elapsed from the start of administration, with the administration start date being day 0.

[ Fig. 22] Fig. 22 is a diagram showing the results of the in vivo RAS signaling inhibitory activity evaluation in Example 13. More specifically, Fig. 22 is an electrophoretic diagram showing the results of Western blotting of proteins (KRAS, KRAS-GTP, ERK, pERK, AKT, pAKT, MEK, and pMEK) extracted from tumor cells in Example 13.

Modes for carrying out the invention

The embodiments of the present invention are described. However, the present invention is not limited to the following embodiments. The therapeutic or preventive drugs and therapeutic or preventive methods of the present invention can be administered or applied to humans. In this specification, "～" indicating a range includes values at both ends, for example, "A～B" means a range above A and below B. In this specification, the term "about" means a range of +10% and -10% of the value of the numerical value when used in combination with a numerical value. In the present invention, the meaning of the term "and/or" includes all combinations of appropriate combinations of "and" and "or". Specifically, for example, "A, B, and/or C" includes the following 7 variations: (i) A, (ii) B, (iii) C, (iv) A and B, (v) A and C, (vi) B and C, (vii) A, B and C.

One embodiment of the present invention is a drug for treating or preventing cancer, which comprises a first active ingredient, used in combination with a second active ingredient, wherein the first active ingredient is a compound represented by the following formula (1) (hereinafter also referred to as "Compound 1") or its salt or solvate thereof, and the second active ingredient is a molecular targeting agent.

[Chemical formula 4]

Another embodiment of the present invention is a drug for treating or preventing cancer, which comprises a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is a molecular targeting agent and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

In the manner of using compound 1 or its salt or its solvate in combination with a molecular targeting agent, compound 1 or its salt or its solvate of the present invention can be used as any of the first active ingredient or the second active ingredient, and the molecular targeting drug of the present invention can be used as any of the first active ingredient or the second active ingredient. Preferably, the molecular targeting agent is a MAPK/ERK pathway inhibitor. More preferably, the molecular targeting agent is one selected from EGFR inhibitors, VEGF inhibitors, SHP2 inhibitors, KRAS-G12C inhibitors and MEK inhibitors.

One embodiment of the present invention is a drug for treating or preventing cancer, which comprises a first active ingredient, used in combination with a second active ingredient, wherein the first active ingredient is compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is a MAPK/ERK pathway inhibitor.

Another embodiment of the present invention is a drug for treating or preventing cancer, which comprises a first active ingredient, used in combination with a second active ingredient, wherein the first active ingredient is a MAPK/ERK pathway inhibitor, and the second active ingredient is compound 1 or a salt thereof or a solvate thereof.

In the manner of using compound 1 or its salt or their solvate in combination with a MAPK/ERK pathway inhibitor, compound 1 or its salt or their solvate described in this embodiment can be used as either the first active ingredient or the second active ingredient, and the MAPK/ERK pathway inhibitor described in this embodiment can be used as either the first active ingredient or the second active ingredient.

One embodiment of the present invention is a drug for treating or preventing cancer, which comprises a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is an EGFR inhibitor.

Another embodiment of the present invention is a drug for treating or preventing cancer, which comprises a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is an EGFR inhibitor and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

In the manner of using compound 1 or its salt or its solvate in combination with an EGFR inhibitor, compound 1 or its salt or its solvate described in this embodiment can be used as any of the first active ingredient or the second active ingredient, and the EGFR inhibitor described in this embodiment can be used as any of the first active ingredient or the second active ingredient. The EGFR inhibitor can be at least one selected from gefitinib, erlotinib, afatinib, osimertinib, lapatinib, or their salts or their solvates and anti-EGFR antibodies, and can be an anti-EGFR antibody. Preferably, the EGFR inhibitor can be an anti-EGFR antibody. The anti-EGFR antibody can have a heavy chain variable region comprising the amino acid sequence of sequence number 1 and a light chain variable region comprising the amino acid sequence of sequence number 2, and can be cetuximab. More preferably, the EGFR inhibitor can be cetuximab.

One embodiment of the present invention is a drug for treating or preventing cancer, which comprises a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is a VEGF inhibitor.

Another embodiment of the present invention is a drug for treating or preventing cancer, which comprises a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is a VEGF inhibitor and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

In the form of a combination of compound 1 or its salt or its solvate with a VEGF inhibitor, compound 1 or its salt or its solvate described in this embodiment can be used as either the first active ingredient or the second active ingredient, and the VEGF inhibitor described in this embodiment can be used as either the first active ingredient or the second active ingredient. The VEGF inhibitor can be at least one selected from sorafenib, pazopanib, sunitinib, axitinib, regorafenib, or their salts or their solvates, and anti-VEGF antibodies, and can be an anti-VEGF antibody, can have a heavy chain variable region comprising the amino acid sequence of sequence number 3 and a light chain variable region comprising the amino acid sequence of sequence number 4, and can be bevacizumab, ramucirumab or aflibercept. As a VEGF inhibitor, an anti-VEGF antibody is preferred, and bevacizumab is more preferred.

One embodiment of the present invention is a drug for treating or preventing cancer, comprising a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is a SHP2 inhibitor.

Another embodiment of the present invention is a drug for treating or preventing cancer, which comprises a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is a SHP2 inhibitor and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

In the combination of compound 1 or its salt or its solvate and SHP2 inhibitor, compound 1 or its salt or its solvate described in this embodiment can be used as either the first active ingredient or the second active ingredient, and the SHP2 inhibitor described in this embodiment can be used as either the first active ingredient or the second active ingredient. The SHP2 inhibitor can be at least one selected from RMC4550, TNO155, RLY1971, SHP099, NSC-87877 and their salts and their solvates. Preferably, as the SHP2 inhibitor, RMC4550, RLY1971, TNO155 or their salts or their solvates can be listed. More preferably, the SHP2 inhibitor is RMC4550, TNO155 or their salts or their solvates.

One embodiment of the present invention is a drug for treating or preventing cancer, comprising a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is an inhibitor of KRAS-G12C.

Another embodiment of the present invention is a drug for treating or preventing cancer, comprising a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is a KRAS-G12C inhibitor and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

In the manner of using compound 1 or its salt or its solvate in combination with a KRAS-G12C inhibitor, compound 1 or its salt or its solvate described in this embodiment can be used as either the first active ingredient or the second active ingredient, and the KRAS-G12C inhibitor described in this embodiment can be used as either the first active ingredient or the second active ingredient. The KRAS-G12C inhibitor can be at least one selected from sotolacib, MRTX849, and their salts and solvates thereof, preferably sotolacib, MRTX849, or their salts or solvates thereof.

One embodiment of the present invention is a drug for treating or preventing cancer, comprising a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is Compound 1 or a salt thereof or a solvate thereof, and the second active ingredient is a MEK inhibitor.

Another embodiment of the present invention is a drug for treating or preventing cancer, comprising a first active ingredient in combination with a second active ingredient, wherein the first active ingredient is a MEK inhibitor and the second active ingredient is Compound 1 or a salt thereof or a solvate thereof.

In the manner of using compound 1 or its salt or its solvate in combination with a MEK inhibitor, compound 1 or its salt or its solvate described in the present embodiment can be used as any of the first active ingredient or the second active ingredient, and the MEK inhibitor described in the present embodiment can be used as any of the first active ingredient or the second active ingredient. As a MEK inhibitor, at least one selected from the following compounds represented by formula (2), trametinib, selumetinib, CH4987655 and their salts and their solvates can be listed, preferably, compounds represented by formula (2), trametinib or their salts or their solvates can be listed. More preferably, as a MEK inhibitor, compounds represented by formula (2) or their salts or their solvates can be listed.

The first active ingredient and the second active ingredient may be provided as a kit. A further embodiment of the present invention may also be referred to as a kit comprising the first active ingredient and the second active ingredient. The drug of the present invention may be provided as a kit. Another embodiment of the present invention may be a kit comprising a drug.

"Combination" means the combined application of active ingredients. For example, the combination of the first active ingredient and the second active ingredient includes "a method of administering as a single preparation containing the first active ingredient and the second active ingredient" (i.e., a method of administering the first active ingredient and the second active ingredient as a compound (compound medicine)), and "a method of administering the first active ingredient and the second active ingredient as separate preparations simultaneously or separately". In the latter method, the preparation containing the first active ingredient can be administered first, or the preparation containing the second active ingredient can be administered first. The latter method can be any of "a method of preparing the first active ingredient and the second active ingredient separately and administering them simultaneously by the same administration route", "a method of preparing the first active ingredient and the second active ingredient separately and administering them separately by the same administration route with a time difference", "a method of preparing the first active ingredient and the second active ingredient separately and administering them simultaneously by different administration routes (administered from different parts of the same patient)", and "a method of preparing the first active ingredient and the second active ingredient separately and administering them separately by different administration routes with a time difference". In the case of "the first active ingredient and the second active ingredient are formulated separately and administered simultaneously via the same administration route", the two agents (the agent containing the first active ingredient and the agent containing the second active ingredient) may be mixed immediately before administration. "Separately" means that one agent is administered before or after the other agent is administered.

In other words, "combination use" can also be referred to as a method of using the drug in a state where one active ingredient is present in the patient's body, and another active ingredient is present in the patient's body. That is, it is preferred that the drug is administered in such a manner that the first active ingredient and the second active ingredient are simultaneously present in the patient's body, for example, in the blood, and it is preferred that a certain drug containing the first active ingredient and the second active ingredient are administered to the patient simultaneously, or another drug containing the other active ingredient is administered within 48 hours after administration of a certain drug containing the first active ingredient or the second active ingredient.

Compound 1 or its salt or solvate thereof may be Compound 1, may be a solvate of Compound 1, or may be a hydrate of Compound 1.

As the salt of compound 1, the salts allowed by pharmacy are listed, specifically, salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; salts with organic acids such as acetic acid, succinic acid, fumaric acid, maleic acid, tartaric acid, citric acid, lactic acid, stearic acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, etc.; salts with alkali metals such as sodium and potassium; salts with alkaline earth metals such as calcium and magnesium; ammonium salts; salts with amino acids such as arginine, etc. These salts are prepared, for example, by contact of the compound with an acid or a base. In addition, in this specification, the solvate is not particularly limited as long as it forms a single molecular group with a solvent, and is a solvate formed by a solvent that can be taken (ingested) with the administration of the medicament. For example, not only hydrates, alcohol hydrates (ethanol hydrate, methanol hydrate, 1-propanol hydrate, 2-propanol hydrate, etc.), solvates with a single solvent such as dimethyl sulfoxide can be listed, but also solvates formed with multiple molecules of solvents for one molecule of the compound, and solvates formed with multiple solvents for one molecule of the compound can be listed. In the case where the solvent is water, it is called a hydrate. Solvates are prepared, for example, by contacting a compound with a solvent, and solvates of salts are prepared, for example, by contacting a salt of a compound with a solvent. The salt of a compound or the compound or the solvate of the salt of the compound is the same as the compound (free body) as an active body. In the present specification, "(their) solvate" includes a solvate of compound 1, a solvate of a molecular targeting agent, a solvate of a salt of compound 1, and a solvate of a salt of a molecular targeting agent.

Compound 1 or its salt or its solvate may have crystalline polymorphism, and may be a single substance or a mixture of any crystalline forms. Compound 1 or its salt or its solvate may also include amorphous bodies.

The dosage of compound 1 or its salt or their solvate is preferably 0.001 to 100 mg (0.001 to 100 mg/kg/day) per 1 kg of the body weight of the administered subject, more preferably 0.003 to 20 mg/kg/day, and more preferably 0.003 to 10 mg/kg/day. When the dosage of compound 1 is within these ranges, the therapeutic or preventive effect of cancer is higher. The number of administrations of compound 1, for example, can be once a day, twice a day, or three times a day, more preferably once a day. The same dosage and/or usage can also be applied to molecular targeted agents.

Compound 1 can be prepared, for example, according to the method described in International Publication No. 2021/090855, by peptide synthesis using the Fmoc method, or by Solid phase peptide synthesis (published by Bachem, [retrieved on December 24, 2021], Internet URL: https://www.bachem.com/wp-admin/admin-ajax.php?juwpfisadmin=false&action=wpfd&task=file.download&wpfd_category_id=180&wpfd_file_id=213455&token=&preview=1, or Peptide Guide, No: 2004505, Global Marketing BachemAG, 2020.).

In this specification, "molecular targeting agent" means an agent having an anti-tumor effect, which specifically inhibits a specific protein. In addition, in this specification, compound 1 or its salt or their solvate is not included in the molecular targeting agent. As a molecular targeting agent, MAPK/ERK pathway inhibitors are listed, which can be used as a drug for the treatment or prevention of cancer. In this specification, "MAPK/ERK pathway inhibitor" refers to an inhibitor of a target involved in MAPK/ERK signaling, including inhibitors of growth factors and their receptors, and kinase inhibitors. For example, as MAPK/ERK pathway inhibitors, EGFR inhibitors, VEGF inhibitors, SHP2 inhibitors, KRAS inhibitors, NRAS inhibitors, HRAS inhibitors, KRAS-GTP inhibitors, NRAS-GTP inhibitors, HRAS-GTP inhibitors, KRAS-G12C inhibitors, ERK inhibitors, AKT inhibitors, MEK inhibitors can be listed. The molecular targeting agent may be at least one selected from EGFR inhibitors, VEGF inhibitors, SHP2 inhibitors, KRAS-G12C inhibitors and MEK inhibitors, and may be EGFR inhibitors, VEGF inhibitors, SHP2 inhibitors, KRAS-G12C inhibitors or MEK inhibitors. Preferably, the molecular targeting agent is an EGFR inhibitor. The EGFR inhibitor may be at least one selected from gefitinib, erlotinib, afatinib, osimertinib, lapatinib or their salts or their solvates, and anti-EGFR antibodies, and may be an anti-EGFR antibody. Preferably, the EGFR inhibitor is an anti-EGFR antibody. The anti-EGFR antibody may have a heavy chain variable region comprising the amino acid sequence of sequence number 1 and a light chain variable region comprising the amino acid sequence of sequence number 2, and may be cetuximab. More preferably, the EGFR inhibitor is cetuximab. The VEGF inhibitor may be at least one selected from sorafenib, pazopanib, sunitinib, axitinib, regorafenib or their salts or solvates, and anti-VEGF antibodies, may be anti-VEGF antibodies, may have a heavy chain variable region comprising the amino acid sequence of sequence number 3, and a light chain variable region comprising the amino acid sequence of sequence number 4, may be bevacizumab, ramucirumab, or aflibercept. As a VEGF inhibitor, an anti-VEGF antibody is preferred, and bevacizumab is more preferred. The SHP2 inhibitor may be at least one selected from RMC4550, TNO155, RLY1971, SHP099, NSC-87877 and their salts and solvates thereof, may be RMC4550, TNO155 and their salts and solvates thereof. Preferably, as a SHP2 inhibitor, one selected from RMC4550, TNO155, RLY1971 and their salts and solvates thereof may be cited. More preferably, as SHP2 inhibitors, RMC4550, TNO155 or their salts or their solvates can be listed. KRAS-G12C inhibitors can be selected from at least one of sotolasib, MRTX849 and their salts and their solvates, preferably, sotolasib or its salts or their solvates, or MRTX849 or its salts or their solvates. MEK inhibitors can be selected from at least one of the following compounds represented by formula (2), trametinib, selumetinib, CH4987655 and their salts and their solvates, preferably, the following compounds represented by formula (2), trametinib or their salts or their solvates. More preferably, as MEK inhibitors, the following compounds represented by formula (2) or their salts or their solvates can be listed.

[Chemical formula 5]

Molecular targeting agents can be obtained from commercial suppliers or prepared according to known methods.

Gefitinib (CAS No.: 184475-35-2, N-(3-chloro-4-fluoro-phenyl)-7-methoxy-6-(3-morpholin-4-ylpropoxy)quinazolin-4-amine) is a compound represented by the following formula (3). Gefitinib is used as Iressa.

[Chemical formula 6]

Erlotinib (CAS No.: 183321-74-6, N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine) is a compound represented by the following formula (4). Erlotinib is used as erlotinib hydrochloride or Tarceva.

[Chemical formula 7]

Afatinib (CAS No.: 439081-18-2, N-[4-[(3-chloro-4-fluorophenyl)amino]-7-[[(3S)-tetrahydro-3-furanyl]oxy]-6-quinazolinyl]-4(dimethylamino)-2-butenamide is a compound represented by the following formula (5). Afatinib is used as afatinib maleate or Gitari.

[Chemical formula 8]

Osimertinib (CAS No.: 1421373-65-0, N-(2-{2-dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide is a compound represented by the following formula (6). Osimertinib is used as osimertinib mesylate or Tagrisso.

[Chemical formula 9]

Lapatinib (CAS No.: 231277-92-2, N-[3-chloro-4-[(3-fluorophenyl)methoxy]phenyl]-6-[5-[(2-methylsulfonylethylamino)methyl]-2-furyl]quinazolin-4-amine) is a compound represented by the following formula (7). Lapatinib is used as lapatinib tosylate hydrate or tadalafil.

[Chemical formula 10]

RMC-4550 (CAS No.: 2172651-73-7, 3-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]dec-8-yl]-6-(2,3-dichlorophenyl)-5-methyl-2-pyrazinemethanol) is a compound represented by the following formula (8).

[Chemical formula 11]

TNO155 (CAS No.: 1801765-04-7, (3S, 4S)-8-(6-amino-5-((2-amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine) is a compound represented by the following formula (9).

[Chemical formula 12]

RLY1971 is a compound represented by CAS No.: 2668298-71-1.

SHP099 (CAS No.: 1801747-42-1, 6-(4-amino-4-methyl-1-piperidinyl)-3-(2,3-dichlorophenyl)-2-pyrazinamine) is a compound represented by the following formula (10).

[Chemical formula 13]

NSC-87877 (CAS No.: 56990-57-9, 8-hydroxy-7-[2-(6-sulfo-2-naphthyl)diazenyl]-5-quinolinesulfonic acid) is a compound represented by the following formula (11).

[Chemical formula 14]

Sotolacib (CAS No.: 2296729-00-3, 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one is a compound represented by the following formula (12). Sotolacib is used as AMG510 or Lumakras.

[Chemical formula 15]

MRTX849 (CAS No.: 2326521-71-3, 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile is a compound represented by the following formula (13). MRTX849 is used as adagracib.

[Chemical formula 16]

Trametinib (CAS No.: 871700-17-3, N-[3-[3-cyclopropyl-5-(2-fluoro-4-iodo-phenylamino)-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydro-2H-pyrido[4,3-d]pyrimidin-1-yl]phenyl]acetamide is a compound represented by the following formula (14). Trametinib is used as trametinib dimethyl sulfoxide adduct, or Mekinist.

[Chemical formula 17]

Compound 2 (CAS No.: 2677856-11-8, 2-(4-cyclopropyl-2-fluoro-anilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)-4-pyridyl]methyl]benzamide, 2-(4-cyclopropyl-2-fluoro-anilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)-4-pyridyl]methyl]benzamide) is a compound represented by the following formula (2). Compound 2 is used, for example, as a sodium salt.

[Chemical formula 18]

CH4987655 (CAS No.: 874101-00-5, 3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-N-(2-hydroxyethoxy)-5-[(tetrahydro-3-oxo-2H-1,2-oxazin-2-yl)methyl]benzamide, RO4987655) is a compound represented by the following formula (15).

[Chemical formula 19]

Selumetinib (CAS No.: 606143-52-6, 6-(4-bromo-2-chloroanilino)-7-fluoro-N-(2-hydroxyethoxy)-3-methylbenzimidazole-5-carboxamide) is a compound represented by the following formula (16). Selumetinib is used as selumetinib sulfate or KOSELUGO.

[Chemical formula 20]

Sorafenib (CAS No.: 284461-73-0, 4-[4-[[4-chloro-3-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-N-methyl-pyridine-2-carboxamide) is a compound represented by the following formula (17). Sorafenib is used as sorafenib tosylate or Nexavar.

[Chemical formula 21]

Pazopanib (CAS No.: 444731-52-6, 5-[[4-[(2,3-dimethylindazol-6-yl)-methyl-amino]pyrimidin-2-yl]amino]-2-methyl-benzenesulfonamide) is a compound represented by the following formula (18). Pazopanib is used as pazopanib hydrochloride or Votrient.

[Chemical formula 22]

Sunitinib (CAS No.: 557795-19-4, N-[2-(diethylamino)ethyl]-5-[(Z)-(5-fluoro-2-oxo-dihydroindole-3-ylidene)methyl]-2,4-dimethyl-1H-pyrrole-3-carboxamide) is a compound represented by the following formula (19). Sunitinib is used as sunitinib malate or sutent.

[Chemical formula 23]

Axitinib (CAS No.: 319460-85-0, N-methyl-2-[[3-[(E)-2-(2-pyridyl)vinyl]-1H-indazol-6-yl]sulfanyl]benzamide is a compound represented by the following formula (20). Axitinib is used as Inlyta.

[Chemical formula 24]

Regorafenib (CAS No.: 755037-03-7, 4-[4-[[4-chloro-3-(trifluoromethyl)phenyl]carbamoylamino]-3-fluoro-phenoxy]-N-methyl-pyridine-2-carboxamide) is a compound represented by the following formula (21). Regorafenib is used as regorafenib hydrate or stivargo.

[Chemical formula 25]

The dosage of the molecular targeting agent is preferably 0.005 to 100 mg per kg of the body weight of the subject to be administered (0.005 to 100 mg/kg/day), more preferably 0.01 to 75 mg/kg/day, more preferably 0.02 to 50 mg/kg/day, and most preferably 5 to 40 mg/kg/day. When the dosage of the molecular targeting agent is within these ranges, the therapeutic or preventive effect of cancer is higher. The number of administrations of the molecular targeting agent can be, for example, more than once a week, or twice a week. Compound 1 or its salt or its solvate can also be applied with the same dosage and/or usage.

The dosage of compound 1 or its salt or its solvate and the molecular targeting agent is preferably 0.001-100 mg/kg/day for compound 1 or its salt or its solvate and 0.005-100 mg/kg/day for the molecular targeting agent per 1 kg of the body weight of the administered subject, more preferably 0.003-20 mg/kg/day for compound 1 or its salt or its solvate and 0.01-75 mg/kg/day for the molecular targeting agent, and more preferably 0.003-10 mg/kg/day for compound 1 or its salt or its solvate and 0.02-30 mg/kg/day for the molecular targeting agent. When the dosage of compound 1 or its salt or its solvate and the molecular targeting agent is within these ranges, the therapeutic or preventive effect of cancer is higher. The blood drug concentration such as AUC and Cmax and the patient's state can be observed, and the dosage and usage can be appropriately adjusted. The appropriate dosage and number of administrations (frequency of administration) can also refer to, for example, the appendix of the molecular targeted drug. The administration of Compound 1 or its salt or solvate thereof may be, for example, once or more a day, or may be twice a day. The administration of Compound 1 or its salt or solvate thereof is preferably once a day.

In the present invention, as the method of administration, oral, rectal, parenteral (intravenous, intramuscular, subcutaneous, percutaneous absorption), groove, vagina, intraperitoneal, intravesical or local (injection, drip, powder, ointment, gel or cream) administration and inhalation (oral or nasal spray) etc. are listed. As its mode of administration, for example, tablets, capsules, granules, powders, pills, aqueous and non-aqueous oral solutions and suspensions and parenteral solutions filled in containers suitable for being subdivided into each administration amount are listed. In addition, the mode of administration can also adapt to various methods of administration including the regulated release preparations such as subcutaneous implantation. The method of administration can be equally applied to the first active ingredient and the second active ingredient. The first active ingredient and the second active ingredient can be used as medicaments and can be used as preparations or compounding agents.

The first active ingredient may be, for example, any of the above-mentioned administration methods, and the second active ingredient may be, for example, an administration method that is the same as or different from the administration method of the first active ingredient. The interval between the administration of the first active ingredient and the second active ingredient may be, for example, an interval of 0 to 14 days, an interval of 0 to 10 days, or an interval of 0 to 7 days. The interval of administration may be determined, for example, by AUC, Cmax, Tmax, disappearance half-life, and the health status of the subject. For example, compound 1 or its salt or its solvate may be administered orally (tablets, capsules, etc.) and the molecular targeting agent may be administered by drip, respectively. For example, compound 1 or its salt or its solvate and the molecular targeting agent may be administered orally (tablets, capsules, etc.). For example, compound 1 or its salt or its solvate may be administered by drip and the molecular targeting agent may be administered orally (tablets, capsules, etc.), respectively. For example, compound 1 or its salt or its solvate and the molecular targeting agent may be administered by drip, respectively. In such a case, compound 1 or its salt or its solvate and the molecular targeting agent can be administered at any interval such as twice a day, once a day, once a week, once every two weeks, etc. More specifically, compound 1 or its salt or its solvate and the molecular targeting agent can be administered once a day, in which case, they can be administered before, between, or after a meal. Before, between, or after a meal can be before, between, or after any of breakfast, lunch, dinner, supper, or snacks. As long as the administration interval of compound 1 or its salt or its solvate and the molecular targeting agent is within 24 hours, both can be administered once a day. As needed, compound 1 or its salt or their solvate and molecular targeting agent are sometimes administered twice a day and once a day, once a day and once a day, once a day and twice a day, once a day and once a day, once a day and once a day, once a day and once a day, once a day and once a day, once a day and once a day, once a day and once a day, once a day and once a day, once a day and once a day, once a day and once a day, and twice a day and once a day. As needed, according to the AUC, Cmax, Tmax or disappearance half-life of compound 1 and/or the molecular targeting agent, or the health status of the patient, only the administration interval of the molecular targeting agent can be increased or decreased, or only the administration interval of compound 1 or its salt or their solvate can be increased or decreased. The molecular targeting agent can be started on the day when compound 1 or its salt or their solvate is started, and the day when compound 1 or its salt or their solvate is started can be different from the day when the molecular targeting agent is started. The administration period can be 7 days as one course of treatment, the total number of courses of treatment can be more than 1 course of treatment, or more than 2 courses of treatment. In addition, each course of treatment can be carried out continuously, or a drug-free period can be set. A drug-free period may also be provided in the middle of the course of treatment. As required, only Compound 1 or its salt or its solvate may be continuously administered during the course of treatment while the molecular targeting agent may be discontinued, or only Compound 1 or its salt or its solvate may be discontinued while the molecular targeting agent may be continued. Compound 1 or its salt or its solvate and the molecular targeting agent may also be included in the same tablet, capsule, etc.

The pharmaceutical preparations of the present invention can be formulated by known methods in addition to active ingredients such as compound 1 or its salt, or their solvates and molecular targeting agents, etc., by introducing drug-acceptable carriers. Excipients, adhesives, lubricants, colorants, fragrances and stabilizers, emulsifiers, absorption promoters, surfactants, pH regulators, preservatives, antioxidants, etc. commonly used in formulation can be used, and ingredients commonly used as raw materials for pharmaceutical preparations can be formulated by conventional methods. In the preparation, the active ingredients used in the medicine can be processed into the best shape and properties, i.e., dosage form, according to its method of use and purpose of use by known methods. Commonly used dosage forms include liquid pharmaceutical preparations (liquid preparations) such as injections, suspensions, emulsions, eye drops, and solid pharmaceutical preparations (solid preparations) such as tablets, powders, granules, granules, coated tablets, capsules, dry syrups, lozenges, suppositories, etc., but are not limited to these. For example, a preparation containing the active ingredients exemplified in compound 1 or its salt, or their solvates, and molecular targeting agents can be used as the medicine of the present invention.

The therapeutic or preventive medicine of the present invention can be used by mixing any one or both of the first active ingredient and the second active ingredient with a drug-permissible additive. As a drug-permissible additive, a person skilled in the art can appropriately select a known additive, and a plurality of additives can be appropriately used as needed. The additive can be appropriately selected according to an oral agent or an injection, for example, mixed with water, ethanol, saline, liquid paraffin, a surfactant, sucrose, etc., and the resulting mixture can be encapsulated.

As cancer, solid cancer or blood cancer are listed, and any type is composed of abnormal cells that proliferate uncontrollably. Solid cancer forms one or more tumors, and blood cancer circulates throughout the body via the bloodstream. Cancer can be, for example, at least one selected from lung cancer, esophageal cancer, gastric cancer, colorectal cancer, uterine cancer, ovarian cancer, pancreatic cancer, bladder cancer, thyroid cancer, skin cancer, leukemia, malignant lymphoma and multiple myeloma. Cancer can be a cancer related to abnormalities in the RAS gene. Preferably, the cancer is lung cancer, pancreatic cancer or colorectal cancer.

The term "abnormality" as used herein refers to a state in which various chromosomes, genes, proteins, and signal transduction cascades in cells deviate from normal operation or control, and usually, their constant activation becomes the cause.

"Abnormal activation" in this specification refers to a state in which various chromosomes, genes, proteins and signal cascades in cells are more activated than normal cells, for example, a case where a gene mutation is one of the causes (Cancer Metastasis Rev, 2013, 32, 147-162). In order to directly quantitatively measure the involvement of mutants in cells, for example, cell activation can be measured by LC/MS-MS-based analysis (Cancer Discov., 2016, 6, 316-329).

"Protein amplification" refers to the amplification of proteins involved in the MAPK/ERK pathway (e.g., KRAS, NRAS, HRAS, KRAS-GTP, NRAS-GTP, HRAS-GTP, ERK, AKT, MEK, pERK, pAKT, pMEK), etc., and the synthesis and amplification of proteins in living cells. Protein amplification can be measured by Western blotting, comparative genomic hybridization (CGH), etc., and confirmed by comparison with proteins in normal cells (Nat. Rev. Drug Discov., 2020, 19, 533-552, Molecular Cytogenetics, 2015, 8: 103).

In one aspect, cancer includes cancer associated with abnormalities in RAS genes. Cancer associated with abnormalities in RAS genes refers to, for example, cancer cells having mutations in the coding region of RAS genes and/or amplification of the number of copies of RAS genes, and cancers in which an increase in RAS activity (including an increase in the production of RAS proteins) is found. Mutations in the coding region of RAS genes refer to, for example, deletions, additions, or substitutions of one or more bases in the coding region of RAS genes. Amplification of the number of copies of RAS genes refers to the presence of RAS genes with more copies than the number of copies of RAS genes that are usually present on chromosomes. More copy numbers mean more than normal cells, and as long as the number of copies is more than 4, it can be called copy number amplification.

Whether a cancer is a cancer related to abnormality of the RAS gene can be determined by confirming whether a subject suffering from cancer has abnormality of the RAS gene. For example, by identifying a mutation in the base sequence of the coding region of the RAS gene in a biological sample obtained from a subject suffering from cancer, and further confirming that the mutated RAS protein is expressed, it can be determined that the cancer is a cancer related to abnormality of the RAS gene.

Whether the cancer is a cancer related to the abnormality of the RAS gene can be determined by confirming whether the object with cancer has an amplification of the copy number of the RAS gene. For example, by confirming that the copy number of the RAS gene in the biological sample obtained from the object with cancer is amplified, and/or the generation of the RAS protein is increased, it can be determined that the cancer is a cancer related to the abnormality of the RAS gene. The increase in the generation of the RAS protein can also be referred to as the overexpression of the RAS protein. Whether the generation of the RAS protein is increased can be confirmed, for example, by comparing the expression of the RAS protein with the expression of the usual RAS protein, the expression of the RAS protein in the object not suffering from the cancer, or the expression of the RAS protein in the object suffering from precancerous.

When a subject suffering from cancer has a mutation in the base sequence of the coding region of the RAS gene and the copy number of the RAS gene is amplified, it can be said that the cancer is related to the abnormality of the RAS gene.

Any method can be applied as long as it is a method capable of detecting a mutation in the coding region of the RAS gene in a biological sample of a subject suffering from cancer or amplification of the number of copies of the RAS gene, and can be performed by a person skilled in the art in a common manner. For example, the RAS gene can be isolated from a biological sample obtained from a subject suffering from cancer, amplified by PCR or the like, and its base sequence can be directly determined by a sequencer or the like. In addition, for example, a commercially available in vitro diagnostic drug capable of detecting a mutation in the RAS gene can be applied.

As isoforms of RAS protein, three are known: KRAS protein, NRAS protein, and HRAS protein. In one aspect, the RAS gene is one or more RAS genes selected from the group consisting of KRAS gene, NRAS gene, and HRAS gene. Preferably, the RAS gene is the KRAS gene.

In one aspect, cancer includes cancers associated with the production of mutant RAS proteins and/or increased production of RAS proteins. The production of mutant RAS proteins and/or increased production of RAS proteins are usually caused by abnormalities in the above-mentioned RAS genes.

In one aspect, the mutant RAS protein is at least one mutant RAS protein selected from a mutant KRAS protein, a mutant NRAS protein, and a mutant HRAS protein. Preferably, the mutant RAS protein is a mutant KRAS protein.

In one aspect, the mutant RAS protein has a mutation in at least one amino acid position selected from G12, G13 and Q61 in the amino acid sequence of a wild-type human KRAS protein, the amino acid sequence of a wild-type human NRAS protein, or the amino acid sequence of a wild-type human HRAS protein. In any of the amino acid sequences of wild-type human KRAS proteins, NRAS proteins and HRAS proteins, the 12th and 13th amino acids are glycine, and the 61st amino acid is glutamine.

In one aspect, the mutant RAS protein is a mutant KRAS protein and has at least one amino acid mutation selected from G12A, G12C, G12D, G12S, G12V, G13D, Q61H, and Q61K compared to the amino acid sequence of wild-type KRAS.

In one aspect, the mutant RAS protein is a mutant NRAS protein and has at least one amino acid mutation selected from the group consisting of G12C, G12D, G13D, G13V, Q61K, and Q61L compared to the amino acid sequence of wild-type NRAS.

In one aspect, the mutant RAS protein is a mutant HRAS protein and has an amino acid mutation of G13R compared to the amino acid sequence of wild-type HRAS.

Example

The following examples are used to further illustrate the content of the present invention, but the present invention is not limited to the content. All compounds and reagents were obtained from commercial suppliers or synthesized using known methods. In this example, the molecular targeting agent is sometimes referred to as a combination compound.

Compound 1 ((5S,8S,11S,15S,18S,23aS,29S,35S,37aS)-8-((S)-sec-butyl)-18-cyclopentyl-29-(3,5-difluoro-4-(trifluoromethyl)phenethyl)-36-ethyl-11-isobutyl-N,N,5,6,12,16,19,33-octamethyl-35-(4-methylbenzyl)-4,7,10,13,17,20,23,28,3 1,34,37-Undecanoic acid tritetrahydro-2H,4H-spiro[azetidin-(azeto)[2,1-u]pyrrolo[2,1-i][1,4,7,10,13,16,19,22,25,28,31]undecazacyclotriacontahedenadecene-21,1'-cyclopentane]-15-carboxamide) was prepared by the peptide synthesis method using the Fmoc method described in International Publication No. 2021/090855.

That is, it is prepared in the following 4 steps:

1) The carboxyl group of (3S)-4-(dimethylamino)-3-[9H-fluoren-9-ylmethoxycarbonyl(methyl)amino]-4-oxo-butyric acid was loaded onto 2-chlorotrityl resin, and a peptide extension reaction was performed from the N-terminus of the amino acid by the Fmoc method corresponding to the sequence of compound 1.

2) Cleavage reaction of peptide from 2-chlorotrityl resin

3) Amide cyclization reaction caused by condensation of the carboxyl group released from the 2-chlorotrityl resin by the cleavage reaction with the amino group at the N-terminus of the peptide chain (N-methylleucine)

4) Purification of compounds by preparative HPLC.

Compound 2 (2-(4-cyclopropyl-2-fluoro-anilino)-3,4-difluoro-5-[[3-fluoro-2-(methylsulfamoylamino)-4-pyridinyl]methyl]benzamide) was prepared according to the method described in International Publication No. 2021/149776.

Right now,

1) 2,3,4-trifluorobenzoic acid available from a commercial supplier is reacted with iodine to give 2,3,4-trifluoro-5-iodo-benzoic acid.

2) The compound obtained in the above reaction is subjected to a coupling reaction with potassium vinyl trifluoroborate in the presence of a palladium catalyst to obtain 2,3,4-trifluoro-5-vinyl-benzoic acid.

3) The compound obtained in the above reaction is coupled with 2-fluoro-4-iodo-aniline to obtain 3,4-difluoro-2-(2-fluoro-4-iodo-anilino)-5-vinyl-benzoic acid.

4) The compound obtained in the above reaction is treated with sodium periodate and osmium 4 oxide to obtain 3,4-difluoro-2-(2-fluoro-4-iodo-anilino)-5-formyl-benzoic acid.

5) The compound obtained in the above reaction was reacted with a hexane solution of diazomethyltrimethylsilane in methanol to obtain methyl 3,4-difluoro-2-(2-fluoro-4-iodoanilino)-5-formylbenzoate.

6) The compound obtained in the above reaction is reacted with 4-methylbenzenesulfonylhydrazide to obtain methyl 3,4-difluoro-2-(2-fluoro-4-iodoanilino)-5-[(E)-[(4-methylphenyl)sulfonylhydrazinylidene]methyl]benzoate.

7) The compound obtained in the above reaction is reacted with [2-[(2,4-dimethoxyphenyl)methylamino]-3-fluoropyridin-4-yl]boric acid to obtain methyl 5-[[2-[(2,4-dimethoxyphenyl)methylamino]-3-fluoropyridin-4-yl]methyl]-3,4-difluoro-2-(2-fluoro-4-iodoanilino)benzoate.

8) The compound obtained in the above reaction is reacted with trifluoroacetic acid to obtain methyl 5-[(2-amino-3-fluoropyridin-4-yl)methyl]-3,4-difluoro-2-(2-fluoro-4-iodoanilino)benzoate.

9) The compound obtained in the above reaction was hydrolyzed with lithium hydroxide for esterification and then treated with hydrochloric acid to obtain 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzoic acid hydrochloride.

10) The compound obtained in the above reaction is reacted with EDC·HCl, HOOBt, and ammonia methanol solution to obtain 5-((2-amino-3-fluoropyridin-4-yl)methyl)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide. Subsequently, cyclopropylzinc bromide is reacted to obtain 5-((2-amino-3-fluoropyridin-4-yl)methyl)-2-((4-cyclopropyl-2-fluorophenyl)amino)-3,4-difluorobenzamide.

11) The compound obtained in the above reaction (2.47 g, 5.74 mmol) was dissolved in anhydrous DMF (28.7 mL), pyridine (2.78 mL, 34.4 mmol) and 4-nitrophenyl methylaminosulfonate (4.00 g, 17.2 mmol) were added, and stirred at 40°C for 2.5 hours. The reaction mixture was cooled to room temperature and water (24.7 mL) was added. Acetonitrile (3 mL) and water (19.8 mL) were added, and after stirring for 10 minutes, the solid was filtered. The obtained solid was washed with water/acetonitrile (1/1, 49.4 mL) to obtain compound 2 (2.56 g, 85%) as a colorless solid. Molecular targeting agents other than compound 2 (sotoracib, RMC-4550, trametinib and cetuximab) were provided by commercial suppliers or manufacturers.

＜Pharmacological Test Examples＞

(Example 1 Evaluation of RAS signaling inhibitory activity in vitro)

The effects of compound 1, compound 2, RMC-4550, trametinib and sotolacib, and the combination of these compounds on the phosphorylation of ERK, AKT and MEK in human cancer cells, and the binding of KRAS, NRAS and HRAS to GTP were studied by the following protein blotting method. Human cancer cell lines were inoculated on a 3D culture plate PrimeSurface (Sumitomo Bakelite) and cultured at 37°C for 24 hours. Thereafter, compound 1 and a combined compound (molecular targeting agent) were added to the cells. Cultured at 37°C in a 5% carbon dioxide incubator, the cells were recovered at the time point after the treatment time shown in Table 1. The name of the human cancer cell line, culture medium, number of cells inoculated, concentration of compound 1, combined compound, concentration of combined compound and treatment time are shown in Table 1. The recovered cells were dissolved with a lysis buffer (Cell Signaling Technology) containing a protease inhibitor (Sigma-Aldrich) and a phosphatase inhibitor (Sigma-Aldrich). The protein concentration was measured using a BCA protein assay kit (ThermoFisher Scientific), and the protein solution 1 was adjusted to 10 to 20 μg per sample.

From 300 to 1000 μg of the obtained protein solution 1, RAS-GTP as an activated form was recovered by Active Ras Pull-Down and Detection Kit (ThermoFisher Scientific) to adjust protein solution 2. SDS-PAGE was performed using the above protein solutions 1 and 2 and a gradient gel with an acrylamide concentration of 5-20%, and the migrating protein was transferred to a PVDF membrane by Trans-Blot Turbo Transfer System (Bio-Rad Laboratories). After blocking, the PVDF membrane was reacted with the primary antibody and the secondary antibody in this order. The bands were detected by Chemi-lumi One Super (Nakarai Tesk) chemiluminescence and ChemiDoc Touch Imaging System (Bio-Rad Laboratories). The primary antibody, secondary antibody, dilution concentration of each antibody, reaction temperature and reaction time used are shown in Tables 2 and 3. The results of the electrophoretic diagrams showing the results of the protein blotting for the evaluation of the RAS signal inhibitory activity are shown in Figures 1 to 6. "pERK", "pAKT" and "pMEK" represent phosphorylated ERK, AKT and MEK, respectively. "KRAS-GTP", "NRAS-GTP" and "HRAS-GTP" represent KRAS, NRAS and HRAS bound to GTP, respectively. In Figures 1 to 6, when compound 1 and a combined compound (molecular targeted agent) are used in combination, the inhibitory effect of RAS signaling is enhanced compared to the case where compound 1 or a combined compound (molecular targeted agent) is used as a single agent.

[Table 1]

[Table 2]

[Table 3]

(Example 2 Evaluation of cell proliferation inhibitory activity of compound 1 and combined compound (molecular targeting agent))

In the ultra-low adhesion surface U-bottom 384 plate, human cancer cell lines were inoculated at a concentration of 750 cells per well and cultured at 37°C in a 5% carbon dioxide incubator. After 24 hours, CellTitorGlo2.0 (Promega) was added to a portion of the wells (3 wells), and the ATP level ( _V0 ) was determined using Envision plate readers (Perkin Elmer). _V0 was taken as the ATP level at 0 hours. In each of the other 3 wells, DMSO alone, or compound 1 and/or the combination compound were added. The highest concentration of compound 1 was 300 nM, and the highest concentration of the combination compound was 10 or 3000 nM, and a series of 8-stage dilutions with a common ratio of 3 was used. In Table 4, for example, "0.14-300" means the case where the maximum concentration is "300" nM, indicating the application of dilutions of "300, about 100, about 33, about 11, about 3.7, about 1.2, about 0.41 or about 0.14 nM". After culturing for 144 hours after adding only DMSO, or compound 1 and/or a combination compound, the ATP levels (T and V) of cells to which only DMSO was added (DMSO-treated group) or cells to which compound 1 and/or a combination compound was added (compound 1 and/or a combination compound-treated group) were measured. The cancer cell line name, culture medium, compound 1 concentration, combination compound and combination compound concentration are shown in Table 4. The proliferation inhibition rate (GI%: growth inhibition%) was calculated using the following formula. As shown in Figures 7a, 8a and 9a, the concentration-dependent proliferation inhibition effect of compound 1 and the combination compound is shown.

(1-(TV ₀ )/V ₀ )×100 (in the case of T＜V ₀ )

(1-(TV ₀ )/(VV ₀ ))×100(in case of T≧V ₀ )

T: ATP signal value at 144 hours in the group treated with compound 1 and/or combination compounds

V: ATP signal value of DMSO-treated group at 144 hours

V ₀ : ATP signal value of the untreated group

The obtained GI values were used to perform an isobologram analysis according to the following procedure. The following GI values were used to evaluate the combined effect.

(1) Calculation of GI% (set GI%) for evaluating the combined effect of compound 1 and the combined compound, and the compound concentration that gives the set GI% (set GI concentration)

·Combination study of compound 1 and RMC-4550 (NCI-H358): As the set GI%, 155% (sometimes also expressed as "GI155%" or "GI155") was used (the set GI concentration of compound 1 was 0.033 μM, and the set GI concentration of RMC-4550 was 0.55 μM).

·Combination study of compound 1 and RMC-4550 (NCI-H441): As the set GI%, 100% (sometimes also expressed as "GI100%" or "GI100") was used (the set GI concentration of compound 1 was 0.0055 μM, and the set GI concentration of RMC-4550 was 0.35 μM).

· Combination study of Compound 1 and Trametinib (NCI-H358): As the set GI%, 50% (sometimes also expressed as "GI50%" or "GI50") was used (the set GI concentration of Compound 1 was 0.0046 μM, and the set GI concentration of Trametinib was 0.00035 μM).

(2) Combination study of the concentrations of compound 1 and the combination compound given at each set GI% in (1), and visualization of the combination effect by graph

The horizontal axis represents the value obtained by dividing the concentration of the combined compound by the set GI concentration, and the vertical axis represents the value obtained by dividing the concentration of the compound 1 by the set GI concentration.

The concentrations of the combination of Compound 1 and the combination compound administered at a set GI% in various cell lines were measured, and the values corresponding to the horizontal and vertical axes were plotted on the graph.

The results of the cell proliferation inhibitory activity evaluation of compound 1 and the combined compound are shown in Figures 7 to 9. Each of Figures 7b, 8b and 9b is a figure called an isobologram. The dotted line connecting the "1.0" of the vertical axis of the isobologram and the "1.0" of the horizontal axis indicates that the proliferation inhibitory effect of the case where compound 1 is combined with the combined compound is a simple total of the proliferation inhibitory effects of the case where each compound is applied alone (the case where the effects are added). The graph (plot) below the dotted line means that in the case where compound 1 is combined with the combined compound, a synergistic proliferation inhibitory effect is shown. The farther the graph is from the dotted line, the stronger the synergistic effect is shown. As shown in Figures 7b, 8b and 9b, since the graphs in each figure are far away from the dotted line, it is known that the combination of compound 1 and the combined compound shows a synergistic proliferation inhibitory effect.

[Table 4]

Cell Name Culture medium Compound 1 concentration (nM) Combined compounds Concentration of combined compound (nM) NCI-H358 RPMI-1640 + 10% serum 0.14～300 RMC-4550 1.4～3000 NCI-H441 RPMI-1640 + 10% serum 0.14～300 RMC-4550 1.4～3000 NCI-H358 RPMI-1640 + 10% serum 0.14～300 Trametinib 0.0046～10

(Example 3 Evaluation of in vivo antitumor activity by combination of Compound 1 and cetuximab)

The in vivo antitumor activity of the combination of compound 1 and cetuximab was evaluated using a xenograft model transplanted with the colorectal cancer cell line LoVo (KRAS-G13D). 5×10 ⁶ cells were transplanted subcutaneously in each nude mouse, and the nude mice were randomized when the tumor volume reached 200-300 mm ³ , and each 8 mice were divided into 4 groups. The single-dose administration group (8 mice per group) was administered with compound 1 or cetuximab, and the two-dose administration group (8 mice per group) was administered with compound 1 and cetuximab. The administration period was 10 days, compound 1 was orally administered once a day, and cetuximab was administered intravenously at a rhythm of twice a week. Compound 1 was administered at 10 mg/kg per time, and cetuximab was administered at 40 mg/kg per time. Compound 1 was dissolved in 10% DMSO/5% Cremphor EL/85% distilled water and administered. Cetuximab was diluted with physiological saline and administered. The tumor growth inhibition rate (TGI%: tumor growth inhibition%) was calculated using the following formula. The results of the in vivo antitumor activity evaluation are shown in Figure 10. Compared with the vehicle group, the increase in tumor volume in the drug administration group (compound 1 single-dose administration group, cetuximab single-dose administration group, and compound 1 and cetuximab double-dose administration group) was inhibited. Furthermore, in the comparison between the single-dose administration group and the double-dose administration group (combination group), the tumor volume in the double-dose administration group was reduced, indicating that the in vivo antitumor activity was enhanced by combining compound 1 and cetuximab.

Tumor proliferation inhibition rate (TGI%) = (1-TV/CV) × 100

Tumor volume (mm ³ ) = 1/2 × major diameter (mm) × minor diameter (mm) × minor diameter (mm)

Tumor volume change (mm ³ ) = tumor volume on day 32, 36 or 39 - tumor volume on day 29

TV: Average values of tumor volume changes in the compound 1 single-dose group, cetuximab single-dose group, or combination group

CV: Mean tumor volume change of the vehicle group

(Example 4 Evaluation of RAS signaling inhibitory activity in vivo)

After the completion of Example 3, compound 1 (10 mg/kg) and cetuximab (40 mg/kg) were further administered to each nude mouse (n=3), and the tumor was recovered 4 hours later and frozen in liquid nitrogen and stored at -80°C. Thereafter, the recovered tumor was disrupted with Multi-Beads Shocker (Yasui Instruments) under cooling with liquid nitrogen, and the RAS signal inhibitory activity by compound 1 and cetuximab in the xenograft model was evaluated based on the same protocol as in Example 1 (in vitro RAS signal inhibitory activity evaluation). The primary antibody, secondary antibody, dilution concentration of each antibody, reaction time and reaction temperature used are shown in Tables 5 and 6. The results of the electrophoresis diagram showing the results of the Western blotting for the in vivo RAS signal inhibitory activity evaluation are shown in Figure 11. "KRAS-GTP" means "KRAS" bound to GTP, and "pERK", "pAKT" and "pEGFR" respectively represent phosphorylated "ERK", "AKT" and "EGFR". Compared with the case where Compound 1 or cetuximab was used as a single agent, the case where Compound 1 and cetuximab were used in combination showed a decrease in KRAS-GTP and pERK, and enhanced inhibition of RAS signaling.

[Table 5]

[Table 6]

(Example 5 Evaluation of in vivo antitumor activity by combination of Compound 1 and bevacizumab)

The in vivo antitumor activity of the combination of compound 1 and bevacizumab was evaluated using a xenograft model transplanted with the pancreatic cancer line AsPC-1 (KRAS-G12D). 5×10 ⁶ cells were transplanted subcutaneously per nude mouse, and the nude mice were randomized when the tumor volume reached 200-300 mm ³ , and each 8 mice were divided into 4 groups. The single-dose administration group (8 mice per group) was administered with compound 1 or bevacizumab, and the two-dose administration group (8 mice per group) was administered with compound 1 and bevacizumab. The administration period was 17 days, compound 1 was orally administered once a day, and bevacizumab was administered by intraperitoneal injection at a rhythm of twice a week. Compound 1 was administered at 10 mg/kg per time, and bevacizumab was administered at 2.5 mg/kg per time. Compound 1 was dissolved in 10% DMSO/5% Cremphor EL/85% distilled water and administered. Bevacizumab was diluted with physiological saline and administered. The tumor proliferation inhibition rate (TGI%: tumor growth inhibition%) was calculated using the following formula. The results of the in vivo antitumor activity evaluation are shown in Figure 12. Compared with the vehicle group, the increase in tumor volume in the compound 1 single-dose administration group and the compound 1 and bevacizumab 2-dose administration group was suppressed. On the other hand, in the bevacizumab single-dose administration group, no significant inhibition of tumor volume increase was observed. Furthermore, in the comparison between the single-dose administration group and the 2-dose administration group (combination group), due to the reduction in tumor volume in the 2-dose administration group, the in vivo antitumor activity enhancement effect was shown by combining compound 1 with bevacizumab.

Tumor proliferation inhibition rate (TGI%) = (1-TV/CV) × 100

Tumor volume (mm ³ ) = 1/2 × major diameter (mm) × minor diameter (mm) × minor diameter (mm)

Tumor volume change (mm ³ ) = tumor volume after 25, 29, 32, 36, or 39 days - tumor volume on day 22

TV: the average value of tumor volume change in compound 1 group, bevacizumab group or combination group CV: the average value of tumor volume change in vehicle group

(Example 6 Evaluation of cell proliferation inhibitory activity by combination of compound 1 and sotolacib)

In a 96-well 2D culture plate (Corning), 10,000 cells per well were inoculated with human colorectal cancer cell line SW837 (n=3) and cultured at 37°C in a 5% carbon dioxide incubator. Leibovitz’s L-15 + 10% serum was used as the culture medium. 24 hours after inoculation, the culture medium was exchanged to contain DMSO/Compound 1 30nM/Sotolacib 300nM/Compound 1 30nM + Sotolacib 300nM, and the measurement of cell density every 24 hours was started by Icucyte ZOOM. About twice a week, the culture medium was exchanged for a culture medium containing the compound, and the measurement was carried out over a period of 52 days. The cell proliferation inhibitory activity of the combination of Compound 1 and Sotolacib is shown in Figure 13. In Figure 13, the vertical axis represents the proportion of cultured cells covering the adhesion surface of the culture vessel (cell density, cell occupancy rate, confluence), and the horizontal axis represents the number of days after the cell density is measured. As shown in FIG13 , the combination of Compound 1 and Sotolacib enhanced the proliferation inhibition effect in the human colorectal cancer cell line SW837.

(Example 7 Evaluation of cell proliferation inhibitory activity of compound 1 and combined compounds)

In a low-adhesion surface U-bottom 384 plate (Sumitomo Bakelite), 1000 cells (HCC-1171), 500 cells (NCI-H1373), 625 cells (NCI-H23), and 250 cells (UM-UC-3) were inoculated with human cancer cell lines (n=1) per well and cultured at 37°C in a 5% carbon dioxide incubator. When compound 1 was used in combination with MRTX849, a serial dilution method of 9 steps of 3 times the common ratio of compound 1 and 5 steps of 5 times the common ratio of MRTX849 was used. When compound 1 was used in combination with TNO155, a serial dilution method of 9 steps of 3 times the common ratio of compound 1 and 5 steps of 5 times the common ratio of TNO155 was used. The combination effect was verified by combining all the set concentrations of the two agents (including DMSO group, 10 concentrations and 6 concentrations). After adding compound 1 and/or the combination compound and culturing for 144 hours, the ATP amount of the compound treatment group was measured using CellTitorGlo2.0. The cancer cell line name, culture medium, compound name, and final test concentration of the compound are shown in Table 7. Based on the signal value obtained by CellTitorGlo2.0, the cell proliferation inhibition rate (CGI%: cell growth inhibition%) was calculated using the following formula.

Cell proliferation inhibition rate (CGI%) = (1-(TV-V0)/(CV-V0)) × 100

TV: ATP signal value at 144 hours in the compound-treated group

CV: ATP signal value of DMSO-treated group at 144 hours

V0: ATP signal value at 144 hours of culture medium only (no cell inoculation)

Using the obtained CGI values, analysis based on the isobologram method was carried out according to the following procedure.

The results of the evaluation of the cell proliferation inhibitory activity of compound 1 and the combination compound are shown in Figures 14 and 15. The X-axis (horizontal axis) and the Y-axis (vertical axis) are the concentrations of compound 1 and the combination compound, respectively. First, the concentration of a single dose of compound 1 and the concentration of a single dose of the combination compound required to exert the CGI50% value (HCC-1171, NCI-H1373, NCI-H23) or CGI70% value (UM-UC-3) are plotted on the X-axis or Y-axis. The straight line connecting the graph on the X-axis and the graph on the Y-axis indicates that the proliferation inhibitory effect of the case where compound 1 and the combination compound are used in combination is the simple sum of the proliferation inhibitory effects of the case where each compound is used alone (the case where the effects are added). Subsequently, when the drugs are used in combination, the concentrations of compound 1 and the combination compound required to exert their CGI values are plotted. Compared with the straight line connecting the graph on the X-axis and the graph on the Y-axis, when the graph when used in combination is located below, it means that the combination of the agents shows a synergistic proliferation inhibitory effect, and the farther the graph is from the straight line, the stronger the synergistic effect is shown. As shown in Figures 14 and 15, since the graphs in each figure are far from the straight line, it is known that the combination of compound 1 and the combined compound shows a synergistic proliferation inhibitory effect.

[Table 7]

(Example 8 Evaluation of in vivo antitumor activity by combination of Compound 1 and Sotolacib)

Nude mice transplanted with CTG-2579 (KRAS-G12C) as a lung cancer PDX (patient-derived xenograft) model were used to evaluate the in vivo antitumor activity of the combination of Compound 1 and Sotolacib. Tumor pieces were transplanted subcutaneously in nude mice, and randomization was performed when the tumor volume reached approximately 150-300 mm ³ , and administration of Compound 1 and/or the combination compound was started (n=8). Compound 1 and Sotolacib were orally administered once a day for 21 days. The dosage of the drug was 7.5 mg/kg for Compound 1 and 100 mg/kg for Sotolacib. Compound 1 was dissolved in 10% DMSO/5% Cremphor EL/85% distilled water and administered, and Sotolacib was dissolved in 10% DMSO/10% Cremphor EL/15% PEG400/2.64% HPBCD/62.36% distilled water and administered. According to the tumor volume data of the final measurement day, the following formula was applied to calculate the tumor proliferation inhibition rate (TGI%: tumor growth inhibition%). The results of the in vivo antitumor activity evaluation are shown in Figure 16. Compared with the vehicle group, the increase in tumor volume in the compound 1 single-dose administration group, the sotolacib single-dose administration group, and the compound 1 and sotolacib 2-dose administration group was inhibited. Further, in the comparison between the single-dose administration group and the 2-dose administration group (combination group), due to the reduction in tumor volume in the 2-dose administration group, the enhancement effect of in vivo antitumor activity was shown by combining compound 1 with sotolacib.

Tumor proliferation inhibition rate (TGI%) = (1-TV/CV) × 100

Tumor volume (mm ³ ) = 0.52 × long diameter (mm) × short diameter (mm) × short diameter (mm)

Tumor volume change (mm ³ ) = tumor volume after day 0 - tumor volume on day 0

TV: the average value of tumor volume change in compound 1 group, sotolacib group or combination group CV: the average value of tumor volume change in vehicle group

(Example 9 Evaluation of in vivo RAS signaling inhibitory activity by combination of Compound 1 and Sotolacib)

After Example 8, Compound 1 and Sotolacib were administered to nude mice (n=3) at 7.5 mg/kg and 100 mg/kg, respectively. After 4 hours, the tumor was recovered and frozen with liquid nitrogen and stored at -80°C. Thereafter, the recovered tumor (PDX sample) was broken with Multi-Beads Shocker (Yasui Instruments) under liquid nitrogen cooling, and the RAS signal inhibitory activity of Compound 1 and Sotolacib in PDX was evaluated based on the same protocol as the in vitro RAS signal inhibitory activity evaluation of Example 1. The primary antibody, secondary antibody, dilution concentration, reaction time, and reaction temperature used are shown in Tables 8 and 9. The results of the electrophoresis diagram showing the results of the protein blotting for the in vivo RAS signal inhibitory activity evaluation are shown in Figure 17. "KRAS-GTP" means "KRAS" bound to GTP, and "pERK" and "pAKT" mean phosphorylated "ERK" and "AKT", respectively. Compared with the case of single-dose application of compound 1 or sotolacib, the case of combined use of compound 1 and sotolacib showed a decrease in pERK and enhanced inhibition of MAPK signaling.

[Table 8]

[Table 9]

(Example 10 Evaluation of in vivo antitumor activity by combination of compound 1 and MRTX849)

The in vivo antitumor activity of the combination of compound 1 and MRTX849 was evaluated using a xenograft model transplanted with lung cancer line LU65 (KRAS-G12C) or bladder cancer line UM-UC-3 (KRAS-G12C). 5×10 ⁶ cells were transplanted subcutaneously per nude mouse. When the tumor volume reached 200-300 mm ³ , the nude mice were randomized and administration of compound 1 and/or the combination compound was started (n=6). Compound 1 and MRTX849 were orally administered once a day for 45 days. The dosage of the drug was 3 mg/kg for compound 1 and 100 mg/kg for MRTX849. Compound 1 was dissolved in 10% DMSO/5% Cremphor EL/85% distilled water and administered, and MRTX849 was dissolved in 50 mM Citrate buffer (pH5.0)/10% HPBCD and administered. According to the tumor volume data on the final measurement day, the tumor proliferation inhibition rate (TGI%: tumor growth inhibition%) was calculated using the following formula. The results of the in vivo antitumor activity evaluation are shown in Figure 18. In the lung cancer model (LU65) and the bladder cancer strain model (UM-UC-3), compared with the vehicle group, the increase in tumor volume in the MRTX849 single-dose administration group and the two-dose administration group of compound 1 and MRTX849 was inhibited. In the compound 1 single-dose administration group, no inhibition of the increase in tumor volume was observed. Regarding the MRTX849 single-dose administration group, the increase in tumor volume was inhibited immediately after the start of administration, but tumor re-growth was seen in multiple mice when the administration was continued. Since the two-dose administration group (combination group) long-term suppressed the re-growth of tumors seen in the MRTX849 single-dose group, it shows that the in vivo anti-tumor activity is enhanced by combining compound 1 with MRTX849.

Tumor proliferation inhibition rate (TGI%) = (1-TV/CV) × 100

Tumor volume (mm ³ ) = 1/2 × major diameter (mm) × minor diameter (mm) × minor diameter (mm)

Tumor volume change (mm ³ ) = tumor volume after day 0 - tumor volume on day 0

TV: the average value of tumor volume change in compound 1 group, MRTX849 group or combination group CV: the average value of tumor volume change in vehicle group

(Example 11 Evaluation of in vivo antitumor activity by combination of compound 1 and TNO155)

The in vivo antitumor activity of the combination of compound 1 and TNO155 was evaluated using a xenograft model transplanted with the lung cancer line NCI-H1373 (KRAS-G12C). 5×10 ⁶ cells were transplanted subcutaneously per nude mouse, and the nude mice were randomized when the tumor volume reached 200-300 mm ³ , and the administration of compound 1 and/or the combination compound was started (n=8). Compound 1 was orally administered once a day, and TNO155 was orally administered twice a day, and the administration period was 35 days. The dosage of the drug was 5 mg/kg for compound 1 and 5 mg/kg for TNO155. Compound 1 was dissolved in 10% DMSO/5% Cremphor EL/85% distilled water and administered, and TNO155 was dissolved in 0.5% Tween 80/0.5% Methylcellulose/99% distilled water and administered. Based on the tumor volume data on the final measurement day, the tumor growth inhibition rate (TGI%: tumor growth inhibition%) was calculated using the following formula. The results of the in vivo antitumor activity evaluation are shown in Figure 19. Compared with the vehicle group, the increase in tumor volume in the compound 1 single-dose administration group, the TNO155 single-dose administration group, and the compound 1 and TNO155 double-dose administration group was suppressed. Furthermore, in the comparison between the single-dose administration group and the double-dose administration group (combination group), since the tumor volume in the double-dose administration group was further reduced, it was shown that the in vivo antitumor activity was enhanced by the combination of compound 1 and TNO155.

Tumor proliferation inhibition rate (TGI%) = (1-TV/CV) × 100

Tumor volume (mm ³ ) = 1/2 × major diameter (mm) × minor diameter (mm) × minor diameter (mm)

Tumor volume change (mm ³ ) = tumor volume after day 0 - tumor volume on day 0

TV: the average value of tumor volume change in compound 1 group, TNO155 group or combination group CV: the average value of tumor volume change in vehicle group

(Example 12 Evaluation of in vivo antitumor activity by combination of Compound 1 and Compound 2)

The in vivo antitumor activity of the combination of compound 1 and compound 2 was evaluated using a xenograft model transplanted with lung cancer lines NCI-H441 (KRAS-G12C) or NCI-H1373 (KRAS-G12C). 5×10 ⁶ cells were transplanted subcutaneously per nude mouse, and randomization was performed when the tumor volume reached 200-300 mm ³ , and administration of compound 1 and/or the combination compound was started (n=5). Compound 1 and compound 2 were orally administered once a day for 14 days. The dosage of the drug was 3 mg/kg or 5 mg/kg for compound 1 and 0.25 mg/kg for compound 2. Compound 1 was dissolved in 10% DMSO/5% Cremphor EL/85% distilled water and administered, and compound 2 was dissolved in 10% DMSO/10% Cermphor EL/15% PEG400/15% HPBCD/50% distilled water and administered. According to the tumor volume data on the final measurement day, the tumor proliferation inhibition rate (TGI%: tumor growth inhibition%) was calculated using the following formula. The results of the in vivo antitumor activity evaluation are shown in Figures 20 and 21. Compared with the vehicle group, the increase in tumor volume in the compound 1 single-dose administration group, the compound 2 single-dose administration group, and the compound 1 and compound 2 double-dose administration group was inhibited. Furthermore, in the comparison between the single-dose administration group and the double-dose administration group (combination group), since the tumor volume in the double-dose administration group was further reduced, it was shown that the in vivo antitumor activity was enhanced by combining compound 1 with compound 2.

Tumor proliferation inhibition rate (TGI%) = (1-TV/CV) × 100

Tumor volume (mm ³ ) = 1/2 × major diameter (mm) × minor diameter (mm) × minor diameter (mm)

Tumor volume change (mm ³ ) = tumor volume after day 0 - tumor volume on day 0

TV: the average value of tumor volume change in compound 1 group, compound 2 group or combination group CV: the average value of tumor volume change in vehicle group

(Example 13 Evaluation of in vivo RAS signaling inhibitory activity by combination of compound 1 and compound 2)

After the in vivo antitumor activity evaluation in NCI-H441 of Example 12 was completed, Compound 1 and Compound 2 were further administered to nude mice (n=3) at 5 mg/kg and 0.25 mg/kg, respectively. Four hours later, the tumor was recovered and frozen in liquid nitrogen and stored at -80°C. Thereafter, the recovered xenograft samples were disrupted with Multi-Beads Shocker (Yasui Instruments) under liquid nitrogen cooling, and the RAS signal inhibitory activity of Compound 1 and Compound 2 in xenografts was evaluated based on the same protocol as in Example 1 (in vitro RAS signal inhibitory activity evaluation). The primary antibody, secondary antibody, dilution concentration, reaction time, and reaction temperature used are shown in Tables 10 and 11. The results of the electrophoresis diagram showing the results of the in vivo RAS signal inhibitory activity evaluation of Western blotting are shown in Figure 22. "KRAS-GTP" means "KRAS" bound to GTP, and "pERK", "pAKT" and "pMEK" respectively represent phosphorylated "ERK", "AKT" and "MEK". Compared with the case of single-dose application of Compound 1 or Compound 2, the case of combined use of Compound 1 and Compound 2 showed a decrease in pERK and pMEK, and enhanced inhibition of MAPK signaling.

[Table 10]

[Table 11]

Claims (26)

1. A drug for treating or preventing cancer, comprising a first active ingredient in combination with a second active ingredient. The first active ingredient is a compound represented by the following formula (1) or a salt thereof or a solvate thereof, and the second active ingredient is a molecular targeting agent. [Chemical formula 1] 2. A drug for treating or preventing cancer, comprising a first active ingredient in combination with a second active ingredient. The first active ingredient is a molecular targeting agent, and the second active ingredient is a compound represented by the following formula (1) or a salt thereof or a solvate thereof, [Chemical formula 2] 3. The drug according to claim 1 or 2, wherein the first active ingredient and the second active ingredient are provided as a kit. 4. The drug according to any one of claims 1 to 3, wherein the combination of the first active ingredient and the second active ingredient is administered simultaneously or separately. 5. The medicament according to any one of claims 1 to 4, wherein the first active ingredient and the second active ingredient are used in combination as a compound. 6. The drug according to any one of claims 1 to 5, wherein the molecular targeting agent is a MAPK/ERK pathway inhibitor. 7. The drug according to any one of claims 1 to 6, wherein the molecular targeting agent is at least one selected from the group consisting of an EGFR inhibitor, a VEGF inhibitor, a SHP2 inhibitor, a KRAS-G12C inhibitor, and a MEK inhibitor. 8. The drug according to any one of claims 1 to 7, wherein the molecular targeting agent is an EGFR inhibitor, a VEGF inhibitor, a SHP2 inhibitor, a KRAS-G12C inhibitor or a MEK inhibitor. 9. The drug according to claim 7 or 8, wherein the EGFR inhibitor is at least one selected from gefitinib, erlotinib, afatinib, osimertinib, lapatinib or salts thereof or solvates thereof, and anti-EGFR antibodies. 10. The drug according to any one of claims 7 to 9, wherein the EGFR inhibitor is an anti-EGFR antibody. 11. The drug according to claim 9 or 10, wherein the anti-EGFR antibody has a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 2. 12. The drug according to any one of claims 9 to 11, wherein the anti-EGFR antibody is cetuximab. 13. The drug according to any one of claims 7 to 12, wherein the VEGF inhibitor is an anti-VEGF antibody. 14 . The drug according to claim 13 , wherein the anti-VEGF antibody has a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 4. 15. The medicament according to claim 13 or 14, wherein the anti-VEGF antibody is bevacizumab. 16. The drug according to any one of claims 7 to 15, wherein the SHP2 inhibitor is at least one selected from the group consisting of RMC4550, TNO155, RLY1971, SHP099, NSC-87877, and salts thereof and solvates thereof. 17. The drug according to any one of claims 7 to 16, wherein the SHP2 inhibitor is at least one selected from the group consisting of RMC4550, TNO155, and salts thereof and solvates thereof. 18. The drug according to any one of claims 7 to 17, wherein the KRAS-G12C inhibitor is at least one selected from the group consisting of sotolacib, MRTX849, and salts thereof and solvates thereof. 19. The drug according to any one of claims 7 to 18, wherein the MEK inhibitor is at least one selected from the group consisting of the compound represented by the following formula (2), trametinib, selumetinib, CH4987655, and salts thereof and solvates thereof: [Chemical formula 3] 20. The drug according to any one of claims 7 to 19, wherein the MEK inhibitor is a compound represented by the following formula (2), trametinib, or a salt thereof, or a solvate thereof: [Chemical formula 4] 21. The drug according to any one of claims 1 to 20, wherein the compound represented by formula (1) or a salt thereof or a solvate thereof is a solvate of the compound represented by formula (1). 22. The medicament according to claim 21, wherein the solvate is a hydrate. 23. The drug according to any one of claims 1 to 20, wherein the compound represented by formula (1) or a salt thereof or a solvate thereof is the compound represented by formula (1). 24. The drug according to any one of claims 1 to 23, wherein the cancer is a solid cancer or a blood cancer. 25. The drug according to any one of claims 1 to 24, wherein the cancer is at least one selected from lung cancer, esophageal cancer, gastric cancer, colorectal cancer, uterine cancer, ovarian cancer, pancreatic cancer, bladder cancer, thyroid cancer, skin cancer, leukemia, malignant lymphoma and multiple myeloma. 26. The medicament according to any one of claims 1 to 25, wherein the cancer is a cancer associated with abnormality of the RAS gene or the MAPK/ERK pathway.