Biomarker for treatment of solid cancer by imidazo[4,5-b]pyridine derivative

Abstract

Provided is a means for predicting the anti-cancer therapeutic efficacy of 1-((5-(1R)-1-fluoroethyl)-1,3,4-oxadiazol-yl)methyl)-6-(4-methoxypyrrolo[2,1-f][1,2,4]triazin-5-yl)-2-methyl-1H-imidazo[4,5-b]pyridine or a derivative thereof in solid cancer, and for identifying a subject that is most likely to benefit from treatment using said compound and treatment using the compound in combination with other agents and / or other therapies.　As a biomarker for predicting a treatment response to the compound, the feature of a gene expected to impart sensitivity to CLK inhibition is used.

Description

Biomarkers for the treatment of solid cancers with imidazo[4,5-b]pyridine derivatives

[0001] The present invention relates to the use of the MYC gene and other cancer-associated genes as biomarkers for predicting therapeutic response to imidazo[4,5-b]pyridine derivatives in subjects with solid cancers, methods for diagnosing and treating solid cancers using the biomarkers, and pharmaceutical compositions for treating solid cancers characterized by the biomarkers.

[0002] Imidazo[4,5-b]pyridine derivatives, in particular compounds of the formula (I) 1-((5-(1R)-1-fluoroethyl)-1,3,4-oxadiazol-yl)methyl)-6-(4-methoxypyrrolo[2,1-f][1,2,4]triazin-5-yl)-2-methyl-1H-imidazo[4,5-b]pyridine (hereinafter referred to as Compound A), represented by the formula: or a derivative thereof, is an inhibitor of CLK (Cdc2-like kinase) that induces mainly skipped-exon splicing changes and exhibits potent antitumor activity. Patent Document 1 discloses that CLK inhibitors including Compound A or its derivatives are useful for the prevention and / or treatment of CLK-related diseases, including cancer, and that they exhibit antitumor effects in various in vivo cancer models, including ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer, blood cancer, and prostate cancer.

[0003] International Publication No. 2017 / 188374

[0004] The present invention aims to predict the anti-cancer therapeutic efficacy of Compound A or a derivative thereof in solid tumors, thereby identifying subjects who would most benefit from treatment with Compound A or a derivative thereof and treatment using Compound A or a derivative thereof in combination with other drugs and / or other therapies.

[0005] The present inventors have discovered a gene characteristic that is predicted to confer sensitivity to CLK inhibition, which shows particularly high responsiveness to Compound A or its derivatives, in solid tumors. Based on this finding, they have conducted extensive research and completed the present invention. Specifically, the present invention is as follows: [1] A method for the treatment of a solid tumor subject with a compound of the following formula (I):1. A method for predicting a therapeutic response to 1-((5-(1R)-1-fluoroethyl)-1,3,4-oxadiazol-yl)methyl)-6-(4-methoxypyrrolo[2,1-f][1,2,4]triazin-5-yl)-2-methyl-1H-imidazo[4,5-b]pyridine of Formula (I) or a derivative thereof, comprising: (1) detecting a genetic signature predicted to confer sensitivity to sirtuin-activating factor (CLK) inhibition in a sample derived from cancer tissue from the subject; and (2) determining the likelihood of the subject responding to the compound of Formula (I) or a derivative thereof based on the genetic signature. [2] The method according to the above item [1], wherein the gene characteristic is one or more selected from the group consisting of: (a) MYC gene amplification, (b) MYC gene mutation, (c) MYC gene chromosomal rearrangement, (d) increased MYC mRNA amount, (e) increased MYC protein amount, (f) EGFR gene mutation, (g) U2AF1 gene mutation, (h) POLE gene loss, (i) CDK12 gene loss, (j) gene mutation selected from one or more of ATRX, ATR, GNAQ, and HIST1H3B, and (k) MDM4 exon skipping change. [3] The method according to item [2], comprising: (1) detecting MYC gene amplification, MYC gene mutation, MYC gene chromosomal location rearrangement, increased MYC mRNA level, and / or increased MYC protein level in a sample derived from cancer tissue from the subject, and (2) determining that the subject is particularly responsive to the compound represented by formula (I) or a derivative thereof when the MYC gene copy number is increased compared to a reference MYC gene copy number, the MYC gene sequence is altered compared to a reference MYC gene sequence, the MYC gene chromosomal location site is translocated compared to a reference MYC gene chromosomal location site, the MYC mRNA level is increased compared to the reference MYC mRNA level, and / or the MYC protein level is increased compared to the reference MYC protein level. [4] The method according to item [3], wherein the MYC gene is selected from the group consisting of MYCC, MYCN, and MYCL.[5] The method according to item [3], wherein the MYC gene copy number is increased by about 1.5-fold or more compared to the reference MYC gene copy number, or the MYC gene sequence is MYCC Q37del, P59L, S146L, or MYCN R45Rfs*86, E445K compared to the reference. [6] The method according to item [3], wherein the MYC gene amplification or MYC gene mutation is detected by array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing, next-generation sequencing (NGS), or multiplex ligation-dependent probe amplification (MLPA). [7] The method according to the above item [3], comprising detecting the rearrangement of the MYC gene chromosomal location site by in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next-generation sequencing (NGS), or comparative genomic hybridization (CGH). [8] The method according to the above item [3], comprising detecting the increase in the amount of MYC mRNA by in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next-generation sequencing (NGS), or real-time PCR or digital PCR analysis. [9] The method according to item [3], comprising detecting the increase in the amount of MYC protein by immunohistochemical staining (IHC), flow cytometry (FACS), enzyme-linked immunosorbent assay (ELISA), proteinarray (Proximity Extension Assay, PEA method), or Western blotting (WB).

[10] The method according to item [2], comprising: (1) detecting a gene mutation selected from one or more of EGFR gene mutation, U2AF1 gene mutation, POLE gene reduction, CDK12 gene reduction, ATRX, ATR, GNAQ, and HIST1H3B, and / or MDM4 exon skipping alteration in a sample derived from cancer tissue from the subject; and (2) determining that the subject is particularly responsive to the compound of formula (I) or a derivative thereof when a gene sequence selected from one or more of EGFR gene, U2AF1 gene, ATRX, ATR, GNAQ, and HIST1H3B is altered compared to a reference gene sequence of each gene, and / or when an MDM4 mRNA sequence is altered compared to a reference MDM4 mRNA sequence, and / or when the POLE gene copy number or CDK12 gene copy number is reduced compared to a reference copy number of each gene.

[11] The method according to item

[10] , wherein the U2AF1 gene mutation is an S34F mutation or an S34Y mutation.

[12] The method according to item

[10] or

[11] , wherein the EGFR gene mutation, the U2AF1 gene mutation, the POLE gene loss, the CDK12 gene loss, the gene mutation selected from one or more of ATRX, ATR, GNAQ, and HIST1H3B, and / or the MDM4 exon skipping alteration is detected by array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing or multiplex ligation-dependent probe amplification (MLPA), or RNA sequencing.

[13] The method according to any one of the above items [1] to

[12] , wherein the cancer tissue-derived sample is a surgical sample, biopsy sample, pleural effusion sample, or ascites sample obtained from the subject, for example, collected from one or more of lung, blood, breast, digestive tract (stomach, colon, rectum), ovary, pancreas, liver, bladder, cervix, nerve, uterus, salivary gland, kidney, prostate, thyroid, or muscle tissue.

[14] The method according to any one of the above items [1] to

[12] , wherein the subject is a human subject.

[15] The method according to any one of the above items [1] to

[12] , wherein the cancer is selected from the group consisting of carcinoma, rhabdomyosarcoma, sarcoma such as alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fiber, blood vessel, adipose, and / or connective tissue), clear cell sarcoma, neuroblastoma, medulloblastoma, astrocytoma, glioblastoma multiforme, retinoblastoma, adenosquamous carcinoma, carcinosarcoma,

[16] The method according to any one of items [3] to [9], wherein the MYC gene is MYCC and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, gastrointestinal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

[17] The method according to item

[16] , wherein the MYC gene is MYCC and the cancer is ovarian cancer.

[18] The method according to any one of items [3] to [9], wherein the MYC gene is MYCN, and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, prostate cancer, neuroblastoma, small cell lung cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, and retinoblastoma.

[19] The method according to item

[18] , wherein the MYC gene is MYCN, and the cancer is neuroblastoma.

[20] The method according to any one of items [3] to [9], wherein the MYC gene is MYCL, and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, and prostate cancer.

[21] In a subject with a solid cancer, a method comprising administering to a subject a compound of formula (I): a biomarker selected from one or more of the following (a) to (k) for predicting a therapeutic response to a therapy with 1-((5-(1R)-1-fluoroethyl)-1,3,4-oxadiazol-yl)methyl)-6-(4-methoxypyrrolo[2,1-f][1,2,4]triazin-5-yl)-2-methyl-1H-imidazo[4,5-b]pyridine or a derivative thereof, represented by the formula: (k) MDM4 exon skipping alteration.

[22] The biomarker according to item

[21] , wherein the MYC gene is selected from the group consisting of MYCC, MYCN, and MYCL.

[23] The biomarker according to item

[21] , wherein the mutation in the U2AF1 gene is an S34F mutation or an S34Y mutation.

[24] The biomarker according to any one of items

[21] to

[23] , wherein the cancer is a carcinoma, a rhabdomyosarcoma, a sarcoma such as alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fiber, blood vessels, adipose, and / or connective tissue), clear cell sarcoma, neuroblastoma, medulloblastoma, astrocytoma, glioblastoma multiforme, retinoblastoma, adenosquamous carcinoma, or carcinosarcoma.

[25] The biomarker according to item

[21] , wherein the MYC gene is MYCC, and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney cancer or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

[26] The biomarker according to item

[25] , wherein the MYC gene is MYCC and the cancer is ovarian cancer.

[27] The biomarker according to item

[21] , wherein the MYC gene is MYCN and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, prostate cancer, neuroblastoma, small cell lung cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, and retinoblastoma.

[28] The biomarker according to item

[27] , wherein the MYC gene is MYCN and the cancer is neuroblastoma.

[29] The biomarker according to item

[21] , wherein the MYC gene is MYCL and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, and prostate cancer.

[30] A compound of formula (I): 1. A pharmaceutical composition comprising 1-((5-(1R)-1-fluoroethyl)-1,3,4-oxadiazol-yl)methyl)-6-(4-methoxypyrrolo[2,1-f][1,2,4]triazin-5-yl)-2-methyl-1H-imidazo[4,5-b]pyridine represented by the formula: or a derivative thereof as an active ingredient, wherein the gene characteristic is one or more selected from the following (a) to (k): (a) MYC gene amplification, (b) MYC gene mutation, (c) MYC gene chromosomal location rearrangement, (d) increased MYC mRNA amount, (e) increased MYC protein amount, (f) EGFR gene mutation, (g) U2AF1 gene mutation, (h) POLE gene reduction, (i) CDK12 gene reduction, (j) a gene mutation selected from one or more of ATRX, ATR, GNAQ, and HIST1H3B, and (k) an MDM4 exon skipping change.

[31] The pharmaceutical composition according to item

[30] , wherein the MYC gene is selected from the group consisting of MYCC, MYCN, and MYCL.

[32] The pharmaceutical composition according to item

[30] , wherein the U2AF1 gene mutation is an S34F mutation or an S34Y mutation.

[33] The pharmaceutical composition according to any one of the above items

[30] to

[32] , wherein the cancer is a carcinoma, a rhabdomyosarcoma, a sarcoma such as, for example, alveolar rhabdomyosarcoma (including a sarcoma derived from bone, tendon, cartilage, muscle, fat, fiber, blood vessel, adipose, and / or connective tissue), a clear cell sarcoma, a neuroblastoma, a medulloblastoma, an astrocytoma, a glioblastoma multiforme, a retinoblastoma, an adenosquamous carcinoma, or a carcinosarcoma.

[34] The pharmaceutical composition according to item

[30] , wherein the MYC gene is MYCC, and the cancer is selected from one or more of ovarian cancer, breast cancer, mixed mesodermal tumor, teratocarcinoma, lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung cancer), breast cancer (including metastatic breast cancer), gastrointestinal cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney cancer or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

[35] The pharmaceutical composition according to item

[34] , wherein the MYC gene is MYCC and the cancer is ovarian cancer.

[36] The pharmaceutical composition according to item

[30] , wherein the MYC gene is MYCN and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, neuroblastoma, small cell lung cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, and retinoblastoma.

[37] The pharmaceutical composition according to item

[36] , wherein the MYC gene is MYCN and the cancer is neuroblastoma.

[38] The pharmaceutical composition according to item

[30] , wherein the MYC gene is MYCL and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, and prostate cancer.

[0006] The present invention provides a biomarker for predicting the therapeutic efficacy of Compound A or its derivatives in subjects with solid cancer, thereby making it possible to identify subjects who may achieve a higher anti-cancer effect in therapy with Compound A or its derivatives.

[0007]

[0023] Figure 1 shows an evaluation of the sensitivity of Compound A to NIH3T3 cell lines introduced with various cancer-related genes. Figure 2 shows an evaluation of the sensitivity of Compound A to NIH3T3 cell lines having genetic characteristics in MYCC or MYCN. Figure 3 shows an evaluation of the sensitivity of Compound A to cell lines having genetic characteristics in MYCC, MYCN, or EGFR. Figure 4 shows an evaluation of the sensitivity of Compound A to cell lines having MYCC gene amplification. Figures 1a) and 1b show powder X-ray diffraction, differential scanning calorimetry, and thermogravimetry, showing an example of a crystal of Compound A or a derivative thereof (crystal pattern A). Figures 1a) and 1b show powder X-ray diffraction, differential scanning calorimetry, and thermogravimetry, showing an example of a crystal of Compound A or a derivative thereof (crystal pattern B).

[0008] Hereinafter, the aspects of the present invention will be described in detail, including a method for predicting the therapeutic response of solid cancer to compound A or a derivative thereof, a biomarker thereof, a pharmaceutical composition containing compound A or a derivative thereof for treating solid cancer characterized by the biomarker, and a method for diagnosing and treating the solid cancer.

[0009] [Compound A] Within the scope of the present invention, Compound A is a compound of the following formula (I): This represents 1-((5-(1R)-1-fluoroethyl)-1,3,4-oxadiazol-yl)methyl)-6-(4-methoxypyrrolo[2,1-f][1,2,4]triazin-5-yl)-2-methyl-1H-imidazo[4,5-b]pyridine represented by the formula:

[0010] Furthermore, within the scope of the present invention, derivatives of Compound A include salts and hydrates of the compound of formula (I), as well as optical isomers, tautomers, solvates, etc. of these. Furthermore, this derivative of Compound A may be a crystal or clathrate, and may have a single crystalline form or may be a crystalline mixture, co-crystal, etc. As the salt of Compound A, pharmacologically acceptable salts are preferred, and examples thereof include salts with inorganic bases, salts with organic bases, salts with inorganic acids, salts with organic acids, and salts with basic or acidic amino acids. Suitable examples of salts with inorganic bases include alkali metal salts such as sodium salts and potassium salts; alkaline earth metal salts such as calcium salts and magnesium salts; aluminum salts, and ammonium salts. Suitable examples of salts with organic bases include salts with trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine, tromethamine [tris(hydroxymethyl)methylamine], tert-butylamine, cyclohexylamine, benzylamine, dicyclohexylamine, and N,N-dibenzylethylenediamine. Suitable examples of salts with inorganic acids include salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, and phosphoric acid. Suitable examples of salts with organic acids include salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid. Suitable examples of salts with basic amino acids include salts with arginine, lysine, and ornithine. Suitable examples of salts with acidic amino acids include salts with aspartic acid and glutamic acid.

[0011] In addition, details of the method for producing Compound A or a derivative thereof are disclosed in International Publication No. 2017 / 188374, the contents of which are incorporated herein by reference.

[0012] [Method for predicting therapeutic response to Compound A in solid tumors] The method of the present invention for predicting therapeutic response to Compound A or a derivative thereof in solid tumors comprises: (a) detecting a genetic signature predicted to confer sensitivity to sirtuin-activating factor (CLK) inhibition in a cancer tissue-derived sample from a subject with a solid tumor; and (b) determining the likelihood of the subject responding to Compound A or a derivative thereof based on the genetic signature.

[0013] The genetic features predicted to confer sensitivity to sirtuin inhibitors include genetic alterations and gene expression changes, where the genetic alterations may include mutations in the genomic region of a gene, such as copy number variations (CNVs), single nucleotide variations (SNVs), multiple nucleotide variations (MNVs), small insertions or deletions (Indels), splice site mutations, gene fusions or chromosomal site rearrangements, or RNA splicing alterations. Examples of the genes include ACVR1, AKT1, AKT2, AKT3, ALK, AR, ARAF, BMPR1A, BRAF, CASP8, CCNE1, CDK4, CSF1R, CTNNB1, DDR2, EPAS1, ERBB2, ERBB3, ERBB4, ESR1, EZH2, GNA11, GNAQ, GNAS, H3F3A, HRAS, IDH1, IDH2, JUN, KIT, KLF4, KRAS, MAP2K2, MAP2K4, MAPK1, MDM2, MDM4, and MET. , MITF, MYCC, MYCN, MYCL, MYOD1, MFE2L2, MKX2-1, NRAS, NTRK1, PDGFRA, PIK3CA, PIK3CB, PLCG1, PTEN, RAC1, RAF1, RET, RHOA, SMO, STAT5B, TBL1XR1, TERT, TP53, TSC2, USP8, WT1, EGFR, FGFR1, FGFR2, FGFR3, MAP2K1, U2AF1, ATR, ATRX, and HIST1H3B. The gene expression change includes an increase or decrease in mRNA amount or protein amount, which may or may not be based on the gene change.

[0014] In one embodiment of the present invention, the genetic feature predicted to confer sensitivity to sirtuin-activating factor (CLK) inhibition may be MYC gene amplification, MYC gene mutation, or MYC gene chromosomal location rearrangement, each of which is detected by comparison with a respective reference. Preferably, a subject can be determined to be particularly responsive to Compound A or a derivative thereof if the MYC gene copy number is increased compared to the MYC gene copy number reference, the MYC gene sequence is altered compared to the MYC gene sequence reference, or the MYC gene chromosomal location is rearranged compared to the MYC gene chromosomal location reference.

[0015] The MYC gene amplification may be any amplification that results in an observed increase in copy number compared to a reference, and may preferably be, but is not limited to, an increase of about 1.5-fold or more. In other embodiments of the present invention, the increase may be, but is not limited to, an increase of about 2-fold or more, about 4-fold or more, about 10-fold or more, or about 20-fold or more.

[0016] The MYC gene mutation may be a mutation such as a single nucleotide variant (SNV), a multiple nucleotide variant (MNV), a small insertion or deletion (Indel), a splice site mutation, or a gene fusion within the genomic region of the MYC gene.

[0017] The MYC gene mutation may be, but is not limited to, preferably MYCC Q37del, P59L, S146L, MYCN R45Rfs*86 and / or E445K.

[0018] In a preferred embodiment of the present invention, the MYC gene may be selected from MYCC, MYCN and MYCL.

[0019] In one embodiment of the present invention, the MYC gene amplification or MYC gene mutation can be detected by a detection method well known in the art, such as array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing, copy number variation (CNV) analysis by real-time PCR or digital PCR, next-generation sequencing (NGS), or multiplex ligation-dependent probe amplification (MLPA).

[0020] Similarly, rearrangements of the MYC gene chromosomal location site can be detected by detection techniques well known in the art, such as in situ hybridization (ISH), fluorescence in situ hybridization (FISH), next-generation sequencing (NGS), or comparative genomic hybridization (CGH). The MYC gene copy number reference, MYC gene sequence reference, and / or MYC gene chromosomal location site reference may be obtained from a sample derived from a non-cancerous tissue of the same subject, or may be obtained from a database well known in the art, such as cBioPortal FOR CANCER GENOMICS.

[0021] In one embodiment of the present invention, the genetic feature predicted to confer sensitivity to sirtuin-activating factor (CLK) inhibition may be an increased MYC mRNA level or an increased MYC protein level, which is detected by comparison with a reference. Preferably, if the MYC mRNA level is increased compared to the reference MYC mRNA level or the MYC protein level is increased compared to the reference MYC protein level, the subject can be determined to be particularly responsive to Compound A or a derivative thereof.

[0022] The increase in the amount of MYC mRNA and / or the increase in the amount of MYC protein may be due to, but is not limited to, MYC gene amplification, MYC gene mutation, and / or MYC gene chromosomal location site rearrangement.

[0023] The increase in the amount of MYC mRNA may be observed as an increase in the amount of mRNA compared to a reference, and the increase in the amount of MYC protein may be observed as an increase in the amount of protein compared to a reference.

[0024] The amount of MYC mRNA can be detected by detection techniques well known in the art, such as in situ hybridization (ISH), fluorescence in situ hybridization (FISH), next-generation sequencing (NGS), or real-time PCR or digital PCR analysis.

[0025] The reference MYC mRNA level may be obtained from a sample derived from a non-cancerous tissue from the same subject, or may be obtained from a database well known in the art, such as cBioPortal FOR CANCER GENOMICS.

[0026] The amount of the MYC protein can be detected by detection techniques well known in the art, such as immunohistochemical staining (IHC), flow cytometry (FACS), enzyme-linked immunosorbent assay (ELISA), proteinarray (Proximity Extension Assay, PEA method), and Western blotting (WB).

[0027] The reference MYC protein amount may be obtained from a sample derived from a non-cancerous tissue from the same subject, or may be obtained from a database well known in the art, such as cBioPortal FOR CANCER GENOMICS.

[0028] In one embodiment of the present invention, the genetic feature predicted to confer sensitivity to sirtuin inhibitors is a U2AF1 gene mutation, which is detected by comparison with a reference. Preferably, if the genetic sequence of the U2AF1 gene is altered compared to the reference U2AF1 gene sequence, the subject can be determined to be particularly responsive to Compound A or a derivative thereof.

[0029] The U2AF1 gene mutation may be a mutation such as a single nucleotide variation (SNV), a multiple nucleotide variation (MNV), a small insertion or deletion (Indel), a splice site mutation, or a gene fusion within the genomic region of the gene, but is preferably an S34F mutation or an S34Y mutation.

[0030] In one embodiment of the present invention, the genetic feature predicted to confer sensitivity to sirtuin-activating factor (CLK) inhibition is an EGFR gene mutation, a POLE gene loss, a CDK12 gene loss, a gene mutation selected from one or more of ATRX, ATR, GNAQ, and HIST1H3B, and / or an MDM4 exon skipping change, each of which is detected by comparison with a respective reference. Preferably, the subject can be determined to be particularly responsive to Compound A or a derivative thereof if the gene sequence of the EGFR gene is altered compared to the EGFR gene sequence reference, the POLE gene copy number is decreased compared to the POLE gene copy number reference, the CDK12 gene copy number is decreased compared to the CDK12 gene copy number reference, the gene sequence of one or more of ATRX, ATR, GNAQ, and HIST1H3B is altered compared to the corresponding gene sequence reference, and / or the MDM4 mRNA sequence is altered compared to the MDM4 mRNA sequence reference.

[0031] The gene mutation selected from one or more of EGFR gene mutation, ATRX, ATR, GNAQ, and HIST1H3B and / or MDM4 exon skipping variation may each be a mutation such as a single nucleotide variation (SNV), a multiple nucleotide variation (MNV), a small insertion or deletion (Indel), a splice site mutation, or a gene fusion, and / or an RNA splicing variation.

[0032] The POLE gene reduction or CDK12 gene reduction may be one in which a reduction in copy number is observed compared to the respective references.

[0033] The cancer tissue-derived sample may be a surgical sample, biopsy sample, pleural effusion sample, or ascites sample obtained from the subject, and may be taken from one or more of lung, blood, breast, gastrointestinal tract (stomach, colon, rectum), ovary, pancreas, liver, bladder, cervix, nerve, uterus, salivary gland, kidney, prostate, thyroid, or muscle tissue.

[0034] The subject is preferably, but not limited to, a human subject.

[0035] The solid tumors include cancers [e.g., colorectal cancer (e.g., colon cancer, rectal cancer, anal cancer, familial colorectal cancer, hereditary non-polyposis colorectal cancer, gastrointestinal stromal tumor), lung cancer (e.g., non-small cell lung cancer, small cell lung cancer, malignant mesothelioma), mesothelioma, pancreatic cancer (e.g., pancreatic ductal carcinoma, pancreatic endocrine tumor), pharyngeal cancer, laryngeal cancer, esophageal cancer, gastric cancer (e.g., papillary adenocarcinoma, mucinous adenocarcinoma, adenosquamous carcinoma), duodenal cancer, small intestinal cancer, breast cancer (e.g., invasive ductal carcinoma, ductal carcinoma in situ, inflammatory breast cancer), ovarian cancer (e.g., epithelial ovarian cancer, extragonadal germ cell tumor, ovarian germ cell tumor, ovarian low malignant potential tumor), testicular tumor, prostate cancer (e.g., hormone-dependent prostate cancer, hormone-independent prostate cancer, castration therapy-resistant prostate cancer), ), liver cancer (e.g., hepatocellular carcinoma, primary liver cancer, extrahepatic bile duct cancer), thyroid cancer (e.g., medullary thyroid carcinoma), kidney cancer (e.g., renal cell carcinoma (e.g., clear cell renal cell carcinoma), transitional cell carcinoma of the renal pelvis and ureter), uterine cancer (e.g., cervical cancer, uterine cancer, uterine sarcoma), gestational choriocarcinoma, brain tumor (e.g., medulloblastoma, glioma, pineal astrocytoma, pilocytic astrocytoma, diffuse astrocytoma, anaplastic astrocytoma, pituitary adenoma), retinoblastoma, skin cancer (e.g., basal cell carcinoma, malignant melanoma), sarcoma (e.g., rhabdomyosarcoma, leiomyosarcoma, soft tissue sarcoma, spindle cell sarcoma, clear cell sarcoma), malignant bone tumor, bladder cancer, cancer of unknown primary origin).

[0036] In a preferred embodiment of the present invention, the cancer may be selected from one or more of carcinoma, rhabdomyosarcoma, sarcoma such as, for example, alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fiber, blood vessels, adipose, and / or connective tissue), clear cell sarcoma, neuroblastoma, medulloblastoma, astrocytoma, glioblastoma multiforme, retinoblastoma, adenosquamous carcinoma, carcinosarcoma, mixed mesodermal tumor, teratocarcinoma, lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung carcinoma), breast cancer (including metastatic breast cancer), gastrointestinal cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney cancer or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

[0037] In a preferred embodiment of the present invention, the genetic characteristic may be MYCC gene amplification, MYCC gene mutation, or MYCC gene chromosomal location rearrangement in a sample derived from cancer tissue of a solid cancer selected from ovarian cancer, gastric cancer, lung cancer, breast cancer, hepatocellular carcinoma, head and neck squamous cell carcinoma, rectal cancer, bile duct cancer, pancreatic cancer, uveal melanoma, sarcoma, prostate cancer, colon cancer, cutaneous melanoma, mesothelioma, glioblastoma, glioma, cervical squamous cell carcinoma and cervical cancer, testicular germ cell tumor, thymoma, thyroid cancer, renal cell carcinoma, renal papillary cell carcinoma, adrenocortical carcinoma, pheochromocytoma, esophageal cancer, uterine carcinosarcoma, lung squamous cell carcinoma, and bladder urothelial carcinoma (Cell Syst. 2018 Mar 28; 6(3): 282-300.e2).

[0038] In a particularly preferred embodiment of the present invention, the genetic characteristic may be MYCC gene amplification, MYCC gene mutation, or MYCC gene chromosomal location rearrangement in a sample derived from cancer tissue of ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer, or prostate cancer.

[0039] In a preferred embodiment of the present invention, the genetic characteristic may be MYCN gene amplification, MYCN gene mutation, or MYCN gene chromosomal location rearrangement in a sample derived from cancer tissue of a solid cancer selected from gastric cancer, lung cancer, breast cancer, hepatocellular carcinoma, head and neck squamous cell carcinoma, cholangiocarcinoma, uveal melanoma, sarcoma, prostate cancer, colon cancer, cutaneous melanoma, mesothelioma, glioblastoma, glioma, cervical squamous cell carcinoma and cervical cancer, testicular germ cell tumor, adrenocortical carcinoma, pheochromocytoma, esophageal cancer, uterine carcinosarcoma, lung squamous cell carcinoma, and bladder urothelial carcinoma (Cell Syst. 2018 Mar 28; 6(3): 282-300.e2).

[0040] In a preferred embodiment of the present invention, the genetic characteristic may be MYCN gene amplification, MYCN gene mutation, or MYCN gene chromosomal location rearrangement in a sample derived from cancer tissue of a solid cancer selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, neuroblastoma, small cell lung cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, and retinoblastoma.

[0041] In a particularly preferred embodiment of the present invention, the genetic characteristic may be MYCN gene amplification, MYCN gene mutation, or MYCN gene chromosomal location rearrangement in a sample derived from neuroblastoma tissue.

[0042] In a preferred embodiment of the present invention, the genetic characteristic may be MYCL gene amplification, MYCL gene mutation, or MYCL gene chromosomal site rearrangement in a sample derived from cancer tissue of a solid cancer selected from ovarian cancer, gastric cancer, lung cancer, breast cancer, hepatocellular carcinoma, head and neck squamous cell carcinoma, rectal cancer, bile duct cancer, uveal melanoma, sarcoma, prostate cancer, colon cancer, cutaneous melanoma, mesothelioma, glioblastoma, glioma, cervical squamous cell carcinoma and cervical cancer, testicular germ cell tumor, thymoma, adrenocortical carcinoma, esophageal cancer, uterine carcinosarcoma, lung squamous cell carcinoma, and bladder urothelial carcinoma (Cell Syst. 2018 Mar 28; 6(3): 282-300.e2).

[0043] In a preferred embodiment of the present invention, the genetic characteristic may be MYCL gene amplification, MYCL gene mutation, or MYCL gene chromosomal location rearrangement in a sample derived from cancer tissue of a solid cancer selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, and prostate cancer.

[0044] A further embodiment of the present invention may include administering a therapy using Compound A or a derivative thereof to a subject in need of treatment for a solid tumor according to the predicted result of the treatment response. The therapy may include administering Compound A or a derivative thereof to the subject. In addition to administering Compound A or a derivative thereof alone, the therapy may also include administering Compound A or a derivative thereof in combination with other drugs and / or in combination with other therapies.

[0045] [Biomarker] The biomarker of the present invention is one or more biomarkers selected from the following (a) to (k) for predicting a therapeutic response to a therapy using Compound A or a derivative thereof in a subject with a solid cancer; (a) MYC gene amplification, (b) MYC gene mutation, (c) MYC gene chromosomal location rearrangement, (d) increased MYC mRNA amount, (e) increased MYC protein amount, (f) EGFR gene mutation, (g) U2AF1 gene mutation, (h) POLE gene reduction, (i) CDK12 gene reduction, (j) gene mutation selected from one or more of ATRX, ATR, GNAQ and HIST1H3B, and (k) MDM4 exon skipping alteration.

[0046] The MYC gene amplification, MYC gene mutation, or MYC gene chromosomal location site rearrangement is detected by comparison with each reference. Preferably, the subject can be determined to be particularly responsive to compound A or a derivative thereof when the MYC gene copy number is increased compared to the MYC gene copy number reference, the gene sequence of the MYC gene is changed compared to the MYC gene sequence reference, or the MYC gene chromosomal location site is rearranged compared to the MYC gene chromosomal location site reference.

[0047] The MYC gene amplification may be observed to result in an increase in copy number compared to a reference, and may preferably, but not limited to, show an increase of about 1.5-fold or more. In other embodiments of the present invention, the increase may be, but is not limited to, an increase of about 2-fold or more, about 4-fold or more, about 10-fold or more, or about 20-fold or more.

[0048] The MYC gene mutation may be a mutation such as a single nucleotide variant (SNV), a multiple nucleotide variant (MNV), a small insertion or deletion (Indel), a splice site mutation, or a gene fusion within the genomic region of the MYC gene.

[0049] The MYC gene mutation may be, but is not limited to, preferably MYCC Q37del, P59L, S146L, MYCN R45Rfs*86 and / or E445K.

[0050] In a preferred embodiment of the present invention, the MYC gene may be selected from the group consisting of MYCC, MYCN, and MYCL.

[0051] The increase in the MYC mRNA amount or the increase in the MYC protein amount is detected by comparison with a reference. Preferably, when the MYC mRNA amount is increased compared to the reference MYC mRNA amount or the MYC protein amount is increased compared to the reference MYC protein amount, the subject can be determined to be a subject that shows particularly high responsiveness to compound A or a derivative thereof.

[0052] The increase in the amount of MYC mRNA or the increase in the amount of MYC protein may be due to, but is not limited to, MYC gene amplification, MYC gene mutation, or MYC gene chromosomal site rearrangement.

[0053] The U2AF1 gene mutation is detected by comparison with a reference. Preferably, if the gene sequence of the U2AF1 gene is altered compared with the reference U2AF1 gene sequence, the subject can be determined to be a subject that shows particularly high responsiveness to compound A or a derivative thereof.

[0054] The U2AF1 gene mutation may be a mutation such as a single nucleotide variation (SNV), a multiple nucleotide variation (MNV), a small insertion or deletion (Indel), a splice site mutation, or a gene fusion within the genomic region of the gene, but is preferably an S34F mutation or an S34Y mutation.

[0055] The EGFR gene mutation, POLE gene loss, CDK12 gene loss, gene mutation selected from one or more of ATRX, ATR, GNAQ, and HIST1H3B, and / or MDM4 exon skipping change are detected by comparison with the respective references. Preferably, the subject can be determined to be particularly responsive to compound A or a derivative thereof when the gene sequence of the EGFR gene is altered compared to the EGFR gene sequence reference, the POLE gene copy number is decreased compared to the POLE gene copy number reference, the CDK12 gene copy number is decreased compared to the CDK12 gene copy number reference, the gene sequence of a gene selected from one or more of ATRX, ATR, GNAQ, and HIST1H3B is altered compared to the corresponding gene sequence reference, and / or the MDM4 mRNA sequence is altered compared to the MDM4 mRNA sequence reference.

[0056] The gene mutation selected from one or more of EGFR gene mutation, ATRX, ATR, GNAQ, and HIST1H3B, and / or MDM4 exon skipping variation may each be a mutation such as a single nucleotide variation (SNV), a multiple nucleotide variation (MNV), a small insertion or deletion (Indel), a splice variation, or a gene fusion, and / or an RNA splicing variation.

[0057] The POLE gene reduction or CDK12 gene reduction may be one in which a reduction in copy number is observed in comparison with the respective references.

[0058] The references used for detecting the biomarkers may be obtained from non-cancerous tissue samples from the same subject or from databases well known in the art. The detection method may be any of those well known in the art, and is not limited thereto, but preferably, the detection methods described above can be used.

[0059] The subject is preferably, but not limited to, a human subject.

[0060] The solid tumors include cancers [e.g., colorectal cancer (e.g., colon cancer, rectal cancer, anal cancer, familial colorectal cancer, hereditary non-polyposis colorectal cancer, gastrointestinal stromal tumor), lung cancer (e.g., non-small cell lung cancer, small cell lung cancer, malignant mesothelioma), mesothelioma, pancreatic cancer (e.g., pancreatic ductal carcinoma, pancreatic endocrine tumor), pharyngeal cancer, laryngeal cancer, esophageal cancer, gastric cancer (e.g., papillary adenocarcinoma, mucinous adenocarcinoma, adenosquamous carcinoma), duodenal cancer, small intestinal cancer, breast cancer (e.g., invasive ductal carcinoma, ductal carcinoma in situ, inflammatory breast cancer), ovarian cancer (e.g., epithelial ovarian cancer, extragonadal germ cell tumor, ovarian germ cell tumor, ovarian low malignant potential tumor), testicular tumor, prostate cancer (e.g., hormone-dependent prostate cancer, hormone-independent prostate cancer, castration therapy-resistant prostate cancer), ), liver cancer (e.g., hepatocellular carcinoma, primary liver cancer, extrahepatic bile duct cancer), thyroid cancer (e.g., medullary thyroid carcinoma), kidney cancer (e.g., renal cell carcinoma (e.g., clear cell renal cell carcinoma), transitional cell carcinoma of the renal pelvis and ureter), uterine cancer (e.g., cervical cancer, uterine cancer, uterine sarcoma), gestational choriocarcinoma, brain tumor (e.g., medulloblastoma, glioma, pineal astrocytoma, pilocytic astrocytoma, diffuse astrocytoma, anaplastic astrocytoma, pituitary adenoma), retinoblastoma, skin cancer (e.g., basal cell carcinoma, malignant melanoma), sarcoma (e.g., rhabdomyosarcoma, leiomyosarcoma, soft tissue sarcoma, spindle cell sarcoma, clear cell sarcoma), malignant bone tumor, bladder cancer, cancer of unknown primary origin).

[0061] In a preferred embodiment of the present invention, the solid cancer may be selected from one or more of carcinoma, rhabdomyosarcoma, sarcoma such as, for example, alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fiber, blood vessels, adipose, and / or connective tissue), clear cell sarcoma, neuroblastoma, medulloblastoma, astrocytoma, glioblastoma multiforme, retinoblastoma, adenosquamous carcinoma, carcinosarcoma, mixed mesodermal tumor, teratocarcinoma, lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung carcinoma), breast cancer (including metastatic breast cancer), gastrointestinal cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney cancer or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

[0062] In a preferred embodiment of the present invention, the biomarkers may be MYCC gene amplification, MYCC gene mutation, or MYCC gene chromosomal location rearrangement in solid cancers selected from ovarian cancer, gastric cancer, lung cancer, breast cancer, hepatocellular carcinoma, head and neck squamous cell carcinoma, rectal cancer, bile duct carcinoma, pancreatic cancer, uveal melanoma, sarcoma, prostate cancer, colon cancer, cutaneous melanoma, mesothelioma, glioblastoma, glioma, cervical squamous cell carcinoma and cervical adenocarcinoma, testicular germ cell tumor, thymoma, thyroid cancer, renal cell carcinoma, renal papillary cell carcinoma, adrenocortical carcinoma, pheochromocytoma, esophageal cancer, uterine carcinosarcoma, lung squamous cell carcinoma, and bladder urothelial carcinoma (CellSyst. 2018Mar28;6(3):282-300.e2).

[0063] In a particularly preferred embodiment of the present invention, MYCC gene amplification, MYCC gene mutation, or MYCC gene chromosomal location rearrangement may be used as a biomarker in ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer, and prostate cancer.

[0064] In another preferred embodiment of the present invention, the biomarkers may be MYCN gene amplification, MYCN gene mutation, or MYCN gene chromosomal location rearrangement in solid cancers selected from gastric cancer, lung cancer, breast cancer, hepatocellular carcinoma, head and neck squamous cell carcinoma, bile duct carcinoma, uveal melanoma, sarcoma, prostate cancer, colon cancer, cutaneous melanoma, mesothelioma, glioblastoma, glioma, cervical squamous cell carcinoma and cervical adenocarcinoma, testicular germ cell tumor, adrenocortical carcinoma, pheochromocytoma, esophageal cancer, uterine carcinosarcoma, lung squamous cell carcinoma, and bladder urothelial carcinoma (CellSyst. 2018Mar28;6(3):282-300.e2).

[0065] In a preferred embodiment of the present invention, the biomarkers may be MYCN gene amplification, MYCN gene mutation, or MYCN gene chromosomal location rearrangement in solid cancers selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, neuroblastoma, small cell lung cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, and retinoblastoma.

[0066] In a particularly preferred embodiment of the present invention, the biomarkers for neuroblastoma may be MYCN gene amplification, MYCN gene mutation, or MYCN gene chromosomal location rearrangement.

[0067] In another preferred embodiment of the present invention, the biomarkers may be MYCL gene amplification, MYCL gene mutation, or MYCL gene chromosomal location rearrangement in solid cancers selected from ovarian cancer, gastric adenocarcinoma, lung adenocarcinoma, breast cancer, hepatocellular carcinoma, head and neck squamous cell carcinoma, rectal adenocarcinoma, bile duct carcinoma, uveal melanoma, sarcoma, prostate adenocarcinoma, colon adenocarcinoma, cutaneous melanoma, mesothelioma, glioblastoma, glioma, cervical squamous cell carcinoma and adenocarcinoma, testicular germ cell tumor, thymoma, adrenocortical carcinoma, esophageal cancer, uterine carcinosarcoma, lung squamous cell carcinoma, and bladder urothelial carcinoma (Cell Syst. 2018 Mar 28; 6(3): 282-300.e2).

[0068] In a preferred embodiment of the present invention, the biomarkers for solid cancers selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer, and prostate cancer may be MYCL gene amplification, MYCL gene mutation, or MYCL gene chromosomal location rearrangement.

[0069] The biomarkers of the present invention can be used as biomarkers to predict the therapeutic response to therapy with said Compound A or its derivatives.

[0070] [Pharmaceutical Composition] The pharmaceutical composition of the present invention is a pharmaceutical composition for treating solid tumors having genetic characteristics predicted to confer sensitivity to sirtuin activating factor (CLK) inhibition, comprising compound A or a derivative thereof as an active ingredient.

[0071] The genetic feature may be selected from one or more of the following (a) to (k): (a) MYC gene amplification, (b) MYC gene mutation, (c) MYC gene chromosomal location rearrangement, (d) increased MYC mRNA level, (e) increased MYC protein level, (f) EGFR gene mutation, (g) U2AF1 mutation, (h) POLE gene loss, (i) CDK12 gene loss, (j) gene mutation selected from one or more of ATRX, ATR, GNAQ, and HIST1H3B, and (k) MDM4 exon skipping alteration. The MYC gene amplification, MYC gene mutation, or MYC gene chromosomal location rearrangement is detected by comparison with the respective references.

[0072] The MYC gene amplification may be observed to result in an increase in copy number compared to a reference, and may preferably, but not limited to, show an increase of about 1.5-fold or more. In other embodiments of the present invention, the increase may be, but is not limited to, an increase of about 2-fold or more, about 4-fold or more, about 10-fold or more, or about 20-fold or more.

[0073] The MYC gene mutation may be a mutation such as a single nucleotide variant (SNV), a multiple nucleotide variant (MNV), a small insertion or deletion (Indel), a splice site mutation, or a gene fusion within the genomic region of the MYC gene.

[0074] The MYC gene mutation may be, but is not limited to, preferably MYCC Q37del, P59L, S146L, MYCN R45Rfs*86 and / or E445K.

[0075] In a preferred embodiment of the present invention, the MYC gene may be selected from the group consisting of MYCC, MYCN, and MYCL.

[0076] The increase in the amount of MYC mRNA and / or MYC protein is detected by comparison with a reference.

[0077] The increase in the amount of MYC mRNA and / or the amount of MYC protein may be due to, but is not limited to, MYC gene amplification, MYC gene mutation, or MYC gene chromosomal site rearrangement.

[0078] The U2AF1 gene mutation is detected by comparison with a reference.

[0079] The U2AF1 gene mutation may be a mutation such as a single nucleotide variation (SNV), a multiple nucleotide variation (MNV), a small insertion or deletion (Indel), a splice site mutation, or a gene fusion within the genomic region of the gene, but is preferably an S34F mutation or an S34Y mutation.

[0080] The gene mutations selected from one or more of EGFR gene mutations, POLE gene loss, CDK12 gene loss, ATRX, ATR, GNAQ and HIST1H3B and / or MDM4 exon skipping changes are detected by comparison with the respective references.

[0081] The gene mutation selected from one or more of EGFR gene mutation, ATRX, ATR, GNAQ, and HIST1H3B and / or MDM4 exon skipping variation may each be a mutation such as a single nucleotide variation (SNV), a multiple nucleotide variation (MNV), a small insertion or deletion (Indel), a splice site mutation, or a gene fusion, and / or an RNA splicing variation.

[0082] The POLE gene reduction or CDK12 gene reduction may be any reduction that is observed in comparison with a reference.

[0083] The references used to detect the genetic characteristics of solid cancers may be obtained from non-cancerous tissue samples from the same subject or from databases well known in the art. The detection method may be any of those well known in the art, and is not limited thereto, but preferably, the above-mentioned detection methods can be used.

[0084] The solid tumors include cancers [e.g., colorectal cancer (e.g., colon cancer, rectal cancer, anal cancer, familial colorectal cancer, hereditary non-polyposis colorectal cancer, gastrointestinal stromal tumor), lung cancer (e.g., non-small cell lung cancer, small cell lung cancer, malignant mesothelioma), mesothelioma, pancreatic cancer (e.g., pancreatic ductal carcinoma, pancreatic endocrine tumor), pharyngeal cancer, laryngeal cancer, esophageal cancer, gastric cancer (e.g., papillary adenocarcinoma, mucinous adenocarcinoma, adenosquamous carcinoma), duodenal cancer, small intestinal cancer, breast cancer (e.g., invasive ductal carcinoma, ductal carcinoma in situ, inflammatory breast cancer), ovarian cancer (e.g., epithelial ovarian cancer, extragonadal germ cell tumor, ovarian germ cell tumor, ovarian low malignant potential tumor), testicular tumor, prostate cancer (e.g., hormone-dependent prostate cancer, hormone-independent prostate cancer, castration therapy-resistant prostate cancer), ), liver cancer (e.g., hepatocellular carcinoma, primary liver cancer, extrahepatic bile duct cancer), thyroid cancer (e.g., medullary thyroid carcinoma), kidney cancer (e.g., renal cell carcinoma (e.g., clear cell renal cell carcinoma), transitional cell carcinoma of the renal pelvis and ureter), uterine cancer (e.g., cervical cancer, uterine cancer, uterine sarcoma), gestational choriocarcinoma, brain tumor (e.g., medulloblastoma, glioma, pineal astrocytoma, pilocytic astrocytoma, diffuse astrocytoma, anaplastic astrocytoma, pituitary adenoma), retinoblastoma, skin cancer (e.g., basal cell carcinoma, malignant melanoma), sarcoma (e.g., rhabdomyosarcoma, leiomyosarcoma, soft tissue sarcoma, spindle cell sarcoma, clear cell sarcoma), malignant bone tumor, bladder cancer, cancer of unknown primary origin).

[0085] In a preferred embodiment of the present invention, the solid cancer may be selected from one or more of carcinoma, rhabdomyosarcoma, sarcoma such as, for example, alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fiber, blood vessels, adipose, and / or connective tissue), clear cell sarcoma, neuroblastoma, medulloblastoma, astrocytoma, glioblastoma multiforme, retinoblastoma, adenosquamous carcinoma, carcinosarcoma, mixed mesodermal tumor, teratocarcinoma, lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung carcinoma), breast cancer (including metastatic breast cancer), gastrointestinal cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney cancer or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

[0086] In a preferred embodiment of the present invention, the MYCC gene amplification, MYCC gene mutation, or MYCC gene chromosomal location rearrangement may be observed in ovarian cancer and solid cancers selected from gastric cancer, lung cancer, breast cancer, hepatocellular carcinoma, head and neck squamous cell carcinoma, rectal cancer, bile duct cancer, pancreatic cancer, uveal melanoma, sarcoma, prostate cancer, colon cancer, cutaneous melanoma, mesothelioma, glioblastoma, glioma, cervical squamous cell carcinoma and cervical adenocarcinoma, testicular germ cell tumor, thymoma, thyroid cancer, renal cell carcinoma, renal papillary cell carcinoma, adrenocortical carcinoma, pheochromocytoma, esophageal cancer, uterine carcinosarcoma, lung squamous cell carcinoma, and bladder urothelial carcinoma (CellSyst. 2018Mar28;6(3):282-300.e2).

[0087] In a particularly preferred embodiment of the present invention, the gene may be MYCC gene amplification, MYCC gene mutation, or MYCC gene chromosomal location rearrangement, which are found in ovarian cancer, breast cancer, stomach cancer, lung cancer, colon cancer, and prostate cancer.

[0088] In another preferred embodiment of the present invention, the MYCN gene amplification, MYCN gene mutation, or MYCN gene chromosomal location rearrangement may be observed in a solid cancer selected from gastric cancer, lung cancer, breast cancer, hepatocellular carcinoma, head and neck squamous cell carcinoma, bile duct carcinoma, uveal melanoma, sarcoma, prostate cancer, colon cancer, cutaneous melanoma, mesothelioma, glioblastoma, glioma, cervical squamous cell carcinoma and cervical adenocarcinoma, testicular germ cell tumor, acute myeloid leukemia, adrenocortical carcinoma, pheochromocytoma, esophageal cancer, uterine carcinosarcoma, lung squamous cell carcinoma, and bladder urothelial carcinoma (CellSyst. 2018Mar28;6(3):282-300.e2).

[0089] In a preferred embodiment of the present invention, the MYCN gene may be an amplification, a mutation, or a chromosomal rearrangement of the MYCN gene found in a solid cancer selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, neuroblastoma, small cell lung cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, and retinoblastoma.

[0090] In a particularly preferred embodiment of the present invention, the gene may be MYCN gene amplification, MYCN gene mutation, or MYCN gene chromosomal location rearrangement, which are found in neuroblastoma.

[0091] In another preferred embodiment of the present invention, the MYCL gene amplification, MYCL gene mutation, or MYCL gene chromosomal location rearrangement may be observed in ovarian cancer and solid cancers selected from gastric cancer, lung cancer, breast cancer, hepatocellular carcinoma, head and neck squamous cell carcinoma, rectal cancer, bile duct cancer, uveal melanoma, sarcoma, prostate cancer, colon cancer, cutaneous melanoma, mesothelioma, glioblastoma, glioma, cervical squamous cell carcinoma and cervical adenocarcinoma, testicular germ cell tumor, thymoma, adrenocortical carcinoma, esophageal cancer, uterine carcinosarcoma, lung squamous cell carcinoma, and bladder urothelial carcinoma (Cell Syst. 2018 Mar 28; 6(3): 282-300.e2).

[0092] In a preferred embodiment of the present invention, the MYCL gene may be an MYCL gene amplification, a MYCL gene mutation, or a MYCL gene chromosomal location rearrangement observed in a solid cancer selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer, and prostate cancer.

[0093] The pharmaceutical composition of the present invention can be used to treat solid cancers having the above-mentioned genetic characteristics. Furthermore, the pharmaceutical composition of the present invention can be administered to a subject in need of treatment for the solid cancer not only by administration alone but also in combination with other drugs. Furthermore, the pharmaceutical composition of the present invention may be used in combination with other therapies.

[0094] In this specification, each specific feature described in one embodiment relating to each aspect of the present invention may be combined in any manner to form a new embodiment, and it should be understood that such a new embodiment may also be included in each aspect of the present invention.

[0095] The present invention will be explained in more detail below with reference to examples, but the present invention is not limited to these examples in any way.

[0096] I. Drug sensitivity evaluation of Compound A (in vitro method) Example 1: Evaluation in various gene mutations This evaluation was carried out as follows based on the standard method of the MANO method described in WO 2018 / 181863.

[0097] (1) Cell Lines HEK (Human Embryonic Kidney) 293 cells and mouse 3T3 fibroblasts were obtained from the American Type Culture Collection (ATCC) and maintained in Dulbecco's modified Eagle's medium F-12 (DMEM-F12) supplemented with 10% FBS (both Thermo Fisher Scientific, Walthman, MA).

[0098] (2) Construction of Retroviral Vectors with Random Barcode Sequences A 60-bp fragment was inserted into the BamHI restriction enzyme site of the pcx4-bleo vector (https: / / www.addgene.org / vector-database / 2309 / ), followed by site-directed mutagenesis using a primer containing a 6-bp random sequence to create pcx5-belo, which contained a barcode sequence. Wild-type cDNA for each gene listed in Table 1 below was ligated to pcx5-belo. Furthermore, cDNA encoding each mutant was generated using the QuickChange II Site-Directed Mutagenesis Kit (Agilent Technologies, Santa Clara, CA) and ligated to pcx5-belo.

[0099] (3) Retrovirus Preparation and Cell Line Transduction The recombinant plasmid prepared as described above was co-transfected into HEK293 cells with a packaging plasmid (TakaraBio, Shiga, Japan) to obtain recombinant retroviral particles. For focus-forming assays, 3T3 cells were infected with ecotropic recombinant retroviruses using 4 μg / mL polybrene (Sigma-Aldrich, St. Louis, MO, USA) for 24 hours and then cultured in DMEM-F12 supplemented with 5% fetal bovine serum for up to two weeks. Cell transformation was assessed by phase-contrast microscopy or staining with Giemsa solution.

[0100] (4) Detection by MANO Method Genomic DNA derived from cell lysates was subjected to PCR using the primers 5'-TGCAAAGGACCTTACACAGTCCTG-3' and 5'-GACTCGTTGAAGGGTAGACTAGTC-3' to amplify the region containing the barcode sequence. The PCR conditions were as follows: Initial denaturation: 95°C for 3 minutes Denaturation: 95°C for 15 seconds Annealing: 60°C for 30 seconds Extension: 72°C for 60 seconds Number of cycles: 30 Final extension: 72°C for 10 minutes The amplified PCR product was purified using AMPurebeads (Beckman Coulter, Brea, CA). Sequencing libraries were prepared using the NEB Next Ultra DNA Library Prep Kit (NEB, Ipswich, MA) according to the manufacturer's recommendations. Library quality was assessed using a Qubit 2.0 Fluorometer (Thermo Fisher Scientific) and an Agilent 2200 Tape Station system. Libraries were sequenced using an Illumina Miseq (300 cycles) with Reagent Kit V2 to generate 150-bp paired-end reads. These sequence reads contained the following sequence, including the barcode sequence: 5'-CTAGACTGCCNNNNNNNGGATCACTCT-3' (where N is any nucleotide) and its complementary sequence. The amount of each variant was estimated by counting the reads of these barcode sequences. DMSO-treated cell mixtures were used as a reference control for scaling the signal of each cell clone. Therefore, the signal from each treated cell line was calculated as 100 × (median reads) / (median reads of DMSO control). Median reads are from three independent experiments.

[0101] (5) Results The signals based on the median read counts were summarized for each introduced gene (Figure 1). The IC90 or IC90 value of Compound A for genes with median read counts lower than those of GFP or KRASG12D (controls) are shown in Table 2 below.

[0102] From the above results, it was confirmed that Compound A exhibits a particularly strong growth inhibitory effect on cells having the above gene mutations.

[0103] Example 2: Evaluation of MYC gene mutations (1) Cell lines HEK (Human Embryonic Kidney) 293 cells and mouse 3T3 fibroblasts were obtained from ATCC (American Type Culture Collection) and maintained in Dulbecco's modified Eagle's medium F-12 (DMEM-F12) supplemented with 10% FBS (both Thermo Fisher Scientific, Walthman, MA).

[0104] (2) Construction of a retroviral vector having a random barcode sequence The vector used in Example 1 was used.

[0105] (3) Retrovirus Preparation and Cell Line Transduction The recombinant plasmid prepared as described above was co-transfected into HEK293 cells with a packaging plasmid (Takara Bio, Shiga, Japan) to obtain recombinant retroviral particles. For focus-forming assays, 3T3 cells were infected with ecotropic recombinant retroviruses using 4 μg / mL polybrene (Sigma-Aldrich, St. Louis, MO, USA) for 24 hours and then cultured in DMEM-F12 supplemented with 5% fetal bovine serum for up to two weeks. Cell transformation was assessed by phase-contrast microscopy or staining with Giemsa solution.

[0106] (4) Establishment of Cell Lines In vitro growth inhibition tests were performed using NIH3T3 cell lines established by individually introducing the MYCC WT, Q37del, P59L, S146L, MYCN WT, P45Rfs*86, E445K, or EGFR L858R genes found in the screening, or KRASG12D as a control.

[0107] (5) Cell Viability Assessment After seeding the four cell lines (1,000 cells / 90 μL) into each well of a 96-well plate, 10 μL of medium containing DMSO alone or Compound A (0.3 nM, 1 nM, 3 nM, 10 nM, 30 nM, 100 nM, 300 nM, 1000 nM, and 3000 nM) was quickly added to each well to achieve various final concentrations. After 3 days of culture, cell viability was assessed as follows. Cell Titer-Glo was used to measure intracellular adenosine triphosphate (ATP) levels to assess cell viability following each treatment. Specifically, 100 μL of Cell Titer-Glo solution was added per well, incubated for 15 minutes at room temperature, and then chemiluminescence was measured. The average value for each combination was calculated from the obtained data, and a concentration-response curve for cell viability for each drug alone or in combination was created, with the luminescence intensity of cells treated with DMSO alone set at 100%.

[0108] (6) Results The IC50 values ​​of NIH3T3 cell lines established by individual transfection of MYCC WT, Q37del, P59L, S146L, MYCN WT, P45Rfs*86, and E445K were lower than those of the NIH3T3 cell line established by transfection of KRASG12D as a control. These results confirmed that Compound A exhibits particularly strong growth inhibitory effects on cells overexpressing MYCC WT, Q37del, P59L, S146L, MYCN WT, P45Rfs*86, or E445K or having genetic abnormalities (Figure 2). On the other hand, the IC50 values ​​of the NIH3T3 cell lines established by individual transfection of EGFR L858R, MYCC S146L, and MYCN E445K were lower than that of the NIH3T3 cell line established by transfection of KRASG12D as a comparison. These results confirmed that compound A exhibits a particularly strong growth inhibitory effect on cells having the above gene mutations (Figure 3).

[0109] Example 3: Evaluation of MYCC gene amplification (1) Cell lines NCI-H82 cell line, AU565 cell line, NCI-H3122 cell line, and PC-9 cell line were obtained from ATCC (American Type Culture Collection) and maintained in Dulbecco's modified Eagle's medium F-12 (DMEM-F12) supplemented with 10% FBS (all Thermo Fisher Scientific, Waltham, MA).

[0110] (2) MYCC gene amplification in each cell line According to the Cancer Cell Line Encyclopedia database, the MYCC gene amplification in the NCI-H82 cell line and the AU565 cell line was 18.9 and 9.7, respectively. No gene amplification was confirmed in the NCI-H3122 cell line and the PC-9 cell line.

[0111] (3) Cell Viability Assessment. The two cell lines (1,000 cells / 90 μL) in which MYCC gene amplification was observed were seeded into each well of a 96-well plate. Then, 10 μL of medium containing DMSO alone or Compound A (1 nM, 3 nM, 10 nM, 30 nM, 100 nM, 300 nM, 1000 nM, 3000 nM, and 10,000 nM) was quickly added to each well to achieve various final concentrations. After 3 days of incubation, cell viability was assessed as follows. Cell Titer-Glo was used to measure intracellular adenosine triphosphate (ATP) levels to assess cell viability following each treatment. Specifically, 100 μL of Cell Titer-Glo solution was added to each well, incubated at room temperature for 15 minutes, and then chemiluminescence was measured. The average value for each combination was calculated from the obtained data, and the luminescence intensity in cells treated with DMSO alone was set as 100%, and a concentration-response curve of cell viability for each drug alone or in combination was prepared.

[0112] (4) Results The IC50 values ​​in the NCI-H82 and AU565 cell lines, which have MYCC gene amplification, were lower than those in the NCI-H3122 and PC-9 cell lines, which do not have gene amplification. These results confirmed that Compound A exhibits a particularly strong growth inhibitory effect on cells overexpressing MYCC (Figure 4).

[0113] Example 4: Evaluation of MYCN gene amplification (1) Cell culture Neuroblastoma cell lines CHP-212, SK-N-BE(2), SK-N-DZ, LN-18, U-87MG, and SW1088 were obtained from ATCC (American Type Culture Collection) and cultured in Dulbecco's modified Eagle's medium F-12 (DMEM-F12) supplemented with 10% FBS (all Thermo Fisher Scientific, Walthman, MA) under conditions of 5% CO2 and 37°C.

[0114] (2) Cell Viability Assessment After seeding the six cell lines described above (4,000 cells / 90 μL each) into each well of a 96-well plate, 10 μL of medium containing DMSO alone or Compound A (1 nM, 3 nM, 10 nM, 30 nM, 100 nM, 300 nM, 1000 nM, 3000 nM, and 10,000 nM) was quickly added to each well to achieve various final concentrations. After 4 days of culture, cell viability was assessed as follows. CellTiter-Glo was used to measure intracellular adenosine triphosphate (ATP) levels to assess cell viability following each treatment. Specifically, 100 μL of CellTiter-Glo solution was added to each well, incubated at room temperature for 15 minutes, and then chemiluminescence was measured. The average value for each combination was calculated from the obtained data, and the luminescence intensity in cells treated with DMSO alone was set as 100%, and a concentration-response curve of cell viability for each drug alone or in combination was prepared.

[0115] (3) Data Analysis The half maximal inhibitory concentration (IC50) for Compound A was determined using GraphPad Prism software version 7.0 (GRAPHPAD software).

[0116] (4) Results The evaluation results of IC50 for each cell line after treatment with Compound A are shown in Table 3. The IC50 values ​​for cell lines with MYCN gene amplification were lower than those for cell lines without MYCN gene amplification after treatment with Compound A. These results confirmed that Compound A exerts a stronger growth inhibitory effect on cell lines with MYCN gene amplification.

[0117] II. Evaluation of the antitumor effect of Compound A on mouse subcutaneous transplant models of various gene mutations (in vivo method) Example 5: Evaluation of MYCC gene amplification and the antitumor effect of Compound A in mouse subcutaneous transplant models of tumors derived from human cancer patients (1) Mouse subcutaneous transplant model Among the mouse subcutaneous transplant models of tumors derived from various human cancer patients established by the National Cancer Center Research Institute, this study used a total of seven models of five cancer types: ovarian cancer, lung cancer, gastric cancer, breast cancer, and colon cancer. Measurement results of MYCC gene amplification for these models have been obtained so far by the National Cancer Center Research Institute.

[0118] (2) Evaluation of antitumor activity Tumor pieces of 2 to 3 mm square were subcutaneously transplanted into the abdominal skin of 6 to 11 week old female NOG mice. The tumor volume was measured using a vernier caliper and calculated using the formula: tumor volume = (tumor major axis x tumor minor axis x tumor minor axis) / 2. When the tumor volume was approximately 140 to 230 mm ３ Once the tumor volume reached 100 mg / kg, the mice were divided into two groups with 2-4 mice per group. Compound A was orally administered at a dose of 25 mg / kg twice daily, twice weekly, for 14 or 28 days. A 0.5% methylcellulose solution was orally administered to the control group. After grouping, tumor volume was measured twice weekly. The T / C (%) on day 14 or 28 was used as an index of antitumor effect. T / C (%) = (change in tumor volume in drug-treated group) / (change in tumor volume in control group) x 100 (3) Results The results of evaluating the antitumor effect of Compound A in mouse subcutaneous transplant models of tumors derived from human cancer patients with five types of cancer (ovarian cancer, lung cancer, gastric cancer, breast cancer, and colon cancer) are shown in Table 4 below.

[0119] In the MYCC gene amplified model (MYCC CNV>2) used in this study, Compound A exhibited antitumor effects with T / C of -71.1%, -44.3%, -32.8%, and -28.0% (average -44.1%), whereas in the c-Myc gene unamplified model (MYCC CNV=2), Compound A exhibited antitumor effects with T / C of -27.8%, -26.1%, and -9.9% (average -21.3%).

[0120] From the above results, it was confirmed that Compound A exhibits a stronger antitumor effect in tumors with MYCC gene amplification.

[0121] From the above, it was demonstrated that MYCC gene amplification is useful as a biomarker for predicting the therapeutic response to Compound A.

[0122] Example 6: Evaluation of antitumor effect of compound A on mouse subcutaneous transplantation model of tumor derived from mouse with MMTV-MYCC or MMTV-HRAS (1) Evaluation of antitumor effect in mouse subcutaneous transplantation model Tumor pieces of mouse-derived tumor BTG-M605 with MMTV-MYCC or mouse-derived tumor BTG-R502 with MMTV-HRAS (both Shanghai Medicilon Inc.) were subcutaneously transplanted into the abdominal region of female nude mice. Tumor volume was measured using a vernier caliper and calculated using the formula: tumor volume = (tumor major axis x tumor minor axis x tumor minor axis) / 2. When the tumor volume was approximately 180 mm ３ Once the tumor volume reached 100 mg / kg, the mice were divided into four groups, each with 8 mice. Compound A was orally administered at doses of 12.5, 25, and 50 mg / kg twice daily, twice weekly (days 0, 4, 7, and 11) for 14 days. A 0.5% methylcellulose solution was orally administered to the control group. After grouping, tumor volume was measured twice weekly. The T / C (%) on day 14 was used as an index of antitumor effect. T / C (%) = (change in tumor volume in the drug-treated group) / (change in tumor volume in the control group) x 100. (2) Results The results of evaluating the antitumor effect of Compound A in mouse models with subcutaneously transplanted tumors BTG-M605, derived from a mouse with MMTV-MYCC, and BTG-R502, derived from a mouse with MMTV-HRAS, are shown in Tables 5 and 6, respectively. The T / C ratios on day 14 in the groups orally administered with compound A at doses of 12.5, 25, and 50 mg / kg were 29.1%, 19.6%, and 9.1%, respectively, in BTG-M605, and 71.1%, 54.4%, and 14.4%, respectively, in BTG-R502, demonstrating dose-dependent antitumor activity in both models. Furthermore, comparison of the results in the two models confirmed that compound A exerted a stronger antitumor effect in tumors with Myc gene amplification. Based on these findings, MYCC gene amplification is useful as a biomarker for predicting the therapeutic response to compound A.

[0123] Example 7: Evaluation of antitumor effect of compound A on a mouse subcutaneous transplantation model of a tumor from a human non-small cell lung cancer patient with a U2AF1 mutation (1) Evaluation of antitumor effect in a mouse subcutaneous transplantation model LU5231 (CrownBiosciences), a tumor from a human non-small cell lung cancer patient with an S34F mutation in the U2AF1 gene, was suspended in PBS:Cultrex ECM=1:1 and injected subcutaneously at 1.5 x 10 into the abdominal cavity of 6-8 week-old female NODSCID mice. ５ The tumor volume was measured using a vernier caliper and calculated using the formula: tumor volume = (tumor major axis x tumor minor axis x tumor minor axis) / 2. ３ When the tumor volume reached 1000 mg / kg, the mice were divided into three groups, with nine mice per group. Compound A was orally administered at doses of 6.25 and 12.5 mg / kg twice daily, twice weekly (days 1, 4, 8, 11, 15, and 18) for 21 days. A 0.5% methylcellulose solution was orally administered to the control group. After grouping, tumor volume was measured twice weekly. The T / C (%) on day 21 was used as an index of antitumor effect. Furthermore, human small cell lung cancer cell line NCI-H1048 (ATCC), which does not have a mutation in the U2AF1 gene, was suspended in HBSS:Matrigel = 1:1 and injected subcutaneously at 4 x 10 in the abdominal cavity of 6-week-old female nude mice. ６ The tumor volume was approximately 150 mm ３Once the tumor volume reached 100 mg / kg, the mice were divided into three groups, each with five mice. Compound A was orally administered at doses of 25 and 50 mg / kg twice daily, twice weekly (days 0, 3, 7, and 10) for 14 days. A 0.5% methylcellulose solution was orally administered to the control group. After grouping, tumor volume was measured twice weekly. The T / C (%) on day 14 was used as an index of antitumor effect. T / C (%) = (change in tumor volume in the drug-treated group) / (change in tumor volume in the control group) x 100. (2) Results The results of evaluating the antitumor effect of Compound A in a mouse subcutaneous transplant model of a human non-small cell lung cancer patient tumor LU5231 carrying an S34F mutation in the U2AF1 gene and a mouse subcutaneous transplant model of the human small cell lung cancer cell line NCI-H1048 carrying no S34F mutation in the U2AF1 gene are shown in Tables 7 and 8, respectively.

[0124] In the LU5231 model, the T / C ratios at day 21 for the groups orally administered compound A at doses of 6.25 and 12.5 mg / kg were 7.0% and -24.2%, respectively, demonstrating strong dose-dependent antitumor activity accompanied by tumor regression. Meanwhile, in the NCI-H1048 model, the T / C ratios at day 14 for the groups orally administered compound A at doses of 25 and 50 mg / kg were 35.0% and 6.2%, respectively. These results confirmed that compound A exerts a stronger antitumor effect in tumors harboring the S34F mutation in the U2AF1 gene compared to tumors lacking this mutation. These findings demonstrate that the S34F mutation in the U2AF1 gene is useful as a biomarker for predicting the therapeutic response to compound A.

[0125] III. Mutation Analysis of Cancer-Related Genes Example 8: Mutation Analysis of Cancer-Related Genes Using Oncomine Comprehensive Assay v3 In this analysis, tumor tissue samples were obtained from cases in which informed consent was obtained in a phase I clinical trial of Compound A for advanced, recurrent, or refractory malignant tumors, in accordance with a protocol approved by the Institutional Review Board of the National Cancer Center, and mutation analysis of cancer-related genes using Oncomine Comprehensive Assay v3 was performed according to the following procedure.

[0126] (1) Sequence Analysis Nucleic acid extraction was performed from the measurement samples using the RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE (Thermo Fisher Scientific). DNA was measured by fluorometric quantification using a Qubit™ 4 Fluorometer (Thermo Fisher Scientific) and qPCR quantification using a Step One Plus™ Real-Time PCR System (Thermo Fisher Scientific). RNA was measured by fluorometric quantification using the Qubit™ 4 Fluorometer alone, and nucleic acid concentration and amount were quantified. The extracted DNA and RNA were used to create a sequencing library using Oncomine™ Comprehensive Assay v3M (Thermo Fisher Scientific). The Ion Torrent™ NGS Reverse Transcription Kit (Thermo Fisher Scientific) was used for reverse transcription of the RNA library. Sequencing was performed using an Ion Gene Studio™ S5 Plus Sequencer (Thermo Fisher Scientific). From the data obtained by sequencing, mutations such as copy number variations (CNV), single nucleotide variations (SNV), multiple nucleotide variations (MNV), insertion / deletion (Indel), and exon skipping changes were identified using the Ion Reporter™ Server System (Thermo Fisher Scientific).

[0127] (2) Antitumor Effect of Compound A The best overall response for each case analyzed was partial response (PR) in two cases of ovarian cancer, stable disease (SD) in seven cases, and progressive disease (PD) in five cases. Of these 14 cases, three cases had a treatment period from the first day of administration to the last day of administration (treatment period) of more than 168 days (28 days / cycle x 6 cycles). Of the three cases, two were PR cases of ovarian cancer, and the other was SD case of ovarian cancer. It was believed that a strong therapeutic effect was observed in these three cases (Case A: best overall effect was SD, treatment period was 186 days; Case B: best overall effect was PR, treatment period was 193 days; Case C: best overall effect was PR, treatment period was 387 days), and the correlation with each mutation was analyzed.

[0128] (3) Correlation analysis with MYCC, POLE, and CDK12 CNV (i) MYCC CNV The MYCC CNVs of the three ovarian cancer cases in which a strong therapeutic effect was observed were 6.49 in Case A, 6.24 in Case B, and 4.99 in Case C. All three cases were above the threshold of 3, suggesting CNV amplification. Other than this, one case of transverse colon cancer (best overall response: SD, treatment period: 144 days) had a CNV amplification of 4.02. On the other hand, there was one ovarian cancer case in which a strong therapeutic effect was not observed (best overall response: PD, treatment period: 11 days), and the MYCC CNV in this case was 2.58, not exceeding the threshold of 3 or above. Similarly, none of the other cases in which a strong therapeutic effect was observed exceeded the threshold of 3, suggesting that MYCC CNV amplification may be able to predict the effectiveness of Compound A. Note that all of these CNVs were filtered as PASS or GAIN during data analysis, and are therefore considered to be highly accurate data.

[0129] (ii) POLE CNV The upper limit (95% point) of the 90% confidence interval of POLE CNV in the three ovarian cancer cases in which a strong therapeutic effect was observed was 0.84 in Case C, 1.47 in Case A, and 1.94 in Case B. Since all three cases were below the threshold of 2, it was considered that there was a decrease in CNV. On the other hand, in one case of ovarian cancer in which a strong therapeutic effect was not observed, the upper limit (95% point) of the 90% confidence interval was 3.01 (best overall effect was PD, treatment period was 11 days), which was not below the threshold of 2. Similarly, in the other 10 cases in which a strong therapeutic effect was not observed, none of the cases were below the threshold of 2, suggesting that a decrease in POLE CNV may be predictive of the effectiveness of Compound A. Note that all of these CNVs passed the filter during data analysis, and are therefore considered to be highly accurate data.

[0130] (iii) CDK12 CNV The upper limit (95% point) of the 90% confidence interval of CDK12 CNV in three cases of ovarian cancer in which a strong therapeutic effect was observed was 1.93 in Case A, 1.94 in Case C, and 1.98 in Case B. Since all three cases were below the threshold of 2, it was considered that there was a decrease in CNV. On the other hand, in one case of ovarian cancer in which a strong therapeutic effect was not observed, the upper limit (95% point) of the 90% confidence interval was 4.22 (best overall effect was PD, treatment period was 11 days), which was not below the threshold of 2. Similarly, in the other 10 cases in which a strong therapeutic effect was not observed, none of the cases were below the threshold of 2, suggesting that a decrease in CDK12 CNV may predict the effectiveness of compound A. Note that all of these CNVs passed the filter during data analysis, and are considered to be highly accurate data.

[0131] (iv) Clinical Significance These results suggest that the efficacy of Compound A, a sirtuin-activating factor (CLK) inhibitor, can be predicted by confirming MYCC CNV amplification, POLE CNV reduction, and CDK12 CNV reduction.

[0132] (4) Correlation analysis with SNVs of ATRX, ATR, GNAQ, and HIST1H3B (i) SNVs of ATRX, ATR, GNAQ, and HIST1H3B From the SNVs, MNVs, and indels of all cases, mutations that were thought to have a certain quality (selection was based on coverage, % frequency, and other quality indicators, and selection was also based on the influence of FFPE deamination in some specimens), cause changes in the amino acid sequence, and have a low minor allele frequency in East Asians were selected. Among these mutations, mutations present only in ovarian cancers that showed a strong therapeutic effect were p. Val936Leu of ATRX in case C, p. Thr2487Ala of ATR, p. Ala171Val of GNAQ, and p. Arg84Ser of HIST1H3B in case A. These results suggest that ATRX, ATR, GNAQ and / or HIST1H3B may have functions related to the efficacy of Compound A.

[0133] (ii) Clinical Significance These results suggest that impairment of the normal function of ATRX, ATR, GNAQ, and / or HIST1H3B due to SNV mutations or the like may enhance the efficacy of Compound A, a CLK inhibitor.

[0134] (5) Correlation analysis with MDM4 exon skipping (i) MDM4 exon skipping In MDM4 mRNA, the ratio of the number of sequence reads detecting the connection between Exon 5 and Exon 7 due to skipping of Exon 6 to the number of sequence reads detecting the connection between Exon 1 and Exon 2 was high in two of the three ovarian cancer cases in which a strong therapeutic effect was observed. The respective ratios were 0.163 in Case C and 0.127 in Case B, the first and second highest values ​​among the 14 cases examined.

[0135] (ii) Clinical Significance These results suggest that by identifying a variant of MDM4 in which Exon 6 is skipped, it may be possible to predict the efficacy of Compound A, a sirtuin-activating factor (CLK) inhibitor.

[0136] Example 9: Reanalysis of CNV using Oncomine Comprehensive Assay v3 In the CNV analysis in Example 8, correction was performed using the tumor fraction to remove the influence of non-tumor cells contaminating the sample, and the CNV of the tumor cell portion of the sample was estimated. Therefore, determining the tumor fraction with high accuracy is important for analyzing CNV of clinical tumor samples. In this reanalysis, the tumor fraction of each sample was obtained using an improved method from the same pathological sections used to obtain the tumor fraction in Example 8. Furthermore, these tumor fractions were used to reanalyze the sequence data obtained in Example 8, thereby obtaining a new, desirable CNV. The correlation between the CNV obtained in this reanalysis and the antitumor effect is shown below. Note that Cases A, B, and C are the same cases as Cases A, B, and C in Example 8, respectively.

[0137] (1) MYCC CNV The MYCC CNV of the three ovarian cancer cases in which a strong therapeutic effect was observed was 6.49 in Case A, 7.65 in Case B, and 5.41 in Case C. All three cases were above the threshold of 3, and therefore CNV amplification was considered to have occurred. Other than these, one case of transverse colon cancer (best overall effect was SD, number of days from the first day of administration to the last day of administration (treatment period) was 144 days) had a CNV of 4.02, and one case of thymoma (best overall effect was SD, treatment period was 99 days) had a CNV of 4.25. Although a strong therapeutic effect was not observed in these cases, these cases were above the threshold of 3, and therefore CNV amplification was considered to have occurred. On the other hand, there was one case of ovarian cancer in which a strong therapeutic effect was not observed (best overall effect was PD, treatment period was 11 days), and the MYCC CNV in that case was 2.58, which did not exceed the threshold of 3. Similarly, none of the other eight cases in which a strong therapeutic effect was not observed exceeded the threshold of 3. From the above results, in cases in which a strong therapeutic effect was observed, MYCC CNV tended to be amplified to a similar extent to or greater than that in Example 8, suggesting the possibility that MYCC CNV amplification may be able to predict the efficacy of Compound A. Note that all of these CNVs were filtered as PASS or GAIN during data analysis, and are considered to be highly accurate data.

[0138] (2) POLE CNV The upper limit (95% point) of the 90% confidence interval of POLE CNV in the three ovarian cancer cases in which a strong therapeutic effect was observed was 0.67 for Case C, 1.47 for Case A, and 1.91 for Case B. Since all three cases were below the threshold of 2, it was considered that there was a decrease in CNV. On the other hand, in one case of ovarian cancer in which a strong therapeutic effect was not observed, the upper limit (95% point) of the 90% confidence interval was 3.01 (best overall effect was PD, treatment period was 11 days), which was not below the threshold of 2. Similarly, in the other 10 cases in which a strong therapeutic effect was not observed, none of the cases were below the threshold of 2, suggesting that a decrease in POLE CNV may be predictive of the effectiveness of Compound A. Note that all of these CNVs passed the filter during data analysis, and are therefore considered to be highly accurate data.

[0139] (3) CDK12 CNV The upper limit (95% point) of the 90% confidence interval of CDK12 CNV in three cases of ovarian cancer in which a strong therapeutic effect was observed was 1.93 in Case A, 1.93 in Case C, and 1.97 in Case B. Since all three cases were below the threshold of 2, it was considered that there was a decrease in CNV. On the other hand, in one case of ovarian cancer in which a strong therapeutic effect was not observed, the upper limit (95% point) of the 90% confidence interval was 4.22 (best overall effect was PD, treatment period was 11 days), which was not below the threshold of 2. Similarly, in the other 10 cases in which a strong therapeutic effect was not observed, there was no case below the threshold of 2, suggesting that a decrease in CDK12 CNV may predict the effectiveness of compound A. Note that all of these CNVs passed the filter during data analysis, and are considered to be highly accurate data.

[0140] (4) Clinical Significance Based on these results, it was considered that in this reanalysis, similar to the CNV analysis in Example 8, it would be possible to predict the efficacy of Compound A, a sirtuin-activating factor (CLK) inhibitor, by confirming CNV amplification of MYCC, CNV reduction of POLE, and CNV reduction of CDK12.

[0141] Example 10: Additional analysis of mutations in cancer-related genes using Oncomine Comprehensive Assay v3 Mutation analysis of cancer-related genes using Oncomine Comprehensive Assay v3 was performed on four cases other than the 14 cases analyzed in Example 8. For these four cases, tumor tissue samples were obtained from cases in which informed consent was obtained in accordance with a protocol approved by the Institutional Review Board of the National Cancer Center Research Institute in a Phase I clinical trial of Compound A for advanced, recurrent, or refractory malignant tumors. Sequencing analysis was performed using the same method as in Example 8. However, the method used in Example 9 was used to calculate the tumor rate used in CNV analysis.

[0142] (1) Antitumor Effect of Compound A Of the four additional cases analyzed, two were administered only once, which was considered insufficient for analyzing the correlation with the antitumor effect of Compound A. Therefore, the remaining two cases were analyzed for the correlation with the antitumor effect of Compound A. The best overall response in these two cases was SD in one case and PD in one case. Of these, the number of days from the first day of administration to the last day of administration (treatment period) in one SD case exceeded 168 days (28 days / cycle x 6 cycles). This SD case, which was clear cell sarcoma (Case D: best overall response was SD, treatment period was 386 days), was considered to have demonstrated a strong therapeutic effect, and the correlation with each mutation was analyzed.

[0143] (2) Correlation analysis with MYCC, POLE, and CDK12 CNVs (i) MYCC CNVs The MYCC CNV of Case D, in which a strong therapeutic effect was observed, was 9.97, above the threshold of 3, suggesting a large CNV amplification. In the other case in which a strong therapeutic effect was not observed, the CNV did not exceed the threshold of 3. These results suggest that MYCC CNV amplification may be able to predict the efficacy of Compound A in this additional analysis. Note that these CNVs were PASS or GAIN in the filter during data analysis in both cases, and are therefore considered to be highly accurate data. (ii) POLE CNVs The upper limit (95% point) of the 90% confidence interval for POLE CNVs in Case D, in which a strong therapeutic effect was observed, and in the other case in which a strong therapeutic effect was not observed, was not less than the threshold of 2. This indicates that Compound A may be effective even without a decrease in POLE CNV, but this result does not seem to negate the possibility that Compound A, a sirtuin activating factor (CLK) inhibitor, may be effective if a decrease in POLE CNV occurs. Note that all of these CNVs passed the filter during data analysis, and are considered to be highly reliable data. (iii) CDK12 CNV The upper limit (95% point) of the 90% confidence interval for CDK12 CNV in Case D, in which a strong therapeutic effect was observed, and in the other case in which a strong therapeutic effect was not observed, was not less than the threshold value of 2. This indicates that Compound A may be effective even without a decrease in CDK12 CNV, but this result does not seem to negate the possibility that Compound A, a sirtuin activating factor (CLK) inhibitor, may be effective if a decrease in CDK12 CNV occurs. Note that all of these CNVs passed the filter during data analysis, and are considered to be highly reliable data. (iv) Clinical Significance From the results of Example 9 and this additional analysis, it was considered possible to predict the efficacy of Compound A, a sirtuin-activating factor (CLK) inhibitor, by confirming CNV amplification of MYCC, CNV reduction of POLE, and CNV reduction of CDK12.

[0144] (3) Correlation Analysis with MDM4 Exon Skipping Changes (i) MDM4 Exon Skipping Changes In MDM4 mRNA, the ratio of the number of sequence reads detecting the connection between Exon 5 and Exon 7 due to Exon 6 skipping to the number of sequence reads detecting the connection between Exon 1 and Exon 2 was not as high in Case D, in which a strong therapeutic effect was observed, and in the other case in which a strong therapeutic effect was not observed, as in the two cases in which a strong therapeutic effect was observed, as shown in Example 8. This indicates that Compound A may exhibit efficacy without a large value, but this result is not considered to negate the possibility that Compound A may exhibit efficacy if a large value were observed. (ii) Clinical Significance From the results of Example 8 and this additional analysis, it was considered possible to predict the efficacy of Compound A, a sirtuin-activating factor (CLK) inhibitor, by identifying a variant in which Exon 6 of MDM4 is skipped.

[0145] The biomarkers of the present invention are useful as biomarkers for predicting the therapeutic efficacy of CLK inhibitors such as Compound A in the treatment of solid cancers, including sarcomas.

Claims

1. In a subject with solid tumor, a compound of formula (I):

1. A method for predicting a therapeutic response to 1-((5-(1R)-1-fluoroethyl)-1,3,4-oxadiazol-yl)methyl)-6-(4-methoxypyrrolo[2,1-f][1,2,4]triazin-5-yl)-2-methyl-1H-imidazo[4,5-b]pyridine of Formula (I) or a derivative thereof, comprising: (1) detecting a genetic signature predicted to confer sensitivity to sirtuin-activating factor (CLK) inhibition in a sample derived from cancer tissue from the subject; and (2) determining the likelihood of the subject responding to the compound of Formula (I) or a derivative thereof based on the genetic signature.

2. The method of claim 1, wherein the genetic feature is one or more selected from the group consisting of: (a) MYC gene amplification, (b) MYC gene mutation, (c) MYC gene chromosomal rearrangement, (d) increased MYC mRNA level, (e) increased MYC protein level, (f) EGFR gene mutation, (g) U2AF1 gene mutation, (h) POLE gene loss, (i) CDK12 gene loss, (j) genetic mutation selected from one or more of ATRX, ATR, GNAQ, and HIST1H3B, and (k) MDM4 exon skipping change.

3. The method according to claim 2, comprising: (1) detecting MYC gene amplification, MYC gene mutation, MYC gene chromosomal location rearrangement, increased MYC mRNA level, and / or increased MYC protein level in a sample derived from cancer tissue from the subject; and (2) determining that the subject is particularly responsive to the compound represented by formula (I) or a derivative thereof when the MYC gene copy number is increased compared to a reference MYC gene copy number, the MYC gene sequence is altered compared to a reference MYC gene sequence, the MYC gene chromosomal location site is translocated compared to a reference MYC gene chromosomal location site, the MYC mRNA level is increased compared to the reference MYC mRNA level, and / or the MYC protein level is increased compared to the reference MYC protein level.

4. The method of claim 3, wherein the MYC gene is selected from MYCC, MYCN, and MYCL.

5. The method of claim 3, wherein the MYC gene copy number exhibits an increase of about 1.5-fold or more compared to the MYC gene copy number reference, or the MYC gene sequence exhibits MYCC Q37del, P59L, S146L, MYCN R45Rfs*86 and / or E445K compared to the reference.

6. The method of claim 3, comprising detecting the MYC gene amplification or MYC gene mutation by array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing, next generation sequencing (NGS), or multiplex ligation-dependent probe amplification (MLPA).

7. The method of claim 3, comprising detecting the rearrangement of the MYC gene chromosomal location site by in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next-generation sequencing (NGS), or comparative genomic hybridization (CGH).

8. The method of claim 3, comprising detecting the increased amount of MYC mRNA by in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next-generation sequencing (NGS), or real-time PCR or digital PCR analysis.

9. The method according to claim 3, comprising detecting the increased amount of MYC protein by immunohistochemical staining (IHC), flow cytometry (FACS), enzyme-linked immunosorbent assay (ELISA), proteinarray (Proximity Extension Assay, PEA method), or Western blotting (WB).

10. The method according to claim 2, comprising: (1) detecting, in a sample derived from cancer tissue from the subject, a gene mutation selected from one or more of EGFR gene mutation, U2AF1 gene mutation, POLE gene reduction, CDK12 gene reduction, ATRX, ATR, GNAQ, and HIST1H3B, and / or MDM4 exon skipping alteration; and (2) determining that the subject will be particularly responsive to the compound of formula (I) or a derivative thereof when a gene sequence selected from one or more of EGFR gene, U2AF1 gene, ATRX, ATR, GNAQ, and HIST1H3B is altered compared to a reference gene sequence of each gene, and / or when an MDM4 mRNA sequence is altered compared to a reference MDM4 mRNA sequence, and / or when the POLE gene copy number or CDK12 gene copy number is reduced compared to a reference copy number of each gene.

11. The method according to claim 10, wherein the U2AF1 gene mutation is an S34F mutation or an S34Y mutation.

12. The method of claim 10 or 11, comprising detecting the gene mutation selected from one or more of the EGFR gene mutation, the U2AF1 gene mutation, the POLE gene loss, the CDK12 gene loss, ATRX, ATR, GNAQ and HIST1H3B and / or the MDM4 exon skipping alteration by array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing or multiplex ligation-dependent probe amplification (MLPA), or RNA sequencing.

13. The method of any one of claims 1 to 12, wherein the cancer tissue-derived sample is a surgical sample, biopsy sample, pleural effusion sample, or ascites sample obtained from the subject, for example, from one or more of lung, blood, breast, gastrointestinal tract (stomach, colon, rectum), ovary, pancreas, liver, bladder, cervix, nerve, uterus, salivary gland, kidney, prostate, thyroid, or muscle tissue.

14. The method of any one of claims 1 to 12, wherein the subject is a human subject.

15.

13. The method of any one of claims 1 to 12, wherein the cancer is selected from one or more of carcinoma, sarcoma such as rhabdomyosarcoma, e.g., alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fibrous, blood vessels, adipose, and / or connective tissue), clear cell sarcoma, neuroblastoma, medulloblastoma, astrocytoma, glioblastoma multiforme, retinoblastoma, adenosquamous carcinoma, carcinosarcoma, mixed mesodermal tumor, teratocarcinoma, lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung carcinoma), breast cancer (including metastatic breast cancer), gastrointestinal cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney cancer or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

16. The method of any one of claims 3 to 9, wherein the MYC gene is MYCC and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer and / or prostate cancer.

17. The method of claim 16, wherein the MYC gene is MYCC and the cancer is ovarian cancer.

18. The method of any one of claims 3 to 9, wherein the MYC gene is MYCN and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer, prostate cancer, neuroblastoma, small cell lung cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme and / or retinoblastoma.

19. The method of claim 18, wherein the MYC gene is MYCN and the cancer is neuroblastoma.

20. The method of any one of claims 3 to 9, wherein the MYC gene is MYCL and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer and / or prostate cancer.

21. A compound of formula (I) below in a subject with solid tumor: a biomarker selected from one or more of the following (a) to (k) for predicting a therapeutic response to a therapy with 1-((5-(1R)-1-fluoroethyl)-1,3,4-oxadiazol-yl)methyl)-6-(4-methoxypyrrolo[2,1-f][1,2,4]triazin-5-yl)-2-methyl-1H-imidazo[4,5-b]pyridine or a derivative thereof, represented by the formula: (k) MDM4 exon skipping alteration.

22. The biomarker of claim 21, wherein the MYC gene is selected from MYCC, MYCN, and MYCL.

23. The biomarker according to claim 21, wherein the mutation in the U2AF1 gene is an S34F mutation or an S34Y mutation.

24. The biomarker of any one of claims 21 to 23, wherein the cancer is selected from the group consisting of carcinoma, rhabdomyosarcoma, sarcoma such as alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fibrous, vascular, adipose, and / or connective tissue), clear cell sarcoma, neuroblastoma, medulloblastoma, astrocytoma, glioblastoma multiforme, retinoblastoma, adenosquamous carcinoma, carcinosarcoma, mixed sarcoma, and the like. Biomarkers selected from one or more of germ layer tumors, teratocarcinoma, lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung cancer), breast cancer (including metastatic breast cancer), gastrointestinal cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

25. The biomarker of claim 21, wherein the MYC gene is MYCC and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer, and prostate cancer.

26. The biomarker of claim 25, wherein the MYC gene is MYCC and the cancer is ovarian cancer.

27. The biomarker of claim 21, wherein the MYC gene is MYCN and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, prostate cancer, neuroblastoma, small cell lung cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, and retinoblastoma.

28. The biomarker of claim 27, wherein the MYC gene is MYCN and the cancer is neuroblastoma.

29. The biomarker of claim 21, wherein the MYC gene is MYCL and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer, and prostate cancer.

30. A compound of formula (I):

1. A pharmaceutical composition comprising 1-((5-(1R)-1-fluoroethyl)-1,3,4-oxadiazol-yl)methyl)-6-(4-methoxypyrrolo[2,1-f][1,2,4]triazin-5-yl)-2-methyl-1H-imidazo[4,5-b]pyridine represented by the formula: or a derivative thereof as an active ingredient, wherein the gene characteristic is one or more selected from the following (a) to (k): (a) MYC gene amplification, (b) MYC gene mutation, (c) MYC gene chromosomal location rearrangement, (d) increased MYC mRNA amount, (e) increased MYC protein amount, (f) EGFR gene mutation, (g) U2AF1 gene mutation, (h) POLE gene reduction, (i) CDK12 gene reduction, (j) a genetic mutation selected from one or more of ATRX, ATR, GNAQ and HIST1H3B, and (k) an MDM4 exon skipping alteration.

31. The pharmaceutical composition of claim 30, wherein the MYC gene is selected from MYCC, MYCN, and MYCL.

32. The pharmaceutical composition according to claim 30, wherein the U2AF1 gene mutation is an S34F mutation or an S34Y mutation.

33. A pharmaceutical composition according to any one of claims 30 to 32, wherein the cancer is selected from the group consisting of carcinoma, rhabdomyosarcoma, sarcoma such as, for example, alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fiber, blood vessels, adipose, and / or connective tissue), clear cell sarcoma, neuroblastoma, medulloblastoma, astrocytoma, glioblastoma multiforme, retinoblastoma, adenosquamous carcinoma, carcinosarcoma, mixed sarcoma, and sarcoma. The pharmaceutical composition is selected from one or more of germ layer tumors, teratocarcinoma, lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung cancer), breast cancer (including metastatic breast cancer), gastrointestinal cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

34. The pharmaceutical composition of claim 30, wherein the MYC gene is MYCC and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer, and prostate cancer.

35. The pharmaceutical composition of claim 34, wherein the MYC gene is MYCC and the cancer is ovarian cancer.

36. The pharmaceutical composition of claim 30, wherein the MYC gene is MYCN and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, neuroblastoma, small cell lung cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, and retinoblastoma.

37. The pharmaceutical composition of claim 36, wherein the MYC gene is MYCN and the cancer is neuroblastoma.

38. The pharmaceutical composition according to claim 30, wherein the MYC gene is MYCL and the cancer is selected from ovarian cancer, breast cancer, gastric cancer, lung cancer, colon cancer, and prostate cancer.