Breast cancer treatment

Abstract

Disclosed is a therapeutic agent for breast cancer, said therapeutic agent comprising 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamide)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide or a pharmacologically acceptable salt thereof that is to be administered in combination with an estrogen receptor antagonist or an aromatase inhibitor.

Description

[Technical Field]

[0001] The present invention relates to a breast cancer therapeutic agent that combines a monocyclic pyridine derivative or a pharmacologically acceptable salt thereof having fibroblast growth factor receptor (FGFR) inhibitory activity with an estrogen receptor antagonist or an aromatase inhibitor. More specifically, the present invention relates to a breast cancer therapeutic agent that contains 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide or a pharmacologically acceptable salt thereof, administered in combination with an estrogen receptor antagonist or an aromatase inhibitor. [Background technology]

[0002] [ka]

[0003] 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) is known as an inhibitor of FGFR1, 2, and 3, and has been reported to have an inhibitory effect on cell proliferation of gastric cancer, lung cancer, bladder cancer, and endometrial cancer (Patent Document 1). The above compound has been reported to exhibit high therapeutic effects against bile duct cancer (Patent Document 2), breast cancer (Patent Document 3), and hepatocellular carcinoma (Patent Document 4). Succinate and maleate salts of the above compound are known as pharmacologically acceptable salts (Patent Document 5).

[0004] Estrogen receptor antagonists such as tamoxifen and fulvestrant are used to treat estrogen receptor-positive breast cancer (Non-Patent Document 1).

[0005] Aromatase inhibitors such as exemestane and anastrozole inhibit aromatase, which converts androgens secreted from the adrenal cortex into estrogens (Non-Patent Document 2). Therefore, like estrogen receptor antagonists, they are used to treat estrogen receptor-positive breast cancer. While estrogen receptor antagonists are generally used in premenopausal breast cancer patients, aromatase inhibitors are used in postmenopausal breast cancer patients.

[0006] Breast cancer is classified according to the presence or absence of expression of estrogen receptors, progesterone receptors, and human epidermal growth factor receptor type 2 (HER2), and in addition to surgery to remove the affected area, drug therapy is performed according to the type. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] U.S. Patent Application Publication No. 2014-0235614 [Patent Document 2] U.S. Patent Application Publication No. 2018-0015079 [Patent Document 3] U.S. Patent Application Publication No. 2018-0303817 [Patent Document 4] International Publication No. 2019 / 189241 [Patent Document 5] U.S. Patent Application Publication No. 2017-0217935 [Non-patent literature]

[0008] [Non-Patent Document 1] Howell et al., "Comparison of Fulvestrant Versus Tamoxifen for the Treatment of Advanced Breast Cancer in Postmenopausal Women Previously Untreated With Endocrine Therapy: A Multinational, Double-Blind, Randomized Trial", Journal of Clinical Oncology, 2004, 22(9), 1605-1613. [Non-patent document 2] Deeks et al.,"Exemestane A Review of its Use in Postmenopausal Women with Breast Cancer" Drugs, 2009, 69(7), 889-918 Summary of the Invention [Problem to be solved by the invention]

[0009] An object of the present invention is to provide a therapeutic agent for breast cancer that is administered by combining multiple drugs. [Means for solving the problem]

[0010] In view of this situation, the present inventors have conducted extensive research and have found that administering the compound represented by the above formula (I) in combination with an estrogen antagonist or an aromatase inhibitor exhibits a high therapeutic effect against breast cancer, thereby completing the present invention.

[0011] That is, the present invention provides the following [1] to

[18] . [1] A breast cancer therapeutic agent containing 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) or a pharmacologically acceptable salt thereof, characterized in that it is administered in combination with an estrogen receptor antagonist or an aromatase inhibitor. [ka] [2] A pharmaceutical composition for treating breast cancer, comprising 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) or a pharmacologically acceptable salt thereof, characterized in that it is administered in combination with an estrogen receptor antagonist or an aromatase inhibitor. [3] A pharmaceutical composition for treating breast cancer, comprising 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) or a pharmacologically acceptable salt thereof, and an estrogen receptor antagonist or an aromatase inhibitor: [4] A kit for treating breast cancer, comprising a formulation containing 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) or a pharmacologically acceptable salt thereof, and a formulation containing an estrogen receptor antagonist or an aromatase inhibitor. [5] A method for treating breast cancer, comprising administering 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) or a pharmacologically acceptable salt thereof, and an estrogen receptor antagonist or an aromatase inhibitor to a patient in need thereof. [6] 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) or a pharmacologically acceptable salt thereof for use in treating breast cancer, wherein the 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide is administered in combination with an estrogen receptor antagonist or an aromatase inhibitor. [7] A combination for treating breast cancer, comprising 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) or a pharmacologically acceptable salt thereof, and an estrogen receptor antagonist or an aromatase inhibitor: [8] Use of 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) or a pharmacologically acceptable salt thereof for the manufacture of a breast cancer therapeutic agent, characterized in that it is administered in combination with an estrogen receptor antagonist or an aromatase inhibitor. [9] The above-mentioned therapeutic agent, composition, kit, method, compound, combination, or use, wherein 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) or a pharmacologically acceptable salt thereof, and an estrogen receptor antagonist or an aromatase inhibitor are administered simultaneously or separately.

[10] The above therapeutic agent, composition, kit, method, compound, combination or use, wherein the pharmacologically acceptable salt is 1.5 succinate salt.

[11] The above-mentioned therapeutic agent, composition, kit, method, compound, combination or use, wherein the estrogen receptor antagonist is fulvestrant, tamoxifen or a pharmacologically acceptable salt thereof, or mepitiostane.

[12] The above-mentioned therapeutic agent, composition, kit, method, compound, combination or use, wherein the estrogen receptor antagonist is fulvestrant.

[13] The above-mentioned therapeutic agent, composition, kit, method, compound, combination or use, wherein the aromatase inhibitor is exemestane, anastrozole or letrozole.

[14] The above-mentioned therapeutic agent, composition, kit, method, compound, combination or use, wherein the aromatase inhibitor is exemestane. [14a] The above therapeutic agent, composition, kit, method, compound, combination or use, wherein the aromatase inhibitor is letrozole.

[15] The above-mentioned therapeutic agent, composition, kit, method, compound, combination or use, wherein the breast cancer is estrogen receptor positive.

[16] The above-mentioned therapeutic agent, composition, kit, method, compound, combination or use, wherein the breast cancer is locally advanced breast cancer, metastatic breast cancer, recurrent breast cancer or unresectable breast cancer.

[17] The above-mentioned therapeutic agent, composition, kit, method, compound, combination or use for treating breast cancer expressing fibroblast growth factor receptor (FGFR).

[18] The above therapeutic agent, composition, kit, method, compound, combination or use, wherein the FGFR is FGFR1, FGFR2 or FGFR3. [Effects of the Invention]

[0012] The administration of the compound represented by formula (I) in combination with an estrogen antagonist or an aromatase inhibitor may have the effect of reducing tumor volume in breast cancer. [Brief explanation of the drawings]

[0013] [Figure 1] 1 is a graph showing the change in average tumor volume over time in each group after the start of drug administration in Example 1. [Figure 2] 1 is a graph showing the change in average tumor volume over time in each group after the start of drug administration in Example 4. DETAILED DESCRIPTION OF THE INVENTION

[0014] The compound represented by formula (I) according to the present invention or a pharmacologically acceptable salt thereof can be produced by the method described in Patent Document 1.

[0015] In this specification, pharmacologically acceptable salts include, for example, salts with inorganic acids, salts with organic acids, and salts with acidic amino acids.

[0016] Suitable examples of salts with inorganic acids include salts with hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.

[0017] Suitable examples of salts with organic acids include salts with acetic acid, succinic acid, fumaric acid, maleic acid, tartaric acid, citric acid, lactic acid, stearic acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, and p-toluenesulfonic acid.

[0018] Suitable examples of salts with acidic amino acids include salts with aspartic acid, glutamic acid, and the like.

[0019] The preferred pharmacologically acceptable salt is succinate or maleate, and the more preferred salt is succinate, particularly 1.5 succinate (hereinafter, the 1.5 succinate of the compound represented by formula (I) will be referred to as Compound A).

[0020] The breast cancer therapeutic agent of the present invention can be orally administered in the form of a solid preparation such as tablets, granules, fine granules, powder, capsules, etc., or in the form of a liquid preparation, jelly, syrup, etc. The breast cancer therapeutic agent of the present invention may also be administered parenterally in the form of an injection, suppository, ointment, cataplasm, etc.

[0021] The breast cancer therapeutic agent of the present invention can be formulated by the methods described in the Japanese Pharmacopoeia (JP), the European Pharmacopoeia (EP), or the United States Pharmacopoeia (USP).

[0022] The dosage of the compound represented by formula (I) or a pharmacologically acceptable salt thereof can be appropriately selected depending on the severity of symptoms, the patient's age, sex, body weight, sensitivity, administration method, administration timing, administration interval, dosage form, etc. When orally administered to an adult (body weight 60 kg), the daily dosage is 0.5 mg to 5 g, preferably 1 mg to 1 g, and more preferably 1 mg to 500 mg. This can be administered in 1 to 3 divided doses per day.

[0023] As used herein, an estrogen receptor antagonist refers to a drug that binds to the estrogen receptor expressed in breast cancer cells, thereby inhibiting the binding of estrogen to the estrogen receptor and suppressing the proliferation of breast cancer cells.

[0024] Suitable examples of estrogen receptor antagonists include fulvestrant, tamoxifen or a pharmacologically acceptable salt thereof (such as citrate), mepitiostane, etc. Preferred is fulvestrant.

[0025] [ka]

[0026] The estrogen receptor antagonist may also be elacestrant, H3B-6545, toremifene citrate, SAR439859, AZD9833, lintodestrant, ZN-c5, LSZ102, D-0502, LY3484356, SHR9549, brillanestran, or giredestrant.

[0027] The dosage and method of administration of the estrogen receptor antagonist, for example, in the case of fulvestrant, is 500 mg administered intramuscularly initially, two weeks later, four weeks later, and then every four weeks thereafter.

[0028] When the estrogen receptor antagonist is tamoxifen or a pharmacologically acceptable salt thereof, 20 to 40 mg (as tamoxifen) per day is orally administered in one to two divided doses.

[0029] When the estrogen receptor antagonist is mepitiostane, the recommended oral dose is 20 mg per day in two divided doses.

[0030] Alternatively, instead of an estrogen receptor antagonist, an aromatase inhibitor that inhibits estrogen synthesis may be used. Suitable examples of aromatase inhibitors include exemestane, anastrozole, letrozole, etc., with exemestane being preferred.

[0031] [ka]

[0032] The dosage and method of administration of aromatase inhibitors are, for example, 25 mg of exemestane, administered orally once a day after a meal.

[0033] If the aromatase inhibitor is anastrozole, it is administered orally at a dose of 1 mg once daily.

[0034] If the aromatase inhibitor is letrozole, administer 2.5 mg orally once daily.

[0035] As used herein, "administering in combination" means administering to a patient a formulation containing the compound represented by formula (I) or a pharmacologically acceptable salt thereof and a formulation containing an estrogen receptor antagonist or an aromatase inhibitor simultaneously or separately. Alternatively, a composition containing the compound represented by formula (I) or a pharmacologically acceptable salt thereof and an estrogen receptor antagonist or an aromatase inhibitor in a single formulation may be administered.

[0036] As used herein, breast cancer refers to benign or malignant tumors arising in the mammary glands (ducts and lobules), including locally advanced breast cancer, metastatic breast cancer, recurrent breast cancer, and unresectable breast cancer. [Example]

[0037] The present invention will be described in more detail below with reference to examples.

[0038] Example 1 Growth inhibitory effect of the combined use of Compound A and fulvestrant (trade name: Faslodex intramuscular injection 250 mg, AstraZeneca K.K.) on tumors (OD-BRE-0438) derived from human breast cancer patients The antitumor effects of Compound A and fulvestrant were evaluated using five NOD-SCID mice per group (NOD.CB17-Prkdcscid / J, female, Charles River Japan). OD-BRE-0438 is a hormone receptor-positive breast cancer patient-derived tumor established by Oncodesign. Tumor fragments were subcutaneously transplanted into NOD-SCID mice for passage. The tumors excised from the mice were cut into approximately 5 mm cubes and transplanted into the subcutaneous tissue of the right flank of each mouse using a trocar (Φ3.5 mm) for evaluation of antitumor effects. β-estradiol (Fujifilm Wako Pure Chemical Industries, Ltd.) was dissolved in 99.5% ethanol (Fujifilm Wako Pure Chemical Industries, Ltd.) at a concentration of 1 mg / mL, and then diluted with sterile water for drinking water to a final concentration of 2.5 μg / mL. This solution was administered to mice via drinking water from the day of tumor implantation until the end of the study. The major and minor diameters of the tumor were measured using an electronic digital caliper (Digimatic TM The tumor volume was measured using a caliper (Mitutoyo Corporation). The tumor volume was calculated according to the following formula. Tumor volume (mm 3 ) = major diameter (mm) × minor diameter (mm) × minor diameter (mm) / 2

[0039] Compound A was dissolved in purified water to a concentration of 2.5 mg / mL. As fulvestrant, a commercially available formulation (50 mg / mL) was used as is. Thirty days after tumor transplantation, the mice were assigned to each group so that the mean tumor volume was the same.

[0040] The first day of administration was designated as day 0, and the drugs were administered under the following conditions. Mice in each group were orally administered Compound A at a dose of 25 mg / kg (10 mL / kg) once daily for 14 consecutive days, and fulvestrant was subcutaneously injected at a dose of 250 mg / kg (5 mL / kg) on ​​days 0 and 7. The control group was untreated.

[0041] The tumor volume of each mouse was measured on days 0, 4, 7, 11, and 14. The results (mean values) are shown in Table 1 and Figure 1. Dunnett's multiple test was performed on the tumor volumes of the control group and each treatment group on day 14.

[0042] [Table 1]

[0043] Reference Example 1: Construction of a human CYP19A1 expression vector The PB-CMV-MCS-EF1α-Puro vector (System Biosciences) was digested with Xba I and Not I, and a multiple cloning site was inserted into it to create the PB510B2 vector. Next, the PB510B2 vector was digested with Xba I and Cla I, and the ORF of human CYP19A1 (NM_001347249.2) was inserted to obtain the PB510B2_hCYP19A1 vector.

[0044] Reference Example 2: Construction of pC3-PBase vector The neomycin expression module (SV40 promoter-Neomycin ORF-SV40 polyA) of the pcDNA3.1(-) Mammalian Expression Vector (Thermo Fisher Scientific) was deleted to construct the pC3-vector. Next, the Super PiggyBac Transposase ORF of the Super PiggyBac Transposase Expression Vector (System Biosciences) was inserted downstream of the CMV promoter of the pC3-vector to obtain the pC3-PBase vector.

[0045] Reference Example 3 Establishment of human-derived breast cancer cell line ZR-75-1-hCYP19A1 Human breast cancer cell line ZR-75-1 (ECACC) was seeded onto a 6-well microplate (FALCON). The culture medium used was RPMI-1640 medium (Fujifilm Wako Pure Chemical Industries, Ltd.) containing 10% FBS (SIGMA) and penicillin / streptomycin (Fujifilm Wako Pure Chemical Industries, Ltd.) (containing 4,500 mg / L glucose, L-glutamine, phenol red, HEPES, and sodium pyruvate). The seeded cells were cultured for 15 days in an incubator at 37°C and 5% CO2.

[0046] Lipofectamine TM 3000 Reagent (Thermo Fisher Scientific) (3.75 μL), Opti-MEM TM Solution A was prepared by mixing 125 μL of the PB510B2_hCYP19A1 vector (2 μg), pC3-PBase (0.5 μg), P3000 Reagent (Thermo Fisher Scientific) (5 μL), and Opti-MEM (Thermo Fisher Scientific). TM(125 μL) were mixed to prepare solution B. Solutions A and B were mixed and left to stand at room temperature for 5 minutes, then added to the ZR-75-1 cells obtained by the above culture, and cultured overnight at 5% CO2 and 37°C. The cells were then cultured in medium supplemented with 1 μg / mL puromycin.

[0047] Reference Example 4 Establishment of human-derived breast cancer cell line MCF-7-hCYP19A1 Human breast cancer cell line MCF-7 (ECACC) was seeded onto a 6-well microplate (Falcon). The culture medium used was E-MEM medium (Fujifilm Wako Pure Chemical Industries, Ltd.) supplemented with 10% FBS (Sigma), penicillin / streptomycin (Fujifilm Wako Pure Chemical Industries, Ltd.), and L-glutamine, phenol red, sodium pyruvate, non-essential amino acids, and 1500 mg / L sodium bicarbonate. The seeded cells were cultured overnight in an incubator at 37°C with 5% CO2.

[0048] Lipofectamine TM 3000 Reagent (3.75 μL), Opti-MEM TM Solution A was prepared by mixing the PB510B2_hCYP19A1 vector (2 μg), pC3-PBase (0.5 μg), P3000 reagent (5 μL), and Opti-MEM (125 μL). TM Solution A and solution B were mixed and left to stand at room temperature for 5 minutes, then added to the MCF-7 cells obtained by the above culture. The cells were then cultured for 3 days under conditions of 5% CO2 and 37°C. After that, the cells were cultured for 7 days in medium supplemented with 1 μg / mL puromycin, and then in medium supplemented with 1.5 μg / mL puromycin.

[0049] Example 2 Growth inhibitory effect of the combined use of Compound A and exemestane on subcutaneously transplanted tumors of human breast cancer cell line ZR-75-1-hCYP19A1 Five BALB / c nude mice (CAnN.Cg-Foxn1 / CrlCrlj, female, Charles River, Japan) were used in each group to evaluate the antitumor effects of Compound A and exemestane.

[0050] Mice were subcutaneously injected with a mixture of 0.3 mg / kg medetomidine hydrochloride, 4 mg / kg midazolam, and 5 mg / kg butorphanol tartrate. The dorsal skin and peritoneum were incised, the left and right ovaries were removed, and the incision was sutured. Subsequently, 65 mg / kg (13.0 mg / mL, 100 μL / mouse) ampicillin sodium injection (Meiji Seika Pharma), 1 mg / kg (0.2 mg / mL, 100 μL / mouse) antisedan (Nippon Zenyaku Kogyo), and 5 mg / kg (1.0 mg / mL, 100 μL / animal) carprofen injection (Zoetis Japan) were subcutaneously administered.

[0051] Seven days later, androstenedione (Sigma-Aldrich) was orally administered to the mice at 0.1 mg / mouse (200 μL / mouse) until the end of the test.

[0052] The day after starting androstenedione administration, 1 × 10 7 The human breast cancer cell line ZR-75-1-hCYP19A1 was implanted subcutaneously on the right side of the mice. 20 days after implantation, the mice were assigned to each group so that the average tumor volume was the same.

[0053] Compound A was dissolved in purified water to a concentration of 2.5 mg / mL. Tween (registered trademark) 80 was mixed with a 0.5% methylcellulose solution to a concentration of 0.4%. Exemestane was dissolved in this mixture to a concentration of 5 mg / mL.

[0054] The administration start date was designated as day 0, and the drugs were administered under the following conditions. Mice in each group were orally administered compound A at 50 mg / kg (20 mL / kg), exemestane at 1 mg / mouse (200 μL / mouse), or both compound A and exemestane (each at the same dose as above) once daily for 11 consecutive days. The control group was untreated.

[0055] The tumor volume of each mouse was measured on days 0, 4, 7, and 11. The results (mean values) are shown in Table 2. The major and minor diameters of the tumor were measured using an electronic digital caliper (Digimatic TM The tumor volume was measured using a caliper (Mitutoyo Corporation). The tumor volume was calculated according to the following formula. Tumor volume (mm 3 ) = major diameter (mm) × minor diameter (mm) × minor diameter (mm) / 2

[0056] [Table 2]

[0057] Example 3 Effect of the combination of Compound A and letrozole, anastrozole, or exemestane using human aromatase-expressing human-derived breast cancer cell line MCF-7-hCYP19A1 The human breast cancer cell line MCF-7-hCYP19A1 prepared in Reference Example 4 was cultured and maintained in E-MEM medium (containing L-glutamine, phenol red, sodium pyruvate, non-essential amino acids, and 1500 mg / L sodium bicarbonate, Fujifilm Wako Pure Chemical Industries) containing 10% FBS (SIGMA), penicillin / streptomycin (Fujifilm Wako Pure Chemical Industries), and 1.5 μg / mL puromycin (hereinafter referred to as culture medium) in a 5% CO2 incubator.

[0058] To each well of a cell culture plate, add 0.25 × 10 cells in culture medium containing 10% FBS. 4 200 μL of each cell suspension prepared at 100 cells / mL was added to each well and cultured overnight in an incubator.

[0059] The next day, the medium was removed, and 100 μL of phenol red-free RPMI-1640 medium (Fujifilm Wako Pure Chemical Industries, Ltd.) containing 10% FBS (Charcoal-Stripped FBS, GIBCO), penicillin / streptomycin, and 1.5 μg / mL puromycin (hereafter referred to as assay medium) was added, followed by incubation in an incubator.

[0060] The next day, the medium was removed, and 100 μL of assay medium was added to each well. The cells were then cultured in an incubator for another 2 days. The medium was then removed, and 25 μL of androstenedione (10 nmol / L in assay medium), 25 μL of FGF2 (GIBCO) (25 ng / mL in assay medium), and 25 μL of FGF10 (R&D Systems) (100 ng / mL in assay medium) were added. Then, 25 μL of each of the solutions listed below was added, and the cells were cultured in an incubator for 7 days. On days 3 and 5 after drug addition, the medium was removed, and 25 μL of androstenedione, FGF2, and FGF10 solutions, as well as each of the solutions listed below, were added for a medium change. (1) Assay medium containing letrozole (SIGMA) adjusted to a final concentration of 2000 nmol / L, anastrozole (SIGMA) adjusted to a final concentration of 10,000 or 2000 nmol / L, exemestane (SIGMA) adjusted to a final concentration of 2000 nmol / L or 400 nmol / L, or a final concentration of 0.05% DMSO (used as a control group or for Compound A alone) (2) Assay medium containing Compound A adjusted to a final concentration of 1000 nmol / L in assay medium or DMSO at a final concentration of 0.005% (used as a control group or in the case of a single drug described in (1)).

[0061] After culturing, the number of cells in each well was counted using CellTiter Glo TM 2.0 (Promega), the luminescence intensity was measured, and the amount of ATP contained in the cells was calculated.

[0062] After removing the medium from the plate, 50 μL of a 1:1 mixture of the measurement reagent and culture medium was added to each well. The plate was mixed for 5 minutes using a plate mixer, and then the plate was read using a plate reader (EnVISION TM Luminescence was measured using a luminescence analyzer (PerkinElmer). The luminescence value of each well was subtracted from the luminescence value of the wells containing no cells, and the average values ​​(n=3) for each group are shown in Tables 3 to 7.

[0063] [Table 3]

[0064] [Table 4]

[0065] [Table 5]

[0066] [Table 6]

[0067] [Table 7]

[0068] Example 4 Growth inhibitory effect of the combined use of Compound A and Letrozole on subcutaneously transplanted tumors of human breast cancer cell line MCF-7-hCYP19A1 Five BALB / c nude mice (CAnN.Cg-Foxn1 / CrlCrlj, female, Charles River, Japan) were used in each group to evaluate the antitumor effect of Compound A and letrozole.

[0069] Mice were subcutaneously injected with a mixture of 0.3 mg / kg medetomidine hydrochloride, 4 mg / kg midazolam, and 5 mg / kg butorphanol tartrate. The dorsal skin and peritoneum of the mice were incised, the left and right ovaries were removed, and the incision was sutured. Subsequently, Baytril 2.5% injection (Bayer) diluted with saline (Otsuka Pharmaceutical Factory) to a final concentration of 1 mg / mL enrofloxacin was administered subcutaneously at a dose equivalent to 5 mg / kg (100 μL / mouse). Antisedan (Nippon Zenyaku Kogyo) diluted with saline (Otsuka Pharmaceutical Factory) to a final concentration of 0.2 mg / mL atibamezole hydrochloride was administered subcutaneously at a dose equivalent to 1 mg / kg (100 μL / mouse).

[0070] Seven days later, testosterone enanthate (Fuji Pharma Co., Ltd.) was diluted with sesame oil to a concentration of 15.6 mg / mL and administered intramuscularly at 0.1 mL / mouse (1.56 mg / mouse). Thereafter, the mice were administered intramuscularly once a week until the end of the study.

[0071] The day after starting testosterone enanthate administration, 1 × 10 7 The human breast cancer cell line MCF-7-hCYP19A1 was implanted subcutaneously on the right side of the mice. Nine days after implantation, the mice were assigned to each group so that the average tumor volume was the same.

[0072] Compound A was dissolved in purified water to a concentration of 2.5 mg / mL, and letrozole was dissolved in 0.3% hydroxypropyl cellulose / sterilized water to a concentration of 0.05 mg / mL.

[0073] The administration start date was designated as day 0, and the drugs were administered under the following conditions. Mice in each group were orally administered compound A at 25 mg / kg (10 mL / kg), letrozole at 0.01 mg / mouse (200 μL / mouse), or both compound A and letrozole (same doses as above) once daily for 14 consecutive days. The control group was untreated.

[0074] The tumor volume of each mouse was measured on days 0, 4, 7, 11, and 14. The results (mean values) are shown in Table 8 and Figure 2. The major and minor diameters of the tumor were measured using an electronic digital caliper (Digimatic™ caliper, Mitutoyo Corporation). The tumor volume was calculated according to the following formula: Tumor volume (mm 3 ) = major diameter (mm) × minor diameter (mm) × minor diameter (mm) / 2 Dunnett's multiple range test was performed on the tumor volumes of the control group and each treatment group on day 14.

[0075] [Table 8]

Claims

1. A breast cancer therapeutic agent containing 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) or a pharmacologically acceptable salt thereof, characterized in that it is administered in combination with fulvestrant or elacestrant. 【Chemistry 1】

2. 2. The therapeutic agent according to claim 1, wherein 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I) or a pharmacologically acceptable salt thereof, and fulvestrant or elacestrant are administered simultaneously or separately.

3. 3. The therapeutic agent according to claim 1, wherein the pharmacologically acceptable salt is 1.5 succinate.

4. A therapeutic agent described in any one of claims 1 to 3, wherein the fulvestrant or elacestrant is fulvestrant.

5. A therapeutic agent described in any one of claims 1 to 3, wherein the fulvestrant or elacestrant is elacestrant.

6. The therapeutic agent according to any one of claims 1 to 5, wherein the breast cancer is estrogen receptor positive.

7. The therapeutic agent according to any one of claims 1 to 6, wherein the breast cancer is locally advanced breast cancer, metastatic breast cancer, recurrent breast cancer, or unresectable breast cancer.

8. The therapeutic agent according to any one of claims 1 to 7, which is for treating breast cancer expressing fibroblast growth factor receptor (FGFR).

9. The therapeutic agent according to claim 8, wherein the FGFR is FGFR1, FGFR2 or FGFR3.

10. A pharmaceutical composition for treating breast cancer containing 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide or a pharmacologically acceptable salt thereof, characterized in that it is administered in combination with fulvestrant or elacestrant.

11. A pharmaceutical composition for treating breast cancer, comprising 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide or a pharmacologically acceptable salt thereof, and fulvestrant or elacestrant.

12. 12. The pharmaceutical composition of claim 10 or 11, wherein the fulvestrant or elacestrant is fulvestrant.

13. 12. The pharmaceutical composition of claim 10 or 11, wherein the fulvestrant or elacestrant is elacestrant.

14. A kit for treating breast cancer, comprising a formulation containing 5-((2-(4-(1-(2-hydroxyethyl)piperidin-4-yl)benzamido)pyridin-4-yl)oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide or a pharmacologically acceptable salt thereof, and a formulation containing fulvestrant or elacestrant.

15. 15. The kit of claim 14, wherein the fulvestrant or elacestrant is fulvestrant.

16. 15. The kit of claim 14, wherein the fulvestrant or elacestrant is elacestrant.

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