Antitumor agent, antitumor effect enhancer, and antitumor kit

Abstract

The present invention aims to provide an antitumor agent, an antitumor kit and an antitumor effect enhancer having excellent antitumor effects compared to gemcitabine, an antitumor platinum complex and their combined therapies. According to the present invention, an antitumor agent comprising an antitumor platinum complex and 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof is provided.

Description

Technical Field

[0001] The present invention relates to an antitumor agent, an antitumor effect enhancer and an antitumor kit. Background Art

[0002] It is known that 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine (hereinafter sometimes referred to as "Compound A") has excellent antitumor activity and is useful as an antitumor agent (Patent Document 1). In addition, it is known that Compound A also has strong antitumor activity when orally administered to mice (Non-Patent Documents 1 and 2). In addition, salts of Compound A and methods for preparing the same are also known (Patent Documents 2 to 4). Furthermore, it is known that there are antitumor drugs that enhance the antitumor effect by combining specific acylthiourea compounds and antitumor agents (paclitaxel, gemcitabine, lapatinib, tegafur / gimeracil / oteracil potassium combination, irinotecan, etc.) (Patent Document 5).

[0003] In cancer chemotherapy, as a representative agent of antitumor agent, antitumor platinum complex is known. And, as platinum preparation (platinum preparation), it is known to have a preparation comprising antitumor platinum complex as an effective ingredient. As representative antitumor platinum complex, cisplatin, carboplatin, oxaliplatin and nedaplatin etc. can be enumerated, which are used to treat various malignant tumors. However, in clinical practice, it is easy to cause high nephrotoxicity and anticancer property to become a problem (non-patent literature 3).

[0004] In clinical practice, multiple-dose combination therapy is performed for the purpose of compensating for the difference in sensitivity of each antitumor agent to tumors and enhancing the efficacy, and antitumor platinum complexes are often used in combination therapy with other antitumor agents. In particular, as examples, combination therapy with taxanes such as paclitaxel and docetaxel, camptothecins such as irinotecan and topotecan, gemcitabine, 5-fluorouracil and capecitabine, nucleic acid metabolism antagonists such as tegafur / gimeracil / oteracil calcium combination (S-1), topoisomerase II inhibitors such as etoposide and doxorubicin, etc. is known, and it is used as a therapeutic agent for a wide range of solid cancers.

[0005] For example, the efficacy of the combined use of gemcitabine and cisplatin for pancreatic cancer patients was 11.5% and the median survival time was 7.5 months (Non-Patent Document 4), and the median survival time was 5.7 months and the efficacy was 9% (Non-Patent Document 5). Regardless of the combination therapy with any antitumor platinum complex, the therapeutic effect is not sufficiently high.

[0006] Prior art literature

[0007] Patent Literature

[0008] Patent Document 1: International Publication No. 1997 / 038001

[0009] Patent Document 2: International Publication No. 2013 / 146833 Pamphlet

[0010] Patent Document 3: International Publication No. 2011 / 074484 Pamphlet

[0011] Patent Document 4: International Publication No. 2014 / 027658 Pamphlet

[0012] Patent Document 5: International Publication No. 2013 / 100014 Pamphlet

[0013] Non-patent literature

[0014] Non-patent document 1: Cancer Letters, 1999, Vol. 144, p177-182

[0015] Non-patent document 2: Oncology Reports, 2002, Vol. 9, p1319-1322

[0016] Non-patent document 3: YAKUGAKU ZASSHI, 2008, Vol. 128, p307-316

[0017] Non-patent document 4: Journal of Clinical Oncology, 2006, Vol. 24, p3946-3952

[0018] Non-patent document 5: Journal of Clinical Oncology, 2009, Vol. 23, p3778-3785 Summary of the invention

[0019] Technical issues to be solved by the invention

[0020] In recent years, antitumor drugs are not given alone, but are widely used in combination therapy. However, when any antitumor drugs are used in combination, it is not clear whether the antitumor effects of these drugs are enhanced or the effects cancel each other out.

[0021] The present invention aims to provide an antitumor agent, an antitumor kit, and an antitumor effect enhancer having superior antitumor effects compared to gemcitabine, an antitumor platinum complex (platinum preparation), and combination therapy thereof.

[0022] Means for solving technical problems

[0023] Therefore, the present inventors have studied the combined use of various drugs and found that a significant antitumor effect is exerted by the combined use of an antitumor platinum complex and Compound A, thereby completing the present invention.

[0024] That is, the present invention provides the following.

[0025] (1) An antitumor agent comprising an antitumor platinum complex and 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof.

[0026] (2) The antitumor agent according to (1), wherein

[0027] The antitumor platinum complex is at least one selected from the group consisting of cisplatin, oxaliplatin, and carboplatin.

[0028] (3) The antitumor agent according to (1) or (2), wherein

[0029] The above tumors are testicular tumors, bladder cancer, renal pelvis / ureter tumors, prostate cancer, testicular cancer, ovarian cancer, head and neck cancer, non-small cell lung cancer, small cell lung cancer, esophageal cancer, cervical cancer, glioma, neuroblastoma, gastric cancer, osteosarcoma, germ cell tumors (testicular tumors, ovarian tumors, extragonadal tumors), malignant pleural mesothelioma, biliary tract cancer, colon cancer, rectal cancer, small intestine cancer, malignant lymphoma, breast cancer, pancreatic cancer, liver cancer, kidney cancer, melanoma, leukemia or multiple myeloma.

[0030] (4) The antitumor agent according to any one of (1) to (3), wherein

[0031] The above tumor is bladder cancer, ovarian cancer, bile duct cancer, breast cancer or pancreatic cancer.

[0032] (5) The antitumor agent according to (4), wherein

[0033] The above-mentioned bile duct cancer is cholangiocarcinoma.

[0034] (6) An antitumor effect enhancer comprising 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof used in combination with an antitumor platinum complex.

[0035] (7) An antitumor kit comprising: a preparation containing an antitumor platinum complex; and a preparation containing 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof.

[0036] (8) An antitumor agent comprising 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof used in combination with an antitumor platinum complex.

[0037] (6-1) The anti-tumor effect enhancer according to (6), wherein

[0038] The antitumor platinum complex is at least one selected from the group consisting of cisplatin, oxaliplatin, and carboplatin.

[0039] (7-1) The antitumor kit according to (7), wherein

[0040] The antitumor platinum complex is at least one selected from the group consisting of cisplatin, oxaliplatin, and carboplatin.

[0041] (8-1) The antitumor agent according to (8), wherein

[0042] The antitumor platinum complex is at least one selected from the group consisting of cisplatin, oxaliplatin, and carboplatin.

[0043] (9) A method of using an antitumor platinum complex and 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof for treating tumors and for treating bladder cancer, ovarian cancer, biliary tract cancer, breast cancer or pancreatic cancer, the method comprising the step of administering a therapeutically effective amount to a subject (including a human mammal) in need of such treatment.

[0044] (10) A method for treating a tumor, characterized in that:

[0045] A therapeutically effective amount of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof for combined treatment and a therapeutically effective amount of an antitumor platinum complex for combined treatment are administered to a subject.

[0046] (11) A method for treating a tumor, characterized in that:

[0047] A therapeutically effective amount of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof for combined treatment and a therapeutically effective amount of an antitumor platinum complex for combined treatment are administered to a subject simultaneously, separately, continuously or at intervals.

[0048] (12) Use of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof for producing an antitumor agent in combination with an antitumor platinum complex.

[0049] (13) Use of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof for use in an antitumor agent in combination with an antitumor platinum complex.

[0050] (14) 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof, which is used for treating tumors by administering it together with an antitumor platinum complex as a single dosage form or as separate dosage forms.

[0051] (15) The antitumor agent according to (1) or (2), wherein

[0052] The daily dosage of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof is 20 to 200 mg / m 2 .

[0053] (16) The antitumor agent according to (15), wherein

[0054] The antitumor platinum complex is cisplatin, and the daily dosage is 1 to 100 mg / m 2 .

[0055] (17) The antitumor agent according to (15), wherein

[0056] The antitumor platinum complex is carboplatin, and the daily dosage is 50 to 1000 mg / m 2 .

[0057] (18) The antitumor agent according to (15), wherein

[0058] The antitumor platinum complex is oxaliplatin, and the daily dosage is 10 to 500 mg / m 2 .

[0059] Effects of the Invention

[0060] Compound A or its salt exerts a significant antitumor effect by being used in combination with an antitumor platinum complex. That is, compared with gemcitabine alone, antitumor platinum complex alone, or gemcitabine and an antitumor platinum complex in combination, the antitumor agent and antitumor kit of the present invention have excellent antitumor effects. The antitumor effect enhancer of the present invention can enhance the antitumor effect by being combined with an antitumor platinum complex for administration. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] Figure 1 This is a graph showing the effect of combined use of Compound A and cisplatin on the cell viability of SUIT-2 cells derived from human pancreatic cancer.

[0062] Figure 2 This is a graph showing the effect of combined use of Compound A and oxaliplatin on the cell viability of SUIT-2 cells derived from human pancreatic cancer.

[0063] Figure 3 This is a graph showing the effect of combined use of Compound A and cisplatin on the cell viability of human ovarian cancer cell line ES-2.

[0064] Figure 4 This is a graph showing the effect of combined use of Compound A and cisplatin on the cell viability of human ovarian cancer cell line SK-OV-3.

[0065] Figure 5 This is a graph showing the effect of combined use of Compound A and carboplatin on the cell viability of human ovarian cancer cell line SK-OV-3.

[0066] Figure 6 This is a graph showing the effect of combined use of Compound A and carboplatin on the cell viability of human ovarian cancer cell line ES-2.

[0067] Figure 7 This is a graph showing the effect of combined use of Compound A and cisplatin on the cell viability of HuCC-T1 cells, a cell line derived from human cholangiocarcinoma.

[0068] Figure 8 This is a graph showing the effect of combined use of Compound A and cisplatin on the cell viability of TFK-1 cells derived from human cholangiocarcinoma in spheroid culture.

[0069] Fig. 9 This is a graph showing the effect of combined use of Compound A and cisplatin on the cell viability of HuCC-T1, a human cholangiocarcinoma cell line, in spheroid culture.

[0070] Fig.10 This is a graph showing the effect of combined use of Compound A and cisplatin on the cell viability of HCC1954 cells derived from human breast cancer in spheroid culture. DETAILED DESCRIPTION

[0071] Furthermore, the range represented by "to" includes both end values ​​unless otherwise specified.

[0072] The "subject" refers to mammals such as humans, mice, monkeys, and livestock that need prevention or treatment, and is preferably a human that needs prevention or treatment.

[0073] "Prevention" refers to the obstruction of disease progression, reduction of the risk of disease progression, or delay of disease progression.

[0074] "Treatment" refers to improvement or suppression (maintenance or delay) of the progress of a target disease or condition.

[0075] "Treatment" refers to the prevention or treatment of various diseases.

[0076] "Tumor" refers to a benign tumor or a malignant tumor.

[0077] "Benign tumor" refers to a tumor whose cells and arrangement resemble the normal cells from which they originate and which is not invasive or metastatic.

[0078] The term "malignant tumor" refers to a tumor whose morphology or arrangement of tumor cells is different from the normal cells from which they originate and which shows invasiveness or metastasis.

[0079] Hereinafter, the present invention will be described in detail.

[0080] The antitumor agent of the present invention comprises an antitumor platinum complex and 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine (Compound A) or a salt thereof. Furthermore, the antitumor agent of the present invention is a combination of an antitumor platinum complex and Compound A or a salt thereof.

[0081] In the present invention, "combination" refers to a combination used for the combined use of compounds, including the following two forms, namely, the form of using each substance in combination when administering and the form of a mixture (compound). In the present invention, "combination" does not simply mean that the administration period of the compound of the present invention and the antitumor platinum complex is exactly the same. As long as the mode of supplying the compound of the present invention and the antitumor platinum complex is included in one administration schedule, the form of supplying these simultaneously or separately is "combination". In the case of separate administration, the antitumor platinum complex can be supplied after the compound of the present invention is supplied in advance. In addition, the compound of the present invention can also be supplied after the antitumor platinum complex is supplied in advance.

[0082] First, Compound A or a salt thereof will be described.

[0083] Examples of the salt include pharmaceutically acceptable salts, and specific examples include mineral acid salts, organic carboxylic acid salts, and sulfonates. Preferred salts include mineral acid salts and sulfonates.

[0084] As mineral acid salts, for example, hydrochlorides, hydrobromides, hydroiodides, nitrates, phosphates and sulfates can be mentioned, preferably hydrochlorides, hydroiodides, nitrates or sulfates, and more preferably hydrochlorides. As organic carboxylates, for example, formates, acetates, citrates, oxalates, fumarates, maleates, succinates, malates, tartrates, aspartates, trichloroacetates and trifluoroacetates can be mentioned. As sulfonates, for example, methanesulfonates, benzenesulfonates, p-toluenesulfonates, mesitylenesulfonates and naphthalenesulfonates can be mentioned, preferably methanesulfonates.

[0085] The salt of compound A can be an anhydride, a hydrate or a solvate. When simply referred to as a "salt" in this specification, its form can be an anhydride, a hydrate or a solvate. When referred to as "anhydride" in this specification, except for the case of special records, it refers to the case of being in a state of neither a hydrate nor a solvate. Even if it is a substance that does not originally form a hydrate or a solvate, the salt of compound A that does not have crystal water, hydration water and an interacting solvent is also included in the so-called "anhydride" in the present invention. Anhydride is sometimes also referred to as "anhydrous". When the salt of compound A is a hydrate, the number of hydration waters is not particularly limited, and it can be a monohydrate, a dihydrate, etc. As examples of solvates, methanolates, ethanolates, propanolates and 2-propanolates can be cited.

[0086] Specific examples of particularly preferred salts of Compound A are as follows.

[0087] 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine methanesulfonate;

[0088] 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine hydrochloride;

[0089] 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine 1 / 2 sulfate;

[0090] 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine nitrate; and

[0091] The hydroiodide salt of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine; and the anhydride of any of the above salts.

[0092] In the present invention, the compound A or a salt thereof may be used alone or in combination of two or more.

[0093] Next, the method for producing compound A or its salt is described. Compound A can be produced, for example, by the method described in Patent Document 1 and Journal of Organic Chemistry, 1999, Vol. 64, pp. 7912-7920. A salt of compound A or its hydrate or solvate can be produced, for example, by the method described in Patent Document 4.

[0094] The compound A or a salt thereof of the present invention can be used as an antitumor agent and can be used as an active ingredient of a pharmaceutical composition.

[0095] (Antitumor platinum complex)

[0096] In the present invention, examples of the antitumor platinum complex (platinum complex having an antitumor effect) include cisplatin, carboplatin, oxaliplatin, nedaplatin, zoniplatin, enloplatin, ormaplatin, roboplatin, siroplatin, lobaplatin, miplatin, or spiroplatin.

[0097] The antitumor platinum complex of the present invention is preferably cisplatin, carboplatin, oxaliplatin or nedaplatin, and more preferably cisplatin, carboplatin or oxaliplatin.

[0098] These antitumor platinum complexes can be produced by known methods.

[0099] Furthermore, these antitumor platinum complexes can also be obtained by purchasing commercial products. For example, cisplatin is commercially available as Randa (registered trademark) by Nippon Kayaku Co., Ltd., Platosin (registered trademark) by Pfizer Japan Inc., Cisplamerck (registered trademark) by Merck KgaA, and Briplatin (registered trademark) by Bristol-Myers Squibb Company. Carboplatin is commercially available as Paraplatin (registered trademark) by Bristol-Myers Squibb Company and Carbomerck (registered trademark) by Merck KgaA. Oxaliplatin is commercially available as Elplat (registered trademark) or Eloxatin (registered trademark) by Yakult Honsha Co., Ltd. Nedaplatin is commercially available as Aqupla (registered trademark) by Nichi-Iko Pharmaceutical Co., Ltd. Lobaplatin is commercially available from Chugai Pharmaceutical Co., Ltd. as Lobaplatin hydrochloride or Lobaplatin (registered trademark).

[0100] In the present invention, the antitumor platinum complex may also form a pharmaceutically acceptable salt with an acid or a base (hereinafter, sometimes referred to as "its salt"). The antitumor platinum complex of the present invention also includes these pharmaceutically acceptable salts. The antitumor platinum complex may be used alone or may contain two or more. As the antitumor platinum complex, in addition to the antitumor platinum complex or its salt, it may also be a composition containing these.

[0101] Examples of the salt include pharmaceutically acceptable salts, and specific examples include salts of generally known basic groups such as an amino group and acidic groups such as a hydroxyl group and a carboxyl group.

[0102] Examples of the salt in the basic group include salts with mineral acids such as hydrochloric acid, hydrobromic acid, nitric acid and sulfuric acid; salts with organic carboxylic acids such as formic acid, acetic acid, citric acid, oxalic acid, fumaric acid, maleic acid, succinic acid, malic acid, tartaric acid, aspartic acid, trichloroacetic acid and trifluoroacetic acid; and salts with sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, mesitylenesulfonic acid and naphthalenesulfonic acid.

[0103] Examples of the salt of the acidic group include salts with alkali metals such as sodium and potassium; salts with alkaline earth metals such as calcium and magnesium; ammonium salts; and salts with nitrogen-containing organic bases such as trimethylamine, triethylamine, tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, diethylamine, dicyclohexylamine, procaine, dibenzylamine, N-benzyl-β-phenylethylamine, 1-diphenylhydroxymethylamine, and N,N'-dibenzylethylenediamine.

[0104] Compound A is an antitumor agent having an excellent DNA synthesis inhibitory effect. When Compound A is used in combination with an antitumor platinum complex, it is expected to have an effect of enhancing the antitumor effect of the antitumor platinum complex without showing a significant deterioration in toxicity.

[0105] (Antitumor Agents)

[0106] According to the present invention, the following antitumor agent can be provided:

[0107] An antitumor agent comprising an antitumor platinum complex and 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof; and

[0108] An antitumor agent comprising 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof used in combination with an antitumor platinum complex.

[0109] The antitumor agent of the present invention may contain additives such as excipients, binders, lubricants, disintegrants, colorants, flavoring agents, emulsifiers, surfactants, solubilizers, suspending agents, isotonic agents, buffers, preservatives, antioxidants, stabilizers and absorption promoters that are commonly used for formulation.

[0110] The antitumor agent of the present invention comprising an antitumor platinum complex and compound A or a salt thereof may be a single-dose formulation comprising an antitumor platinum complex and compound A or a salt thereof, or may be a two-dose formulation comprising an antitumor platinum complex and compound A or a salt thereof. Preferably, the two-dose formulation comprises the antitumor platinum complex and compound A or a salt thereof as separate formulations.

[0111] When the antitumor platinum complex and compound A or its salt are used as separate preparations, each preparation can be administered to the subject simultaneously, separately, and continuously or at intervals. In addition, the administration method of the composition containing the antitumor platinum complex can be the same as the administration method of the composition containing compound A, or it can be different (for example, oral administration and injection).

[0112] Examples of the administration route of the antitumor agent of the present invention include intravenous, intraarterial, intrarectal, intraperitoneal, intramuscular, intratumoral or intravesical injection, oral administration, transdermal administration and / or suppository.

[0113] As the route of administration, it is preferably non-oral administration. For example, intravenous injection (intravenous injection), intramuscular injection, intraperitoneal injection, subcutaneous injection, intraocular injection and / or intramedullary injection such as drip can be enumerated. As the method of administration, the administration carried out by syringe or drip can be enumerated.

[0114] The dosage and administration method of compound A or its salt can be 1 to 2000 mg / m per day. 2 The dosage and method of administration are not limited to these.

[0115] The daily dosage of compound A or its salt is 20 mg / m 2 Above, preferably 40 mg / m 2 More preferably 60 mg / m 2 More preferably, 80 mg / m 2 The upper limit of the daily dosage is 200 mg / m 2 , preferably 150mg / m 2 , more preferably 120 mg / m 2 , especially preferably 100 mg / m 2 The daily dosage is more preferably 40 to 200 mg / m 2 , more preferably 40 to 150 mg / m 2 , more preferably 80 to 150 mg / m 2 , and more preferably 80 to 120 mg / m 2 .

[0116] In another embodiment, the daily dosage is preferably 20 to 120 mg / m 2 , more preferably 40 to 120 mg / m 2 , more preferably 40 to 100 mg / m 2 , more preferably 60 to 100 mg / m 2 .

[0117] By setting the dosage range as these, side effects can be minimized and the therapeutic effect as an antitumor agent can be maximized.

[0118] As a method of administering compound A or a salt thereof, the single dose may be set to 20 to 200 mg / m 2 , a step of administering once a week for 3 weeks and then stopping the administration in the 4th week. In this case, the single dose is the same as the above-mentioned daily dose, but is preferably 40 to 200 mg / m 2 , more preferably 40 to 150 mg / m 2 , more preferably 80 to 150 mg / m 2 In another embodiment, the single dose is preferably 20 to 120 mg / m 2 , more preferably 40 to 120 mg / m 2 , more preferably 40 to 100 mg / m 2 .

[0119] The antitumor platinum complex can be administered according to known clinical practice. The dosage and administration schedule can be changed according to the specific disease symptoms and the overall symptoms of the patient. For example, for adults, it can be administered orally or parenterally (for example, by injection, drip, and administration to the rectum), usually 1 to 2000 mg / m per day for adults. 2 The dosage is not particularly limited, but is, for example, 1 to 2000 mg / m 2 , preferably 10 to 2000 mg / m 2 , more preferably 50 to 1000 mg / m 2 , more preferably 100 to 500 mg / m 2 , the dosage can be divided into 1 to 3 times per day for administration. In the case where the patient experiences excessive toxicity, the dosage needs to be reduced. In addition to the combined therapy of the present invention, when one or more additional chemotherapeutic agents are used, the dosage and dosing schedule can also be changed. The dosing schedule can be determined by the doctor treating the specific patient.

[0120] The dosage or blending amount of the antitumor platinum complex and compound A or a salt thereof contained in the antitumor agent of the present invention is not particularly limited as long as it is within the range of the enhancement effect of the antitumor effect.

[0121] The amount of the compound A of the present invention used varies depending on each combination with the antitumor platinum complex, but is, for example, about 0.0001 to 10000 times (weight ratio), preferably about 0.001 to 1000 times (weight ratio) of the antitumor platinum complex.

[0122] More specifically, when the compound A and cisplatin are combined, the dosage of the compound A of the present invention may be set to 20 to 200 mg / m per day for adults, although it is not particularly limited. 2 , preferably 40 to 200 mg / m 2 , more preferably 40 to 150 mg / m 2 , more preferably 80 to 150 mg / m 2 The dosage of cisplatin is set at 1-2000 mg / m2 per day for adults. 2 , preferably 10 to 2000 mg / m 2 , more preferably 50 to 1000 mg / m 2 , more preferably 100 to 500 mg / m 2 Furthermore, the dosage of the compound of the present invention is set to about 0.0001 to 10000 times (weight ratio) that of cisplatin, preferably about 0.001 to 1000 times (weight ratio).

[0123] In another embodiment, for example, the dosage of cisplatin may be set to 1 to 100 mg / m2 per day for adults. 2 , preferably 5 to 50 mg / m 2 , more preferably 10 to 50 mg / m 2 , more preferably 10 to 30 mg / m 2 Furthermore, the dosage of the compound of the present invention is set to about 0.2 to 200 times (weight ratio) that of cisplatin, preferably about 1.6 to 15 times (weight ratio).

[0124] Furthermore, when the compound of the present invention is combined with carboplatin, there is no particular limitation, but for example, the dosage of the compound A of the present invention may be set to 20 to 200 mg / m per day for adults. 2 , preferably 40 to 200 mg / m 2 , more preferably 40 to 150 mg / m 2 , more preferably 80 to 150 mg / m 2 The dosage of carboplatin is set at 10 to 2000 mg / m per day for adults. 2 , preferably 50 to 1000 mg / m 2 , more preferably 100 to 500 mg / m 2 Furthermore, the dosage of the compound of the present invention is set to about 0.0001 to 10000 times (weight ratio) of carboplatin, preferably about 0.001 to 1000 times (weight ratio), more preferably about 0.01 to 20 times (weight ratio), and further preferably about 0.16 to 1.5 times (weight ratio).

[0125] Furthermore, when the compound of the present invention is combined with oxaliplatin, the dosage is not particularly limited, but for example, the dosage of the compound A of the present invention may be set to 20 to 200 mg / m per day for adults. 2 , preferably 40 to 200 mg / m 2 , more preferably 40 to 150 mg / m 2 , more preferably 80 to 150 mg / m 2 The dosage of oxaliplatin is set at 10 to 2000 mg / m per day for adults. 2 , preferably 50 to 1000 mg / m 2 , more preferably 100 to 500 mg / m 2 Furthermore, the dosage of the compound of the present invention is set to about 0.0001 to 10000 times (weight ratio) that of oxaliplatin, preferably about 0.001 to 1000 times (weight ratio).

[0126] In another embodiment, the dosage of oxaliplatin may be set to 10 to 500 mg / m2 per day for adults. 2 , preferably 50 to 500 mg / m 2 , more preferably 50 to 150 mg / m 2 Furthermore, the dosage of the compound of the present invention is set to about 0.04 to 20 times (weight ratio) that of oxaliplatin, preferably about 0.53 to 3 times (weight ratio).

[0127] Examples of dosage forms of the antitumor agent of the present invention include tablets, capsules, powders, syrups, granules, pills, suspensions, emulsions, liquids, suppositories, eye drops, nose drops, ear drops, patches, ointments and injections, but injections are preferred. These dosage forms can be manufactured by conventional preparation methods known to those skilled in the art.

[0128] The antitumor agent of the present invention is preferably an anti-malignant tumor agent and can be used as an anticancer agent.

[0129] The antitumor agent of the present invention can be effectively used for the treatment of various types of tumors including, for example, testicular tumors, bladder cancer, renal pelvis / ureter tumors, prostate cancer, testicular cancer, ovarian cancer, head and neck cancer, non-small cell lung cancer, small cell lung cancer, esophageal cancer, cervical cancer, glioma, neuroblastoma, gastric cancer, osteosarcoma, germ cell tumors (testicular tumors, ovarian tumors, extragonadal tumors), malignant pleural mesothelioma, biliary tract cancer, colon cancer, rectal cancer, small intestine cancer, malignant lymphoma, breast cancer, pancreatic cancer, liver cancer, kidney cancer, melanoma, leukemia, multiple myeloma, or tumors of other organs. Among them, preferred are testicular tumors, bladder cancer, renal pelvis / ureter tumors, prostate cancer, ovarian cancer, head and neck cancer, non-small cell lung cancer, small cell lung cancer, esophageal cancer, cervical cancer, neuroblastoma, gastric cancer, osteosarcoma, germ cell tumors (testicular tumors, ovarian tumors, extragonadal tumors), malignant pleural mesothelioma, biliary tract cancer, colon cancer, rectal cancer, malignant lymphoma, breast cancer, and pancreatic cancer; more preferred are bladder cancer, ovarian cancer, biliary tract cancer, breast cancer, or pancreatic cancer; further preferred are biliary tract cancer, and it is especially effective for the treatment of cholangiocarcinoma.

[0130] (Anti-tumor effect enhancer)

[0131] Furthermore, the present invention relates to an antitumor effect enhancer comprising compound A or a salt thereof for enhancing the antitumor effect of an antitumor platinum complex in cancer patients.

[0132] That is, according to the present invention, there is provided an antitumor effect enhancer comprising 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof used in combination with an antitumor platinum complex.

[0133] The anti-tumor effect enhancer of the present invention may contain additives such as excipients, binders, lubricants, disintegrants, colorants, flavoring agents, emulsifiers, surfactants, solubilizers, suspending agents, isotonic agents, buffers, preservatives, antioxidants, stabilizers and absorption promoters that are commonly used for formulation.

[0134] The antitumor effect enhancer of the present invention can be administered to a subject simultaneously with the antitumor platinum complex, separately, continuously or at intervals.

[0135] As the administration route of the anti-tumor effect enhancer of the present invention, parenteral administration is preferred. For example, intravenous injection (intravenous injection) such as drip, intramuscular injection, intraperitoneal injection, subcutaneous injection, intraocular injection and intramedullary injection can be mentioned. As the administration method, administration by syringe or drip can be mentioned.

[0136] The dosage or amount of Compound A or a salt thereof contained in the antitumor effect enhancer of the present invention is not particularly limited as long as it is within the range in which the antitumor effect enhancing effect is exerted.

[0137] The amount of the compound A of the present invention used varies depending on each combination with the antitumor platinum complex, but is, for example, about 0.0001 to 10000 times (weight ratio), preferably about 0.001 to 1000 times (weight ratio) of the antitumor platinum complex.

[0138] The dosage and administration method of the compound A or a salt thereof contained in the antitumor effect enhancer of the present invention can be 1 to 2000 mg / m per day. 2 The dosage and method of administration are not limited to these.

[0139] The daily dosage of compound A or its salt is 20 mg / m 2 Above, preferably 40 mg / m 2 More preferably 60 mg / m 2 More preferably, 80 mg / m 2 The upper limit of the daily dosage is 200 mg / m 2 , preferably 150mg / m 2 , more preferably 120 mg / m 2 , especially preferably 100 mg / m 2 The daily dosage is more preferably 40 to 200 mg / m 2 , more preferably 40 to 150 mg / m 2 , more preferably 80 to 150 mg / m 2 , more preferably 80 to 120 mg / m 2 .

[0140] In another embodiment, the daily dosage is preferably 20 to 120 mg / m 2 , more preferably 40 to 120 mg / m 2 , more preferably 40 to 100 mg / m 2 , more preferably 60 to 100 mg / m 2 .

[0141] By setting the dosage range as these, side effects can be minimized and the therapeutic effect as an antitumor agent can be maximized.

[0142] As a method of administering compound A or a salt thereof, the single dose may be set to 20 to 200 mg / m 2, a step of administering once a week for 3 weeks and then stopping the administration in the 4th week. In this case, the single dose is the same as the above-mentioned daily dose, but is preferably 40 to 200 mg / m 2 , more preferably 40 to 150 mg / m 2 , more preferably 80 to 150 mg / m 2 In another embodiment, the single dose is preferably 20 to 120 mg / m 2 , more preferably 40 to 120 mg / m 2 , more preferably 40 to 100 mg / m 2 .

[0143] The antitumor effect enhancer of the present invention is preferably an anti-malignant tumor agent effect enhancer, and can be used as an anticancer effect enhancer.

[0144] The anti-tumor effect enhancer of the present invention can be effectively used for the treatment of various types of tumors including testicular tumors, bladder cancer, renal pelvis / ureter tumors, prostate cancer, testicular cancer, ovarian cancer, head and neck cancer, non-small cell lung cancer, small cell lung cancer, esophageal cancer, cervical cancer, glioma, neuroblastoma, gastric cancer, osteosarcoma, germ cell tumors (testicular tumors, ovarian tumors, extragonadal tumors), malignant pleural mesothelioma, biliary tract cancer, colon cancer, rectal cancer, small intestine cancer, malignant lymphoma, breast cancer, pancreatic cancer, liver cancer, kidney cancer, melanoma, leukemia, multiple myeloma or tumors of other organs. Among them, preferred are testicular tumors, bladder cancer, renal pelvis / ureter tumors, prostate cancer, ovarian cancer, head and neck cancer, non-small cell lung cancer, small cell lung cancer, esophageal cancer, cervical cancer, neuroblastoma, gastric cancer, osteosarcoma, germ cell tumors (testicular tumors, ovarian tumors, extragonadal tumors), malignant pleural mesothelioma, biliary tract cancer, colon cancer, rectal cancer, malignant lymphoma, breast cancer, and pancreatic cancer; more preferred are bladder cancer, ovarian cancer, biliary tract cancer, breast cancer, or pancreatic cancer; further preferred are biliary tract cancer, and it is especially effective for the treatment of cholangiocarcinoma.

[0145] (Anti-tumor kit)

[0146] According to the present invention, there is provided an antitumor kit comprising a preparation containing an antitumor platinum complex and a preparation containing 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof.

[0147] The antitumor kit of the present invention is a kit comprising a combination of (a) an antitumor platinum complex and (b) compound A or a salt thereof.

[0148] Furthermore, in the above kit, (a) the antitumor platinum complex and (b) compound A or a salt thereof can be provided in various known formulation forms, and can be accommodated in various commonly used containers according to the formulation forms.

[0149] Furthermore, in the above-mentioned kit, (a) the antitumor platinum complex and (b) compound A or its salt may be contained in different containers, or may be mixed and contained in the same container. Preferably, (a) the antitumor platinum complex and (b) compound A or its salt are contained in different containers.

[0150] The present invention provides a method for treating tumors, preferably bladder cancer, ovarian cancer, biliary cancer, breast cancer or pancreatic cancer, using an antitumor platinum complex and 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof, comprising the step of administering a therapeutically effective amount to a subject (including a human mammal) in need of such treatment.

[0151] Furthermore, the present invention provides a method for treating tumors, characterized in that a therapeutically effective amount of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof for combined treatment and a therapeutically effective amount of an antitumor platinum complex for combined treatment are administered to a subject.

[0152] Furthermore, the present invention provides a method for treating tumors, characterized in that a therapeutically effective amount of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof for combined treatment and a therapeutically effective amount of an antitumor platinum complex for combined treatment are administered to a subject simultaneously, separately, continuously or at intervals.

[0153] An antitumor agent which can be used for producing 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof, and which is combined with an antitumor platinum complex.

[0154] Use of an antitumor agent which can be used for 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof in combination with an antitumor platinum complex.

[0155] According to the present invention, 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof can be obtained for treating tumors by administering it together with an antitumor platinum complex as a single dosage form or as separate dosage forms.

[0156] [Example]

[0157] The present invention will be described in further detail below with reference to Examples and Test Examples, but the present invention is not limited to these Examples and the like.

[0158] (Example 1)

[0159] The methanesulfonate of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine (Compound A) was synthesized by the method described in International Publication No. 2013 / 146833.

[0160] (Test Example 1)

[0161] Evaluation of the antitumor activity of cisplatin and oxaliplatin combined with oxaliplatin against the human pancreatic cancer cell line SUIT-2

[0162] As test substances, gemcitabine (hereinafter also referred to as Gemcitabine or Gem), cisplatin (hereinafter also referred to as Cisplatin), oxaliplatin (hereinafter also referred to as Oxaliplatin), and mesylate of Compound A were used.

[0163] Gemcitabine was prepared by dissolving gemcitabine hydrochloride (manufactured by Plantex) in dimethyl sulfoxide (DMSO). Cisplatin (manufactured by Wako Pure Chemical Industries, Ltd., Cat. #039-20091) and oxaliplatin (manufactured by Tokyo Chemical Industry Co., Ltd., Cat. #00372) were dissolved in serum-free medium RPMI-1640 (manufactured by Thermo Fisher Scientific Inc., Cat. #11875-119).

[0164] SUIT-2 cells, a human pancreatic cancer cell line, were subcultured using RPMI-1640 medium containing 10% serum (Thermo Fisher Scientific Inc., Cat. #10437-028). In this experiment, all cells were cultured under CO 2 Incubator (37°C, 5% CO 2 , water vapor saturation). Diluted to 15000 cells / well / 100μL with 10% serum medium and seeded in 96-well plates. On the second day, the culture supernatant of each well was removed, washed twice with 150μL of serum-free medium, and cultured with 100μL of serum-free medium for 3 days.

[0165] The mesylate of compound A and Gemcitabine were dissolved in DMSO to prepare 100 mmol / L DMSO solutions. The mixture was diluted with DMSO in sequence to prepare DMSO solutions with a concentration of 1000 times the final treatment concentration. Cisplatin and Oxaliplatin were dissolved in serum-free culture medium to prepare 30 μmol / L Cisplatin solution and 60 μmol / L Oxaliplatin solution. The DMSO diluted solutions of compound A and Gemcitabine were diluted with Cisplatin solution (30 μmol / L) or Oxaliplatin solution (60 μmol / L) to prepare treatment solutions with a concentration of 6 times the final treatment concentration. For compound A and Gemcitabine, the maximum concentration was set to 10 μmol / L, and 9 concentrations were combined with Cisplatin or Oxaliplatin at a common ratio of 1 / 3 for use.

[0166] 20 μL of compound A or Gemcitabine diluted with Cisplatin solution (30 μmol / L) or Oxaliplatin solution (60 μmol / L) was added to each well. In addition, a group in which only a solvent without a drug was added to the wells seeded with cells (positive control group) and a group in which only a solvent without a drug was added to the wells in which only a culture medium was added (negative control group) were set up. Both were set to n = 3 wells.

[0167] After adding the drug, the cells were cultured for 3 days, and the cell survival rate was evaluated using CellTiter Glo (registered trademark) Reagent (Cat. #G7570 manufactured by PROMEGA Co.) using the intracellular ATP level as an indicator. The concentration that inhibited the cell survival rate by 50% (IC50) was calculated using XLFit software Ver.3 (registered trademark) (manufactured by CTC).

[0168] The cell survival rate of each well was determined by setting the luminescence signal of the negative control group to 0% and the luminescence signal of the positive control group to 100%. The mean value and standard deviation of the cell survival rate of each treatment group were calculated to prepare Tables 1 and 2.

[0169] Results of combined use test with Cisplatin 5μmol / L

[0170] [Table 1]

[0171] The average cell survival rate

[0172] Concentration (nM) 10000.0 3333.3 1111.1 370.4 123.5 41.2 13.7 4.6 1.5 Gemcitabine 22 32 54 73 82 94 99 97 98 Compound A 30 27 26 28 28 33 47 80 96

[0173] Standard deviation of cell viability

[0174] Concentration (nM) 10000.0 3333.3 1111.1 370.4 123.5 41.2 13.7 4.6 1.5 Gemcitabine 1.7 1.5 2.3 1.3 1.6 2.9 6.0 2.6 2.5 Compound A 15.1 10.1 4.8 5.3 1.3 7.5 4.5 0.5 2.8

[0175] Results of combined use test with Oxaliplatin 10μmol / L

[0176] [Table 2]

[0177] The average cell survival rate

[0178] Concentration (nM) 10000 3000 1000 300 100 30 10 3 1 Gemcitabine 61 80 89 96 101 97 97 102 98 Compound A 38 39 38 41 46 60 87 93 94

[0179] Standard deviation of cell viability

[0180] Concentration (nM) 10000 3000 1000 300 100 30 10 3 1 Gemcitabine 2.8 3.1 1.0 2.0 5.1 1.7 0.9 3.7 0.8 Compound A 2.2 2.7 2.9 4.7 2.6 1.2 4.1 2.5 7.2

[0181] Furthermore, the graphs made according to Table 1 and Table 2 are shown in Figure 1 and Figure 2 middle.

[0182] The IC50 value calculated according to Table 1 when gemcitabine and cisplatin are used together is 1138.3nmol / L (nM), and the IC50 value when compound A and cisplatin are used together is 12.9nmol / L. In addition, the IC50 value calculated according to Table 2 when gemcitabine and oxaliplatin are used together is more than 10,000nmol / L, and the IC50 value when compound A and oxaliplatin are used together is 73.4nmol / L. As described above, compound A significantly enhances the anti-tumor effect of cisplatin and oxaliplatin. It is believed that its effect is greater than gemcitabine as an existing drug. A more detailed description will be described later.

[0183] The combined effect of the present invention is evaluated using a combination index (CI) which is a quantitative index of the combined effect. CI can be calculated by the following formula according to Cancer Research, 2010, Vol. 70, pp. 440-446.

[0184] That is, if the drugs used in combination are drug 1 and drug 2, the CI at a certain drug concentration is

[0185] CI = {(cell survival rate when drug 1 and drug 2 are used together) ÷ 100} / {[(cell survival rate of drug 1) ÷ 100] × [(cell survival rate of drug 2) ÷ 100]}

[0186] CI=1:Additive effect

[0187] CI>1: antagonistic effect

[0188] CI<1:synergistic effect

[0189] The CI when cisplatin and gemcitabine were used together was 0.96, and the CI when cisplatin and compound A were used together was 0.36. The CI when oxaliplatin and gemcitabine were used together was 0.97, and the CI when oxaliplatin and compound A were used together was 0.44. CI <1, so the synergistic effect based on the combined use of compound A and cisplatin or oxaliplatin was confirmed. In addition, it is speculated that the smaller the CI value, the higher the synergistic effect, so it can be said that the synergistic effect of compound A is more obvious than gemcitabine, which is an existing drug.

[0190] (Test Example 2)

[0191] Evaluation of the antitumor activity of cisplatin combined with carboplatin against human ovarian cancer cell lines ES-2 and SK-OV-3

[0192] Furthermore, by the same method as in Test Example 1, the antitumor activity of cisplatin and carboplatin in combination against human ovarian cancer cell lines ES-2 and SK-OV-3 was evaluated.

[0193] As test substances, gemcitabine, cisplatin, carboplatin, and mesylate of Compound A were used.

[0194] Gemcitabine was prepared by dissolving gemcitabine hydrochloride (manufactured by Plantex) in DMSO. Cisplatin (manufactured by Wako Pure Chemical Industries, Ltd., Cat. #039-20091) and carboplatin (manufactured by Tokyo Chemical Industry Co., Ltd., Cat. #C2043) were dissolved in 1% serum (manufactured by Thermo Fisher Scientific Inc., Cat. #10437-028) and McCoy's 5a (manufactured by Thermo Fisher Scientific Inc., Cat. #16600-082).

[0195] ES-2 and SK-OV-3 cells, human ovarian cancer cell lines, were subcultured in 10% serum medium. In this experiment, all cells were cultured in CO 2 Incubator (37°C, 5% CO 2ES-2 cells were diluted to 10,000 cells / well / 100 μL using 10% serum medium, and SK-OV-3 cells were diluted to 15,000 cells / well / 100 μL, and seeded in 96-well plates. On the second day, the culture supernatant of each well was removed, and the wells were washed twice with 150 μL of 1% serum medium, and 100 μL of 1% serum medium was added and cultured for 3 days.

[0196] The mesylate of compound A and Gemcitabine were dissolved in DMSO to prepare 100 mmol / L DMSO solutions. The solutions were diluted with DMSO in sequence to prepare DMSO solutions with a concentration of 1000 times the final treatment concentration. Cisplatin and Carboplatin were dissolved in 1% serum culture medium to prepare 60 μmol / L Cisplatin solution and 600 μmol / L Carboplatin solution, respectively. The DMSO diluted solutions of compound A and Gemcitabine were diluted with Cisplatin solution (60 μmol / L) or Carboplatin solution (600 μmol / L) to prepare treatment solutions with a concentration of 6 times the final treatment concentration, respectively. For compound A and Gemcitabine, the maximum concentration was set to 10 μmol / L, and 9 concentrations were used in combination with Cisplatin or Carboplatin at a common ratio of 1 / 3.

[0197] 20 μL of compound A or Gemcitabine diluted with Cisplatin solution (60 μmol / L) or Carboplatin solution (600 μmol / L) was added to each well. In addition, a group in which only solvent without drug was added to the wells seeded with cells (positive control group) and a group in which only solvent without drug was added to the wells added with culture medium (negative control group) were set up. Both were set to n = 3 wells.

[0198] After adding the drug, the cells were cultured for 3 days, and the cell viability was evaluated using CellTiter Glo (registered trademark) Reagent (Cat. #G7570 manufactured by PROMEGA Co.) using the intracellular ATP level as an indicator. The concentration that inhibited the cell viability by 50% (IC50 value) was calculated using XLFit software Ver.3 (registered trademark) (manufactured by CTC).

[0199] The cell survival rate of each well was determined by setting the luminescence signal amount of the negative control group as 0% and the luminescence signal amount of the positive control group as 100%. The mean value and standard deviation of the cell survival rate of each treatment group were calculated to prepare Tables 3 to 6.

[0200] Results of combined use test with Cisplatin 10μmol / L

[0201] ES-2 cells

[0202] [Table 3]

[0203] The average cell survival rate

[0204] Concentration (nM) 10000 3000 1000 300 100 30 10 3 1 Gemcitabine 50 65 74 79 79 79 78 78 74 Compound A 21 23 23 26 37 68 77 78 78

[0205] Standard deviation of cell viability

[0206] Concentration (nM) 10000 3000 1000 300 100 30 10 3 1 Gemcitabine 3.4 3.7 2.9 3.1 2.5 4.7 4.7 5.5 1.4 Compound A 0.9 0.7 0.7 0.5 1.3 2.3 0.9 3.2 2.2

[0207] Results of combined use test with Cisplatin 10μmol / L

[0208] SK-OV-3 cells

[0209] [Table 4]

[0210] The average cell survival rate

[0211] Concentration (nM) 10000 3000 1000 300 100 30 10 3 1 Gemcitabine 30 59 79 83 83 83 84 84 82 Compound A 10 10 11 14 43 76 81 82 83

[0212] Standard deviation of cell viability

[0213] Concentration (nM) 10000 3000 1000 300 100 30 10 3 1 Gemcitabine 1.4 1.1 0.6 0.6 2.6 2.1 3.8 1.9 1.2 Compound A 0.5 0.3 0.4 0.9 1.6 3.3 3.0 1.9 3.2

[0214] Results of combined use test with Carboplatin 30μmol / L

[0215] SK-OV-3 cells

[0216] [Table 5]

[0217] The average cell survival rate

[0218] Concentration (nM) 10000 3000 1000 300 100 30 10 3 1 Gemcitabine 76 80 81 82 83 86 88 90 90 Compound A 17 20 41 73 80 82 84 88 90

[0219] Standard deviation of cell viability

[0220] Concentration (nM) 10000 3000 1000 300 100 30 10 3 1 Gemcitabine 5.6 4.9 1.2 0.9 0.7 0.5 1.4 2.9 1.4 Compound A 0.5 0.6 2.1 1.3 1.5 0.6 1.5 1.6 3.1

[0221] Results of combined use test with Carboplatin 100μmol / L

[0222] ES-2 cells

[0223] [Table 6]

[0224] The average cell survival rate

[0225] Concentration (nM) 10000 3000 1000 300 100 30 10 3 1 Gemcitabine 48 62 69 71 72 71 71 74 76 Compound A 21 22 24 28 36 57 69 70 73

[0226] Standard deviation of cell viability

[0227] Concentration (nM) 10000 3000 1000 300 100 30 10 3 1 Gemcitabine 1.5 1.8 1.7 0.7 1.3 1.6 1.3 2.5 2.4 Compound A 1.1 0.5 0.6 0.2 1.4 3.3 1.5 2.2 1.9

[0228] Furthermore, the graphs prepared according to Tables 3 to 6 are shown in Figures 3 to 6 middle.

[0229] In ES-2 cells, the IC50 value calculated according to Table 3 when gemcitabine and cisplatin were used in combination could not be calculated, and the IC50 value when compound A and cisplatin were used in combination was 63.2 nmol / L. In addition, in SK-OV-3 cells, the IC50 value calculated according to Table 4 when gemcitabine and cisplatin were used in combination was 3797.2 nmol / L, and the IC50 value when compound A and cisplatin were used in combination was 82.2 nmol / L.

[0230] In SK-OV-3 cells, the IC50 value calculated according to Table 5 when gemcitabine and carboplatin were used in combination could not be calculated, and the IC50 value when compound A and carboplatin were used in combination was 676.5 nmol / L. In addition, in ES-2 cells, the IC50 value calculated according to Table 6 when gemcitabine and carboplatin were used in combination was 10,000 nmol / L or more, and the IC50 value when compound A and carboplatin were used in combination was 46.0 nmol / L.

[0231] CI was calculated from Tables 3 to 6, and summarized to create Table 7.

[0232] [Table 7]

[0233]

[0234] CI<1, so a synergistic effect was confirmed by the combined use of Compound A and cisplatin or carboplatin. In addition, it is presumed that the smaller the CI value, the higher the synergistic effect, so it can be said that the synergistic effect of Compound A is more significant than that of gemcitabine, which is an existing drug.

[0235] In each evaluation, a synergistic effect based on the combined use with cisplatin or carboplatin was confirmed for Compound A, and the results obtained indicated that the synergistic effect of Compound A was more significant than that of gemcitabine, which is an existing drug.

[0236] (Test Example 3)

[0237] Evaluation of the antitumor activity of cisplatin combined with cisplatin against the human cholangiocarcinoma cell line HuCC-T1

[0238] Furthermore, by the same method as in Test Example 1, the antitumor activity of the compound in combination with cisplatin was evaluated against the human cholangiocarcinoma-derived cell line HuCC-T1.

[0239] As test substances, gemcitabine, cisplatin, and mesylate of compound A were used.

[0240] Gemcitabine was prepared by dissolving gemcitabine hydrochloride (manufactured by Plantex) in DMSO. Cisplatin (manufactured by Wako Pure Chemical Industries, Ltd., Cat. #039-20091) was dissolved in serum-free medium RPMI-1640 (manufactured by Thermo Fisher Scientific Inc., Cat. #11875-119).

[0241] HuCC-T1 cells, a human cholangiocarcinoma cell line, were subcultured using RPMI-1640 medium containing 10% serum (Thermo Fisher Scientific Inc., Cat. #10437-028). 2 Incubator (37°C, 5% CO 2 , water vapor saturation). Diluted to 15000 cells / well / 100μL with 10% serum medium and seeded in 96-well plates. On the second day, the culture supernatant of each well was removed, washed twice with 150μL of serum-free medium, and cultured with 100μL of serum-free medium for 3 days.

[0242] The mesylate of compound A and Gemcitabine were dissolved in DMSO to prepare 100 mmol / L DMSO solutions. The mixture was diluted with DMSO in sequence to prepare DMSO solutions with a concentration of 1000 times the final treatment concentration. Cisplatin was dissolved in serum-free medium to prepare a 60 μmol / L Cisplatin solution. The DMSO diluted solutions of compound A and Gemcitabine were diluted with Cisplatin solution (60 μmol / L) to prepare treatment solutions with a concentration of 6 times the final treatment concentration. For compound A and Gemcitabine, the maximum concentration was set to 10 μmol / L, and 9 concentrations were combined with Cisplatin at a common ratio of 1 / 3 for use.

[0243] 20 μL of compound A or Gemcitabine diluted with Cisplatin solution (60 μmol / L) was added to each well. In addition, a group in which only solvent without drug was added to the wells seeded with cells (positive control group) and a group in which only solvent without drug was added to the wells added with culture medium (negative control group) were set up. Both were set to n = 3 wells.

[0244] After adding the drug, the cells were cultured for 3 days, and the cell viability was evaluated using CellTiter Glo (registered trademark) Reagent (Cat. #G7570 manufactured by PROMEGA Co.) using the intracellular ATP level as an indicator. The concentration that inhibited the cell viability by 50% (IC50 value) was calculated using XLFit software Ver.3 (registered trademark) (manufactured by CTC).

[0245] The cell survival rate of each well was determined by setting the luminescence signal amount of the negative control group to 0% cell survival rate and the luminescence signal amount of the positive control group to 100%. The mean value and standard deviation of the cell survival rate of each treatment group were calculated to prepare Table 8.

[0246] Results of combined use test with Cisplatin 10μmol / L

[0247] [Table 8]

[0248] The average cell survival rate

[0249] Concentration (nM) 10000.0 3333.3 1111.1 370.4 123.5 41.2 13.7 4.6 1.5 Gemcitabine 56 83 103 113 116 113 116 113 114 Compound A 34 36 40 44 66 96 110 114 117

[0250] Standard deviation of cell viability

[0251] Concentration (nM) 10000.0 3333.3 1111.1 370.4 123.5 41.2 13.7 4.6 1.5 Gemcitabine 4.3 3.9 6.0 1.7 1.1 4.0 1.6 1.1 0.3 Compound A 4.1 2.2 1.2 8.0 12.6 5.0 2.3 2.8 0.3

[0252] Furthermore, the graph produced according to the above Table 8 is shown in Figure 7 middle.

[0253] The IC50 value calculated from Table 8 when gemcitabine and cisplatin were used together was 10,000 nmol / L or more, and the IC50 value when compound A and cisplatin were used together was 261.0 nmol / L. As described above, compound A significantly enhanced the antitumor effect of cisplatin. It is believed that the effect is greater than gemcitabine, which is an existing drug.

[0254] The CI when cisplatin and gemcitabine were used together was 0.90, and the CI when cisplatin and compound A were used together was 0.34. CI < 1, so a synergistic effect based on the combined use of compound A and cisplatin was confirmed. In addition, it is estimated that the smaller the CI value, the higher the synergistic effect, so it can be said that the synergistic effect of compound A is more significant than gemcitabine, which is an existing drug.

[0255] (Test Example 4)

[0256] Evaluation of the antitumor activity of cisplatin-based combination therapy against TFK-1 in cholangiocarcinoma cell line in spheroid culture

[0257] As test substances, gemcitabine, cisplatin, and mesylate of compound A were used.

[0258] Gemcitabine was prepared by dissolving gemcitabine hydrochloride (manufactured by Plantex) in dimethyl sulfoxide (DMSO). Cisplatin (manufactured by Wako Pure Chemical Industries, Ltd., Cat. #039-20091) was dissolved in DMSO.

[0259] TFK-1 cells, a human cholangiocarcinoma cell line, were subcultured using RPMI-1640 medium containing 10% serum (Thermo Fisher Scientific Inc., Cat. #10437-028). In this experiment, all cells were cultured under CO 2 Incubator (37°C, 5% CO 2 The cells were diluted to 5000 cells / well / 100 μL using 10% serum medium and seeded in an Ultra-Low adhesion 96-well plate (Cat. #7007 manufactured by Corning Incorporated). Spheroids were formed after 4 days of culture.

[0260] The mesylate of compound A and Gemcitabine were dissolved in DMSO to prepare 10mmol / L DMSO solutions. The solutions were diluted with DMSO in sequence to prepare DMSO solutions with a concentration of 1000 times the final treatment concentration. Cisplatin was dissolved in DMSO to prepare a 100mmol / L DMSO solution. The solutions were diluted with DMSO to prepare a DMSO solution with a concentration of 5000 times the final treatment concentration. Furthermore, the DMSO solution of Cisplatin was diluted with 10% serum medium to prepare a medium containing Cisplatin with a concentration of 10 times the final treatment concentration. The DMSO diluted solutions of compound A and Gemcitabine were diluted with the medium containing Cisplatin to prepare treatment solutions with a concentration of 10 times the final treatment concentration. For compound A and Gemcitabine, the maximum concentration was set to 10μmol / L, and 9 concentrations were combined with Cisplatin (10μmol / L) at a common ratio of 1 / 3 and used.

[0261] 10 μL of compound A or Gemcitabine diluted with cisplatin-containing culture medium was added to each well. In addition, a group in which only solvent without drug was added to the wells seeded with cells (positive control group) and a group in which only solvent without drug was added to the wells added with culture medium (negative control group) were set up. Both were set to n = 3 wells.

[0262] After adding the drug, the cells were cultured for 3 days, and the cell viability was evaluated using CellTiter Glo (registered trademark) Reagent (Cat. #G7570 manufactured by PROMEGA Co.) using the intracellular ATP level as an indicator. The concentration that inhibited the cell viability by 50% (IC50 value) was calculated using XLFit software Ver.3 (registered trademark) (manufactured by CTC).

[0263] The cell survival rate of each well was determined by setting the luminescence signal amount of the negative control group to 0% cell survival rate and the luminescence signal amount of the positive control group to 100%. The mean value and standard deviation of the cell survival rate of each treatment group were calculated to prepare Table 9.

[0264] Results of combined use test with Cisplatin 10μmol / L

[0265] [Table 9]

[0266] The average cell survival rate

[0267] Concentration (nM) 10000.0 3000.0 1000.0 300.0 100.0 30.0 10.0 3.0 1.0 Gemcitabine 66 77 83 83 90 89 97 97 97 Compound A 9 14 23 65 90 91 96 101 97

[0268] Standard deviation of cell viability

[0269] Concentration (nM) 10000.0 3000.0 1000.0 300.0 100.0 30.0 10.0 3.0 1.0 Gemcitabine 0.8 1.3 6.0 7.7 3.4 5.7 3.3 6.1 8.8 Compound A 0.3 0.3 1.5 2.2 3.9 3.9 6.2 6.0 0.5

[0270] Furthermore, the graph produced according to the above Table 9 is shown in Figure 8 middle.

[0271] The IC50 value when gemcitabine and cisplatin (10 μmol / L) were used together in spheroid culture was 10000 nmol / L or more, and the IC50 value when compound A and cisplatin (5 μmol / L) were used together was 469.3 nmol / L. As described above, compound A significantly enhanced the anti-tumor effect of cisplatin. It is believed that its effect is greater than gemcitabine, which is an existing drug.

[0272] The CI when cisplatin and gemcitabine were used together was 0.78, and the CI when cisplatin and compound A were used together was 0.13. CI < 1, so a synergistic effect based on the combined use of compound A and cisplatin was confirmed. In addition, it is estimated that the smaller the CI value, the higher the synergistic effect, so it can be said that the synergistic effect of compound A is more significant than gemcitabine, which is an existing drug.

[0273] (Test Example 5)

[0274] Evaluation of the antitumor activity of cisplatin-based combination therapy against cholangiocarcinoma cell line HuCC-T1 in spheroid culture

[0275] As test substances, gemcitabine, cisplatin, and mesylate of compound A were used.

[0276] Gemcitabine was prepared by dissolving gemcitabine hydrochloride (manufactured by Plantex) in dimethyl sulfoxide (DMSO). Cisplatin (manufactured by Wako Pure Chemical Industries, Ltd., Cat. #039-20091) was dissolved in DMSO.

[0277] HuCC-T1 cells, a cholangiocarcinoma cell line, were subcultured using RPMI-1640 medium containing 10% serum (Thermo Fisher Scientific Inc., Cat. #10437-028). 2 Incubator (37°C, 5% CO 2 The cells were diluted to 5,000 cells / well / 100 μL using a 10% serum medium and seeded in an Ultra-Low Adhesive 96-well Plate (Cat. #7007 manufactured by Corning Incorporated). Spheroids were formed after 4 days of culture.

[0278] The mesylate of compound A and Gemcitabine were dissolved in DMSO to prepare 10mmol / L DMSO solutions. The solutions were diluted with DMSO in sequence to prepare DMSO solutions with a concentration of 1000 times the final treatment concentration. Cisplatin was dissolved in DMSO to prepare a 100mmol / L DMSO solution. The solutions were diluted with DMSO to prepare a DMSO solution with a concentration of 5000 times the final treatment concentration. Furthermore, the DMSO solution of Cisplatin was diluted with 10% serum medium to prepare a medium containing Cisplatin with a concentration of 10 times the final treatment concentration. The DMSO diluted solutions of compound A and Gemcitabine were diluted with the medium containing Cisplatin to prepare treatment solutions with a concentration of 10 times the final treatment concentration. For compound A and Gemcitabine, the maximum concentration was set to 10μmol / L, and 9 concentrations were combined with Cisplatin (10μmol / L) at a common ratio of 1 / 3 and used.

[0279] 10 μL of compound A or Gemcitabine diluted with cisplatin-containing culture medium was added to each well. In addition, a group in which only solvent without drug was added to the wells seeded with cells (positive control group) and a group in which only solvent without drug was added to the wells added with culture medium (negative control group) were set up. Both were set to n = 3 wells.

[0280] After adding the drug, the cells were cultured for 3 days, and the cell viability was evaluated using CellTiter Glo (registered trademark) Reagent (Cat. #G7570 manufactured by PROMEGA Co.) using the intracellular ATP level as an indicator. The concentration that inhibited the cell viability by 50% (IC50 value) was calculated using XLFit software Ver.3 (registered trademark) (manufactured by CTC).

[0281] The cell survival rate of each well was determined by setting the luminescence signal amount of the negative control group as 0% cell survival rate and the luminescence signal amount of the positive control group as 100%. The mean value and standard deviation of the cell survival rate of each treatment group were calculated to prepare Table 10.

[0282] Results of combined use test with Cisplatin 10μmol / L

[0283] [Table 10]

[0284] The average cell survival rate

[0285] Concentration (nM) 10000.0 3000.0 1000.0 300.0 100.0 30.0 10.0 3.0 1.0 Gemcitabine 50 73 79 84 88 90 94 96 94 Compound A 34 35 36 41 58 77 87 93 95

[0286] Standard deviation of cell viability

[0287] Concentration (nM) 10000.0 3000.0 1000.0 300.0 100.0 30.0 10.0 3.0 1.0 Gemcitabine 5.9 1.3 0.5 3.5 4.5 2.3 3.3 4.0 2.4 Compound A 2.8 2.1 2.3 5.8 1.7 1.4 6.3 5.8 1.7

[0288] Furthermore, the graph produced according to Table 10 is shown in Fig. 9 middle.

[0289] The IC50 value when gemcitabine and cisplatin (10 μmol / L) were used together in spheroid culture was 10000 nmol / L or more, and the IC50 value when compound A and cisplatin (10 μmol / L) were used together was 164.6 nmol / L. As described above, compound A significantly enhanced the anti-tumor effect of cisplatin. It is believed that its effect is greater than gemcitabine, which is an existing drug.

[0290] The CI when cisplatin and gemcitabine were used together was 0.79, and the CI when cisplatin and compound A were used together was 0.42. CI < 1, so a synergistic effect based on the combined use of compound A and cisplatin was confirmed. In addition, it is estimated that the smaller the CI value, the higher the synergistic effect, so it can be said that the synergistic effect of compound A is more significant than gemcitabine, which is an existing drug.

[0291] (Test Example 6)

[0292] Evaluation of the antitumor activity of cisplatin-based combinations against breast cancer cell line HCC1954 in spheroid culture

[0293] As test substances, gemcitabine, cisplatin, and mesylate of compound A were used.

[0294] Gemcitabine was prepared by dissolving gemcitabine hydrochloride (manufactured by Plantex) in dimethyl sulfoxide (DMSO). Cisplatin (manufactured by Wako Pure Chemical Industries, Ltd., Cat. #039-20091) was dissolved in DMSO.

[0295] HCC1954 cells, a human breast cancer cell line, were subcultured using RPMI-1640 medium containing 10% serum (Thermo Fisher Scientific Inc., Cat. #10437-028). In this experiment, all cells were cultured under CO 2 Incubator (37°C, 5% CO 2 The cells were diluted to 5,000 cells / well / 100 μL using a 10% serum medium and seeded in an Ultra-Low Adhesive 96-well Plate (Cat. #7007 manufactured by Corning Incorporated). Spheroids were formed after 4 days of culture.

[0296] The mesylate of compound A and Gemcitabine were dissolved in DMSO to prepare 10mmol / L DMSO solutions. The solutions were diluted with DMSO in sequence to prepare DMSO solutions with a concentration of 1000 times the final treatment concentration. Cisplatin was dissolved in DMSO to prepare a 100mmol / L DMSO solution. The solutions were diluted with DMSO to prepare a DMSO solution with a concentration of 5000 times the final treatment concentration. Furthermore, the DMSO solution of Cisplatin was diluted with 10% serum medium to prepare a medium containing Cisplatin with a concentration of 10 times the final treatment concentration. The DMSO diluted solutions of compound A and Gemcitabine were diluted with the medium containing Cisplatin to prepare treatment solutions with a concentration of 10 times the final treatment concentration. For compound A and Gemcitabine, the maximum concentration was set to 10μmol / L, and 9 concentrations were combined with Cisplatin (10μmol / L, 5μmol / L) at a common ratio of 1 / 3 and used.

[0297] 10 μL of compound A or Gemcitabine diluted with cisplatin-containing culture medium was added to each well. In addition, a group in which only solvent without drug was added to the wells seeded with cells (positive control group) and a group in which only solvent without drug was added to the wells added with culture medium (negative control group) were set up. Both were set to n = 3 wells.

[0298] After adding the drug, the cells were cultured for 3 days, and the cell viability was evaluated using CellTiter Glo (registered trademark) Reagent (Cat. #G7570 manufactured by PROMEGA Co.) using the intracellular ATP level as an indicator. The concentration that inhibited the cell viability by 50% (IC50 value) was calculated using XLFit software Ver.3 (registered trademark) (manufactured by CTC).

[0299] The cell survival rate of each well was determined by setting the luminescence signal amount of the negative control group as 0% cell survival rate and the luminescence signal amount of the positive control group as 100%. The mean value and standard deviation of the cell survival rate of each treatment group were calculated to prepare Table 11.

[0300] Results of combined use test with Cisplatin 10μmol / L

[0301] [Table 11]

[0302] The average cell survival rate

[0303] Concentration (nM) 10000.0 3000.0 1000.0 300.0 100.0 30.0 10.0 3.0 1.0 Gemcitabine 50 52 60 73 88 97 101 100 96 Compound A 2 2 2 15 38 65 75 94 92

[0304] Standard deviation of cell viability

[0305] Concentration (nM) 10000.0 3000.0 1000.0 300.0 100.0 30.0 10.0 3.0 1.0 Gemcitabine 2.2 17.3 7.3 6.9 3.1 3.5 1.7 2.7 4.4 Compound A 2.2 1.5 1.9 3.0 2.2 7.5 8.6 7.4 10.9

[0306] Furthermore, the graph prepared according to Table 11 is shown in Fig.10 middle.

[0307] The IC50 value when gemcitabine and cisplatin (10 μmol / L) were used together in spheroid culture was 10000 nmol / L or more, and the IC50 value when compound A and cisplatin (10 μmol / L) were used together was 49.4 nmol / L. As described above, compound A significantly enhanced the anti-tumor effect of cisplatin. It is believed that its effect is greater than gemcitabine, which is an existing drug.

[0308] The CI when cisplatin and gemcitabine were used together was 0.60, and the CI when cisplatin and compound A were used together was 0.03. CI < 1, so a synergistic effect based on the combined use of compound A and cisplatin was confirmed. In addition, it is estimated that the smaller the CI value, the higher the synergistic effect, so it can be said that the synergistic effect of compound A is more significant than gemcitabine, which is an existing drug.

[0309] (Test Example 7)

[0310] Experimental study on the combined effect of cholangiocarcinoma cell lines in subcutaneous transplanted tumor-bearing mice

[0311] As test substances, gemcitabine and the mesylate of compound A were used.

[0312] As gemcitabine, a product prepared by dissolving gemcitabine hydrochloride (manufactured by TEVA) in physiological saline was used, and as cisplatin, a product prepared by dissolving cisplatin (manufactured by Wako Pure Chemical Industries, Ltd., Cat. #039-20091) in physiological saline was used.

[0313] TFK-1 cells or HuCCT-1 cells, which are human cholangiocarcinoma cell lines, were subcutaneously injected into the posterior abdomen of female BALB / cA Jcl-nu mice at 5 to 6 weeks of age. After tumor transplantation, the long diameter (mm) and short diameter (mm) of the tumor were measured to calculate the tumor volume (TV). Mice were allocated to each group in such a way that the average TV of each group became equal, and the day of the grouping was set as day 1.

[0314] The dosage of cisplatin was set by referring to information retrieval and Cancer Genomics & Proteomics, 2012, Volume 9, pages 77-92 (non-patent document 6), and the dosage of gemcitabine and compound A was set by referring to non-patent document 6 and The journal of pharmacology and experimental therapeutics, 2018, Volume 366, pages 125-135 (non-patent document 7). The test solution of the cisplatin alone group was prepared to 1-10 mg / kg / day as the dosage. In addition, the test solution of the compound A alone group was prepared to 30-240 mg / kg / day. Compound A was supplied from the tail vein of mice once a week for a total of 3 times from day 1, and cisplatin was supplied by intraperitoneal administration to mice once a week for a total of 3 times from day 1. In the combined administration group, Compound A was administered at 30 to 240 mg / kg / day and cisplatin was administered at 1 to 10 mg / kg / day.

[0315] As a comparative experiment, gemcitabine was used as a control drug. The test solution of the gemcitabine alone group was prepared at 30-240 mg / kg / day. Gemcitabine was administered from the tail vein of mice once a week for a total of 3 times starting from day 1, and in the combined administration group, gemcitabine was administered at 30-240 mg / kg / day and cisplatin was administered at 1-10 mg / kg / day.

[0316] In this test, the dosage of compound A and gemcitabine was set to use the MTD of each drug. Cisplatin was used in the maximum dosage that can be used with each drug. Regarding anti-tumor agents, the dosage that shows the maximum efficacy is very close to the dosage that toxicity appears. In order to evaluate the maximum anti-tumor effect of the drug with an animal model, it is generally evaluated near the MTD. In this test example, the definition of MTD and the maximum effect dosage is almost the same.

[0317] As an indicator of antitumor effect, TV in each drug administration group was measured, and the relative tumor volume (RTV) and T / C (%) relative to day 1 were calculated by the following formula to evaluate the antitumor effect. Regarding the evaluation and judgment of the combined effect, if the average RTV value of the combined administration group was statistically significantly (Welch's IUT, over all maximum p < 0.05) lower than the average RTV value of each single administration group, it was judged that there was a combined effect.

[0318] TV(mm 3 )=(long diameter×short diameter 2 ) / 2

[0319] RTV = (TV on the day of tumor measurement) / (TV on Day 1)

[0320] T / C (%) = [(average RTV value of the test solution administration group) / (average RTV value of the control group)] × 100

[0321] (Test Example 8)

[0322] Study on the effect of compound A combined with antitumor platinum complex in cancer patients

[0323] <Preparation of liquid pharmaceutical composition>

[0324] The methanesulfonate of Compound A was dissolved in an appropriate amount of water for injection, and the pH was adjusted using a 1 mol / L sodium hydroxide aqueous solution. An appropriate amount of water for injection was added and mixed so that the concentration of Compound A became 20 mg / mL.

[0325] Furthermore, glycerol (manufactured by Merck KGaA, molecular weight 92) was added to a concentration of 1.5% by mass. The pH of the liquid pharmaceutical preparation was 2.9, and the liquid was filtered using a membrane filter (0.22 μm) to obtain a liquid pharmaceutical preparation.

[0326] <Administration of medication and determination of therapeutic effects>

[0327] The following medication cycle is repeated, i.e., for cancer patients, compound A and cisplatin are administered once a week by intravenous injection from the first to second week, and no administration is performed in the third week. Specifically, a 21-day cycle is repeated, i.e., compound A and cisplatin are administered on the first and eighth days, with 21 days as one cycle. The dosage of compound A per administration is set to 8 to 135 mg / m 2 The dosage of cisplatin is set at 10 to 50 mg / m 2 .

[0328] The effectiveness of treatment can be determined by the following criteria.

[0329] The evaluation subjects were confirmed by image diagnosis using MRI (magnetic resonance imaging), and the evaluation was performed according to the following criteria.

[0330] CR (Complete Response): The state where the tumor completely disappears

[0331] PR (Partial Response): The sum of tumor size has decreased by more than 30%.

[0332] SD (Stable Disease): A state in which the tumor size does not change

[0333] PD (Progressive Disease): The sum of the tumor sizes increases by more than 20% and the absolute value increases by more than 5 mm, or new lesions appear.

[0334] Industrial Applicability

[0335] The present invention is useful as an antitumor agent, an antitumor kit, and an antitumor effect enhancer that exhibit a significant antitumor effect.

Claims

1. Use of a combination of cisplatin and 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof in the manufacture of an antitumor agent, The tumor is biliary tract cancer, breast cancer or pancreatic cancer.

2. The use according to claim 1, wherein The bile duct cancer is cholangiocarcinoma.

3. Use of a combination of a preparation containing cisplatin and a preparation containing 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof in the manufacture of an antitumor kit, The tumor is biliary tract cancer, breast cancer or pancreatic cancer.

4. The use according to claim 3, wherein The bile duct cancer is cholangiocarcinoma.

5. Use of a combination of oxaliplatin and 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine or a salt thereof in the manufacture of an antitumor agent for pancreatic cancer.

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