Combination drugs for cancer treatment

Abstract

A novel combination drug for cancer treatment is provided. The present invention relates to a combination drug for cancer treatment, comprising a tyrosine kinase inhibitor and Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof.

Description

[Technical field]

[0001] The present invention relates to a pharmaceutical combination for treating cancer. [Background technology]

[0002] Probiotics are defined as "live microorganisms that have a positive effect on the health of the host by improving the balance of intestinal flora," and efforts are being actively made to apply them in practical clinical applications in the medical field.

[0003] Probiotics are also expected to enhance the therapeutic effects of cancer patients when used in combination with chemotherapy, such as anticancer drugs.

[0004] For example, Patent Document 1 describes that the therapeutic effect of an immune checkpoint inhibitor can be enhanced by administering the immune checkpoint inhibitor and Clostridium butyricum MIYAIRI 588 (FERM BP-2789) to a cancer patient. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2021 / 216618 Summary of the Invention

[0006] An object of the present invention is to provide a novel combination drug for treating cancer.

[0007] The present inventors have surprisingly found that the antitumor effect can be improved by combining Clostridium butyricum MIYAIRI 588 (FERM BP-2789) with a tyrosine kinase inhibitor. Based on this finding, the present invention has been completed.

[0008] That is, according to one embodiment of the present invention, there is provided a combination drug for cancer treatment comprising a tyrosine kinase inhibitor and Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof. [Brief description of the drawings]

[0009] [Figure 1] 1 is a graph showing the time course of tumor volume in each group in Test Example 1. [Diagram 2] 1 is a graph showing the tumor volume at each observation time point (4th day, 8th day, 11th day, 15th day, and 18th day) in each group of Test Example 1. [Diagram 3] 1 is a graph showing a Kaplan-Meier curve in Test Example 1, with a tumor volume of 1000 mm3 as an event. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] An embodiment according to one aspect of the present invention will be described below, but the present invention is not limited to the following embodiment.

[0011] In this specification, the range "X to Y" means "X or more and Y or less." Furthermore, unless otherwise specified, the operations and measurements of physical properties are performed under conditions of room temperature (20 to 25°C) and relative humidity of 40 to 50% RH.

[0012] In this specification, "Clostridium butyricum MIYAIRI 588 (FERM BP-2789)" is also referred to simply as "CBM588".

[0013] A first aspect of the present invention is a combination pharmaceutical for treating cancer, comprising a tyrosine kinase inhibitor and CBM588 or a culture thereof.

[0014] A second aspect of the present invention is a method for treating cancer, comprising administering to a subject in combination an effective amount of a tyrosine kinase inhibitor and an effective amount of CBM588 or a culture thereof.

[0015] A third aspect of the invention is a pharmaceutical composition comprising a tyrosine kinase inhibitor in combination with CBM588 or a culture thereof for use in the treatment of cancer.

[0016] A fourth aspect of the present invention is a pharmaceutical composition for treating cancer, comprising CBM588 or a culture thereof administered in combination with a tyrosine kinase inhibitor.

[0017] A fifth aspect of the present invention is a pharmaceutical composition for treating cancer comprising a tyrosine kinase inhibitor administered in combination with CBM588 or a culture thereof.

[0018] A sixth aspect of the present invention is a kit for treating cancer, comprising a tyrosine kinase inhibitor and CBM588 or a culture thereof.

[0019] A seventh aspect of the invention is CBM588 or a culture thereof for use in combination with a tyrosine kinase inhibitor in a method of treating cancer.

[0020] An eighth aspect of the invention is a tyrosine kinase inhibitor for use with CBM588, or a culture thereof, in a method of treating cancer.

[0021] A ninth aspect of the invention is the use of a tyrosine kinase inhibitor and / or CBM588 or a culture thereof in a medicament for use in a method of treating cancer comprising administering to a subject a tyrosine kinase inhibitor and CBM588 or a culture thereof.

[0022] In the present invention, by administering a tyrosine kinase inhibitor in combination with CBM588 or a culture thereof, the anti-tumor effect, particularly the tumor volume reduction effect, can be improved compared to the administration of a tyrosine kinase inhibitor alone, thereby significantly increasing the survival rate of the subject.

[0023] The tyrosine kinase inhibitor of the present invention is used in combination with Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof for the treatment of cancer.

[0024] Tyrosine kinase inhibitors are drugs that exert their antitumor effects by blocking cellular tyrosine kinases, enzymes involved in many cellular functions, including cell signaling, growth, and division.

[0025] The tyrosine kinase inhibitor according to the present invention is not particularly limited, and any known tyrosine kinase inhibitor can be used. The tyrosine kinase inhibitor is preferably at least one selected from the group consisting of EGFR tyrosine kinase inhibitors, c-kit receptor tyrosine kinase inhibitors, BCR-ABL tyrosine kinase inhibitors, and multikinase inhibitors, and more preferably a multikinase inhibitor.

[0026] Specific examples of tyrosine kinase inhibitors according to the present invention are not particularly limited, but include at least one selected from the group consisting of imatinib, dasatinib, nilotinib, afatinib, erlotinib, osimertinib, gefitinib, axitinib, cabozantinib, sunitinib, sorafenib, pazopanib, regorafenib, and lenvatinib, and more preferably at least one selected from the group consisting of axitinib, cabozantinib, sunitinib, sorafenib, pazopanib, regorafenib, and lenvatinib.

[0027] The Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof according to the present invention is used in combination with a tyrosine kinase inhibitor for the treatment of cancer.

[0028] CBM588 is a spore-forming bacteria that is resistant to various external environments when in the spore state. Therefore, when CBM588 is orally administered to humans or animals in the form of spores, CBM588 does not die completely even when it comes into contact with digestive fluids such as gastric acid, intestinal fluid, and bile acid, but can reach the fermentation sites from the lower small intestine to the large intestine and continue to grow.

[0029] Furthermore, CBM588 is widely available commercially as a probiotic, feed additive, and food, and has been shown to cause no side effects even when administered over long periods of time to humans, livestock, and other mammals, ensuring its high safety.

[0030] CBM588 was deposited on May 1, 1981 at the Microbial Industrial Technology Institute, Agency of Industrial Science and Technology, Ministry of International Trade and Industry (currently the Patent Organism Depositary Center, National Institute of Technology and Evaluation, Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan) under the accession number FERM BP-2789, and transferred to an international depositary organization under the Budapest Treaty on March 6, 1990, where it is deposited under the accession number FERM BP-2789.

[0031] CBM588 is commercially available as a live bacteria preparation from Miyarisan Pharmaceutical Co., Ltd., and has no side effects even when administered to humans or animals for a long period of time. In a preferred embodiment, CBM588 is a live bacteria.

[0032] In the present invention, the culture of CBM588 refers to a culture medium in which Clostridium butyricum is cultured, a residue containing bacteria obtained by centrifuging the culture medium, and a dried product of the residue.

[0033] The culture of CBM588 can be obtained by a known method for culturing microorganisms, for example, the method disclosed in JP-A-08-252088. One embodiment of the method is shown below: CBM588 is cultured in a medium containing 1.0 (w / v)% peptone, 1.0 (w / v)% yeast extract, 1.0 (w / v)% corn starch and 0.2 (w / v)% precipitated calcium carbonate for 10 min. 5 ~10 6 The resulting culture solution is inoculated at a concentration of 1000 to 6000 g / mL and then statically cultured at 37° C. for 48 hours to obtain a "culture solution of CBM588". Next, the resulting culture solution is centrifuged (2,000 to 6,000 g x 10 to 30 minutes) to separate the "residue containing bacteria obtained by centrifuging the culture solution", and the residue is dried at 0 to 80° C., preferably 10 to 20° C., for 1 to 24 hours, preferably 5 to 18 hours, by air drying, or the like, or at 0 to 80° C., preferably 10 to 20° C., at 0.05 to 500 Torr (7 Pa to 66.7 kPa), preferably 1 to 100 Torr (133 Pa to 13.3 kPa), for 1 to 24 hours, preferably 2 to 15 hours, to obtain a "dried product of the residue". To obtain the dried product, spray drying, freeze drying, or the like may be used.

[0034] The medium used for culturing CBM588 of the present invention may be either a synthetic medium or a natural medium, so long as it contains a carbon source that can be assimilated by CBM588, an appropriate amount of a nitrogen source, inorganic salts, and other nutrients such as vitamins.

[0035] For example, the carbon source used in the medium according to the present invention is not particularly limited as long as it is a carbon source that can be assimilated by CBM588. The carbon source is not necessarily limited to sugar, but in consideration of the growth of the bacterial cells, a sugar or a sugar-containing source that can be used by CBM588 is preferably used. Specific examples of usable carbon sources include cellobiose, glucose, fructose, galactose, lactose, maltose, mannose, melibiose, raffinose, salicin, starch, sucrose, trehalose, xylose, dextrin, and molasses, taking assimilation into consideration. Among these carbon sources, starch, glucose, fructose, sucrose, and molasses are preferably used. One or more of the above carbon sources may be selected and used in consideration of CBM588. In this case, the concentration of the carbon source added varies depending on the type of carbon source used and the medium composition other than the carbon source of the medium used, but is usually 0.5 to 5 (w / v)%, preferably 2 to 4 (w / v)%.

[0036] Examples of nitrogen sources and vitamins include meat extract, peptone, yeast extract, soybean and wheat hydrolysates such as seasoning liquid, soybean powder, milk casein, casamino acid, various amino acids, corn steep liquor, organic nitrogen compounds such as hydrolysates of other animals, plants and microorganisms, and ammonium salts such as ammonium sulfate. Among these nitrogen sources, peptone, yeast extract, meat extract, corn steep liquor and seasoning liquid are preferably used. One or more of the above-mentioned nitrogen sources and vitamins may be selected and used to improve the growth of CBM588. In this case, the concentration of the nitrogen source added varies depending on the type of strain and nitrogen source used, and the medium composition other than the nitrogen source of the medium used, but when peptone containing a large amount of nitrogen source is used, the concentration is usually 0.5 to 4 (w / v)%, preferably 1 to 3 (w / v)%. When seasoning liquid or corn steep liquor containing a large amount of nitrogen source and vitamins is used, the concentration is usually 0.5 to 5 (w / v)%, preferably 1 to 4 (w / v)%. Furthermore, when yeast extract or meat extract containing a large amount of vitamins is used, the concentration is usually 0.5 to 4 (w / v)%, preferably 1 to 3 (w / v)%.

[0037] Furthermore, as the inorganic salt, one or more selected from phosphates, hydrochlorides, sulfates, butyrates, propionates, acetates, etc. of magnesium, manganese, calcium, sodium, potassium, molybdenum, strontium, boron, copper, iron, tin, zinc, etc. may be used. In addition, antifoaming agents, vegetable oils, surfactants, blood and blood components, drugs such as antibiotics, physiologically active substances such as plant or animal hormones, etc. may be appropriately added to the medium as necessary.

[0038] Since CBM588 is obligately anaerobic in the present invention, the culture conditions must be such that the culture is performed under anaerobic conditions without aeration, while aerating nitrogen or carbon dioxide, or by adding a reducing agent to the medium to lower the redox potential. The culture conditions are appropriately selected depending on the composition of the medium and the culture method used, and are not particularly limited as long as the conditions allow CBM588 to grow. Specifically, the culture temperature is usually 20 to 42°C, preferably 35 to 40°C.

[0039] In the present invention, since the growth of CBM588 is promoted by neutralizing the acid produced during the culture with an alkali, it is preferable to add calcium carbonate to the medium in advance. In this case, the amount of calcium carbonate added is usually 0.1 to 4 (w / v)%, preferably 0.2 to 2.5 (w / v)%. Alternatively, it is also preferable to carry out the neutralization step while controlling the pH of the medium within a set pH range using an alkaline aqueous solution such as sodium hydroxide, sodium bicarbonate, sodium carbonate, potassium hydroxide, and potassium carbonate. In addition, when an alkaline aqueous solution is used, the "set pH" means the pH of the medium that is set in advance during the culture period, and the "set pH range" means the pH range that is acceptable during the culture period, and is generally expressed as the set pH ± tolerance. According to the present invention, the set pH is usually set within a range of 5.0 to 7.5, preferably 5.5 to 6.5, and the set pH range is the set pH ± 0.5, desirably the set pH ± 0.2.

[0040] In the present invention, the pH of the medium during the culture is near neutral, more preferably 6.5 to 7.5, when inoculating CBM588. When using an alkaline aqueous solution, it is preferable to maintain the pH within the set range while gently stirring to prevent oxygen from being mixed in. By controlling the pH during inoculation and growth of CBM588 in this way, the bacterial density can be dramatically increased.

[0041] In the culture according to the present invention, the initial culture concentration of CBM588 is not particularly limited as long as it is within a range in which CBM588 can grow. Specifically, it is usually 10 4 ~10 7 / mL, preferably 10 5 ~10 6 pieces / mL.

[0042] The thus obtained CBM588 culture can be used in combination with a tyrosine kinase inhibitor to improve the antitumor effect.

[0043] In the present invention, a tyrosine kinase inhibitor and CBM588 or a culture thereof may be used in combination with an immune checkpoint inhibitor to treat cancer.

[0044] Immune checkpoint inhibitors inhibit the function of immune checkpoint receptors or ligands, and examples of such inhibitors include antagonists for inhibitory receptors and agonists for costimulatory immune checkpoint receptors.

[0045] The term "antagonist" includes various substances that block receptor activation through binding between the receptor and a ligand, such as substances that bind to the receptor and block the receptor-ligand binding, and substances that bind to the ligand and block the receptor-ligand binding.

[0046] Examples of antagonists against inhibitory immune checkpoints include antagonistic antibodies that bind to inhibitory immune checkpoint molecules (inhibitory receptors or ligands for such receptors), soluble polypeptides that do not activate receptors and are designed based on inhibitory immune checkpoint ligands, or vectors capable of expressing such polypeptides.

[0047] The immune checkpoint inhibitor according to the present invention is not particularly limited, and any conventionally known immune checkpoint inhibitor can be used. The immune checkpoint inhibitor is preferably at least one selected from the group consisting of PD-1 inhibitors, PD-L1 inhibitors, and CTLA-4 inhibitors, and more preferably a PD-1 inhibitor.

[0048] Specific examples of immune checkpoint inhibitors according to the present invention are not particularly limited, but include at least one selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, semipilimab, durvalumab, daclizumab, avelumab, and atezolizumab, and more preferably at least one selected from the group consisting of nivolumab, pembrolizumab, and semipilimab.

[0049] The present invention includes the following embodiments: A combination drug for treating cancer comprising a tyrosine kinase inhibitor, CBM588 and an immune checkpoint inhibitor; A method for treating cancer, comprising administering to a subject in combination an effective amount of a tyrosine kinase inhibitor, an effective amount of CBM588 or a culture thereof, and an effective amount of an immune checkpoint inhibitor; A pharmaceutical composition comprising a combination of a tyrosine kinase inhibitor, CBM588 or a culture thereof, and an immune checkpoint inhibitor for use in the treatment of cancer; a pharmaceutical composition for treating cancer, comprising CBM588 or a culture thereof administered in combination with a tyrosine kinase inhibitor and an immune checkpoint inhibitor; a pharmaceutical composition for treating cancer comprising CBM588 or a culture thereof and a tyrosine kinase inhibitor administered in combination with an immune checkpoint inhibitor; A kit for cancer treatment comprising a tyrosine kinase inhibitor, CBM588 or a culture thereof, and an immune checkpoint inhibitor; CBM588 or a culture thereof for use with a tyrosine kinase inhibitor and an immune checkpoint inhibitor in a method for treating cancer; and A tyrosine kinase inhibitor for use in combination with CBM588 or a culture thereof and an immune checkpoint inhibitor in a method for treating cancer.

[0050] In the present invention, the type of "cancer" to be treated is not particularly limited. Examples of cancer include oral cancer, pharyngeal cancer, esophageal cancer, gastric cancer, liver cancer, gallbladder cancer, biliary tract cancer, spleen cancer, colon cancer, small intestine cancer, duodenal cancer, colon cancer, colon adenocarcinoma, rectal cancer, pancreatic cancer, liver cancer, bladder cancer, renal cell cancer, non-small cell lung cancer, small cell lung cancer, malignant melanoma (melanoma), sarcoma, lymphoma, breast cancer, cervical cancer, uterine cancer, ovarian cancer, testicular cancer, prostate cancer, head and neck cancer, and metastatic cancers thereof. The cancer is preferably at least one selected from the group consisting of colon cancer, metastatic renal cell carcinoma (mRCC), non-small cell lung cancer, melanoma, sarcoma, lymphoma, breast cancer, bladder cancer, cervical cancer, head and neck cancer, liver cancer, gastric cancer, and rectal cancer.

[0051] In the present invention, the term "effective amount" means at least the amount of an active ingredient (i.e., a tyrosine kinase inhibitor, CBM588, or an immune checkpoint inhibitor) required to exert a desired effect such as the treatment of cancer, particularly an improved antitumor effect.

[0052] In the present invention, the "subject" is not particularly limited, but it is a mammal or a bird, preferably a mammal or a bird that is or may be affected by cancer. Here, the mammal includes both primates such as humans, monkeys, gorillas, chimpanzees, and orangutans, as well as non-human mammals such as mice, rats, hamsters, guinea pigs, rabbits, dogs, cats, pigs, cows, horses, sheep, camels, and goats. Birds include chickens, quails, and pigeons. Of these, humans are preferred.

[0053] In the present invention, a tyrosine kinase inhibitor, CBM588 or a culture thereof, and optionally an immune checkpoint inhibitor, are administered in combination (co-administered) to a subject.

[0054] Administration in combination (concomitant administration) includes simultaneous administration of each component, and sequential administration of each component at predetermined intervals over a treatment period (combination therapy). The route and means of administration of each component administered in combination may be the same or different.

[0055] The dosage and administration method of the tyrosine kinase inhibitor, CBM588 or a culture thereof, and the immune checkpoint inhibitor can be determined appropriately taking into consideration the type of cancer, the symptoms, age, sex, weight, and condition of the subject, etc.

[0056] The dosage of the tyrosine kinase inhibitor is, for example, about 1 to about 2400 mg / kg body weight / day as an active ingredient. The tyrosine kinase inhibitor can be administered 1 to 6 times a day, every day, every other day, or every few days. The method of administration of the tyrosine kinase inhibitor is not particularly limited, and includes oral administration, intravenous injection, intraarterial injection, subcutaneous injection, intradermal injection, intraperitoneal injection, intramuscular injection, intrathecal injection, parenteral administration such as transdermal administration or transdermal absorption, and the like, and is preferably oral administration. In the examples described below, the dosage of the tyrosine kinase inhibitor is 3 mg / kg body weight / day (5 times a week), and the administration method is oral administration.

[0057] The dosage of CBM588 or a culture thereof is, for example, 0.1 to 1000 mg / kg body weight / day as the active ingredient. CBM588 or a culture thereof can be administered 1 to 6 times per day, every day, every other day, or every few days. The method of administration of CBM588 or a culture thereof is not particularly limited, and examples thereof include oral administration, intravenous injection, intraarterial injection, subcutaneous injection, intradermal injection, intraperitoneal injection, intramuscular injection, intrathecal injection, transdermal administration, transdermal absorption, and other parenteral administration, and oral administration is preferred. In the examples described below, the dosage of CBM588 is about 43 mg / kg body weight / day (1×10 8 cfu (approximately 1.3 mg) / 100 μL / animal), and the method of administration is oral administration.

[0058] The dosage of the immune checkpoint inhibitor is, for example, 0.1 to 20 mg / kg body weight / day as an active ingredient, preferably 2 to 3 mg / kg body weight / day. The immune checkpoint inhibitor can be administered 1 to 6 times per day, every day, every other day, or every few days. The method of administration of the immune checkpoint inhibitor is not particularly limited, and includes oral administration, intravenous injection, intraarterial injection, subcutaneous injection, intradermal injection, intraperitoneal injection, intramuscular injection, intrathecal injection, parenteral administration such as percutaneous administration or percutaneous absorption, and preferably intraperitoneal administration or intravenous injection. In the examples described later, the dosage of the immune checkpoint inhibitor is 10 mg / kg body weight / day (5 times every 3 days), and the administration method is intraperitoneal administration.

[0059] In the present invention, the tyrosine kinase inhibitor, CBM588 or a culture thereof, and the immune checkpoint inhibitor may be in the form of a combined medicine, or each may be in the form of a separate medicine (or pharmaceutical composition).

[0060] The combination medicine may be in the form of a combination preparation containing each component in the same composition, or each component may be prepared separately and in the form of a single package suitable for combined administration (i.e., in the form of a kit).

[0061] In the present invention, in addition to the above-mentioned components, additives acceptable for formulation can be further used, such as excipients, stabilizers, preservatives, wetting agents, emulsifiers, lubricants, sweeteners, colorants, flavors, buffers, antioxidants, pH adjusters, binders, thickeners, dispersants, suspending agents, disintegrants, bacteriostatic agents, surfactants, etc.

[0062] In the present invention, the dosage forms of the combination medicine, pharmaceutical composition, tyrosine kinase inhibitor, CBM588 or a culture thereof, and immune checkpoint inhibitor are not particularly limited and may be set as appropriate, and examples thereof include tablets, powders, fine granules, granules, capsules, pills, sustained release agents, solutions, suspensions, emulsions, lotions, injections, drip infusions, topical agents, suppositories, and patches.

[0063] The following are exemplary embodiments of the present invention. [1] A combination drug for cancer treatment comprising a tyrosine kinase inhibitor and Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof. [2] The combination drug for cancer treatment described in [1], wherein the Clostridium butyricum miyairi 588 is a live bacterium. [3] The combination drug for cancer treatment according to [1] or [2], wherein the tyrosine kinase inhibitor is at least one selected from the group consisting of an EGFR tyrosine kinase inhibitor, a c-kit receptor tyrosine kinase inhibitor, a BCR-ABL tyrosine kinase inhibitor, and a multikinase inhibitor. [4] The combination drug for cancer treatment according to any one of [1] to [3], wherein the tyrosine kinase inhibitor is at least one selected from the group consisting of imatinib, dasatinib, nilotinib, afatinib, erlotinib, osimertinib, gefitinib, axitinib, cabozantinib, sunitinib, sorafenib, pazopanib, regorafenib, and lenvatinib. [5] The combination drug for cancer treatment according to any one of [1] to [4], further comprising an immune checkpoint inhibitor. [6] The combination drug for cancer treatment according to [5], wherein the immune checkpoint inhibitor is at least one selected from the group consisting of a PD-1 inhibitor, a PD-L1 inhibitor, and a CTLA-4 inhibitor. [7] The combination drug for cancer treatment according to [5] or [6], wherein the immune checkpoint inhibitor is at least one selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, semipilimab, durvalumab, daclizumab, avelumab, and atezolizumab. [8] The combination drug for cancer treatment according to any of [1] to [7], wherein the cancer is at least one selected from the group consisting of colorectal cancer, metastatic renal cell carcinoma (mRCC), non-small cell lung cancer, melanoma, sarcoma, lymphoma, breast cancer, bladder cancer, cervical cancer, head and neck cancer, liver cancer, gastric cancer, and rectal cancer. [9] A method for treating cancer, comprising administering to a subject in combination an effective amount of a tyrosine kinase inhibitor and an effective amount of Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof.

[10] A pharmaceutical composition comprising a combination of a tyrosine kinase inhibitor and Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof for use in treating cancer.

[11] A pharmaceutical composition for cancer treatment, comprising Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof, administered in combination with a tyrosine kinase inhibitor.

[12] A pharmaceutical composition for treating cancer, comprising a tyrosine kinase inhibitor administered in combination with Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof.

[13] A cancer treatment kit comprising a tyrosine kinase inhibitor and Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof.

[14] Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof for use together with a tyrosine kinase inhibitor in a method for treating cancer.

[15] A tyrosine kinase inhibitor for use together with Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof in a method for treating cancer.

[16] Use of a tyrosine kinase inhibitor and / or Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof in a medicament for use in a method for treating cancer comprising administering to a subject a tyrosine kinase inhibitor and Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof. EXAMPLES

[0064] The present invention will be described below using specific examples, but the present invention is not limited to these examples. Unless otherwise specified, the operations were carried out at room temperature (25° C.).

[0065] Animal experiments using mice were conducted with the approval of the Animal Experimentation Committee of the Osaka International Cancer Institute (approval number: 23022121).

[0066] <Test Example 1> BALB / c mice (7 weeks old, male / female, purchased from CLEA Japan, SPF) were subjected to a 7-day quarantine and acclimation period, and it was confirmed that all mice were in good health and had not lost weight before being used in the study. The mice were reared in a 12-hour lighting environment with a temperature of 20-26°C and humidity of 30-70%, and were allowed to freely consume food and water.

[0067] Mouse colon cancer cell line (CT26) 1×10 6 The mice were then injected subcutaneously with 80μL of high-concentration Matrigel. From the 21st day after subcutaneous injection or when tumor formation was confirmed by palpation, whichever came first, tyrosine kinase inhibitors (TKI), immune checkpoint inhibitors (ICI), and Clostridium butyricum MIYAIRI 588 (FERM BP-2789) were administered to the following 5 groups: TKI group, TKI+CBM group, ICI group, TKI+ICI group, and TKI+ICI+CBM group (n=7 (TKI group, TKI+CBM group) and n=6 (ICI group, TKI+ICI group, TKI+ICI+CBM group)). In the control group (n=7), TKI, ICI, and CBM588 were not administered.

[0068] The mice were euthanized as a humane endpoint 28 days after the start of administration, or when the tumor mass reached 10% of the body weight or the tumor diameter reached 10 mm. 3 Mice exceeding this number were excluded.

[0069] Details of the drugs used and their dosage are shown below. TKI: Regorafenib (#R0142, Tokyo Chemical Industry Co., Ltd.) 30 μg / g, 100 μL (maximum amount 1000 μL) orally administered once a day for 9 days. ICI: Anti-CD279 antibody (J43; BE-00332, Iwai Chemicals Co., Ltd.) 5 μg / g, 100 μL (maximum amount 1600 μL) administered intraperitoneally every 3 days (continue until the end of the observation period) CBM: Miya BM powder (Miyarisan Pharmaceutical Co., Ltd.) 1 x 10 8 cfu / 100μL, 100μL administered orally three times a week.

[0070] On days 4, 8, 11, 15, and 18 after the start of administration, the tumor minor axis (mm) and tumor major axis (mm) were measured, and the tumor volume was calculated using the following formula: Tumor volume (mm 3 ) = 1 / 2 × tumor minor axis × tumor minor axis × tumor major axis.

[0071] The results are shown in Figures 1 and 2.

[0072] As shown in Figures 1 and 2, it can be seen that the combination of a tyrosine kinase inhibitor and CBM588 can suppress an increase in tumor volume.

[0073] In addition, tumor volume was 1000 mm 3 The Kaplan-Meier curve with events is shown in Figure 3. As shown in Figure 3, the log-rank test shows that the TKI+CBM group showed a significantly higher progression-free rate than the control group and the TKI group.

Claims

1. A combination drug for treating colorectal cancer, comprising a multikinase inhibitor and Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof.

2. The combination drug for treating colon cancer according to claim 1, wherein the Clostridium butyricum miyairi 588 is a live bacterium.

3. The combination pharmaceutical for treating colorectal cancer according to claim 1, wherein the multikinase inhibitor is at least one selected from the group consisting of axitinib, cabozantinib, sunitinib, sorafenib, pazopanib, regorafenib, and lenvatinib.

4. The combination drug for treating colorectal cancer described in claim 1, wherein the multikinase inhibitor is regorafenib.

5. The combination pharmaceutical for treating colorectal cancer according to claim 1 , further comprising an immune checkpoint inhibitor.

6. The combination drug for treating colorectal cancer according to claim 5, wherein the immune checkpoint inhibitor is at least one selected from the group consisting of a PD-1 inhibitor, a PD-L1 inhibitor, and a CTLA-4 inhibitor.

7. The combination drug for treating colorectal cancer according to claim 6, wherein the immune checkpoint inhibitor is at least one selected from the group consisting of nivolumab, ipilimumab, pembrolizumab, semipilimab, durvalumab, daclizumab, avelumab, and atezolizumab.

8. A pharmaceutical composition for use in treating colorectal cancer, comprising a combination of a multikinase inhibitor and Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof.

9. A pharmaceutical composition for treating colorectal cancer, comprising Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof, administered in combination with a multikinase inhibitor.

10. A pharmaceutical composition for treating colon cancer, comprising a multikinase inhibitor administered in combination with Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof.

11. A kit for treating colon cancer, comprising a multikinase inhibitor and Clostridium butyricum MIYAIRI 588 (FERM BP-2789) or a culture thereof.

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