Composition, screening method, and method for producing composition

Abstract

Provided are: a composition for treating cancer, the composition containing 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof; a composition for treating cancer, the composition containing a substance that promotes CX3CL1 gene expression; a composition for promoting CX3CL1 gene expression, the composition containing 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof; a screening method for selecting a substance that promotes CX3CL1 gene expression using CX3CL1 gene expression promotion as an index; or a method for producing a composition, the method including the screening method.

Description

Composition, screening method and method for producing composition

[0001] The present invention relates to compositions, screening methods and methods for producing compositions.

[0002] Cancer is the leading cause of death in Japan today, making its treatment extremely important. The main methods of cancer treatment include surgery, radiation therapy, and chemotherapy. Chemotherapy mainly includes chemotherapy using cytotoxic anticancer drugs and molecularly targeted drugs, hormone therapy using antihormonal drugs, and cancer immunotherapy using immune checkpoint inhibitors. However, due to individual differences in drug sensitivity and the development of drug resistance, new therapeutic drugs for cancer chemotherapy are constantly being sought. Effective therapeutic drugs for kidney cancer, in particular, are scarce, and their development is highly sought after.

[0003] Patent document 1 describes a pharmaceutical composition for treating abnormal cell growth in a mammal, comprising a specific imidazole derivative, or a pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof, and a pharmaceutically acceptable carrier. Patent document 2 describes specific pyridine, pyridazine, and triazine derivatives for use in cancer treatment.

[0004] On the other hand, 3-(p-hydroxyphenyl)propionic acid (also known as desaminotyrosine, 3-(4-hydroxyphenyl)propionic acid, phloretic acid, etc.) is a metabolite of flavonoids contained in food by intestinal bacteria. Non-Patent Document 1 describes that desaminotyrosine has the effect of protecting the host from influenza infection. Patent Document 3 describes a pharmaceutical composition for preventing, treating, or alleviating itching associated with skin, mucous membranes, or systemic diseases or disorders, comprising an effective amount of a phenylbutyric acid derivative and a pharmaceutically acceptable carrier, salt, or solvent thereof. Patent Document 4 describes that 3-(4-hydroxyphenyl)propionic acid (phloretic acid) does not exhibit cell killing activity against triple-negative breast cancer cell lines in a specific in vitro bioassay system.

[0005] Patent Publication No. 2008-515872, Patent Publication No. 2018-500351, Patent Publication No. 2009-51777, Patent Publication No. 2022-547721

[0006] Ashley L Steed et al. The microbial metabolite desaminotyrosine protects from influenza through type I interferon. Science. 2017 Aug 4; 357(6350): 498-502. doi: 10.1126 / science.aam5336

[0007] The present invention aims to provide a new use of 3-hydroxyphenylpropionic acid or a derivative thereof, or a salt thereof, and a new composition for treating cancer, which comprises a substance that promotes CX3CL1 gene expression. The present invention also aims to provide a new method for screening for a substance that promotes CX3CL1 gene expression, and a method for producing a new composition, which includes the screening method.

[0008] In a particular embodiment, the present invention aims to provide new compositions for the treatment of renal cancer (and further, renal cell carcinoma).

[0009] The present inventors have discovered that 3-hydroxyphenylpropionic acid, a derivative thereof, or a salt thereof exhibits anticancer activity, leading to the completion of the present invention. Furthermore, the present inventors have discovered that 3-hydroxyphenylpropionic acid, a derivative thereof, or a salt thereof promotes the expression of the CX3CL1 gene.

[0010] That is, the present invention relates to the following: [1] A composition for treating cancer, comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof. [2] The composition according to [1], comprising, as the 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, a compound represented by the following formula (1) or formula (2) or a pharmacologically acceptable salt of the compound: In formula (1), R Ｘeach independently represents a hydrogen atom or a substituent; L １ represents a single bond, —O—, or —C(═O)—, and Ar １ represents an aromatic ring structure, and R １ each independently represents a substituent, n1 represents an integer of 0 to 5, and when n1 is 2 or more, two or more R １ may be bonded to form a ring structure. Ｘ each independently represents a hydrogen atom or a substituent; L ２ represents a single bond, —O—, or —C(═O)—, and Ar ２ represents an aromatic ring structure, and R ２ each independently represents a substituent, n2 represents an integer of 0 to 5, and when n2 is 2 or more, two or more R ２ may be bonded to form a ring structure. [3] The composition according to [1] or [2], wherein the cancer treatment is mediated by promoting lymphocyte infiltration into cancer tissue. [4] The composition according to any one of [1] to [3], wherein the cancer treatment is mediated by promoting CX3CL1 gene expression in cells. [5] The composition according to any one of [1] to [4], wherein the cancer treatment is mediated by promoting CX3CL1 gene expression in cancer cells. [6] The composition according to any one of [1] to [5], wherein the cancer is renal cancer (further, renal cell carcinoma). [7] A composition for treating cancer, comprising a substance that promotes CX3CL1 gene expression. [8] The composition according to [7], wherein the active ingredient is a compound represented by the following formula (1) or formula (2) or a pharmacologically acceptable salt of the compound: In formula (1), R Ｘ each independently represents a hydrogen atom or a substituent; L １ represents a single bond, —O—, or —C(═O)—, and Ar １ represents an aromatic ring structure, and R １ each independently represents a substituent, n1 represents an integer of 0 to 5, and when n1 is 2 or more, two or more R １ may be bonded to form a ring structure. Ｘ each independently represents a hydrogen atom or a substituent; L ２ represents a single bond, —O—, or —C(═O)—, and Ar２ represents an aromatic ring structure, and R ２ each independently represents a substituent, n2 represents an integer of 0 to 5, and when n2 is 2 or more, two or more R ２ may be bonded to form a ring structure. [9] The composition according to [7] or [8], wherein the subject in which CX3CL1 gene expression is promoted is a kidney cell.

[10] A composition for promoting CX3CL1 gene expression, comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof.

[11] The composition according to

[10] , wherein the 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, comprises a compound represented by the following formula (1) or formula (2) or a pharmacologically acceptable salt of the compound: In formula (1), R Ｘ each independently represents a hydrogen atom or a substituent; L １ represents a single bond, —O—, or —C(═O)—, and Ar １ represents an aromatic ring structure, and R １ each independently represents a substituent, n1 represents an integer of 0 to 5, and when n1 is 2 or more, two or more R １ may be bonded to form a ring structure. Ｘ each independently represents a hydrogen atom or a substituent; L ２ represents a single bond, —O—, or —C(═O)—, and Ar ２ represents an aromatic ring structure, and R ２ each independently represents a substituent, n2 represents an integer of 0 to 5, and when n2 is 2 or more, two or more R ２may be bonded to form a ring structure.

[12] The composition according to

[10] or

[11] , which promotes CX3CL1 gene expression in cancer cells.

[13] The composition according to

[12] , wherein the cancer cells are renal cancer cells (or even cancerous cells of the renal parenchyma).

[14] The composition according to

[10] or

[11] , which promotes CX3CL1 gene expression in normal cells.

[15] The composition according to

[14] , wherein the normal cells are renal cells.

[16] A method for promoting CX3CL1 gene expression, comprising adding 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof to target cells.

[17] A screening method for selecting a substance that promotes CX3CL1 gene expression using promotion of CX3CL1 gene expression as an index.

[18] A method for producing a composition, comprising the screening method according to

[17] .

[0011] The present invention also relates to the following: <1> A method for treating cancer, comprising administering 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof to a subject. <2> A pharmaceutical composition comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, for use in the method for treating cancer. <3> Use of 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, in the manufacture of a composition for treating cancer. <2> The method, pharmaceutical composition, or use according to <1>, wherein the 3-hydroxyphenylpropionic acid or a derivative thereof, or the pharmacologically acceptable salt thereof, comprises a compound represented by the following formula (1) or formula (2), or a pharmacologically acceptable salt of the compound: In formula (1), R Ｘ each independently represents a hydrogen atom or a substituent; L １ represents a single bond, —O—, or —C(═O)—, and Ar １ represents an aromatic ring structure, and R １ each independently represents a substituent, n1 represents an integer of 0 to 5, and when n1 is 2 or more, two or more R １ may be bonded to form a ring structure. Ｘeach independently represents a hydrogen atom or a substituent; L ２ represents a single bond, —O—, or —C(═O)—, and Ar ２ represents an aromatic ring structure, and R ２ each independently represents a substituent, n2 represents an integer of 0 to 5, and when n2 is 2 or more, two or more R ２ may be bonded to form a ring structure. <3> The method, pharmaceutical composition, or use according to <1> or <2>, wherein the cancer treatment is mediated by promoting lymphocyte infiltration into cancer tissue. <4> The method, pharmaceutical composition, or use according to any one of <1> to <3>, wherein the cancer treatment is mediated by promoting CX3CL1 gene expression in cells. <5> The method, pharmaceutical composition, or use according to any one of <1> to <4>, wherein the cancer treatment is mediated by promoting CX3CL1 gene expression in cancer cells. <6> The method, pharmaceutical composition, or use according to any one of <1> to <5>, wherein the cancer is renal cancer (further, renal cell carcinoma). <7> A method for treating cancer, comprising administering to a subject a substance that promotes CX3CL1 gene expression. A pharmaceutical composition comprising a substance that promotes CX3CL1 gene expression for use in the method for treating cancer. Use of a substance that promotes CX3CL1 gene expression in the manufacture of a composition for treating cancer. <8> The method, pharmaceutical composition, or use according to <7>, comprising, as an active ingredient, a compound represented by the following formula (1) or formula (2) or a pharmacologically acceptable salt of the compound: In formula (1), R Ｘ each independently represents a hydrogen atom or a substituent; L １ represents a single bond, —O—, or —C(═O)—, and Ar １ represents an aromatic ring structure, and R １ each independently represents a substituent, n1 represents an integer of 0 to 5, and when n1 is 2 or more, two or more R １ may be bonded to form a ring structure. Ｘ each independently represents a hydrogen atom or a substituent; L ２ represents a single bond, —O—, or —C(═O)—, and Ar ２ represents an aromatic ring structure, and R ２each independently represents a substituent, n2 represents an integer of 0 to 5, and when n2 is 2 or more, two or more R ２ may be bonded to form a ring structure. <9> The method, pharmaceutical composition, or use according to <7> or <8>, wherein the subject in which CX3CL1 gene expression is promoted is a kidney cell. <10> A method for promoting CX3CL1 gene expression, comprising administering 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof to a subject. A pharmaceutical composition comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, for use in the method for promoting CX3CL1 gene expression. Use of 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, in the manufacture of a composition for promoting CX3CL1 gene expression. <11> The method, pharmaceutical composition, or use according to <10>, wherein the 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, comprises a compound represented by the following formula (1) or formula (2) or a pharmacologically acceptable salt of the compound: In formula (1), R Ｘ each independently represents a hydrogen atom or a substituent; L １ represents a single bond, —O—, or —C(═O)—, and Ar １ represents an aromatic ring structure, and R １ each independently represents a substituent, n1 represents an integer of 0 to 5, and when n1 is 2 or more, two or more R １ may be bonded to form a ring structure. Ｘ each independently represents a hydrogen atom or a substituent; L ２ represents a single bond, —O—, or —C(═O)—, and Ar ２ represents an aromatic ring structure, and R ２ each independently represents a substituent, n2 represents an integer of 0 to 5, and when n2 is 2 or more, two or more R ２may be bonded to form a ring structure. <12> The method, pharmaceutical composition, or use according to <10> or <11>, which promotes CX3CL1 gene expression in cancer cells. <13> The method, pharmaceutical composition, or use according to <12>, wherein the cancer cells are renal cancer cells (or even cancerous renal parenchymal cells). <14> The method, pharmaceutical composition, or use according to <10> or <11>, which promotes CX3CL1 gene expression in normal cells. <15> The method, pharmaceutical composition, or use according to <14>, wherein the normal cells are renal cells. <16> A method for treating cancer, which comprises administering 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof. <17> The method for treating cancer according to <16>, wherein the 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, comprises a compound represented by the following formula (1) or formula (2), or a pharmacologically acceptable salt of the compound: In formula (1), R Ｘ each independently represents a hydrogen atom or a substituent; L １ represents a single bond, —O—, or —C(═O)—, and Ar １ represents an aromatic ring structure, and R １ each independently represents a substituent, n1 represents an integer of 0 to 5, and when n1 is 2 or more, two or more R １ may be bonded to form a ring structure. Ｘ each independently represents a hydrogen atom or a substituent; L ２ represents a single bond, —O—, or —C(═O)—, and Ar ２ represents an aromatic ring structure, and R ２ each independently represents a substituent, n2 represents an integer of 0 to 5, and when n2 is 2 or more, two or more R ２ may be bonded to form a ring structure. <18> A method for treating cancer, comprising promoting expression of the CX3CL1 gene. <19> The method for treating cancer according to any one of <16> to <18>, which is a method for treating renal cancer.

[0012] In some embodiments, the present invention provides a composition for the treatment of cancer, comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof.

[0013] In some embodiments, the present invention provides a composition for promoting CX3CL1 gene expression, comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof. In some embodiments, the present invention provides a new composition for treating cancer, comprising a substance that promotes CX3CL1 gene expression. In some embodiments, the present invention provides a new method for screening for a substance that promotes CX3CL1 gene expression, and a method for producing a new composition, including the screening method.

[0014] Figure 1 shows the results of measuring the proportion of tumor tissue in the entire mouse kidney two weeks after tumor inoculation. HPPA (3-(p-hydroxyphenyl)propionic acid)-treated group vs. control (vehicle)-treated group. Figure 2 shows the results of measuring the proportion of (A) CD4+ T cells, (B) CD8+ T cells, or (C) NK cells relative to total immune cells (CD45+) in the kidney of mice two weeks after tumor inoculation. HPPA (p-HPPA)-treated group vs. control-treated group. Figure 3 shows the results of quantification of CX3CL1 gene expression in renal cancer Renca cells. HPPA (p-HPPA)-treated group vs. control group. Figure 4 shows the results of quantification of CX3CL1 gene expression in normal mouse kidney cells. HPPA (p-HPPA)-treated group vs. control group. Figure 5 shows the results of quantification of CX3CL1 gene expression in renal cancer Renca cells. Compounds A to L-treated group vs. control group. Figure 6 shows the results of quantification of CX3CL1 gene expression in renal cancer Renca cells. HPPA (o-HPPA, m-HPPA or p-HPPA) addition group vs. Control group.

[0015] The present invention will be described in detail below. The features of the present invention described below can be combined in any combination.

[0016] In some embodiments, the present invention relates to a composition comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof. Hereinafter, unless otherwise specified, the term "composition of the present invention" refers to a composition comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, regardless of its intended use. In some embodiments, the composition of the present invention is a pharmaceutical composition. In other embodiments, the composition of the present invention is a food composition. The composition of the present invention may be a liquid, a solid, or a mixture of a liquid and a solid.

[0017] A composition according to a first aspect of the present invention (hereinafter simply referred to as the "first composition") is a composition for treating cancer, comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof. A composition according to a second aspect of the present invention (hereinafter simply referred to as the "second composition") is a composition for treating cancer, comprising a substance that promotes CX3CL1 gene expression. A composition according to a third aspect of the present invention (hereinafter simply referred to as the "third composition") is a composition for promoting CX3CL1 gene expression, comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof. Hereinafter, any of the first, second, and third compositions will be referred to simply as the "composition of the present invention."

[0018] [Active ingredient] The first and third compositions of the present invention contain, as an active ingredient, 3-hydroxyphenylpropionic acid (sometimes referred to as 3-hydroxyphenylpropanoic acid, hydroxyhydrocinnamic acid, hydroxybenzenepropanoic acid, hydroxybenzenepropionic acid, hydrocoumaric acid, etc.; sometimes abbreviated as HPPA, etc.) having the structure shown below, or a derivative thereof, or a pharmacologically acceptable salt thereof. The second composition of the present invention preferably contains 3-hydroxyphenylpropionic acid having the structure shown below, or a derivative thereof, or a pharmacologically acceptable salt thereof.

[0019] (wherein the —OH of the phenyl group may be bonded to any position on the ring.)

[0020] In some embodiments, the 3-hydroxyphenylpropionic acid is 3-(p-hydroxyphenyl)propionic acid, which has been shown to have anti-cancer activity or CX3CL1 gene expression-promoting activity.

[0021] Derivatives of 3-hydroxyphenylpropionic acid include, for example, 3-phenylpropionic acid or 2-phenylpropionic acid, which are unsubstituted or have one or more substituents. The substituents may be bonded to each other to form a ring structure. The ring structure formed may be an aromatic ring structure or an aliphatic ring structure. Also, -C(=O)- or -O- may be inserted into 3-phenylpropionic acid or 2-phenylpropionic acid. When such insertion is present, it is particularly preferable that the benzene ring in 3-phenylpropionic acid or the benzene ring in 2-phenylpropionic acid is bonded to the alkylene group via -C(=O)- or -O-. These derivatives are also expected to have anticancer activity or CX3CL1 gene expression promoting activity. The substituents are not particularly limited, and examples thereof include C 1-3 Alkyl group, hydroxy group, phenyl group, carboxyl group, C 1-3 Alkoxyphenyl group (including o-, m-, and p-configurations), hydroxyphenyl group (including o-, m-, and p-configurations), C 1-3 Dialkylphenyl group (each C 1-3Each alkyl group may be bonded to any carbon atom of the phenyl group. ), hydroxyisobutylphenyl group (including o-, m-, and p-configurations. The hydroxy group may be bonded to any carbon atom of the isobutyl group.), halogen atoms (F, Cl, Br, etc.), and amino groups may be selected from the group consisting of phenyl, hydroxyisobutyl ... Specific examples of derivatives of 3-hydroxyphenylpropionic acid include 2-phenylpropionic acid, 2-methoxyphenylpropionic acid, 3-methoxyphenylpropionic acid (e.g., 3-(4-methoxyphenyl)propionic acid), 3-dimethylphenylpropionic acid, 2-ethoxy-3-hydroxyphenylpropionic acid, 3,3-diphenylpropionic acid, 2-methyl-2-phenylpropionic acid, 2-hydroxyphenylsuccinic acid, 3-hydroxyphenylbutyric acid, 3-hydroxyphenyl-3-hydroxypropionic acid, 3-hydroxyphenyl-2-hydroxypropionic acid, and 3-hydroxyphenyl-2-phenylpropionic acid. Examples of such propionic acids include 3-hydroxyphenyl-2-methylpropionic acid, 1-hydroxyibuprofen, 2-hydroxyibuprofen, 3-(p-tolyl)propionic acid, 3-(4-bromophenyl)propionic acid, 3-(4-chlorophenyl)propionic acid, 3-(3,4-methylenedioxyphenyl)propionic acid, 3-(p-methoxybenzoyl)propionic acid, 3-(4-fluorophenoxy)propionic acid, 3-(p-chlorophenoxy)propionic acid, 3-(4-aminophenyl)propionic acid, (2S)-2-(6-methoxy-2-naphthyl)propionic acid, and 3-(4-phenylbenzoyl)propionic acid.Among these, 3-(p-tolyl)propionic acid, 3-(4-bromophenyl)propionic acid, 3-(4-chlorophenyl)propionic acid, 3-(3,4-methylenedioxyphenyl)propionic acid, 3-phenylpropionic acid, 3-(p-methoxybenzoyl)propionic acid, 3-(4-fluorophenoxy)propionic acid, 3-(p-chlorophenoxy)propionic acid, 3-(4-methoxyphenyl)propionic acid, 3-(4-aminophenyl)propionic acid, (2S)-2-(6-methoxy-2-naphthyl)propionic acid, or 3-(4-phenylbenzoyl)propionic acid is preferred, (p-Tolyl)propionic acid, 3-(4-bromophenyl)propionic acid, 3-(4-chlorophenyl)propionic acid, 3-phenylpropionic acid, 3-(4-fluorophenoxy)propionic acid, 3-(p-chlorophenoxy)propionic acid, 3-(4-aminophenyl)propionic acid, or 3-(4-phenylbenzoyl)propionic acid is more preferred, and 3-(p-tolyl)propionic acid, 3-(4-chlorophenyl)propionic acid, 3-(4-fluorophenoxy)propionic acid, 3-(4-aminophenyl)propionic acid, or 3-(4-phenylbenzoyl)propionic acid is even more preferred. Furthermore, by bonding the substituents together, the benzene ring structure in 3-phenylpropionic acid or 2-phenylpropionic acid may form a naphthalene ring structure, a 1,2-methylenedioxybenzene ring structure, or the like.

[0022] The "pharmacologically acceptable salt" of 3-hydroxyphenylpropionic acid may be an acidic or basic salt. Examples of basic salts include, but are not limited to, alkali metal salts such as sodium and potassium, alkaline earth metal salts such as calcium and magnesium, ammonium salts, and salts of nitrogen-containing organic bases such as trimethylamine, triethylamine, tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, diethylamine, dicyclohexylamine, procaine, dibenzylamine, N-benzyl-β-phenethylamine, 1-ephenamine, and N,N'-dibenzylethylenediamine. Acid salts include, but are not limited to, mineral acid salts such as hydrochloride, hydrobromide, nitric acid, and sulfuric acid; organic carboxylic acid salts 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 sulfonic acid salts such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, mesitylenesulfonic acid, and naphthalenesulfonic acid. In some embodiments, the pharmacologically acceptable salt of 3-hydroxyphenylpropionic acid is an alkali metal salt. In other embodiments, the pharmacologically acceptable salt of 3-hydroxyphenylpropionic acid is a sodium salt.

[0023] 3-Hydroxyphenylpropionic acid or its derivatives or pharmacologically acceptable salts thereof may be obtained by any method, for example, commercially available products (e.g., manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). Alternatively, they may be synthesized by methods known to those skilled in the art. For example, in the case of 3-(p-hydroxyphenyl)propionic acid, its synthesis may involve hydrogenation of the 2-propenoic acid side chain of p-coumaric acid or hydrolysis of phloretin by phloretin hydrolase. Alternatively, 3-(p-hydroxyphenyl)propionic acid may be produced as a bacterial metabolite of flavonoids or via an unknown synthetic pathway. Examples of bacteria known to produce 3-(p-hydroxyphenyl)propionic acid through flavonoid metabolism include Clostridium orbiscindens (Non-Patent Document 1). Alternatively, 3-(p-hydroxyphenyl)propionic acid may be contained as a component of a bacterial fermentation product.

[0024] The content of 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof in the composition of the present invention is not particularly limited and can be set to an amount appropriate for the intended use, and may be, for example, 0.0001% (w / v) to 99.9999% (w / v).

[0025] The composition of the present invention preferably contains, as the 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, a compound represented by the following formula (1) or formula (2) or a pharmacologically acceptable salt of the compound: In formula (1), R Ｘ each independently represents a hydrogen atom or a substituent; L １ represents a single bond, —O—, or —C(═O)—, and Ar １ represents an aromatic ring structure, and R １ each independently represents a substituent, n1 represents an integer of 0 to 5, and when n1 is 2 or more, two or more R １ may be bonded to form a ring structure. Ｘ each independently represents a hydrogen atom or a substituent; L ２ represents a single bond, —O—, or —C(═O)—, and Ar２ represents an aromatic ring structure, and R ２ each independently represents a substituent, n2 represents an integer of 0 to 5, and when n2 is 2 or more, two or more R ２ may be bonded to form a ring structure.

[0026] In formula (1), R Ｘ are each independently a hydrogen atom, C １ ~C ３ Alkyl group, C １ ~C ３ In formula (1), L is preferably an alkoxy group, a hydroxy group, an amino group, a phenyl group, or a halogen atom (preferably F, Cl, or Br), more preferably a hydrogen atom, F, Cl, or Br, and even more preferably a hydrogen atom. １ represents a single bond, —O—, or —C(═O)—, and a single bond is preferred. １ represents an aromatic ring structure, preferably an aromatic hydrocarbon ring structure, more preferably a benzene ring structure or a naphthalene ring structure, and even more preferably a benzene ring structure. １ each independently represents a substituent, C １ ~C ３ Alkyl group, C １ ~C ３ An alkoxy group, a hydroxy group, an amino group, a phenyl group, or a halogen atom is preferred, and C １ ~C ３ An alkyl group, a hydroxy group, an amino group, a phenyl group, or a halogen atom is more preferred, and a methyl group, a hydroxy group, an amino group, a phenyl group, or a halogen atom is even more preferred. １ ~C ３ The alkyl group is preferably a methyl group. The halogen atom is preferably F, Cl, or Br, and more preferably F or Cl. In formula (1), n1 represents an integer of 0 to 5, preferably an integer of 1 to 3, and more preferably 1 or 2. An embodiment in which n1 is 1 is also one of the preferred embodiments of the present invention. When n1 is 2 or more, two or more R １ is bonded to Ar １Examples of the ring structure formed by containing R include a 1,2-methylenedioxybenzene ring structure. Among these, n1 is preferably 0 or n1 is 1 and R １ is C １ ~C ３ Alkyl group, C １ ~C ３ an alkoxy group, a hydroxy group, an amino group, a phenyl group, or a halogen atom, or n1 is 2 and two R １ is bonded to Ar １ and preferably comprises n1 to form a 1,2-methylenedioxybenzene ring structure, and n1 is 0 or n1 is 1 and R １ is C １ ~C ３ More preferably, n1 is an alkyl group, a hydroxy group, an amino group, a phenyl group, or a halogen atom, and n1 is 1 and R １ is C １ ~C ３ More preferably, it is an alkyl group, a hydroxy group, an amino group, a phenyl group, or a halogen atom. １ ~C ３ The alkyl group is preferably a methyl group. The halogen atom is preferably F, Cl or Br, more preferably F or Cl.

[0027] In formula (2), R Ｘ are each independently a hydrogen atom, C １ ~C ３ Alkyl group, C １ ~C ３ In formula (2), L is preferably an alkoxy group, a hydroxy group, an amino group, a phenyl group, or a halogen atom (preferably F, Cl, or Br), more preferably a hydrogen atom, F, Cl, or Br, and even more preferably a hydrogen atom. ２ represents a single bond, —O—, or —C(═O)—, and a single bond is preferable. ２ represents an aromatic ring structure, preferably an aromatic hydrocarbon ring structure, more preferably a benzene ring structure or a naphthalene ring structure, and even more preferably a naphthalene ring structure. ２ each independently represents a substituent, C １~C ３ Alkyl group, C １ ~C ３ An alkoxy group, a hydroxy group, an amino group, a phenyl group, or a halogen atom is preferred, and C １ ~C ３ An alkyl group, a hydroxy group, an amino group, a phenyl group, or a halogen atom is more preferred, and a methoxy group is even more preferred. In formula (2), n2 represents an integer of 0 to 5, preferably an integer of 1 to 3, and more preferably 1 or 2. An embodiment in which n2 is 1 is also one of the preferred embodiments of the present invention. When n2 is 2 or more, two or more R ２ is bonded to Ar ２ Examples of the ring structure formed by containing the above include a 1,2-methylenedioxybenzene ring structure.

[0028] The second composition of the present invention also contains a substance that promotes CX3CL1 gene expression. Examples of such substances include the aforementioned 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof. The substance that promotes CX3CL1 gene expression may also be selected by the screening method of the present invention described below. The first and third compositions of the present invention may each further contain a substance that promotes CX3CL1 gene expression selected by the screening method of the present invention described below.

[0029] [Uses] (Treatment of Cancer) In some embodiments, the composition of the present invention is a composition for treating cancer. In the present invention, "treatment" includes prevention or treatment. Furthermore, "prevention or treatment" of cancer includes shrinkage of cancer, reduction of cancer cells, inhibition of cancer growth, inhibition of cancer cell proliferation, inhibition of cancer recurrence, inhibition of cancer metastasis to lymph nodes and other tissues, inhibition of cancer incidence, improvement of cancer treatment efficacy, improvement of tumor markers, etc. Furthermore, "improvement" includes not only improvement of a disease, symptom, condition, etc., but also maintenance without worsening.

[0030] The size and metastasis of cancer can be confirmed by methods known to those skilled in the art, such as X-rays, computed tomography (CT), magnetic resonance imaging (MRI), ultrasound, and positron emission tomography (PET-CT). In animals such as mice, the size of cancer can be determined by, for example, collecting tumor tissue and measuring its size. The number of cancer cells can be determined, for example, by observing blood, cancer tissue, body fluids, etc. collected from a subject under an optical microscope. Tumor markers can be measured in body fluids such as blood and urine collected from a subject using any method appropriate for each tumor marker. For example, SCC (Squamous Cell Carcinoma), a tumor marker for squamous cell carcinoma, can be measured in serum collected from a subject using CLIA (chemiluminescence immunoassay). Each cancer-associated symptom can be evaluated using methods known to those skilled in the art. For example, cancer-associated pain can be evaluated using an appropriate scale, such as the Numerical Rating Scale (NRS), Visual Analogue Scale (VAS), or Verbal Rating Scale (VRS).

[0031] The type of cancer is not particularly limited, and examples include solid cancers such as colorectal cancer, lung cancer, gastric cancer, esophageal cancer, breast cancer, prostate cancer, pancreatic cancer, cervical cancer, uterine cancer, liver cancer, kidney cancer, ovarian cancer, head and neck cancer, bladder cancer, biliary tract cancer, adrenal cancer, thyroid cancer, and malignant melanoma; sarcomas such as osteosarcoma, chondrosarcoma, liposarcoma, undifferentiated pleomorphic sarcoma, myxofibrosarcoma, and leiomyosarcoma; and hematopoietic cancers such as leukemia, malignant lymphoma, and multiple myeloma. In some embodiments, the cancer does not include triple-negative breast cancer (breast cancer that is estrogen receptor-, progesterone receptor-, and human epidermal growth factor receptor type 2-negative). In some embodiments, the cancer is a CX3CL1-expressing cancer. In some embodiments, the cancer is renal cancer (or even renal cell carcinoma). Because renal cells have a limited number of chemokines that induce leukocyte migration, the expression level of CX3CL1, a type of chemokine, is believed to be important in cancer treatment, particularly in renal cancer, as described below. Therefore, the composition of the present invention is particularly advantageous for the treatment of renal cancer or when added to renal cells (including both normal renal cells and renal cells in pathological conditions, such as renal cancer cells). For these reasons, the composition is believed to be particularly effective in cancers whose primary site is the kidney (i.e., renal cancer, particularly renal cell carcinoma), and is particularly effective against cancers whose primary site is the kidney and located in the kidney as a primary focus. The inventors have observed that oral administration of an antibiotic cocktail to tumor model mice resulted in significant tumor progression in tumor models in which renal cancer cells were implanted in the renal cortex compared to tumor models in which renal cancer cells were implanted subcutaneously. Oral administration of the antibiotic cocktail is believed to reduce HPPA produced by intestinal bacteria in the mouse intestine. Therefore, it is speculated that "3-hydroxyphenylpropionic acid or its derivatives or pharmacologically acceptable salts thereof" such as HPPA may be responsible for tumor suppression, particularly in the kidney.In addition, there have been reports of using the expression level of CX3CL1 as a prognostic marker for renal cancer (BMC Cancer. 2022 Nov 17;22(1):1184.), but there have been no reports of using CX3CL1 expression as a target molecule in cancer treatment.

[0032] In some embodiments, cancer treatment involves immune activation. In the body, cancer cells are recognized as non-self and are primarily killed by lymphocytes. In some embodiments, cancer treatment with the compositions of the present invention is mediated by promoting lymphocyte infiltration into cancer tissue (the effect of immune cell infiltration into cancer tissue, the effect of enhancing the migration of immune cells into cancer tissue). Promoting lymphocyte infiltration into cancer tissue promotes cancer cell damage. In some embodiments, the lymphocytes are one or more types selected from T cells and NK cells. In another embodiment, the lymphocytes are one or more types selected from CD4+ T cells and CD8+ T cells, or both. Lymphocyte infiltration into cancer tissue can be confirmed by any method known to those skilled in the art. For example, the method may involve staining lymphocytes in a portion of cancer tissue collected from a subject with fluorescently labeled antibodies against lymphocyte-specific cell surface markers and observing the lymphocytes under a fluorescence microscope.

[0033] In some embodiments, cancer treatment with the compositions of the present invention is mediated by promoting CX3CL1 gene expression in cells (somatic cells, particularly renal cells). In some embodiments, the treatment is mediated by promoting CX3CL1 gene expression in cancer cells. CX3CL1 (also known as CX3C chemokine or fractalkine) is a type of chemokine known to correlate with T cell chemotaxis (Patent Documents 5 and 6). In other words, the compositions of the present invention promote CX3CL1 gene expression in cancer cells, thereby enhancing the ability of T cells to migrate to cancer tissues and promoting T cell-mediated damage to cancer cells. CX3CL1 gene expression includes the transcription of the CX3CL1 gene into mRNA and the production of CX3CL1 protein from the CX3CL1 gene. Promotion of CX3CL1 gene expression includes increasing the amount of CX3CL1 gene mRNA expression (e.g., promoting mRNA transcription) and increasing the amount of CX3CL1 protein synthesis (e.g., promoting mRNA translation). The promotion of CX3CL1 gene expression can be evaluated by methods known to those skilled in the art, such as extracting mRNA from cells, quantifying the expression level of the CX3CL1 gene by real-time qPCR, and comparing it with any control. Alternatively, the method may involve quantifying the CX3CL1 protein produced by gene expression by techniques known to those skilled in the art, such as methods using absorbance or gel electrophoresis, and comparing it with any control. Furthermore, in some embodiments, the promotion of CX3CL1 gene expression by the compositions of the present invention is promotion of CX3CL1 gene expression in normal cells. Promotion of CX3CL1 gene expression in normal cells enhances the migration ability of the above-mentioned T cells to tissues, which may be effective in preventing cancer and treating other diseases.

[0034] (Promotion of CX3CL1 Gene Expression) In some embodiments, the composition of the present invention is a composition for promoting CX3CL1 gene expression. In some embodiments, the composition of the present invention is a composition for promoting CX3CL1 gene expression in cancer cells. In some embodiments, the composition of the present invention is a composition for promoting CX3CL1 gene expression in one or more cancer cells selected from the group consisting of renal cancer cells, malignant melanoma cells, non-small cell lung cancer cells, colon cancer cells, gastric cancer cells, bladder cancer cells, ovarian cancer cells, breast cancer cells, head and neck cancer cells, pancreatic cancer cells, sarcoma cells, esophageal cancer cells, small cell lung cancer cells, multiple myeloma cells, non-Hodgkin's lymphoma cells, and endometrial cancer cells. In some embodiments, the composition of the present invention is a composition for promoting CX3CL1 gene expression in renal cancer cells (and further, renal cell carcinoma, which is a malignant tumor formed by cancerous transformation of renal parenchymal cells). Furthermore, in some embodiments, the target cells in which CX3CL1 gene expression is promoted may be any of ectodermal cells, mesodermal cells, and endodermal cells, including, but not limited to, epithelial cells, endothelial cells, and blood cells. In some embodiments, renal cells are preferred. The target cells may be mutated due to canceration or artificial mutation. Furthermore, in some embodiments, the compositions of the present invention are compositions for promoting CX3CL1 gene expression in normal cells. In some embodiments, the compositions of the present invention are compositions for promoting CX3CL1 gene expression in cells constituting body tissues, such as cells in one or more cells selected from the kidney, adrenal gland, lung, bronchi, intestine, stomach, pancreas, liver, spleen, gallbladder, esophagus, appendix, bladder, ovary, testis, breast, oral cavity, head and neck, eyeball, blood cell, uterus, heart, brain, blood vessels, skin, bone, and other mucous membranes. In some embodiments, the compositions of the present invention are compositions for promoting CX3CL1 gene expression in renal cells.

[0035] In some embodiments, the compositions of the present invention for promoting CX3CL1 gene expression are suitable for administration to subjects in need of or desirably promoting CX3CL1 gene expression, including, for example, those suffering from cancer, those in need of or desirably preventing cancer recurrence or metastasis after treatment (including surgical therapy, radiation therapy, chemotherapy, etc.), and those at high risk of developing cancer (including smokers, those with a history of infection with Helicobacter pylori, hepatitis B virus, hepatitis C virus, human papillomavirus, etc., and those with a history of hormone therapy, etc.). The cancer may be solid cancer such as colon cancer, lung cancer, stomach cancer, esophageal cancer, breast cancer, prostate cancer, pancreatic cancer, cervical cancer, uterine cancer, liver cancer, kidney cancer, ovarian cancer, head and neck cancer, bladder cancer, biliary tract cancer, adrenal cancer, thyroid cancer, or malignant melanoma; sarcoma such as osteosarcoma, chondrosarcoma, liposarcoma, undifferentiated pleomorphic sarcoma, myxofibrosarcoma, or leiomyosarcoma; hematopoietic cancer such as leukemia, malignant lymphoma, or multiple myeloma; or in some embodiments, renal cancer (and further, renal cell carcinoma). Alternatively, the composition of the present invention for promoting CX3CL1 expression may be administered to subjects suffering from an infectious disease (including bacterial infection, viral infection, fungal infection, etc.) or at high risk of contracting an infectious disease (e.g., elderly people; subjects receiving treatment including administration of anticancer drugs, immunosuppressants, steroids, etc.; subjects with underlying diseases such as chronic kidney disease, chronic obstructive pulmonary disease, obesity, heart disease, diabetes, asthma, cerebrovascular disease, acquired immune deficiency syndrome, dementia, liver disease, etc.).

[0036] In some embodiments, the composition of the present invention for promoting CX3CL1 gene expression may not be intended to be administered to a subject (i.e., a living body). Non-limiting examples of compositions not intended to be administered to a living body include those intended to be used in in vitro experiments.

[0037] (Subject of administration, route of administration, dosage form, dosage amount, other components, etc.) When the composition of the present invention is administered to a subject, the subject may be, for example, a mammal, preferably a human or mouse, more preferably a human. The age (or age in months or weeks) and sex of the mammal are not particularly limited. When the subject is a human, it includes elderly people, adults, children, infants, and babies.

[0038] When the composition of the present invention is administered to a subject, the route of administration is not particularly limited as long as it is suitable for the purpose, and may be, for example, oral administration, enteral administration (including gastrostomy, enterostomy, nasogastric tube administration, etc.), intravenous administration, subcutaneous administration, intramuscular administration, rectal administration, intradermal administration, intrathecal administration, intraarterial administration, intranasal administration, oral mucosal administration, etc. In some embodiments, the composition of the present invention is for enteral administration (including oral administration, gastrostomy, enterostomy, nasogastric tube administration, etc.). In other embodiments, the composition of the present invention is for oral administration.

[0039] The compositions of the present invention can be formulated into dosage forms suitable for the administration route. For example, non-limiting examples of dosage forms suitable for oral administration include tablets, capsules, powders, fine granules, granules, liquids, and syrups. Non-limiting examples of dosage forms suitable for administration routes other than oral administration include injections, infusions, suppositories, inhalants, nasal drops, transdermal absorbents, ointments, creams, and patches. Furthermore, HPPA is known to be a bacterial metabolite of flavonoids and other substances. To enhance HPPA production in the body, CX3CL1 gene expression may be promoted by administering a live HPPA-producing bacterial agent.

[0040] When the composition of the present invention is administered to a subject, the dosage, frequency, and number of administrations can be determined as desired depending on the purpose of treatment, severity of the disease, the subject's age, weight, sex, body surface area, general condition, excretory function, treatment history, interactions with other drugs, the success of treatment, the presence or absence of side effects, etc. Non-limiting examples of dosages include a range of 0.01 μg / kg to 10 g / kg of active ingredient per day for an adult. Non-limiting examples of dosage frequencies and number of administrations include once a month, once a week, once a day, twice a day, three times a day, etc.

[0041] When the composition of the present invention is administered to a subject, it may be administered in combination with other active ingredients. Combination with other active ingredients means that the composition of the present invention is administered simultaneously with or sequentially to the other active ingredients, or at any time within a therapeutic regimen using the other active ingredients. The other active ingredients may be, but are not limited to, anticancer agents, antiemetics that can be used in chemotherapy regimens, antiallergic agents, steroids, or antibiotics (antibacterial agents, antiviral agents, antifungal agents), etc. Anticancer agents include, for example, cytotoxic anticancer agents such as DNA alkylating agents, platinum compounds, anticancer antibiotics, antimetabolites, DNA topoisomerase inhibitors, and mitotic inhibitors; molecular targeted agents such as anti-CD20 antibodies, anti-HER2 antibodies, anti-VEGF antibodies, tyrosine kinase inhibitors, kinase inhibitors, proteasome inhibitors, and mTOR inhibitors; antihormonal agents such as antiestrogens and antiandrogens; immune checkpoint inhibitors such as anti-PD-1 antibodies, anti-PD-L1 antibodies, and anti-CTLA-4 antibodies; and cell therapies such as CAR-T cells. The composition of the present invention may contain other active ingredients.

[0042] The composition of the present invention may contain any additives as long as the effects of the present invention can be obtained. Examples of additives include excipients, disintegrants, binders, lubricants, coating agents, pigments, bases, solubilizers, isotonicity agents, pH adjusters, preservatives, thickeners, antioxidants, and flavoring agents. Non-limiting examples of additives that can be used in preparing dosage forms for oral administration include excipients such as glucose, lactose, D-mannitol, starch, and crystalline cellulose; disintegrants or disintegration aids such as carboxymethylcellulose, starch, and carboxymethylcellulose calcium; binders such as hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, and gelatin; lubricants such as magnesium stearate and talc; coating agents such as hydroxypropylmethylcellulose, sucrose, polyethylene glycol, and titanium oxide; and bases such as petrolatum, liquid paraffin, polyethylene glycol, gelatin, kaolin, glycerin, purified water, and hard fat. Non-limiting examples of additives that can be used to prepare dosage forms for injection or infusion include solubilizers or solubilizing aids that can constitute aqueous or ready-to-use injections, such as distilled water for injection, physiological saline, propylene glycol, surfactants, etc.; isotonicity agents such as glucose, sodium chloride, D-mannitol, glycerin, etc.; and pH adjusters such as inorganic acids, organic acids, inorganic bases, organic bases, etc.

[0043] The composition of the present invention can be prepared by any method known to those skilled in the art. For example, the composition of the present invention can be prepared by dissolving 3-hydroxyphenylpropionic acid or a pharmacologically acceptable salt thereof, and optionally other components or additives, in a suitable solvent and mixing them. The suitable solvent may be, for example, an aqueous sodium chloride solution such as physiological saline.

[0044] (Method for Promoting CX3CL1 Gene Expression) The method for promoting CX3CL1 gene expression of the present invention is a method for promoting CX3CL1 gene expression, which comprises adding 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof to a subject cell.

[0045] Preferred embodiments of 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof are the same as the preferred embodiments of 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof in the composition of the present invention described above.

[0046] The target cells may be ectodermal, mesodermal, or endodermal cells, including, but not limited to, epithelial cells, endothelial cells, and blood cells. In some embodiments, renal cells are preferred. The target cells may be mutated by carcinogenesis or artificial mutation. Examples of cancer cells used in the method for promoting CX3CL1 gene expression of the present invention include renal cancer cells, malignant melanoma cells, non-small cell lung cancer cells, colon cancer cells, gastric cancer cells, bladder cancer cells, ovarian cancer cells, breast cancer cells, head and neck cancer cells, pancreatic cancer cells, sarcoma cells, esophageal cancer cells, small cell lung cancer cells, multiple myeloma cells, non-Hodgkin's lymphoma cells, and endometrial cancer cells. The origin of the target cells is not limited, and includes mammals such as humans, dogs, cats, rodents (e.g., rats and mice), lagomorphs, non-human primates, and ungulates (e.g., horses, cows, pigs, sheep, and goats), with humans being preferred.

[0047] The method for adding the composition to the target cells is not particularly limited, and known methods can be used in vitro, in situ, ex vivo, in vivo, etc. For example, methods commonly used to promote gene expression under these environments can be used without particular limitation. Alternatively, the composition may be added to the target cells by administration to the subject in the same manner as the administration of the composition of the present invention to the subject described above.

[0048] (Screening Method) The screening method of the present invention is a screening method for selecting a substance that promotes CX3CL1 gene expression using promotion of CX3CL1 gene expression as an index.

[0049] The screening method of the present invention can be carried out, for example, using cultured cells. One example preferably includes adding a candidate substance to the cultured cells and, after adding the candidate substance, confirming the expression of the CX3CL1 gene in the cultured cells.

[0050] Cultured cells may be ectodermal, mesodermal, or endodermal cells, including, but not limited to, epithelial cells, endothelial cells, and blood cells. In some embodiments, renal cells are preferred. The target cells may be mutated by carcinogenesis or artificial mutation. Examples of cancer cells used in the screening method of the present invention include renal cancer cells, malignant melanoma cells, non-small cell lung cancer cells, colon cancer cells, gastric cancer cells, bladder cancer cells, ovarian cancer cells, breast cancer cells, head and neck cancer cells, pancreatic cancer cells, sarcoma cells, esophageal cancer cells, small cell lung cancer cells, multiple myeloma cells, non-Hodgkin's lymphoma cells, and endometrial cancer cells. Screening using cancer cells may also be preferable for examining their usefulness in cancer treatment. Furthermore, the origin of the cultured cells is not limited, and examples include mammals such as humans, dogs, cats, rodents (rats, mice, etc.), lagomorphs, non-human primates, and ungulates (horses, cows, pigs, sheep, goats, etc.), with humans being preferred.

[0051] The candidate substance is not particularly limited and may be any compound, one of which is the aforementioned 3-hydroxyphenylpropionic acid or its derivatives, or pharmacologically acceptable salts thereof. The present invention also encompasses a stepwise screening method, such as using a substance containing multiple substances, such as a food-derived substance, as a substance, and identifying which of the components contained therein is an active ingredient after confirming CX3CL1 gene expression.

[0052] The method for adding a candidate substance to cultured cells is not particularly limited, and known methods can be used. For example, the candidate substance may be added to the culture medium of the cultured cells. The timing, amount, and duration of addition may be appropriately determined by referring to known methods for confirming the promotion of gene expression.

[0053] As a method for confirming CX3CL1 gene expression in cultured cells, known methods can be used without particular limitation, including Northern blotting, real-time PCR, microarray, RNA sequencing, etc. Conditions and the like for these methods can be determined based on known protocols.

[0054] Here, for example, a test is performed on a group administered with a candidate substance and a group not administered with the candidate substance, with each group being n=4 or more (preferably n=10 or more, more preferably n=25 or more), and if a significant increase in the amount of gene expression promotion is observed, the substance can be determined to be a substance that promotes CX3CL1 gene expression. A conventional statistical method, such as a t-test, can be used to test whether or not there is a significant increase.

[0055] (Method for Producing a Composition) The method for producing a composition of the present invention includes the screening method of the present invention. For example, a method includes mixing a substance determined to be a substance that promotes CX3CL1 gene expression by the screening method of the present invention with another substance. Examples of other substances include other active ingredients in the composition of the present invention described above and components described as optional additives. Furthermore, a food product containing a substance determined to be a substance that promotes CX3CL1 gene expression by the screening method of the present invention can also be used as a composition of the present invention. The content of the substance that promotes CX3CL1 gene expression in the composition is not particularly limited and can be selected according to the intended use, and may be, for example, 0.0001% (w / v) to 99.9999% (w / v).

[0056] The present invention will be described below with reference to examples, but the present invention should not be construed as being limited thereto.

[0057] Example 1: Tumor Model and Method for Evaluation of Antitumor Effect. BALB / c mice (male, 8-9 weeks old, Jackson Laboratory Japan, p-HPPA (3-(p-hydroxyphenyl)propionic acid) and control: 11 mice per group) were deeply anesthetized with the triple-analgesic mixture. A 1-cm incision was made in the left flank, and Renca cells derived from mouse renal carcinoma were transplanted into the nearby left kidney (1 × 10^5 cells / mouse) using a 28G needle. The incision was then sutured. For the p-HPPA group, HPPA (p-HPPA: Fujifilm Wako Pure Chemical Industries) was administered ad libitum in the form of sodium salt at 40 mM in drinking water from 1 week before transplantation until euthanasia. For the control group, sodium chloride at the same concentration was administered ad libitum in drinking water. Water intake was approximately 4 mL / mouse per day. Two weeks after transplantation, the mice were euthanized, and the left kidney where the cancerous tissue had formed was removed and bisected with a scalpel. The cross-sections were then pathologically examined, and the proportion of tumor tissue in the total kidney area was measured using a microscope (Keyence, BZ-X810). All mice were administered an antibiotic mixture (ampicillin 1 g / L, streptomycin 5 g / L, colistin 1 g / L) in their drinking water ad libitum from two weeks before transplantation until the time of euthanasia to eliminate the influence of intestinal bacteria.

[0058] In the above test, two weeks after tumor transplantation, tumor growth in the kidneys of mice was significantly suppressed in the p-HPPA-treated group ("HPPA" in Figure 1) compared with the control group ("Control" in Figure 1) (Figure 1). Testing was performed using a nonparametric two-sample Mann-Whitney U test (two-tailed distribution).

[0059] Example 2: Evaluation of the tissue infiltration effect of CD4+ T cells, CD8+ T cells, and NK cells. A comparative study was conducted between mice treated with p-HPPA and a control group (five mice per group) as in Example 1. The left kidneys containing cancerous tissue were physically sheared with a scalpel and enzymatically dispersed with collagenase to prepare cell suspensions. After necessary fluorescently labeled antibody staining, tumor-bearing kidney-infiltrating lymphocytes were analyzed using a flow cytometer. The proportion of CD4+ T cells (CD45+TCRβ+CD4+), CD8+ T cells (CD45+TCRβ+CD8a+), and NK cells (CD45+TCRβ-NK1.1+) relative to total immune cells (CD45+) was measured.

[0060] As a result, CD4+ T cells (Fig. 2A) and CD8+ T cells (Fig. 2B), which are important cells for antitumor immunity, were significantly increased in the p-HPPA-treated group ("HPPA" in Fig. 2) compared with the control group ("Control" in Fig. 2), and NK cells (Fig. 2C) also tended to increase, demonstrating that p-HPPA promotes the infiltration of immune cells such as CD4+ T cells and CD8+ T cells into major tissues (enhancing their migration ability). The analysis was performed using a Student's t test (two-tailed distribution) on two samples with equal variances.

[0061] Example 3: CX3CL1 Gene Expression. Subcultured adherent Renca cells were seeded onto 48-well plates and cultured until semi-confluent (approximately 70-80% of the adhesive surface of the culture vessel was occupied by cultured cells). After 24 hours of stimulation with 250 μL / well of RPMI-1640 medium (Gibco) containing 500 μM of the test substance (p-HPPA), the cells were harvested. mRNA was extracted from the cells using the Maxwell® RSC simplyRNA Cells Kit (Promega). Real-time qPCR (Quant Studio 3, Applied Biosystems) was performed to calculate relative quantification values, calculated from the number of PCR cycles required to amplify a given amount of mouse CX3CL1 gene expression, normalized by the expression level of the housekeeping gene β-actin. A control group was also tested in the same manner, but without any medium added, and relative quantification values ​​were calculated. Primers for the βactin and CX3CL1 genes were purchased from Takara Bio Inc. (NM_007393.5 and NM_009142.3, respectively).

[0062] As a result, the gene expression of CX3CL1, a chemokine important in attracting immune cells, was significantly increased in the p-HPPA-treated group ("HPPA" in Figure 3) compared to the control group ("Control" in Figure 3) (Figure 3). This indicates that p-HPPA has the effect of increasing (enhancing) CX3CL1 gene expression. The test was performed using a Student's t test (two-tailed distribution) on two samples with equal variances.

[0063] Example 4: CX3CL1 Gene Expression (Normal Cells) Six BALB / c mice (male, 8 weeks old, Jackson Laboratory Japan) were euthanized, and their kidneys were harvested. They were then physically sheared with a scalpel and enzymatically digested with collagenase to prepare cell suspensions. These cells were seeded into 96-well plates (1 x 10^6 cells per well) and stimulated with 200 μL / well of RPMI-1640 medium (Gibco) containing 500 μM p-HPPA for 24 hours. mRNA was extracted from the cells (Maxwell® RSC simplyRNA Cells Kit, Promega). Real-time qPCR (Quant Studio 3, Applied Biosystems) was used to quantify the expression level of the mouse CX3CL1 gene relative to the number of PCR cycles required for amplification of a given amount of nucleic acid (corrected for the expression level of the housekeeping gene Gapdh). A similar experiment was also performed as a control, with the medium being void of any additives, and relative quantification values ​​were calculated. Primers for the Gapdh and CX3CL1 genes were purchased from Takara Bio Inc. (NM_008084.3 and NM009142.3, respectively).

[0064] In normal renal cells, the gene expression of CX3CL1, a chemokine important for attracting immune cells, was significantly increased in the p-HPPA-treated group (p-HPPA in Figure 4) compared to the control group (Control in Figure 4) (Figure 4). The test was performed using a Student's t test (two-tailed distribution) on two samples with equal variances.

[0065] Example 5: CX3CL1 Gene Expression (Compounds Other Than p-HPPA) Subcultured adherent Renca cells were seeded onto a 96-well plate and cultured until semi-confluent (approximately 70-80% of the adhesive surface of the culture vessel was occupied by cultured cells). Test substances (A-L, as described below) were dissolved in RPMI-1640 medium (Gibco) at 200 μL / well for 24 hours. After stimulation, mRNA was extracted from the cells (Maxwell® RSC simplyRNA Cells Kit, Promega). Expression levels of the mouse CX3CL1 gene were quantified relative to the number of PCR cycles required for amplification of a given amount of nucleic acid (corrected for the expression level of the housekeeping gene, β-actin) using real-time qPCR (Quant Studio 3, Applied Biosystems). The stimulation concentration was all 500 μM. Details of the test substances A-L listed in Figure 5 are as follows. The following compounds A-L are compounds that correspond to derivatives of 3-hydroxyphenylpropionic acid in the present invention. For the control group, the same experiment was performed without adding anything to the medium, and relative quantitative values ​​were calculated. Primers for the β-actin and CX3CL1 genes were purchased from Takara Bio Inc. (NM_007393.5 and NM_009142.3, respectively). A: 3-(p-tolyl)propionic acid B: 3-(4-bromophenyl)propionic acid C: 3-(4-chlorophenyl)propionic acid D: 3-(3,4-methylenedioxyphenyl)propionic acid E: 3-phenylpropionic acid F: 3-(p-methoxybenzoyl)propionic acid G: 3-(4-fluorophenoxy)propionic acid H: 3-(p-chlorophenoxy)propionic acid I: 3-(4-methoxyphenyl)propionic acid J: 3-(4-aminophenyl)propionic acid K: (2S)-2-(6-methoxy-2-naphthyl)propionic acid L: 3-(4-phenylbenzoyl)propionic acid

[0066] In the compound A to L-treated groups ("A" to "L" in Figure 5), gene expression of CX3CL1, a chemokine important in attracting immune cells, was significantly increased compared to the control group ("Control" in Figure 5) (Figure 5). In Figure 5, * indicates a P value of less than 0.05, ** indicates a P value of less than 0.01, *** indicates a P value of less than 0.001, and **** indicates a P value of less than 0.0001. The significance level was determined by one-way analysis of variance using Dunnett's multiple comparisons with the control group.

[0067] Example 6: CX3CL1 Gene Expression (o-HPPA, m-HPPA) Subcultured adherent Renca cells were seeded onto 48-well plates and cultured until semi-confluent (approximately 70-80% of the adhesive surface of the culture vessel was occupied by cultured cells). After 24 hours of stimulation with test substances (p-HPPA, m-HPPA, o-HPPA) dissolved in 250 μL / well of RPMI-1640 medium (Gibco), mRNA was extracted from the cells (Maxwell® RSC simplyRNA Cells Kit, Promega). Expression levels of the mouse CX3CL1 gene were quantified relative to the number of PCR cycles required for amplification of a given amount of nucleic acid (corrected for the expression level of the housekeeping gene β-actin) using real-time qPCR (Quant Studio 3, Applied Biosystems). The stimulation concentration was 500 μM for all experiments. For the control, a similar experiment was performed without adding any medium, and relative quantification values ​​were calculated. Primers for the βactin and CX3CL1 genes were purchased from Takara Bio Inc. (NM_007393.5 and NM_009142.3, respectively). Details of the test substances, p-HPPA, m-HPPA, and o-HPPA, are as follows: p-HPPA: 3-(p-hydroxyphenyl)propionic acid, m-HPPA: 3-(m-hydroxyphenyl)propionic acid, o-HPPA: 3-(o-hydroxyphenyl)propionic acid.

[0068] In the groups treated with m-HPPA ("m-HPPA" in Figure 6) or o-HPPA ("o-HPPA" in Figure 6), no significant increase in gene expression of CX3CL1, a chemokine important for attracting immune cells, was observed compared to the control group ("Control" in Figure 6) (Figure 6). In the group treated with p-HPPA ("p-HPPA" in Figure 6), a significant increase in gene expression of CX3CL1 was observed compared to the control group ("Control" in Figure 6). Testing was performed using one-way analysis of variance using Dunnett's multiple comparisons with the control group.

[0069] These results suggest that p-HPPA and its derivative compounds have the ability to induce CX3CL1 production, and that the induction of CX3CL1 production enhances antitumor renal immunity and suppresses tumor growth.

[0070] (References cited in the detailed description of the invention) Patent Document 5: JP 2021-118684 Patent Document 6: JP 2019-60883

Claims

1. A composition for the treatment of cancer comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof.

2. The composition according to claim 1, comprising, as the 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, a compound represented by the following formula (1) or formula (2) or a pharmacologically acceptable salt of the compound: In formula (1), R Ｘ each independently represents a hydrogen atom or a substituent; L １ represents a single bond, -O- or -C(=O)-; Ar １ represents an aromatic ring structure; R １ each independently represents a substituent, n1 represents an integer of 0 to 5, and when n1 is 2 or more, two or more R １ may be bonded to form a ring structure. Ｘ each independently represents a hydrogen atom or a substituent; L ２ represents a single bond, -O- or -C(=O)-; Ar ２ represents an aromatic ring structure; R ２ each independently represents a substituent, n2 represents an integer of 0 to 5, and when n2 is 2 or more, two or more R ２ may be linked to form a ring structure.

3. The composition according to claim 1, wherein the treatment of cancer is via promoting lymphocyte infiltration into cancer tissue.

4. The composition according to claim 1, wherein the treatment of cancer is via promoting CX3CL1 gene expression in cells.

5. The composition according to claim 1, wherein the treatment of cancer is via promoting CX3CL1 gene expression in cancer cells.

6. The composition according to any one of claims 1 to 5, wherein the cancer is renal cancer.

7. A composition for treating cancer, comprising a substance that promotes CX3CL1 gene expression.

8. The composition according to claim 7, comprising as an active ingredient a compound represented by the following formula (1) or (2) or a pharmacologically acceptable salt of said compound: In formula (1), R Ｘ each independently represents a hydrogen atom or a substituent; L １ represents a single bond, -O- or -C(=O)-; Ar １ represents an aromatic ring structure; R １ each independently represents a substituent, n1 represents an integer of 0 to 5, and when n1 is 2 or more, two or more R １ may be bonded to form a ring structure. Ｘ each independently represents a hydrogen atom or a substituent; L ２ represents a single bond, -O- or -C(=O)-; Ar ２ represents an aromatic ring structure; R ２ each independently represents a substituent, n2 represents an integer of 0 to 5, and when n2 is 2 or more, two or more R ２ may be linked to form a ring structure.

9. The composition according to claim 7 or 8, wherein the subject in which CX3CL1 gene expression is promoted is a renal cell.

10. A composition for promoting CX3CL1 gene expression, comprising 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof.

11. The composition according to claim 10, comprising, as the 3-hydroxyphenylpropionic acid or a derivative thereof, or a pharmacologically acceptable salt thereof, a compound represented by the following formula (1) or formula (2) or a pharmacologically acceptable salt of the compound: In formula (1), R Ｘ each independently represents a hydrogen atom or a substituent; L １ represents a single bond, -O- or -C(=O)-; Ar １ represents an aromatic ring structure, R １ each independently represents a substituent, n1 represents an integer of 0 to 5, and when n1 is 2 or more, two or more R １ may be bonded to form a ring structure. Ｘ each independently represents a hydrogen atom or a substituent; L ２ represents a single bond, -O- or -C(=O)-; Ar ２ represents an aromatic ring structure; R ２ each independently represents a substituent, n2 represents an integer of 0 to 5, and when n2 is 2 or more, two or more R ２ may be linked to form a ring structure.

12. The composition described in claim 10, which promotes CX3CL1 gene expression in cancer cells.

13. The composition according to claim 12, wherein the cancer cells are renal cancer cells.

14. The composition described in claim 10, which promotes CX3CL1 gene expression in normal cells.

15. The composition of claim 14, wherein the normal cells are renal cells.

16. A screening method for selecting a substance that promotes CX3CL1 gene expression using promotion of CX3CL1 gene expression as an indicator.

17. A method for producing a composition comprising the screening method according to claim 16.

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