Gene expression inhibitor and gene expression inhibiting composition

Abstract

To provide a gene expression inhibitor that inhibits expression of any one gene selected from a group consisting of an Arid5a gene, a Rac2 gene, a Clic4 gene, a keratin gene family and an Eps15 gene.SOLUTION: The present invention provides a gene expression inhibitor that inhibits expression of any one gene selected from a group consisting of an Arid5a gene, a Rac2 gene, a Clic4 gene, a keratin gene family and an Eps15 gene, the gene expression inhibitor comprising at least one of a glycyrrhizinic acid and a tranexamic acid.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an expression inhibitor for any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene; a composition for inhibiting the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene; a composition for at least one of preventing and treating a disease caused by the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene; and an expression inhibitor for two or more genes selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene. [Background technology]

[0002] AT-rich interactive domain-containing protein 5A (hereinafter also referred to as "Arid5a") is a protein that binds to mRNA of Interleukin-6 (hereinafter also referred to as "IL-6"), an inflammatory cytokine. In the absence of Arid5a, IL-6 mRNA is quickly degraded by Regnase-1, a ribonuclease, even if it is produced. However, in the presence of Arid5a, Arid5a binds to the 3' Untranslated Region (UTR) of IL-6 mRNA, protecting IL-6 mRNA from degradation by Regnase-1, resulting in abnormal production of IL-6, which has been suggested to promote autoimmune diseases and aggravate inflammation (Non-Patent Document 1).

[0003] Rac Family Small GTPase 2 (hereinafter, also referred to as "Rac2") is a protein that interacts with nitric oxide synthase 2 (hereinafter, also referred to as "NOS2") and activates NOS2. NOS2 is known to be involved in the synthesis of nitric oxide, which aggravates inflammation and the like (Non-Patent Document 2).

[0004] Chloride intracellular channel 4 (hereafter referred to as Clic4) is a cytoplasmic protein that is involved in various actin-dependent processes, such as integrin trafficking, cell adhesion, and vacuole formation. In keratinocytes, increased expression of Clic4 is associated with increased Ca 2+ It is associated with differentiation by keratinocytes, DNA damage, and increased expression of p53 and tumor necrosis factor-α (TNF-α) (Non-Patent Documents 3, 4, 5). Clic4 is not only present in the cytoplasm but is also a component of the inner mitochondrial membrane, and maintenance of the Clic4 expression level is essential for the survival of keratinocytes (Non-Patent Document 4). In fact, it has been reported that even a slight increase in Clic4 expression significantly increases differentiation into keratinocytes (Non-Patent Document 6).

[0005] Keratin, a major component of the epithelial cytoskeleton, contributes to the structural stability of keratinocytes. More than 54 mammalian keratins have been identified to date, and they are broadly classified into acidic Type I keratin and neutral / basic Type II keratin based on pH (Non-Patent Documents 7, 8, 9). Keratin forms heterodimers consisting of Type I and Type II keratins, which further assemble to form intermediate filaments, a type of cytoskeletal fiber.

[0006] There are several types of keratin, including keratin 1, 5, 14, 16, and 17. Keratin 1 (hereinafter, also referred to as "Krt1") is a differentiation marker for keratinocytes (Non-Patent Documents 10 and 11). Krt1 is produced together with Keratin 10 (Non-Patent Document 12). Keratin 5 (hereinafter also referred to as "Krt5") has a molecular weight of 56 kDa and an isoelectric point of 7.4 (Non-Patent Document 7). As the primary keratin in basal cells, it is produced together with keratin 14 (hereinafter also referred to as "Krt14") (Non-Patent Document 13). Krt5 is produced in basal cells with high mitotic activity in stratified epithelium. Production of Krt5 correlates with mitotic activity of epithelial cells, but does not correlate with epithelial cell differentiation or epithelial stratification (Non-Patent Document 14). In humans, Krt14 has a molecular weight of 50 kDa and an isoelectric point of 5.3 (Non-Patent Documents 7 and 15). Krt14 expression correlates with the degree of mitotic activity and pluripotency of basal cells in stratified epithelium (Non-Patent Document 16). Keratin 16 (hereinafter also referred to as "Krt16") has a molecular weight of 46 kDa and an isoelectric point of 5.1 (Non-Patent Document 7). Krt16 is a marker for cells presumed to be in an intermediate state of cell differentiation between basal and suprabasal cells, and provides sufficient structural stability by providing the flexibility required for cell movement and mitosis (Non-Patent Document 17). Krt16 and its basic partner Keratin 6 are markers for the activated state of keratinocytes and contribute to migration, extracellular matrix production, and hyperplasia (Non-Patent Document 18). Keratin 17 (hereinafter also referred to as "Krt17") has a molecular weight of 48 kDa and an isoelectric point of 5.1 (Non-Patent Document 19). Krt17 is involved in wound healing and developmental processes (Non-Patent Document 19). Krt17 is expressed simultaneously with Krt6 and forms a heterodimer with Krt6, but it can also bind to Krt5 and Keratin 8 (Non-Patent Document 19).

[0007] Epidermal growth factor receptor pathway substrate clone 15 (hereinafter also referred to as "Eps15") is known to contribute to receptor-mediated endocytosis of epidermal growth factor (Non-Patent Document 23). [Prior art documents] [Non-patent literature]

[0008]

Linked Book1

Outdoor Tool2

Outdoor Tools3

Outdoor Tools 4

Direct Environment 5

[0009] As described above, Arid5a leads to aggravation of inflammation. Establishing a means for controlling the expression of a gene encoding Arid5a (hereinafter also referred to as "Arid5a gene") is highly meaningful in preventing and / or treating diseases caused by the expression of the Arid5a gene.

[0010] In addition, Rac2 causes inflammation in the kidney, etc. Establishing a means for controlling the expression of a gene encoding Rac2 (hereinafter also referred to as "Rac2 gene") is highly significant in preventing and / or treating diseases caused by the expression of the Rac2 gene.

[0011] There are diseases caused by the excessive production of Clic4 by keratinocytes. Establishing a means for controlling the expression of the gene encoding Clic4 (hereinafter also referred to as "Clic4 gene") is very meaningful in preventing and / or treating diseases caused by Clic4 gene expression.

[0012] When a wound is formed in the skin, keratinocytes on the basement membrane undergo a process called keratinocyte activation (Non-Patent Documents 20, 21). These keratinocytes do not follow the normal differentiation pathway, but are induced to contribute to wound repair, for example, by increasing the expression of Krt16 after injury. During the process of keratinocyte activation, when cells assume a migratory morphology, cell adhesion between keratinocytes is dramatically reduced, and the number of desmosome attachments between cells decreases (Non-Patent Document 22). It is presumed that the reduction in the number of desmosome attachments of keratinocytes on the basement membrane makes the epithelial tissue brittle and prone to peeling, which triggers the formation of ulcers. It is believed that suppressing the overexpression of a group of genes encoding keratin produced by keratinocytes on the basement membrane (hereinafter also referred to as the "keratin gene family") is very meaningful in preventing and / or treating diseases caused by the expression of the keratin gene family.

[0013] Eps15 contributes to the receptor-mediated endocytosis of epidermal growth factor. It is feared that endocytosis is the main component of biological reactions involving innate immunity. Establishing a means for suppressing the expression of the gene encoding Eps15 (hereinafter also referred to as the "Eps15 gene") is believed to be highly significant in the prevention and / or treatment of diseases caused by Eps15 gene expression.

[0014] Therefore, the present invention aims to provide an agent for inhibiting the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, as well as a composition for inhibiting the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene. [Means for solving the problem]

[0015] As a result of extensive research, the inventors have found that the administration or ingestion of an agent consisting of at least one of glycyrrhizinic acid and tranexamic acid, or a composition containing at least one of glycyrrhizinic acid and tranexamic acid as an active ingredient, can suppress the expression of the Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene, and have completed the present invention.

[0016] That is, the present invention is as follows. [1] An agent for inhibiting the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, comprising at least one of (A) glycyrrhizinic acid and (B) tranexamic acid. [2] A composition for inhibiting the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, comprising at least one of (A) glycyrrhizinic acid and (B) tranexamic acid as an active ingredient. [3] A composition for at least one of preventing and treating a disease caused by the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, comprising the gene expression inhibitor described in [1] above. [4] A composition for at least one of the prevention and treatment of a disease caused by the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene described in [3] above, wherein the disease caused by the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene is stomatitis. [5] An agent for inhibiting the expression of two or more genes selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, comprising at least one of (A) glycyrrhizinic acid and (B) tranexamic acid.

[0017] Further, one aspect of the present invention is as follows. A method for suppressing the expression of any one gene selected from the group consisting of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene, comprising administering or ingesting at least one of (A) glycyrrhizinic acid and (B) tranexamic acid to a subject. A method for suppressing the expression of two or more genes selected from the group consisting of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene, comprising administering or ingesting at least one of (A) glycyrrhizinic acid and (B) tranexamic acid to a subject. A method for suppressing the expression of any one gene selected from the group consisting of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene, comprising administering or ingesting at least one of (A) glycyrrhizinic acid and (B) tranexamic acid to a subject (excluding humans). A method for suppressing the expression of two or more genes selected from the group consisting of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene, comprising administering or having a subject (excluding human individuals) ingest at least one of (A) glycyrrhizinic acid and (B) tranexamic acid. Effect of the Invention

[0018] According to the present invention, the administration or ingestion of the agent or composition of the present invention can effectively suppress the expression of at least one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene. Furthermore, the administration or ingestion of the agent or composition of one aspect of the present invention can effectively prevent or treat one or more diseases selected from the group consisting of diseases caused by the expression of the Arid5a gene, diseases caused by the expression of the Rac2 gene, diseases caused by the expression of the Clic4 gene, diseases caused by the expression of the keratin gene family, and diseases caused by the expression of the Eps15 gene. [Brief description of the drawings]

[0019] [Figure 1] FIG. 1 is a graph showing the expression level ratio of Arid5a gene in Test Example 1. [Diagram 2] FIG. 2 is a graph showing the expression level ratio of Rac2 gene in Test Example 1. [Diagram 3] FIG. 3 is a graph showing the expression level ratio of Clic4 gene in Test Example 1. [Figure 4A] FIG. 4A is a graph showing the expression level ratio of Krt1 genes in Test Example 1. [Figure 4B] FIG. 4B is a graph showing the expression level ratio of Krt14 gene in Test Example 1. [Figure 4C] FIG. 4C is a graph showing the expression level ratio of Krt16 gene in Test Example 1. [Figure 4D] FIG. 4D is a graph showing the expression level ratio of Krt17 gene in Test Example 1. [Diagram 5] FIG. 5 is a graph showing the expression level ratio of Eps15 genes in Test Example 1. [Figure 6] FIG. 6 is a graph showing the expression level ratio of Pla2g2f genes in Test Example 1. [Figure 7] FIG. 7 is a graph showing the expression level of the Pla2g2f gene relative to the expression level of GAPDH in Test Example 2. [Figure 8] FIG. 8 is a graph showing the expression level of the Arid5a gene relative to the expression level of GAPDH in Test Example 2. [Figure 9] FIG. 9 is a graph showing the expression level of the Rac2 gene relative to the expression level of GAPDH in Test Example 2. [Figure 10] FIG. 10 is a graph showing the expression level of the Clic4 gene relative to the expression level of GAPDH in Test Example 2. [Figure 11A] FIG. 11A is a graph showing the expression level of the Krt1 gene relative to the expression level of GAPDH in Test Example 2. [Figure 11B] FIG. 11B is a graph showing the expression level of the Krt5 gene relative to the expression level of GAPDH in Test Example 2. [Figure 11C] FIG. 11C is a graph showing the expression level of the Krt14 gene relative to the expression level of GAPDH in Test Example 2. [Figure 11D] FIG. 11D is a graph showing the expression level of the Krt16 gene relative to the expression level of GAPDH in Test Example 2. [Figure 12] FIG. 12 is a graph showing the expression level of the Eps15 gene relative to the expression level of GAPDH in Test Example 2. [Figure 13] FIG. 13 is a graph showing the expression level of the Pla2g2f gene relative to the expression level of GAPDH in Test Example 3. [Figure 14] FIG. 14 is a graph showing the expression level of the Arid5a gene relative to the expression level of GAPDH in Test Example 3. [Figure 15]FIG. 15 is a graph showing the expression level of the Rac2 gene relative to the expression level of GAPDH in Test Example 3. [Figure 16] FIG. 16 is a graph showing the expression level of the Clic4 gene relative to the expression level of GAPDH in Test Example 3. [Figure 17A] FIG. 17A is a graph showing the expression level of the Krt1 gene relative to the expression level of GAPDH in Test Example 3. [Figure 17B] FIG. 17B is a graph showing the expression level of the Krt5 gene relative to the expression level of GAPDH in Test Example 3. [Figure 17C] FIG. 17C is a graph showing the expression level of the Krt14 gene relative to the expression level of GAPDH in Test Example 3. [Figure 17D] FIG. 17D is a graph showing the expression level of the Krt16 gene relative to the expression level of GAPDH in Test Example 3. [Figure 18] FIG. 18 is a graph showing the expression level of the Eps15 gene relative to the expression level of GAPDH in Test Example 3. [Figure 19A] FIG. 19A is a photograph of the right buccal mucosa of a hamster immediately after intradermal injection of an EGFP gene solution in Reference Example 1. [Figure 19B] FIG. 19B is a photograph of the hamster buccal mucosa emitting fluorescent wavelengths derived from EGFP protein in Reference Example 1. [Figure 20A] FIG. 20A is a photograph showing immunohistochemical staining performed on an untreated hamster in Reference Example 1. [Figure 20B] FIG. 20B is a photograph showing immunohistochemical staining of the gene transfer site in a hamster into which the EGFP gene was transferred in Reference Example 1. [Figure 21] FIG. 21 is a graph showing the degree of inflammatory edema (stomatitis) in hamsters in which the Arid5a gene or the Rac2 gene was knocked down and in control hamsters in Reference Example 2. [Figure 22]FIG. 22 is a graph showing the area of ​​inflammatory edema (stomatitis) in Pla2g2f gene knockdown hamsters and control hamsters in Reference Example 2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] The present invention will be described in detail below, but these are merely examples of preferred embodiments and the present invention is not limited to these contents. The "to" in a numerical range includes the numerical values ​​before and after it. For example, "0% by mass to 100% by mass" means a range that is equal to or greater than 0% by mass and equal to or less than 100% by mass.

[0021] [Gene expression inhibitors] The present invention relates to a gene expression inhibitor for any one selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, and a gene expression inhibitor for two or more selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, which comprises at least one of (A) glycyrrhizic acid (hereinafter also referred to as the (A) component) and (B) tranexamic acid (hereinafter also referred to as the (B) component). In the present specification, the present invention encompasses a gene expression inhibitor for any one selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, and a gene expression inhibitor for two or more selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene. As described later, the glycyrrhizic acid (A) contained in the agent of the present invention may be in any form, such as its original form, its salt form, or the form of a plant extract containing glycyrrhizic acid. Also, as described later, the tranexamic acid (B) contained in the agent of the present invention may be in any form, such as its original form, its salt form, or its derivative or its salt form.

[0022] In the present specification, the Arid5a gene includes the human-derived ARID5A gene (NCBI Gene accession number 10865) and a homologue in the human-derived ARID5A gene. When aligned with the base sequence of the human-derived ARID5A gene, the homologue in the human-derived ARID5A gene has a base sequence identity of preferably 70% or more, more preferably 80% or more, and even more preferably 90% or more. The homologue in the human-derived ARID5A gene may be a gene derived from a human or a gene derived from an animal other than a human. Examples of animals other than a human include hamsters, dogs, cats, and rabbits.

[0023] In this specification, the Rac2 gene includes human-derived RAC2 gene (NCBI Gene accession number 5880) and homologues in human-derived RAC2 gene. When aligned with the base sequence of the human-derived RAC2 gene, the homologues in human-derived RAC2 gene have a base sequence identity of preferably 70% or more, more preferably 80% or more, and even more preferably 90% or more. The homologues in human-derived RAC2 gene may be human-derived genes or may be genes derived from animals other than humans. Examples of animals other than humans include hamsters, dogs, cats, rabbits, etc.

[0024] In the present specification, the Clic4 gene is the human CLIC4 gene (NCBI Gene Accession No. 25932), and homologs in the human-derived CLIC4 gene. A homolog in the human-derived CLIC4 gene has a base sequence identity of preferably 70% or more, more preferably 80% or more, and even more preferably 90% or more when aligned with the base sequence of the human-derived CLIC4 gene. A homolog in the human-derived CLIC4 gene may be a gene derived from a human, or may be a gene derived from an animal other than a human. Examples of animals other than a human include hamsters, dogs, cats, and rabbits.

[0025] Examples of the keratin gene family include the gene encoding Krt1 (hereinafter also referred to as the "Krt1 gene"), the gene encoding Krt5 (hereinafter also referred to as the "Krt5 gene"), the gene encoding Krt14 (hereinafter also referred to as the "Krt14 gene"), the gene encoding Krt16 (hereinafter also referred to as the "Krt16 gene"), and the gene encoding Krt17 (hereinafter also referred to as the "Krt17 gene"). As used herein, the Krt1 gene includes the human KRT1 gene (NCBI Gene accession number 3848) and homologs of the human KRT1 gene. As used herein, the Krt5 gene includes the human KRT5 gene (NCBI Gene accession number 3852) and homologs of the human KRT5 gene. As used herein, the Krt14 gene includes the human KRT14 gene (NCBI Gene accession number 3861) and homologs of the human KRT14 gene. As used herein, the Krt16 gene includes the human KRT16 gene (NCBI Gene accession number 3868) and homologs of the human KRT16 gene. As used herein, the Krt17 gene includes the human KRT17 gene (NCBI Gene accession number 3872) and homologs of the human KRT17 gene. The homologues of the human KRT1 gene, the human KRT5 gene, the human KRT14 gene, the human KRT16 gene, and the human KRT17 gene are preferably 70% or more, more preferably 80% or more, and even more preferably 90% or more when aligned with the base sequences of the human KRT1 gene, the human KRT5 gene, the human KRT14 gene, the human KRT16 gene, and the human KRT17 gene. The homologues of the human KRT1 gene, the human KRT5 gene, the human KRT14 gene, the human KRT16 gene, and the human KRT17 gene may be human genes or may be genes from animals other than humans. Examples of animals other than humans include hamsters, dogs, cats, and rabbits.

[0026] In the present specification, the Eps15 gene is the human EPS15 gene (NCBI Gene accession number 2060) and homologs in the human-derived EPS15 gene. When aligned with the base sequence of the human-derived EPS15 gene, the homologs in the human-derived EPS15 gene have a base sequence identity of preferably 70% or more, more preferably 80% or more, and even more preferably 90% or more. The homologs in the human-derived EPS15 gene may be genes derived from humans or may be genes derived from animals other than humans. Examples of animals other than humans include hamsters, dogs, cats, and rabbits.

[0027] <(A) Glycyrrhizic acid> Glycyrrhizic acid (ingredient (A)) is a component contained in the roots and stems of licorice, a type of herbal medicine. The present inventors have unexpectedly discovered that administration or ingestion of ingredient (A) suppresses the expression of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene.

[0028] Component (A) may be used as it is, or may be used in the form of a salt. Examples of salts of glycyrrhizinic acid include potassium salts, sodium salts, and ammonium salts, and specific examples include monopotassium glycyrrhizinate, dipotassium glycyrrhizinate, monosodium glycyrrhizinate, disodium glycyrrhizinate, monoammonium glycyrrhizinate, and diammonium glycyrrhizinate. As described above, glycyrrhizinic acid is known to be contained in licorice extract, and may be used in the form of a plant extract containing glycyrrhizinic acid. The component (A) may be used alone or in combination of two or more types. Among these, from the viewpoints of increasing water solubility, facilitating formulation, or increasing affinity for the epithelium, it is preferable to use glycyrrhizinate, with potassium salts and ammonium salts of glycyrrhizinic acid being more preferable, and dipotassium glycyrrhizinate and monoammonium glycyrrhizinate being even more preferable.

[0029] <(B) Tranexamic acid> Tranexamic acid (ingredient (B)) is a type of amino acid, also known as trans-4-aminomethylcyclohexane-1-carboxylic acid. As with ingredient (A), the present inventors have unexpectedly found that administration or ingestion of ingredient (B) suppresses the expression of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene.

[0030] The component (B) may be used in its original form or in the form of a salt. Examples of salts of tranexamic acid include alkali metal salts such as sodium and potassium; alkaline earth metal salts such as calcium and magnesium; metal salts such as aluminum, iron, and zinc; basic amino acid salts such as lysine, arginine, histidine, and ornithine; and organic amine salts such as ammonium, monoethanolamine, diethanolamine, triethanolamine, and stearylamine. The component (B) may be used as a tranexamic acid derivative or a salt thereof, and specific examples thereof include ester derivatives such as tranexamic acid cetyl ester; and amide derivatives such as tranexamic acid methylamide.

[0031] The amount of administration or intake of the agent of the present invention is not particularly limited as long as it does not impair the effects of the present invention, and can be appropriately changed depending on various factors such as the age, condition, etc. of the subject to which it is applied. The amount of administration or intake preferable for achieving the desired effect may be appropriately determined depending on the application of the agent.

[0032] The agent of the present invention can be used as it is as a final product. It can also be used as an additive for food and drink, an additive for medicine, and an additive for quasi-drugs. This can impart various effects to food and drink, medicine, and quasi-drugs.

[0033] [Composition for inhibiting gene expression] The composition for inhibiting the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, and the composition for inhibiting the expression of two or more genes selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene of the present invention contain at least one of the above-mentioned components (A) and (B) as active ingredients, and may further contain the component (C) described below, but may further contain any component other than the above-mentioned components (A), (B), and (C) within a range that does not inhibit the effects of the present invention. In this specification, the composition of the present invention includes the composition for inhibiting the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, and the composition for inhibiting the expression of two or more genes selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene.

[0034] For example, at least one of the components (A) and (B) is preferably contained in the composition of the present invention in an amount of 0.001 to 1 w / v%. Here, the contents of the components (A) and (B) refer to the contents converted into glycyrrhizinic acid and tranexamic acid, respectively. That is, when the component (A) is blended in the form of a plant extract containing glycyrrhizinic acid or used in the form of a glycyrrhizinate, the amount of the component (A) is converted into glycyrrhizinic acid, and when the component (B) is used in the form of a tranexamate, a tranexamic acid derivative, or a salt thereof, the amount of the component (B) is contained in the above range in the amount converted into tranexamic acid. The above conversion can be carried out, for example, by analyzing a plant extract containing glycyrrhizinic acid or a glycyrrhizinate salt for component (A), and a tranexamate salt or a tranexamic acid derivative or a salt thereof for component (B) by a standard liquid chromatography method using a liquid chromatograph or high performance liquid chromatograph, and confirming the content of glycyrrhizinic acid for component (A) and the content of tranexamic acid for component (B).

[0035] When the content of at least one of the (A) and (B) components is 0.001 w / v% or more, the effect of the present invention is easily exhibited, and when it is 1 w / v% or less, the composition of the present invention has an excellent feeling of use as a preparation, such as no irritation, stickiness, and flavor if it is used in the oral cavity, and the preparation stability tends to be increased. The content of at least one of the (A) and (B) components is preferably 0.003 w / v% or more, more preferably 0.005 w / v% or more, more preferably 0.7 w / v% or less, even more preferably 0.31 w / v% or less, even more preferably 0.30 w / v% or less, particularly preferably 0.25 w / v% or less, and most preferably 0.2 w / v% or less.

[0036] The content of the component (A) in the composition of the present invention is preferably in the range of 0.001 to 0.31 w / v% in terms of glycyrrhizinic acid. When the content of the component (A) is 0.001 w / v% or more in terms of glycyrrhizinic acid, the desired effect of the present invention is easily obtained, and when the content is 0.31 w / v% or less, the composition of the present invention has an excellent feeling of use as a preparation, such as irritation, stickiness, and flavor if used in the oral cavity, and the stability of the preparation also tends to be improved. The content of the component (A) is more preferably 0.003 w / v% or more in terms of glycyrrhizinic acid, even more preferably 0.005 w / v% or more, even more preferably 0.006 w / v% or more, and particularly preferably 0.009 w / v% or more. Also, it is more preferably 0.30 w / v% or less, even more preferably 0.25 w / v% or less, and particularly preferably 0.2 w / v% or less.

[0037] The (B) component is preferably contained in the composition of the present invention in an amount of 0.001 to 1 w / v% calculated as tranexamic acid. When the (B) component content is 0.001 w / v% or more, the desired effect of the present invention is easily obtained, and when the (B) component content is 1 w / v% or less, the composition of the present invention has an excellent feeling of use as a preparation, such as no irritation or stickiness, and flavor if used in the oral cavity, and the preparation stability also tends to be increased. The (B) component content is more preferably 0.003 w / v% or more, even more preferably 0.005 w / v% or more, even more preferably 0.01 w / v% or more, particularly preferably 0.03 w / v% or more, and most preferably 0.05 w / v% or more. Furthermore, it is more preferably 0.7 w / v% or less, even more preferably 0.31 w / v% or less, even more preferably 0.30 w / v% or less, particularly preferably 0.25 w / v% or less, and most preferably 0.2 w / v% or less.

[0038] <(C) Quaternary ammonium salt> The composition of the present invention may contain a quaternary ammonium salt (hereinafter, also referred to as component (C)). Quaternary ammonium salts are ingredients known to have germicidal effects.

[0039] Examples of quaternary ammonium salts include cetylpyridinium chloride, benzethonium chloride, benzalkonium chloride, dequalinium chloride, alkyldimethylammonium chloride, alkyltrimethylammonium chloride, methylbenzethonium chloride, lauroylcholaminoformylmethylpyridinium chloride, etc. The component (C) may be used alone or in combination of two or more. Among these, cetylpyridinium chloride, benzethonium chloride, and benzalkonium chloride are preferred, and cetylpyridinium chloride is more preferred, since they are highly effective in suppressing the expression of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene.

[0040] The quaternary ammonium salt is preferably contained in the composition of the present invention in the range of 0.005 to 0.1 w / v%. When the content of the quaternary ammonium salt is 0.005 w / v% or more, the desired effect of the present invention tends to be easily obtained, and when it is 0.1 w / v% or less, the composition of the present invention has an excellent feeling of use as a preparation, such as irritation, stickiness, and flavor if used in the oral cavity, and the preparation stability also tends to be improved. The content of the quaternary ammonium salt is more preferably 0.01 w / v% or more, more preferably 0.02 w / v% or more, more preferably 0.08 w / v% or less, and even more preferably 0.07 w / v% or less.

[0041] The contents of (A) glycyrrhizinic acid, (B) tranexamic acid, and (C) quaternary ammonium salt in the composition of the present invention can be measured by a conventional liquid chromatography method using a liquid chromatograph or a high performance liquid chromatograph.

[0042] The agent or composition of the present invention preferably contains both component (A) and component (B), which can more effectively suppress the expression of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene. When the composition of the present invention contains both the (A) component and the (B) component, the total content of the (A) component and the (B) component is preferably 0.001 to 1.31 w / v%. The total content of the (A) component and the (B) component is more preferably 0.003 w / v% or more, even more preferably 0.005 w / v% or more, even more preferably 0.01 w / v% or more, particularly preferably 0.03 w / v% or more, and most preferably 0.05 w / v% or more. Also, the total content of the (A) component and the (B) component is more preferably 1.30 w / v% or less, even more preferably 1.25 w / v% or less, and particularly preferably 1.2 w / v% or less.

[0043] Furthermore, the composition of the present invention preferably contains all of the components (A), (B) and (C). By containing these three components, additive or synergistic enhancement of the effects can be obtained.

[0044] In the agent or composition of the present invention, the content ratio A / B of the component (A) to the component (B) is preferably in the range of 0.001 to 310. When the content ratio A / B is in the above range, the effects of the present invention can be more effectively exhibited. The content ratio A / B is more preferably 0.003 or more, even more preferably 0.005 or more, even more preferably 0.01 or more, particularly preferably 0.05 or more, most preferably 0.1 or more, more preferably 300 or less, even more preferably 200 or less, even more preferably 150 or less, especially preferably 110 or less, particularly preferably 100 or less, and most preferably 50 or less.

[0045] In the composition of the present invention, the content ratio A / C of the component (A) to the component (C) is preferably in the range of 0.01 to 60. When the content ratio A / C is in the above range, the effects of the present invention can be more effectively exhibited. The content ratio A / C is more preferably 0.03 or more, even more preferably 0.05 or more, even more preferably 0.08 or more, particularly preferably 0.1 or more, more preferably 50 or less, even more preferably 30 or less, even more preferably 20 or less, particularly preferably 10 or less.

[0046] In the composition of the present invention, the content ratio B / C of the component (B) to the component (C) is preferably in the range of 0.01 to 200. When the content ratio B / C is in the above range, the effects of the present invention can be more effectively exhibited. The content ratio B / C is more preferably 0.03 or more, even more preferably 0.05 or more, even more preferably 0.08 or more, particularly preferably 0.1 or more, more preferably 120 or less, even more preferably 60 or less, even more preferably 50 or less, even more preferably 30 or less, particularly preferably 20 or less, and most preferably 10 or less.

[0047] Optional components that may be contained in the composition of the present invention include, for example, solvents, sweeteners, bactericides, surfactants, anti-inflammatory agents, tartar prevention agents, pH adjusters, enzymes, ion sources, preservatives, fragrances, herbal medicines, pigments, moisturizing ingredients, cell activation ingredients, vitamins, amino acids, etc.

[0048] Examples of the solvent include water such as purified water and ionized water; physiological saline; lower monohydric alcohols such as ethanol; and polyhydric alcohols such as propylene glycol and polyethylene glycol.

[0049] Examples of sweeteners include stevioside, rebaudioside, xylitol, erythritol, maltitol, sorbitol, sodium saccharin, sucralose, trehalose, reduced palatinose, aspartame, and acesulfame potassium.

[0050] Examples of disinfectants include hinokitiol, triclosan, isopropylmethylphenol, thymol, alkyldiaminoethylglycine hydrochloride solution, chlorhexidine gluconate, chlorhexidine hydrochloride, and sodium chlorite.

[0051] Examples of the surfactant include nonionic surfactants, cationic surfactants, anionic surfactants, and amphoteric surfactants. Specifically, examples of the nonionic surfactants include glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, polyoxyethylene fatty acid esters, polyoxyethylene sorbitan fatty acid esters, fatty acid diethanolamides, fatty acid monoglycerides, polyoxyethylene alkyl ethers, polyoxyethylene hydrogenated castor oil, alkyl glucosides, and polyoxyethylene polyoxypropylene block copolymers. Examples of the cationic surfactants include polyoxyethylene alkylamines and fatty acid amides. Examples of the anionic surfactants include sulfate ester salts, α-olefin sulfonates, sulfosuccinates, N-acyl amino acid salts, and acylated methyl taurine salts. Examples of the amphoteric surfactants include betaine acetate amphoteric surfactants and imidazoline amphoteric surfactants.

[0052] Examples of anti-inflammatory agents include aminocaproic acid, allantoin and its derivatives, ascorbic acid and its salts, azulene, azulene sulfonate, dihydrocholesterol, epidihydrocholesterol, tocopherol acetate, tocopherol nicotinate, lysozyme chloride, and the like.

[0053] Examples of anti-tartar agents include phosphates, polyphosphates, methoxyethylene maleic anhydride copolymers, zinc chloride, zinc organic acids, edetates, and the like.

[0054] Examples of pH adjusters include sodium phosphate salts such as monosodium phosphate and disodium phosphate, sodium citrate, citric acid, glucono-δ-lactone, sodium gluconate, sodium acetate, sodium carbonate, sodium bicarbonate, sodium hydroxide, and hydrates thereof.

[0055] Examples of the enzyme include protease, dextranase, amylase, mutanase, and lysozyme.

[0056] Examples of the ion source include fluoride ion sources such as sodium fluoride, monofluorophosphate, and stannous fluoride, and calcium ion sources such as calcium phosphate, calcium gluconate, and hydroxyapatite.

[0057] Examples of preservatives include paraoxybenzoic acid esters, sodium benzoate, potassium sorbate, and phenoxyethanol.

[0058] Examples of fragrances include natural essential oils such as peppermint oil, spearmint oil, mint oil, eucalyptus oil, clove oil, thyme oil, rosemary oil, lemon oil, ginger oil, lime oil, cassia oil, cardamom oil, lavender oil, and cinnamon oil; fragrance components such as l-menthol, l-carvone, carvacrol, eugenol, anethole, 1,8-cineole, thymol, vanillin, pinene, 3-l-menthoxypropane-1,2-diol, and methyl salicylate; mixtures of these, natural fragrances, blended fragrances, and synthetic fragrances.

[0059] Examples of herbal medicines include horsetail extract, hawthorn extract, witch hazel extract, peony extract, white birch extract, sage extract, seaweed extract, Phellodendron bark extract, Angelica extract, carrot extract, and fennel extract.

[0060] Examples of pigments include Blue No. 1, Blue No. 2, Blue No. 201, Yellow No. 4, Yellow No. 5, Yellow No. 202(1), Yellow No. 203, Red No. 3, Red No. 102, Red No. 104, Red No. 105, Green No. 3, and Green No. 201.

[0061] Moisturizing ingredients include glycerin, heparinoids (glycosaminoglycans), hyaluronic acid and its salts, petrolatum, and the like.

[0062] Examples of cell-activating ingredients include N-acetylglucosamine, γ-aminobutyric acid, acetylglucosamine, aminobutyric acid, allantoin, chlorella extract, white birch extract, white birch bark extract, taiso extract, centella asiatica extract, corn extract, jujube fruit extract, Job's tears seed extract, retinol palmitate, Belamcanda chinensis extract, Belamcanda chinensis extract, bilberry leaf extract, yuzu seed extract, coix seed extract, retinol, etc.

[0063] Examples of vitamins include fat-soluble vitamins such as vitamin A, vitamin D, vitamin E, and vitamin K, and water-soluble vitamins such as vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, pantothenic acid, folic acid, biotin, and vitamin C.

[0064] Amino acids include valine, isoleucine, leucine, methionine, lysine (lysine), phenylalanine, tryptophan, threonine (threonine), histidine, arginine, glycine, alanine, serine, tyrosine, cysteine, asparagine, glutamine, proline, aspartic acid, glutamic acid, and the like.

[0065] When the composition of the present invention contains optional components, the content of at least one of the components (A) and (B) in the composition is not particularly limited as long as it is an effective amount, but is preferably 0.001 to 1 w / v%.

[0066] The composition of the present invention can be prepared by mixing at least one of the components (A) and (B), the component (C) and / or any optional components, if necessary, with heating, and stirring the components uniformly.

[0067] The administration or ingestion form of the agent or composition of the present invention is not particularly limited, but for example, oral administration such as oral administration or sublingual administration, oral ingestion, application to epithelial mucosa such as the oral cavity or skin, application by patch or spray, etc. are preferred.

[0068] The composition of the present invention can be prepared in the form of, for example, liquid, gel, paste, cream, solid such as tablet, powder such as granule or fine granule, capsule, soft capsule, etc. Among them, liquid preparations are preferred because the active ingredient can be uniformly applied to the epithelial mucosa and skin of the oral cavity, etc., and examples of liquid preparations include mouthwash (also called gargle), liquid dentifrice, mouthwash, mouth spray, lotion, body spray, etc., and mouthwash (also called gargle), liquid dentifrice, mouthwash, mouth spray, etc., which can be uniformly applied to the oral cavity, are particularly preferred.

[0069] In this specification, the term "liquid" refers to a material that has fluidity and has a viscosity of 20 mPa s or less, preferably 10 mPa s or less, at 20° C. Here, the viscosity can be measured using a commercially available rotational viscometer (for example, a B-type viscometer manufactured by Toki Sangyo Co., Ltd.).

[0070] As described above, the agent consisting of at least one of the components (A) and (B) and the composition containing at least one of the components (A) and (B) as an active ingredient can suppress the expression of one or more genes selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene. Furthermore, the agent consisting of at least one of the components (A) and (B) and the composition containing at least one of the components (A) and (B) as an active ingredient can promote the expression of the Pla2g2f gene in addition to suppressing the expression of the above genes.

[0071] In the present specification, the Pla2g2f gene includes the human-derived PLA2G2F gene (NCBI Gene accession number 64600) and a homologue in the human-derived PLA2G2F gene. When aligned with the base sequence of the human-derived PLA2G2F gene, the homologue in the human-derived PLA2G2F gene has a base sequence identity of preferably 70% or more, more preferably 80% or more, and even more preferably 90% or more. The homologue in the human-derived PLA2G2F gene may be a human-derived gene or a gene derived from an animal other than human. Examples of animals other than human include hamsters, dogs, cats, rabbits, etc.

[0072] The expression inhibitor of the Arid5a gene and the composition for inhibiting the expression of the gene can prevent and / or treat diseases caused by overexpression of the Arid5a gene. The expression of the Arid5a gene is inhibited by administering or ingesting an agent consisting of at least one of the components (A) and (B) or a composition containing at least one of the components (A) and (B) as an active ingredient. Therefore, the administration or ingestion of the agent or the composition can prevent and / or treat diseases caused by overexpression of the Arid5a gene, and is considered to be particularly useful for prevention.

[0073] Arid5a promotes both the mRNA stability and translation of IκBξ, a member of the IκB family (Nilesh Amatya et al., “IL-17 integratesmultiple self-reinforcing, feed-forward mechanisms through the RNA binding protein Arid5a”, SCIENCE SIGNALING, 2018, 11, eaat4617) and inhibits the degradation of IL-6 mRNA (K Masuda et al., “Arid5a controls IL-6 mRNAstability, which contributes to elevation of IL-6 level in vivo”, PNAS., 2013, 110 p.9409-9414.), making it highly likely that it is involved in biological reactions involving innate immunity and is therefore effective against these related diseases. Therefore, diseases caused by overexpression of the Arid5a gene include inflammatory diseases such as atopic dermatitis, ulcerative diseases, allergic diseases, bone destructive diseases, autoimmune diseases such as rheumatism, neurodegenerative diseases, and cancer.

[0074] That is, suppression of excessive Arid5a gene expression is effective for preventing and / or treating inflammatory diseases such as atopic dermatitis, ulcerative diseases, allergic diseases, bone destructive diseases, autoimmune diseases such as rheumatism, neurodegenerative diseases, and cancer, and is particularly effective for prevention. Since degradation of IκBξ and mRNA of inflammatory cytokines such as IL-6 present downstream of IκBξ is promoted, prevention and / or treatment of inflammatory diseases and autoimmune diseases is preferred.

[0075] An inhibitor of the expression of the Rac2 gene and a composition for inhibiting the expression of the gene can prevent and treat diseases caused by overexpression of the Rac2 gene. Rac2 gene expression is suppressed by administering or ingesting an agent consisting of at least one of the components (A) and (B) or a composition containing at least one of the components (A) and (B) as an active ingredient. Therefore, administration or ingestion of the agent or composition can prevent and / or treat diseases caused by overexpression of the Rac2 gene, and is considered to be particularly useful for prevention.

[0076] In activated macrophages, Rac2 interacts with NOS2 and activates NOS2 (Teresa Kuncewicz et al., “Specific association of nitric oxide synthase-2 with Rac isoforms in activated murinemacrophages”, Am J Physiol Renal Physiol. 2001, 281, p.F326-336.), so suppression of the Rac2 gene is considered to be effective for diseases in which excessive generation of reactive oxygen species is the main or secondary cause. Therefore, diseases caused by overexpression of the Rac2 gene include inflammatory diseases such as atopic dermatitis, ulcerative diseases, allergic diseases, bone destructive diseases, autoimmune diseases such as rheumatism, neurodegenerative diseases, and cancer.

[0077] In other words, suppression of excessive Rac2 gene expression is effective in preventing and / or treating inflammatory diseases such as atopic dermatitis, ulcerative diseases, allergic diseases, bone destructive diseases, autoimmune diseases such as rheumatism, neurodegenerative diseases, and cancer, and is particularly effective in prevention, and has the effect of suppressing the overproduction of reactive oxygen species, so is preferable for the prevention and / or treatment of inflammatory diseases and ulcerative diseases.

[0078] The Clic4 gene expression inhibitor and the composition for inhibiting said gene expression can prevent and / or treat diseases caused by overexpression of the Clic4 gene. Clic4 gene expression is inhibited by administering or ingesting an agent consisting of at least one of the components (A) and (B) or a composition containing at least one of the components (A) and (B) as an active ingredient. Therefore, administration or ingestion of the agent or composition can prevent and / or treat diseases caused by overexpression of the Clic4 gene, and is considered to be particularly useful for prevention.

[0079] Clic4 is not only present in the cytoplasm but also a component of the inner mitochondrial membrane, and maintaining the expression level of Clic4 is essential for keratinocyte survival (Fernandez-Salas, E., et al., “mtCLIC / CLIC4, an organellular chloride channel protein, is increased by DNA damage andparticipates in the apoptotic response to p53”, Mol. Cell. Biol., 2002, 22, p.3610-3620.). Increased expression of Clic4 in differentiating keratinocytes may be involved in the terminal phase of differentiation, where keratinocytes undergo cell cycle arrest, karyorupture, and die. In fact, it has been reported that even a slight increase in Clic4 expression significantly increases the differentiation response of keratinocytes (Kwang S Suh, et al., "CLIC4 mediates and is required for Ca2+-induced keratinocyte differentiation", J Cell Sci., 2007, 120, p.2631-2640.). Therefore, diseases caused by overexpression of the Clic4 gene include inflammatory diseases such as atopic dermatitis, ulcerative diseases, allergic diseases, bone destructive diseases, autoimmune diseases such as rheumatism, and neurodegenerative diseases.

[0080] In other words, suppression of excessive Clic4 gene expression is effective in preventing and / or treating inflammatory diseases such as atopic dermatitis, ulcerative colitis, allergic diseases, bone destructive diseases, autoimmune diseases such as rheumatism, and neurodegenerative diseases, and is particularly effective in prevention.

[0081] The keratin gene family expression inhibitor and the composition for inhibiting gene expression can prevent and / or treat diseases caused by overexpression of the keratin gene family. By administering or ingesting an agent consisting of at least one of the components (A) and (B) or a composition containing at least one of the components (A) and (B) as an active ingredient, the expression of the keratin gene family is inhibited. Therefore, by administering or ingesting the agent or composition, diseases caused by overexpression of the keratin gene family can be prevented and / or treated, and are considered to be particularly useful for prevention.

[0082] It has been reported that overexpression of Krt16 disrupts the normal keratin profile and inhibits the terminal differentiation process of epithelium. In addition, there are research reports that increased expression of inflammatory cytokines such as Interleukin-17 (IL-17) and Interleukin-22 (IL-22) promotes nuclear translocation of Nrf2, increases expression of Krt6, Krt16, and Krt17, and promotes proliferation of keratinocytes, resulting in psoriasis (Luting Yang, et al., “Nrf2 Promotes Keratinocyte Proliferation in Psoriasis through Up-Regulation of Keratin 6, Keratin 16, and Keratin 17”, J Invest Dermatol., 2017,137, p.2168-2176.). Therefore, it is believed that preventing overexpression of the keratin gene family contributes to the prevention and / or treatment of inflammatory diseases such as atopic dermatitis, allergic diseases, psoriasis, etc.

[0083] That is, suppression of excessive expression of the keratin gene family is effective in preventing and / or treating inflammatory diseases such as atopic dermatitis, allergic diseases, psoriasis, and the like, and is particularly effective in preventing such diseases.

[0084] The Eps15 gene expression inhibitor and the composition for inhibiting said gene expression can prevent and / or treat diseases caused by overexpression of the Eps15 gene. Eps15 gene expression is inhibited by administering or ingesting an agent consisting of at least one of the components (A) and (B) or a composition containing at least one of the components (A) and (B) as an active ingredient. Therefore, administration or ingestion of the agent or composition can prevent and / or treat diseases caused by overexpression of the Eps15 gene, and is considered to be particularly useful for prevention.

[0085] Eps15 contributes to the receptor-mediated endocytosis of epidermal growth factor, and since endocytosis is a major biological reaction involving innate immunity, suppression of the Eps15 gene is considered to be effective in preventing and / or treating these related diseases. Diseases caused by overexpression of the Eps15 gene include inflammatory diseases such as atopic dermatitis, ulcerative colitis, allergic diseases, osteoclastic diseases, autoimmune diseases such as rheumatism, and neurodegenerative diseases.

[0086] In other words, suppression of excessive Eps15 gene expression is effective in preventing and / or treating inflammatory diseases such as atopic dermatitis, ulcerative inflammation, allergic diseases, bone destructive diseases, autoimmune diseases such as rheumatism, and neurodegenerative diseases, and is particularly effective in preventing and contributing to endocytosis, so is preferable for the prevention and / or treatment of inflammatory diseases and autoimmune diseases.

[0087] The agent of one embodiment of the present invention and the composition of one embodiment of the present invention can prevent and / or treat diseases caused by the expression of one or more genes selected from the group consisting of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene, as well as prevent diseases caused by deficiency or reduced expression of Pla2g2f gene. Pla2g2f gene expression can be promoted by administering or ingesting the agent consisting of at least one of the components (A) and (B) or the composition containing at least one of the components (A) and (B) as an active ingredient. Therefore, it is considered that diseases caused by deficiency or reduced expression of Pla2g2f gene can be prevented by administering or ingesting the agent or the composition.

[0088] Since the barrier function of the skin of Pla2g2f knockout mice is impaired (Kei Yamamoto et al., “The role of group IIF-secreted phospholipase A2 in epidermal homeostasis and hyperplasia”, J. Exp. Med., 2015, 212, p.1901-1919.), diseases caused by a deficiency or reduced expression of the Pla2g2f gene include inflammatory diseases such as atopic dermatitis, ulcerative diseases, allergic diseases, and autoimmune diseases such as rheumatism.

[0089] In other words, promoting the expression of the Pla2g2f gene is effective in preventing inflammatory diseases such as atopic dermatitis, ulcerative diseases, allergic diseases, and autoimmune diseases such as rheumatism. In particular, since the Pla2g2f gene is important for maintaining the skin's barrier function, preventing inflammatory diseases and ulcerative diseases is preferable.

[0090] Examples of inflammatory diseases include rheumatoid arthritis, systemic lupus erythematosus, scleroderma, bronchial asthma, asthmatic bronchitis, diffuse interstitial pneumonia, chronic obstructive pulmonary disease, ulcerative colitis, Crohn's disease, acute hepatitis, chronic hepatitis, fulminant hepatitis, autoimmune hepatitis, primary biliary cirrhosis, primary sclerosing cholangitis, alcoholic hepatitis, nonalcoholic steatohepatitis, liver cirrhosis, peripheral neuritis, ankylosing spondylitis, eczema (acute, subacute, chronic), contact dermatitis, solar (ultraviolet) dermatitis, radiation dermatitis, atopic dermatitis, seborrheic dermatitis, psoriasis, lichen planus, erythema, rosacea, urticaria, alopecia areata, and pemphigus. Examples of the oral mucositis (stomatitis) including acne vulgaris, pemphigoid, erythroderma, common acne, bedsore, wound, burn, conjunctivitis, keratitis, scleritis, acute and chronic otitis media, hay fever, sinusitis, laryngitis, esophagitis, aphthous stomatitis, recurrent aphthous stomatitis, catarrhal stomatitis, herpes stomatitis, candidal stomatitis, drug-induced stomatitis, and radiation stomatitis, oral lichen planus, glossitis, acute and chronic sialadenitis, angular stomatitis, cheilitis, Behcet's disease, multiple sclerosis, diabetes mellitus, atherosclerosis, pancreatitis, and chronic heart failure, among which stomatitis, oral lichen planus, ulcerative colitis, and pemphigus are preferred.

[0091] Examples of ulcerative diseases include stomatitis, esophagitis, esophageal ulcer, gastritis, ulcerative colitis, and gastric ulcer, with stomatitis and gastric ulcer being preferred.

[0092] Examples of allergic diseases include allergic conjunctivitis, allergic rhinitis, allergic granulomatous vasculitis, contact dermatitis, and the like, with allergic conjunctivitis being preferred.

[0093] Examples of bone destructive diseases include periodontal disease, osteoporosis, rheumatoid arthritis, osteonecrosis of the jaw, and necrosis of the femoral head, with periodontal disease being preferred.

[0094] Examples of autoimmune diseases include Behcet's disease, systemic lupus erythematosus, chronic discoid lupus erythematosus, multiple sclerosis (systemic sclerosis, progressive systemic sclerosis), scleroderma, polymyositis, dermatomyositis, periarteritis nodosa (polyarteritis nodosa, microscopic polyangiitis), aortitis syndrome (Takayasu's arteritis), malignant rheumatoid arthritis, rheumatoid arthritis, juvenile idiopathic arthritis, spondyloarthritis, mixed connective tissue disease, Sjogren's syndrome, adult Still's disease, vasculitis, hypersensitivity vasculitis, rheumatoid arthritis, and rheumatoid arthritis. vasculitis, large vessel vasculitis, ANCA-associated vasculitis (e.g., granulomatosis with polyangiitis and eosinophilic granulomatosis with polyangiitis), Cogan's syndrome, RS3PE syndrome, temporal arteritis, polymyalgia rheumatica, fibromyalgia, antiphospholipid syndrome, eosinophilic fasciitis, IgG4-related disease (e.g., primary sclerosing cholangitis, autoimmune pancreatitis, etc.), Guillain-Barré syndrome, myasthenia gravis, chronic atrophic gastritis, autoimmune hepatitis, primary biliary cirrhosis, Goodpasture's syndrome, rapidly progressive glomerulonephritis, corpus nephritis, megaloblastic anemia, autoimmune hemolytic anemia, pernicious anemia, autoimmune neutropenia, idiopathic thrombocytopenic purpura, Graves' disease (Graves' disease (hyperthyroidism)), Hashimoto's disease, autoimmune adrenal insufficiency, primary hypothyroidism, Addison's disease (chronic adrenal insufficiency), idiopathic Addison's disease, type I diabetes mellitus, slowly progressive type I diabetes mellitus (adult latent autoimmune diabetes mellitus), localized scleroderma, psoriasis, bullous pemphigoid, pemphigus, pemphigoid, herpes gestationis, linear IgA bullous dermatosis disease, epidermolysis bullosa acquisita, neuromyelitis optica, chronic inflammatory demyelinating polyneuropathy, multifocal motor neuropathy, sarcoidosis, giant cell arteritis, amyotrophic lateral sclerosis, Harada's disease, autoimmune optic neuropathy, idiopathic azoospermia, inflammatory bowel disease (e.g., ulcerative colitis, Crohn's disease), celiac disease, ankylosing spondylitis, severe asthma, chronic urticaria, transplantation immunity, familial Mediterranean fever, eosinophilic sinusitis, dilated cardiomyopathy, systemic mastocytosis, or inclusion body myositis, and Behcet's disease is preferred.

[0095] Examples of neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, frontotemporal dementia, brain injury, spinal cord injury, hypoxia, convulsion, and traumatic brain injury, with Alzheimer's disease being preferred.

[0096] Examples of cancer include oral squamous cell carcinoma, breast cancer, ovarian cancer, colon cancer, lung cancer, prostate cancer, head and neck cancer, lymphoma, uveal malignant melanoma, thymoma, mesothelioma, esophageal cancer, gastric cancer, duodenal cancer, hepatocellular carcinoma, bile duct cancer, gallbladder cancer, pancreatic cancer, renal cell cancer, renal pelvis and ureter cancer, bladder cancer, penile cancer, testicular cancer, uterine cancer, vaginal cancer, vulvar cancer, skin cancer (e.g., malignant melanoma), malignant bone tumor, soft tissue sarcoma, chondrosarcoma, leukemia, myelodysplastic syndrome, brain tumor, and multiple myeloma, with oral squamous cell carcinoma being preferred.

[0097] Psoriasis includes psoriasis vulgaris, psoriatic arthritis, guttate psoriasis, erythrodermic psoriasis, and pustular psoriasis, with psoriasis vulgaris being preferred.

[0098] Another embodiment of the present invention includes a method for suppressing the expression of one or more genes selected from the group consisting of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene, which comprises administering or ingesting at least one of (A) glycyrrhizinic acid and (B) tranexamic acid to a subject. Another embodiment includes a method for promoting Pla2g2f gene expression in addition to suppressing the expression of one or more genes selected from the group consisting of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene by the administration or ingestion. Here, the subject may be other than a human individual.

[0099] By administering or ingesting at least one of the components (A) and (B) to a subject, the expression of at least one gene selected from the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene is suppressed, and the disease caused by the Arid5a gene expression, the disease caused by the Rac2 gene expression, the disease caused by the Clic4 gene expression, the disease caused by the keratin gene family expression, and the disease caused by the Eps15 gene expression can be prevented and / or treated. The administration or ingestion can also prevent diseases caused by the deficiency or reduced expression of the Pla2g2f gene. When used to prevent diseases caused by the Arid5a gene expression, the disease caused by the Rac2 gene expression, the Clic4 gene expression, the disease caused by the keratin gene family expression, and the disease caused by the Eps15 gene expression, the administration or ingestion is performed before the onset of the disease, but may be performed after the onset, and when used to prevent the recurrence of the disease, the administration or ingestion may be performed after the disease is cured. When used to treat the above diseases, the above administration or ingestion is carried out after the onset of the above diseases. The number of administrations or ingestions is not particularly limited, but it is preferable to administer or ingestion three or more times a day for one month or more. EXAMPLES

[0100] The present invention will be specifically described below by way of examples, but the present invention is not limited to the following examples as long as it does not depart from the gist of the present invention.

[0101] <Test Example 1: Evaluation of gene expression suppression and promotion effects (1)> (Example A-1) The following test was carried out using physiological saline containing 0.01 (w / v)% dipotassium glycyrrhizinate (0.0065 (w / v)% in terms of glycyrrhizinic acid). (Example A-2) A similar test was performed using saline containing 0.05 (w / v)% tranexamic acid. (Control Example A-1) A similar test was performed using saline. (control) None of the samples described in Examples A-1 and A-2 and Control Example A-1 were applied, and one hamster per group was raised for 6 days, after which the following test method was performed in step I.2-1).

[0102] (Test Method) I. Animal Experiments 1) 50μL of the above sample was applied to the right buccal mucosa of each hamster (Syrian, male, 5 weeks old) twice a day. The first and second applications were spaced at least 3 hours apart. This procedure was repeated for 4 consecutive days. On the fifth day, the same application was repeated once. 2-1) In the group in which the expression of the Pla2g2f gene was quantified, the hamsters were euthanized with carbon dioxide gas after 1), while in the control group, 1) was not performed. 2-2) In the group in which the expression of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene was quantified, after the application on the 5th day in 1), a Φ6mm filter paper soaked in 30(w / v)% acetic acid solution was applied to the right buccal mucosa of the above hamsters under anesthesia for 10 seconds to induce stomatitis. The hamsters were euthanized with carbon dioxide gas 3, 12, and 24 hours after the induction of stomatitis. 3) The surrounding tissue, including the right buccal mucosa, was collected.

[0103] II. RNA extraction, purification, and quality check 1) RNA was extracted and purified from the tissue and treated with DNase using TRI Reagent (Molecular Research Center), Purelink RNA Mini Kit (Thermo Fisher Scientific), and Purelink DNase (Thermo Fisher Scientific) according to the protocol. 2) Using an Agilent 4150 TapeStation system (Agilent Technologies), it was confirmed that the RINe (RNA Integrity Number equivalent) values ​​of all RNA samples were 7 or higher. 3) RNA sequencing was performed under the conditions in Table 1.

[0104] [Table 1]

[0105] III. RNA sequencing data processing i) Expression quantification using raw data 1) We obtained the cDNA sequence of the golden hamster (MesAur1.0) from the Ensembl genome browser (release 105) and created a salmon index. 2) After trimming the RNA sequence reads with fastp (v0.23.2), the expression of the Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, Eps15 gene, and Pla2g2f gene was quantified using salmon (v1.6.0). 3) I created a table of TPM (transcripts per million) values ​​using tximport (v1.22.0).

[0106] ii) Calculation of expression ratios and addition of annotation information The expression level ratios (TPM(sample)+1 / TPM(control)+1) of Example A-1, Example A-2, and Control Example A-1 relative to the control were calculated.

[0107] The results regarding the expression ratio of the Arid5a gene in the group of mice euthanized 3 hours after the induction of stomatitis are shown in Figure 1. Compared to the control, the expression level of the Arid5a gene was significantly higher in Control Example A-1. On the other hand, it was shown that the expression level of the Arid5a gene was suppressed more in Examples A-1 and A-2 than in Control Example A-1. In other words, it was shown that the expression of the Arid5a gene was suppressed by administration of the agent or composition according to one embodiment of the present invention.

[0108] The results of the expression ratio of the Rac2 gene in the group of mice euthanized 3 hours after the induction of stomatitis are shown in Figure 2. Compared to the control, the expression level of the Rac2 gene was significantly higher in Control Example A-1. On the other hand, it was shown that the expression level of the Rac2 gene was suppressed more in Examples A-1 and A-2 than in Control Example A-1. In other words, it was shown that the expression of the Rac2 gene was suppressed by administration of the agent or composition according to one embodiment of the present invention.

[0109] The results regarding the expression ratio of the Clic4 gene in the group of mice euthanized 3 hours after induction of stomatitis are shown in Figure 3. Compared to the control, the expression level of the Clic4 gene was significantly higher in control example A-1. On the other hand, it was shown that the expression level of the Clic4 gene was suppressed more in Examples A-1 and A-2 than in control example A-1. In other words, it was shown that the expression of the Clic4 gene was suppressed by administration of the agent or composition of one embodiment of the present invention.

[0110] The results of the expression ratio of the Krt1 gene in the group of mice euthanized 3 hours after the induction of stomatitis are shown in FIG. 4A. The results of the expression ratios of the Krt14 gene, the Krt16 gene, and the Krt17 gene in the group of mice euthanized 12 hours after the induction of stomatitis are shown in FIG. 4B, FIG. 4C, and FIG. 4D, respectively. In all of FIG. 4A, FIG. 4B, FIG. 4C, and FIG. 4D, the expression level of the keratin gene family was significantly higher in Control Example A-1 than in the control. On the other hand, it was shown that the expression level of the keratin gene family was suppressed more in Examples A-1 and A-2 than in Control Example A-1. That is, it was shown that the expression of the keratin gene family was suppressed by administration of the agent or composition according to one embodiment of the present invention.

[0111] FIG. 5 shows the results of the expression ratio of the Eps15 gene in the group of mice euthanized 24 hours after induction of stomatitis. Compared to the control, the expression level of the Eps15 gene was significantly higher in Control Example A-1. On the other hand, it was shown that the expression level of the Eps15 gene was suppressed more in Examples A-1 and A-2 than in Control Example A-1. In other words, it was shown that the expression of the Eps15 gene was suppressed by administration of the agent or composition according to one embodiment of the present invention.

[0112] The results regarding the expression ratio of the Pla2g2f gene in the group of individuals in which stomatitis was not induced are shown in Figure 6. It was shown that the expression ratio of the Pla2g2f gene was significantly higher in Examples A-1 and A-2 than in Control Example A-1 and the control. In other words, it was shown that the administration of the agent or composition according to one aspect of the present invention promotes the expression of the Pla2g2f gene.

[0113] <Test Example 2: Evaluation of gene expression suppression and promotion effects (2)> The liquid samples of Example A-1, Example A-2 and Control Example A-1 in Test Example 1 were subjected to the following tests. As a control, right buccal mucosa tissue from a hamster to which none of the samples described in Examples A-1 and A-2 and Control Example A-1 had been applied was collected and subjected to step 2) of the following test method.

[0114] (Test Method) 1) The right buccal mucosa of a hamster (Syrian, male, 5 weeks old) that had been kept for one week was 50 μL of the above sample was applied to each mouse twice a day for four consecutive days, with an interval of at least three hours between the first and second applications. 2) On the fifth day, after the sample was applied once, three mice from each of the groups applied with Example A-1, Example A-2, and Control A-1 were euthanized and the right buccal mucosa tissue was collected. At this time, the control group was also euthanized in the same manner and the right buccal mucosa tissue was collected. 3) In the remaining groups of 6 hamsters, which were treated with Example A-1, Example A-2, and Control A-1, a Φ6 mm filter paper impregnated with 30 (w / v)% acetic acid aqueous solution was applied to the right buccal mucosa of the hamsters under anesthesia for 10 seconds to induce stomatitis. Three or 24 hours after induction of stomatitis, the right buccal mucosa tissue including the stomatitis induction site was collected from 3 hamsters in each group. 4) Total RNA was extracted from the collected tissues using TRIzol reagent (Invitrogen) according to the protocol. 5) Reverse transcription was performed using RNA as a template according to the protocol for ReverTra Ace (registered trademark) (manufactured by Toyobo Co., Ltd.). 6) Real-time PCR was performed using the ΔΔCt method under the following conditions with cDNA as a template, following the protocol of FastStart Essential DNA Green Master (Roche).

[0115] (Primer information) SEQ ID NO: 1: F-Pla2g2f: 5'-CAAGCAGGGTGGCAGTAGAA-3' SEQ ID NO: 2: R-Pla2g2f: 5'-CACGCAGTGACCCCTTAGTC-3' SEQ ID NO: 3: F-Arid5a: 5'-GTTGTTGGGACCGCCTGATA-3' SEQ ID NO: 4: R-Arid5a: 5'-CAGGCTTTGGGCTTGGGATA-3' SEQ ID NO: 5: F-Rac2: 5'-AAATGAGGAGCTCAGCCCAC-3' SEQ ID NO: 6: R-Rac2: 5'-TGTCTGCAGTGGTTCTGGTC-3' SEQ ID NO: 7: F-Clic4: 5'-TGGGATTGCGTCTTTCGTGA-3' SEQ ID NO: 8: R-Clic4: 5'-ATCTCACCCAGCTTCAGTGC-3' SEQ ID NO: 9: F-Krt1: 5'-CTCCTTCATCGACAAGGTGCG-3' SEQ ID NO: 10: R-Krt1: 5'-TGGTTGTTGTATCCACCTGCT-3' SEQ ID NO: 11: F-Krt5: 5'-CTTCAGCTCGGTGTCTCGAA-3' SEQ ID NO: 12: R-Krt5: 5'-TACAGGCTCCGGCTACCATA-3' SEQ ID NO: 13: F-Krt14: 5'-CAGTCCCAGCTCAGCATGAA-3' SEQ ID NO: 14: R-Krt14: 5'-TGGGACAGCTGCATGCAATA-3' SEQ ID NO: 15: F-Krt16: 5'-ACAAGATCCTGCTGGACGTG-3' SEQ ID NO: 16: R-Krt16: 5'-TTGCGAGGAGATTTGGGCAT-3' SEQ ID NO: 17: F-Eps15: 5'-CAGTGGGCCAATGTCGTACT-3' SEQ ID NO: 18: R-Eps15: 5'-TCTTTGTTTCAAGCGCAGGC-3' SEQ ID NO: 19: F-GAPDH: 5'-ACAGTCAAGGCTGAGAACGG-3' SEQ ID NO: 20: R-GAPDH: 5'-CAGGCGACATGTGAGATCCA-3'

[0116] The primers having the nucleotide sequences shown in SEQ ID NOs: 1 and 2 are based on the hamster-derived Pla2g2f gene mRNA sequence (NCBI Nucleotide Accession No. XM_021233581.2), the primers having the nucleotide sequences shown in SEQ ID NOs: 3 and 4 are based on the hamster-derived Arid5a gene mRNA sequence (NCBI Nucleotide Accession No. XM_040729415.1), the primers having the nucleotide sequences shown in SEQ ID NOs: 5 and 6 are based on the hamster-derived Rac2 gene mRNA sequence (NCBI Nucleotide Accession No. XM_005066960.4), the primers having the nucleotide sequences shown in SEQ ID NOs: 7 and 8 are based on the hamster-derived Clic4 gene mRNA sequence (NCBI Nucleotide Accession No. XM_005079597.4), and the primers having the nucleotide sequences shown in SEQ ID NOs: 9 and 10 are based on the hamster-derived Krt1 gene mRNA sequence (NCBI Nucleotide Accession No. XM_005079597.4). XM_005067285.4), the primers having the nucleotide sequences shown in SEQ ID NOs: 11 and 12 are based on the hamster-derived Krt5 gene mRNA sequence (NCBI Nucleotide Accession No. XM_005067276.4), the primers having the nucleotide sequences shown in SEQ ID NOs: 13 and 14 are based on the hamster-derived Krt14 gene mRNA sequence (NCBI Nucleotide Accession No. XM_005087369.4), the primers having the nucleotide sequences shown in SEQ ID NOs: 15 and 16 are based on the hamster-derived Krt16 gene mRNA sequence (NCBI Nucleotide Accession No. XM_040749688.1), and the primers having the nucleotide sequences shown in SEQ ID NOs: 17 and 18 are based on the hamster-derived Eps15 gene mRNA sequence (NCBI Nucleotide Accession No. XM_040756977.1), and primers having the nucleotide sequences shown in SEQ ID NOs: 19 and 20 were designed based on the hamster-derived GAPDH gene mRNA sequence (NCBI Nucleotide Accession No. DQ403055.1). Furthermore, the primers having the nucleotide sequences shown in SEQ ID NOs: 15 and 16 can detect both Krt16 and Krt17.

[0117] (PCR conditions) 1 cycle at 95℃ for 10 minutes 45 cycles of 95°C for 15 seconds, 59°C for 20 seconds, and 72°C for 40 seconds 1 cycle of 72℃ for 5 minutes

[0118] Results regarding the expression level (expression level ratio) of each gene relative to the expression level of GAPDH are shown in Figures 7 to 12. Figure 7 shows the expression level of the Pla2g2f gene relative to the expression level of GAPDH (Pla2g2f / GAPDH), Figure 8 shows the expression level of the Arid5a gene relative to the expression level of GAPDH (Arid5a / GAPDH), Figure 9 shows the expression level of the Rac2 gene relative to the expression level of GAPDH (Rac2 / GAPDH), Figure 10 shows the expression level of the Clic4 gene relative to the expression level of GAPDH (Clic4 / GAPDH), and Figure 11A shows the expression level of the Krt1 gene relative to the expression level of GAPDH. 11B shows the expression level of the Krt5 gene relative to the expression level of GAPDH (Krt5 / GAPDH), FIG. 11C shows the expression level of the Krt14 gene relative to the expression level of GAPDH (Krt14 / GAPDH), FIG. 11D shows the expression level of the Krt16·17 gene relative to the expression level of GAPDH (Krt16·17 / GAPDH), and FIG. 12 shows the expression level of the Eps15 gene relative to the expression level of GAPDH (Eps15 / GAPDH). The results regarding the expression level of the Krt16 gene shown in FIG. 11D can also be interpreted as the results regarding the expression level of the Krt17 gene.

[0119] In Figures 7 to 12, Control Example A-1 (0h), Example A-1 (0h) and Example A-2 (0h) all show the results of hamsters euthanized without inducing stomatitis in Test Method 2). Control Example A-1 (3h), Example A-1 (3h) and Example A-2 (3h) all show the results of hamsters euthanized 3 hours after inducing stomatitis in Test Method 3). Control Example A-1 (24h), Example A-1 (24h) and Example A-2 (24h) all show the results of hamsters euthanized 24 hours after inducing stomatitis in Test Method 3). All groups have n=3.

[0120] As shown in FIG. 7, it was confirmed that application of component (A) or component (B) increased the mRNA expression of Pla2g2f gene in the buccal mucosa tissue of hamsters before the induction of stomatitis. As shown in Figures 8 to 12, it was confirmed that application of component (A) or component (B) suppressed the mRNA expression of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene in hamsters with stomatitis induced, compared to control example A-1.

[0121] From this, the expression of each gene in Test Examples 1 and 2 showed similar trends.

[0122] <Test Example 3: Evaluation of gene expression suppression and promotion effects (3)> The following test was carried out in the same manner as in Test Example 2 above, except that the following Examples A-3 to A-9 and the above-mentioned Control Example A-1 were used instead of Examples A-1, A-2 and Control Example A-1, and the group to which Control Example A-1 was applied in Test Example 2 2) was not euthanized. As a control, right buccal mucosa tissue from a hamster to which none of the samples described in Examples A-3 to A-9 and Control Example A-1 below had been applied was collected and subjected to the same steps as in Test Example 2 above.

[0123] (Example A-3) The following test was carried out using physiological saline containing 0.005 (w / v)% dipotassium glycyrrhizinate (0.00325 (w / v)% in terms of glycyrrhizinic acid). (Example A-4) The following test was carried out using physiological saline containing 0.478 (w / v)% dipotassium glycyrrhizinate (0.31 (w / v)% in terms of glycyrrhizinic acid). (Example A-5) A similar test was performed using saline containing 0.003 (w / v)% tranexamic acid. (Example A-6) A similar test was performed using saline containing 1 (w / v)% tranexamic acid. (Example A-7) A similar test was performed using saline containing 0.005 (w / v)% dipotassium glycyrrhizinate (0.00325 (w / v)% in terms of glycyrrhizinic acid) and 1 (w / v)% tranexamic acid. (Example A-8) A similar test was performed using saline containing 0.478 (w / v)% dipotassium glycyrrhizinate (0.31 (w / v)% in terms of glycyrrhizinic acid) and 0.003 (w / v)% tranexamic acid. (Example A-9) A similar test was performed using saline containing 0.01 (w / v)% dipotassium glycyrrhizinate (0.0065 (w / v)% in terms of glycyrrhizinic acid) and 0.05 (w / v)% tranexamic acid.

[0124] Results regarding the expression level (expression level ratio) of each gene relative to the expression level of GAPDH are shown in Figures 13 to 18. Figure 13 shows the expression level of the Pla2g2f gene relative to the expression level of GAPDH (Pla2g2f / GAPDH), Figure 14 shows the expression level of the Arid5a gene relative to the expression level of GAPDH (Arid5a / GAPDH), Figure 15 shows the expression level of the Rac2 gene relative to the expression level of GAPDH (Rac2 / GAPDH), Figure 16 shows the expression level of the Clic4 gene relative to the expression level of GAPDH (Clic4 / GAPDH), and Figure 17A shows the expression level of the Krt1 gene relative to the expression level of GAPDH. 17B shows the expression level of the Krt5 gene relative to the expression level of GAPDH (Krt5 / GAPDH), FIG. 17C shows the expression level of the Krt14 gene relative to the expression level of GAPDH (Krt14 / GAPDH), FIG. 17D shows the expression level of the Krt16·17 gene relative to the expression level of GAPDH (Krt16·17 / GAPDH), and FIG. 18 shows the expression level of the Eps15 gene relative to the expression level of GAPDH (Eps15 / GAPDH). The results regarding the expression level of the Krt16 gene shown in FIG. 17D can also be interpreted as the results regarding the expression level of the Krt17 gene.

[0125] In Figures 13 to 18, Example AX (0h) shows the results of a hamster euthanized without inducing stomatitis in Test Method 2). Control Example A-1 (3h) and Example AX (3h) show the results of a hamster euthanized 3 hours after inducing stomatitis in Test Method 3). Control Example A-1 (24h) and Example AX (24h) show the results of a hamster euthanized 24 hours after inducing stomatitis in Test Method 3). X is an integer from 3 to 9.

[0126] As shown in FIG. 13, it was confirmed that application of component (A) or component (B) increased the mRNA expression of Pla2g2f gene in the buccal mucosa tissue of hamsters before the induction of stomatitis. As shown in Figures 14 to 18, it was confirmed that application of component (A) or component (B) suppressed the mRNA expression of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene in hamsters with induced stomatitis, compared to control example A-1.

[0127] As a result, the expression of each gene in Test Examples 1 to 3 showed similar trends.

[0128] <Test Example 4: Evaluation of the effect of suppressing the onset of stomatitis> The effects of (A) glycyrrhizinic acid and (B) tranexamic acid on stomatitis, which is thought to be an example of a disease caused by the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, were confirmed.

[0129] (Examples B-1, B-2, and B-3) The liquid samples of Examples B-1, B-2, and B-3 were prepared according to the formulations shown in Table 2, and the following tests were carried out. As shown in Table 2, the amount of dipotassium glycyrrhizinate in Examples B-1 and B-3 was 0.01 w / v%, and the amount of glycyrrhizinic acid was 0.0065 w / v%.

[0130] (Comparative examples B-1, B-2, B-3, B-4) According to the formulations shown in Table 2, liquid samples of Comparative Examples B-1, B-2, B-3 and B-4 were prepared and similarly tested.

[0131] (Control Example B-1) A similar test was performed using saline.

[0132] [Table 2]

[0133] (Test Method) 1) For each group, 2-4 hamsters (Syrian, male, 5 weeks old) that had been pre-bred for one week were given 50μL of the above liquid sample twice a day on the right buccal mucosa of each animal. The first and second applications were performed at an interval of at least 3 hours. The test was performed for six consecutive days. 2) On the 5th and 6th days, in addition to the procedure in 1), a Φ6 mm filter paper soaked in 30 (w / v)% acetic acid aqueous solution was applied to the right buccal mucosa under anesthesia for 10 seconds to induce stomatitis (inflammatory edema). 3) On the seventh day, the presence or absence of inflammatory edema in the right buccal mucosa of each hamster was visually confirmed. Furthermore, the inhibition rate (%) of stomatitis onset in each group was calculated using the following formula (1), and the average value of 2 to 4 animals was calculated. A value of 50% or more was determined to have an inhibitory effect on stomatitis onset. Inhibition rate of stomatitis (%) = (number of animals without inflammatory edema / total number of animals) × 100 (1)

[0134] The results are shown in Table 3.

[0135] [Table 3]

[0136] As shown in Table 3, Examples B-1 and B-2, in which the expression of the Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family and / or Eps15 gene was suppressed in Test Examples 1 and 2, have an excellent effect of suppressing the onset of stomatitis, and it was found that Example B-3, which uses a combination of (A) glycyrrhizinic acid and (B) tranexamic acid, exhibits an even higher effect of suppressing the onset of stomatitis. On the other hand, it was found that comparative examples B-1, B-2, B-3 and B-4, which contain allantoin and sodium azulene sulfonate, known as active ingredients for treating stomatitis, did not have the effect of suppressing the onset of stomatitis. Thus, it was found that the preventive effect against stomatitis is due to the suppression of expression of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and / or the Eps15 gene, and differs from the therapeutic effect after the onset of stomatitis.

[0137] <Test Example 5: Evaluation of the effect of suppressing the onset of stomatitis> The effects of (A) glycyrrhizinic acid and (B) tranexamic acid on stomatitis, which is considered to be an example of a disease caused by Arid5a gene expression, a disease caused by Rac2 gene expression, a disease caused by Clic4 gene expression, a disease caused by keratin gene family expression, and a disease caused by Eps15 gene expression, were confirmed.

[0138] (Example C-1) A liquid sample of Example C-1 was prepared according to the formulation shown in Table 4, and the following tests were carried out. As shown in Table 4, the blending amount of dipotassium glycyrrhizinate was 0.015 w / v%, and the blending amount in terms of glycyrrhizinic acid was 0.00975 w / v%.

[0139] (Comparative example C-1) A liquid sample of Comparative Example C-1 was prepared according to the recipe shown in Table 4 and similarly tested.

[0140] (Control Example C-1) A similar test was performed using saline.

[0141] [Table 4]

[0142] (Test Method) 1) For each group, 5 hamsters (Syrian, male, 5 weeks old) were pre-bred for one week, and the above liquid sample was applied to the right buccal mucosa of each animal twice a day, 50μL each. The first and second applications were performed at an interval of at least 3 hours. The test was performed for six consecutive days. 2) On the 5th and 6th days, in addition to the procedure in 1), a Φ6 mm filter paper soaked in 30 (w / v)% acetic acid aqueous solution was applied to the right buccal mucosa under anesthesia for 10 seconds to induce stomatitis (inflammatory edema). 3) On the seventh day, the presence or absence of inflammatory edema in the right buccal mucosa of each hamster was visually confirmed. Furthermore, the inhibition rate (%) of stomatitis onset in each group was calculated using the following formula (1), and the average of the five animals was calculated. A value of 50% or more was determined to have an inhibitory effect on stomatitis onset. Inhibition rate of stomatitis (%) = (number of animals without inflammatory edema / total number of animals) × 100 (1)

[0143] The results are shown in Table 5.

[0144] [Table 5]

[0145] From Table 5, it was found that Example C-1, which is a composition according to one embodiment of the present invention, has a remarkably high and excellent effect of suppressing the onset of stomatitis. Comparative Example C-1, which does not contain components (A), (B), and (C) in the formulation of Example C-1, showed results similar to those of Control Example C-1, which used saline, and had no effect of suppressing the onset of stomatitis.

[0146] <Test Example 6: Evaluation of therapeutic effect on stomatitis> The liquid samples of Example C-1, Comparative Example C-1 and Control Example C-1 in Test Example 5 were subjected to the following tests.

[0147] (Test Method) 1) For each group, five hamsters (Syrian, male, 5 weeks old) were anesthetized and fixed to a corkboard, and 30 μL of 10 (w / v)% acetic acid saline was injected intradermally into the right cheek pouch to induce stomatitis. 2) After measuring the major and minor axes of the wound and calculating the wound area, 50 μL of the liquid sample was applied to each animal. The liquid sample was applied once a day for six consecutive days. 3) The daily stomatitis healing rate (%) was calculated using the following formula (2) and the average value for the five mice was calculated. Samples were processed from days 1 to 6, but on day 7, no sample processing was performed, and only the major and minor axes of the wound were measured to calculate the wound area. Stomatitis healing rate (%) = {(area at the time of wound creation – area at the time of measurement) / area at the time of wound creation} × 100 (2)

[0148] The results are shown in Table 6.

[0149] [Table 6]

[0150] On the fourth day, the stomatitis healing rate of Example C-1 was 34.3%, which was higher than the stomatitis healing rate of Comparative Example C-1 (9.0%). Even on the seventh day, the stomatitis healing rate of Example C-1 was 49.8%, which was higher than the stomatitis healing rate of Comparative Example C-1 (30.5%).

[0151] Test Examples 5 and 6 show that Example C-1 has a significantly higher effect of suppressing the onset of stomatitis than Comparative Example C-1, and further has a therapeutic effect on stomatitis. This shows that the composition of one embodiment of the present invention has a high preventive and therapeutic effect on stomatitis, which is considered to be an example of a disease caused by Arid5a gene expression, a disease caused by Rac2 gene expression, a disease caused by Clic4 gene expression, a disease caused by keratin gene family expression, and a disease caused by Eps15 gene expression.

[0152] <Test Example 7: Evaluation of the effect of suppressing the onset of stomatitis in an anticancer drug administration model> (Example D-1) A liquid sample of Example D-1 was prepared according to the recipe shown in Table 7, and the following tests were carried out. Note that Example D-1 has the same recipe as Example C-1, with the blending amount of dipotassium glycyrrhizinate being 0.015 w / v% and the blending amount in terms of glycyrrhizinic acid being 0.00975 w / v%.

[0153] (Comparative example D-1) A liquid sample of Comparative Example D-1 was prepared according to the formulation shown in Table 7, and the following tests were carried out.

[0154] (Control Example D-1) The control sample C-1 (physiological saline) used in Test Example 5 was subjected to the same test.

[0155] (Control Example D-2) A similar test was performed without treating the liquid samples.

[0156] [Table 7]

[0157] (Test Method) 1) For each group, 50 μL of the above liquid sample was applied to the buccal mucosa of four anesthetized hamsters (Syrian, male, 6 weeks old) for 10 consecutive days. 2) On the first and second days, the anticancer drug 5-fluorouracil (5FU; 60 mg / kg) was administered intraperitoneally to the hamsters, with a two-day interval. 3) On the fifth day, 20 μL of 5 (w / v)% acetic acid saline was injected into the right cheek pouch of each hamster to induce stomatitis. 4) The wound area was measured from the 6th to the 10th day, and the average value of the 4 animals was recorded as the value for each group. 5) In control example D-2, 4 hamsters (Syrian, male, 6 weeks old) were subjected to 3) and 4) without carrying out 1) and 2) above, and the average wound area was calculated.

[0158] The results are shown in Table 8.

[0159] [Table 8]

[0160] From the control examples D-1 and D-2, it was found that administration of the anticancer drug 5-fluorouracil (5FU) made it easier to develop stomatitis and tended to increase the area of ​​the wound. Comparative example D-1, which contained sodium azulene sulfonate, known as an active ingredient in treating stomatitis, developed stomatitis of the same size as control example D-1 on the sixth day after the onset of stomatitis, and the therapeutic effect of suppressing the inflammation each day was confirmed. In contrast, the wound area of ​​Example D-1, which is a composition according to one embodiment of the present invention, was significantly smaller on the sixth day than that of Comparative Example D-1, demonstrating its excellent effect of suppressing the onset of stomatitis.

[0161] <Reference example: Evaluation of the effect of suppressing and promoting gene expression on stomatitis> As described in detail below, hamsters in which the expression of a specific gene was suppressed were used to observe the effect of suppressing gene expression on the development of stomatitis.

[0162] (Reference Example 1: Establishment of conditions for introducing foreign genes into hamster buccal mucosa by electroporation) First, we performed an experiment to establish the conditions for introducing foreign genes into the buccal mucosa of hamsters by electroporation.

[0163] (Preparation of EGFP gene solution) An EGFP gene solution was prepared in a PBS solution containing 0.1 w / v % trypan blue, so that pCAGGS-EGFP (Enhanced Green Fluorescence Protein) (manufactured by Nepa Gene Co., Ltd.) was at 1 μg / μL.

[0164] (Introduction of EGFP gene) Three hamsters (Syrian, male, 5 weeks old) were fixed to a corkboard under anesthesia, and then 40 μL of the EGFP gene solution was intradermally injected into the right cheek mucosa of each hamster using a syringe (BD Lordose (registered trademark) (30G): manufactured by Becton Dickinson Japan Co., Ltd.). A photograph taken immediately after the intradermal injection is shown in Figure 19A. Figure 19A shows the state in which the injection site on the right cheek mucosa of the hamster was opened so that it could be seen.

[0165] Thereafter, a needle was inserted into the injection site, and the injection site was clamped with a tweezers-type circular platinum electrode (CUY650P5: manufactured by Nepa Gene Co., Ltd.) After adjusting the electrical resistance to about 0.5 KΩ, electroporation was performed under the electrical pulse conditions in Table 9.

[0166] [Table 9]

[0167] The introduction of foreign genes by the electroporation method described above was performed using a Super Electroporator NEPA21 (registered trademark) Type II (manufactured by Nepa Gene Co., Ltd.). This device creates micropores in the cell membrane with a poring pulse (high voltage, short time, multiple times, decay rate setting), and then delivers genes or drugs into the cells multiple times with a transfer pulse (low voltage, long time, multiple times, decay rate setting). Electroporation was performed by clamping the tissue with tweezers electrodes.

[0168] One week after electroporation, the gene transfer site of the anesthetized hamster was irradiated with EGFP excitation wavelength (wavelength around 489 nm) using UV light (Ex Flashlight ExF-B: manufactured by Biotools Co., Ltd.), and the fluorescence wavelength (508 nm) derived from the EGFP protein was confirmed. At this time, protective glasses (Sharp Cut Goggles Yellow: manufactured by Biotools Co., Ltd.) were used.

[0169] Figure 19B shows a photograph of the hamster buccal mucosa emitting the fluorescent wavelength derived from the EGFP protein obtained by the above procedure. In Figure 19B, the hamster buccal mucosa was opened so that the site where the foreign gene was introduced could be seen, and the introduction site was irradiated with UV light. Within the dotted frame, green fluorescence derived from the EGFP protein was macroscopically observed. In all hamsters into which foreign genes had been introduced by electroporation, a specific and strong fluorescent wavelength derived from the EGFP protein was confirmed at the gene introduction site in the buccal mucosa.

[0170] This confirmed that the exogenously introduced gene was stably expressed (functioned) in the recipient.

[0171] The hamsters were then euthanized with carbon dioxide gas. Tissues were collected and fixed in 4% formalin solution, then paraffin blocks were prepared and 4.5 μm-thick slices were prepared using a microtome. Immunohistochemical staining of EGFP protein was performed according to standard methods using the following antibodies (primary antibody: Rabbit Polyclonal antibody to GFP, 1:1000 (#GTX113617, GeneTex), secondary antibody: Envision + System-HRP Labelled Polymer Anti-Rabbit, Dako).

[0172] Figure 20A shows a photograph of a hamster not having the EGFP gene introduced therein, stained with immunohistochemistry for the EGFP protein by the above procedure, and Figure 20B shows a photograph of a site into which the EGFP gene had been introduced, stained with immunohistochemistry by the above procedure. The dotted frame in Figure 20B shows that the EGFP gene had been introduced. This confirmed that the region into which the EGFP gene had been introduced extended over a wide area, from the basal layer to the muscular layer.

[0173] The EGFP protein was histologically confirmed in all hamster buccal mucosa tissues into which foreign genes were introduced, confirming that the foreign genes were introduced locally into the buccal mucosa tissue and were stably expressed. Based on this, we concluded that the conditions for locally introducing foreign genes into the buccal mucosa of hamsters by electroporation have been established.

[0174] (Reference Example 2: Specific gene knockdown test using RNA interference method) Under the conditions established in Reference Example 1, hamsters in which specific genes were knocked down as described below were prepared, and the onset of stomatitis was observed. To knock down specific genes, we used RNA interference, a technique that uses small interfering RNA (siRNA) to destroy messenger RNA (mRNA) in a sequence-specific manner, thereby selectively suppressing (knocking down) the expression of that mRNA.

[0175] (Preparation of siRNA solution) All siRNAs shown below were designed and synthesized by Nippon Gene Co., Ltd. An siRNA solution was prepared so that the siRNA for knocking down the Arid5a gene described below was 100 nM in a PBS solution containing 0.1 w / v% trypan blue. A blank was used as a control, a PBS solution containing 0.1 w / v% trypan blue (siRNA blank). An siRNA solution was prepared in the same manner, using the siRNA for knocking down other genes described below instead of the siRNA for knocking down the Arid5a gene.

[0176] ((siRNA information)) The siRNA information is as follows. The d indicates that the immediately following base is deoxygenated. The underlined parts are DNA, and the rest are RNA.

[0177] [ka]

[0178] (Introduction of siRNA) Five hamsters were used to introduce siRNA for knocking down the Pla2g2f gene, and three hamsters were used to introduce siRNA for knocking down other genes, and the siRNA solution or blank solution was used instead of the EGFP gene solution, but the siRNAs were introduced into the hamsters by electroporation in the same manner as in Reference Example 1. Note that PBS containing 0.1 w / v% trypan blue was used as the blank solution.

[0179] (Induction of stomatitis) 24 hours after electroporation, a 6 mm filter paper soaked in 30% acetic acid solution was applied to the siRNA-injected site of the buccal mucosa of the anesthetized hamster for 10 seconds to induce stomatitis.

[0180] 24 hours after induction of stomatitis, the degree of inflammatory edema (stomatitis) at the site of stomatitis induction was visually observed under anesthesia according to the scores in Table 10. The degree of inflammatory edema (stomatitis) in hamsters with Arid5a gene or Rac2 gene knockdown and control hamsters is shown in Figure 21.

[0181] [Table 10]

[0182] In addition, the Pla2g2f gene knockdown hamster and the control hamster were anesthetized and fixed on a corkboard in a supine position, and then photographed with an iPhone (registered trademark) SE with their mouths open so that the entire stomatitis was included. The area of ​​the stomatitis was measured based on the photographed images using ImageJ, an open source image processing software. The results are shown in Figure 22.

[0183] As shown in Figure 21, the stomatitis score in the blank hamsters was 2.4, whereas the stomatitis score in the hamsters in which the Arid5a gene or Rac2 gene was knocked down by RNA interference was less than 1.5, indicating that the onset of stomatitis was suppressed. Furthermore, as shown in FIG. 22, in hamsters in which the Pla2g2f gene was knocked down, the induced stomatitis tended to worsen compared to the blank-controlled hamsters.

[0184] From the Reference Examples, it is believed that the gene expression of Arid5a or Rac2 contributes to the onset of stomatitis, and the gene expression of Pla2g2f contributes to the inhibition of stomatitis.

Claims

1. An agent for inhibiting the expression of any one gene selected from the group consisting of Arid5a gene, Rac2 gene, Clic4 gene, keratin gene family, and Eps15 gene, comprising at least one of (A) glycyrrhizinic acid and (B) tranexamic acid.

2. A composition for inhibiting the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, comprising at least one of (A) glycyrrhizinic acid and (B) tranexamic acid as active ingredients.

3. A composition for at least one of preventing and treating a disease caused by expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family and the Eps15 gene, comprising the gene expression inhibitor of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family and the Eps15 gene described in claim 1.

4. The composition according to claim 3, which is for at least one of preventing and treating a disease caused by the expression of any one gene selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, is stomatitis.

5. An agent for inhibiting the expression of two or more genes selected from the group consisting of the Arid5a gene, the Rac2 gene, the Clic4 gene, the keratin gene family, and the Eps15 gene, comprising at least one of (A) glycyrrhizinic acid and (B) tranexamic acid.

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