Inhibin βA (INHBA) expression inhibitor

Abstract

To discover a substance that can suppress the expression of activin A, and provide the discovered substance as an agent for treating and / or preventing diseases or pathological conditions associated with the enhanced expression (overexpression) of activin A.SOLUTION: An INHBA expression inhibitor contains rice extract, or a mixture of rice extract and purple wheat seed extract.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an inhibitor of inhibin βA (INHBA) expression. [Background technology]

[0002] Inhibin βA (INHBA) is highly conserved among vertebrates, and after extracellular secretion, it dimerizes to become activin A. Therefore, it can be said that the quantitative change of INHBA in the body is positively correlated with the quantitative change of activin A. Activin A was identified as a gonadal protein that induces the biosynthesis and secretion of follicle-stimulating hormone from the anterior pituitary gland (Non-Patent Document 1). Previous studies have confirmed that INHBA / activin A is expressed in various tissues from embryonic development to adulthood, and has been shown to be involved in various biological functions, such as stem cell maintenance and differentiation, cell proliferation, apoptosis, immune response, and inflammatory response (Non-Patent Document 2).

[0003] Activin A transmits signals via two receptors, type I and type II receptors. When activin A binds to the type II receptor, the type I receptor is recruited, forming a tetrameric receptor complex and undergoing phosphorylation. The activated type I receptor phosphorylates intracellular SMAD2 / 3, which then translocates to the nucleus and regulates target gene transcription (Non-Patent Documents 2 and 3). SMAD-independent signal transduction is also possible. Activated type I receptors activate intracellular signaling pathways, such as extracellular signal-regulated kinases 1 / 2 (ERK 1 / 2), p38 mitogen activated protein kinase (MAPK), and c-Jun N-terminal kinases (JNK), affecting cell migration and differentiation (Non-Patent Document 4). Activin A is also known to induce the Wnt signaling pathway by activating SMAD2 (Non-Patent Document 5).

[0004] As described above, activin A is involved in multiple intracellular signaling mechanisms and regulates diverse cellular responses, making it essential for maintaining the function of cells that constitute biological tissues. On the other hand, due to its involvement in various biological actions and intracellular signaling mechanisms, its abnormal expression is associated with various diseases and pathologies. In fact, activin A is overexpressed in tumor cells of lung adenocarcinoma, esophageal cancer, and malignant pleural mesothelioma, promoting cell proliferation and tumor expansion. It has also been reported that activin A promotes tumor cell invasion and metastasis in several tumors, including prostate cancer, breast cancer, oral squamous cell carcinoma, and melanoma (Non-Patent Documents 6-8). In addition to cancer, increased activin A has also been reported to be involved in autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis, allergic diseases such as allergic asthma and atopic dermatitis, and bone metabolic disorders (Non-Patent Documents 9 and 10).

[0005] Therefore, the development of treatments that control activin A signaling for such diseases is underway. For example, by intraperitoneally injecting follistatin, an activin A antagonist protein, into colitis model mice, preventing activin A receptor binding and initiation of the signaling pathway, it has been observed that colon inflammation is suppressed and survival rates are increased (Non-Patent Document 11). In addition, there are methods that use anti-activin A antibodies or activin antagonists to inhibit activin A signaling and treat diseases and disorders (Patent Documents 1 to 4).

[0006] Therefore, appropriate control of activin A signaling is useful not only for the treatment of various diseases but also for efficiently controlling stem cell differentiation and cell proliferation. However, the use of antagonistic proteins and antibody proteins such as those mentioned above is problematic because they are easily degraded by enzymes present in the body and also bind to transforming growth factor-β (TGF-β) and myostatin (GDF-8), which are structurally similar to activin A. Therefore, the elucidation of new factors to replace these is desired. [Prior art documents] [Patent documents]

[0007] [License 1] Special Announcement No. 2020-11965 [License 2] Special Announcement No. 2021-512919 [License 3] Special Announcement No. 2018-162312 [License 4] Special Announcement No. 2018-145198 [Non-licensed literature]

[0008] [Non-licensed Document 1] Ling N, Ying SY, Ueno N, et al. Pituitary FSH is released by a heterodimer of the beta-subunits from the two forms of inhibin. Nature. 1986;321(6072):779-782. [Non-licensed Document 2] Morianos I, Papadopoulou G, Semitekolou M, Xanthou G. Activin-A in the regulation of immunity in health and disease. J Autoimmun. 2019;104:102314. [Non-licensed Document 3] Namwanje M, Brown CW. Activins and inhibins: roles in development, physiology, and disease. Cold Spring Harb Perspect Biol. 2016;8(7):a021881. [Non-licensed Document 4] de Guise C, Lacerte A, Rafiei S, et al. Activin inhibits the human Pit-1 gene promoter through the p38 kinase pathway in a Smad-independent manner. Endocrinology. 2006;147(9):4351-4362.

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[0009] Therefore, the objective of the present invention is to find a substance that can suppress the expression of activin A, which controls various functions such as cell proliferation and differentiation, apoptosis induction, immune response, and inflammatory response, and to provide this as a drug for treating and / or preventing diseases or pathological conditions associated with increased expression (overexpression) of activin A. [Means for solving the problem]

[0010] As a result of intensive research to solve the above-mentioned problems, the inventors discovered that rice extract, or a mixture of rice extract and purple barley seed extract, significantly suppresses the expression of inhibin βA (INHBA), a subunit of activin A, and thus completed the present invention.

[0011] That is, the present invention includes the following inventions. (1) An inhibin βA (INHBA) expression inhibitor containing rice extract as an active ingredient. (2) An inhibin βA (INHBA) expression inhibitor containing a mixture of rice extract and purple barley seed extract. (3) An inhibin βA (INHBA) expression inhibitor according to (1) or (2), characterized in that the extract is extracted with one or more solvents selected from the group consisting of water, lower alcohols, and hydrocarbons. (4) An inhibin βA (INHBA) expression inhibitor according to (1) or (2), which is used for the treatment and / or prevention of a disease or pathology associated with increased expression of activin A. [Effects of the Invention]

[0012] The INHBA expression inhibitor of the present invention can significantly suppress the expression of INHBA in cells, and is therefore effective in treating and / or preventing diseases and pathological conditions associated with increased expression of activin A, an INHBA homodimer. DETAILED DESCRIPTION OF THE INVENTION

[0013] The inhibin βA (INHBA; hereinafter referred to as "INHBA") expression inhibitor of the present invention contains, as an active ingredient, a rice extract or a mixture of a rice extract and a purple barley seed extract. In the present invention, "inhibition of INHBA expression" refers to the inhibition of INHBA expression at the living organism level or culture level. Furthermore, in the present invention, "inhibition of INHBA expression" refers to the inhibition of INHBA mRNA expression and protein expression.

[0014] INHBA and its homodimer, activin A, are proteins belonging to the transforming growth factor-β (TGF-β) family and regulate many physiological functions, including cell proliferation and differentiation, apoptosis induction, immune responses, and inflammatory responses, by controlling the activin A signaling pathway. The sequence information for the INHBA gene and protein, for example, for humans, is registered as GenBank numbers Nucleotide NM_002192.4 and Protein NP_002183.1.

[0015] The rice used in the present invention is the seed of Oryza sativa L., which belongs to the Poaceae family, and any of brown rice (rice obtained by removing the husk from unhulled rice), germinated rice (rice obtained by germinating brown rice), germinated rice (rice obtained by removing the bran from brown rice and leaving the germ), polished rice (rice obtained by removing the bran and germ from brown rice, leaving only the endosperm), rice bran, germ, and endosperm may be used. The variety is not particularly limited, and examples include edible varieties such as Koshihikari, Sasanishiki, and Akitakomachi.

[0016] When brown rice or polished rice is used, it may be subjected to a pulverization treatment, for example, using a ball mill, a roll mill, an impact mill, an ultracentrifugal mill, an airflow mill, or a shear friction mill.

[0017] Purple barley used in the present invention refers to barley with purple glumes, caryopses, stems, or leaves, and its scientific name is Hordeum vulgare L. The variety is not particularly limited, but examples include OUC321, CI158, and CI244, which can be obtained from the Wild Barley Resources Research Center, Institute of Bioresources, Okayama University.

[0018] The extraction method for obtaining rice extracts and purple barley seed extracts is not particularly limited, and may be, for example, a heated extraction method, or a room temperature or cold extraction method. Examples of solvents used for extraction include water, lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), hydrocarbons (hexane, heptane, liquid paraffin, squalane, etc.), liquid polyhydric alcohols (1,3-butylene glycol, propylene glycol, glycerin, etc.), ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), and ethers (ethyl ether, tetrahydrofuran, propyl ether, etc.). Among these solvents, water, lower alcohols, and hydrocarbons are preferred, with lower alcohols and hydrocarbons being more preferred, and hydrocarbons being even more preferred. Particularly preferred extraction solvents include ethanol and hexane. These solvents may be used alone or in combination. Furthermore, among the above solvents, the water-soluble organic solvent may be used alone, or a mixed solvent of water and a water-soluble organic solvent (so-called aqueous organic solvent) may be used. The concentration of the organic solvent contained in the aqueous organic solvent is not limited, but for example, in the case of aqueous ethanol (ethanol aqueous solution), it is 10 to 90 v / v%, preferably 30 to 70 v / v%. In addition, a solvent whose pH has been adjusted by adding an acid or alkali to the above extraction solvent may also be used.

[0019] The amount of solvent used is not particularly limited, and may be, for example, 10 times or more, preferably 20 times or more, relative to the amount of rice or purple barley seeds (dry weight). However, for convenience of operations such as concentration and isolation after extraction, it is preferable that the amount be 100 times or less. The extraction temperature and time depend on the type of solvent used, but examples include 10 to 100°C, preferably 30 to 90°C, and 30 minutes to 24 hours, preferably 1 to 10 hours. More specifically, a rice or purple barley seed extract can be obtained by adding water to dried rice or purple barley seeds and performing hot water extraction at 95 to 100°C. Alternatively, a rice or purple barley seed extract can be obtained by adding a lower alcohol (e.g., ethanol), a liquid polyhydric alcohol (e.g., propylene glycol, 1,3-butylene glycol), and / or a hydrocarbon (hexane, heptane, liquid paraffin, squalane, etc.) to dried rice or purple barley seeds and performing extraction at room temperature (e.g., 15 to 35°C). The extract may be used as the extracted solution as is, or, if necessary, may be subjected to treatments such as concentration (e.g., by organic solvents, vacuum concentration, membrane concentration, etc.), dilution, filtration, decolorization with activated carbon, deodorization, ethanol precipitation, etc., within a range that does not affect the extract's effectiveness. Furthermore, the extracted solution may be concentrated to dryness, spray-dried, freeze-dried, etc., and used as a dried product.

[0020] In the present invention, rice extract may be used alone, but it is preferable to use it in combination with purple barley seed extract, as this enhances the INHBA expression-suppressing effect. When rice extract and purple barley seed extract are used in combination, the mixing ratio is not limited, but is preferably 1:10 to 10:1, more preferably 1:5 to 5:1, even more preferably 1:2 to 2:1, and most preferably 1:1.

[0021] Cells targeted for INHBA expression inhibition are not particularly limited as long as they express INHBA and are present in biological tissues. Examples include cells from bone marrow, brain, head and neck, nasopharynx, bone, blood, skin, heart, lung, breast, esophagus, stomach, pancreas, liver, colon and rectum, small intestine, spleen, testis, ovary, thyroid, bladder, salivary gland, duodenum, endometrium, placenta, prostate, gallbladder, kidney, and lymph node. The cells may also be undifferentiated pluripotent stem cells, such as embryonic stem cells (ES cells); somatic stem cells present in bone marrow, blood, skin (epidermis, dermis, and subcutaneous tissue), fat, hair follicles, brain, nerves, liver, pancreas, kidney, muscle, and other tissues; and stem cells artificially generated by gene transfer (induced pluripotent stem cells: iPS cells). Cells of mammalian origin, such as human, mouse, rat, hamster, rabbit, dog, cat, pig, cow, horse, and monkey, are preferred. The cells may be in vivo cells or isolated cells, which can be maintained and cultured by known techniques.

[0022] Rice extract or a mixture of rice extract and purple barley seed extract, by virtue of the inhibitory effect of INHBA on expression, is effective in treating and / or preventing various diseases or conditions associated with increased expression (overexpression) of activin A. Here, diseases or conditions associated with increased expression of activin A include cancer (colon cancer, melanoma, gastric cancer, hepatocellular carcinoma, prostate cancer, esophageal cancer, pancreatic cancer, lung cancer, breast cancer, kidney cancer, bladder cancer, cervical cancer, ovarian cancer, thyroid cancer, head and neck cancer, lymphoma, glioma, glioblastoma, etc.), allergic diseases (hay fever, atopic dermatitis, bronchial asthma, allergic rhinitis (runny nose, nasal congestion, sneezing), allergic conjunctivitis, allergic gastroenteritis, bronchial asthma, food allergies), and the like. , urticaria, collagen disease, serum sickness, viral hepatitis, contact dermatitis, etc.), autoimmune diseases (rheumatoid arthritis, multiple sclerosis, systemic lupus erythematosus, Sjogren's syndrome, type 1 diabetes, polymyositis, systemic sclerosis, dermatomyositis, pernicious anemia, etc.), inflammatory bowel disease (ulcerative colitis, Crohn's disease, etc.), nervous system diseases accompanied by nerve degeneration or damage (memory loss, dementia, Alzheimer's disease, spinal cord injury, facial nerve paralysis, spinocerebellar degeneration, Huntington's disease (HD), amyotrophic lateral sclerosis, etc.), Amyotrophic lateral sclerosis (ALS), polyneuropathy, spinal muscular atrophy, Parkinson's disease, multiple sclerosis (MS), cerebrovascular disorders (cerebral infarction, stroke, cerebral aneurysm), movement disorders due to cerebrovascular disorders, progressive supranuclear palsy, tremors, epilepsy, brain trauma, depression, insomnia, learning disabilities, anxiety disorders (panic disorder, obsessive-compulsive disorder, etc.), muscle diseases (sarcopenia, muscular atrophy, muscular dystrophy, etc.), skin aging and damage (wrinkles, sagging, dullness, loss of firmness and luster, rough skin, dryness) These include, but are not limited to, skin, sensitive skin, hyperkeratosis, enlarged pores, cracks, chapped skin, asteatosis (xeroderma), asteatotic eczema, pruritus, hand eczema, psoriasis (accompanied by erythema, scaling, and desquamation), delayed healing of burns and wounds, bedsores, etc., bone metabolic disorders (osteoporosis, osteomalacia, osteogenesis imperfecta, osteopetrosis, Paget's disease of bone, etc.), infectious diseases (infections caused by influenza virus, norovirus, herpes virus, AIDS virus, etc.).

[0023] The INHBA expression inhibitor of the present invention can be used as is, but can also be provided by incorporating it into various compositions, such as pharmaceuticals, quasi-drugs, cosmetics, foods and beverages, for suppressing INHBA expression, together with appropriate additives within a range that does not impair the effects of the present invention, for the purpose of treating and / or preventing various diseases or pathological conditions associated with the increased expression of activin A. The pharmaceuticals of the present invention also include drugs for use on animals, i.e., veterinary drugs.

[0024] The pharmaceutical of the present invention functions as a preventive drug that suppresses the onset of the above-mentioned diseases or pathological conditions and / or as a therapeutic drug that improves the condition to a normal state. Since the active ingredient of the pharmaceutical of the present invention is derived from a natural product, it is extremely safe and has no side effects, and therefore, when used as a pharmaceutical for treating and / or preventing the above-mentioned diseases or pathological conditions, it can be administered orally or parenterally in a wide range of dosages to mammals such as humans, mice, rats, rabbits, dogs, and cats.

[0025] When the INHBA expression inhibitors of the present invention are incorporated into pharmaceuticals, they can be mixed with pharmacologically and pharmaceutically acceptable additives and formulated into various formulations suitable for application to affected areas. Pharmacologically and pharmaceutically acceptable additives include formulation bases, carriers, excipients, diluents, binders, lubricants, coating agents, disintegrants or disintegration aids, stabilizers, preservatives, antiseptics, bulking agents, dispersants, wetting agents, buffers, solubilizers or solubilizers, isotonicity agents, pH adjusters, propellants, colorants, sweeteners, flavoring agents, and flavoring agents, and can be added as appropriate depending on the dosage form and intended use. Various formulations suitable for oral or parenteral systemic or local administration can be prepared by various known methods. When providing the pharmaceuticals of the present invention in the above forms, they can be manufactured by methods commonly used by those skilled in the art, such as the methods set forth in the General Provisions for Preparations [2] of the Japanese Pharmacopoeia.

[0026] For oral administration formulations, for example, excipients such as starch, glucose, sucrose, fructose, lactose, sorbitol, mannitol, crystalline cellulose, magnesium carbonate, magnesium oxide, calcium phosphate, or dextrin; disintegrants or disintegration aids such as carboxymethylcellulose, carboxymethylcellulose calcium, starch, or hydroxypropylcellulose; binders such as hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, gum arabic, or gelatin; lubricants such as magnesium stearate, calcium stearate, or talc; coating agents such as hydroxypropylmethylcellulose, sucrose, polyethylene glycol, or titanium oxide; and bases such as petrolatum, liquid paraffin, polyethylene glycol, gelatin, kaolin, glycerin, purified water, or hard fat can be used, but are not limited to these.

[0027] Formulations for parenteral administration may contain, but are not limited to, solvents such as distilled water, saline, ethanol, glycerin, propylene glycol, macrogol, alum water, and vegetable oil; isotonicity agents such as glucose, sodium chloride, and D-mannitol; and pH adjusters such as inorganic acids, organic acids, inorganic bases, and organic bases.

[0028] The pharmaceutical form is not particularly limited, and examples thereof include oral preparations such as tablets, sugar-coated tablets, capsules, lozenges, granules, powders, liquids, pills, emulsions, syrups, suspensions, and elixirs; and parenteral preparations such as injections (e.g., subcutaneous injections, intravenous injections, intramuscular injections, and intraperitoneal injections), drip infusions, suppositories, ointments, lotions, eye drops, sprays, transdermal absorption preparations, transmucosal absorption preparations, and patches. The pharmaceutical may also be prepared as a dried product that is reconstituted before use, and injectable preparations are provided in unit-dose ampoules or multi-dose containers. Furthermore, when used as a pharmaceutical for treating, ameliorating, or preventing skin-related diseases or symptoms associated with increased expression of activin A, the pharmaceutical form suitable for use is an external preparation, such as an ointment, cream, gel, liquid, patch (cataplasm, plaster), foam, spray, or aerosol. Ointments are homogeneous, semi-solid topical preparations, including oil-based ointments, emulsion-based ointments, and water-soluble ointments. Gels are topical preparations in which a water-insoluble component, a hydrated compound, is suspended in an aqueous liquid. Solutions are liquid topical preparations, including lotions, suspensions, emulsions, and liniments.

[0029] When the INHBA expression inhibitor of the present invention is incorporated into cosmetics or quasi-drugs, the formulation may be any of an aqueous solution, solubilized solution, emulsion, powder, powder dispersion, oil solution, gel, ointment, aerosol, water-oil two-layer system, or water-oil-powder three-layer system. Furthermore, such cosmetics or quasi-drugs can be produced by appropriately blending various ingredients, additives, bases, etc. commonly used in skin topical compositions, selected according to their types, together with the INHBA expression inhibitor of the present invention, according to methods known in the art. The formulation may be any of a liquid, emulsion, cream, gel, paste, spray, etc. The ingredients include, for example, oils and fats (olive oil, coconut oil, evening primrose oil, jojoba oil, castor oil, hardened castor oil, etc.), waxes (lanolin, beeswax, carnauba wax, etc.), hydrocarbons (liquid paraffin, squalene, squalane, petrolatum, etc.), fatty acids (lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, etc.), higher alcohols (myristyl alcohol, cetanol, cetostearyl alcohol, stearyl alcohol, behenyl alcohol, etc.), esters (isopropyl myristate, isopropyl palmitate ... These include: isopropyl, cetyl octanoate, glycerin trioctanoate, octyldodecyl myristate, octyl stearate, stearyl stearate, etc.), organic acids (citric acid, lactic acid, α-hydroxyacetic acid, pyrrolidone carboxylic acid, etc.), sugars (maltitol, sorbitol, xylobiose, N-acetyl-D-glucosamine, etc.), proteins and protein hydrolysates, amino acids and their salts, vitamins, plant and animal extracts, various surfactants, moisturizers, UV absorbers, antioxidants, stabilizers, preservatives, disinfectants, fragrances, etc.

[0030] Examples of types of cosmetics and quasi-drugs include lotions, emulsions, gels, beauty serums, general creams, sunscreen creams, packs, masks, facial cleansers, cosmetic soaps, foundations, powders, and body lotions.

[0031] When the extract is incorporated into the above-mentioned pharmaceuticals, quasi-drugs, and cosmetics, its content is not particularly limited, but is preferably 0.001 to 30 wt% (w / w), and more preferably 0.01 to 10 wt% (w / w), calculated as the dry solid content of the extract relative to the total weight of the formulation (composition). At less than 0.001 wt% (w / w), the effect is low, and at more than 30 wt% (w / w), the effect is unlikely to be significantly enhanced. Furthermore, the active ingredient may be added in advance or during production, depending on the ease of use.

[0032] The INHBA expression inhibitor of the present invention can also be incorporated into food and beverage products. In the present invention, the term "food and beverage product" refers not only to general food and beverage products but also to foods other than pharmaceuticals that can be consumed for the purpose of maintaining or improving health, such as health foods, functional foods, health-promoting foods, or foods for special dietary uses. Health foods include foods offered under names such as dietary supplements, dietary supplements, and supplements. Health-promoting foods are defined by the Food Sanitation Act or the Health Promotion Act and include foods for specified health uses and foods with nutrient functions, which can display specific health benefits, nutritional component functions, and disease risk reduction, as well as foods with functional claims, which can display scientifically based functionality reported to the Commissioner of the Consumer Affairs Agency. Special dietary foods also include foods for patients, the elderly, infants, and pregnant women, which display information indicating that the foods are suitable for specific individuals or patients with specific diseases. The specific health benefits and nutritional component functions of foods and beverage products can be displayed on product containers, packaging, instructions, package inserts, and other labels, as well as in product flyers, pamphlets, and product advertisements in newspapers and magazines.

[0033] The form of the food or drink may be any form suitable for consumption, such as solid, liquid, granule, powder, capsule, cream, or paste. In particular, in the case of the above-mentioned health foods, etc., preferred forms include tablet, pill, capsule, powder, granule, fine granule, lozenge, liquid (including syrup, milk, and suspension), etc.

[0034] Examples of foods and beverages include, but are not limited to, bread, noodles, confectionery, dairy products, processed seafood and livestock foods, oils and fats and processed foods, seasonings, various beverages (soft drinks, carbonated drinks, beauty drinks, nutritional drinks, fruit drinks, dairy drinks, etc.), and concentrated liquids and powders for adjusting such beverages. Food and beverages may contain additives that are commonly used depending on the type of food and beverage. Any additives permitted under the Food Sanitation Act can be used, including, for example, sweeteners such as glucose, sucrose, fructose, isomerized liquid sugar, aspartame, and stevia; acidulants such as citric acid, malic acid, and tartaric acid; excipients such as dextrin and starch; binders, diluents, flavorings, colorants, buffers, thickeners, gelling agents, stabilizers, preservatives, emulsifiers, dispersants, suspending agents, and antiseptics. [Example]

[0035] The present invention will be explained in more detail below with reference to examples, but the present invention is not limited to these examples.

[0036] [Example 1] Production of rice and purple barley seed extracts Rice (form: rice bran, brown rice, polished rice, variety: "Koshihikari") and purple barley (variety: "OUC321") seeds were used as extraction materials, and extracts thereof were produced by the following method.

[0037] (Production Example 1) Production of hot water extracts of each extraction material 200 mL of purified water was added to 10 g of dried extract material, and the mixture was extracted at 95-100°C for 2 hours, filtered, and the filtrate was concentrated and freeze-dried to obtain a hot water extract.

[0038] (Production Example 2) Production of ethanol extracts of each extraction material 200 mL of ethanol was added to 10 g of dried extract material, and the mixture was extracted at room temperature for 7 days. After filtration, the filtrate was concentrated to dryness to obtain the ethanol extract.

[0039] (Production Example 3) Production of hexane extracts of each extraction material 200 mL of hexane was added to 10 g of dried extract material, and the mixture was extracted at room temperature for 7 days. After filtration, the filtrate was concentrated to dryness to obtain the hexane extract.

[0040] For the extracts obtained in Production Examples 1 to 3 (Extracts 1 to 12), the extraction materials, extraction solvents, and amounts of extract used are shown in Table 1 below.

[0041] [Table 1]

[0042] [Example 2] Inhibitory effect on INHBA expression The inhibitory effect of rice and purple barley seed extracts on INHBA expression was tested as follows.

[0043] In this example, to verify the INHBA-inhibitory effect of each extract, a test was carried out using normal human epidermal keratinocytes (HEK, manufactured by Kurabo Industries, Ltd.).

[0044] First, we investigated ultraviolet (UVB) irradiation to create a model in which INHBA is overexpressed in HEK cells. Specifically, HEK cells were cultured in a 96-well plate at 3 × 10 cells per well. 3 After seeding and confirming the survival of the seeds, the cells were exposed to ultraviolet light (UVB) at 100 mJ / cm using a Toshiba FL20S·E lamp. 2 Two days after UVB irradiation, the cells were fixed with PFA and treated with an antibody against INHBA (Proteintech) and a fluorescently labeled secondary antibody for immunostaining. The nuclei of individual cells were labeled with DAPI staining. The INHBA expression level per cell (INHBA fluorescence intensity / cell number) was then calculated using a fluorescence microscope and image analysis software (ImageJ). The INHBA expression level in cells cultured without UVB irradiation was set at 100%, and the relative INHBA expression level in cells cultured under UVB irradiation was calculated. The results are shown in Table 2.

[0045] [Table 2]

[0046] As shown in Table 2, it was found that UVB irradiation enhanced the expression of INHBA.

[0047] Next, an addition test of each extract was carried out. Specifically, HEK cells were placed in a 96-well plate at 3 × 10 cells per well in the same manner as above. 3 After sowing the seeds individually and confirming their survival, they were exposed to ultraviolet light (UVB) at 100 mJ / cm 2 The cells were irradiated so that the concentration of each extract obtained in Example 1 was 100 μg / mL. When a mixture of two extracts was used, the ratio was 1:1 (by weight). Two days after the addition of the extracts, the cells were fixed with PFA, and the INHBA expression level (INHBA fluorescence intensity / cell count) was calculated in the same manner as above. The INHBA expression level in cells cultured without the addition of the extract (control) was set to 100%, and the relative INHBA expression level in cells cultured with the addition of the extract was calculated and evaluated. The results are shown in Table 3.

[0048] [Table 3]

[0049] As shown in Table 3, in cells cultured in medium supplemented with extracts (extracts 1-9) of rice (rice bran, brown rice, polished rice), the expression level of INHBA was reduced compared to the control (no addition), demonstrating the INHBA expression-suppressing effect. Furthermore, it was confirmed that the combined use of rice extract (extract 6) and purple barley seed extract (extract 12) significantly enhanced the INHBA expression-suppressing effect. [Industrial Applicability]

[0050] The present invention can be used in the field of manufacturing pharmaceuticals and cosmetics for the purpose of treating and / or preventing diseases associated with increased expression of activin A.

Claims

1. An inhibin βA (INHBA) expression inhibitor containing a mixture of rice extract and purple barley seed extract, characterized in that the rice extract and purple barley seed extract are extracted with water or hot water, an aqueous ethanol solution, ethanol, or hexane.

2. The inhibin βA (INHBA) expression inhibitor according to claim 1, which is used for the treatment and / or prevention of a disease or pathological condition associated with increased expression of activin A.

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