JP2023086263A - Collagen production promoter, mmp inhibitor, hyaluronic acid production promoter, cell proliferation promoter, and internal agent - Google Patents

Abstract

To provide novel external or internal agents having great actions of promoting collagen production, inhibiting MMP, promoting hyaluronic acid production and promoting cell proliferation.SOLUTION: It has been found out that extract from germinated purple barley seeds has great actions of promoting collagen production, inhibiting MMP, promoting hyaluronic acid production and promoting cell proliferation and also has high stability. The extract from germinated purple barley seeds can be used in the beauty field such as prevention of skin aging as well as in the medical field such as inhibition of aging-related hypofunction, prevention and treatment of cancer, and is expected to be applied to cosmetic products, food, quasi drugs and pharmaceuticals or the like.SELECTED DRAWING: None

Description

TECHNICAL FIELD The present invention relates to collagen production promoters, MMP inhibitors, hyaluronic acid production promoters, cell proliferation promoters and internal preparations.

Fibroblasts and collagen are present in the dermis, and type I collagen accounts for 80% of the total. In addition to type I collagen, the existence of types III, V, XII and XIV collagen is known. One of the causes of wrinkles and sagging is a decrease in type I collagen. Therefore, promoting the production of type I collagen is considered to be effective in preventing and improving wrinkles and sagging. In addition, promoting the production of type I collagen is also effective in improving skin wound healing.

In addition to UV rays, the skin is exposed to various physical and chemical stresses such as dryness, cold, heat and drugs on a daily basis. As a result, functional deterioration of the skin is caused, and various skin aging phenomena become apparent. One of the skin aging phenomena is wrinkles. It is known that there are two types of wrinkles: epidermal wrinkles and dermal wrinkles. Epidermal wrinkles are called fine wrinkles, and are wrinkles that are temporarily caused by a decrease in the water content in the stratum corneum due to dryness of the skin. On the other hand, dermal wrinkles are wrinkles formed by ultraviolet rays contained in sunlight and aging. Mechanisms for its formation include a decrease in the ability to synthesize collagen in dermal fibroblasts due to ultraviolet light and aging, and an increase in matrix metalloproteinase (MMP) to promote the degradation of collagen.

Epidermal wrinkles caused by dryness and dermal wrinkles differ in histological morphology, onset mechanisms, and treatment methods. Dermal wrinkles caused by UV rays and aging can be improved by using cosmetics with moisturizing effects. is difficult.

So far, for the purpose of improving dermal wrinkles caused by ultraviolet rays, a skin wrinkle formation preventive/improving agent containing hydrolyzed almonds as an active ingredient (Patent Document 1), extracts of mulberry, tenki and marigold has been reported as an active ingredient for improving wrinkles caused by ultraviolet irradiation (Patent Document 2).

MMPs play a major role in infiltration of cancer cells into the interstitium, invasion into blood vessels, and angiogenesis. The interstitium is mainly composed of type I collagen, and the migration of cancer cells requires destruction of the matrix by interstitial collagenase or the like. For the completion of metastasis, it is necessary to destroy the vascular endothelial basement membrane and migrate in the interstitium, and MMPs are also involved in this stage (Non-Patent Document 1). Therefore, a substance having an inhibitory activity against MMP is expected to have an effect of suppressing angiogenesis in cancer tissues and metastasis of cancer, and is considered useful for prevention and treatment of cancer diseases. Thus, MMP inhibition is useful for the prevention, treatment and improvement of various diseases caused by MMP enhancement, such as cancer, ulceration, arteriosclerosis, rheumatoid arthritis, osteoporosis and periodontitis.

Collagenase (MMP-1), which belongs to MMP, is an enzyme produced by fibroblasts, chondrocytes, and the like, and is greatly involved in promoting the degradation of collagen. Collagen is the major structural protein that accounts for approximately one-third of mammalian tissue and is an essential component of many matrix tissues such as cartilage, bone, tendon, gums, and skin. Collagen molecules, which are stable in normal tissue, are denatured into single-stranded gelatin and degraded by a variety of other proteases when cleaved at one point by collagenase. As a result, the structural integrity of the matrix tissue is lost, causing wrinkles, cancer diseases, ulceration, osteoporosis, periodontitis and the like.

As materials having collagenase inhibitory activity, for example, a cacao husk extract (Patent Document 3), which is a cocoa bean skin, a Rosaceae extract (Patent Document 4), lactoferrin (Patent Document 5), and the like have been proposed. Under the circumstances of increasing interest in skin aging and oral hygiene, it is desired to find a highly safe material with no side effects and excellent collagenase activity inhibitory action.

Gelatinase (MMP-2) belonging to MMP is an enzyme produced by fibroblasts, endothelial cells, cancer cells, etc., and is a structural protein that forms a special component of elastic tissues such as collagen, gelatin, and elastin (arteries, tendons, skin, etc.). ) and other substrates. Therefore, when elastin is degraded by gelatinase, the risk of diseases such as cancer, arteriosclerosis, rheumatoid arthritis, and injuries such as ligament rupture increases.

In addition, fibroblasts produce proteins such as collagen and glycosaminoglycans such as hyaluronic acid to form dermal connective tissue and maintain skin firmness. It is thought that wrinkles and sagging of the skin occur as a result of this connective tissue losing contractile force and further losing elasticity.

In particular, hyaluronic acid is known as a high-molecular-weight polysaccharide that is widely distributed in connective tissue, exhibits a gel-like form in the dermis, and maintains skin elasticity. Therefore, it is believed that alteration or reduction of hyaluronic acid is important in skin aging. In addition, since hyaluronic acid is a polymer, there is a problem that cosmetics containing hyaluronic acid are not easily absorbed even when directly applied to the skin. Therefore, an external preparation for skin that can promote the cells' own production of collagen and hyaluronic acid by activating fibroblasts has been sought (Patent Document 6).

Moreover, hyaluronic acid is also present in joints, and is known to function to soften the impact of joint loads and to smooth joint movements. The concentration of hyaluronic acid in normal human joint fluid is about 2.3 mg/mL, but in the case of rheumatoid arthritis, the concentration of hyaluronic acid in synovial fluid decreases to about 1.2 mg/mL, and at the same time the viscosity of synovial fluid increases. significantly reduced (Non-Patent Document 2). In addition, it is known that pyogenic arthritis, gouty arthritis, etc. also cause a decrease in hyaluronic acid content, as in the case of chronic rheumatoid arthritis (Non-Patent Document 3). In the above diseases, increasing the amount of hyaluronic acid in synovial fluid is considered to improve lubricating function, cover and protect articular cartilage, suppress pain, and improve pathological synovial fluid. For example, it is known that joint injection of sodium hyaluronate to rheumatoid arthritis patients improves the above-mentioned symptoms (Non-Patent Document 4). However, treatment of the above diseases is long-term. Therefore, external skin preparations, foods, and pharmaceuticals containing hyaluronic acid production promoters are desired for easy prevention and treatment in daily life.

Floaters are a condition in which faint shadows that look like lint or mosquitoes appear in the field of vision, and are caused by opacities in the vitreous that fill the inside of the eye and cast shadows on the retina. Floaters can be roughly divided into two types: physiological floaters that develop due to aging, ultraviolet rays, active oxygen, etc., and diseases such as retinal detachment, retinal tears, vitreous hemorrhage, and uveitis. There is pathological floaters presenting as one symptom. Physiological floaters are caused by liquefaction due to a decrease in hyaluronic acid, which is a major component of the vitreous, and turbidity in the vitreous due to the accompanying degradation of collagen fibers. Vitrectomy and laser therapy are available as treatment methods, but these procedures are not often performed in Japan due to safety concerns, and treatment overseas requires a large amount of money. . Therefore, in order to prevent and improve physiological floaters, foods and pharmaceuticals containing hyaluronic acid production promoters that can be used on a daily basis are desired.

In general, with aging, the proliferation/division ability of epidermal keratinocytes decreases, and the epidermal layer itself becomes thinner (Non-Patent Document 5). Biological factors such as epidermal growth factor (EGF/epidermal growth factor) and female hormones (estrogen) act on epidermal keratinocyte proliferation in the skin, but their secretion decreases with aging. Such deterioration in epidermal keratinocyte metabolic function due to aging delays the turnover rate of the skin and causes rough skin and aging of the skin. In addition, the retention of stratum corneum cells that peel off from the surface of the stratum corneum hinders the smooth excretion of melanin in the epidermis, causing pigmentation and dullness of the skin. Furthermore, it is also known that wound healing of the epidermis is delayed. In order to prevent or improve the progression of these phenomena, many attempts have been made to search for components that promote proliferation of epidermal keratinocytes and to propose topical preparations for the skin.

Barley (scientific name: Hordeum vulgare L.) belongs to the genus Barley, belonging to the family Gramineae, and is one of the crops cultivated from the earliest times in the world. Until now, barley has been reported that the young leaf extract has an antioxidant effect (Patent Document 7), and the extract of purple barley or black barley, which is a colored line of barley, has a melanin production inhibitory effect and a ceramide production promotion effect (Patent Document 7). Reference 8) is known. However, it has not been known that an extract of germinated purple barley seeds has collagen production-promoting action, MMP-inhibiting action, hyaluronic acid production-promoting action and cell proliferation-promoting action.

JP-A-2000-119125 JP 2006-199611 A JP-A-3-44331 JP-A-2003-137801 JP-A-5-186368 JP 2007-1924 A JP-A-6-122619 JP 2008-143827 A

"Matrix Metalloprotease in Gastrointestinal Cancer", Japanese Society of Gastroenterology, 100, 152, 2003 "Arthritis Rheumatism", Vol. 10, pp 357, 1967 "Combination Composition", Kanehara Shuppan, Item 481, 1984 "Inflammation", Japanese Inflammation Society, Vol. 11, Item 16, 1991 Varani J et al. , J Invest Dermatol, Vol. 3, pp 57-60, 1998

There is a demand for materials that are safe, have excellent stability, and have excellent collagen production-promoting action, MMP-inhibiting action, hyaluronic acid production-promoting action, and cell proliferation-promoting action, but none have yet been provided that are sufficiently satisfactory. This is the current situation.

Under these circumstances, the present inventors have made extensive studies and found that the extract of germinated purple barley seeds has excellent collagen production-promoting action, MMP inhibitory action, hyaluronic acid production-promoting action and cell proliferation-promoting action. It was found that the stability was also excellent. Furthermore, the external preparation or internal preparation containing the extract is safe and stable, and has excellent collagen production-promoting action, MMP inhibitory action, hyaluronic acid production-promoting action, and cell proliferation-promoting action.・We found that it can be used as a health material and medicine, and completed the present invention.

That is, the present invention includes the following inventions.
(1) A collagen production promoter characterized by containing an extract of germinated purple barley seeds.
(2) An MMP inhibitor comprising an extract of germinated purple barley seeds.
(3) A hyaluronic acid production promoter characterized by containing an extract of germinated purple barley seeds.
(4) A cell growth promoter characterized by containing an extract of germinated purple barley seeds.
(5) A wrinkle reducing agent characterized by containing an extract of germinated purple barley seeds.
(6) A food composition for preventing or improving various diseases caused by MMP enhancement, characterized by containing an extract of germinated purple barley seeds.

According to the present invention, there are provided collagen production promoters, MMP inhibitors, hyaluronic acid production promoters and cell proliferation promoters containing extracts of germinated purple barley seeds as active ingredients.

The purple barley used in the present invention refers to barley (scientific name: Hordeum vulgare L.) with purple glumes, glumes, stems or leaves, and is a grain belonging to the genus Barley of the family Gramineae. Examples of barley varieties include OUC321, CI158, and CI244.

The purple barley seeds used in the present invention can be obtained by cultivating them, can be purchased commercially, or can be obtained from the growing area.

In the present invention, germinated purple barley seeds are used as raw materials for extraction. Here, "germination" is a term that includes rooting, and "germination state" refers to a state in which a germination and/or radicle has emerged from a seed. In the present invention, "germinated purple barley seeds" include germinated seeds of purple barley, malt (seeds obtained by cutting and drying germinated barley roots and buds), germinated seedlings (sprouts), and the like. subsumed. Here, germinated seeds generally refer to those having roots, hypocotyls, cotyledons and endosperms, and seedlings (sprouts) generally refer to seeds that have germinated and grown to the point where they have no endosperm. Say.

As for the appearance of the germinated purple wheat seeds used as the raw material for extraction in the present invention, it is preferable that the purple wheat seeds have extended roots and sprouts, and the whole seeds including the roots and sprouts can be used. As for the size of germinated purple wheat seeds, the sprout length is preferably 0.1 to 80 mm, more preferably 1 to 50 mm. The root length is preferably 1 to 200 mm, more preferably 10 to 100 mm.

The germinated purple wheat seeds can be prepared by drying the seeds after germination. For example, the germination treatment can be carried out by immersing raw or dried purple barley seeds in water to break dormancy, then sowing the seeds in a seed bed and cultivating the seeds while watering them so that the seeds do not dry out completely. . The immersion temperature is preferably 2 to 40°C, more preferably 10 to 30°C. The soaking time of seeds is preferably 0.5 to 24 hours, more preferably 2 to 12 hours. The seeding amount can be exemplified from 0.1 to 5 grains/cm ² per cultivation area. Non-woven fabrics other than soil, towel paper, sponge, vermiculite, perlite, and the like can be used as the seed bed. Moreover, it is more preferable to cover with towel paper or the like soaked with water for moisturizing, and further cover with food wrap film or the like. The cultivation temperature is preferably 10 to 30°C, more preferably 20 to 25°C. The cultivation period varies depending on temperature and light irradiation conditions, but is about 2 to 10 days.

For the drying treatment, a method commonly used for drying plants can be used. Examples include natural drying (air drying), sun drying, dry heat drying, air drying, hot air drying, spray drying, reduced pressure drying, vacuum drying, etc. Dry heat drying, air drying and hot air drying are preferred. The temperature and time conditions of the drying treatment are not particularly limited, but the temperature is preferably 40 to 70 ° C., and the time varies depending on the drying temperature, the moisture content of the germinated purple wheat seeds, and the total amount to be dried, but is generally 4 to 24 hours. is in the range of

In the present invention, for the extraction of germinated purple wheat seeds (hereinafter referred to as "germinated purple wheat seeds"), the seeds or the plant body containing the seeds may be used as they are, and dried, pulverized, and chopped. etc. may be performed.

The extraction method using a solvent is not particularly limited, and may be carried out, for example, by heat extraction, normal temperature extraction, low temperature extraction, stirring extraction, column extraction, or the like. Examples of extraction solvents include water, lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, 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.), hydrocarbons (hexane, heptane, liquid paraffin, etc.), ethers (ethyl ether, tetrahydrofuran, propyl ether etc.). Polar solvents such as water, lower alcohols and liquid polyhydric alcohols are preferred, and water, ethanol, 1,3-butylene glycol and propylene glycol are particularly preferred. These solvents may be used singly or in combination of two or more. Particularly preferred extraction solvents include water, a mixed polar solvent of water-ethanol, or a mixed polar solvent of water-1,3-butylene glycol. Among them, it preferably contains 20 to 100% by weight of ethanol or 1,3-butylene glycol, and most preferably 50 to 100% by weight. Also, a solvent whose pH is adjusted by adding an acid or an alkali to the extraction solvent can be used.

The amount of the solvent to be used is not particularly limited. For example, it may be 5 times or more, preferably 10 times or more, relative to the germinated purple wheat seeds (dry weight), but concentration or isolation may be performed after extraction. It is preferably 100 times or less for convenience of operation in the case. Also, the extraction temperature and time can be appropriately selected depending on the type of solvent used, the pressure during extraction, and the like.

The above extract may be used as an extracted solution, but if necessary, concentration (concentration by vacuum concentration, membrane concentration, etc.), dilution, filtration, decolorization by activated carbon, etc., within the range where the effect of the present invention is exhibited. , deodorization, ethanol precipitation, or the like may be performed before use. Furthermore, the extracted solution may be subjected to a treatment such as concentration to dryness, spray drying, freeze drying, etc., and used as a dried product.

In the present invention, the above extract may be used as it is, and oils and fats, waxes, hydrocarbons, etc., which are ingredients used in cosmetics, quasi-drugs, pharmaceuticals, foods, etc., as long as the effects of the extract are not impaired. fatty acids, alcohols, esters, surfactants, metallic soaps, pH adjusters, preservatives, fragrances, moisturizers, powders, UV absorbers, thickeners, pigments, antioxidants, whitening agents, Components such as chelating agents, excipients, film-forming agents, sweeteners and acidulants may be contained.

The present invention can be used for any of cosmetics, quasi-drugs, pharmaceuticals, and foods. , bath agents, foundations, powders, lipsticks, ointments, poultices, tablets, chocolates, gums, candies, beverages, powders, granules, tablets, sugar-coated tablets, capsules, syrups, pills, suspensions, liquids , emulsions, suppositories, injection solutions and the like.

For external use, the content of the extract used in the present invention is preferably 0.0001% by weight or more, more preferably 0.001 to 10% by weight in terms of solid matter. Furthermore, 0.01 to 5% by weight is most preferred. If it is less than 0.0001% by weight, it is difficult to expect a sufficient effect. If it exceeds 10% by weight, it is difficult to recognize the enhancement of the effect and it is uneconomical.

In the case of internal use, the intake varies depending on age, body weight, symptoms, therapeutic effect, administration method, treatment time, and the like. Generally, the daily intake per adult is preferably 5 mg or more, more preferably 10 mg to 5 g. Furthermore, 20 mg to 2 g is most preferred.

Next, in order to explain the present invention in detail, production examples, experimental examples and formulation examples of the extract used in the present invention will be given as examples, but the present invention is not limited to these. Unless otherwise specified, % shown in Examples means % by weight.

(1) Preparation example of germinated purple barley seeds Germinated purple barley seeds used as raw materials for extraction were prepared as follows. 100 g of dried purple barley seeds were soaked in 500 mL of tap water at 25° C. for 6 hours, the soaked water was discarded, and the purple barley seeds were wrapped in a paper towel moistened with water and covered with a food wrap film. After germination by moisturizing at room temperature for 4 days, dry heat drying was performed at 60° C. for 5 hours to obtain 97 g of germinated purple barley seeds. At this time, the sprout length was 8 mm, and the root length was 23 mm.

(2) Production Example of Extract In the production of the following extract, when the raw material for extraction was germinated purple barley seeds, the germinated purple barley seeds prepared in (1) were used.

(Production Example 1) Production of hot water extract of germinated purple barley seeds 200 mL of purified water was added to 10 g of dried germinated purple barley seeds, and extraction was carried out at 95 to 100°C for 2 hours. The resulting extract was filtered and then freeze-dried to obtain 0.98 g of a hot water extract of germinated purple barley seeds.

(Production Example 2) Production of 50% Ethanol Extract of Germinated Purple Barley Seeds To 10 g of dried germinated purple barley seeds, 200 mL of a 50% ethanol aqueous solution was added, and the mixture was immersed at room temperature for 7 days for extraction. After the resulting extract was filtered, the filtrate was concentrated and freeze-dried to obtain 1.00 g of a 50% ethanol extract of germinated purple barley seeds.

(Production Example 3) Production of Ethanol Extract of Germinated Purple Barley Seeds 200 mL of ethanol was added to 10 g of dried germinated purple barley seeds, and the mixture was immersed at room temperature for 7 days for extraction. After the resulting extract was filtered, the filtrate was concentrated and freeze-dried to obtain 0.24 g of an ethanol extract of germinated purple barley seeds.

(Production Example 4) Preparation of 1,3-butylene glycol extract of germinated purple barley seeds 200 mL of 1,3-butylene glycol was added to 10 g of dried germinated purple barley seeds, and the mixture was immersed at room temperature for 7 days for extraction. rice field. The resulting extract was filtered to obtain 187 g of a 1,3-butylene glycol extract of germinated purple barley seeds.

(Comparative Production Example 1) Production of hot water extract of ungerminated purple wheat seeds To 10 g of dried purple wheat seeds, 200 mL of purified water was added, and extraction was performed at 95 to 100°C for 2 hours. The resulting extract was filtered and then freeze-dried to obtain 0.90 g of a hot water extract of ungerminated purple barley seeds.

(Comparative Production Example 2) Production of 50% ethanol extract of ungerminated purple wheat seeds To 10 g of dried purple wheat seeds, 200 mL of a 50% ethanol aqueous solution was added, and the mixture was immersed at room temperature for 7 days for extraction. After the resulting extract was filtered, the filtrate was concentrated and freeze-dried to obtain 1.12 g of a 50% ethanol extract of ungerminated purple barley seeds.

(Comparative Production Example 3) Production of ethanol extract of ungerminated purple wheat seeds 200 mL of ethanol was added to 10 g of dried purple wheat seeds, and the mixture was immersed at room temperature for 7 days for extraction. After the resulting extract was filtered, the filtrate was concentrated and freeze-dried to obtain 0.33 g of an ethanol extract of ungerminated purple barley seeds.

(Prescription example 1) Lotion Prescription Content (%)
1. Hot water extract of germinated purple barley seeds (Production Example 1) 2.0
2.1,3-butylene glycol 8.0
3. Glycerin 2.0
4. Xanthan gum 0.02
5. Citric acid 0.01
6. Sodium citrate 0.1
7. Ethanol 5.0
8. Methyl paraoxybenzoate 0.1
9. Polyoxyethylene hydrogenated castor oil (40E.O.) 0.1
10. Fragrance Appropriate amount 11. Adjust the total amount to 100 with purified water [Manufacturing method] Components 1 to 6 and 11 and components 7 to 10 are uniformly dissolved, mixed and filtered to obtain a product.

(Comparative Formulation Example 1) Conventional Lotion A conventional lotion was prepared by replacing the hot water extract of germinated purple barley seeds (Production Example 1) with purified water in Formulation Example 1.

(Prescription example 2) Cream Prescription Content (%)
1. 50% ethanol extract of germinated purple barley seeds (Production Example 2) 1.0
2. Squalane 5.5
3. Olive oil 3.0
4. stearic acid 2.0
5. Beeswax 2.0
6. Octyldodecyl myristate 3.5
7. Polyoxyethylene cetyl ether (20 E.O.) 3.0
8. behenyl alcohol 1.5
9. Glyceryl monostearate 2.5
10. Perfume 0.1
11. Methyl paraoxybenzoate 0.2
12.1,3-butylene glycol 8.5
13. The total amount is adjusted to 100 with purified water [Manufacturing method] Components 2 to 9 are dissolved by heating, mixed, and maintained at 70°C to form an oil phase. Ingredients 1 and 11 to 13 are dissolved by heating and mixed, and kept at 75° C. to form an aqueous phase. Add the water phase to the oil phase to emulsify, cool while stirring, add component 10 at 45°C, and further cool to 30°C to obtain the product.

(Comparative Formulation Example 2) Conventional Cream In Formulation Example 2, a conventional cream was obtained by replacing the 50% ethanol extract of germinated purple barley seeds (Production Example 2) with purified water.

(Prescription example 3) Milky lotion Prescription Content (%)
1. Ethanol extract of germinated purple barley seeds (Production Example 3) 0.01
2. Squalane 5.0
3. Olive oil 5.0
4. Jojoba oil 5.0
5. Cetanol 1.5
6. Glyceryl monostearate 2.0
7. Polyoxyethylene cetyl ether (20 E.O.) 3.0
8. Polyoxyethylene sorbitan monooleate (20E.O.) 2.0
9. Perfume 0.1
10. Propylene glycol 1.0
11. Glycerin 2.0
12. Methyl paraoxybenzoate 0.2
13. Adjust the total amount to 100 with purified water [Manufacturing method] Components 1 to 8 are dissolved by heating, mixed, and kept at 70°C to form an oil phase. Ingredients 10 to 13 are dissolved by heating, mixed, and kept at 75° C. to form an aqueous phase. Add the water phase to the oil phase to emulsify, cool while stirring, add component 9 at 45°C, and further cool to 30°C to obtain the product.

(Prescription example 4) Gel Formulation Content (%)
1. 1,3-butylene glycol extract of germinated purple barley seeds (Production Example 4) 1.0
2. Ethanol 5.0
3. Methyl paraoxybenzoate 0.1
4. Polyoxyethylene hydrogenated castor oil (60E.O.) 0.1
5. Perfume appropriate amount 6.1,3-butylene glycol 5.0
7. Glycerin 5.0
8. Xanthan gum 0.1
9. Carboxy vinyl polymer 0.2
10. Potassium hydroxide 0.2
11. The total amount is adjusted to 100 with purified water [Manufacturing method] Components 2 to 5 and components 1 and 6 to 11 are uniformly dissolved and mixed to obtain a product.

(Prescription example 5) Pack Prescription Content (%)
1. Hot water extract of germinated purple barley seeds (Production Example 1) 1.0
2. 1,3-butylene glycol extract of germinated purple barley seeds (Production Example 4) 5.0
3. Polyvinyl alcohol 12.0
4. Ethanol 5.0
5.1,3-butylene glycol 8.0
6. Methyl paraoxybenzoate 0.2
7. Polyoxyethylene hydrogenated castor oil (20E.O.) 0.5
8. Citric acid 0.1
9. Sodium citrate 0.3
10. Fragrance Appropriate amount 11. Adjust the total amount to 100 with purified water [Manufacturing method] Components 1 to 11 are uniformly dissolved to obtain a product.

(Prescription example 6) Foundation Prescription Content (%)
1. 50% ethanol extract of germinated purple barley seeds (Production Example 2) 1.0
2. stearic acid 2.4
3. Polyoxyethylene sorbitan monostearate (20E.O.) 1.0
4. Polyoxyethylene cetyl ether (20 E.O.) 2.0
5. Cetanol 1.0
6. Liquid Lanolin 2.0
7. Liquid paraffin 3.0
8. Isopropyl myristate 6.5
9. Carboxymethylcellulose sodium 0.1
10. Bentonite 0.5
11. Propylene glycol 4.0
12. Triethanolamine 1.1
13. Methyl paraoxybenzoate 0.2
14. Titanium dioxide 8.0
15. Talc 4.0
16. Bengara 1.0
17. Yellow iron oxide 2.0
18. Perfume Appropriate amount 19. The total amount is adjusted to 100 with purified water [Manufacturing method] Components 2 to 8 are dissolved by heating and maintained at 80°C to form an oil phase. Ingredient 19 is sufficiently swelled with ingredient 9, then ingredients 1 and 10 to 13 are added and mixed uniformly. Ingredients 14 to 17 pulverized and mixed with a pulverizer are added to this, stirred with a homomixer and kept at 75° C. to form an aqueous phase. Add the water phase to the oil phase with stirring to emulsify. After cooling, add component 18 at 45°C and cool to 30°C with stirring to obtain the product.

(Prescription example 7) Bath agent Prescription Content (%)
1. Ethanol extract of germinated purple barley seeds (Production Example 3) 1.0
2. Sodium bicarbonate 50.0
3. Yellow No. 202 (1) Appropriate amount 4. Perfume Appropriate amount 5. [Manufacturing method] Components 1 to 5 are uniformly mixed to obtain a product.

(Prescription example 8) Ointment Prescription Content (%)
1. Hot water extract of germinated purple barley seeds (Production Example 1) 5.0
2. 1,3-butylene glycol extract of germinated purple barley seeds (Production Example 4) 1.0
3. Polyoxyethylene cetyl ether (30 E.O.) 2.0
4. Glyceryl monostearate 10.0
5. Liquid paraffin 5.0
6. Cetanol 6.0
7. Methyl paraoxybenzoate 0.1
8. Propylene glycol 10.0
9. The total amount is adjusted to 100 with purified water [Manufacturing method] Components 3 to 6 are dissolved by heating, mixed, and kept at 70°C to form an oil phase. Ingredients 1, 2 and 7 to 9 are dissolved by heating, mixed, and maintained at 75°C to form an aqueous phase. The water phase is added to the oil phase to emulsify, and the mixture is cooled to 30°C while stirring to obtain a product.

(Prescription Example 9) Powder Formulation Content (%)
1. Hot water extract of germinated purple barley seeds (Production Example 1) 1.0
2. Dry cornstarch 39.0
3. Microcrystalline cellulose 60.0
[Manufacturing method] Components 1 to 3 are mixed to form a powder.

(Prescription Example 10) Tablet Prescription Content (%)
1. Ethanol extract of germinated purple barley seeds (Production Example 3) 5.0
2. Dry cornstarch 25.0
3. Carboxymethylcellulose calcium 20.0
4. Microcrystalline cellulose 40.0
5. polyvinylpyrrolidone 7.0
6. Talc 3.0
[Manufacturing method] Components 1 to 4 are mixed, and then an aqueous solution of component 5 is added as a binder to form granules. Ingredient 6 is added to the molded granules and pressed into tablets. One tablet is 0.52 g.

(Prescription example 11) Tablet prescription Content (%)
1. Ethanol extract of germinated purple barley seeds (Production Example 3) 2.0
2. Dry cornstarch 49.8
3. Erythritol 40.0
4. Citric acid 5.0
5. Sucrose fatty acid ester 3.0
6. Perfume 0.1
7. Purified water 0.1
[Manufacturing method] Components 1 to 4 and 7 are mixed and granulated. Ingredients 5 and 6 are added to the molded granules and compressed into tablets. 1 grain is 1.0 g.

(Prescription Example 12) Beverage Prescription Content (%)
1. Hot water extract of germinated purple barley seeds (Production Example 1) 0.05
2. Stevia 0.05
3. Malic acid 5.0
4. Perfume 0.1
5. Adjust the total amount to 100 with purified water [Manufacturing method] Components 1 to 3 are dissolved in a small amount of water. Ingredients 4 and 5 are then added and mixed.

Next, experimental examples will be given in order to describe the effects of the present invention in detail.

Experimental Example 1 Measurement of Type I Collagen (COL1A1), MMP-1, MMP-2 and Hyaluronic Acid Synthase 2 (HAS2) mRNA Expression Levels COL1A1, MMP-1, MMP-2 and HAS2 mRNA expression levels were measured. . 1×10 ⁵ human skin fibroblasts were seeded in a φ60 mm dish and cultured in a DMEM culture solution containing 10% FBS under conditions of 37° C. and 5% CO ₂ . When the cells became confluent, each sample was cultured for 24 hours in DMEM(-) culture medium added to final concentrations of 1, 10, and 100 μg/mL, and then total RNA was extracted. Total RNA was extracted from the cells using RNAiso Plus (Takara Bio), and the amount of total RNA was determined by absorbance at 260 nm using a spectrophotometer (Nanodrop). The mRNA expression level was measured by real-time RT-PCR method based on the total RNA extracted from the cells. For the real-time RT-PCR method, High Capacity RNA-to-cDNA Kit (Applied Biosystems) and SYBR Select Master Mix (Applied Biosystems) were used. That is, after reverse transcription of 500 ng of total RNA, PCR reaction (95°C: 15 seconds, 60°C: 60 seconds, 40 cycles) was performed. Other operations were carried out according to a prescribed method, and the expression levels of COL1A1, MMP-1, MMP-2 and HAS2 mRNA were determined as a ratio to the expression level of β-actin mRNA as an internal standard. The COL1A1 expression promotion rate was calculated as the ratio of the COL1A1 mRNA expression level in the sample-added group to the COL1A1 mRNA expression level in the control (sample-free) group. The MMP-1 expression suppression rate, MMP-2 expression suppression rate, and HAS2 expression promotion rate were also calculated in the same manner. For MMP-1 and MMP-2, the concentration required to suppress the mRNA expression level to 80% (hereinafter referred to as IC ₈₀ ) was calculated from the test results. Lower IC ₈₀ values correspond to more potent MMP-1 and MMP-2 inhibitors. For some materials, increasing the concentration to solubility does not lead to an _IC80 calculation. The primers used for measuring the expression level of each gene are as follows.

Primer set AGGACAAGAGGGCATGTCTGGTT for COL1A1 (SEQ ID NO: 1)
TTGCAGTGGGTAGGTGATGTTCTG (SEQ ID NO: 2)
Primer set GGGAGATCATCGGGACAACTC for MMP-1 (SEQ ID NO: 3)
TGAGCATCCCCTCCAATACC (SEQ ID NO: 4)
Primer set CCGTCGCCCATCATCAA for MMP-2 (SEQ ID NO: 5)
CTTCTGCATCTTCTTTAGTGTGTCCTT (SEQ ID NO: 6)
Primer set TGGATGACCTACGAAGCGATTA for HAS2 (SEQ ID NO: 7)
GCTGGATTACTGTGGCAATGAG (SEQ ID NO: 8)
Primer set CACTCTTCCAGCCTTCCTTCC for β-actin (SEQ ID NO: 9)
GTGTTGGGCGTACAGGTCTTTG (SEQ ID NO: 10)

The results of these experiments are shown in Tables 1-4. As a result, the extract of germinated purple barley seeds of the present invention has excellent COL1A1 expression promoting effect (collagen production promoting effect), MMP-1 expression suppressing effect (MMP-1 inhibitory effect), MMP-2 expression suppressing effect ( MMP-2 inhibitory effect) and HAS2 expression promoting effect (hyaluronic acid production promoting effect) were observed. In particular, the HAS2 expression-promoting effect of the hot water extract of germinated purple barley seeds (Production Example 1), the COL1A1 expression-promoting effect and the HAS2 expression-promoting effect of the 50% ethanol extract of germinated purple barley seeds (Production Example 2), and the germinated purple barley seeds. In the MMP-1 expression-suppressing effect, MMP-2 expression-suppressing effect, and HAS2 expression-promoting effect of the ethanol extract of seeds (Production Example 3), the conventional hot water extract of ungerminated purple barley seeds (Comparative Production Example 1), Compared to the 50% ethanol extract of ungerminated purple barley seeds (Comparative Production Example 2) and the ethanol extract of ungerminated purple barley seeds (Comparative Production Example 3), the effect was significantly higher.

Experimental Example 2 Cell Proliferation Promotion Test Human-derived keratinocytes were seeded in a DMEM culture medium containing 0.2% FBS at 1×10 ³ cells per well in a 96-well plate, and the final concentration of each sample was 10 μg/mL. and cultured for 5 days under conditions of 37°C and 5% _CO2 . Cell count was measured by staining method. That is, after the culture was completed, the culture medium was removed, and the cells were fixed using methanol. Subsequently, 0.1% methylene blue was added to stain the cells for 1 hour. After drying, 100 μL of 0.1N HCl was added to each well and well stirred, and the absorbance at 650 nm was measured using a microplate reader. The cell growth rate was calculated as the ratio of the number of cells in the sample-added group to the number of cells in the control (no sample added) group.

These experimental results are shown in Table 5. As a result, the extract of germinated purple barley seeds of the present invention exhibited an excellent cell growth-promoting action. In particular, the cell growth-promoting effect of the 50% ethanol extract of germinated purple wheat seeds (Production Example 2) and the ethanol extract of germinated purple wheat seeds (Production Example 3) is higher than that of the conventional 50% ethanol extraction of ungerminated purple wheat seeds. (Comparative Production Example 2) and ethanol extract of ungerminated purple barley seeds (Comparative Production Example 3).

Experimental Example 3 Use Test Using the cream of Formulation Example 2 and the conventional cream of Comparative Formulation Example 2, a one-month use test was conducted on 5 persons (25 to 65 years old) with wrinkles and sagging. After use, the degree of wrinkles and sagging was determined by a questionnaire.

As a result, the cream containing the extract of the present invention reduced wrinkles and sagging. During the test period, no skin trouble occurred, and there was no problem in terms of safety. Moreover, there was no problem with deterioration of prescription components.

Further, using the lotion of Formulation Example 1 and the conventional lotion of Comparative Formulation Example 1, a use test was conducted in the same manner. As a result, wrinkles and sagging were reduced by the lotion containing the extract of the present invention.

As described above, the extract of germinated purple barley seeds of the present invention has excellent collagen production-promoting action, MMP-inhibiting action, hyaluronic acid production-promoting action and cell proliferation-promoting action, and is also excellent in stability. Therefore, the extract of germinated purple barley seeds of the present invention can be used not only in the cosmetic field such as skin aging, but also in the medical field such as suppression of functional deterioration due to aging, prevention and treatment of cancer, etc. It is expected to be applied to quasi-products and pharmaceuticals.

Claims (6)

A collagen production promoter comprising an extract of germinated purple barley seeds. An MMP inhibitor comprising an extract of germinated purple wheat seeds. A hyaluronic acid production promoter comprising an extract of germinated purple barley seeds. A cell growth promoting agent comprising an extract of germinated purple wheat seeds. A wrinkle-improving agent comprising an extract of germinated purple barley seeds. A food composition for preventing or improving various diseases caused by MMP enhancement, characterized by containing an extract of germinated purple barley seeds.