JP7597350B2 - Skin preparations for external and internal use - Google Patents

Description

The present invention relates to MMP inhibitors, collagen production promoters, wrinkle improvement agents, hyaluronic acid production promoters, antioxidants, skin whitening agents, pharmaceuticals, and food compositions.

Skin is exposed to various physical and chemical stresses such as ultraviolet rays, dryness, cold, heat, and drugs every day. As a result, the skin's functions are impaired, 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 lines, and are temporary wrinkles that occur when the moisture content in the epidermal stratum corneum decreases due to dry skin. On the other hand, dermal wrinkles are wrinkles that are formed by ultraviolet rays contained in sunlight and aging. The mechanism of their formation includes a decrease in the collagen synthesis ability of dermal fibroblasts due to ultraviolet rays and aging, and the promotion of collagen decomposition due to an increase in matrix proteinase (MMP).

Epidermal wrinkles caused by dryness and dermal wrinkles have different histological morphology, onset mechanisms, and treatment methods, and dermal wrinkles caused by ultraviolet rays and aging are difficult to improve with the use of cosmetics with moisturizing effects.

To date, there have been reports of an agent for preventing and improving skin wrinkles, which contains hydrolyzed almonds as an active ingredient, (Patent Document 1), and an agent for improving wrinkles caused by ultraviolet radiation, which contains extracts of Jochou-kei, Tensho and Xanthomonas japonica as active ingredients (Patent Document 2).

The dermis also contains fibroblasts and collagen, with type I collagen accounting for 80% of the total. In addition to type I collagen, types III, V, XII, and XIV are known to exist. One of the causes of wrinkles and sagging skin is a decrease in type I collagen. Therefore, promoting the production of type I collagen is thought to be effective in preventing and improving wrinkles and sagging skin. Promoting the production of type I collagen is also effective in improving wound healing in the skin.

Collagen is the main structural protein that accounts for approximately one-third of mammalian tissues, and is an essential component of many matrix tissues, such as cartilage, bone, tendon, and skin. When collagenase (MMP-1), which belongs to the MMPs, cleaves collagen molecules, which are stable in normal tissues, denaturing them into single-stranded gelatin that can be degraded by various other proteases. As a result, the structural integrity of the matrix tissue is lost.

Gelatinase (MMP-2), which belongs to MMP, is an enzyme produced by fibroblasts, endothelial cells, cancer cells, etc., and breaks down substrates such as collagen, gelatin, and elastin (structural proteins that are special components of elastic tissues such as arteries, bones, and skin). Therefore, substances that have inhibitory activity against gelatinase are expected to have the effect of suppressing angiogenesis and cancer metastasis in cancer tissue, and are considered to be useful in the prevention and treatment of cancer diseases. Furthermore, inhibition of MMP is useful not only for cancer diseases, but also for the prevention, treatment, and improvement of ulcer formation, chronic rheumatoid arthritis, osteoporosis, periodontal inflammation, and various diseases caused by increased MMP.

Materials that have been proposed as having collagenase inhibitory activity include, for example, cacao husk extract (Patent Document 3), which is the skin of the cacao bean, Rubus fragilis extract (Patent Document 4), and lactoferrin (Patent Document 5). In a situation where interest in skin aging and oral hygiene is increasing, there is a demand to find materials that have no side effects, are highly safe, and have excellent collagenase activity inhibitory activity.

Fibroblasts also produce proteins such as collagen and glycosaminoglycans such as hyaluronic acid to form dermal connective tissue, which maintains skin firmness. It is believed that when this connective tissue loses its contractile force and further its elasticity, wrinkles and sagging of the skin occur.

In particular, hyaluronic acid is known as a polymeric polysaccharide widely distributed in connective tissues, and takes on a gel-like form in the dermis, maintaining the elasticity of the skin. Therefore, the deterioration and reduction of hyaluronic acid are considered to be important in skin aging. In addition, since hyaluronic acid is a polymer, there is a problem that cosmetics containing it are not easily absorbed even when directly applied to the skin. Therefore, up to now, a skin topical agent that can promote the production of collagen and hyaluronic acid by activating fibroblasts has been sought (Patent Document 6). In addition, hyaluronic acid is also present in joints, and is known to function to cushion the impact of load on the joints and to smooth the movement of the joints. In the case of joint diseases such as osteoarthritis, chronic rheumatoid arthritis, suppurative arthritis, gouty arthritis, traumatic arthritis, and osteoarthritis, it is known that the amount of hyaluronic acid in the synovial fluid decreases or decreases with age. In such joint diseases, it is believed that increasing the amount of hyaluronic acid in the synovial fluid is effective in improving lubrication, covering and protecting the articular cartilage, suppressing pain, and improving or normalizing pathological synovial fluid. For example, it is known that when sodium hyaluronate is injected into the joints of patients with chronic rheumatoid arthritis, traumatic arthritis, osteoarthritis, and osteoarthritis, the above-mentioned symptoms are improved. However, these treatments take a long time. Therefore, there is a demand for foods and medicines that contain hyaluronic acid production promoters so that prevention and treatment can be easily carried out in daily life.

Floaters are a symptom where a thin shadow resembling a lint or a mosquito appears in the field of vision. They occur when opacity in the vitreous body, which fills the inside of the eye, casts a shadow on the retina. Floaters can be broadly divided into two types: physiological floaters, which develop due to aging, ultraviolet rays, active oxygen, etc., and pathological floaters, which appear as a symptom of diseases such as retinal detachment, retinal tear, vitreous hemorrhage, and uveitis. Physiological floaters are caused by liquefaction due to a decrease in hyaluronic acid, the main component of the vitreous body, and the resulting breakdown of collagen fibers, causing opacity in the vitreous body. Treatments include vitrectomy surgery and laser treatment, but these procedures are not often performed in Japan due to safety concerns, and treatment abroad requires a large amount of money. Therefore, in order to prevent and improve physiological floaters, foods and medicines containing hyaluronic acid production promoters that can be used on a daily basis are desired.

Generally, skin pigmentation such as age spots, freckles, and sunburn is thought to be caused by hormonal abnormalities and UV stimulation causing melanin pigment to be produced excessively by melanin-producing cells in the skin, which then deposits in the skin. One method for preventing such pigmentation is known to be the suppression of excessive melanin production. Conventionally, ascorbic acid (vitamin C) and the like have been used internally and externally to treat pigmentation (Patent Document 7).

The skin is located in the outermost layer of the body, and is an organ that is prone to the generation of active oxygen due to the effects of ultraviolet rays, etc., and is constantly exposed to oxygen stress. On the other hand, active oxygen scavenging enzymes exist in skin cells, and they protect skin cells from damage caused by active oxygen unless the generation of active oxygen exceeds their capacity. However, it is known that the activity of active oxygen scavenging enzymes in skin cells decreases with age, and when damage caused by active oxygen overcomes the defense reaction, the skin is oxidized, the cell function deteriorates, and aging progresses. In addition, it is thought that when organs other than the skin are exposed to active oxygen that exceeds their active oxygen scavenging capacity, their function decreases, leading to aging, and various lifestyle-related diseases such as cancer and myocardial infarction develop. Therefore, active oxygen scavengers and antioxidants have been studied for the purpose of protecting against damage caused by active oxygen, and foods, cosmetics, quasi-drugs, and medicines containing active oxygen scavenging enzymes such as SOD and catalase, active oxygen scavengers such as SOD-like active substances, and antioxidants have been developed (Patent Documents 8 and 9).

It is known that the vegetable oil and fermented liquid of the fruits of the Rowan tree (Sorbus aucuparia), which belongs to the family Rosaceae in the order Rosales, are used as hair and nail conditioning agents (Patent Documents 10 and 11). However, it was not known that the fruits of the Rowan tree have MMP inhibitory effects, collagen production promotion effects, hyaluronic acid production promotion effects, antioxidant effects, and melanin production suppression effects.

JP 2000-119125 A JP 2006-199611 A Japanese Patent Application Publication No. 3-44331 JP 2003-137801 A Japanese Patent Application Publication No. 5-186368 JP 2007-1924 A Japanese Patent Application Publication No. 05-229931 Japanese Patent Application Publication No. 9-118630 Japanese Patent Application Publication No. 9-208484 Special table 2019-510803 publication Special Publication No. 2015-503587

There is a demand for a material that is safe and stable, and has excellent MMP inhibitory effects, collagen production promotion effects, hyaluronic acid production promotion effects, antioxidant effects, and melanin production suppression effects, but currently no fully satisfactory material has been provided.

The inventors conducted intensive research to solve this problem, and as a result, discovered that Rowan fruit extract has excellent MMP inhibitory activity, collagen production promotion activity, hyaluronic acid production promotion activity, antioxidant activity, and melanin production inhibitory activity, and is also excellent in stability. Furthermore, they discovered that topical or internal preparations containing the extract are safe and stable, and have excellent MMP inhibitory activity, collagen production promotion activity, hyaluronic acid production promotion activity, antioxidant activity, and melanin production inhibitory activity, and can be used as multifunctional beauty and health materials and medicines, leading to the completion of the present invention.

That is, the present invention comprises the following (1) to (8).

(1) An MMP inhibitor characterized by containing an extract of Rowan fruit.
(2) A collagen production promoter characterized by containing an extract of Rowan fruit.
(3) A wrinkle improving agent characterized by containing an extract of Rowan fruit.
(4) A hyaluronic acid production promoter characterized by containing an extract of Rowan fruit.
(5) An antioxidant characterized by containing an extract of Rowan fruit.
(6) A skin whitening agent characterized by containing an extract of Rowan fruit.
(7) A pharmaceutical comprising an extract of Rowan fruit.
(8) A food composition for preventing and improving various diseases, joint diseases and physiological floaters caused by increased MMP activity, characterized by containing an extract of Rowan fruit.

The present invention is described in detail below.

The European rowan (scientific name: Sorbus aucuparia) used in this invention is a deciduous tree native to Europe that belongs to the genus Sorbus and the family Rosaceae, and is a medium-sized shrub that grows to about 8 to 10 meters. The fruit is a pome-shaped fruit that changes from green to bright red as it ripens in the summer, and is primarily used for food.

The Rowan fruit used in the present invention can be obtained from the growing area or purchased commercially. The fruit itself can be extracted as is, or it can be processed by drying, crushing, shredding, etc.

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.), and 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 alone or in combination. Particularly preferred extraction solvents include water, ethanol, 1,3-butylene glycol, mixed polar solvents of water-ethanol, or mixed polar solvents of water-1,3-butylene glycol. There are no particular limitations on the amount of solvent used, and it is sufficient to use an amount of solvent that is at least 10 times, preferably at least 20 times, the amount of Rowan fruit (dry weight), but it is preferable to use an amount of solvent that is no more than 100 times for convenience of operations when concentrating or isolating after extraction. In addition, the extraction temperature and time can be appropriately selected depending on the type of solvent used, the pressure during extraction, etc.

The extraction method is not particularly limited, and may be performed by methods such as heating extraction, room temperature extraction, low temperature extraction, stirring extraction, or column extraction. The extract may also be used in combination with the fruit.

The above extract may be used as it is in the form of an extracted solution, but if necessary, it may be used after being subjected to processes such as concentration (vacuum concentration, membrane concentration, etc.), dilution, filtration, decolorization with activated carbon, deodorization, ethanol precipitation, etc., within the scope of the effects of the present invention. Furthermore, the extracted solution may be concentrated to dryness, spray-dried, freeze-dried, etc., and used as a dried product.

In the present invention, the above extract may be used as is, or may contain ingredients such as fats and oils, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, metal soaps, pH adjusters, preservatives, fragrances, moisturizers, powders, UV absorbers, thickeners, pigments, antioxidants, whitening agents, chelating agents, excipients, coating agents, sweeteners, acidulants, etc., which are used in cosmetics, quasi-drugs, medicines, foods, etc., within the scope of the extract's effects.

The present invention can be used in any of cosmetics, quasi-drugs, medicines, and foods, and examples of the dosage forms include lotions, creams, milky lotions, gels, aerosols, essences, packs, cleaning agents, bath agents, foundations, dusting powders, lipsticks, ointments, poultices, tablets, capsules, chocolate, gum, candy, beverages, powders, granules, tablets, sugar-coated tablets, capsules, syrups, pills, suspensions, liquids, emulsions, suppositories, and injectable solutions.

When used externally, the content of the above extract used in the present invention is preferably 0.0001% by weight or more, and more preferably 0.001 to 10% by weight, calculated as solid matter. Furthermore, 0.01 to 5% by weight is most preferable. 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 see an increase in effect, and it is uneconomical.

When taken internally, the dosage varies depending on age, body weight, symptoms, therapeutic effect, administration method, treatment time, etc. Generally, the daily dosage per adult is preferably 5 mg or more, more preferably 10 mg to 5 g. Furthermore, 20 mg to 2 g is most preferable.

In the following, in order to explain the present invention in detail, manufacturing examples, experimental examples and formulation examples of the extract used in the present invention are given as examples, but the present invention is not limited to these. The percentages shown in the manufacturing examples are % by weight, and the parts of the content shown in the formulation examples are parts by weight.

Production Examples of Rowan Fruit Extracts Rowan fruit extracts were produced as follows: In Production Examples 1 to 4, Rowan fruit was used as the extraction material.

(Production Example 1) Preparation of hot water extract of rowan fruit 200 mL of water was added to 10 g of dried rowan fruit, and extraction was carried out for 2 hours at 95 to 100° C. The obtained extract was filtered, and the filtrate was concentrated and freeze-dried to obtain 3.6 g of hot water extract of rowan fruit.

(Production Example 2) Preparation of 50% ethanol extract of Rowan fruit 10 g of dried Rowan fruit was immersed in 200 mL of 50% ethanol solution at room temperature for 7 days to perform extraction. The obtained extract was filtered and then concentrated to dryness using an evaporator to obtain 3.5 g of 50% ethanol extract of Rowan fruit.

(Production Example 3) Preparation of ethanol extract of Rowan fruit 10 g of dried Rowan fruit was immersed in 200 mL of ethanol at room temperature for 7 days to perform extraction. The obtained extract was filtered and then concentrated to dryness using an evaporator to obtain 3.3 g of ethanol extract of Rowan fruit.

(Production Example 4) Preparation of 1,3-butylene glycol extract of rowan fruit 10 g of dried rowan fruit was immersed in 200 mL of 1,3-butylene glycol at low temperature for 7 days to perform extraction. The obtained extract was filtered to obtain 192 g of 1,3-butylene glycol extract of rowan fruit.

Next, experimental examples will be presented to explain the effects of the present invention in detail.

Experimental Example 1 Measurement of mRNA expression levels of type I collagen (COL1A1), hyaluronic acid synthase 2 (HAS2), MMP-1 and MMP-2 Human dermal fibroblasts (or keratinocytes) were seeded at 1×10 ⁵ cells on a 60 mm dish and cultured in DMEM culture medium containing 10% FBS under conditions of 37° C. and 5% CO _2. When the cells were confluent, they were cultured for 24 hours in DMEM(−) culture medium to which each sample was added so as to have a final concentration of 1, 10 or 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 amount of mRNA expression was measured by real-time RT-PCR 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, 500 ng of total RNA was subjected to reverse transcription reaction, followed by PCR reaction (95°C: 15 seconds, 60°C: 60 seconds, 40 cycles). Other operations were performed according to the prescribed method, and the expression levels of COL1A1, HAS2, MMP-1 and MMP-2 mRNA were calculated as a ratio to the expression level of GAPDH mRNA, which is an internal standard. The COL1A1 expression rate was calculated as the ratio of the expression level of COL1A1 mRNA in the sample-added group to the expression level of COL1A1 mRNA in the control (no sample added) group. The expression rates of HAS2, MMP-1 and MMP-2 were calculated in the same manner. The primers used for measuring the expression level of each gene are as follows.

Primer set for COL1A1 AGGACAAGAGGCATGTCTGGTT (SEQ ID NO: 1)
TTGCAGTGGTAGGTGATGTTCTG (SEQ ID NO: 2)
Primer set for HAS2: TGGATGACCTACGAAGCGATT (SEQ ID NO: 3)
GCTGGATTACTGTGGCAATGAG (SEQ ID NO: 4)
Primer set for MMP-1: GGGAGATCATCGGGGACAACTC (SEQ ID NO:5)
TGAGCATCCCCTCCAATACC (SEQ ID NO: 6)
Primer set for MMP-2 CCGTCGCCCATCATCAA (SEQ ID NO: 7)
CTTCTGCATCTTCTTTAGTGTGTCCTT (SEQ ID NO: 8)
Primer set for GAPDH: CACTCTTCCAGCCTTCCTTCC (SEQ ID NO: 9)
GTGTTGGCGTACAGGTCTTTG (SEQ ID NO: 10)

These experimental results are shown in Tables 1 to 4. As a result, the Rowan fruit extract of the present invention was found to have excellent COL1A1 expression promoting effect (collagen production promoting effect), HAS2 expression promoting effect (hyaluronic acid production promoting effect), MMP-1 expression suppressing effect (MMP inhibitory effect) and MMP-2 expression suppressing effect (MMP inhibitory effect). In particular, the Rowan fruit hot water extract (Production Example 1) was significantly effective in promoting COL1A1 expression and suppressing MMP-1 expression, the 50% ethanol extract (Production Example 2) was significantly effective in suppressing MMP-2 expression, and the ethanol extract (Production Example 3) was significantly effective in promoting COL1A1 expression and suppressing HAS2 expression.

Experimental Example 2: Melanin production inhibition test using B16 mouse melanoma B16 mouse melanoma was seeded in a 60 mm dish at 3×10 ⁴ cells, and cultured for 5 days in MEM culture medium containing 10% FBS added to each sample to a final concentration of 10 μg/mL) at 37° C. and 5% CO _2. After culture, the cells were detached, centrifuged, and the pellet obtained was dissolved in PBS (-) by ultrasonic disruption. Protein quantification was performed using the Lowry method (J. Biol. Chem., 193, 265-275, 1951). In addition, when measuring the amount of melanin, 4N NaOH was added to the remaining cell disruption solution taken for protein quantification, and the solution was heated at 60° C. for 2 hours, and then the absorbance at 475 nm was measured using a spectrophotometer (Shimadzu Corporation), the amount of melanin was obtained from the calibration curve, and the amount of melanin per 1 mg of protein was calculated. The melanin production inhibition rate was calculated from the ratio of the reduction in the amount of melanin in the sample-added group to the control (no sample-added) group.

These test results are shown in Table 5. It was confirmed that the hot water extract of Rowan fruit of the present invention (Production Example 1) has an excellent melanin production inhibitory effect.

Experimental Example 3: Active oxygen elimination test (antioxidation test)
The free radical scavenging and removal activity was evaluated. Ascorbic acid was used as a positive control. As a model of free radicals, a stable free radical, α,α-diphenyl-β-picrylhydrazyl (hereinafter referred to as DPPH), was used. It was reacted with the sample at a certain ratio for a certain period of time, and the amount of radicals that decreased was measured from the decrease in absorbance at a wavelength of 517 nm.

Measurement method of free radical scavenging and elimination action Each sample was added to 2 mL of 1.0 M acetate buffer (pH 5.5) to a final concentration of 100 μg/mL (ascorbic acid was 20 μg/mL), and 2 mL of anhydrous ethanol and 1 mL of 0.5 mM DPPH anhydrous ethanol solution were added to the reaction solution. In the case of an oil-soluble sample, the sample was added to 2 mL of anhydrous ethanol to prepare a reaction solution. After that, the reaction was carried out at 37° C. for 30 minutes, and the absorbance (A) at a wavelength of 517 nm was measured using water as a control. In addition, the absorbance (B) was measured using purified water instead of the sample as a blank. The free radical scavenging and elimination rate was calculated from the following formula.
Free radical scavenging and removal rate (%) = (1 - A/B) x 100

These test results are shown in Table 6. It was confirmed that the hot water extract (Production Example 1) and 50% ethanol extract (Production Example 2) of Rowan fruit of the present invention have a stable and excellent free radical scavenging and removal effect.

(Formulation Example 1) Lotion Formulation Content (%)
1. Hot water extract of Rowan fruit (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 parahydroxybenzoate 0.1
9. Polyoxyethylene hydrogenated castor oil (40 E.O.) 0.1
10. Fragrance (as needed) 11. Add purified water to make the total volume 100 [Manufacturing method] Dissolve ingredients 1-6 and 11 and ingredients 7-10 uniformly, mix the two, and filter to make the product.

Comparative Formulation Example 1: Conventional lotion A conventional lotion was prepared by replacing the hot water extract of Rowan fruit in Formulation Example 1 with purified water.

(Formulation Example 2) Cream Formulation Content (%)
1. 50% ethanol extract of Rowan fruit (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. Fragrance 0.1
11. Methyl parahydroxybenzoate 0.2
12. 1,3-butylene glycol 8.5
13. Make the total volume 100 with purified water. [Manufacturing method] Heat and dissolve ingredients 2-9, mix, and maintain at 70°C to make the oil phase. Heat and dissolve ingredients 1 and 11-13, mix, and maintain at 75°C to make the water phase. Add the water phase to the oil phase to emulsify, cool with stirring, add ingredient 10 at 45°C, and further cool to 30°C to make the product.

Comparative Formulation Example 2: Conventional Cream A conventional cream was prepared by replacing the 50% ethanol extract of Rowan fruit in Formulation Example 2 with purified water.

(Formulation Example 3) Emulsion Formulation Content (%)
1. Ethanol extract of Rowan fruit (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 (20 E.O.) 2.0
9. Fragrance 0.1
10. Propylene glycol 1.0
11. Glycerin 2.0
12. Methyl parahydroxybenzoate 0.2
13. Make the total volume 100 with purified water. [Manufacturing method] Heat and dissolve ingredients 1-8, mix, and keep at 70°C to make the oil phase. Heat and dissolve ingredients 10-13, mix, and keep at 75°C to make the water phase. Add the water phase to the oil phase to emulsify, cool with stirring, add ingredient 9 at 45°C, and cool further to 30°C to make the product.

(Formulation Example 4) Gel Formulation Content (%)
1. 1,3-butylene glycol extract of Rowan fruit (Production Example 4) 1.0
2. Ethanol 5.0
3. Methyl parahydroxybenzoate 0.1
4. Polyoxyethylene hydrogenated castor oil (60 E.O.) 0.1
5. Fragrance: appropriate amount 6. 1,3-butylene glycol 5.0
7. Glycerin 5.0
8. Xanthan gum 0.1
9. Carboxyvinyl polymer 0.2
10. Potassium hydroxide 0.2
11. Add purified water to make the total volume 100. [Manufacturing method] Dissolve ingredients 2 to 5, 1, and 6 to 11 uniformly, and mix the two to make the product.

(Formulation Example 5) Pack Formulation Content (%)
1. Hot water extract of Rowan fruit (Production Example 1) 1.0
2. 1,3-butylene glycol extract of Rowan fruit (Production Example 4) 5.0
3. Polyvinyl alcohol 12.0
4. Ethanol 5.0
5. 1,3-butylene glycol 8.0
6. Methyl parahydroxybenzoate 0.2
7. Polyoxyethylene hydrogenated castor oil (20 E.O.) 0.5
8. Citric acid 0.1
9. Sodium citrate 0.3
10. Fragrance (as needed) 11. Add purified water to make a total of 100 parts. [Manufacturing Method] Dissolve ingredients 1 to 11 uniformly to make the product.

(Formulation Example 6) Foundation Formulation Content (%)
1. 50% ethanol extract of Rowan fruit (Production Example 2) 1.0
2. Stearic acid 2.4
3. Polyoxyethylene sorbitan monostearate (20 E.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. Sodium carboxymethylcellulose 0.1
10. Bentonite 0.5
11. Propylene glycol 4.0
12. Triethanolamine 1.1
13. Methyl parahydroxybenzoate 0.2
14. Titanium dioxide 8.0
15. Talc 4.0
16. Bengala 1.0
17. Yellow iron oxide 2.0
18. Flavoring, appropriate amount 19. Make the total volume 100 with purified water [Manufacturing method] Heat and dissolve ingredients 2 to 8, and keep at 80°C to make the oil phase. Allow ingredient 9 to swell well in ingredient 19, then add ingredients 1 and 10 to 13 and mix uniformly. Add ingredients 14 to 17, which have been ground and mixed in a grinder, and stir with a homomixer while keeping the temperature at 75°C to make the water phase. Add the water phase to the oil phase while stirring, and emulsify. Then cool, add ingredient 18 at 45°C, and cool to 30°C while stirring to make the product.

(Formulation Example 7) Bath additive Formulation Content (%)
1. Ethanol extract of Rowan fruit (Production Example 3) 1.0
2. Sodium bicarbonate 50.0
3. Yellow No. 202 (1) appropriate amount 4. Fragrance appropriate amount 5. Sodium sulfate to make the total amount 100 [Manufacturing method] Mix ingredients 1 to 5 uniformly to make the product.

(Formulation Example 8) Ointment Formulation Content (%)
1. Hot water extract of Rowan fruit (Production Example 1) 5.0
2. 1,3-butylene glycol extract of Rowan fruit (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 parahydroxybenzoate 0.1
8. Propylene glycol 10.0
9. Make the total volume 100 with purified water. [Manufacturing method] Heat and dissolve ingredients 3-6, mix, and keep at 70°C to make the oil phase. Heat and dissolve ingredients 1, 2, and 7-9, mix, and keep at 75°C to make the water phase. Add the water phase to the oil phase to emulsify, and cool to 30°C while stirring to make the product.

(Formulation Example 9) Powder Formulation Content (%)
1. Hot water extract of Rowan fruit (Production Example 1) 1.0
2. Dried cornstarch 39.0
3. Microcrystalline cellulose 60.0
[Manufacturing method] Mix ingredients 1 to 3 to make a powder.

(Formulation Example 10) Tablet Formulation Content (%)
1. Ethanol extract of Rowan fruit (Production Example 3) 5.0
2. Dried 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. Component 6 is added to the formed granules and compressed into tablets. Each tablet weighs 0.52 g.

(Formulation Example 11) Tablets Formulation Content (%)
1. Ethanol extract of Rowan fruit (Production Example 3) 2.0
2. Dried cornstarch 49.8
3. Erythritol 40.0
4. Citric acid 5.0
5. Sucrose fatty acid ester 3.0
6. Fragrance 0.1
7. Add purified water to make up to 100%
[Manufacturing method] Mix ingredients 1 to 4 and 7 and mold into granules. Add ingredients 5 and 6 to the molded granules and compress into tablets. Each tablet weighs 1.0 g.

(Formulation Example 12) Beverage Formulation Content (%)
1. Hot water extract of Rowan fruit (Production Example 1) 0.05
2. Stevia 0.05
3. Malic acid 5.0
4. Fragrance 0.1
5. Add purified water to make the total volume 100. [Production method] Dissolve components 2 and 3 in a small amount of water. Next, add components 1, 4, and 5 and mix.

From the above, the Rowan fruit extract of the present invention has excellent collagen production promoting effects, hyaluronic acid production promoting effects, MMP inhibitory effects, melanin production suppression effects, and antioxidant effects, and is also highly stable. Therefore, the Rowan fruit extract 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 inhibiting functional decline due to aging, and preventing and treating cancer, and is expected to be applied to foods, cosmetics, quasi-drugs, pharmaceuticals, etc.

Claims (13)

An MMP-1 inhibitor characterized by containing an extract of Rowan fruit using water or using aqueous ethanol with an ethanol concentration of 50% by weight or less. An MMP-2 inhibitor characterized by containing an extract of Rowan fruit using water, aqueous ethanol or ethanol. A collagen production promoter (excluding those for improving wrinkles) characterized by containing extract of Rowan fruit. A hyaluronic acid production promoter (excluding those for improving wrinkles) characterized by containing extract of Rowan fruit. An antioxidant characterized by containing an extract of Rowan fruit in water at 95-100°C. A pharmaceutical for preventing and improving various diseases caused by the enhancement of MMP-1, comprising an extract of Rowan fruit with water or with aqueous ethanol having an ethanol concentration of 50% by weight or less. A pharmaceutical for preventing and improving various diseases caused by the enhancement of MMP-2, characterized by containing an extract of Rowan fruit using water, aqueous ethanol or ethanol. A pharmaceutical product for preventing and improving joint diseases caused by decreased collagen production and/or physiological floaters caused by decreased hyaluronic acid production, characterized by containing an extract of Rowan fruit. A medicine for preventing and improving various diseases caused by a decrease in the ability to scavenge active oxygen, characterized by containing an extract of Rowan fruit in water at 95-100°C. A food composition for preventing and improving various diseases caused by the enhancement of MMP-1, characterized by containing an extract of Rowan fruit with water or with aqueous ethanol having an ethanol concentration of 50% by weight or less. A food composition for preventing and improving various diseases caused by the enhancement of MMP-2, characterized by containing an extract of Rowan fruit with water, aqueous ethanol or ethanol. A food composition for preventing and improving joint diseases caused by decreased collagen production and/or physiological floaters caused by decreased hyaluronic acid production, characterized by containing an extract of Rowan fruit. A food composition for preventing and improving various diseases caused by a decrease in the ability to scavenge active oxygen, which comprises an extract of Rowan fruit with water at 95-100°C.