JP7148115B2 - Collagen production promoter, MMP inhibitor, melanogenesis inhibitor, cell proliferation promoter, antioxidant, wrinkle improving agent, pharmaceutical or food composition - Google Patents

Description

TECHNICAL FIELD The present invention relates to collagen production promoters, MMP inhibitors, melanin production inhibitors, cell proliferation promoters, antioxidants, anti-wrinkle agents, pharmaceuticals or food compositions.

Skin is exposed to various physical and chemical stresses such as ultraviolet rays, 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 Asclepix, tenki and xanthus xylophilus has been reported as an active ingredient for improving wrinkles caused by ultraviolet irradiation (Patent Document 2).

Fibroblasts and collagen are present in the dermis, and type I collagen accounts for 80% of the total. Besides type I collagen, the existence of type III, V, XII and XIV collagens 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.

Collagen is also a major structural protein, accounting for about one-third of mammalian tissues, and an essential component of many matrix tissues such as cartilage, bone, tendon, and skin. Collagen molecules, which are stable in normal tissues, are denatured into single-stranded gelatin when cleaved at one site by collagenase (MMP-1), which belongs to MMP, and become degraded by various other proteases. . As a result, the structural integrity of the matrix tissue is lost.

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.

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 believed that this connective tissue loses contractile force and further loses elasticity, resulting in wrinkles and sagging of the skin.

In general, skin pigmentation such as age spots, freckles, sunburn, etc. occurs when melanin pigment-producing cells in the skin overproduce melanin pigment due to hormonal abnormalities and ultraviolet stimulation, and this melanin pigment is deposited in the skin. is thought to be the cause. One known method for preventing such pigmentation is to suppress the excessive production of melanin. Conventionally, ascorbic acid (vitamin C) or the like has been used internally or externally for the treatment of pigmentation (Patent Document 6).

With aging, epidermal cell proliferation/division ability decreases, and the epidermal layer itself becomes thinner (Non-Patent Document 1). Biological factors such as epidermal growth factor (EGF/epidermal growth factor) and female hormones (estrogen) act on epidermal cell proliferation in the skin, but their secretion decreases with aging. Such deterioration in epidermal cell 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 ameliorate the progression of these phenomena, searches have been made for components that promote proliferation of epidermal cells, and many proposals have been made for topical skin preparations.

In addition, the skin is located in the outermost layer of the living body, is an organ that is likely to generate active oxygen under the influence of ultraviolet rays, etc., and is constantly exposed to oxygen stress. On the other hand, active oxygen scavenging enzymes are present in skin cells, and protect skin cells from injury by active oxygen unless the amount of active oxygen exceeding its capacity is generated. However, it is known that the activity of active oxygen scavenging enzymes in skin cells decreases with age. is thought to proceed. In addition, when organs other than the skin are exposed to active oxygen that exceeds their ability to scavenge active oxygen, it is thought that functional deterioration occurs and aging occurs, and various lifestyle-related diseases such as cancer and myocardial infarction develop. Therefore, active oxygen scavenging agents and antioxidants have been studied for the purpose of protecting against damage caused by active oxygen, and active oxygen scavenging enzymes such as SOD and catalase, and active oxygen scavenging agents and antioxidants such as SOD-like active substances are included. Foods, cosmetics, quasi-drugs, pharmaceuticals, etc. that have been developed have been developed (Patent Documents 7 and 8).

Borago officinalis, which belongs to the genus Borago of the family Boraginaceae of the Labiatae family, is characterized by the fact that its vegetable oil and seed oil have the effect of improving the symptoms of atopic dermatitis and are applied to skin external preparations (Patent Documents 9 and 10), and plants It is known that body extracts have a ceramidase activity inhibitory action and are applied to external preparations for skin (Patent Document 11), and have ameliorating effects on obesity and diabetes (Patent Document 12). However, it has not been known that these borages have collagen production promoting action, MMP inhibitory action, melanin production suppressing action, cell proliferation promoting action and antioxidant action.

JP-A-2000-119125 JP 2006-199611 A JP-A-3-44331 JP-A-2003-137801 JP-A-5-186368 JP-A-05-229931 JP-A-9-118630 JP-A-9-208484 JP-A-11-322628 JP-A-2002-53428 JP 2017-124984 A Japanese Patent Publication No. 2016-537422

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, melanogenesis-inhibiting action, cell growth-promoting action, and antioxidant action, but there is still no satisfactory material available. The current situation is that there is none.

Under these circumstances, the present inventors have made intensive studies and found that the borage extract has excellent collagen production promoting action, MMP inhibitory action, melanin production suppressing action, cell growth promoting action and antioxidant action, and is stable. It was found to be excellent also in Furthermore, the external preparation or internal preparation containing the extract is safe and stable, and has excellent collagen production promoting action, MMP inhibitory action, melanin production suppressing action, cell growth promoting action and antioxidant action, We have found that it can be used as a functional beauty/health material/medicine, and have completed the present invention.

Borage used in the present invention is a plant and a kind of herb, and is native to the Mediterranean coast of central Europe. It grows about 30-60 cm in height, has pale green leaves and stems covered with white hairs, and blooms blue or white star-shaped flowers in summer.

The extract of borage in the present invention is Borago officinalis (scientific name: Borago officinalis) belonging to the genus Borago of the family Boraginaceae of the Labiatae. or extracted from the whole plant. The extraction method is not particularly limited, and for example, it may be extracted with heating or extracted at room temperature. For extraction, the plant body may be used as it is, or may be subjected to a treatment such as drying, pulverization, or chopping.

The extraction method is not particularly limited, but it can be carried out by stirring or column extraction using water, hot water, or a mixed solvent of water and an organic solvent. 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. A particularly preferred extraction solvent includes water or a water-ethanol mixed polar solvent. The amount of the solvent to be used is not particularly limited. For example, it may be 10 times or more, preferably 20 times or more, relative to the whole plant (dry weight) of borage, but concentration or isolation after extraction may be performed. 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 and 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 within the range that does not impair the effect of the extract, oils and fats, waxes, and carbonized oils that are ingredients used in cosmetics, quasi-drugs, pharmaceuticals, foods, etc. Hydrogens, fatty acids, alcohols, esters, surfactants, metal soaps, pH adjusters, preservatives, fragrances, moisturizers, powders, UV absorbers, thickeners, pigments, antioxidants, whitening agents , chelating agents, excipients, film-forming agents, sweeteners, acidulants and the like.

The present invention can be used for any of cosmetics, quasi-drugs, pharmaceuticals, and foods. , bath agents, foundations, powders, lipsticks, ointments, poultices, tablets, capsules, chocolates, gums, candies, beverages, powders, granules, tablets, sugar-coated tablets, capsules, syrups, pills, suspensions preparations, liquids, emulsions, suppositories, solutions for injection 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. % shown in Production Examples means % by weight, and parts of content shown in Formulation Examples means parts by weight.

Preparation Example of Borage Extract An extract of borage was prepared as follows. Whole borage was used as an extraction material in Production Examples 1 to 4.

(Production Example 1) Preparation of hot water extract of borage To 10 g of dried borage, 200 mL of water was added and extracted at 95 to 100°C for 2 hours. The resulting extract was filtered, and the filtrate was concentrated and freeze-dried to obtain 1.4 g of a hot water extract of borage.

(Production Example 2) Preparation of 50% Ethanol Extract of Borage 10 g of dried borage was immersed in 200 mL of a 50% ethanol aqueous solution at room temperature for 7 days for extraction. After filtering the resulting extract, it was concentrated to dryness using an evaporator to obtain 1.1 g of a 50% ethanol extract of borage.

(Production Example 3) Preparation of ethanol extract of borage 10 g of dried borage was immersed in 200 mL of ethanol at room temperature for 7 days for extraction. After filtering the resulting extract, it was concentrated to dryness using an evaporator to obtain 0.4 g of an ethanol extract of borage.

(Production Example 4) Preparation of 1,3-butylene glycol extract of borage 10 g of dried borage was immersed in 200 mL of 1,3-butylene glycol at room temperature for 7 days for extraction. The resulting extract was filtered to obtain 192 g of a 1,3-butylene glycol extract of borage.

(Prescription example 1) Lotion Prescription Content (parts)
1. Hot water extract of borage (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 borage in Formulation Example 1 with purified water.

(Prescription example 2) Cream Prescription Content (parts)
1. 50% ethanol extract of borage (manufacturing 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 heated, melted 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 A conventional cream was obtained by replacing the 50% ethanol extract of borage in Formulation Example 2 with purified water.

(Prescription example 3) Milky lotion Prescription Content (parts)
1. Borage ethanol extract (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 (parts)
1. 1,3-butylene glycol extract of borage (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 the two are mixed to obtain a product.

(Prescription example 5) Pack prescription Content (parts)
1. Hot water extract of borage (Production Example 1) 1.0
2. 1,3-butylene glycol extract of borage (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 (parts)
1. 50% ethanol extract of borage (manufacturing 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 (parts)
1. Borage ethanol extract (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 (parts)
1. Hot water extract of borage (Production Example 1) 5.0
2. 1,3-butylene glycol extract of borage (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 (parts)
1. Hot water extract of borage (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 (parts)
1. Borage ethanol extract (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 tableted. One tablet is 0.52 g.

(Prescription example 11) Tablet prescription Content (parts)
1. Borage ethanol extract (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. Make the total amount 100 with purified water
[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 (parts)
1. Hot water extract of borage (Production Example 1) 0.05
2. Stevia 0.05
3. Malic acid 5.0
4. Perfume 0.1
5. The total amount is adjusted to 100 with purified water [Manufacturing method] Components 2 and 3 are dissolved in a small amount of water. Components 1, 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) and MMP-1 mRNA Expression Levels 1×10 ⁵ human fibroblasts NB1RGB were seeded on a φ60 mm dish, and when confluent, the final concentration was 1 or 10 μg/ Samples were added to make up to mL. Controls were added with the solvent in which the samples were diluted. After culturing for 24 hours, 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. High Capacity RNA-to-cDNA Kit (Applied Biosystems) and SYBR Select Master Mix (Life Technologies) were used for the real-time RT-PCR method. 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 the prescribed method, and the expression levels of COL1A1 and MMP-1 mRNA were determined as a ratio to the expression level of β-actin 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. The MMP-1 expression rate was also calculated in the same manner. 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 CACTCTTCCAGCCTTCCTTCC for β-Actin (SEQ ID NO: 5)
GTGTTGGGCGTACAGGTCTTTG (SEQ ID NO: 6)

The results of these experiments are shown in Tables 1-2. As a result, the borage extract of the present invention was found to have an excellent COL1A1 expression promotion effect (collagen production promotion effect) and MMP-1 expression suppression effect (MMP inhibition effect). In particular, the 50% ethanol extract of borage (Production Example 2) and the ethanol extract (Production Example 3) were remarkably effective in promoting COL1A1 expression and suppressing MMP-1 expression.

Experimental Example 2 Melanogenesis Suppression Test Using B16 Mouse Melanoma B16 mouse melanoma in logarithmic growth phase was seeded with 3×10 ⁴ cells on a φ60 mm dish, and each sample (final concentration 10 μg/mL) and 10% FBS were added. Eagle's MEM medium containing the cells was added and cultured for 5 days at 37° C., 5% CO ₂ conditions. After culturing, the cells were detached from the dish, sonicated, added with 4N NaOH and treated at 60° C. for 2 hours, and the absorbance at 475 nm was measured with a spectrophotometer. In addition, the cell lysate after sonication was quantified by Lowry's method (J.Biol.Chem., 193, 265-275, 1951), and the amount of melanin per protein amount was compared to suppress melanin production. It was used as an index of action.

These test results are shown in Table 3. It was confirmed that the borage hot water extract (Production Example 1), 50% ethanol extract (Production Example 2) and ethanol extract (Production Example 3) of the present invention have an excellent melanin production inhibitory effect. was taken.

Experimental Example ³ Cell proliferation promotion test HaCaT cells were seeded in a DMEM culture medium containing 0.1% FBS at 5 × 10 per well in a 96-well plate, and after adding each sample, the cells were incubated at 37°C and 5% CO. Cultured for 4 days under ₂ conditions. Cell count was measured by staining method. That is, after completion of the culture, 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 0.1 N HCl was added to each well and well stirred, and the absorbance at 650 nm was measured using a microplate reader.

These experimental results are shown in Table 4. As a result, the borage hot water extract (Production Example 1), the 50% ethanol extract (Production Example 2), and the ethanol extract (Production Example 3) of the present invention exhibited an excellent cell growth promoting effect.

Experimental Example 4 Active Oxygen Scavenging Action The free radical scavenging and removing action was evaluated. Ascorbic acid was used as a positive control. As a model of free radicals, α,α-diphenyl-β-picrylhydrazyl (hereinafter referred to as DPPH), which is a stable free radical, is used and reacted with a sample at a constant rate for a certain period of time to reduce the amount of radicals. was measured from the amount of decrease in absorbance at a wavelength of 517 nm.

Measurement method for free radical scavenging and removing action 1 mL of 5 mM DPPH absolute ethanol solution was added to prepare a reaction solution. In the case of an oil-soluble sample, the sample was added to 2 mL of absolute ethanol to prepare a reaction solution. After that, reaction was carried out at 37° C. for 30 minutes, and absorbance (A) at a wavelength of 517 nm was measured using water as a control. Also, as a blank, purified water was used in place of the sample to measure the absorbance (B). The free radical capture/removal rate was calculated from the formula shown below.
Free radical capture and removal rate (%) = (1-A/B) x 100

These test results are shown in Table 5. The borage hot water extract (Production Example 1), 50% ethanol extract (Production Example 2) and ethanol extract (Production Example 3) of the present invention have stable and excellent free radical scavenging and removing effects. was recognized.

Experimental Example 5 Use test Using the lotion of Formulation Example 1 and the conventional lotion of Comparative Formulation Example 1, a one-month use test was conducted on 5 people (27 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 external preparation for skin containing the extract of the present invention reduced wrinkles and sagging. During the test period, none of the subjects had skin troubles, and there were no problems with safety. Moreover, there was no problem with deterioration of prescription components.

As described above, the borage extract of the present invention has excellent collagen production-promoting action, MMP-inhibiting action, melanin production-inhibiting action, cell proliferation-promoting action and antioxidant action, and is also excellent in stability. Therefore, the borage 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 suppression of functional deterioration due to aging, prevention and treatment of cancer, etc. Foods, cosmetics, quasi-drugs It is also expected to be applied to pharmaceuticals and the like.

Claims (3)

Characterized by containing an extract of borage (not including seeds) extracted with one or more solvents selected from water, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol and 2-butanol Collagen production promoter. Characterized by containing an extract of borage (not including seeds) extracted with one or more solvents selected from water, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol and 2-butanol A cell proliferation promoting agent to be. Characterized by containing an extract of borage (not including seeds) extracted with one or more solvents selected from water, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol and 2-butanol A wrinkle improving agent.