JP7076062B2 - External skin agents, anti-glycation agents and collagenase inhibitors - Google Patents

Description

The present invention relates to an external skin preparation, an anti-glycation agent and a collagenase inhibitor containing fermented ginger as an active ingredient.

It is known that ginger family plants contain components such as gingerol and shogaol, and have various effects such as bactericidal action, antioxidant action, and blood circulation promotion.

Various studies have been conducted on this ginger family plant, and for example, a method for enriching shogaol by converting gingerol in the ginger family plant into shogaol has been proposed (see Patent Document 1). Further, it has been reported that heat-fermented ginger obtained by fermenting a ginger family plant and then heating it has an effect of reducing triglyceride in blood (see Patent Document 2).

By the way, in recent years, in the beauty industry and the medical industry, it has been found that saccharification affects beauty and health together with oxidation, and anti-glycation action is attracting attention.

For example, the above ginger family plants are known to have anti-glycation activity, but little further research has been done on the anti-glycation activity of ginger family plants. In Patent Document 2 describing heat-fermented ginger, it is investigated whether or not the produced heat-fermented ginger has an effect of suppressing an increase in blood glucose level which contributes to saccharification. It is reported that there is no such thing (see FIG. 1 of Patent Document 2).

In the beauty industry and the like, research has been conducted to protect and maintain collagen in the body, but little research has been conducted on the effects of ginger family plants on the maintenance of collagen.

Japanese Patent No. 5297294 Japanese Unexamined Patent Publication No. 2014-205629

An object of the present invention is to provide a skin external preparation having a high anti-glycation effect and a high collagenase inhibitory effect. Another object of the present invention is to provide an anti-glycation agent having a high anti-glycation activity and a collagenase inhibitor having a high collagenase inhibitory activity.

The present inventors have been paying attention to the efficacy of ginger family plants, and as a result of further diligent research on the efficacy, they have found that fermented ginger has a high anti-glycation effect, and have completed the present invention. At the same time, the present inventors have found that fermented ginger has a high collagenase inhibitory effect, and have completed the present invention.

That is, the present invention is characterized by containing a skin external preparation characterized by containing fermented ginger as an active ingredient, an anti-glycation agent characterized by containing fermented ginger as an active ingredient, and fermented ginger as an active ingredient. Concers about collagenase inhibitors.

According to the present invention, it is possible to provide a skin external preparation having a high anti-glycation action and a high collagenase inhibitory action, an anti-glycation agent having a high anti-glycation action, and a collagenase inhibitor having a high collagenase inhibitory action.

It is a graph which shows the AGEs production inhibition rate (saccharification reaction inhibition rate) by the fermented ginger-containing composition which concerns on Example 1. FIG. The horizontal axis shows the concentration of the test substance (mg / mL). It is a graph which shows the collagenase inhibition rate by the fermented ginger-containing composition which concerns on Example 1. FIG. The horizontal axis shows the concentration of the test substance (mg / mL).

The external skin preparation of the present invention is not particularly limited as long as it contains fermented ginger as an active ingredient, and the external skin preparation of the present invention has a high anti-glycation effect and a high collagenase inhibitory effect. Due to its high anti-glycation effect, it is possible to suppress the production and accumulation of AGEs, which reduces the amount of collagenase secreted, helps maintain the function of collagen, prevents dullness and darkening, and maintains the transparency of the skin. be able to. In addition, due to its high collagenase inhibitory action, it is possible to suppress the destruction of collagen fibers, maintain the elasticity of the skin, and eliminate the causes of wrinkles and sagging. Therefore, due to the action of both of these, the external skin preparation of the present invention has a particularly high skin cosmetic effect.

Further, the anti-glycation agent of the present invention is not particularly limited as long as it contains fermented ginger as an active ingredient, and the anti-glycation agent of the present invention has a high anti-glycation effect. Due to its high anti-glycation effect, it is possible to suppress the production and accumulation of AGEs, which reduces the amount of collagenase secreted, helps maintain the function of collagen, prevents dullness and darkening, and maintains the transparency of the skin. be able to. Further, by suppressing the saccharification reaction (Maillard reaction) of amino acids and proteins contained in the product, the stability of the product can be enhanced. Further, by suppressing the saccharification reaction, the color tone of the product can be stabilized.

Further, the collagenase inhibitor of the present invention is not particularly limited as long as it contains fermented ginger as an active ingredient, and the collagenase inhibitor of the present invention has a high collagenase inhibitory effect. Due to its high collagenase inhibitory action, it is possible to suppress the destruction of collagen fibers, maintain the elasticity of the skin, and eliminate the causes of wrinkles and sagging. In addition, by suppressing the destruction of collagen fibers, periodontal disease and gingival inflammation can be prevented and the health of the oral cavity can be maintained. Furthermore, by suppressing the destruction of collagen fibers, it is possible to prevent the aging of blood vessels and bone tissues and maintain the health of blood vessels and bone tissues. That is, the collagenase inhibitor of the present invention has actions such as inhibition of collagen decomposition, inhibition of skin dermis decomposition, and suppression of skin transepidermal water loss.

The raw material of fermented ginger, which is an active ingredient of the external skin preparation, anti-glycation agent and collagenase inhibitor of the present invention, is not particularly limited as long as it is a plant of the family Zingiberaceae, and for example, Sanshu ginger, yellow ginger, etc. Various ginger such as Kintoki ginger and Tananaka ginger can be mentioned. As the form of the ginger family plant raw material used in the present invention, the rhizome can be used as it is, or it can be used as a slice or a crushed product . Examples of the crushed product include powder, granules and the like . In the present invention, it is particularly preferable to use a dry powder.

The fermented ginger in the present invention is not particularly limited as long as it is a fermented ginger family plant (which has undergone a fermentation step) as a raw material, and the method for fermenting the ginger family plant raw material is self-fermentation. , Fermentation with bacterial cells and the like, but fermentation with bacterial cells is preferable. As the bacterial cells, microorganisms usually used for fermentation such as aspergillus, yeast, lactic acid bacteria, acetic acid bacteria, and Bacillus subtilis can be used, and among these, aspergillus is preferably used. The microorganism used in the present invention may be one kind or two or more kinds.

Examples of the aspergillus used in the present invention include black aspergillus, white aspergillus, yellow aspergillus, red aspergillus, and the like, and commercially available products can be preferably used. Specifically, Aspergillus aspergillus (Aspergillus awamori) (Aspergillus aspergillus), Aspergillus saitoi (Aspergillus aspergillus), Aspergillus nakazawai (Aspergillus nakazawa) (Aspergillus nakazawai) Aspergillus luchensis (black aspergillus), Aspergillus niger (black aspergillus), Aspergillus kawachi (Aspergillus kawachii) (white aspergillus) Examples of microorganisms belonging to the genus Aspergillus can be mentioned.

Examples of the yeast include Devalyomaises, Saccharomyces, Pichia, Kluyberomyces, Torraspora, Shubia, Brettanomices, Cryptococcus, Elemoterium, Issachenchia, Croquera, Lipomyses. , Metoschnikovia, Rhodtorua, Sizoccaromyce, Zigosaccharomyces, Candida. Examples of lactic acid bacteria include microorganisms belonging to the genus Streptococcus and Lactobacillus.

Examples of the method for allowing the ginger family plant material to act on the ginger family plant material include a method of spraying the ginger family plant material with the cell and its culture solution, a method of immersing or adding the ginger family plant material to the cell culture solution, and the like. As the culture solution of the bacterial cells, a culture solution cultured using a synthetic or natural medium and a fermented solution derived from a plant can be exemplified.

Fermentation with cells is carried out, for example, at a temperature of 5 to 70 ° C, preferably 10 to 40 ° C, more preferably 15 to 30 ° C, still more preferably less than 30 ° C for 1 to 30 days, preferably 3 to 14 days. It is preferable to do it.

Specifically, as a method for producing fermented ginger which is an active ingredient of the external skin preparation, antisaccharide agent and collagenase inhibitor of the present invention, for example, the condition of 30 to 90 ° C. after mixing a ginger family plant with a fermented liquid is used. Under the method described in Patent Documents 1 and 2 (long-term manufacturing method) in which aging is carried out over a long period of time such as 120 to 500 hours, a fermentation process for fermenting a plant material of the family Zingiberaceae, and a fermented product of the family Zingiberaceae. Examples thereof include a manufacturing method (short-time manufacturing method) including an addition step of adding a plant raw material and a heating step of heat-treating a fermented product to which a ginger family plant raw material is added at a temperature exceeding 100 ° C. under pressure. The long-term production method is a method of producing over a long period of time by heating at a relatively low temperature during or after fermentation. It is a method of performing heat treatment at a high temperature under conditions.

Here, the short-time manufacturing method will be described in detail.
The fermentation step is the same as the fermentation method described above. In the subsequent addition step, it is preferable to use the same one as the ginger family plant raw material used in the fermentation step, but those having different types and forms may be used. The amount of the ginger family plant raw material added is preferably 0.5 to 50 parts by mass, preferably 1 to 30 parts by mass, 1 part by mass with respect to 1 part by mass of the ginger family plant raw material used in the fermentation step. It is more preferably .5 to 15 parts by mass.

The heating step following the addition step is a step of heat-treating at a temperature exceeding 100 ° C. under pressure, and the treatment is carried out within a time that the fermentation of the ginger family plant raw material added in the addition step does not sufficiently proceed (start). ) Is preferable. Specifically, for example, it is preferable to carry out the treatment of the heating step within 12 hours after the treatment of the addition step, more preferably within 6 hours, further preferably within 3 hours, and within 1 hour. It is particularly preferable to do so.

As the heating conditions, the heating temperature is preferably 105 to 200 ° C, more preferably 105 to 180 ° C, further preferably 105 to 150 ° C, and particularly preferably 105 to 120 ° C. preferable. Further, the heating time is preferably 1 to 24 hours, more preferably 1 to 12 hours, still more preferably 1 to 10 hours. After the heating step, a drying treatment such as spray drying, freeze drying, or the like can be performed.

The external skin preparation of the present invention may be used by being applied to the skin, scalp, etc., and examples thereof include forms such as ointments, creams, gels, lotions, milky lotions, soaps, packs, and compresses. can.

Further, the anti-glycation agent of the present invention has an improved anti-glycation effect as compared with the unfermented one, and can be distinguished from other products as a product in that it is used for anti-glycation applications. For example, the main body, packaging, instruction manual, or advertised product of the product according to the anti-glycation agent of the present invention may indicate that it has functions such as anti-glycation and suppression of Maillard reaction.

In addition, the collagenase inhibitor of the present invention has an improved anti-glycation effect or collagenase inhibitory effect as compared with the unfermented one, and is distinguished from other products as a product in that it is used for collagenase inhibitory use. Anything that can be done, for example, collagenase inhibition, collagen decomposition inhibition, skin dermal decomposition inhibition, skin transdermal water evaporation amount in any of the main body, packaging, instruction manual, and advertisement of the product according to the collagenase inhibitor of the present invention. It can be mentioned that it has a function such as suppression.

Specific examples of the anti-glycation agent or collagenase inhibitor of the present invention include pharmaceutical products (including non-pharmaceutical products), cosmetics, foods for specified health use, foods with nutritional function, foods with functional claims, etc. Examples thereof include so-called health foods such as functional foods whose efficacy has been approved.

The anti-glycation agent or collagenase inhibitor of the present invention can be used for external use or oral use. The external preparation is not particularly limited as long as it is applied to the skin, scalp, etc., and its form is an ointment, a cream, a gel, a lotion, a milky lotion, a pack, a poultice, etc. Examples thereof include forms such as external skin preparations and injections.

When the anti-glycation agent or collagenase inhibitor of the present invention is used as an oral preparation, the form thereof is, for example, tablets, capsules, powders, granules, liquids, granules, rods, plates, blocks. Examples thereof include liquids, solids, rounds, pastes, creams, capsules, gels, chewables, sticks and the like. Among these, the forms of tablets, capsules, powders, granules, and liquids are particularly preferable. Specifically, instant powder (granule) for dissolving in supplements, food additives, PET bottles, cans, bottled beverages filled in bottles, water (hot water), milk, fruit juice, green juice, etc. Beverages and the like can be exemplified. These are preferable in that they are easy to eat and drink at the time of meals and can enhance the palatability.

Further, the anti-glycation agent of the present invention can also be used as an additive (stabilizer for products) of products such as pharmaceuticals, cosmetics and health foods. The anti-glycation agent of the present invention can improve the stability of the product, for example, by suppressing the saccharification reaction (Maillard reaction) of the components in the product.

Hereinafter, the present invention will be described based on examples.

[Example 1]
In a GE medium (liquid medium) containing glucose and yeast extract, A. black aspergillus is used. Awamori was added and the cells were cultured at 28 ° C. for 2 days in a liquid culture tank. Then, it was diluted with GE medium so that the concentration of the cultured medium was 3.8%, and dry ginger powder (domestic ginger powder) was added so that the concentration was 1%. The medium to which the dry powder of ginger was added was cultured at 28 ° C. for 6 days in a culture tank for liquid, and the dry powder of ginger was fermented (fermentation step). After fermentation, the same dry ginger powder used in the fermentation step was added to a concentration of 3% (addition step). Immediately after the addition, heat treatment was performed at 115 to 120 ° C. for 2 hours under pressurized conditions (heating step). Then, it was dried by freeze-drying (drying step) to obtain a fermented ginger-containing composition (anti-glycation agent, collagenase inhibitor) according to Example 1 of the present invention.

Using the fermented ginger-containing composition according to Example 1, an anti-glycation confirmation test and a collagenase inhibition confirmation test were performed.

[Anti-glycation confirmation test]
This anti-glycation confirmation test is "Inhibitory effect of the compounds isolated from Rhus verniciflua on aldose reductase and advanced glycation end products." (Lee EH, Song DG, Lee JY, Pan CH, Um BH, Jung SH. Biol Pharm Bull. 2008 The method described in Aug; 31 (8): 1626-30.) Was slightly modified. Specifically, the procedure was as follows.

(Preparation of reagents)
Phosphate buffer powder (1/15 mol / L pH 7.2) (Wako Pure Chemical Industries, Ltd.) was dissolved in distilled water to prepare a 67 mM phosphate buffer (hereinafter abbreviated as 67 mM PB). In addition, D (+) glucose (Nacalai Tesque, Inc.) was dissolved in 67 mM PB to prepare a 200 mg / mL glucose solution. Further, albumin (bovine serum-derived corn fraction V, pH 7.0, for biochemistry) (Wako Pure Chemical Industries, Ltd.) (hereinafter abbreviated as BSA) was dissolved in 67 mM PB to prepare 40 mg / mL BSA. did.

(test)
A predetermined amount of the test substance (fermented ginger-containing composition according to Example 1 or ginger dry powder according to Comparative Example) was dissolved in 67 mM PB to prepare a test substance solution. The test substance solution, the glucose solution and the BSA solution were mixed at the ratios shown in Table 1 below to prepare a test solution (test). Similarly, a test solution (blank), a control (test), and a control (blank) were prepared by blending in the following ratios. The final concentration of the test substance is as shown on the horizontal axis (mg / mL) in FIG.

The test solution (test, blank) and control (test, blank) were incubated at 60 ° C. for 48 hours. For the test solution (test, blank) and control (test, blank) after incubation, the fluorescence intensity at 440 nm when excited at 370 nm was measured with a spectrofluorometer, and the AGEs production inhibition rate (%) was calculated by the following formula. Calculated.

Inhibition rate of AGEs production (%)
= (1-[(Sample test --Sample blank) / (Control test --Control blank)]) × 100

(In the formula, "Sample test" indicates the fluorescence intensity of the test solution (test), "Sample blank" indicates the fluorescence intensity of the test solution (blank), and "Control test" indicates the fluorescence intensity of the control (test). Indicates the intensity, and "Control blank" indicates the fluorescence intensity of the control (blank).)

The results are shown in FIG. As shown in FIG. 1, when the fermented ginger-containing composition according to Example 1 of the present invention was contained, the production of AGEs was clearly inhibited as compared with the case where the dry powder of ginger according to Comparative Example was contained.

[Collagenase inhibition confirmation test]
(Preparation of sample liquid)
Using Dalveco PBS (Phosphate Buffered Saline) (-) containing 0.1% BSA (bovine serum albumin), the test substance (fermented ginger-containing composition according to Example 1 or dry powder of ginger according to Comparative Example) was used. It was adjusted to 20 mg / mL. After shaking for 60 minutes with a vortex mixer, centrifuge to collect the supernatant, and further dilute the test substance with 0.1% BSA-containing Dalveco PBS (-) to a predetermined concentration to prepare a test solution. did. The final concentration of the test substance was prepared so as to be the value shown on the horizontal axis (μg / mL) in FIG.

(Preparation of enzyme solution)
Collagenase B (manufactured by Roche) was prepared to 10 μg / mL using Dulbecco PBS (−) containing 0.1% BSA.

(Preparation of fluorescent substrate solution)
Fluorescent substrate dissolved in DMSO (MOCAc-Pro-Leu-Gly-Leu-A ₂ pr (DNP) -Ala-Arg-NH ₂ , manufactured by Peptide Institute) was prepared with Dalveco PBS (-) containing 0.1% BSA. The solution diluted to 5 μM was used as a fluorescent substrate solution.

(Measurement of collagenase inhibitory activity)
Dulbecco PBS (-) containing 0.1% BSA as a test solution or control was added to 96 well black plate at 50 μL / well. Then, an enzyme solution (test) or Dulbecco PBS (−) (blank) containing 0.1% BSA was added at 100 μL / well and incubated at 37 ° C. for 10 minutes. Further, a fluorescent substrate solution was added at 50 μL / well, and the mixture was incubated at 37 ° C. for 60 minutes in the dark. It was excited at 320 nm and the fluorescence intensity at 405 nm was measured.

The collagenase inhibition rate of each test substance was calculated by the following formula.
Collagenase inhibition rate = (1-[(Sample test --Sample blank) / (Control test --Control blank)]) × 100

(In the formula, "Sample test" indicates the fluorescence intensity of the test solution (test), "Sample blank" indicates the fluorescence intensity of the test solution (blank), and "Control test" indicates the fluorescence intensity of the control (test). Indicates the intensity, and "Control blank" indicates the fluorescence intensity of the control (blank).)

The results are shown in FIG. As shown in FIG. 2, when the fermented ginger-containing composition according to Example 1 of the present invention was contained, the collagenase activity was clearly inhibited as compared with the case where the dry powder of ginger according to Comparative Example was contained.

(Formulation example 1: Toner)
With 100 parts by mass as a whole, 0.01 part by mass of fermented ginger extract, 0.01 part by mass of ginger extract, 10 parts by mass of glycerin, 3 parts by mass of diglycerin, 12 parts by mass of 1,3-butylene glycol, pentylene glycol. 3 parts by mass, 0.1 part by mass of sodium hyaluronate, 0.01 part by mass of citric acid, 0.02 part by mass of sodium citrate, 0.1 part by mass of xanthan gum, 0.15 part by mass of methylparaben, 0.2 part by mass of carbomer. , 0.03 parts by mass of sodium hydroxide and the water residue were mixed to prepare the composition of the present invention in the form of cosmetic water.

(Formulation example 2: shampoo)
With 100 parts by mass as a whole, 0.01 part by mass of fermented ginger extract, 0.02 part by mass of aloe extract, 7.5 parts by mass of sodium laureth sulfate, 4.2 parts by mass of cocamidopropyl betaine, 3 parts by mass of cocamide DEA. , 1,3-butylene glycol 0.1 part by mass, polyquaternium-10 0.225 part by mass, citric acid 0.15 part by mass, sodium citrate 0.05 part by mass, phenoxyethanol 0.9 part by mass and water residue mixed Then, the composition of the present invention was prepared in the form of shampoo.

(Formulation example 3: Soap)
With 100 parts by mass as a whole, 0.5 parts by mass of crushed fermented ginger, 2 parts by mass of glycerin, 1 part by mass of olive oil, 0.1 parts by mass of EDTA-4 sodium, 0.2 parts by mass of ethidronate 4 sodium and soap base. The composition of the invention was prepared in the form of soap by mixing and solidifying the balance.

(Formulation Example 4: Emulsion)
With 100 parts by mass as a whole, 0.1 part by mass of fermented ginger extract, 0.1 part by mass of thiamine hydrochloride, 3 parts by mass of sucrose fatty acid ester, 12 parts by mass of glycerin, 6 parts by mass of squalane, 24 parts by mass of dimethyl silicone oil. , 1 part by mass of polypropylene glycol, 0.06 part by mass of thickener, 0.2 part by mass of phenoxyethanol, 5 parts by mass of ethanol, 0.01 part by mass of sodium hydroxide and the balance of purified water are mixed to form a milky lotion. The composition of the invention was prepared.

(Formulation Example 5: Cosmetic cream)
With 100 parts by mass as a whole, 0.1 part by mass of fermented ginger extract, 0.1 part by mass of rosemary extract, 15.0 parts by mass of squalane, 4.0 parts by mass of octyldodecyl myristate, hydrogenated soybean phospholipid 0. 2 parts by mass, 2.4 parts by mass of butyl alcohol, 1.5 parts by mass of hardened oil, 1.5 parts by mass of stearate, 1.5 parts by mass of glycerin monostearate, which is a pro-oil type, 0.5 parts by mass of polyglyceryl monostearate. , Behenyl alcohol 0.8 parts by mass, polyglyceryl monomyristate 0.7 parts by mass, Sarashimitsurou 0.3 parts by mass, d-δ-tocopherol 0.1 parts by mass, methylparaben 0.3 parts by mass, C10-30 alkyl modified carboxy Mix 0.2 parts by mass of vinyl polymer, 0.1 parts by mass of carboxyvinyl polymer, 18.0 parts by mass of 1,3-butanediol, 0.1 parts by mass of sodium hydroxide and the balance of purified water to make a cosmetic cream. The composition of the present invention was prepared in an embodiment.

(Formulation example 6: Packing agent)
With 100 parts by mass as a whole, 0.1 part by mass of fermented ginger extract, 0.01 part by mass of soybean extract, 20.0 parts by mass of polyvinyl alcohol, 5.0 parts by mass of glycerin, 20.0 parts by mass of ethanol, 6.0 parts by mass of kaolin. The composition of the present invention was prepared in the form of a packing agent by mixing 0.2 parts by mass of a preservative, 0.1 parts by mass of a fragrance and a residue of purified water.

(Formulation Example 7: Tablet)
With 100 parts by mass as a whole, 10 parts by mass of fermented ginger powder, 8 parts by mass of karin powder, 5 parts by mass of vitamin B, 20 parts by mass of crystalline cellulose, 50 parts by mass of lactose, 4 parts by mass of magnesium stearate and the rest of corn starch are mixed and mixed. By tableting, the composition of the present invention was prepared in the form of tablets.

(Formulation Example 8: Granules)
The present invention is in the form of granules by mixing and granulating 10 parts by mass of fermented ginger powder, 15 parts by mass of lactose, 10 parts by mass of lactose, 1 part by mass of calcium stearate and the balance of crystalline cellulose, with the whole as 100 parts by mass. The composition of was prepared.

(Formulation Example 9: Capsule)
By encapsulating a mixture of 10 parts by mass of fermented ginger extract, 20 parts by mass of ginger extract, 8 parts by mass of lecithin and the balance of olive oil as the content liquid, with the whole as 100 parts by mass. , The composition of the present invention was prepared in the form of a capsule.

(Formulation Example 10: Liquid)
With 100 parts by mass as a whole, 0.84 parts by mass of fermented ginger extract powder, 1 part by mass of vitamin B12, 10 parts by mass of fructose-glucose liquid sugar, 1 part by mass of citric acid, 0.02 parts by mass of sodium benzoate, 2 parts by mass of fragrance preparation. The composition of the present invention was prepared in the form of a liquid preparation by mixing parts, 0.05 parts by mass of sucralose, 0.03 parts by mass of acesulfam potassium, and the balance of purified water.

Claims (1)

A cosmetic composition characterized by containing fermented ginger as an active ingredient, which is obtained by fermenting the rhizome of the ginger of the family Zingiberaceae, a slice of the rhizome, or a crushed product of the ginger with aspergillus. (Excluding cosmetics containing products obtained by adding plants and allowing ginger to act, as well as hair care products) .

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