JP2016056103A - Polyphenol derivative exhibiting keratin increasing activity and method for producing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a polyphenol derivative exhibiting keratin increasing activity and a method for producing the same.SOLUTION: The target polyphenol derivative is a derivative formed from one molecule each of polyphenol, arginine, and adenine. This derivative enhances production of keratin by activating a keratin-synthesizing enzyme. Keratin production is derived from two mechanisms including an enzyme activation mode and enzyme induction. The production method of the derivative comprises a step of protease treatment of adding a branched cyclodextrin to a fermentation liquid obtained by fermenting placenta and soybeans by Bacillus subtilis natto. The polyphenol derivative obtained can be applied to health food and cosmetics for skin beautifying effects. In addition, it can also be used in a purified medicinal drug and quasi drug which exhibit keratin increasing activity.SELECTED DRAWING: None

Description

The present invention relates to a polyphenol derivative exhibiting an action of increasing keratin and a method for producing the same.

Keratin is an extracellular protein that forms tissue with cells of epithelial tissue, and is present in hair and skin stratum corneum. It is characterized by containing a lot of cysteine as a constituent amino acid. Keratin is also involved in cytoplasmic flow in the cytoplasm, carbohydrate degrading enzymes, and fat accumulation.

Furthermore, keratin is abundant in skin epidermis cells and visceral epithelial cells, and is also involved in the regulation of the intracellular signal transduction system. In particular, it is involved in the control of tyrosine kinase and protein kinase functions.

This keratin decreases with age and is also associated with skin firmness and delayed skin regeneration. There are several types of keratin, and particularly keratin contained in the epidermis cells of the skin is abundant, and keratin is attracting attention and researched in the field of beauty and dermatology.

As an invention related to keratin, there is an invention of a cosmetic composition that forms a supramolecular polymer after application, and here, a chemical substance exhibiting a keratin action has been proposed. However, since this invention is a chemical substance, there is no description about a side effect (for example, refer patent document 1).

There is also an invention relating to the use of emulsified keratin used in cosmetics (see, for example, Patent Document 2). Here, although the use of cosmetics for keratin skin tissue is described, the technique relating to the increase of keratin is mild.

There is also an invention of a novel composition based on a joint mixture of at least one VDR ligand and a retinoid (see, for example, Patent Document 3). Here, a technique using keratin cells is described, but it is not a method of increasing keratin itself.

However, there are no inventions relating to methods, ingredients or substances for increasing keratin in the medical area, skin area or cosmetic area.

Thus, when the substance which increases keratin is a chemical substance, there exists a danger that a side effect will occur and industrial use is limited.

Therefore, a substance derived from a natural product and increasing keratin is desired.

Patent No. 3951000 Patent No. 5269287 Patent No. 2898100

As described above, the effect of increasing keratin by existing natural products is mild, and there is a problem that the industrial use is limited. Also, there is a problem in safety with chemically synthesized substances, and the use is limited. ing.

Therefore, a natural product that exhibits an excellent keratin increasing effect with weak side effects and a production method for efficiently producing the natural product are desired.

In order to achieve the above object, the invention described in claim 1 relates to a polyphenol derivative exhibiting a keratin increasing action represented by the following formula (1).

In order to achieve the above object, the invention described in claim 2 relates to a method for producing a polyphenol derivative exhibiting an action of increasing keratin.

Since this invention is comprised as mentioned above, there exist the following effects.

According to the derivative according to claim 1, it is possible to exhibit an excellent keratin increasing action.

According to the manufacturing method of Claim 2, the polyphenol derivative which exhibits a keratin increase effect | action efficiently can be manufactured.

Hereinafter, embodiments embodying the present invention will be described in detail.

First, the polyphenol derivative exhibiting the keratin increasing action represented by the following formula (1) is composed of one molecule of polyphenol, one molecule of polyphenol, and one molecule of cysteine.

The polyphenol part of this derivative is an isoflavone molecule and contains two oxygen elements in the molecule.

The carboxyl group of arginine is bonded to the benzene ring in the side chain of the polyphenol molecule. This arginine is L-type. The guanidine side chain and amino group of arginine exist as they are.

The guanidine side chain of arginine is preferred because it has the ability to produce nitric oxide. That is, endogenous nitric oxide synthase produces nitric oxide from arginine.

The produced nitric oxide acts on the smooth muscle cells of the blood vessels and relaxes, thereby expanding the blood vessels. This increases blood flow and lowers blood pressure. It is also preferable to increase skin blood flow and increase waste excretion and nutrient supply.

In addition, adenine is bonded to the mother nucleus of polyphenol. This is a bond with the 9th-position nitrogen element of adenine.

This polyphenol derivative can pass through the cell membrane and reach the cell because the mother nucleus is hydrophobic. Moreover, since it has a hydroxyl group, it is also water-soluble, and since it exhibits amphiphilic properties, it is preferable because it directly permeates the water-soluble keratin synthase after penetrating into the cell.

This polyphenol derivative activates the active center of keratin synthase to activate keratin synthesis. The mode of activation is reversible and its activation is stopped by the absence of the polyphenol derivative.

It is preferable from the viewpoint of safety that the activation of keratin synthase is reversible. That is, if this polyphenol derivative is decomposed, its function stops.

Since the polyphenol derivative contains polyphenol and has a hydroxyl group, production of active oxygen due to ultraviolet rays can be suppressed. Since ultraviolet rays and active oxygen cut keratin, it is preferable that this active oxygen can be removed.

This polyphenol derivative is preferable because it easily penetrates into skin cells and cells in subcutaneous tissue. That is, having an amphiphilic property is a preferable property.

Polyphenols originally have the function of regulating enzyme reactions. Inhibiting, activating, and allosteric effects are achieved by adjusting the catalytic reaction of the enzyme.

The polyphenol derivative shown here has a function of activating keratin synthase. The feature of the action is reversible, and the activation type is antagonistic. As an antagonistic feature, the work is temporary and reversible, so there is an advantage that side effects are unlikely to occur.

Keratin synthase is an enzyme distributed in tissues and cells throughout the body, and is present in liver, adipose tissue, blood, and the like.

As a method for extracting or producing this polyphenol derivative, any method such as a fermentation method, an enzyme reaction method or a chemical synthesis method is used.

The polyphenol derivative can be extracted from soybeans or placenta. In this extraction method, it is preferable to use digestive enzymes such as protease and lipase because the extraction efficiency is enhanced.

Moreover, in the case of an enzyme reaction method, it can extract from the plant containing a polyphenol. Alternatively, it can be biosynthesized by microorganisms by fermentation.

Furthermore, as a purification method, it is preferable to use a purification operation such as a separation resin.

For example, it is preferably separated and separated by a separation carrier or resin. As the separation carrier or resin, porous polysaccharides, silicon oxide compounds, polyacrylamide, polystyrene, polypropylene, styrene-vinylbenzene copolymers, etc., whose surfaces are coated as described later, are used. Those having a particle size of 0.1 to 300 μm are preferred. The finer the particle size, the higher the accuracy of the separation, but the longer the separation time.

For example, a reverse phase carrier or resin whose surface is coated with a hydrophobic compound is used for separation of a highly hydrophobic substance. Those coated with a cationic substance are suitable for the separation of anionically charged substances. Also, those coated with an anionic substance are suitable for separating a cationically charged substance. When a specific antibody is coated, it is used as an affinity carrier or resin for separating only a specific substance.

The affinity carrier or resin is used for specific preparation of an antigen using an antigen-antibody reaction. A partitionable carrier or resin is used for isolation of a substance such as silica gel (manufactured by Merck) if there is a difference in partition coefficient between the substance and the solvent for separation.

Among these, an adsorbent carrier or resin, a dispersible carrier or resin, a molecular sieve carrier or resin, and an ion exchange carrier or resin are preferable from the viewpoint of reducing production costs. Furthermore, the reverse phase carrier or resin and the dispersible carrier or resin are more preferable because the difference in the distribution coefficient with respect to the separation solvent is large.

When an organic solvent is used as the separation solvent, a carrier or resin having resistance to the organic solvent is used. Moreover, the carrier or resin used for pharmaceutical manufacture or food manufacture is preferable. From these points, Diaion (Mitsubishi Chemical Co., Ltd.) and XAD-2 or XAD-4 (Rohm and Haas) are used as the adsorptive carrier, and Sephadex LH-20 (Amersham Pharmacia) is used as the molecular sieve carrier. Silica gel as the distribution carrier, IRA-410 (Rohm and Haas) as the ion exchange carrier, and DM1020T (Fuji Silysia) as the reverse phase carrier are more preferable.

Of these, Diaion, Sephadex LH-20 and DM1020T are more preferred.

The obtained extract is dissolved in a solvent for swelling the carrier for separation or the resin before separation. The amount thereof is preferably 1 to 40 times the weight of the extract from the viewpoint of separation efficiency, and more preferably 4 to 20 times. The separation temperature is preferably 4 to 30 ° C., more preferably 10 to 25 ° C. from the viewpoint of the stability of the substance.

As the separation solvent, water or a lower alcohol containing water, a hydrophilic solvent, or a lipophilic solvent is used. As the lower alcohol, methanol, ethanol, propanol and butanol are used, and ethanol used for food is preferable.

When Sephadex LH-20 is used, a lower alcohol is preferable as the separation solvent. When silica gel is used, the separation solvent is preferably chloroform, methanol, acetic acid or a mixture thereof.

When Diaion and DM1020T are used, the separation solvent is preferably a lower alcohol such as methanol or ethanol or a mixed solution of lower alcohol and water.

In addition, it is preferable to collect a fraction containing the activity and remove the solvent by drying or vacuum drying to obtain a powder or a concentrated liquid because the influence of the solvent can be excluded.

This polyphenol derivative directly acts on cells existing in human skin and skin cells to biosynthesize keratin, and exhibits skin beautification and skin regeneration.

It is preferable to add fats and oils to the polyphenol derivative because the resulting active part is stably maintained in the oil. For example, extraction with soybean oil, rice bran oil, grape seed oil, olive oil or jojoba oil is preferred.

It is used as an injectable or parenteral agent such as an oral agent or a coating agent as a pharmaceutical, and as a quasi-drug, it is used in a tablet, capsule, drink, soap, coating agent, gel, toothpaste, etc. The

Examples of oral preparations include tablets, capsules, powders, syrups, and drinks. When mixed with the above-mentioned tablets and capsules, it can be used together with a binder, excipient, swelling agent, lubricant, sweetener, flavoring agent and the like. The tablets can also be coated with shellac or sugar.

Moreover, in the case of the said capsule, liquid carriers, such as fats and oils, can be further contained in said material. In the case of the above syrup and drink, sweeteners, preservatives, pigment flavoring agents and the like can be added.

Examples of parenteral preparations include injections in addition to external preparations such as ointments, creams, and liquids. Vaseline, paraffin, fats and oils, lanolin, macro gold, etc. are used as a base material for external preparations, and can be made into ointments, creams, and the like by ordinary methods.

Injections include liquids, and other lyophilization agents. This is used aseptically dissolved in distilled water for injection or physiological saline at the time of use.

It is used as a food preparation for beauty foods intended for beauty, foods intended for beauty, and cell activators intended to maintain healthy cells. Moreover, it is preferable to use as a health functional food for a nutritional functional food or a food for specified health use.

When the obtained food preparation is used for pets and livestock animals such as dogs and cats, it is used as a feed or a supplement for pets for the purpose of maintaining the health of the skin and tissues.

As a cosmetic, it can be used together with a surfactant, a solvent, a thickener, an excipient and the like according to a conventional method. For example, it can be in the form of cream, gel for hair, facial cleanser, beauty essence, lotion, etc., and becomes a cosmetic that promotes the production of keratin.

The form of the cosmetic is arbitrary, and can be used as a solution, cream, paste, gel, gel, solid or powder.

Next, the manufacturing method of the polyphenol derivative which exhibits the keratin increase effect | action which consists of the process of adding a placenta and soybean and fermenting fermented by natto bacteria and adding a branched cyclodextrin and performing protease treatment is demonstrated.

The polyphenol derivative here comprises one molecule of polyphenol, one molecule of arginine and one molecule of adenine.

The polyphenol part of this derivative is an isoflavone molecule and contains two oxygen molecules.

The carboxyl group of arginine is bonded to the benzene ring in the side chain of the polyphenol molecule. This arginine is L-type. The guanidine side chain and amino group of arginine exist as they are.

The guanidine side chain of arginine is preferred because it has the ability to produce nitric oxide. That is, nitric oxide is produced by the action of endogenous nitric oxide synthase.

Nitric oxide acts on the smooth muscle cells of blood vessels and relaxes them, thereby dilating the blood vessels. This increases blood flow and lowers blood pressure. It is also preferable to increase skin blood flow and increase waste excretion and nutrient supply.

In addition, adenine is bonded to the mother nucleus of polyphenol. This is a bond with the 9th-position nitrogen element of adenine.

This production method comprises a step of adding a placenta and soybeans, adding a branched cyclodextrin to a fermentation broth fermented with Bacillus natto, and subjecting it to protease treatment.

The raw materials are placenta, soybean, natto, branched cyclodextrin and protease.

The placenta is the placenta of animals, and the placenta of edible animals such as pigs, cows, horses, and sheep is preferable because of its high quality.

This country of origin can be used in Japan, Asia, America, Europe, etc. In particular, placenta in Japan is preferable because of its good quality and excellent bonding with polyphenols.

The soybean used as a raw material can be any soybean from Japan, China, the United States, Russia, etc., but it is preferable that the soybean is traceable and the producer is clear.

Of these, soybeans cultivated organically or without agricultural chemicals are more preferred because they do not contain harmful agricultural chemicals or metals.

When using soybeans, Nara Machinery Co., Ltd. free mill, super free mill, sample mill, goblin, super clean mill, micros, small vacuum dryer manufactured by Toyo Riko as vacuum dryer, small size manufactured by Matsui Co., Ltd. It is pulverized by a pulverizer such as a vacuum heat transfer dryer DPTH-40, clean dry VD-7, VD-20 manufactured by AKM Kyushu Technos Co., Ltd., DM-6 manufactured by Nakayama Technical Research Institute. Thereby, the process of fermentation tends to advance efficiently.

The Bacillus natto used is the scientific name Bacillus subtilis, which is a useful microorganism used in the production of natto. Natto bacteria, a powder of Natto Motopo, is preferable because of its good quality and suitable for fermentation.

A branched cyclodextrin is one of cyclic glucose, and is preferable because glucose is bound cyclically and used in foods and cosmetics. Since this branched cyclodextrin has a hydrophobic portion in the lumen, it is easy to adsorb highly hydrophobic substances. Branched cyclodextrins manufactured by Shimizu Minato Sugar Co., Ltd. are preferred because of their high quality.

As the protease to be used, the quality of protease A “Amano” SD, protease M “Amano” SD or protease P “Amano” 3SD, which are food processing proteases manufactured by Amano Enzyme, is preferable because of its stable use.

First, the placenta is pulverized by a pulverizer or a mixer. By being pulverized, it becomes easy to process. The soybeans are pulverized by a pulverizer and suspended by adding clean water. 50 to 300 g of soybean is added to 100 g of placenta, and stirred with water such as purified water and 1 to 10 liters in a clean container.

Placenta and soybeans are sterilized by boiling and added to the fermentation tank. By sterilization, contamination with various bacteria is prevented, and fermentation with Bacillus natto proceeds.

Fermentation may be either a stationary method or a stirring method, but a stirring method is preferred because fermentation can be performed in a short time. Fermentation is preferably performed at 39 to 45 ° C. for 23 to 71 hours. When the temperature is low and the time is short, fermentation does not proceed, and when the temperature is high and the time is long, the target polyphenol derivative may be decomposed.

The fermentation broth is preferably filtered with a filter cloth or the like because the following steps can be easily performed.

Branched cyclodextrin is added to the filtrate. A branched cyclodextrin is one of cyclic glucose, and is preferable because glucose is bound cyclically and used in foods and cosmetics. Since this branched cyclodextrin has a hydrophobic portion in the lumen, it is easy to adsorb highly hydrophobic substances. Branched cyclodextrins manufactured by Shimizu Minato Sugar Co., Ltd. are preferred because of their high quality.

The added branched cyclodextrin is preferably 40 to 400 g of branched cyclodextrin with respect to 100 g of placenta. With this branched cyclodextrin, placenta peptides and adenine are combined with soybean polyphenols to produce derivatives.

The suspension with the branched cyclodextrin is preferably stirred.

Protease is added to this suspension. The protease to be added is preferably 0.001 to 0.2 g per 100 g of the fermentation broth. It is preferable that the protease is suspended in purified water because the reaction proceeds.

This suspension is preferably warmed and stirred to promote the reaction. As heating, 37-44 degreeC is preferable. Moreover, stirring is preferably 10 to 30 times per minute. The time is preferably 1 to 6 hours.

This protease reaction solution is filtered. Efficient filtration is achieved by using filter paper or membrane filter. It is preferable because unreacted components and raw materials can be excluded by filtering to obtain a filtrate.

The obtained reaction product is sterilized by boiling to inactivate the protease.

The obtained reaction product is pulverized by lyophilization and used.

Separating and purifying the target polyphenol derivative from the reaction product is preferable from the viewpoint that the intake can be reduced as a highly pure substance. As a purification method, it is preferable to use a purification operation such as a separation resin.

For example, the desired polyphenol derivative can be obtained by separation with a separation carrier or resin and fractionation. As the separation carrier or resin, porous polysaccharides, silicon oxide compounds, polyacrylamide, polystyrene, polypropylene, styrene-vinylbenzene copolymers, etc., whose surfaces are coated as described later, are used. Those having a particle size of 0.1 to 300 μm are preferred. The finer the particle size, the higher the accuracy of the separation, but the longer the separation time.

For example, a reverse phase carrier or resin whose surface is coated with a hydrophobic compound is used for separation of a highly hydrophobic substance. Those coated with a cationic substance are suitable for the separation of anionically charged substances. Also, those coated with an anionic substance are suitable for separating a cationically charged substance. When a specific antibody is coated, it is used as an affinity carrier or resin for separating only a specific substance.

The affinity carrier or resin is used for specific preparation of an antigen using an antigen-antibody reaction. A partitionable carrier or resin is used for isolation of a substance such as silica gel (manufactured by Merck) if there is a difference in partition coefficient between the substance and the solvent for separation.

Among these, an adsorbent carrier or resin, a dispersible carrier or resin, a molecular sieve carrier or resin, and an ion exchange carrier or resin are preferable from the viewpoint of reducing production costs. Furthermore, the reverse phase carrier or resin and the dispersible carrier or resin are more preferable because the difference in the distribution coefficient with respect to the separation solvent is large.

When an organic solvent is used as the separation solvent, a carrier or resin having resistance to the organic solvent is used. Moreover, the carrier or resin used for pharmaceutical manufacture or food manufacture is preferable.

From these points, Diaion (Mitsubishi Chemical Co., Ltd.) and XAD-2 or XAD-4 (Rohm and Haas) are used as the adsorptive carrier, and Sephadex LH-20 (Amersham Pharmacia) is used as the molecular sieve carrier. Silica gel as the distribution carrier, IRA-410 (Rohm and Haas) as the ion exchange carrier, and DM1020T (Fuji Silysia) as the reverse phase carrier are more preferable.

Of these, Diaion, Sephadex LH-20 and DM1020T are more preferred.

The obtained extract is dissolved in a solvent for swelling the carrier for separation or the resin before separation. The amount is preferably from 1 to 32 times, more preferably from 5 to 19 times the weight of the extract from the viewpoint of separation efficiency. The separation temperature is preferably 4 to 30 ° C., more preferably 10 to 26 ° C. from the viewpoint of the stability of the substance.

As the separation solvent, water or a lower alcohol containing water, a hydrophilic solvent, or a lipophilic solvent is used. As the lower alcohol, methanol, ethanol, propanol and butanol are used, and ethanol used for food is preferable.

When Sephadex LH-20 is used, a lower alcohol is preferable as the separation solvent. When silica gel is used, the separation solvent is preferably chloroform, methanol, acetic acid or a mixture thereof.

When Diaion and DM1020T are used, the separation solvent is preferably a lower alcohol such as methanol or ethanol or a mixed solution of lower alcohol and water.

It is preferable to collect the fraction containing the polyphenol derivative and remove the solvent by drying or vacuum drying to obtain the target polyphenol derivative as a powder or a concentrated liquid because the influence of the solvent can be excluded.

Further, it is preferable to powder this polyphenol derivative for the purpose of preserving.

Hereinafter, the embodiment will be specifically described with reference to examples and test examples. These are merely examples, and conditions can be changed within the range of common sense according to differences in materials, raw materials, and specimens.

Purchased 10 kg of hog-derived placenta from Hokkaido. This was washed with water and then suspended in a pulverizer (Super Free Mill manufactured by Nara Machinery Co., Ltd.) together with purified water to obtain 9 kg of a suspension.

In addition, the pig used what passed the hygiene inspection as quarantined food, and the placenta used frozen and stored.

Furthermore, 10 kg of soybeans from Japan were purchased from the tree of life and used. This was washed and then pulverized by a pulverizer to obtain a pulverized product.

5 kg of the placenta suspension and 2 kg of soybean powder were transferred to a clean stainless steel cylinder, and 20 liters of purified water was added and suspended.

This was sterilized by boiling at 95-97 ° C. for 1 hour. These were transferred to a stirred fermentation tank (FP211) manufactured by Yokogawa Electric Corporation with a capacity of 100 kg, and 20 liters of sterilized purified water was added.

To this, 10 g of powdered Bacillus natto manufactured by Takahashi Yuzo Laboratory Co., Ltd., which is the natto fungus main office, was purchased. This Bacillus natto was suspended in 100 g of sterilized water, powdered soybean powder was added thereto, and the mixture was heated at 37 ° C. for 1 hour and precultured.

The natto fungus solution was added to the agitated fermentation tank and fermented at 40 to 42 ° C. for 42 hours. The state of fermentation was monitored by degradability of soybean powder and quantification of dissolved protein (Burelet method).

After fermentation, the supernatant of the obtained fermentation broth was roughly filtered through a filter cloth to obtain a filtrate.

10 liters of this filtrate was transferred to a clean tank, and 500 g of branched cyclodextrin (Iso Elite) manufactured by Shisui Minato Seika Co., Ltd. was added thereto and stirred sufficiently.

Furthermore, 10 g of protease M “Amano” SD manufactured by Amano Enzyme was added, and the mixture was heated to 37 ° C. and stirred.

Stirring was performed at room temperature for 6 hours using a stirrer. The reaction solution was sterilized by boiling to deactivate the enzyme. The obtained reaction liquid was subjected to suction filtration with Toyo filter paper (No. 2) to obtain a filtrate.

This solution was freeze-dried with a freeze dryer (freeze trap VA-140S manufactured by Taitec Co., Ltd.) to obtain 199 g of the desired powder. This was designated as Sample 1.

70 g of the obtained powder of the specimen 1 was suspended in 200 mL of purified water and subjected to 500 g of Diaion (manufactured by Mitsubishi Chemical) swollen with 5% ethanol. After washing with 700 mL of 5% ethanol, it was further washed with 50% ethanol.

To this, 500 mL of 80% ethanol was added to fractionate the desired polyphenol derivative. The obtained fraction was dried with a vacuum dryer to obtain 21 g of powder. This powder was designated as Sample 2.

Below, the test method regarding the structural analysis of a polyphenol derivative and a result are demonstrated.
(Test Example 1)

The specimen 2 obtained as described above was dissolved in purified water, filtered, and analyzed by high performance liquid chromatography with a mass spectrometer (HPLC, Shimadzu Corporation).

Furthermore, it analyzed with the nuclear magnetic resonance apparatus (NMR, the Bruker make, AC-250). As a result of structural analysis, a derivative in which isoflavone, arginine, and adenine were bound was detected from specimen 2.

The bond was one molecule at a time.

Moreover, the guanidine side chain and amino group of arginine were free.

The confirmation test for keratin production using human skin-derived epidermal cells is described below.
(Test Example 2)

Human skin-derived epidermal cells (HEK cells) were purchased from Toyobo Life Science Division. The epidermal cells were cultured in a dedicated basic medium at 37 ° C. under 5% carbon dioxide gas. Cells in the growth phase were detached with trypsin-containing medium. First, the number of viable cells was counted under a microscope by trypan blue dye exclusion method.

The number of cells was adjusted to 1000 per mL, seeded in 5 mL culture dishes, and further cultured at 37 ° C. under 5% carbon dioxide gas. This was irradiated with ultraviolet rays by an ultraviolet irradiation device (Loctite, output 88 MH) to damage the cells. Irradiation was carried out for 1 hour with the petri dish lid removed.

The epidermal cells were damaged by this ultraviolet irradiation, and recovery from this damage was tested. In addition, this method is a method implemented for evaluation of a test substance in the skin region.

Specimen 2 as a test substance and human EGF (manufactured by Funakoshi) as a test substance were both suspended in physiological saline, diluted and added to a final concentration of 0.1 mg / mL.

Note that physiological saline was used as a solvent control. This was cultured at 37 ° C. for 3 days, and the number of viable cells was counted under a microscope. Further, the cells were dispersed in purified water, and a cell dispersion was obtained using an ultrasonic crusher. The amount of keratin contained in the cell dispersion was quantified by ELISA (Cosmo Bio Inc.).

As a result, the number of epidermal cells increased to 235% by adding Sample 2 with the cell number of the solvent control as 100%. On the other hand, EGF is 140%. Sample 2 was superior.

Regarding the amount of keratin, the value of the solvent control was set to 100%, and the amount of keratin increased to 388% by adding Sample 2. On the other hand, EGF was 211%, and Sample 2 was superior in keratin production.

Since the polyphenol derivative obtained in the present invention exhibits keratin-increasing action and has few side effects, it is used for the treatment and prevention of skin diseases and can improve the national QOL.

Since the method for producing the polyphenol derivative obtained in the present invention can also be used as a food, it contributes to the development of the food industry.

The polyphenol derivative obtained in the present invention is also used as a cosmetic for skin improvement and contributes to the development of the cosmetic industry.

Claims (2)

The polyphenol derivative which exhibits the keratin increase effect | action shown by following formula (1).

A method for producing a polyphenol derivative having a keratin-increasing action comprising a step of adding a branched cyclodextrin to a fermentation broth fermented with natto and adding placenta and soybeans and subjecting to a protease treatment.