JP2016060719A - Pelargonidin derivatives exhibiting melanin decomposition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a pelargonidin derivative exhibiting melanin decomposition.SOLUTION: A pelargonidin derivative exhibiting melanin decomposition consists of pelargonidin, arginine, and glucose. The pelargonidin derivative decomposes melanin by an anti-oxidant action and enzyme reaction, and can remove active oxygen by inducing SOD enzyme. This production method comprises a step of carrying out protease treatment of a fermented solution obtained by adding Monascus to Orchidaceae flowers and rice bran powders and fermenting them. This can be applied in cosmetics, food formulations, and pharmaceuticals. Particularly, if it is utilized in cosmetics, excellent beautiful skin effects including skin whitening, suppleness and luster improvement, and skin elasticity are provided.SELECTED DRAWING: None

Description

The present invention relates to a pelargonidin derivative exhibiting a melanin decomposing action.

Melanin is a brown or black pigment that is produced by an enzyme from proline, a kind of amino acid. Melanin is produced by melanocytes and accumulates in the basal layer of the skin.

The accumulation of melanin in the skin is induced by exposure to sunlight. Sunlight induces the enzyme tyrosinase. In addition, dopaquinone, a product from tyrosine, reacts with cysteine to form melanin.

Although methods and substances for suppressing melanin production have been invented for the purpose of whitening, melanin production has not been suppressed.

For example, there is an invention relating to an inhibitor of tyrosinase activity containing an ethanol-soluble component extracted from white currant as an extraction material or a hydrolyzate thereof as an active ingredient (see, for example, Patent Document 1).

In addition, there is an invention related to a tyrosinase activity inhibitor comprising a quinoveon reductant or a mixture thereof (see, for example, Patent Document 2). However, its range of use is limited and no melanin-reducing action has been shown.

Japanese Patent No. 4886878 Japanese Patent No. 4279523

Inhibition of melanin production by existing plant extracts and chemical substances is mild, and there is a problem that its use in industry is limited. Also, chemically synthesized substances have problems in safety and use is limited. It has been.

Therefore, a pelargonidin derivative having an excellent melanolytic action with weak side effects is provided.

In order to achieve the above object, the invention described in claim 1 relates to a pelargonidin derivative exhibiting a melanolytic action.

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

According to the extract according to claim 1, an excellent melanin decomposing action can be obtained.

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

First, a pelargonidin derivative exhibiting a melanolytic action is a compound having a structure represented by the formula (1).

This pelargonidin derivative consists of one molecule of pelargonidin, three molecules of D-glucose and one molecule of L-arginine.

An ester bond is formed with the hydroxyl group at the 1-position of glucose at the 4-position of the benzene ring located at the 2-position of pelargonine.

The hydroxyl group at the 3-position of pelargonine is ester-bonded to the amino group of guanidino of L-arginine.

Furthermore, the hydroxyl groups at the 5-position and 7-position of pelargonine are both ester-bonded to the hydroxyl group at the 1-position of glucose. By adopting such a structure, the sulfur group derived from cysteine in melanin is cleaved at the molecular level and polymerization of melanin is suppressed.

In addition, the hydroxyl group of glucose relaxes the oxidation reaction of melanin, and the reactivity of melanin is lowered.

Furthermore, the pelargonidin derivatives shown here activate macrophages that phagocytose melanin, enhance the action of phagocytosing and digesting the produced melanin, and reduce melanin.

In addition, pelargonidin derivatives inhibit tyrosinase in melanin producing cells. Its function is attributed to the excellent antioxidant action of pelargonidin derivatives.

In addition, pelargonidin derivatives induce superoxide dismutase (SOD) and activate it to remove active oxygen. In particular, it is preferable that the side chain of arginine activates SOD because it leads to a whitening effect.

Now, pelargonidin as a basic skeleton is a pigment contained in colored flowers of plants such as orchids, grapes, pomegranates, figs, raspberries, blueberries, maquiberries, plums, and carps, and is a kind of polyphenol. It is also a kind of anthocyanidin.

Pelargonidin has a molecular weight of 271.24, is composed of 15 carbons, 11 hydrogens and 5 oxygens, and is 1-benzopyrylium-3,5,7-trihydroxy-2- (4-hydroxyphenyl).

Pelargonidin is preferable in that it has an antioxidative effect and has a function of beautifying skin in addition to melanin decomposing action because it has been reported to protect against ultraviolet rays.

Pelargonidin is preferable because it has abundant dietary experience, does not cause allergies, has not been reported with other side effects, and is highly safe.

D-glucose and L-arginine used in the pelargonidin derivatives shown here are both known substances and have high safety. Moreover, when it takes in excessively, a pelargonidin derivative is decomposed | disassembled by enzymes, such as an esterase in a body, is decomposed | disassembled into pelargonidin, D-glucose, and L-arginine, is metabolized, and excreted, and its safety is high.

Furthermore, it is neutral to the pelargonidin derivative shown here, and there are few side effects on the digestive tract such as the skin, oral cavity and stomach.

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

In the case of the enzyme reaction method, a plant containing pelargonidin or a synthesized pelargonidin is used as a raw material. It is obtained by adding glucose and arginine to this, and further reacting by adding an ester-forming enzyme.

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 is preferably 1 to 35 times the weight of the extract from the viewpoint of separation efficiency, and more preferably 4 to 25 times the amount. 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 pelargonidin derivative acts directly on melanocytes existing in human skin and melanin itself to decompose melanin and exhibits a skin-beautifying action.

It is preferable to add fats and oils to the pelargonidin derivative because the resulting active moiety 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.

As a food preparation, it is used in beauty foods for beauty purposes, foods for beauty purposes, and nourishing tonics for the purpose of maintaining liver cells. Moreover, as a health functional food, it is preferable to use it for a nutrition functional food or a food for specified health.

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

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, skin lotion, etc., and becomes a cosmetic that exhibits a skin beautifying effect.

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

For example, this pelargonine derivative is produced by production comprising a step of subjecting a fermented liquid fermented by adding orchid flower, rice bran powder and Benicowius fungus to protease treatment.

The raw materials are orchid flowers, rice bran powder, benichomycetes and protease.

The orchid here has the scientific name Cornus officinalis.

Orchids are preferred as a raw material for this derivative because they are rich in pelargonine.

Orchids can be from any country such as Japan, China, Taiwan and the United States. In addition, those produced with low pesticides or reduced pesticides are preferred.

Orchids are preferably dried and pulverized, and autoclaving prior to fermentation is preferable because the fermentation can be performed smoothly.

A powder having a particle size of 3 micrometers or less is preferable because it facilitates the fermentation process.

The rice bran powder used as a raw material can be used in rice bran from any origin, such as from Japan, China, the United States, and Russia. However, it is preferable that the powder be produced in Japan because the traceability is reliable and the producer is clear.

Among these, rice bran cultivated organically or without agricultural chemicals is more preferable because it does not contain harmful agricultural chemicals or metals.

In use, KOMENUKA is a free-form mill, super-free mill, sample mill, goblin, super-clean mill, micros, vacuum dryer from Toyo Riko 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.

Furthermore, it is preferable that orchids and rice bran be sterilized after being pulverized by an autoclave or the like because propagation of germs can be prevented.

The fungus used is a filamentous fungus of the scientific name Monasuc purpureus and has long been used in fermented foods such as red sake and tofu in Japan, China and Taiwan.

In addition, bacterial species from Japan such as Okinawa and Kagoshima, and from Southeast Asia in China and Taiwan are used.

This fungus is obtained by fermenting pelargonin consisting of orchids and rice bran.

The amount of each of the above fermentations is preferably 0.2-4 wt. For rice bran powder and 0.0002-0.007 wt. It is preferable to pre-cultivate B. subtilis before fermentation because the initial time of fermentation is shortened and the fermentation time is shortened.

The fermentation is carried out in a clean culture tank, and it is preferable to mix the materials with sterilized tap water.

Moreover, this fermentation is heated at 36-47 degreeC, and fermentation is performed for 15 days from 1 day. The process of fermentation is controlled by quantifying the target pelargonine derivative by HPLC or TLC, and confirming the growth of cells.

Moreover, since the product is easy to isolate | separate, it is preferable to ultrasonically treat the obtained fermented material. Moreover, it is preferable to concentrate by freeze drying or the like because the following steps can be performed in a short time.

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.

Orchids grown with reduced pesticides in Okinawa were used. After collecting the flowers, they were washed with tap water, dried in the sun, and pulverized with a pulverizer (Super Free Mill manufactured by Nara Machinery Co., Ltd.) to obtain 1.2 kg of dry powder pulverized orchids.

Rice-derived rice bran from Mie Prefecture was supplied to a mixer (Cuisinate) to obtain 1.2 kg of rice bran pulverized material. The pulverized orchid flower and rice bran were put in an autoclave and sterilized at 121 ° C. for 20 minutes.

These were put into a clean fermentation tank (sterilized round 40-liter tank for fermentation), and 11 kg of sterilized tap water was added and stirred.

Separately from this, 11 g of powdered B. cerevisiae (manufactured by Natto Honpo) was subjected to a small fermentation tank, and a sterilized rice bran powder and a pre-cultured culture solution were prepared.

The pre-cultured B. niger solution was added to the fermentation tank containing the dried orchids and rice bran, and after stirring, the mixture was heated in a temperature range of 39 to 42 ° C. for fermentation.

In the fermentation process, the fermentation liquor was sampled while bubbling and stirring by aeration.

To 1 kg of the obtained fermented product, 11 g of Amano Protease N was added and heated at 40 ° C. for 3 hours.

This treated product was heated, and ethanol was added to obtain 230 g of the desired pelargonine derivative. This was designated as Sample 1.

(Test Example 1)
The specimen 1 obtained as described above was dissolved in ethanol 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, the specimen was composed of one molecule of pelargonidin, three molecules of D-glucose and one molecule of L-arginine.

It was ester-bonded with the hydroxyl group at the 1-position of glucose at the 4-position of the benzene ring located at the 2-position of pelargonine.

The hydroxyl group at the 3-position of pelargonine was ester-linked with the amino group of guanidino of L-arginine.

Furthermore, the hydroxyl groups at the 5th and 7th positions of pelargonine were both ester-bonded to the hydroxyl group at the 1st position of glucose.

The confirmation test using human skin-derived melanin producing cells is described below.
(Test Example 2)

Human-derived melanocytes (melanocells) purchased from Kurabo Industries Co., Ltd. were used. As a culture solution, 1000 cells cultured using 5% fetal calf serum-containing MEM medium (manufactured by Sigma) were seeded in a 35 mm culture dish, and cultured at 37 ° C. under 5% carbon dioxide gas.

To this, the specimen 1 obtained in Example 1 and vitamin C as a control were added at a final concentration of 0.1 mg / ml. This was cultured for 48 hours.

After detaching the cells, the number of cells was counted. In addition, the petri dish calculated the average value using five sheets.

As a result, the addition of 0.1 mg / ml of Specimen 1 reduced the number of melanin producing cells to 33% as an average value compared to the solvent control group. On the other hand, when vitamin C was added, the number of cells decreased to 64%.

The obtained cells were collected and sonicated to collect a solution containing melanin. The amount of melanin in this solution was quantified by HPLC (manufactured by Shimadzu Corporation).

As a result, the amount of melanin due to the addition of specimen 1 was 11% compared to the value of the solvent control. On the other hand, the addition of vitamin C was 57%. As a result, specimen 2 was found to have a markedly lower melanin inhibitory effect than vitamin C.

The melanin degradation test is described below.
(Test Example 2)

Melanin obtained from human-derived melanin-producing cells as described above was prepared at a concentration of 0.1 mg / mL, 10% of the sample 1 solution was added thereto, and the mixture was stirred at 37 ° C. for 3 hours. Thereafter, the amount of melanin was quantified.

At the same time, a control group to which the same amount of solvent was added was used. As a result, the addition of specimen 1 resulted in a melanin amount of 15% compared to the solvent control, indicating a decrease in melanin.

The pelargonidin derivative obtained in the present invention exhibits a melanin decomposing action and has few side effects. Therefore, it improves the QOL of the people and contributes to the development of the beauty industry.

Claims (1)

A pelargonidin derivative having a melanin decomposing action having a structure represented by the formula (1).