JP2017141205A - Phenol derivative having plasma fluidity increase action and method for producing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a phenol derivative having a plasma fluidity increase action, and a method for producing the same.SOLUTION: Provided is a phenol derivative composed of hydroxy sesamol, erythritol and tyrosine. Also provided is a production method comprising a process where a fermentation liquid obtained by adding sesame seed and soybean powder with Bacillus natto made of Natto Honpo and fermenting the same is fermented with Monascus purpureus. The obtained phenol derivative has an excellent plasma fluidity increase effect, generates a hydrogen gas, and is applied to cosmetics, food preparations and medicines. In the cosmetics, the plasma fluidity of skin cells is increased, and in the food preparations, the protective action of nerve cells is exhibited.SELECTED DRAWING: None

Description

The present invention relates to a phenol derivative exhibiting a plasma fluidity increasing action and a method for producing the same.

Cell function and normal growth involve normal gene status, normal energy transfer and intracellular signal transduction, normal substance transfer and cytoskeleton maintenance, etc. Yes. In particular, plasma fluidity is involved in maintaining these cellular functions.

Plasma fluidity mainly involves tubulin proteins, microtubules, intermediate filaments, actin, and myosin that control the cytoskeleton. In addition to this, ions such as ATP production, mitochondrial function, endoplasmic reticulum, calcium ions, etc. In addition to fluidity, it is closely related to the total function including moisture.

The discovery of protoplasmic fluidity itself is in the 20th century, and the research results are old, but there are many unclear points about their functionality. For this reason, research and industry have not made progress on the relationship between cell function and protoplasmic fluidity. Moreover, it has not been able to identify a protoplast fluid activator or a substance that acts to suppress it.

Among these, for example, although there is a composition containing microvesicles derived from cell protoplasts as an invention related to cell protoplasts, there is no explanation about protoplasm fluidity in this invention (for example, patent document) 1).

Further, as an invention relating to a drug discovery screening method and apparatus, a method of distinguishing protoplasmic flow from Brownian movement and determining the activity of a cell based on the protoplasmic flow has been invented. It is not an invention relating to this, and no substance relating to its activation is described (for example, see Patent Document 2).

Special table 2013-534830 Japanese Patent No. 4507060

The effect of increasing protoplasmic fluidity by existing substances is mild, and there is a problem that its use in industry is limited, and there is a problem in safety with chemically synthesized substances, and its use is limited .

Therefore, a natural product exhibiting an excellent action for increasing protoplasmic fluidity with weak side effects and a production method for efficiently producing the same are desired.

In order to achieve the above object, the invention described in claim 1 relates to a phenol derivative having an action of increasing protoplasmic fluidity 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 phenol derivative having a protoplasmic fluidity increasing action.

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

The phenol derivative according to claim 1 is excellent in the plasma fluidity increasing action.

According to the manufacturing method of Claim 2, it is possible to manufacture the phenol derivative which exhibits a plasma fluidity increase effect efficiently.

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

The phenol derivative exhibiting a protoplasmic fluidity increasing action has a structure represented by the following formula (1).

As shown in the above formula (1), it consists of one molecule of hydroxysesamol, one molecule of erythritol, and one molecule of tyrosine. These bonds are all natural and are linked through ester bonds and ether bonds.

This phenol derivative can be obtained by chemical synthesis using hydroxy sesamol, erythritol, tyrosine and the like as raw materials by chemical synthesis. However, the chemical synthesis is limited in industrial use due to the significant loss of raw materials. Chemical synthesis is preferable for obtaining a standard product or a small amount of a sample of this phenol derivative.

It is preferable to analyze the structure of this phenol derivative from the viewpoint that an active ingredient can be specified. Further, it is preferable because it can be used as an index of the content of an active ingredient when it is used and sold in products and preparations.

As an example of the structural analysis of this phenol derivative, the peak positions are 3.079, 3.432, 3.781, 3.814, 5.727, 6 by 400 MHz H-NMR in deuterated chloroform, for example. 0.021, 6.138, 6.302, 6.549, 6.952, 7.137, 7.376, 7.462, 7.484, 7.8446 and 7.901 ppm.

Furthermore, this phenol derivative is analyzed by high performance liquid chromatography or a mass spectrometer, and its structure is identified.

Hydroxysesamol which is a constituent component is a hydroxy form of sesamol, is one of the constituent components of sesamin contained in plants such as sesame, and is a phenol used in plants and animals. Its chemical formula is C7H6O4.

This hydroxy sesamol has a hydroxyl group at the 1-position and 3-position of the complex benzene ring, and has a strong antioxidant property, antibacterial property and enzyme activation action by two phenolic hydroxyl groups. In this phenol derivative, both hydroxyl groups of hydroxysesamol are bound, but they bind to tubulin, actin, and myosin related to protoplasmic fluidity, and act to increase the actin myosin binding and ATP utilization rate. Has an effect of increasing plasma fluidity.

In this phenol derivative, one of the hydroxyl groups of hydroxysesamol is ester-bonded to the carboxyl group of tyrosine. The other hydroxyl group is ether-bonded to the hydroxyl group of erythritol.

Since hydroxysesamol itself is less toxic and exhibits both water-soluble and fat-soluble properties as compared to sesamol, it is preferable to exhibit both properties of cell membrane permeation and protoplast lysis.

Moreover, it is preferable from the point which suppresses the aging and oxidation of a cell that a hydroxy sesamol part can pass through a mitochondrial membrane, and can eliminate the active oxygen produced in ATP production. This antioxidant power results in increased cell fluidity and cell stabilization.

In addition, yeasts and useful microorganisms used for fermentation increase their protoplasmic fluidity and have the effect of producing useful substances and promoting fermentation. This action of accelerating fermentation can be used for fermentation processes with low reactivity and for substance production by fermentation. That is, it is preferable that production of a rare substance can be promoted by promoting fermentation with poor reactivity.

In addition, erythritol, which is a constituent component, is a kind of sugar alcohol having the chemical formula C4H10O4. It is a natural low-calorie sweet substance contained in fruits such as melon, vegetables and natto, and fermented products such as sake and soy sauce.

This erythritol is a natural product and preferably has high safety. Furthermore, since the hydroxyl group of erythritol exhibits a reducing action, it is preferable from the viewpoint of component stability that it contributes to the removal of active oxygen and the stability of the substance.

In this phenol derivative, the phenolic hydroxyl group of hydroxysesamol and the hydroxyl group of erythritol are ether-bonded.

The constituent tyrosine has the molecular formula C9H11NO3, L-type, and natural type. Tyrosine is a form in which one phenolic hydroxyl group is bonded to phenylalanine, and is classified as a non-essential amino acid.

Since tyrosine is induced into neurotransmitters dopa, dopamine, noradrenaline and adrenaline, it is involved in the action on nerve cells. That is, the tyrosine moiety is preferable from the viewpoint of protecting peripheral nerves and brain function.

The tyrosine of this phenol derivative is necessary for exerting an antioxidant action by the phenolic hydroxyl group and increasing protoplasmic fluidity.

Since this phenol derivative exhibits both fat-soluble and water-soluble properties, it passes through the cell membrane and cell wall and is absorbed into the cell. This phenol derivative binds to tight junction proteins such as tubulin, actin and myosin, and occludin in the cytoplasm, and to the plasma membrane-lined protein, and increases plasma fluidity.

This phenol derivative increases the plasma fluidity by promoting the contraction and relaxation of the cell skeleton by promoting the binding of actin and myosin. That is, the ATP reaction site of actin and myosin is activated at the binding site of tyrosine and hydroxysesamol of this phenol derivative.

Furthermore, the structure of the phenol derivatives hydroxysesamol and erythritol increases the sliding and loosening of actin and myosin, increasing the relaxation and contraction of actin and myosin and increasing the plasma fluidity.

Furthermore, the active center of the ATP-producing enzyme is activated by the hydroxyl group of erythritol and the amino group of tyrosine to increase glycolytic ATP production, thereby increasing the amount of local ATP that is the driving force for the increase in plasma fluidity.

Increased protoplasmic fluidity causes intracellular ATP, vitamins, minerals, water, proteins, lipids, carbohydrates and other nutrients, enzymes and growth factors, and cyclic AMP and cyclic components of intracellular signal transduction systems Move GMP etc. This movement of nutrients and growth factors increases cell reactivity and activates cell functions.

Increased protoplasmic fluidity also increases cell membrane fluidity and normalizes growth factor receptor migration and reactivity. Furthermore, this phenol derivative is preferable because it contributes to normalization and maintenance of cell function by activating the movement of genes such as transcription factors that move through the cell nuclear pore, enzymes and mRNA and contributing to appropriate transmission of genetic information. .

Since protoplasmic fluidity is abnormal in pathological cells and cancer cells, this increase in protoplasmic fluidity caused by this phenol derivative will normalize the cells and improve diseases such as cancer at the cellular level. Phenol derivatives are preferred.

It is also suitable for functions related to the excretion of toxins and waste products in epithelial cells and liver cells of kidney cells, and by increasing the plasma fluidity, waste products such as waste products and oxidants can be efficiently discharged outside the cell. Excrete. This phenol derivative is preferred because it has a detox effect that purifies cells by excretion.

In addition, it promotes the movement and degradation of phagocytic tissues, foreign bodies, bacteria, and viruses by increasing the protoplasmic fluidity of phagocytic cells such as macrophages, monocytes, Langerhans cells, mesangial cells, and Kupffer cells. This is because this phenol derivative is preferable from the viewpoint of an increase in immune function and suppression of allergic reaction.

In neuronal cells, axonal transport is related to protoplasmic fluidity, so that growth factors such as NGF are transported, and that neurotransmission is active is preferable because it normalizes the function of neuronal cells. This phenol derivative is also preferable from the viewpoint of activation of neurotransmission in cranial nerve cells such as peripheral neuropathy and dementia and excretion of amyloid accumulated in the brain.

In addition, since this phenol derivative contains abundant hydroxyl groups, it has an excellent hydrogen gas generating action and antioxidant power, and it suppresses the production of melanin to bring about a skin whitening effect. It is preferable because it increases. That is, the phenol derivative powder reacts with a water-soluble solvent to generate hydrogen gas. The hydrogen gas concentration of this aqueous solution is 1.6 ppm.

In myocardial infarction, this phenol derivative exerts an effect on prevention and treatment of myocardial infarction by activating mass transfer of cardiomyocytes even in the coronary artery infarction or ischemic state. In particular, this phenol derivative exerts a relaxing action in vascular smooth muscle at the infarcted site to improve blood flow and blood pressure. Since hydrogen gas removes active oxygen generated during ischemia reperfusion, it is preferable to generate hydrogen gas.

Moreover, it is preferable that this phenol derivative raises the activity of the muscle at the time of exercise | movement by producing ATP via a glycolysis system in an apnea state. In particular, when an athlete wants to strengthen muscles, it is preferable because it increases the intracellular movement of glucose, vitamins, minerals, and oxygen in muscle cells, transports them to mitochondria, and excretes waste products, carbon dioxide, and active oxygen. Moreover, since active oxygen is generated in mitochondria, it is preferable to eliminate it with hydrogen gas.

This phenol derivative is degraded in vivo by kidney and liver esterases and excreted in urine. It is decomposed into highly safe hydroxy sesamol, erythritol and tyrosine, which are constituents. Therefore, this phenol derivative is not accumulated in the body, is decomposed by an in vivo enzyme, and the decomposed product is also a natural product, so it is highly safe.

This phenol derivative easily penetrates into the fat cell membrane, moves the neutral fat of fat cells, enhances this degradation, and degrades carbohydrates. Since carbohydrates are consumed, it is preferable for diabetes prevention and diet countermeasures.

Furthermore, this phenol derivative also suppresses inflammation of skin epithelial cells and suppresses the formation of wrinkles. In addition, it stabilizes the keratinocytes to maintain the skin keratin barrier function and suppress the entry of foreign substances, irritants and bacteria. This function can be used as a cosmetic.

Since this phenol derivative activates cells such as macrophages and lymphocytes by increasing plasma fluidity, it can be used as an immunostimulant or antiallergic agent. In addition, this phenol derivative works on nerve cells, and is suitable as a therapeutic agent for dementia, Alzheimer's disease, and Parkinsonism by increasing cell membrane potential and nerve conduction.

In addition, this phenol derivative is a substance in which all decomposed components are present in nature, and its safety has been confirmed because of its abundant experience in food use and cosmetics. Furthermore, this phenol derivative restores and promotes the function of cells by conjunctivitis, cataracts, glaucoma, and retinal detachment by activating the cytoskeleton of eye corneal cells, lens cells, and retinal cells, and is used to restore vision. The

This phenol derivative also exists in nature and is found in trace amounts in sesame seeds. This phenol derivative can be extracted from the above plants by purification. However, refining requires a large amount of raw materials and uses organic solvents, which limits the industrial use.

This phenol derivative preferably increases sesame seeds by fermentation or the like. As a fermentation method, it is obtained by mixing with soybeans and fermenting with Bacillus natto or Bacillus subtilis. This method is preferable because it has dietary experience and the production amount of phenol derivatives is large.

When the obtained phenol derivative is used as a pharmaceutical material, it is preferable to purify the target phenol derivative because the purity of the target phenol derivative is increased and impurities can be removed.

Used as pharmaceuticals as parenterals such as injections, oral preparations or coatings, and quasi-drugs used in tablets, capsules, drinks, soaps, coatings, gels, toothpastes, 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 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.

Dietary supplements that increase cell fluidity and improve immunity by increasing plasma fluidity as food preparations, detox and nourishing tonic foods, beauty supplements that maintain whitening and skin health, nerves, liver and kidneys It is used for health foods that improve functions, health foods for the purpose of dieting to strengthen muscles and decompose fats, and beauty foods. 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 pet supplement for the purpose of strengthening detox, whole body epithelial tissues, muscles, and bone cells.

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, cosmetic liquid, lotion and the like.

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

It is preferable to prevent wrinkles and sagging by promoting the proliferation of skin cells by promoting the protoplasmic fluidity even in a hypoxic state, and promoting the production of keratin and collagen. Furthermore, the whitening effect by suppressing the production of melanin by the antioxidant action is exhibited.

In addition, this phenol derivative exhibits antibacterial and antioxidative effects due to the phenolic hydroxyl group, and can be used as a dentifrice, mouthwash or toothpaste for the purpose of suppressing inflammation and proliferation of gingival cells.

Furthermore, it can be utilized as a fermentation activator that activates fermentation by increasing the protoplasmic fluidity of useful microorganisms and yeasts. This fermentation activator can activate a fermentation stage with low reactivity to produce useful ingredients and foods. For example, production efficiency can be increased at low cost by making the fermentation process more efficient in the fermentation process of pharmaceutical raw materials.

Moreover, it can utilize as a plant active agent which makes the growth of a plant active by increasing the protoplast fluidity | liquidity of a plant cell. This plant activator can be used for the growth of rare orchids and flowers and activates the cultivation of fruits, vegetables and cereals. It can also be used for cultivation of vegetables and fruits in plant factories, thus increasing cultivation efficiency.

Next, a phenol derivative that exhibits a protoplasmic fluidity-increasing effect consisting of a step of fermenting fermented liquor with fermented sesame seeds, soybean powder and natto Honpo natto bacteria fermented with Benikouji fungus A manufacturing method will be described.

The phenol derivative here is a substance composed of one molecule of hydroxy sesamol, one molecule of erythritol, and one molecule of tyrosine, and these bonds are all natural types, and the bonding mode is an ester bond and an ether bond. This phenol derivative penetrates the intracellular cytoplasm and increases protoplasmic fluidity, thereby suppressing detox, immunity, ischemia, cerebral infarction, myocardial infarction, inflammation and cancer growth.

The phenol derivatives hydroxy sesamol, erythritol, and tyrosine are preferable because they exist in nature, have abundant food experience, and are recognized as safe.

This derivative also works on skin, nerves, bones, muscles, liver, kidneys, etc., and penetrates into cells to increase protoplasmic fluidity, thereby proliferating cells and improving blood flow.

This production method comprises a step of fermenting a fermented liquid fermented by adding sesame seeds, soybean powder and natto honto natto bacteria with Benikouji bacterium from red rice honpo.

The raw materials are sesame seeds, soybean powder, natto bacterium from Natto Honpo, and Benikouji fungus from Kurisu Honpo.

Sesame here, that is, sesame seeds is the annual name Sesamum indicum, the sesame genus Sesame, and its seeds are used for food and have abundant food experience. Although it is classified according to the color of the seed coat, such as white sesame, black sesame and yellow sesame, black sesame seeds are preferred because they contain a large amount of tannin.

In addition, sesame seeds are a traditional Chinese medicine that uses sesame seeds, and is used for the purpose of reducing weak heat. Since sesame seeds contain tannin, polyphenols, organic acids, minerals, pigments, and polyphenols, they are preferable as raw materials for producing this phenol derivative.

Sesame seeds may originate from any country such as Japan, China, Taiwan, and the United States. In particular, those produced in Japan with low pesticides or reduced pesticides are preferred. For example, black sesame seeds cultivated without pesticides by Mitsubachi Ichibantei in Kagawa Prefecture are preferred because of their high quality.

Sesame seeds are preferably dried and pulverized, and sterilization by autoclave prior to fermentation is preferable because 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 soybean powder used as a raw material can be used in soybeans of any origin such as Japanese, Chinese, American, and Russian, but is preferably Japanese because of its reliable traceability and clear producers.

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.

Further, it is preferable that sesame seeds and soybeans are sterilized after being pulverized and then sterilized by an autoclave or the like because propagation of germs can be prevented.

The natto bacterium produced by Natto Honpo is the scientific name Bacillus subtilis, which is widely used in the production of natto in Japan. Bacteria species from Japan such as Okinawa and Kagoshima, and from Southeast Asia such as China and Taiwan are used. The Bacillus natto used is manufactured by Natto Honpo and exhibits high fermentability.

This Bacillus natto promotes the binding reaction of hydroxysesamol, erythritol, and tyrosine, which consist of sesame seeds and soybeans.

The amount of each of the above-mentioned fermentations is preferably 0.04 to 4 wt. For soybean powder and 0.002 to 0.05 wt. It is preferable to pre-cultivate natto bacteria before being fermented, 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 41-44 degreeC, and fermentation is performed for 1 to 7 days. It is preferable to carry out the fermentation process control by quantifying the target phenol derivative by HPLC or TLC and confirming the growth of the bacterial cells.

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

Hydroxysesamol, erythritol, and tyrosine are bound by this fermentation process, but since the bond is unstable, the next fermentation with Benikouji fungus made by Kurisu Honpo is performed to stabilize the bond.

The Benikouji fungus made by Benito Honpo to be 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. Benikouji fungus made by Kurisu Honpo has excellent fermentation efficiency.

As for each addition amount regarding the said fermentation, 0.0003-0.01 weight is preferable with respect to 1 weight of said fermented products. It is preferable to pre-cultivate Benikouji fungus made by Kurisu Honpo before fermentation because it shortens the initial time of fermentation and shortens the fermentation time.

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 to 39-42 degreeC, and fermentation is performed for 1 to 7 days. This fermentation process stabilizes the structure of the phenol derivative by the reducing action of Aspergillus niger.

It is preferable that the fermented product is extracted with water-containing ethanol because the product can be efficiently recovered, the bacteria can be sterilized, and the next step can be easily performed. 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.

Separating and purifying the target phenol derivative from the reduction reaction product is preferable because 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 phenol 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 thereof is preferably 2 to 40 times the weight of the extract from the viewpoint of separation efficiency, and more preferably 4 to 20 times the amount. The separation temperature is preferably 10 to 39 ° C., more preferably 12 to 37 ° 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 a fraction containing a phenol derivative and remove the solvent by drying or vacuum drying to obtain the target phenol derivative as a powder or a concentrated liquid because the influence of the solvent can be excluded.

Moreover, it is preferable to carry out the final extraction with fats and oils used for edible oils and cosmetics because the obtained phenol derivative is stably maintained. For example, extraction with soybean oil, rice bran oil, grape seed oil, olive oil or jojoba oil is preferred.

Further, it is preferable to powder this phenol 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.

Black sesame seeds grown in pesticide-free in Kagawa Prefecture were purchased from Michibachi Ichibatei and used. The seeds 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 1.1 kg of a pulverized dry powder of sesame seeds.

Hokkaido soybean was used in a mixer (manufactured by Kuisinart) to obtain 1.1 kg of soybean ground material. The sesame seeds and soybeans were crushed 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 5 kg of sterilized tap water was added and stirred.

Separately, 11 g of powdered natto bacteria manufactured by Natto Honpo was used in a small fermentation tank, and a sterilized soybean powder and a precultured preparation liquid were prepared.

It added to the fermentation tank which put the fermented preparation liquid of the said Bacillus natto, the dried powder of the sesame seed, and the soybeans which were the said pre-culture, and after stirring, it heated and fermented in the temperature range of 40-42 degreeC.

In the fermentation process, the fermentation liquor was sampled and fermented for 3 days while bubbling and stirring by aeration. After completion of fermentation, the fermented product was taken out from the fermentation tank and sterilized by boiling. This fermented product was filtered through a filter cloth to obtain 1.1 kg of a fermented solution of Bacillus natto. To 1 kg of this fermented liquid, 10 g of Benikouji fungus made by Kurisu Honpo was added and fermented at 41 ° C. for 6 days.

Ethanol was added to the fermented product and sterilized by boiling. This was filtered, and the filtrate was freeze-dried to obtain 0.56 kg of the desired phenol derivative. This was designated as Sample 1. In addition, when 0.1 g of Sample 1 was dissolved in 10 mL of purified water, it was measured by gas chromatography (Shimadzu Corporation) that 1.6 ppm of hydrogen gas was generated.

Furthermore, a purified product was obtained for the purpose of structural analysis and experiment. That is, 1 L of 5% ethanol-containing purified water was added to 200 g of the above-described phenol derivative of Sample 1, and 500 g of Diaion (manufactured by Mitsubishi Chemical) was suspended in a 5% ethanol solution and filled into a glass column.

3 L of 5% ethanol solution was added thereto for cleaning, and 2 L of 70% ethanol solution was added to elute the desired phenol derivative, followed by concentration and purification. The purified phenol derivative was distilled under reduced pressure to remove the ethanol portion to obtain an aqueous solution. This was obtained 22 g of a purified phenol derivative, which was designated as Sample 2. Moreover, when 0.1 g of this specimen 1 was dissolved in 5 mL of purified water, it was confirmed by gas chromatography (Shimadzu Corporation) that 1.6 ppm of hydrogen gas was generated in the aqueous solution.

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

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

As a result of analyzing this with a nuclear magnetic resonance apparatus (400 MHz, H-NMR, manufactured by Bruker), a phenol derivative consisting of one molecule each of hydroxysesamol, erythritol and tyrosine was detected from specimen 2.

That is, 3.079, 3.432, 3.781, 3.814, 5.727, 6.021, 6.138, 6.302, 6.549, 6.549 in deuterated chloroform of H-NMR. Signals were observed at 952, 7.137, 7.376, 7.462, 7.484, 7.846, and 7.901 ppm.

The above analysis results were found to exhibit the same structure as a chemically synthesized standard product. That is, it was confirmed from Sample 2 that the target phenol derivative was obtained by ester-linking one molecule of hydroxysesamol, one molecule of erythritol, and one molecule of tyrosine.

The confirmation test using human skin epithelial cells is described below.
(Test Example 2)

Human-derived skin epithelial cells purchased from Kurabo Industries Co., Ltd. were used. 1000 cells cultured using 5% fetal calf serum-containing MEM medium (manufactured by Sigma) as a culture solution were seeded in a 35 mm culture dish (manufactured by FALCON), and cultured at 37 ° C. under 5% carbon dioxide gas. This was irradiated with ultraviolet rays by an ultraviolet irradiation device (manufactured by Eye Graphics Co., Ltd.). Further, EGF (Funakoshi Co., Ltd., human type) was added at a final concentration of 0.1 mg / ml as Sample 1 and Sample 2 and the positive control. This was cultured for 48 hours and tested.

The protoplasmic fluidity was observed by analyzing the moving images with a high-sensitivity differential interference microscope and video system. That is, while the cells treated with the specimen were placed in a culture dish, a cell image was taken with a high-sensitivity camera at a magnification of 100 using a Hamamatsu Photonics (ORCA-Flash 4.0V2) and a Nikon microscope.

The intracellular protoplasmic fluidity of the cells was observed for 1 minute and edited into a video. The image was analyzed at low speed, and the protoplasmic fluidity was counted from the distance and frequency of movement of intracellular substances. In addition, the petri dish calculated the average value using five sheets. It compared with the solvent control group which added the solvent.

As a result, the addition of 0.1 mg / ml of Specimen 1 increased the number of skin epithelial cells to 143% as an average value compared to the solvent control group. In Sample 2, it increased to 211%. On the other hand, EGF increased by 140%, and Sample 1 and Sample 2 were superior.

The protoplasmic fluidity increased to 166% by Specimen 1 compared to the solvent control group. In addition, the addition of Specimen 2 showed an increase of 334% of the solvent control. In EGF, it became 105%, and the plasma fluidity increasing action by specimen 1 and specimen 2 was remarkable.

Since the protoplasmic fluidity increasing action is also an indicator of cell activity, it was confirmed by the specimen 1 and the specimen 2 that the glycolytic activity was increased by the treatment of the specimen 1 and the specimen 2.

The amount of intracellular keratin was increased to 155% by Sample 1 compared to the solvent control group. Moreover, it became 199% of the solvent control by the addition of the specimen 2. EGF was 151%, and Sample 1 and Sample 2 were superior in increasing keratin production. It was confirmed that 1.6 ppm of hydrogen gas was generated in the culture medium.

The confirmation test using a human nerve cell injury model is described below.
(Test Example 3)

Human neurons (Human Neurons (HN)) purchased from Cosmo Bio were used. As a culture solution, 1000 cells cultured using a dedicated culture solution (neural cell growth medium) were seeded in a 35 mm culture dish and cultured at 37 ° C. in 5% carbon dioxide gas. To this, 1% aqueous acrylamide solution was added to weaken the nerve cells.

To this, Specimen 1 and Specimen 2 obtained in Example 1 above and NGF (Funakoshi Co., Ltd., human type) as a positive control were added at a final concentration of 0.1 mg / ml. This was cultured for 48 hours.

After culturing, the protoplasmic fluidity was measured by microscopic observation as described above in a state where the nerve cells were placed in a culture dish. In addition, the number of cells was counted microscopically. In addition, the petri dish calculated the average value using five sheets. It compared with the solvent control group which added the solvent.

As a result, the addition of 0.1 mg / ml of Specimen 1 increased the number of neurons to 133% as an average value compared to the solvent control group. In Sample 2, it increased to 193%. On the other hand, NGF increased by 130%, and Sample 1 and Sample 2 were superior.

The plasma fluidity of nerve cells was increased to 169% by Sample 1 compared to the solvent control group. The addition of Specimen 2 increased to 266% of the solvent control. NGF was 153%, and Sample 1 and Sample 2 were superior to NGF in increasing plasma fluidity. Furthermore, it was confirmed that 1.6 ppm of hydrogen gas was generated in this culture solution.

The phenol derivative obtained in the present invention increases the protoplasmic fluidity of cells and generates hydrogen gas, thereby increasing the detox function, reducing cancer, inflammatory diseases and health disorders such as skin, liver and nerves, and Because there are few side effects, it can improve the QOL of the people, increase the healthy working population, and reduce medical expenses.

Since the phenol derivative obtained in the present invention has an action of improving skin inflammation, it contributes to the improvement of the skin suffering from skin troubles such as wrinkles and sagging as a cosmetic, and contributes to the development of the cosmetic industry.

The phenol derivative obtained in the present invention can be used as a food because it is produced from plant-derived raw materials by fermentation, and contributes to the development of the food and fermentation industries.

Claims (2)

A phenol derivative exhibiting a plasma fluidity increasing action represented by the following formula (1).

A method for producing a phenol derivative, which comprises a step of fermenting a fermented liquid obtained by adding sesame seeds, soybean powder and natto natto bacteria and fermenting the fermented liquor with Benikouji bacteria manufactured by Kurisu Honpo.