TW201501727A - A revitalizing agent and a collagen generating promoting agent having cellobiose ester as the active component - Google Patents

Abstract

The present invention provides a revitalizing agent that provides excellent anti-aging effects and vitalizing effects to cells, as well as safety from the prolong use of the revitalizing agent and the collagen generating promoting agent. The present invention also provides cosmetics, quasi-drugs, pharmaceutical, food, and beverage products having the revitalizing agent and the collagen generating promoting agent as active ingredient. The cell-revitalizing agent of the instant disclosure is characterized by including cellobiose esters, which is a sugar ester biosurfactant.

Description

Activator and collagen egg with cellobiose ester as active ingredient White production accelerator

The present invention relates to an activator using a cellobiose ester as an active ingredient, and more particularly to a cosmetic, quasi-drug, pharmaceutical, food or drink containing cellobiose ester, which is made by activating various cells It can improve the metabolism, regeneration and repair functions of cells, and is effective for preventing aging or hair growth and preventing hair loss. Furthermore, the present invention relates to a collagen production promoter, and relates to a cosmetic, quasi-drug, pharmaceutical, food or drink containing a bio-surfactant, which is resistant to aging by producing collagen. Adjusts skin texture and prevents or inhibits skin wrinkles.

Various diseases and the like are associated with a decrease in the rate of division of all cells and a decrease in cell function. For example, the dermis and epidermis of the skin are composed of epidermal cells, fibroblasts, and extracellular matrices such as elastin, collagen, and hyaluronic acid supporting the extracellular skin structures. In particular, the synthesis or decomposition of collagen and hyaluronic acid is mainly controlled by fibroblasts. In young skin, the interaction of these skin tissues is maintained constant, thereby ensuring skin properties such as softness, and the appearance of the skin is also glossy, firm, transparent, and maintained in a moisturized state. However, due to UV rays, dryness, Pressure, etc., especially causes a decrease in the function of extracellular matrix and fibroblasts, and as a result, skin properties such as skin softness are lowered, and the skin loses luster, firmness, and transparency, and causes symptoms such as roughness, wrinkles, and dullness. It is also known that as skin properties decline, the function of fibroblasts will decline together, and the metabolic rate of collagen and hyaluronic acid will decrease.

The industry is constantly exploring cell-level activators and anti-aging agents. For example, as an activator derived from an animal system, a connective tissue hydrolyzate, a water-soluble protein derived from the thymus or the spleen, a bovine placenta extract, and the like are known. As an activator from plant lines, sesame, yam, pepper, angelica, houttuynia, maiden, almond, dandelion, western elder, chuanxiong, Swertia japonica, mulberry white are known. , peach kernels, carrots, hops, wood only, coix seed, ginger turmeric, and the extract of the genus Chrysomaphus genus. Some of these are used as quenching agents and anti-aging agents in quasi-drugs or cosmetics. However, since individual differences are very large, the effect is difficult to be sufficient, and the effect of cell activation or anti-aging effect is exerted. Not satisfactory.

A sugar ester-based amphiphilic substance which has both a hydrophilic property derived from a sugar property and a lipophilic property derived from a lipid, and has a function as a surfactant. There are various amphiphilic substances in living things, and they participate in the exchange of substances, energy and information at various interfaces, which play a great role in the formation of biological order.

In recent years, a cellobiose ester (CL) derived from a bio-surfactant (biosurfactant) of a sugar ester has been reported to exhibit antifungal activity (Non-Patent Document 1). Cellobiose ester It is produced by Ustilago maydis or Cryptococcus humicola (Non-Patent Document 2). Further, it is known that Pseudozyma flocculosa can also produce one of the hairy yeasts of the CL (Flocculosin) (Non-Patent Document 3). Further, it is known that Ustilago esculenta can also produce CL in a large amount (Patent Document 1). Since these CLs produced by microorganisms all exhibit excellent antifungal activity, they are expected to be put to practical use as antifungal drugs.

The industrial use of cellobiose esters in various fields such as detergents and cosmetics is progressing, and it is used as a liposome-forming agent (see Patent Document 2), an emulsifier, a solvent (see Patent Document 3), and a carrier for protein separation. (refer to Patent Document 4), low molecular organic organogel (see Patent Document 5), and the like.

Cellobiose ester is expected to be practical as a raw material for cosmetics or pharmaceuticals because it has high biodegradability, low toxicity, and mild environment. However, the effect of cellobiose esters on cells has not been found at present and there is no report of its effect or efficacy.

[Previous Technical Literature]

[Patent Literature]

[Patent Document 1] Japanese Patent Laid-Open Publication No. 2004-254595

[Patent Document 2] Japanese Patent Laid-Open Publication No. 2006-028069

[Patent Document 3] Japanese Patent Laid-Open Publication No. 2007-181789

[Patent Document 4] Japanese Patent Laid-Open Publication No. 2006-219455

[Patent Document 5] Japanese Laid-Open Patent Publication No. 2012-176904

[Non-patent literature]

[Non-Patent Document 1] "J. Oleo Sci." vol.58, No.3, p133-140 (2009)

[Non-Patent Document 2] "Biochim. Biophys. Acta" Vol. 1558, No. 2, p161-170 (2002)

[Non-Patent Document 3] "Appl. Environ. Microbiol." Vol.69, No.5, p2595-2602 (2003)

It is an important issue for mammals, especially for humans, to impart activation or anti-aging. The industry continues to discover a variety of animal-derived, plant-derived activating substances or anti-aging agents, but in fact has not yet achieved sufficient and stable industrial use, and is constantly exploring new activators or anti-aging agents. An object of the present invention is to provide an activator and an anti-aging agent which are excellent in activation and anti-aging effects on cells, have safety sufficient for long-term use, and provide a cosmetic or quasi-drug which uses these as active ingredients. , pharmaceuticals, food and beverage products.

As a result of intensive efforts to achieve the above object, the present inventors have found that the above problems can be solved by the means described below, and the present invention has been achieved. That is, the present invention includes the following constitution.

A cell activator characterized by comprising a cellobiose ester.

2. The cell activating agent according to 1, wherein the cell activation system activates the skin cells.

3. The cell activator according to 1, wherein the cell activation system will Fibroblasts, hairy cells or dermal papilla cells are activated.

4. The cell activating agent according to 1, wherein the cell activation system promotes collagen production.

5. The cell activator according to any one of the above 1 to 4, which contains a cellobiose ester having a structure represented by the following formula (I) or (II):

(In the formula (I), R ₁ represents hydrogen or a hydroxyl group, n1 represents a carbon number of 12 or 14; in addition, R ₂ , R ₃ and R ₄ of the sugar 1 in the formula (I) represent an ethyl hydrazide group or a hydroxyl group)

(In the formula (II), R ₁ and R ₂ each represent hydrogen or a hydroxyl group; further, n1 represents an alkylene group having 11 or 12 carbon atoms, and n2 represents an alkylene group having 2 or 4 carbon atoms).

An anti-aging agent comprising the cell activating agent according to any one of the above 1 to 5.

A skin protectant comprising the cell activating agent according to any one of the above 1 to 5.

A skin roughness improving agent comprising the cell activating agent according to any one of the above 1 to 5.

A skin texture adjusting agent comprising the cell activating agent according to any one of the above 1 to 5.

A skin wrinkle prevention/inhibitor comprising the cell activating agent according to any one of the above 1 to 5.

According to the present invention, it is possible to provide a cosmetic, a quasi-drug (a skin external preparation, a bathing agent, a hair restorer, etc.), a pharmaceutical, a food or drink, and these are cellobiose esters as an active ingredient, and a cell viability excellent in safety can be expected. And collagen production.

Figure 1 is a graph showing the cell activation of CL using normal human skin fibroblasts.

Fig. 2 is a graph showing the promotion of collagen production using CL of normal human skin fibroblasts.

[activator]

In the present specification, "activation" means an expression intended to maintain or enhance cell function or cell activity. The result can help maintain cell function or cell activity constant and inhibit cell aging. Therefore, "activator" is synonymous with "cell activator" and has usefulness as an "anti-aging agent".

For example, the so-called "activation, anti-aging" of skin cells refers to the reduction of the function of skin cells caused by the accumulation of changes in the basement membrane due to aging or photoaging, or metabolic abnormalities, thereby preventing and improving the skin. Wrinkles, sagging, hardening, etc., while maintaining a youthful and healthy skin condition with elasticity. In addition, for example, for fibroblasts, it is intended to prevent the proliferation of fibroblasts caused by external factors such as ultraviolet rays, severe air drying, excessive skin cleansing, stress, smoking, or aging, and to prevent or improve the elasticity of the skin or Decreased tightness, wrinkles or sagging of the skin, etc. Further, for example, in the case of dermal papilla cells or hairy cells, it is possible to maintain the hair cycle and suppress hair loss by suppressing a decrease in function of dermal papilla cells or hairy cells due to aging, stress, and hormonal imbalance.

[Promotes collagen production]

In the present specification, "promoting collagen production" is an effect of increasing the amount of collagen produced by cells. At the junction of the epidermis and the dermis, collagen IV and VII are involved in the support of epidermal cells, and then. Due to aging or ultraviolet rays, especially ultraviolet rays, the collagen IV and VII are reduced, and the epidermal and dermal support functions are reduced. The selective function is weakened and the harmful components are easily affected by the skin, so wrinkles are formed. In addition, the extracellular matrix that constitutes the dermis is composed of fibrils in the dermis. It is produced by a cell, and is composed of a fibrous protein such as collagen or elastin and a polysaccharide called an acidic mucopolysaccharide such as hyaluronic acid, which directly relates to skin elasticity, metabolism, and vitality. If the activity of the fibroblasts decreases, especially due to aging or photoaging, the collagen biosynthesis is lowered and the modified elastin is increased. Further, the matrix metalloproteinase MMP-1 which decomposes the type I collagen which is a main component of the dermis is increased, and the decomposition of the extracellular matrix component such as collagen is increased, and damage occurs in the dermis, and the junction of the epidermis and the dermis is destroyed. And accelerate the decomposition of the dermis. Finally, the skin loses its elasticity and hydration, vitality, and produces wrinkles, fine lines, and rough skin, leading to skin aging. Therefore, the anti-aging effect by the "promoting collagen production" effect can be expected.

[cellosaccharide]

The cellobiose ester is a kind of bioester-produced glycol ester type biosurfactant, and the sugar ester type biosurfactant has interfacial activity and emulsifying ability. The term "biosurfactant" is a generic term for substances produced by biologically active interfacial activity and emulsifying ability. It not only exhibits excellent interfacial activity or high biodegradability, but also has various physiological effects, and thus may exhibit Different behaviors and functions of synthetic surfactants.

There are various variations in the structure of cellobiose ester (hereinafter sometimes referred to as CL), and the difference in molecular structure is based on the difference in the production microorganism. In the formula (I), R ₁ represents hydrogen or a hydroxyl group, respectively, and n1 represents 12 to 14. Further, R ₂ , R ₃ and R ₄ of the sugar 1 in the formula (I) represent an ethyl hydrazine group or a hydroxyl group. These are obtained in the form of a mixture, but it is also possible to carry out a purification separation operation using a separate CL compound. Further, in order to improve the water solubility, an alkali treatment may be carried out to prepare a sodium salt. Further, in the formula (II), R ₁ and R ₂ each represent hydrogen or a hydroxyl group. N1 means 11~12, and n2 means 2~4. These are obtained in the form of a mixture, which can be subjected to a purification separation operation using a separate CL compound, or a mixture can be used as it is. Further, in order to improve the water solubility, an alkali treatment may be carried out to prepare a sodium salt.

[Method of making cellobiose ester]

The cellobiose ester is obtained by extracting and purifying a culture solution of a cellobiose ester-producing bacterium. Examples of the CL-producing bacteria include microorganisms belonging to the genus Cryptococcus (genus Cryptococcus) or Ustilago, and having the ability to produce cellobiose esters. The microorganism of the genus Cryptococcus mainly produces the cellobiose ester of the above formula (I), and the microorganism of the genus Ustilago mainly produces the cellobiose ester of the formula (II).

The method for producing the cellobiose ester is not particularly limited, and a fermentation method for producing a microorganism using a known biosurfactant can be arbitrarily selected. For example, culture production of cellobiose ester can be produced by culturing Cryptococcus humicola or Ustilago esculenta by a conventional method. In addition to the above, the cellobiose producing microorganism may be used, such as Ustilago maydis, Pseudozyma flocculosa, Pseudozyma graminicola, or the like. The biosurfactant production microorganism is not particularly limited and may be appropriately selected depending on the purpose.

The fermentation base for producing the cellobiose ester may be a medium having a general composition, and the medium of the general composition includes an N source such as yeast extract or peptone. C source such as glucose or fructose, inorganic nitrogen source such as sodium nitrate, inorganic salt such as dipotassium hydrogen phosphate or magnesium sulfate heptahydrate.

Fermentation conditions such as pH or temperature, culture time, and the like can be arbitrarily set, and the culture solution after fermentation can be directly used as the biosurfactant of the present invention. Further, the fermentation solution after fermentation may be optionally subjected to any operations such as filtration, centrifugation, extraction, purification, sterilization, etc., and the obtained extract may be diluted, concentrated, and dried.

The inorganic nitrogen source is not particularly limited and may be appropriately selected according to the purpose, and examples thereof include ammonium nitrate, urea, sodium nitrate, ammonium chloride, and ammonium sulfate.

The method for recovering and purifying the cellobiose ester is not particularly limited and may be appropriately selected according to the purpose. For example, it can be recovered by centrifuging the culture solution and recovering the oil component, followed by extraction and concentration with an organic solvent such as ethyl acetate.

As the extraction solvent, an organic solvent such as water, an alcohol, a ketone, diethyl ether, dioxane, acetonitrile, an ester, xylene, benzene or chloroform may be used singly or in combination of two or more kinds thereof. The solution is mixed, and each solvent extract may be used in combination.

The extraction method is not particularly limited, and it is usually in the range of the boiling point of the solvent at normal temperature to normal pressure, filtered after extraction, or adsorbed, decolored, and purified using an ion exchange resin to form a solution, paste, or coagulation. It can be gelatinized or powdered. In many cases, it can be directly used in this state, but if necessary, it is possible to further increase the refining treatment such as deodorization and decolorization without affecting the effectiveness thereof. For the purification treatment such as deodorization and decolorization, an activated carbon tube column or the like can be used, and the usual means generally applied can be arbitrarily selected according to the extracted substance. get on. Refining can be carried out using a ruthenium tube column as needed, whereby a higher purity cellobiose ester can be obtained.

The cellobiose ester obtained in the above manner can also be used as an activator as it is, but it is preferably used in cosmetics, quasi-drugs, pharmaceuticals, and foods. The blending concentration of the cellobiose ester is appropriately selected depending on the degree of absorption, the degree of action, the form of the product, the frequency of use, and the like, and is not particularly limited, but is usually 0.00001% by weight to 0.1% by weight, preferably 0.0005% by weight to 0.05% by weight. More preferably, it is 0.0001% by weight to 0.01% by weight.

The use form of the cellobiose ester formulated in the activator is arbitrary. For example, the cellobiose ester can be directly used as an extract extracted from a culture solution, or a purified high-purity product can be used. Since the cellobiose ester has high hydrophobicity, it is preferably used by being dissolved in a nonionic surfactant or a lower alcohol or a polyhydric alcohol. Alternatively, a sodium salt of a cellobiose ester which is easily dissolved in water can also be used.

[skin protectant]

The fibroblasts are activated by using a cellobiose ester which is an active ingredient of the activator of the present invention. By normalizing the epidermal cells by activation of fibroblasts, the skin's barrier function and metabolic turnover are restored or restored, which can prevent or improve external factors such as ultraviolet rays, severe air drying, excessive skin cleansing, stress, and smoking. The effect of preventing or improving the skin elasticity or firmness caused by aging, and protecting the skin from wrinkles or slack.

[Skin Roughness Improver]

Fiber by using an active ingredient as an activator of the present invention A disaccharide ester that promotes activation of fibroblasts. With the activation of fibroblasts, the production of hyaluronic acid and the like as a natural moisturizing factor can be promoted, and the skin which is rough or dry can be improved.

More preferably, the activator of the present invention is prepared by disposing a cellobiose ester as an active ingredient in a cosmetic, a quasi-drug, a pharmaceutical, or a food.

[Skin Texture Conditioner]

The term "skin texture" refers to irregularities formed by depressions (skins) and ridges (skins) located on the surface of the skin. In addition, the term "skin texture" refers to a state in which the width of the skin groove is narrow and the skin ridge is flat and tidy, which is determined by the width and interval of the skin groove. These are composed of the thickness and hardness of the epidermis, the elasticity of the intercellular substance, and the like. When the constituent elements become uneven with aging or photoaging, the regularity of the texture is impaired. Collagen is involved in the elasticity of the intercellular substance. By using the cellobiose ester of the present invention as an active ingredient for promoting collagen production, the collagen production ability is activated, and the elasticity of the intercellular substance is improved, and the texture is adjusted to a normal state. By averaging the texture, it is possible to obtain an effect of improving the light scattering effect, disappearing the faint skin condition, and brightening the skin tone.

[Skin wrinkle prevention / inhibitor]

The term "skin wrinkle prevention/inhibition" refers to an effect of preventing wrinkles from forming with aging or photoaging. By using the cellobiose ester of the present invention, the collagen production ability of the cells is promoted, thereby preventing and suppressing wrinkles. Wrinkles are caused by the following causes: damage to dermal cells with exposure to ultraviolet light; or elastin due to aging The structure of the dermis, which is composed of collagen, is destroyed and leads to a decrease in the production of elastin or collagen.

[Use form of the present invention]

When it is carried out in the form of a cosmetic, a quasi-drug, or a pharmaceutical, it is preferably an external preparation. However, since the bio-surfactant can also be taken orally, it is not limited to the external preparation, and can also be used as an internal preparation or a food or drink.

The dosage form is not limited, and can be prepared into various dosage forms such as ampoules, capsules, powders, granules, pills, troches, solid agents, liquids, gels, bubbles, emulsions, creams, ointments, flakes, mousses, bathing agents, and the like.

Examples of the cosmetic, the quasi-drug, and the pharmaceutical include a base cosmetic such as an oral/exogenous pharmaceutical preparation, a lotion, an emulsion, a cream, an ointment, a lotion, an oil, and a dressing; a facial cleanser or a skin. Hair lotion, shampoo, moisturizing lotion, hair lotion, hair cream, hair oil, fog, hair styling agent, perm agent, hair growth agent, hair dye, hair growth / hair care agent, etc.; foundation, white powder , powder, lipstick, blush, eye shadow, eyeliner, mascara, eyebrow pencil, false eyelashes and other make-up cosmetics; nail polish and other cosmetic materials; perfume, bath, toothpaste, breath freshener / mouthwash , anti-odor, toilet, sanitary napkins, wipes, etc.

Examples of foods and drinks include beverages such as refreshing drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks; various types of healthy nutritional supplement foods, health functional foods, lozenges, capsules, beverages, and buccal preparations. .

The activator of the present invention is suitably applied to humans, but can be applied to animals other than humans in the case where various effects can be expected.

In addition to the cellobiose ester as an active ingredient, the activator of the present invention may be used in the form of cosmetics, quasi drugs, pharmaceuticals, foods and drinks, or may be used in combination as long as the effects of the present invention are not impaired. additive.

For example, oils and fats, waxes, mineral oils, fatty acids, alcohols, esters, metal soaps, colloidal and water-soluble polymer compounds, surfactants, vitamins, amino acids, whitening agents, Humectant, hair restorer, extract or extract of animal or plant or natural medicine, microbial culture metabolite, α-hydroxy acid, inorganic pigment, ultraviolet absorber, astringent, antioxidant, anti-inflammatory agent, sterilizing/disinfectant , hair agent, perfume, pigment/colorant, sweetener, nutrient enhancer, hormone, metal ion blocker, pH adjuster, chelating agent, antiseptic/mold inhibitor, cooling agent, stabilizer, emulsifier , animal/plant protein and its decomposition products, animal and plant polysaccharides and their decomposition products, animal and plant glycoproteins and their decomposition products, blood flow promoters, anti-inflammatory agents / anti-allergic agents, cell activators, keratolytic agents, wounds Therapeutic agent, foaming agent, thickening agent, oral agent, deodorizing/deodorizing agent, bittering agent, seasoning, enzyme, and the like.

Example

The invention is specifically illustrated by the following examples, but the invention is not limited to the examples.

[Example 1: Production of cellobiose ester (CL) using Cryptococcus humicola]

The colony of the cryptococci NBRC 10251 was inoculated into a seed medium (50 mL/500 mL long neck flask) to inoculate 1 Loop, and the inoculum culture was carried out. Incubate overnight at 27 °C. The obtained culture solution was used as an inoculum. The medium composition was as follows: 10 g/L glucose, 3 g/L sodium nitrate, 0.5 g/L magnesium sulfate heptahydrate, 0.03 g/L sodium dihydrogen phosphate, and 0.1 g/L yeast extract. In the culture system, 60 mL of the above-mentioned inoculum was inoculated into a production medium (6 L (10 L-jar)), and cultured at 27 ° C, 530 rpm (stirring rotation), and 3 L/min (air) using a 10 L bottle. The production medium was composed as follows: 10 g/L glucose, 3 g/L sodium nitrate, 0.5 g/L magnesium sulfate heptahydrate, 0.03 g/L sodium dihydrogen phosphate, and 0.1 g/L yeast extract. The culture solution cultured on the 6th day was centrifuged (7500 rpm, 20 min), and the cells were separated from the supernatant. After adding an equal amount of ethyl acetate:acetone = 4:1 mixture with the cells and stirring, the CL was extracted. The precipitate was separated into a supernatant and the supernatant, and the supernatant was dried by an evaporator to obtain a powder. In order to further remove the lipid impurities from the obtained powder, hexane (300 g) was added and stirred, and then the glass filter was filtered and evaporated to remove hexane. Thereby a mixture of CL is obtained.

[Example 2: Production of cellobiose ester (CL) using Ustilago esculenta]

The colonies of S. cerevisiae NBRC 9887 were inoculated with 1 loop in a seed medium (50 mL/500 mL long neck flask), and inoculum culture was carried out. Incubate overnight at 27 °C. The obtained culture solution was used as an inoculum. The medium composition was as follows: 50 g/L glucose, 3 g/L sodium nitrate, 0.5 g/L magnesium sulfate heptahydrate, 0.025 g/L potassium dihydrogen phosphate, 0.5 g/L dipotassium hydrogen phosphate, and 8 g/L yeast extract. In the culture system, 100 mL of the above-mentioned inoculum was inoculated into a production medium of 6 L (10 L bottle), and cultured at 27 ° C, 530 rpm (stirring rotation), and 3 L/min (air) using a 10 L bottle. Production medium The composition was as follows: 50 g/L glucose, 3 g/L sodium nitrate, 0.5 g/L magnesium sulfate heptahydrate, 0.025 g/L potassium dihydrogen phosphate, 0.5 g/L dipotassium hydrogen phosphate, and 8 g/L yeast extract. The culture solution cultured on the 6th day was centrifuged (7500 rpm, 20 min), and the cells were separated from the supernatant. After adding an equal amount of ethyl acetate:acetone=4:1 mixture with the cells and stirring well, the CL was extracted. The mixture was separated into a supernatant and a supernatant, and the supernatant was dried by an evaporator to obtain a powder. In order to further remove the lipid impurities from the obtained powder, hexane (300 g) was added and stirred, and then the glass filter was filtered and evaporated to remove hexane. Thereby a mixture of CL is obtained.

[Example 3: Sodium salting of cellobiose ester]

To the CL 4g obtained in Example 1 or 2, 20 mL of distilled water was added and stirred. To the solution, 1N NaOH was added dropwise to adjust the pH to 7-8. To pulverize it, the water was removed by an evaporator, and ethanol was further added and dried.

[Example 4: Structural analysis of cellobiose ester]

The CL obtained by Cryptococcus humicola and Ustilago esculenta was identified by 1H-NMR measurement (proton type nuclear magnetic resonance spectroscopy) at a resonance frequency of 500 MHz. For the measurement apparatus, the NMR apparatus AVANCE 500 manufactured by BRUKER Co., Ltd. was used, and the solvent was deuterated chloroform (CDCl3) and deuterated methanol (CD ₃ OD). As a result, it was found that all the powders obtained from the strains were CL. Formula (III) and Formula (IV) are as follows. Formula (III) represents the main structure of CL derived from Cryptococcus humicola, and formula (IV) shows the main structure of CL obtained from Ustilago esculenta. Regarding the respective representative structures, the peak of chloroform in the measurement using deuterated chloroform was set to 7.24 ppm, and the chemical shift attributable to the peak of the methyl proton of methanol in the measurement using deuterated methanol was 3.30 ppm. Shown in Table 1 and Table 2. Further, regarding Table 1, n1 = 14 and R ₁ = OH in the formula (III); with respect to Table 2, n1 = 12, R ₁ = OH, and R ₂ = OH in the formula (IV).

(In the formula (III), R ₁ represents hydrogen or a hydroxyl group, and n1 represents a carbon number of 12 or 14. Further, R ₂ , R ₃ and R ₄ of the sugar 1 in the formula (I) represent an ethylidene group or a hydroxyl group.

Formula (IV)

(In the formula (IV), R ₁ and R ₂ each represent hydrogen or a hydroxyl group. Further, n1 represents an alkylene group having 11 or 12 carbon atoms, and n2 represents an alkylene group having 2 or 4 carbon atoms)

[Table 1]

[Example 5: Cell activation using cellobiose ester of human normal skin fibroblast]

Normal human skin fibroblasts per hole of 2.0 × 10 ⁴ th embodiment seeded in 48-well microtiter plate. The seeding medium was used in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum. After incubating for 24 hours at 37 ° C and a carbon dioxide concentration of 5%, CL-Na salt was added to the test medium at a final concentration of 100 μg/mL to 1 μg/mL, and further cultured for 72 hours.

Distilled water was set as a solvent control. Then, 25 μL of the viable cell number measuring reagent SF was added and cultured for 3 hours, and the absorbance at 450 nm was measured using a microplate analyzer.

At the same time, the absorbance at 650 nm was measured as the turbidity, and the cell activation was evaluated based on the difference between the two measured values.

The evaluation results are shown in Fig. 1 in terms of the relative value when the cell activation activity of distilled water was set to 100.

According to Fig. 1, when CL-Na salt was added at a final concentration of 100 μg/mL to 10 μg/mL, cell normalization of human normal skin fibroblasts was shown to be higher than that of distilled water. In particular, when CL was added at a final concentration of 25 μg/mL, significant cell activation of 30% or more higher than distilled water was confirmed. According to the results, CL has an excellent cell-activating effect, suggesting that the application of CL to the skin improves the metabolism and regenerative ability of skin cells, and at the same time, it can effectively improve skin wrinkles, sagging, etc. due to aging or exposure to ultraviolet rays. .

[Example 6 Evaluation of collagen production ability by CL-Na salt]

The powder of the CL-Na salt described in Example 3 was dissolved in distilled water to prepare a final concentration of CL-Na salt of 100 μg/mL, 50 μg/mL, 25 μg/mL, 10 μg/mL, 5 μg/mL, and 1 μg, respectively. A /mL cellobiose solution was added to evaluate changes in the amount of collagen produced in normal human skin fibroblasts. Only distilled water was used as a control and evaluation was performed in the same manner. The evaluation was carried out in the following order. Normal human skin fibroblasts were seeded in a 48-well microplate at 1.0 × 10 ⁴ per well. The seeding medium was used in the addition of 1% fetal calf serum to Dulbecco's modified Igor medium (DMEM). After culturing for 24 hours at 37 ° C and a carbon dioxide concentration of 5 vol%, the cells were washed twice with PBS (-) (phosphate buffered solution), and then replaced with serum-free medium supplemented with any concentration of the sample, and further The culture was carried out for 3 days and 6 days under the same conditions. The type I procollagen C-terminal peptide produced by human fibroblasts was determined from the culture supernatant using a Procollagen type I C-peptide (PIP) EIA kit (TaKaRa). Procollagen type I carboxyterminal propeptide: PIP). Regarding the collagen production promotion rate, the standard was measured by the above ELISA kit, and a calibration curve was prepared based on the result, and the amount of collagen produced when the sample was added and the collagen when the sample was not added were determined based on the calibration curve. The amount of protein produced is used to determine the rate of promotion of collagen production.

According to Fig. 2, when CL-Na salt was added at a final concentration of 100 μg/mL to 10 μg/mL, collagen production of human normal skin fibroblasts was promoted to be higher than that of distilled water. In particular, when CL was added at a final concentration of 50 μg/mL, it was confirmed that a remarkable collagen production-promoting effect was 260% or more higher than that of distilled water. From this result, it is understood that CL has an excellent collagen production promoting action.

Collagen is produced by fibroblasts. However, the activation activity of the fibroblasts brought about by CL in Fig. 1 is 100 to 130%, and the collagen production rate by CL in Fig. 2 shows an extremely high value of 160% to 260%. Therefore, this result suggests that not only the ability to activate fibroblasts brought about by CL causes an increase in collagen production ability, but also promotes collagen production by acting on a certain channel of the metabolic system of collagen by CL.

[Example 7: Production Example of Beauty Liquid]

A cosmetic liquid of the composition shown below was produced by a conventional method.

[Example 8: Production Example of Emulsion]

An emulsion of the composition shown below was produced by a usual method.

[Example 9: Manufacturing Example of Cream]

A cream of the composition shown below is produced by a conventional method.

Purified water makes the whole amount 100

[Example 10: Production Example of Facial Cleanser]

A facial cleanser of the composition shown below was produced by a conventional method.

[Industrial availability]

According to the present invention, a cell viability, a collagen production-promoting action, and an anti-aging action which are excellent in safety from a biosurfactant can be expected, and a cosmetic or quasi-drug (such as an external preparation for skin) containing such an active ingredient can be provided. Bathing agents, hair restorers, etc.), food and beverages, and pharmaceuticals are also expected to be of great help to the industry.

Claims (10)

A cell activator characterized by comprising a cellobiose ester. The cell activating agent according to claim 1, wherein the cell activation system activates the skin cells. The cell activating agent according to claim 1, wherein the cell activation system activates the fibroblast, the hairy cell or the dermal papilla cell. The cell activating agent according to claim 1, wherein the cell activation system promotes collagen production. The cell activator according to any one of claims 1 to 4, which contains a cellobiose ester having a structure represented by the following formula (I) or (II): In the formula (I), R ₁ represents hydrogen or a hydroxyl group, and n1 represents a carbon number of 12 or 14; in addition, R ₂ , R ₃ and R ₄ of the sugar 1 in the formula (I) represent an ethyl hydrazide group or a hydroxyl group; II) In the formula (II), R ₁ and R ₂ each represent hydrogen or a hydroxyl group; further, n1 represents an alkylene group having a carbon number of 11 or 12, and n2 represents an alkylene group having a carbon number of 2 or 4. An anti-aging agent comprising the cell activating agent according to any one of claims 1 to 5. A skin protectant comprising the cell activating agent according to any one of claims 1 to 5. A skin roughness improving agent comprising the cell activating agent according to any one of claims 1 to 5. A skin texture regulating agent comprising the cell activating agent according to any one of claims 1 to 5. A skin wrinkle prevention/inhibitor comprising the cell activating agent according to any one of claims 1 to 5.