WO2004061115A1

WO2004061115A1 - PROCESS FOR PRODUCING SOLUBLE β-GLUCAN HAVING IMMUNOLOGICAL FUNCTION AND USE THEREOF

Info

Publication number: WO2004061115A1
Application number: PCT/JP2003/016979
Authority: WO
Inventors: Shu Uemura; Teruhisa Kaneko; Chihiro Kaise; Robert Nakamura; John Vercellotti; Sharon Vercellotti
Original assignee: Immudyne Japan Inc.
Priority date: 2002-12-27
Filing date: 2003-12-26
Publication date: 2004-07-22
Also published as: JP2004210895A

Abstract

It is intended to provide a process for effectively producing a soluble β-glucan having an immunological function and a skin cosmetic showing a remarkably excellent effect of preventing skin aging. A soluble β-glucan having an immunological function (soluble yeast beta 1,3/1,6-D-glucan, yeast and/or yeast extract) is produced with the use of β-glucanase and then employed in skin cosmetics.

Description

Description Soluble with immune function) 3-Klecan method and application

Shelf field

The present invention relates to a method of soluble-dulcan having a boat function and its use, and more particularly to a skin cosmetic having an excellent anti-aging function for skin and an S method of soluble / 3-glucan having an immune function suitable for use in the skin cosmetic.

Background

Glucan is a polysaccharide having glucose as a clean position. β— (1, 6) Branched) 3— (1, 3) Dalkan. Fermentation § ^ Other yeast species, mushrooms, plants and various bacteria; algae and algae (Chemi stry and Biol ogy of (1 → 3) — β-G 1 uc an s, BA St one and AE Clarke, 1992, reviewed by La Trobese University Press, Australia). β- (1, 6) branched) 3- (1, 3) dulcan has been shown to enhance immune function and cholesterol-lowering ability. Yeast combines at least three kinds of; 3-Dalcan; refractory 3-1, 3-D-Dalcan, rod-shaped / 3- 1, 6- D-Dalcan and β- (1, 6) branched ] 3— (1, 3) Glucan. However, linear—1, 3-D-glucan, linear; 3--1, 6-D-dalcan does not activate macrophages, NK cells or neutrophils, or very little. is there.

The iS— (1, 6) branched / 3_ (1, 3) glucan as a polysaccharide species is a fraction that binds to ¾1 by linking ill with glucose units and (1 → 6) 3-glycosylated linkage. Consists of buying. Yeast — (1, 6) branched / 3— (1, 3) dulcan is composed of several relatively short side chains (10% or 10%) fed by β- (1 → 6) glycosylated linkages. (1 → 3) / 3 glycosylated linkages are the members (90% of which are above) and are mostly composed of members. The chemical name of this dulcan is poly (1,3) —3—D-darcopyranosirile (1,6) -one-D-crecopyranose. There are a variety of different types of beta-glucans. Those members' thread! ^, Change with branch type, monomer type or substituent • It produces polysaccharides that exhibit very different physical and biological properties.

β-1,3 / 1,6—D-Dalcan is known to activate the invertebrate immune system as well as vertebrates. 6 _D—Dalcan is most effective in activating macrophages, NK cells and neutrophils. Yeast-derived; 3 glucan activates the immune system by binding to specific receptors on the cell membrane surface of macrophages. Activated macrophages increase the production of various cytokines as well as their phagocytic and bactericidal capabilities. Once the functions of macrophages and sputum cells are reduced, they can become responsible for the many beneficial effects of yeast iS glucan. For example, it is possible to improve the resistance to infections caused by infestation of bacteria, viruses, and protozoan parasites. In addition, macrophages are also considered to be bad if NK cells function. In contrast, the ability of the host can be improved.

Baker's yeast or brewer's yeast (brewer's yeast) as well as 3-1, 3/1, 6-D-glucan isolated from other yeasts are insoluble. In addition, / 3— 1, 3/1, 6-D-Dalcan is a side chain iS_ 1, 3/1, 6- D-glucan or i3—1, 3—D_Dalcan and i3—1, The decision to determine whether it is a mixture of 6_D—Krecans or all of these 3—Klecans is (at least 3 of these 3/3 as yeast disliked. —Creates all glucan) ^ Not J ability, but the solubility is getting worse due to the variability of the number of / 3-1, 6 side chains. Branched] 3-1, 3/1, 6- D-glucan only activates macrophages, so it is desirable to use a high yarn stringer / 3-1, 1, 3/1, 6- D-Dalcan . On the other hand, insoluble i3-1-1, 3/1, 6- D-glucan is difficult to formulate parenterally or topically administered crane IJ, and it is also difficult to absorb percutaneously. There are prescription restrictions. S¾i, a soluble glucan, has also been studied (Japanese National Publication No. 5-503952 and Japanese National Publication No. Hei 107-107702), but it is sufficient to have a soluble release or no immune function. In addition to these disadvantages, their use is limited to the medical use of wound healing. On the other hand, skin cosmetics containing soluble 3-glucan are known and the present applicant has already hesitated (Japanese Patent Application No. 2000-390953). However, the well-known cosmetic tolerability i3—Dalkan is not only difficult to formulate, but also prevents skin aging lh¾¾ (rough skin modification, keratin modification effect, There is a strong demand for the development of skin cosmetics that are easy to handle and have an excellent anti-aging effect on skin. Object of the invention

An object of the present invention is to solve the above-mentioned problems of the technique.

The first object of the present invention is to provide an effective method for producing soluble i3-dalton having immune function.

A second object of the present invention is to provide a dermatological agent that is easy to handle and has an excellent anti-aging effect on skin.

Disclosure of the invention

In the present invention, firstly, soluble iS-glucan having an immune function based on the use of 3-dalcanase in the process of elaborating 3-dalcan derived from yeast (soluble yeast bases 1, 3Z1, 6- D—Dalcan. Yeast and Z or yeast extract).

Secondly, the present invention includes a soluble / 3-glucan (soluble yeast, 1, 3/1, 6-D-Dal force, yeast and Z or yeast extract) that functions. It is in skin cosmetics that make it a tree.

DETAILED DESCRIPTION OF THE INVENTION

In the present invention, it is synonymous with “soluble ttJ” and “solubility” and means that it dissolves in water at room temperature (including colloidal form) on the shelf. The difficulty of shelf may be 1% by weight of the cosmetic. For example, the obtained β-glucan-containing substance is dissolved (dispersed) completely in water or colloidally at a concentration of 1% by weight at room temperature. [Typically obtained] Soluble yeast that enhances immune function in 3-glucan-containing products / 3-1, 3/1, 6-D-Dalton content is 95-; L 0 0% by weight This soluble yeast i3_l, 3/1, 6-D-Glylecan preferably has a content of 2500-600.

The koji method of the present invention is sensitive to the use of | 3-Dalkanase as a hydrolysin in the process of purifying yeast-derived koji (insoluble '14) 3-glucan.

The yeast (including yeast extract) that produces purified i3-Dalcan can be used. Specifically, live yeast cells themselves, dried yeast powders and Examples include soluble yeast extracts obtained by solubilizing, extracting and purifying yeast cells under appropriate conditions such as autolysis, hot water extraction, mi iY, and enzyme.

Yeasts include yeast jS-1,3 glucan, yeast 0-1,3 / 1,6-D-glucan, and genus Saccharomyces such as brewer's yeast, baker's yeast, and obtain yeast extract. Physics, 匕 ^ 0 勺, and biochemical methods can be used to break down the crumbling fungus. In the present invention, it is possible to select and shelf regardless of the fungus used as a raw material, the origin, the extraction method, the koji, or the S method of treated wisteria. Among them, commercially available yeast dry powder, yeast extract is easy to handle and readily available 蟒 (J. Above fermentation yeast extract usually contains protein, lipid, m organic acid, vitamin, amino acid, Nucleic acid-related quality is included.

Yeasts suitable for use in the present invention are i3-1-3 glucan, β-1, 3/1, 6-D-glucan having a molecular weight of 2500 to 400,000 or less, preferably 2500 to 60,000 koji, and beer fermentation. Examples include mothers, baker's yeast, and other genus Saccharomyce s, and J is 95% or more.

A typical example of the i method of the present invention will be described below.

After the fermented starch is treated under basic conditions, the insoluble '1' powder is washed with water and then treated with acid. In addition, the powder is treated with 3-Dalcanase. Alternatively, the fermented rice cake moon cake is treated with acid, washed at 7K, and then treated with sodium sootate. Wash the powder and treat with / 3-Dalcanase. Centrifuge the enzyme-treated mixture and remove the enzyme-treated powder. The supernatant is filtered through a membrane with an apparent cut capacity of 100,000 daltons and separated into a high "volume and low ^ *. After this, the substance is filtered through a membrane with 1000 daltons ^? 4, • Precipitate β-1,3 / 1,6-D-glucan and wash with absolute alcohol before drying in a vacuum oven.

Soluble j8—1, 3/1, 6-D-Dalcan that immunizes immune functions is S¾t from yeast or other microorganisms containing / 3—1, 3/1, 6-D-glucan as follows be able to.

The t process described here relates to the production of ίδ ^ ΐ * soluble that retains macrophage activation ability (immune function)) 6-1, 1, 3/1, 6- D-crecan. ： ^ Solubility to machine / 3— 1, 3/1, 6— D-Dalkan cocoons are the first component of / 3-Dalcan Threads containing alkali and acid soluble / 3-1, 3/1, 6-D-glucan (composed of fermented Tsuruta cysts yeast wall)! ! Process Xiemoto. This allows for the action of the enzyme; 3-Dalkane is obtained. After this treatment, the dissolved; 8-1,3X1,6-D-Dalcan is purified by enzymatic hydrolysis with; 3-Dalcanase. Here, the impurity β-glucan is removed.

The above manufacturing process is summarized as follows.

a. The fermented tubule or fermented rice bud wall does not solubilize / 3—1, 3,1,6-D—glucan; To do.

b. After washing 7, acid-treat the soot containing β-1 ′ 3/1, 6-D-Dalcan under heating. Alternatively,

a-1.

b-1. After washing with water, treat the powder containing β-1, 3/1, 6-D-grecan with sodium acid.

c After washing with τ, add the powder in b or b-l with dulkanase from yeast.

d. Centrifuge the mixture and strain the powder.

e. Filter the supernatant containing the enzyme / 3—glucan through a membrane with an apparent ¾¾t capacity, eg, 100,000 daltons. Next, treat the 3-dalcan (under control) as in g below to obtain colloidal 1, 1, 3/1, 6-D-dalcan.

f · Pass the eluate through a membrane with an apparent cutting ability, eg 1000 Daltons. And ii; ^ Sorry is treated as follows to obtain soluble) 3-Dalcan.

g. Add to e from ί and suspend in 100% ethanol.

h. Wash items with ethanol and comfort. If necessary, it can be further purified and fractionated by filtration through a membrane of H with a different cut capacity or by chromatography.

Disperse the whole fermented rice cake or fermented fan rice cell wall in the alkaline night and heat it. As the alkali τΚ solution at that time, an alkali metal or alkaline earth metal hydroxide, for example, sodium hydroxide or potassium hydroxide of 0.05 to 1 ON is preferably used. This step is performed at 4 ° C to 150 ° C, but preferably at about 80 ° C. Also, the process is always ij, or under pressure, with about 15 Pascal Z1 2 rc being particularly preferred. The processing time is usually 10 minutes to 48 hours, but it depends on the type of alkali ^^ and ffi iK. Once the alkali treatment is performed, the powder is separated from the aqueous solution by an appropriate means such as centrifugation, etc. 1 ". This alkali treatment may be repeated once or several times. Wash the mulberry cell wall once or several times, then treat with acid such as «, formic acid, acetic acid, etc. ρ Η = Acidic acid of 1 ~ 5, usually 4. (： ~ 1 5 0 ° Treat with C for 15 minutes to 48 hours, especially 3% vinegar 酉 τΚsolute ^ 85 ° C, preferably for 45 minutes and extract.Insoluble raw material (glucan) is a suitable process Remove TT from the solution by washing with water once or several times and then re-it ^ using appropriate means.

The residue is then washed with 15-75% sodium hypochlorite at 4 ° (from 15 ° C to 15 ° C for 15 minutes).

4 Remove for 8 minutes. Wash the remainder with water once or several times.

Treat the powder with β-Dalkanase, usually from 10 ° C to 80 ° C for 15 minutes to 48 hours. This condition depends on ^^, extreme night advantage, and enzyme ¾. The soluble enzyme digestion is separated from the undigested material by an appropriate method such as centrifugation or ¾¾1. Soluble extinguishment [^ Fee! ^ Conclusions, fractionation by binding method.

Thus, a soluble, ίδ ^ ΐΜ (2500 to 60,000)] 3-1, 3/1, 6_D-glucan having a function of ^ ¾ can be obtained. The conventional S8t ^ method has the problem that about 16% mannose and the like and impurities (cannot show transport function)) contain 3-Dalcan. Soluble yeast) 3-1, 3/1, 6-D-glucan is 9

It became a powerful ability to obtain soluble; 3-glucan containing 5% or more of high thread cake.

Next, the skin cosmetic of the present invention will be described.

The skin cosmetic of the present invention contains soluble | 3-Dalcan (yeast and Z or yeast extract) as an essential component, preferably as a lipozo »and Z or sea? It is prepared by blending ingredients commonly used in soaking water and skin cosmetics.

The form of the skin cosmetic of the present invention is not particularly P-armed. For example, skin care products such as emulsion, cream, chemical ffi7_K, packs, facial cleansers, massages, body cosmetics and emulsifying fans, etc. Products and quasi drugs. Both are water-based That's right. The dosage form of cosmetics is also arbitrary. For example, it is possible to take a solubilizing emulsifier form or a β form.

In the skin cosmetic of the present invention, the soluble / 3-glucan may be added in an amount of 0.001 to 1%. Soluble / 3—glucan distribution * is less than 0.0 0 1%, the immune activity improvement effect unique to soluble i3_Dalcan is not exerted in ¾1, and when it exceeds 1%, discoloration, May affect the stability of the system, which is not desirable.

Examples of ingredients to be formulated as (optional) ^ in the skin cosmetics of the present invention include oils, powders, surface active substances, low alcohol, high liver compounds, gelling agents, UV absorbing UV scattering agents, antioxidants There are components commonly used in ordinary cosmetics such as lh dyes, preservatives, fragrances, and cosmetic ingredients, and these can be selected and used as long as the effects of the present invention are not impaired.

Liposome membrane shape / ¾ base material is natural or synthetic such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, lysophosphatidylcholine, sphingomyelin, egg yolk lecithin, ^ a lecithin Phospholipids or silicon-added phospholipids, cholesterols such as cholesterol, cholesterol alkyl esters, phytosterols, phytosterol alkyl esters, glyceglycoglycolipids, asildarcosides such as cetylgalactoside, dialkyl-type synthetic surfaces Active I4, N-acylsphingosine or its sulfate ester, N-acylsphingoglycolipid, N-higher acyl dirtathion or a mixture of tt ^ S or more It is below. In addition, decorative alcohols, higher alcohols, high and full fatty acids, and the like can be added as necessary to stabilize ribosomes or improve phase transition.

Sea extinguisher water is a natural water rich in nutrients that is low in temperature, clean, and rich in nutrients. The water intake depth is 2500-0500m, preferably 3200-0400m. Since deep ocean water contains numerous minerals, it has excellent moisture retention. ¾Sf is excellent in raw life, which is thought to promote penetration of ingredients contained in cosmetics into the skin. The distribution of the seabed water is usually 40 to 99%, preferably 50 to 60%.

The preferred optional self-synthesized component will be described more specifically. Examples of the fats and oils include jojoba, oil, castor oil, olive oil, oil, coconut oil, palm oil, cacao oil, mink oil, turtle oil and the like.

Examples of hydrocarbons include I paraffin, petrolatum, microcrystalline wax, and squalene.

Examples of waxes include beeswax, lanolin, carnauba wax, and candelillaro. Examples of lunar fatty acids include myristic acid, palmitic acid, stearic acid, oleic acid, isostearic acid, lauric acid and the like.

Synthetic esters include isopropyl myristate, isopropyl palmitate, butyl oleate, myristyl myristate, octyldodecyl myristate, propylene glycol monostearate,? Examples include L myristyl L, isostearyl malate, glyceryl monostearate, and distearyl dimethyl ammonium chloride.

Oils, hydrocarbons, waxes, lunar fatty acids, synthetic esthetics are usually combined. It is blended at a ratio of ~ 5 Ow / w%.

Examples of alcohols include ethanol, 1,3-butylene glycol, propylene glycol, lauryl alcohol, cetanol, stearyl alcohol, and oleyl alcohol.

Alcohols are usually blended at a ratio of 0 to 1 OwZw%.

Surfactant 'I4 ^ | J includes glycerin fatty acid esters, sorbitan fatty acid esters, polyoxyethylene fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene alkyl ethers, coconut oil fatty acid monoethanolamide , Polyoxyethylene hydrogenated castor oil, sodium lauryl sulfate, polyoxyethylene darericyl pyrostearate, sodium alkylbenzene sulfonate, polyoxyethylene stearyl ether, dialkylsulfosuccinic acid, cetylpyridinium bromide, n-o chloride Decyltrimethylammonium, monoalkyl phosphate, N-acylsulfuric acid, N-acylglutamine, sucrose fatty acid ester, polyoxyethylene sorbitan monostearate And polyoxyethylene lauryl ether sodium sulfate, polyoxyethylene and lanolin. The surface activity '14 ^ 1 is usually added at a rate of 0 to 2 w / w%.

Examples of the thickener include strong lpoxyvinyl polymer, methylpolysiloxane, dextran, strong lpoxymethylcellulose, carrageenan, and hydroxypropylmethylcellulose.

The thickener is usually blended at a ratio of 0 to 1 wZw%.

Examples of the humectant include glycerin, propylene glycol, 1,3-butylene glycol, puffer glutamic acid, acetyl tiltamic acid, hyaluronic acid, procyanidine and the like. Moisturizer is usually. It is blended at a rate of ~ 5 w / w%.

Examples of preservatives include benzoic acid, salicylic acid, dehydroacetic acid or salts thereof, phenols of parabenzoic acid esters, and tricky sanhalocarban.

The preservative is usually blended at a ratio of 0 to 0 · 3wZw%.

Any fragrance may be used as long as it is generally used for cosmetics.

Examples of pigments include iron oxide, titanium dioxide, acid bffi-lead, kaolin, and talc. The prescription is usually blended at a rate of 0 to 1 wZw%.

觀 IJ includes: Wheat oil, Vitamin A, Vitamin B 2, Vitamin E, Ascorbile acid mono-2-magnesium or sodium phosphate, D-Pantole alcohol, Dipotassium glycyrrhizinate, Dartathione, UV absorber, Chelating agent, Examples include plant extracts and microbial metabolite Z extracts.

W, usually 0 to 3 w / w%.

As water for water, tap water, mineral water, brine, seawater, ultrapure water,

¾- ^ purified water and the like. Water is blended in an arbitrary ratio.

Yeast, yeast extract, especially yeast cell wall —Dalcan has a high texture of ^ mi kinetic I ', so various cells and activities related to immune enhancement such as T cells and complement system It has been reported that it has a slight biological activity, while it has a brilliant biological activity, but on the other hand, there is almost no horn | However, epidermal Langer-octance cells play a major role in skin immunity, and have the presentation function of transmitting information to sputum cells when there is an external source. Therefore, it is necessary to mix these cells with cosmetics. Unlike the prevention of aging through care centering on »», it shows an improvement in the immune function of the biological system itself. In addition, it is possible to improve the skin immunity function once by blending these with sea layer water that has a high degree of skin property, as well as ingredients that have anti-oxidant power and moisturizing and soft ingredients. ^ = Kiru.

«Skin aging prevention cosmetics act mainly on rough skin (due to seasonal changes) and skin trauma caused by temporary deterioration of skin function. Drowning due to systemic malfunction requires an improvement in the balance of the person's entire biological system. The skin cosmetics containing S-Dalcan (P-P) of the present invention acts on the wrinkle system and improves the balance of the biological system from the inside of the skin function. Of course, the effect on the human body is very sensible.

The skin cosmetics of the present invention have been used poorly solubilizing monoglucans that could not be formulated with water-soluble and transparent dosage forms of aqueous cosmetics (for example, chemicals can be formulated, and cosmetic cosmetic dosage forms The ij ability is used regardless of the choice, and is excellent in wrinkling on the skin, and exhibits excellent skin aging prevention effects on wisteria.

Furthermore, by using a low β-glucan, the rate of liposomes is dramatically improved, and it becomes possible for the first time to prepare a ribosome carrying more β-dulcan. As a result, the amount of iS-Dalcan on the skin? ! Improves MS MS and gains higher skin care effects (moisturizing, acupuncture, wrinkles, skin turnover, etc.).

Example

EXAMPLES Next, the present invention will be illustrated by examples, but the present invention is not limited to these examples.

Example 1:

Preparation of soluble iS-1, 3/1, 6- D-Grecan from the fermentation

To the fermenter, 10 times the amount of 1.5 N sodium hydroxide was added with stirring, and the mixture was then heated at 60 ° C. for 30 minutes. The heated mixture was autoclaved for 15 minutes at 15 psi and 12 C conditions. Thereafter, the heated mixture was cooled and centrifuged to separate the supernatant from the powder. The residue was washed with zK while stirring with 10 times the amount of water for 15 minutes. After separating the supernatant by centrifugation, add 10 times the amount of 3% vinegar ¾_Κ 謹 at 37 ° C, and add 4% at 85 ° C. Stir for 5 minutes. Thereafter, the mixture was separated, and the supernatant was separated from the powder by centrifugation. The / 3-1, 1, 3/1, 6-D-glucan powder from this concession was washed with water with stirring for 15 minutes. The fermented crane was treated with 10% 3% acetone in water at 37 ° C, koji was added, heated to 85 ° C, and stirred for 45 minutes. After washing the residue with water, it was treated with 15-75% of hypochlorous acid: * sodium acid. (Preferable conditions are a 20% concentration, a temperature of 4 to 150 ° C., more preferably 50 ° C. for 15 minutes to 48 hours.) The straw was washed several times.

Crude (insoluble / 3-1, 1, 3/1, 6- D-glucan was dispersed in 3 times the amount of water and sprinkled into 40 to 20 to 25 times the amount of water. From NovoNard i sk, Far nk 1 it on, NC 'Purchased V isco zyme L-β-Ι, 3/1, 6— D —Darkanaize (B i oc ata lys ts Ltd. Pon ty ri dd, 100 ml of TP654 / 2 P or other 3 / 3-dalcanase from UK) was added and stirred at 80. The suspension was then threaded with a filter to cut 100000 daltons Separated through a filter, repeated for 10 hours, always passing water at 80 ° C

(Adding) By keeping the inside of the reaction tube constant. The filtrate was collected over 10 hours. This was used to prepare colloidal 0-1, 1, 3/1, 6-D-glucan. The amount of sugar to be reduced was confirmed in the sample collected every liter. The filtrate was fractionated through a hollow fiber that cuts 1,500 ^ Τ * and dialyzed against salt, low-color pigment, and sugar. Dialysis was performed by pumping water through the outer hollow fiber at a water speed of 800 ml / m i η. And after 10 temples, 14.5 liters of food were collected. The hopping filtrate was concentrated with a reverse osmosis membrane and then filtered with a rotary evaporator. Β-1, 3/1, 6-D_Dalcan was obtained as a product from a food product by exempting the filter material with 10 times the amount of 95% ethanol for one day.

The material was collected, washed with 95% ethanol and dried. (Completely soluble jS with cocoon function — 1, 3/1, 6-D—glucan: ^? ¾3, cloth 4141 to 250208, average cocoon weight = 47 457 daltons, dispersion rate = 1.43. Of these, 75% It was ^ ¾ in the range of 37121 to 83902 Dali Reton) 'Using 3: 1, 3/1, 6-D-glucan 10 times the amount of 95% ethanol from the theory of membranes with cutting ability of ΛΙ00000 And stay at low temperature all night long It was allowed to settle from the outer filtrate. Collected porridge, washed with 95% ethanol, dried (colloidal β-glucan ;: ^ cloth 13148-473616 Taleton, average: 1474 20 Dalton, scatter rate = 1.90. Of which 75% is 91510- 256863 of Taleton range :).

The dried precipitate is further purified by chromatography (affinity, dialysis, chromatography, etc.) or by dialysis using a filter that can select various ¾.

Completely soluble / 3— 1, 3/1, 6— D—Shows the physical properties of Dalkan.

Soluble / 3-1-1, 3/1, 6-D-glucan properties with immune function

^ Μ: 12000—60,000 talileton

Soluble; 2.5 gm / ml

Underwater color; light beige

¾¾¾ at 280 nm; 2632

PH 7.0 of 1% aqueous solution

β-1,3 bond% 83

β-1, 6% bond 17

β-l, 4% None (not measurable)

Example 2:

The soluble β-Dal force that has a fine function prepared by the method of Male Example 1.

The result of making ¾τΚ containing 0.05% Taleton) in the morning is 8 weeks on the face of 100 women. The present test was hired specialists as the ^ ¹ is the result of Hosohito was monitored scratch mosquitoゝfelt how after shelf

(Formulation example. 1) Yeast extract (soluble—Dalcan;

Liver volume approx. 50,000 Tareton) 0. 05

Red wine yeast extract 0.1

Ridge water (reverse osmosis membrane) 50. 0 Lecithin 1.0

Cholesterol 0.5

Age Rain Acid 0.5

Glycerin 3.0

1, 3-butylene glycol 7.0

L—Arginine 0.25

Saturated fatty acid glyceryl 1.0

Glycerin fatty acid ester 1.5

Higher alcohol 0.5

雜 Alcohol fatty acid ester 0.5

Moisturizer 0.5

Preservative 0.5

Adjust to 100

® The method was prepared according to a conventional method and used as an emulsion.

The seawater layer water used here (reverse consideration) is the water that has been treated by retreating the ocean.

Achievement improvement rate (%)

28 days later (4 weeks after 56 days (8 weeks ^)

64% 80%

Skin tension 45% 68%

Skin wrinkles 22% 66%

As can be seen from the above results, more than half of the people after about 4 weeks of use have an effect of improving the moisturizing feeling of the skin. It was found that tampering effects were observed.

In addition, more than half of the skin was improved by turning over the skin for 8 weeks, and a considerable improvement was observed in the skin wrinkles.

Example 3: Soluble glucan prepared by a method according to the method of Example 1 from about 800 to 100

0% Dalton) was prepared, and it was used on women's faces in the morning and evening for 8 weeks. The skin wrinkled condition was visually observed and the degree of improvement was graphed. The results are shown in Figure 1. (In addition, is as shown in male example 2.)

As can be seen from the graph, it was found that the wrinkles in the skin were improved by the fineness of 8 weeks.

Example 4:

A milk containing 0.1% 5% soluble dull force prepared by the method according to the method of Example 1 was prepared. Fig. ² shows the results of examining the skin surface condition in a replica after 8 weeks of use and examining the improvement rate. (In addition, is as shown in Example 2.)

In any case, it exists diagonally confirmed before use f i n e w r

It was found that the line of 1 n k 1 e became almost inconspicuous after 8 weeks, and that the skin's protective effect was improved and that f i n w r i n k 1 e became inconspicuous.

Case 5:

An emulsion containing 0.05% of soluble 'I raw klecan ½H¾ approximately 5 0 0 0 0 evening lens) prepared by the method of Example 1 was prepared. Fig. 3 shows the results of a tape stripping test on the condition of the stratum corneum of the skin after using it for 8 weeks in the morning and evening, and examining the improvement rate. (The prescription is as shown in Example 2.) From the results of tape stripping, keratinocyte coverage was recognized before conversion in any of the cases, but 8 weeks after use. It turned out to be almost inconspicuous, and the stratum corneum was improved to a healthy state.

It should be noted that the more keratinocytes are present; i¾T indicates delamination of the stratum corneum, indicating that the skin condition is not healthy.

Example 6:

Soluble dulcan prepared by a method according to the method of Example 1 about 5 0 0 0 0 dart Was prepared on the upper arm of 10 females for 4 weeks, and the skin turnover state was tested by the tape stripping method. Figure 4 shows the results of the improvement rate. (The prescription is as shown in Example 2.) As shown in the result of microscopic examination of the stratum corneum collected by tape stripping, the stratum corneum was peeled off in the state where the skin was previously roughened (color). In the case where nothing is applied after that, the skin cell state further deteriorated, and even the cell nucleus was seen as black spots, but it was soluble) 3) Glucan included It was found that by applying the latex, flfT improved the skin condition of the stratum corneum and brought it closer to a healthy skin condition.

Brief Description of Drawings

Fig. 1 is a graph showing the wrinkle improvement results (visually) of Example 3.

Fig. 2 is a graph showing the wrinkle improvement results (crane by replica) of Example 4.

FIG. 3 is a graph showing the improvement results of the stratum corneum in Example 5.

FIG. 4 is a draaf showing the improvement result of Kakuda rice cyst of Example 6.

Claims

The scope of the claims

1. Soluble jS-glucan with an immune function based on the use of / 3-Dalcanase at the level of ß-Dal force of yeast-derived (soluble yeast, 1, 3Z 1, 6-D —Dalkan. Yeast and / or yeast extract) method.

2. A skin cosmetic composition that contains a soluble / 3-glucan (soluble yeast beta 1, 3/1, 6-D-glucan. Yeast and ζ or yeast extract) having an immune function.

3. The skin cosmetic according to claim 2, which is a water skin cosmetic.

4. The skin cosmetic according to claim 2 or 3, which is in the form of a liposome dispersion.

5. The skin cosmetic according to claim 2, further comprising deep ocean water.