JPH10245316A - Agent for preventing and ameliorating aging of skin - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject new cosmetic by compounding a phosphorylated polysaccharide having humectant action and collagen-producing performance as an active component. SOLUTION: This cosmetic contains a compound of the formula (Glc is glucose residue; Gal is galactose residue; Rha is rhamnose residue; (m) is 0-3; (n) is 1,000-5,000) as an active component. The component of the formula can be produced by hydrolyzing actinase E of defatted milk, treating the defatted milk with ultrafiltration membrane, sterilizing the filtrate to obtain a culture medium, aseptically pipetting the sterilized medium into a jar fermenter, inoculating a precultured liquid, culturing at 20 deg.C and pH5.5 using NH3 water as a neutralizing agent, subjecting the cultured product to centrifugal separation after culture to recover the supernatant, adding an equal amount of CH3 OH, recovering the produced precipitate, dissolving the precipitate in 0.2N saline water, repeating the precipitation operation with CH3 OH, subjecting the treated precipitate to electrophoresis, recovering the component impregnated into the gel and purifying the recovered fraction by ion-exchange chromatography to obtain purified polysaccharide. It is necessary to compound a cosmetic with >=0.001wt.% of the compound of the formula.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an agent for preventing and improving skin aging comprising a phosphorylated polysaccharide as an active ingredient. The agent for preventing and improving skin aging of the present invention is used as cosmetics.

[0002]

2. Description of the Related Art Normally, the skin has a water-absorbing ability and a water-retaining ability, so that its flexibility and elasticity are maintained. However, when these functions decrease due to aging, the water-soluble fraction of the skin is lost, It is believed that the skin surface dries, resulting in a so-called "skin roughness". The main components of skin are
Collagen, elastin and the like are known, and it is said that these components contribute to skin flexibility and elasticity. Also, these functions are reduced by aging,
It is thought to be caused by a decrease in collagen metabolism and a decrease in hyaluronic acid having a moisturizing ability existing between cells. Accordingly, cosmetics containing collagen, hyaluronic acid and the like for the purpose of improving these aging skins are commercially available.

[0003] On the other hand, it is known that various lactic acid bacteria produce polysaccharides. In addition, Streptococcus lactis or Lactococcus lactis
It has been reported that some strains of lactic acid bacteria such as lactis (Lactococcus lactis ), Streptococcus cremoris (Streptococcus cremoris) or Lactococcus cremoris (Lactococcus cremoris ) produce phosphorylated polysaccharides (Japanese Unexamined Patent Publication No. No. 229702; Nakajima et.
al., Carbohydr. Res., vol.224, pp.245-253, 1992).
Furthermore, Lactobacillus sake (Lactobacillus sake )
It has been reported that some strains of lactobacilli also produce phosphorylated polysaccharides (WO94 / 12656). In each of these phosphorylated polysaccharides, monosaccharides such as glucose, galactose and rhamnose repeat a certain sequence to form a sugar chain, and a phosphate group with a monosaccharide or a glycerol group as a side chain thereof is It differs from neutral polysaccharides in that it has a structure linked directly or via another monosaccharide. In addition, among the three oxyacids of the phosphate group, two are involved in an ester bond with a monosaccharide, but the remaining one is in a free state, and thus can participate in a reaction with another substance. It has the property of

[0004]

The present inventors have made various studies on the utility of phosphorylated polysaccharides, and found that the phosphorylated polysaccharide has a moisturizing action and collagen-producing ability. Was. Then, it has been found that by blending this phosphorylated polysaccharide as an active ingredient of a skin aging preventive and ameliorating agent, the effect of preventing and improving skin aging can be imparted to cosmetics and the like, and the present invention has been completed. Therefore, an object of the present invention is to provide an agent for preventing and improving skin aging containing a phosphorylated polysaccharide as an active ingredient.

[0005]

SUMMARY OF THE INVENTION The present invention is an agent for preventing and improving skin aging, comprising a phosphorylated polysaccharide as an active ingredient. The phosphorylated polysaccharide which is an active ingredient of the agent for preventing and improving skin aging according to the present invention is a known compound as described above (JP-A-3-229702; Nakajima et al., Carbohyd
r. Res., vol. 224, pp. 245-253, 1992; WO 94/12656), for example, a compound represented by the following structural formula.

[0006]

Embedded image (Where Glc represents a glucose residue, Gal represents a galactose residue, and Rha represents a rhamnose residue, respectively.
In addition, the numbers written in large letters in the formula represent each binding site, m is an integer of 0 to 3, n is a repeating unit,
Shows 1,000 to 5,000, respectively. )

[0007]

Embedded image (Where Glc represents a glucose residue, Gal represents a galactose residue, and Rha represents a rhamnose residue, respectively.
In the formulas, the numbers written in large letters indicate the respective binding sites, and n indicates the unit of repetition, and indicates 1,000 to 5,000, respectively. )

[0008] Streptococcus lactis (S
treptococcus lactis) or Lactococcus easy <br/> lactis (Lactococcus lactis), Streptococcus click <br/> Remorisu (Streptococcus cremoris) or Lactococcus cremoris (Lactoco ccus cremoris) lactococcal and Lactobacillus sake, such ( Lactobacillus such as Lactobacillus sake ) is cultured, and a phosphorylated polysaccharide can be obtained from the culture. The agent for preventing and improving skin aging of the present invention is used in the form of cosmetics such as lotions and creams.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, the above-mentioned phosphorylated polysaccharide is used as an active ingredient of an agent for preventing and improving skin aging. The agent for preventing and improving skin aging of the present invention is preferably used as an emulsifying system, a solubilizing system, a spray, an aerosol, etc. which are common forms for cosmetics. And the skin aging preventive and ameliorating agent of the present invention is directly applied to a part of the skin such as the face or hands that causes rough skin due to drying,
An appropriate amount may be applied at least once a day.

As the phosphorylated polysaccharide used as an active ingredient of the agent for preventing and improving skin aging of the present invention, not only a purified phosphorylated polysaccharide but also a culture of a lactic acid bacterium producing the phosphorylated polysaccharide is used as it is. Alternatively, a precipitate formed by adding a solvent such as ethanol to a culture of a lactic acid bacterium producing a phosphorylated polysaccharide may be recovered and used. These phosphorylated polysaccharides can be easily incorporated into cosmetics in liquid or powder form as skin aging preventive and improving agents.

In order to exhibit the effect of preventing and improving skin aging by incorporating the agent for preventing and improving skin aging of the present invention into cosmetics and the like, phosphorylation of at least 0.001% by weight or more, preferably 0.01 to 20% by weight. It is necessary to blend a polysaccharide into cosmetics and the like. In addition, the skin aging preventive and ameliorating agent containing the phosphorylated polysaccharide of the present invention as an active ingredient includes substances such as ascorbic acid and derivatives thereof which are conventionally known to have a collagen production promoting action and a melanin production inhibitory action. In combination with cosmetics and the like. It has been confirmed that the phosphorylated polysaccharide used as an active ingredient of the agent for preventing and improving skin aging of the present invention has no toxicity or irritation to the skin and no side effects.

Next, reference examples, examples and test examples will be described.
The present invention will be described in detail.

Reference Example 1 A phosphorylated polysaccharide was obtained according to the method described in JP-A-3-229702. That is, skim milk completely hydrolyzed with actinase E (manufactured by Kaken Pharmaceutical Co., Ltd.) was treated with an ultrafiltration membrane, and the retentate was used as a medium. The medium was sterilized, and 9.8 L of the medium was aseptically dispensed into a 10 L jar fermenter. And Streptococcus cremoris SBT-0495
After inoculating 5% of the precultured solution of (FERM P-10053), ammonia water was used as a neutralizing agent, and the pH was maintained at 5.5 to obtain 20%.
The culture was performed at 24 ° C. for 24 hours. After completion of the culture, the culture solution was centrifuged to collect the supernatant, and an equal amount of ethanol was added to form a precipitate. The precipitate was collected. Further, an operation of dissolving the recovered precipitate with 0.2N saline and then performing ethanol precipitation was repeated. This SDS-treated precipitate is
By subjecting the gel to DS gel electrophoresis, a portion that did not penetrate into the gel was recovered. After dialyzing the collected fraction, DEA
Purified polysaccharide was obtained as an adsorbed fraction by ion-exchange chromatography using 650 M of E-Toyopearl (manufactured by Tosoh Corporation) and freeze-dried to obtain a phosphorylated polysaccharide of the formula (I) having m of 0 to 3 and n of 1,000 to 5,000. About 700 mg of saccharide powder was obtained.

[0013]

Example 1 Each substance was mixed and dissolved according to the formulation shown in Table 1 below.

[0014]

[Table 1] 70% sorbitol 3.0 (g) glycerin 5.0 Phosphorylated polysaccharide obtained in Reference Example 1 1.0 Water 70.0 ────────────────────────

To this solution, 0.1 g of allantoin, 0.5 g of polyoxyethylene hydrogenated castor oil derivative, and 20.
0 g and a little perfume were added with stirring to form a uniform solution to produce an oily lotion having the effects of preventing and improving skin aging.

[0016]

Example 2 Each substance was mixed and dissolved according to the formulation shown in Table 2 below.

[0017]

[Table 2] Ethylene glycol 2.5 (g) Propylene glycol 5.0 Silicone 0.0001 Phosphorylated polysaccharide obtained in Reference Example 1 1.0 Water 70.0 ────────────────────────

To this solution, 1.0 g of urea, 1.2 g of polyoxyethylene sorbitan monolaurate, 20.0 g of ethanol
g, 0.1 g of ε-aminocaproic acid and a little fragrance were added with stirring to form a uniform solution, thereby producing a dry lotion imparting the effect of preventing and improving skin aging.

[0019]

Example 3 Each substance was mixed in the composition shown in Table 3 below and dissolved by heating at 75 ° C.

[0020]

[Table 3] ───────────────────────── Glycerin 2.0 (g) Stearic acid 20.0 Myristic acid 10.0 Lauric acid 5.0 Polyoxyethylene lauryl ether 1.0 Reference Phosphorylated polysaccharide obtained in Example 1 1.0 Fragrance a little Preservative a little ─────────────────────────

Into this solution, 5.5 g of potassium hydroxide and water
After adding 56.0 g with stirring, the mixture was cooled to produce a facial cleansing cream having the effects of preventing and improving skin aging.

[0022]

Example 4 Each substance was mixed in the composition shown in Table 4 below and dissolved by heating at 75 ° C.

[0023]

[Table 4] ────────────────────────────── Beeswax 10.0 (g) Paraffin wax 6.0 Lanolin 3.0 Isopropyl myristate 6.0 Squalane 8.0 Liquid paraffin 25.0 Phosphorylated polysaccharide obtained in Reference Example 1 1.0 Polyoxyethylene sorbitan monostearate 1.8 Sorbitan monostearate 4.2 Preservative a little ─────────────────── ───────────

In this solution, propylene glycol 2.0 was added.
g, borax 0.7 g, urea 5.0 g and water 28.0 g were added with stirring, then cooled, and a small amount of fragrance was added to produce an oily cream having an effect of preventing and improving skin aging.

[0025]

Comparative Example 1 As a comparative example, an oil-based cream was prepared in the same manner as in Example 4 except that sodium hyaluronate was used instead of the phosphorylated polysaccharide.

[0026]

[Test Example 1] For each of 20 women aged 20 to 50 years, the oil cream of Example 4 or the oil cream of Comparative Example 1 was used twice a day and twice a day for one month.
Changes in skin moisturizing, softness and elasticity were examined. With respect to the results, Table 5 shows changes in skin moisture retention, Table 6 shows changes in skin flexibility, and Table 7 shows changes in skin elasticity.

With respect to the moisturizing property of the skin, Transepidermal Water Loss (Transepidermal Water Loss) indicating the amount of water that is insensitively lost from the skin after using each oily cream.
s: TWL) value was measured with an evaporometer and evaluated according to the following criteria. Very effective: TWL value decreased by 30% or more compared to the value of Comparative Example 1. Slightly effective: TWL value was 10% or more compared to the value of Comparative Example 1.
Less than 30% reduction No effect: TWL value decreased by less than 10% or increased by less than 10% compared to the value of Comparative Example 1 Deterioration: TWL value increased by 10% or more compared to the value of Comparative Example 1 Also, change in skin flexibility And the change in skin elasticity was represented by sensory evaluation of the panel.

[0028]

[Table 5] ─────────────────────────────── (very effective) (somewhat effective) (no effect) (Aggravated) ─────────────────────────────── 4/20 12/20 4/20 0/20 ──── ───────────────────────────

[0029]

[Table 6] ─────────────────────────────────── (very improved) (slightly improved) (change) None) (worse) ─────────────────────────────────── Example 11/20 9/200 / 20 0/20 Comparative Example 9/20 11/20 0/20 0/20 ─────────────────────────────── ────

[0030]

[Table 7] ─────────────────────────────────── (very improved) (slightly improved) (change) None) (worse) 実 施 Example 9/20 10/20 1 / 20 0/20 Comparative Example 7/20 12/20 1/20 0/20 ─────────────────────────────── ────

According to the results of this test, the phosphorylated polysaccharide has an excellent moisturizing property and has an effect of improving skin softness and elasticity.

[0032]

Test Example 2 In this test, the activity of the phosphorylated polysaccharide obtained in Reference Example 1 for promoting collagen production was examined. Regarding the activity of promoting collagen production, the C-terminal pro-collagen released by the action of endopeptidase when collagen is biosynthesized as procollagen in cells and this procollagen is secreted extracellularly and polymerized as collagen fibers. Peptide (PIP) was determined by measuring by ELISA. In this test, human normal fibroblasts (IMR-90), for which collagen-producing ability was recognized, were used.

That is, IMR-90 (ATCC CCL-186) cells were
After culturing in a well plate until confluence, the cells were washed with PBS. Next, a phosphorylated polysaccharide solution prepared in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum (FCS) to a final concentration of 1%, and this phosphorylated polysaccharide solution as a stock solution was used as a stock solution. A 2-fold serially diluted phosphorylated polysaccharide solution was added to each well, and after further culturing for 3 days, PIP contained in the supernatant was added.
The amount was measured with an ELISA kit (Takara Shuzo). The result is shown in FIG.

According to the results of this test, phosphorylated polysaccharide promotes collagen production in a concentration-dependent manner. That is, by adding 0.1%, 0.5%, or 1% of the phosphorylated polysaccharide, the collagen production promoting activity is 1.1 times, 1.4 times, as compared with the case where no phosphorylated polysaccharide is added.
Or you can see that they increase by 1.7 times respectively.

[0035]

The effect of enhancing skin function can be imparted by blending a skin aging preventive and improving agent containing a phosphorylated polysaccharide, which is produced by lactic acid bacteria, which is extremely safe, as an active ingredient into cosmetics and the like. , Can prevent and improve skin hardening, tension, wrinkles and the like.

[Brief description of the drawings]

FIG. 1 shows the collagen production promoting effect of phosphorylated polysaccharide in Test Example 2.

 ──────────────────────────────────────────────────続 き Continuation of the front page (72) Inventor Yukiyuki Ota Hokkaido, Sapporo City, Atsetsu-ku, Atsubetsu Chuo 5-chome 9-9-10-306 (72) Inventor Klaas Dusburg, Netherlands, 9602 DOUBLEZY, HOGEZAND, Fan Hohendorpstraat 2 (72) Inventor Jan Sickema, The Netherlands, 9483 P.A. Zeigs, Hofdebege 1B (72) Inventor, Taisuke Iwasaki, Netherlands 9602 HK Hogezand, Princess Larn 4

Claims (4)

[Claims] 1. An agent for preventing and improving skin aging comprising a phosphorylated polysaccharide as an active ingredient. 2. The agent for preventing and improving skin aging according to claim 1, wherein a phosphorylated polysaccharide represented by the formula (I) is used. Embedded image (Where Glc represents a glucose residue, Gal represents a galactose residue, and Rha represents a rhamnose residue, respectively.
In addition, the numbers written in large letters in the formula represent each binding site, m is an integer of 0 to 3, n is a repeating unit,
Shows 1,000 to 5,000, respectively. ) 3. The preventive and ameliorating agent for skin aging according to claim 1, wherein the phosphorylated polysaccharide represented by the formula (II) is used. Embedded image (Where Glc represents a glucose residue, Gal represents a galactose residue, and Rha represents a rhamnose residue, respectively.
In the formulas, the numbers written in large letters indicate the respective binding sites, and n indicates the unit of repetition, and indicates 1,000 to 5,000, respectively. ) 4. The agent for preventing and improving skin aging according to claim 1, wherein the agent for preventing and improving skin aging is a cosmetic.