JP2557240B2 - Skin anti-aging agent - Google Patents

Description

TECHNICAL FIELD The present invention relates to a skin aging preventive agent (used for preventing skin aging). Specifically, skin aging prevention effect (rough skin improvement effect, keratin improvement effect, effect of increasing skin turnover speed, effect of improving skin dullness, sagging, acupuncture, gloss, hardening, skin activating effect, moisturizing effect, etc. ).

[Conventional technology]

Regarding the aging phenomenon of skin with aging (a phenomenon often seen in old people), generally, it becomes macroscopically dull, wrinkles, and spots and becomes a dry state, and along with it, so-called skin moisturization, gloss, smoothness, It is known to cause functional changes such as reduction in softness and elasticity. In addition, skin diseases that are frequently observed in the elderly (illnesses that are common in the elderly) include senile pigmented spots, senile xerosis, senile vitiligo, senile warts, senile neurofibroma, and senile faces. Examples include blisters and senile sebaceous hyperplasia. Further, it is known that malignant tumors such as cancer precursor (keratokeratosis, cutaneous horn, Bowen's disease, malignant melanoma), skin cancer, malignant melanoma, etc. occur.

Factors of these aging phenomena are internal factors such as sebaceous gland hypofunction, sweat gland hypofunction, rough stratum corneum, epidermal thinning, increased collagen cross-linking, increased insoluble collagen, decreased connective tissue matrix, decreased capillary function, External factors such as a decrease in metabolic function and abnormal pigment metabolism include ultraviolet rays, low temperature and low humidity conditions.

Dry and non-smooth rough skin, which is a typical feature of aging skin, has a decrease in keratinocyte function, which leads to a decrease in sebum secretion due to aging and a quantitative decrease in water-binding substances in the epidermis (especially in the stratum corneum). Is a symptom induced by lack of water. In the elderly, the stratum corneum water content tends to occur when the water content is about 10% or less of the saturated water content. This symptom is exacerbated particularly when the air is dry in the winter, and preferable examples include senile xerosis.

In order to deal with such a dry skin phenomenon, skin turnover (the speed at which the epidermis changes to the stratum corneum and peels off: since aging skin has a slow cell division rate) It is necessary to maintain normal skin function while increasing turnover (delay), and various drugs have been developed in the medical and cosmetic fields. As a general method, there are a vasodilator such as a vitamin E agent and a respiratory stimulant such as placenta liquid, which promotes improvement by promoting skin fine structure and increasing metabolism. There is also a method of applying a composition similar to sebum or a moisturizing agent to the surface of the skin to cover the skin to prevent the skin from drying.

[Problems to be solved by the invention]

However, the ameliorating effect due to the above-mentioned vasodilatory action or skin respiration promoting action is slow-acting, and it takes at least one week for a cream and at least two weeks for a lotion until the effect is exhibited, which is quite satisfactory. There was room for improvement because I couldn't get it. Also, sebum-like compositions and moisturizers temporarily help retain water in the stratum corneum, but their usability is poor and they are greasy or greasy. There was a drawback that the (coating amount) had to be increased. Further, as a drug for coping with aging phenomena other than those described above (for example, wrinkles, sagging, and stains), a drug which is still satisfactory in terms of efficacy and safety has been obtained in the medical and cosmetic fields. Absent.

In view of the above circumstances, the present inventors have conducted extensive studies to obtain a drug that is effective and highly safe against aging, and as a result, sialic acid and sialic acid derivatives have excellent anti-aging effect. The present invention has been completed and the present invention has been completed.

[Means for Solving the Problems] That is, the present invention provides a skin aging preventive agent containing one or more selected from the group consisting of sialic acid and sialic acid derivatives.

 Hereinafter, the configuration of the present invention will be described in detail.

Sialic acid used in the present invention is a known substance, and is a generic term for N-acyl and N-acyl, O-acyl derivatives of polyhydroxymonoaminocarboxylic acid (inolaminic acid), and is also called sialic acid. Sialic acid is contained in various organisms, and normally constitutes a cell surface saccharide and is present in a glycoside bond with the ends of sugar proteins and glycolipids. In particular, it is mainly present in the brain, nerves, blood, submandibular gland, mucinous substance, and colostrum, but is also liberated and is also present in serum, body fluids, and urine. The most common sialic acid found in the living world is N-acetylneuraminic acid (NANA: molecular weight 309), melting point 185-187 °,
▲ [α] ²⁰ _D ▼ -32 ° ± 2 ° white powder.

The biological pharmacological actions of sialic acid and sialic acid derivatives are known to contribute to the negative charge on the cell surface, involve in the specific recognition mechanism of cells, hemagglutination reaction by viruses, homing phenomenon of blood proteins, etc. ing. Recently, with regard to sialic acid, expectorant activity (JP-A-61-6841
8), anti-inflammatory effect (JP-A-62-145015) and nutritional effect of sialic acid-linked oligosaccharide (JP-A-61-152233).

However, it is not known to have a skin aging preventive effect. Examples of the production method of sialic acid and sialic acid derivatives include synthetic methods and natural extraction methods (raw materials: egg white, submandibular gland mucin, placenta extract, colominic acid obtained from Escherichia coli, etc.). Law (Patent No. 119121
2, the application of Japanese Patent No. 1253988, JP-A-59-184197, etc.) is particularly preferable.

The production examples of the compound according to the present invention with a milk extract are shown below. Milk using the milk extraction method can be appropriately selected from cow milk, sheep milk, breast milk (human) and the like. Further, since the sialic acid content concentration can be freely controlled depending on the degree of purification, it may be purified according to the usage, application, dosage form and other purposes. The manufacturing method is not limited to this.

(Production Example 1) Sialic acid 10 g of cheese whey was added with 28 kg of hydrochloric acid to adjust the pH to 2, and then heated at 92 ° C. for 60 minutes for hydrolysis. The obtained whey hydrolyzate was heated with a screw decanter to remove proteins coagulated by heating to obtain a sialic acid-containing liquid. Next, the sialic acid-containing liquid was placed in the desalting tank of the electrodialyzer to desalinize the acid to a conductivity of 2 mS / cm. Then add 30 distilled water to the concentration tank.
Then, the solution was dialyzed again to a conductivity of 50 μS / cm. After the completion of dialysis, the concentration tank liquid was concentrated and dried to obtain 600 g of a white powder. The free sialic acid purity of the powder thus obtained was about 97%.

(Production Example 2) sialic acid salt The above-mentioned free sialic acid was adjusted with caustic soda to prepare sialic acid.
Get Na. Besides, Mg salt, k salt, phosphate, amine salt and the like can be produced according to the purpose.

(Production Example 3) Sialic acid-bound oligosaccharide The filtrate prepared by ultrafiltration of milk or cheese whey was concentrated under reduced pressure to a total solid content of 20%. 25 kg of this solution was electrodialyzed until the electric conductivity reached 20 μS / cm, and then passed through a column packed with Dowex 1 × 2 (2 kg) to adsorb oligosaccharides. Next, 20 kg of water was passed through this column to remove the neutral sugar, and then 0.5 M hydrochloric acid was passed through to elute the oligosaccharide adsorbed on the column. The pH of the obtained eluate was adjusted to pH 7.0 with 30% caustic soda, and electrodialysis was performed again until the electric conductivity reached 150 μS / cm. The obtained dialysate was concentrated and dried to obtain 100 g of white powder. The powder thus obtained had a sialic acid-linked oligosaccharide purity of 90%.

(Production Example 4) Sialic acid-bound protein Rennet gazein curd waste liquid 300 was sterilized by heating in a Fordora tank at 75 ° C for 30 minutes, then cooled to 40 ° C, and the pH was adjusted to 4.6 with concentrated hydrochloric acid. After allowing the solution to stand for about 30 minutes to form a precipitate and passing the solution through a clarifier to return the pH to 7.0, the clear solution was concentrated and desalted with an RO membrane. After concentrating to 10 times, it was added to the concentrated solution and concentrated again to 10 times. The purity of the obtained powder of sialic acid-bound protein was about 80%.

(Production Example 5) Chloroform-methanol mixture (1: 1) was added 100 to 15 kg of buttermilk powder of sialic acid lipid, and the mixture was stirred at room temperature for 30 minutes for extraction. Next, the chloroform-methanol mixed solution was collected and passed through an ion exchange resin (DEAE-Toyopearl acetate type) 1 to adsorb the sialic acid-bound lipid, and then the resin was washed with the chloroform-methanol mixed solution (1: 1) 2 and then 0.5 After washing the resin with M sodium acetate-methanol solution (1: 1) 2, the sialic acid-bound lipid was eluted with 0.5 M sodium acetate-methanol solution 4. The eluate was concentrated to remove methanol, 5 g of water was added, and the mixture was desalted by ultrafiltration and the sialic acid-bound lipid was concentrated. After the concentrated solution containing sialic acid-bonded lipids was evaporated to dryness with an evaporator, a silica gel column (4.0
X50 cm) to adsorb the sialic acid-bound lipid, and then chloroform 1, chloroform-methanol mixture (4:
1) After sequentially washing with 1, the sialic acid-bound lipid was eluted with a mixed solution 2 of chloroform-methanol (2: 8). The eluate was evaporated to dryness with an evaporator to obtain 15 g of a white powder.

The purity of the sialic acid-bound lipid in the powder thus obtained was 90% or more by thin layer chromatography (resorcinol method).

Next, the safety test of sialic acid and sialic acid derivatives was carried out, and the results are shown.

Acute toxicity test Each of the sialic acid, Na sialic acid, sialic acid-binding oligosaccharide, sialic acid-binding protein and sialic acid-binding lipid produced in Production Examples 1 to 5 was used as a test substance, and ICR female mice (6 weeks old, 1 For intraperitoneal administration, a suspension of 0.5% carboxymethylcellulose sodium / physiological saline solution to 2% by weight of each test substance was prepared using a 22 1/2 G injection needle. Oral administration was performed by intraperitoneally administering (60 ml / kg) or oral administration (30 ml / kg) by suspending 5% by weight of each test substance in olive oil using an oral needle. As a result, none of the test substances was found dead during the 7-day observation period, decreased in body weight, or toxicosis that should be noted, and none was observed in the autopsy performed on the 7th day. .

As is clear from this result, the acute toxicity value (LD ₅₀ ) of sialic acid and sialic acid derivative was 1200 mg after intraperitoneal administration.
/ kg or more and 1500 mg / kg or more by oral administration, it was proved to be highly safe.

Next, formulation for applying sialic acid and sialic acid derivatives as a skin aging preventive agent will be described.

The skin anti-aging agent of the present invention can be used in the form of a mixture containing a pharmaceutically acceptable additive and other drug in addition to sialic acid and a sialic acid derivative. However, since sialic acid has poor stability to light, acid and alkali, it has a problem of long-term storage stability, and it is preferable to use a sialic acid derivative from the viewpoint of stability.

In the skin aging preventive agent of the present invention, in addition to the above-mentioned essential components, other components usually used in cosmetics and pharmaceuticals, such as avocado oil, palm oil, peanut oil, beef tallow, rice bran oil, jojoba oil, carnauba wax, lanolin, Oils such as liquid paraffin, oxystearic acid, isostearyl palmitate, isostearyl alcohol, glycerin,
Sorbitol, polyethylene glycol, collagen,
Moisturizers such as hyaluronic acid and chondroitin sulfate, UV absorbers such as amyl paradimethylaminobenzoate, urocanic acid and ethyl diisopropylcinnamate, antioxidants such as sodium erythorbate, sage extract, vitamin E and parahydroxyanisole, stearyl Sodium sulfate, diethanolamine cetylsulfate, cetyltrimethylammonium saccharin, polyethylene glycol isostearate, surfactants such as glyceryl arachiate, preservatives such as ethylparaben, butylparaben, etc. Extract, glycyrrhizic acid derivative, glycyrrhetinic acid derivative, salicylic acid derivative, hinokitiol, zinc oxide, anti-inflammatory agents such as allantoin, placenta extract, glutathione, snow Sita extracts, whitening agents such as ascorbic acid derivatives, carrot, aloe, mallow, iris, grape, Coix seed, dishcloth gourd extract, such as lily, royal jelly, photosensitizer, cholesterol derivatives,
Various amino acids, biotin, activators of various vitamins and derivatives such as pantothenic acid derivatives, extracts of saffron, senkyu, ginger, Hypericum, ononis, rosemary, garlic, γ-oryzanol, dextran sulfate sodium, vitamin E Derivatives, nicotinic acid derivatives, blood circulation promoters such as nicotinic acid derivatives, antiseborrheic agents such as sulfur and thiantol, fragrances, water, alcohols, thickeners such as carboxyvinyl polymers, titanium yellow, cursamine, safflower red, etc. Agents and the like can be appropriately blended as necessary.

If the dosage form of the present invention is suitable for obtaining a medicinal effect,
It can be used in any form, and examples thereof include agents, powders, granules, powders, tablets, capsules, syrups, suppositories, injections and the like. Of these, external preparations are convenient in terms of use and preferred for both prevention and treatment, and may be in any form as long as they can be applied externally.

For example, lotions, liniments, external liquids such as emulsions,
Examples thereof include external semi-solid preparations such as cream, ointment, pasta, jelly, spray, etc., and the administration method is percutaneous administration, which is directly applied to or sprayed on the skin.

In the agent for preventing skin aging of the present invention, the compounding amount of one or more of sialic acid and sialic acid derivative as an active ingredient is 0.0001 to 50% by weight, preferably 0.001 to 5% by weight based on the total weight of the composition. %. If it is less than 0.0001% by weight, the effect tends to be poor, and even if it exceeds 50% by weight, it is difficult to expect a great increase in the effect.

Dosage of the skin aging preventive agent of the present invention, age, individual differences,
It is possible to pass an amount outside the following range depending on the symptoms etc., so it is not possible to specify clearly, but in general, one or more sialic acid and sialic acid derivatives are orally administered and the transdermal dose is 1 kg body weight and 1 day. The amount is 0.0001 to 2000 mg, preferably 0.001 to 200 mg, and this amount can be administered once a day or in several divided doses.

Further, depending on the concentration of sialic acid and sialic acid derivative and the desired degree of effect, it can be applied as a food for the purpose of preventing drugs such as pharmaceuticals, quasi drugs, cosmetics and aging.

[Examples] In order to clarify the excellent effects of the present invention, evaluation tests were performed using Examples and Comparative Examples. The present invention is not limited to this.

(Test method) (1) Method of measuring turnover speed of epidermis using guinea pig skin Albino guinea pigs were divided into 6 groups, each group consisting of 5 animals, and used for evaluation. After shaving the abdominal skin of the guinea pig, white vaseline ointment containing 5% by weight of the fluorescent dye dansyl chloride was applied occlusion for 24 hours at 0.1 ml / cm ² . As test substances, 0.3 ml of each of Examples 1 to 5 and Comparative Example 1 was applied once a day on the dansyl chloride coating part, and the stratum corneum turnover speed was evaluated by the number of days until the disappearance of dansyl chloride.

(2) Method for measuring the therapeutic effect on xeroderma disease 30 female senile xerosis patients aged 40 to 60 were used as subjects,
Examples 1 to 5 in which 5 people in 1 group are divided into 6 groups and a drug is mixed in the right cheek
Alternatively, Comparative Example 1 was applied, and Comparative Example 2 containing no drug was applied as a control to the left cheek. The test period was one month in winter, and 2 ml each of the test substance was applied twice a day in the morning and evening. At the end of application for 30 days, the doctor determined the improvement effect on the right cheek compared to the left cheek.

(3) Method for measuring activating effect on tired human skin Due to lack of sleep, skin roughness, darkening of the skin, gloss of the skin, disappearance of acupuncture, etc. occur. In particular, in the winter when these symptoms remarkably occur, the immediate skin activating effect of the test substance on the tired skin was evaluated. Thirty women in their thirties were used as test subjects, and each group was divided into 6 groups, 5 groups, and the right cheek was coated with Examples 1 to 5 or Comparative Example 1 and the left cheek was uncoated. The test period is 2 days in winter, and 2 ml each of the test substance is used for 2 days a day.
It was applied twice in the morning and before going to bed, and after the test was completed, the subject self-scored the skin condition after the test as compared with the skin before the test.

However, during the two days of the test, the test subject shall stay in the bed at 4 am at 7 am in the morning (sleep time 3 hours) and take no other nap.

-Evaluation items-Roughness of skin, dullness of skin, texture of skin, paste of makeup, gloss of skin, swelling, and sagging-Score-5 points: markedly improved compared to before the test.

4 points: 〃 Some improvement.

3 points: 〃 No change.

2 points: 〃 Somewhat worse.

1 point: 〃 Significantly worsened.

(4) Method of measuring aging preventive effect by human skin Using 30 women aged 25 to 50 as test subjects, one group is divided into 6 groups with 5 persons, and Examples 1 to 5 and Comparative Example 1 are used on the right cheek, left side. Comparative example 2 containing no drug was used on the cheek. The test period was two months in winter, and continuous application was performed twice a day in the morning and night, and the effects were evaluated by the following (a) to (e).

(A) Evaluation of skin surface morphology The skin surface morphology was evaluated by visually determining the skin dryness according to the following criteria.

-Judgment method- Remarkable effect: Compared to Comparative Example 2, there is almost no drying or dropping.

Effective: Compared to Comparative Example 2, there is less drying and falling.

Slightly effective: Drying is similar to that of Comparative Example 2, but there is no drop.

Ineffective: There is no difference in drying and dropping compared with Comparative Example 2.

Deterioration: Drying and dropping are remarkable as compared with Comparative Example 2.

(B) Observation of Skin Appearance The appearance was evaluated by collecting skin replicas of the above subjects using a silicone resin and observing them with a stereomicroscope.

-Judgment method- Remarkable effect: The skin crevices and cuticles are remarkably clear and arranged as compared with Comparative Example 2.

Effective: Compared with Comparative Example 2, the sulcus and the cuticle are slightly clearer.

Invalid: No difference is recognized as compared with Comparative Example 2.

Deterioration: As compared with Comparative Example 2, the skin groove and the cuff are less clear.

(C) Evaluation of stratum corneum function The function of the stratum corneum was evaluated by measuring TWL (Trans-epidermal Water Loss).
The TUL was measured using an evaporation meter EPL (Sweden servo Med.).

-Judgment method- Remarkable effect: TWL is reduced by 30% or more as compared with Comparative Example 2.

Effective: TWL is reduced by 10% or more compared to Comparative Example 2.

Invalid: TWL is less than 10% compared to Comparative Example 2.

Deterioration: TWL increased by 10% or more as compared with Comparative Example 2.

(D) Evaluation of exfoliated keratinocytes After adhering Scotch tape (Nichiban Mending Tape) to the skin of the test site before and after the evaluation test and exfoliating it,
The keratinocytes on the tape were stained with HE and the presence or absence of nuclei in the cells was determined by a microscope.

-Judgment method-Normal: No nucleus is observed at all and normal keratinization occurs.

Above: Nucleus is observed and abnormal keratinization occurs.

(E) Sensory Evaluation After the test was completed, the subject self-scored the skin condition of the right cheek as compared to the left cheek (Comparative Example 2).

-Evaluation items-Wrinkles, texture of the skin, dull skin, sagging skin, gloss, balm, and makeup paste-Score-5 points: markedly improved compared to Comparative Example 2.

4 points: 〃 Some improvement.

3 points: 〃 No change.

2 points: 〃 Somewhat worse.

1 point: 〃 Significantly worsened.

Comparative Example 2 is a formulation in which the vitamin E acetate of Comparative Example 1 is replaced with purified water and no drug is mixed.

(Test Results) (1) As is apparent from the results in Table 2, Examples 1 to 5 containing sialic acid or a sialic acid derivative were compared with Comparative Example 1 by the evaluation using guinea pigs, and the turnover of the epidermis was higher. It was confirmed that the temperature was accelerated for 2 to 3 days, and it was found that a synergistic effect can be obtained by combining two or more kinds.

(2) From the results of Table-3, it was confirmed that Examples 1 to 5 were more effective in senile xerosis than Comparative Example 1. Furthermore, it was also found that a synergistic effect can be obtained by combining two or more kinds of sialic acid and sialic acid derivatives.

(3) From the results of Table-4, Examples 1 to 5 are Comparative Example 1
It was found that compared with the above, there was an immediate skin activating effect on tired skin, and a synergistic effect was obtained by combining two or more kinds of sialic acid and sialic acid derivatives.

(4) From the results of Table-5, macroscopic, replica, TWL,
In all the evaluations of sensory properties, it was confirmed that the examples were more effective than the comparative example 1 and that the combination of sialic acid and the sialic acid derivative had a synergistic effect. (D)
The results show that the effect of promoting epidermal turnover is not caused by abnormal keratinization.

Next, various embodiments will be shown. In the following, the blending amount will be% by weight.

Example 6 Emulsion An emulsion was prepared by a conventional method according to the following formulation.

Stearic acid 2.0 (%) Cetanol 1.0 Vaseline 3.0 Lanolin alcohol 2.0 Liquid paraffin 8.0 Squalane 3.0 Escalol 507 2.0 Sodium sialate 0.001 POE (10) Monooleate 2.5 Triethanolamine 1.0 Propylene glycol 5.0 Preservatives appropriate amount Fragrance appropriate amount Distilled water Residual example 7 Nutrition cream A nutrition cream was produced by a conventional method according to the following formulation.

Stearic Acid 2.0 (%) Stearic Alcohol 7.0 Reduced Lanolin 2.0 Squalane 5.0 Octyldodecanol 6.0 POE (25) Cetyl Ether 3.0 Glyceryl Monostearate 2.0 Propylene Glycol 5.0 Sialic Acid-Binding Oligosaccharide 1.0 Sialic Acid-Binding Protein 1.0 Preservative Appropriate Fragrance Appropriate amount Distilled water Residue Example 8 Pack A pack was produced by a conventional method according to the following formulation.

Kaolin 67.5 (%) Talc 19.0 Propylene glycol 7.5 Calcium acetate 0.01 Urine 0.5 Sialic acid-bound lipid 5.0 Fragrance 0.49 Example 9 Lip Treatment A lip treatment having the following formulation was prepared according to a conventional method.

Candelilla wax 9.0 (%) Solid paraffin 8.0 Beeswax 5.0 Carnauba wax 5.0 Lanolin 11.0 Sialic acid-binding oligosaccharides 0.2 Isopropyl myristate 10.0 Fragrance Suitable amount Antioxidant Suitable amount Castor oil Residue Example 10 Ointment The ointment prepared according to the conventional method did.

Stearyl alcohol 18.0 Mokurou 20.0 Sialic acid-binding protein 0.5 Polyoxyethylene molooleate 0.25 Glycerin monostearate 0.25 Vaseline 4.0 Purified water Residue [Effect of the invention] The skin aging preventive agent of the present invention is effective for preventing skin aging (rough skin). Improvement, keratin improvement, epidermal turnover acceleration, skin dullness, sagging, acupuncture, gloss, hardening improvement, skin activation hardening,
It has excellent moisturizing effect), stability, safety and usability.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Yukio Togawa 157 Idogaya Yanaka-cho, Minami-ku, Yokohama-shi, Kanagawa 703 Diamond Palace Idogaya 703 (72) Inventor Kenkichi Ahiko 2-688-18 Hanakoganei, Kodaira-shi, Tokyo ( 72) Inventor Eiji Eike 2-23-5 Irumagawa, Sayama-shi, Saitama 102 (56) Reference JP-A-62-145015 (JP, A)

Claims (4)

(57) [Claims] 1. A skin aging preventive agent containing one or more selected from the group consisting of sialic acid and sialic acid derivatives. 2. A sialic acid derivative is a sialic acid-linked oligosaccharide,
The agent for preventing skin aging according to claim (1), which is a sialic acid-binding protein, a sialic acid-binding lipid or a sialic acid salt. 3. The skin aging preventive agent according to claim (1) or (2), which uses a sialic acid or a sialic acid derivative obtained by extraction from milk. 4. The sialic acid is N-acetylneuraminic acid. Claims (1), (2) or (3).
The agent for preventing skin aging according to the item.