JP4926448B2 - Anti-aging agent - Google Patents

Description

The present invention relates to an anti-aging agent characterized in that it contains an active ingredient extracted from the brown algae Sargassum horneri with water , and more specifically , cellulase in the extraction treatment step. The present invention relates to an anti-aging preparation having excellent hyaluronidase activity-suppressing action and superior stability over time and alcohol stability.

In recent years, health consciousness has been deeply rooted, and efforts have been made to maintain an internally healthy body and spirit while paying attention to eating habits. For example, active intake of health foods and functional foods has emerged. In addition, there are many variations of supplementary foods, so-called supplements are a new form of food that is completely established in general consumers.

Under such circumstances, multi-functionality such as immunostimulatory action of materials such as algae has attracted attention, and their application development is particularly active.

On the other hand, not only the form to eat in this way but the development which intended skin care is also tried, and the application example to the cosmetics of brown algae is also well-known.
JP-A-2-124810 JP 7-10769 A Japanese Patent Laid-Open No. 10-236918 For example, Japanese Patent Laid-Open No. 2-124810 discloses that an extract of seaweeds belonging to the order Hibamataceae and Kombu is useful as a cosmetic having a whitening effect and an astringent effect. . JP-A-7-10769 discloses a method for controlling the astringency of seaweeds, and JP-A-10-236918 discloses a patch for ensuring the moisture retention of seaweeds. Further, Japanese Patent Application Laid-Open No. 10-330219 discloses a seaweed whitening agent containing akamoku (Sargassum horneri).

However, the greatest difficulty in blending the functional raw materials related to the seaweed into the skin care preparation is the restriction on the preparation design due to odor and stability.

In other words, the seaweed extract contains a lot of water-soluble polysaccharides such as alginic acid, so it is excellent in skin familiarity and moist feeling, but has a habit odor peculiar to seaweed and gives consumers unpleasant feeling during application. It was. In addition, the raw material itself had poor temperature resistance, and a strange odor was generated due to deterioration over time. Furthermore, especially in lotion-based preparations containing a large amount of alcohol, the raw materials cannot coexist with alcohol for a long time, causing precipitation and sediment, which is a major limitation in the formulation design. In addition to this, there has been a problem of a decrease in safety due to the incorporation of a large amount of stabilizers and preservatives.

On the other hand, the functionality required of cosmetics is strongly demanded to be effective in preventing aging in order to satisfy the fundamental human desire to remain young and beautiful forever.

In order to solve such problems, the present inventor considered to further apply and develop a seaweed extraction technique.
As a result of various studies, a desired effect can be found particularly in Sargassum horneri, and the present invention has been completed.

An object of the present invention is to find a raw material that allows consumers to realize an anti-aging effect without feeling unpleasant odor, and to provide an anti-aging agent containing the raw material.

As a result of continual search to solve the above-mentioned conventional problems, the present inventor suppresses hyaluronidase activity excellent in stability by performing a specific extraction process on Sargassum horneri. As a result, the present invention has been completed.

That is, the anti-aging agent of the present invention is characterized in that the extract of Sargassum horneri is extracted with water, and the active ingredient is appropriately purified using cellulase in the extraction process. is there.

ADVANTAGE OF THE INVENTION According to this invention, the anti-aging agent which has a function which prevents a wrinkle, dullness, etc. of skin by the remarkable inhibitory action of hyaluronidase activity and normalizes a skin state is provided.

Hereinafter, embodiments of the present invention will be described in detail.
The anti-aging agent of the present invention is characterized in that it contains an active ingredient extracted from the genus Sargassum horneri identified from among the seaweeds of the family A. Here, Akamoku is a kind of brown algae that has been used mainly as an edible seaweed for a long time in Japan.

The term “Sargassum horneri” extract of the present invention refers to an extract obtained by extracting red sardine (Sargassum horneri) with water under certain conditions. Below, the suitable manufacturing method of the akamoku extract which is an active ingredient of this invention is described.

Usually, water is added to red sardine (Sargassum horneri) at a ratio of 1: 2 to 1: 100, and the mixture is extracted by stirring for 1 to 48 hours at a temperature of 5 to 90 ° C. A lower alcohol such as ethanol is usually added 1 to 5 times the weight of the filtrate obtained by purifying the extract through a purification process such as filtration or centrifugation, but there is a marked change in stability improvement even if it is added 2 times or more. Since it is not seen, it is preferable to add 1 to 2 times the amount in consideration of safety and cost. By purifying the obtained precipitate after dissolving it in water, the akamoku extract of the present invention (pH 4.0 to 7.0) can be obtained.

In general, it is preferable to cut the starting raw material akamoku (scientific name: Sargassum horneri) without drying it in consideration of the loss of the extracted components.

During extraction, it is preferable to adjust the pH to 3.0 to 6.0 in terms of improving the stability of the product. When extraction is performed at a pH of 6.0 or more, the stability of the product is remarkably lowered, and the product value as a cosmetic raw material is lowered because the coloring is remarkable. On the other hand, when the pH is less than 3.0, problems such as a decrease in extraction efficiency and decomposition of active ingredients may occur.

In order to increase the extraction efficiency of the active ingredient, it is preferable to heat and stir, but in order to avoid the extraction of the brown substance that is a destabilizing factor of the extract, it is usually set to 5-90 ° C, preferably 40-60 ° C. To do.

In this extraction treatment step, an enzyme treatment is performed to further improve the extraction efficiency of the active ingredient. Cellulase can be preferably used as the enzyme. As a precaution for addition, adjustment to the optimum pH of cellulase can be mentioned, and it is usually set to 4.0 to 5.5. After inactivating the enzyme by heat treatment at least at 90 ° C. for 30 minutes, a filtrate is obtained.

A lower alcohol such as methanol, ethanol, propanol or butanol, preferably ethanol, is added to the filtrate in an amount of 1 to 2 times the weight of 1 part of the filtrate to dissolve and remove impurities that adversely affect the quality. At the time of removal, it is preferably centrifuged at 5,000 to 10,000 rpm for 15 minutes to 60 minutes, but after that, by adding water to the separated precipitate and refiltering, A red mock extract is obtained.

The extract of the present invention thus obtained may be used as it is, but is usually used by blending it with a preparation. The form of the preparation can be provided for external use, but a base generally acceptable for cosmetics is selected and applied directly to the affected area. In this case, in addition to homogeneous preparations represented by lotions and essences, general emulsification systems such as O / W and W / O types represented by creams and emulsions, W / O / W, O / W / O-type special multi-layer emulsions, other types such as pastes, ointments and tinctures, sprays such as aerosols and sprays, patch types such as poultices and plasters, etc. The basic base is also used in combination with other components and is not particularly limited.

In these present inventions, the amount of akamoku extract blended is in the case of cosmetics such as creams, lotions, emulsions, packs, lotions, essences, etc. and when used as a dosage form such as sheet masks, poultices, plasters, etc. In any case, it is blended in the range of 0.1 to 50% by weight, preferably 0.1 to 10% by weight, based on the whole preparation. When the amount is less than 0.1% by weight, the anti-inflammatory action and the active oxygen scavenging action are insufficient. Moreover, even if it exceeds 50% by weight, there is no particular difference in effect from the case of less than that, and there is a problem that this case is economically disadvantageous.

In the present invention, various known active ingredients that are usually used, for example, retinol, retinoic acid known as an anti-aging agent, kojic acid known as a whitening agent, quercetin, glutathione, hydroquinone and derivatives thereof, condensation Type tannins, phenolic compounds such as caffeic acid and ellagic acid, and peripheral vasodilators such as vitamin E, vitamin E nicotinate, various vitamins such as nicotinic acid, nicotinic acid amide and benzyl nicotinate, ginger tincture and chili tincture As anti-inflammatory agents, various compounds such as corticosteroids, ε-aminocaproic acid, lysozyme chloride, glycyrrhizin, allantoin, and other animal and plant / microbe derived from placenta extract, licorice extract, purple root extract, lactic acid bacteria culture extract, etc. Various extracts and the like may impair the effects of the present invention With no range, it can be used by adding appropriate according to prevailing purpose.

Furthermore, in addition to these known active ingredients, the cosmetics of the present invention, in addition to basic ingredients such as fats and oils, as well as known moisturizers, preservatives, antioxidants, chelating agents, pH adjusters as necessary. Various additives such as fragrances and coloring agents can be used in combination as long as the effects of the present invention are not impaired.

EXAMPLES Next, the present invention will be described by way of examples, but these disclosures show preferred embodiments of the present invention and do not limit the present invention in any way.

<Production Example 1>
Add 40 kg of water to 10 kg of red mock, add 1 wt% sodium citrate, swell for 5 hours at 50 ° C., adjust to pH 4 with 10% lactic acid, 0.004 wt% cellulase (MP Biomedicals) (Trade name: CELLULASE) and stirred and extracted at 50 ° C. for 24 hours and filtered. Two times the weight of ethanol was added to the filtrate, and 50 kg of water was added to the resulting precipitate to dissolve it, followed by filtration.

<Production Example 2>
Add 40 kg of water to 10 kg of red mock, add 1 wt% sodium citrate, swell for 5 hours at 50 ° C., adjust to pH 5 with 10% lactic acid, 0.004 wt% cellulase (MP Biomedicals) (Trade name: CELLULASE), and stirred and extracted at 50 ° C. for 4 hours and filtered. Two times the weight of ethanol was added to the filtrate, and 50 kg of water was added to the resulting precipitate to dissolve it, followed by filtration.

<Production Example 3>
100 kg of water is added to 10 kg of akamoku, 5% by weight of sodium citrate is added, swollen at 40 ° C. for 5 hours, adjusted to pH 4 with 10% lactic acid, 0.1% by weight of cellulase (MP Biomedicals) (Trade name: CELLULASE) and stirred and extracted at 40 ° C. for 18 hours and filtered. Double the weight of ethanol was added to the filtrate, and 70 kg of water was added to the resulting precipitate for dissolution, followed by filtration.

<Production Example 4>
Add 20 kg of water to 10 kg of red mock, add 0.1 wt% sodium citrate, swell for 8 hours at 50 ° C., adjust to pH 4 with 10% lactic acid, and add 0.001 wt% cellulase (MP Biomedicals, product name: CELLULASE) was stirred and extracted at 50 ° C. for 16 hours and filtered. Two times the amount of ethanol was added to the filtrate, and 40 kg of water was added to the resulting precipitate to dissolve it, followed by filtration.

<Production Example 5>
Add 40 kg of water to 10 kg of red mock, add 1 wt% sodium citrate, swell for 5 hours at 50 ° C., adjust to pH 4 with 10% lactic acid, 0.01 wt% cellulase (MP Biomedicals) (Trade name: CELLULASE) and stirred and extracted at 50 ° C. for 24 hours and filtered. Two times the amount of ethanol was added to the filtrate, and 50 kg of water was added to the resulting precipitate for dissolution, followed by filtration.

<Test Example 1> Hyaluronidase Activity Inhibiting Action a) Test Method Samples (described later) obtained in Production Example 1 and Comparative Examples 1, 2, and 3 described above are represented by 0.1 M acetate buffer (pH 4.0). 50 μL of hyaluronidase (type IV-S from Bovin testis, SIGMA, 5,200 Units / mL) solution was added to 100 μL of the solution diluted to the concentrations shown in 1 and Table 2, and incubated at 37 ° C. for 20 minutes, followed by enzyme activation. 100 μL of an agent (compound 48/80, manufactured by SIGMA, 1 mg / mL) solution was added and incubated at 37 ° C. for 20 minutes.
Next, 250 μL of sodium hyaluronate (from rooster comb, manufactured by SIGMA, 0.4 mg / mL) solution was added and incubated at 37 ° C. for 20 minutes. After 20 minutes, 0.4M sodium hydroxide solution and boric acid solution were added in order, boiled on a water bath for 3 minutes, and after ice cooling, 3mL of 1% p-dimethylbenzaldehyde acetic acid solution (manufactured by Nacalai Tesque) was added. The mixture was incubated at 37 ° C. for 20 minutes, and the amount of N-acetylglucosamine released was determined from the absorbance at 585 nm.
As a positive control, sodium cromoglycate, which is known as a pharmaceutical having a strong hyaluronidase activity inhibitory action, was used. In the test, after preparing a 0.2% sodium cromoglycate solution stock solution with 0.1 M acetate buffer (pH 4.0), 6.3, 12.5, 25.0, 50.0 (v / v)% concentration And measured in the same manner as the sample solution.
The hyaluronidase activity inhibitory action was calculated by the inhibition rate obtained from the following formula.
As a control, 0.1 M acetate buffer (pH 4.0) was used instead of the sample, and purified water was used instead of the hyaluronidase solution as each blank.
Inhibition rate (%) = {(A−B) − (C−D)} / (A−B) × 100
A: Absorbance of the control solution at 585 nm
B: Absorbance of the control solution blank at 585 nm
C: Absorbance at 585 nm of the sample solution
D: Absorbance at 585 nm of the sample solution blank

<Comparative Example 1>
40 kg of water was added to 10 kg of akamoku, 1 wt% sodium citrate was added, and the mixture was stirred at 50 ° C. for 24 hours and filtered.

<Comparative example 2>
Add 40 kg of water to 10 kg of red mock, add 1 wt% sodium citrate, swell for 5 hours at 50 ° C, adjust to pH 4 with 10% lactic acid, and add 0.004% cellulase ((MP Biomedicals) (Product name: CELLULASE) and stirred and extracted at 50 ° C. for 24 hours, followed by filtration.

<Comparative Example 3>
40 kg of water was added to 10 g of red mock, 1 wt% sodium citrate was added, swollen at 50 ° C. for 5 hours, adjusted to pH 8 with 10% sodium hydroxide, 0.004% cellulase (MP Biomedicals (Trade name: CELLULASE), and extracted by stirring at 50 ° C. for 24 hours and filtered.

b) Test results Table 1 shows the measurement results of the hyaluronidase activity inhibition test. A concentration-dependent effect was observed in the akamoku extract of the present invention, and a hyaluronidase activity inhibitory action stronger than 0.2% sodium cromoglycate was confirmed. Further, as is apparent from the measurement results of the hyaluronidase activity inhibition test of Production Example 1 and Comparative Example 1 in Table 2, it was confirmed that the hyaluronidase activity inhibition rate dramatically increased by performing cellulase treatment, and Production Example 1 From the results of Comparative Example 2 and Comparative Example 2, it was confirmed that the hyaluronidase activity inhibitory action was maintained even if impurities were removed by adding alcohol.

<Test Example 2> Stability test over time of extract a) Test method After leaving Production Example 1 and Comparative Examples 1 and 2 at 45 ° C. for 1 month, the appearance and color of the solution were visually observed, and at OD 420 nm. The absorbance was examined.

b) Test results Table 3 shows the results of the stability test over time. No significant change was observed in any of the four test items even after one month in the akamoku extract, which was the product of the present invention prepared according to Production Example 1. On the other hand, Comparative Examples 1 and 2 have brown and seaweed-specific strong odors from the start of the test, and confirm that precipitation occurs after one month, and Comparative Example 3 is more than Comparative Example 1. The coloring was strong, and precipitation was confirmed after one month as in Comparative Examples 1 and 2. As is clear from these facts, it was confirmed that the pH and the addition of alcohol in the extraction process had a great influence on the stabilization of the raw material of the red peach mushroom extract.

<Test Example 3> Alcohol Stability Test in Simple System of Extract a) Test Method Prepare 20 wt% aqueous solutions of Production Example 1 and Comparative Examples 1 to 3, and add ethanol to this to a concentration of 50%. The sample was allowed to stand at room temperature for 10 days, and the occurrence of sediment and precipitation immediately after the addition and after 10 days was visually observed.

b) Test results Table 4 shows the results of the alcohol stability test for 50% ethanol. No occurrence of sediment or precipitation was observed in the extract of the present invention, which was the product of the present invention prepared in Production Example 1, either immediately after ethanol addition or after 10 days. On the other hand, in Comparative Examples 1 to 3, it was confirmed that a slight amount of suspended matter was already observed immediately after the addition, and in addition, in Comparative Examples 1 and 3, precipitation also occurred after 10 days. As is clear from the above, it is suggested that the stability of the akamoku extract in an alcohol preparation is greatly improved by performing an alcohol addition process in the extraction step.

<Test Example 3> Stability Test in Extract Formulation System a) Test Method Formulation Example 1 (in which the invention product of Production Example 1 was blended) and Production Example 1 in Formulation Example 1 were compared with Comparative Example 1, Comparative Example 2 or The skin lotion substituted for Comparative Example 3 (hereinafter referred to as Comparative Formulation Examples 1, 2 and 3) was allowed to stand at 45 ° C. for 1 month, and the appearance and coloring degree of the solution were examined by visual observation and sensory test.

b) Test results Table 5 shows the results of stability tests of Formulation Example 1 and Comparative Formulation Examples 1 to 3. In the lotion of Formulation Example 1, no change in appearance was observed even when left at 45 ° C. for 1 month. On the other hand, Comparative Formulation Examples 1 to 3 were colored slightly brown from the start of the test and had a slightly scent of seaweed peculiar to seaweed. Similarly, in Comparative Formulation Examples 1 to 3, an appearance close to brown was observed after 1 month, and a precipitate was also confirmed. The smell was enhanced and an unpleasant odor was produced. As is clear from these facts, it was confirmed that the treatment of pH and alcohol addition in the extraction process contributed to the stabilization of the preparation system.

Formulation Example 1 Lotion

Formulation Example 2 Essence

Formulation Example 3 Cream

Formulation Example 4 Cream Pack

これら処方例１乃至５は、いずれも本発明の目的を達成する効果を有していることが確認された。

Formulation Example 5 Latex

It was confirmed that all of these Formulation Examples 1 to 5 have the effect of achieving the object of the present invention.

Claims (1)

An anti-aging agent characterized in that the brown algae Sargassum akasaroku (Sargassum horneri) is extracted with water and an extract obtained by enzyme treatment using cellulase in the extraction treatment step is blended as an active ingredient .