JP5164198B2 - Profilaggrin production promoter, filaggrin production promoter and pore conspicuousness inhibitor - Google Patents

Description

  The present invention relates to a profilagrine production promoter, a filaggrin production promoter and a skin pore conspicuousness inhibitor.

  An amino acid which is a main component of natural moisturizing factors (NMF) is produced by degrading filaggrin derived from keratohyalin granules in the stratum corneum. This filaggrin is expressed as profilagrin in epidermal keratinocytes present in the granule layer immediately below the stratum corneum. It is then phosphorylated immediately, accumulated in keratohyalin granules, decomposed to filaggrin through dephosphorylation and hydrolysis, transferred to the stratum corneum, increased the efficiency of keratin filament aggregation, and is involved in the internal construction of keratinocytes It is known (see Non-Patent Document 1).

  In recent years, it has been known that this filaggrin is very important and indispensable for moisture retention of the skin, and that the synthetic power of filaggrin is reduced by conditions such as drying, and the amount of amino acids in the stratum corneum is reduced ( Non-patent document 2).

  Therefore, in epidermal keratinocytes, it is expected that the water environment of the stratum corneum can be essentially improved by promoting the production of filaggrin through the promotion of profilagrin mRNA expression and thereby increasing the amount of amino acids in the stratum corneum. .

  Examples of such profilagrin production promoter and filaggrin production promoter include licorice extract (see Patent Document 1), liquiritin known as flavanone glycoside contained in natural plants (see Patent Document 2), pro Known as filaggrin and filaggrin protein production promoters are plant extracts or yeast extracts belonging to the genus Citrus (see Patent Document 3).

  In addition, the conspicuous skin pores is one of the skin concerns for women. The conspicuous pores in the skin are caused not only by the size of the pores themselves but also by square plugs, dents around the pores, uneven color, and the like.

  Therefore, in order to suppress the conspicuous pores, square plug removal cosmetics (so-called pore packs) and pore shrinking agents are used (see Patent Document 4), but generally powder cosmetics such as foundation are applied. In this way, apparently pores are made inconspicuous.

However, consumer demands for profilagrin production promoters, filaggrin production promoters, and pore-spot suppressors that have profilagrin production-promoting action, filaggrin production-promoting action, and pore-squeezing inhibitory action, and are inexpensive and highly safe are The current situation is that there is a strong demand for the development and provision of strong new profilaggulin production promoters, filaggrin production promoters and pore-spot suppressors.
JP 2002-363054 A JP 2003-146886 A JP 2001-261568 A JP 2002-187817 A "Fragrance Journal Extra Edition", 2000, Vol.17, p.14-19 "Arch. Dermatol. Res.", 1996, Vol.288, p.442-446

  The first object of the present invention is to find a profilagrin production promoting agent from among highly safe natural products, and to provide a profilagrin production promoter containing the same as an active ingredient.

  The second object of the present invention is to find a filaggrin production promoting action from highly safe natural products, and to provide a filaggrin production promoter comprising this as an active ingredient.

  Thirdly, an object of the present invention is to find a pore safety inhibitor from natural products having high safety, and to provide an agent for suppressing pore visibility, which uses it as an active ingredient.

  In order to solve the above-mentioned problems, the present inventors have conducted intensive studies. As a result, Agez extract has an excellent profilagrin mRNA expression promoting effect, and the production of filaggrin through profilaggrin mRNA expression promoting effect in epidermal cells. It has been found that it can prevent and improve dry skin, wrinkles, rough skin, and atopic dermatitis by promoting it, and also has the effect of making the dents around the pores shallower and smaller, and reducing the volume of the pores themselves. I found out that there was.

  The present invention is based on such findings by the present inventors, and the profilagrin production promoter, filaggrin production promoter and pore pore conspicuousness inhibitor of the present invention contain an extract from Agetsu as an active ingredient. Features.

  Here, in the present invention, “suppression of pore conspicuity” means to make the pores inconspicuous by reducing the volume of the pores in the skin, to reduce the size of the pores themselves, and to dent the skin around the pores. Including improvements.

  ADVANTAGE OF THE INVENTION According to this invention, the extract from the natural product Ages can be provided, and the profilagrin production promoter, filaggrin production promoter, and pore conspicuousness inhibitor which are highly safe can be provided.

  The profilaggrin production promoter, filaggrin production promoter and pore-remarking inhibitor of the present invention contain Ages extract as an active ingredient.

  In the present invention, the “extract from Ages” refers to an extract obtained by using Ages as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or a crude product thereof. Either a purified product or a purified product is included.

  The extraction raw material used in the present invention is Ages. Ages refers to dried mature fruits of the rose (Rosa multiflora), a Rosaceae plant. Neurose (Rosa multiflora) is a deciduous shrub distributed from southwestern Hokkaido to Kyushu and the Korean peninsula. Ages, the dried product of mature roses, can be easily obtained from these areas.

  The details of the substance having a profilagrine production promoting action, filaggrin production promoting action or pore conspicuousness suppressing action contained in the extract from Ages are unknown, but depending on the extraction method generally used for plant extraction, From these, an extract having these actions can be obtained.

  For example, after drying the above plant, it can be obtained by pulverizing as it is or using a crusher and subjecting to extraction by solvent extraction to obtain an extract having a profilagline production promoting action, a filaggrin production promoting action or a pore conspicuousness inhibiting action. it can. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing pretreatment such as degreasing, the extraction treatment with Ages polar solvent can be performed efficiently.

  As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water and hydrophilic organic solvents. These may be used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. Is preferred.

  Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in the present invention includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using a liquid mixture of water and a lower aliphatic alcohol, it is preferable to mix 1 to 90 parts by weight of a lower aliphatic alcohol with respect to 10 parts by weight of water. When using a mixed solution, it is preferable to mix 1 to 40 parts by mass of a lower aliphatic ketone with 10 parts by mass of water, and when using a mixed solution of water and a polyhydric alcohol, water 10 It is preferable to mix 10 to 90 parts by mass of polyhydric alcohol with respect to parts by mass.

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent 5 to 15 times (mass ratio) of the extraction raw material, the soluble components are extracted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. The obtained extract is diluted, concentrated, dried, purified, etc. according to a conventional method in order to obtain a diluted or concentrated solution of the extract, a dried product of the extract, or a crude purified product or a purified product thereof. Processing may be performed.

  Purification can be performed by, for example, activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, or the like. The obtained extract can be used as an active ingredient of a profilagrin production promoter, a filaggrin production promoter or a pore conspicuousness inhibitor as it is, but a concentrated solution or a dried product is easier to use.

  Since the extract from Agetsu has a unique odor, it can be purified for the purpose of decoloring, deodorizing, etc. as long as it does not cause a decrease in its physiological activity, but it is incorporated into skin cosmetics etc. In this case, since it is not used in large quantities, there is no practical problem even if it is not purified.

  Since the extract from Agez obtained as described above has a profilagrine production promoting action, a filaggrin production promoting action or a pore conspicuousness inhibiting action, the profilagline production promoting agent, filaggrin is utilized by utilizing each action. It can be used as an active ingredient of a production promoter or a pore-reducing agent.

  The profilaggrin production promoter, filaggrin production promoter or pore-remarking inhibitor of the present invention may consist only of an extract from Ages, or may be a product of the above extract.

  The extract from Ages is in a dosage form such as powder, granule, tablet, liquid, etc. according to a conventional method using a pharmaceutically acceptable carrier such as dextrin and cyclodextrin and any other auxiliary agent. It can be formulated. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring agent and the like can be used. The extract from Agez can be used by blending with other compositions (for example, skin cosmetics, etc.), and can also be used as an ointment, a liquid for external use, a patch, and the like.

  In addition, the profilaggulin production promoter, filaggrin production promoter or pore conspicuousness inhibitor of the present invention, if necessary, other natural extracts having a profilaggrin production promoting action, filaggrin production promoting action or pore conspicuousness inhibiting action It can mix | blend and can be used as an active ingredient.

  The profilaggrin production promoter of the present invention promotes the production of profilaggrin in the cell through the profilaggrin production promoting action of the extract from Agetsu, and increases the amount of filaggrin obtained by hydrolysis of profilagrin, The skin moisturizing ability can be improved, whereby the elasticity of the skin can be maintained, and skin aging, dry skin, wrinkles, rough skin, etc. can be prevented and improved. However, the profilaggrin production promoter of the present invention can be used for all purposes that are meaningful for exerting a profilagrin production promoting action in addition to these uses.

  The filaggrin production promoter of the present invention can promote the production of filaggrin in cells through the filaggrin production promoting action of the extract from Ages, and can improve the moisture retention ability of the skin, thereby improving the elasticity of the skin. It can maintain sexuality, and can prevent and improve skin aging, dry skin, wrinkles, rough skin, and the like. However, the filaggrin production promoter of the present invention can be used for all purposes that are meaningful for exerting the filaggrin production promoting action in addition to these uses.

  The pore conspicuousness inhibitor of the present invention can make pores inconspicuous through the pore conspicuousness suppressing action of the extract from Ages. However, the pore conspicuousness inhibitor of the present invention can be used for all purposes other than these uses, which are meaningful for exhibiting the pore conspicuousness inhibiting action.

  In addition, the profilaggulin production promoter, filaggrin production promoter, and pore conspicuousness suppressant of the present invention are preferably applied to humans, but as long as the respective effects are exhibited, animals other than humans It can also be applied to.

  Hereinafter, although a test example is shown and this invention is demonstrated concretely, this invention is not limited to the following test example at all. In this example, a freeze-dried product of Agetsu Extract-R (manufactured by Maruzen Pharmaceutical Co., Ltd.) was used as a sample.

[Example 1] Profilaggrin and filaggrin production promoting action test Normal human newborn skin epidermal keratinocytes (NHEK) in an 80 cm ² flask in normal human epidermal keratinocyte medium (KGM) at 37 ° C, 5% CO _2- The cells were cultured under the condition of 95% air and the cells were collected by a conventional method. The obtained cells were adjusted to a cell density of 1.5 × 10 ⁵ cells / mL in the same medium, seeded on a 6-well collagen-coated plate in a volume of 2 mL, and incubated at 37 ° C., 5% CO ₂ -95% air. For 4 days. After the culture, the medium was replaced with ₂ mL of KGM containing a sample dissolved in 0.25% DMSO (see Table 1 below for the sample concentration), and cultured for 5 days under conditions of 37 ° C. and 5% CO ₂ -95% air. . After completion of the culture, total protein was prepared by a conventional method.

<Western blotting>
The sample adjusted to 10 μg / row was developed by SDS-PAGE and transferred to a PVDF membrane. After blocking with PBS (−) containing 5% skim milk, anti-human filaggrin monoclonal antibody (Harbor Bio-Products), biotin-labeled anti-mouse Ig (Whole Ab, manufactured by Amersham Biosciences) and streptavidin-peroxidase complex ( CALBIOCHEM) was diluted 1000-fold with PBS (-) containing 0.1% Tween20 and 0.3% skim milk, and allowed to react sequentially, and luminescence from ECL Western blotting detection reagents and analysis system (Amersham Biosciences) Was used to detect profilaggrin and filaggrin. The detected band was quantitatively measured with KODAK 1D Image Analysis Software EDAS290 Version3.5.

  As a result, the profilaggrin production promoting effect of the sample was evaluated using the value obtained by adding the net intensity (band intensity) of profilagrin and filaggrin in 10 μg of the protein prepared from each of the cells cultured with and without the sample added. The profilaggrin production promotion rate (%) was calculated based on the following formula.

Profilaggrin production promotion rate (%) = A / B × 100
In the above formula, A represents “Net intensity at the time of sample addition (total value of profilagrin and filaggrin)” and B represents “Net intensity at the time of no sample addition (control)”.
The test results are shown in Table 1.

  As shown in Table 1, it was confirmed that the extract from Agetsu has an excellent profilagline production promoting action. Moreover, it was confirmed that the profilaggrin production promotion effect which the extract from Ages has depends on the concentration of the extract. In addition, since filaggrin is produced by hydrolysis of profilagrin in vivo, an extract from Ages having profilagrin production promoting action promotes the production of profilagrin, and profilagrin in the cell By increasing the amount, the amount of filaggrin can also be increased, and as a result, it is considered to have a filaggrin production promoting action.

[Example 2] Pore conspicuousness inhibitory test <Test site>
Both testers' left and right cheeks.
<Sample application method>
The amount of application at one time is about 0.3 mL, and in the morning and after bathing twice a day, control lotion (Ages extract-R no additive) and 1% Agez extract-R combination makeup are applied to the designated test site. Each with water was applied for 30 days.

<Measurement method>
The makeup was removed using a sheet-like makeup remover, washed with a tissue soaked with purified water, and placed in a constant temperature and humidity chamber (temperature 24 ° C., humidity 50%). After 30 minutes from entering the room, a replica of the test site was collected with a replica creation kit ASB-01 (manufactured by Asahi Biomed). The replica pore volume ratio (pore volume / surface area of the test site, μm ³ / mm ² ) before the start of the test and at the end of the test in the 1% Ageits extract-R-containing skin lotion application part, the 3D replica analysis system for reflection (Asahi Biomed Co., Ltd.) was used to evaluate the pore-reducing effect.
The test results are shown in Table 2. In Table 2, the pore volume ratio is shown as an average value of all the subjects (eight persons).

[Table 2]

  As shown in Table 2, it was confirmed that the volume of the pores can be reduced by applying the age extract, thereby making the pores inconspicuous.

  The profilaggrin production promoter and filaggrin production promoter of the present invention can greatly contribute to the improvement of the moisturizing function of the skin by promoting the production of profilagrin and filaggrin, and the pore conspicuousness suppressor of the present invention Can greatly contribute to the improvement of skin-related problems such as conspicuous pores.

Claims (2)

  A profilagulin production promoter comprising an age extract as an active ingredient.   A filaggrin production promoter characterized by containing Agetsu extract as an active ingredient.