JP6364281B2 - Preventing or improving skin aging - Google Patents

Description

  The present invention is useful for improving skin wrinkles, sagging and firming, tightening the skin, etc., mechanosensor activator, collagen gel contraction promoter, skin elasticity improving agent, skin tarmi prevention or improving agent, skin aging It relates to preventive or ameliorating agents.

  The skin is mainly divided into three layers: epidermis, dermis, and subcutaneous tissue. Among them, the dermis is extremely important for maintaining the structure of the skin, and it is made strong and flexible by fibers such as collagen and elastin. Forming. Proteins constituting connective tissue fibers such as collagen and elastin are synthesized / degraded by fibroblasts. Fibroblasts control the state of connective tissue by interacting with fibers such as collagen.

  When fibroblasts are embedded and cultured in a collagen gel under normal fibroblast culture conditions, the collagen gel contracts (Non-patent Document 1). Collagen gel contraction varies depending on the number of cells and the amount of serum in the medium, and becomes more noticeable as the number of fibroblasts in the gel increases and the amount of serum in the medium increases. However, such contraction does not occur at all in a collagen-only gel without fibroblasts or a gel in which floating cells such as lymphocytes are embedded.

The mechanosensor is a factor whose activity is changed by direct or indirect mechanical stimulation, and functions as a receptor for mechanical stimulation. For example, epidermal growth factor receptor (EGFR) is a factor that is phosphorylated and activated by mechanical stress (Non-Patent Documents 2 and 3). Angiotensin II type 1 receptor (ATR1) is a receptor for angiotensin II, but is known to be activated even in the absence of angiotensin II by mechanical stress (Non-patent Document 4). The P2Y receptor is activated by receiving ATP secreted extracellularly due to mechanical stress (Non-patent Document 5). The TRPV receptor is a subfamily of the TRP receptor family and is a Ca ²⁺ influx channel activated by mechanical stress. p130Cas is a substrate of the Src kinase family and is a factor that has been shown to be related to cell adhesion (focal adhesion) and essential for actin skeleton formation and cell migration (Non-patent Document 6). In recent years, it has been shown that it is a mechanosensor that senses physical force and enhances phosphorylation (Non-patent Document 7). Furthermore, Patent Document 1 describes that a collagen gel can be contracted by controlling the activity or expression of p130Cas.
In addition, regarding skin aging, it is known that fibroblasts derived from elderly people have reduced collagen gel contraction compared to cells derived from young people, and aging may reduce the ability of the gel to contract. It has become clear (Non-Patent Document 8). In other words, a decrease in the contractility of the collagen gel by fibroblasts is thought to indicate that the dermal connective tissue of the skin loses its contractile force, and further loses its strength and elasticity, resulting in wrinkles and tarmi. . Therefore, if the contraction ability of the collagen gel embedded with fibroblasts can be increased and the gel formation strength can be increased, the dermal connective tissue can be further contracted, tightened, and the strength can be increased. It is considered that the skin wrinkles and talmi caused by the above, and the reduced elasticity and firmness of the skin are improved, and the skin can be tightened. Under such circumstances, development of a collagen gel shrinkage accelerator has been desired.

  Conventionally, substances that promote the contraction of fibroblast-embedded collagen gel include serum, endothelin (Non-patent document 9), transforming growth factor β (Non-patent document 10, Non-patent document 11), platelet growth. Although growth promoting factors such as factors and compounds such as retinoic acid are known, there are problems with percutaneous absorption, stability, safety and price, and the effect is not sufficient.

On the other hand, Kogomebiyu (scientific name: Herniaria glabra) is a plant used for landscaping and the like because of its drought tolerance and rapid growth. It is also known to have a diuretic effect and is valuable as a medicinal herb. Recently, it has been reported that kogomebuyu has an effect of stimulating the expression of human antibacterial peptides, human β-defensin-2 (hBD2) and human β-defensin-3 (hBD3) (Patent Document 1). .
However, it has not been known so far that kogomebiyu has effects of activating p130Cas, promoting collagen gel contraction, improving skin elasticity, improving talmi and firmness, tightening skin, and improving wrinkles.

JP 2012-019698 A JP 2008-19264 A

E. Bell et al. J. Invest. Dermatol., 81, 2s (1983) Yano et al, J Invest Dermatol, 2004, 122, 783 Kippenberger et al, 2005, J Biol Chem, 280, 3060 Zou et al, 2004, Nat Cell Biol, 6, 499 Koizumi et al, Biochem J, 2004, 380, 329 Honda et al, 1998, Nat Genet, 19, 361, Huang et al, 2002, J Biol Chem, 277, 27265 Sawada et al, 2006, Cell, 127, 1015 M. Yamato, et al. Mech. Aging Dev. 67, 149 (1993) C. Guidry et al. J. Cell Biol., 115,873 (1991) R. Montesano et al. Pro. Natl, Acad. Sci. USA, 85, 4894 (1988) E. M. Grant et al. J. Cell Sci., 102, 315 (1992)

  The present invention provides a mechanosensor activator, collagen gel contraction promoter, skin elasticity improving agent, skin tarmi prevention or improvement agent, skin tightening agent, skin wrinkle prevention or improvement agent, skin aging prevention or improvement agent About doing.

  The inventors of the present invention have demonstrated that, from among highly safe plants, kogomebile or its extract activates p130Cas, which is a mechanosensor, and excellent collagen gel contraction promotion in a human skin fibroblast collagen-embedded culture system It has an action, and furthermore, it has been found to have an excellent elasticity improving action and wrinkle improving action on human skin, and this has improved skin elasticity, improved skin talmi and firmness, improved skin wrinkle, It was found useful for skin tightening, prevention or improvement of skin aging.

That is, the present invention relates to the following 1) to 7).
1) A mechanosensor activator comprising kogomebyu or an extract thereof as an active ingredient.
2) Collagen gel contraction promoter containing kogomebyu or its extract as an active ingredient.
3) Skin elasticity improving agent which uses kogomebyu or its extract as an active ingredient.
4) A preventive or ameliorating agent for skin talmi containing kogomebyu or its extract as an active ingredient.
5) A skin tightening agent comprising kogomebyu or an extract thereof as an active ingredient.
6) An agent for preventing or improving skin wrinkles comprising kogomebiyu or an extract thereof as an active ingredient.
7) A skin aging preventive or ameliorating agent comprising kogomebyu or an extract thereof as an active ingredient.

  Examples of the mechanosensor activator, collagen gel contraction promoter, skin elasticity improving agent, skin tarmi prevention or improvement agent, skin tightening agent, skin wrinkle prevention or improvement agent, skin aging prevention or improvement agent of the present invention include: By applying to the skin, the skin elasticity is improved, and the effect of suppressing or recovering tarmi, maintaining or recovering elasticity, skin tightening effect, skin wrinkle improving effect and the like are exhibited.

The figure which shows the p130Cas phosphorylation enhancement effect by a Kogomebyu extract.

  The “kogomebyu” of the present invention is also called a “Rupturewort Herb” and means Herniaria grabra belonging to the genus Kogomebyu.

  The kogomebyu of the present invention can be used in whole grass, leaves, stems, buds, flowers, buds, roots, rhizomes, seeds, or the like, or a combination thereof. Is preferred. Such kogomebyu can be used as a juice obtained by squeezing it as it is, a dried product obtained by drying the plant itself, or a pulverized product thereof, or an extract extracted from these. Is preferred.

As the extract, various solvent extracts obtained by a known extraction method, such as extraction of a plant at room temperature or under heating, or extraction using an extraction device such as a Soxhlet extractor, its diluted solution, A concentrated solution or a dry powder thereof may be mentioned.
Known extraction methods include, for example, immersion, decoction, leaching, reflux extraction, supercritical extraction, ultrasonic extraction, and microwave extraction.

As the extraction solvent used for obtaining the extract, either a polar solvent or a nonpolar solvent can be used. For example, water; alcohols such as methanol, ethanol, propanol and butanol; polyhydric alcohols such as propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; tetrahydrofuran and diethyl ether Linear and cyclic ethers such as polyethylene; polyethers such as polyethylene glycol; halogenated hydrocarbons such as dichloromethane, chloroform and carbon tetrachloride; hydrocarbons such as hexane, cyclohexane and petroleum ether; aroma such as benzene and toluene Group hydrocarbons; pyridines and the like, and these can be used alone or as a mixture. Of these, water, alcohol-based (alcohols and / or polyhydric alcohols) solvents, and water-alcohol mixed solvents are preferably used, and water-alcohol mixed solvents are preferable. Furthermore, as alcohols, methanol, ethanol, propanol, and butanol are more preferable, and ethanol is more preferable. Further, as the polyhydric alcohol, butylene glycol is more preferable.
More preferable water-alcohol mixed solvent is 20% (v / v) or more, preferably 50% (v / v) or more, and 95% (v / v) or less, 90% (v / v) or less. The ethanol aqueous solution of this is mentioned. For example, 20-95% (v / v) ethanol aqueous solution, Preferably 50-95% (v / v) ethanol aqueous solution is mentioned.

  The kogomebille extract of the present invention can be obtained, for example, by using 1 to 50 parts by mass of an extraction solvent with respect to 1 part by mass of a plant body and extracting at 4 to 100 ° C. for 0.5 to 30 days. . More specifically, when water is used as the extraction solvent, 5 to 30 parts by mass and 40 to 100 ° C. for 1 hour to 1 day are preferable with respect to 1 part by mass of the plant body. Moreover, when using a water-ethanol mixed solvent as an extraction solvent, 5-30 mass parts with respect to 1 mass part of plant bodies, 1 hour-20 days under room temperature-recirculation | reflux are preferable. Further, these operations may be repeated.

  The above extract can be used as it is, but it can also be used by diluting, concentrating or lyophilizing the extract and preparing it in a powder or paste form. Moreover, you may use a commercial item other than the said extraction processed material.

  The extract is preferably used after removing inactive impurities by a known technique such as liquid-liquid distribution, solid-liquid distribution, filtration membrane, activated carbon, adsorption resin, ion exchange resin, and the like. As the solvent used at this time, those exemplified for the extraction solvent may be used. Moreover, you may use these, after giving processes, such as a deodorizing and a decoloring, by a well-known method as needed. Moreover, when the extract is further purified, the known techniques and methods may be used.

As will be described later in Examples, the kogomebille extract of the present invention has an action of activating p130Cas, which is a mechanosensor, and an action of accelerating the contraction of a collagen gel embedded with human skin fibroblasts. Furthermore, it has an excellent elasticity improving effect and wrinkle improving effect on human skin.
As described above, it has been known that p130Cas can contract a collagen gel by controlling its activity or expression (Patent Document 1). Recently, a research group including the present inventors has shown that p130Cas has an activity of p130Cas. It decreased in aging skin, and it was shown that p130Cas is involved in aging of the skin (reference example described later, Table 5). Therefore, skin aging can be prevented or improved by activating p130Cas, which is a mechanosensor. Moreover, if the contractility of the collagen gel in which fibroblasts are embedded can be enhanced, it can be said that the dermal connective tissue can be contracted, tightened, and increased in strength. Therefore, the fact that kogomebille extract had p130Cas activating action means that kogomebyu or its extract is effective in preventing or improving skin aging, and the collagen gel embedded with human skin fibroblasts. The fact that it has a contraction-promoting action means that kogomebiyu or its extract can shrink the dermal connective tissue more, tighten, increase the strength, improve the skin elasticity reduced by aging, etc. It can be said to be effective for suppressing and recovering skin and wrinkles, maintaining or recovering elasticity, and tightening the skin.

Therefore, the kogomebille or extract thereof of the present invention is a mechanosensor activator, collagen gel contraction promoter, skin elasticity improving agent, skin tarmi prevention or improvement agent, skin tightening agent, skin wrinkle prevention or improvement agent. Alternatively, it can be an agent for preventing or improving skin aging (referred to as “mechanosensor activator and the like”), and can be used for producing a mechanosensor activator and the like. That is, the mechanosensor activator and the like of the present invention is applied to a human whose skin elasticity and firmness is reduced and is worried about tarmi and wrinkles to improve skin elasticity, prevention or improvement of skin tarmi, It can be used for tightening, prevention or improvement of skin wrinkles, or prevention or improvement of skin aging.
Here, the use for human may be therapeutic use or non-therapeutic use. “Non-therapeutic” means a concept that does not include medical practice, that is, a concept that does not include a method for surgery, treatment, or diagnosis of a human, more specifically, a doctor or a person who has received instructions from a doctor operates on a human. It is a concept that does not include a method of performing treatment or diagnosis.

  The mechanosensor activator is itself a cosmetic, pharmaceutical product for improving skin elasticity, preventing or improving skin talmi, tightening skin, preventing or improving skin wrinkles, or preventing or improving skin aging. It may be a quasi-drug, a pharmaceutical product, or may be a material or a preparation used by blending in the cosmetic, quasi-drug, pharmaceutical product or the like.

  In the present specification, the “mechanosensor” is a factor whose activity or expression is changed by mechanical stimulation. The mechanosensor may be a factor that senses mechanical stress (for example, contraction or extension) itself due to external force, or a factor that accepts a cellular signal that is secondarily activated by mechanical stress. Good. Preferred mechanosensors include, for example, epidermal growth factor receptor (EGFR), angiotensin II type 1 receptor (ATR1), P2Y receptor, TRPV receptor, p130Cas, their genes, or their mRNA. Of these, p130Cas is more preferable.

In the present specification, “collagen gel” means a gel-like structure containing collagen and fibroblasts in the dermis of a living body, or the extracellular matrix around it, and “collagen gel contraction” This means that the volume of the collagen gel is reduced.
“Improvement of skin elasticity” includes improvement of skin elasticity and suppression of skin elasticity reduction due to aging, etc., and “skin tarmi prevention or improvement” occurs along with skin elasticity reduction. Inhibition and recovery of skin tarmi, recovery and maintenance of elasticity, suppression of wrinkle reduction and wrinkle increase are included.
“Skin aging” includes either or both of physiological aging associated with aging and photoaging due to exposure to sunlight (ultraviolet rays), and mainly means that the skin is free of wrinkles and skin wrinkles and tarmi are generated.

Cosmetics, quasi-drugs or pharmaceuticals containing kogomebyu or its extract are preferably in the form of an external preparation for skin, specifically, ointments, emulsified cosmetics, creams, emulsions, lotions, gels, aerosols, etc. It can be used in various forms.
Such preparations are prepared according to general production methods, directly or pharmaceutically acceptable carriers such as various oils, surfactants, gelling agents, preservatives, antioxidants, solvents, alcohols, water, chelating agents. It can be obtained by mixing and dispersing together with a thickener, an ultraviolet absorber, an emulsion stabilizer, a pH adjuster, a pigment, a fragrance, and the like, and then processing into a desired form. In addition, for these cosmetics, quasi drugs or pharmaceuticals, etc., plant extracts, bactericides, moisturizers, anti-inflammatory agents, antibacterial agents, cooling agents, antiseborrheic agents, etc. Can be appropriately blended within a range not impeding the effects of the present invention.

  The content of kogomebyu or an extract thereof in the cosmetic, quasi-drug or pharmaceutical is generally 0.00001% by mass, preferably 0.001% by mass or less, and 10% by mass or less as a solid content concentration. Preferably, it is 1 mass% or less. Moreover, it is preferable to set it as 0.00001-10 mass%, and it is more preferable to set it as 0.001-1 mass%.

The dosage of the above cosmetics, quasi drugs or pharmaceuticals is not particularly limited as long as the effect is obtained, and may vary according to the condition, weight, sex, age or other factors of the subject. ) As per day, per day, as kogomebyu or its extract (in terms of dry matter), for example, it is preferably 0.5 mg or more, and 1000 mg or less, preferably 500 mg or less. Moreover, it is preferable to set it as 0.5-1000 mg, and also it is preferable to set it as 0.5-500 mg. The preparation can be ingested / administered according to any ingestion / administration plan, but is preferably divided into once to several times a day and continuously administered for several weeks to several months. Among these, it is more preferable to divide into 3 times a day and to administer continuously for 6 weeks or more.
In addition, the application target of the above cosmetics, pharmaceuticals or quasi drugs is not particularly limited as long as it is necessary, but it is improved skin elasticity, prevention or improvement of skin talmi and firmness, prevention of skin wrinkles. Or humans who desire improvement or skin tightening are preferred.

With respect to the above-described embodiment, the following aspects are disclosed in the present invention.
<1> A mechanosensor activator comprising kogomebiyu or an extract thereof as an active ingredient.
<2> The mechanosensor activator according to <1>, wherein the mechanosensor is p130Cas.
<3> Collagen gel contraction promoter comprising kogomebiyu or its extract as an active ingredient.
<4> A skin elasticity improving agent comprising kogomebiyu or an extract thereof as an active ingredient.
<5> An agent for preventing or improving skin talmi, comprising kogomebyu or an extract thereof as an active ingredient.
<6> A skin tightening agent comprising kogomebyu or an extract thereof as an active ingredient.
<7> A skin wrinkle prevention or ameliorating agent comprising kogomebyu or an extract thereof as an active ingredient.
<8> A preventive or ameliorating agent for skin aging comprising Kogomebyu or its extract as an active ingredient.
<9> Use of kogomebyu or an extract thereof for producing a mechanosensor activator.
<10> Use of kogomebyu or an extract thereof for producing a collagen gel contraction promoter.
<11> Use of kogomebyu or an extract thereof for producing a skin elasticity improving agent.
<12> Use of kogomebyu or an extract thereof for producing an agent for preventing or improving skin talmi.
<13> Use of kogomebyu or an extract thereof for producing a skin tightening agent.
<14> Use of kogomebyu or an extract thereof for producing an agent for preventing or improving skin wrinkles.
<15> Use of kogomebiyu or an extract thereof for producing an agent for preventing or improving skin aging.
<16> Kogomebyu or extract thereof for use in activating mechanosensors.
<17> Kogomebyu or extract thereof for use in promoting collagen gel contraction.
<18> Kogomebyu or an extract thereof for use in improving skin elasticity.
<19> Kogomebyu or an extract thereof for use in preventing or improving skin talmi.
<20> Kogomebyu or an extract thereof for use in tightening the skin.
<21> Kogomebyu or an extract thereof for use in preventing or improving skin wrinkles.
<22> Kogomebyu or an extract thereof for use in preventing or improving skin aging.
<23> A method for improving skin elasticity, comprising administering or ingesting kogomebyu or an extract thereof in an effective amount to a subject in need thereof.
<24> A method for preventing or improving skin tarmi, comprising administering or ingesting an effective amount of kogomebille or an extract thereof to a subject in need thereof.
<25> A skin tightening method of administering or ingesting an effective amount of kogomebille or an extract thereof to a subject in need thereof.
<26> A method for preventing or improving skin wrinkles, comprising administering or ingesting kogomebyu or an extract thereof in an effective amount to a subject in need thereof.
<27> A method for preventing or improving skin aging, comprising administering or ingesting an effective amount of kogomebiyu or an extract thereof to a subject in need thereof.
<28> In <16> to <27>, the use or method is a non-therapeutic use or non-therapeutic method.
<29> In <9> and <16>, the mechanosensor is p130Cas.
<30> In the above items <1> to <27>, the use site of the plant is preferably whole plant.
<31> In the above items <1> to <27>, the extraction solvent for obtaining the plant extract is water, an alcohol (alcohol and / or polyhydric alcohol) solvent, or a water-alcohol mixed solvent. . Here, methanol, ethanol, propanol, and butanol are more preferable as alcohols, ethanol is more preferable, and butylene glycol is more preferable as polyhydric alcohols.
<32> In the above items <1> to <27>, the extraction solvent for obtaining a plant extract is a water-alcohol mixed solvent, and more preferably 20% (v / v) to 95% (v / V) or less ethanol aqueous solution, 50% (v / v) or more and 95% (v / v) or less ethanol aqueous solution.
<33> In the above items <1> to <27>, an extraction solvent for obtaining a plant extract is a hydrocarbon, preferably hexane.

EXAMPLES Hereinafter, an Example is shown and this invention is demonstrated more concretely.
Example 1 Manufacture of Kogomebyu Water Extract 50 g of pulverized whole body of Kogomebyu (obtained from Monteagle Herbs) was immersed in 500 mL of water and extracted at 70 ° C. for 5 hours. After extraction, filtration was performed to obtain an extract. The extract was concentrated and dissolved in 10% ethanol so that the evaporation residue was 1 (w / v)% to prepare a water extract of kogomebille.

Example 2 Production of 50% ethanol extract of kogomebyu 50 g of pulverized whole plant of kogomebyu (obtained from Monteagle Herbs) was immersed in 500 mL of 50% ethanol aqueous solution and extracted at room temperature for 7 to 15 days. After extraction, filtration was performed to obtain an extract. The extract was concentrated and dissolved in 50% ethanol so that the evaporation residue was 1 (w / v)% to prepare a 50% ethanol extract of kogomebille.

Example 3 Production of 95% ethanol extract of kogomebyu 50 g of pulverized whole plant of kogomebyu (obtained from Monteagle Herbs) was immersed in 500 mL of 95% ethanol aqueous solution and extracted at room temperature for 7 to 15 days. After extraction, filtration was performed to obtain an extract. The extract was concentrated and dissolved in 95% ethanol so that the evaporation residue was 1 (w / v)% to prepare a 95% ethanol extract of kogomebille.

Example 4 Manufacture of hexane extract of Kogomebyu 10 g of pulverized whole of Komemebuyu (obtained from Monteagle Herbs) was immersed in 200 mL of hexane and extracted at room temperature for 7 to 15 days. After extraction, filtration was performed to obtain an extract. The extract was concentrated and dissolved in 99.5% ethanol so that the evaporation residue was 1 (w / v)% to prepare a hexane extract of kogomebille.

Test Example 1 Measurement of Collagen Gel Shrinkage Promoting Ability Collagen gel was prepared by referring to the literature “J. Cell Science, 102 , 315 (1992)” or “J. Invest. Dermatol, 93 , 792 (1989)”. . That is, HEPES (containing 0.01 M sodium hydroxide), DMEM (GIBCO DMEM, manufactured by Nitta Gelatin Co., Ltd., tape IA (3.0 mg / mL, pH = 3)) under ice-cooling. low glucose) 5-fold concentrated solution, FBS (5%, Fatal Bovine Serum), purified water, and after sufficiently neutralizing with stirring, the final concentration is 0.001 or 0.0003 mass% (dry solid mass%) Test substance (Kogomebyu extract prepared in Examples 1 to 4 or ethanol as a control) was added so that and finally a suspension of human dermal fibroblasts (derived from human foreskin) was added and stirred well, After removing bubbles, 0.6 mL of each hole was poured into a 24-well plate and immediately gelled at 37 ° C. The collagen concentration at this time was adjusted to 1.5 mg / mL. After 24 hours, liquid medium was added to each hole of the plate, the periphery of the gel was peeled off, and then further cultured at 37 ° C. for 48 hours. After incubation for 48 hours, the weight of the gel was measured with a balance.

  The results are also shown in Table 1. The value obtained by correcting the weight of the gel to which the kogomebyu extract was added with the weight of the control was expressed as the shrinkage acceleration rate. As apparent from Table 1, the addition of the kogomebyu extract reduced the weight, ie, volume, of the collagen gel and promoted the contraction of the collagen gel.

Test Example 2 Measurement of activating ability of p130Cas Human skin fibroblasts (derived from human foreskin, Cascade Biologics) at a cell density of 1 × 10 ⁵ cells / well on a 12-well plate, FBS (5%, Fatal Bovine Serum) Was seeded using a DME medium (GIBCO DMEM, high glucose). On the next day, the medium was replaced with FME-free DME medium, and one day later, the Kogomebille extract prepared in Example 3 was added to a final concentration of 0.001% (dry solid equivalent mass%); Replaced. In addition, the cell which added the equivalent amount of 95% ethanol which is a solvent of a Kogomebille extract was made into control. Further, the cells were collected on the next day, and then the protein was extracted according to a conventional method using Cell lysis buffer, and activated p130Cas and total p130Cas protein were detected by Western blotting using anti-phosphorylated p130Cas antibody and p130Cas antibody. The obtained target band was quantified by image analysis using Lane & Spot analyzer (ATTO).
The results are shown in FIG. As is clear from FIG. 1 and Table 2, the addition of Kogomebille extract increased phosphorylation of p130Cas and activated p130Cas.

Test example 3 Anti-aging effect test Half face comparison in the same subject using the serum formulation containing 5% of kogomebyu extract prepared in Example 3, with 9 male subjects and placebo as negative control A coating test was conducted. The subjects received a sample containing an appropriate amount (about 200 μL) of kogomebille extract on either the right or left side of the face and a prescription (placebo) added with a solvent instead of the extract three times a day (morning (Evening / Night) It was applied daily for 3 months. At the start of the test (at 0 week), and 6 and 12 weeks after the start of the test, visual evaluation of wrinkles on the corners of the eyes, photography, and skin elasticity measurement using a skin property measuring composite machine were performed.

(1) Evaluation of the ability to improve the elasticity of the human cheek The skin elasticity of the outer corners of the eyes and the cheeks was measured using a skin property measurement complex machine (MPA580, courage-khazaka). Table 3 shows the rate of change in pure elasticity (Uv / Ue) when the start time (0 week) is 1.
As is apparent from Table 3, the elasticity value was significantly higher in the application group of kogomebyu extract after 6 weeks and 12 weeks than before application, whereas there was no significant difference in the placebo application group. In addition, an excellent skin elasticity improving effect was observed with the kogomebyu extract.

(2) Evaluation of ability to improve human eye corner wrinkles Eye corners were photographed in accordance with the guidelines. That is, a strobe (Mono 200 Protect) and a digital camera-PC control system (Nikon D200, Nikon Capture, Nikon) were used. The subject sat on a chair and fixed his chin and face on a fixed base. Relative evaluation of the degree of wrinkles was performed using the photograph taken. Test according to the method of reference literature (Non-patent literature: Takeya M. et al, 2013. Comparisons between the relative evaluation method of wrinkles and the absolute scoring method of wrinkles using photos. Journal of Cosmetic Society 37, 268-73) The photographs taken at the start (week 0) and 6 weeks or at the end of the study (12 weeks) were compared, and the relative change in wrinkles after 6 weeks and 12 weeks after the start was determined as “2. Scoring was performed in five stages: “Clearly decreased”, “1. Slightly decreased”, “0. No change”, “−1. Slightly increased”, and “-2. Clearly increased”. The evaluation was conducted by researchers who have mastered the wrinkle evaluation equivalent to the trained expert of the guidelines established by the Cosmetic Society. In this evaluation, evaluation was performed under conditions that do not reveal to the evaluator whether the subject's left or right side is the sample side or the placebo side. The results are shown in Table 4.
As is apparent from Table 4, the wrinkle improvement application group had a greater degree of wrinkle improvement than the placebo application group, and an excellent wrinkle improvement effect was observed with the kogomebyu extract.

Reference Example 1 p130Cas activity decrease associated with skin aging Hairless mouse skin 4 weeks old and 24 weeks old or older was purchased from Takara Bio Inc. Skin fragments were crushed in a cell lysis buffer (Cell Signaling) using a glass homogenizer, and proteins were collected. Western blotting analysis was performed according to a conventional method. Phosphorylated activated p130Cas and total130Cas were detected using an anti-phosphorylated p130Cas antibody (Cell Signaling) and a p130Cas antibody (BD Bioscience). Table 5 shows the results obtained by quantifying the obtained target band by image analysis using Lane & Spot analyzer (ATTO) and correcting the quantified value of activated p130Cas with the quantified value of totalp130Cas.
From the results in Table 5, since the phosphorylation level of p130Cas decreases in aging skin, it is considered that the activity of p130Cas decreases with skin aging.

Claims (6)

  A mechanosensor activator comprising kogomebyu or an extract thereof as an active ingredient.   The mechanosensor activator according to claim 1, wherein the mechanosensor is p130Cas.   Collagen gel contraction promoter containing kogomebyu or its extract as an active ingredient.   A skin elasticity improving agent comprising kogomebyu or an extract thereof as an active ingredient.   An agent for preventing or improving skin talmi comprising kogomebyu or its extract as an active ingredient.   A skin tightening agent comprising kogomebyu or its extract as an active ingredient.