JP7828033B2 - composition - Google Patents

Description

The present invention relates to a composition containing moutan pith and taurine.

Mitochondria are intracellular organelles whose primary function is to produce the energy ATP. Therefore, maintaining mitochondrial function contributes to maintaining cellular and ultimately bodily functions. It has become clear that mitochondrial function is regulated by mitochondrial fusion and fission, also known as mitochondrial dynamics, and interactions with other intracellular organelles. MITOL, a ubiquitin ligase localized in the outer mitochondrial membrane, has been reported to be involved in mitochondrial dynamics and the regulation of mitochondrial-endoplasmic reticulum adhesion (Non-Patent Document 1). Mice with cardiac-specific MITOL deficiency exhibited reduced cardiac function, as well as increased lipofuscin deposition and SA-β-gal expression, both of which are signs of cardiac aging (Non-Patent Document 2). Furthermore, mice lacking MITOL specifically in the epidermis exhibited hair follicle aging symptoms such as graying and hair loss, as well as skin aging-like findings such as epidermal thickening (Non-Patent Document 3). Furthermore, Alzheimer's disease model mice lacking MITOL specifically in the nerves exhibited worsening Alzheimer's disease pathology (Non-Patent Document 2). These findings suggest that MITOL loss is involved in the promotion of a wide range of aging symptoms, including cardiac aging, hair follicle aging, skin aging, and brain aging. Furthermore, it has been reported that MITOL knockdown in HELA cells elevated the expression of the senescence marker SA-β-gal, which was rescued by MITOL knock-in (Non-Patent Document 4). These findings suggest that MITOL loss accelerates aging, and that restoring reduced MITOL can reverse aging. Based on these findings, approaches that increase or activate MITOL production are considered useful for anti-aging.

To date, plant extracts such as those from Coptis japonica and Moutanga pisi (Patent Document 1) and berberine (Patent Document 2) are known to increase MITOL production, but there is a demand for materials with even stronger effects.

Increasing the concentration of each ingredient can be expected to produce a proportionally stronger effect, but using plant extracts and other ingredients at too high a concentration can lead to formulation issues such as discoloration, reduced stability and usability, and there is also the possibility of safety issues such as skin irritation, so simply increasing the amount used may not be desirable.

Nagashima Shun et al. (2014) Biochemistry, 86(1):63-67 Nagashima Shun et al. (2017) Japanese Pharmacological Journal, 149(6):254-259 Shigeru Yanagi (2016) Cosmetology Research Report, 24:149-152 Park et al. J Cell Sci. 2010;123:619-26.

Japanese Patent Application Laid-Open No. 2019-52145 Japanese Patent Application Laid-Open No. 2019-178070

The object of the present invention is to provide a composition that has excellent effects of inhibiting MITOL reduction or promoting MITOL production.

As a result of extensive research, the inventors discovered that combining Moutan Pea with taurine enhances the MITOL production-promoting effect of Moutan Pea, leading to the completion of the present invention.

That is, the present invention provides:
(1) A composition comprising Peony root and taurine;
(2) The composition according to (1), which is for external use on the skin.
(3) The composition according to (1) or (2), which is used to inhibit MITOL reduction or promote MITOL production.

By inhibiting MITOL reduction or promoting MITOL production, the composition of the present invention can prevent or improve aging symptoms, such as hair follicle aging symptoms such as gray hair and hair loss, and skin aging symptoms such as decreased skin barrier function and age spots and wrinkles. It can also be used to prevent or improve cardiac aging symptoms such as decreased cardiac function, and brain aging symptoms such as Alzheimer's disease. Furthermore, it can be used as a reagent to promote MITOL production, and is preferably used as a positive control drug when conducting material screening, etc.

FIG. 1 is a graph showing the results of a test evaluating the MITOL production promoting effect of Moutan Pea extract. FIG. 2 is a graph showing the results of a test example in which the MITOL production-promoting effect was evaluated when either Peony extract or taurine (20 mM) was used alone or in combination.

The term "peony" as used herein refers to the root bark of the tree peony (scientific names: Paeonia suffruticosa, Paeonia moutan) of the Paeoniaceae family.
Moutan Pea is used in the form of a herbal powder or herbal extract. The herbal powder used in the present invention may be, for example, a dried product obtained by further pulverizing a dried, chopped product into a fine powder. The Moutan Pea extract used in the present invention can be extracted with solvents such as water, lower aliphatic alcohols (e.g., methanol, ethanol, isopropyl alcohol), polyhydric alcohols (e.g., propylene glycol, 1,3-butylene glycol, glycerin, dipropylene glycol), and lower aliphatic ketones (e.g., acetone). However, extraction with a mixture of ethanol and water or a mixture of polyhydric alcohols and water is preferred, with a mixture of ethanol and water being most preferred. When extracting with a solvent consisting of water and ethanol, the ethanol content in the solvent is preferably 50-95% by volume. The extract form is not particularly limited; it can be prepared into a dried extract powder, extract powder, soft extract, or liquid extract by heat treatment, freeze-drying, or vacuum drying. It is also possible to use extracts purchased from various reagent and raw material manufacturers. Commercially available products of this type of Peony extract include "Peony Extract" and "Peony Extract BG" from Maruzen Pharmaceuticals, and "Falcorex (registered trademark) Peony B" and "Falcorex Peony E" from Ichimaru Falcos. The label name for quasi-drugs and cosmetics is Peony Extract or Peony Extract.

The amount of moutan pith in the composition of the present invention is not particularly limited, but is preferably 0.00000003 to 0.3 mass% in terms of solids, more preferably 0.000003 to 0.2 mass%, and even more preferably 0.00003 to 0.03 mass%.

Taurine (also known as aminoethylsulfonic acid or 2-aminoethanesulfonic acid) in this invention is a compound specified by Cas No. 107-35-7. Taurine can be produced by standard chemical synthesis or by extraction from seafood, particularly mollusks (octopus and squid). It can also be purchased from various reagent and raw material manufacturers.

The amount of taurine in the composition of the present invention is not particularly limited, but is preferably 0.0001 to 10% by mass, and more preferably 0.001 to 5% by mass.

In the composition of the present invention, the mass ratio of the content of Moutan Pea to taurine (Moutan Pea (solids equivalent): taurine) is not particularly limited, but from the perspective of inhibiting MITOL reduction or promoting MITOL production, a range of 1:0.3 to 1:30000 is preferred, 1:3 to 1:300 is more preferred, and a range of 1:50 to 1:300 is even more preferred.

Compositions of the present invention include pharmaceuticals, quasi-drugs, cosmetics, foods and beverages, reagents, etc. The dosage form is not particularly limited, but includes topical and oral administration, preferably topical application to the skin, including the scalp, and/or hair. When the present invention is applied topically to the skin, examples of dosage forms include lotions, liquids, creams, ointments, gels, sprays, shampoos, conditioners, and soaps. When the present invention is applied orally, examples of dosage forms include tablets, powders, granules, liquids, capsules, dry syrups, jellies, liquid foods, semi-solid foods, and solid foods.

These can be manufactured by known methods. During manufacturing, various additives that can be contained in cosmetics, quasi-drugs, pharmaceuticals, foods and beverages, or reagents can be blended within the scope that does not impair the effects of the present invention.

The composition of the present invention containing Moutan Peony and taurine has the effect of inhibiting the decrease in MITOL or promoting the production of MITOL, and based on this effect, it can prevent or improve aging symptoms such as hair follicle aging symptoms such as gray hair and hair loss, and skin aging symptoms such as a decrease in skin barrier function and age spots and wrinkles. It can also be used to prevent or improve cardiac aging symptoms such as decreased cardiac function, and brain aging symptoms such as Alzheimer's disease.

Mice lacking MITOL specifically in the skin epidermis have been observed to exhibit symptoms of hair follicle aging, such as increased gray hair, hair loss, and increased hair follicle cell death (Non-Patent Document 3). Hair pigment is produced by melanocytes present in the hair bulb of the hair follicle, which is the hair tissue. Hair follicle keratinocytes, such as hair matrix cells, are supplied with this pigment and form the resulting pigmented hair. When hair is replaced, the area below the bulge region of the hair follicle, such as the old hair bulb, disappears, and with it, the melanocytes in the hair bulb. When a new hair cycle begins and new hair is produced, melanocytes are supplied from melanocyte stem cells present in the bulge region of the hair follicle, resulting in the regeneration of pigmented hair. Aging and various stresses can cause a decrease or depletion of melanocyte stem cells, or problems with the melanocyte supply process, resulting in an inadequate supply of melanocytes, resulting in the development of pigment-depleted gray hair.

Hair follicle epithelial stem cells (hereafter referred to as "hair follicle stem cells") reside adjacent to melanocyte stem cells in the bulge region. When body hair grows out, hair follicle stem cells supply hair matrix cells, which are the epithelial cells of the hair follicle, and these cells form new hair follicle tissue. Furthermore, it has been revealed that hair follicle stem cells not only have the function of reconstructing the epithelial cells of the hair follicle, but also have the function of forming a microenvironment (hereafter referred to as "niche function") to maintain the adjacent melanocyte stem cells (Tanimura S et al (2011) Cell Stem Cell., 6(2):130-40.). Hair follicle stem cells maintain melanocyte stem cells by controlling the maintenance and activation of the undifferentiated state of melanocyte stem cells through TGFβ signaling and WNT signaling. Based on this, it is thought that the decline in melanocyte stem cells occurs when damage to hair follicle stem cells accumulates due to aging and various stresses, resulting in a loss of hair follicle stem cell niche function (Aoki H et al (2013) J Invest Dermatol., 133(9):2143-51.).

It has been elucidated that type XVII collagen plays an important role in maintaining hair follicle stem cells. Type XVII collagen is a bell-shaped collagen that makes up the hemidesmosomes (hemidesmosomes) that anchor epithelial cells to the basement membrane, and COL17A1 is a factor that makes up its alpha chain. Hair follicle stem cells are also anchored to the basement membrane by this hemidesmosome structure, and a deficiency in type XVII collagen leads to hair loss due to impaired hair follicle stem cell maintenance. This results in the loss of melanocyte stem cells and the graying of body hair. Furthermore, it has recently been elucidated that type XVII collagen is degraded with age, leading to the loss of hair follicle stem cells and hair loss (Matsumura H et al. (2016) Science., 351(6273):aad4395.). Based on these findings, promoting the production of COL17A1, a factor that makes up the alpha chain of type XVII collagen, and increasing the expression of type XVII collagen can prevent and improve hair loss and graying.

Mice in which SCF is forcibly expressed in epithelial cells, including hair follicle stem cells, have been found to be resistant to the stress that causes hair to turn gray. This suggests that increasing SCF expression in epithelial cells leads to the maintenance of melanocyte stem cells, resulting in the prevention and improvement of hair turning gray (Aoki H et al. (2011) J Invest Dermatol. 131(9):1906-15.).

WNT7A, a type of WNT ligand, is strongly expressed in hair follicles during the hair cycle. Body hair that regenerates after wound healing is usually white and lacks pigment. However, if the wound occurs during the anagen phase, when epithelial cells strongly express WNT7A, colored hair regenerates. This is due to WNT7A activating melanocyte stem cells. Therefore, increasing WNT7A expression can prevent and improve graying (Yuriguchi M et al (2016) J Dermatol Sci., 84(1):80-87.).

In addition, hair loss and graying can occur due to the decline in function or cell death of hair follicle keratinocytes, hair follicle stem cells, melanocyte stem cells, etc. Cell proliferation markers such as the cell cycle-related nuclear protein KI67 and the cell cycle-regulating protein PCNA serve as indicators of the activation of these cells, so increasing the expression of KI67, PCNA, etc. can prevent and improve hair loss and graying.

It has been confirmed that MITOL is involved in the proliferation of hair follicle stem cells and hair follicle keratinocytes through the expression of MKI67 and PCNA, in the expression of type XVII collagen through the expression of COL17A1, and in the maintenance of melanocyte stem cells through the expression of WNT7A, TGFβ2, and SCF (Patent Document 1). Therefore, ingredients that promote MITOL production or inhibit the decrease of MITOL have the effect of inhibiting the decrease, promoting the proliferation, and activating hair follicle stem cells, melanocyte stem cells, and hair follicle keratinocytes, and based on these effects, can provide effects for preventing and improving hair loss and graying (Patent Document 1).

Skin is broadly divided into three layers: the epidermis, dermis, and subcutaneous tissue. The main constituent cells of the epidermis are keratinocytes, which undergo four stages of morphological change during differentiation. Basal cells, the lowest layer, contact the basement membrane, synthesize DNA, and divide and proliferate. These divided cells migrate upward and become spinous cells. Further differentiation leads to granular cells, which ultimately differentiate into corneocytes, forming the stratum corneum (stratum corneum), the top layer. Keratinocytes at these stages of differentiation are layered within the epidermis. The stratum corneum formed in this way functions as a barrier to prevent various external stimuli from penetrating the body (skin barrier function). The keratinocytes that make up the stratum corneum are adhered to each other by desmosomes. However, in healthy skin, this adhesion mechanism gradually weakens, leading to spontaneous detachment of the stratum corneum in response to physical stimuli (stratum corneum desquamation). In this way, the epidermis undergoes a rhythmic cycle of proliferation, differentiation, and exfoliation (epidermal turnover). In addition to keratinocytes, the epidermis also contains pigment cells, which produce melanin. Normally, melanin is transferred to keratinocytes and excreted as the stratum corneum peels during the process of epidermal turnover. However, if there is an abnormality in epidermal turnover, such as a decrease in differentiation and proliferation from basal cells, the melanin is not excreted and accumulates in the epidermis, resulting in the formation of age spots.

Below the basal cells of the epidermis is the dermis, which consists of three layers: the papillary layer, the subpapillary layer, and the reticular layer. Fibroblasts are present in the dermis and synthesize extracellular matrix materials such as collagen fibers (collagenous fibers), elastin fibers (elastic fibers), and hyaluronic acid. Collagen fibers form thick fiber bundles in the reticular layer, making up the support tissue of the skin. Elastin fibers exist mixed with collagen fibers, giving the tissue elasticity. Hyaluronic acid has excellent moisture retention properties and provides flexibility to the skin.

As we age, the proliferation function of basal cells in the epidermis declines, resulting in thinning of the epidermis and a longer turnover time. Fibroblast proliferation also declines in the dermis, and the ability to synthesize extracellular matrix components also declines. As a result, the overall amount of collagen fibers decreases, becoming thinner and the gaps between fibers becoming larger. Furthermore, elastin fibers denature and lose their elasticity, the amount of hyaluronic acid also decreases, and moisture retention decreases. This results in atrophy of the skin, a loss of firmness, and other problems, leading to wrinkles and sagging. Furthermore, abnormalities in epidermal turnover can lead to the formation of age spots.

MITOL has been confirmed to be involved in the proliferation of epidermal keratinocytes, dermal fibroblasts, hair follicle keratinocytes, etc. through the expression of MKI67, PCNA, CDC25C, etc. in these cells (Patent Document 1). Therefore, ingredients that promote MITOL production or inhibit the reduction of MITOL have the effect of inhibiting the reduction, promoting the proliferation, and activating epidermal keratinocytes, dermal fibroblasts, and hair follicle keratinocytes, thereby preventing and improving skin aging symptoms such as a decrease in skin barrier function and age spots and wrinkles (Patent Document 1).

In addition, external factors such as ultraviolet rays are known to cause symptoms similar to chronological aging, also known as photoaging. UV rays and other factors increase reactive oxygen species (ROS) in epidermal cells, denaturing or cross-linking extracellular matrix components, resulting in reduced skin elasticity and inducing abnormal melanin pigment deposition, resulting in age spots and freckles. Mitochondria are the main source of ROS in the body, and it has been reported that increased ROS levels due to mitochondrial damage accelerate skin aging. Epidermis-specific MITOL knockout mice exhibit skin abnormalities, such as increased cell death in the epidermis, which has been suggested to be caused by increased ROS levels in epidermal keratinocytes (Non-Patent Document 3). Therefore, ingredients that promote MITOL production or inhibit MITOL reduction can prevent or improve skin aging by preventing increased ROS levels in epidermal cells.

The composition of the present invention containing Moutan Peony and taurine has the effect of inhibiting the decrease in MITOL or promoting MITOL production. This can prevent or improve symptoms of hair follicle aging, such as gray hair and hair loss, and symptoms of skin aging, such as a decrease in skin barrier function and age spots and wrinkles. It can also be used to prevent or improve symptoms of cardiac aging, such as decreased cardiac function, and symptoms of brain aging, such as Alzheimer's disease. Furthermore, it can be used as a reagent to promote MITOL production, and is ideally suited for use as a positive control when conducting material screening, etc.

The compositions of the present invention and their instructions may be labeled, for example, to be used to inhibit the decrease or promote the production of MITOL, to be used to prevent or improve hair follicle aging, to be used to prevent or improve gray hair, to be used to prevent or improve hair loss, to be used to prevent or improve skin aging, to be used to prevent or improve a decrease in skin barrier function, to be used to prevent or improve wrinkles, to be used to prevent or improve spots, or to be used for anti-aging. Here, "labeled" includes being labeled on the product itself, container, packaging, etc. containing the composition of the present invention; being labeled in documents such as instructions, package inserts, pamphlets, and other printed materials disclosing product information; and being labeled in various flyers and advertisements used for promotional purposes, including online advertising.

The present invention will be explained in more detail below using examples and test examples, but the present invention is not limited to these examples.

Test Example: Evaluation of the MITOL Production-Promoting Effect of Peony and Taurine. Human hair follicle keratinocytes (ScienCell Research Laboratories) were seeded in Humedia-KG2 medium (Kurashiki Boseki, supplemented with the attached supplements) onto a 24-well plate and cultured for several days in an incubator set at 37°C and 5% CO2. When the appropriate cell density was reached, the medium was replaced with medium containing the test substance and cultured for an additional 48 hours. After culture, the medium was removed, and cells were lysed by adding lysate buffer, and the cell lysate was collected. RNA was extracted from the cell lysate using the RNeasy Mini Kit (Qiagen) according to the attached protocol. Using this as a template, cDNA was synthesized by reverse transcription using Prime Script RT Master mix (Takara Bio). The mRNA expression levels of RPLP2 and MITOL were measured (SYBR Green method) from the synthesized cDNA using a real-time PCR system (Step One Plus, Thermo Fisher Scientific), and the expression level of MITOL was normalized by the expression level of the internal standard gene, RPLP2. The following primers were used: RPLP2: HA067804, MITOL: HA192576 (both Takara Bio). The measured mRNA expression levels were compared with those of a control group (medium not containing the test substance) to evaluate the effect of the test substance on promoting MITOL production.

<Test Results>
The results of evaluating Moutan Pea extract (extract extracted with a mixture of ethanol and water containing approximately 3% solids by mass) at various concentrations are shown in Figure 1. Moutan Pea exhibited the effect of promoting MITOL production, and this effect increased in a concentration-dependent manner.
Next, the results of using Moutan Pea in combination with taurine are shown in Figure 2. As with the results shown in Figure 1, under these test conditions, the MITOL production-promoting effect of Moutan Pea at a concentration of 0.1% by mass (approximately 0.003% by mass on a solid basis) was weak. However, when taurine was added, the MITOL production-promoting effect was synergistically enhanced. This indicates that taurine enhances the MITOL production-promoting effect of Moutan Pea.

<Production Example: Cleaning Composition>
A cleaning composition having a pH of 6.0 was obtained by dissolving 7 g of sodium tetradecene sulfonate solution, 3 g of sodium methyl taurate of coconut oil fatty acid, 3 g of coconut oil fatty acid amidopropyl betaine solution, 2 g of coconut oil fatty acid N-methylethanolamide, 1 g of polyoxyethylene sorbitan monolaurate, 1 g of polyoxypropylene coconut oil fatty acid monoisopropanolamide (1 P.O.), 0.3 g of O-[2-hydroxy-3-(trimethylammonio)propyl]hydroxyethylcellulose chloride, 0.1 g of dipotassium glycyrrhizinate, 0.1 g of salicylic acid, 0.1 g of 1-menthol, 0.5 g of methyl parahydroxybenzoate, 0.2 g of sodium benzoate, 0.05 g of Moutan Pea Extract, 0.03 g of taurine, 0.01 g of Coptis Rhizome Extract, 0.02 g of Phellodendron Bark Extract, an appropriate amount of fragrance, an appropriate amount of citric acid, an appropriate amount of sodium citrate, and purified water to a total amount of 100 g.

<Production example: hair composition>
A hair composition having a pH of 4.3 was obtained by adding 2 g of alkyltrimethylammonium chloride, 1.5 g of hydrogenated rapeseed oil alcohol, 2.5 g of cetanol, 2 g of liquid paraffin, 0.3 g of O-[2-hydroxy-3-(trimethylammonio)propyl]guar gum chloride, 3 g of concentrated glycerin, 3 g of 1,3-butylene glycol, 0.5 g of l-menthol, 0.1 g of dipotassium glycyrrhizinate, 0.1 g of salicylic acid, 0.1 g of methyl parahydroxybenzoate, 0.1 g of Moutan Pea Extract, 0.01 g of taurine, 0.05 g of Coptis Rhizome Extract, 0.05 g of Phellodendron Bark Extract, an appropriate amount of fragrance, an appropriate amount of citric acid, an appropriate amount of L-arginine, and purified water to a total amount of 100 g.

<Production Example: Hair Growth Composition>
A hair growth composition with a pH of 6.0 was obtained by adding 50 g of ethanol, 2 g of concentrated glycerin, 2 g of 1,3-butylene glycol, 0.5 g of polyoxyethylene hydrogenated castor oil, 1 g of l-menthol, 0.3 g of Peony extract, 0.05 g of taurine, 0.3 g of Coptis Rhizome extract, 0.3 g of Phellodendron Bark extract, an appropriate amount of fragrance, an appropriate amount of citric acid, an appropriate amount of sodium citrate, and purified water to a total volume of 100 g.

The composition of the present invention, which is characterized by containing Moutanga pisi and taurine, is expected to be effective in preventing or ameliorating, for example, hair follicle aging symptoms such as gray hair and hair loss, and skin aging symptoms such as decreased skin barrier function and age spots and wrinkles, by inhibiting MITOL reduction or promoting MITOL production. It can also be used to prevent or ameliorate cardiac aging symptoms such as decreased cardiac function, and brain aging symptoms such as Alzheimer's disease. Alternatively, it can be used as a reagent to promote MITOL production, and is preferably utilized as a positive control drug when conducting material screening, etc.

Claims (2)

A composition for preventing or improving gray hair, comprising Peony root extract and taurine, wherein the mass ratio of Peony root extract to taurine (Peony root extract (solid content): taurine) is 1:50 to 1:300. A composition for preventing or improving hair loss, comprising Peony Fruit and taurine, wherein the mass ratio of Peony Fruit to taurine (Peony Fruit (solid equivalent): taurine) is 1:50 to 1:300.