JP7682667B2 - Perlecan production promoter - Google Patents

Description

The present invention relates to a perlecan production promoter containing an extract of bellflower leaves.

Starting from the surface layer where the human body comes into contact with the outside air, the skin is made up of three layers: the epidermis, dermis, and subcutaneous tissue. The epidermis is made up of cells called keratinocytes, and from the deepest part close to the dermis, it is divided into the basal layer, spinous layer, granular layer, and stratum corneum. In the epidermis, keratinocytes are pushed up from the basal layer toward the stratum corneum by division, and are exfoliated from the surface layer as so-called dirt.

The dermis is divided into the papillary layer, which is close to the epidermis, and the reticular layer, which lies deeper. The papillary layer contains collagen fibers made up mainly of collagen and elastic fibers made up of elastin, as well as other extracellular matrix components and fibroblasts. The reticular layer is thicker than the papillary layer and makes up the majority of the dermis. The reticular layer also contains collagen fibers made up of collagen and elastic fibers made up of elastin, but they are thicker than the collagen fibers of the papillary layer and are more mature than the elastic fibers of the papillary layer.

The upper part of the papillary layer has papillae that protrude toward the epidermis. The epidermis layer fits between these papillae, and an uneven structure is formed between the epidermis and dermis, where the papillary layer of the dermis and the epidermis layer interlock. The epidermis and dermis are separated by a membrane structure called the epidermal basement membrane, and the papillae can be said to be a structure composed of the epidermis, dermis, and epidermal basement membrane.

Signal transmission and transport of waste products and nutrients between the dermis and epidermis are carried out through the papillae, and it is said that the interlocking of the dermis and epidermis has the effect of mitigating physical stimuli from the outside. It has also been reported that there is a relationship between the skin condition (stratum corneum water content, transepidermal water loss, stratum corneum cell area, skin color L ^* a ^* b ^* value) and the shape of the papillae (Patent Document 1). While the papillae have such an important function for the skin, it is known that the papillae become flattened due to aging and ultraviolet rays (Non-Patent Document 1), and the shape of the dermal papillae changes not only due to aging and ultraviolet rays, but also due to diseases. For example, it is said that papillae are excessively formed in psoriasis, and papillae are reduced or disappear in actinic keratosis (Non-Patent Document 2). As mentioned above, changes in papillae structure are closely related to changes in skin condition, so it is considered important to properly maintain the papillae shape in order to maintain normal skin condition or improve from abnormal conditions.

Various compositions have been proposed to improve skin conditions by improving papillae, including a cream containing vitamin C (Non-Patent Document 3) and a composition containing cape gooseberry fruit extract (Patent Document 2). In particular, Patent Document 2 discloses that the use of the composition improves wrinkles and skin gloss, creating a situation in which a papillae structure improver with high efficacy is expected. However, there are no materials other than those reported above that can be used as papillae structure improvers, and there is a demand for new papillae structure improvers that can be used industrially.

The basement membrane, one of the structures that make up the papillae, is a thin membrane-like structure that exists not only in skin but in all tissues, and exists between epithelial tissue and mesenchymal tissue, which are developmentally distinct, and functions to maintain a physically stable structure by connecting the two. Among them, the epidermal basement membrane exists between the epidermis and dermis of the skin, strongly connects the epidermis and dermis, and acts as a scaffold for epidermal cells, regulating the function of epidermal cells and maintaining the structure of the dermis. It is known that the main components of the epidermal basement membrane include laminin, collagen IV, VII, XVII, perlecan (also known as proteoglycan), and nidogen (Non-Patent Document 4). In the skin, the components of the epidermal basement membrane are produced by keratinocytes and fibroblasts that exist in the surrounding area.

Among the components that make up the epidermal basement membrane, laminin, collagen IV, VII, and XVII, which function to physically support the structure of tissue, are important molecules that are likely to contribute to changes and improvements in the papilla structure, i.e., the epidermal basement membrane structure. However, there have been no reports to date of improvement of the papilla structure by promoting the production of these molecules.
On the other hand, although perlecan is a basement membrane component like laminin, it merely controls cell proliferation and differentiation by supplying growth factors such as b-FGF and TGFβ to adjacent basal epidermal cells (Non-Patent Document 5) and does not have the function of physically supporting the structure of the epidermal basement membrane. Therefore, it is not thought to contribute to changes or improvement of the epidermal basement membrane structure, and it was even not expected that perlecan would contribute to changes or improvement of the papilla structure.

The Locust is also called Tendai Locust, and its roots have been used as a traditional Chinese medicine since ancient times. The rhizome, root bark, or extracts of the Locust have been reported to promote hair growth (Patent Document 3), to be antiallergic (Patent Document 4), and to inhibit melanin production (Patent Document 5). However, their effect on the papilla structure is not known at all. In addition, the alcohol extract of the leaves is known to have an antibacterial effect (Patent Document 6), but the skin-improving effect of the extract on the skin is not known at all. It has been disclosed that a skin topical preparation containing a large amount of an extract obtained by extracting dried and chopped Locust leaves at high temperature or for a long period of time has been shown to have a skin firming and wrinkle-improving effect (Patent Document 7), but the effect on perlecan and the papillae was not known. In addition, the extract obtained by extracting dried and chopped leaves at high temperature or for a long period of time gives the skin topical preparation a unique color and odor, adding factors that affect the user's preference, and also has the aspect of reducing the stability of the skin topical preparation, making it difficult to use industrially.

JP 2016-016277 A JP 2014-043400 A JP 2000-344632 A JP 2000-169383 A Japanese Patent Application Laid-Open No. H11-279069 JP 2001-097871 A JP 2002-363028 A

Kawasaki K, Yamanishi K, Yamada H, Int J Dermatol. 2015 54(3):295-301. Hiroshi Shimizu, New Skin Science, 2011, 38-42 Kirsten Sauermann, Soren Jaspers, Urte Koop, Horst Wenck. BMC Dermatol 2004 Sep 29;4(1):13. doi: 10.1186 471-5945-4-13 Dirk Breitkreutz, Isabell Coxholt, Kathrin Thiemann, Roswitha Nischt Biomed Res Int. 2013; 2013:179784. doi: 10.1155/2013/179784. Epub 2013 Mar 21. Eduardo M. Vidal 2013 The Basement Membrane Zone: Making the Connection

The objective of the present invention is to provide a perlecan production promoter and a papilla structure improver.

As a result of intensive research conducted by the inventors to solve the above-mentioned problems, they discovered that an extract of Uguisu Leaf has a remarkable effect on promoting perlecan production and improving the structure of the papillae, thereby solving the above-mentioned problems.

The present invention makes it possible to provide a perlecan production promoter and a papilla structure improver.

FIG. 1 is a graph showing the changes in perlecan gene expression levels in keratinocytes and fibroblasts due to aging and ultraviolet radiation. FIG. 2 is a graph showing the change in expression of perlecan-producing gene (mRNA) in keratinocytes and fibroblasts by the addition of an extract from Loquatica leaf. FIG. 3 shows images showing changes in perlecan protein expression in fibroblasts due to the addition of an extract from Loquatica leaf. FIG. 4 is an observation image showing the effect of improving the papilla structure by a composition containing an extract of Loquat leaf.

The locust (also known as Tendai locust) used in the present invention is Lindera strychnifolia, which belongs to the genus Strychnifolia and belongs to the family Lauraceae. The part used is the leaves, but other parts such as roots, bark, trunks, leaves, and flowers may be used together with the leaves as long as the effect is not hindered. The form of the leaves used for extraction is preferably one that has not been artificially crushed or shredded, although it is not a problem if they are slightly damaged during processes such as drying and transportation. Extracts obtained from locust leaves that have been artificially crushed or shredded have a strong color and smell, which may be a hindrance to practical use.

The extraction solvent for the aconite is not particularly limited, but for example, various suitable organic solvents are used to extract the aconite at low to high temperatures. Examples of the extraction solvent that can be used include one or more of the following: water; lower monohydric alcohols such as methyl alcohol and ethyl alcohol; liquid polyhydric alcohols such as glycerin, propylene glycol, and 1,3-butylene glycol; ketones such as acetone and methyl ethyl ketone; alkyl esters such as ethyl acetate; hydrocarbons such as benzene and hexane; ethers such as diethyl ether; and halogenated alkanes such as dichloromethane and chloroform. Among these, a 40% to 60% aqueous ethanol solution is preferred, and a 50% aqueous ethanol solution is particularly suitable.

The method for extracting the Loquat Leaf extract that can be used in the present invention is not particularly limited, but for example, when using dried leaves, it is preferable to use 1 to 1000 times, and especially 10 to 100 times, the amount of solvent by mass, and perform the extraction at 0°C or higher, especially 20°C to 40°C, for 1 hour or more, especially 1 to 7 days, and more preferably 1 to 3 days. Alternatively, the extraction may be performed by heating at 50 to 70°C for 1 to 3 hours. Extracts obtained by extraction at high temperatures exceeding 70°C or extraction for a long period of time exceeding 7 days have a strong color and odor, which may interfere with practical use.

The fringe leaf extract obtained under the above conditions may be used as the extracted solution, but it can also be used after being appropriately processed by filtration, concentration, powderization, decolorization, purification, etc., within the scope that does not impair the effects of the present invention.

When preparing a composition containing the perlecan production promoter and papilla structure improver of the present invention as described above, the amount of the loquat leaf extract may be appropriately adjusted according to the degree of the desired effect and is not particularly limited. For example, the amount is preferably 0.001 to 20.0% by mass, calculated as the evaporated dry matter, and is most preferably in the range of 0.01 to 5.0% by mass.

In the present invention, promotion of perlecan production includes promoting the perlecan gene (named Heparan Sulfate Proteoglycan 2 (HPGS2)) and protein in cells, as well as perlecan in a state in which a glycan is covalently bound.

In the present invention, the improvement of papilla structure refers to an improvement from changes in papillae caused by aging, ultraviolet rays, skin diseases, etc., from normal, and is a concept that includes not only changes in the number of papillae, but also improvements from deformation and flattening of papillae. Specifically, it refers to an improvement from changes in shape such as a decrease in the number of papillae, a change in the horizontal cross section from a normal shape close to a perfect circle or ellipse to a horizontal cross section that deviates from a perfect circle or ellipse, or an increase in the variation in the diameter in the short axis direction for each papilla, i.e., deformation, or a decrease in the size in the long axis direction, i.e., flattening of the papillae. Papillae can be observed with a confocal laser biomicroscope that can non-invasively observe the internal structure of the skin, and the number of papillae can be evaluated by counting the number of papillae present per observation area, flattening of the papillae can be evaluated by measuring the size in the long axis direction, and improvement from changes in shape can be evaluated by observing the size and shape in the short axis direction.

In addition, when preparing and using a composition containing the perlecan production promoter and papilla structure improver of the present invention, other ingredients such as oils and fats, waxes, hydrocarbon oils, ester oils, higher alcohols, silicone oils, UV absorbers, UV scattering agents, moisturizers, surfactants, water-soluble polymers, thickeners, powders, skin protective agents, whitening agents, wrinkle improving agents, anti-aging agents, plant extracts, preservatives, anti-inflammatory agents, pH adjusters, sequestering agents, antioxidants, stabilizers, fragrances, colorants, pigments, etc., can be appropriately blended as needed within the scope that does not impair the effects of the present invention.

The formulation of the perlecan production promoter and papilla structure improver of the present invention is not particularly limited. For example, the formulation may be a spray, liquid, gel, cream, solid, sheet, granule, tablet, or two-component formulation.

The present invention will be explained in more detail below with reference to examples, but the present invention is not limited to these examples. Furthermore, unless otherwise specified, the blending amounts are shown in mass %.

--Preparation of foxglove leaf extract--
50g of 50% ethanol (50 times the mass) was added to 1.0g of dried loquat leaves, and the mixture was extracted and dried at 50°C for 3 hours to obtain 2.5g of loquat leaf extract. 50g of 40% ethanol (50 times the mass) was added to 1.0g of dried loquat leaves, and the mixture was extracted and dried at 50°C for 3 hours to obtain 2.7g of loquat leaf extract. 50g of 60% ethanol (50 times the mass) was added to 1.0g of dried loquat leaves, and the mixture was extracted and dried at 50°C for 3 hours to obtain 2.2g of loquat leaf extract.

-Gene expression test-
<Confirmation of changes in perlecan gene expression due to aging and ultraviolet rays>
5.0 x 10 ⁴ cells/mL of human epidermal keratinocytes derived from a 23-year-old adult donor and human epidermal keratinocytes derived from a 62-year-old adult donor were dispersed in Humedia KG2 (KURABO) and 500 μL of each were seeded on a 24-well cell culture plate. After 72 hours of culture under conditions of 37°C and 5% (volume%) CO ₂ , total RNA extraction was performed. The medium in the well in which the cells that were not subjected to total RNA extraction were seeded was replaced with 300 μL of PBS (-), and 20 mJ of ultraviolet irradiation was performed. As a sample not exposed to ultraviolet light, a plate was wrapped in aluminum foil and treated in the same manner to prevent exposure to ultraviolet light. After further culturing for 3 hours under conditions of 37°C and 5% (volume%) CO ₂ , total RNA extraction was performed.

5.0 x 10 ⁴ cells/mL of human dermal fibroblasts derived from a 21-year-old adult donor and human dermal fibroblasts derived from a 69-year-old adult donor were dispersed in 10% FBS-containing DMEM (Gibco) and seeded in 500 μL each in a 24-well cell culture plate. After 72 hours of culture under conditions of 37 ° C and 5% (volume%) CO ₂ , total RNA extraction was performed. The medium in the well in which the cells that were not subjected to total RNA extraction were seeded was replaced with 300 μL of PBS (-), and 20 mJ of ultraviolet irradiation was performed. As a sample not exposed to ultraviolet light, a plate was wrapped in aluminum foil and treated in the same manner to prevent exposure to ultraviolet light. After further culturing for 3 hours under conditions of 37 ° C and 5% (volume%) CO ₂ , total RNA extraction was performed.

<Searching for substances that promote perlecan gene expression>
Human epidermal keratinocytes derived from newborn donors (5.0 x ¹⁰⁴ cells/mL) were dispersed in Humedia KG2 and 500 μL was seeded in a 24-well cell culture plate. After 24 hours of culture at ₃₇ °C and 5% (volume %) CO2, a medium diluted with Humedia KG2 so that the solid evaporation residue of 40%, 50%, and 60% ethanol extract of Loquat leaf was 100 ppm and 10 ppm, respectively, was added, and the culture was continued for 48 hours, after which total RNA was extracted.

Human dermal fibroblasts derived from newborn donors (5.0 x ¹⁰⁴ cells/mL) were dispersed in 10% FBS-containing DMEM and 500 μL of each were seeded in a 24-well cell culture plate. After 24 hours of culture at ₃₇ °C and 5% (volume %) CO2, a medium diluted with DMEM so that the solid evaporation residue of 40%, 50%, and 60% ethanol extracts of Loquat leaf was 100 ppm and 10 ppm was added, and the culture was continued for 48 hours, after which total RNA was extracted.

Prime Script RTP was performed on the total RNA extracted by the above method.
Reverse transcription was performed using CR KIT (TaKaRa) to synthesize cDNA. Using the obtained cDNA as a template, the expression levels of perlecan (HPSG2) and GAPDH were measured by real-time PCR (7500 Real Time PCR System, Applied Biosystems) using the following primers and enzymes.

The primers used were a sense primer for HPSG2 (5'-ATTTCCACGATGATGGCT-3'), an antisense primer (5'-ACCTCCAGCTCGATGGTC-3'), and a sense primer for GAPDH (glyceraldehyde 3-phosphate dehydrogenase: used as a housekeeping gene) (5'-ATTTCCACGATGATGGCT-3'), an antisense primer (5'-ACCTCCAGCTCGATGGTC-3'). SYBR Select Master Mix (Applied Biosystems) was used for the PCR reaction, and gene expression was analyzed by the comparative CT method. The results are shown in Figure 1.

Figure 1 shows that the expression level of perlecan (HPSC2) gene is lower in aged cells than in young cells in both fibroblasts and keratinocytes. It was also shown that the expression level of perlecan gene is reduced by exposure to ultraviolet light.

Figure 2 shows the results of an investigation into the effect of promoting perlecan gene expression. It was found that in both fibroblasts and keratinocytes, the group to which a 50% ethanol extract of loquat leaf was added had expression levels that were approximately 1.5-2.4 times higher than the control group. A similar increase in expression was also observed in the groups to which a 40% ethanol extract of loquat leaf was added and in the groups to which a 60% ethanol extract of loquat leaf was added. Since perlecan in the skin is known to be produced by fibroblasts and keratinocytes, it was confirmed that loquat leaf extract has the effect of promoting perlecan production in the skin.

<Confirmation of the effect of Uguisu Leaf Extract in promoting perlecan protein production>
Human dermal fibroblasts derived from newborn donors (5.0 x ¹⁰⁴ cells/mL) were dispersed in 10% FBS-containing DMEM and seeded in 500 μL per well of a 24-well cell culture plate. After 24 hours of culture at ₃₇ °C and 5% (volume%) CO2, a medium diluted with DMEM so that the solid evaporation residue of a 50% ethanol extract of Loquat leaf was 50 ppm was added, and the medium was cultured at 37°C and 5% (volume%) CO2 for ₂ weeks, and then immunostaining of perlecan protein was performed as follows. After fixing the cells with 4% PFA, a perlecan primary antibody (Anti-Heparan Sulfate Proteoglycan (Perlecan) Antibody, clone 5D7-2E4, Host Species Mouse (Merck)) was added and incubated at 4°C for 16 hours. A fluorescently labeled secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113) was added and incubated at room temperature for 45 minutes. Observation was performed using a fluorescence microscope (fluorescence microscope BZ-X700, Keyence, objective lens ×20, excitation wavelength 470 nm, fluorescence wavelength 525 nm, exposure time 1/2 second).

Figure 3 shows the results of an investigation into the effect of promoting perlecan protein expression. Areas with high perlecan protein production were stained, and it was found that the group in which a 50% ethanol extract of Loquat Leaf was added to the fibroblasts produced more perlecan than the control. It was confirmed that Loquat Leaf Extract not only promotes perlecan gene expression, but also the production of perlecan protein.

[Human use test of a composition containing an extract of crocus leaf]
A composition containing a 50% ethanol extract of Loquat leaves and a comparative composition shown in Table 1 were prepared, and the effect of improving the papilla structure and the effect of improving the skin condition were confirmed. Eight panelists (men aged 30 to 50 years) were asked to apply an appropriate amount of the composition to half of their face, once in the morning and once at night, for two months. The papilla was observed at the corners of the eyes and cheeks using a confocal laser biomicroscope (Vivascope 1500, Caliber I.D.). Wrinkles were measured by creating wrinkle replicas at the corners of the eyes using a silicone rubber replica agent SILFLO (Amic Group), and analyzing them using a three-dimensional image analyzer (PRIMOS, Canfield Scientific). The moisture content of the stratum corneum was measured at the cheeks using a SKICON 200-EX (I.B.S.). Skin color (L value: brightness) was measured on the cheek using a spectrophotometer CM-700d/600d (KONICA MINOLTA).

The papilla observation images were analyzed by obtaining a horizontal image of 1.0 mm x 1.0 mm approximately 25 μm from the beginning of observation of the papilla when observing from the skin surface toward the depths, and analyzing the papilla shape (number of papillae, deformation rate). The papilla deformation rate was calculated by counting the number of papillae in the image that had a cross-sectional shape that could be judged to be almost a perfect circle or ellipse (number of non-deformed papillae) and the number of papillae that had a cross-sectional shape that could be judged to be non-circular rather than a perfect circle or ellipse (number of deformed papillae), and the deformation rate of the papilla cross-sectional shape (papilla deformation rate) was calculated using the following formula 1.

The results of the analysis of the papillae images are shown in Table 2-3. An increase in the number of papillae and a decrease in the deformation rate were observed by using a composition containing Uguisu Leaf Extract. Images of a remarkable effect are shown in Figure 4. In both the cheek and eye corner images, it was observed that the number of papillae per unit area increased by using a composition containing Uguisu Leaf Extract. In addition, the number of deformed papillae as shown in the area surrounded by the white dotted line also decreased, and the number of papillae that were close to perfect circles increased, indicating that the papillae deformation rate also decreased. In addition, the effects of improving the skin condition were observed, including improvement of wrinkles at the eye corner, increase in the moisture content of the stratum corneum, and increase in the L value (brightness), which is the skin color, and the improvement of the skin condition thought to be due to the papillae structure. It was confirmed that excellent improvement in skin condition was observed due to the promotion of perlecan production and the effect of improving the papillae structure of Uguisu Leaf Extract. In addition, because the compositions used in the test did not have the distinctive color or odor associated with the loquat leaf extract, throughout the test period the panelists were unable to determine from the compositions which of the two compositions they were using contained the loquat leaf extract, confirming that no additional factors would affect the user's preferences. Furthermore, the loquat leaf extract had no effect whatsoever on the long-term stability of the composition.

<Judgment criteria>
- Number of nipples -
** (significant effect): The number of papillae after application is 120% or more compared to the number before application.
* (Effective): The number of papillae after application is 110% or more of the number before application. (Ineffective): The number of papillae after application is less than 110% of the number before application.

The results of the papillae count measurement are shown in Table 2.

<Judgment criteria>
- Nipple projection deformation rate -
** (Excellent effect): The rate of papilla deformation after application was less than 50% of the rate before application.
* (Effective): The papilla deformation rate after application is less than 70% of the papilla deformation rate before application - (Ineffective): The papilla deformation rate after application is 70% or more of the papilla deformation rate before application

The results of the nipple deformation rate measurement are shown in Table 3.

<Judgment criteria>
- Wrinkle area ratio -
* (Effective): The wrinkle area ratio after application is less than 80% of the wrinkle area ratio before application. (Ineffective): The wrinkle area ratio after application is 80% or more of the wrinkle area ratio before application.

The results of the wrinkle area ratio measurement are shown in Table 4.

<Judgment criteria>
- Moisture content of the stratum corneum -
* (Effective): The moisture content of the stratum corneum after application is 120% or more compared to the moisture content before application. (Ineffective): The moisture content of the stratum corneum after application is less than 20% compared to the moisture content before application.

The results of the measurement of the moisture content of the stratum corneum are shown in Table 5.

<Judgment criteria>
-Skin brightness-
* (Effective): L value after application is 110% or more of the L value before application started. (Ineffective): L value after application is less than 110% of the L value before application started.

The results of the brightness measurements are shown in Table 6.

Next, examples of the formulations of the agents of the present application containing the Loquat Leaf Extract of the present invention are shown, but the present invention is not limited thereto. The numerical values written after the component names indicate the blend amounts, and each formulation example is expressed as a total of 100% by mass. The Loquat Leaf Extract shown in each of the following formulation examples was prepared in Example 1, and its blend amount is shown as mass % converted to evaporation residue. The effects of the present application were also confirmed in each formulation example.
(Formulation Example 1) When used as an emulsion composition (% by mass)
a) Beeswax: 2.0
b) Stearyl alcohol...5.0
c) Stearic acid...8.0
d) Squalane...10.0
e) Self-emulsifying glyceryl monostearate...3.0
f) Polyoxyethylene cetyl ether (20 E.O.)...1.0
g) 50% ethanol extract of foxtail leaf...0.01
h) 1,3-butylene glycol...5.0
i) Potassium hydroxide...0.3
j) Preservatives and antioxidants... appropriate amount k) Purified water... balance total... 100
Manufacturing method: Heat and dissolve a) through f) and keep at 80°C. Heat and dissolve h) through k) and keep at 80°C, add to a) through f) and emulsify, then cool to 40°C with stirring. Then add g) and stir to dissolve uniformly.

(Formulation Example 2) When used as a liquid composition (% by mass)
a) 60% ethanol extract of cinnamon leaf...0.01
b) Glycerin...5.0
c) Polyoxyethylene sorbitan monolaurate (20 E.O.) ... 1.0
d) Ethanol...6.0
e) Fragrance... appropriate amount f) Preservatives and antioxidants... appropriate amount g) Purified water... balance Total... 100
Manufacturing method: Mix a) to g) and dissolve uniformly.

(Formulation Example 3) When used as a gel composition (% by mass)
a) 40% ethanol extract of foxtail leaf...0.01
b) Carboxyvinyl polymer...0.5
c) Sodium hydroxide...0.05
d) Methyl paraoxybenzoate...0.1
e) Cyclohexane-1,4-dicarboxylic acid bisethoxydiglycol ... 0.5
f) (Eicosanedioic acid/tetradecanedioic acid) polyglyceryl-10...0.5
g) PEG/PPG/Polybutylene Glycol-8/5/3 Glycerin...0.5
h) Polyoxyethylene (60 E.O.) hydrogenated castor oil...0.1
i) Fragrance... appropriate amount j) Preservatives and antioxidants... appropriate amount k) Purified water... balance Total... 100
Manufacturing method: Disperse b) in a part of k), then add c). Then mix a) to k) and dissolve uniformly.

(Formulation Example 4) When used as a powder composition (% by mass)
a) 50% ethanol extract of foxtail leaf...20.0
b) Cyclohexane-1,4-dicarboxylic acid bisethoxydiglycol ... 0.2
c) (Eicosanedioic acid/tetradecanedioic acid) polyglyceryl-10...0.2
d) PEG/PPG/Polybutylene Glycol-8/5/3 Glycerin...0.2
e) Sodium bicarbonate...50.0
f) Sodium sulfate... remainder g) Fragrance... appropriate amount h) Preservatives and antioxidants... appropriate amount Total... 100
Preparation method: Mix a) to h) uniformly.

Claims (4)

A perlecan production promoter containing an extract of Loquats leaves extracted from Loquats at 70°C or less for 3 days or less without artificial crushing or shredding . The papilla structure improving agent contains an extract of Loquat leaves extracted from Loquat at 70°C or less for 3 days or less without artificial crushing or shredding . Extracted from Uguisu Rhizome at 70°C or below for 3 days or less without artificial crushing or shredding for industrial use. Use of an extract of Uguisu Rhizome leaf or a composition containing an extract of Uguisu Rhizome leaf to promote perlecan production (excluding use in treating humans) . Extracted from alfalfa at 70℃ or less for 3 days without artificial crushing or shredding for industrial use.
Use of an extract of Uguisudachi leaf or a composition containing an extract of Uguisudachi leaf for improving papilla structure (excluding for therapeutic use in humans) .