Pearl Kan production promoter

Abstract

To provide a perlecan production promoter and a papilla structure improver.SOLUTION: The inventors have performed extensive and intensive study to discover that extract from leaves of Lindera strychnifolia is significantly effective in perlecan production promotion and papilla structure improvement.EFFECT OF THE INVENTION: This invention can provide a perlecan production promoter and a papilla structure improver.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a pearl kan production promoter containing an extract of Lindera aggregata leaves.

Background Art

[0002] The skin is composed of three layers: the epidermis, the dermis, and the subcutaneous tissue, in order from the surface layer in contact with the outside air. The epidermis consists of cells called keratinocytes, and is classified into the basal layer, the spinous layer, the granular layer, and the stratum corneum from the deep part near the dermis. In the epidermis, keratinocytes are pushed upward from the basal layer toward the stratum corneum by division, and are peeled off as so-called dirt from the surface layer in order.

[0003] The dermis is classified into the papillary layer near the epidermis and the reticular layer existing deeper than that. The papillary layer mainly includes collagen fibers composed of collagen, elastic fibers composed of elastin, other extracellular matrix components, and fibroblasts. The reticular layer is thicker than the papillary layer and occupies most of the dermis. The reticular layer also includes collagen fibers composed of collagen and elastic fibers composed of elastin, but is thicker than the collagen fibers of the papillary layer and more mature than the elastic fibers of the papillary layer.

[0004] Papillae protruding toward the epidermis are formed in the upper part of the papillary layer. The epidermal layer enters between these papillae, and an uneven structure in which the papillary layer of the dermis layer and the epidermal layer are engaged with each other is formed between the epidermis and the dermis. The space between the epidermal layer and the dermis layer is separated by a membrane structure called the epidermal basement membrane, and it can be said that the papilla is a structure composed of the epidermal layer, the dermis layer, and the epidermal basement membrane.

[0005] Between the dermis and the epidermis, signal transmission and substance transport such as waste products and nutrients are performed through the papillae, and it is said that the engagement between the dermis and the epidermis has a relaxing effect on physical stimuli from the outside. In addition, skin conditions (stratum corneum water content, transepidermal water loss, stratum corneum cell area, skin color L * a * b *It has also been reported that there is a relationship with the value) and the shape of the papillae (Patent Document 1). While the papillae have such important functions for the skin, it is known that the papillae are flattened not only by aging and ultraviolet rays (Non-Patent Document 1), but also by diseases in addition to aging and ultraviolet rays, and there are shape changes in the dermal papillae due to diseases. For example, it is said that in psoriasis, papillae are excessively formed, and in actinic keratosis, papillae are reduced or disappear (Non-Patent Document 2). As described above, since the change in the papilla structure is closely related to the change in the skin state, it is considered important to appropriately maintain the papilla shape in order to maintain the skin state normally or to improve from the state where abnormalities occur.

[0006] So far, various compositions for improving the skin state by improving the papillae have been proposed. For example, a cream containing vitamin C (Non-Patent Document 3), a composition containing an extract of Physalis alkekengi fruits, etc. are known (Patent Document 2). In particular, Patent Document 2 discloses that the use of the composition exerts an effect of improving wrinkles and skin gloss, and a situation has arisen where a papilla structure improver having a high effect is expected. However, materials that can be used as papilla structure improvers are not available other than the above reports, and there has been a demand for a new industrially applicable papilla structure improver.

[0007] The basement membrane, which is one of the structures constituting the papillae, is a thin membranous structure. It exists not only in the skin but also in all tissues, and exists between epithelial tissues and mesenchymal tissues that are different in embryology and has the function of maintaining a physically stable structure by binding the two. Among them, the epidermal basement membrane exists between the epidermis and the dermis of the skin, strongly binds the epidermis and the dermis, and serves as a scaffold for epidermal cells to regulate the function of epidermal cells and maintain 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), nidogen, etc. (Non-Patent Document 4). In the skin, epidermal basement membrane components are produced by keratinocytes and fibroblasts existing around it.

[0008] Among the components that make up the epidermal basement membrane, laminin, collagen IV, VII, and XVII, which have the function of physically supporting the tissue structure, are important molecules as those that are likely to contribute to the change or improvement of the papillary structure, that is, the epidermal basement membrane structure. However, there has been no report on the improvement of the papillary structure by promoting the production of these molecules. On the other hand, perlecan, although it is a basement membrane component like laminin, etc., only 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 considered to contribute to the change or improvement of the epidermal basement membrane structure, and furthermore, it has not been assumed that perlecan contributes to the change or improvement of the papillary structure.

[0009] Lindera aggregata is also called Lindera strychnifolia. Its root has been used as a traditional Chinese medicine since ancient times. In addition, regarding the rhizome, root bark, or extract of Lindera aggregata, various effects such as hair growth (Patent Document 3), anti-allergy (Patent Document 4), and melanin production inhibition (Patent Document 5) have been reported, but its effect on the papillary structure is completely unknown. Also, the alcohol extract of its leaves is known to have antibacterial activity (Patent Document 6), but its improvement effect on the skin is completely unknown. It is disclosed that a skin external preparation containing a large amount of an extract obtained by high-temperature or long-term extraction of dried and shredded Lindera aggregata leaves has an effect of improving skin firmness and wrinkles (Patent Document 7), but its effect on perlecan and its effect on the papilla were unknown. In addition, the extract obtained by high-temperature or long-term extraction of dried and shredded leaves has a specific color and odor for skin external preparations, which adds elements that affect the user's preference, and also has the aspect of reducing the stability of skin external preparations, making it difficult for industrial use.

Prior Art Documents

Patent Documents

[0010]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Patent Document 6

Patent Document 7

Non-Patent Document

[0011]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Summary of the Invention

Problems to be Solved by the Invention

[0012] An object of the present invention is to provide a promoter for pearl can production and an agent for improving papillary protrusion structure.

Means for Solving the Problems

[0013] As a result of intensive studies to solve the above problems, the present inventors have found that the extract of Lindera aggregata leaves has a remarkable effect on promoting pearl can production and improving papillary protrusion structure, and have solved the above problems.

Effects of the Invention

[0014] According to the present invention, it is possible to provide a promoter for pearl can production and an agent for improving papillary protrusion structure.

Brief Description of the Drawings

[0015]

Figure 1

Figure 2

Figure 3

Figure 4

Modes for Carrying Out the Invention

[0016] The Lindera strychnifolia used in the present invention (also known as Lindera aggregata var. sutchuenensis) belongs to the genus Lindera of the Lauraceae family. The part used is the leaf, but as long as the effect is not inhibited, it can be used in a state where other parts such as roots, bark, trunk, leaves, and flowers are mixed with the leaves. The form for extraction may have some damage in the processes such as drying and transportation, but it is preferably not artificially pulverized or shredded. The extract obtained from artificially pulverized or shredded Lindera strychnifolia leaves has a strong color and odor, which may affect actual use.

[0017] The extraction solvent of Lindera strychnifolia is not particularly limited. For example, various suitable organic solvents can be used for extraction from low temperature to high temperature. As the extraction solvent, for example, 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; halogenated alkanes such as dichloromethane and chloroform, etc., one or more of them can be used. Among them, a 40% - 60% ethanol aqueous solution is preferred, and a 50% ethanol aqueous solution is particularly preferred.

[0018] The extraction method of the Lindera strychnifolia leaf extract that can be used in the present invention is not particularly limited. For example, when using dried ones, a solvent in an amount of 1 - 1000 times, particularly 10 - 100 times the mass ratio is used, and the extraction is preferably carried out at 0°C or above, particularly at 20°C - 40°C for 1 hour or more, particularly for 1 - 7 days, more preferably for 1 - 3 days. Also, heating extraction can be carried out at 50 - 70°C for 1 - 3 hours. The extracts obtained by extraction at a high temperature exceeding 70°C and extraction for a long period exceeding 7 days have a strong color and odor, which may affect actual use.

[0019] The Lindera strychnifolia leaf extract obtained under the above conditions can be used as the extracted solution, but it can also be used after appropriate treatments such as filtration, concentration, powdering, decolorization, and purification within a range that does not impair the effects of the present invention.

[0020] When preparing a composition containing the pearl can production promoter and the papilla structure improver of the present invention as described above, the blending amount of the Lindera aggregata leaf extract may be appropriately adjusted according to the degree of the desired effect and is not particularly limited. For example, it is preferably 0.001 to 20.0% by mass in terms of the dry matter by evaporation, and particularly preferably in the range of 0.01 to 5.0% by mass.

[0021] The promotion of pearl can production in the present invention includes promoting heparan sulfate proteoglycan 2 (HPGS2), a protein, and a pearl can in a state where a sugar chain is covalently bonded to the pearl can gene of cells.

[0022] The improvement of the papilla structure in the present invention is an improvement from the changes from normal due to aging, ultraviolet rays, skin diseases, etc. of the papilla, and is a concept that includes improvement from deformation and flattening of the papilla in addition to changes in the number of papilla. Specifically, when the number of papilla decreases, the direction perpendicular to the epidermal basement membrane is the major axis direction, and the direction intersecting horizontally is the minor axis direction, for the horizontal cross-section, the normal shape is close to a perfect circle or an ellipse, whereas the horizontal cross-section moves away from a perfect circle or an ellipse, or the variation in the minor axis diameter of each papilla becomes large, so-called deformation, and also refers to improvement from shape changes such as the size in the major axis direction becoming short, so-called flattening of the papilla. The papilla can be observed with a confocal laser scanning microscope that can non-invasively observe the internal structure of the skin, and the improvement from shape changes can be evaluated by counting the number of papilla present per observation area to measure the number of papilla, measuring the size in the major axis direction to observe the flattening of the papilla, and observing the size / shape in the minor axis direction.

[0023] When preparing and using a composition containing the pearl kan production promoter and papillary structure improver of the present invention, other components, such as fats and oils, waxes, hydrocarbon oils, ester oils, higher alcohols, silicone oils, ultraviolet absorbers, ultraviolet scattering agents, moisturizers, surfactants, water-soluble polymers, thickeners, powders, skin protectants, whitening agents, wrinkle improvers, anti-aging agents, plant extracts, preservatives, anti-inflammatory agents, pH adjusters, sequestering agents, antioxidants, stabilizers, fragrances, pigments, dyes, etc., can be appropriately blended as needed within the range that does not impair the effects of the present invention.

[0024] The dosage forms of the pearl kan production promoter and papillary structure improver of the present invention are not particularly limited. For example, forms such as spray agents, liquids, gels, creams, solids, sheets, granules, tablets, or two-component types are not particularly restricted.

Examples

[0025] Hereinafter, the present invention will be further specifically described by way of examples, but the present invention is not limited by these examples. Also, unless otherwise specified, the blending amounts are shown in mass %.

[0026] - Preparation of Lindera aggregata leaf extract - To 1.0 g of the dried Lindera aggregata leaves, 50 g of 50-fold mass of 50% ethanol was added, and extraction and drying were carried out at 50 °C for 3 hours to obtain 2.5 g of the Lindera aggregata leaf extract. To 1.0 g of the dried Lindera aggregata leaves, 50 g of 50-fold mass of 40% ethanol was added, and extraction and drying were carried out at 50 °C for 3 hours to obtain 2.7 g of the Lindera aggregata leaf extract. To 1.0 g of the dried Lindera aggregata leaves, 50 g of 50-fold mass of 60% ethanol was added, and extraction and drying were carried out at 50 °C for 3 hours to obtain 2.2 g of the Lindera aggregata leaf extract.

[0027] - Gene expression test - <Confirmation of changes in pearl kan gene expression levels due to aging and ultraviolet rays> 5.0×10 human epidermal keratinocytes derived from 23-year-old adult donors and human epidermal keratinocytes derived from 62-year-old adult donors 4Dispersed at Cells / mL in Humedia KG2 (KURABO) and seeded 500 μL each into a 24-well cell culture plate. After culturing for 72 hours under the conditions of 37 °C and 5% (v / v) CO 2 After culturing under 2 conditions for 72 hours, total RNA extraction was performed. The medium in the wells seeded with cells without total RNA extraction was replaced with 300 μL of PBS(-), and ultraviolet irradiation at 20 mJ was performed. As a non-ultraviolet-irradiated sample, a sample treated in the same manner by wrapping the plate with aluminum foil so that it was not irradiated with ultraviolet light was also prepared. Further, after culturing for 3 hours under the conditions of 37 °C and 5% (v / v) CO 2 Total RNA extraction was performed after culturing under 2 conditions for 3 hours.

[0028] Human dermal fibroblasts derived from a 21-year-old adult donor and human dermal fibroblasts derived from a 69-year-old adult donor 5.0×10 4 Dispersed at Cells / mL in DMEM containing 10% FBS (Gibco) and seeded 500 μL each into a 24-well cell culture plate. After culturing for 72 hours under the conditions of 37 °C and 5% (v / v) CO 2 After culturing for 72 hours under 2 conditions, total RNA extraction was performed. The medium in the wells seeded with cells without total RNA extraction was replaced with 300 μL of PBS(-), and ultraviolet irradiation at 20 mJ was performed. As a non-ultraviolet-irradiated sample, a sample treated in the same manner by wrapping the plate with aluminum foil so that it was not irradiated with ultraviolet light was also prepared. Further, after culturing for 3 hours under the conditions of 37 °C and 5% (v / v) CO 2 Total RNA extraction was performed after culturing under 2 conditions for 3 hours.

[0029] <Search for substances that promote the expression level of the Pearlkan gene> Human epidermal keratinocytes derived from a neonatal donor 5.0×10 4 Dispersed at Cells / mL in Humedia KG2 and seeded 500 μL each into a 24-well cell culture plate. After culturing for 24 hours under the conditions of 37 °C and 5% (v / v) CO 2 After culturing for 48 hours after adding a medium diluted with Humedia KG2 so that the solid evaporation residues of the 40%, 50%, and 60% ethanol extracts of Lindera aggregata leaves were 100 ppm and 10 ppm, total RNA extraction was performed.

[0030] Neonatal donor-derived human dermal fibroblasts 5.0×10 4 Cells / mL were dispersed in DMEM containing 10% FBS and seeded at 500 μL per well in a 24-well cell culture plate. After culturing at 37 °C under 5% (v / v) CO 2 conditions for 24 hours, media diluted with DMEM were added such that the solid evaporation residues of 40%, 50%, and 60% ethanol extracts of Lindera aggregata leaves were 100 ppm and 10 ppm, respectively. After culturing for 48 hours, total RNA was extracted.

[0031] For the total RNA extracted by the above method, reverse transcription was performed using Prime Script RT P CR KIT (TaKaRa) to synthesize cDNA. Using the obtained cDNA as a template, the expression levels of Perlcan (HPSG2) and GAPDH were measured by real-time PCR (7500 Real Time PCR System, Applied Biosystems) using the following primers and enzymes.

[0032] The primers used were the sense primer for HPSG2 (5´―ATTTCCACGATGATGGCT―3´), the antisense primer (5´―ACCTCCAGCTCGATGGTC―3´), the sense primer for GAPDH (glyceraldehyde 3-phosphate dehydrogenase, used as a housekeeping gene) (5´―ATTTCCACGATGATGGCT―3´), and the antisense primer (5´―ACCTCCAGCTCGATGGTC―3´). SYBR Select Master Mix (Applied Biosystems) was used for the PCR reaction, and gene expression analysis was performed by the comparative CT method. The results are shown in Figure 1.

[0033] From Figure 1, it was found that the expression level of the Perlcan (HPSC2) gene was lower in senescent cells than in young cells in both fibroblasts and keratinocytes. Also, it was shown that the expression level of the Perlcan gene decreased by ultraviolet irradiation.

[0034] Figure 2 shows the results of investigating the effect of promoting the expression level of the perlecan gene. In both the fibroblast and keratinocyte groups supplemented with 50% ethanol extract of Lindera aggregata leaves, the expression level was found to be about 1.5 - 2.4 times that of the control group. Similar increases in the expression level were also observed in the groups supplemented with 40% ethanol extract of Lindera aggregata leaves and 60% ethanol extract of Lindera aggregata leaves. Since it is known that perlecan in the skin is produced by fibroblasts and keratinocytes, it was confirmed that the Lindera aggregata leaf extract has an effect of promoting the production of perlecan in the skin.

Example

[0035] <Confirmation of the effect of Lindera aggregata leaf extract on promoting the production of perlecan protein> 5.0×10 neonatal donor-derived human dermal fibroblasts 4 Cells / mL were dispersed in DMEM containing 10% FBS and seeded at 500 μL per well in a 24-well cell culture plate. After culturing at 37 °C and 5% (v / v) CO 2 for 24 hours under conditions, a medium diluted with DMEM so that the solid evaporation residue of the 50% ethanol extract of Lindera aggregata leaves was 50 ppm was added, and then cultured at 37 °C and 5% (v / v) CO 2 for 2 weeks under conditions. Immunostaining of perlecan protein was performed according to the following procedure. 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 reacted at 4 °C for 16 hours. A fluorescently labeled secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor(R) 488) (ab150113) was added and reacted at room temperature for 45 minutes. Observation was carried out with a fluorescence microscope (fluorescence microscope BZ-X700, Keyence, objective lens ×20 magnification, excitation wavelength 470 nm, fluorescence wavelength 525 nm, exposure time 1 / 2 second).

[0036] Figure 3 shows the observation results of investigating the effect of promoting the expression level of pearl kantan protein. The areas with a high production amount of pearl kantan protein are stained, and it was found that in the group with the addition of 50% ethanol extract of Lindera aggregata leaves to fibroblasts, more pearls are produced compared to the control. It was also confirmed that the Lindera aggregata leaf extract has an effect of promoting not only the expression of the pearl kan gene but also the production of pearl kantan protein.

Example

[0037] [Human practical use test of the composition containing Lindera aggregata leaf extract] The composition containing 50% ethanol extract of Lindera aggregata leaves in [Table 1] and the composition of the comparative example were prepared, and the effect of improving the papillary structure and the effect of improving the skin condition were confirmed. Eight panelists (men aged 30 to 50) were asked to apply an appropriate amount of the composition to half of their face once in the morning and once in the evening for 2 months. The papillae were observed using a confocal laser scanning microscope (Vivascope 1500, Caliber I.D.) on the outer canthus and cheek. The measurement of wrinkles was performed by creating a wrinkle replica of the outer canthus using a silicone rubber replica agent SILFLO (Amic Group) and analyzing it using a three-dimensional image analyzer (PRIMOS, Canfield Scientific). The measurement of the water content of the stratum corneum was performed on the cheek using SKICON 200-EX (I.B.S.). The measurement of skin color (L value: lightness) was performed on the cheek using a spectrocolorimeter CM-700d / 600d (KONICA MINOLTA).

[0038]

Table 1

[0039] Analysis of the papilla observation images was performed as follows: When observing from the skin surface in the deep direction, a horizontal image at a position approximately 25 μm from the start of papilla observation was obtained as a 1.0 mm × 1.0 mm image, and the papilla shape (number of papillae, deformation rate) was analyzed. The papilla deformation rate was calculated by counting the number of papillae with a cross-sectional shape that can be determined as almost circular or elliptical in the image (number of non-deformed papillae) and the number of papillae with a cross-sectional shape that can be determined as non-circular rather than circular or elliptical (number of deformed papillae), and then obtaining the deformation rate of the papilla cross-sectional shape (papilla deformation rate) from the following formula 1.

[0040]

Number

[0041] The analysis results of the papilla images are shown in Table 2-3. By using the composition containing the extract of Lindera aggregata leaves, an increase in the number of papillae and a decrease in the deformation rate were observed. The observation images of the significant efficacy examples are shown in Figure 4. In both the images of the cheek and the outer corner of the eye, it was observed that the number of papillae per unit area increased by using the composition containing the extract of Lindera aggregata leaves. Also, as shown in the part surrounded by the white dotted line, the number of deformed papillae decreased, and the number of papillae close to a perfect circle increased. Therefore, it was found that the papilla deformation rate also decreased. In addition, as an effect of improving the skin condition, an improvement in the wrinkles at the outer corner of the eye, an increase in the amount of horny layer moisture, and an increase in the L value (lightness) of the skin color were observed, and an improvement in the skin condition considered to be due to the papilla structure was recognized. It was confirmed that excellent improvement in the skin condition was recognized due to the promotion of pearl can production by the extract of Lindera aggregata leaves and the effect of improving the papilla structure. Also, since the compositions used in the test did not have a specific color or odor derived from the extract of Lindera aggregata leaves, the panelists could not determine which of the two compositions during use contained the extract of Lindera aggregata leaves throughout the test period, and it was confirmed that no factor affecting the user's preference was added. Furthermore, the extract of Lindera aggregata leaves had no effect on the temporal stability of the composition.

[0042] <Judgment Criteria> ―Number of Papillae― **(Significant effect): The number of papillae after application is 120% or more of the number of papillae before application *(Effective): The number of papillae after application is 110% or more of the number of papillae before application ― (Ineffective): The number of papillae after application is less than 110% of the number of papillae before application Table 2 shows the determination results of the number of papillae measurements

[0043]

Table 2

[0044] <Judgment criteria> ― Papilla deformation rate― **(Significant effect): The papilla deformation rate after application is less than 50% of the papilla deformation 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 Table 3 shows the determination results of the papilla deformation rate measurements

[0045]

Table 3

[0046] <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 Table 4 shows the determination results of the wrinkle area ratio measurements

[0047]

Table 4

[0048] <Judgment Criteria> - Stratum corneum moisture content - *(Effective): The stratum corneum moisture content after application is 120% or more of that before application. - (Invalid): The stratum corneum moisture content after application is less than 120% of that before application. The judgment results of the stratum corneum moisture content measurement are shown in Table 5.

[0049]

Table 5

[0050] <Judgment Criteria> - Skin lightness - *(Effective): The L value after application is 110% or more of the L value before application. - (Invalid): The L value after application is less than 110% of the L value before application. The results of the lightness measurement are shown in Table 6.

[0051]

Table 6

[0052] Next, examples of the formulations of each agent of the present application containing the extract of Lindera aggregata leaves of the present invention are shown, but the present invention is not limited thereto. The numerical values described after the component names indicate the blending amounts, and each formulation example is expressed as a total of 100% by mass. The extract of Lindera aggregata leaves shown in each of the following formulation examples was prepared in Example 1, and its blending amount was shown as mass% converted to the evaporation residue. In addition, the effects of the present application were confirmed in each formulation example. (Formulation Example 1) When used as an emulsified composition (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 Lindera aggregata leaves ··· 0.01 h) 1,3-Butylene glycol ··· 5.0 i) Potassium hydroxide ··· 0.3 j) Preservative · Antioxidant ··· appropriate amount k) Purified water ··· the balance Total ··· 100 Manufacturing method a) to f) are heated and dissolved, and maintained at 80 °C. h) to k) are heated and dissolved, maintained at 80 °C, added to a) to f) and emulsified, and cooled with stirring to 40 °C. Then, g) is added and stirred until uniformly dissolved.

[0053] (Formulation Example 2) When used as a liquid composition (mass%) a) 60% ethanol extract of Lindera aggregata leaves ··· 0.01 b) Glycerin ··· 5.0 c) Polyoxyethylene sorbitan monolaurate (20 E.O.) ··· 1.0 d) Ethanol ··· 6.0 e) Fragrance ··· appropriate amount f) Preservative · Antioxidant ··· appropriate amount g) Purified water ··· the balance Total ··· 100 Manufacturing method a) to g) are mixed and uniformly dissolved.

[0054] (Formulation Example 3) When used as a gel composition (mass%) a) 40% ethanol extract of Lindera aggregata leaves ··· 0.01 b) Carboxyvinyl polymer ··· 0.5 c) Sodium hydroxide ··· 0.05 d) Methyl paraoxybenzoate ··· 0.1 e) Bis(ethoxydiglycol) cyclohexane-1,4-dicarboxylate ··· 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) Preservative · Antioxidant ··· Appropriate amount k) Purified water ··· The balance Total ··· 100 Manufacturing method Disperse b) in part of k), then add c). After that, mix a) to k) and dissolve them uniformly.

[0055] (Formulation Example 4) When used as a powdery composition (mass%) a) Lindera aggregata leaf 50% ethanol extract ··· 20.0 b) Bis(ethoxydiglycol) cyclohexane - 1,4 - dicarboxylate ··· 0.2 c) (Eicosanedioic acid / Tetradecanedioic acid) Polyglyceryl - 10 ··· 0.2 d) PEG / PPG / Polybutylene Glycol - 8 / 5 / 3 Glycerin ··· 0.2 e) Sodium hydrogen carbonate ··· 50.0 f) Sodium sulfate ··· The balance g) Fragrance ··· Appropriate amount h) Preservative · Antioxidant ··· Appropriate amount Total ··· 100 Manufacturing method Mix a) to h) uniformly.

Claims

**Claim 1**: A perlecan production promoter containing an extract of Lindera aggregata leaves extracted from Lindera aggregata at 70°C or lower within 3 days without artificial pulverization or shredding. **Claim 2**: A papillary structure improver containing an extract of Lindera aggregata leaves extracted from Lindera aggregata at 70°C or lower within 3 days without artificial pulverization or shredding. **Claim 3**: For industrial use, without artificial pulverization or shredding, extracted from Lindera aggregata at 70°C or lower within 3 days Use of an extract of Lindera aggregata leaves or a composition containing an extract of Lindera aggregata leaves for promoting perlecan production (except for use in treating humans). **Claim 4**: For industrial use, without artificial pulverization or shredding, extracted from Lindera aggregata at 70°C or lower within 3 days, Use of an extract of Lindera aggregata leaves or a composition containing an extract of Lindera aggregata leaves for improving papillary structure (except for use in treating humans).

Images