JP7456720B2 - Ceramide production promoter - Google Patents

Description

TECHNICAL FIELD The present invention relates to a ceramide production promoter containing as an active ingredient one or two selected from the group consisting of purple forsythia and lavender.

Ceramide in the skin is biosynthesized by forming a sphingosine skeleton in epidermal keratinocytes using the amino acids L-serine and palmitic acid as starting materials, and then through various enzymatic reactions (Non-Patent Document 1). Ceramide synthesized in epidermal keratinocytes is temporarily stored in the form of glucosylceramide or sphingomyelin, and then is excreted outside the cell and becomes ceramide again under the action of various enzymes and functions as an intercellular lipid. (Patent Document 1).

The metabolism of ceramide may be impaired by aging, exposure to ultraviolet light, dry skin, rough skin, atopic dermatitis, senile xerosis, psoriasis, and the like. As a result, it has been reported that the amount of ceramide in the stratum corneum decreases, causing, for example, a decrease in the skin's moisturizing and barrier functions. Recently, therefore, active research has been conducted on the production of substances aimed at promoting the production of epidermal ceramide in the stratum corneum of the skin. Examples of such ceramide production promoters include canna (Patent Document 2), eucalyptus (Patent Document 3), genkwanin (Patent Document 4), cnidium officinalis extract (Patent Document 5), sphingoids (Patent Document 6), and rhubarb (Patent Document 7).

Furthermore, in recent years, it has been reported that acylceramide is decreasing in skin diseases such as atopic dermatitis and xeroderma (Non-Patent Document 2). Furthermore, it has been reported that the water retention capacity of the epidermis is drastically reduced, and that mice engineered to be unable to produce only acylceramide develop significant skin abnormalities after birth and die due to water loss from the skin. (Non-patent document 3).
Thus, it is expected that improvement and normalization of epidermal barrier function will be promoted by promoting the production of ceramides, particularly acylceramides. However, with regard to substances that promote ceramide production, particularly acylceramide, that can be used in external skin preparations and foods and drinks, the existence of sufficient variations has not yet been confirmed.

"Cosmetics Encyclopedia" edited by the Japanese Society of Cosmetic Engineers, Maruzen Co., Ltd., December 15, 2003, p.563-564 J. Invest. Dermatol., 130, 2511-2514 (2010). Nature Communication (2017) DOI: 10.1038/ncomms14609

JP 2015-24993 Publication JP 2017-88539 Publication Japanese Patent Application Publication No. 2000-169359 Japanese Patent Application Publication No. 2004-161648 Japanese Patent Application Publication No. 2010-150237 Japanese Patent Application Publication No. 2012-92032 JP2014-74074

The present invention restores the ceramide production function that the body itself has, especially the acyl ceramide production function, which reduces the effects of aging, UV exposure, dry skin, rough skin, atopic dermatitis, senile xeroderma, psoriasis, etc. To provide a ceramide production promoter capable of restoring ceramide metabolism in the stratum corneum and increasing the amount of ceramide, thereby restoring skin having excellent moisturizing and barrier functions.

The present invention provides a ceramide production promoter, which contains one or two selected from the group consisting of purple daisy and lavender as active ingredients.

The Echinacea purpurea and lavender of the present invention have a high effect of promoting ceramide production, in particular, acylceramide production.

Purple varietal has the effect of promoting the expression of the ABCA12 gene involved in the synthesis and secretion of intercellular lipids such as ceramide, the effect of promoting the expression of the SGMS1 gene, the effect of promoting the expression of the ELOVL4 gene involved in the synthesis of acylceramide, the effect of promoting the expression of the CYP4F22 gene, and the effect of promoting the expression of the CERS3 gene. effect, PNPLA1 gene expression promoting effect, and ABHD5 gene expression promoting effect.

Lavender exhibits the effect of promoting the expression of the ABCA12 gene involved in the synthesis and secretion of ceramide, the effect of promoting the expression of the GBA gene, the effect of promoting the expression of the PNPLA1 gene involved in the synthesis of acylceramide, and the effect of promoting the expression of the ABHD5 gene.

EMBODIMENT OF THE INVENTION Below, the form for implementing this invention is demonstrated.

Echinacea purpurea used in the present invention is a perennial plant belonging to the family Asteraceae, and is also called Echinacea, Echinacea, or Echinacea. Although various parts such as leaves, stems, flowers, roots, etc., and the whole plant can be used for the purple varietal, it is preferable to use the roots.

When used, purple varietal can be used in a fresh state or in a dried form by adding it to drinks and the like. Moreover, it can also be used as an extract obtained by extracting Purple Forsythia with a solvent.

Although the extract may be subjected to extraction in its raw state, in consideration of extraction efficiency, it is preferable to carry out extraction after performing treatments such as chopping, drying, and pulverizing. Extraction is performed by immersion in an extraction solvent. To increase extraction efficiency, stirring may be performed or homogenization may be performed in an extraction solvent. It is appropriate that the extraction temperature be from about 5° C. to a temperature below the boiling point of the extraction solvent. Although the extraction time varies depending on the type of extraction solvent and extraction temperature, it is appropriate to set it to about 4 hours to 14 days.

In addition to water, extraction solvents include lower alcohols such as methanol, ethanol, propanol, and isopropanol, polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, dipropylene glycol, and glycerin, and ethers such as ethyl ether and propyl ether. Polar organic solvents such as esters, esters such as ethyl acetate and butyl acetate, and ketones such as acetone and ethyl methyl ketone can be used, and one or more of these may be selected and used. Alternatively, physiological saline, phosphate buffer, phosphate buffered saline, etc. may be used.

The extract of the above-mentioned plant in the above-mentioned solvent can be contained in the composition according to the present invention as it is, but it can be concentrated and dried and then re-dissolved in water or a polar solvent, or it can be used to improve skin physiological function. It may be used after performing purification treatments such as decolorization, deodorization, desalting, etc. or fractionation treatment by column chromatography within a range that does not impair the effect. For preservation, it can also be freeze-dried after purification and dissolved in a solvent before use. It can also be used by being encapsulated in vesicles such as liposomes, microcapsules, and the like.

In addition, in the present invention, it is preferable to use one or more selected from water, ethanol, and 1,3-butylene glycol as the extracting solvent for C. oleracea, and it is more preferable to use an ethanol aqueous solution.

The lavender (L. . For lavender, various parts such as leaves, stems, flowers, flower spikes, roots, and the whole plant can be used, but it is preferable to use flowers or flower spikes.

Lavender can be used in a fresh state or dried by adding it to drinks and the like. Moreover, it can also be used as a lavender extract obtained by extracting lavender using a solvent. The method for obtaining the lavender extract is the same as the method for obtaining the lavender extract. As the lavender extraction solvent, it is preferable to use one or more selected from water, ethanol, and 1,3-butylene glycol.

One or two selected from the group consisting of C. oleracea and lavender have an excellent ceramide production-promoting effect, particularly an acylceramide production-promoting effect, and can be used as a ceramide production promoter or an acylceramide production promoter.

Purple Forsythium has the effect of promoting ABCA12 gene expression, SGMS1 gene expression, ELOVL4 gene expression, CYP4F22 gene expression, CERS3 gene expression, and PNPLA1 gene expression, which are involved in ceramide synthesis/secretion and acylceramide synthesis. , and exhibits ABHD5 gene expression promoting effect, ABCA12 gene expression promoter, SGMS1 gene expression promoter, ELOVL4 gene expression promoter, CYP4F22 gene expression promoter, CERS3 gene expression promoter, PNPLA1 gene expression promoter, and ABHD5 gene. It can be used as an expression promoter.

Lavender exerts an ABCA12 gene expression promoting effect, a GBA gene expression promoting effect, a PNPLA1 gene expression promoting effect, and an ABHD5 gene expression promoting effect, which are involved in ceramide synthesis/secretion and acylceramide synthesis, and is an ABCA12 gene expression promoter. It can be used as a GBA gene expression promoter, a PNPLA1 gene expression promoter, and an ABHD5 gene expression promoter.

The ceramide production promoter, which is characterized by containing one or two selected from the group consisting of lavender and lavender, can be used alone; By blending it into various compositions such as cosmetics, it is possible to obtain a composition that has a ceramide production-promoting effect, particularly an acylceramide production-promoting effect.

The ceramide production promoter, which is characterized by containing one or two selected from the group consisting of lavender and lavender, is not limited in any way as to its form and the presence or absence of other ingredients. Regarding the form, any form such as liquid, paste, gel, solid, powder, etc. can be selected depending on the purpose, etc., and the necessary vehicle (excipient), solvent, Other common additives (antioxidants, colorants, dispersants, etc.) can be optionally included.

A composition that contains a ceramide production promoter characterized by containing one or two selected from the group consisting of lavender and lavender can take any form. When the composition is a skin cosmetic, a hair cosmetic, a cleansing agent, etc., it can be provided as a solubilized system such as a lotion, a dispersion system such as a calamine lotion, or an emulsified system such as a cream or milky lotion. Furthermore, it can be provided in various dosage forms such as an aerosol filled with a propellant, an ointment, and a poultice.

Specifically, various cosmetics such as milky lotions, creams, lotions, lotions, packs, serums, cleaning products, and makeup cosmetics; solutions, ointments, powders, granules, aerosols, patches, poultices, etc. Examples include various forms of cosmetics, quasi-drugs, and external medicines.

In the case of oral pharmaceuticals, they can be in common dosage forms such as liquid preparations such as drinks and drips, solid preparations such as gum and candy, capsules, powders, granules, and tablets.

In addition to the above, compositions containing one or two selected from the group consisting of lavender and lavender may also contain pharmaceuticals, quasi-drugs, skin cosmetics, hair cosmetics, and cleaning products, depending on the intended use and necessity. and any other ingredients normally added to oral pharmaceuticals, such as water, oily ingredients, humectants, powders, pigments, emulsifiers, solubilizers, gelling agents, detergents, ultraviolet absorbers, and anti-inflammatory agents. , a thickener, a pH adjuster, a chelating agent, a drug (medicinal ingredient), a fragrance, a resin, an antibacterial and fungicidal agent, an antioxidant, an alcohol, and the like can be appropriately blended. Furthermore, it is possible to use the composition in combination with other moisturizing agents, anti-aging agents, whitening agents, antioxidants, slimming agents, or various plants/fungi or extracts thereof, as long as the effects of the present invention are not impaired.

EXAMPLES Hereinafter, the present invention will be specifically explained with reference to Examples, but the scope of the present invention is not limited thereby.

[Purple Balensis Extract Powder]
30g of the dried roots of P. aeruginosa were cut into thin pieces and soaked in 600g of a 50% by volume ethanol aqueous solution at room temperature for 48 hours. After filtration, the solvent was distilled off to obtain about 5 g of powdered Purple Forsythium extract.

［Purple Valencia Extract''
0.5 g of powdered Purple Forsythium extract was dissolved in 99.5 g of a 50% by mass 1,3-butylene glycol aqueous solution to obtain a Purple Forsythium extract.

[Lavender extract]
Lavender flowers were dried, cut into pieces, immersed in a 50% by mass 1,3-butylene glycol solution, and filtered to obtain a lavender extract.

[ABCA12 gene expression promoting effect, SGMS1 gene expression promoting effect, ELOVL4 gene expression promoting effect, CYP4F22 gene expression promoting effect, CERS3 expression promoting effect, PNPLA1 gene expression promoting effect, ABHD5 gene expression promoting effect]
Human epidermal keratinocytes were seeded at 3*10 ⁵ cells/well in a 6-well plate, and cultured overnight in Humedia-KG2 medium (manufactured by Kurabo Industries, Ltd.). The medium was replaced with a medium in which 0.5 mg/mL of Purple Forsythiasis extract powder and 0.5 volume % of lavender extract were added and dissolved, and cultured at 37° C. in a 5% CO ₂ incubator for 24 hours. RNA was extracted from the collected cells using a commercially available RNA extraction kit (Quick Gene RNA Cultured Cell HC Kit S), and after cDNA synthesis, gene expression was confirmed by real-time PCR using the cyber green method using the following primers. did. Note that GAPDH was used as an internal standard. Table 2 shows the effects of purple varietal extract powder, and Table 3 shows the effects of lavender extract. The mRNA expression level was expressed as a relative value, with the expression level without the addition of each extract being set to 1. Furthermore, the significant difference from the case without addition was calculated by t-test, and 5% significance is indicated by * and 1% significance is indicated by **.

As shown in Table 2, Purple Forsythiasis extract powder is present on the membrane of the lamellar granules in the epidermal granule cells, and is responsible for the ABCA12 gene, which plays an important role in the secretion of intercellular lipids, and is responsible for the synthesis of ceramide to sphingomyelin. SGMS1 gene, ELOVL4 gene, which is essential for the biosynthesis of fatty acids with carbon numbers of 28 or more in the epidermis, CYP4F22 gene, which hydroxylates the ω-position using very long chain fatty acids as a substrate, specifically reacts with very long chain acyl-CoA, and produces ceramide. ceramide synthesis is promoted by promoting the expression of the CERS3 gene, which synthesizes linoleic acid from triglyceride, the PNPLA1 gene, which biosynthesizes acylceramide by forming an ester bond between linoleic acid derived from ceramide and triglyceride, and the ABHD5 gene, which is involved in the supply of linoleic acid from triglyceride. It was particularly effective in promoting acylceramide biosynthesis.

As shown in Table 3, lavender extract contains the ABCA12 gene, which exists on the membrane of lamellar granules in epidermal granule cells and plays an important role in the secretion of intercellular lipids, the GBA gene involved in glucosylceramidase, and ceramide. Linoleic acid derived from triglyceride forms an ester bond and promotes the expression of the PNPLA1 gene, which biosynthesizes acylceramide, and the ABHD5 gene, which is involved in the supply of linoleic acid from triglyceride, promoting ceramide synthesis, especially acylceramide biosynthesis. It had an excellent promoting effect.

[Ceramide production promotion effect in human 3D cultured skin model]
A human three-dimensional cultured skin model LabCyte EPIMODEL24 (manufactured by J-TEC) was cultured in an assay medium (manufactured by J-TEC) at 37° C. in a 5% CO ₂ incubator for 24 hours.
After removing the medium from each well, an assay medium containing 0.5 mg/mL of P. aeruginosa extract powder was added to the outside of the 0.5 mL culture cup, and cultured in a CO ₂ incubator for 24 hours.
After removing the medium from each well, the cells were cultured for 48 hours in an assay medium (manufactured by J-TEC) containing no sample.
The skin model was washed three times with PBS, and lipids were extracted by the Bligh-Dyer method. Note that the obtained solid substance was dissolved in 50 μL of a chloroform:methanol (volume ratio 2:1) solution.
Quantification of lipid samples was performed using HPTLC. The test method using HPTLC is shown below.
A filter paper was placed on the wall of the TLC developing tank, and a developing solution (chloroform:methanol:acetic acid (volume ratio 190:9:1) solution) was added to the extent that it did not touch the spot positions of the HPTLC plate.
Using a microsyringe, 5 μL of each lipid sample was spotted onto a 1.5 cm portion from the bottom of the glass of the HPTLC plate.
In addition, as standard products, Ceramide I (manufactured by Matreya), Ceramide II (manufactured by Avanti Polar Lipids), and Ceramide III (manufactured by SIGMA) were prepared at concentrations of 15.625, 31.25, 62.5, 125, and 250 μg/mL. They were dissolved in a chloroform:methanol (volume ratio 2:1) solution, and 5 μL of each was spotted.
The HPTLCC plate was gently placed in a developing tank and developed to the top (8 cm from the spot). After drying the developing solvent, a 10% by mass CuSo ₄ -8% by volume H ₃ PO ₄ solution was evenly sprayed onto the HPTLC plate and heated at 150° C. for 10 minutes.
After the HPTLC plate was cooled to room temperature, a photograph of the plate was taken using a digital camera (EOS5D Mark II manufactured by Canon) installed in a cabinet for TLC photography.
Using JustTLC (manufactured by SWEDAY) from the obtained image data, a calibration curve regarding the signal intensity of each band was created, and the amount of ceramide in each band was calculated.
The amount of ceramide is shown in Table 4 as a relative value, with the amount of ceramide in the case without addition of Purple Forsythia extract being 1.

As shown in Table 4, the powdered Purple Forsythium extract exhibited a higher ceramide production promoting effect than when the same extract was not added.

Claims (1)

A PNPLA1 gene expression promoter whose active ingredient is Purple Forsythium.