Agent for enhancing expression of pleiotrophin receptor

Abstract

To provide a novel composition capable of promoting expression of a pleiotrophin receptor.SOLUTION: An agent for enhancing pleiotrophin receptor expression comprising, as an active ingredient, one or two or more plant extracts of plants selected from the group consisting of Ruscus aculeatus, Olea europaea, and Plantago major.SELECTED DRAWING: Figure 2

Description

[Technical Field]

[0001] The present invention relates to a pleiotrophin receptor expression enhancer, an agent for inducing extracellular matrix degradation and / or neoplasia, a skin fiber structure improving agent, an external skin composition for enhancing pleiotrophin receptor expression, an external skin composition for improving skin condition, and a method for screening a skin condition improving agent. [Background technology]

[0002] Good skin condition not only increases self-satisfaction and self-confidence and has a positive impact on daily life, but also affects one's impression and trustworthiness, so there is a demand for products that help keep skin in good condition.

[0003] Skin viscoelasticity / elasticity is used as one of the indicators of skin condition (Non-Patent Document 1). Therefore, topical preparations that improve skin viscoelasticity / elasticity have been developed (Patent Document 1).

[0004] It is also known that the dermis influences the viscoelasticity / elasticity of the skin (Non-Patent Document 2). The dermis deteriorates with age, and various mechanisms for regenerating the dermis have been proposed (Patent Document 2). However, no satisfactory solution has yet been provided.

[0005] Here, one example of a natural phenomenon in which new dermis is created in the living body is when a hair follicle regresses into the catagen phase and then regenerates in the dermal area where the hair follicle once existed (Figure 1). The catagen phase is one of three stages in the "hair cycle," which is a cycle of repeated hair growth and shedding, and is divided into the anagen phase, catagen phase, and telogen phase (Non-Patent Document 3).

[0006] Recently, it has been reported that changes in gene expression in various cells around hair follicles are linked to the regeneration of the dermis around the hair follicle during hair follicle regression (Non-Patent Document 4). [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Japanese Unexamined Patent Application Publication No. 2021-116276 [Non-Patent Document]

[0008] [[ID=I2]] [Non-Patent Document 1] Correlation between a Cutometer and quantitative evaluation using Moire topography in age-related skin elasticity[online], February 2007, [Search Date: April 24, 2023], <URL:https: / / doi.org / 10.1111 / j.1600-0846.2007.00224.x> [Non-Patent Document 2] Molecular Mechanisms of Dermal Aging and Antiaging Approaches[online], May 2019, [Search Date: April 24, 2023], <URL:https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC6540032 / > [Non-Patent Document 3] New Dermatology, 3rd Edition (Nakayama Shoten)[online], February 2018, [Search Date: April 24, 2023], <URL:https: / / www.derm-hokudai.jp / wp / wp-content / uploads / 2021 / 12 / 1-08.pdf> [Non-Patent Document 4] Constructing a Single-Cell Transcriptome Atlas of the Hair Cycle to Decipher Tissue Regeneration Dynamics in Human Skin[online], January 2024, [Search Date: April 24, 2023], <URL:http: / / dx.doi.org / 10. / ssrn.<4690956> [Summary of the Invention] [Problem to be solved by the invention]

[0009] The present inventors independently studied the effects of various ligands described in Non-Patent Document 4 on dermal regeneration and found that pleiotrophin (PTN) is particularly effective in dermal regeneration (Figure 2). In other words, it was found that dermal regeneration can be induced by enhancing the expression of pleiotrophin or pleiotrophin receptor.

[0010] An objective of the present invention is to provide a novel composition that enhances the expression of pleiotrophin receptors. [Means for solving the problem]

[0011] The present invention, which solves the above-mentioned problems, provides a pleiotrophin receptor expression enhancer, which contains as an active ingredient one or more plant extracts selected from the group consisting of Ruscus aculeatus, olive (Olea europaea), and plantain (Plantago major). According to the above invention, the expression of the pleiotrophin receptor can be enhanced, and by enhancing the expression of the pleiotrophin receptor, regeneration of the dermis can be induced.

[0012] In a preferred embodiment of the present invention, the pleiotrophin receptor is syndecan-4.

[0013] In a preferred embodiment of the present invention, the present invention is a pleiotrophin receptor expression enhancer for improving skin conditions.

[0014] In addition, a preferred embodiment of the present invention is a pleiotrophin receptor expression enhancer that improves one or more selected from skin wrinkles, sagging, and skin viscoelasticity and / or elasticity.

[0015] The present invention also relates to an agent for inducing the degradation and / or regeneration of extracellular matrix, which contains as an active ingredient one or more plant extracts selected from the group consisting of Ruscus aculeatus, olive (Olea europaea), and plantain (Plantago major).

[0016] The present invention also relates to a skin fiber structure improving agent containing, as an active ingredient, one or more plant extracts selected from the group consisting of Ruscus aculeatus, olive (Olea europaea), and plantain (Plantago major).

[0017] The present invention also relates to an external skin composition for enhancing pleiotrophin receptor expression, which contains, as an active ingredient, one or more plant extracts selected from the group consisting of Ruscus aculeatus, Olea europaea, and Plantago major.

[0018] The present invention also relates to an external skin composition for improving skin conditions, which comprises a pleiotrophin receptor expression enhancer and a pleiotrophin expression enhancer, and the active ingredient is one or more plant extracts selected from Ruscus aculeatus, Olea europaea, and Plantago major.

[0019] The present invention also provides a method for screening for a skin condition improving agent, using the effect of enhancing pleiotrophin receptor expression as an indicator.

[0020] In a preferred embodiment of the present invention, the screening method is such that the test substance is judged to be a more excellent skin condition-improving agent as the test substance exhibits a higher effect of enhancing the expression of pleiotrophin receptors cultured in the presence of the test substance.

[0021] In a preferred embodiment of the present invention, the pleiotrophin receptor is syndecan-4. [Effects of the Invention]

[0022] According to the present invention, it is possible to enhance the expression of pleiotrophin receptors, induce the degradation and / or regeneration of extracellular matrix, or improve the structure of skin fibers. Furthermore, according to the present invention, screening of skin condition improving agents can be carried out. [Brief explanation of the drawings]

[0023] [Figure 1] Diagram of the hair cycle and dermal regeneration after hair follicle regression [Figure 2] Conceptual diagram of pleiotrophin and dermal regeneration [Figure 3] Graph showing the results of analysis of the functions of genes whose expression is increased by pleiotrophin [Figure 4] Graph showing verification results of collagen gene expression levels [Figure 5] Image showing the results of observing collagen structure [Figure 6] Graph showing the results of verifying the gene expression of syndecan-4 by plant extracts [Figure 7] Graph showing the results of verifying the expression of syndecan-4 genes by the complex extract [Figure 8] Graph showing the results of verifying the effect of brown algae extract 2 on pleiotrophin gene expression [Figure 9] Graph showing the results of verifying pleiotrophin gene expression by the composite extract [Figure 10] Graph showing the results of verifying pleiotrophin gene expression by plant extracts DETAILED DESCRIPTION OF THE INVENTION

[0024] Preferred embodiments of the present invention will be described below, but it goes without saying that the technical scope of the present invention is not limited to the following embodiments.

[0025] <1> Agent The present invention relates to a pleiotrophin receptor expression enhancer, which comprises, as an active ingredient, one or more plant extracts selected from the group consisting of Ruscus aculeatus, Olive (Olea europaea), and Plantain (Plantago major). According to the above invention, the expression of the pleiotrophin receptor can be enhanced, and by enhancing the expression of the pleiotrophin receptor, regeneration of the dermis can be induced.

[0026] <1-1> Active ingredient The plant extracts used in the present invention can be extracted using the whole plant or other parts such as the plant itself, above-ground parts, rhizomes, trunks, leaves, stems, spikes, flower buds, and seeds. However, it is preferable to crush or shred these materials beforehand to improve extraction efficiency. Suitable examples of extraction solvents include one or more polar solvents selected from water, alcohols such as ethanol, isopropyl alcohol, and butanol, polyhydric alcohols such as 1,3-butylene glycol and propylene glycol, ketones such as acetone and methyl ethyl ketone, and ethers such as diethyl ether and tetrahydrofuran. Water, alcohols, or a mixture of water and alcohols is preferably used as the extraction solvent.

[0027] Specific extraction methods include, for example, adding 1 to 30 parts by mass of solvent to 1 mass of the part of the plant body or its dried material used for extraction, immersing for several days at room temperature or for several hours at a temperature near the boiling point, cooling to room temperature, removing insoluble matter and / or solvent as desired, and fractionating and purifying by column chromatography or the like, but the extraction method is not limited to this.

[0028] The plant extract of Ruscus aculeatus is preferably Ruscus aculeatus root extract. As the extraction solvent, water or an ethanol solution can be preferably used.

[0029] The plant extract of olive (Olea europaea) is preferably olive leaf extract. As the extraction solvent, water or an ethanol solution can be preferably used.

[0030] The plant extract of Plantago major is preferably Plantago major seed extract. As the extraction solvent, water or an ethanol solution can be preferably used.

[0031] <1-2>Applications A preferred embodiment of the present invention is a pleiotrophin receptor expression enhancer comprising, as an active ingredient, one or more plant extracts selected from the group consisting of Ruscus aculeatus, Olea europaea, and Plantago major. The pleiotrophin receptor is preferably a proteoglycan, more preferably a syndecan family protein, and even more preferably syndecan 4 (SDC4).

[0032] By increasing the expression of pleiotrophin receptors, fibroblasts become more susceptible to pleiotrophin, which leads to the synthesis and / or degradation of the extracellular matrix and structural improvement in the fibroblasts, thereby improving skin condition.

[0033] Specifically, the active ingredient can improve one or more of the following: wrinkles, sagging skin, and skin viscoelasticity and / or elasticity.

[0034] A preferred embodiment of the present invention is an agent for inducing degradation and / or neoplasia of extracellular matrix, which contains as an active ingredient one or more plant extracts selected from the group consisting of Ruscus aculeatus, Olive (Olea europaea), and Plantago major.

[0035] In a preferred embodiment of the present invention, the agent is a collagen degradation and / or neogenesis inducer, which contains as an active ingredient one or more plant extracts selected from the group consisting of Ruscus aculeatus, Olive (Olea europaea), and Plantain (Plantago major).

[0036] The above-mentioned agents for inducing degradation and / or neoplasia of extracellular matrix can enhance expression of pleiotrophin receptors. Enhanced expression of pleiotrophin receptors makes fibroblasts more susceptible to pleiotrophin, leading to the synthesis and / or degradation of extracellular matrix in fibroblasts.

[0037] A preferred embodiment of the present invention is a skin fiber structure improving agent containing, as an active ingredient, one or more plant extracts selected from the group consisting of Ruscus aculeatus, Olive (Olea europaea), and Plantago major.

[0038] A preferred embodiment of the present invention is a dermal fiber structure improving agent containing, as an active ingredient, one or more plant extracts selected from the group consisting of Ruscus aculeatus, Olive (Olea europaea), and Plantago major.

[0039] In a preferred embodiment of the present invention, the collagen improving agent contains, as an active ingredient, one or more plant extracts selected from the group consisting of Ruscus aculeatus, Olive (Olea europaea), and Plantago major.

[0040] Here, the fibrous structure of the skin is composed of fibers such as collagen, as shown in Figure 5. In this specification, improving the fibrous structure of the skin means making this fibrous structure oriented to some extent.

[0041] The skin fiber structure improving agent can enhance the expression of pleiotrophin receptors. Enhanced expression of pleiotrophin receptors makes fibroblasts more susceptible to pleiotrophin. Enhanced pleiotrophin receptivity by fibroblasts improves skin fiber structure.

[0042] A preferred embodiment of the present invention is a composition for external application to the skin for enhancing pleiotrophin receptor expression, which contains as an active ingredient one or more plant extracts selected from the group consisting of Ruscus aculeatus, olive (Olea europaea), and plantain (Plantago major).

[0043] <1-3>Usage / Capacity In the present invention, the undiluted extract obtained by extraction with any solvent may be used as is as the agent of the present invention, or the undiluted extract may be diluted to any concentration and then used as the agent of the present invention. The agent according to the present invention is preferably applied to the dermis.

[0044] The total content of one or more extracts selected from the plant extracts of the present invention in the agent of the present invention is typically 0.00001% by mass or more, based on the dry mass, preferably 0.0001% by mass or more, and more preferably 0.001% by mass or more.

[0045] Furthermore, the total content of one or more extracts selected from the plant extracts of the present invention in the agent of the present invention is typically 80% by mass or less, based on the dry mass, preferably 50% by mass or less, more preferably 30% by mass or less, even more preferably 10% by mass or less, and even more preferably 1% by mass or less.

[0046] Furthermore, the total content of one or more active ingredients selected from the plant extracts of the present invention depends on the dosage form, and the amount applied to the skin per application (application amount) is usually 2000 mg or less, preferably 1000 mg or less, and more preferably 500 mg or less, as the dry mass of the extract.

[0047] <2> Composition for topical application to the skin for enhancing pleiotrophin receptor expression A preferred embodiment of the present invention is a composition for external application to the skin for enhancing pleiotrophin receptor expression, which contains, as an active ingredient, one or more plant extracts selected from the group consisting of Ruscus aculeatus, Olive (Olea europaea), and Plantain (Plantago major).

[0048] The composition for external application to skin for enhancing pleiotrophin receptor expression of the present invention is preferably formulated into an external or oral preparation in appropriate combination with any ingredients used in formulation. Examples of external preparations include cosmetics, quasi-drugs, external skin medicines, etc. There are no particular limitations on the dosage form.

[0049] Furthermore, when the present invention is in the form of a composition for external use on the skin, it is also preferable that the present invention be in a form labeled with an indication of use, such as "for enhancing pleiotrophin receptor expression," "for improving skin wrinkles," "for improving skin sagging," "for improving skin viscoelasticity," "for improving skin elasticity," "for inducing degradation and / or regeneration of extracellular matrix," or "for improving skin fiber structure."

[0050] The "indication" includes all indications that have the function of informing consumers of the intended use. In other words, any indication that can recall or infer the intended use falls under the category of "indication," regardless of the purpose of the indication, the content of the indication, the object or medium on which it is displayed, etc. Furthermore, the phrase "labeled" means that there is a labeling action that aims to associate the label with the topical skin composition (product) etc.

[0051] The act of labeling is preferably one that allows consumers to directly recognize the intended use. Specific examples include the act of describing the intended use on the product or its packaging according to the present invention, and the act of describing the intended use in product advertisements, price lists, or transaction documents (including those provided by electromagnetic means).

[0052] When the external skin composition for inhibiting gene expression of the present invention is used as a cosmetic, in addition to the above-mentioned components, it may contain whitening components, components for improving wrinkles and / or sagging, anti-inflammatory components, etc.

[0053] There are no particular limitations on the whitening ingredient, as long as it is one that is commonly used in cosmetics. Examples of water-soluble whitening ingredients include 4-n-butylresorcinol, ascorbic acid glucoside, 3-O-ethyl ascorbic acid, tranexamic acid, ellagic acid, arbutin, nicotinamide, and pantothenyl alcohol. Examples of oil-soluble whitening ingredients include 1-triphenylmethylpiperidine, 1-triphenylmethylpyrrolidine, 2-(triphenylmethyloxy)ethanol, 2-(triphenylmethylamino)ethanol, 2-(triphenylmethyloxy)ethylamine, triphenylmethylamine, triphenylmethanol, triphenylmethane, aminodiphenylmethane, N-(o-toluoyl)cysteic acid, N-(m-toluoyl)cysteic acid, N-(p-toluoyl)cysteic acid, and N-(p-methoxybenzoyl)cysteic acid. Further preferred examples of other whitening ingredients include N-benzoylserine, N-(p-methylbenzoyl)serine, N-(p-ethylbenzoyl)serine, N-(p-methoxybenzoyl)serine, N-(p-fluorobenzoyl)serine, N-(p-trifluoromethylbenzoyl)serine, N-(2-naphthoyl)serine, N-(4-phenylbenzoyl)serine, N-(p-methylbenzoyl)serine methyl ester, N-(p-methylbenzoyl)serine ethyl ester, N-(2-naphthoyl)serine methyl ester, N-benzoyl-O-methylserine, N-(p-methylbenzoyl)-O-methylserine, N-(p-methylbenzoyl)-O-acetylserine, and N-(2-naphthoyl)-O-methylserine.

[0054] The other anti-wrinkle ingredients are not particularly limited as long as they are commonly used in cosmetics. Examples include vitamin A or its derivatives such as retinol, retinal, retinoic acid, tretinoin, isotretinoin, tocopherol retinoate, retinol palmitate, and retinol acetate. Other examples include ursolic acid benzyl ester, ursolic acid phosphate ester, betulinic acid benzyl ester, benzilic acid phosphate ester, and nicotinamide. The content of other anti-wrinkle ingredients in cosmetics besides the neutrophil chemotactic factor gene expression inhibitor is typically 0.0001 to 30% by mass, preferably 0.001 to 10% by mass, and more preferably 0.01 to 5% by mass (dry mass in the case of an extract).

[0055] Examples of anti-inflammatory components include kurarinone, glabridin, glycyrrhizinic acid, dipotassium glycyrrhizinate, glycyrrhetinic acid, alkyl glycyrrhetinate, pantothenyl alcohol, etc. Among these, preferred examples of oil-soluble components include glabridin, glycyrrhetinic acid and salts thereof, and alkyl glycyrrhetinate and salts thereof. The content of the anti-inflammatory component in the cosmetic is usually 0.01 to 30% by mass, preferably 0.1 to 10% by mass, and more preferably 1 to 5% by mass (dry mass in the case of an extract).

[0056] It is also preferable to use extracts derived from animals and plants that are generally used in medicines, cosmetics, foods, etc.For example, Akebia extract, Asunaro extract, Asparagus extract, Avocado extract, Hydrangea extract, Almond extract, Arnica extract, Aronia extract, Apricot extract, Ginkgo extract, Fennel extract, Aralia extract, Aralia udo extract, Eleuthero extract, Enmeiso extract, Phellodendron bark extract, Panax ginseng extract, Nettle extract, Pueraria lobata extract, Pueraria lobata extract, Chamomilla recutita extract, Carrot extract, Artemisia capillaris extract, Licorice extract, Kiwi extract, Cucumber extract, Guava extract, Gardenia extract, Sasa veitchii extract, Kuru extract Rice extract, black rice extract, chlorella extract, mulberry extract, cinnamon root extract, Alpinia zerumbet extract, gentian extract, geranium extract, rice extract, fermented rice extract, fermented rice bran extract, rice germ oil, salvia extract, soapwort extract, bamboo extract, sanshou extract, Japanese pepper extract, shiitake mushroom extract, rehmannia root extract, lithospermum root extract, perilla extract, linden extract, meadowsweet extract, ginger extract, calamus root extract, horsetail extract, stevia extract, fermented stevia, elderberry extract, yaw wood Lysom extract, peppermint extract, sage extract, mallow extract, cnidium extract, swertia bristle extract, mulberry extract, rhubarb extract, soybean extract, tallow extract, thyme extract, dandelion extract, clove extract, chili pepper extract, angelica extract, calendula extract, peach kernel extract, tomato extract, natto extract, carrot extract, garlic extract, hibiscus extract, burdock extract, lotus extract, parsley extract, birch extract, witch hazel extract, jasmine extract, cypress extract, loquat extract, Preferred examples of extracts include dandelion extract, butterbur extract, poria extract, loofah extract, peppermint extract, linden extract, pine extract, skunk cabbage extract, melissa extract, mozuku extract, cornflower extract, eucalyptus extract, lily extract, coix seed extract, mugwort extract, lavender extract, apple extract, rooibos tea extract, lychee extract, lettuce extract, forsythia extract, astragalus extract, rosemary extract, Roman chamomile extract, royal jelly extract, and burnet extract.

[0057] The content (dry mass) of the optional animal and plant-derived extract in the cosmetic is usually 0.01 to 30 mass %, preferably 0.1 to 10 mass %, more preferably 0.3 to 3 mass %.

[0058] In addition to the above-mentioned components, optional components commonly used in cosmetics include polyethylene glycol, glycerin, 1,3-butylene glycol, erythritol, sorbitol, xylitol, maltitol, propylene glycol, dipropylene glycol, diglycerin, isoprene glycol, polyols such as 1,2-pentanediol, 2,4-hexylene glycol, 1,2-hexanediol, 1,2-octanediol, and cyclohexylglycerin, fatty acid soaps ( Anionic surfactants such as sodium laurate, sodium palmitate, potassium lauryl sulfate, and alkyl sulfate triethanolamine ether; cationic surfactants such as stearyltrimethylammonium chloride, benzalkonium chloride, and laurylamine oxide; imidazoline-based amphoteric surfactants (2-cocoyl-2-imidazolinium hydroxide-1-carboxyethyloxy disodium salt, etc.); betaine-based surfactants (alkyl betaine, amido betaine, sulfobetaine, etc.); amphoteric surfactants such as acyl methyl taurine; sorbitan fatty acid esters (sorbitan monostearate, sorbitan sesquioleate, etc.); glycerin fatty acids (glycerin monostearate, etc.); propylene glycol fatty acid esters (propylene glycol monostearate, etc.); hydrogenated castor oil derivatives; glycerin alkyl ether; POE sorbitan Tan fatty acid esters (POE sorbitan monooleate, polyoxyethylene sorbitan monostearate, etc.), POE sorbitan fatty acid esters (POE-sorbitan monolaurate, etc.), POE glycerin fatty acid esters (POE-glycerin monoisostearate, etc.), POE fatty acid esters (polyethylene glycol monooleate, POE distearate, etc.), POE alkyl ethers (POE 2-octyldodecyl ether, etc.), POE alkyl phenyl ethers (POE nonylphenyl ether, etc.), Pluronic types, POE·POP alkyl ethers (POE·POP 2-decyltetradecyl ether, etc.), Tetronics, POE castor oil·hydrogenated castor oil derivatives (POE castor oil, POE hydrogenated castor oil, etc.), sucrose fatty acid esters, nonionic surfactants such as alkyl glucosides, sodium pyrrolidone carboxylate, lactic acid,Moisturizing ingredients such as sodium lactate; powders such as mica, talc, kaolin, synthetic mica, calcium carbonate, magnesium carbonate, anhydrous silicic acid (silica), aluminum oxide, and barium sulfate, which may be surface-treated; inorganic pigments such as cobalt oxide, ultramarine, Prussian blue, and zinc oxide, which may be surface-treated; composite pigments such as sintered iron oxide and titanium dioxide, which may be surface-treated; pearlizing agents such as titanium mica, fish phosphate foil, and bismuth oxychloride, which may be surface-treated; Red No. 202, Red No. 228, Red No. 226, Yellow No. 4, Blue No. 404, Yellow No. 5, Red No. 505, Red No. 230, Red No. 223, Orange No. 201, Red No. 213, Yellow No. 204, Yellow No. 203, and Blue No. 505, which may be laked. Examples of suitable organic pigments include organic pigments such as No. 1, Green No. 201, Purple No. 201, and Red No. 204; organic powders such as polyethylene powder, polymethyl methacrylate, nylon powder, and organopolysiloxane elastomer; lower alcohols such as ethanol and isopropanol; vitamin A or a derivative thereof, vitamin B6 hydrochloride, vitamin B6 tripalmitate, vitamin B6 dioctanoate, vitamin B2 or a derivative thereof, vitamin B12, and vitamin B15 or a derivative thereof; vitamin E such as α-tocopherol, β-tocopherol, γ-tocopherol, and vitamin E acetate; vitamin D, vitamin H, pantothenic acid, pantethine, and pyrroloquinoline quinone.

[0059] <3> External skin composition for improving skin conditions A preferred embodiment of the present invention is a composition for external application to the skin for improving skin conditions, which comprises a pleiotrophin receptor expression enhancer and a pleiotrophin expression enhancer, and the active ingredient is one or more plant extracts selected from the group consisting of Ruscus aculeatus, olive (Olea europaea), and plantain (Plantago major). By including both a pleiotrophin receptor expression enhancer and a pleiotrophin expression enhancer, the effect of pleiotrophin can be further improved. Specifically, it is possible to obtain higher effects of improving skin condition, inducing degradation and / or regeneration of extracellular matrix, and improving skin fiber structure.

[0060] The pleiotrophin receptor expression enhancer is as described above.

[0061] In the topical skin composition for improving skin condition of the present invention, it is preferable to use a brown algae extract as an active ingredient as a pleiotrophin expression enhancer. The brown algae extract may be an extract of one or more brown algae selected from Fucus serratus, Sargassum fuluvellum, Cystoseira tamariscifolia, Ascophyllum nodosum, Hizikia fusiforme, and Sargassum horneri.

[0062] The extract used in the present invention can be extracted using the whole plant or parts such as the plant, above-ground parts, rhizomes, trunks, leaves, stems, spikes, flower buds, and seeds, although it is preferable to crush or shred these beforehand to improve extraction efficiency. Suitable examples of the extraction solvent include one or more polar solvents selected from water, alcohols such as ethanol, isopropyl alcohol, and butanol, polyhydric alcohols such as 1,3-butylene glycol and propylene glycol, ketones such as acetone and methyl ethyl ketone, and ethers such as diethyl ether and tetrahydrofuran. The extraction solvent is preferably water, an alcohol, or a mixture of water and an alcohol.

[0063] Specific extraction methods include, for example, a method in which 1 to 30 parts by mass of solvent is added to 1 mass of the part to be extracted or its dried product, followed by immersion for several days at room temperature or for several hours at a temperature near the boiling point, and then cooling to room temperature, followed by removal of insoluble matter and / or solvent as desired, and fractionation and purification by column chromatography or the like, but the extraction method is not limited to this.

[0064] As a specific extraction method, lixiviation can be used, in which freeze-dried brown algae are exposed to refluxing water, allowing minerals and active ingredients to diffuse into the extraction solvent.

[0065] One specific extraction method that can be used is Vacuum Microwave Hydro Distillation (VMHD), which uses heat and reduced pressure generated by microwaves to destroy brown algae cells, and then collects and concentrates the volatile components within the cells that rise in the vapor stream.

[0066] As a specific extraction method, supercritical carbon dioxide extraction can also be used. In supercritical carbon dioxide extraction, carbon dioxide is subjected to pressure and temperature to turn it into a supercritical gas, and components are extracted from brown algae using the carbon dioxide in the supercritical gas state.

[0067] The extract of Fucus serratus is preferably an extract of the seaweed part of Fucus serratus.

[0068] The extract of Sargassum fuluvellum is preferably an extract of the seaweed part of Sargassum fuluvellum.

[0069] The extract of Cystoseira tamariscifolia is preferably an extract of seaweed parts.

[0070] The extract of Ascophyllum nodosum is preferably an extract of seaweed parts.

[0071] The extract of Hizikia fusiforme is preferably an extract of the seaweed part of Hizikia.

[0072] The extract of Akamoku (Sargassum horneri) is preferably an extract of Akamoku seaweed parts.

[0073] <4> Screening method for skin condition improving agents The present invention also relates to a method for screening for a skin condition improving agent using the effect of enhancing the expression of pleiotrophin receptor as an index.

[0074] As described above, skin conditions can be improved by increasing the expression of the pleiotrophin receptor. Therefore, skin condition-improving agents can be screened using the effect of increasing the expression of the pleiotrophin receptor as an indicator.

[0075] In the present invention, the pleiotrophin receptor is preferably syndecan-4.

[0076] In the screening method of the present invention, the skin condition improving agent to be screened is not particularly limited, and examples of test substances include commercially available compounds (including peptides), known compounds (including peptides), groups of compounds obtained by combinatorial chemistry techniques, extracts derived from living organisms, and mixtures thereof.

[0077] The term "biological extract" refers to not only the extract itself derived from animals or plants, but also a collective term for fractions of extracts, purified fractions, and solvent-removed extracts, fractions, and purified products. Examples of plant extracts include extracts made from wild or cultivated plants, extracts sold as herbal medicine ingredients, and commercially available extracts.

[0078] In the present invention, preferably, the test substance is judged to be a more excellent skin condition-improving agent as the test substance exhibits a higher effect of enhancing the expression of pleiotrophin receptors in cells cultured in the presence of the test substance.

[0079] The present invention preferably comprises adding a test substance to a fibroblast culture system and measuring the effect of enhancing the expression of pleiotrophin receptors in the cells.

[0080] Specifically, the screening method of the present invention makes it possible to determine that a test substance is a superior skin condition improving agent when the test substance has a higher effect of enhancing pleiotrophin receptor expression in fibroblasts cultured in the presence of the test substance.

[0081] Furthermore, preferably, when the effect of enhancing pleiotrophin receptor expression in fibroblasts cultured with the addition of the test substance is statistically significantly greater than the effect of enhancing pleiotrophin receptor expression in fibroblasts cultured without the addition of the test substance, the test substance can be determined to be a candidate for a skin condition improving agent. More specifically, if the effect of enhancing pleiotrophin receptor expression in fibroblasts cultured with the addition of the test substance is more than 1-fold greater than the effect of enhancing pleiotrophin receptor expression in fibroblasts cultured without the addition of the test substance, the test substance can be selected as a candidate for a skin condition improving agent.

[0082] The present invention can also be implemented in a form comprising the steps of culturing fibroblasts in the presence of a test substance, measuring the effect of enhancing pleiotrophin receptor expression in the fibroblasts, and determining that the test substance is a skin condition improving agent if the effect of enhancing pleiotrophin receptor expression is greater than the effect of enhancing pleiotrophin receptor expression in fibroblasts cultured in the absence of the test substance.

[0083] Each step in the embodiment of the present invention will be described in detail below.

[0084] (1)Culture process The culturing step is a step of culturing fibroblasts in the presence of a test substance. The cell culture method is not particularly limited, and adherent culture or suspension culture can be appropriately selected depending on the properties of the cells used.

[0085] Preferably, the fibroblasts are fibroblasts present in the skin.

[0086] The duration of the culture step is not particularly limited, but is preferably 3 hours or more, more preferably 6 hours or more, even more preferably 12 hours or more, and particularly preferably 1 day or more. The upper limit of the culture period is also not particularly limited, but as a guideline, it is preferably 1 month or less, more preferably 3 weeks or less, even more preferably 2 weeks or less, and particularly preferably 1 week or less.

[0087] In the culturing step, culturing is carried out in the presence of a test substance. Specific examples include an embodiment in which the test substance is added to a medium and then cultured. The type of test substance is not limited, and any low molecular weight compound, high molecular weight compound, protein, peptide, nucleic acid, or the like can be applied. The test substance may be a pure compound or a mixture containing two or more components, such as a plant or animal extract.

[0088] In the culturing step, one or more test substances may be used, and when evaluating a synergistic effect due to a combination of test substances, it is preferable to add two or more test substances.

[0089] In the culturing step, cells may be cultured in the absence of the test substance as a control in parallel with the culture in the presence of the test substance. In this case, it is preferable to keep the culture conditions the same except for the presence or absence of the test substance.

[0090] (2) Measurement process The measuring step is a step of measuring the enhancing effect of the pleiotrophin receptor expression in fibroblasts that have been subjected to cell culture. The method for measuring the effect of enhancing the expression of pleiotrophin receptors in fibroblasts is not particularly limited.

[0091] (3) Judgment process The determination step is a step of determining whether or not the test substance is a skin condition improving agent based on the pleiotrophin receptor expression enhancing effect measured in the measurement step. Specifically, the test substance is determined to be a skin condition improving agent when the pleiotrophin receptor expression enhancing effect in fibroblasts cultured in the presence of the test substance is higher than the pleiotrophin receptor expression enhancing effect in fibroblasts cultured in the absence of the test substance.

[0092] Here, the test substance determined to be a skin condition improving agent in the determination step is a pleiotrophin receptor expression enhancer, an extracellular matrix degradation and / or neoplasia inducer, or a skin fiber enhancer. It can be used as a structure improving agent.

[0093] Here, the test substance determined in the determination step may be determined as an active ingredient as is, or may be determined as a candidate active ingredient to be subjected to secondary screening.

[0094] The skin condition improving agent obtained by the screening method of the present invention can be used to improve one or more selected from wrinkles, sagging skin, and skin viscoelasticity and / or elasticity. [Example]

[0095] Example 1 The functions of each gene group whose expression was enhanced by pleiotrophin were analyzed by the following procedure. 1) Human skin fibroblasts were seeded and cultured for 24 hours. 2) Pleiotrophin (final concentration: 300 ng / mL) was added, and the cells were cultured for 6 hours. 3) Total RNA was extracted. 4) Gene expression information was obtained using RNA sequencing. 5) Genes with differential expression were extracted (|rate of change|≧2, p<0.05). 6) The functions of each gene group were analyzed using enrichment analysis.

[0096] The results are shown in Figure 3. As shown in Figure 3, the addition of pleiotrophin significantly increased the expression of genes involved in the synthesis / degradation of extracellular matrix in fibroblasts.

[0097] Since pleiotrophin induces the synthesis / degradation of extracellular matrix in fibroblasts, it has been shown that the expression of pleiotrophin receptors can be used as an indicator to screen for substances that can induce the degradation and / or synthesis of extracellular matrix.

[0098] <Example 2> Changes in collagen gene expression due to pleiotrophin were examined using the following procedure. 1) Human skin fibroblasts were seeded and cultured for 24 hours. 2) Pleiotrophin (final concentration: 300 ng / mL) was added, and the cells were cultured for 48 hours. 3) Total RNA was extracted. 4) The gene expression level of COL1A1 was verified using qPCR.

[0099] The results are shown in Figure 4. As shown in Figure 4, the addition of pleiotrophin significantly increased the expression level of the COL1A1 gene.

[0100] Since the action of pleiotrophin increases the expression level of collagen genes, it was shown that substances that increase the expression level of collagen genes can be screened using the expression of pleiotrophin receptors as an indicator.

[0101] Example 3 The dermal collagen structure was observed using the following procedure. 1) Human facial skin was removed. 2) The dermis with epidermis was immediately cultured at the air-liquid interface in a medium containing pleiotrophin (final concentration 300 ng / mL). 3) After one week of culture, the dermal collagen structure was observed using a scanning electron microscope.

[0102] The results are shown in Figure 5. As shown in Figure 5, the collagen structure is more organized when pleiotrophin is added.

[0103] The improvement of skin fiber structure due to the action of pleiotrophin indicates that substances that improve skin fiber structure can be screened using the expression of pleiotrophin receptors as an indicator.

[0104] Example 4 The gene expression of syndecan-4 by the plant extract was verified by the following procedure. 1) Human skin fibroblasts were seeded and cultured for 24 hours. 2) Plant extracts (final concentration of each: 0.1%) were added and the mixture was cultured for 24 hours. 3) Total RNA was extracted. 4) Syndecan 4 (SDC4) gene expression was verified using qPCR.

[0105] The plant extracts used were Butcher's Broom Root Extract (Butcher's Broom Extract / Maruzen Pharmaceutical Co., Ltd.), Olive Leaf Extract (Olive Leaf Extract BG / Maruzen Pharmaceutical Co., Ltd.), and Plantago Major Seed Extract (Absolage / Ichimaru Pharcos Co., Ltd.).

[0106] The results are shown in Figure 6. As shown in FIG. 6, enhanced gene expression of syndecan-4 was observed in all of the extracts of the root of the kaede, the olive leaf extract, and the seed extract of the plantago major.

[0107] <Example 5> The gene expression of syndecan-4 by the composite extract was verified by the following procedure. 1) Human skin fibroblasts were seeded and cultured for 24 hours. 2) The composite extract was added and cultured for 24 hours. 3) Total RNA was extracted. 4) Syndecan-4 gene expression was verified using qPCR.

[0108] The composite extract used was a mixture of Butcher's Broom Root Extract (Maruzen Pharmaceutical Co., Ltd.) and Brown Algae Extract 1 (final concentration of each 0.1%).

[0109] The results are shown in Figure 7. As shown in FIG. 7, enhanced gene expression of syndecan-4 was observed in the combined extract of the root extract of Hagfish and brown algae extract.

[0110] Example 6 The effects of brown algae extract 2 and the composite extract on pleiotrophin gene expression were examined using the following procedure. 1) Normal human epidermal keratinocytes were seeded and cultured for 24 hours. 2) Brown algae extract 2 (final concentration 0.1%) or the composite extract was added and cultured for 6 hours. 3) Total RNA was extracted. 4) Pleiotrophin gene expression was verified using qPCR.

[0111] As a composite extract, a mixture of brown algae extract 2 and kadzu root extract (Butcher's Broom extract / Maruzen Pharmaceutical Co., Ltd.) (final concentration of each 0.1%) was used.

[0112] The results are shown in Figure 8. As shown in FIG. 8, increased pleiotrophin gene expression was observed in brown algae extract 2 and the composite extract.

[0113] Example 7 The gene expression of pleiotrophin by the composite extract was verified by the following procedure. 1) Normal human epidermal keratinocytes were seeded and cultured for 24 hours. 2) The composite extract (final concentration of each 0.1%) was added and cultured for 6 hours. 3) Total RNA was extracted. 4) Pleiotrophin (PTN) gene expression was verified using qPCR.

[0114] The composite extracts used were a mixed extract of brown algae extract 1 and kaede root extract (butcher's broom extract / Maruzen Pharmaceutical Co., Ltd.) (final concentration of both: 0.1%), and a mixed extract of brown algae extract 2 and kaede root extract (butcher's broom extract / Maruzen Pharmaceutical Co., Ltd.) (final concentration of both: 0.1%).

[0115] The results are shown in Figure 9. As shown in FIG. 9, enhanced pleiotrophin gene expression was observed in the mixed extract of brown algae extract 1 and thrush root extract, and in the mixed extract of brown algae extract 2 and thrush root extract.

[0116] Example 8 The gene expression of pleiotrophin by plant extract was verified by the following procedure. 1) Normal human epidermal keratinocytes were seeded and cultured for 24 hours. 2) Plant extract (final concentration 0.1%) was added and cultured for 6 hours. 3) Total RNA was extracted. 4) Pleiotrophin gene expression was verified using qPCR.

[0117] The plant extract used was Butcher's Broom Root Extract (Maruzen Pharmaceutical Co., Ltd.).

[0118] The results are shown in Figure 10. As shown in FIG. 10, no significant effect on pleiotrophin gene expression was observed in the Nagiikada root extract. From this, it is considered that the enhancement of pleiotrophin gene expression by the composite extract in Example 7 is due to the brown algae extract. [Industrial Applicability]

[0119] According to the present invention, a pleiotrophin receptor expression enhancer can be provided.

Claims

1. A pleiotrophin receptor expression enhancer comprising, as an active ingredient, one or more plant extracts selected from the group consisting of Ruscus aculeatus, Olea europaea, and Plantago major.

2. The pleiotrophin receptor is syndecan 4. The pleiotrophin receptor expression enhancer according to claim 1.

3. To improve skin condition, The pleiotrophin receptor expression enhancer according to claim 1 or 2.

4. Improving one or more of the following: wrinkles, sagging skin, and skin viscoelasticity and / or elasticity; The pleiotrophin receptor expression enhancer according to claim 1 or 2.

5. The present invention relates to a method for treating a flu-like disease, the method comprising administering to a patient a case in which the patient has a flu-like disease, and the patient has a flu-like disease, and the patient has a case in which ... An agent that induces degradation and / or neoplasia of the extracellular matrix.

6. The present invention relates to a method for treating a flu-like disease, the method comprising administering to a patient a case in which the patient has a flu-like disease, and the patient has a flu-like disease, and the patient has a case in which ... Skin fiber structure improver.

7. A composition for external application to the skin for enhancing pleiotrophin receptor expression, comprising as an active ingredient one or more plant extracts selected from the group consisting of Ruscus aculeatus, olive (Olea europaea), and plantain (Plantago major).

8. A composition for external application to the skin for improving skin conditions, comprising the pleiotrophin receptor expression enhancer according to claim 1 and a pleiotrophin expression enhancer.

9. A method for screening for a skin condition improving agent using the effect of enhancing the expression of pleiotrophin receptor as an index.

10. The test substance is judged to be a superior skin condition improving agent as the test substance exhibits a higher effect of enhancing the expression of pleiotrophin receptors cultured in the presence of the test substance. The screening method according to claim 9.

11. The pleiotrophin receptor is syndecan 4. The screening method according to claim 9 or 10.

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