Skin function improving agent, neurotrophic factor production promoting agent, CXCL1 production inhibitor, TGFB3 production promoting agent

Abstract

The present invention aims to provide a skin function improving agent, a neurotrophic factor production promoting agent, a CXCL1 production inhibitor, and a TGFB3 production promoting agent containing peony extract. [Solution] The present invention provides a skin function improving agent, a neurotrophic factor production promoting agent, a CXCL1 production inhibitor, and a TGFB3 production promoting agent containing peony extract.

Description

Technical Field

[0001] The present invention relates to a skin function improver, a nerve growth factor production promoter, a CXCL1 production inhibitor, and a TGFB3 production promoter containing peony extract.

Background Art

[0002] Schwann cells, which are one of the glial cells of the peripheral nervous system, play a major role in the maintenance and regeneration of nerves when damaged. It is known that nerve growth factors such as BDNF (Brain derived neurotrophic factor), GDNF (Glial cell line derived neurotrophic factor), and NTF3 (Neurotrophin 3) are produced from Schwann cells (Non-Patent Document 1). In addition, Schwann cells secrete inflammatory factors when nerves are damaged. Among them, CXCL1 (C-X-C motif chemokine ligand 1) has been reported to inhibit axonal growth of nerve cells (Non-Patent Document 2), and it has also been reported that the secretion amount of inflammatory factors increases with aging (Non-Patent Document 3). Furthermore, it is also known that TGFB3 (Transforming growth factor beta 3) secreted by Schwann cells promotes wound healing (Non-Patent Document 4). Based on the above, it is expected that the functions of Schwann cells can enhance the barrier function, repair function, and immune function of the skin, and research and development are underway.

[0003] Peony (Paeonia lactiflora Pallas) is a crude drug formulated in many traditional Chinese medicine preparations including Shaoyao Gancao Decoction, and its effect on the skin is expected and it is also applied to cosmetics (Patent Document 1). However, the action of peony on Schwann cells has not been elucidated.

Prior Art Documents

Non-Patent Documents

[0004] [Non-Patent Document 1] Eric R. Bray, et al, Schwann cells as underestimated, major players in human skin physiology and pathology, Experimental Dermatology, 16 November 2019, https: / / doi.org / 10.1111 / exd.14060 [Non-Patent Document 2] Antonia Teona Deftu, et al, CXCL1 and CXCL2 Inhibit the Axon Outgrowth in a Time- and Cell-Type-Dependent Manner in Adult Rat Dorsal Root Ganglia Neurons, NeurochemRes.2019 Sep;44(9):2215-2229 [Non-Patent Document 3] Andres Fuentes-Flores, et al, Senescent Schwann cells induced by aging and chronic denervation impair axonal regeneration following peripheral nerve injury, EMBO Mol Med.2023 Dec;15(12):e17907. [Non-Patent Document 4] Min-Yi Ou, et al, Dedifferentiated Schwann cell-derived TGF-β3 is essential for the neural system to promote wound healing, Theranostics.2022;12(12):5470-5487. [Patent Documents]

[0005] [Patent Document 1] Japanese Patent Publication No. 2023-41502 [Overview of the project] [Problems that the invention aims to solve]

[0006] The present invention aims to provide a skin function improving agent, a neurotrophic factor production promoting agent, a CXCL1 production inhibitor, and a TGFB3 production promoting agent containing peony extract. [Means for solving the problem]

[0007] The means for solving the problems of the present invention are as follows. (1) A skin function improving agent containing peony extract. (2) A neurotrophic factor production promoter containing peony extract. (3) The agent according to (2), wherein the neurotrophic factor is one or more selected from BDNF, GDNF, and NTF3. (4) A CXCL1 production inhibitor containing peony extract. (5) A TGFB3 production promoter containing peony extract. (6) The agent described in one or more of (2) to (5) in Schwann cells. [Effects of the Invention]

[0008] The peony extract of the present invention exhibits effects that improve skin function, promote neurotrophic factor production, inhibit CXCL1 production, and promote TGFB3 production. [Modes for carrying out the invention]

[0009] The following describes embodiments for carrying out the present invention.

[0010] BDNF is the abbreviation of Brain derived neurotrophic factor, which is a kind of neurotrophic factor and is called brain-derived neurotrophic factor. It is known to promote the maintenance, growth, and differentiation of nerve cells.

[0011] GDNF is the abbreviation of Glial cell line derived neurotrophic factor, which is a kind of neurotrophic factor and is called glial cell line-derived neurotrophic factor. It is known to promote the maintenance, growth, and differentiation of nerve cells.

[0012] NTF3 is the abbreviation of Neurotrophin 3, which is a kind of neurotrophic factor. It is known to promote the maintenance, growth, and differentiation of nerve cells.

[0013] CXCL1 is the abbreviation of C-X-C motif chemokine ligand 1, which is an inflammatory factor that inhibits axonal elongation of nerve cells during nerve injury. It is known to increase with aging.

[0014] TGFB3 is the abbreviation of Transforming growth factor beta 3, which is a kind of cytokine. It is known to promote wound healing.

[0015] [Paeonia lactiflora Pall.] The Paeonia lactiflora Pallas used in the Paeonia lactiflora extract used in the present invention is a perennial herbaceous plant classified in the genus Paeonia of the family Paeoniaceae.

[0016] Examples of the constituent parts of Paeonia lactiflora that can be used for extraction include roots, leaves, stems, flowers, buds, bark, whole above-ground herbs, etc., but roots are preferably used in terms of effectiveness.

[0017] In this invention, peony can be used for extraction in its raw state, but considering the extraction efficiency, it is preferable to perform extraction after processing such as finely chopping, drying, or grinding. Extraction is carried out by immersion in the extraction solvent. Stirring or homogenization in the extraction solvent may be performed to increase the extraction efficiency. The extraction temperature should be between approximately 5°C and below the boiling point of the extraction solvent. The extraction time will vary depending on the type of extraction solvent and the extraction temperature, but it should be between approximately 4 hours and 30 days.

[0018] As the extraction solvent, polar organic solvents such as lower alcohols like methanol, ethanol, propanol, and isopropanol; polyhydric alcohols like 1,3-butylene glycol, propylene glycol, dipropylene glycol, and glycerin; ethers like ethyl ether and propyl ether; esters like ethyl acetate and butyl acetate; and ketones like acetone and ethyl methyl ketone can be used, and one or more of these can be selected and used. Physiological saline, phosphate buffer, or phosphate-buffered saline may also be used. An aqueous solution of ethanol is preferred as the extraction solvent, with a 50% by volume aqueous solution being the most preferred.

[0019] The peony extract prepared with the above solvent can be used as is, but it may also be concentrated and dried and then redissolved in water or a polar solvent, or purified by decolorization, deodorization, desalting, etc., to the extent that it does not impair the skin physiological function-enhancing effects, or fractionated by column chromatography before use. For preservation, it can also be freeze-dried after purification and dissolved in a solvent when needed. It can also be encapsulated in vesicles such as liposomes or microcapsules for use.

[0020] The peony extract used in this invention may be extracted directly from the plant or be a commercially available product. Examples of commercially available peony extracts include Peony Extract-BG, Peony Extract-ET (both manufactured by Yamada Yakken Co., Ltd.), Peony Liquid, Falcorex Peony B (both manufactured by Ichimaru Falcos Co., Ltd.), Peony Extract BG-50, Peony Extract ET-30 (both manufactured by Koei Kogyo Co., Ltd.), Peony Extract-J, Peony Extract BG-JC (both manufactured by Maruzen Pharmaceutical Co., Ltd.). In this invention, it is preferable to use an extract obtained from peonies organically grown in Yunosawa, Mashike-cho, Hokkaido.

[0021] The following are the growing conditions for organic cultivation in Yunozawa, Mashike Town, Hokkaido.

[0022] [Hair thickening] Mashike is located in the south of the Rumoi Subprefecture in northwestern Hokkaido, and includes the Shokanbetsu-dake mountain, which has an elevation of 1492m. In this invention, it is preferable to cultivate the crop in the Yunosawa district, which is at a higher elevation within Mashike.

[0023] [Field preparation] Before planting, organic fertilizer is applied to the field to improve the soil. Specifically, one or more organic fertilizers selected from fermented chicken manure, oil cake, fermented oil cake, bone meal, fish meal, rice bran, fermented rice bran, leaf mold, bark compost, magnesium lime, slaked lime, calcium carbonate, and guano are used in combination.

[0024] Fermented chicken manure, oilseed meal, fermented oilseed meal, bone meal, fish meal, rice bran, fermented rice bran, leaf mold, bark compost, and iodine-based phosphate fertilizer are mainly used to supply the three major nutrients: nitrogen, phosphorus, and potassium. These can be used individually or in combination of two or more. The amount of these fertilizers applied can be increased or decreased depending on the original soil condition.

[0025] Fertilization should ideally be carried out at least 7 days before transplanting. Applying fertilizer within 6 days of transplanting may result in insufficient soil improvement, leading to fertilizer burn or poor initial growth. Organic fertilizers can be applied in furrows or as a whole, but whole-field application is preferable for efficiency.

[0026] [Planting] The plants harvested in September and October were divided and planted directly in the ground.

[0027] [Training] Applying fertilizer as needed will promote more significant growth. Remove weeds as needed and water according to the dry conditions.

[0028] [harvest] The rhizomes are harvested in September or October.

[0029] Each preparation containing the peony extract of the present invention can be used alone, but it can also be incorporated into various compositions such as pharmaceuticals, quasi-drugs, foods and beverages, and cosmetics.

[0030] The preparations containing the peony extract of the present invention are not limited in any way in terms of their form or the presence or absence of other components. Regarding form, any form such as liquid, paste, gel, solid, or powder can be selected according to its intended use, and may optionally contain the necessary vehicles (excipients), solvents, and other general additives (antioxidants, colorants, dispersants, etc.) to achieve that form.

[0031] The compositions containing the peony extract of the present invention can take any dosage form. When the composition is a skin cosmetic, hair cosmetic, cleanser, etc., it can be provided as a solubilizing system such as a lotion, a dispersion system such as a calamine lotion, or an emulsifying system such as a cream or emulsion. Furthermore, it can be provided in various dosage forms such as an aerosol form filled with a propellant, an ointment, or a poultice.

[0032] Specifically, examples include various cosmetics such as emulsions, creams, lotions, toners, masks, serums, cleansers, and makeup cosmetics; cosmetics in various forms such as liquids, ointments, powders, granules, aerosols, patches, and poultices; quasi-drugs and topical medicines.

[0033] Furthermore, in the case of oral pharmaceuticals, they can be in the form of liquid preparations such as drinks and intravenous solutions, solid preparations such as gum and candy, capsules, powders, granules, and tablets—common dosage forms.

[0034] In addition to the above, compositions containing each agent containing the peony extract of the present invention may also contain, depending on the intended use and requirements, any ingredients commonly used in pharmaceuticals, quasi-drugs, skin cosmetics, hair cosmetics, cleansers, and oral pharmaceuticals, such as water, oily components, humectants, powders, pigments, emulsifiers, solubilizers, gelling agents, cleansing agents, UV absorbers, anti-inflammatory agents, thickeners, pH adjusters, chelating agents, drugs (pharmaceutical components), fragrances, resins, antibacterial and antifungal agents, antioxidants, alcohols, etc. Furthermore, within the limits that do not impair the effects of the present invention, it is also possible to use other humectants, anti-aging agents, whitening agents, antioxidants, and slimming agents, or various plants / fungi or their extracts. [Examples]

[0035] The present invention will be specifically described below with reference to examples, but this will not limit the scope of the present invention.

[0036] First, the method for preparing the extracts used in the sample is shown.

[0037] [Peony extract] (1) From cultivation to harvesting Peonies were cultivated in an organic JAS-certified field in Yunosawa, Mashike Town, Hokkaido. In mid-September, one week before transplanting, the field was prepared by applying bark compost, organic chicken manure, organic rice bran, organic lime, organic guano, organic rapeseed oil cake, and calcium carbonate. The plants were divided so that the rows were spaced 90-140 cm apart and the plants 30-60 cm apart, and then transplanted. Weeds were removed as needed, and watering was performed according to the dry conditions. The rhizomes were harvested in September or October, four years after planting, and washed with water or hot water. For storage, they were kept under dry conditions. They were also preserved by freeze-drying. (2) Preparation of peony extract The rhizomes of the peony obtained in (1) were dried and immersed in 20 times the mass of a 50% ethanol aqueous solution. The liquid after filtration was collected and the solvent was removed by distillation. The obtained extract was dissolved in a 50% 1,3-butylene glycol aqueous solution to a concentration of 0.5% by mass of extract, and then filtered again to prepare the peony extract.

[0038] [Confirmation of gene expression changes using human-derived Schwann cells] Human-derived Schwann cells 3 × 10 5 Cells were seeded in 6-well plates at a density of cells per well and cultured overnight in DMEM medium containing 5% FBS. The medium was then replaced with 100 μg / mL of peony extract, and the cells were cultured for 24 hours at 37°C in a 5% CO2 incubator. RNA was extracted from the collected cells using a commercially available RNA extraction kit (Quick Gene RNA Cultured Cell Kit S), and after cDNA synthesis, gene expression was confirmed by real-time PCR using the CyberGreen method with the primers listed in Table 1. GAPDH was used as an internal standard. mRNA expression levels are shown as relative values, with the expression level without peony extract set to 1. The results are shown in Table 2.

[0039] [Table 1]

[0040] [Table 2]

[0041] As shown in Table 2, the addition of peony extract significantly increased the expression of neurotrophic factors BDNF, GDNF, and NTF3. Furthermore, the expression of CXCL1, which inhibits axonal elongation of nerve cells, significantly decreased. In addition, the expression of TGFB3, which promotes wound healing, significantly increased.

[0042] Based on the above, the peony extract of the present invention exhibits excellent effects in promoting neurotrophic factor production, inhibiting CXCL1 production, and promoting TGFB3 production. Furthermore, by exhibiting these effects, it improves the skin's barrier function, repair function, and immune function, thereby exhibiting excellent skin function improvement effects.

Claims

1. A skin function improving agent containing peony extract.

2. A neurotrophic factor production promoter containing peony extract.

3. The agent according to claim 2, wherein the neurotrophic factor is one or more selected from BDNF, GDNF, and NTF3.

4. A CXCL1 production inhibitor containing peony extract.

5. A TGFB3 production promoter containing peony extract.

6. The agent according to one or more of claims 2 to 5 in Schwann cells.

Images