JP2019108398A - Endo180 production promoter - Google Patents

Abstract

To provide an Endo180 production promoter among safe natural products.SOLUTION: The Endo180 production promoter comprises, as an active ingredient, at least one selected from rosmarinic acid, coffeic acid, and extract from Melissa officinalis.SELECTED DRAWING: None

Description

The present invention relates to an Endo 180 production promoter.

  In photoaging caused by long-term exposure to ultraviolet light, quantitative and qualitative changes in dermal collagen fiber structure are confirmed, and involvement in wrinkle formation is known. The collagen fiber structure is maintained in homeostasis by the metabolic balance of the synthesis and degradation of collagen fibers that occur repeatedly. This degradation process involves fragmentation of collagen by matrix metalloproteinases, uptake of fragmented collagen into fibroblasts, and degradation. Among them, involvement of a membrane protein collagen receptor “Endo 180” is known for the intracellular uptake of collagen fragments (Non-patent Document 1).

  Endo180 is a receptor also known as mannose receptor C2 (MRC2), urokinase plasminogen activator receptor related protein (uPARAP) and CD280, which induces bound ligands such as collagen to degrade in endosomes, recycling Endocytic receptors.

  Endo 180 shares homology with the macrophage mannose receptor family, the mannose receptor, phospholipase A2, and DEC-205 / MR6. Endo180 is unique among the family of mannose receptors in that it is targeted to the plasma membrane, not to the late endosomal / lysosomal compartments, but to the recycling endosomes, and for cell migration and intracellular degradation. Functions in cell motility and extracellular matrix remodeling by promoting the uptake of collagen.

  In addition, uPARAP forms a triple complex with urokinase-type plasmin activating factor (uPA) and urokinase-type plasmin activating factor receptor (uPAR), and is involved in the formation of plasmin from plasminogen.

  Endo 180 localizes in clathrin-coated pits on the cell surface and in endosomes. It is mainly expressed in fibroblasts, endothelial cells and macrophages. Tissues are mainly expressed in lung, blood vessels, bones and skin (see Patent Document 1).

  On the other hand, the dermis of the skin is also a tissue rich in collagen, but there are few reports on the involvement of Endo180 in remodeling of collagen fibers in the dermis. Heretofore, it has been known that, in photo-aged skin, fibroblasts have decreased Endo 180 expression and associated fragmented collagen in the dermal matrix (Non-patent Document 2). In addition, it is known that oxidative stress is induced in fibroblasts by fragmented collagen (Non-patent Document 3). Furthermore, it is also known that IL-1α produced by keratinocytes is involved in the mechanism of reducing Endo180 expression by ultraviolet light (Non-patent Document 4). This suggests that Endo180 may be involved in the formation of photoaged skin caused by prolonged exposure to ultraviolet light. As the mechanism, the uptake of fragmented collagen is suppressed by the decrease in the expression of Endo180 expression in fibroblasts, the regeneration of normal collagen fiber structure is impaired, and the functional failure of fibroblasts due to the accumulation of fragmented collagen around cells Therefore, it is considered that Endo 180 expression reduction may contribute to the formation of wrinkles.

  Therefore, substances that promote Endo180 expression in the skin are considered to be able to prevent or ameliorate wrinkle formation.

  Therefore, in order to enhance the promotion of Endo180 expression, for example, by promoting Endo180 expression, it is possible to promote Endo180 expression of fibroblasts in photoaging skin, promote the regeneration of normal collagen fiber structure, and suppress the formation of wrinkles. Conceivable. Caffeine, tocopherol, (-) catechin (see Patent Document 2) and the like are known as components for promoting the expression of Endo180.

Japanese Patent Application Publication No. 2012-521217 German Patent Application Publication No. 102012210384

Georgia M. et al., Journal of Cell Science, 122, 4042-48 (2009) Stefanie T. et al., Journal of Dermatological Science, 70, 42-48 (2013) GJ. Fisher et al., The American Journal of Pathology, 174, 101-114 (2009) Proceedings of the 40th Annual Meeting of the Japanese Cosmetic Association p. 16

  An object of the present invention is to find out compounds having a promoting action on Endo 180 production among compounds derived from natural products, and to provide an Endo 180 production promoting agent containing the same as an active ingredient.

  In order to solve the above problems, the Endo 180 production promoter of the present invention is characterized by blending at least one selected from rosmarinic acid, caffeic acid and Melissa extract.

  According to the present invention, by containing at least one selected from rosmarinic acid, caffeic acid and Melissa extract as an active ingredient, an Endo 180 production promoter having excellent action and effect can be provided.

Hereinafter, embodiments of the present invention will be described.
The Endo 180 production promoter of the present embodiment contains at least one selected from rosmarinic acid, caffeic acid and Melissa extract as an active ingredient.

  Rosmarinic acid has a chemical structure represented by the following structural formula (1), and can be produced by isolation and purification from a plant extract containing rosmarinic acid. Plant extracts containing such rosmarinic acid can be obtained by methods commonly used for plant extraction. As a plant containing rosmarinic acid, Melissa (Melissa officinalis L.) etc. are mentioned, for example.

  Caffeic acid has a chemical structure represented by the following structural formula (2), and can be produced by isolation and purification from a plant extract containing caffeic acid. Plant extracts containing such caffeic acid can be obtained by methods generally used for plant extraction. As a plant containing caffeic acid, for example, Melissa (Melissa officinalis L.) and the like can be mentioned.

  Melissa (Melissa officinalis L.) is a perennial herb belonging to the family Laminaceae, which is distributed all over Europe, Central Asia, North America, etc., and can be easily obtained from these areas. As a component part of Melissa which can be used as an extraction raw material, for example, an aerial part such as a leaf part, a stem part and a flower part, a root part or a mixture of these parts, etc. are mentioned, preferably a leaf part and / or a stem part It is.

  Said extract can be easily obtained by the method generally used for extraction of a plant. The extract from rosmarinic acid and caffeic acid or melissa may be an extract from rosmarinic acid and caffeic acid or melissa, a dried product obtained by drying a diluted solution of the extract, or a crude purified product thereof Or any purified product is included.

  The leaves and / or stems of Melissa, which is the raw material for extraction, can be obtained by drying and then directly or after grinding using a granulator and subjecting to solvent extraction. Drying may be performed in the sun or may be performed using a commonly used dryer. The Melissa may be used as an extraction raw material after being subjected to pretreatment such as degreasing using a nonpolar solvent such as hexane or benzene. In addition, the extraction process by the polar solvent of Melissa can be efficiently performed by pre-processing, such as a degreasing | defatting.

  As a solvent used for the said extraction, it is preferable to use water, a hydrophilic organic solvent, or these mixed solvents at the temperature below room temperature or the boiling point of a solvent.

  Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water and the like, as well as those obtained by subjecting these to various treatments. The treatment applied to water includes, for example, purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering and the like. The water which can be used as the extraction solvent also includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline and the like.

Examples of the hydrophilic organic solvent include lower alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene glycol and propylene glycol, A C2-C5 polyhydric alcohol, such as glycerol, etc. are mentioned, A mixed solvent of these hydrophilic organic solvents and water, etc. can be used.
When a mixed solvent of water and a hydrophilic organic solvent is used, 1 part by mass to 90 parts by mass with respect to 10 parts by mass of water in the case of lower alcohol, and 10 parts by mass of water in the case of lower aliphatic ketone It is preferable to add 1 mass part-40 mass parts with respect to a part. In the case of polyhydric alcohol, it is preferable to add 1 part by mass to 90 parts by mass with respect to 10 parts by mass of water.

  In the present invention, it is not necessary to adopt a special extraction method for extracting a substance having an Endo 180 production promoter from Melissa, which is an extraction raw material, and extraction using any extraction device under room temperature or reflux heating. Can.

  Specifically, the above-ground part of Melissa as an extraction raw material is charged into a treatment tank filled with an extraction solvent, and the mixture is allowed to stand for 30 minutes to 2 hours while stirring occasionally to dissolve soluble components. After filtration, the solid matter is removed by filtration, and the extractant is distilled off from the resulting extract and dried to obtain an extract. The amount of extraction solvent is usually 5 to 15 times (mass ratio) of the extraction material. Extraction conditions are about 1 to 4 hours normally at 50 degreeC-95 degreeC when water is used as an extraction solvent. Moreover, when the mixed solvent of water and ethanol is used as an extraction solvent, it is about 30 minutes-about 4 hours normally at 40 degreeC-80 degreeC. In addition, the extraction liquid obtained by extracting with a solvent can be used as Endo 180 production promoter of this invention as it is, as long as the extraction solvent has high safety.

  The extract of Melissa obtained is a diluted solution or a concentrated solution of the extract, a dried product of the extract, or a crude purified product or a purified product thereof, such as diluted, concentrated, dried, purified, etc. according to a conventional method. You may process.

  In addition, although the extract of Melissa obtained can be used as an Endo 180 production promoter, it is easier to use the concentrate or the dried product thereof. In order to obtain the dried product of the extract, a carrier such as dextrin or cyclodextrin may be added to improve the hygroscopicity. In addition, since the extract of Melissa has a peculiar odor, it is possible to carry out purification for the purpose of decolorization, deodorization, etc. within the range that does not cause a decrease in its physiological activity. When it is added to the product, it is not used in a large amount, so there is no practical problem even if it is unrefined. In addition, as refinement | purification, it can carry out by activated carbon process, adsorption resin process, ion exchange resin process etc., for example.

  The method for isolating and purifying rosmarinic acid and caffeic acid from the extract obtained as described above, the concentrate of the extract, or the dried product of the extract is not particularly limited, and may be a conventional method. It can be carried out.

  The extract of rosmarinic acid, caffeic acid and Melissa obtained as described above has an action to promote Endo 180 production, and based on these actions, it can be used as an Endo 180 production promoter of the present invention.

  In the present embodiment, at least one selected from rosmarinic acid, caffeic acid and Melissa extract may be used as the active ingredient, or these may be mixed and used as the active ingredient. When at least one or more selected from rosmarinic acid, caffeic acid and Melissa extract are used as the active ingredient, the compounding ratio is at least one selected from rosmarinic acid, caffeic acid and Melissa extract. Appropriate adjustment may be made depending on the extent of the Endo 180 production promoting action.

  The Endo 180 production-promoting agent of the present embodiment may consist only of rosmarinic acid, caffeic acid, Melissa extract or a mixture thereof, or a formulation of rosmarinic acid, caffeic acid, Melissa extract or a mixture thereof May be.

  The Endo 180 production-promoting agent of the present embodiment is a pharmaceutically acceptable carrier such as dextrin or cyclodextrin, or any other auxiliary agent, and in accordance with a conventional method, any of powders, granules, tablets, liquids, etc. Can be formulated into a dosage form of Under the present circumstances, as an adjuvant, an excipient | filler, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring agent, etc. can be used, for example. The Endo 180 production promoting agent can be used by being blended with other compositions (for example, skin external preparations, cosmetics food and drink, etc.), and can also be used as an ointment, a liquid for external use, a patch, etc. .

  When the Endo 180 production promoting agent of the present embodiment is formulated, the content of rosmarinic acid, caffeic acid, Melissa extract or a mixture thereof is not particularly limited, and may be appropriately set according to the purpose. it can.

  The Endo 180 production-promoting agent of the present embodiment is effective by blending other natural extracts having Endo 180 production-promoting activity with rosmarinic acid, caffeic acid, Melissa extract, or a mixture thereof, as necessary. It can be used as a component.

  The administration method of the Endo 180 production promoting agent of the present embodiment to a human includes transdermal administration, oral administration and the like, but a method suitable for the prevention / treatment etc. may be appropriately selected according to the type of disease. Good.

  In addition, the dose of the Endo 180 production promoting agent of the present embodiment may be appropriately increased or decreased depending on the type of disease, severity, individual differences among patients, administration method, administration period and the like.

  In addition, since the Endo 180 production promoting agent of the present embodiment has an excellent Endo 180 production promoting action, it is suitable, for example, for blending into an external preparation for skin or food and drink. In this case, rosmarinic acid, caffeic acid, Melissa extract or a mixture thereof may be blended as it is, or an Endo180 production promoting agent formulated from rosmarinic acid, caffeic acid, Melissa extract or a mixture thereof may be blended. You may

  Here, the external preparation for skin is not limited in its classification, and includes a wide range of skin cosmetics, quasi-drugs, medicines, etc. used percutaneously, and specifically, for example, ointment, Creams, milky lotions, essences, lotions, packs, foundations, lip balms, bath additives, hair tonics, hair lotions, soaps, body shampoos and the like.

  As food and drink, there is no restriction in the classification, and it includes a wide range of general food, health food, health functional food and the like which are taken orally.

  The Endo 180 production-promoting agent of the present embodiment is suitably applied to humans, but animals other than human (eg, mice, rats, hamsters, etc.) as long as the respective effects are exhibited. It can also be applied to dogs, cats, cows, pigs, monkeys, etc.).

  Hereinafter, the present invention will be specifically described by showing test examples, but the present invention is not limited to the following examples. In the following test examples, the products (sample Nos. 1 to 3) shown in Table 1 were used as test samples.

[Table 1]
Sample No. Product Name Manufacturer Name / Seller Name
1 Rosmarinic acid Wako Pure Chemical Industries Ltd. 2 caffeic acid Wako Pure Chemical Industries Ltd. 3 Melissa extract BG-J freeze-dried product Maruzen Pharmaceutical Co., Ltd.

[Test Example 1]
-Test for promoting Endo 180 production in skin fibroblasts-
Human normal skin fibroblasts (NB1 RGB) were cultured using Dulbecco's MEM containing 10% FBS, and then cells were harvested by trypsin treatment. The collected cells were diluted with Dulbecco's MEM containing 0.25% FBS to a concentration of 1.0 × 10 ⁵ cells / mL, seeded at 2 mL per well in a 6-well microplate, and cultured overnight. After completion of the culture, the medium was removed, and 2 mL of a test sample dissolved in Dulbecco's MEM containing 0.25% FBS was added to each well and cultured for 24 hours. After completion of the culture, total protein was prepared by a conventional method.

  After SDS-PAGE was performed on the prepared sample, Western blotting was performed using anti-Endo 180 antibody and anti-GAPDH antibody. The protein production amount of Endo 180 is obtained by dividing the Net intensity of Bands derived from Endo 180 in the protein prepared from each of the cells cultured with and without the test sample, by the Net intensity of the band derived from GAPDH, The correction value per cell was calculated. The calculation method of Endo 180 production promotion rate is as follows.

Endo 180 production promotion rate (%) = A / B x 100
A: Endo 180 production per cell upon addition of test sample
B: Endo 180 production per cell when no test sample is added

[Table 2] ENDO 180 production promotion rate (%)
Sample concentration (μg / mL)
1 10
Sample No. 119.4% 114.3%
Sample No. 2 129.9% 174.6%

Sample concentration (μg / mL)
5 50
Sample No. 3 105.4% 127.0%

  The Endo 180 production promoter containing at least one or more selected from rosmarinic acid, caffeic acid and Melissa extract of the present invention as an active ingredient can greatly contribute to the prevention or amelioration of wrinkle formation.

Claims (1)

An Endo 180 production promoter for promoting the uptake of fragmented collagen into fibroblasts, which comprises caffeic acid as an active ingredient.