JP2017128538A - Endo180 production promoter - Google Patents

Abstract

PROBLEM TO BE SOLVED: 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 Endo180 production promoter.

  In photoaging caused by long-term exposure to ultraviolet rays, quantitative and qualitative changes in the dermal collagen fiber structure have been confirmed, and their involvement in wrinkle formation is known. The collagen fiber structure is maintained in homeostasis by the metabolic balance between repetitive collagen fiber synthesis and degradation. This degradation process includes fragmentation of collagen by matrix metalloprotease and incorporation of fragmented collagen into fibroblasts and degradation. Among these, the collagen receptor “Endo180”, which is a membrane protein, is known to be involved in the incorporation of collagen fragments into cells (Non-patent Document 1).

  Endo180 is a receptor also known as mannose receptor C2 (MRC2), urokinase plasminogen activator receptor associated protein (uPARAP) and CD280, which induces a bound ligand, such as collagen, for degradation in the endosome. Is an endocytic receptor.

  Endo180 shares homology with the mannose receptor, phospholipase A2, and DEC-205 / MR6, a family of macrophage mannose receptors. Endo180 is unique among the family of mannose receptors in that it is targeted to the recycling endosome rather than the plasma membrane to the late endosomal / lysosomal compartment for cell migration and intracellular degradation. It functions in cell motility and extracellular matrix remodeling by promoting the uptake of collagen.

  In addition, uPARAP forms a ternary complex with urokinase-type plasmin activator (uPA) and urokinase-type plasmin activator receptor (uPAR), and is involved in the production of plasmin from plasminogen.

  Endo180 is localized in clathrin-covered pits and endosomes on the cell surface. It is mainly expressed in fibroblasts, endothelial cells, and macrophages. As a tissue, it is mainly expressed in lung, blood vessel, bone and skin (see Patent Document 1).

  On the other hand, although the dermis of the skin is also a tissue rich in collagen, there are few reports regarding the involvement of Endo180 in remodeling of collagen fibers in the dermis. So far, it has been known that, in photoaged skin, the expression of Endo180 in fibroblasts is reduced and fragmented collagen is accumulated in the dermal matrix (Non-Patent Document 2). Further, 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 lowering Endo180 expression by ultraviolet light (Non-patent Document 4). This suggests that Endo180 is involved in the formation of photoaged skin caused by long-term exposure to ultraviolet rays. As the mechanism, the reduction of Endo180 expression in fibroblasts suppresses the uptake of fragmented collagen, thereby impairing the regeneration of normal collagen fiber structure, and the impairment of fibroblasts due to the accumulation of fragmented collagen around the cells. Therefore, the decrease in Endo180 expression may contribute to the formation of wrinkles.

  Therefore, it is considered that a substance that promotes Endo180 expression in the skin can prevent or improve wrinkle formation.

  Therefore, in order to enhance the promotion of Endo180 expression, for example, by promoting Endo180 expression, it promotes Endo180 expression of fibroblasts in photoaged skin, promotes regeneration of normal collagen fiber structure, and suppresses the formation of wrinkles. Conceivable. As components that promote the expression of Endo180, caffeine, tocopherol, (−) catechin (see Patent Document 2), and the like are known.

Special table 2012-521217 gazette German Patent Application Publication No. 102012122384

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) Abstract of the 40th Annual Meeting of the Japan Cosmetic Science Society p.16

  An object of the present invention is to find an endo180 production promoter from natural products and to provide an endo180 production promoter containing the compound as an active ingredient.

  In order to solve the above problems, the Endo180 production promoter of the present invention is characterized by blending at least one selected from rosmarinic acid, caffeic acid and Melissa extract.

  ADVANTAGE OF THE INVENTION According to this invention, the Endo180 production promoter excellent in the effect can be provided by containing as an active ingredient at least 1 or more types chosen from a rosmarinic acid, a caffeic acid, and a Melissa extract.

Embodiments of the present invention will be described below.
The Endo180 production promoter of this 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. Such a plant extract containing rosmarinic acid can be obtained by a method generally used for plant extraction. Examples of plants containing rosmarinic acid include Melissa (Melissa officinalis L.).

  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. Such a plant extract containing caffeic acid can be obtained by a method generally used for plant extraction. Examples of plants containing caffeic acid include Melissa officinalis L. and the like.

  Melissa (Melissa officinalis L.) is a perennial herb that is distributed throughout Europe, Central Asia, North America, etc., and can be easily obtained from these regions. Examples of the constituent parts of Melissa that can be used as an extraction raw material include above-ground parts such as leaves, stems, and flower parts, root parts, or a mixture of these parts, but preferably leaves and / or stems. It is.

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

  The leaf part and / or stem part of Melissa, which is the raw material for extraction, can be obtained by drying or pulverizing it as it is or using a crusher and subjecting it to solvent extraction. Drying may be performed in the sun or using a commonly used dryer. The melissa may be used as an extraction raw material after pretreatment such as degreasing with a nonpolar solvent such as hexane or benzene. In addition, by performing pretreatments such as degreasing, extraction with Melissa's polar solvent can be performed efficiently.

  As the solvent used for the extraction, it is preferable to use water, a hydrophilic organic solvent, or a mixed solvent thereof at a temperature from room temperature to the boiling point of the 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 water that has been subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, and the like. The water that can be used as the extraction solvent includes purified water, hot water, ion exchange 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, propylene glycol, Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycerin, and a mixed solvent of these hydrophilic organic solvents and water can be used.
In addition, when using the mixed solvent of the said water and a hydrophilic organic solvent, in the case of a lower alcohol, 1 mass part-90 mass parts with respect to 10 mass parts of water, in the case of a lower aliphatic ketone, 10 masses of water. It is preferable to add 1 to 40 parts by mass with respect to parts. 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 Endo180 production promoter from Melissa, which is an extraction raw material, and extraction is performed using an arbitrary extraction device at room temperature or under reflux heating. Can do.

  Specifically, the above-ground part of Melissa as an extraction raw material is put into a processing tank filled with the extraction solvent, and further, if necessary, it is allowed to stand for 30 minutes to 2 hours to elute soluble components. Then, the solid is removed by filtration, the extraction solvent is distilled off from the obtained extract, and the extract is obtained by drying. The amount of the extraction solvent is usually 5 to 15 times (mass ratio) of the extraction raw material. The extraction conditions are usually about 1 to 4 hours at 50 to 95 ° C. when water is used as the extraction solvent. Moreover, when the mixed solvent of water and ethanol is used as an extraction solvent, it is normally 30 minutes-about 4 hours at 40 to 80 degreeC. In addition, the extraction liquid obtained by extracting with a solvent can be used as it is as an Endo180 production promoter of the present invention as long as the extraction solvent is highly safe.

  The resulting Melissa extract is diluted, concentrated, dried, purified, etc. according to conventional methods to obtain a diluted or concentrated solution of the extract, a dried product of the extract, or a crudely purified product or purified product thereof. Processing may be performed.

  The obtained Melissa extract can be used as an Endo180 production promoter, but a concentrated solution or a dried product thereof is easier to use. In obtaining a dried extract, a carrier such as dextrin or cyclodextrin may be added to improve hygroscopicity. In addition, since the Melissa extract has a unique odor, it can be purified for the purpose of decolorization, deodorization, etc. within a range that does not cause a decrease in its physiological activity. When added to the product, it is not used in large quantities, so there is no practical problem even if it is not purified. The purification can be performed, for example, by activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, or the like.

  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 is a conventional method. It can be carried out.

  The extracts of rosmarinic acid, caffeic acid and Melissa obtained as described above have an Endo180 production promoting action, and can be used as the Endo180 production promoting agent of the present invention based on these actions.

  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 selected from rosmarinic acid, caffeic acid and Melissa extract is used as the active ingredient, the blending ratio thereof is at least one selected from rosmarinic acid, caffeic acid and Melissa extract. What is necessary is just to adjust suitably according to the grade etc. of Endo180 production promotion effect.

  The Endo180 production promoting agent of the present embodiment may consist 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. But you can.

  The Endo180 production promoting agent of the present embodiment is an arbitrary powdery, granular, tablet, liquid, etc., using a pharmaceutically acceptable carrier such as dextrin and cyclodextrin and other optional auxiliaries according to a conventional method. It can be formulated into a dosage form. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring / flavoring agent, and the like can be used. The Endo180 production promoting agent can be used by blending with other compositions (for example, external preparations for skin, cosmetic foods and drinks, etc.), and can also be used as an ointment, a liquid for external use, a patch, and the like. .

  When the Endo180 production promoting agent of this 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.

  In addition, the Endo180 production promoting agent of the present embodiment is effective by blending other natural extracts having an Endo180 production promoting action together with rosmarinic acid, caffeic acid, Melissa extract, or a mixture thereof, if necessary. It can be used as a component.

  Examples of the method for administering the Endo180 production-promoting agent of this embodiment to humans include transdermal administration, oral administration, and the like. Depending on the type of disease, a suitable method for its prevention / treatment may be selected as appropriate. Good.

  In addition, the dose of the Endo180 production promoting agent of this embodiment may be appropriately increased or decreased depending on the disease type, severity, individual differences among patients, administration method, administration period, and the like.

  Moreover, since the Endo180 production promotion agent of this embodiment has the outstanding Endo180 production promotion effect | action, it is suitable for mix | blending with a skin external preparation or food-drinks, for example. In this case, rosmarinic acid, caffeic acid, Melissa extract or a mixture thereof may be blended as they are, or an Endo180 production promoting agent formulated from rosmarinic acid, caffeic acid, Melissa extract or a mixture thereof is blended. May be.

  Here, the topical skin preparation is not limited in its category, and includes a wide range of skin cosmetics, quasi-drugs, pharmaceuticals, and the like used transdermally. Creams, milky lotions, beauty essences, lotions, packs, foundations, lip balms, bath salts, hair nicks, hair lotions, soaps, body shampoos and the like.

  There is no restriction on the category of food and drink, and it includes a wide range of foods such as general foods, health foods, and functional health foods that are taken orally.

  The Endo180 production promoting agent of the present embodiment is suitably applied to humans, but as long as each effect is exhibited, animals other than humans (for example, mice, rats, hamsters, It can also be applied to dogs, cats, cows, pigs, monkeys, etc.).

  Hereinafter, although a test example is shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all. In the following test examples, 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 Lyophilized Product Maruzen Pharmaceutical Co., Ltd.

[Test Example 1]
-Endo180 production promoting action test in skin fibroblasts-
Human normal skin fibroblasts (NB1RGB) were cultured using Dulbecco MEM containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with Dulbecco MEM containing 0.25% FBS to a concentration of 1.0 × 10 ⁵ cells / mL, then seeded at 2 mL per well on 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.

  SDS-PAGE was performed on the prepared sample, and then Western blotting was performed using an anti-Endo180 antibody and an anti-GAPDH antibody. The protein production amount of Endo180 is obtained by dividing the net intensity (band intensity) of the band derived from Endo180 in the protein prepared from each of the cells cultured with and without the addition of the test sample by the net intensity of the band derived from GAPDH, A correction value per cell was calculated. The calculation method of Endo180 production promotion rate is as follows.

Endo180 production promotion rate (%) = A / B × 100
A: Endo180 production amount per cell at the time of test sample addition
B: Endo180 production amount per cell when no test sample is added

[Table 2] Endo180 production promotion rate (%)
Sample concentration (μg / mL)
1 10
Sample No. 1 119.4% 114.3%
Sample No. 2 129.9% 174.6%

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

  The Endo180 production promoter containing at least one selected from rosmarinic acid, caffeic acid and Melissa extract of the present invention as an active ingredient can greatly contribute to the prevention or improvement of wrinkle formation.

Claims (1)

An Endo180 production promoter comprising at least one selected from rosmarinic acid, caffeic acid and Melissa extract as an active ingredient.