JP2019182753A - Endo180 production promoter - Google Patents

Abstract

To discover, among natural product-derived components, one having an excellent Endo180 production promotion action, and provide an Endo180 production promoter containing the same as an active ingredient.SOLUTION: The present invention provides an Endo180 production promoter containing, as an active ingredient, one or more selected from the group consisting of lotus germ extract, Alpinia speciosa leaf extract and Lactobacillus plantarum.SELECTED DRAWING: None

Description

  The present invention relates to an Endo180 production promoter containing a natural product-derived component as an active ingredient.

  In photoaging caused by long-term exposure to ultraviolet rays, quantitative and qualitative changes in the dermal collagen fiber structure have been confirmed, and it is considered to be involved in the formation of wrinkles. The collagen fiber structure is maintained in homeostasis by metabolic balance due to repeated synthesis and degradation of collagen fibers. In the degradation process, fragmentation of collagen by matrix metalloprotease and uptake and degradation of the fragmented collagen into fibroblasts occur. Involvement of collagen receptor “Endo180”, which is a protein, is known (Non-patent Document 1).

  Endo180, also known as mannose receptor C2 (MRC2), urokinase plasminogen activator receptor associated protein (uPARAP) and CD280, is a re-used endocytosis receptor that binds to ligands such as collagen This is taken up into cells and induced to degrade in endosomes.

  Endo180 has homology with the mannose receptor, phospholipase A2, DEC-205 / MR6, and the like, which are the macrophage mannose receptor family. Endo180 is unique among the family of mannose receptors in that it is targeted to the recycling endosome rather than the cell membrane to the late endosomal / lysosomal compartment, and is a collagen for cell migration and intracellular degradation. It promotes cell uptake and functions in cell motility and extracellular matrix remodeling. 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 pits covered with clathrin on the cell surface, endosomes, and the like, and is mainly expressed in fibroblasts, endothelial cells, and macrophages. As a tissue, it is mainly expressed in lungs, blood vessels, bones 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 Endo180 expression is decreased in fibroblasts in photoaged skin, and fragmented collagen is accumulated in the dermal matrix accordingly (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). From these facts, Endo180 is considered to be involved in the formation of photoaged skin (for example, the formation of wrinkles) caused by long-term exposure to ultraviolet rays. As the mechanism, when the expression of Endo180 in fibroblasts decreases, the uptake of fragmented collagen is suppressed, so that the regeneration of normal collagen fiber structure is inhibited, and the fragmented collagen accumulates around the cells. It is thought to induce dysfunction of fibroblasts.

  Therefore, for example, if Endo180 expression can be promoted in fibroblasts, it promotes reuse of fragmented collagen by metabolism and regeneration of normal collagen fiber structure, and suppresses the formation of photoaged skin and wrinkles due to this. It is thought that you can.

Special table 2012-521217 gazette

J. Cell Sci., 2009, vol.122, pp.4042-4048 J. Dermatol. Sci., 2013, vol.70, pp.42-48 Am. J. Pathol., 2009, vo.174, pp.101-114 Abstract from the 40th Annual Meeting of the Japan Cosmetic Science Society, 2015, p.16

  An object of the present invention is to find an endo180 production promoting agent having an excellent endo180 production promoting action from among highly safe natural product-derived components, and to provide an endo180 production promoting agent comprising the same.

  In order to solve the above-mentioned problems, the Endo180 production promoter of the present invention comprises, as an active ingredient, one or more selected from the group consisting of lotus germ extract, moon peach leaf extract, and Lactobacillus plantarum It is characterized by.

  According to the present invention, by using one or more selected from the group consisting of a lotus germ extract derived from a natural product, a moon peach leaf extract, and a Lactobacillus plantarum as an active ingredient, the effect is excellent. And, Endo180 production promoter with high safety can be provided.

Embodiments of the present invention will be described below.
The Endo180 production promoter of this embodiment has one or more selected from the group consisting of a lotus germ extract, a moon peach leaf extract, and a Lactobacillus plantarum as an active ingredient.

[Lotus germ extract, moon peach leaf extract]
The lotus germ extract and moon peach leaf extract used in this embodiment are extracts obtained from lotus (scientific name: Nelumbo nucifera Gaertn.) And moon peach (scientific name: Alpinia zerumbet or Alpinia speciosa (Wendl.) K. Schum.). It is.

  Here, the “extract” in the present embodiment includes an extract obtained using the above natural product as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or these Both of these crudely purified products and purified products are included.

  The lotus (Nelumbo nucifera Gaertn.) Is a perennial aquatic plant belonging to the genus Lotusaceae native to tropical Asia. In Japan, lotus root is eaten as a lotus root in Japan and is a highly safe plant. Lotus is distributed in southeastern Europe, northern Australia, eastern Asia, etc., and is cultivated in ponds, paddy fields, moats, etc., and can be easily obtained from these areas. The constituent part of the lotus used as an extraction raw material is germ. The lotus germ is a green, rod-shaped germ in a lotus seed.

  Tsuki Peach (Alpinia zerumbet or Alpinia speciosa (Wendl.) K. Schum.) Is a perennial evergreen herb belonging to the genus Glyceraceae and is distributed from southern Kyushu to India and is easily obtained from these regions be able to. Tsuki-Peach is called Sannin in Okinawa and is used in Muchy, a traditional confectionery since the Ryukyu dynasty. The part used as an extraction raw material is a leaf part.

  The extract from the natural product can be obtained by drying the extraction raw material, pulverizing it as it is or using a crusher, and subjecting it to extraction with an extraction solvent. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing a pretreatment such as degreasing, the extraction material can be efficiently extracted with a polar solvent.

  As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water and hydrophilic organic solvents. These are used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. It is preferable.

  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, and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in this embodiment includes purified water, hot water, ion exchange water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic 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. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when a mixed liquid of water and lower aliphatic alcohol is used as an extraction solvent, the mixing ratio of water and lower aliphatic alcohol is preferably 9: 1 to 1: 9 (volume ratio), More preferably, it is 7: 3 to 2: 8 (capacity ratio). In addition, when a mixed liquid of water and a lower aliphatic ketone is used, the mixing ratio of water and the lower aliphatic ketone is preferably 9: 1 to 2: 8 (volume ratio). When using the liquid mixture with a monohydric alcohol, it is preferable that the mixing ratio of water and a polyhydric alcohol is 8: 2 to 1: 9 (volume ratio).

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent 5 to 15 times (mass ratio) of the extraction raw material, the soluble components are extracted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. When the solvent is distilled off from the obtained extract, a paste-like concentrate is obtained, and when this concentrate is further dried, a dried product is obtained.

  Since the lotus germ extract and moon peach leaf extract obtained as described above have an excellent Endo180 production promoting action, they can be used as an active ingredient of an Endo180 production promoter.

[Lactobacillus plantarum]
In this embodiment, Lactobacillus plantarum can also be used. Lactobacillus plantarum is a kind of lactic acid bacteria, but it is included in plant lactic acid bacteria because it is mainly isolated from vegetable fermented foods such as vegetable pickles, kimchi, miso, etc., producing fermented milk such as yogurt and lactic acid bacteria beverages It is different from animal lactic acid bacteria used in

  In this embodiment, any lactic acid bacteria classified into Lactobacillus plantarum may be used, but since the effects described below are preferably exhibited, Lactobacillus plantarum 22A-1 (FERM P-21409) ), Lactobacillus plantarum 22A-3 (FERM P-21411), and Lactobacillus plantarum 22B-2 (FERM P-21410). Talam 22A-3 (FERM P-21411) is particularly preferable.

  The three strains of Lactobacillus plantarum 22A-1 (FERM P-21409), Lactobacillus plantarum 22A-3 (FERM P-21411), and Lactobacillus plantarum 22B-2 (FERM P-21410) are: It is a strain isolated and identified from pickles by the present inventors and deposited with the Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology.

In this embodiment, a large amount can be obtained by growing the Lactobacillus plantarum by culturing. The medium for culturing the Lactobacillus plantarum is not particularly limited, and a medium usually used for culturing lactic acid bacteria or a modified medium thereof can be used. Examples of medium components to be mixed in such a medium include carbon sources such as glucose; nitrogen sources such as yeast extract, meat extract, casein, and protein hydrolysates thereof; sodium chloride, sodium phosphate, and phosphoric acid. Inorganic salts such as potassium, ammonium citrate, and magnesium sulfate; and components suitable for the growth of lactic acid bacteria, such as vitamins such as thiamine and inositol. Moreover, you may use the culture medium containing the plant material etc. which can be utilized as a foodstuff raw material.
In the present embodiment, it is not necessary to use a component derived from animal milk as a medium component. When ingredients derived from animal milk are used as medium components, they may cause milk allergy, which may limit the use by allergic patients. In addition, culturing lactic acid bacteria produces undesirable aromas. In some cases, it may not be suitable as a cosmetic ingredient.

  In the culture of Lactobacillus plantarum, the culture conditions such as the form of the medium, pH, culture temperature, culture time, culture method, etc. are not particularly limited, and conventionally known conditions can be employed. For example, the medium may be a liquid medium or a solid medium, and the culture temperature is preferably 25 to 45 ° C., more preferably 30 to 37 ° C., and the culture time is preferably 4 to 74 hours, more preferably It can be 18 to 48 hours. Further, in the case of a liquid medium, examples of the culture method include standing, stirring, shaking, aeration, etc., any of which may be adopted.

  The obtained culture may be used as it is without separating the medium and the cells, or the cells may be recovered by separating the medium and the cells. The method for recovering the bacterial cells is not particularly limited. For example, if it is a liquid medium, it can be centrifuged, filtered, etc. If it is a solid medium, the bacterial cells can be recovered using a cell scraper or the like. Can do.

  The obtained bacterial cells of Lactobacillus plantarum may use live cells as they are, but it is preferable to use dead cells from the viewpoint of safety. The method for making dead cells is not particularly limited, and the culture containing the cells or the collected cells may be performed by a known method such as heat treatment, heat and pressure treatment, or ultraviolet treatment. Moreover, the obtained microbial cells may be further crushed or the crushed material may be further fractionated.

  Since the Lactobacillus plantarum obtained as described above has an excellent Endo180 production promoting action, it can be used as an active ingredient of an Endo180 production promoter.

[Endo180 production promoter]
The Endo180 production promoter of the present embodiment has one or more selected from the group consisting of the lotus germ extract, moon peach leaf extract, and Lactobacillus plantarum obtained as described above as active ingredients. Is. The Endo180 production promoter of this embodiment can be used for a wide range of uses such as pharmaceuticals, quasi drugs, foods and drinks, and cosmetics.

  The Endo180 production promoter of this embodiment may consist of a lotus germ extract, moon peach leaf extract or Lactobacillus plantarum or a mixture thereof, or a lotus germ extract, moon peach leaf extract or Lactobacillus plantarum. Alternatively, a mixture of these may be formulated.

  The Endo180 production promoter of this embodiment is an arbitrary pharmaceutically acceptable carrier such as dextrin, cyclodextrin, etc., and any other auxiliary agent according to a conventional method, such as powder, granule, tablet, liquid, etc. 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 promoter can be used by blending with other compositions (for example, foods and drinks, skin cosmetics, etc.), and can also be used as an ointment, a liquid for external use, a patch and the like.

  When the Endo180 production promoter of this embodiment is formulated, the content of the lotus germ extract, moon peach leaf extract or Lactobacillus plantarum or a mixture thereof is not particularly limited, and is appropriately determined according to the purpose. Can be set.

  In addition, the Endo180 production promoter of this embodiment may contain other natural extracts having an Endo180 production promoting action, together with a lotus germ extract, moon peach leaf extract, Lactobacillus plantarum, or a mixture thereof, as necessary. It can mix | blend and can be used as an active ingredient.

  Examples of the administration method of the Endo180 production promoter of this embodiment to a patient include transdermal administration, oral administration, and the like, but a suitable method for the prevention / treatment or the like may be appropriately selected according to the type of disease. . Further, the dose of the Endo180 production promoter 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.

  The Endo180 production promoter of this embodiment promotes the production of Endo180 in fibroblasts through the Endo180 production promotion action of the Lotus germ extract, moon peach leaf extract and Lactobacillus plantarum, which are active ingredients, By promoting the uptake of fragmented collagen by endocytosis to promote the regeneration of normal collagen fiber structure, it is possible to prevent, treat or ameliorate skin photoaging symptoms such as wrinkle formation. Further, the Endo180 production promoter of the present embodiment promotes regeneration of normal collagen fiber structure, thereby preventing arthritis such as rheumatoid arthritis, osteoarthritis, purulent arthritis, gouty arthritis, traumatic arthritis, osteoarthritis and the like. It can be used for applications such as prevention, treatment or improvement. However, the Endo180 production promoter of this embodiment can be used for all the uses meaningful in exhibiting the said effect | action besides these uses.

  Moreover, since the Endo180 production promoter of this embodiment has the outstanding Endo180 production promotion effect | action, it is suitable for mix | blending with food-drinks and a skin external preparation, for example. In this case, the lotus germ extract, moon peach leaf extract or Lactobacillus plantarum or a mixture thereof may be blended as it is, or the lotus germ extract, moon peach leaf extract or Lactobacillus plantarum or a mixture thereof. You may mix | blend the formulated Endo180 production promoter.

  Here, food and drink are those that are less likely to harm human health and are taken by oral or gastrointestinal administration in normal social life. It is not limited to such categories. Therefore, the “food and drink” in the present embodiment includes general foods taken orally, health foods (functional foods and drinks), health functional foods (specific health foods, nutritional functional foods, functional foods), pharmaceuticals It includes a wide range of quasi-drugs and pharmaceuticals.

  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. Specifically, for example, ointments, creams, and emulsions. , Lotions, cosmetic liquids, lotions, gels, cosmetic oils, packs, foundations, lip balms, bath salts, hair nicks, hair lotions, shampoos, rinses, soaps, body shampoos and the like.

  Moreover, since the Endo180 production promoter of this embodiment has the outstanding Endo180 production promotion effect | action, it can be utilized suitably also as a reagent for the research regarding these action mechanisms.

  The Endo180 production promoter of this embodiment is suitably applied to humans, but as long as each effect is exhibited, animals other than humans (for example, mice, rats, hamsters, dogs). , Cats, cattle, 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, lotus germ extract (manufactured by Maruzen Pharmaceutical, sample 1), moon peach leaf extract (manufactured by Maruzen Pharmaceutical, sample 2), and dead cells of Lactobacillus plantarum (maruzen) Samples 3) manufactured by Pharmaceutical Co., Ltd. were used (both lyophilized products).

[Test Example 1] Endo180 production promoting action test The above-mentioned natural product-derived components (Samples 1 to 3) were tested for Endo180 production promoting action as follows.

Human normal skin fibroblasts (NB1RGB) were cultured using Dulbecco's MEM containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with Dulbecco's MEM containing 0.25% FBS to a cell density of 1.0 × 10 ⁵ cells / mL, then seeded at 100 μL per well in a 96-well microplate, and cultured overnight.

  After completion of the culture, the medium was removed, and 100 μL of each test sample dissolved in Dulbecco's MEM containing 0.25% FBS (samples 1 to 3; see Table 1 for sample concentration) was added to each well and cultured for 24 hours. . As a control, culturing was similarly performed using Dulbecco's MEM containing 0.25% FBS to which no sample was added. After completion of the culture, the amount of Endo180 present on the cell surface of each well was measured by ELISA. From the measurement results, Endo180 production promotion rate (%) was calculated by the following formula.

Endo180 production promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Endo180 production amount with addition of test sample B: Endo180 production amount without addition of sample Table 1 shows the results.

  As shown in Table 1, the lotus germ extract (sample 1), moon peach leaf extract (sample 2), and Lactobacillus plantarum dead cells (sample 3) all have an excellent Endo180 production promoting action. Was confirmed.

  The Endo180 production promoter of the present invention can greatly contribute to the prevention, treatment or improvement of skin photoaging symptoms such as wrinkle formation; prevention, treatment or improvement of various arthritis.

Claims (1)

  An Endo180 production promoter comprising one or more selected from the group consisting of a lotus germ extract, a moon peach leaf extract, and a Lactobacillus plantarum as an active ingredient.