JP2018108973A - Type xvii collagen production promoter - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide type XVII collagen production promoters containing as an active ingredient a naturally occurring component having type XVII collagen production promoting action by finding out it among naturally occurring components having excellent safety.SOLUTION: The extract from Origanum majorana is incorporated as an active ingredient of type XVII collagen production promoter according to the present invention.SELECTED DRAWING: None

Description

  The present invention relates to a type XVII collagen production promoter.

  The skin is composed of the stratum corneum, epidermis, basement membrane and dermis. The basement membrane is present at the boundary between the epidermis and the dermis and plays an important role not only in connecting the epidermis and the dermis but also in maintaining the skin function (see Non-Patent Document 1). The main skeleton of the basement membrane has a network structure composed of type IV collagen. A structure called a hemidesmosome contributes to the adhesion between the basement membrane and the epidermal cells, and the basement membrane and the epidermal cells are adhered by cell adhesion molecules such as XVII collagen, integrin α6β4, laminin 5 and the like. In young skin, the basement membrane works to maintain the homeostatic interaction of the epidermis and dermis, ensuring moisture retention, flexibility, elasticity, and the like. Maintained.

  However, when there is an influence of certain external factors such as ultraviolet irradiation, drastic air drying, excessive skin washing, etc., or when aging progresses, the cell adhesion molecule degrades and deteriorates, and the basement membrane structure Is destroyed (see Non-Patent Documents 2 and 3). As a result, the skin's moisturizing function and elasticity are lowered, and the keratin begins to exfoliate abnormally, so the skin loses its tension and gloss, and exhibits aging symptoms such as roughness and wrinkles. In this way, changes accompanying skin aging, that is, formation of wrinkles, disappearance of texture, decrease in elasticity, etc. are associated with a decrease in basement membrane components and structural changes in the basement membrane. It is considered that skin aging symptoms can be prevented and improved by promoting production.

  In recent years, type XVII collagen expressed in hair follicle stem cells is essential for the maintenance of hair follicle stem cells, and hair follicle stem cells maintain pigment stem cells. When collagen XVII is degraded with age, hair follicle stem cells lose stem cell properties and differentiate into epidermal keratinocytes, causing hair follicles to shrink and causing hair loss (See Non-Patent Documents 4 and 5), and the relationship between type XVII collagen and white hair, thin hair, and the like has attracted attention. Under such circumstances, a hair loss inhibitor, a hair depigmentation inhibitor, and the like containing a type XVII collagen or a variant thereof have been proposed (see Patent Document 1).

JP 2009-161509 A

Marinkovich MP et al., J. Cell Biol., 1992, vol. 199, pp. 695-703 Lavker et al., J. Invest. Dermatol., 1979, vol. 73, pp. 59-66 Langton AK et al., Mech. Aging Dev., 2016, vol.156, pp.14-6 Tanimura S et al., Cell Stem Cell, 2011, vol.8, pp.177-87 Matsumura H et al., Science, 2016, vol.351, aad4395

  It is an object of the present invention to provide an XVII type collagen production promoter comprising an active ingredient as an active ingredient, which has been found to have an action of promoting the production of type XVII collagen among natural product-derived components having excellent safety. And

  In order to solve the above-mentioned problems, the type XVII collagen production promoter according to the present invention contains an extract from marjoram as an active ingredient.

  According to the present invention, by containing an extract from marjoram as an active ingredient, it is possible to provide an XVII type collagen production promoter that is excellent in action effect and high in safety.

Embodiments of the present invention will be described below.
The XVII type collagen expression promoting agent of this embodiment contains an extract from marjoram as an active ingredient.

  Here, in the present embodiment, the “extract” refers to an extract obtained using the above plant as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or these Either a crude product or a purified product is included.

  The extraction raw material used in this embodiment is marjoram (scientific name: Origanum majorana).

  Marjoram (Origanum majorana, also known as: Majorana, Oregano) is a perennial herb belonging to the family Lamiaceae, which is distributed in the Mediterranean coast, North Africa, West Asia, etc., and can be easily obtained from these regions it can. Examples of the constituent parts of marjoram that can be used as an extraction raw material include a leaf part, a stem part, a flower part, a bud part, a rhizome part, an above-ground part, or a mixture thereof, and preferably a leaf part.

  The extract from the marjoram 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 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.

  The extract from the plant can be obtained by drying the plant as an 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.

  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 5: 5 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.

  The extract from marjoram obtained as described above (hereinafter sometimes simply referred to as “marjoram extract”) has an excellent XVII collagen production promoting action, and therefore, an XVII collagen production promoter. It can be used as an active ingredient. The XVII type collagen production promoter according to this embodiment can be used for a wide range of uses such as pharmaceuticals, quasi drugs, and cosmetics.

  The XVII type collagen production promoter according to the present embodiment may be composed solely of marjoram extract or may be formulated from marjoram extract.

  XVII type collagen production promoter according to the present embodiment, using a pharmaceutically acceptable carrier such as dextrin, cyclodextrin and other optional auxiliaries, according to conventional methods, powder, granule, tablet, liquid, etc. Can be formulated into any 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 type XVII collagen production promoter can be used by blending with other compositions (for example, skin cosmetics, etc.), and can also be used as tablets, capsules and the like.

  When the XVII collagen production promoter according to the present embodiment is formulated, the content of the marjoram extract is not particularly limited and can be appropriately set according to the purpose.

  In addition, the XVII type collagen production promoter according to the present embodiment can be used as an active ingredient by blending other natural extracts having an XVII type collagen production promoting action together with the marjoram extract, if necessary. .

  Examples of the administration method to the patient of the XVII type collagen production promoter according to this embodiment include oral administration and intravenous administration, etc., but a suitable method for the prevention or treatment is appropriately selected depending on the type of disease. do it. In addition, the dose of the XVII collagen production promoter according to 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 XVII type collagen production promoter according to this embodiment can promote the production of type XVII collagen through the XVII type collagen production promoting action of the marjoram extract, which is an active ingredient. This induces remodeling of the basement membrane structure, and can prevent or improve skin aging symptoms such as wrinkle formation, loss of texture, reduced elasticity, and treat or improve skin damage Can do. In addition, the type XVII collagen production promoter according to the present embodiment can maintain hair follicle stem cells and pigment stem cells through the XVII type collagen production promoting action of the marjoram extract, thereby depigmenting hair (white hair). And can prevent or improve hair loss. However, the XVII type collagen production promoter according to the present embodiment can be used for all purposes other than these uses, which are meaningful for exerting the XVII type collagen production promoting action.

  Since the XVII type collagen production promoter of this embodiment has the outstanding XVII type collagen production promotion effect | action, it is suitable for mix | blending with a skin external preparation or food-drinks, for example. In this case, the marjoram extract may be blended as it is, or an XVII type collagen production promoter formulated from the marjoram extract may be blended.

  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, lotions, serums, lotions, gels, beauty oils, packs, foundations, lip balms, bath salts, hair nicks, hair lotions, soaps, body shampoos and the like.

  The food and drink are not limited in its category, and include a wide range of foods that are taken orally, such as general foods, health foods (functional foods and drinks), health functional foods (food for specified health use, nutritional functional foods) and the like.

  Moreover, since the XVII type collagen production promoter which concerns on this embodiment has the outstanding XVII type collagen production promotion effect | action, it can be utilized suitably also as a reagent for the research regarding these action mechanisms.

  The XVII collagen production promoter according to the present embodiment is preferably applied to humans, but as long as the respective effects are exhibited, animals other than humans (for example, mice, rats, 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 this test example, a lyophilized product (sample 1) of marjoram extract (manufactured by Maruzen Pharmaceutical Co., Ltd.) was used as a test sample.

[Test Example 1] Type XVII collagen (COL17A1) mRNA expression promoting effect test The marjoram extract (sample 1) was tested for COL17A1 mRNA expression promoting action as follows.

Normal human neonatal foreskin keratinocytes (NHEK) were cultured in a 75 cm ² flask using normal human epidermal keratinocyte growth medium (Humdia-KG2, hereinafter referred to as “KGM”) at 37 ° C. and 5% CO ₂ . The cells were precultured under conditions, and the cells were collected by trypsin treatment. The collected cells are diluted with KGM so as to have a cell density of 15 × 10 ⁴ cells / mL, then seeded in 2 mL each in a 35 mm petri dish (30 × 10 ⁴ cells / 2 mL / petri dish), 37 ° C., 5% CO _2. Cultured overnight under the conditions of After culturing, the medium is replaced with a basal medium for normal human epidermal keratinocyte proliferation (Humdia-KB2, hereinafter referred to as “KBM”, which does not contain growth factors (hEGF, BPE, insulin) added to KGM), The culture was further continued for 24 hours.

After culturing for 24 hours, the test sample (sample 1, sample concentration: see Table 1 below) was replaced with dissolved KBM, and cultured under conditions of 37 ° C. and 5% CO ₂ for 24 hours. In addition, it culture | cultivated similarly using KBM without a sample as control. After culturing, the medium is removed, and total RNA is extracted with ISOGEN II (Nippon Gene, Cat. No. 311-07361), and the amount of each RNA is measured with a spectrophotometer so that it becomes 200 ng / μL. Total RNA was prepared.

  Using this total RNA as a template, mRNA expression levels were measured for COL17A1 and GAPDH as an internal standard. Detection was performed by real-time 2 Step RT-PCR reaction using TaKaRa SYBR Prime Script RT-PCR kit (Perfect Real Time) (Takara Bio, code No. RR063A) using a real-time PCR device Smart Cycler (Cepheid). . The expression level of COL17A1 mRNA was determined as a GAPDH value based on total RNA preparations prepared from cells cultured in “addition of test sample” and “no sample addition”, respectively. From the obtained value, the COL17A1 mRNA expression promotion rate (%) was calculated by the following formula.

COL17A1 mRNA expression promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Correction value with addition of test sample B: Correction value without addition of sample Table 1 shows the results.

  As shown in Table 1, it was confirmed that the marjoram extract (sample 1) has an excellent COL17A1 mRNA expression promoting action.

[Test Example 2] XVII type collagen production promoting action test The marjoram extract (sample 1) was tested for XVII type collagen production promoting action as follows.

Normal human neonatal foreskin keratinocytes (NHEK) were cultured using KGM, and then cells were collected by trypsin treatment. The collected cells were diluted with KBM to a cell density of 2 × 10 ⁵ cells / mL, then seeded at 100 μL per well in a 96-well plate, and cultured overnight.

  After completion of the culture, the medium was removed, and 100 μL of a test sample dissolved in KBM (sample 1, refer to Table 2 below for sample concentration) was added to each well and cultured for 48 hours. In addition, it culture | cultivated similarly using KBM without a sample as control. After completion of the culture, the medium is removed, the cells are fixed to the plate, and the amount of XVII collagen expressed on the cell surface is measured by ELISA using an anti-human XVII collagen α1 chain polyclonal antibody (manufactured by FLAREBIO BIOTECH). did. From the measurement results, the type XVII collagen production promotion rate (%) was calculated by the following formula.

Type XVII collagen production promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Type XVII collagen amount with addition of test sample B: Type XVII collagen amount without addition of sample Table 2 shows the results.

  As shown in Table 2, it was confirmed that the marjoram extract (sample 1) has an excellent XVII collagen production promoting action.

  The type XVII collagen production promoter according to the present invention can greatly contribute to reconstruction of the basement membrane structure; prevention or improvement of skin aging symptoms; treatment or improvement of skin damage; prevention of white hair; prevention of hair loss;

Claims (1)

  A type XVII collagen production promoter characterized by containing an extract from marjoram as an active ingredient.