JP2008007412A - Involucrin production promoter and epidermal keratinization-normalizing agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an involucrin production promoter and an epidermal keratinization-normalizing agent by finding an ingredient having production promoting action of involucrin which is a CE (cornified envelope) precursor protein in epidermis from natural products and including the ingredient as an active ingredient. <P>SOLUTION: The present invention utilizes that one or more kinds of extracts of plants selected from yeast, Prunus persica B., Aloe ferox;africana Miller, Rosa multiflora Thunberg, Calendula officinalis L., Arnica montana L., Citrus aurantium L., Lavandula vera D.C., Hypericum erectum Thunb., or Hypericum perforatum L., Citrus unshiu Markovich, Melissa officinalis L., Luffa cylindrica M. Roemen and Equisetum arvense L. have involucrin production promoting action and karatinization-normalizing action. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

The present invention relates to an involucrin production promoter and an epidermal keratinization normalizing agent comprising an extract from a plant as an active ingredient.

  The skin has a function of alleviating external stimuli such as dryness, ultraviolet rays, and other physical and chemical stimuli and protecting the loss of body components such as moisture. This important function is called a skin barrier function, and the stratum corneum plays an important role in maintaining the function.

  The stratum corneum is formed from keratinocytes formed by terminal differentiation of epidermal keratinocytes and intercellular lipids that fill the space between the cells. Intercellular lipids mainly composed of ceramide have a stratum corneum barrier function by forming a lamellar structure. On the other hand, keratinocytes are mainly composed of keratin fibers and covered with a hydrophobic and strong cell membrane-like structure called a cornified envelope (keratin thickened membrane, hereinafter abbreviated as “CE”) which is a membrane lining protein. CE is formed by cross-linking and insolubilizing a plurality of CE precursor proteins such as involucrin and loricrin produced in the cells according to the differentiation of epidermal keratinocytes, and this CE is closely related to the barrier function of the skin. Is involved. Furthermore, ceramide and the like are covalently bonded to a part of them, and the base of the lamellar structure of intercellular lipids is supplied by taking a hydrophobic structure, thereby forming the basis of the stratum corneum barrier function and the skin water retention function .

  The keratinocytes are newly created one after another by the differentiation (keratinization) of the epidermal keratinocytes, and the old keratinocytes fall off from the stratum corneum as plaque. This is a phenomenon called skin turnover. It is said that the turnover cycle is about 4 weeks for normal skin, but if the turnover speed becomes abnormal due to the effects of aging, drying, ultraviolet rays, etc., the lamella structure is disturbed and CE is incomplete. In other words, the so-called aberrant keratinization is induced, the structure of corneocytes and intercellular lipids is abnormal, and the water retention function and barrier function of the stratum corneum are reduced. This is thought to lead to skin aging symptoms such as rough skin and dry skin. In patients with psoriasis and atopic dermatitis, immature CE is frequently observed in the skin area where the barrier function is reduced, and the correct formation of CE is very important for the barrier function of the skin. (Non-Patent Document 1).

That is, by promoting the production of transglutaminase and involucrin in the epidermis, keratinization of keratinocytes is promoted, and the formation of a healthy stratum corneum is promoted, so that the skin barrier function and water retention function are enhanced, dry skin, rough skin It is considered that skin symptoms such as atopic dermatitis and psoriasis can be prevented or ameliorated. Based on this idea, Ceylon tetsuboku seed extract (Patent Document 1), Kirinsai extract (Non-Patent Document 2) and the like are known as having an involucrin production promoting action.
JP-A-2005-213187 Experimental Dermatology 12: 591-601 (2003) Japanese Cosmetic Science Society Vol. 27: 6-10 (2003)

  However, the components that have been used to lower the stratum corneum barrier function so far have been effective in improving the skin barrier function by supplementing the components of intercellular lipids and forming a film on the skin. The improvement of the CE structure of the cells may not be sufficient, and it has been desired to develop a component that works to form a healthy CE.

  Therefore, in the present invention, an ingredient having an action of promoting the production of involucrin which is a CE precursor protein in the epidermis is found from natural products, and an involucrin production promoter and an epidermal keratinizing normalizing agent containing these as active ingredients are found. The purpose was to provide.

  In order to solve the above-mentioned problems, various studies have been carried out. The above extract has been found to promote involucrin production, and the present invention has been completed.

  According to the present invention, the composition contains one or more extracts selected from yeast, peach, aloe, Neubara, ginseng, arnica, daidai, lavender, hypericum, Satsuma mandarin, melissa, loofah and cedar as active ingredients. The involucrin production promoter has an excellent involucrin production promoting effect, and effectively prevents and improves skin symptoms such as dry skin, rough skin, atopic dermatitis, psoriasis due to normalization of epidermal keratinocytes. Can do.

  The involucrin production promoter of the present invention effectively uses one or more extracts selected from yeast, peach, aloe, Neubara, ginseng, arnica, daidai, lavender, hypericum, mandarin orange, melissa, loofah and horsetail. It is contained as a component, and further contains other components as necessary.

  The yeast is an autolysate of yeast (genus Saccharomyces).

  The peach is a plant of the Rosaceae family, and its scientific name is Prunus persica B. et al. It is. There is no restriction | limiting in particular in a structure part, According to the objective, it can select suitably, For example, a leaf, a stem, a flower, a bud, a seed, an above-ground part, whole grass, or a mixture thereof is mentioned. Among these, it is particularly preferable to use seeds.

  The aloe is a plant of the liliaceae family and has a scientific name of Aloe ferox; african miller. There is no restriction | limiting in particular in a structure part, According to the objective, it can select suitably, For example, a leaf, a stem, a flower, a bud, a seed, an above-ground part, whole grass, or a mixture thereof is mentioned. Among these, it is particularly preferable to use leaves.

  The rose is a plant of the Rosaceae family, and its scientific name is Rosa multiflora Thunberg. There is no restriction | limiting in particular in a structure part, According to the objective, it can select suitably, For example, a leaf, a stem, a flower, a cocoon, a seed, a fruit, a fruit skin, a fruit core, an above-ground part, whole grass, or a mixture thereof is mentioned. Among these, it is particularly preferable to use fruits.

  The milk moth is a plant of the Compositae family and has a scientific name of Calendula officinalis L. It is. There is no restriction | limiting in particular in a structure part, According to the objective, it can select suitably, For example, a leaf, a stem, a flower, a cocoon, a seed, a fruit, a fruit skin, a fruit core, an above-ground part, whole grass, or a mixture thereof is mentioned. Among these, it is particularly preferable to use flowers.

  Arnica is a plant of the Compositae family and has the scientific name Arnica montana L. It is. There is no restriction | limiting in particular in a structure part, According to the objective, it can select suitably, For example, a leaf, a stem, a flower, a cocoon, a seed, a fruit, a fruit skin, a fruit core, an above-ground part, whole grass, or a mixture thereof is mentioned. Among these, it is particularly preferable to use flowers.

  The Daidai is a plant of the Rutaceae family, and its scientific name is Citrus aurantium L. It is. There is no restriction | limiting in particular as a structure part, According to the objective, it can select suitably, For example, a leaf, a stem, a flower, a cocoon, a fruit, a fruit skin, a fruit nucleus, an above-ground part, whole grass, or a mixture thereof is mentioned. Among these, it is particularly preferable to use a peel.

  The lavender is a plant belonging to the family Labiatae, and the scientific name is Lavandula vera D. et al. C. It is. There is no restriction | limiting in particular in a structure part, According to the objective, it can select suitably, For example, a leaf, a stem, a flower, a root, a fruit, a fruit skin, a fruit nucleus, or a mixture thereof is mentioned. Among these, it is particularly preferable to use flowers.

  The hypericum is a plant of the family Glutiferae, and its scientific name is Hypericum electum Thumb. Or Hypericum performancea L. It is. There is no restriction | limiting in particular in a structural part, According to the objective, it can select suitably, For example, a leaf, a stem, a root, a flower, an above-ground part, whole grass, or a mixture thereof is mentioned. Among these, it is particularly preferable to use the above-ground part or whole grass.

  The Citrus unshiu is a plant belonging to the family Rutaceae, and its scientific name is Citrus unshiu Markovich. There is no restriction | limiting in particular in a structure part, According to the objective, it can select suitably, For example, a leaf, a stem, a branch, a flower, a cocoon, a fruit, a fruit skin, a fruit core, an above-ground part, whole grass, or a mixture thereof is mentioned. Among these, it is particularly preferable to use a peel.

  The Melissa is a plant of the Labiatae family and has a scientific name of Melissa officinalis L. It is. There is no restriction | limiting in particular in a structure part, According to the objective, it can select suitably, For example, a leaf, a stem, a root, a flower, a fruit, a fruit peel, a fruit nucleus, whole grass, or a mixture thereof is mentioned. Among these, it is particularly preferable to use leaves.

  The loofah is a plant of the Cucurbitaceae family, and its scientific name is Luffa cylindrica M. et al. Roemen. There are no particular restrictions on the constituent parts, and it can be appropriately selected according to the purpose, and examples thereof include leaves, stems, flowers, persimmons, fruits, pericarps, fruit nuclei, above-ground parts, or a mixture thereof. The aerial part is preferred.

  The horsetail is a plant belonging to the family Equisetaceae, and its scientific name is Equisetum arvense L. et al. It is. The constituent parts are not particularly limited and can be appropriately selected according to the purpose, but it is particularly preferable to use whole plants.

  Involucrin production containing as an active ingredient one or more extracts selected from the yeast, peach, aloe, Neubara, ginseng, Arnica, Daidai, Lavender, Hypericum, Unshimikan, Melissa, Loofah and Sugiina Although details of the accelerating agent are unknown, those active ingredients may be extracted by a method generally used for plant extraction, or commercially available products may be used. The extract includes any one of an extract obtained from an extraction raw material by an extraction process, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or a roughly purified product or a purified product thereof. Is also included.

  The extraction raw material is suitably dried and pulverized immediately after collection. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pre-processing, such as degreasing, with nonpolar solvents, such as hexane and benzene, you may use as an extraction raw material.

  As the solvent used for the extraction, it is preferable to use water, a hydrophilic organic solvent, or a mixture thereof at room temperature or a temperature not higher than the boiling point of the solvent. A component showing the involucrin production promoting action contained in each plant can be easily extracted by an extraction treatment using a polar solvent as an extraction 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. Therefore, the water that can be used as the extraction solvent in the present invention 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, propylene glycol, Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycerin, and a mixed solvent of the hydrophilic organic solvent and water can be used.

  In addition, when using the extraction solvent of the said water and a hydrophilic organic solvent, in the case of a lower alcohol, it is 1-90 mass parts with respect to 10 mass parts of water, and in the case of a lower aliphatic ketone, it is 10 mass parts of water. It is preferable to add 1-40 mass parts with respect to it. In the case of a polyhydric alcohol, it is preferable to add 1 to 90 parts by mass with respect to 10 parts by mass of water.

  In the present invention, it is not necessary to employ a special extraction method for extracting the involucrin production promoting substance from each of the extraction raw materials, and the extraction can be performed using an arbitrary apparatus at room temperature or under reflux heating.

  Specifically, each extraction raw material is put into a treatment tank filled with an extraction solvent, and further, while stirring occasionally as necessary, the mixture is allowed to stand for 30 minutes to 4 hours to elute soluble components, and then filtered and solidified. An extract is obtained by removing a thing, distilling an extraction solvent off from the obtained extract, and drying. The amount of the extraction solvent is usually 5 to 15 times (mass ratio) of the extraction raw material. The extraction condition is usually about 1 to 4 hours at 50 to 95 ° C. when water is used as the extraction solvent. Moreover, when using the mixed solvent of water and ethanol as an extraction solvent, it is about 30 minutes-4 hours at 40-80 degreeC normally. In addition, the extract obtained by extracting with a solvent is used as an active ingredient of the involucrin production promoter and / or epidermal keratinization normalizing agent of the present invention as it is if the extraction solvent is highly safe. However, it is easier to use a concentrated solution or a dried product thereof.

  The obtained extract is diluted, concentrated, dried, purified, etc. according to a conventional method in order to obtain a diluted or concentrated solution of the extract, a dried product of the extract, or a crude purified product or a purified product thereof. Processing may be performed.

  In addition, although the obtained extract can be used as it is as an involucrin production promoter and / or a skin keratinization normalizing agent, a concentrated solution or a dried product thereof is easier to use. In obtaining a dried product, a carrier such as dextrin or cyclodextrin may be added to improve hygroscopicity.

  In addition, since each extraction raw material has a unique odor and taste, it is possible to carry out purification for the purpose of decolorization, deodorization, etc. within a range that does not cause a decrease in its physiological activity, but it is added to cosmetics. In this case, since it is not used in large quantities, there is no practical problem even if it is not purified. Specifically, purification can be performed by activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, or the like.

  The involucrin production promoter and / or epidermal keratinization normalizing agent of the present invention can be made into any dosage form such as powder, granule, tablet, etc. by formulation.

  Examples of the present invention will be described below, but the present invention is not limited to these examples. In addition, the yeast extract, tonin extract, aloe extract, Agetsu extract-R, eucalyptus extract, arnica extract, spruce extract, lavender extract, hypericum extract-J used in this example. Each lyophilized product of Chimpi extract, Melissa extract-J, Loofah extract-JC and Sugina extract (all manufactured by Maruzen Pharmaceutical Co., Ltd.) was used as a sample.

[Test Example 1] Involucrin production promoting action test
Each of the above extracts was tested for involucrin production promoting action by the following method.
Normal human skin epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte medium (KGM) at 37 ° C. and 5% CO ₂ in an 80 cm ² flask, and the cells were collected by trypsin treatment. The collected cells are diluted with KGM to a cell density of 1.5 × 10 ⁴ cells / mL, then seeded at 200 μL per well in a 48-well plate, and cultured at 37 ° C. under 5% CO ₂ for 3 days. did.

After completion of the culture, the medium was removed, 200 μL of a sample solution dissolved in KGM was added to each well, and the cells were cultured at 37 ° C. under 5% CO ₂ for 48 hours. After completion of the culture, the medium was removed, the cells were fixed on a plate, and the amount of involucrin expressed on the cell surface was measured by ELISA using a monoclonal anti-human involucrin antibody. From the obtained measurement results, the involucrin production promotion rate (%) was calculated by the following formula.
Involucrin production promotion rate (%) = A / B × 100
In the formula, A represents “absorbance at a wavelength of 405 nm when a sample is added”, and B represents “absorbance at a wavelength of 405 nm when no sample is added”.

  The result of the said test is shown to Tables 1-13. In the above calculation formula, the involucrin production promotion rate without addition of the sample is 100%.

[Table 1]
Yeast extract added concentration Involucrin production promotion rate
100.0 μg / mL 119.6%
25.0 μg / mL 108.6%

[Table 2]
Increasing rate of involucrin production by adding peach seed extract
100.0 μg / mL 108.6%
25.0 μg / mL 106.2%

[Table 3]
Increasing rate of involculin production by adding aloe leaf extract
100.0 μg / mL 104.2%
25.0 μg / mL 106.3%

[Table 4]
Increased rate of involucrin production in fruit extract of rosehip
100.0 μg / mL 154.5%
25.0 μg / mL 135.9%

[Table 5]
Increasing rate of involucrin production with added concentration of flower extract
100.0 μg / mL 135.6%
25.0 μg / mL 120.9%

[Table 6]
Increasing rate of involcrine production with Arnica flower extract
25.0 μg / mL 136.2%

[Table 7]
Increased rate of involucrin production in Daidai peel extract
100.0 μg / mL 134.7%
25.0 μg / mL 144.1%

[Table 8]
Increasing rate of involucrin production by adding lavender flower extract
100.0 μg / mL 127.4%
25.0 μg / mL 100.9%

[Table 9]
Increasing rate of involcrine production by the concentration of hypericum whole plant extract
50.0 μg / mL 123.7%
12.5 μg / mL 109.7%

[Table 10]
Increasing rate of involucrin production in the concentration of Citrus unshiu peel extract
50.0 μg / mL 132.7%
12.5 μg / mL 141.2%

[Table 11]
Melissa leaf extract added concentration Involcrine production promotion rate
50.0 μg / mL 147.5%
12.5 μg / mL 117.6%

[Table 12]
Increased rate of involucrin production in the ground extract of loofah
12.5 μg / mL 112.3%

[Table 13]
Increasing rate of involucrin production by the concentration of whole plant extracts of horsetail
50.0 μg / mL 111.5%
12.5 μg / mL 115.8%

  As shown in Tables 1 to 13, the extract selected from yeast, peach, aloe, Neubara, ginseng, arnica, daidai, lavender, hypericum, Satsuma mandarin, Melissa, loofah and / or horsetail is high in invorcrine production. The promotion rate was shown. From these results, it was confirmed that these extracts have an excellent involucrin production promoting action.

  The involucrin production promoting agent and epidermal keratinization normalizing agent of the present invention normalizes epidermal keratinocytes by involucrin production promoting action in the epidermis, thereby reducing skin symptoms such as dry skin, rough skin, atopic dermatitis, and psoriasis. Can greatly contribute to prevention and improvement.

Claims (2)

It contains one or more extracts selected from yeast, peach, aloe, Neubara, ginseng, arnica, daidai, lavender, hypericum, unshimikan, melissa, loofah and horsetail as an active ingredient Involucrin production promoter. A skin keratinizing normalizing agent comprising the extract according to claim 1 as an active ingredient.