JP2008007411A - Transglutaminase production promoter and epidermal keratinization-normalizing agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a transglutaminase production promoter and an epidermal keratinization-normalizing agent by finding an ingredient having enzyme transglutaminase production promoting action essential for CE (cornified envelope) formation 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 Equisetum arvense L., Luffa cylindrica M. Roemen, Citrus aurantium L., Paeonia suffruticosa Andrews, Citrus unshiu Markovich, Hypericum erectum Thunb. or Hypericum perforatum L, Morus bombycis Koidzumi, Curcuma longa L., Melissa officinalis L., Lavandula vera D.C., Ganoderma lucidum(Fr.)Karst. and Rosmarinus officinalis L. have transglutaminase production promoting action and karatinization-normalizing action. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

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

The skin has a function of mitigating 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 some of them, and the base of the lamellar structure of the intercellular lipid is supplied by taking a hydrophobic structure, and the basis of the stratum corneum barrier function and the moisture retention function of the skin is formed. .

  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, as one having a transglutaminase production-promoting action, Hunan cocoon tea extract and its acid hydrolyzate (patent document 1), phlorizin and phloretin (patent document 2), vitamin D and related compounds (patent) Document 3), extracts of genus Oenothera (Patent Document 4) and the like are known.
Japanese Patent Application No. 2005-292445 Japanese Patent Application No. 2005-300388 JP 2004-115451 A JP 2004-91376 A Experimental Dermatology 12: 591-601 (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, a component having an action of promoting the production of enzyme transglutaminase essential for CE formation in the epidermis is found from natural products, and a transglutaminase production promoter containing this as an active ingredient and normalization of epidermal keratinization The purpose was to provide an agent.

  In order to solve the above-mentioned problems, various studies have been conducted. As a result, one or more kinds selected from Sugina, Loofah, Daidai, Button, Satsumikan, Hypericum, Mulberry, Turmeric, Melissa, Lavender, Reishi and Rosemary are selected. A plant extract was found to promote transglutaminase production, and the present invention was completed.

  According to the present invention, transglutaminase containing one or more plant extracts selected from cedar, loofah, daidai, button, shrimp, hypericum, mulberry, turmeric, melissa, lavender, litchi and rosemary. The production promoter has an excellent epidermal transglutaminase production promoting effect, and can effectively prevent and improve skin symptoms such as dry skin, rough skin, atopic dermatitis, psoriasis due to normalization of epidermal keratinocytes it can.

  The transglutaminase production promoter of the present invention contains one or more plant extracts selected from cedar, loofah, daidai, button, mandarin orange, hypericum, mulberry, turmeric, melissa, lavender, litchi and rosemary. In addition, it contains other components as required.

  The horsetail is a plant belonging to the family Equisetaceae, and its scientific name is Equisetum arvense L. et al. It is. There is no restriction | limiting in particular in a structure part, According to the objective, it can select suitably, Especially the whole plant is preferable.

  The loofah is a plant of the Cucurbitaceae family, and its scientific name is Luffa cylindrica M. et al. Roemen. 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 fruit, a fruit skin, a fruit core, an above-ground part, or these mixtures are mentioned. Among these, it is particularly preferable to use the above-ground part.

  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 in a structure part, According to the objective, it can select suitably, For example, a leaf, a stem, a flower, a bud, a fruit, a fruit skin, a fruit core, an above-ground part, whole grass, or these mixtures are mentioned. Of these, the peel is particularly preferred.

  The button is a plant of the Paeoniaceae family and the scientific name is Paeonia suffrificosa Andrews. 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 root bark, an above-ground part, or these mixtures are mentioned. Of these, root bark is particularly preferred.

  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 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 preferable to use the above-ground part or whole grass.

  The mulberry is a plant belonging to the family Moraceae, and its scientific name is Morus bombycis Koidzumi. 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 root bark, an above-ground part, or these mixtures are mentioned. Of these, root bark is particularly preferred.

  The turmeric is a plant belonging to the family Zingiberaceae and has a scientific name of Curcuma longa 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 bud, a rhizome, an above-ground part, or these mixtures are mentioned. Among these, it is particularly preferable to use a rhizome.

  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 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 litchi is a plant of the Polypoaceae family, and has a scientific name of Ganoderma lucidum (Fr.) Karst. It is. There is no restriction | limiting in particular in a structure part, According to the objective, it can select suitably, It is preferable to use a fruit body especially.

  The rosemary is a plant of the Labiatae family and has a scientific name of Rosmarinus 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 bud, a fruit, a fruit skin, a fruit core, an above-ground part, whole grass, or these mixtures are mentioned. Among these, it is particularly preferable to use leaves.

  In addition, transglutaminase which is an active ingredient contained in one or more plant extracts selected from the above cedar, loofah, daidai, button, unshimikan, hypericum, mulberry, turmeric, melissa, lavender, litchi and rosemary Although details of the production promoting substance 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 transglutaminase 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 transglutaminase production promoting substance from each extraction raw material, 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, if the extraction solvent is highly safe, is blended as it is as an active ingredient of the transglutaminase production promoter and / or epidermal keratinization normalizing agent of the present invention. Although it can be used, 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 a transglutaminase 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 transglutaminase 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 said horsetail extract, loofah extract-JC, spruce extract, button pi extract, chimpi extract, hypericum extract-J, Soraku extract, turmeric extract-J, Melissa extract used in this Example Each lyophilized product of liquid-J, lavender extract, litchi extract and rosemary extract (all manufactured by Maruzen Pharmaceutical Co., Ltd.) was used as a sample.

[Test Example 1] Transglutaminase production promoting action test
Each of the above extracts was tested for the effect of promoting transglutaminase production 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 1.0 × 10 ⁵ cells / mL, then seeded at 100 μL per well in a 96-well plate, and cultured at 37 ° C. under 5% CO ₂ for 2 days. did.

After completion of the culture, 100 μL of a sample solution dissolved in KGM was added to each well and cultured at 37 ° C. under 5% CO ₂ for 24 hours. After completion of the culture, the medium was removed, the cells were fixed on a plate, and the amount of transglutaminase expressed on the cell surface was measured by ELISA using a monoclonal anti-human transglutaminase-1 antibody. From the obtained measurement results, the transglutaminase production promotion rate (%) was calculated by the following formula.
Transglutaminase 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 1-12. In the above calculation formula, the rate of acceleration of transglutaminase production without addition of the sample is 100%.

[Table 1]
Transglutaminase production promotion rate of added concentration of Japanese horsetail extracts
100.0 μg / mL 106.1%
25.0 μg / mL 114.1%

[Table 2]
Transglutaminase production promotion rate of added concentration of aboveground extract of loofah
50.0 μg / mL 104.8%
12.5 μg / mL 109.3%

[Table 3]
Daigluta pericarp extract concentration concentration transglutaminase production promotion rate
50.0 μg / mL 114.8%
12.5 μg / mL 114.3%

[Table 4]
Button root bark extract addition concentration transglutaminase production promotion rate
50.0 μg / mL 101.9%
12.5 μg / mL 117.0%

[Table 5]
Transglutaminase production promotion rate of added concentration of Citrus unshiu peel extract
50.0 μg / mL 116.8%
12.5 μg / mL 104.6%

[Table 6]
Transglutaminase production promotion rate of added concentration of hypericum whole grass extract
50.0 μg / mL 112.4%
12.5 μg / mL 110.7%

[Table 7]
Mulberry root bark extract added concentration transglutaminase production promotion rate
50.0 μg / mL 107.1%
12.5 μg / mL 107.4%

[Table 8]
Accelerated rate of transglutaminase production in turmeric rhizome extract
50.0 μg / mL 109.6%
12.5 μg / mL 118.9%

[Table 9]
Melissa leaf extract added concentration transglutaminase production promotion rate
50.0 μg / mL 104.0%
12.5 μg / mL 112.2%

[Table 10]
Lavender flower extract added concentration transglutaminase production promotion rate
50.0 μg / mL 113.3%
12.5 μg / mL 107.2%

[Table 11]
Transglutaminase production promotion rate of litchi fruiting body extract
50.0 μg / mL 112.2%
12.5 μg / mL 110.6%

[Table 12]
Increased transglutaminase production rate of rosemary leaf extract
50.0 μg / mL 105.2%
12.5 μg / mL 114.8%

  As shown in Tables 1 to 12, one or two or more plant extracts selected from horsetail, loofah, daidai, button, mandarin orange, hypericum, mulberry, turmeric, melissa, lavender, lychee and rosemary, Also showed a high transglutaminase production promotion rate. From these results, it was confirmed that these extracts have an excellent transglutaminase production promoting action.

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

Claims (2)

Trans, comprising as an active ingredient one or more plant extracts selected from Sugina, Loofah, Daidai, Button, Satsuma mandarin, Hypericum, Mulberry, Turmeric, Melissa, Lavender, Ganoderma and Rosemary Glutaminase production promoter. A skin keratinizing normalizing agent comprising the plant extract according to claim 1 as an active ingredient.