JP2021176881A - Filaggrin production promoter - Google Patents

Abstract

To provide a composition that promotes production of filaggrin, promotes skin keratinization, maintains skin moisture, suppresses wrinkle formation, prevents rough skin, normalizes skin texture, and improves skin problems such as atopic dermatitis.SOLUTION: A filaggrin production promoter comprises at least one kind of an extract of Isodon japonicus and an extract of Fucus sp. as an active ingredient.SELECTED DRAWING: Figure 1

Description

The present invention promotes the production of filaggrin, involucrin, loricrin, corneodesmosin, promotes skin keratinization, maintains skin moisture, suppresses wrinkle formation, prevents rough skin, normalizes skin texture, atopic dermatitis, etc. Concers on compositions that improve skin problems.

The skin is located on the outermost side of the body and acts as a barrier against external stimuli such as bacteria. In the skin, epidermal cells keratinize from basal cells to spiny cells, granule cells, and even stratum corneum cells over a period of about 4 weeks, and in these cells intercellular lipids, NMFs (natural moisturizing factors), and even The barrier function is achieved by forming a cornified envelope.
Incomplete response of the stratum corneum to the aging of the skin and changes in the external environment causes the formation of wrinkles, abnormal texture, rough skin, etc., but the decrease in the water content of the stratum corneum is an important factor.
Natural moisturizing factors are involved in the water retention of keratin, and most of these natural moisturizing factors are related to amino acids produced by protein degradation.
Amino acids, which are the main components of this natural moisturizing ingredient, are produced by the decomposition of filaggrin derived from keratohyalin granules in the stratum corneum. This filaggrin is expressed as profilaggrin in epidermal keratinocytes, is immediately phosphorylated, and accumulates in keratohyalin granules. After that, it is decomposed into filaggrin through dephosphoric acid and hydrolysis, and migrates to the stratum corneum, increases the aggregation efficiency of keratin filaments, and is involved in the internal construction of corneocytes. In recent years, it has been clarified that this filaggrin is very important and indispensable for water retention of the skin, and that the synthetic ability of filaggrin decreases due to conditions such as dryness, and the amount of amino acids in the stratum corneum decreases.
When profilaggrin, which constitutes keratohyalin granules existing in the stratum granulosum of the epidermis, is keratinized during the process of keratinization of epidermal cells, it is decomposed by the action of dephosphorylation and protease to release a protein called filaggrin, and the arginine residue of the filaggrin. Is modified by an enzyme called peptidylarginine deiminase to become a citrulin residue, and filaggrin is gradually separated from the keratin fibers and decomposed to produce an NMF component.
In addition, trans-urocanic acid is produced from histidine produced by the decomposition of filaggrin by the action of histidase, which protects the skin from ultraviolet rays.
Thus, the promotion of filaggrin synthesis plays an important role in the water retention of the stratum corneum and the protective function from ultraviolet rays.

In addition, the epidermis has a protective function of protecting the skin from various stimuli from the outside world by constantly producing new keratinocytes by division of keratinocytes and subsequent differentiation. In particular, in the process of differentiation of keratinocytes, proteins such as involucrin are expressed from the spinous layer to the granular layer to form a cornified envelope (CE), which is an insoluble cell membrane-like structure that encloses the keratinocytes, and the keratinocytes. Contributes to the stability of the cell skeleton and structure of.
However, when the amount of involucrin produced in the epidermis decreases due to various factors, the formation of the cornified envelope (CE) becomes incomplete and keratinization does not occur normally. As a result, it is considered that the keratin barrier function and the moisturizing function of the skin are deteriorated, and skin symptoms such as rough skin and dry skin are exhibited.
Therefore, by increasing the production of involucrin in the epidermis of keratinocytes, promoting the formation of cornified envelopes and normalizing keratinization, the skin barrier function associated with external stimuli such as desiccation and ultraviolet rays It is thought that various skin symptoms such as dry skin and rough skin can be prevented and ameliorated by suppressing the decrease in skin.

Loricrin is a major component of the cornified cell envelope, and its expression is one of the important constituent proteins of the peripheral zone, starting from the keratohyalin granules and gradually being incorporated into the peripheral zone, and the formation of natural moisturizing factors. It is involved in various stratum corneum functions including the above, and suppresses the formation of wrinkles, abnormal texture, and rough skin.

Desmosomes play a role in connecting cells, but in the stratum corneum of the skin, Corneodesmosomes, which are slightly modified from normal desmosomes, play a role. Corneodesmosin is a protein located in the center of Corneodesmosomes and is known to bind to the cornified envelope. Enzymatic degradation of corneodesmosomes leads to exfoliation of the stratum corneum in the uppermost layers of the skin.
Therefore, the decrease in the expression of the desmosome-constituting protein accompanying skin aging causes wrinkle formation, the decrease in the texture pattern, and the rough skin. Therefore, it is necessary to promote the expression of the desmosome-constituting protein such as corneodesmosin.
By promoting the production of filaggrin, involucrin, loricrin, and corneodesmosin in this way, an external skin preparation effective for normalizing skin function, suppressing wrinkle formation, preventing rough skin, normalizing texture, atopic dermatitis, etc. can be obtained. Be done.
Although some substances that promote these productions are known, none have been sufficiently satisfied.

Sagarame (Eisenia arborea Areschoug) is a seaweed of brown algae, kelp family (Laminariaceae), and genus Arame (Eisenia). It is used as an ingredient.
Further, applications such as an angiogenesis inhibitor, β-glucuronidase inhibitor, AGE production inhibitor, and aquaporin production-enhancing preparation are known. (See Patent Documents 1 to 4)

Hibamata is a seaweed that belongs to the family Fucales of the order Brown algae, and is a seaweed that has been used for food since ancient times in Europe and is rich in minerals. Among them, it has a high content of iodine and zinc, and is said to be useful for supplementing these components. In addition, mucilage is a raw material for alginate esters required in the manufacture of foods, cosmetics and pharmaceuticals. Further, it is known to have actions such as hyaluronidase inhibition, collagen gel contraction promotion, integrin expression promotion, and elastase inhibition. (Patent Documents 5 to 8)
In Europe, Fucus vesiculosus is mainly used and is also called Seaweed. Fucus evanescens is distributed in Japan.

Black tea is made from tea (Camellia sinensis) leaves and fermented picked tea leaves.
Of course, it is mainly used for drinking, but applications such as a plaque formation inhibitor, a melanin production inhibitor, an allergen inactivating agent, and an OPH activity enhancer are also known. (See Patent Documents 9 to 12)

Enmeisou (Lamiaceae) is a Japanese folk medicine that dried the whole plant of Isodon japonicus of the Labiatae family and has been used for indigestion, loss of appetite, abdominal pain, etc. Further, applications such as anti-allergic agents, antihistamines, anti-complement activators, anti-obesity cosmetics, platelet aggregation inhibitory action, angiogenesis inhibitor, tight junction formation promoter and the like are known. (Patent Documents 13 to 18)

Japanese Unexamined Patent Publication No. 2006-022033 Japanese Unexamined Patent Publication No. 2006-04518 Japanese Unexamined Patent Publication No. 2008-21246 Japanese Unexamined Patent Publication No. 2013-087057 Japanese Unexamined Patent Publication No. 09-06726 Japanese Unexamined Patent Publication No. 10-072336 JP-A-11-246428 Japanese Unexamined Patent Publication No. 2000-053578 Japanese Unexamined Patent Publication No. 2001-354545 Japanese Unexamined Patent Publication No. 08-301722 Japanese Unexamined Patent Publication No. 08-092057 Japanese Unexamined Patent Publication No. 05-262636 Japanese Unexamined Patent Publication No. 07-196522 Japanese Unexamined Patent Publication No. 09-087189 Japanese Unexamined Patent Publication No. 10-203990 Japanese Unexamined Patent Publication No. 2007-197388 Japanese Unexamined Patent Publication No. 2012-025777 Japanese Unexamined Patent Publication No. 2013-056841

An object of the present invention is an external preparation for skin that is effective for normalizing skin function, suppressing wrinkle formation, preventing rough skin, normalizing skin texture, atopic dermatitis, etc. by promoting the production of filaggrin, involucrin, loricrin, and corneodesmosin. Is to get.

As a result of diligent studies by the present inventors, it was found that an extract of rockweed, an extract of rockweed, an extract of black tea, and an extract of Enmeisou achieve the above-mentioned object.
It is preferable that the rockweed, rockweed, black tea, and Enmeisou are extracted after being dried and then subjected to treatments such as shredding, drying, and crushing in consideration of extraction efficiency.
Drying may be performed in the sun or using a commonly used dryer.
As the solvent used for the extraction, water, a hydrophilic organic solvent, or a mixture thereof is used.
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 those subjected to various treatments. Treatments applied to water include, for example, 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 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 and 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.
When a mixed solvent of the water and the hydrophilic organic solvent is used, the amount of lower alcohol is 3 to 20 parts by mass with respect to 10 parts by mass of water, and the amount of lower aliphatic ketone is 10 parts by mass of water. On the other hand, it is preferable to add 3 to 15 parts by mass. In the case of polyhydric alcohol, it is preferable to add 3 to 20 parts by mass with respect to 10 parts by mass of water.
The amount of the organic solvent used for extraction is preferably about 5 to 100 times, more preferably about 10 to 50 times the amount of the plant as a raw material. In order to further increase the extraction efficiency, stirring or homogenization may be performed in the extraction solvent. It is appropriate that the extraction temperature is from about 5 ° C. to a temperature equal to or lower than the boiling point of the extraction solvent. The extraction time varies depending on the type of extraction solvent and the extraction temperature, but it is appropriate to set it to about 1 hour to 14 days.
The extraction operation is not limited to a one-time operation. A fresh solvent can be added again to the residue after extraction to perform an extraction operation, or the extraction solvent can be brought into contact with the extraction raw material multiple times.
If necessary, further purification treatment such as deodorization and decolorization may be added within a range that does not affect the effect, and the concentration can be achieved by a vacuum concentrator such as an evaporator or solvent removal by heating.
It is also possible to further purify this extract using a synthetic adsorbent (Diaion HP20, Sephadex SP825, Amberlite XAD4, MCIgelCHP20P, etc.), dextran resin (Sephadex LH-20, etc.), ultrafiltration, etc. be.

Although the pharmaceutical product of the present invention exhibits medicinal effects by oral, injectable, or external use, it is preferably used as a skin external preparation. External dermatological agents include skin cosmetics, quasi-drugs for external use, and external dermatological agents for medical use.

In addition to the above ingredients, the formulations of the present invention include surfactants, oily ingredients, moisturizers, polymer compounds, ultraviolet absorbers, anti-inflammatory agents, and bactericides used in various pharmaceutical and cosmetic formulations. Antioxidants, metal ion blockers, preservatives, vitamins, pigments, fragrances, water and the like can be blended.

As the above-mentioned surfactant, any of anionic, cationic, nonionic, natural and synthetic surfactants can be used, but it is preferable to use nonionic surfactants in consideration of skin irritation. Examples of the nonionic surfactant include glycerin fatty acid ester, propylene glycol fatty acid ester, sorbitan fatty acid ester, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene sorbit fatty acid ester, polyoxyethylene alkyl ether, and polyoxyethylene polyoxypropylene glycol. , Polyoxyethylene polyoxypropylene alkyl ether, polyethylene glycol fatty acid ester, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, alkyl glycoside and the like.

Examples of the oily component include fats and oils, waxes, hydrocarbons, higher fatty acids, higher alcohols, esters, essential oils, silicone oils and the like. Examples of fats and oils include soybean oil, nuka oil, jojoba oil, avocado oil, almond oil, olive oil, cacao oil, sesame oil, persic oil, castor oil, palm oil, minced oil, beef fat, pork fat and other natural fats and oils. Hardened oil obtained by hydrogenating natural fats and oils from Japan and synthetic triglycerides such as myristic acid glyceride and 2-ethylhexanoic acid triglyceride; Examples of waxes include carnauba wax, whale wax, honeydew, lanolin and the like; hydrocarbons Examples include liquid paraffin, vaseline, paraffin microcrystalin wax, selecin, squalane, bristan, etc .; higher fatty acids include, for example, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, linoleic acid, etc. Linolenic acid, lanolinic acid, isostearic acid and the like; higher alcohols include, for example, lauryl alcohol, cetyl alcohol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, 2-hexyldecanol and the like; esters include, for example, cetyl octanate. , Octanoic triglyceride, myristyl lactate, cetyl lactate, isopropyl myristate, myristyl myristate, octyldodecyl myristate, isopropyl palmitate, isopropyl adipate, butyl stearate, decyl oleate, cholesterol isostearate, POE sorbit fatty acid ester, etc. The essential oils include, for example, peppermint oil, jasmine oil, show brain oil, hinoki oil, peppermint oil, ryu oil, terepine oil, coconut skin oil, bergamot oil, orange oil, shove oil, pine oil, lavender oil, and bay oil. , Clove oil, hiba oil, rose oil, eucalyptus oil, lemon oil, thyme oil, peppermint oil, rose oil, sage oil, menthol, cineole, eugenol, citral, citroneral, borneol, linalol, geraniol, camphor, timol, spirantol, Pinen, limonene, terpene-based compounds and the like; Examples of silicone oils include dimethylpolysiloxane and the like. These above-mentioned oily components can be used alone or in combination of two or more.
In the present invention, among these, myristic acid glyceride, 2-ethylhexanoic acid triglyceride, lanolin, liquid paraffin, vaseline, paraffin microcrystalin wax, squalane, lauric acid, myristic acid, palmitic acid, linoleic acid, linolenic acid, isostearic acid. , Cetyl alcohol, stearyl alcohol, oleyl alcohol, cholesterol, cetyl octanoate, triglyceride octanate, isopropyl myristyrene, octyldodecyl myristate, cholesterol isostearate, POE sorbit fatty acid ester, peppermint oil, peppermint oil, coconut oil, rose It is preferable to use oil, menthol, cineol, eugenol, citral, citroneral, geraniol, pinen, limonene and dimethylpolysiloxane.

The following components can be further added to the formulation of the present invention, but the components are not limited thereto.
Pigments: Tar dyes specified in the Ordinance of the Ministry of Health and Welfare such as Yellow No. 4, Blue No. 1, Yellow No. 202, etc. Approved as food additives such as pigments in Appendix I and II, chlorophyll, riboflavin, crocin, safflower, anthraquinone, etc. Natural pigments, etc.
Vitamins; vitamin A, vitamin C, vitamin D, vitamin E, etc.
Others; bactericides, preservatives, other ingredients necessary for formulation, etc.

The pharmaceutical product of the present invention can be produced according to a conventional method by adding the optional ingredient to the essential ingredient as necessary, and can be in the form of cream, milky lotion, lotion or the like.

Next, the present invention will be described in detail with reference to examples.

Example 1
Add 2 liters of 50% (V / V) ethanol aqueous solution to 50 g of sagarame (dried product, shredded product), extract for 24 hours with occasional stirring, filter (No5C), evaporate, and then freeze-dry. bottom.

Example 2
Add 2 liters of 50% (V / V) ethanol aqueous solution to 50 g of Hibamata (Fucus vesiculosus) (dried product, shredded product), extract for 24 hours with occasional stirring, filter (No. 5C), and evaporate. This was lyophilized.

Example 3
Add 2 liters of 30% (V / V) ethanol aqueous solution to 30 g of black tea (dried product, shredded product), extract for 24 hours with occasional stirring, filter (No5C), evaporate, and then freeze-dry. bottom.

Example 4
Add 2 liters of 50% (V / V) ethanol aqueous solution to 30 g of Enmeisou (whole plant, dried product, shredded product), extract for 24 hours with occasional stirring, filter (No. 5C), evaporate, and then this. Was lyophilized.

Confirmation test After culturing the second-generation human foreskin-derived epidermal cells (Kurabou) in HuMedia-KG2 medium (without phenol red) so as to be 50-70% confluent, the calcium concentration was changed to 1.8 mM the day before. Examples were added to KG2 medium and cultured at 37 ° C. in a 5% CO2 incubator for 2 days.

<RNA extraction>
Extraction of Total RNA from cells was prepared using trypsin / EDTA and then using the illustra RNA Mini RNA Isolation Kit (GE Healthcare) according to the attached manual of GE Healthcare. The RNA concentration was calculated using NanoDrop1000 (Thermo SCIENTIFIC).

<RT reaction and real-time PCR>
An RT reaction was performed using 2.5 μg of Total RNA and MMLV Reverse Transcriptase RNase H- (Toyobo) according to the Toyobo recommended protocol (TOYOBO BIOCHEMICALS FOR LIFE SCIEENCE 2008/2009, page 1-42).
Real-time PCR was performed using Applied Biosystems 7500 real-time PCR System as follows. Gene expression by Comparative CT (△△ CT) method (n = 3) using the SYBR Green method (THUNDERBIRD SYBR qPCR Mix, Toyobo Co., Ltd.) and using the operation manual (Applied Biosystems) of the 7500 real-time PCR system. GAPDH was used as the internal standard. The target genes are filaggrin, loricrin, involucrin, and corneodesmosin (CDSN).

The result of the confirmation test is shown in FIG.

In addition, as a result of preparing an external preparation containing the examples and actually using it, improvements were observed in suppressing wrinkle formation, preventing rough skin, normalizing the texture, and promoting skin keratinization.

Examples 1 and 2 were allowed to act at an action concentration of 0.1%, Example 3 was allowed to act at an action concentration of 0.01%, and Example 4 was allowed to act at an action concentration of 0.005%. The gene expression level of desmosin (CDSN) was shown.

Claims (1)

A filaggrin production promoter containing at least one of the extract of Enmeisou and the extract of Hibamata as an active ingredient.

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