JP2018035145A - Filaggrin production promoter, involucrin production promoter, loricrin production promoter, histidase production promoter, and sprr1b production promoter - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a skin external preparation that promotes the production of filaggrin, involucrin, loricrin, histidase, and SPRR1B, normalizes a skin function, inhibits wrinkle formation, prevents a rough skin, normalizes a skin texture, and is effective against atopic dermatitis and the like.SOLUTION: The problem is solved by extraction of a Citrus junos extract, preferably a Citrus junos fruit with an alcohol aqueous solution and lyophilization of it.EFFECT: The external preparation has the effects of inhibiting wrinkle formation, preventing a rough skin, normalizing a skin texture, and promoting skin cornification.SELECTED DRAWING: Figure 1

Description

  The present invention promotes production of filaggrin, involucrin, loricrin, histidase, SPRR1B, promotes skin keratinization, maintains skin moisture, suppresses wrinkle formation, prevents rough skin, normalizes skin texture, atopic dermatitis, etc. The present invention relates to a composition for improving skin troubles.

The skin exists on the outermost side of the body, and has a role as a barrier against external stimuli such as bacteria. In the skin, epidermal cells keratinize from basal cells to spinous cells, granule cells, and even horny cells over about 4 weeks, in which intercellular lipids, NMF (natural moisturizing factor), The barrier function is achieved by forming a cornified envelope.
Although the formation of wrinkles, abnormal skin texture, rough skin, etc. occur due to the incomplete treatment of the stratum corneum with the aging of the skin and changes in the external environment, the reduction of the stratum corneum moisture content is an important factor.
Moisture retention in the keratin is related to natural moisturizing factors, but many parts of these natural moisturizing factors are related to amino acids generated by protein degradation.
The amino acid which is the main component of this natural moisturizing component is produced by the degradation of filaggrin derived from keratohyalin granules in the stratum corneum. This filaggrin is expressed as profilagrin in epidermal keratinocytes, immediately phosphorylated and accumulates in keratohyalin granules. It is then decomposed into filaggrin through dephosphorylation and hydrolysis, and then transferred to the stratum corneum, increasing the efficiency of keratin filament aggregation, 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 moisture retention of the skin, and that the synthetic power of filaggrin is reduced by conditions such as drying, and the amount of amino acids in the stratum corneum is reduced.

During the keratinization process of epidermal cells, when keratohyalin granules in the keratohyalin granules are keratinized, they are decomposed by the action of dephosphorylation and protease to release filaggrin protein, and the arginine residue of filaggrin Is modified to become a citrulline residue by an enzyme called peptidylarginine deiminase, and filaggrin is gradually detached from between keratin fibers and decomposed to produce an NMF component.
Moreover, trans-urocanic acid is produced by the action of histidase from histidine produced by the degradation of filaggrin, and protects the skin from ultraviolet rays.
Thus, the promotion of synthesis of filaggrin plays an important role in the retention of keratin water and the function of protecting against ultraviolet rays.

The epidermis has a protective function of protecting the skin from various stimuli from the outside by constantly creating new keratinocytes through the division and subsequent differentiation of keratinocytes. In particular, in the differentiation process of keratinocytes, proteins such as involucrin are expressed from the spinous layer to the granule layer, forming a confined envelope (CE), which is an insoluble cell membrane-like structure that wraps around keratinocytes. Contributes to the stability of the cytoskeleton and structure.
However, when the amount of involucrin produced in the epidermis is reduced due to various factors, the formation of a confined envelope (CE) becomes incomplete and keratinization is not performed normally. As a result, it is considered that the keratin barrier function and the skin moisturizing function are lowered, and skin symptoms such as rough skin and dry skin are exhibited.
For this reason, by increasing the production of involucrin in the epidermis of keratinocytes and promoting the formation of a cornified envelope to normalize keratinization, the skin barrier function associated with external stimuli such as drying and ultraviolet rays It is considered that various skin symptoms such as dry skin and rough skin can be prevented and improved.

Loricrin is a major component of the cornified cell envelope, and its expression begins with keratohyaline granules and is gradually incorporated into the peripheral zone. Involves in various stratum corneum functions, including wrinkles, abnormal texture, and rough skin.
Of course, it is also effective for atopic dermatitis, and can prevent and / or treat a skin disease that develops due to a decrease in the amount of free amino acids in the stratum corneum.

SPRR1B (small proline rich protein) is also a major component of the cornified cell envelope and is involved in normal keratinization and needs to be fully functioning.
In this way, by promoting the production of filaggrin, involucrin, loricrin, histidase, SPRR1B, the skin function is normalized, and an effective skin external preparation is obtained for suppressing wrinkle formation, preventing rough skin, normalizing skin texture, atopic dermatitis, etc. It is done.
Several substances that promote these productions are known, but no satisfactory one has been obtained.

Yuzu (Reiko, scientific name: Citrus junos) is an evergreen small Takagi tree of the genus Citrus. It is used as a spice and condiment in one of the citrus fruits. It is also used when taking a bath as Zushi-yu. Furthermore, it is also used for the purpose of antiseptic effect, bear improving agent, hyaluronic acid production accelerator and the like. (Patent Documents 1 to 3)

Japanese Patent Laid-Open No. 08-295621 JP 2001-072564 A JP 2001-158728 A

  The object of the present invention is to promote the production of filaggrin, involucrin, loricrin, histidase, SPRR1B, thereby normalizing skin function, and preventing skin wrinkle formation, preventing rough skin, normalizing skin condition, atopic dermatitis and the like. Is to get.

As a result of intensive studies by the present inventors, it has been found that a Yuzu extract achieves the above-mentioned purpose.
Yuzu has also been found to be particularly effective with fruit extracts.
It is preferable to perform extraction after performing processing such as shredding, drying, and pulverization in consideration of extraction efficiency by performing drying if necessary.
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 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 these hydrophilic organic solvents and water can be used.
In addition, when using the mixed solvent of the said water and a hydrophilic organic solvent, in the case of a lower alcohol, it is 3-20 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. On the other hand, it is preferable to add 3 to 15 parts by mass. In the case of a 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. Furthermore, in order to raise extraction efficiency, you may stir or homogenize in an extraction solvent. The extraction temperature is suitably about 5 ° C. to 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 single 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 a plurality of times.
If necessary, purification treatment such as deodorization and decolorization may be further added within a range that does not affect the effect, and concentration can be performed by a vacuum concentrator such as an evaporator or solvent removal by heating.
Further, this extract can be further purified using a synthetic adsorbent (Diaion HP20, Sephabies SP825, Amberlite XAD4, MCIgelCHP20P, etc.), dextran resin (Sephadex LH-20, etc.), ultrafiltration, etc. is there.

  The preparation of the present invention exhibits drug efficacy for oral, injection, and external use, but is preferably used as a skin external preparation. Skin external preparations include skin cosmetics, external quasi-drugs, and medical skin external preparations.

  In addition to the above ingredients, the preparations of the present invention include surfactants, oily ingredients, moisturizers, polymer compounds, ultraviolet absorbers, anti-inflammatory agents, bactericides, Antioxidants, sequestering agents, preservatives, vitamins, pigments, fragrances, water and the like can be blended.

  As the surfactant, any of anionic, cationic, nonionic, natural, and synthetic surfactants can be used, but it is preferable to use a nonionic surfactant in consideration of irritation to the skin. 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 sorbite fatty acid ester, polyoxyethylene alkyl ether, polyoxyethylene polyoxypropylene glycol. , Polyoxyethylene polyoxypropylene alkyl ether, polyethylene glycol fatty acid ester, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, alkylglycoside and the like.

  Examples of the oil component include fats and oils, waxes, hydrocarbons, higher fatty acids, higher alcohols, esters, essential oils, silicone oils, and the like. Examples of the fats and oils include soybean oil, nutka oil, jojoba oil, avocado oil, almond oil, olive oil, cacao oil, sesame oil, persic oil, castor oil, coconut oil, mink oil, beef fat, pork fat and the like, and these Hardened oil obtained by hydrogenation of natural fats and oils and synthetic triglycerides such as myristic acid glyceride and 2-ethylhexanoic acid triglyceride; waxes include, for example, carnauba wax, whale wax, beeswax, lanolin and the like; hydrocarbons For example, liquid paraffin, petrolatum, paraffin microcrystalline wax, ceresin, squalane, bristan etc .; higher fatty acids include, for example, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, linoleic acid, Linolenic acid, lanolinic acid, isostearic acid, etc .; Examples of the class alcohols include lauryl alcohol, cetyl alcohol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, 2-hexyldecanol, etc .; examples of the esters include cetyl octanoate, triglyceride octanoate, myristyl lactate, cetyl lactate, Examples of essential oils include mint oil and jasmine. Oil, pepper brain oil, cypress oil, spruce oil, ryu oil, turpentine oil, cinnamon oil, bergamot oil, mandarin oil, ginger oil, pine , Lavender oil, bay oil, clove oil, hiba oil, rose oil, eucalyptus oil, lemon oil, thyme oil, peppermint oil, rose oil, sage oil, menthol, cineole, eugenol, citral, citronellal, borneol, linalool, geraniol, Camphor, thymol, spirantol, pinene, limonene, terpene compounds, etc .; examples of silicone oils include dimethylpolysiloxane. These oily components described above 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, petrolatum, paraffin microcrystalline wax, squalane, lauric acid, myristic acid, palmitic acid, linoleic acid, linolenic acid, isostearic acid , Cetyl alcohol, stearyl alcohol, oleyl alcohol, cholesterol, cetyl octanoate, triglyceride octanoate, isopropyl myristate, octyldodecyl myristate, cholesterol isostearate, POE sorbite fatty acid ester, peppermint oil, spruce oil, cinnamon oil, rose Oil, menthol, cineol, eugenol, citral, citronellal, geraniol, pinene, limonene, dimethylpolysiloxane It is preferable to use.

The following components can be further added to the preparation of the present invention, but the components are not limited thereto.
Colors: Yellow color No. 4, Blue No. 1 and Yellow No. 202 are recognized as food additives such as tar color dyes I and II, chlorophyll, riboflavin, crocin, safflower, anthraquinone, etc. Natural pigments.
Vitamins; vitamin A, vitamin C, vitamin D, vitamin E, etc.
Others: bactericides, preservatives, other ingredients necessary for formulation.

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

EXAMPLES Next, an Example is given and this invention is demonstrated in detail.

Example 1
After crushing 100 g of yuzu fruit (blue yuzu), add 1 liter of 50% (V / V) ethanol aqueous solution, extract with stirring for 24 hours, filter (No5C), evaporate and freeze this Dried.

Confirmation test HuMedia- cultivated human foreskin-derived epidermis cells (Kurabo) in the 2nd passage in HuMedia-KG2 medium (without phenol red) so as to be 50-70% confluent, and the calcium concentration was changed to 1.8 mM the day before. Examples were added to KG2 medium and cultured in a 37 ° C., 5% CO 2 incubator for 2 days.

<Extraction of RNA>
Total RNA was extracted from the cells after exfoliation with trypsin / EDTA and then using 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>
Using 2.5 μg of total RNA, RT reaction was performed according to Toyobo recommended protocol (TOYOBO BIOCHEMICALS FOR LIFE SCIENCE 2008/2009, page 1-42) using MMLV Reverse Transcriptase RNase H- (Toyobo).
Real-time PCR was performed as follows using Applied Biosystems 7500 Real-Time PCR System. Using the SYBR Green method (THUNDERBIRD SYBR qPCR Mix, Toyobo Co., Ltd.) and the 7500 Real-Time PCR System operation manual (Applied Biosystems), gene expression comparison was performed by the Comparative CT (ΔΔCT) method (n = 3). GAPDH was used as an internal standard. The target genes are filaggrin, loricrin, involucrin, histidase, and SPRR1B.

  The results of the confirmation test are shown in FIGS.

  Moreover, the external preparation which mix | blended the Example was created, and as a result of actually using it, improvement was seen in wrinkle formation suppression, skin roughening prevention, skin normalization, and skin keratinization promotion.

The amount of gene expression when Example 1 was allowed to act at a working concentration of 0.2% was shown as 1 for the control. From left, gene expression levels of loricrin, filaggrin and histidase.

The amount of gene expression when Example 1 was allowed to act at a working concentration of 0.2% was shown as 1 for the control. From left, gene expression levels of involucrin and SPRR1B.

Claims (6)

  A filaggrin production promoter comprising one or more extracts of yuzu as an active ingredient.   An involucrin production promoter comprising one or more extracts of yuzu as an active ingredient.   A loricrin production promoter comprising one or more extracts of yuzu as an active ingredient.   A histidase production promoter comprising one or more extracts of yuzu as an active ingredient.   An SPRR1B production promoter comprising one or more extracts of yuzu as an active ingredient.   The filaggrin production promoter, involucrin production promoter, loricrin production promoter, histidase production promoter, SPRR1B production promoter according to any one of claims 1 to 5, wherein the yuzu extract is a yuzu fruit extract.