KLK5 mRNA expression promoter, KLK7 mRNA expression promoter, and SPINK5 mRNA expression promoter

Abstract

To provide a high-safety KLK5 mRNA expression promotor, a KLK7 mRNA expression promotor, an SPINK5 mRNA expression promotor, and a horny layer detachment improving agent or a horny layer detachment normalizing agent, and a skin cosmetic which have any effect of excellent KLK5 mRNA expression promoting effect, KLK7 mRNA expression promoting effect, and SPINK5 mRNA expression promoting effect.SOLUTION: A KLK5 mRNA expression promotor, a KLK7 mRNA expression promotor, and an SPINK5 mRNA expression promotor which contain olive extract as an active ingredient; a horny layer detachment improving agent or a horny layer detachment normalizing agent which contain at least one of the KLK5 mRNA expression promotor and the KLK7 mRNA expression promotor, and the SPINK5 mRNA expression promotor; and a skin cosmetic containing the horny layer detachment improving agent or the horny layer detachment normalizing agent.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a KLK5 mRNA expression promoter, a KLK7 mRNA expression promoter, a SPINK5 mRNA expression promoter, an agent for improving or normalizing stratum corneum desquamation, and a skin cosmetic. [Background technology]

[0002] The stratum corneum, the outermost layer of the epidermis, has a barrier function that protects the inside of the skin from various external stimuli. The stratum corneum is formed by keratinocytes (also called corneocytes). In healthy skin, keratinocytes are continuously produced by differentiation of keratinocytes (epidermal keratinocytes), and at the same time, old and unnecessary keratinocytes become dirt and peel off, thereby maintaining the stratum corneum at a certain thickness, and skin turnover is repeated. However, it is known that the barrier function of the skin is reduced by external factors such as dryness, ultraviolet rays, and chemicals, and internal factors such as oxidative stress and aging, and that these factors can cause abnormalities in skin turnover.

[0003] The stratum corneum is formed by adhesion between keratinocytes via an adhesive structure called corneodesmosome. Corneodesmosome is composed of multiple proteins including desmoglein 1 (DSG1) and desmocollin 1 (DSC1). Corneodesmosome is known to be cleaved by proteases such as kallikrein 5 (KLK5), a trypsin-like serine protease, and kallikrein 7 (KLK7), a chymotrypsin-like serine protease (see Non-Patent Documents 1 and 2).

[0004] In healthy skin, normal stratum corneum peeling occurs when corneodesmosomes are degraded by the serine protease. In normal stratum corneum peeling, the peeling is rarely recognized by the naked eye. However, when the activity of the serine protease is reduced due to skin troubles such as rough skin or aging, the decomposition of corneodesmosomes is delayed, causing the accumulation of keratinocytes, preventing smooth peeling, and the keratinocytes peel off in clumps, which are recognized as scales (see Non-Patent Document 3). Many skin diseases accompanied by abnormalities in the stratum corneum peeling function have been reported, such as senile xerosis, dandruff, ichthyosis vulgaris, sex-linked ichthyosis, and psoriasis (see Non-Patent Documents 4 to 6).

[0005] The serine protease activity in the epidermis is tightly regulated by serine protease inhibitors such as lympho-epithelial Kazal-type inhibitor (LEKTI) encoded by the serine peptidase inhibitor Kazal type 5 (SPINK5) gene. In patients with severe congenital ichthyosis-like erythroderma known as Netherton syndrome, the SPINK5 gene has been identified as the defective gene, and the activity of serine proteases such as KLK5 and KLK7 is increased, resulting in enhanced protein degradation.

[0006] Thus, it is important to promote the production of KLK5 and KLK7 in order to promote skin turnover.However, excessive production of KLK5 and KLK7 may cause abnormal desquamation.Therefore, in normal stratum corneum delamination, serine proteases such as KLK5 and KLK7 act on corneodesmosomes to decompose them, thereby promoting stratum corneum delamination, and at the same time, serine protease inhibitors such as LEKTI regulate and balance the serine protease activity, thereby maintaining the homeostasis of protease activity in the skin.

[0007] Skin conditions accompanied by abnormalities in the stratum corneum desquamation function as described above have the problem that they cause abnormalities in the skin surface shape, which greatly affects the appearance and significantly reduces the emotional quality of life. Therefore, improving the skin surface shape is important not only from a cosmetic perspective, but also for maintaining a healthy life.

[0008] Moisturizers and keratin care agents are commonly used to treat the skin surface. However, moisturizers have the problem that they take a relatively long time to take effect, making it difficult to feel the effects, and keratin care agents have the problem of being irritating. In addition, chemical peeling is a method of chemically dissolving the stratum corneum, and requires a doctor's diagnosis and skilled techniques.

[0009] Therefore, there is currently a strong demand for providing a technology that maintains the homeostasis of protease activity in the skin by regulating the balance between the activity of serine proteases such as KLK5 and KLK7 in the skin and the activity of serine proteases such as LEKTI, and thereby providing a highly safe stratum corneum desquamation improving agent or stratum corneum desquamation normalizing agent and skin cosmetic that can simply prevent, improve, or treat various skin symptoms caused by the external factors or internal factors. [Prior art documents] [Non-patent literature]

[0010] [Non-Patent Document 1] GM Yousef et al., Gene, 2000, Aug 22, 254(1-2), p.119-28 [Non-Patent Document 2] Cecile Caubet et al., J Invest Dermatol, 2004, 122, p.1235-44 [Non-Patent Document 3] AV Rawlings et al., Drug Discovery Today, 2008, 5(2), e127-e136 [Non-Patent Document 4] B Singh et al., Br J Dermatol, 2014, Oct;171(4), p.760-70 [Non-Patent Document 5] Y Kitajima et al., Dermatologica Sinica. 2015, 33(2), p.58-63 [Non-Patent Document 6] M Simon et al., Br J Dermatol. 2008, 159(1), p.77-85 Summary of the Invention [Problem to be solved by the invention]

[0011] The present invention aims to solve the above-mentioned problems in the prior art and to achieve the following objects. That is, an object of the present invention is to provide a KLK5 mRNA expression promoter that has excellent KLK5 mRNA expression-promoting activity and is highly safe. Another object of the present invention is to provide a KLK7 mRNA expression promoter that has excellent KLK7 mRNA expression-promoting activity and is highly safe. Another object of the present invention is to provide a SPINK5 mRNA expression promoter that has an excellent SPINK5 mRNA expression-promoting effect and is highly safe. Another object of the present invention is to provide a stratum corneum delamination improving agent or stratum corneum delamination normalizing agent which has at least one of an excellent KLK5 mRNA expression promoting effect and an excellent SPINK5 mRNA expression promoting effect, and which is highly safe, as well as a skin cosmetic preparation containing the stratum corneum delamination improving agent or stratum corneum delamination normalizing agent. [Means for solving the problem]

[0012] The means for solving the above problems are as follows. <1> The present invention is a KLK5 mRNA expression promoter characterized by containing olive extract as an active ingredient. <2> It is a KLK7 mRNA expression promoter characterized by containing olive extract as an active ingredient. <3> It is a SPINK5 mRNA expression promoter characterized by containing olive extract as an active ingredient. <4> At least any one of the KLK5 mRNA expression promoter described in <1> and the KLK7 mRNA expression promoter described in <2>, and the SPINK5 mRNA expression promoter described in <3>, is a horny layer peeling improver or horny layer peeling normalizer characterized by containing them. <5> It is a skin cosmetic characterized by containing the horny layer peeling improver or horny layer peeling normalizer described in <4>.

Effects of the Invention

[0013] According to the present invention, the above-mentioned various problems in the prior art can be solved, the above-mentioned object can be achieved, a KLK5 mRNA expression promoter having an excellent KLK5 mRNA expression promoting effect and high safety can be provided. Further, the present invention can provide a KLK7 mRNA expression promoter having an excellent KLK7 mRNA expression promoting effect and high safety. Further, the present invention can provide a SPINK5 mRNA expression promoter having an excellent SPINK5 mRNA expression promoting effect and high safety. Further, the present invention has at least any one of an excellent KLK5 mRNA expression promoting effect and an excellent KLK7 mRNA expression promoting effect, and an excellent SPINK5 mRNA expression promoting effect, and can provide a highly safe horny layer peeling improver or horny layer peeling normalizer, and a skin cosmetic containing the horny layer peeling improver or horny layer peeling normalizer.

Modes for Carrying Out the Invention

[0014] (KLK5 mRNA Expression Promoter, KLK7 mRNA Expression Promoter, and SPINK5 mRNA Expression Promoter) <KLK5 mRNA Expression Promoter> The KLK5 mRNA expression promoter of the present invention contains olive extract as an active ingredient and, if necessary, further contains other components.

[0015] The KLK5 gene is a gene encoding kallikrein 5 (KLK5), a trypsin-like serine protease expressed in the skin. KLK5 is an enzyme involved in the adhesion between stratum corneum cells in the skin and promotes stratum corneum desquamation. Therefore, the KLK5 mRNA expression promoter is suitably used for promoting stratum corneum desquamation. Although the details of the substance in the olive extract that exhibits the KLK5 mRNA expression promoting effect are unknown, the fact that the olive extract has such an excellent effect and is useful as a KLK5 mRNA expression promoter has not been known at all in the past and is a new finding by the present inventor.

[0016] <KLK7 mRNA Expression Promoter> The KLK7 mRNA expression promoter of the present invention contains olive extract as an active ingredient and, if necessary, further contains other components.

[0017] The KLK7 gene is a gene encoding kallikrein 7 (KLK7), a chymotrypsin-like serine protease expressed in the skin. KLK7 is an enzyme involved in the adhesion between stratum corneum cells in the skin and promotes stratum corneum desquamation. Therefore, the KLK7 mRNA expression promoter is suitably used for promoting stratum corneum desquamation. Although the details of the substance in the olive extract that exhibits the KLK7 mRNA expression promoting effect are unknown, the fact that the olive extract has such an excellent effect and is useful as a KLK7 mRNA expression promoter has not been known at all in the past and is a new finding by the present inventor.

[0018] <SPINK5 mRNA Expression Promoter> The SPINK5 mRNA expression promoter of the present invention contains olive extract as an active ingredient and, if necessary, further contains other components.

[0019] The SPINK5 gene encodes a serine protease inhibitor (LEKTI) expressed in the skin, which regulates stratum corneum desquamation by suppressing the activity of KLK5 and KLK7. Although the details of the substance that the olive extract possesses in promoting SPINK5 mRNA expression are unknown, the fact that the olive extract has such an excellent effect and is useful as a SPINK5 mRNA expression promoter was not previously known, and is a new discovery by the present inventors.

[0020] <<Olive extract>> Olives (scientific name: Olea european ) is a member of the Oleaceae family ( Oleaceae )Olive genus( Olea Olives are an evergreen tree belonging to the olive family. The oil extracted from the fruit is used for food, medicine, and cosmetics. Olives are cultivated in the Mediterranean region and in Asia, including China, and are easily available in these areas.

[0021] The olive extract to be used may be one prepared from the part of the olive to be used as the extraction raw material, or a commercially available product may be used.

[0022] The part of the olive to be extracted is not particularly limited and can be appropriately selected according to the purpose, and examples thereof include above-ground parts such as leaves, branches, branches and leaves, bark, trunks, flowers, buds, fruits, pericarp, seeds, and seed coats; and underground parts such as roots and rhizomes. These may be used alone or in combination of two or more. Among these, the part of the olive to be extracted is preferably the leaves, since it is excellent in at least one of the actions of promoting KLK5 mRNA expression, promoting KLK7 mRNA expression, and promoting SPINK5 mRNA expression.

[0023] The size of the olive extract portion is not particularly limited and can be appropriately selected depending on the purpose. For example, the size may be the size as it is when harvested, the desired size after cutting, or the size after pulverization (powder).

[0024] The state of the olives to be extracted is not particularly limited as long as it is in a state that allows extraction, and can be appropriately selected depending on the purpose, and examples include the state as picked (raw), dried, crushed, juiced, etc. Among these, the state of the olives to be extracted is preferably the state as picked, crushed, or juiced, and more preferably the state as picked or crushed.

[0025] The method for drying the extracted olive parts is not particularly limited and can be appropriately selected depending on the purpose. Examples of the method include drying in the sun and drying using a commonly used dryer.

[0026] The method for pulverizing the extracted olive part is not particularly limited and can be appropriately selected depending on the purpose. For example, the method includes pulverizing the olive part using a mixer, a sugar mill, a power mill, a jet mill, an impact crusher, etc.

[0027] The method for converting the extracted part of the olive into the juice is not particularly limited and can be appropriately selected depending on the purpose. For example, squeezing or the like can be used.

[0028] The olive extract can be easily obtained by a method generally used for plant extraction. The form of the olive extract is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include a diluted product, a concentrated product, a dried product, a crude product, and a purified product of the olive extract.

[0029] The method for extracting the olives is not particularly limited and may be appropriately selected depending on the purpose. For example, an extract may be obtained by placing the part of the olive to be extracted, which is the raw material for extraction, in a treatment tank filled with an extraction solvent, stirring appropriately as necessary to elute the soluble components, and then filtering to remove the extraction residue. In addition, olives may be used as an extraction material after being pretreated, such as by degreasing, with a non-polar solvent such as hexane or benzene. By carrying out pretreatment such as degreasing, the olives can be efficiently extracted with a polar solvent.

[0030] The extraction conditions such as extraction time and extraction temperature, and the extraction solvent and the amount used thereof in the extraction of the olives are not particularly limited and can be appropriately selected depending on the purpose.

[0031] The extracting solvent is not particularly limited and can be appropriately selected according to the purpose, but a polar solvent is preferable. The components that exhibit the KLK5 mRNA expression promoting effect, the KLK7 mRNA expression promoting effect, and the SPINK5 mRNA expression promoting effect contained in the olive can be easily extracted by an extraction process using a polar solvent as the extracting solvent.

[0032] The polar solvent is not particularly limited and can be appropriately selected depending on the purpose. Examples of the polar solvent include water, a hydrophilic organic solvent, and a mixed solvent thereof.

[0033] The water is not particularly limited and can be appropriately selected depending on the purpose. Examples of the water include pure water, tap water, well water, mineral water, hot spring water, spring water, and fresh water. In addition, the water may be one that has been subjected to various treatments. Examples of the treatments that can be applied to the water include heating, sterilization, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, etc. Therefore, the water that can be used as the extraction solvent may be hot water, purified water, ion exchange water, physiological saline, phosphate buffer, phosphate buffered physiological saline, etc.

[0034] The hydrophilic solvent is not particularly limited and can be appropriately selected depending on the purpose. Examples of the hydrophilic 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; and polyhydric alcohols having 2 to 5 carbon atoms, such as 1,3-butylene glycol, propylene glycol, and glycerin.

[0035] The mixed solvent of water and a hydrophilic solvent is not particularly limited and can be appropriately selected depending on the purpose. When a lower alcohol is used, it is preferable to add 1 part by mass to 90 parts by mass per 10 parts by mass of water, when a lower aliphatic ketone is used, it is preferable to add 1 part by mass to 40 parts by mass per 10 parts by mass of water, and when a polyhydric alcohol is used, it is preferable to add 1 part by mass to 90 parts by mass per 10 parts by mass of water.

[0036] The extraction solvent is preferably used at a temperature between room temperature and reflux.

[0037] The olive extraction process is not particularly limited as long as it can dissolve the soluble components contained in the olives in the extraction solvent, and can be appropriately selected from among conventional methods. There is no need to adopt a special extraction method when extracting the olives, and any device can be used at room temperature or under reflux heating.

[0038] As a specific example of the olive extraction process, the olive extraction part as the extraction raw material is put into a processing tank filled with the extraction solvent, and the soluble components are dissolved while stirring occasionally. In this case, the extraction conditions can be appropriately adjusted according to the state of the olive extraction part, the extraction solvent, etc. For example, when water is used as the extraction solvent, the amount of the extraction solvent is usually about 1 to 1,000 times the amount of the extraction raw material (mass ratio), the extraction time is usually about 30 minutes to 2 hours, and the extraction temperature is usually about 40 ° C to 90 ° C. Also, for example, when a mixed solvent of water and ethanol is used as the extraction solvent, the amount of the extraction solvent is usually about 1 to 100 times the amount of the extraction raw material (mass ratio), the extraction time is usually about 30 minutes to 2 hours, and the extraction temperature is usually about 40 ° C to 90 ° C.

[0039] After soluble components are eluted from the olives by the extraction treatment, the olive extract can be obtained by filtering to remove the extraction residue.

[0040] The obtained olive extract may be subjected to treatments such as dilution, concentration, drying, purification, etc. according to conventional methods to obtain a diluted product, concentrate, dry product, crude product, purified product, etc. of the olive extract.

[0041] The olive extract may be purified for the purposes of decolorization, deodorization, activity improvement, etc., as long as the effects of the present invention can be obtained, but it may also be used in an unpurified form.

[0042] When purifying the olive extract for the purpose of decolorization, deodorization, improving physiological activity, etc., the purification method is not particularly limited and can be appropriately selected depending on the purpose. For example, activated carbon treatment, resin adsorption treatment, treatment using ion exchange resin, liquid-liquid countercurrent distribution, etc. can be mentioned.

[0043] The olive extract exhibits at least any one of an excellent KLK5 mRNA expression promoting effect, an excellent KLK7 mRNA expression promoting effect, and an excellent SPINK5 mRNA expression promoting effect, and can therefore be suitably used as the KLK5 mRNA expression promoter, the KLK7 mRNA expression promoter, or an active ingredient of the SPINK5 mRNA expression promoter, or as a stratum corneum desquamation improving agent, stratum corneum desquamation normalizing agent, or a compounding ingredient of a skin cosmetic preparation, as described below.

[0044] The olive extract can be used as it is as the active ingredient of the KLK5 mRNA expression promoter, the KLK7 mRNA expression promoter, or the SPINK5 mRNA expression promoter, or as a compounding ingredient of the stratum corneum peeling improving agent, the stratum corneum peeling normalizing agent, or the skin cosmetic described below, but can also be used after being formulated according to a conventional method.When formulating, dextrin, cyclodextrin, or other pharmacopoeia-acceptable carriers or other optional auxiliary agents may be added to facilitate storage and handling.By formulating, it can be made into any dosage form such as powder, granules, or tablets.

[0045] The content of the olive extract in the KLK5 mRNA expression promoter, the KLK7 mRNA expression promoter, or the SPINK5 mRNA expression promoter is not particularly limited and can be appropriately adjusted depending on the physiological activity of the olive extract, etc. The KLK5 mRNA expression promoter, the KLK7 mRNA expression promoter, or the SPINK5 mRNA expression promoter may consist of the olive extract alone.

[0046] <<Other ingredients>> The other components in the KLK5 mRNA expression promoter, the KLK7 mRNA expression promoter, or the SPINK5 mRNA expression promoter are not particularly limited and can be appropriately selected according to the form of use of the KLK5 mRNA expression promoter, the KLK7 mRNA expression promoter, or the SPINK5 mRNA expression promoter. For example, excipients, moisture-proofing agents, preservatives, strengthening agents, thickeners, emulsifiers, antioxidants, sweeteners, acidulants, seasonings, colorants, fragrances, whitening agents, moisturizers, oily components, UV absorbers, surfactants, thickeners, alcohols, powder components, colorants, aqueous components, water, skin nutrients, etc. may be mentioned. These may be used alone or in combination of two or more. In addition, if necessary, a substance other than the olive extract having at least one of the actions of KLK5 mRNA expression promotion, KLK7 mRNA expression promotion, and SPINK5 mRNA expression promotion may be added. The content of the other components is not particularly limited as long as it does not impair the effects of the present invention, and can be appropriately selected depending on the purpose.

[0047] -Applications- The KLK5 mRNA expression promoter of the present invention has an excellent KLK5 mRNA expression promoting effect, the KLK7 mRNA expression promoter of the present invention has an excellent KLK7 mRNA expression promoting effect, and the SPINK5 mRNA expression promoter of the present invention has an excellent SPINK5 mRNA expression promoting effect, and therefore can be suitably used as a medicine, quasi-drug, cosmetic, etc. used in the prevention, amelioration, or treatment of skin symptoms accompanied by abnormalities in stratum corneum desquamation function due to external factors such as dryness, ultraviolet rays, and chemicals, and internal factors such as oxidative stress and aging (e.g., senile xerosis, dandruff, ichthyosis vulgaris, sex-linked ichthyosis, psoriasis, etc.), and the amount of incorporation, method of use, and dosage form can be appropriately selected depending on the purpose of use.

[0048] The method of use is not particularly limited and can be appropriately selected depending on the purpose, but external application is preferred because it acts easily on the stratum corneum of the skin.

[0049] The dosage form is not particularly limited and can be appropriately selected depending on the purpose. Examples of the dosage form include lotion, milky lotion, cream, ointment, beauty essence, lotion, pack, jelly, lip balm, lipstick, foundation, bath additives, soap, body shampoo, astringent, hair tonic, hair cream, hair liquid, pomade, shampoo, and rinse.

[0050] In addition, the KLK5 mRNA expression promoter, the KLK7 mRNA expression promoter, or the SPINK5 mRNA expression promoter can also be used as a reagent for research into the mechanism of action of the KLK5 mRNA expression promoting effect, the KLK7 mRNA expression promoting effect, or the SPINK5 mRNA expression promoting effect.

[0051] The KLK5 mRNA expression promoter of the present invention, the KLK7 mRNA expression promoter of the present invention, and the SPINK5 mRNA expression promoter of the present invention are preferably applied to humans, but can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as their respective functional effects are achieved.

[0052] In addition, the KLK5 mRNA expression promoter, the KLK7 mRNA expression promoter, and the SPINK5 mRNA expression promoter can improve skin turnover and thereby improve skin barrier function based on the activity of KLK5, KLK7, and LEKTI, the production of which is promoted by the KLK5 mRNA expression promoting effect, the KLK7 mRNA expression promoting effect, or the SPINK5 mRNA expression promoting effect, and can therefore be suitably used as epidermal turnover improvers and epidermal barrier function improvers, etc. Furthermore, the KLK5 mRNA expression promoter and the KLK7 mRNA expression promoter can degrade corneodesmosomes by KLK5 and KLK7, the production of which is promoted by the KLK5 mRNA expression promoting effect and the KLK7 mRNA expression promoting effect, and therefore can be suitably used as a corneodesmosome degradation normalizer, and the SPINK5 mRNA expression promoter can promote the production of LEKTI by its SPINK5 mRNA expression promoting effect, and therefore can be suitably used as a LEKTI production promoter.

[0053] (Stratum corneum peeling improvement agent or stratum corneum peeling normalization agent) The stratum corneum desquamation improving agent or stratum corneum desquamation normalizing agent of the present invention contains at least one of the KLK5 mRNA expression promoting agent of the present invention and the KLK7 mRNA expression promoting agent of the present invention, and the SPINK5 mRNA expression promoting agent of the present invention, and if necessary, further contains other ingredients.

[0054] As used herein, "stratum corneum desquamation" refers to a state in which adhesion in the adhesive structures (corneodesmosomes) between stratum corneum cells weakens, causing old stratum corneum cells to peel off from the skin surface.

[0055] The olive extract, which is an active ingredient of the KLK5 mRNA expression promoter, the KLK7 mRNA expression promoter, and the SPINK5 mRNA expression promoter, has excellent KLK5 mRNA expression promoting effect, excellent KLK7 mRNA expression promoting effect, and excellent SPINK5 mRNA expression promoting effect.

[0056] Therefore, in this specification, "improving stratum corneum desquamation" means promoting stratum corneum desquamation based on at least one of the effects of promoting KLK5 mRNA expression and KLK7 mRNA expression, while at the same time achieving a balance so that stratum corneum desquamation alone is not activated based on the effect of promoting SPINK5 mRNA expression, thereby bringing stratum corneum desquamation closer to the state of stratum corneum desquamation in healthy skin.

[0057] In addition, in this specification, "normalization of stratum corneum desquamation" refers to promoting stratum corneum desquamation based on at least one of the effects of promoting KLK5 mRNA expression and KLK7 mRNA expression, while at the same time achieving a balance so that only stratum corneum desquamation is not activated based on the effect of promoting SPINK5 mRNA expression, thereby restoring stratum corneum desquamation to the state it is in healthy skin.

[0058] Therefore, the stratum corneum delamination improving agent or stratum corneum delamination normalizing agent of the present invention is useful for preventing, improving, or treating abnormalities in the skin surface shape (e.g., dryness, flaking, peeling, bumps, rough texture, etc.) resulting from the accumulation of stratum corneum cells observed in skin conditions accompanied by abnormalities in the stratum corneum delamination function due to external factors such as dryness, ultraviolet rays, and chemicals, and internal factors such as oxidative stress and aging (e.g., senile xerosis, dandruff, ichthyosis vulgaris, X-linked ichthyosis, psoriasis, etc.).

[0059] Although the details of the substances that exhibit the KLK5 mRNA expression promoting effect, KLK7 mRNA expression promoting effect, and SPINK5 mRNA expression promoting effect of the olive extract are unknown, the fact that the olive extract has such excellent effects and is useful as a stratum corneum desquamation improving agent or a stratum corneum desquamation normalizing agent has never been known conventionally and is a new finding by the present inventor.

[0060] <KLK5 mRNA Expression Promoter, KLK7 mRNA Expression Promoter, and SPINK5 mRNA Expression Promoter> The KLK5 mRNA expression promoter, the KLK7 mRNA expression promoter, and the SPINK5 mRNA expression promoter in the stratum corneum desquamation improving agent or the stratum corneum desquamation normalizing agent are the KLK5 mRNA expression promoter of the present invention, the KLK7 mRNA expression promoter of the present invention, and the SPINK5 mRNA expression promoter of the present invention as described above, respectively.

[0061] The content or total content of the KLK5 mRNA expression promoter, the KLK7 mRNA expression promoter, and the SPINK5 mRNA expression promoter in the stratum corneum desquamation improving agent or the stratum corneum desquamation normalizing agent is not particularly limited and can be appropriately adjusted according to the physiological activity of the olive extract as the active ingredient. Further, the stratum corneum desquamation improving agent or the stratum corneum desquamation normalizing agent may consist only of the olive extract.

[0062] <Other Components> The other components are not particularly limited, and can be appropriately selected according to the form of use of the stratum corneum peeling improving agent or the stratum corneum peeling normalizing agent, and can include, for example, excipients, moisture-proofing agents, preservatives, strengthening agents, thickeners, emulsifiers, antioxidants, sweeteners, acidulants, seasonings, coloring agents, fragrances, whitening agents, moisturizing agents, oily components, UV absorbers, surfactants, thickeners, alcohols, powder components, coloring agents, aqueous components, water, skin nutrients, etc. These can be used alone or in combination of two or more. In addition, if necessary, a substance other than the olive extract that has at least one of the following actions of promoting KLK5 mRNA expression, promoting KLK7 mRNA expression, and promoting SPINK5 mRNA expression can be added. The content of the other components is not particularly limited as long as it does not impair the effects of the present invention, and can be appropriately selected depending on the purpose.

[0063] -Applications- The stratum corneum peeling improving agent or stratum corneum peeling normalizing agent of the present invention has an excellent stratum corneum peeling improving action or stratum corneum peeling normalizing action based on at least one of the excellent KLK5 mRNA expression promoting action and the excellent KLK7 mRNA expression promoting action of the olive extract, which is the active ingredient of the KLK5 mRNA expression promoting agent, the KLK7 mRNA expression promoting agent, and the SPINK5 mRNA expression promoting agent, and the excellent SPINK5 mRNA expression promoting action, and is highly safe, so that it can be suitably used as a medicine, quasi-drug, cosmetic, etc. used for preventing, treating, or improving skin symptoms (e.g., senile xerosis, dandruff, ichthyosis vulgaris, sex-linked hereditary ichthyosis, psoriasis, etc.) accompanied by abnormality of stratum corneum peeling function due to external factors such as dryness, ultraviolet rays, chemicals, etc., and internal factors such as oxidative stress and aging, and can be suitably used as the skin cosmetic of the present invention described later. In addition, its blending amount, usage, and dosage form can be appropriately selected according to its purpose of use.

[0064] The method of use is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include external use. Among these, external use is preferred.

[0065] The dosage form is not particularly limited and can be appropriately selected depending on the purpose. Examples of the dosage form include lotion, milky lotion, cream, ointment, beauty essence, lotion, pack, jelly, lip balm, lipstick, foundation, bath additives, soap, body shampoo, astringent, hair tonic, hair cream, hair liquid, pomade, shampoo, and rinse.

[0066] The stratum corneum delamination improving agent or the stratum corneum delamination normalizing agent can also be used as a reagent for research into the mechanism of action of stratum corneum delamination.

[0067] The stratum corneum delamination improving agent or stratum corneum delamination normalizing agent of the present invention is preferably applied to humans, but can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) so long as the respective action effects are exerted.

[0068] (Skin cosmetics) The skin cosmetic of the present invention contains the stratum corneum desquamation improving agent or stratum corneum desquamation normalizing agent of the present invention, and further contains other ingredients as necessary.

[0069] <Stratum corneum peeling improvement agent or stratum corneum peeling normalization agent> The stratum corneum delamination improving agent or the stratum corneum delamination normalizing agent in the skin cosmetic preparation is the stratum corneum delamination improving agent or stratum corneum delamination normalizing agent of the present invention described above.

[0070] The content of the stratum corneum peeling improving agent or the stratum corneum peeling normalizing agent in the skin cosmetic is not particularly limited and can be appropriately selected according to purpose, but is preferably 0.1 volume % or more, more preferably 0.25 volume % or more, based on the total amount of the skin cosmetic.The content of the stratum corneum peeling improving agent or the stratum corneum peeling normalizing agent is preferably as high as possible, and its upper limit is not particularly limited and can be appropriately selected according to purpose.The skin cosmetic may be the stratum corneum peeling improving agent or the stratum corneum peeling normalizing agent itself.

[0071] <Other ingredients> The other components in the skin cosmetic preparation are not particularly limited and can be appropriately selected according to the purpose from among main ingredients, auxiliary ingredients, or other ingredients used in the production of ordinary skin cosmetic preparations, and examples thereof include astringents, bactericides, antibacterial agents, whitening agents, ultraviolet absorbers, moisturizers, cell activators, anti-inflammatory agents, antiallergic agents, antioxidants, active oxygen removers, oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, fragrances, etc. These may be used alone or in combination of two or more. The content of the other ingredients in the skin cosmetic is not particularly limited as long as it does not impair the effects of the present invention, and can be appropriately selected depending on the purpose.

[0072] The skin cosmetic of the present invention can be used on a daily basis, and due to the action of the active ingredient, the olive extract, it can extremely effectively exert various physiologically active actions such as promoting KLK5 mRNA expression, promoting KLK7 mRNA expression, and promoting SPINK5 mRNA expression, thereby improving or normalizing stratum corneum peeling and making it suitable for use in preventing, improving, or treating various skin conditions.

[0073] The type of the skin cosmetic is not particularly limited and can be appropriately selected depending on the purpose. Examples of the skin cosmetic include lotion, milky lotion, cream, ointment, beauty essence, lotion, pack, jelly, lip balm, lipstick, foundation, bath additive, soap, body shampoo, astringent, hair tonic, hair cream, hair liquid, pomade, shampoo, and rinse.

[0074] The skin cosmetic preparations are suitable for use on humans, but can also be used on animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as the respective functional effects are achieved. EXAMPLES

[0075] The present invention will be specifically described below with reference to Production Examples and Test Examples, but the present invention is not limited to these Production Examples and Test Examples.

[0076] (Production Example 1) 1,500 mL of 50% by volume ethanol aqueous solution was added to 100 g of dried olive leaves, and extraction was carried out at about 85°C for 2 hours using a reflux extractor, followed by hot filtration to remove the extraction residue, obtaining an extract. The obtained extract was dried to obtain about 20 g of olive extract.

[0077] (Test Example 1) The olive extract obtained in Production Example 1 was added to a 50 volume% aqueous 1,3-butylene glycol solution so that the solids concentration of the olive extract was 1.5 mass%. This was used as a test sample, and the KLK5 mRNA expression promoting effect, KLK7 mRNA expression promoting effect, and SPINK5 mRNA expression promoting effect were tested using the test methods described below.

[0078] Normal human epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) at 37°C under 5% CO2, and then the cells were harvested by trypsinization. The number of cells harvested using KGM was adjusted to 30 × 10 per 35 mm dish. 4 The cells were seeded at 2 mL per well and cultured overnight at 37°C under 5% CO2. After the culture, the culture medium was replaced with a medium (KBM) containing no growth factors and cultured at 37°C under 5% CO2 for 24 hours. After the culture, the culture medium was removed, and 2 mL of the test sample dissolved in KBM at the required concentration or KBM without the test sample was added to each petri dish and cultured at 37°C under 5% CO2 for 24 hours. After the culture, the culture medium was removed, and total RNA was extracted using ISOGEN II (manufactured by Nippon Gene Co., Ltd.), and the amount of RNA was calculated from the absorbance at a wavelength of 260 nm, and the total RNA was adjusted to 200 ng / μL.

[0079] Using this total RNA as a template, the expression levels of each of KLK5, KLK7, SPINK5, and GAPDH (an internal standard) mRNA were measured. Detection was performed using a real-time PCR device (Thermal Cycler Dice® Real Time System III (manufactured by Takara Bio Inc.)) with PrimeScript TM A two-step real-time RT-PCR reaction was performed using RT Master Mix (Perfect Real Time) (Takara Bio Inc.) and TB Green (registered trademark) Fast qPCR Mix (Takara Bio Inc.). The expression levels of KLK5, KLK7, and SPINK5 mRNA when the test sample was added or when the test sample was not added were corrected with the expression level of GAPDH mRNA, and the corrected values ​​were calculated. From the corrected values ​​obtained, the KLK5 mRNA expression promotion rate (%) was calculated using the following formula 1, the KLK7 mRNA expression promotion rate (%) was calculated using the following formula 2, and the SPINK5 mRNA expression promotion rate (%) was calculated using the following formula 3. The results are shown in Tables 1 to 3. <Expression 1> KLK5 mRNA expression promotion rate (%) = A / B × 100 A: Corrected value of KLK5 mRNA expression level when test sample was added B: Corrected value of KLK5 mRNA expression level when no test sample was added <Expression 2> KLK7 mRNA expression promotion rate (%) = C / D × 100 C: Corrected value of KLK7 mRNA expression level when test sample was added D: Corrected value of KLK7 mRNA expression level when no test sample was added <Expression 3> SPINK5 mRNA expression promotion rate (%) = E / F × 100 E: Corrected value of SPINK5 mRNA expression level when test sample was added F: Corrected value of SPINK5 mRNA expression level when no test sample was added

[0080] [Table 1]

[0081] [Table 2]

[0082] [Table 3]

[0083] The results in Tables 1 to 3 confirmed that the olive extract had excellent KLK5 mRNA expression promoting effect, KLK7 mRNA expression promoting effect, and SPINK5 mRNA expression promoting effect. [Industrial Applicability]

[0084] The KLK5 mRNA expression promoter of the present invention has an excellent KLK5 mRNA expression promoting effect, the KLK7 mRNA expression promoter of the present invention has an excellent KLK7 mRNA expression promoting effect, and the SPINK5 mRNA expression promoter of the present invention has an excellent SPINK5 mRNA expression promoting effect, and therefore can be suitably used as a pharmaceutical, quasi-drug, cosmetic, etc. The stratum corneum delamination improving agent or stratum corneum delamination normalizing agent of the present invention has at least one of an excellent KLK5 mRNA expression promoting effect and an excellent KLK7 mRNA expression promoting effect, as well as an excellent SPINK5 mRNA expression promoting effect, and therefore can be suitably used as a pharmaceutical, quasi-drug, cosmetic, etc., and can be particularly suitably used as a skin cosmetic. Since the skin cosmetic preparation of the present invention contains the stratum corneum delamination improving agent or stratum corneum delamination normalizing agent of the present invention, it is capable of improving or normalizing stratum corneum delamination and can be suitably used for preventing, improving, or treating various skin conditions.

Claims

1. A KLK5 mRNA expression promoter comprising an olive extract as an active ingredient.

2. A KLK7 mRNA expression promoter comprising an olive extract as an active ingredient.

3. A SPINK5 mRNA expression promoter comprising an olive extract as an active ingredient.

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