阅读说明：本技术 评价或选择感觉刺激降低剂的方法 (Methods of evaluating or selecting sensory stimulation reducing agents ) 是由 二之宫理恵 西岛贵史 佐藤纪子 于 2018-11-29 设计创作，主要内容包括：本发明提供一种对降低因对羟基苯甲酸酯类所致的感觉刺激的感觉刺激降低剂进行评价或选择的方法、及降低因对羟基苯甲酸酯类所致的感觉刺激的感觉刺激降低剂。本发明的对降低因对羟基苯甲酸酯类所致的感觉刺激的感觉刺激降低剂的评价或选择方法包括以下的(1)～(3)的工序：(1)使受试物质与可表达CES1的细胞接触的工序；(2)测定该细胞中的CES1的表达的工序；及(3)基于(2)中所测定的结果,将促进CES1的表达的受试物质评价为降低因对羟基苯甲酸酯类所致的感觉刺激的感觉刺激降低剂的工序。(The present invention provides a method for evaluating or selecting a sensory stimulation reducing agent for reducing sensory stimulation due to parabens, and a sensory stimulation reducing agent for reducing sensory stimulation due to parabens. The method for evaluating or selecting a sensory stimulation reducing agent for reducing sensory stimulation due to parabens of the present invention comprises the following steps (1) to (3): (1) a step of contacting a test substance with a cell capable of expressing CES 1; (2) measuring the expression of CES1 in the cell; and (3) evaluating the test substance that promotes the expression of CES1 as a sensory stimulation reducing agent that reduces sensory stimulation by parabens based on the results of the measurement in (2).)

1. A method for evaluating or selecting a sensory stimulation reducing agent for reducing sensory stimulation due to parabens, wherein,

the method comprises the following steps (1) to (3):

(1) a step of contacting a test substance with a cell capable of expressing CES 1;

(2) measuring the expression of CES1 in the cell; and

(3) and (3) evaluating a test substance that promotes the expression of CES1 as a sensory stimulation reducing agent that reduces sensory stimulation by parabens based on the result of the measurement in (2).

2. The method of claim 1, wherein,

the parabens are selected from one or more of methyl paraben, sodium methyl paraben, ethyl paraben, butyl paraben, propyl paraben, isobutyl paraben, isopropyl paraben and benzyl paraben.

3. The method of claim 1 or 2,

cells that express CES1 are cells derived from human epidermal tissue.

4. A sensory stimulation reducing agent, wherein,

sulforaphane, or a plant or an extract thereof containing sulforaphane or its glycoside as an active ingredient.

5. The sensory stimulation reducing agent according to claim 4, wherein,

the sensory stimulation is a stimulation sensation caused by parabens.

6. An agent for promoting CES1 expression, wherein,

sulforaphane, or a plant or an extract thereof containing sulforaphane or its glycoside as an active ingredient.

7. The agent according to any one of claims 4 to 6,

the plant containing sulforaphane or its glycoside is germinated broccoli.

8. Use of sulforaphane, or a plant or an extract thereof containing sulforaphane or a glycoside thereof for the manufacture of a sensory stimulation reducing agent.

9. Use of sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof for the manufacture of a CES1 expression promoter.

10. Sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof, for reducing sensory stimulation.

11. Sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof, for promoting expression of CES 1.

12. Non-therapeutic use of sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof, for reducing sensory stimulation.

13. Non-therapeutic use of sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof, for promoting expression of CES 1.

14. A method of reducing sensory stimulation, wherein,

sulforaphane, or a plant or extract thereof containing sulforaphane or its glycoside is used.

15. A method of promoting CES1 expression, wherein,

sulforaphane, or a plant or extract thereof containing sulforaphane or its glycoside is used.

16. The method of claim 15 or 16,

the method is used by a person using a composition containing parabens.

Technical Field

The present invention relates to a method for evaluating or selecting a sensory stimulation reducing agent for suppressing sensory stimulation due to parabens, and a sensory stimulation reducing agent for alleviating the sensory stimulation due to parabens.

Background

In skin external preparations such as skin cosmetics, preservatives are often mixed in the preparations in order to prevent bacterial contamination and proliferation. Among them, parabens are most often mixed in skin external preparations as preservatives having high antibacterial activity and safety. However, people who are allergic to ultraviolet rays, dryness, chemicals, and the like, that is, people who are so-called sensitive to skin, sometimes feel unpleasant and temporary stinging sensations such as stinging sensations and hot and spicy sensations due to parabens.

On the other hand, in the prior art, as an agent for suppressing the skin irritation, for example, the following are reported: a irritation inhibitor composed of a polysaccharide derivative having a hydrophobic group for inhibiting irritation caused by a surfactant (patent document 1), and a irritation inhibitor for hair dye for inhibiting irritation caused by a hair dye (patent document 2). In addition, as a paraben-based irritation inhibitor, an inhibitor that can use TRPA1 has been reported (patent document 3).

On the other hand, it is known that sprouted broccoli, which is a sprout of broccoli (Brassica oleracea. italica) belonging to Brassica of brassicaceae, is a plant having a high sulforaphane content, and its extract has been reported to have a preventive and improving effect on whitening, wrinkles, sagging, and the like (patent document 4), a sebum production promoting effect (patent document 5), and the like.

However, there has been no report on the effect of preventing and improving sensitive muscles of an extract of germinated broccoli or sulforaphane.

[ patent document 1] Japanese patent application laid-open No. 2001-64185

[ patent document 2] Japanese patent laid-open No. 2002-363053

[ patent document 3] Japanese patent laid-open No. 2008-79528

[ patent document 4] Japanese patent laid-open No. 2003-155221

[ patent document 5] Japanese patent application No. 2009-114152

Disclosure of Invention

The present invention provides the following 1) to 9).

1) A method for evaluating or selecting a sensory stimulation reducing agent for reducing sensory stimulation due to parabens, which comprises the following steps (1) to (3):

(1) a step of contacting a test substance with a cell capable of expressing CES 1;

(2) measuring the expression of CES1 in the cell; and

(3) and (3) evaluating a test substance that promotes the expression of CES1 as a sensory stimulation reducing agent that reduces sensory stimulation by parabens based on the result of the measurement in (2).

2) A sensory stimulation reducing agent contains sulforaphane, or a plant or an extract thereof containing sulforaphane or a glycoside thereof as an active ingredient.

3) CES1 expression promoter contains sulforaphane, or plant or extract containing sulforaphane or its glucoside as effective component.

4) Use of sulforaphane, or a plant or an extract thereof containing sulforaphane or a glycoside thereof for the manufacture of a sensory stimulation reducing agent.

5) Use of sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof for the manufacture of a CES1 expression promoter.

6) Sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof, for reducing sensory stimulation.

7) Sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof, for promoting expression of CES 1.

8) Non-therapeutic use of sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof, for reducing sensory stimulation.

9) Non-therapeutic use of sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof, for promoting expression of CES 1.

10) A method for reducing sensory stimulation comprises using sulforaphane, or a plant or extract thereof containing sulforaphane or its glycoside.

11) A method for promoting CES1 expression comprises using sulforaphane, or plant or extract containing sulforaphane or its glycoside.

Drawings

FIG. 1 shows the amounts of I L-1 α produced in the three-dimensional cultured epidermis, MP, added methylparaben, PHBA, added parahydroxybenzoic acid, and the data are the mean. + -. SD.

Fig. 2 is a skin sensory stimulation score for a person with sensitive skin. MP: smearing methylparaben, PHBA: smearing p-hydroxybenzoic acid.

Fig. 3 shows CES1 expression levels of healthy subjects and sensitive skin subjects. Data are mean ± SD.

FIG. 4 shows the metabolic activity of methylparaben in healthy and sensitive skin subjects. Data are mean ± SD.

FIG. 5 shows the CES1 expression promoting effect of the sprouted broccoli extract.

FIG. 6 is a graph showing the effect of the extract of germinated broccoli on reduction of sensory stimulation due to methylparaben.

Detailed Description

The present invention relates to a method for evaluating or selecting a sensory stimulation reducing agent for reducing sensory stimulation due to parabens. The present invention also relates to a sensory stimulation reducing agent for reducing sensory stimulation due to parabens.

The present inventors have studied sensory stimulation caused by parabens and, as a result, have found that: methylparaben is metabolized in vivo by the metabolic enzyme carboxylesterase 1(CES1) and converted to Parabens (PHBA), where there is little sensory stimulation; further, expression of CES1 is reduced in people with sensitive muscle compared to healthy people; and found that: by using CES1 expression as an index, a sensory stimulation reducing agent that reduces sensory stimulation by parabens can be searched for or evaluated. And, found that: sulforaphane or an extract of a sulforaphane-containing plant such as a sprouted broccoli has an expression promoting effect of CES1, and can be a sensory stimulation reducing agent having an effect of reducing sensory stimulation due to parabens.

According to the present invention, a novel sensory stimulation reducing agent which rapidly decomposes parabens in vivo and reduces sensory stimulation caused by parabens can be searched for or evaluated.

Further, according to the present invention, a novel sensory stimulation reducing agent which rapidly decomposes parabens in a living body and reduces sensory stimulation caused by the parabens can be provided.

In the present invention, examples of the "parabens" include: parabens such as methyl paraben (p-hydroxybenzoic acid methyl ester), ethyl paraben (p-hydroxybenzoic acid ethyl ester), propyl paraben (p-hydroxybenzoic acid propyl ester), isopropyl paraben (p-hydroxybenzoic acid isopropyl ester), butyl paraben (p-hydroxybenzoic acid butyl ester), isobutyl paraben (p-hydroxybenzoic acid isobutyl ester), and benzyl paraben (p-hydroxybenzoic acid benzyl ester), and sodium salts thereof.

The term "reducing sensory irritation due to parabens" means suppressing or alleviating a temporary unpleasant sensation of irritation such as skin irritation or hot and spicy sensation due to parabens.

In the present invention, "CES 1" refers to carboxylesterase 1, and is also known as serine esterase 1 or SES 1. The enzyme is a member of the family of mammalian liver carboxylesterases (EC 3.1.1.1) and hydrolyses endogenous substrates having ester, thioester or amide functional groups. In humans, expression in lung and skin is reported in addition to liver, and is involved in metabolic activation of drugs and detoxification of foreign substances in vitro such as chemical substances.

As shown in the following examples, Methyl Paraben (MP) or p-hydroxybenzoic acid (PHBA) was added to a human three-dimensional cultured epidermal specimen, and the amount of inflammatory cytokine (I L-1 α) in the culture medium was measured, and as a result, I L-1 α was increased in the case of MP addition, but almost no I L-1 α was observed in the case of PHBA addition (FIG. 1). in addition, MP or PHBA was applied to the skin of a person who recognized sensitive muscles, and the irritation thereof was tested, and as a result, some irritation was felt in the case of MP application, while almost no irritation was felt in the case of PHBA application (FIG. 2).

Further, the skin was collected from healthy subjects and sensitive skin subjects, and the metabolic activity of MP and the expression level of CES1 in the skin were measured, and as a result, it was confirmed that: compared with the skin of a healthy person, the metabolic activity of MP in the skin of a sensitive skin person is low, and the CES1 expression level is very low (fig. 3 and 4).

These results suggest that: a substance that promotes the expression of CES1 is useful as a sensory stimulation reducing agent that reduces sensory stimulation by parabens, and a sensory stimulation reducing agent that reduces sensory stimulation by parabens can be evaluated or selected using the expression of CES1 as an index.

A method for evaluating or selecting a sensory stimulation reducing agent for reducing sensory stimulation due to parabens, which comprises the following steps (1) to (3):

(1) a step of contacting a test substance with a cell capable of expressing CES 1;

(2) measuring the expression of CES1 in the cell; and

(3) and (3) evaluating a test substance that promotes the expression of CES1 as a sensory stimulation reducing agent that reduces sensory stimulation by parabens based on the result of the measurement in (2).

Here, examples of the "cell capable of expressing CES 1" include: a cell having a CES1 gene and having the ability to express the gene, and a cell having a CES1 gene introduced thereto so as to be able to express the gene. The cells may be cells collected from a living body, cells contained in a tissue or an organ collected from a living body, or cultured cells. The cell is preferably a cell derived from a mammal, more preferably a cell derived from a human.

Examples of the cells having the CES1 gene and the ability to express the gene include: the cells derived from all tissues of a living body preferably include: examples of skin-derived cells collected from mammals (preferably humans) include cells derived from epidermal tissues (epidermal keratinocytes and the like) and cells derived from dermal tissues (fibroblasts and the like), and cell cultures, organ cultures and the like derived from these cells are preferably cells derived from epidermal tissues, and more preferably epidermal keratinocytes.

A foreign cell capable of expressing the CES1 gene can be obtained by introducing an expression vector containing the CES1 gene into an arbitrary mammalian cell and transforming the cell. The transcription regulatory region (promoter or the like) in the expression vector is not particularly limited as long as it can control expression in vivo, and it is preferable to use the transcription regulatory region (promoter or the like) of the CES1 gene. In addition, as a cell capable of expressing the CES1 gene, a cell in which a vector to which a reporter gene (reporter gene) is linked is introduced into a transcription regulatory region (e.g., a promoter) of the CES1 gene may be used. Methods for producing a vector incorporating a transcription regulatory region (promoter or the like) of the CES1 gene or CES1 gene, and methods for introducing a vector into mammalian cells are known to those skilled in the art.

In addition to the cultured cells derived from the above-mentioned naturally occurring cells having the CES1 gene and capable of expressing the gene and the exogenous cells into which the CES1 gene has been introduced so as to be expressible, cultured cells derived from a strain capable of expressing CES1, preferably cultured cells derived from the skin collected from a mammal (preferably a human being), more preferably cultured epidermal keratinocytes derived from a normal human being, may be mentioned as the cultured cells.

The test substance to be contacted with the above-mentioned cells is not particularly limited as long as it is desired to be used as a sensory stimulation reducing agent for reducing sensory stimulation by parabens, and examples thereof include: animal and plant, marine organism, microorganism, etc. and their extracts; natural ingredients derived from these; synthesizing a compound; and mixtures and combinations thereof.

The method of contacting the test substance with the above-mentioned cells may be any method known in the art, and examples thereof include: the test substance is added to the cell culture medium, or directly to the cells (e.g., dropped, smeared, spread, sprayed, patch, etc.). The concentration and contact amount of the test substance may be appropriately set based on the form, chemical property, cytotoxicity, intensity of expected skin sensitization, and the like of the test substance. For example, there may be mentioned: a predetermined amount of the test substance diluted to an appropriate concentration was subjected to a reaction at 37 ℃ with 5% CO₂Is exposed to the cells for 24 to 48 hours.

The expression of CES1 in the above cells can be measured by using, as indicators, the expression of CES1 protein, the activity of CES1 protein, the expression of CES1mRNA encoding the protein, the activation of the CES1 gene promoter, and the like. The measurement method of the parameters (for example, protein expression, biological activity of protein, mRNA (Messenger RNA) expression, activation of CES1 gene promoter, etc.) to be used as the index may be performed according to a method known in the art.

Examples of the measurement method include, for example, the measurement method of mRNA expression: dot blot (dot blot) method, Northern blot (Northern blot) method, RT-PCR (Reverse Transcription-Polymerase chain reaction), real-time RT-PCR, microarray, etc., and combinations thereof.

Examples of a method for measuring the activity of the CES1 gene promoter include: fluorescence-optical measurement of promoter activity or transcriptional activity of a reporter gene (reporter assay) was used.

Examples of The method for measuring protein expression include SDS-PAGE (Sodium dodecyl sulfate-Polyacrylamide gel electrophoresis), chromatography, immunological measurement methods (e.g., immunohistochemistry, E L ISA (enzyme-linked immunosorbent assay), Western blotting (Western Blot), immunoprecipitation, colorimetric quantification, mass analysis, and The like, and combinations thereof.

The activity of CES1 protein can be measured, for example, by exposing methylparaben, which is a substrate of CES1, to CES 1-expressing cells, culturing for a certain period of time, and then quantifying the amount of paraben in the culture broth using HP L C.

Then, based on the expression measured as described above, the expression promoting effect of CES1 on the test substance is evaluated, for example, as follows: the comparison is performed before and after the addition of the test substance, or the comparison is performed between the test substance-added group and the test substance-non-added group or the control substance-added group. Alternatively, the evaluation may be performed by comparing the measurement results between test substances at various concentrations.

The substance to be tested, which has enhanced or promoted the expression of CES1, is selected as a sensory stimulation reducing agent for reducing sensory stimulation due to parabens.

The sensory stimulation reducing agent obtained in this manner, which reduces sensory stimulation due to parabens, can be used by being mixed with cosmetics, pharmaceuticals, and the like containing parabens, because it reduces sensory stimulation due to parabens.

By the above method of the present invention, it was found that: sulforaphane, and extract of sulforaphane-containing plant such as germinated broccoli have expression promoting effect of CES1, and can be used as sensory stimulation reducing agent.

That is, the present invention further provides a sensory stimulation reducing agent or a CES1 expression promoter containing sulforaphane, or a plant or an extract thereof containing sulforaphane or a glycoside thereof as an active ingredient.

Here, Sulforaphane (Sulforaphane) refers to 4- (methylsulfinyl) butyl isothiocyanate.

Sulforaphane can be obtained by separating and purifying a plant containing sulforaphane or a glycoside thereof by a known method. Further, sulforaphane, which is isolated and purified, is commercially available from Sigma-Aldrich company and the like, and this commercially available product can be used in the present invention.

In the present invention, not only the isolated sulforaphane can be an active ingredient of a sensory stimulation reducing agent or a CES1 expression promoter, but also a plant or an extract thereof containing the sulforaphane or its glycoside can be an active ingredient of a sensory stimulation reducing agent or a CES1 expression promoter.

Examples of the plant containing sulforaphane or a glycoside thereof include: cruciferous (Brassicaceae) plants, preferably Brassica (Brassica) plants. Specifically, there may be mentioned: broccoli, cauliflower, cabbage, and the like. Among them, broccoli (Brassica oleracea) is preferable in terms of the content of sulforaphane, and a germinated broccoli (also referred to as "broccoli embryo") before it becomes an adult after germination is more preferable.

Examples of the glycoside of sulforaphane include: sulforaphane glucosinolate, and the like.

As the plant containing sulforaphane or its glycoside, a pulverized product, freeze-dried product, squeezed juice or the like of all or a part of the plant can be used. For example, in the case of a sprouted broccoli, it is preferable to use a dry ground product of a part of or the whole of a bud, cotyledon, seed, or root of a sprouted body.

Examples of the extract of the above plant include: various solvent extracts, dilutions thereof, concentrates thereof or dried powders thereof obtained by extraction using a known extraction method. Known extraction methods include, for example: dipping, extracting, leaching, solid-liquid extracting, reflux extracting, supercritical extracting, ultrasonic extracting, microwave extracting and the like. For example, as for impregnation, there can be mentioned: dipping and leaching for 1 hour to 4 weeks at the temperature of 0 ℃ to the boiling point of the solvent (preferably 15 to 40 ℃). As for the solid-liquid extraction, there can be mentioned: stirring or shaking is carried out at 0 ℃ to the boiling point of the solvent (preferably 15 to 40 ℃) for 30 minutes to 2 weeks at 30 to 1000 rpm. In addition, in order to prevent oxidation of the extract, the following methods may be used in combination: boiling for degassing or introducing an inert gas such as nitrogen gas to remove dissolved oxygen, i.e., extracting under a so-called non-oxidizing atmosphere. In addition, in the case of reflux extraction, extraction can be performed using an extraction instrument such as a soxhlet extractor.

As the solvent used for extraction, either a polar solvent or a nonpolar solvent may be used. Specific examples of the solvent include: water; alcohols such as monohydric alcohols, dihydric alcohols, and polyhydric alcohols; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; linear or cyclic ethers such as diethyl ether and tetrahydrofuran; polyethers such as polyethylene glycol; saturated or unsaturated hydrocarbons such as hexane; aromatic hydrocarbons such as benzene and toluene; halogenated hydrocarbons such as dichloromethane, chloroform, dichloroethane, carbon tetrachloride and the like; pyridines; dimethyl sulfoxide; acetonitrile; carbon dioxide, supercritical carbon dioxide; fats, waxes, other oils; and mixtures of these. From the viewpoint of pharmacological activity and versatility, preferable are: water, alcohols, and mixtures thereof.

The alcohols are not particularly limited, and examples thereof include: monohydric alcohols such as methanol, ethanol, propanol, butanol, pentanol, hexanol, heptanol, and octanol; diols such as 1, 3-butanediol, ethylene glycol, propylene glycol, 1, 4-butanediol, 1, 5-pentanediol, and 1, 6-hexanediol; and trihydric or higher alcohols such as glycerin. Among them, monohydric alcohols and dihydric alcohols are preferable from the viewpoint of pharmacological activity and operability. The alcohol is preferably an alcohol having 1 to 10 carbon atoms, and more preferably an alcohol having 1 to 4 carbon atoms.

Preferred examples of the alcohols include: methanol, ethanol, 1, 3-butanediol, n-propanol, isopropanol, n-butanol, isobutanol, primary butanol, tertiary butanol, and the like.

The concentration of the alcohol in the aqueous solution of the alcohol is preferably 20% by mass or more, more preferably 30% by mass or more, more preferably 40% by mass or more, and preferably 80% by mass or less, more preferably 70% by mass or less, and further preferably 60% by mass or less. The concentration of the alcohol in the aqueous solution of the alcohol is preferably 20 to 80% by mass, more preferably 30 to 70% by mass, and still more preferably 40 to 60% by mass.

Among the above alcohols, ethanol is more preferable from the viewpoint of general versatility. Therefore, as more preferable solvents for preparing the extract of the above plant, there can be mentioned: water, ethanol, and ethanol aqueous solution.

The amount of solvent used in the extraction treatment is preferably 1 to 1000m L per 1g of plant (in terms of dry mass), and the volume is herein referred to as 25 ℃ volume, and the dry mass is referred to as the mass of the dry solid matter of the remaining components after drying the sample in an electric constant temperature dryer at 105 ℃ for 3 hours to remove volatile matter, and the extraction conditions are not particularly limited as long as sufficient extraction treatment can be performed, and for example, the extraction time is preferably 1 hour or more, more preferably 3 days or more, more preferably 1 week or more, on the other hand, preferably 2 months or less, more preferably 5 weeks or less, and more preferably 2 weeks or less, and the extraction temperature is preferably 0 ℃ or more, more preferably 5 ℃ or more, on the other hand, preferably a solvent boiling point or less, more preferably 90 ℃ or less, and generally, if low temperature, extraction is performed for a long time, and if high temperature, extraction is performed for a short time, and the extraction conditions are exemplified by those skilled in the art, but are not limited appropriately.

In the present invention, the plant extract may be used as it is, or the extract may be diluted, concentrated or dried to prepare a powder or a paste. When the extract is used after freeze-drying, it may be diluted with a solvent generally used for extraction, for example, a solvent such as water, ethanol, propylene glycol, 1, 3-butanediol, a water-ethanol mixture, a water-propylene glycol mixture, or a mixture of water and 1, 3-butanediol. Further, the compound may be used by being encapsulated in vesicles such as liposomes or microcapsules.

The plant extract thus obtained may be subjected to separation/purification steps such as treatment with a synthetic adsorbent, filtration treatment, and concentration treatment, if necessary, to increase the content of sulforaphane. As the extract of the sprouted broccoli having the increased sulforaphane content, a "broccoli embryo extract" (ORYZA OIL CROSS) is sold, and in the present invention, the commercially available product can be used.

As shown in the following examples, sulforaphane and an extract of germinated broccoli, which is a sulforaphane-containing plant, have an effect of promoting the expression of CES1 and an effect of reducing sensory stimulation due to parabens.

Therefore, sulforaphane, a plant containing sulforaphane or a glycoside thereof, or an extract thereof can be used as a sensory stimulation reducing agent or a CES1 expression promoter, and can be used for the purpose of reducing sensory stimulation, the purpose of promoting the expression of CES1, and the purpose of producing a sensory stimulation reducing agent or a CES1 expression promoter.

Here, "use" may be administration or ingestion to a human or non-human animal, and may be therapeutic use or non-therapeutic use. The term "non-therapeutic" means a concept not including medical practice, that is, a concept not including a method of performing surgery, treatment or diagnosis on a human, and more specifically, a concept not including a method of performing surgery, treatment or diagnosis on a human by a doctor or a person who receives an instruction from the doctor.

The "reduction in sensory stimulation" in the "sensory stimulation reducing agent" of the present invention means to suppress or alleviate an unpleasant sensation of temporal irritation such as a skin prickling sensation or a hot and spicy sensation caused by parabens. Here, "parabens" means those as mentioned above.

The sensory stimulation reduction effect can be measured by the sensory evaluation shown in the examples described below.

The "CES 1 expression" in the "CES 1 expression promoter" includes the expression of the CES1 protein, the activity of the CES1 protein, the expression of CES1mRNA encoding the protein, and the activity of a CES1 gene promoter as described above, and preferably, the CES1mRNA expression.

The sensory stimulation reducing agent and the CES1 expression promoter may be a preparation using sulforaphane, a plant containing sulforaphane or a glycoside thereof, or an extract thereof, alone, or a composition combined with an oil, a pigment, a flavor, a preservative, a chelating agent, a pigment, an antioxidant, a vitamin, a mineral, a sweetener, a flavoring agent, a preservative, a binder, a bulking agent, a disintegrating agent, a surfactant, a lubricant, a dispersing agent, a buffer, a coating agent, a carrier, a diluent, or an additive or excipient used in various preparations such as a pharmaceutical preparation, a cosmetic preparation, a quasi-drug preparation, and a daily use product. The form of the polymer is not particularly limited, and the polymer may be prepared in any form such as a solution, emulsion, suspension, gel, solid, powder, granule, or aerosol.

The amount of sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof in the composition is 0.0001% by mass or more, preferably 0.001% by mass or more, more preferably 0.01% by mass or more, further preferably 0.1% by mass or more, and 99% by mass or less, preferably 95% by mass or less, more preferably 90% by mass or less, further preferably 80% by mass or less of the total mass of the preparation, based on the sulforaphane. The amount of the organic solvent is 0.001 to 99% by mass, preferably 0.01 to 95% by mass, more preferably 0.1 to 90% by mass, and still more preferably 1 to 80% by mass. In addition, when the composition contains "a plant or an extract thereof containing sulforaphane or a glycoside thereof", the content thereof is 0.001 mass% or more, preferably 0.01 mass% or more, more preferably 0.1 mass% or more, and further preferably 1 mass% or more, and 99 mass% or less, preferably 95 mass% or less, more preferably 90 mass% or less, and further preferably 80 mass% or less of the total mass of the preparation in terms of dry solid matter components. The amount of the organic solvent is 0.001 to 99% by mass, preferably 0.01 to 95% by mass, more preferably 0.1 to 90% by mass, and still more preferably 1 to 80% by mass.

The sensory stimulation reducing agent or CES1 expression promoter of the present invention can be used by, for example, mixing it with the above composition containing a paraben (e.g., a skin cleanser, a shampoo, a makeup preparation, a body wash, a permanent wave preparation, a hair dye, a soap, a kitchen detergent, a laundry detergent, a cosmetic such as a toothpaste, a quasi-drug, a pharmaceutical, a daily product, etc.), or preparing it as a composition different from the composition containing a paraben, and can reduce the stimulation caused by the paraben by using the composition simultaneously with or before or after the use of the composition. That is, the target to which the sensory stimulation reducing agent or the CES1 expression promoter of the present invention is applied is preferably: a human being who uses a composition containing parabens and is required to have a reduced irritation caused by the parabens.

When the sensory stimulation reducing agent and the CES1 expression promoter of the present invention are used together with a composition containing parabens, the amount of the sensory stimulation reducing agent and the CES1 expression promoter used is not particularly limited as long as it can reduce sensory stimulation, and for example, the sulforaphane of the present invention is used in an amount of 1 part by mass of parabensOr a plant or extract thereof containing sulforaphane or a glycoside thereof, in terms of sulforaphane, in the following ratio: preferably 10^-7At least, more preferably 10 parts by mass^-6At least, preferably 10 parts by mass^-5The amount is preferably not less than 10 parts by mass, more preferably not more than 1 part by mass, and still more preferably not more than 0.1 part by mass. In addition, the following ratio can be set: preferably 10^-7About 10 parts by mass, more preferably about 10 parts by mass^-6About 1 part by mass, more preferably about 10^-50.1 part by mass.

The present invention further discloses the following embodiments with respect to the above embodiments.

<1> a method for evaluating or selecting a sensory stimulation reducing agent for reducing sensory stimulation due to parabens, which comprises the following steps (1) to (3):

(1) a step of contacting a test substance with a cell capable of expressing CES 1;

(2) measuring the expression of CES1 in the cell; and

(3) and (3) evaluating a test substance that promotes the expression of CES1 as a sensory stimulation reducing agent that reduces sensory stimulation by parabens based on the result of the measurement in (2).

<2> the method according to <1>, wherein the parabens are at least one selected from the group consisting of methyl paraben, sodium methyl paraben, ethyl paraben, butyl paraben, propyl paraben, isobutyl paraben, isopropyl paraben and benzyl paraben.

<3> the method <2>, wherein the paraben is methylparaben.

<4> the method of any one of <1> to <3>, wherein the cells capable of expressing CES1 are cells derived from human epidermal tissue, preferably normal human cultured epidermal keratinocytes.

<5> the method according to any one of <1> to <4>, wherein the expression of CES1 is measured using at least one selected from the group consisting of expression of CES1 protein, activity of CES1 protein, expression of CES1mRNA encoding the protein, and activity of a promoter of CES1 gene as an indicator, preferably expression of CES1mRNA as an indicator.

<6> a sensory stimulation reducing agent comprising sulforaphane, or a plant or an extract thereof containing sulforaphane or a glycoside thereof as an active ingredient.

<7> a CES1 expression promoter comprising sulforaphane, a plant containing sulforaphane or a glycoside thereof, or an extract thereof as an active ingredient.

<8> use of sulforaphane, or a plant or an extract thereof containing sulforaphane or a glycoside thereof for the manufacture of a sensory stimulation reducing agent.

<9> use of sulforaphane, or a plant or an extract thereof containing sulforaphane or a glycoside thereof for the manufacture of a CES1 expression promoter.

<10> a sulforaphane, or a plant or an extract thereof containing sulforaphane or its glycoside, for reducing sensory stimulation.

<11> a sulforaphane, or a plant or an extract thereof containing sulforaphane or its glycoside, for promoting expression of CES 1.

<12> non-therapeutic use of sulforaphane, or a plant or extract thereof containing sulforaphane or its glycosides for reducing sensory stimuli.

<13> non-therapeutic use of sulforaphane, or a plant or extract thereof containing sulforaphane or a glycoside thereof, for promoting expression of CES 1.

<14> a method for reducing sensory stimulation, wherein sulforaphane, or a plant or an extract thereof containing sulforaphane or its glycoside is used.

<15> a method for promoting CES1 expression, which comprises using sulforaphane, or a plant or extract thereof containing sulforaphane or its glycoside.

<16> the method as stated in <14> or <15>, wherein the method is applied to a human using a composition containing parabens.

<17> in <6>, <8>, <10>, <12> or <14> above, the sensory stimulation is a stimulation sensation caused by parabens.

<18> in the above <6> to <17>, the plant containing sulforaphane or its glycoside is a sprouted broccoli.

<19> in the above <6> to <18>, sulforaphane, or a plant containing sulforaphane or its glycoside or an extract thereof is mixed with a composition containing parabens and used, or a composition different from the composition is prepared and used simultaneously with or before and after the composition is used.

<20>In the above<19>In (1), sulforaphane, or a plant or an extract thereof containing sulforaphane or its glycoside is used in the following ratio: the amount of sulforaphane, or a plant or extract thereof containing sulforaphane or its glycoside is preferably 10 parts by mass based on sulforaphane, based on 1 part by mass of parabens^-7At least, more preferably 10 parts by mass^-6At least, preferably 10 parts by mass^-5At least, and preferably 10 parts by mass or less, more preferably 1 part by mass or less, further preferably 0.1 part by mass or less, or preferably 10 parts by mass or less^-7About 10 parts by mass, more preferably about 10 parts by mass^-6About 1 part by mass, more preferably about 10^-50.1 part by mass.

<21> in the above <6> to <9>, the amount of sulforaphane, or a plant containing sulforaphane or its glycoside or an extract thereof to be mixed in the composition is as follows: the content of sulforaphane is 0.0001% by mass or more, preferably 0.001% by mass or more, more preferably 0.01% by mass or more, and further preferably 0.1% by mass or more, and 99% by mass or less, preferably 95% by mass or less, more preferably 90% by mass or less, and further preferably 80% by mass or less, or 0.001 to 99% by mass, preferably 0.01 to 95% by mass, more preferably 0.1 to 90% by mass, and further preferably 1 to 80% by mass, based on the total mass of the preparation.

<22> in the above <6> to <9>, the mixing amount of the plant or the extract thereof containing sulforaphane or its glycoside in the composition is as follows: the content of the active ingredient is 0.001 mass% or more, preferably 0.01 mass% or more, more preferably 0.1 mass% or more, and further preferably 1 mass% or more, and 99 mass% or less, preferably 95 mass% or less, more preferably 90 mass% or less, and further preferably 80 mass% or less, or 0.001 to 99 mass%, preferably 0.01 to 95 mass%, more preferably 0.1 to 90 mass%, and further preferably 1 to 80 mass% of the total mass of the preparation in terms of the dry solid content.