阅读说明：本技术 植物甾酮的新型用途 (Novel use of phytosterol ) 是由 长岛直 井上奈穗 于 2019-08-29 设计创作，主要内容包括：根据本发明,提供含有植物甾酮的用于减少皮下脂肪的经皮吸收剂。由此,进一步提供皮下脂肪的减少方法。(According to the present invention, there is provided a percutaneous absorption agent for reducing subcutaneous fat, which contains a phytosterol. Thus, a method of reducing subcutaneous fat is further provided.)

1. A percutaneous absorption agent for reducing subcutaneous fat, which contains phytosterol.

2. The percutaneous absorption agent as claimed in claim 1, wherein the phytosterol is a composition containing the phytosterol.

3. The percutaneous absorption agent as claimed in claim 2, wherein the composition containing phytosterol comprises 5-en-3-one body of phytosterol, 4-en-3-one body of phytosterol and 4-en-3, 6-dione body of phytosterol.

4. The percutaneous absorption agent as claimed in claim 1, wherein the phytosterol is campest-5-en-3-one.

5. The percutaneous absorption agent as claimed in any one of claims 1 to 4, which is a cosmetic.

6. A transdermal absorbent according to any one of claims 1 to 4 which is a drug.

7. A phytosterol for use in reducing subcutaneous fat, which is applied in a manner suitable for use on the skin.

8. A method of reducing subcutaneous fat in a subject, comprising: the phytosterols are applied to the skin of a subject in need of a reduction of subcutaneous fat.

Technical Field

The present invention relates to a novel use of phytosterols.

Background

The excessive accumulation of fat is believed to be caused by various causes such as insufficient energy consumption due to excessive intake of nutrients, insufficient exercise, etc., abnormal stress and hormone balance, and sequelae of diseases. The accumulation site of fat includes organs such as liver, intestinal membrane tissue, subcutaneous tissue, etc. Especially, subcutaneous tissue occupies a large area as a site for accumulation of fat. Among them, a large amount of fat is likely to accumulate in the regions from the upper arm, abdomen, and waist to the thigh, and this is also considered a problem from the viewpoint of appearance.

Many attempts have been made to reduce excessive accumulation of subcutaneous fat, including treatment and prevention in medical care, and prevention of diet and various beverages by daily intake. For example, food materials intended to regulate the metabolism of ingested egg lipids, promote the combustion of body fat, or inhibit the accumulation of body fat by food materials such as L-carnitine or meat peptides containing the same, algae extracts, carotenoids, anthocyanins, green tea catechins, and the like have been proposed.

However, the present situation is that the number of materials effective for preventing and improving obesity symptoms is small, and materials that are sufficiently satisfactory are hardly found. Therefore, materials that inhibit the absorption of fat in the diet, inhibit the accumulation of fat in the body, promote the combustion of body fat, or promote the metabolism of body fat are required.

For phytosterol (phytosterol, Japanese: フィトステロール, phytosterone) as a fermentation product of phytosterol (phytosterol, Japanese: フィトステノン, phytosterone), there have been reported: when added to a high-fat diet and fed, visceral fat and subcutaneous fat are reduced (for example, non-patent documents 1 to 3). It can be considered that: the reduction effect of visceral fat and subcutaneous fat by oral administration of phytosterol is achieved by various mechanisms of action including enhancement of fat metabolism by β oxidation, inhibition of fatty acid synthesis, and in vitro excretion of lipids (for example, non-patent documents 1 and 2). However, it is not known that phytosterols can be applied to the skin of a subject to reduce the subcutaneous fat in the subject.

Here, phytosterol has been proposed as an additive for foods and beverages, drugs, and the like (for example, patent documents 1 to 3). Further, a method for producing 5-en-3-one (a type of phytosterol) of sterol in a good yield has been proposed (for example, patent document 4).

Documents of the prior art

Patent document

Patent document 1: japanese patent laid-open No. 2001-240544

Patent document 2: japanese patent laid-open publication No. 2003-48837

Patent document 3: japanese patent laid-open publication No. 2011-51939

Patent document 4: japanese laid-open patent publication No. 2007-284348

Non-patent document

Non-patent document 1: suzuki Pongfu et al, Nippon Rinsho (2003),61, suppl.6, p.698-703

Non-patent document 2: kunio Suzuki, et al, J.Nutr.Sci.Vitaminol. (2007),53, p.63-67

Non-patent document 3: obesity study (2007),13(3), p.244-249

Disclosure of Invention

Problems to be solved by the invention

Under the above circumstances, a method of reducing subcutaneous fat and the like are further required.

Means for solving the problems

The present inventors have conducted intensive studies and, as a result, have found that: the present invention has been completed by applying phytosterol to the skin of a subject, thereby reducing the subcutaneous fat and the like of the subject.

Namely, the following percutaneous absorbents and the like are provided.

[1-1] A percutaneous absorption agent for reducing subcutaneous fat, which contains a phytosterol.

[1-2] the percutaneous absorption agent according to the above [1-1], wherein the phytosterol is a composition containing the phytosterol.

[1-3] the percutaneous absorption agent as described in the above [1-2], wherein the composition containing a phytosterol comprises a 5-en-3-one body of a phytosterol, a 4-en-3-one body of a phytosterol and a 4-en-3, 6-dione body of a phytosterol.

[1-4] the percutaneous absorption agent according to the above [1-1], wherein the phytosterol is campest-5-en-3-one.

[1-5] the percutaneous absorbent according to any one of [1-1] to [1-4], which is a cosmetic.

[1-6] the percutaneous absorption agent according to any one of [1-1] to [1-4], which is a drug.

[2-1] A phytosterol for reducing subcutaneous fat, which is used in a manner suitable for skin.

[2-2] the phytosterol according to the above [2-1], wherein the phytosterol is a composition containing the phytosterol.

[2-3] the phytosterol according to the above [2-3], wherein the composition containing phytosterol comprises 5-en-3-one body of phytosterol, 4-en-3-one body of phytosterol and 4-en-3, 6-dione body of phytosterol.

[2-4] the phytosterol according to the above [2-1], wherein the phytosterol is campest-5-en-3-one.

[2-5] the phytosterol according to any one of the above [2-1] to [2-4], which is used as a cosmetic.

[2-6] the phytosterol according to any one of the above [2-1] to [2-4], for use as a medicament.

[3-1] A method of reducing subcutaneous fat in a subject, comprising applying phytosterol to the skin of a subject in need of subcutaneous fat reduction.

[3-2] the method according to the above [3-1], wherein the phytosterol is a composition containing the phytosterol.

[3-3] the method according to the above [3-2], wherein the composition containing phytosterol comprises 5-en-3-one body of phytosterol, 4-en-3-one body of phytosterol and 4-en-3, 6-dione body of phytosterol.

[3-4] the method according to the above [3-1], wherein the phytosterol is campest-5-en-3-one.

[3-5] the method according to any one of [3-1] to [3-4], which is a cosmetic method.

[3-6] the method according to any one of [3-1] to [3-4], which is a therapeutic method.

ADVANTAGEOUS EFFECTS OF INVENTION

According to the present invention, there is further provided a method of reducing subcutaneous fat.

The transdermal absorbent of a preferred embodiment can reduce subcutaneous fat in a subject by being applied to the skin of the subject. Another preferred embodiment of the transdermal absorption agent is applied to the skin of a subject to reduce the metabolism of fat excessively accumulated under the skin of the subject. In still another preferred embodiment, the transdermal absorption agent can reduce fat which is locally or widely excessively accumulated under the skin of a subject and which has a problem from the viewpoint of health or beauty, and can improve the health or beauty of the subject.

Drawings

Fig. 1 is a graph showing the effect of phytosterols on subcutaneous fat. E: ethanol solution (control), C: campest-5-en-3-one, P: a composition contains phytosterol.

Fig. 2 is a graph showing the effect of phytosterols on mesenteric fat. E: ethanol solution (control), C: campest-5-en-3-one, P: a composition contains phytosterol.

Fig. 3 is a graph showing the effect of phytosterols on peri-spermatic fat. E: ethanol solution (control), C: campest-5-en-3-one, P: a composition contains phytosterol.

Fig. 4 is a graph showing the effect of phytosterol on perirenal fat. E: ethanol solution (control), C: campest-5-en-3-one, P: a composition contains phytosterol.

Fig. 5 is a graph showing the effect of phytosterol on the liver. E: ethanol solution (control), C: campest-5-en-3-one, P: a composition contains phytosterol.

Detailed Description

The present invention will be described in detail below.

Some embodiments provide a transdermal absorbent for reducing subcutaneous fat, which contains a phytosterol. The phytosterol may be used alone in 1 kind, or in combination of two or more kinds.

"phytosterol" is an alkenone compound having a structure in which the hydroxyl group on the carbon at position 3 of phytosterol is substituted with an oxo group, that is, having a carbonyl group at position 3. In the present specification, "phytosterol" is sometimes referred to as "phytosterol, japanese: plant ステノン "or" 3-ketone bodies of phytosterols ", and" phytosterols "are referred to as" phytosterols, japanese: plant ステロール ".

In the present specification, the term "phytosterol" is used in a meaning that includes phytosterols and its analogues. In addition, in the present specification, the term "phytosterol" is used in a meaning including phytosterol and its analogues.

The structure of the phytosterol is not particularly limited as long as it has a carbonyl group at the 3-position of the sterol skeleton. The phytosterols may also have other substituents. The phytosterol may also have more than 1 carbonyl group at positions other than the 3-position of the sterol backbone. Examples of the phytosterol having a carbonyl group at a position other than the 3-position include a 3, 6-diketone of phytosterol (having carbonyl groups at the 3-position and the 6-position of the sterol skeleton). The phytosterol may have a functional group such as an alkyl group (e.g., methyl group and ethyl group) or a hydroxyl group on the sterol skeleton or a side chain thereof. In addition, the phytosterol may also have a double bond in the structure. The number of double bonds is not particularly limited, and the sterol skeleton or the side chain thereof may have 1 or more, preferably 1 to 4, and more preferably 1 to 2 other double bonds.

Examples of the phytosterol include a 5-en-3-one body of phytosterol, a 4-en-3-one body of phytosterol, and a 4-en-3, 6-dione body of phytosterol.

Examples of the 5-en-3-one of phytosterols include campest-5-en-3-one, sitosteron-5-en-3-one, stigmast-5-en-3-one, brassicast-5-en-3-one, 24-alkylcholest-5, 7-dien-3-one, 24-alkylcholest-5, 8-dien-3-one, 24-alkylcholest-5, 9(11) -dien-3-one, 24-alkylcholest-5, 22-dien-3-one, 24-alkylcholest-5, 7, 22-trien-3-one, 24-alkylcholest-5, 8, 22-trien-3-one, 24-alkylcholest-5, 9(11), 22-trien-3-one and 24-alkylcholest-5, 25(27) -dien-3-one.

Examples of the 4-en-3-one body of phytosterol include campest-4-en-3-one, sitosterol-4-en-3-one, stigmast-4-en-3-one, and brassicast-4-en-3-one.

Examples of the 4-ene-3, 6-dione compounds of phytosterols include campest-4-ene-3, 6-dione, sitosterol-4-ene-3, 6-dione, stigmast-4-ene-3, 6-dione, brassicast-4-ene-3, 6-dione, 6-hydroxy-sitosterol-4-ene-3, 6-dione, 4-cholesten-3, 6-dione, campest-4-ene-3, 6-dione, and 24-alkylcholest-4-ene-3, 6-dione.

Examples of the "alkyl group" in each compound include a methyl group and an ethyl group.

As the phytosterol, commercially available phytosterol may be used, and phytosterol produced by a known chemical or biological method or a method based on the same may also be used. The biological method is a method using an enzyme, fermentation, or the like. Examples of commercially available phytosterols include stigmast-4-en-3-one manufactured by Sigma-Aldrich, and the like.

In some embodiments, as the phytosterol, a composition containing phytosterol is used. The composition containing phytosterol can be obtained, for example, by a method comprising fermenting a microorganism cultured with a medium containing phytosterol or a plant body containing phytosterol. Such methods are described in, for example, Japanese patent laid-open Nos. 2007-284348 and J.Nutr.Sci.Vitaminol5363-67.2006. In some embodiments, the phytosterol-containing composition comprises the 5-en-3-one bodies of phytosterols, the 4-en-3-one bodies of phytosterols, and the 4-en-3, 6-dione bodies of phytosterols. In some embodiments, the composition comprising phytosterols comprises 40% by weight or more (e.g., 40% by weight or more, 41% by weight or more, 42% by weight or more, 43% by weight or more, 44% by weight or more, 45% by weight or more, 46% by weight or more, or a combination thereof) of the 5-en-3-one body of phytosterols, the 4-en-3-one body of phytosterols, and the 4-en-3, 6-dione body of phytosterols,47 wt% or more, 48 wt% or more, 49 wt% or more, 50 wt% or more, 51 wt% or more, 52 wt% or more, 53 wt% or more, 54 wt% or more, 55 wt% or more, 56 wt% or more, 57 wt% or more, 58 wt% or more, 59 wt% or more, and 60 wt% or more). In some preferred forms, the phytosterol containing composition comprises: at least 1 5-en-3-one body of a phytosterol selected from the group consisting of campest-5-en-3-one, sitst-5-en-3-one, stigmast-5-en-3-one, brassicast-5-en-3-one, and combinations thereof; 4-en-3-one bodies of phytosterols selected from the group consisting of campest-4-en-3-one, sitst-4-en-3-one, stigmast-4-en-3-one, brassicast-4-en-3-one, and combinations thereof; and 4-ene-3, 6-dione bodies of phytosterols selected from the group consisting of campest-4-ene-3, 6-dione, sitosterol-4-ene-3, 6-dione, stigmast-4-ene-3, 6-dione, brassicast-4-ene-3, 6-dione, and combinations thereof. In other preferred embodiments, the phytosterol-containing composition comprises: sitosterol-5-en-3-one, sitosterol-4-en-3-one, campest-5-en-3-one, campest-4-en-3-one, and phytosterol-4-en-3, 6-dione (i.e., campest-4-ene-3, 6-dione, sitosterol-4-ene-3, 6-dione, stigmast-4-ene-3, 6-dione, and brassicast-4-ene-3, 6-dione).

Here, the components other than the phytosterol in the composition containing phytosterol are, for example, phytosterol (i.e., sitosterol, campesterol, stigmasterol, and brassicasterol) as a base.

Examples of phytosterols include campesterol, β -sitosterol, stigmasterol, brassicasterol, and 24-alkylcholesterol. The phytosterol may be used by obtaining it from a commercial supplier, or may be produced by a known method or a method based on the known method.

The "plant body containing phytosterol" may be the whole plant body containing phytosterol, or may be a part of the plant body (for example, stem, leaf, root, flower, fruit and seed). Examples of the plant containing phytosterol include all plants such as soybean, sunflower, rapeseed, sea buckthorn, pine, and the like.

Examples of the microorganism used for the culture include microorganisms exhibiting cholesterol oxidase activity. Examples of the microorganism include basidiomycetes (for example, Agaricus and Trametes versicolor), filamentous fungi (for example, Aspergillus and Monascus), and bacteria (for example, Arthrobacter, Streptomyces, Brevibacterium, Rhodococcus, and Bacillus). In some embodiments, specific examples of the microorganism include Arthrobacter Simplex (Arthrobacter Simplex, Nocardia alias (Nocardioides Simplex)), and the like.

As the microorganism, commercially available microorganisms can be used, or microorganisms having modified bacteriological properties obtained by artificial mutagenesis means (for example, EMS (ethyl methanesulfonate), diethyl sulfate, NTG (N-methyl-N' -nitro-N-nitrosoguanidine) and nitrosoguanidine), X-ray, gamma-ray and ultraviolet ray) can be used. Alternatively, as the microorganism, a transformed microorganism capable of expressing cholesterol oxidase by introducing a gene encoding cholesterol oxidase into a microorganism (e.g., escherichia coli) using a suitable vector and transforming the same can be used.

The amount of the phytosterol to be blended in the percutaneous absorbent can be suitably set, and is, for example, 0.1 to 20% by weight, preferably 0.5 to 10% by weight.

In the percutaneous absorbent, an alcohol (e.g., ethanol, isostearyl alcohol, cetyl alcohol, and phenoxyethanol), an emulsifier (e.g., polyglycerin fatty acid ester, lecithin derivative, polyethylene glycol fatty acid ester, sorbitan fatty acid ester, and polymer emulsifier), a percutaneous absorption enhancer (e.g., fatty acid, fatty alcohol, fatty acid ester, alkyl ether, aromatic organic acid, aromatic alcohol, aromatic organic acid ester, aryl ether, and ribose preparation), water (e.g., purified water, hot spring water, and deep water), an oil agent, a surfactant, a metal soap, a gelling agent, a powder, a water-soluble polymer, a film-forming agent, a resin, an ultraviolet ray protection agent, an encapsulating compound, an antibacterial agent, a perfume, an odor eliminating agent, a salt, a pH adjuster, a component used in medicines and cosmetics, and the like may be blended as necessary in addition to the phytosterol, Cooling agent, extract derived from animal/microorganism, plant extract, blood circulation promoter, astringent, antiseborrheic, whitening agent, antiinflammatory agent, active oxygen scavenger, cell activator, humectant, chelating agent, keratolytic agent, enzyme, hormone, vitamin, etc. These components may be used alone in 1 kind, or two or more kinds may be suitably combined and used, depending on the form of the percutaneous absorption preparation. For example, since phytosterol is insoluble in water, it can be dissolved in alcohols (e.g., ethanol) to make a liquid, or it can be made into a paste using an emulsifier. Some forms of the percutaneous absorption agent contain a percutaneous absorption enhancer.

Some forms of transdermal absorbents are drugs used to reduce subcutaneous fat. The drug may be applied to the skin, and may be in any of a liquid state or a semisolid state (e.g., a gel state, a paste state, and a paste state). Examples of the drug include gels, ointments, liquid preparations, patches, aerosols, aqueous ointments, and coatings.

Other forms of transdermal absorbents are cosmetics for reducing subcutaneous fat. The cosmetic may be applied to the skin, and may be in any of a liquid state or a semisolid state (e.g., a gel state, a paste state, and a paste state). As the cosmetics, for example, basic cosmetics (e.g., lotions, creams, emulsions, and lotions), hair cosmetics (shampoos, conditioners, and lotions), makeup cosmetics (e.g., foundation lotions and foundation lotions), and sunscreen cosmetics can be cited.

Some forms of transdermal absorbents for reducing subcutaneous fat are used in a manner suitable for the skin of a subject in need of subcutaneous fat reduction.

Further, herein, there is provided a method of reducing subcutaneous fat in a subject comprising applying phytosterols to the skin of a subject in need of such reduction. Alternatively, phytosterols for use in reducing subcutaneous fat are provided, which are used in a manner suitable for application to the skin.

The "subject" is a human, or an organism other than a human, such as a bird and a non-human mammal (e.g., cow, monkey, cat, mouse, rat, guinea pig, hamster, pig, dog, rabbit, sheep, and horse). The "subject" is preferably a human.

By "suitable for use" is meant that the phytosterol is applied to the skin of the subject in an amount sufficient to reduce the subcutaneous fat in the subject. In some embodiments, the phytosterol is applied to the skin on the subcutaneous tissue in a subject that has accumulated subcutaneous fat in need of reduction. It is preferable to apply the phytosterol to the skin of a part in which subcutaneous fat is likely to accumulate in a subject, for example, a part selected from the group consisting of upper arm, abdomen, and waist to thigh. The percutaneous absorption agent or the phytosterol can be applied to the skin by coating, sticking, spraying, etc., depending on the form of the agent.

The amount of the phytosterol used varies depending on age, sex, symptoms, frequency of administration, dosage form, etc. The amount of phytosterol is, for example, 5 mg/day to 1000 mg/day, or 50 mg/day to 500 mg/day, based on the number of administrations from 1 to a plurality of times per day.

In some preferred embodiments, the transdermal absorbent, method, or phytosterol may reduce subcutaneous fat prior to visceral fat.

In some embodiments, the transdermal absorbent, method, or oral administration of phytosterols may be used in combination with the oral administration of phytosterols.

In some embodiments, the transdermal absorbents, methods, or phytosterols may be used in combination with other pharmaceutically, or other cosmetically, active ingredients.

All documents and publications described in the present specification are incorporated in their entirety into the present specification by reference, regardless of the purpose thereof. The present specification includes the disclosure of the claims, the description and the drawings of japanese patent application No. 2018-161943 (application 8/30/8/2018), which is the basis of the priority claims of the present application.

Further, the objects, features, advantages and aspects of the present invention will be apparent to those skilled in the art from the description of the present specification, and the present invention can be easily implemented by those skilled in the art from the description of the present specification. The modes for carrying out the invention, specific examples, and the like are examples showing preferred embodiments of the present invention, and are shown for illustration or explanation, and the present invention is not limited to these. Various modifications can be made within the intent and scope of the invention disclosed in the specification based on the description of the present specification, and will be apparent to those skilled in the art.

Examples

Example 1: method for preparing phytosterol used as subcutaneous fat metabolism promoter

The phytosterol-containing composition as a test substance is produced by a method conforming to the fermentation method described in Japanese patent application laid-open No. 2007-284348. Specifically, a culture solution was prepared by culturing gram-positive bacteria in a medium containing phytosterol. Adding phytosterol into the prepared culture solution, layering the organic solvent, and then violently stirring by using a stirring blade. Then, the upper layer was recovered by double layer separation and dried to obtain a composition containing phytosterol. The obtained composition containing phytosterol contains more than 50 wt% of various sterones.

Sitosterol-5-en-3-one, sitosterol-4-en-3-one, campest-5-en-3-one, campest-4-en-3-one and phytosterol-4-en-3, 6-dione.

Here, the "phytosterol-4-ene-3, 6-dione" is a mixture of campest-4-ene-3, 6-dione, sitosterol-4-ene-3, 6-dione, stigmast-4-ene-3, 6-dione and brassicast-4-ene-3, 6-dione.

Analysis of various phytosterols and phytosterols was performed by high performance liquid chromatography according to the method described in Japanese patent application laid-open No. 2007-284348. Analytical values are as follows.

5-en-3-one bodies (12.5 wt%), 4-en-3-one bodies (36.6 wt%) and 4-en-3, 6-dione bodies (2.8 wt%). (total: 51.9 wt%)

The remaining components in the composition containing phytosterol are phytosterols (i.e., sitosterol, campesterol, stigmasterol, brassicasterol) as a base, and the like.

Campest-5-en-3-one as a test substance can be synthesized by using campesterol (TAMA BIOCHEMICAL CO., LTD.) as a raw material by the selective oxidation method of Parish et al or the Swern oxidation method (Parish E.J., et al: Synthetic communications (1992)22, p.2839-).

SD rats (Oriental Yeast co., Ltd.) were used which had been given a standard purified sample AIN-93G (Oriental Yeast co., Ltd.) for 2 weeks in advance for fattening. The abdominal hairs of the enlarged rats were removed by 20mm square, and the following substances were gently applied to each individual (n ═ 3 in each group) grouped as follows: 2ml of a composition containing phytosterol (30mg/ml ethanol solution) as a test substance, 2ml of campest-5-en-3-one (5mg/ml ethanol solution) as a test substance, or 2ml of an ethanol solution as a control. This procedure was continued for 2 weeks. After the coating test was completed, the rats were dissected, and liver fat, peritestis fat, perirenal fat, mesenteric fat and subcutaneous fat were carefully recovered and the weights thereof were measured.

As a result, the amount of subcutaneous fat was significantly reduced (P <0.05) and reduced by about 40% when the composition (P) containing phytosterol was applied, compared to the case when the control (E) was applied (fig. 1). In addition, for the amount of subcutaneous fat, the subcutaneous fat was also significantly reduced (p <0.05) in the case of coating campest-5-en-3-one (C) and the amount of subcutaneous fat was reduced by about 16% compared to the case of coating control (E).

Thus, the subcutaneous fat when coated with the composition containing phytosterols or campest-5-en-3-one was reduced by 40% and 16%, respectively, compared to the control. The amount of phytosterol administered on a transdermal basis when using a composition containing phytosterol is about 31mg per day (30mg/ml x 2ml x 0.519) for a total of about 434mg for 2 weeks. In addition, the dose based on transdermal administration when campest-5-en-3-one is used is about 10mg per day (5 mg/ml. times.2 ml), and is only about 140mg in total for 2 weeks.

On the other hand, it is reported that the subcutaneous fat reduction rate of campest-5-en-3-one, which significantly reduces visceral fat when orally administered at about 120 mg/day within 8 weeks, is 26% (obesity study (2007),13(3), p.244-249). Based on the amount of campest-5-en-3-one administered orally, about 120mg per day, thus about 6720mg in total for 8 weeks.

From these results, it can be seen that: when phytosterol is administered transdermally, subcutaneous fat is reduced in a short period of time and in a small amount as compared with the case of oral administration.

Here, it is generally known that visceral fat is easily reduced, and subcutaneous fat is not easily reduced. It is also known that subcutaneous fat metabolism is often performed after visceral fat is metabolized, and that subcutaneous fat metabolism is preferentially performed during aerobic exercise. Fat in the perimesentery, perikidney, peritestis, liver and the like is called visceral fat. However, the amount of fat at this site was not significantly reduced when the test substance was applied as compared to the case of the control (fig. 2 to 5). Namely, it can be seen that: when the test substance is applied, subcutaneous fat is reduced earlier than visceral fat.

Here, the significant difference test in the present example was performed by first calculating the mean ± standard error of each group and performing the variance test. In addition, significant difference tests were performed using t-test for the case of homoscedasticity and Welch method for the case of heteroscedasticity.

According to the present embodiment, it was confirmed that: when phytosterol is administered transdermally, subcutaneous fat is reduced in a short period of time and in a small amount as compared with the case of oral administration.