JP2016188180A - Composition containing specific ingredients - Google Patents

Abstract

An object of the present invention is to provide a composition exhibiting a UCP1 expression promoting action and / or a brown adipocyte differentiation promoting action, which is free from problems of side effects and can be ingested for a long time. . The object is to contain at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint, and to promote UCP1 expression and / or brown adipocytes. It is solved by a composition having a differentiation promoting action. [Selection figure] None

Description

The present invention relates to a composition containing a specific component and having a UCP1 expression promoting action and / or a brown adipocyte differentiation promoting action.

Uncoupling proteins (UCPs) have a function of uncoupling oxidative phosphorylation in the inner mitochondrial membrane and dissipating energy as heat (see Non-Patent Document 1). In particular, UCP1 is specifically expressed in brown adipocytes, which are heat production sites, and plays a role in converting neutral lipids to heat and directly consuming energy without undergoing ATP synthesis in the electron transport system. Bear. Moreover, if differentiation from progenitor cells to brown adipocytes can be promoted, the expression level of UCP1 can be increased, and neutral lipids can be converted into heat. Therefore, if the UCP1 expression level or brown adipocyte differentiation can be promoted, there is a possibility that obesity can be resolved through consumption of neutral lipids. Patent Document 1 describes such a brown adipocyte differentiation promoting agent and UCP1 expression promoting agent having catechins as an active ingredient.

JP 2014-65672 A

124th Japan Medical Society Symposium Record, p. 62-70

A substance capable of promoting the expression level and activation of UCP1 described in Patent Document 1 contains a specific substance isolated and purified from a natural product as an active ingredient. However, since such a specific substance has a strong pharmacological action, it is necessary to be careful when taking it, and there is a risk of causing side effects due to excessive intake.

The present invention has been made in view of such circumstances, and has a UCP1 expression promoting action and / or brown adipocyte differentiation promoting action, which has no problem of side effects and can be ingested for a long time and continuously. Providing a product is a problem to be solved by the invention.

The present inventors have intensively studied to solve the above problems, and surprisingly, by containing a specific component, a particularly remarkable UCP1 expression promoting effect and / or brown adipocyte differentiation promoting effect is obtained. I found out that And as a composition for promoting UCP1 expression, a composition for promoting metabolism, an anti-obesity composition, a composition for reducing body fat, and a composition for promoting heat production, the inventors succeeded in creating a composition containing specific components. The present invention has been completed on the basis of such knowledge and successful examples.

Therefore, according to the present invention, there is provided a cosmetic composition containing at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

According to another aspect of the present invention, a UCP1 expression promoting composition containing at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint is provided.

According to another aspect of the present invention, there is provided a composition for activating brown adipocytes containing at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint. The

According to another aspect of the present invention, a composition for promoting metabolism comprising at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint is provided.

According to another aspect of the present invention, there is provided an anti-obesity composition containing at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

According to another aspect of the present invention, there is provided a composition for reducing body fat containing at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

According to another aspect of the present invention, there is provided a composition for promoting heat production containing at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

According to another aspect of the present invention, there is provided a composition containing at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.

The composition of the present invention has a UCP1 expression promoting action and / or brown adipocyte differentiation promoting action, and in addition to these actions, brown adipocyte activation action, metabolism promoting action, anti-obesity action, body fat It can be expected to show a reducing action, a heat production promoting action, a UCP1 expression promoting action, an energy production promoting action, a neutral lipid lowering action, a sebum suppressing action, a fat metabolism promoting action and the like.

Since the specific component used in the composition of the present invention has a track record of use in pharmaceuticals and cosmetics, the composition of the present invention is highly safe. Therefore, the composition of the present invention is useful as a composition for promoting UCP1 expression, a composition for promoting metabolism, a composition for anti-obesity, a composition for reducing body fat, and a composition for promoting heat production by having the above-described action. And can be expected to be provided in parenteral or oral form.

Hereinafter, the present invention will be described in detail.
The composition of the present invention has a UCP1 expression promoting action and / or a brown adipocyte differentiation promoting action, and is a component of any one of hop, trehalose, aspartame, barley, ceramide, placenta, alanine and mint (hereinafter, specified) At least).

The composition of the present invention may have any one of UCP1 expression promoting action and brown adipocyte differentiation promoting action, or may have all these two kinds of actions. Good. Further, the specific component may be any one of the specific components or two or more components.

In addition to the UCP1 expression promoting action and brown adipocyte differentiation promoting action, the composition of the present invention has brown adipocyte activation action, metabolism promoting action, anti-obesity action, body fat reducing action, heat production promoting action, energy production promoting. An action, a neutral lipid lowering action, a sebum suppressing action, and a fat metabolism promoting action may be exhibited.

Since the composition of the present invention has the above-described action, the composition of UCP1 expression promoting composition, composition for promoting metabolism, composition for anti-obesity, composition for reducing body fat, and composition for promoting heat production It can take.

The hop is not particularly limited as long as it is known as Humulus lupulus , and for example, it is possible to use parts such as flowers (carp flowers, cones), seeds, fruits, leaves, stems and roots. However, it is preferable to use hop cones.

Mint is not particularly limited as long as it is known as Mentha piperita , and for example, its parts such as flowers, seeds, fruits, leaves, stems, roots can be used. It is preferable to use it.

Barley is not particularly limited as long as it is known as barley ( Hordeum vulgare ), and for example, the parts such as flowers, seeds, fruits, leaves, stems and roots can be used. Is preferably used.

The placenta is not particularly limited as long as it is a known placenta, for example, an extract derived from placenta such as pig, human, cow, horse, sheep, or animal placenta which is a component thereof, fish ovarian membrane, etc. Extracts derived from the above, or marine placenta that is a component thereof, extracts derived from plant germs, or plant placenta that is a component thereof, and animal placenta are preferred. Is more preferable.

The ceramide is not particularly limited as long as it is a compound in which sphingosine and a fatty acid are amide-bonded.For example, ceramide derived from a natural product or ceramide obtained by synthesis is preferable, but a ceramide derived from a natural product is preferable, rice, Ceramide derived from corn, konjac, maitake, tamagotake, soybean, wheat, beet and the like is more preferred, and ceramide derived from rice (glucosylceramide) is more preferred.

The trehalose is not particularly limited as long as it is a known trehalose. For example, trehalose itself and trehalose derivatives such as hydrates and optical isomers thereof can be mentioned, and preferably D-(+)-trehalose. is there.

The aspartame is not particularly limited as long as it is a known aspartame, and examples thereof include aspartame derivatives such as aspartame itself and optical isomers thereof, preferably aspartame whose CAS registration number is 22839-47-0. It is.

The alanine is not particularly limited as long as it is a known alanine, and examples thereof include derivatives such as alanine itself and optical isomers thereof, preferably alanine having a CAS registration number of 338-69-2. is there.

Among the specific components, natural products may be processed products. Examples of the processed product include, but are not limited to, a dry powder, a fragmented product and a dry powder thereof, a juice and a dry powder thereof, an extract and a dry powder thereof. However, from the viewpoints of ease of processing, storage, transportation, etc. and versatility of usage forms, a dry powder is preferable.

A method for obtaining a dried natural product is not particularly limited. For example, barley foliage powder can be produced by a method disclosed in Japanese Patent No. 3277181. That is, barley stalks and leaves are harvested, washed with water, washed away with mud, drained, and then cut into appropriate lengths. Next, blanching treatment such as hot water treatment or steam heat treatment is performed as necessary. What is necessary is just to determine suitably the temperature and time of the process at this time according to the quantity of the barley foliage to process, and the pH of hot water. The blanched barley foliage is immediately cooled and dehydrated by centrifugation or the like. Next, drying is performed so that the water content is 5 wt% or less. Furthermore, barley foliage powder can be obtained through a coarse pulverization step, a heating step, a fine pulverization step, and the like.

The specific component is a natural product that has been ingested by humans over a long period of time, its derivatives and synthetic products, and has high safety. Therefore, the composition of the present invention has high practicality, as it is, or By processing, it can be applied to various uses in a parenteral or oral form.

The specific component may be produced by a method generally known by those skilled in the art or may be commercially available.

The specific component exhibits a UCP1 expression promoting action or a brown adipocyte differentiation promoting action. In the present specification, “expression promoting action” means, for example, in the case of UCP1 expression promoting action, promoting the expression level of UCP1 gene in a subject, increasing the translation amount of UCP1 protein, and activating UCP1 Of at least one of them. The brown adipocyte activation action means at least one action of activating brown adipocytes and increasing the number of brown adipocytes.

Regarding the UCP1 expression promoting action, as described in Non-Patent Document 1, the function of UCP1 is decreased in obese animals, the amount of UCP1 is increased in animals that are not obese even after eating, artificially UCP1 It is known that mice with a reduced expression of the expression are obese and high-expressing mice are thin. Therefore, an anti-obesity effect can be expected by the UCP1 expression promoting action, and substances that actually have UPC1 expression promoting action promote lipolysis in white adipocytes and at the same time activate UCP1 to convert the released fatty acid into heat. Can ultimately reduce body fat.

The heat production promoting action refers to, for example, the action of promoting heat production from neutral lipids by brown adipocytes. The neutral lipid lowering action refers to, for example, an action of degrading neutral lipids to reduce the total amount of neutral lipids in vivo. The sebum suppressing action refers to, for example, an action of suppressing the generation of sebum. The fat metabolism promoting action means, for example, an action of qualitatively converting fat cells and adipose tissue forming an obese state into small normal fat cells. The body fat reducing action refers to, for example, an action of reducing body fat by changing a released fatty acid into heat. The metabolism promoting action and the anti-obesity action refer to, for example, an action of promoting metabolism so as to reduce body fat and an action of suppressing obesity by the action. The energy production promoting action refers to an action of producing energy through heat production from neutral lipids by brown adipocytes, for example. Each action in this paragraph includes an action having the meaning described above, but also includes those realized by other action mechanisms.

The anti-disease effect exhibited by the anti-disease composition such as the anti-obesity composition of the present invention is, for example, the present or future obesity or obesity in the subject in the case of the anti-obesity effect exhibited by the anti-obesity composition. It means to suppress or delay the improvement of the state. Insulin resistance refers to a state in which the sugar absorption promoting action, which is the main action of insulin, is weakened in the liver, fat cells, skeletal muscles, and the like. The anti-insulin resistance effect is to suppress or delay or improve the value of a current or future index of insulin resistance by SSPG (Steady-state plasma glucose) method in a subject. Say.

Obesity is known to cause various diseases. Examples of such diseases include type 2 diabetes, hypertension, and hyperlipidemia. In addition, these diseases can lead to stroke, ischemic heart disease, and the like. In an obese state, fat cell hypertrophy is observed, TNF-α and free fatty acid (FFA) are secreted, which inhibits glucose uptake in insulin target cells such as muscle cells and liver cells, and insulin Secretion of adiponectin, which promotes the action of, is suppressed, causing a decrease in insulin sensitivity at the cellular level, resulting in insulin resistance. The diseases caused by obesity are viewed as a series of metabolic abnormalities based on insulin resistance. Therefore, by using the composition of the present invention, it is possible to prevent or improve diseases such as type 2 diabetes, hypertension and hyperlipidemia associated with obesity and obesity.

Here, prevention and improvement of type 2 diabetes refers to suppression or delay of becoming a diabetic state or borderline state, or approaching a state called a normal range from those states. Prevention and improvement of hypertension refers to suppression or delay of becoming a state of high blood pressure or a boundary region, or approaching a state called a normal region from that state. Prevention and improvement of hyperlipidemia refers to suppression or delay of becoming a hyperlipidemia state or boundary state, or approaching a state called a normal region from that state.

The manufacturing method of the composition of this invention is not specifically limited, For example, it can be set as a solid composition by containing a specific component under room temperature or a heating. Alternatively, this solid composition can be dissolved in a solvent such as water to form a liquid composition. Furthermore, it can be set as a liquid composition by adding and mixing another solid component with the liquid of a specific component. In addition, you may pulverize the obtained liquid composition through a drying process. Examples of the drying process in this case include spray drying and freeze drying, but are not limited thereto.

The composition of the present invention can be used as it is or mixed with other components depending on the application. Thus, the composition of this invention can mix | blend various things other than a specific component, as long as the objective of this invention can be achieved.

For example, the composition of the present invention can further contain additives used in ordinary processing such as excipients, extenders, binders, thickeners, emulsifiers, colorants, fragrances, perfume oils. . The amount of the additive used is not particularly limited as long as it does not hinder the solution of the problems of the present invention, and is appropriately adjusted.

The composition of the present invention is not particularly limited as long as it is a commonly used form, for example, liquid, lotion, mousse, gel, emulsion, suspension, cream, ointment, sheet, aerosol , Sprays, sticks, powders, granules, granules, tablets, rods, plates, blocks, solids, pastes, capsules, caplets, and the like.

The composition of the present invention can be used as a parenteral composition, for example, in a form suitable for cosmetics. That is, one aspect of the composition of the present invention is a cosmetic composition. For example, the composition of the present invention can be processed into various forms such as lotions, emulsions, gels, creams, ointments, etc. as they are or mixed with additives usually used in cosmetic processing. . Specifically, it can be used as a lotion, cosmetic cream, milky lotion, cream, pack, hair tonic, hair cream, shampoo, hair rinse, treatment, facial cleanser, foundation, hair restorer, aqueous ointment, spray and the like.

The content of the specific component contained in the composition of the present invention is not particularly limited as long as UCP1 expression promoting action and / or brown adipocyte differentiation promoting action is recognized, but as an oral composition, for example, It is 0.0001 wt% or more, preferably 0.001 wt% or more with respect to the whole composition, and as a parenteral composition such as cosmetics, for example, 0.00001 wt% or more, preferably 0.0001 wt% or more. is there.

As a specific aspect of the composition of the present invention, for example, a composition containing 0.001 to 100 wt% of any one of the specific components with respect to the entire composition can be mentioned. Among these, a composition containing a specific component at 0.01 to 80 wt% is more preferable.

Another specific embodiment of the composition of the present invention includes a composition containing any two of the specific components in a proportion of 10 to 100 wt% in total with respect to the entire composition. Among these, the composition which contains 10-90 wt% of one component and 90-10 wt% of the other component among these 2 types of components is mentioned.

Another specific embodiment of the composition of the present invention includes a composition containing any three of the specific components in a total proportion of 10 to 100 wt% with respect to the entire composition. Among these, the composition which contains 10 to 50 wt% of the first component among the three components, 30 to 40 wt% of the second component, and 60 to 10 wt% of the third component.

Hereinafter, formulation examples according to specific embodiments of the composition of the present invention will be shown. However, the present invention is not limited to these formulation examples, and the present invention has various modes as long as the problems of the present invention can be solved. Can take.

(Formulation example 1: lotion)
100 parts by mass as a whole, 0.01 parts by mass of trehalose, 0.01 parts by mass of ginger extract, 0.01 parts by mass of ketsumeishi, 10 parts by mass of glycerin, 3 parts by mass of diglycerin, 12 parts by mass of 1,3-butylene glycol , 3 parts by weight of pentylene glycol, 0.1 part by weight of sodium hyaluronate, 0.01 part by weight of citric acid, 0.02 part by weight of sodium citrate, 0.1 part by weight of xanthan gum, 0.15 part by weight of methyl paraben, carbomer 0 The composition of the present invention was prepared in the form of a lotion by mixing 2 parts by mass, 0.03 part by mass of sodium hydroxide and the remaining part of water.

(Formulation example 2: shampoo)
The total amount is 100 parts by mass, hop extract 0.02 parts by mass, caffeine 0.01 parts by weight, sodium laureth sulfate 7.5 parts by mass, cocamidopropyl betaine 4.2 parts by mass, cocamide DEA 3 parts by mass, , 3-butylene glycol 0.1 parts by weight, polyquaternium-10 0.225 parts by weight, citric acid 0.15 parts by weight, sodium citrate 0.05 parts by weight, phenoxyethanol 0.9 parts by weight and water balance The composition of the present invention was prepared in the form of a shampoo.

(Formulation example 3: soap)
100 parts by weight as a whole, 0.5 parts by weight of aspartame, 0.2 parts by weight of Merirot extract, 0.1 parts by weight of elastin, 2 parts by weight of glycerin, 1 part by weight of olive oil, 0.1 parts by weight of EDTA-4 sodium, The composition of the present invention was prepared in the form of a soap by mixing and solidifying etidronate tetrasodium 0.2 parts by mass and soap residue.

(Formulation example 4: emulsion)
100 parts by mass as a whole, 0.1 part by mass of ceramide, 0.1 part by mass of alanine, 0.1 part by mass of arginine, 3 parts by mass of sucrose fatty acid ester, 12 parts by mass of glycerin, 6 parts by mass of squalane, 24 parts of dimethyl silicone oil Emulsion mode by mixing parts by mass, 1 part by mass of polypropylene glycol, 0.06 parts by mass of thickener, 0.2 parts by mass of phenoxyethanol, 5 parts by mass of ethanol, 0.01 parts by mass of sodium hydroxide and the remainder of purified water The composition of the present invention was prepared.

(Formulation Example 5: Cosmetic cream)
100 parts by weight as a whole, 0.1 parts by weight of alanine, 0.1 parts by weight of fructooligosaccharide, 15.0 parts by weight of squalane, 4.0 parts by weight of octyldodecyl myristate, 0.2 parts by weight of hydrogenated soybean phospholipid, Butyl alcohol 2.4 parts by mass, hardened oil 1.5 parts by mass, stearic acid 1.5 parts by mass, lipophilic glyceryl monostearate 1.5 parts by mass, polyglyceryl monostearate 0.5 parts by mass, behenyl alcohol 0. 8 parts by weight, polyglyceryl monomyristate 0.7 part by weight, beeswax 0.3 part by weight, d-δ-tocopherol 0.1 part by weight, methylparaben 0.3 part by weight, C10-30 alkyl-modified carboxyvinyl polymer 0. 2 parts by mass, carboxyvinyl polymer 0.1 part by mass, 1,3-butanediol 18.0 parts by mass, water By mixing sodium 0.1 parts by mass of purified water balance of the compositions of the present invention was prepared in the manner of cosmetic creams.

(Formulation Example 6: Packing agent)
100 parts by weight as a whole, placenta 0.1 parts by weight, gallic acid 0.01 parts by weight, polyvinyl alcohol 20.0 parts by weight, glycerin 5.0 parts by weight, ethanol 20.0 parts by weight, kaolin 6.0 parts by weight The composition of the present invention was prepared in the form of a pack agent by mixing 0.2 parts by mass of a preservative, 0.1 part by mass of a fragrance, and the remainder of purified water.

(Formulation Example 7: Tablet)
100 parts by weight as a whole, 10 parts by weight of barley young leaf powder, 8 parts by weight of ginger extract, 5 parts by weight of caffeine, 5 parts by weight of mellote, 20 parts by weight of crystalline cellulose, 50 parts by weight of lactose, 4 parts by weight of magnesium stearate The composition of the present invention was prepared in the form of a tablet by mixing and tableting the remainder of the corn starch.

(Formulation Example 8: Tablet)
100 parts by weight as a whole, 0.5 parts by weight of alanine, 8 parts by weight of ketsumeishi powder, 5 parts by weight of merirot, 0.5 parts by weight of fructooligosaccharide, 20 parts by weight of crystalline cellulose, 50 parts by weight of lactose, 4 parts by weight of magnesium stearate Part and corn starch The composition of the present invention was prepared in the form of a tablet by mixing and tableting the remainder.

(Formulation example 9: granule)
By mixing and granulating 100 parts by weight of the whole, 15 parts by weight of hop powder, 5 parts by weight of Ketsumeishi extract, 10 parts by weight of lactose, 1 part by weight of calcium stearate and the remaining part of crystalline cellulose, this is used in the form of granules. Inventive compositions were prepared.

(Formulation example 10: capsule)
The total amount is 100 parts by mass, and 10 parts by mass of mint extract, 20 parts by mass of ginger extract, 5 parts by mass of elastin, 8 parts by mass of lecithin and the remainder of olive oil are used as the content liquid. By encapsulating, the composition of the present invention was prepared in the form of a capsule.

(Formulation example 11: liquid agent)
Based on 100 parts by mass of the whole, 0.84 parts by mass of barley young leaf extract, 1 part by mass of arginine, 1 part by mass of alanine, 10 parts by mass of fructose glucose liquid sugar, 1 part by mass of citric acid, 0.02 part by mass of sodium benzoate, A composition of the present invention was prepared in the form of a liquid by mixing 2 parts by mass of a fragrance preparation, 0.05 part by mass of sucralose, 0.03 part by mass of acesulfame potassium, and the remainder of purified water.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to these examples, and the present invention can take various modes as long as the problems of the present invention can be solved. .

It was demonstrated as follows that the specific components trehalose, aspartame, barley, ceramide, placenta, alanine, mint and hop have a remarkable remarkable UCP1 expression promoting action.

(1) Among the test substance specific components, commercially available trehalose, aspartame, ceramide, placenta, alanine, mint and hop were used.

Barley used the dry powder of barley foliage prepared as follows. That is, the barley stalks and leaves harvested at a height of about 30 cm were washed with water to remove the adhering mud and the like, and a pretreatment was performed to cut the size to about 5 to 10 cm. The pretreated stalks and leaves were blanched only once with hot water at 90 to 100 ° C. for 90 to 120 seconds, and then cooled with cold water. Subsequently, the obtained foliage was dried with warm air at 80 ° C. to 130 ° C. for 20 minutes to 180 minutes in a dryer until the water content became 5% by mass or less. The dried foliage was coarsely pulverized to a size of about 1 mm using a mixer. The obtained barley stalks and leaves were finely pulverized using a pulverizer so that 90% or more passed through the 200 mesh section to obtain dry powder of barley stalks and leaves.

(2) Preparation of collagenase solution A collagenase solution was prepared by shaking a DMEM medium prepared so that collagenase was 0.2% (W / V) and BSA was 1% (W / V) at room temperature for 1 hour. Prepared.

(3) Preparation of Brown Adipocyte Solution After euthanizing ICR mice (Kyudou) with diethyl ether, brown adipose tissue was removed from the back. From the obtained brown adipose tissue, adipose tissue, blood vessels, muscles and the like adhering to the periphery of the brown fat were removed using tweezers, and then the brown fat was washed 3 times in DMEM. Subsequently, the brown fat was immersed in the collagenase solution, and the brown fat was subjected to a shredding process in which the brown fat was finely chopped with scissors. The cell fat dispersion was obtained by incubating the shredded brown fat in a collagenase solution at 37 ° C. for 1 hour. The obtained cell dispersion was filtered with a cell strainer having a pore size of 100 μm, and then subjected to centrifugation (800 rpm, 5 min, 20 ° C.). The precipitated cells were then suspended in DMEM to obtain a cell suspension. Subsequently, the obtained cell suspension was filtered using a 25 μm filter sterilized by autoclaving. The filtered supernatant was centrifuged (800 rpm, 5 min, 20 ° C.), and the precipitated cells were suspended in DMEM containing 10 vol% FBS to obtain a brown adipocyte liquid.

(4) Preparation of medium DMEM containing 10 vol% FBS was used as a growth medium.

As a differentiation induction medium, a growth medium containing 2.5 μM dexamethasone, 10 μg / ml insulin and 0.5 mM IBMX was prepared.

As the maintenance medium, the above insulin solution was used to prepare a growth medium containing 10 μg / ml insulin.

The test substance-containing medium was prepared as follows. That is, each test substance was dissolved in DMSO, diluted with a maintenance medium, prepared so that the final concentration of DMSO was 0.5 vol%, and filter sterilized. After sterilization, a predetermined concentration, that is, hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint was diluted to 20 μg / ml using a maintenance medium containing 0.5 vol% DMSO.

(5) Place 5 ml of collagen coat solution in a 75 cm ² flask of cell culture, leave it at room temperature for about 1 hour, and then remove the collagen coat solution by suction and wash twice with 5 ml of PBS to obtain a collagen coat flask. It was.

The brown adipocyte solution was added to the collagen-coated flask, and the brown adipocytes were cultured for 24 hours in a 37 ° C., 5% CO ₂ incubator. Next, the cultured brown adipocytes were washed once with a growth medium, then replaced with a new growth medium, and cultured until subconfluent.

The cultured brown adipocytes were subjected to trypsin treatment and suspended. The obtained floating cells were seeded in each well of a collagen-coated (collagen coating solution 350 μL / well, PBS wash 350 μL / well × 2 times).

Brown adipocytes were cultured using a growth medium until confluent. After removing the medium from each well, 500 μl of the differentiation induction medium was added to each well and cultured for 48 hours.

After removing the differentiation induction medium, 500 μl of a 20 μg / ml test substance-containing medium was added and cultured for 6 days. The medium was changed every other day. As a control, 500 μl of a medium not containing a test substance was added and cultured for 6 days. The medium was changed every other day.

After culturing for 8 days, the supernatant was collected, RNA was collected using RNeasy Mini Kit (Qiagen), and cDNA was synthesized using QuantTect Reverse Transfer Kit (Qiagen).

Quantitative real-time PCR was performed with Rotor-Gene SYBR Green PCR Kit (Qiagen) using the obtained cDNA as a template and a primer for UCP1 gene (Qiagen), and the mRNA expression level of UCP1 was measured. As an endogenous control, GAPDH mRNA expression level was measured using a GAPDH primer (Qiagen).

(6) Evaluation results of mRNA expression level of UCP1 when each test substance of hop, trehalose, aspartame, barley, ceramide, placenta, alanine and mint was provided (control mRNA expression level is 1.00) Table 1 shows the relative expression levels at the time.

As Table 1 shows, when hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint were used as test substances, the expression level of UCP1 was large.

The composition of the present invention contains a specific component applicable to both parenteral and oral forms, and is particularly useful for a subject suffering from cellulite, obesity or a disease associated therewith. Contributes to the health and welfare of such subjects.

Claims (8)

A cosmetic composition comprising at least one ingredient selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint. A composition for promoting UCP1 expression comprising at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint. A composition for activating brown adipocytes, comprising at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint. A composition for promoting metabolism comprising at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint. An anti-obesity composition containing at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint. A composition for reducing body fat, comprising at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint. A composition for promoting heat production comprising at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint. A composition comprising at least one component selected from the group consisting of hops, trehalose, aspartame, barley, ceramide, placenta, alanine and mint.