JP7445689B2 - lipolysis accelerator - Google Patents

Description

The present invention relates to a lipolysis promoter.

Body fat is produced when the intake of energy exceeds the energy consumed, and the excess is accumulated as neutral fat in white adipocytes. Obesity results from accumulation of excess fat, and as a result, obesity not only causes various lifestyle-related diseases but also poses a major problem in terms of beauty. Obesity, in which a large amount of body fat is stored as visceral fat, has been linked to pathological conditions such as insulin resistance and arteriosclerosis, and obesity, in which a large amount of body fat is stored as subcutaneous fat, is of great interest to both men and women from a beauty perspective. It's happening.

Conventionally, various measures have been taken to prevent and improve obesity, such as restricting energy intake, such as dietary restrictions and searching for substances that inhibit carbohydrate absorption in the gastrointestinal tract. However, restricting energy intake also causes a decrease in basal metabolic rate, and obesity may not be improved. In addition, limiting energy intake can lead to a lack of necessary nutrients, which can harm health. Therefore, in order to eliminate obesity, it has come to be considered that it is ideal to actively decompose accumulated fat and dissipate it as heat energy. Under these circumstances, in recent years, there has been an active search for functional ingredients that have a lipolysis-promoting effect from food materials, and many lipolysis promoters and food and drink products have been proposed.

For example, a mixture prepared by combining salmon milt extract, brewer's yeast extract, barley grass extract, and chicken collagen (chicken collagen) has an excellent lipolysis promoting effect (Patent Document 1: JP-A-2011-074051 ), an extract of birch of the family Betulaceae, and an extract of Kumaza of the family Poaceae improve obesity and prevent the increase of adipose tissue by promoting the decomposition of fat in whole body or local adipose tissue (patented patent) Document 2: JP 2006-045120), specific plants such as Japanese juniper or their extracts promote the decomposition of neutral fats accumulated in adipose tissue and exhibit slimming effects such as suppressing, preventing, or improving obesity. It is known that it is useful as a medicine, food, or cosmetic (Patent Document 3: Japanese Patent Laid-Open No. 2012-229266). However, at present, the lipolytic effects of extracts of these plants are not sufficient. Furthermore, Patent Document 4 discloses that a composition in which a licorice hydrophobic extract and an antioxidant component are combined has the effects of suppressing body fat accumulation, promoting body fat decomposition, and promoting energy production. Hydrophobic extracts alone do not have a sufficient lipolysis promoting effect.

JP2011-074051 JP2006-045120 JP2012-229266 International Publication No. 2008/143182

The problem to be solved by the present invention is to provide a novel lipolysis promoter that has an excellent lipolysis promoting effect.

As a result of extensive research, the present inventors discovered that "gliasperin B", a specific component in the hydrous ethanolic extract of licorice, has an excellent lipolysis promoting effect, and in order to complete the present invention. It's arrived. The gist of the present invention is as follows.

[1] A lipolysis promoter containing gliasperin B as an active ingredient.
[2] The lipolysis promoter according to [1], wherein the gliasperin B is derived from a licorice extract.
[3] The lipolysis promoter according to [2], wherein the licorice extract is a hydrous ethanol extract of licorice.
[4] The lipolysis promoter according to [3], wherein the ethanol concentration in the water-containing ethanol is 0.1% (v/v) to 99.9% (v/v).
[5] The lipolysis promoter according to [4], wherein the ethanol concentration in the water-containing ethanol is 30% (v/v) to 70% (v/v).
[6] The lipolysis promoter according to any one of [1] to [5], wherein the content of gliasperin B is 0.00001% by mass to 50% by mass.

The lipolysis promoter of the present invention exhibits an excellent lipolysis promoting effect by containing gliasperin B as an active ingredient. By using the lipolysis promoter of the present invention, fat in fat cells can be efficiently decomposed and body fat such as subcutaneous fat and visceral fat can be reduced. Therefore, according to the lipolysis promoter of the present invention, obesity and lifestyle-related diseases can be prevented and improved over the long term, and health can be maintained and improved.

FIG. 2 is a diagram showing the lipolysis promoting effects of licorice extract and gliasperin B. It is a figure showing the effect of licorice extract on body weight, BMI, and visceral fat level.

Below, the lipolysis promoter of the present invention will be explained in detail.

<Lipolysis accelerator>
The lipolysis promoter of the present invention contains gliasperin B as an active ingredient. In addition to the essential component gliasperin B, the lipolysis promoter of the present invention may contain other components as long as the effects of the present invention are not impaired. Below, gliasperin B and other components contained in the lipolysis promoter of the present invention, the form of the lipolysis promoter of the present invention, etc. will be explained.

(Gliasperine B)
Glyasperin B (hereinafter also referred to as "GB") is an isoflavanone derivative represented by the following formula. 3-(2,4-dihydroxyphenyl)-5-hydroxy-2,3-dihydro-6-(3-methyl-2-butenyl)-7-methoxy-4H-1-benzopyran-4-one, 3-( 2,4-dihydroxyphenyl)-5-hydroxy-6-(3-methyl-2-butenyl)-7-methoxy-3,4-dihydro-2H-1-benzopyran-4-one, or 3-(2, It is also represented as 4-dihydroxyphenyl)-5-hydroxy-7-methoxy-6-(3-methylbut-2-en-1-yl)-3,4-dihydro-2H-1-benzopyran-4-one.

Gliasperin B is a component that has been confirmed to be contained in licorice extract, and gliasperin B in the present invention is preferably derived from licorice extract. Until now, it has been known that licorice extract has a lipolysis-promoting effect, and there are documents suggesting that glabridin, which is abundant in licorice extract, may have a lipolysis-promoting effect (Patent Document 4, etc.) reference). However, it was not clear which component in licorice extract exerts the effect of promoting lipolysis. This time, the present inventors discovered for the first time that among the many components present in licorice extract, gliasperin B has a lipolysis promoting effect, and invented a lipolysis promoter containing this gliasperin B as an active ingredient. Completed.

Gliasperin B in the present invention may be derived from licorice or may be a chemically synthesized product.

The type of licorice is not particularly limited, but includes plants of the genus Glycyrrhiza of the Fabaceae family, such as Glycyrrhiza uralensis (G. uralensis Fisch. et DC; Glycyrrhiza inflata), Glycyrrhiza inflata (BAT.), G. glabra L., G. glabra L. var grandu rifera Regel and Herder, G. aspera, G. echinata L.; Chinese daylily) , G. pallidiflora Maxim (G. pallidiflora Maxim), and the like.

A method for obtaining gliasperin B derived from licorice includes a method of purifying it from a licorice extract. Specifically, the licorice extract is a licorice extract obtained by extracting the whole licorice plant or a part (e.g. roots, stems, stolons, leaves, flowers, fruits, etc.) of pulverized products with a solvent. The licorice extract may be spray-dried or freeze-dried. Examples of the extraction solvent include water, alcohols such as methanol and ethanol, and mixed solvents of water and alcohols or ketones such as acetone. Among these, the extraction solvent is preferably water, alcohol, or hydrous alcohol, and more preferably hot water, ethanol, or hydrous ethanol.

The alcohol concentration of the hydrous alcohol is 0.1% to 99.9% by mass, preferably 10% to 99.9% by mass, more preferably 30% to 70% by mass. , more preferably 40% by mass to 60% by mass, particularly preferably 50% by mass. Alternatively, it is 0.1% (v/v) to 99.9% (v/v), preferably 10% (v/v) to 99.9% (v/v), and 30% ( It is more preferably from 40% (v/v) to 60% (v/v), and even more preferably from 50% (v/v) to 70% (v/v). It is particularly preferable.

Distilled water is added to and dissolved in the dried licorice extract to prepare an aqueous solution of licorice extract. Add half the amount of organic solvent such as ethyl acetate to the volume of this aqueous solution and mix to separate into two layers, and collect the organic solvent layer such as ethyl acetate from there several times, preferably about three times. repeat. An organic fraction can be obtained by adding anhydrous sodium sulfate to the obtained organic solvent layer and then filtering it. After concentrating the organic fraction with an evaporator, the fraction eluted with hexane:ethyl acetate = 1:1 (v/v) etc. was eluted with a silica gel column chromatograph, and the fraction was eluted with a high performance liquid chromatograph using an ODS column (acetonitrile). : water = 60:40, 45:55 (both v/v)), the compound (gliasperin B) can be obtained.

The content of gliasperin B in the lipolysis promoter of the present invention is 0.00001% by mass to 50% by mass, preferably 0.0001% to 20% by mass, and 0.0005% to 10% by mass. %, and even more preferably 0.001% by mass to 5% by mass.

(Other ingredients)
In addition to gliasperin B, the lipolysis promoter of the present invention may contain carriers, excipients, solvents, and other optional components within a range that does not impair the effects of the present invention.

When the lipolysis promoter of the present invention contains gliasperin B, it may be blended with gliasperin B purified from licorice extract as described above or gliasperin B which is a chemically synthesized product, or finally Licorice extract may be blended so that the content of gliasperin B in the lipolysis promoter falls within the above numerical range. It is thought that the licorice extract may contain components that inhibit (suppress) the lipolytic action. Therefore, when the lipolysis promoter of the present invention contains gliasperin B, the licorice extract as described above is used. It is preferable that gliasperin B is blended with gliasperin B purified from natural substances or gliasperin B which is a chemically synthesized product. In addition, from the viewpoint of the lipolysis promoting effect, the lipolysis promoter of the present invention has a gliasperin B content within the above numerical range and does not contain licorice-derived components other than gliasperin B, or contains no licorice-derived components other than gliasperin B. When containing licorice-derived ingredients, use glycyrrhizic acid, which is a quantitative indicator of licorice extract in the Japanese Pharmacopoeia, as a comparison standard, and calculate the ratio of the content of gliasperin B to the content of glycyrrhizic acid (the content of gliasperin B to that of glycyrrhizic acid). The value divided by the content is preferably 0.001 or more.

The lipolysis promoter of the present invention can be used as a food or drink, a food with functional claims, a food for specified health use, a food with nutrient function claims, a cosmetic, a quasi-drug, a pharmaceutical product, a cosmetic, etc. by efficiently decomposing fat, subcutaneous fat, It is used to reduce body fat such as visceral fat.

When preparing the lipolysis accelerator of the present invention as a food or drink, in addition to Gliasperin B, sweeteners, coloring agents, preservatives, thickeners, stabilizers, gelling agents, thickening agents, antioxidants, and coloring agents are used. , bleach, fungicide, yeast food, gum base, fragrance, acidulant, seasoning, emulsifier, pH adjuster, kansui, leavening agent, nutritional fortifier, and other food and drink materials. , it may be prepared into a desired form. When the lipolysis promoter of the present invention is made into a food or drink form, the form is not particularly limited. Examples include supplement-type foods such as gels, granules, fine granules, capsules, tablets, powders, liquids, and semi-solids; carbonated drinks, soft drinks, milk drinks, alcoholic drinks, fruit juice drinks, teas, and nutritional drinks. Powdered beverages such as powdered juice and powdered soup; Confectionery such as gum, tablets, candies, cookies, gummies, rice crackers, biscuits, and jelly; Bread, noodles, cereals, jams, seasonings, etc. These foods are used as foods and beverages to efficiently break down fat and reduce body fat such as subcutaneous fat and visceral fat.For example, in addition to general foods and beverages, they are also used as nutritional supplements, foods with functional claims, It can also be used as a nutraceutical, such as food for specified health uses and food for the sick.

When preparing the lipolysis promoter of the present invention as a drug (including quasi-drugs), other medicinal ingredients, pharmaceutically acceptable carriers, additives, etc. may be added to Gliasperin B as necessary. They may be blended arbitrarily. Specific examples of pharmaceutically acceptable carriers and additives include binders, disintegrants, lubricants, wetting agents, buffers, preservatives, fragrances, and the like. When preparing the lipolysis promoter of the present invention as a pharmaceutical, there are no particular restrictions on its form. Examples include injections, external preparations, inhalants, suppositories, films, troches, liquids, powders, tablets, granules, capsules, syrups, eye drops, eyewashes, nasal drops, etc. can. Among these, forms suitable for oral administration (i.e. internal medicines) are preferred, and specific examples of such forms include troches, liquids, powders, tablets, granules, capsules, and syrups. . These medicines (including quasi-drugs) are used as medicines to efficiently decompose fat and reduce body fat such as subcutaneous fat and visceral fat.

To prepare the lipolysis promoter of the present invention as a cosmetic (including functional cosmetics) or a quasi-drug for external use, in addition to gliasperin B, a pharmaceutically or cosmetically acceptable carrier (water, Oily components, etc.) may be blended to prepare the desired form. The form of the above-mentioned cosmetic is not particularly limited as long as it can be applied to the skin. Examples include liquid, emulsion, powder, solid, suspension, cream, ointment, mousse, granule, tablet, gel, jelly, paste, gel, aerosol, and spray. , liniment agents, pack agents, and the like. These cosmetics are used as cosmetics for efficiently decomposing fat and reducing subcutaneous fat and the like.

The dosage and application amount of the lipolysis promoter of the present invention can be appropriately determined depending on the age, weight, health condition, degree of obesity, disease state, etc. of the person using the agent. In terms of total amount, 0.1 μg to 5000 μg, preferably 0.5 μg to 1000 μg, more preferably 1 μg to 500 μg, and even more preferably 5 μg to 100 μg are used.

The present invention will be explained in more detail with reference to Examples below, but the scope of the present invention is not limited thereto.

<Test 1> Preparation of Gliasperine B A licorice extract aqueous solution was prepared by adding and dissolving 10 ml of distilled water per 1 g of licorice extract. The operation of adding half the amount of ethyl acetate to the volume of this aqueous solution, mixing it, separating it into two layers, and recovering the ethyl acetate layer therefrom was repeated three times. Anhydrous sodium sulfate was added to the obtained ethyl acetate layer, and then filtered to obtain an organic fraction. After concentrating the organic fraction using an evaporator, the fraction eluted with hexane:ethyl acetate = 1:1 (v/v) was subjected to silica gel column chromatography and then subjected to high performance liquid chromatography using an ODS column (acetonitrile: A compound (Gliasperin B) was obtained by repeating purification using water = 60:40, 45:55 (both v/v). Gliasperin B (GB) obtained here was used as a test substance in the following test using mouse preadipocytes 3T3-L1.

The above-mentioned licorice extract was obtained by the following method. That is, the stem, root and stolone parts of licorice were ground to obtain licorice powder. The extract obtained by extracting 50 g of licorice powder with 500 mL of 50% water-containing ethanol at 20° C. was frozen and dried in an evaporator to obtain a 50% water-containing ethanol extract of licorice.

The spectral data of the compound is shown below, and it is very similar to the spectral data of gliasperin B, which has been reported to be isolated from plants of the genus Glycyrrhiza in the literature (Zeng L. et al., Heterocycles 34:575-587 (1992)). The compound was identified as gliasperin B (GB) because it showed good agreement with the protein and no contradiction with the structural information was observed.

1H-NMR (heavy acetone) ppm: 1.63(3H, s), 1.74(3H, s), 3.24(2H, d, J=7.0 Hz), 3.90(3H, s), 4.27(1H, dd, J= 5.5 and 10.5 Hz), 4.48(1H, dd, J=5.0 and 11.5 Hz), 4.62(1H, t, J=10.5 Hz), 5.17(1H, t, J=7.0 Hz), 6.13(1H, s) , 6.33(1H, dd, J=2.0 and 8.5 Hz), 6.45(1H, d, J=2.0 Hz), 6.94(1H, d, J=8.5Hz), 8.30(1H, bs), 8.63(1H, bs), 12.48(1H, s)
13C-NMR (heavy acetone) ppm: 17.6, 21.4, 25.7, 47.2, 56.2, 71.0, 91.3, 103.5, 103.7, 107.6, 109.7, 113.5, 123.3, 131.1,131.5, 156.8, 158.7 , 161.1, 162.7, 165.8, 198.7
ESI+-MS m/z: 393.13059[M+Na]+ (calculated value, C21H22O6Na: 393.13086)

Gliasperin B (GB) contained in the licorice extract was quantified by high performance liquid chromatography described in detail below. As a result, the content of gliasperin B (GB) was 132.6 μg per 1 g of licorice extract. Further, the glycyrrhizic acid contained in the above-mentioned licorice extract was 123 mg per 1 g of the licorice extract, and the value (ratio) of gliasperin B (GB) content/glycyrrhizic acid content was 0.00107.

(Analysis conditions for gliasperin B (GB))
Column; TSKgel ODS-100V 5μm (Tosoh) 4.6mm (inner diameter) x 150mm (length)
Column temperature: 40°C, mobile phase: acetonitrile:water = 45:55 (v/v)
Flow rate: 1ml/min
Detection wavelength: 290nm
GB retention time: 33.2 minutes

<Test 2> Examination of lipolytic effects of licorice extract and gliasperin B (GB) Mouse preadipocytes 3T3-L1 were cultured in differentiation induction medium (0.5mM Isobutyl-methylxanthine, 1μM Dexamethasone, 1μg/mL Insulin in 10% FBS) /DMEM) for 2 days, the medium was replaced with a 10% FBS/DMEM medium containing 1 μg/mL insulin, and cultured for 7 days. Test substances (licorice extract and GB) were added on day 9 from the start of differentiation induction, and then cultured for 19 hours. As the licorice extract, the licorice extract used in Test 1 was used (final concentration 100 μg/mL, 200 μg/mL, 300 μg/mL), and as GB, the GB prepared in Test 1 was used (final concentration 13. 5ng/mL, 27ng/mL, 40ng/mL). The test was conducted with 3 patients in each group. Glycerol released into the medium by decomposition of neutral fats accumulated in cells was quantified using Lab Assay (TM) triglyceride (Fuji Film Wako Pure Chemical Industries, Ltd.). The results are shown in Figure 1. The amount of glycerol was expressed as the average value ± standard deviation. Each symbol in FIG. 1 indicates the following. **p<0.01 vs. No treatment (Student's t-test).

As shown in Figure 1, the amount of glycerol released increases in a GB concentration-dependent manner, indicating that intracellular lipolysis occurs in a GB concentration-dependent manner, and GB has an effect of promoting lipolysis in adipocytes. It was found that it has On the other hand, glycerol was also released from adipocytes by licorice extract, but when the concentration of licorice extract was 300 μg/mL, the amount of glycerol released was lower than when the concentration was 100 μg/mL or 200 μg/mL. . Note that the respective concentrations of licorice extract, 100 μg/mL, 200 μg/mL, and 300 μg/mL, are converted into the amount of GB contained, respectively, to be 13.5 ng/mL, 27 ng/mL, and 40 ng/mL. Licorice extract also has an excellent lipolytic effect, but as its concentration increases, its lipolytic effect decreases. It was suggested that it may contain inhibiting ingredients. From this, it can be said that in order to obtain a higher lipolytic effect, it is preferable to use a purified product of GB rather than a licorice extract.

<Test 3> Effect of licorice extract on body weight, BMI, and visceral fat level (indicator value of visceral fat accumulation) A subject (one 59-year-old male) received 200 mg of the licorice extract used in Study 1 per day. , was ingested for 100 days. Subjects were given a recording sheet, and their weight, BMI, and visceral fat level were measured using a Tanita body composition analyzer before the start of licorice extract intake (day 0), on the 30th day of intake, on the 45th day of intake, and on the 72nd day of intake. Recordings were made on the 83rd day of intake, the 92nd day of intake, and the day after the last intake (101st day). Figure 2 shows the changes in body weight, BMI, and visceral fat level from before the start of intake.

As shown in FIG. 2, it was shown that ingestion of GB-containing licorice extract led to a decrease in visceral fat level, followed by a decrease in body weight and BMI. Considering the results of Test 2 above, it was considered that GB in the licorice extract efficiently decomposed fat in adipocytes and reduced body fat such as visceral fat.

The lipolysis promoter of the present invention exhibits an excellent lipolysis promoting effect by containing gliasperin B as an active ingredient. By using the lipolysis promoter of the present invention, fat in fat cells can be efficiently decomposed and body fat such as subcutaneous fat and visceral fat can be reduced. Therefore, according to the lipolysis promoter of the present invention, obesity and lifestyle-related diseases can be prevented and improved over the long term, and health can be maintained and improved.

Claims (8)

A lipolysis promoter containing gliasperin B as an active ingredient. The lipolysis promoter according to claim 1, wherein the gliasperin B is derived from a licorice extract. The lipolysis promoter according to claim 2, wherein the licorice extract is a hydrous ethanol extract of licorice. The lipolysis promoter according to claim 3, wherein the ethanol concentration in the water-containing ethanol is 0.1% (v/v) to 99.9% (v/v). The lipolysis promoter according to claim 4, wherein the ethanol concentration in the water-containing ethanol is 30% (v/v) to 70% (v/v). The lipolysis promoter according to any one of claims 1 to 5, wherein the content of gliasperin B is 0.00001% by mass to 50% by mass. The lipolysis promoter according to any one of claims 1 to 6, which is administered in an amount of gliasperin B of 5 μg to 100 μg per day. According to any one of claims 1 to 7, the product does not contain glycyrrhizic acid, or when it contains glycyrrhizic acid, the value obtained by dividing the content of gliasperin B by the content of glycyrrhizic acid is 0.001 or more. The lipolysis promoter described.