Internal composition for lipolysis and / or weight loss, method for determining effectiveness thereof, and quality index for plant extract

Abstract

Provided is a novel preparation having an excellent lipolysis-promoting effect, an excellent visceral fat and body fat reducing effect, an excellent body weight reducing effect, and an excellent BMI improving effect. Another purpose of the present invention is to provide a novel preparation having an excellent insulin resistance improving effect, an excellent cholesterol value reducing effect, and the like. The present invention is an internal composition containing hirsutin B, which is used for at least one of lipolysis, fat burning, fat consumption assistance, BMI improvement, BMI reduction assistance, abdominal fat reduction, visceral fat reduction, body fat reduction, waist reduction, insulin resistance improvement, and LDL cholesterol value reduction. The internal composition of the present invention is preferably used for adults having a BMI of 23 kg / m < 2 > or more and less than 30 kg / m < 2 > and for adults having a visceral fat area of 100 cm < 2 > or more.

Description

TECHNICAL FIELD

[0001] The present application relates to an internal composition for fat decomposition and / or weight reduction, a method for determining effectiveness, and a quality indicator for a plant extract. BACKGROUND

[0002] Body fat is generated when the excess portion of the energy intake over the energy consumption is accumulated as neutral fat in white adipocytes. Excessive fat accumulation leads to obesity and weight gain, and as a result, such obesity not only becomes a cause of various lifestyle-related diseases, but also constitutes a major problem in terms of beauty. Obesity in which visceral fat is mainly accumulated in body fat has been pointed out to be associated with pathological conditions such as insulin resistance or arteriosclerosis, and in addition, obesity in which subcutaneous fat is mainly accumulated has become a matter of high concern from the viewpoint of beauty, regardless of men or women.

[0003] In the past, various measures for limiting energy intake, such as dietary control, searching for substances that hinder sugar absorption in the digestive tract, and the like, have been taken for the prevention and improvement of obesity. However, limiting energy intake can lead to a decrease in basal metabolic rate, and sometimes cannot improve obesity. In addition, energy intake limitation can also lead to a deficiency of essential nutrients, thereby impairing health. Therefore, in order to eliminate obesity, it is currently widely believed that an ideal solution should be to actively decompose accumulated fat and release it as heat energy, thereby being able to maintain an ideal body weight. In this context, in recent years, the exploration of functional ingredients having a fat decomposition-promoting effect in food raw materials has been increasingly active, and many food and beverages have been proposed.

[0004] For example, it is known that a mixture prepared by combining salmon white extract, beer yeast extract, barley leaflet essence, and type III collagen (chicken collagen) has an excellent fat decomposition-promoting effect (Patent Literature 1), at least either one of birch extract of Betulaceae and extract of Carex spicata of Gramineae improves obesity constitution by promoting fat decomposition in whole body or local adipose tissue, and prevents adipose tissue enlargement (Patent Literature 2), a specific plant or extract thereof such as European juniper promotes decomposition of accumulated neutral fat in adipose tissue, and is useful as a medicine, food, or cosmetic that exerts a slimming effect for inhibition, prevention, or improvement of obesity and the like (Patent Literature 3), and the like. However, the fat decomposition effect, weight reduction effect, and the like of these plant extracts and the like are not sufficient at present. In addition, it is disclosed in Patent Literature 4 that a composition in which a glycyrrhiza hydrophobic extract is combined with an antioxidant ingredient and the like exerts effects of inhibiting body fat accumulation, promoting body fat decomposition, and promoting energy production, but a sufficient fat decomposition-promoting effect cannot be obtained by the glycyrrhiza hydrophobic extract alone.

[0005] PRIOR ART DOCUMENTS

[0006] PATENT LITERATURE

[0007] [Patent Literature 1] Japanese Patent Laid-Open No. 2011-074051

[0008] [Patent Literature 2] Japanese Patent Laid-Open No. 2006-045120

[0009] [Patent Literature 3] Japanese Patent Laid-Open No. 2012-229266

[0010] [Patent Literature 4] International Publication No. 2008 / 143182 SUMMARY

[0011] PROBLEMS TO BE SOLVED BY THE INVENTION

[0012] The present application aims to provide a novel preparation which is excellent in fat decomposition promotion effect, visceral fat and body fat reduction effect, body weight reduction effect, BMI (Body Mass Index) improvement effect, and the like. Further, the present application aims to provide a novel preparation which is excellent in insulin resistance improvement effect, cholesterol value reduction effect, and the like.

[0013] METHOD FOR SOLVING THE PROBLEM

[0014] The present inventors have conducted intensive studies, and as a result, have found that "Kurorin B", which is a specific component in liquorice extract, has an excellent fat decomposition promotion effect. Further, it has been found that a preparation containing liquorice extract containing Kurorin B has visceral fat and body fat reduction effect, body weight reduction effect, BMI improvement effect, insulin resistance improvement effect, cholesterol value reduction effect, in addition to the excellent fat decomposition promotion effect. Furthermore, it has been found that the content of Kurorin B in a plant extract becomes an index of the effectiveness of the plant extract on fat decomposition, body weight reduction, and the like. The gist of the present application is as follows.

[0015] [1] An internal use composition containing Kurorin B for use in any one or more of fat decomposition, fat combustion, fat consumption aid, BMI improvement, BMI reduction aid, abdominal fat reduction, visceral fat reduction, body fat reduction, waist circumference reduction, insulin resistance improvement, and LDL cholesterol value reduction.

[0016] [2] An internal use composition containing an effective amount of Kurorin B for use in any one or more of fat decomposition, fat combustion, fat consumption aid, BMI improvement, BMI reduction aid, abdominal fat reduction, visceral fat reduction, body fat reduction, waist circumference reduction, insulin resistance improvement, and LDL cholesterol value reduction.

[0017] [3] The internal use composition according to [2], wherein the effective amount is 10 μg or more per day for an adult.

[0018] [4] The composition for internal use according to [1] or [2], which is used for an adult whose BMI is 23 kg / m 2 or less and less than 30 kg / m 2 .

[0019] [5] The composition for internal use according to [1] or [2], which is used for an adult whose visceral fat area is 100 cm 2 or more.

[0020] [6] The composition for internal use according to [1] or [2], wherein the intake amount is 5 to 100 μg / day in terms of the total amount of picrocrocin B.

[0021] [7] A method for determining effectiveness, which determines the effectiveness of a plant extract on fat decomposition and / or weight reduction in a living organism, using the content of picrocrocin B as an index.

[0022] [8] The method for determining the effectiveness on fat decomposition and / or weight reduction according to [7], wherein the plant is Glycyrrhiza.

[0023] [9] An index which is a quality index of a plant extract, characterized in that the quality is a quality related to the effectiveness on fat decomposition and / or weight reduction in a living organism, and is constituted by picrocrocin B.

[0024]

[10] The index according to [9], wherein the plant is Glycyrrhiza.

[0025] Effects of the Invention

[0026] The composition for internal use of the present invention has an excellent fat decomposition promoting effect by containing picrocrocin B, preferably by containing it as an effective component, a functionally participating component, an index component, or a contributing component, and exerts a remarkable weight reduction effect. By using the composition for internal use of the present invention, fat in adipocytes can be efficiently decomposed, and body fat such as subcutaneous fat and visceral fat can be reduced, and finally a remarkable weight reduction effect and a BMI improvement effect can be obtained. In addition, an effect of improving insulin resistance and lowering cholesterol values can be obtained. Therefore, the composition for internal use according to the present invention can prevent and improve obesity or lifestyle-related diseases for a long period of time, and can realize the maintenance and enhancement of health. Furthermore, since the content of picrocrocin B in a plant extract becomes an index of the effectiveness of the plant extract on fat decomposition, weight reduction, and the like, a plant extract having high effectiveness can be efficiently selected. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 is a graph showing the fat decomposition promoting effect of Glycyrrhiza extract and picrocrocin B.

[0028] Figure 2 is a graph showing the effect of licorice extract on body weight, BMI, and visceral fat level. DETAILED DESCRIPTION

[0029] Hereinafter, the internal composition of the present application, the method for determining the effectiveness on fat decomposition and / or weight reduction, and the quality index of plant extract are described in detail.

[0030] <Internal composition>

[0031] The internal composition of the present application contains glypiperide B. Although not limited, it is preferable to contain glypiperide B as an effective component, a functionally participating component, an index component, or a contributing component. The internal composition of the present application can contain other components in addition to glypiperide B as a necessary component, within a range not impairing the effect of the present application. Hereinafter, glypiperide B and other components contained in the internal composition of the present application, the form of the internal composition of the present application, and the like are described.

[0032] In the present specification, the effective component refers to a pharmacologically active component or a physiologically active component that exhibits its target effect among components contained in a pharmaceutical product, a quasi-pharmaceutical product, or the like.

[0033] In the present specification, the functionally participating component refers to a component that contributes to a specific health purpose among components contained in an internal composition such as a food product. Specifically, it refers to a component for which the mechanism of action related to functionality has been investigated through in vitro tests and in vivo tests or clinical tests (human tests), and for which direct or indirect qualitative confirmation and quantitative confirmation are possible. In addition, a functional claim food is a product that claims a function based on the effect of the components formulated in the functional claim food and provides it to a demander, and the content of the above-mentioned functionally participating component or a method for quantifying it must be reported when it is provided to the demander as a functional claim food.

[0034] In the present specification, the index component refers to a specific component that is known to explain part of the scientific basis of functionality, but in the case of an extract, an essence, or the like as a functionally participating component, which cannot explain the entire functionality only by the specific component, there is at least one component that is investigated through in vitro tests and in vivo tests or clinical tests (human tests) for the mechanism of action related to the functionality to be claimed, and such a component is the index component.

[0035] In the present specification, a contributing component refers to a substance, and a component that directly or indirectly affects the physiological function of the body by being formulated. In specific health foods and functional foods, "participating components", "functionally participating components", "index components", and the like are considered as components equivalent to contributing components, but are not limited to these. Furthermore, in pharmaceuticals and quasi-pharmaceuticals, "effective components" are equivalent to contributing components. Furthermore, the concept also covers components that are impliedly associated with effects and emphasized in the form of "formulated ○○ component. Assist in burning" and the like in the process of selling food compositions other than functional foods.

[0036] (Glyasperin B)

[0037] Glyasperin B (hereinafter also referred to as "GB") is an isoflavone derivative represented by the following formula. Also represented as 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-(2,4-dihydroxyphenyl)-5-hydroxy-7-methoxy-6-(3-methylbut-2-en-1-yl)-3,4-dihydro-2H-1-benzopyran-4-one.

[0038] [Chemical 1]

[0039]

[0040] Glyasperin B is a component that has been confirmed to be contained in licorice extract, and as Glyasperin B in the present application, it is preferable to be derived from licorice extract. So far, it has been known that licorice extract exerts a fat decomposition-promoting effect, and there are documents suggesting that glabridin, which is contained in a large amount in licorice extract, can exert a fat decomposition-promoting effect (see Patent Literature 4 and the like). However, it has not been clear which component in licorice extract exerts a fat decomposition-promoting effect. The present inventors and the like have first found that, among components that are present in a large amount in licorice extract, Glyasperin B exerts a fat decomposition-promoting effect. Furthermore, the present inventors and the like have completed the invention of an internal composition containing this Glyasperin B and exerting effects of fat decomposition, fat burning, fat consumption assistance, BMI improvement, BMI reduction assistance, abdominal fat reduction, visceral fat reduction, body fat reduction, waist circumference reduction, improvement of insulin resistance, and LDL cholesterol value reduction.

[0041] Glyasperin B in the present application can be derived from licorice, or can be a chemical synthetic product. In addition, in the present specification, the case of containing Glyasperin B includes the case of containing licorice and the like plants containing Glyasperin B.

[0042] The above-mentioned licorice species are not particularly limited, and examples of the licorice species include plants of the genus Glycyrrhiza of the family Leguminosae, such as G. uralensis Fisch. et DC, G. inflata BAT., G. glabra L., G. glabra L. var. glandulifera Regel et Herder, G. aspera, G. echinata L., G. pallidiflora Maxim, and the like.

[0043] As a method for obtaining crude asperosaponin B derived from licorice, a method of purification from a licorice extract can be exemplified. The licorice extract, in particular, refers to an extract obtained by extracting a pulverized product of the whole plant or a part (e.g., roots, stems, stolons, leaves, flowers, fruits, and the like) of licorice with a solvent, and can also be an extract obtained by spray drying or freeze drying the obtained licorice extract. As the extraction solvent, water, or an alcohol such as methanol or ethanol, or a mixed solvent of water and an alcohol or a ketone such as acetone, and the like can be exemplified. Among these, water, an alcohol, an aqueous alcohol, hot water, ethanol, and an aqueous ethanol are preferable as the extraction solvent.

[0044] The alcohol concentration of the above-mentioned aqueous alcohol is 0.1% to 99.9% by mass, preferably 10% to 99.9% by mass, more preferably 30% to 70% by mass, further preferably 40% to 60% by mass, and 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), more preferably 30% (v / v) to 70% (v / v), further preferably 40% (v / v) to 60% (v / v), and particularly preferably 50% (v / v).

[0045] A dried product of the above-mentioned licorice extract is dissolved by adding distilled water thereto, and a licorice extract aqueous solution is prepared. An organic solvent such as ethyl acetate is added to half the volume of the aqueous solution, and mixed to become a state in which two layers are separated, and the organic solvent layer such as ethyl acetate is recovered therefrom and the operation is repeated several times, preferably about three times. Anhydrous sodium sulfate is added to the obtained organic solvent layer, and an organic fraction is obtained by filtration. The organic fraction is concentrated with an evaporator, and eluted with a silica gel column chromatography method using hexane: ethyl acetate = 1: 1 (v / v) or the like, and the obtained fraction is repeatedly purified by a high-performance liquid chromatograph using an ODS column (acetonitrile: water = 60:40, 45:55 (both v / v)) to obtain a compound (crude asperosaponin B).

[0046] The content of pungaricin B in the internal composition of the present application is 0.00001 to 50 mass%, preferably 0.0001 to 20 mass%, more preferably 0.0005 to 10 mass%, and further preferably 0.001 to 5 mass%.

[0047] (Other components)

[0048] The internal composition of the present application can contain, in addition to pungaricin B, carriers, excipients, solvents, and other arbitrary components, without impairing the effects of the present application.

[0049] The internal composition of the present application, when containing pungaricin B, can be formulated using pungaricin B purified from the glycyrrhiza extract as described above, or pungaricin B as a chemical synthetic product, or can be formulated so that the content of pungaricin B in the final internal composition becomes the above numerical range, using the glycyrrhiza extract. In addition, from the viewpoint of the lipolysis-promoting action, in the internal composition of the present application, when the content of pungaricin B is within the above numerical range, and the internal composition does not contain components derived from glycyrrhiza other than pungaricin B, or contains components derived from glycyrrhiza other than pungaricin B, the ratio of the content of pungaricin B to the content of glycyrrhetic acid (the value obtained by dividing the content of pungaricin B by the content of glycyrrhetic acid) is preferably 0.001 or more, using the quantitative index of glycyrrhetic acid in the Japanese Pharmacopoeia as a comparative reference.

[0050] The internal composition of the present application is used for efficiently decomposing fat, reducing subcutaneous fat, visceral fat, and the like, as a food and beverage, a functional labeled food, a specified health food, a nutritionally functional food, a cosmetic, a quasi-drug, a pharmaceutical, or a cosmetic, and the like.

[0051] When the internal use composition of the present application is prepared as a food or beverage, in addition to glyyrrhizin B, a sweetening agent, a coloring agent, a preservative, a viscosity increasing agent, a stabilizer, a gelling agent, a pasting agent, an antioxidant, a color developing agent, a bleaching agent, a mold preventing agent, a yeast food, a gum base, a flavoring agent, a souring agent, a seasoning agent, an emulsifying agent, a pH adjusting agent, an alkali water, a leavening agent, a nutritional supplement, other food or beverage raw materials, and the like can be mixed to prepare a desired form. When the internal use composition of the present application is prepared as a food or beverage, the form is not particularly limited. As an example, a supplement food such as a gel-like agent, a granule, a fine granule, a capsule, a tablet, a powder, a liquid agent, a semi-solid agent, a beverage such as a carbonated beverage, a refreshing beverage, a milk beverage, an alcoholic beverage, a juice beverage, a tea, a nutritional beverage, a powdered beverage such as a powdered juice, a powdered soup, a confectionery such as chewing gum, a lozenge, a candy, a cookie, a gummy candy, a senbei, a biscuit, a jelly, a bread, a noodle, a cereal, a jam, a seasoning, and the like can be exemplified. These foods and beverages are used as a food or beverage for efficiently decomposing fat and reducing body fat such as subcutaneous fat and visceral fat.

[0052] When the internal use composition of the present application is prepared as a pharmaceutical product (including a quasi-drug), in addition to glyyrrhizin B, other pharmaceutical ingredients, a pharmaceutically acceptable carrier or additive, and the like can be arbitrarily formulated as needed. As the pharmaceutically acceptable carrier and additive, specifically, a binding agent, a disintegrating agent, a lubricant, a wetting agent, a buffer, a preservative, a flavoring agent, and the like can be exemplified. When the internal use composition of the present application is prepared as a pharmaceutical product, the form is not particularly limited. As an example, an injection, a topical agent, an inhalant, a suppository, a film, a lozenge, a liquid agent, a powder, a tablet, a granule, a capsule, a syrup, an eye drop, an eye wash, a nose drop, and the like can be exemplified. Among them, a form suitable for oral administration (i.e., an internal use pharmaceutical product) is preferred, and as the form, specifically, a lozenge, a liquid agent, a powder, a tablet, a granule, a capsule, a syrup, and the like can be exemplified. These pharmaceutical products (including quasi-drugs) are used as a pharmaceutical product for efficiently decomposing fat and reducing body fat such as subcutaneous fat and visceral fat.

[0053] The picrorhizin B contained in the internal composition of the present application can also be used as a cosmetic product (including a functional cosmetic product) or an external medical product. When prepared as a cosmetic product or an external medical product, a pharmaceutically or cosmetically acceptable carrier (water, an oily ingredient, etc.) is formulated in addition to the picrorhizin B, and the desired form is prepared. The form of the cosmetic product is not particularly limited as long as it is applicable to the skin. As an example, the following forms can be listed: a liquid form, an emulsion form, a powder form, a solid form, a suspension form, a cream form, an ointment form, a mousse form, a granule form, a tablet form, a gel form, a jelly form, a paste form, a gum form, an aerosol form, a spray form, a tincture, a dressing agent, and the like. These cosmetic products are used as cosmetic products for efficiently decomposing fat, reducing subcutaneous fat, and the like.

[0054] The amount and the appropriate amount of the internal composition of the present application can be appropriately determined according to the age, the body weight, the health condition, the obesity degree, the disease condition, and the like of the user, and are not limited, and the total amount of the picrorhizin B can be set to 0.1 μg or more, 0.5 μg or more, 1 μg or more, 5 μg or more, 10 μg or more, 12 μg or more, and can be set to 5000 μg or less, 1000 μg or less, 500 μg or less, 100 μg or less, 60 μg or less, 20 μg or less, 15 μg or less, and the like, per day for an adult. For example, the intake amount of the picrorhizin B is 0.1 μg to 5000 μg, preferably 0.5 μg to 1000 μg, more preferably 1 μg to 500 μg, further preferably 1 μg to 200 μg, furthermore preferably 1 μg to 100 μg, particularly preferably 5 μg to 100 μg, particularly more preferably 5 μg to 60 μg, particularly further preferably 10 μg to 50 μg, and most preferably 10 μg to 20 μg, per day for an adult.

[0055] When the internal use composition of the present application is administered orally to a human, an effective amount of the anemoside B is preferably administered. In the present specification, the effective amount means an amount of administration at which the target effect can be seen, whether directly or indirectly. As the effective amount, it differs depending on the age or the body weight, and for example, it can be set to 1 μg or more, 3 μg or more, 5 μg or more, 7 μg or more, 10 μg or more, 12 μg or more, and 1 mg or less, 500 μg or less, 200 μg or less, 100 μg or less, 60 μg or less, 50 μg or less, 20 μg or less, etc. in terms of the total amount of the anemoside B per day for an adult. For example, it is 1 μg or more and 1 mg or less, preferably 1 μg or more and 500 μg or less, more preferably 1 μg or more and 200 μg or less, further preferably 1 μg or more and 100 μg or less, further more preferably 5 μg or more and 100 μg or less, particularly preferably 5 μg or more and 60 μg or less, particularly more preferably 10 μg or more and 50 μg or less, particularly further preferably 10 μg or more and 20 μg or less, and most preferably 14 μg in terms of the total amount of the anemoside B per day for an adult.

[0056] From the viewpoint of easily obtaining the weight reduction effect, the internal use composition of the present application is preferably used for an adult whose BMI is 23 kg / m 2 or more and less than 30 kg / m 2 . More preferably, it is used for an adult whose BMI is 23 kg / m 2 or more and less than 25 kg / m 2 . According to the criteria of the Japan Society for the Obesity, the BMI of 18.5 kg / m 2 or more and less than 25 kg / m 2 is set to normal weight, and the BMI of 25 kg / m 2 or more and less than 30 kg / m 2 is set to obesity degree 1. The internal use composition of the present application is preferably used for an adult of the so-called normal weight to the obesity degree 1, and particularly preferably for an adult who is not classified as obese but is in the normal weight to the obesity degree 1. In addition, the BMI (Body Mass Index) is also called the body mass index, and is a physical index indicating the obesity degree calculated from the body weight and the height by the following calculation formula. In an adult, the BMI is used as an international index.

[0057]

[0058] From the viewpoint of easily obtaining the fat decomposition promoting effect and the weight reduction effect, the internal use composition of the present application is preferably used for an adult whose visceral fat area is 100 cm 2 or more. The visceral fat area can be calculated by performing abdominal CT scanning to take a plurality of slice images at 1 cm intervals, etc. from the navel as the center, and according to the conventional method.

[0059] Another embodiment of the present application is a weight loss agent, visceral fat, body fat loss agent, BMI improver, insulin resistance improver, or cholesterol value reducer containing glycyrrhizin B as an effective ingredient.

[0060] The present application also includes the above-mentioned weight loss agent, visceral fat, body fat loss agent, BMI improver, insulin resistance improver, or cholesterol value reducer, which has an excellent fat decomposition promoting effect by containing glycyrrhizin B, and exerts a remarkable weight loss effect. By using the weight loss agent, visceral fat, body fat loss agent, BMI improver, insulin resistance improver, or cholesterol value reducer of the present application, fat in fat cells can be efficiently decomposed, and subcutaneous fat, visceral fat, and the like can be reduced, and finally a remarkable weight loss effect, BMI improvement effect can be obtained. In addition, an effect of improving insulin resistance and reducing cholesterol value can also be obtained. Therefore, the weight loss agent, visceral fat, body fat loss agent, BMI improver, insulin resistance improver, or cholesterol value reducer of the present application can prevent and improve obesity or lifestyle-related diseases for a long period of time, and can achieve maintenance and improvement of health. In addition, the weight loss agent, visceral fat, body fat loss agent, BMI improver, insulin resistance improver, or cholesterol value reducer of the present application has the same constitution as the above-mentioned internal composition of the present application containing glycyrrhizin B as an effective ingredient, and thus the specific description can be directly applied to the description of the internal composition of the present application.

[0061] <Method for determining effectiveness of plant extract on fat decomposition and / or weight loss>

[0062] The present application also includes a method for determining effectiveness of a plant extract on fat decomposition and / or weight loss in an organism using the content of glycyrrhizin B as an index. As described above, a preparation containing glycyrrhizin B as an effective ingredient has an excellent fat decomposition promoting effect, and exerts a remarkable weight loss effect. In addition, fat in fat cells can be efficiently decomposed, and subcutaneous fat, visceral fat, and the like can be reduced, and finally a remarkable weight loss effect, BMI improvement effect can be obtained. In addition, an effect of improving insulin resistance and reducing cholesterol value can also be obtained. The above-mentioned effects are also enhanced in accordance with the content of glycyrrhizin B, and thus the effectiveness of a plant extract on fat decomposition and / or weight loss in an organism can be determined by using the content of glycyrrhizin B in the plant extract as an index. As the above-mentioned plant extract, there is no particular limitation as long as it contains glycyrrhizin B, and a licorice extract is preferred.

[0063] The determination method of the present application will be described in detail below. The determination method of the present application includes a step of measuring the content of tirucallol B in a plant extract (hereinafter, also referred to as "the measuring step"). In addition, it is preferable to include a step of comparing the amount of tirucallol B obtained with the amount of tirucallol B as a reference (hereinafter, also referred to as "the comparing step").

[0064] In the measuring step, the method of measuring the content of tirucallol B in a plant extract is not particularly limited, and the method known to those skilled in the art can be used. For example, the following methods can be mentioned: a method of performing Western blotting, a method using ELISA (Enzyme Linked Immunosorbent Assay), and the like. In either the method of performing Western blotting or the method using ELISA, an anti-tirucallol B antibody that specifically binds to tirucallol B can be used to measure the content of tirucallol B with high accuracy. In the method of performing Western blotting, the content of tirucallol B in a sample can be calculated by comparing the intensity of the obtained tirucallol B-specific band with the intensity of a standard (positive control) of known concentration.

[0065] Tirucallol B used as a reference in the determination method of the present application can be obtained, for example, by extraction from a natural source, expression of a recombinant nucleic acid encoding tirucallol B, or chemical synthesis.

[0066] In the determination method of the present application, the content of tirucallol B in the extract to be evaluated is compared with the content of tirucallol B as a reference by the comparing step. The amount of tirucallol B as a reference can be appropriately selected depending on the purpose and object of the evaluation.

[0067] <Quality indicator of plant extract>

[0068] The present application also includes an indicator, which is a quality indicator of a plant extract, characterized in that the quality is a quality related to the effectiveness of fat decomposition and / or body weight reduction in an organism, and is constituted by tirucallol B. As described above, a preparation containing tirucallol B as an effective ingredient has an excellent fat decomposition-promoting effect, and exerts a remarkable body weight reduction effect. In addition, it can efficiently decompose fat in fat cells, reduce body fat such as subcutaneous fat and visceral fat, and finally exerts a remarkable body weight reduction effect and a BMI improvement effect. In addition, it can also exerts an effect of improving insulin resistance and lowering cholesterol values. Depending on the content of tirucallol B, the above effects are also enhanced accordingly, and thus by using tirucallol B in a plant extract as an indicator, the effectiveness of the plant extract on fat decomposition and / or body weight reduction in an organism can be determined. As the plant in the plant extract, licorice is preferable.

[0069] The quality index of the plant extract of the present application corresponds to the pungarol B in the above-mentioned method for determining the effectiveness of the plant extract on fat decomposition and / or weight reduction. The specific description of the index of the present application can apply the description in the above-mentioned determination method.

[0070] [Examples]

[0071] The following examples more specifically illustrate the present application, but the scope of the present application is not limited to these examples.

[0072] As shown below, in order to evaluate the effect on visceral fat after 12 weeks of intake of the test food (hard capsule preparation formulated with licorice extract) by the subjects, a randomized double-blind parallel group comparison test was conducted with the control food (hard capsule not formulated with licorice extract) as a control.

[0073] [Example 1]

[0074] 1. Test food (composition, packaging, etc.)

[0075] The test food contains 100 mg of licorice extract (manufactured by MG Pharma (Co.) derived from Glycyrrhiza uralensis) per one piece and is composed of other excipients. The control food does not contain licorice extract and is composed of excipients colored with caramel pigment. The shape of the test food is a white hard capsule, and 30 pieces are packaged in an aluminum bag with an identification code marked on the test food. The unrecognizability of the test food and the package has been reviewed and approved by the IRB (institutional review board). The nutritional components of the test food and the participation components are shown in Table 1 below. In addition, the amount of pungarol B contained in the test food is 14 μg of the standard amount of daily intake.

[0076] [Table 1]

[0077]

[0078] 2. Subjects

[0079] As for the number of target medical records in this test, in the parallel group comparison test, assuming that the effect size of the test food is large (0.8), the significance level is set to 0.05, and the test power is set to 80-90%, 26-35 people are needed in each group, so the target number of medical records is calculated to be 34 people in each group, a total of 68 people. In addition, considering the dropout and test termination in the test, the number of applicants is set to 80 people (40 people in each group). The subjects were publicly recruited from volunteers registered with the test commissioning agency, and 81 people (50 men and 31 women) met the following selection criteria and did not violate the exclusion criteria, and were judged by the test physician to be suitable for participating in this test, and were included as subjects.

[0080] (Selection Criteria)

[0081] People who meet the following conditions in the screening examination.

[0082] (1) Men and women over 20 years old and under 65 years old, (2) BMI of 23.0 kg / m 2 or less and under 30.0 kg / m 2 , (3) Visceral fat area of 100 cm 2 or more, (4) People who have been fully informed of the purpose and content of the test, have the ability to consent, and voluntarily apply on the basis of full understanding and written consent to participate in the test.

[0083] (Exclusion Criteria)

[0084] (1) those with severe cardiovascular disease, liver dysfunction, renal dysfunction, respiratory disease, endocrine disease, metabolic disease, or with a relevant past medical history; (2) those receiving treatment for chronic diseases such as dyslipidemia, hypertension, diabetes, etc.; (3) those with mental illness such as depression, schizophrenia, anorexia nervosa, etc.; (4) those who frequently use pharmaceuticals, quasi-drugs, which can affect the evaluation of body fat, neutral fat, body weight, cholesterol, energy metabolism, etc. in this test; (5) those who frequently use supplements, health foods (special health foods, functional foods, etc.), which can affect the evaluation of body fat, neutral fat, body weight, cholesterol, energy metabolism, etc. in this test; (6) those who can have an allergy to the test food; (7) those who have metal in the abdominal CT scan site; (8) those with implanted medical devices such as cardiac pacemakers, implantable defibrillators, etc.; (9) those with claustrophobia; (10) those with a smoking habit; (11) those with a heavy drinking habit (60 g or more of pure alcohol per day); (12) those with extremely irregular eating habits; (13) those working night shifts and shift work; (14) those who have not defecated for 5 days or more; (15) those with menopausal symptoms; (16) those with hypertension, pseudohyperaldosteronism, hypokalemia, myopathy; (17) those who frequently eat medical foods with licorice and foods mainly containing licorice; (18) those who are prone to edema and fatigue; (19) those with urinary dysfunction; (20) those who have had blood drawn (blood donation, etc.) of 200 mL or more within 1 month before the start of the test or 400 mL or more within 3 months; (21) those who have participated in other clinical tests within the past 4 months, those who are currently participating in other clinical tests, those who plan to participate in other clinical tests during the test period; (22) pregnant, lactating, or planning to become pregnant women; (23) those judged by the test director to be unsuitable for participation.

[0085] 3. Test protocol

[0086] This test is a randomized placebo-controlled double-blind parallel group comparison test. Examinations are conducted at the start of test food intake, after 4 weeks of intake, after 8 weeks of intake, and after 12 weeks of intake. For subjects included by the test director, a randomized random number is generated by the allocation director who does not participate in the test implementation and is allocated to the subjects. At this time, there are no significant differences in the components among the gender at the time of consent, age, visceral fat area, BMI, hip circumference, waist circumference at the time of screening examination, as allocation factors.

[0087] During the test period, one capsule of test food (test food group: test food, control food group: control food) is taken once a day, and is taken with water or warm water before dinner. However, if it cannot be taken before dinner, one capsule is taken once a day at the earliest opportunity.

[0088] During the test period, the diet should be maintained as much as possible in the normal habit, and the intake of health foods (specific health foods, functional foods, etc.), and grapefruit juice, which can affect the evaluation of body fat, neutral fat, body weight, cholesterol, energy metabolism, etc. of this test, should be prohibited. In addition, the amount of alcohol intake should be maintained as the normal intake habit, and the drinking method should be prohibited from deviating from the usual intake amount. In addition, alcohol should be prohibited from being ingested on the day before the examination, and anything other than water should be prohibited from being ingested after 21 o'clock. In addition, anything other than water should be prohibited from being ingested for 4 hours before the abdominal CT scan. During the test period, the normal exercise habit (frequency, type of exercise, etc.) should be maintained as much as possible, and the exercise habit should not be changed. During the test period, the use of new medicines should be permitted with the permission of the doctor in charge of the test, except in emergency situations. Vaccination should be prohibited 2 weeks before the examination day. Blood donation should be prohibited during the test period.

[0089] This test was conducted in accordance with the spirit of the Declaration of Helsinki (2013) - Tokyo, Venice, Hong Kong, West Somerset, Edinburgh revision, Washington, Tokyo annotation supplement, Seoul revision, Fortaleza revision, in accordance with "Ethical Guidelines for Life Science and Medical Research on Human Subjects (Designated on March 23, 2021, Implemented on June 30, 2021, Partially revised on March 10, 2022)". It was approved by the Medical Corporation Kosei Clinic Fukuda Internal Medicine Clinic Ethics Review Committee (IRB) at the time of implementation (Approval No.: IRB-20220716-2). In addition, clinical trial registration has been conducted in the University Hospital Medical Information Network Clinical Trial Registry (UMIN) (UMIN Trial ID: UMIN000048430).

[0090] 4. Examination items

[0091] 1) Physical examination and abdominal CT scan

[0092] The items of physical examination are measurement of height (only at the time of screening examination), body composition evaluation using InBody770 (InBody Japan (stock)), waist circumference, hip circumference, blood pressure, and pulse, and CT scan for those who meet the selection criteria. Regarding body weight, DST-210N (Muratec KDS (stock)) is used only at the time of CT scan. In addition, BMI is calculated from height and weight, and the BMI at the time of CT scan is used as the value before the start of intake.

[0093] Abdominal CT scan using Discovery 710 or Discovery MIDR (GE Healthcare Japan (K.K.)) to take three slice images at 1 cm intervals centered on the umbilical region, and calculate the average of visceral fat area, subcutaneous fat area, and total fat area. In addition, if the image at the time of scanning the umbilical region includes the kidney and ilium, re-scan centered on the fourth lumbar vertebra.

[0094] 2) Diet and activity records

[0095] During the test period, the subjects were asked to record alcohol intake, medication use, diet content, etc. in a life record log. During the test food intake period, the intake of test food was recorded. In addition, three days before the examination, in addition to recording the diet content, the amount of food was also recorded. During the test period, in addition to the bedtime, the subjects were asked to wear an activity meter (SAT-1, Medisync (K.K.)), and the number of steps, active steps, and moderate-intensity activity time were confirmed every two weeks. Regarding the health status of the subjects, after 4, 8, and 12 weeks of intake, a questionnaire and a doctor's examination were conducted to understand the occurrence of subjective and objective symptoms, etc.

[0096] 3) Fatigue and sleep evaluation

[0097] Regarding fatigue, at the start of intake, after 4, 8, and 12 weeks of intake, the subjects were evaluated at home using the VAS (Visual Analogue Scale) method upon waking up. Regarding sleep evaluation, at the start of intake, after 4, 8, and 12 weeks of intake, the subjects were evaluated at home using the OSA (obstructive sleep apnea) sleep questionnaire MA version upon waking up.

[0098] 4) Blood tests and urine tests

[0099] Blood tests were performed before intake, and 4, 8, and 12 weeks after intake to examine white blood cells (WBC), red blood cells (RBC), hemoglobin (Hb), hematocrit (Ht), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelets (Plt), white blood cell classification, total protein, albumin (Alb), albumin / globulin ratio (A / G ratio), aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (γ-GTP), creatinine, blood sugar, total cholesterol (T-Cho), high-density lipoprotein cholesterol (HDL-Cho), low-density lipoprotein cholesterol (LDL-Cho), neutral fat (TG), creatine phosphokinase (CPK), uric acid (UA), urea nitrogen (UN), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), sodium (Na), potassium (K), chlorine (Cl), calcium (Ca), magnesium (Mg), phosphorus (P), total bilirubin (T-Bil), glycated hemoglobin (HbA1c), glycated albumin, insulin, ketone body classification (total ketone body, acetoacetate, 3-hydroxybutyric acid). Urine tests were performed before intake and 12 weeks after intake to examine sugar, protein, occult blood, and urobilinogen.

[0100] 5. Statistical Analysis

[0101] The primary evaluation item was visceral fat area in abdominal CT scan. The secondary evaluation items were body weight, BMI, subcutaneous fat area in abdominal CT scan, total fat area, body fat rate in InBody 770, body fat mass, muscle mass, body fat mass of each part, basal metabolic rate, waist circumference, hip circumference, waist / hip ratio, VAS (fatigue), OSA sleep questionnaire MA version, ketone body classification in blood tests, T-Cho, LDL-Cho, HDL-Cho, TG, blood sugar, HbA1c, insulin, and glycated albumin.

[0102] Statistical analysis was performed using statistical processing software SPSS Ver. 26 (IBM Japan, Inc.) with a significance level of 5% in a two-sided test. In addition, in order to confirm the tendency of the subject attributes, the subjects were divided into three groups according to the BMI before intake: a BMI of less than 23 kg / m2, a BMI of 23 kg / m2 or more and less than 25 kg / m2, and a BMI of 25 kg / m2 or more and less than 30 kg / m2. 2 2 2 2 ​​​Subgroup analyses were performed based on participants and gender. The values ​​at each examination period and the changes from before intake to each examination period were statistically analyzed by group, including the mean and standard deviation. Unpaired t-tests were performed to compare the differences between the means of the tested food group and the control food group at each examination period. Furthermore, Dunnett's multiple comparison tests were performed to compare the differences between the means before intake and at each examination period for each tested food group, comparing the differences during the intake period. Paired t-tests were performed for abdominal CT scans to compare the differences during the intake period. Note that the paired t-tests were not corrected for multiples.

[0103] (result)

[0104] 1. Analyst of the object

[0105] The trial initially included 81 participants (41 in the test food group and 40 in the control food group). However, after randomization, one participant in the test food group discontinued the trial due to starting treatment that violated exclusion criteria (2), so the trial started with 80 participants. After the start of the test food intake, one participant in the test food group was found to have violated exclusion criteria (15) after the start of the trial, and one participant in the control food group did not consume any test food and discontinued the trial for personal reasons. After excluding these two participants, the remaining 78 participants (39 in the test food group and 39 in the control food group) were included in the safety analysis population (FAS). In addition, during the period of test food intake, one participant in the test food group discontinued the trial due to the onset of autoimmune pancreatitis, and one participant in the control food group discontinued the trial for personal reasons. Therefore, the efficacy analysis population (PPS) consisted of 76 participants (38 in the test food group and 38 in the control food group). Furthermore, in the control food group, the reliability of data for some evaluation items was questionable, so only these items were excluded from the analysis, resulting in two cases of partial exclusion. One case had LDH, K, and insulin levels excluded from the analysis due to hemolysis in the blood sample, which occurred 4 weeks after ingestion. Another case had blood glucose and insulin levels excluded from the analysis 8 weeks after ingestion, suspected due to dietary intake on the day of the test (non-compliance with management guidelines). The background of the participants in the validity analysis is shown in Table 2.

[0106] [Table 2]

[0107]

[0108] In any of the FAS, PPS, BMI-based subgroups, and sex-based subgroups, no significant differences were found between the test food group and the control food group in terms of sex at the time of consent, age, visceral fat area at the time of screening, BMI, waist circumference, and hip circumference as allocation factors.

[0109] 2. Analysis Results

[0110] (abdominal fat area)

[0111] Changes in visceral fat area, subcutaneous fat area, and total fat area in the PPS are shown in Table 3. No significant difference was found in the visceral fat area, which was the main evaluation item, between the test food group and the control food group. In the test food group, the visceral fat area and the total fat area were significantly reduced after 12 weeks of intake, compared to before the intake.

[0112] [Table 3]

[0113]

[0114] (body measurement items)

[0115] Changes in body weight, BMI, body fat mass, muscle mass, body fat mass of each part, waist circumference, and hip circumference in the PPS, and blood pressure and pulse rate in the FAS are shown in Table 4. For blood pressure and pulse rate, after 4 weeks of intake, the systolic blood pressure, diastolic blood pressure, and the amount of change in pulse rate were significantly higher in the test food group than in the control food group, but no significant fluctuations were found thereafter. For the other evaluation items, no significant differences were found between the test food group and the control food group.

[0116] [Table 4-1]

[0117]

[0118] [Table 4-2]

[0119]

[0120] (blood tests and urine tests)

[0121] Changes in blood test items (secondary evaluation items) in the PPS are shown in Table 5. For T-Cho, after 4 weeks of intake, a significant decrease in the amount of change from before the intake was observed in the test food group compared to the control food group. For blood glucose, a significantly higher value was observed in the test food group compared to the control food group from before the intake, and a significantly higher value was observed even after 4 weeks of intake, but no differences were found thereafter. For insulin values, a significant decrease was observed in the test food group compared to the control food group. In addition, among the other blood test items and urine test items in the FAS, no changes in clinical problems were found in the subjects.

[0122] [Table 5]

[0123]

[0124] (activity meter)

[0125] For the amount of activity during the test period, the number of steps, the number of effective steps, and the time of moderate-intensity activity were confirmed every two weeks, but no significant difference was found between the test food group and the control food group.

[0126] (Subgroup analysis)

[0127] The BMI before the start of the intake was 23 kg / m 2 or more but less than 25 kg / m 2 The results of the subgroup analysis of the above are shown in Table 6. In the subgroup analysis of the BMI before the start of the intake was 23 kg / m 2 or more but less than 25 kg / m 2 In the evaluation items of the abdominal fat area in the subgroup analysis of the BMI before the start of the intake was 23 kg / m 2 or more but less than 25 kg / m 2 In addition, in the evaluation items of the body measurements, the change in the body weight and the BMI from before the start of the intake was significantly reduced in the test food group compared to the control food group after 12 weeks of the intake. Furthermore, in the evaluation items of the blood tests, the total ketone body, acetoacetic acid, and 3-hydroxybutyric acid were significantly increased in the change from before the start of the intake in the test food group compared to the control food group after 12 weeks of the intake. HDL-Cho was significantly increased after 4, 8, and 12 weeks of the intake. TG was significantly decreased in the change from before the start of the intake, and was significantly reduced in the value after 12 weeks of the intake. The blood glucose value was significantly increased in the values before the intake, after 4 and 8 weeks of the intake. Insulin was significantly increased in the change from before the start of the intake, and was significantly suppressed after 12 weeks of the intake.

[0128] [Table 6-1]

[0129]

[0130] [Table 6-2]

[0131]

[0132] In the subgroup analysis of the BMI before the start of the intake was 25 kg / m 2 or more but less than 30 kg / m 2 In the subgroup analysis by gender, no significant difference was found between the test food group and the control food group in all evaluation items.

[0133] (Investigation)

[0134] In the present test, regarding the body fat reducing effect of the licorice extract-containing food, the BMI before the start of the intake was 23 kg / m 2 or more but less than 30 kg / m 2 , and the abdominal visceral fat area was 100 cm 2The above healthy adult men and women were subjected to a randomized placebo-controlled double-blind parallel group comparison test, and the subjects were administered one capsule containing 100 mg of licorice extract (test food) or a capsule not containing licorice extract (control food) once a day for 12 weeks. As a result, in the subgroup of BMI 23 kg / m 2 or less than 25 kg / m 2 , the test food group showed a significantly lower value in the visceral fat area after 12 weeks of ingestion, and a significantly reduced amount of change in the subcutaneous fat area and total fat area, and the amount of change in body weight and BMI from before the start of the test, as compared with the control food group, thus indicating that licorice extract also has a body fat reducing effect in humans, accompanied by a decrease in body weight and BMI.

[0135] As for the mechanism of the body fat reducing effect of licorice extract, it was confirmed that it promotes the decomposition of TG in fat cells in a cell test. In an animal test, it was also confirmed that it inhibits the accumulation of body fat or reduces the size of fat cells, and results supporting the results of the cell test were reported. In this test, a body fat reducing effect was also found, and a significant increase was found in the test food group, as compared with the control food group, in the total ketone body, acetoacetate, and 3-hydroxybutyrate. The elevation of ketone bodies in the blood is an index of the consumption of fatty acids due to β oxidation, and is also seen when insulin resistance is shown, but no increase in blood glucose value or insulin value was observed, and it is considered that it is not due to the exacerbation of insulin resistance, but is due to the acceleration of energy consumption as reported in other functional foods, etc.

[0136] These results indicate that, by ingesting licorice extract, when TG is decomposed in fat cells, free fatty acids are released and are rapidly consumed as an energy source through β oxidation, thus bringing about a body fat reducing effect.

[0137] It was confirmed that, for the decomposition of TG in fat cells, crude kanzakion B in licorice extract showed a contribution rate of almost 100%. From the above, it is considered that the body fat reducing effect due to the ingestion of licorice extract is caused by crude kanzakion B.

[0138] In this test, the safety of licorice extract was also confirmed. As a result, no adverse events due to the ingestion of licorice extract were found, including abnormalities in the values of clinical tests, and it is considered that there is no problem in the safety of long-term ingestion of licorice extract.

[0139] (Conclusion)

[0140] It was shown that a food containing licorice extract is effective in reducing body fat in people who are slightly obese (BMI 23 kg / m 2 or less than 25 kg / m 2 . The abdominal visceral fat area was 100 cm 2The above) had a body fat-reducing effect in healthy adult men and women. In addition, as part of the mechanism of action, it is believed that there is an increased breakdown of TG in fat cells, with an accompanying increase in fatty acid consumption (beta oxidation).

[0141] <Experiment 2>

[0142] Five-week-old male C57BL / 6J mice were divided into groups of six and fed MF (Oriental Yeast), high-fat high-sugar powdered feed (hereinafter referred to as HFS, D12079BM, Research Diet), or HFS mixed with licorice extract, for eight weeks. After eight weeks of intake, body weight and visceral fat weight (the sum of epididymal fat weight, perirenal fat weight, and mesenteric fat weight) were measured. In addition, as the licorice extract, the roots and rhizomes of licorice were pulverized to obtain a powder, and the liquid obtained by the 50% (v / v) ethanol extraction step was dried and solidified using an evaporator and freeze drying, and then pulverized to obtain a licorice extract. These were mixed at 0.3% in the feed and fed. The amount of crude glycyrrhizin B contained in the licorice extract and the amount of body weight gain and visceral fat weight of the mice are shown in Table 7.

[0143] [Table 7]

[0144]

[0145] (Conclusion)

[0146] The more the mice ingested the formulated amount of crude glycyrrhizin B, the slower the body weight gain, and the lighter the visceral fat weight. This indicates that crude glycyrrhizin B is largely involved in the body fat and visceral fat reducing effects of licorice extract, and is also related to the results of Test Example 1.

[0147] <Experiment 3> Preparation of crude glycyrrhizin B

[0148] A licorice extract aqueous solution was prepared by dissolving 10 ml of distilled water per 1 g of licorice extract. Half the volume of ethyl acetate was added to this aqueous solution and mixed, and the mixture was brought to a state in which it was separated into two layers, and the ethyl acetate layer was recovered from this and the operation was repeated three times. Anhydrous sodium sulfate was added to the obtained ethyl acetate layer, and an organic fraction was obtained by filtration. The organic fraction was concentrated using an evaporator, and the resulting fraction was repeatedly purified by high-performance liquid chromatography using an ODS column (acetonitrile: water = 60:40, 45:55 (both v / v)) using hexane: ethyl acetate = 1:1 (v / v) for elution, thereby obtaining the compound (crude glycyrrhizin B). The crude glycyrrhizin B (GB) obtained here was used as the test substance in the following test using mouse preadipocyte 3T3-L1 cells.

[0149] In addition, the above-mentioned licorice extract can be obtained according to the following method. That is, the stems, roots, and rhizome portions of licorice are pulverized to obtain a licorice powder. 50 g of the licorice powder is extracted with 500 mL of 50% aqueous ethanol at 20°C, and the obtained extract is frozen and dried with an evaporator to obtain a 50% aqueous ethanol extract of licorice.

[0150] The spectral data of the compound are shown below, and are highly consistent with the spectral data of glabridin B reported in the literature (Zeng L. et al., Heterocycles 34:575-587 (1992)) isolated from plants of the genus Glycyrrhiza, and are not contradictory to the structural information, and thus the compound is identified as glabridin B (GB).

[0151]

[0152]

[0153] ESI+-MS m / z: 393.13059 [M+Na]+ (calcd for C21H22O6Na: 393.13086)

[0154] Glabridin B (GB) contained in the licorice extract was quantified using the high-performance liquid chromatograph method described in detail below. As a result, the content of glabridin B (GB) was 132.6 μg per 1 g of licorice extract. In addition, the glycyrrhizic acid contained in the above-mentioned licorice extract was 123 mg per 1 g of licorice extract, and the value (ratio) of the content of glabridin B (GB) to the content of glycyrrhizic acid was 0.00107.

[0155] (Analysis conditions of glabridin B (GB))

[0156] Column: TSKgel ODS-100V 5 μm (Tosoh) 4.6 mm (inner diameter) x 150 mm (length)

[0157] Column temperature: 40°C, mobile phase: acetonitrile: water = 45:55 (v / v)

[0158] Flow rate: 1 ml / min

[0159] Detection wavelength: 290 nm

[0160] Retention time of GB: 33.2 minutes

[0161] <Experiment 4> Study of the fat decomposition effect of licorice extract and glabridin B (GB)

[0162] Mouse preadipocytes (3T3-L1) were cultured for 2 days in differentiation-inducing medium (0.5 mM Isobutyl-methylxanthine, 1 μM Dexamethasone, 1 μg / mL Insulin in 10% FBS / DMEM), then cultured for 7 days in 10% FBS / DMEM medium containing 1 μg / mL Insulin. On day 9 of differentiation induction, the test substances (licorice extract and GB) were added, followed by 19 hours of culture. The licorice extract used in Experiment 1 (final concentrations of 100 μg / mL, 200 μg / mL, and 300 μg / mL) was used as the licorice extract, and the GB prepared in Experiment 1 (final concentrations of 13.5 ng / mL, 27 ng / mL, and 40 ng / mL) was used as the GB. Three cells were included in each group. Glycerol released into the culture medium from the breakdown of accumulated neutral lipids in the cells was quantified using laboratory assays (TM) for triglycerides (Fujifilm and Kodenki). The results are presented below. Figure 1 Glycerin levels are expressed as mean ± standard deviation. Figure 1 The symbols in the table represent the following: ** p<0.01 vs. no treatment (Student's t-test).

[0163] like Figure 1 As shown, the increased amount of glycerol released from adipocytes in a concentration-dependent manner by GB indicates that intracellular lipolysis occurs in accordance with GB concentration, suggesting that GB promotes lipolysis in adipocytes. On the other hand, licorice extract also releases glycerol from adipocytes, but the amount of glycerol released is less at a concentration of 300 μg / mL compared to 100 μg / mL and 200 μg / mL. Furthermore, the GB content of licorice extract at concentrations of 100 μg / mL, 200 μg / mL, and 300 μg / mL is 13.5 ng / mL, 27 ng / mL, and 40 ng / mL, respectively. Licorice extract also exhibits excellent lipolysis effects, but the effect decreases with increasing concentration, indicating that licorice extract contains various components, including those that may inhibit lipolysis. Therefore, to achieve a higher lipolysis effect, a purified form of GB is preferable to licorice extract.

[0164] <Experiment 5> Effects of licorice extract on body weight, BMI, and visceral fat levels (an indicator of visceral fat accumulation).

[0165] One subject (a 59-year-old male) ingested 200 mg of the licorice extract used in Experiment 1 daily for 100 days. Subjects were provided with recording paper to record their weight, BMI, and visceral fat levels as measured using a TANITA body composition analyzer on days 0, 30, 45, 72, 83, 92, and the day after the final intake date (day 101). Changes in weight, BMI, and visceral fat levels before the start of intake are shown below. Figure 2 .

[0166] like Figure 2 As shown, by ingesting licorice extract containing GB, a decrease in visceral fat levels was first observed, followed by a decrease in weight and BMI. Combined with the results of Experiment 3 above, it is concluded that GB in licorice extract efficiently breaks down fat in adipocytes, reducing visceral fat and other body fat.

[0167] <Experiment 6>

[0168] Five-week-old male C57BL / 6J mice were divided into groups of six and fed MF (Oriental yeast), high-fat, high-sugar powdered diet (hereinafter referred to as HFS, D12079BM, Research Diet), or HFS mixed with licorice extract for eight weeks. After eight weeks of feeding, visceral fat weight (the sum of periepididymal fat, perirenal fat, and mesenteric fat weight) was measured. Additionally, as licorice extract (Examples 1-5), licorice root and stolons were pulverized to obtain powder. The liquid obtained from the 50% (v / v) ethanol extraction step was dried and solidified using an evaporator and freeze-dried, then pulverized to obtain licorice extract. These were mixed into the diet at the amounts (%) shown in Table 8 below and fed to the mice. The human equivalent of crude glycyrrhizin dosage (μg / day) and the visceral fat weight inhibition rate (%) in mice are shown together in Table 8.

[0169]

[0170] (in conclusion)

[0171] As shown in the table above, under the condition that the dosage (intake) of crude glycyrrhizin in mice was converted to 9.7 μg / day to 47.6 μg / day for human use, a significant inhibitory effect on visceral fat weight was observed.

[0172] Industrial availability

[0173] The internal composition of the present application has an excellent fat decomposition promoting effect by containing picroliv B, and has a significant weight loss effect. By using the internal composition of the present application, fat in fat cells can be efficiently decomposed, and body fat such as subcutaneous fat, visceral fat, etc. can be reduced, and finally a significant weight loss effect, a BMI improvement effect can be obtained. In addition, an effect of improving insulin resistance, lowering cholesterol value can also be obtained. Therefore, according to the internal composition of the present application, obesity or lifestyle diseases can be prevented and improved for a long time, and maintenance and improvement of health can be achieved. Furthermore, since the content of picroliv B in the plant extract becomes an index of the effectiveness of the plant extract on fat decomposition, weight loss, etc., a plant extract with high effectiveness can be efficiently selected.

Claims

1. An internal compound containing crude glycyrrhizin B, used for any one or more of the following: fat decomposition, fat burning, fat consumption assistance, BMI improvement, BMI reduction assistance, abdominal fat reduction, visceral fat reduction, body fat reduction, waist circumference reduction, improvement of insulin resistance, and reduction of LDL cholesterol levels.

2. An internal compound containing an effective amount of crude glycyrrhizin B, for use in any one or more of the following: fat decomposition, fat burning, fat consumption assistance, BMI improvement, BMI reduction assistance, abdominal fat reduction, visceral fat reduction, body fat reduction, waist circumference reduction, improvement of insulin resistance, and reduction of LDL cholesterol levels.

3. The oral composition according to claim 2, wherein the effective amount is more than 10 μg per day for adults.

4. The internal component according to claim 1 or 2, used for individuals with a BMI of 23 kg / m². 2 Above but below 30kg / m 2 Adults.

5. The internal component according to claim 1 or 2, wherein the area for visceral fat is 100 cm². 2 Adults above that age.

6. The oral composition according to claim 1 or 2, wherein the dosage is 5 to 100 μg / day, based on the total amount of crude glycyrrhizin B.

7. A method for determining effectiveness, which uses the content of crude glycyrrhizin B as an indicator to determine the effectiveness of plant extracts in fat breakdown and / or weight reduction in organisms.

8. The method for determining the effectiveness of fat breakdown and / or weight reduction according to claim 7, wherein the plant is licorice.

9. An indicator, which is a quality indicator for plant extracts, characterized in that, The aforementioned quality is related to the effectiveness of fat breakdown and / or weight reduction in organisms, and is composed of glycyrrhizin B.

10. The indicator according to claim 9, wherein the plant is licorice.

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