WO2025037604A1 - Internal composition for lipolysis and/or weight loss, method for determining effectiveness, and index for quality of plant extract - Google Patents

Abstract

The present invention provides a novel preparation which has an excellent lipolysis-promoting effect, an excellent visceral fat and body fat loss effect, an excellent weight loss effect, and an excellent BMI improvement effect. Another purpose of the present invention is to provide a novel preparation which has an excellent insulin resistance amelioration effect, an excellent cholesterol level reduction effect, and the like.　The present invention is an internal composition which contains glyasperin B and which is used for at least one of lipolysis, fat burning, fat consumption support, BMI improvement, BMI reduction support, abdominal fat loss, visceral fat loss, body fat loss, waist circumference reduction, insulin resistance amelioration, and LDL cholesterol level reduction. The internal composition according to the present invention is preferably used by adults who have a BMI of at least 23 kg/m² but less than 30 kg/m² and adults who have a visceral fat area of at least 100 cm².

Description

Compositions for internal use on lipolysis and/or weight loss, methods for determining efficacy, and indicators of quality of plant extracts

The present invention relates to an internal composition for lipolysis and/or weight loss, a method for determining efficacy, and an indicator of the quality of a plant extract.

Body fat occurs when energy intake is in excess of energy expenditure, and the excess accumulates as neutral fat in white fat cells. Excessive fat accumulation leads to obesity and weight gain, which not only causes various lifestyle-related diseases but also poses a major cosmetic problem. Obesity with a large accumulation of visceral fat has been linked to conditions such as insulin resistance and arteriosclerosis, while obesity with a large accumulation of subcutaneous fat is a major concern for both men and women from the perspective of cosmetic care.

Traditionally, various measures to prevent and improve obesity have been taken, such as restricting energy intake, dietary restrictions, and the search for substances that inhibit carbohydrate absorption in the digestive tract. However, restricting energy intake can also cause a decrease in basal metabolic rate, and obesity may not improve. Restricting energy intake can also lead to a deficiency of necessary nutrients, which can be harmful to health. For this reason, it has come to be thought that the ideal way to eliminate obesity is to actively break down accumulated fat and release it as heat energy, thereby allowing one to maintain an ideal body weight. Under these circumstances, in recent years, there has been active research into functional ingredients that promote lipolysis among food ingredients, and many foods and beverages have been proposed.

For example, it is known that a mixture prepared by combining salmon milt extract, brewer's yeast extract, young barley leaf extract, and chicken collagen (chicken collagen) has an excellent lipolysis promoting effect (Patent Document 1), that at least one of an extract of Betulaceae (Betulaceae) and an extract of Kumazasa (Poaceae) improves obesity and prevents the increase of adipose tissue by promoting the decomposition of fat in whole body or local adipose tissue (Patent Document 2), and that certain plants such as Juniperus communis or their extracts are useful as medicines, foods, or cosmetics that promote the decomposition of neutral fats accumulated in adipose tissue and exert slimming effects such as suppressing, preventing, or improving obesity (Patent Document 3). However, the lipolysis and weight loss effects of extracts of these plants are not sufficient at present. Patent Document 4 also discloses that a composition combining a licorice hydrophobic extract with an antioxidant component and the like has the effects of suppressing body fat accumulation, promoting body fat decomposition, and promoting energy production, but a sufficient lipolysis promoting effect is not obtained with licorice hydrophobic extract alone.

JP 2011-074051 A JP 2006-045120 A JP 2012-229266 A International Publication No. 2008/143182

The problem that the present invention aims to solve is to provide a new formulation that is excellent in promoting lipolysis, reducing visceral fat and body fat, reducing body weight, and improving BMI. Another object of the present invention is to provide a new formulation that is excellent in improving insulin resistance, reducing cholesterol levels, etc.

As a result of intensive research, the inventors have found that "gliasperin B," a specific component in licorice extract, has an excellent effect of promoting lipolysis. They have also found that a preparation containing licorice extract containing gliasperin B has an excellent effect of promoting lipolysis, as well as an effect of reducing visceral fat and body fat, a weight loss effect, an effect of improving BMI, an effect of improving insulin resistance, and an effect of reducing cholesterol levels. They have also found that the content of gliasperin B in a plant extract is an indicator of the effectiveness of the plant extract for lipolysis, weight loss, etc. The gist of the present invention is as follows.

[1] An internal composition containing gliasperin B which is used for one or more of the following: decomposing fat, burning fat, assisting in fat consumption, improving BMI, assisting in lowering BMI, reducing abdominal fat, reducing visceral fat, reducing body fat, reducing waist circumference, improving insulin resistance, and reducing LDL cholesterol levels.
[2] An internal composition containing an effective amount of gliasperin B and used for one or more of the following: decomposing fat, burning fat, assisting in fat consumption, improving BMI, assisting in lowering BMI, reducing abdominal fat, reducing visceral fat, reducing body fat, reducing waist circumference, improving insulin resistance, and reducing LDL cholesterol levels.
[3] The oral composition described in [2], wherein the effective amount is 10 μg or more per day for an adult.
[4] The internal composition described in [1] or [2], which is used for adults with a BMI of 23 kg/ ^m2 or more and less than 30 kg/ ^m2 .
[5] The internal composition described in [1] or [2], which is used for adults with a visceral fat area of 100 ^cm2 or more.
[6] The internal composition according to [1] or [2], which is taken at a dose of 5 to 100 μg/day in terms of the total amount of gliaperin B.
[7] A method for determining the effectiveness of a plant extract for lipolysis and/or weight loss in a living body, using the content of gliasperin B as an indicator.
[8] The method for determining the effectiveness of a compound according to [7] above, wherein the plant is licorice.
[9] An indicator of the quality of a plant extract, the quality being related to effectiveness in lipolysis and/or weight loss in a living body, characterized in that the indicator consists of gliasperin B.
[10] The indicator described in [9], wherein the plant is licorice.

The internal composition of the present invention contains gliasperin B, preferably as an active ingredient, functional ingredient, indicator ingredient or contributing ingredient, and thus has an excellent lipolysis promoting effect and a remarkable weight loss effect. By using the internal composition of the present invention, fat in adipocytes can be efficiently broken down and body fat such as subcutaneous fat and visceral fat can be reduced, resulting in a remarkable weight loss effect and an improvement effect on BMI. In addition, the internal composition of the present invention can also improve insulin resistance and reduce sterol levels. Therefore, the internal composition of the present invention can prevent and improve obesity and lifestyle-related diseases in the long term, and can achieve the maintenance and improvement of health. Furthermore, since the content of gliasperin B in a plant extract is an indicator of the effectiveness of the plant extract for lipolysis, weight loss, etc., it becomes possible to efficiently select a highly effective plant extract.

FIG. 1 shows the lipolysis promoting effect of licorice extract and gliasperin B. FIG. 1 shows the effect of licorice extract on body weight, BMI and visceral fat level.

Below, the internal composition of the present invention, the method for determining the effectiveness of the composition on lipolysis and/or weight loss, and the quality indicators of the plant extract are described in detail.

<Composition for internal use>
The internal composition of the present invention contains gliasperin B. Although not limited thereto, it is preferable that gliasperin B is contained as an active ingredient, a functional ingredient, an indicator ingredient or a contributing ingredient. In addition to gliasperin B, which is an essential ingredient, the internal composition of the present invention may contain other ingredients within a range that does not impair the effects of the present invention. The internal composition of the present invention contains gliasperin B and other ingredients, and the form of the internal composition of the present invention, etc. will be described below.

In this specification, an active ingredient refers to a pharmacologically active ingredient or a physiologically active ingredient contained in a drug, quasi-drug, etc. that exerts a desired effect.

In this specification, functional ingredients refer to ingredients contained in internal compositions such as foods that contribute to a specific health purpose. Specifically, the mechanism of action related to functionality has been considered through in vitro tests, in vivo tests, or clinical trials (human tests), and the ingredients can be directly or indirectly qualitatively and quantitatively confirmed. Functional foods are provided to consumers with a display of functions based on the effects of the ingredients contained in the functional food, and when providing such foods to consumers as functional foods, the content of the functional ingredients and the method of quantifying them must be notified.

In this specification, the term "index ingredient" refers to an extract or essence that is considered to be a functional ingredient when a specific ingredient that can explain part of the scientific basis of functionality is known, but the functionality cannot be fully explained by the specific ingredient alone, and the mechanism of action related to the functionality to be displayed has been considered for at least one ingredient through in vitro tests, in vivo tests, or clinical tests (human tests).

In this specification, a contributing ingredient refers to a substance that, when incorporated, directly or indirectly affects the physiological functions of the body. In foods for specified health uses and foods with functional claims, the equivalent of a contributing ingredient could be, but is not limited to, a "contributing ingredient," a "functional contributing ingredient," or an "indicator ingredient." In addition, the equivalent of a contributing ingredient in medicines and quasi-drugs is an "active ingredient." Furthermore, in the sale of food compositions other than foods with functional claims, this concept also includes ingredients that are implicitly linked to an effect and emphasized to consumers in the form of "Contains XX ingredient. For burning calories."

(Gliasperin B)
Glyasperin B (hereinafter also referred to as "GB") is an isoflavanone derivative represented by the following formula: It is 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.

Gliasperin B is a component that has been confirmed to be contained in licorice extract, and the gliasperin B in the present invention is preferably derived from licorice extract. It has been known that licorice extract has a lipolysis promoting effect, and there have been documents suggesting that glabridin, which is contained in large amounts in licorice extract, may have a lipolysis promoting effect (see Patent Document 4, etc.). However, it was not clear which component in licorice extract has the lipolysis promoting effect. The present inventors have discovered for the first time that, of the many components present in licorice extract, gliasperin B has a lipolysis promoting effect. They have then completed the invention of an internal composition that contains this gliasperin B and has the effects of decomposing fat, burning fat, assisting in fat consumption, improving BMI, assisting in lowering BMI, reducing abdominal fat, reducing visceral fat, reducing body fat, reducing waist circumference, improving insulin resistance, and reducing LDL cholesterol levels.

The gliasperin B in the present invention may be derived from licorice or may be a chemically synthesized product. In this specification, when we say that it contains gliasperin B, this also includes the case where it contains a plant such as licorice that contains gliasperin B.

The type of licorice is not particularly limited, but may be any plant of the genus Glycyrrhiza in the family Fabaceae, such as Glycyrrhiza uralensis (G. uralensis Fisch. et DC; ural licorice), Glycyrrhiza inflata (G. inflata BAT.; butterfly licorice), Glycyrrhiza glabra (G. glabra L.; licorice), Glycyrrhiza glabra L. ( ... These include G. glabra L. var glandu lifera Regel et Herder (Chinese tallow tree), G. aspera, G. echinata L. (Chinese tallow tree), and G. pallidiflora Maxim (Chinese tallow tree).

Methods for obtaining gliasperin B derived from licorice include 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 crushed part of the plant (e.g., roots, stems, stolons, leaves, flowers, fruits, etc.) with a solvent, and the obtained 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. Of these, the extraction solvent is preferably water, alcohol, or aqueous alcohol, and more preferably hot water, ethanol, or aqueous 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, even more 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), even more preferably 40% (v/v) to 60% (v/v), and particularly preferably 50% (v/v).

Distilled water is added to the dried licorice extract to dissolve it, and an aqueous licorice extract solution is prepared. An organic solvent such as ethyl acetate is added in an amount equal to half the volume of the aqueous solution, and the solution is mixed to separate into two layers, from which the organic solvent layer such as ethyl acetate is collected. This operation is repeated several times, preferably about three times. Anhydrous sodium sulfate is added to the obtained organic solvent layer, and the organic fraction can be obtained by filtering. The organic fraction is concentrated using an evaporator, and the fraction eluted with hexane:ethyl acetate = 1:1 (v/v) or the like using a silica gel column chromatograph is repeatedly purified using high-performance liquid chromatography (acetonitrile:water = 60:40, 45:55 (both v/v)) using an ODS column to obtain the compound (gliasperin B).

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

(Other ingredients)
The internal composition of the present invention may contain, in addition to gliasperin B, a carrier, an excipient, a solvent, and other optional components within a range that does not impair the effects of the present invention.

When the internal composition of the present invention contains gliasperin B, it may contain gliasperin B purified from licorice extract as described above or a chemically synthesized gliasperin B, or it may contain licorice extract so that the final content of gliasperin B in the internal composition is within the above numerical range. From the viewpoint of promoting lipolysis, the internal composition of the present invention preferably contains gliasperin B within the above numerical range and does not contain any licorice-derived components other than gliasperin B, or, if it contains any licorice-derived components other than gliasperin B, the ratio of the gliasperin B content to the glycyrrhizic acid content (the value obtained by dividing the gliasperin B content by the glycyrrhizic acid content) is 0.001 or more, using glycyrrhizic acid, which is a quantitative indicator of licorice extract in the Japanese Pharmacopoeia, as a comparison standard.

The internal composition of the present invention can be used as a food or drink, a functional food, a food for specified health uses, a nutrient-functional food, a cosmetic, a quasi-drug, a medicine, or a cosmetic agent, etc., to efficiently break down fat and reduce body fat, such as subcutaneous fat and visceral fat.

When preparing the internal composition of the present invention as a food or beverage, in addition to gliasperin B, sweeteners, colorants, preservatives, thickeners, stabilizers, gelling agents, pasting agents, antioxidants, colorants, bleaching agents, mold inhibitors (mold inhibitors), yeast food, gum base, flavorings, acidulants, seasonings, emulsifiers, pH adjusters, kansui, leavening agents, nutritional enhancers, and other food and beverage ingredients can be mixed and prepared into the desired form. When the internal composition of the present invention is made into a food or beverage form, there are no particular limitations on the form. Examples include supplement-type foods such as gels, granules, fine granules, capsules, tablets, powders, liquids, and semisolids; beverages such as carbonated drinks, soft drinks, milk drinks, alcoholic drinks, fruit juice drinks, teas, and nutritional drinks; powdered drinks such as powdered juice and powdered soup; confectioneries such as gum, tablets, candies, cookies, gummies, rice crackers, biscuits, and jellies; bread, noodles, cereals, jams, and seasonings. These foods are used as foods and beverages to efficiently break down fat and reduce body fat such as subcutaneous fat and visceral fat, and can be used, for example, as nutraceuticals such as dietary supplements, functional foods, foods for specified health uses, and foods for the sick, in addition to general foods and beverages.

When the internal composition of the present invention is prepared as a medicine (including quasi-drugs), in addition to gliasperin B, other medicinal ingredients, pharma- ceutical acceptable carriers, additives, etc. may be optionally blended, if necessary. Specific examples of pharma- ceutical acceptable carriers and additives include binders, disintegrants, lubricants, wetting agents, buffers, preservatives, and flavorings. When the internal composition of the present invention is prepared as a medicine, its form is not particularly limited. Examples include injections, external preparations, inhalants, suppositories, films, lozenges, liquids, powders, tablets, granules, capsules, syrups, eye drops, eyewashes, and nasal drops. Among these, forms suitable for oral administration (i.e., internal medicines) are preferred, and specific examples of such forms include lozenges, liquids, powders, tablets, granules, capsules, and syrups. These medicines (including quasi-drugs) are used as medicines to efficiently break down fat and reduce body fat, including subcutaneous fat and visceral fat.

Gliasperin B contained in the internal composition of the present invention can also be used as a cosmetic (including functional cosmetic) or topical quasi-drug. To prepare a cosmetic or topical quasi-drug, a pharma- ceutical or cosmetically acceptable carrier (water, oily components, etc.) is added to Gliasperin B and the cosmetic is prepared in the desired form. The cosmetic is not particularly limited in form 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, spray, liniment, pack, etc. These cosmetic products are used as cosmetic products for efficiently breaking down fat and reducing subcutaneous fat, etc.

The dosage and application amount of the internal composition of the present invention can be appropriately determined depending on the age, body weight, health condition, degree of obesity, disease state, etc. of the user, but is not limited to, and can be, in terms of the total amount of gliaperin B, 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 per day for an adult, and can be 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, etc. For example, the total amount of gliasperin B taken per day by an adult is 0.1 μg to 5000 μg, preferably 0.5 μg to 1000 μg, more preferably 1 μg to 500 μg, even more preferably 1 μg to 200 μg, even more preferably 1 μg to 100 μg, particularly preferably 5 μg to 100 μg, even more particularly preferably 5 μg to 60 μg, even more particularly preferably 10 μg to 50 μg, and most preferably 10 to 20 μg.

When the internal composition provided by the present invention is orally administered to humans, it is preferable to administer an effective amount of gliasperin B. In this specification, an effective amount refers to the dosage at which the desired effect is observed, whether it is a direct or indirect action. The effective amount varies depending on age and body weight, but can be, for example, 1 μg or more, 3 μg or more, 5 μg or more, 7 μg or more, 10 μg or more, or 12 μg or more per day for an adult, calculated as the total amount of gliasperin B described above, or 1 mg or less, 500 μg or less, 200 μg or less, 100 μg or less, 60 μg or less, 50 μg or less, or 20 μg or less. For example, the total amount of gliasperin B per day for an adult is 1 μg to 1 mg, preferably 1 μg to 500 μg, more preferably 1 μg to 200 μg, even more preferably 1 μg to 100 μg, even more preferably 5 μg to 100 μg, particularly preferably 5 μg to 60 μg, even more particularly preferably 10 μg to 50 μg, even more particularly preferably 10 μg to 20 μg, and most preferably 14 μg.

From the viewpoint of easily obtaining a weight loss effect, the internal composition of the present invention is preferably used for adults with a BMI of ²³ kg/m2 or more and less than 30 kg/ ^m2 , and more preferably for adults with a BMI of 23 kg/ ^m2 or more and less than 25 kg/ ^m2 . According to the criteria of the Japan Society for the Study of Obesity, a BMI of 18.5 kg/ ^m2 or more and less than 25 kg/ ^m2 is considered to be normal body weight, and a BMI of 25 kg/ ^m2 or more and less than 30 kg/ ^m2 is considered to be obesity level 1. The internal composition of the present invention is preferably used for adults with a so-called normal body weight to slightly obese (obesity level 1), and is particularly preferably used for adults who are not classified as obese but are slightly obese from a normal body weight. BMI (Body Mass Index) is also called body mass index, and is a body mass index that indicates the degree of obesity, calculated from body weight and height using the following formula. BMI is used as an international index for adults.

Calculation formula: BMI = weight kg ÷ (height m) ²

From the viewpoint of easily obtaining the effects of promoting lipolysis and reducing body weight, the internal use composition of the present invention is preferably used for adults with a visceral fat area of 100 ^cm2 or more. The visceral fat area can be calculated in the usual manner by performing abdominal CT scan and taking multiple slice images at intervals of 1 cm or more centered on the navel.

Another embodiment of the present invention is a weight loss agent, an agent for reducing visceral fat and body fat, an agent for improving BMI, an agent for improving insulin resistance, or an agent for reducing cholesterol levels, which contains gliasperin B as an active ingredient.

The present invention also includes the above-mentioned weight loss agent, visceral fat/body fat reduction agent, BMI improvement agent, insulin resistance improvement agent, or cholesterol level reduction agent, which contain gliasperin B, and thus have excellent fat decomposition promoting effects and a remarkable weight loss effect. By using the weight loss agent, visceral fat/body fat reduction agent, BMI improvement agent, insulin resistance improvement agent, or cholesterol level reduction agent 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, resulting in a remarkable weight loss effect and BMI improvement effect. Insulin resistance improvement and sterol level reduction effects can also be obtained. Therefore, the weight loss agent, visceral fat/body fat reduction agent, BMI improvement agent, insulin resistance improvement agent, or cholesterol level reduction agent of the present invention can prevent and improve obesity and lifestyle-related diseases in the long term, and can achieve the maintenance and promotion of health. In addition, since the weight loss agent, visceral fat/body fat reducing agent, BMI improving agent, insulin resistance improving agent, or cholesterol level reducing agent of the present invention has the same configuration as the above-mentioned internal composition of the present invention containing gliasperin B as an active ingredient, the specific explanation can be applied as is to the explanation in the section on the internal composition of the present invention.

Methods for determining the efficacy of plant extracts for lipolysis and/or weight loss
The present invention also includes a method for determining the effectiveness of a plant extract in lipolysis and/or weight loss in a living body, using the content of gliasperin B as an index. As described above, a preparation containing gliasperin B as an active ingredient has an excellent lipolysis promoting effect and a remarkable weight loss effect. In addition, it is possible to efficiently decompose fat in adipocytes and reduce body fat such as subcutaneous fat and visceral fat, resulting in a remarkable weight loss effect and BMI improvement effect. In addition, it is possible to improve insulin resistance and reduce sterol levels. Since the above effects increase depending on the content of gliasperin B, it is possible to determine the effectiveness of the plant extract in lipolysis and/or weight loss in a living body by using the content of gliasperin B in the plant extract as an index. The above plant extract is not particularly limited as long as it contains gliasperin B, but licorice extract is preferable.

The determination method of the present invention is described in detail below. The determination method of the present invention includes a step of measuring the content of gliasperin B in the plant extract (hereinafter also referred to as the "measurement step"). It is also preferable that the method includes a step of comparing the obtained amount of gliasperin B with a standard amount of gliasperin B (hereinafter also referred to as the "comparison step").

In the above measurement step, the method for measuring the content of gliasperin B in the plant extract is not particularly limited, and can be measured by a method known to those skilled in the art. Examples include Western blotting and ELISA methods. In both the above Western blotting and ELISA methods, an anti-gliasperin B antibody that specifically binds to gliasperin B is used, and the content of gliasperin B can be measured with high accuracy. In the Western blotting method, the intensity of the resulting gliasperin B-specific blot can be compared with the intensity of a standard specimen (positive control) of known concentration, thereby calculating the gliasperin B content in the sample.

Gliasperin B used as a standard in the determination method of the present invention can be obtained, for example, by extraction from a natural source, by expression of a recombinant nucleic acid encoding gliasperin B, or by chemical synthesis.

In the assessment method of the present invention, the comparison step involves comparing the content of gliasperin B in the extract to be evaluated with the content of the standard gliasperin B. The amount of gliasperin B to be used as the standard can be appropriately selected depending on the purpose and subject of the evaluation.

<Indicators of plant extract quality>
The present invention also includes an invention relating to an index of the quality of a plant extract, the quality being related to the effectiveness of the plant extract in lipolysis and/or weight loss, and characterized in that the index is made of gliasperin B. As described above, a preparation containing gliasperin B as an active ingredient has an excellent lipolysis promoting effect and a remarkable weight loss effect. In addition, the preparation can efficiently decompose fat in adipocytes and reduce body fat such as subcutaneous fat and visceral fat, resulting in a remarkable weight loss effect and BMI improvement effect. In addition, the preparation can improve insulin resistance and reduce sterol levels. Since the above effects increase depending on the content of gliasperin B, it is possible to determine the effectiveness of the plant extract in lipolysis and/or weight loss in a living body by using gliasperin B in the plant extract as an index. Licorice is preferable as the plant in the plant extract.

The indicator of the quality of the plant extract of the present invention corresponds to gliasperin B in the above-mentioned method for determining the effectiveness of the plant extract for lipolysis and/or weight loss. The specific explanation of the indicator of the present invention can be applied to the explanation of the above-mentioned determination method.

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

As shown below, a randomized, double-blind, parallel-group comparative study was conducted in subjects to evaluate the effect on visceral fat of taking the test food (a hard capsule formulation containing licorice extract) for 12 weeks, using a control food (hard capsule without licorice extract) as a control.

<Test 1>
1. Test food (composition, packaging, etc.)
The test food contained 100 mg of licorice extract (manufactured by MG Pharma Co., Ltd.; derived from Ural licorice) per capsule, and was composed of other excipients. The control food did not contain licorice extract, and was composed of excipients colored with caramel coloring. All test foods were in the form of white hard capsules, packaged in aluminum pouches of 30 capsules each, and an identification code was displayed on the test foods. The indistinguishability of the test foods and their packaging was reviewed and approved by the IRB. The nutritional components and related components of the test foods are shown in Table 1 below. The amount of gliasperin B contained in the test foods was 14 μg per recommended daily intake.

2. Subjects The target number of subjects in this study was calculated to be 34 per group, for a total of 68 subjects, assuming that the effect size of the test food is large (0.8) in a parallel group comparison study between two groups, the significance level is 0.05, and the detection power is 80-90%, so that 26-35 subjects per group are required. In addition, the number of entries was set at 80 (40 per group) in consideration of cases that may drop out or be discontinued during the study. Subjects were recruited from volunteers registered with the study contracting institution, and 81 subjects (50 men, 31 women) who met the following inclusion criteria, did not violate the exclusion criteria, and were deemed appropriate for participation in this study by the study principal physician were included as subjects.

(Selection Criteria)
For the screening test, participants were required to meet the following conditions:
(1) Men and women aged 20 years or older and younger than 65 years, (2) individuals with a BMI of 23.0 kg/ ^m2 or more and less than 30.0 kg/ ^m2 , (3) individuals with a visceral fat area of 100 ^cm2 or more, (4) individuals who have been fully informed of the purpose and contents of this study, are capable of consent, have a full understanding of the study, voluntarily volunteer to participate, and can agree to participate in this study in writing.
(Exclusion Criteria)
(1) Persons suffering from or with a history of serious cardiovascular, hepatic, renal, respiratory, endocrine or metabolic disorders; (2) Persons receiving treatment for chronic diseases such as dyslipidemia, hypertension or diabetes; (3) Persons with mental illnesses such as depression, schizophrenia or bulimia nervosa; (4) Persons who regularly use medicines or quasi-drugs that may affect the evaluation of this study, such as body fat, triglycerides, body weight, cholesterol and energy metabolism; (5) Persons who regularly use supplements or health foods (specified health foods, functional foods, etc.) that may affect the evaluation of this study, such as body fat, triglycerides, body weight, cholesterol and energy metabolism; (6) Persons who may be allergic to the test foods. (7) Persons with metal in the area where the abdominal CT scan will be performed; (8) Persons with implanted medical devices such as a cardiac pacemaker or an implantable cardioverter defibrillator; (9) Persons with claustrophobia; (10) Persons with a habit of smoking; (11) Persons with a habit of drinking large amounts of alcohol (60 g or more of pure alcohol per day); (12) Persons with extremely irregular eating habits; (13) Persons working night shifts or day-night shifts; (14) Persons who have not had a bowel movement for five days or more; (15) Persons with menopausal symptoms; (16) Persons with hypertension, pseudoaldosteronism, hypokalemia, or myopathy; (17) Persons who regularly use medicines containing licorice or foods that mainly contain licorice; (18) Persons who are prone to edema or weakness; (19) Persons with urinary problems; (20) Persons who have had 200 (21) Those who have participated in other clinical trials within the past four months, those currently participating in other clinical trials, and those who plan to participate in other clinical trials during the study period; (22) Those who are pregnant or breastfeeding, or those who plan to become pregnant; (23) Those who are otherwise deemed inappropriate by the principal investigator of the study.

3. Study protocol This study was a randomized, placebo-controlled, double-blind, parallel-group comparative study. Tests were conducted at the start of intake of the test food, and after 4 weeks, 8 weeks, and 12 weeks of intake. A random number was generated for randomization by the allocation manager, who was not involved in the implementation of the study, and the subjects were assigned to the subjects who were enrolled by the study director. At this time, it was confirmed that there were no significant differences between the groups in the allocation factors of sex and age at the time of consent acquisition, and visceral fat area, BMI, hip circumference, and waist circumference at the time of screening examination.

During the study period, subjects were instructed to take one capsule of the test food (test food for the test food group, control food for the control food group) once a day with water or lukewarm water before dinner. However, if subjects were unable to take the test food before dinner, they were instructed to take one capsule once a day at a time that was possible.

During the study period, subjects were instructed to maintain normal eating and drinking habits as much as possible, and were prohibited from consuming health foods (foods for specified health uses, foods with functional claims, etc.) that may affect the study's evaluations of body fat, neutral fat, body weight, cholesterol, energy metabolism, etc., as well as grapefruit juice. In addition, subjects were instructed to maintain normal alcohol intake habits, and were prohibited from drinking amounts that deviated from normal intake. In addition, subjects were prohibited from consuming alcohol the day before the study, and were prohibited from consuming anything other than water after 9 p.m. In addition, subjects were prohibited from consuming anything other than water four hours before the abdominal CT scan. During the study period, subjects were instructed to maintain normal exercise habits (frequency, type of exercise, etc.), and were prohibited from changing their exercise habits. During the study period, except in emergency cases, permission from the study physician was required before using any new medications. Vaccinations were prohibited for two weeks prior to the study. Blood donation was prohibited during the study period.

This study was conducted in accordance with the spirit of the Declaration of Helsinki (2013) - Tokyo, Venice, Hong Kong, Somerset West, Edinburgh Amendment, Washington, Tokyo Annotation Added, Seoul Amendment, and Fortaleza Amendment - and in accordance with the "Ethical Guidelines for Life Science and Medical Research Involving Human Subjects (established March 23, 2021, enforced June 30, 2021, partially revised March 10, 2022)". Approval was obtained from the Institutional Review Board (IRB) of the Fukuda Naika Clinic, a medical corporation (approval number: IRB-20220716-2). In addition, the clinical trial was registered in the University Hospital Medical Information Network Clinical Trial Registration System (UMIN) (UMIN Study ID: UMIN000048430).

4. Examination items 1) Physical examination and abdominal CT scan Physical examination items included height (only at the time of screening test), body composition evaluation using InBody770 (InBody Japan, Inc.) (weight, BMI, body fat percentage, muscle mass, body fat mass by region, basal metabolism, other body composition component analysis, etc.), waist circumference, hip circumference, blood pressure, and pulse rate, and CT scans were performed on those who met the selection criteria. For weight, DST-210N (Muratec KDS, Ltd.) was used only at the time of CT scan. BMI was calculated from height and weight, and the BMI at the time of CT scan was used as the value before starting intake.

Abdominal CT scans were performed using a Discovery 710 or Discovery MIDR (GE Healthcare Japan, Ltd.), with three slices taken at 1 cm intervals centered on the navel, and the average values of visceral fat area, subcutaneous fat area, and total fat area were calculated. If the kidneys or ilium were included in the image taken at the navel position, a second image was taken centered on the fourth lumbar vertebra.

2) Food and activity records During the test period, subjects were asked to record alcohol intake, drug use, dietary content, etc. in a daily diary. During the test food intake period, subjects were asked to record the intake of the test food. In addition, subjects were asked to record the amount of food consumed in addition to the content of the food consumed for three days prior to the test. During the test period, subjects were asked to wear an activity monitor (SAT-1, Medithink Co., Ltd.) except when sleeping, and the number of steps, active steps, and duration of moderate intensity activity were checked every two weeks. Regarding the health condition of the subjects, interviews and medical examinations were conducted by a doctor 4, 8, and 12 weeks after intake to ascertain the occurrence of subjective and objective symptoms, etc.

3) Fatigue and sleep evaluation Fatigue was evaluated by VAS at the start of intake, and 4, 8, and 12 weeks after intake, when the subjects woke up at home. Sleep was evaluated by the OSA sleep questionnaire MA version at the start of intake, and 4, 8, and 12 weeks after intake, when the subjects woke up at home.

4) Blood and urine tests Blood was taken before intake and 4, 8, and 12 weeks after intake for WBC, RBC, Hb, Ht, MCV, MCH, MCHC, Plt, white blood cell fraction, total protein, Alb, A/G ratio, AST, ALT, γ-GTP, creatinine, blood glucose, total cholesterol (T-Cho), HDL cholesterol (HDL-Cho), LDL cholesterol (LDL-Cho), triglyceride (TG), CPK, uric acid (UA), urea nitrogen (UN), ALP, LDH, Na, K, Cl, Ca, Mg, P, total bilirubin (T-Bil), HbA1c, glycoalbumin, insulin, and ketone body fraction (total ketone bodies, acetoacetic acid, 3-hydroxybutyric acid). Urine tests were performed for sugar, protein, occult blood, and urobilinogen before and 12 weeks after intake.

5. Statistical analysis The primary outcome was visceral fat area in abdominal CT scans. Secondary outcome measures were body weight, BMI, subcutaneous fat area in abdominal CT scans, total fat area, body fat percentage, body fat mass, muscle mass, body fat mass by region, basal metabolic rate, waist circumference, hip circumference, waist/hip ratio, VAS (feeling of fatigue), OSA Sleep Questionnaire MA version, ketone body fraction in blood tests, T-Cho, LDL-Cho, HDL-Cho, TG, blood glucose, HbA1c, insulin, and glycoalbumin.

Statistical analysis was performed using statistical processing software SPSS Ver. 26 (IBM Japan, Ltd.), and the significance level was set at 5% in a two-sided test. In addition, in order to confirm the tendency according to the subject attributes, subgroup analysis was performed according to the subjects with a BMI of 23 kg/ ^m2 or more and less than 25 kg/ ^m2 before the start of intake, the subjects with a BMI of 25 kg/ ^m2 or more and less than 30 kg/ ^m2 , and the gender. The average value and standard deviation of the values at each test time and the change amount from before the start of intake to each test time were tabulated by group, and an unpaired t-test was performed on the difference in the average value between the test food group and the control food group at each test time to compare the differences between the groups. In addition, Dunnett's multiple comparison test was performed on the difference in the average value between before the start of intake and each test time in each test food group to compare the differences due to the intake period of the test food. For abdominal CT, a paired t-test was performed to compare the differences due to the intake period of the test food. Note that no correction for multiplicity was performed for the paired t-test.

(result)
1. Subjects analyzed Eighty-one subjects were enrolled in this study (41 in the test food group, 40 in the control food group), but after randomization, one subject in the test food group discontinued the study due to initiating treatment that violated the exclusion criterion (2), and the study began with 80 subjects. One subject in the test food group who was found to violate the exclusion criterion (15) after the start of the study after starting to take the test food, and one subject in the control food group who had never taken the test food and discontinued the study for personal reasons, were excluded, leaving 78 subjects (39 in the test food group, 39 in the control food group) as the safety analysis set (FAS). In addition, one patient in the test food group discontinued the study due to the onset of autoimmune pancreatitis during the test food intake period, and one patient in the control food group discontinued the study for personal reasons, so the efficacy analysis set (PPS) was set at 76 patients (38 in the test food group and 38 in the control food group). In addition, two patients in the control food group were excluded from the analysis for only those items due to issues with the reliability of the data for some evaluation items. In one patient, hemolysis was observed in the blood sample, so LDH, K, and insulin in the blood test 4 weeks after intake were excluded from the analysis. In one patient, food intake on the day of the test (non-compliance with management items) was suspected, so blood glucose and insulin in the blood test 8 weeks after intake were excluded from the analysis. The subject background of the subjects in the efficacy analysis is shown in Table 2.

In none of the groups (FAS, PPS, subgroups by BMI, or subgroups by gender) were there any significant differences between the test food group and the control food group in the allocation factors of gender at the time of consent, age, visceral fat area at screening, BMI, waist circumference, or hip circumference.

2. Analysis results (abdominal fat area)
The changes in visceral fat area, subcutaneous fat area, and total fat area in the efficacy analysis set (PPS) are shown in Table 3. No significant difference was observed between the test food group and the control food group in visceral fat area, the primary endpoint. In the test food group, a significant decrease was observed in visceral fat area and total fat area after 12 weeks of intake compared to before intake began.

(Physical measurement items)
Table 4 shows changes in body weight, BMI, body fat mass, muscle mass, body fat mass by region, waist circumference, and hip circumference in the PPS, as well as blood pressure and pulse rate in the FAS. Four weeks after ingestion, the test food group showed significantly higher systolic and diastolic blood pressure values and significantly higher changes in pulse rate compared to the control food group, but no significant changes were observed thereafter. No significant differences were observed between the test food group and the control food group in any of the other evaluation items.

(Blood and urine tests)
Table 5 shows the changes in blood test items (secondary evaluation items) in the PPS. For T-Cho, the test food group showed a significant suppression of the change from before the start of intake compared to the control food group after 4 weeks of intake. For blood glucose, the test food group showed significantly higher values than the control food group before the start of intake, and showed significantly higher values even after 4 weeks of intake, but no difference was observed thereafter. For insulin levels, the test food group showed a significant decrease compared to the control food group. In addition, no clinically significant changes were observed in any of the subjects for other blood test items and urine test items in the FAS.

(Activity monitor)
During the study period, activity levels were checked every two weeks in terms of number of steps, active steps, and duration of moderate intensity activity, but no significant differences were found between the test food and control food groups.

(Subgroup analysis)
The results of the subgroup analysis of BMI before the start of intake from 23 kg/ ^m2 to less than 25 kg/ ^m2 are shown in Table 6. In the subgroup analysis of BMI before the start of intake from 23 kg/ ^m2 to less than 25 kg/ ^m2 , the test food group showed a significantly lower visceral fat area after 12 weeks of intake compared to the control food group in the evaluation item of abdominal fat area. In addition, a significant decrease was observed in the change in subcutaneous fat area and total fat area from before the start of the study after 12 weeks of intake. In addition, in the physical measurement items, the test food group showed a significant decrease in the change in weight and BMI from before the start of intake after 12 weeks of intake compared to the control food group. Furthermore, in the evaluation items of blood tests, the test food group showed a significant increase in the change from before the start of intake for total ketone bodies, acetoacetate, and 3-hydroxybutyrate after 12 weeks of intake compared to the control food group. A significant increase was observed in HDL-Cho after 4, 8, and 12 weeks of intake. For TG, a significant low was observed after 12 weeks of intake, and a significant decrease in the amount of change from before the start of intake was observed. For blood glucose, a significant high was observed before intake, and 4 and 8 weeks after intake. For insulin, a significant increase in the amount of change from before the start of intake was suppressed after 12 weeks of intake.

In the subgroup analysis of those with a BMI of 25 kg/ ^m2 or greater but less than 30 kg/ ^m2 before intake and in the subgroup analysis by gender, no significant differences were observed between the test food group and the control food group in any of the evaluation items.

(Discussion)
In this study, the body fat reducing effect of licorice extract-containing food was examined in healthy adult men and women with a BMI of 23 kg/ ^m2 or more but less than 30 kg/ ^m2 and an abdominal visceral fat area of 100 ^cm2 or more in a randomized placebo-controlled double-blind parallel group comparative study in which a capsule containing 100 mg of licorice extract (test food) or a capsule not containing licorice extract (control food) was taken once a day for 12 weeks. As a result, in the subgroup with a BMI of 23 kg/ ^m2 or more but less than 25 kg/ ^m2 , the test food group showed significantly lower visceral fat area after 12 weeks of intake compared to the control food group, and the changes in subcutaneous fat area and total fat area from before the start of the study, as well as the changes in body weight and BMI from before the start of the study, were significantly reduced, indicating that licorice extract has a body fat reducing effect in humans as well, and associated decreases in body weight and BMI were observed.

Regarding the mechanism of licorice extract's body fat reducing effect, the following test confirmed that it promotes the breakdown of TG in fat cells in cell tests. In animal tests, inhibition of body fat accumulation and reduction in fat cell size were also confirmed, and results that support the results of the cell tests have been reported. A body fat reducing effect was also observed in this test, and significant increases were observed in total ketone bodies, acetoacetic acid, and 3-hydroxybutyric acid in the test food group compared to the control food group. An increase in blood ketone bodies is an indicator of fatty acid consumption through β-oxidation and is also seen in cases of insulin resistance, but no increase in blood glucose or insulin levels was observed, and it was thought that this was not due to an increase in insulin resistance, but rather due to increased energy consumption, as has been reported in other functional foods.

These results suggest that when TGs are broken down in fat cells following ingestion of licorice extract, free fatty acids are released and quickly consumed as an energy source through β-oxidation, resulting in a reduction in body fat.

The tests described below have confirmed that gliasperin B in licorice extract contributes nearly 100% to TG decomposition in fat cells. Based on the above, it is believed that the body fat reducing effect of ingesting licorice extract is due to gliasperin B.

　The safety of licorice extract was also confirmed in this study. As a result, no adverse events attributable to the intake of licorice extract, including abnormalities in clinical test values, were observed, and it was considered that there were no issues with the safety of long-term intake of licorice extract.

(Conclusion)
Licorice extract-containing foods have been shown to have a body fat reducing effect in healthy adult men and women who are slightly obese (BMI 23 kg/ ^m2 or more and less than 25 kg/ ^m2 , abdominal visceral fat area 100 ^cm2 or more). In addition, it is thought that part of the mechanism of action is the decomposition of TG in fat cells and the accompanying enhancement of fatty acid consumption (β-oxidation).

<Test 2>
Five-week-old male C57BL/6J mice were divided into groups of six and fed MF (Oriental Yeast), high-fat, high-carbohydrate powdered feed (hereinafter abbreviated as HFS, D12079BM, Research Diet), or HFS mixed with licorice extract for 8 weeks. After 8 weeks of ingestion, the body weight and visceral fat weight (the sum of epididymal fat weight, perirenal fat weight, and mesenteric fat weight) were measured. The licorice extract was obtained by subjecting the powder obtained by crushing the root and stolon of licorice to a 50% (v/v) ethanol extraction process, drying the liquid by an evaporator and freeze-drying, and then crushing the liquid. These were each mixed with the feed at 0.3% and fed to the mice. The amount of gliasperin B contained in the licorice extract, and the body weight gain and visceral fat weight of the mice are shown in Table 7.

(Conclusion)
Mice that ingested licorice extract containing a higher amount of gliasperin B experienced a slower weight gain and a lower visceral fat weight. This suggests that gliasperin B is closely involved in the body fat/visceral fat reducing effect of licorice extract, and is also involved in the results of Test Example 1.

<Test 3> Preparation of Gliasperin B 10 ml of distilled water was added per 1 g of licorice extract to prepare an aqueous solution of licorice extract. Half the volume of ethyl acetate was added to this aqueous solution, and the solution was mixed to separate into two layers, from which the ethyl acetate layer was collected. This operation was repeated three times. Anhydrous sodium sulfate was added to the obtained ethyl acetate layer, and the mixture was filtered to obtain an organic fraction. The organic fraction was concentrated using an evaporator, and the fraction eluted with hexane:ethyl acetate = 1:1 (v/v) using a silica gel column chromatograph was repeatedly purified using a high-performance liquid chromatograph (acetonitrile:water = 60:40, 45:55 (both v/v)) using an ODS column to obtain a compound (gliasperin B). The gliasperin B (GB) obtained here was used as a test substance in the following test using mouse preadipocytes 3T3-L1.

The above licorice extract was obtained by the following method. That is, the stem, root, and stolons of licorice were crushed to obtain licorice powder. 50 g of licorice powder was extracted with 500 mL of 50% aqueous ethanol at 20°C, and the resulting extract was frozen and dried in an evaporator to obtain a 50% aqueous ethanol extract of licorice.

The spectral data of the compound is shown below, and shows a very good agreement with the spectral data of gliasperin B, which has been reported in the literature as having been isolated from plants of the genus Glycyrrhiza (Zeng L. et al., Heterocycles 34:575-587 (1992)). Since there was no inconsistency with the structural information, the compound was identified as gliasperin B (GB).

1H-NMR (acetone-d) 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 (acetone-d) 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)

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

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

<Test 4> Study of lipolytic effect of licorice extract and gliasperin B (GB) Mouse preadipocytes 3T3-L1 were cultured in differentiation induction medium (0.5 mM isobutyl-methylxanthine, 1 μM dexamethasone, 1 μg/mL insulin in 10% FBS/DMEM) for 2 days, and then the medium was replaced with 10% FBS/DMEM medium containing 1 μg/mL insulin and cultured for 7 days. The test substances (licorice extract and GB) were added on the 9th day after the start of differentiation induction, and the cells were then cultured for 19 hours. The licorice extract used in Test 1 was used as the licorice extract (final concentrations: 100 μg/mL, 200 μg/mL, 300 μg/mL), and the GB prepared in Test 1 was used as the GB (final concentrations: 13.5 ng/mL, 27 ng/mL, 40 ng/mL). Three subjects were tested for each group. Glycerol released into the medium upon decomposition of neutral fats accumulated in the cells was quantified using Labo Assay (TM) Triglyceride (Fujifilm Wako Pure Chemical). The results are shown in Figure 1. The amount of glycerol is shown as the mean value ± standard deviation. The symbols in Figure 1 indicate the following: ** p<0.01 vs. no treatment (Student's t-test).

As shown in Figure 1, the amount of glycerol released increased in a GB concentration-dependent manner, indicating that intracellular lipolysis occurred in a GB concentration-dependent manner, and GB was found to promote lipolysis in adipocytes. On the other hand, licorice extract also released glycerol from adipocytes, but the amount of glycerol released was less when the licorice extract concentration was 300 μg/mL than when it was 100 μg/mL or 200 μg/mL. The concentrations of licorice extract, 100 μg/mL, 200 μg/mL, and 300 μg/mL, respectively, are converted to the amount of GB contained in 13.5 ng/mL, 27 ng/mL, and 40 ng/mL. Licorice extract also has an excellent lipolysis effect, but the lipolysis effect decreased as the concentration increased, suggesting that licorice extract contains various components, some of which may inhibit lipolysis. From this, it can be said that in order to obtain a higher lipolysis effect, it is preferable to use a refined GB product rather than a licorice extract.

<Test 5> Effects of licorice extract on body weight, BMI, and visceral fat level (index value of visceral fat accumulation) A subject (one male, aged 59) took 200 mg of the licorice extract used in Test 1 per day for 100 days. The subject was given a record sheet and asked to record the body weight, BMI, and visceral fat level measured with a Tanita body composition scale before the start of licorice extract intake (day 0), on the 30th day of intake, the 45th day of intake, the 72nd day of intake, the 83rd day of intake, the 92nd day of intake, and the day after the final intake (day 101). The changes in body weight, BMI, and visceral fat level from before the start of intake are shown in Figure 2.

As shown in Figure 2, it was shown that the intake of licorice extract containing GB first reduced visceral fat levels, followed by reductions in body weight and BMI. Considering this together with the results of Test 3 above, it is believed that the GB in the licorice extract efficiently decomposed fat in adipocytes, reducing body fat such as visceral fat.

<Test 6>
Five-week-old male C57BL/6J mice were divided into groups of six and fed MF (Oriental Yeast), high-fat, high-carbohydrate powdered feed (hereinafter abbreviated as HFS, D12079BM, Research Diet), or HFS mixed with licorice extract for 8 weeks. After 8 weeks of ingestion, the visceral fat weight (the sum of the epididymal fat weight, perirenal fat weight, and mesenteric fat weight) was measured. For the licorice extract (Test Examples 1 to 5), the root and stolon of licorice were crushed into powder, and the solution obtained through a 50% (v/v) ethanol extraction process was dried and solidified using an evaporator and freeze-dried, and then crushed to obtain a licorice extract. These were mixed with the feed in the amounts (%) shown in Table 8 below and fed to the mice. The human-equivalent dose of gliasperin (μg/day) and the visceral fat weight suppression rate (%) of the mice are also shown in Table 8.

(Conclusion)
As shown in the above table, a significant visceral fat weight suppressing effect was observed when the administration (ingestion) amount of gliasperin to mice was 9.7 μg/day to 47.6 μg/day calculated in human terms.

The internal composition of the present invention contains gliasperin B, which has an excellent lipolysis promoting effect and a significant weight loss effect. By using the internal composition of the present invention, fat in adipocytes can be efficiently broken down and body fat such as subcutaneous fat and visceral fat can be reduced, resulting in a significant weight loss effect and an improvement in BMI. In addition, the composition improves insulin resistance and reduces sterol levels. Therefore, the internal composition of the present invention can prevent and improve obesity and lifestyle-related diseases in the long term, thereby maintaining and improving health. Furthermore, the content of gliasperin B in a plant extract is an indicator of the effectiveness of the plant extract for lipolysis, weight loss, etc., making it possible to efficiently select a highly effective plant extract.

Claims (10)

An internal composition containing gliasperin B that is used for one or more of the following: decomposing fat, burning fat, assisting in fat consumption, improving BMI, assisting in lowering BMI, reducing abdominal fat, reducing visceral fat, reducing body fat, reducing waist circumference, improving insulin resistance, and reducing LDL cholesterol levels. An internal composition containing an effective amount of gliasperin B, which is used for one or more of the following: decomposing fat, burning fat, assisting in fat consumption, improving BMI, assisting in lowering BMI, reducing abdominal fat, reducing visceral fat, reducing body fat, reducing waist circumference, improving insulin resistance, and reducing LDL cholesterol levels. The internal composition according to claim 2, wherein the effective amount is 10 μg or more per day for an adult. The internal composition according to claim 1 or 2, which is used for adults with a BMI of 23 kg/ ^m2 or more and less than 30 kg/ ^m2 . 3. The internal composition according to claim 1 or 2, which is used for adults with a visceral fat area of 100 ^cm2 or more. The internal composition according to claim 1 or 2, which is taken at a dose of 5 to 100 μg/day, calculated as the total amount of gliasperin B. A method for determining the effectiveness of a plant extract in breaking down fat and/or reducing body weight in a living body, using the gliasperin B content as an indicator. The method for determining the effectiveness of a compound according to claim 7 in decomposing fat and/or reducing body weight, wherein the plant is licorice. An indicator of the quality of a plant extract, the quality being related to the effectiveness in lipolysis and/or weight loss in a living body, and characterized in that the indicator is made of gliasperin B. The indicator according to claim 9, wherein the plant is licorice.

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