Oral preparations for lipolysis and / or weight loss and methods for determining efficacy.
Patent Information
- Authority / Receiving Office
- VN · VN
- Patent Type
- Applications
- Current Assignee / Owner
- ROHTO PHARM CO LTD
- Filing Date
- 2024-08-09
- Publication Date
- 2026-07-01
Smart Images
Figure VN1202602023_0
Abstract
Description
Internal compositions for lipolysis and / or weight loss, methods for determining efficacy, and indicators of plant extract quality
[0001] 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.
[0002] Body fat occurs when excess energy intake relative to energy expenditure accumulates as neutral fat in white adipocytes. 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 pathologies 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 a cosmetic perspective.
[0003] Traditionally, various measures to prevent and improve obesity have been implemented, 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, resulting in failure to improve obesity. Energy restriction can also lead to deficiencies in necessary nutrients, which can be harmful to health. Therefore, it has become increasingly believed that the ideal way to combat obesity is to actively break down accumulated fat and release it as heat energy, thereby maintaining an ideal body weight. Under these circumstances, active research has been conducted in recent years to identify functional ingredients in food materials that have the effect of promoting lipolysis, and many foods and beverages have been proposed.
[0004] 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 excellent lipolysis-promoting activity (Patent Document 1), that at least one of an extract of birch (Betulaceae) and an extract of Kumazasa (Poaceae) promotes lipolysis in systemic or local adipose tissue, thereby improving obesity and preventing its increase (Patent Document 2), and that certain plants such as Juniperus communis or their extracts are useful as pharmaceuticals, foods, or cosmetics that promote the decomposition of neutral fats accumulated in adipose tissue and exhibit slimming effects such as the suppression, prevention, or amelioration of obesity (Patent Document 3). However, the lipolysis and weight loss effects of these plant extracts are currently insufficient. Patent Document 4 also discloses that a composition combining a licorice hydrophobic extract with antioxidants and other ingredients inhibits body fat accumulation, promotes body fat decomposition, and promotes energy production, but the licorice hydrophobic extract alone does not provide sufficient lipolysis-promoting effects.
[0005] Japanese Patent Application Laid-Open No. 2011-074051 Japanese Patent Application Laid-Open No. 2006-045120 Japanese Patent Application Laid-Open No. 2012-229266 International Publication No. 2008 / 143182
[0006] The problem to be solved by the present invention is to provide a novel preparation that is excellent in lipolysis promoting effect, visceral fat / body fat reducing effect, weight loss effect, and BMI improving effect. Another object of the present invention is to provide a novel preparation that is excellent in insulin resistance improving effect, cholesterol level reducing effect, etc.
[0007] As a result of intensive research, the present inventors have found that "gliasperin B," a specific component in licorice extract, has excellent lipolysis-promoting properties. They have also found that a preparation containing licorice extract containing gliasperin B not only exhibits excellent lipolysis-promoting properties, but also has the effects of reducing visceral fat and body fat, reducing body weight, improving BMI, improving insulin resistance, and reducing cholesterol levels. Furthermore, they have found that the content of gliasperin B in a plant extract serves as an indicator of the effectiveness of the plant extract for lipolysis, weight loss, etc. The gist of the present invention is as follows:
[0008] [1] An internal composition containing gliasperin B, which is used for one or more of fat decomposition, fat burning, assisting 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, which is used for one or more of fat decomposition, fat burning, assisting 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 internal composition according to [2], wherein the effective amount is 10 μg or more per day for an adult. [4] A composition for a patient with a BMI of 23 kg / m ２ More than 30kg / m ２ [5] The oral composition according to [1] or [2], which is used for adults with a visceral fat area of less than 100 cm. ２ The oral composition according to [1] or [2], which is used for the above-mentioned adults. [6] The oral composition according to [1] or [2], which is ingested at a dose of 5 to 100 μg / day, calculated as the total amount of gliasperin B. [7] A method for determining the effectiveness of a plant extract on lipolysis and / or weight loss in a living body, using the content of gliasperin B as an index. [8] The method for determining the effectiveness on lipolysis and / or weight loss according to [7], wherein the plant is licorice. [9] An index of the quality of a plant extract, wherein the quality is related to the effectiveness on lipolysis and / or weight loss in a living body, and is characterized in that the index consists of gliasperin B.
[10] The index according to [9], wherein the plant is licorice.
[0009] The internal composition of the present invention contains gliasperin B, preferably as an active ingredient, functional ingredient, indicator ingredient, or contributing ingredient, and thereby exhibits excellent lipolysis-promoting activity and a significant weight loss effect. Use of the internal composition of the present invention efficiently decomposes fat in adipocytes and reduces body fat, such as subcutaneous fat and visceral fat, resulting in significant weight loss and improved BMI. It also improves insulin resistance and reduces sterol levels. Thus, the internal composition of the present invention can prevent and improve obesity and lifestyle-related diseases over the long term, thereby maintaining and improving health. Furthermore, the gliasperin B content in a plant extract serves as an indicator of the plant extract's effectiveness in lipolysis, weight loss, and other areas, enabling efficient selection of highly effective plant extracts.
[0010] Figure 1 shows the lipolysis promoting effect of licorice extract and gliasperin B. Figure 2 shows the effect of licorice extract on body weight, BMI and visceral fat level.
[0011] The internal composition of the present invention, the method for determining the effectiveness of lipolysis and / or weight loss, and the indicators of the quality of the plant extract are described in detail below.
[0012] <Internal Composition> 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, functional ingredient, indicator ingredient, or contributing ingredient. In addition to gliasperin B, which is an essential ingredient, the internal composition of the present invention may contain other ingredients to the extent that the effects of the present invention are not impaired. Below, gliasperin B and other ingredients contained in the internal composition of the present invention, as well as the form of the internal composition of the present invention, etc. will be explained.
[0013] In this specification, the term "active ingredient" refers to a pharmacologically active ingredient or physiologically active ingredient contained in a drug, quasi-drug, etc. that exhibits the intended effect.
[0014] In this specification, functional ingredients refer to ingredients contained in internal compositions such as foods that contribute to specific health purposes. 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 are ingredients that can be directly or indirectly confirmed qualitatively and quantitatively. Functional foods are provided to consumers with a display of the functions based on the effects of the ingredients contained in the food. When providing a food with functional claims to consumers, the content of the functional ingredients and the method for quantifying them must be notified.
[0015] In this specification, the term "index component" refers to an extract, essence, etc. that is considered to be a functional component when specific components that can explain part of the scientific basis of the functionality have been identified, but the functionality cannot be fully explained by these specific components alone, and the mechanism of action related to the functionality to be displayed has been considered for at least one component through in vitro tests, in vivo tests, or clinical trials (human tests).
[0016] In this specification, a contributing component 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 component could be, but is not limited to, a "contributing component," a "functionally contributing component," or an "indicator component." In pharmaceuticals and quasi-drugs, the equivalent of a contributing component is an "active ingredient." Furthermore, in the sale of food compositions other than foods with functional claims, the 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."
[0017] (Glyasperin B) Glyasperin B (hereinafter also referred to as "GB") is an isoflavanone derivative represented by the following formula: 3-(2,4-dihydroxyphenyl)-5-hydroxy-2,3-dihydro-6-(3-methyl-2-butenyl)-7-methoxy-4H-1-benzopyran-4-one, 3-(2,4-dihydroxyphenyl)-5-hydroxy-6-(3-methyl-2-butenyl)-7-methoxy-3,4-dihydro-2H-1-benzopyran-4-one, or 3-(2,4-dihydroxyphenyl)-5-hydroxy-7-methoxy-6-(3-methylbut-2-en-1-yl)-3,4-dihydro-2H-1-benzopyran-4-one.
[0018] Gliasperin B is a component confirmed to be contained in licorice extract, and the gliasperin B of 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 abundant in licorice extract, may also have a lipolysis-promoting effect (see Patent Document 4, etc.). However, it was unclear which component in licorice extract has the lipolysis-promoting effect. The present inventors were the first to discover that, among the many components present in licorice extract, gliasperin B has a lipolysis-promoting effect. They have then completed the invention of an internal composition containing gliasperin B, which has the effects of fat decomposition, fat burning, fat consumption support, BMI improvement, BMI reduction support, abdominal fat reduction, visceral fat reduction, body fat reduction, waist circumference reduction, insulin resistance improvement, and LDL cholesterol reduction.
[0019] Gliasperin B in the present invention may be derived from licorice or may be a chemically synthesized product. In this specification, "containing gliasperin B" includes the case of containing a plant such as licorice that contains gliasperin B.
[0020] The type of licorice is not particularly limited, and examples thereof include plants of the genus Glycyrrhiza in the family Fabaceae, such as Glycyrrhiza uralensis (G. uralensis Fisch. et DC; ural licorice), Glycyrrhiza inflata (G. inflata BAT.; chinese licorice), Glycyrrhiza glabra (G. glabra L.; day licorice), Glycyrrhiza glabra (G. glabra L.var glandu lifera Regel et Herder; tallow tree licorice), Glycyrrhiza aspera, Glycyrrhiza echinata (G. echinata L.; Chinese licorice), Glycyrrhiza pallidiflora (G. pallidiflora Maxim; Glycyrrhiza glabra) and the like.
[0021] An example of a method for obtaining gliasperin B derived from licorice is a method for purifying it from a licorice extract. Specifically, the licorice extract is a licorice extract obtained by extracting the whole licorice plant or a pulverized product of a part of the plant (e.g., root, stem, stolon, leaf, flower, fruit, etc.) with a solvent, and the resulting 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 with alcohols or ketones such as acetone. Of these, water, alcohol, and aqueous alcohol are preferred as the extraction solvent, with hot water, ethanol, and aqueous ethanol being more preferred.
[0022] The alcohol concentration of the hydrous alcohol is 0.1% by mass to 99.9% by mass, preferably 10% by mass to 99.9% by mass, more preferably 30% by mass to 70% by mass, even more preferably 40% by mass to 60% by mass, and particularly preferably 50% by mass, or 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).
[0023] The dried licorice extract is dissolved in distilled water to prepare an aqueous licorice extract solution. An organic solvent such as ethyl acetate is added in an amount equal to half the volume of the aqueous solution, and the mixture is mixed to separate into two layers. The organic solvent layer is then recovered, and this process is repeated multiple times, preferably three times. Anhydrous sodium sulfate is added to the resulting organic solvent layer, followed by filtration to obtain an organic fraction. The organic fraction is concentrated using an evaporator, and the fraction eluted with a hexane:ethyl acetate ratio of 1:1 (v / v) or similar mixture is purified by high-performance liquid chromatography using an ODS column (acetonitrile:water ratios of 60:40 and 45:55 (v / v)) to obtain the compound (gliasperin B).
[0024] The content of gliasperin B in the internal composition of the present invention is 0.00001% by mass to 50% by mass, preferably 0.0001% by mass to 20% by mass, more preferably 0.0005% by mass to 10% by mass, and even more preferably 0.001% by mass to 5% by mass.
[0025] (Other Components) In addition to gliasperin B, the oral composition of the present invention may contain carriers, excipients, solvents, and other optional components to the extent that the effects of the present invention are not impaired.
[0026] 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 falls within the above-mentioned numerical range. Furthermore, from the viewpoint of promoting lipolysis, the internal composition of the present invention preferably contains gliasperin B within the above-mentioned 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 index of licorice extract in the Japanese Pharmacopoeia, as a comparison standard.
[0027] 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 pharmaceutical or a cosmetic agent, etc., to efficiently break down fat and reduce body fat such as subcutaneous fat and visceral fat.
[0028] 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, color formers, bleaching agents, antifungal agents (mold inhibitors), yeast food, gum base, flavorings, acidulants, seasonings, emulsifiers, pH adjusters, alkaline water, leavening agents, nutritional supplements, and other food and beverage ingredients may be mixed to prepare the composition into the desired form. When preparing the internal composition of the present invention in the form of a food or beverage, the form is not particularly limited. Examples include supplement-type foods such as gels, granules, fine granules, capsules, tablets, powders, liquids, and semisolids; beverages such as carbonated drinks, soft drinks, dairy drinks, alcoholic drinks, fruit juice drinks, teas, and nutritional drinks; powdered beverages such as powdered juice and powdered soup; confectioneries such as gums, tablets, candies, cookies, gummies, rice crackers, biscuits, and jellies; bread, noodles, cereals, jams, and seasonings. These foods are used as foods and beverages for efficiently breaking down fat and reducing body fat such as subcutaneous fat and visceral fat, and can be used, for example, as nutraceuticals such as general foods and beverages, nutritional supplements, functional foods, foods for specified health uses, and foods for the sick.
[0029] When the internal composition of the present invention is prepared as a pharmaceutical product (including quasi-drugs), other active ingredients, pharmaceutically acceptable carriers, additives, etc. may be optionally blended in addition to gliasperin B, as needed. Specific examples of pharmaceutically acceptable carriers and additives include binders, disintegrants, lubricants, humectants, buffers, preservatives, and flavorings. When the internal composition of the present invention is prepared as a pharmaceutical product, its form is not particularly limited. Examples include injections, topical 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 pharmaceutical products) 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 such as subcutaneous fat and visceral fat.
[0030] 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 pharmaceutically or cosmetically acceptable carrier (e.g., water, oily components) is added to gliasperin B and formulated into the desired form. The cosmetic is not particularly limited in its form, as long as it is applicable to the skin. Examples of forms include liquid, emulsion, powder, solid, suspension, cream, ointment, mousse, granule, tablet, gel, jelly, paste, aerosol, spray, liniment, and pack. These cosmetic preparations are used to efficiently decompose fat and reduce subcutaneous fat.
[0031] The dosage and application amount of the internal composition of the present invention can be determined appropriately 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, or 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 daily dose of gliasperin B ingested 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, still 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, calculated as the total amount of gliasperin B.
[0032] 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. As used herein, "effective amount" refers to the amount administered when the desired effect is observed, whether through direct or indirect action. The effective amount varies depending on age and body weight, but may 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 of the total amount of gliasperin B described above per day for an adult, 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 gliaperin 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, still 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.
[0033] The oral composition of the present invention is suitable for patients with a BMI of 23 kg / m or less, from the viewpoint of easily achieving a weight loss effect. ２ More than 30kg / m２ It is preferably used for adults with a BMI of less than 23 kg / m ２ More than 25kg / m ２ According to the criteria of the Japan Society for the Study of Obesity, it is more preferable to use it for adults with a BMI of 18.5 kg / m or less. ２ More than 25kg / m ２ Less than 25 kg / m is considered normal weight. ２ More than 30kg / m ２ An obesity level of less than 1 is considered to be an obesity level. The oral composition of the present invention is preferably used for adults who are so-called normal weight to slightly obese (obesity level 1), and is particularly preferably used for adults who are not classified as obese but are slightly obese within a normal weight range. BMI (Body Mass Index) is a body mass index that indicates the degree of obesity, calculated from weight and height using the following formula. BMI is used as an international index for adults. Formula: BMI = weight (kg) / (height (m)) ２
[0034] From the viewpoint of easily achieving the lipolysis promoting effect and weight loss effect, the internal use composition of the present invention is ２ The visceral fat area can be calculated in the usual way by taking a CT scan of the abdomen and taking a plurality of slice images at intervals of 1 cm, centered on the navel.
[0035] 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 lowering cholesterol levels, which contains gliasperin B as an active ingredient.
[0036] The present invention also includes the above-mentioned weight loss agents, visceral fat / body fat reducing agents, BMI improving agents, insulin resistance improving agents, and cholesterol level reducing agents, which, by containing gliasperin B, have excellent lipolysis promoting activity and exhibit significant weight loss effects. By using the weight loss agents, visceral fat / body fat reducing agents, BMI improving agents, insulin resistance improving agents, and cholesterol level reducing agents 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 significant weight loss and BMI improvement effects. Insulin resistance improvement and sterol level reduction effects can also be achieved. Therefore, the weight loss agents, visceral fat / body fat reducing agents, BMI improving agents, insulin resistance improving agents, and cholesterol level reducing agents of the present invention can prevent and improve obesity and lifestyle-related diseases over the long term, thereby maintaining and improving 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 directly applied to the explanation in the section on the internal composition of the present invention.
[0037] <Method for Determining the Effectiveness of a Plant Extract for Lipolysis and / or Weight Loss> The present invention also encompasses a method for determining the effectiveness of a plant extract for lipolysis and / or weight loss in vivo using the content of gliasperin B as an indicator. As described above, preparations containing gliasperin B as an active ingredient have excellent lipolysis-promoting properties and exhibit significant weight loss effects. Furthermore, they can efficiently decompose fat in adipocytes and reduce body fat, such as subcutaneous fat and visceral fat, resulting in significant weight loss and improved BMI. They also improve insulin resistance and reduce sterol levels. Since the above effects increase with the content of gliasperin B, using the content of gliasperin B in a plant extract as an indicator makes it possible to determine the effectiveness of the plant extract for lipolysis and / or weight loss in vivo. The plant extract is not particularly limited as long as it contains gliasperin B, but licorice extract is preferred.
[0038] 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 a plant extract (hereinafter also referred to as the "measurement step"). Preferably, the method also 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").
[0039] The method for measuring the gliasperin B content in the plant extract in the measurement step is not particularly limited, and can be determined by methods known to those skilled in the art. Examples include Western blotting and ELISA. In both Western blotting and ELISA, the gliasperin B content can be measured with high accuracy using an anti-gliasperin B antibody that specifically binds to gliasperin B. In Western blotting, the gliasperin B content in the sample can be calculated by comparing the intensity of the resulting gliasperin B-specific blot with the intensity of a standard sample (positive control) of known concentration.
[0040] Gliasperin B used as a standard in the determination method of the present invention can be obtained, for example, by extraction from natural sources, expression of a recombinant nucleic acid encoding gliasperin B, or chemical synthesis.
[0041] In the determination 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 a standard gliasperin B. The standard amount of gliasperin B can be selected appropriately depending on the purpose and subject of evaluation.
[0042] <Indicator of Plant Extract Quality> The present invention also includes an invention relating to an indicator of the quality of a plant extract, the quality being related to the effectiveness of the indicator in lipolysis and / or weight loss in the body, characterized in that the indicator comprises gliasperin B. As described above, preparations containing gliasperin B as an active ingredient have excellent lipolysis-promoting activity and exhibit significant weight loss effects. Furthermore, they can efficiently decompose fat in adipocytes and reduce body fat, such as subcutaneous fat and visceral fat, resulting in significant weight loss and improved BMI. They also improve insulin resistance and reduce sterol levels. Since the above effects increase with the content of gliasperin B, using gliasperin B in a plant extract as an indicator makes it possible to determine the effectiveness of the plant extract in lipolysis and / or weight loss in the body. Licorice is preferred as the plant in the plant extract.
[0043] 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 a 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.
[0044] The present invention will be explained in more detail below by showing examples, but the scope of the present invention is not limited to these examples.
[0045] As shown below, a randomized, double-blind, parallel-group comparative study was conducted in which subjects took the test food (a hard capsule formulation containing licorice extract) for 12 weeks to evaluate the effect on visceral fat, using a control food (hard capsules not containing licorice extract) as a control.
[0046] <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 color. 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 food. The indistinguishability of the test food and packaging was reviewed and approved by the IRB. The nutritional components and related components of the test food are shown in Table 1 below. The amount of gliasperin B contained in the test food was 14 μg per recommended daily intake.
[0047]
[0048] 2. Subjects The target number of subjects in this study was calculated to be 34 per group, for a total of 68 subjects, assuming a large effect size (0.8) for the test food in a two-group parallel comparison study, a significance level of 0.05, and a statistical power of 80-90%, requiring the inclusion of 26-35 subjects per group. Taking into consideration the possibility of subjects dropping out or being discontinued during the study, the number of subjects enrolled was set at 80 (40 per group). Subjects were recruited from volunteers registered with the contracted testing institution, and 81 subjects (50 men, 31 women) who met the following inclusion criteria, did not violate any of the exclusion criteria, and were deemed appropriate for participation by the principal investigator were enrolled as subjects.
[0049] (Inclusion criteria) In the screening test, participants were required to meet the following conditions: (1) men and women aged 20 to 65 years, and (2) a BMI of 23.0 kg / m ２ 30.0kg / m or more ２ (3) Those with a visceral fat area of less than 100 cm ２(4) Those who have received a full explanation of the purpose and contents of this study, are capable of consent, fully understand the purpose and contents, and voluntarily volunteer to participate and can provide written consent to participate in this study. (Exclusion criteria) (1) Those suffering from serious cardiovascular disorders, liver dysfunction, renal dysfunction, respiratory disorders, endocrine disorders, or metabolic disorders, or those with a history of these, (2) Those receiving treatment for chronic diseases such as dyslipidemia, hypertension, or diabetes, (3) Those with psychiatric disorders such as depression, schizophrenia, or bulimia nervosa, (4) Those 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) Those who regularly use supplements or health foods (foods for specified health uses, foods with functional claims, etc.) that may affect the evaluation of this study, such as body fat, triglycerides, body weight, cholesterol, and energy metabolism, (6) Those who may be allergic to the test foods, (7) Persons with metal in the abdominal CT scan area, (8) Persons with implanted medical devices such as cardiac pacemakers or implantable cardioverter defibrillators, (9) Persons with claustrophobia, (10) Persons with smoking habits, (11) Persons with the habit of drinking large amounts of alcohol (60g 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 5 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 swelling 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 who are 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.
[0050] 3. Study Protocol This study was a randomized, placebo-controlled, double-blind, parallel-group comparative study. Tests were conducted at the start of test food intake, and after 4, 8, and 12 weeks of intake. For the subjects enrolled by the study director, a randomization manager, who was not involved in the implementation of the study, generated random numbers to randomize the subjects. At this time, it was confirmed that there were no significant differences between groups in the allocation factors of gender and age at the time of consent acquisition, and visceral fat area, BMI, hip circumference, and waist circumference at the time of screening examination.
[0051] 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.
[0052] During the study period, subjects were instructed to maintain their usual dietary habits as much as possible, and were prohibited from consuming health foods (e.g., foods for specified health uses, foods with functional claims, etc.) and grapefruit juice, which may affect the study's evaluations of body fat, triglycerides, body weight, cholesterol, and energy metabolism. Normal alcohol intake was also maintained, and any amount exceeding the normal intake was prohibited. Alcohol consumption was prohibited the day before the test, and all intake other than water was prohibited after 9:00 PM. Furthermore, all intake other than water was prohibited four hours before abdominal CT. During the study period, subjects were instructed to maintain their usual exercise habits (frequency, type of exercise, etc.) as much as possible, and were prohibited from changing their exercise habits. Except in emergencies, the use of new medications during the study period required permission from the study physician. Vaccinations were prohibited for two weeks prior to the test. Blood donation was prohibited during the study period.
[0053] This study was conducted in accordance with the spirit of the Declaration of Helsinki (2013)—Tokyo, Venice, Hong Kong, Somerset West, Edinburgh (amended), Washington, Tokyo (annotated), Seoul (amended), and Fortaleza (amended)—and in accordance with the "Ethical Guidelines for Life Science and Medical Research Involving Human Subjects (established March 23, 2021, enforced June 30, 2021, and partially revised March 10, 2022)." The study was approved by the Institutional Review Board (IRB) of the Fukuda Internal Medicine Clinic, Koseikai Medical Corporation (approval number: IRB-20220716-2). The study was also registered with the University Hospital Medical Information Network Clinical Trial Registry (UMIN) (UMIN study ID: UMIN000048430).
[0054] 4. Examination items 1) Physical examination and abdominal CT scan Physical examination items included height (only at the time of screening), body composition assessment using InBody770 (InBody Japan Co., Ltd.) (weight, BMI, body fat percentage, muscle mass, regional body fat mass, basal metabolic rate, other body composition component analysis, etc.), waist circumference, hip circumference, blood pressure, and pulse rate, and those who met the selection criteria underwent CT scans. Weight was measured using a DST-210N (Muratec KDS Co., Ltd.) 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 the start of intake.
[0055] Abdominal CT scans were performed using a Discovery 710 or Discovery MIDR (GE Healthcare Japan Co., Ltd.), and three slices were taken at 1 cm intervals centered on the umbilicus. 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 umbilicus position, a second scan was performed centered on the fourth lumbar vertebra.
[0056] 2) Food and activity records During the test period, subjects were asked to record alcohol intake, medication 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, for the three days prior to the test, subjects were asked to record the amount of food consumed as well as the content of their meals. 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 status 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.
[0057] 3) Fatigue and sleep evaluation Fatigue was evaluated by VAS at the start of intake and after 4, 8, and 12 weeks of intake at the time of waking at home. Sleep was evaluated by the OSA sleep questionnaire (MA version) at the start of intake and after 4, 8, and 12 weeks of intake at the time of waking at home.
[0058] 4) Blood and urine tests Blood samples were 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), triglycerides (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 ingestion.
[0059] 5. Statistical analysis The primary endpoint was visceral fat area measured by abdominal CT. Secondary endpoints were body weight, BMI, subcutaneous fat area measured by abdominal CT, total fat area, body fat percentage measured by InBody 770, body fat mass, muscle mass, regional body fat mass, basal metabolic rate, waist circumference, hip circumference, waist-to-hip ratio, VAS (fatigue), OSA Sleep Questionnaire MA version, ketone body fractions measured by blood test, T-Cho, LDL-Cho, HDL-Cho, TG, blood glucose, HbA1c, insulin, and glycoalbumin.
[0060] Statistical analysis was performed using the statistical processing software SPSS Ver. 26 (IBM Japan, Ltd.), with a significance level of 5% for a two-sided test. In addition, to confirm the tendency according to the subject's attributes, the subjects with a BMI of 23 kg / m before the start of intake were included. ２ More than 25kg / m ２ Those with a BMI of less than 25 kg / m ２ More than 30kg / m ２ Subgroup analyses were conducted by those under 18 years of age and by gender. Mean values and standard deviations for each test period and the change from before intake to each test period were tabulated by group, and an unpaired t-test was performed on the difference in mean values between the test food group and the control food group at each test period to compare the differences between groups. Dunnett's multiple comparison test was also performed on the difference in mean values between before intake and each test period for each test food group to compare the differences due to the duration of test food intake. For abdominal CT scans, a paired t-test was performed to compare the differences due to the duration of test food intake. Note that no multiplicity correction was performed for the paired t-test.
[0061] (Results) 1. Subjects analyzed A total of 81 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 exclusion criterion (2), and the study began with 80 subjects. 78 subjects (39 in the test food group, 39 in the control food group) were included in the safety analysis set (FAS), excluding one subject in the test food group who was found to violate 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. 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, resulting in a efficacy analysis set (PPS) of 76 patients (38 in the test food group, 38 in the control food group). Furthermore, two patients in the control food group were excluded from the analysis due to issues with the reliability of data on some evaluation items. One patient had hemolysis in the blood sample, so LDH, K, and insulin in the blood test four weeks after intake were excluded from the analysis. One patient was suspected of having a dietary intake on the day of the test (non-compliance with the management instructions), so blood glucose and insulin in the blood test eight weeks after intake were excluded from the analysis. The subject background of the efficacy analysis subjects is shown in Table 2.
[0062]
[0063] In all subgroups, including FAS, PPS, BMI subgroups, and gender subgroups, no significant differences were observed 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, and hip circumference.
[0064] 2. Analysis results (abdominal fat area) 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 for visceral fat area, the primary endpoint. Regarding changes over time within 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.
[0065]
[0066] (Physical measurement items) Table 4 shows changes in weight, BMI, body fat mass, muscle mass, regional body fat mass, waist circumference, and hip circumference in the PPS, as well as blood pressure and pulse rate in the FAS. With regard to blood pressure and pulse rate, the test food group showed significantly higher systolic and diastolic blood pressure values and significantly higher change in pulse rate after 4 weeks of ingestion 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.
[0067]
[0068]
[0069] (Blood and urine tests) Changes in blood test items (secondary endpoints) in the PPS are shown in Table 5. For T-Cho, after 4 weeks of intake, the test food group showed a significant suppression of the change from before the start of intake compared to the control food group. For blood glucose, the test food group showed significantly higher values compared to the control food group before the start of intake, and still showed significantly higher values after 4 weeks of intake, but no differences were observed thereafter. For insulin levels, the test food group showed a significant decrease compared to the control food group. Furthermore, no clinically significant changes were observed in any of the subjects for other blood test items and urine test items in the FAS.
[0070]
[0071] (Activity Monitor) During the test period, the number of steps, number of active steps, and duration of moderate-intensity activity were checked every two weeks, but no significant differences were observed between the test food and control food groups.
[0072] (Subgroup analysis) BMI before intake was 23 kg / m ２ More than 25kg / m ２ The results of the subgroup analysis are shown in Table 6. ２ More than 25kg / m ２In a subgroup analysis of abdominal fat area, the test food group showed significantly lower visceral fat area values after 12 weeks of intake compared to the control food group. Furthermore, significant decreases were observed in the changes from pre-intake in subcutaneous fat area and total fat area after 12 weeks of intake. Furthermore, in terms of anthropometric measurements, the test food group showed significantly lower changes from pre-intake in weight and BMI after 12 weeks of intake compared to the control food group. Furthermore, in terms of blood test evaluations, significant increases in the changes from pre-intake in total ketone bodies, acetoacetate, and 3-hydroxybutyrate after 12 weeks of intake compared to the control food group were observed. Significant increases in HDL-Cho were observed after 4, 8, and 12 weeks of intake. Significant decreases in TG were observed after 12 weeks of intake, and a significant decrease in the changes from pre-intake. Significant increases were observed in blood glucose levels before intake and after 4 and 8 weeks of intake. Significant suppression of the increase in insulin after 12 weeks of intake was observed.
[0073]
[0074]
[0075] BMI before intake was 25 kg / m ２ More than 30kg / m ２ In the subgroup analyses by age and sex, no significant differences were observed between the test food group and the control food group in any of the evaluation items.
[0076] (Discussion) In this study, the body fat reducing effect of foods containing licorice extract was evaluated in subjects with a BMI of 23 kg / m ２ More than 30kg / m ２ Less than 100cm of abdominal visceral fat ２ A randomized, placebo-controlled, double-blind, parallel-group comparative study was conducted in which the above healthy adult males and females were asked to take one capsule containing 100 mg of licorice extract (test food) or one capsule not containing licorice extract (control food) once a day for 12 weeks. ２ More than 25kg / m ２In the subgroup of subjects under 18 years of age, the test food group had significantly lower visceral fat area after 12 weeks of intake compared to the control food group, and there were significant decreases in the changes from before the start of the test in subcutaneous fat area and total fat area, as well as in body weight and BMI, indicating that licorice extract also has a body fat-reducing effect in humans, with accompanying decreases in body weight and BMI.
[0077] Regarding the mechanism of the body fat-reducing effect of licorice extract, cell testing confirmed that it promotes the breakdown of TG in adipocytes, as described below. Animal testing also confirmed the suppression of body fat accumulation and a reduction in adipocyte size, supporting the results of the cell testing. This test also demonstrated a body fat-reducing effect, with significant increases in total ketone bodies, acetoacetic acid, and 3-hydroxybutyric acid in the test food group compared with the control food group. Increases in blood ketone bodies are an indicator of fatty acid consumption through beta-oxidation and are also seen in patients with insulin resistance. However, no increases in blood glucose or insulin levels were observed, suggesting that this is not due to an exacerbation of insulin resistance, but rather to increased energy consumption, as has been reported with other functional foods.
[0078] These results suggest that when TGs are broken down in fat cells following ingestion of licorice extract, free fatty acids are released and are quickly consumed as an energy source through beta-oxidation, resulting in a body fat reduction effect.
[0079] The tests described below have confirmed that gliasperin B in licorice extract contributes nearly 100% to TG degradation in adipocytes. Based on the above, it is believed that the body fat reducing effect of licorice extract intake is due to gliasperin B.
[0080] This study also confirmed the safety of licorice extract. As a result, no adverse events attributable to licorice extract intake, including abnormalities in clinical test values, were observed, and it was considered that there are no safety issues with long-term intake of licorice extract.
[0081] (Conclusion) Food containing licorice extract is effective for people with a tendency toward obesity (BMI of 23 kg / m２ Above, 25kg / m ２ Abdominal visceral fat area is less than 100 cm ２ The results showed that sucralose has a body fat-reducing effect in healthy adult men and women. Furthermore, it is thought that part of the mechanism of action is the degradation of TG in adipocytes, which in turn promotes fatty acid consumption (β-oxidation).
[0082] <Study 2> Five-week-old male C57BL / 6J mice were divided into groups of six and fed MF (Oriental Yeast), a high-fat, high-carbohydrate powdered diet (hereinafter abbreviated as HFS, D12079BM, Research Diet), or HFS mixed with licorice extract for 8 weeks. After 8 weeks of feeding, 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 prepared by grinding the root and stolon of licorice into powder, extracting the resulting solution with 50% (v / v) ethanol, evaporating, and freeze-drying it, and then grinding it. Each of these extracts was mixed with the diet at 0.3% and given to the mice. The amount of gliasperin B contained in the licorice extract, as well as the body weight gain and visceral fat weight of the mice, are shown in Table 7.
[0083]
[0084] (Conclusion) Mice that ingested licorice extract with a higher content of gliasperin B experienced a slower weight gain and a lower visceral fat weight. This suggests that gliasperin B is closely related to the body fat and visceral fat reducing effect of licorice extract, and is therefore also related to the results of Test Example 1.
[0085] <Test 3> Preparation of Gliasperin B A licorice extract aqueous solution was prepared by adding 10 ml of distilled water per 1 g of licorice extract. Ethyl acetate was added in an amount equal to half the volume of this aqueous solution, and the mixture was mixed to separate into two layers. The ethyl acetate layer was then recovered. This procedure was repeated three times. Anhydrous sodium sulfate was added to the resulting ethyl acetate layer, followed by filtration 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 high-performance liquid chromatography (acetonitrile:water = 60:40, 45:55 (both v / v)) using an ODS column to obtain the compound (gliasperin B). The resulting gliasperin B (GB) was used as the test substance in the following test using mouse preadipocytes 3T3-L1.
[0086] The licorice extract was obtained by the following method: The stems, roots, and stolons of licorice were crushed to obtain licorice powder. 50 g of the 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.
[0087] The spectral data of the compound is shown below, and shows very good agreement with the spectral data of gliasperin B, which has been reported to be isolated from plants of the genus Glycyrrhiza in the literature (Zeng L. et al., Heterocycles 34:575-587 (1992)). Since no inconsistency with the structural information was observed, the compound was identified as gliasperin B (GB).
[0088] 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. 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)
[0089] The amount of gliasperin B (GB) contained in the licorice extract was quantified by high performance liquid chromatography, as described in detail below. The result showed that the gliasperin B (GB) content was 132.6 μg per gram of licorice extract. The amount of glycyrrhizic acid contained in the licorice extract was 123 mg per gram of licorice extract, and the ratio of the gliasperin B (GB) content to the glycyrrhizic acid content was 0.00107.
[0090] (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: 1 ml / min Detection wavelength: 290 nm GB retention time: 33.2 minutes
[0091] <Test 4> Examination of the 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, then the medium was changed to 10% FBS / DMEM 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 per group. Glycerol, released into the medium upon decomposition of neutral fats accumulated in the cells, was quantified using Laboassay™ Triglyceride (Fujifilm Wako Pure Chemical Industries). The results are shown in Figure 1. The amount of glycerol is shown as the mean ± standard deviation. The symbols in Figure 1 indicate the following: ** p<0.01 vs. untreated (Student's t-test).
[0092] 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 it was found that GB has the effect of promoting lipolysis in adipocytes. Meanwhile, licorice extract also caused glycerol release from adipocytes, but the amount of glycerol released was lower when the licorice extract concentration was 300 μg / mL than when it was 100 μg / mL or 200 μg / mL. Note that the concentrations of licorice extract, 100 μg / mL, 200 μg / mL, and 300 μg / mL, respectively, are converted to the amount of GB contained, 13.5 ng / mL, 27 ng / mL, and 40 ng / mL, respectively. Although licorice extract also has an excellent lipolytic effect, the lipolytic effect decreases as the concentration increases, 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 lipolytic effect, it is preferable to use a purified GB product rather than a licorice extract.
[0093] <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 body weight, BMI, and visceral fat level measured with a Tanita body composition scale before starting licorice extract intake (day 0), on the 30th, 45th, 72nd, 83rd, and 92nd days, and the day after the final intake (day 101). The changes in body weight, BMI, and visceral fat level from before starting intake are shown in Figure 2.
[0094] 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 GB in the licorice extract efficiently decomposed fat in adipocytes, resulting in a reduction in body fat such as visceral fat.
[0095] <Study 6> Five-week-old male C57BL / 6J mice were divided into groups of six and fed MF (Oriental Yeast), a high-fat, high-carbohydrate powdered diet (hereinafter abbreviated as HFS, D12079BM, Research Diet), or HFS mixed with licorice extract for 8 weeks. After 8 weeks of feeding, visceral fat weight (the sum of epididymal fat weight, perirenal fat weight, and mesenteric fat weight) was measured. For the licorice extract (Test Examples 1 to 5), licorice root and stolon were crushed into powder, and the resulting solution was extracted with 50% (v / v) ethanol. The extract was then evaporated and freeze-dried, and then crushed to obtain a licorice extract. Each extract was mixed with the diet in the amounts (%) shown in Table 8 below. The human-equivalent gliasperin dose (μg / day) and the visceral fat weight suppression rate (%) in mice are also shown in Table 8.
[0096]
[0097] (Conclusion) As shown in the table above, a significant visceral fat weight suppressing effect was observed when the dose (intake) of gliasperin administered to mice was 9.7 μg / day to 47.6 μg / day in human terms.
[0098] The internal composition of the present invention contains gliasperin B, which exhibits excellent lipolysis-promoting activity and a significant weight loss effect. Use of the internal composition of the present invention efficiently decomposes fat in adipocytes and reduces body fat, such as subcutaneous fat and visceral fat, resulting in significant weight loss and improved BMI. It also improves insulin resistance and reduces sterol levels. Therefore, the internal composition of the present invention can prevent and improve obesity and lifestyle-related diseases over the long term, thereby maintaining and improving health. Furthermore, the gliasperin B content in a plant extract serves as an indicator of the plant extract's effectiveness in lipolysis, weight loss, and other areas, making it possible to efficiently select highly effective plant extracts.
Claims
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 internal composition according to claim 2, wherein the effective amount is 10 μg or more per day for an adult.
4. BMI is 23kg / m ２ More than 30kg / m ２ The internal composition according to claim 1 or 2, which is used for adults under 6 years of age.
5. Visceral fat area is 100 cm ２ The internal composition according to claim 1 or 2, which is used for adults of the abovementioned age.
6. The internal composition according to claim 1 or 2, which is ingested 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 in breaking down fat and / or reducing body weight 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 claim 7, wherein the plant is licorice.
9. An indicator of the quality of a plant extract, said quality being related to its effectiveness in lipolysis and / or weight loss in the living body, characterized in that it consists of gliasperin B.
10. The indicator of claim 9, wherein the plant is licorice.