Composition for reducing body fat

Abstract

To provide a composition capable of reducing body fat.SOLUTION: A composition for reducing body fat comprising corosolic acid as an active ingredient.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a composition for reducing body fat, which contains corosolic acid as an active ingredient. [Background technology]

[0002] Obesity is a condition in which adipose tissue accumulates excessively in the body, and visceral obesity, in particular the accumulation of fat in the abdominal cavity, can lead to metabolic syndrome, which is a condition in which risk factors for cardiovascular disease such as hyperglycemia, dyslipidemia, and hypertension are compounded. Therefore, preventing and improving obesity is important for maintaining and improving health.

[0003] Research into the reduction of obesity has been conducted for a long time, and for example, it is known that polyphenols derived from hops reduce body fat and / or inhibit its accumulation (Patent Document 1). In recent years, for example, a lipid-burning promoter containing arabinoxylan as an active ingredient (Patent Document 2) and an agent for improving obesity-related metabolic diseases characterized by containing guanabenz or a salt thereof as an active ingredient (Patent Document 3) have been known. However, the compositions developed so far are not necessarily sufficiently effective, and therefore, there is a need for the development of new compositions with excellent body fat-reducing effects. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2005 / 074961 [Patent Document 2] Patent Publication No. 2021-151267 [Patent Document 3] International Publication No. 2020 / 050290 Summary of the Invention [Problem to be solved by the invention]

[0005] An object of the present invention is to provide a composition capable of reducing body fat.

[0006] The present inventors have conducted extensive research to solve the above problems and have surprisingly found that corosolic acid has an excellent effect of reducing visceral fat. The present invention was completed based on this finding.

[0007] The outline of the present invention is as follows. <1> A composition for improving basal metabolism, characterized by containing corosolic acid as an active ingredient. <2> A composition for promoting PPARα gene expression, characterized by containing corosolic acid as an active ingredient. <3> A composition for promoting the beta-oxidation of fatty acids, characterized by containing corosolic acid as an active ingredient. <4> A composition for reducing body fat, characterized by containing corosolic acid as an active ingredient. <5> Characterized by visceral body fat <4> The composition for reducing body fat described above. <6> The reduction in body fat is due to one or more actions selected from the group consisting of improving basal metabolism, promoting PPARα gene expression, and promoting the beta-oxidation of fatty acids. <4> The anti-obesity composition according to claim 1. <7> The daily intake of corosolic acid is between 1.5 mg and 10 mg. <4> The composition for reducing body fat described above. <8> The daily intake of corosolic acid is between 3.5 mg and 5.5 mg. <4> ~ <7> 10. The composition for reducing body fat according to claim 1, wherein <9> Food products that contain corosolic acid as an active ingredient and are labeled as reducing visceral fat. <10> The daily intake of corosolic acid is between 3.5 mg and 5.5 mg. <9> The food product described in [Effects of the Invention]

[0008] The composition of the present invention has the effect of reducing visceral fat by improving basal metabolism, promoting PPARα gene expression, and promoting β-oxidation of fatty acids, and therefore exerts an excellent effect of reducing body fat. [Brief explanation of the drawings]

[0009] [Figure 1] FIG. 1 shows the relative expression levels of the PPARα gene in Examples and Reference Examples. [Figure 2] FIG. 2 is a diagram showing relative values of β-oxidation activity of fatty acids in Examples and Reference Examples. DETAILED DESCRIPTION OF THE INVENTION

[0010] The present invention will be described in detail below, but the present invention is not limited to the following embodiments.

[0011] The composition of the present invention is characterized by containing corosolic acid as an active ingredient. Corosolic acid is a type of triterpene and is represented by the following structural formula (1).

[0012] [ka]

[0013] The corosolic acid used in the present invention is not particularly limited as long as it is commonly used in the field of food and beverages, and examples thereof include synthetic products synthesized by methods commonly known to those skilled in the art, plants containing corosolic acid, and processed products of such plants. From the viewpoint of safety in oral ingestion, it is preferable to use plants containing corosolic acid and processed products of such plants. In the present invention, corosolic acid extracted from plants containing corosolic acid or products obtained by chemical synthesis can be used, and commercially available products may also be used.

[0014] Examples of plants containing corosolic acid used in the present invention include Chinese quince, almond, banaba leaves, cranberries, loquat leaves, perilla leaves, etc. Banaba leaves or loquat leaves are preferred because they are rich in corosolic acid, and banaba leaves are particularly preferred because of their body fat reducing effect.

[0015] Examples of processed plant products containing corosolic acid include crushed products, juices, and extracts, with extracts being preferred due to their rich corosolic acid content. The solvent used for extraction can be either polar or nonpolar, including water, methanol, ethanol, acetone, ethyl acetate, diethyl ether, n-hexane, and mixtures thereof. However, water, aqueous ethanol, or ethanol is preferred for efficient corosolic acid extraction. The extraction method is not particularly limited as long as it is generally acceptable for extracting plant components, but examples include solid-liquid extraction methods such as thermal extraction and supercritical fluid extraction. Juices and extracts can be in liquid form, but can also be used as pastes or dried powders. Pastes and dried powders can be produced using the product alone or with excipients. The resulting processed product can also be further subjected to chemical, enzymatic, and / or purification processes.

[0016] The content of corosolic acid in the composition of the present invention can be measured, for example, by LC-MS. For example, an Imtakt UK-C18 HT 3 μm (3 × 150 mm) analytical column manufactured by Intakt Corporation can be used, with 0.1% acetic acid (mobile phase A) and acetonitrile solution (mobile phase B) as the mobile phase medium, at a column temperature of 40°C and a flow rate of 0.3 mL / min. The ionization method can be electrospray, the ionization mode can be negative, and the mass number can be set to 371.5. The elution conditions can be isocratic, with mobile phase B set to 60%.

[0017] In addition to corosolic acid, other ingredients may be blended into the composition of the present invention as needed. Examples of such ingredients include excipients, lubricants, dietary fiber (e.g., soluble dietary fiber, insoluble dietary fiber), proteins, various vitamins and minerals, and microorganisms (e.g., algae and yeast). Furthermore, ingredients commonly used in the food industry, such as sweeteners, acidulants, nutritional supplements, stabilizers, binders, glossing agents, thickeners, colorants, diluents, emulsifiers, food additives, and seasonings, may also be included as needed. The content of these other ingredients may be appropriately selected depending on the form of the composition of the present invention.

[0018] As described in the Examples below, the composition of the present invention can improve basal metabolism and reduce visceral fat, thereby achieving a body fat reduction effect. Furthermore, the composition of the present invention has the effect of promoting PPARα gene expression and the effect of promoting fatty acid β-oxidation. The composition of the present invention activates PPARα and promotes fatty acid β-oxidation, thereby achieving a basal metabolism improvement effect and a visceral fat reduction effect.

[0019] The composition of the present invention has a body fat reducing effect and is therefore used as a composition for reducing body fat.

[0020] In the present invention, body fat refers to a combination of visceral fat and subcutaneous fat. Furthermore, in the present invention, "reducing body fat" also includes reducing body fat, suppressing body fat increase, anti-obesity, preventing obesity, and dieting.

[0021] The composition of the present invention is not particularly limited as long as it can be distinguished from other products in terms of its use for reducing body fat. For example, the scope of the present invention includes products that display a body fat-reducing function on the product itself, packaging, instructions, or promotional materials. Examples of such products include so-called health foods such as pharmaceuticals (including quasi-drugs), functional foods such as specified health foods, nutrient-functional foods, and functional food products whose efficacy has been recognized by a designated organization, and feed. In addition, even general foods that are manufactured and sold with the suggestion of their use for reducing body fat are included in the scope of the body fat-reducing composition of the present invention.

[0022] Specifically, compositions for reducing body fat can be advertised and promoted using phrases such as "helps reduce visceral fat," "reduces body fat (visceral fat)," "for those concerned about body fat," "for those who are slightly obese," "reduces visceral fat," "improves BMI," "reduces visceral fat (belly fat)," "reduces visceral fat by increasing basal metabolic rate," "increases basal metabolic rate," "increases energy consumption (calorie consumption)," "reduces abdominal size," and "reduces waist circumference," and such advertising and promotion can be considered to be implementations of the present invention.

[0023] The composition of the present invention is not particularly limited as long as it is in a form suitable for oral use, and examples thereof include tablets, capsules, powders, granules, liquids, granules, rods, plates, blocks, solids, pills, pastes, creams, caplets, gels, chewable tablets, and sticks. Among these, from the viewpoint of ease of administration, tablets, capsules, powders, granules, and liquids are preferred, and tablets, capsules, and granules are more preferred. Examples of oral compositions used as tablets, capsules, powders, granules, and liquids include supplements, food additives, packaged beverages filled in PET bottles, cans, bottles, etc., and powdered beverages to be dissolved in water (hot water), milk, fruit juice, etc.

[0024] The content of corosolic acid in the composition of the present invention may be appropriately determined within the range that achieves its effect. From the viewpoint of body fat reduction, the content of corosolic acid is preferably 0.00001% by mass or more, more preferably 0.0001% by mass or more, and particularly preferably 0.001% by mass or more, based on the solid content. Furthermore, from the viewpoint of ease of continuous oral intake in daily life, the content of corosolic acid is preferably 80% by mass or less, more preferably 60% by mass or less, even more preferably 50% by mass or less, and particularly preferably 40% by mass or less.

[0025] The intake amount of the composition of the present invention is not particularly limited, but from the viewpoint of body fat reduction, the intake amount of corosolic acid is preferably 0.01 mg or more per day, more preferably 1.5 mg or more, even more preferably 3 mg or more, and particularly preferably 3.5 mg or more. Furthermore, from the viewpoint of ease of continuous oral intake in daily life, the intake amount of corosolic acid is preferably 100 mg or less per day, more preferably 50 mg or less, even more preferably 10 mg or less, and particularly preferably 5.5 mg or less.

[0026] The composition of the present invention may be appropriately designed so that the daily intake of corosolic acid is the above-mentioned amount, and may be ingested in one dose or in multiple doses. That is, for example, the daily amount may be contained in one container or in 2 to 4 containers.

[0027] The packaging form of the composition of the present invention is not particularly limited and can be appropriately selected depending on the dosage form, etc., and examples include blister packs such as PTPs; strip packaging; heat seals; aluminum pouches; film packaging using plastics, synthetic resins, etc.; glass containers such as vials; and plastic containers such as ampoules. [Example]

[0028] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples, and the present invention can take various forms as long as the object of the present invention can be achieved.

[0029] <Test 1: Evaluation of the PPARα gene expression promoting effect of corosolic acid> A test was carried out to verify that the composition of the present invention has an effect of promoting PPARα gene expression.

[0030] [Test substance] Corosolic acid was used as a corosolic acid reagent (manufactured by Nagara Science). EGCG (epigallocatechin gallate) and ellagic acid were used as reference examples. EGCG was used as an EGCG reagent (manufactured by Nagara Science), and ellagic acid was used as an ellagic acid reagent (manufactured by Fujifilm Wako Pure Chemical Industries). EGCG and ellagic acid are known to promote PPARα gene expression (e.g., paragraph 0070 of JP-A-2009-507826, paragraph 0051 of JP-A-2022-031180, etc.).

[0031] [cell] Human hepatoma-derived cells (HepG2) were used as a model of human hepatocytes.

[0032] [Test method] 37℃, 5% CO by volume 2 In the incubator, 75 cm 2 HepG2 cells were cultured in a flask in 10% FBS-DMEM.

[0033] The cells were suspended by trypsinization and placed in each well of a collagen-coated 96-well plate at a concentration of 2.0 × 10 4 The cells were seeded at a density of 100 cells / well and pre-cultured for 24 hours in a 5% by volume CO 2 incubator at 37°C.

[0034] After removing the medium from each well, 100 μL of medium containing the test substance was added per well and cultured for 24 hours in a 37°C, 5% CO2 incubator. The test substance was prepared using 0.5% DMSO-10% FBS-DMEM, with a total test substance concentration of 0.1 μg / mL. A control was also prepared using 0.5% DMSO-10% FBS-DMEM without the test substance.

[0035] After 24 hours of culture, the medium was removed and the cells were washed twice with PBS. RNA was isolated using the Rneasy Mini Kit (QIAGEN), and cDNA was synthesized using ReverTra Ace qPCR RT Master Mix (TOYOBO). Quantitative real-time PCR was performed using the resulting cDNA as a template with the Rotor-Gene SYBR Green PCR Kit (QIAGEN). PPARα gene expression levels were measured using PPARα primers (Sigma). Additionally, GAPDH gene expression levels were measured as an endogenous control using GAPDH primers (QIAGEN).

[0036] The gene expression level of PPARα was evaluated by calculating the relative value when the control value was set to 1. The results are shown in Figure 1.

[0037] [Test Results] As shown in Figure 1, corosolic acid (Example 1) was found to increase the level of PPARα gene expression compared to EGCG (Reference Example 1), ellagic acid (Reference Example 2), and the control. Therefore, corosolic acid has the effect of promoting PPARα gene expression. Since the composition of the present invention exhibits an excellent effect of promoting PPARα gene expression, it can be expected to have effects of promoting fatty acid β-oxidation, improving basal metabolism, and reducing visceral fat.

[0038] <Test 2: Evaluation of the effect of corosolic acid on promoting fatty acid β-oxidation> A test was carried out to verify that the composition of the present invention has an effect of promoting the β-oxidation of fatty acids.

[0039] [Test substance] As in Test 1, a corosolic acid reagent (manufactured by Nagara Science) was used as corosolic acid. Also, as a reference example, EGCG (epigallocatechin gallate) and ellagic acid were used. An EGCG reagent (manufactured by Nagara Science) was used as EGCG, and an ellagic acid reagent (manufactured by Fujifilm Wako Pure Chemical Industries) was used as ellagic acid. EGCG and ellagic acid are known to have the effect of promoting the β-oxidation of fatty acids (e.g., paragraph 0055 of JP 2006-077026 A, paragraph 0309 of JP 2022-190124 A, etc.).

[0040] [cell] Human hepatoma-derived cells (HepG2) were used as a model of human hepatocytes.

[0041] [Test method] 37℃, 5% CO by volume 2 In the incubator, 75 cm 2 HepG2 cells were cultured in a flask in 10% FBS-DMEM.

[0042] The cells were suspended by trypsinization and placed in each well of a collagen-coated 96-well plate at a concentration of 2.0 × 10 4 The cells were seeded at a density of 100 cells / well and pre-cultured for 24 hours in a 5% CO2 incubator at 37°C. After removing the medium from each well, 150 μL / well of serum-free, phenol red-free DMEM was added and the cells were cultured for 1 hour in a 5% CO2 incubator at 37°C.

[0043] After removing the medium from each well, 100 μL / well of medium containing the test substance was added and cultured for 3 hours in a 37°C, 5% CO2 incubator. 0.5% DMSO-serum-free, phenol red-free DMEM was used as the test substance preparation medium, and the total concentration of the test substance in the medium was 1 μg / mL. Additionally, 0.5% DMSO-serum-free, phenol red-free DMEM culture medium without the test substance was added as a control.

[0044] After 3 hours of incubation, the medium was removed and the plates were washed twice with HEPES-buffered saline (HBS). 100 μL of 5 μM FAOBlue (Funakoshi Co., Ltd.) solution prepared in HBS was added to each well, and the plates were incubated at 37°C in a 5% CO2 incubator for 30 minutes.

[0045] After 30 minutes of incubation, the fluorescence intensity (Ex. 405 nm / Em. 460 nm) was measured. Based on the obtained data, the % of control (relative fluorescence intensity, with the control fluorescence intensity set at 100%) was calculated to evaluate the fatty acid β-oxidation activity. The results are shown in Figure 2. The ratio of fluorescence intensity and the ratio of fatty acid β-oxidation activity are directly proportional, so the ratio of fluorescence intensity indicates the fatty acid β-oxidation activity. % of control=(Data sample-Data blank) / (Data control-Data blank)×100

[0046] [Test Results] As shown in Figure 2, corosolic acid (Example 1) was found to increase fatty acid β-oxidation activity compared to EGCG (Reference Example 1), ellagic acid (Reference Example 2), and the control. Therefore, corosolic acid has the effect of promoting fatty acid β-oxidation. Since the composition of the present invention exhibits an excellent effect of promoting fatty acid β-oxidation, it can be expected to have effects of improving basal metabolism and reducing visceral fat.

[0047] <Test 3: Evaluation of the body fat reducing effect of corosolic acid> A clinical trial was conducted to verify that the composition of the present invention has a body fat reducing effect.

[0048] [Test substance] Corosolic acid used was that derived from a hydroethanolic extract of banaba leaves.

[0049] Examples 2 to 4 (test food groups) Corosolic acid-containing tablets were prepared. Corosolic acid-containing banaba leaf extract was mixed with maltose, cellulose, silicon dioxide, and calcium stearate, and the mixture was compressed into tablets (250 mg per tablet). The corosolic acid content per tablet was 1.0 mg in the low-dose group (Example 2), 2.9 mg in the medium-dose group (Example 3), and 4.7 mg in the high-dose group (Example 4). Comparative Example 1 (control food group) Instead of the corosolic acid-containing banaba leaf extract of Example 2, caramel color and maltitol were mixed and compressed in the same manner as in Example 2 to obtain tablets (250 mg per tablet).

[0050] [Test method] The subjects were healthy Japanese men and women aged 20 to 65, with 30 subjects per group (120 subjects in total). The final analysis included 96 subjects (23 to 26 subjects per group). The study was a randomized, double-blind, placebo-controlled, parallel-group comparative study (allocation ratio: 1:1:1:1) lasting approximately 10 weeks, consisting of a pre-observation period (10 days) and an intake period (8 weeks). During the study period, subjects were instructed to take one packet (one tablet) of the test food once a day with water or lukewarm water.

[0051] [Inspection items] Physical examination Body weight, BMI, waist circumference, hip circumference, and waist circumference / hip circumference were measured before and after 8 weeks of intake, and the results were used for statistical analysis. Abdominal visceral fat area, abdominal subcutaneous fat area, and total abdominal fat area were evaluated using CT scans (Toshiba Medical Systems Corporation: Activion 16) before and after 8 weeks of intake. The results are shown in Table 1.

[0052] [Table 1]

[0053] [Test Results] The results in Table 1 indicate that the groups that ingested banaba leaf-derived corosolic acid (Examples 2 to 4) experienced a reduction in abdominal visceral fat area and abdominal subcutaneous fat area. In particular, the group that ingested 4.7 mg / day of banaba leaf-derived corosolic acid (Example 4) showed significantly lower abdominal visceral fat area and total abdominal fat area compared to the group that ingested the control food (Comparative Example 1). It is possible that the promotion of adipose tissue metabolism during the intake period of this study led to a reduction in visceral fat accumulation prior to subcutaneous fat. The abdominal visceral fat reduction observed in this study is believed to be due to the action of corosolic acid in the banaba leaf extract. Furthermore, as shown in Experiments 1 and 2, corosolic acid promotes PPARα gene expression and fatty acid β-oxidation. Therefore, corosolic acid is thought to activate PPARα in the liver and promote fatty acid β-oxidation, thereby improving basal metabolism and reducing visceral fat.

[0054] <Production example> Based on the results of the Examples, the following Production Examples of the present invention are given below.

[0055] [Production Example 1-5: Granules] According to the formulation in Table 2, corosolic acid and other raw materials were mixed and then fluidized bed granulation was performed using a granulator to produce the granules described in Production Examples 1-5. The granules described in Production Examples 1-5 can be taken daily in an amount of 3 g dissolved in 100 ml of water, or can be taken as is without dissolving. All of the granules in Production Examples 1-5 can reduce visceral fat and are effective in reducing body fat.

[0056] [Table 2]

[0057] [Manufacturing Example 6-10: Tablets] Corosolic acid was mixed with other ingredients according to the formulation in Table 3, and then tableted using a rotary tablet press to produce tablets (250 mg) of Production Examples 6-10. One tablet of Production Examples 6-10 can be taken daily with water or the like. All tablets of Production Examples 6-10 can reduce visceral fat and are effective in reducing body fat.

[0058] [Table 3]

[0059] [Manufacturing Example 11-15: Hard Capsules] Corosolic acid was mixed with other ingredients according to the formulation in Table 4, and then coated with a film containing gelatin or hydroxypropyl cellulose to produce hard capsules (contents: 200 mg). One capsule was taken daily with water, etc. All of the hard capsules in Production Examples 11-15 were effective in reducing visceral fat and body fat.

[0060] [Table 4]

[0061] [Production example 16-20: Liquid beverage] Corosolic acid was mixed with other ingredients according to the formulation in Table 5 and bottled into a glass bottle to produce a liquid beverage (120 ml). One bottle should be consumed per day. All of the liquid beverages in Production Examples 16-20 are effective in reducing visceral fat and body fat.

[0062] [Table 5] [Industrial Applicability]

[0063] The composition of the present invention has an effect of reducing visceral fat, thereby enabling reduction of body fat and being usable as a health food or the like, and is therefore industrially useful.

Claims

1. A composition for improving basal metabolism, characterized by containing corosolic acid as an active ingredient.

2. A composition for promoting PPARα gene expression, characterized by containing corosolic acid as an active ingredient.

3. A composition for promoting the beta-oxidation of fatty acids, characterized by containing corosolic acid as an active ingredient.

4. A composition for reducing body fat, characterized by containing corosolic acid as an active ingredient.

5. 5. The composition for reducing body fat according to claim 4, wherein the body fat is visceral fat.

6. 5. The composition for reducing body fat according to claim 4, wherein the reduction in body fat is achieved by one or more actions selected from the group consisting of an action of improving basal metabolism, an action of promoting PPARα gene expression, and an action of promoting β-oxidation of fatty acids.

7. 5. The composition for reducing body fat according to claim 4, wherein the daily intake of corosolic acid is 1.5 mg or more and 10 mg or less.

8. Food products that contain corosolic acid as an active ingredient and are labeled as reducing visceral fat.

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