Anti-obesity composition and oral composition

Abstract

To provide an anti-obesity composition and an oral composition capable of accelerating a genetic expression such as Pgc1α and Ucp1 related to a brown fat cell or an energy expenditure function thereof, having an effect of accelerating the energy expenditure function in the brown fat cell.SOLUTION: The composition contains tectorigenins and pinitol. The tectorigenins is preferably tectorigenin. The composition can be suitably used as the anti-obesity composition and the oral composition.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to compositions for oral use for anti-obesity, body fat reduction, fat metabolism promotion, fat burning promotion, thermogenesis promotion, and energy consumption promotion.The present invention also relates to the use of tectorigenins and pinitol in the production of functional foods for ameliorating obesity, body fat reduction, fat metabolism promotion, fat burning promotion, thermogenesis promotion, or energy consumption promotion.The present invention also relates to methods for ameliorating obesity, body fat reduction, fat metabolism promotion, fat burning promotion, thermogenesis promotion, or energy consumption promotion using compositions containing tectorigenins and pinitol. [Background technology]

[0002] Obesity is closely related to the consumption of fat in the body, and fat consumption specifically involves the following mechanisms. When fat-mobilizing hormones such as norepinephrine are secreted, the lipolytic enzyme hormone-sensitive lipase is activated in white fat cells located subcutaneously and in the internal organs, breaking down triglycerides (neutral fats) stored in the white fat cells into glycerol and free fatty acids, which are released into the bloodstream. The free fatty acids released into the bloodstream are taken up by the mitochondria of brown fat cells located in the liver, muscles, and areas around the collarbone, chest, and shoulders, where they are consumed as heat. In this way, white fat cells store excess energy in the body as fat, while brown fat cells burn fat and generate heat. UCP1 and PGC1α are known as candidate molecules involved in the autonomous regulation of energy expenditure.

[0003] UCP uncouples oxidative phosphorylation reactions in the inner mitochondrial membrane and dissipates energy as heat. Regarding UCP1, the most representative brown fat cell, it is known that: 1. UCP1 function is reduced in obese animals; 2. UCP1 is increased in animals that do not become obese despite heavy eating; and 3. Mice with artificially reduced UCP1 gene expression become obese, while mice with high expression become lean. Furthermore, PGC1α is known to mediate UCP1 activation through stimulation of the β-adrenergic receptor in brown fat cells and to promote mitochondrial proliferation (Non-Patent Document 1). Therefore, since activating UCP1 and PGC1α is expected to have an anti-obesity effect, drugs and foods that are effective in activating UCP1 and PGC1α are being searched for.

[0004] Meanwhile, various anti-obesity agents, including that disclosed in Patent Document 1, have been known to address the issue of reducing white adipose tissue such as subcutaneous fat. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2017-171654 [Patent Document 2] Japanese Patent Application Laid-Open No. 2009-196931 [Non-patent literature]

[0006] [Non-Patent Document 1] Proceedings of the 124th Japanese Medical Association Symposium "The Science of Obesity" Japanese Medical Association, 2003, pp. 62-70 Summary of the Invention

[0007] The function of brown fat cells to consume energy from fatty acids and other substances to produce heat (hereinafter referred to as the "energy consumption function") is important for lipid metabolism. If this function can be activated, more fat in the body will be converted into heat, and it is expected that an even more effective anti-obesity effect will be achieved. However, none of the above-mentioned conventional techniques has investigated the promotion of the energy consumption function in brown adipocytes. Furthermore, the inventors have conducted research and found that conventional anti-obesity agents, which are believed to be able to reduce white adipose tissue when used alone, are not sufficient in terms of promoting the energy consumption function carried out in brown adipose tissue.

[0008] The present inventors have investigated the gene expression of proteins such as PGC1α and UCP1, which are involved in the energy consumption function in brown adipocytes, and have conducted extensive research into the composition of a composition that can promote the energy consumption function in brown adipocytes. As a result, they have surprisingly found that by combining specific ingredients, the expression of these genes can be promoted, resulting in an excellent effect of promoting the above-mentioned energy consumption.

[0009] The present invention is based on the above findings and provides an anti-obesity composition containing tectorigenins and pinitol.

[0010] The present invention provides a composition for reducing body fat or promoting fat metabolism, which contains tectorigenins and pinitol.

[0011] The present invention provides a composition for promoting fat burning, promoting thermogenesis, or promoting energy consumption, which contains tectorigenins and pinitol.

[0012] The present invention also provides an oral composition containing tectorigenins and pinitol.

[0013] The present invention also provides the use of tectorigenins and pinitol in the production of functional foods for improving obesity, reducing body fat, promoting fat metabolism, promoting fat burning, promoting thermogenesis, or promoting energy consumption.

[0014] The present invention also provides a method for improving obesity, reducing body fat, promoting fat metabolism, promoting fat burning, promoting thermogenesis, or promoting energy consumption, using a composition containing tectorigenins and pinitol. [Effects of the Invention]

[0015] According to the present invention, a useful oral composition containing a composition containing tectorigenins and pinitol can be provided. In particular, it can promote gene expression of proteins such as PGC1α and UCP1, which are involved in the energy consumption function in brown adipocytes, and have the effect of promoting energy consumption function in brown adipocytes, thereby providing effective anti-obesity compositions, body fat reduction compositions, fat metabolism promotion compositions, fat burning promotion compositions, thermogenesis promotion compositions, and energy consumption promotion compositions. Furthermore, it can provide oral compositions that promote energy consumption function in brown adipocytes through the effect of promoting the expression of the PGC1α and UCP1 genes, and are effective in anti-obesity, body fat reduction, fat metabolism promotion, fat burning promotion, thermogenesis promotion, and energy consumption promotion. Furthermore, according to the present invention, it can provide use of a composition containing tectorigenins and pinitol for obesity improvement, which can promote gene expression of PGC1α and UCP1 and promote energy consumption function in brown adipocytes, and a method for improving obesity using tectorigenins and pinitol as active ingredients. [Brief explanation of the drawings]

[0016] [Figure 1] FIG. 1 is a graph showing the expression level of the PGC1α gene in Examples and Comparative Examples. [Figure 2] FIG. 2 is a graph showing the expression level of the UCP1 gene in Examples and Comparative Examples. DETAILED DESCRIPTION OF THE INVENTION

[0017] The present invention will be described in more detail below with reference to embodiments thereof, but is not limited thereto. Hereinafter, the anti-obesity composition, body fat reduction composition, fat metabolism promotion composition, fat burning promotion composition, thermogenesis promotion composition, energy consumption promotion composition, and oral composition of the present invention will be collectively referred to as the "composition of the present invention."

[0018] (tectorigenins) Tectorigenins refer to tectorigenin, glycosides of tectorigenin, and derivatives thereof. Tectorigenin is a flavonoid found in plants such as the Iridaceae family, and has the molecular formula C 16 H 12 It is represented by the formula O6 and is sometimes called 5,7-Dihydroxy-3-(4-hydroxyphenyl)-6-methoxy-4H-1-benzopyran-4-one or 6-Methoxy-5,7-dihydroxy-3-(4-hydroxyphenyl)-4H-1-benzopyran-4-one. When simply referring to tectorigenin in this specification, this refers to the aglycone form. The chemical formula of tectorigenin is as follows:

[0019] [ka]

[0020] The term "tectorigenin glycoside" as used herein refers to the above-described tectorigenin bound to one or more sugars selected from monosaccharides and / or sugar acids. As the monosaccharide, pentose and hexose are preferred, with one or more selected from glucose, xylose, mannose, fructose, sorbose, galactose, apiose, and rhamnose being more preferred, and one or two selected from glucose and xylose being particularly preferred. Furthermore, as the sugar acid, uronic acid is preferred, with glucuronic acid and galacturonic acid being particularly preferred. In glycosides, these sugars and / or sugar acids are usually bound to the hydroxyl groups at the 4'- and / or 7'-positions of tectorigenin. In the present invention, those bound to the hydroxyl group at the 7'-position are preferred. The number of sugars and / or sugar acids in a tectorigenin glycoside (also referred to as the number of sugar and / or sugar acid bonds) can be, for example, from 1 to 5, preferably from 1 to 4, and particularly preferably from 1 to 3. The number of sugars and / or sugar acids refers to the total number of sugars and / or sugar acids. Here, "n sugars bound to tectorigenin" may mean that n sugar linkers are bound, or the total number of sugars bound to different sites on tectorigenin may be n. The same applies when the glycoside contains a sugar acid.

[0021] Derivatives of tectorigenin or its glycosides include, for example, compounds in which one or more hydrogen atoms in tectorigenin or its glycosides are substituted with substituents. Examples of such substituents include amino groups, sulfonic acid groups, alkyl groups having 1 to 6 carbon atoms, and alkoxy groups having 1 to 4 carbon atoms. The number of the above-mentioned substituents in tectorigenins is preferably 8 or less, more preferably 4 or less, and most preferably 0.

[0022] The composition of the present invention may use only one of the above-mentioned tectorigenin, its glycoside, and derivatives thereof as the tectorigenin, or a mixture of two or more of them. In particular, in terms of anti-obesity effects, it is preferable that the tectorigenin in the composition of the present invention is tectorigenin and / or its glycoside. In particular, it is preferable that the composition of the present invention contains tectorigenin as the tectorigenin, in terms of its excellent effect of promoting energy consumption in brown adipocytes and ease of availability. In the composition of the present invention, the tectorigenin may be an organic synthetic product or may be extracted from a plant or the like.

[0023] (Pinitol) Pinitol, also known as 3-O-methyl-D-chiro-inositol, is a type of inositol. Pinitol is found in plants such as carob and clover. In the composition of the present invention, pinitol may be an organically synthesized product or may be extracted from a plant or the like.

[0024] In the present invention, a specific range of the content ratio of tectorigenins to pinitol is preferred because it promotes excellent gene expression of PGC1α and UCP1 and further enhances the energy consumption activity in brown adipocytes. To enhance the energy consumption activity in brown adipocytes, the content ratio of tectorigenins to pinitol is preferably 1:0.01 to 50 (tectonic:pinitol) by mass, more preferably 1:0.05 to 30 (tectonic:pinitol), and particularly preferably 1:0.1 to 26 (tectonic:pinitol). For the same reason, when tectorigenin is contained as the tectorigenin, the content ratio of tectorigenin to pinitol is preferably 1:0.01 to 50 (tectonic:pinitol) by mass, and is particularly preferably 1:0.5 to 30 (tectonic:pinitol) by mass, particularly because it promotes the energy consumption activity in brown adipocytes by promoting UCP1 gene expression.

[0025] The amount of tectorigenins in the composition of the present invention can be measured by high performance liquid chromatography (HPLC). For example, a YMC-Pack ODS AM12S05-2546WT (φ4.6 × 250 mm) manufactured by YMC Co., Ltd. may be used, and the mobile phase may be a mixture of acetonitrile / water / acetic acid (mobile phase A volume ratio = 15:85:0.1, mobile phase B volume ratio = 35:65:0.1), with a column temperature of 35°C and a flow rate of 1.0 ml / min. The gradient conditions can be as shown in Table A below.

[0026] [Table A]

[0027] The amount of pinitol in the composition of the present invention can be measured by HPLC. For example, a column such as InertSustain NH2 (3 μm, φ3 × 250 mm) manufactured by GL Sciences Inc. can be used, the mobile phase can be 1 mM hydrochloric acid / chromatographic acetonitrile (volume ratio = 30 / 70), the column temperature can be 40°C, and the flow rate can be 0.5 ml / min.

[0028] The composition of the present invention may contain, in addition to tectorigenins and pinitol, other commonly used ingredients within a range that does not impair the effects of the present invention. Such ingredients include various excipients, binders, glossing agents, lubricants, stabilizers, diluents, bulking agents, thickeners, emulsifiers, antioxidants, pH adjusters, colorants, flavorings, additives, etc. The content of other ingredients can be appropriately selected depending on the form of the composition of the present invention, etc.

[0029] The composition of the present invention can be used for both oral and parenteral administration. Examples of parenteral preparations include those administered by direct administration to the stomach using a catheter or the like. However, oral administration of the composition of the present invention is preferred because it allows the effect of the present invention on promoting energy consumption to be easily achieved. When used as an oral preparation, examples of the form include tablets, capsules, powders, granules, liquids, granules, rods, plates, blocks, solids, pills, pastes, creams, caplets, gels, chewable tablets, and sticks. Among these, tablets, capsules, powders, granules, and liquids are particularly 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, and the like; and powdered beverages to be dissolved in water (hot water), milk, fruit juice, and the like. These are preferred because they are easy to consume during meals and can enhance palatability.

[0030] Specific examples of the oral composition of the present invention include foods as edible compositions. Examples of foods include tea bags, breads, confectioneries, noodles, and other foods, as well as cooked foods. Examples of breads and confectioneries include breads such as white bread, sweet buns, French bread, English bread, muffins, steamed buns, donuts, and waffles; cakes such as butter cake, sponge cake, chiffon cake, and pancakes; frozen desserts such as chocolate, sorbet, and ice cream; jellies; and cookies. Examples of noodles include udon and somen noodles. Examples of cooked foods include soups such as curry, stew, miso soup, and vegetable soup, as well as their bases and seasonings.

[0031] The composition of the present invention preferably contains pinitol and tectorigenins in a total amount of 0.001% by mass to 40% by mass in terms of promoting energy consumption in brown adipocytes and facilitating continuous oral intake in daily life, more preferably 0.005% by mass to 35% by mass, and particularly preferably 0.01% by mass to 30% by mass. The solid content is the amount of the composition excluding water.

[0032] When the composition of the present invention is orally ingested, the oral dosage is preferably from about 0.1 mg to 500 mg, more preferably from 0.5 mg to 300 mg, of the total amount of pinitol and tectorigenins per adult per day, in terms of promoting energy consumption in brown adipocytes and ease of continuous oral intake in daily life. Furthermore, the single intake amount of the composition of the present invention is preferably from about 1 mg to 300 mg, of the total amount of pinitol and tectorigenins per adult per day.

[0033] Specific examples of the use of the composition of the present invention include so-called health foods such as pharmaceuticals (including quasi-drugs), general foods, foods with nutrient function claims, foods for specified health uses whose efficacy has been approved by a designated organization, and foods with functional claims. Foods that display efficacy are sometimes collectively referred to as "functional foods."

[0034] As shown in the Examples below, the composition of the present invention can promote energy consumption in brown adipocytes and brown adipose tissue containing them by promoting the expression of genes encoding multiple proteins related to energy consumption in brown adipocytes, such as the PGC1α gene and the UCP1 gene.

[0035] PGC1α (PPARγ coactivator 1α), a protein encoded by the PGC1α gene, was identified as a transcriptional coactivator that activates transcription by the nuclear receptor PPARγ. PGC1α activates the transcription factors NRF (nuclear respiratory factor) 1 / NRF2, which in turn activates mitochondrial biogenesis by promoting the transcription of TFAM (mitochondria transcription factor). It is known that promoting PGC1α gene expression increases mitochondria. Furthermore, PGC1α is a factor that binds nuclear receptors in brown adipocytes and is involved in promoting UCP1 gene expression. Therefore, promoting PGC1α gene expression may increase mitochondria in brown adipocytes and promote energy consumption through promoting UCP1 gene expression. The PGC1α gene is sometimes referred to as Pgc1α.

[0036] The UCP1 gene encodes the molecule uncoupling protein 1 (UCP1) and is sometimes referred to as Ucp1. In brown adipocytes, the UCP1 gene is expressed specifically in the mitochondria of those cells. The UCP1 gene has the activity of uncoupling oxidative phosphorylation in mitochondria, and when activated, the energy generated by the oxidative degradation of fatty acids and glucose is directly converted into heat and dissipated without being used for ATP synthesis. Promoting the expression of the UCP1 gene can promote fatty acid consumption in brown adipocytes, thereby suppressing or eliminating obesity.

[0037] The effect of the present invention may be to promote the expression of only one of the PGC1α gene and the UCP1 gene, or may be to promote the expression of both the PGC1α gene and the UCP1 gene. The compositions of the present invention can be used for various purposes, such as promoting fat burning, promoting thermogenesis, promoting fat metabolism, reducing body fat, and promoting energy consumption, in addition to promoting the expression of the PGC1α gene and the UCP1 gene. Fat burning promotion refers to promoting the conversion of fatty acids into heat by brown adipocytes. Promoting thermogenesis refers, for example, to promoting thermogenesis from neutral fat by brown adipocytes. Fat metabolism promotion refers, for example, to the qualitative conversion of obese adipocytes or adipose tissue into small, normal adipocytes, or the metabolic activity of fatty acids by brown adipocytes. Body fat reduction refers, for example, to the reduction of body fat by converting fatty acids into heat. Anti-obesity applications refer, for example, to the reduction of body fat or the suppression of body fat gain by promoting the metabolism of fatty acids in brown adipocytes or brown adipose tissue. The promotion of energy consumption refers to, for example, promoting the consumption of chemical energy from fatty acids by brown adipocytes. These actions are mediated by the activation of PGC1α and UCP1 in brown adipocytes. In the present invention, brown fat cells are located in the liver, muscles, and also in the vicinity of the collarbone, chest (around the chest), shoulder (scapula and surrounding areas), and the like.

[0038] Furthermore, the composition of the present invention may be any product that can be distinguished from other products in that it is used for promoting lipolysis, anti-obesity, and dieting. For example, an indication that it has the function of promoting lipolysis means an indication that appeals to subjects who are concerned about body fat, such as those who are concerned about obesity, those who are concerned about their stomach area, those who are concerned about their weight, and those who are concerned about abdominal fat (visceral fat, subcutaneous fat, etc.), or an indication that it is useful for reducing body fat, such as helping to lose weight, helping to reduce abdominal fat (visceral fat, subcutaneous fat, etc.), helping to reduce waist circumference, supporting obesity reduction, supporting dieting, making it easier to consume fat, making it easier to burn fat, helping to consume fat, helping to burn fat, promoting fat breakdown, making it easier to break down fat, helping to break down fat, etc. [Example]

[0039] The present invention will be described in more detail below with reference to examples. However, the scope of the present invention is not limited to such examples. Hereinafter, unless otherwise specified, "%" means % by mass and "parts" means parts by mass.

[0040] Measurement of PGC1α gene expression level Example 1 and Comparative Examples 1 and 2 (1) As test substances, tectorigenin (manufactured by Tokyo Chemical Industry Co., Ltd.) was used as a tectorigenin, and D-pinitol (manufactured by Tokyo Chemical Industry Co., Ltd.) was used as pinitol. (2) The proliferation medium was a proliferation medium, the differentiation induction medium was a differentiation induction medium, and the adipocyte maintenance medium was a differentiation maintenance medium. All three media were from the Brown Adipocyte Culture Kit F-8 (rat) (Cosmo Bio, derived from interscapular brown adipose tissue). Brown adipocytes derived from interscapular brown adipose tissue are frequently used to investigate brown adipocyte function related to fat burning.

[0041] (3) Differentiation-inducing medium containing test substances was prepared as follows: Each test substance was dissolved in DMSO, then diluted with differentiation-inducing medium to a final DMSO concentration of 0.5 vol%, and filter-sterilized. After sterilization, tectorigenin and pinitol were diluted with differentiation-inducing medium containing 0.5 vol% DMSO to the concentrations shown in Table 1 below.

[0042] (4) Differentiation maintenance medium containing the test substance was prepared as follows: Each test substance was dissolved in DMSO, then diluted with the differentiation maintenance medium to a final DMSO concentration of 0.5 vol%, and filter-sterilized. After sterilization, tectorigenin and pinitol were diluted with the differentiation maintenance medium containing 0.5 vol% DMSO to the concentrations shown in Table 1 below.

[0043] (5) Rat brown adipose tissue precursor cells (Cosmo Bio) were cultured in a collagen-coated 24-well plate at 3.0 × 10 cells per well using growth medium. 4 The cells were seeded at 500 μl / well so that the cells were 100 cells / well, and cultured in a 37°C, 5 (vol / vol)% CO 2 incubator until the cell density reached approximately 90%.

[0044] (6) After removing the medium, 500 μL / well of differentiation-inducing medium containing the test substance was added, and the cells were cultured in a 37°C, 5 (vol / vol)% CO2 incubator for 48 hours to induce differentiation into brown adipocytes.

[0045] (7) After removing the medium, 500 μL / well of differentiation maintenance medium containing the test substance was added, and the cells were cultured in a 37°C, 5 (vol / vol)% CO 2 incubator for 72 hours.

[0046] (8) After removing the medium, 500 μL / well of differentiation maintenance medium containing the test substance was added, and the cells were further cultured for 72 hours in a 37°C, 5 (vol / vol)% CO 2 incubator.

[0047] (9) After removing the medium, the cells were washed twice with PBS, and RNA was recovered using an RNeasy Mini Kit (QIAGEN). cDNA was synthesized using ReverT raAce® qPCR RT Master Mix (TOYOBO).

[0048] (10) Using the obtained cDNA as a template and primers for the PGC1α gene (QIAGEN), quantitative real-time PCR was performed with a QuantiNova SYBR Green PCR Kit (QIAGEN) to measure the expression level of PGC1α mRNA. The relative values ​​of the expression levels of PGC1α mRNA in Example 1 and Comparative Examples 1 and 2, with the value for Comparative Example 1 set at 1, are shown in Figure 1.

[0049] [Table 1]

[0050] Measurement of UCP1 gene expression level [Examples 2 to 4 and Comparative Examples 3 and 4] Primers for the UCP1 gene (manufactured by QIAGEN) were used instead of Pgc1α. Test substances with the compositions shown in Table 2 below were used. The concentrations of the test substances in the differentiation-inducing medium and differentiation-maintaining medium were 50 μM. Except for this, the expression levels of UCP1 mRNA were measured in the same manner as in Example 1 and Comparative Examples 1 and 2. The expression levels of UCP1 mRNA in Examples 2 to 4 and Comparative Examples 3 and 4 are shown in Figure 2 as relative values, with the value for Comparative Example 3 set to 1.

[0051] [Table 2]

[0052] As shown in Figures 1 and 2, when brown adipocytes were induced to differentiate and maintained in the presence of the composition of the present invention, which combined tectorigenins and pinitol, the expression levels of the PGC1α gene and the UCP1 gene were significantly higher than when tectorigenins or pinitol were used alone. In particular, the UCP1 gene, which plays an important role in energy consumption, showed a synergistic expression-promoting effect when tectorigenins and pinitol were used in combination. Therefore, it is clear that the composition of the present invention exhibits an excellent effect in promoting energy consumption in brown adipocytes and is an excellent anti-obesity agent.

[0053] Granules (3000 mg per packet) containing the following ingredients were prepared. By suspending one packet of the resulting granules in water twice a day and taking it, an excellent anti-obesity effect can be obtained.

[0054] [Table 3]

[0055] Granules (2000 mg per packet) containing the following ingredients were manufactured. By suspending one packet of the resulting granules in hot water and taking it three times a day, an excellent anti-obesity effect can be obtained.

[0056] [Table 4]

[0057] Tablets (200 mg each) containing the following ingredients were prepared. Taking four tablets twice a day provides excellent anti-obesity effects.

[0058] [Table 5]

[0059] Tablets (300 mg each) containing the following ingredients were manufactured. Taking five of the resulting tablets once a day provides excellent anti-obesity effects.

[0060] [Table 6]

[0061] Capsules (300 mg each) containing the following ingredients were prepared. Taking two capsules twice a day provides excellent anti-obesity effects.

[0062] [Table 7] [Industrial Applicability]

[0063] The composition of the present invention is expected to have a high level of anti-obesity effect and is therefore highly useful industrially.

Claims

[Claim 1] Contains tectorigenin and pinitol, An oral composition having a mass ratio of tectorigenin to pinitol of 1:1 or more and 2:26 or less.

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