Peroxisome proliferation agent-responsive receptor activation composition

Abstract

To provide a peroxisome proliferating agent-responsive receptor activation composition that is suitable for adding to food or the like in order to ingest, and is also suitable for continuous ingestion due to its excellent safety.SOLUTION: Provided is a peroxisome proliferating agent-responsive receptor activation composition that contains sucrose fatty acid ester as an active ingredient.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition for activating peroxisome proliferator-activated receptors (PPARs), which exhibits the property of promoting the activation of PPARs in vivo. [Background technology]

[0002] Soybeans have been eaten as a traditional ingredient in the Japanese diet since ancient times. Recently, it has become clear that soybeans contain many functional ingredients that contribute to maintaining and promoting health, and their use is being reconsidered. The health benefits of soybeans are well known for their lipid metabolism improving effect. For example, it has been shown that soybean isoflavones lower blood lipid levels, soybean dietary fiber (okara) lowers serum total cholesterol levels, soybean protein lowers blood cholesterol and neutral fat levels, inhibits body fat accumulation, and linoleic acid and alpha-linolenic acid lower serum cholesterol levels.

[0003] Recent studies have also revealed that activation of the nuclear receptor PPAR contributes to the control of lipid metabolism. Therefore, active exploration and development of PPAR activators that activate lipid metabolism has been conducted. Examples of such activators include gallic acid esters (Patent Document 1), galloyl tannins, catechin and epicatechin (Patent Document 2), organic solvent extracts of soybean fermented by tempeh bacteria (Patent Document 3), and monoacylglycerols whose constituent fatty acids are oleic acid or linoleic acid (Patent Document 4).

[0004] In addition, synthetic PPAR activators such as fibrate compounds, thiazolidine derivatives, fatty acids, leukotriene B4, indomethacin, ibuprofen, fenoprofen, and 15-deoxy-Δ12,14-prostaglandin J2 are known to activate PPAR. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2002-80362 A [Patent Document 2] JP 2011-105763 A [Patent Document 3] JP 2012-171911 A [Patent Document 4] JP 2001-354558 A Summary of the Invention [Problem to be solved by the invention]

[0006] Under these circumstances, the inventors have conducted extensive research into a composition for activating PPAR that is suitable for addition to foods and other foods without affecting taste or flavor, and that is also safe and suitable for continuous intake.

[0007] An object of the present invention is to provide a composition for activating PPAR, which is suitable for ingestion by addition to food etc. and further suitable for continuous ingestion due to its excellent safety.

[0008] In addition, the present invention focuses on soybeans, which are excellent in lipid metabolism, and provides a composition for activating PPAR, which is made from soybeans and has an improved PPAR activating effect. [Means for solving the problem]

[0009] The present inventors have conducted extensive research to solve the above problems. In the course of their research, they have unexpectedly discovered that sucrose fatty acid esters have a property of promoting PPAR activation, which was previously unknown. Sucrose fatty acid esters are suitable for ingestion by adding them to foods and the like because they do not affect the taste or flavor, and are also excellent in safety and suitable for continuous ingestion. Therefore, by containing sucrose fatty acid esters as an active ingredient, a composition for activating PPAR that is superior to conventional compositions can be obtained. Furthermore, it was also found that the PPAR activity of processed foods made from soybeans could be significantly improved by adding sucrose fatty acid esters to the soybean raw material.

[0010] That is, the gist of the present invention is the following [1] to [3]. [1] A composition for activating PPAR, comprising a sucrose fatty acid ester as an active ingredient. [2] The composition for activating PPAR described in [1], wherein the constituent fatty acid of the sucrose fatty acid ester is stearic acid. [3] A composition for activating PPAR according to [1] or [2], which is a mixture of the sucrose fatty acid ester and soybean flour. Effect of the Invention

[0011] Since the composition for activating PPAR of the present invention contains sucrose fatty acid ester as an active ingredient, it does not affect the taste or flavor and is suitable for being added to food and ingested, and furthermore, since it is highly safe, it is suitable for continuous ingestion. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Next, an embodiment of the present invention will be described, although the present invention is not limited to this embodiment.

[0013] As described above, the composition for activating PPAR of the present invention contains a sucrose fatty acid ester as an active ingredient. Sucrose fatty acid esters can be obtained by reacting the hydroxyl groups of sucrose (sucrose) with the carboxyl groups of fatty acids to form ester bonds. There are eight hydroxyl groups in one sucrose molecule, and since there are various types of fatty acids, various sucrose fatty acid esters can be synthesized depending on the number of ester bonds and the type of fatty acid. Examples of the constituent fatty acids of the sucrose fatty acid esters include saturated and unsaturated fatty acids having 12 to 22 carbon atoms, such as lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, behenic acid, and erucic acid. These can be used alone or in combination of two or more. Among them, it is preferable to use stearic acid, which is a saturated fatty acid having 18 carbon atoms.

[0014] In the composition for activating PPAR of the present invention, the above-mentioned sucrose fatty acid esters are used alone or in combination of two or more kinds. In particular, by using two or more different sucrose fatty acid esters in combination, a composition for activating PPAR that shows performance more suitable for individual needs can be obtained.

[0015] When the composition for activating PPAR of the present invention is in liquid form, the content of sucrose fatty acid ester is preferably in the range of 0.01 to 20 mg / ml, more preferably 0.01 to 10 mg / ml, from the viewpoint of the effectiveness of PPAR activity. The composition for activating PPAR of the present invention may be in any form, such as solid, powder, or liquid, as long as it can be orally ingested.

[0016] In addition, when the composition for activating PPAR of the present invention is a mixture of sucrose fatty acid ester and soybean flour, it is preferable because it additively or synergistically enhances the PPAR activating effect of isoflavones and saponins contained in the soybean flour. In the present invention, soybean flour refers to powdered soybeans from which the seed coat has been removed.

[0017] The PPAR activating composition of the present invention obtained in this manner can be taken orally continuously to promote the activation of PPAR, thereby exerting excellent effects in the prevention and improvement of hyperlipidemia, inflammatory diseases (including skin inflammation), insulin resistance diseases (diabetes, arteriosclerosis, etc.), stroke, biological rhythm disorders (biorhythm disorder diseases), obesity, promotion of body fat burning, reduction of blood triglycerides, suppression of fatty liver, improvement of endurance, differentiation of epidermal cells and differentiation of melanocytes (skin whitening effect by inducing differentiation of epidermal cells and inhibiting differentiation of melanocytes), promotion of keratinization, development and repair of the epidermis, etc.

[0018] In addition, the PPAR activating composition of the present invention can be directly orally ingested as a mixture of sucrose fatty acid ester and soybean flour as described above, or an extract of the active ingredient can be dissolved or dispersed in a suitable liquid carrier or mixed with a suitable powder carrier and then orally ingested.

[0019] Examples of pharmacologically acceptable carriers include excipients, lubricants, binders and disintegrants in solid preparations, and solvents, solubilizing agents, suspending agents, tonicity agents, buffers and soothing agents in liquid preparations.

[0020] In addition, when the composition for activating PPAR of the present invention is formulated, various ingredients that are usually used in formulations are optionally used, such as starch, dextrin, lactose, corn starch, inorganic salts, etc. These may be used alone or in combination of two or more kinds.

[0021] The dosage form of the above-mentioned preparations may be, for example, ampoules, tablets, capsules, granules, fine granules, powders, drinks, and the like.

[0022] Furthermore, the composition for activating PPAR of the present invention can also be provided in a form incorporated into food and drink products. Examples of the above-mentioned food and drink products include health foods (tablets, powders, granules, concentrated liquids), soft drinks, foods for specified health uses, lactic acid bacteria-fermented foods (such as yogurt), drinks, tea, milk, pudding, jelly, candy, gum, chocolate, soup, cookies, snack foods, wine, shochu, sake, dressings, boiled beans, tofu, natto, soy milk, roasted beans, dried beans, miso, and the like. By continuously consuming these food and drink products in the same manner as ordinary food and drink products, the activation of PPAR can be promoted, and various effects as described above (prevention and improvement of hyperlipidemia, obesity, development and repair of the epidermis, etc.) can be obtained.

[0023] The composition for activating PPAR of the present invention is gentle to the human body without causing side effects and can exhibit PPAR activation effects and preventive and improvement effects on various diseases related thereto (particularly, obesity and hyperlipidemia, etc.). In addition, the above effects can be obtained not only in humans but also in animals such as pets and livestock, and the dosage thereof is appropriately set according to conditions such as the difference in the organism to be administered, the gender, weight, age, etc. of the person to be administered. And as described above, since the composition for activating PPAR of the present invention can also obtain an obesity inhibitory effect, etc. in animals such as pets and livestock, it can also be provided in a form incorporated into pet food and feed.

Examples

[0024] Next, the examples will be described together with the comparative examples. However, the present invention is not limited to these examples. Also, in the following description, “%” means by weight unless otherwise specified.

[0025] <PPAR Activation Test> By the following method, an activation test of nuclear receptors (PPARα, PPARδ) was performed using a reporter gene assay. The cell line used was CV-1 (obtained by sublicense from the National Institute of Biomedical Innovation, Health and Nutrition). The above cell line was adjusted to 2.0×10 5The cells were seeded onto a 6-well plate at 1 cell / well and cultured for one day in Dulbecco's modified Eagle's medium (containing 10% fetal bovine serum (FBS) and 2 mol / l L-glutamine). Next, a chimeric protein expression plasmid (pGal4DBD / NR(LBD)) of the DNA binding domain of the Gal4 gene (Gal4-DBD) and the ligand binding domain (NR-LBD) of various human nuclear receptors (PPARα, PPARδ), a reporter plasmid (pGal4-Luc) containing the Gal4 gene DNA response element and the firefly luciferase gene, and an internal standard plasmid (pGL4.75hRluc-CMV; Promega) in which a gene constitutive expression promoter (CMV) was linked upstream of the Renilla luciferase gene as an internal standard were mixed in a weight ratio of 1:0.9:0.1 and dissolved in Opti-MEM medium (GIBCO), to which the gene introduction reagent X-tremeGENE HP (Roche) was added, and the mixture was added to the cultured cells and cultured to introduce the gene. Next, the transfected cells were detached with trypsin, washed, and then placed in a 96-well plate at 1.6 × 10 4 The cells were seeded again at 100 cells / well. At this time, the culture medium was replaced with Dulbecco's modified Eagle medium (phenol red-free, 10% activated charcoal-treated FBS, SIGMA) containing the test sample, and the cells were cultured for another 48 hours. The cells were washed with phosphate buffered saline (PBS) and lysed using a dual luciferase assay system (Promega), after which a substrate solution containing luciferin was added and firefly luciferase activity and Renilla luciferase activity were measured using a plate reader (Ferrios AB-2350, ATTO). Note that the PPAR activation test was performed after confirming that no significant damage was observed in the cells (no significant decrease in Renilla luciferase activity value was observed). The above procedure was carried out using three wells per sample (including the negative control), and the average value of the three wells was used as data. The transcriptional activity (luciferase activity) of the nuclear receptor-dependent gene was defined as shown in the following formula (1).

[0026] Nuclear receptor-dependent transcriptional activity value (internally standardized value) = (Firefly luciferase activity by Gal4-Luc) / (Renilla luciferase activity by hRluc-CMV)……(1)

[0027] [Example 1] <Measurement of PPAR activity of sucrose fatty acid ester> An ethanol extract of sucrose stearate was obtained, and this was used as a test sample. The PPAR activity of the test sample was measured by the method described in the <PPAR activation test>. The test sample was prepared as follows. That is, first, two types of sucrose stearates (Ryoto Sugar Ester S-370, Ryoto Sugar Ester S-770, manufactured by Mitsubishi Chemical Foods Co., Ltd.) were mixed at a weight ratio of 1:1, 10 times the amount of 70% ethanol solution was added, and after stirring at room temperature (23°C) for 3 hours, it was filtered through a 0.2 μm membrane filter and freeze-dried. What was dissolved by adding dimethyl sulfoxide (DMSO) to make this 80 mg / ml was used as the test sample. Each test sample was adjusted to a final concentration of 0.04 mg / ml, 0.16 mg / ml, and 0.40 mg / ml, and a 0.5% dimethyl sulfoxide (DMSO) solution was used as a negative control (NC). Note that the evaluation of PPARα activity and PPARδ activity was expressed as a ratio to the negative control (activity value in the sample / activity value in the negative control), and when this numerical value was 1.2 or more, it was defined as significant activity. The results are shown in Table 1 below.

[0028]

Table 1

[0029] From the results in Table 1, it is seen that the activity of PPARα and PPARδ is improved in proportion to the concentration of sucrose stearate. In addition, even at a low concentration of sucrose stearate (0.04 mg / ml), the activity of PPARα and PPARδ is improved.

[0030] [Example 2] <Measurement of PPAR activity in soy flour containing sucrose fatty acid esters> An ethanol extract of soybean flour containing sucrose stearate was obtained and used as a test sample to measure the PPAR activity of the test sample in accordance with the method described in Example 1. The PPARα activity and PPARδ activity were evaluated in the same manner as in Example 1. The test sample was prepared as follows. First, two types of sucrose stearates (Ryoto Sugar Ester S-370 and Ryoto Sugar Ester S-770, manufactured by Mitsubishi Chemical Foods Corporation) were mixed in a weight ratio of 1:1, and soy flour was further mixed, stirred and dissolved in hot water, and homogenized with a homogenizer to prepare a soy liquid. This soy liquid was freeze-dried, and a 10-fold amount of 70% ethanol solution was added to the freeze-dried product obtained, and the mixture was stirred at room temperature for 3 hours, centrifuged, and the supernatant after centrifugation was filtered through a 0.2 μm membrane filter and freeze-dried. This was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 400 mg / ml and used as a test sample. The final concentrations of the test samples were adjusted to 0.04 mg / ml for sucrose stearate and 2.0 mg / ml for soybean flour, and the negative control (NC) was the same as in Example 1. The results are shown in Table 2 below.

[0031] [Comparative Example 1] <Measurement of PPAR activity in soybean flour> An ethanol extract of soybean flour was obtained and used as a test sample, and the PPAR activity of the test sample was measured in accordance with the method described in Example 1. The PPARα activity and PPARδ activity were evaluated in the same manner as in Example 1. The test samples were obtained in the same manner as in Example 2, except that sucrose stearate was not used. The final concentration of the test sample was adjusted to 2.0 mg / ml of soybean flour, and the negative control (NC) was the same as in Example 1. The results are shown in Table 2 below.

[0032] [Table 2]

[0033] The results in Table 2 above show that the activity of PPARα and PPARδ in Example 2, in which soybean flour was used in combination with sucrose stearate, is significantly improved compared to the activity of PPARα and PPARδ when the concentration of sucrose stearate is 0.04 mg / ml, as shown in Table 1. In particular, Example 2 shows a high synergistic effect in the activity of PPARα. In addition, comparative example 1, in which only soy flour was used without using sucrose stearate, showed relatively high PPARα activity and PPARδ activity, but example 2 shows that even a small amount of sucrose stearate (concentration of sucrose stearate: 0.04 mg / ml) increases the level of PPARα and PPARδ activity when used in combination with soy flour.

[0034] The effect of improving PPAR activity seen in the above Example 1 and Example 2 can be considered to be an effect of improving PPAR activity when the composition for activating PPAR of the present invention is orally ingested as a food, which is one embodiment of the composition for activating PPAR of the present invention. In addition, since the active ingredient of the composition, sucrose fatty acid ester, does not affect the taste or flavor, it is considered to be suitable for being added to food and ingested. Furthermore, from the results of the above experiments, it can be expected that the composition for activating PPAR of the present invention, which is used in combination with soybean flour as in Example 2, will have a higher effect of improving PPAR activity when orally ingested as a food. [Industrial Applicability]

[0035] The composition for activating PPAR of the present invention is suitable for ingestion by adding to food, etc., since the active ingredient, sucrose fatty acid ester, does not affect the taste or flavor, and is also suitable for continuous ingestion due to its excellent safety. Therefore, the composition for activating PPAR of the present invention can be applied as food and drink, and can also be powdered, tableted, encapsulated, etc., and used as supplements, medicines, food additives, etc., as needed.

Claims

1. A composition for activating a peroxisome proliferator-activated receptor, comprising a sucrose fatty acid ester as an active ingredient.

2. 2. The composition for activating a peroxisome proliferator-activated receptor according to claim 1, wherein the constituent fatty acid of the sucrose fatty acid ester is stearic acid.

3. 3. The composition for activating a peroxisome proliferator-activated receptor according to claim 1, which is a mixture of said sucrose fatty acid ester and soybean flour.

Images