Composition for activating peroxisome proliferator-responsive receptors

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 receptor (PPAR), which exhibits the property of promoting the activation action of PPAR in vivo.

Background Art

[0002] Soybeans have been consumed as a food ingredient in traditional Japanese cuisine since ancient times. Recently, it has been revealed that they contain many functional components that contribute to the maintenance and improvement of health, and their utilization is being reevaluated. Regarding the health functions of soybeans, the lipid metabolism-improving effect is well known. For example, the effect of lowering blood lipid concentration by soy isoflavones, the effect of lowering serum total cholesterol by soy dietary fiber (okara), the effect of lowering the concentrations of blood cholesterol and blood triglycerides by soy protein, the effect of suppressing body fat accumulation, and the effect of lowering serum cholesterol concentration by linoleic acid and α-linolenic acid have been revealed.

[0003] In addition, recent studies have revealed that the activation of the nuclear receptor PPAR contributes to the regulation of these lipid metabolisms. Therefore, the search and development of PPAR activator substances that activate lipid metabolism have been actively carried out. As an example, gallic acid esters (Patent Document 1), galloyl tannins, catechins and epicatechins (Patent Document 2), organic solvent extracts of soybean tempeh fermented products (Patent Document 3), monoacylglycerols whose constituent fatty acids are oleic acid or linoleic acid (Patent Document 4), etc. have been reported.

[0004] In addition, conventionally, for substances that activate PPAR, synthetic PPAR activators such as fibrate compounds, thiazolidine derivatives, fatty acids, leukotriene B4, indomethacin, ibuprofen, fenoprofen, 15-deoxy-Δ12,14-prostaglandin J2 are known.

Prior Art Documents

Patent Documents

[0005] [Patent Document 1] Japanese Patent Publication No. 2002-80362 [Patent Document 2] Japanese Patent Publication No. 2011-105763 [Patent Document 3] Japanese Patent Publication No. 2012-171911 [Patent Document 4] Japanese Patent Publication No. 2001-354558 [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] Under these circumstances, the inventors have conducted extensive research on PPAR activating compositions that are suitable for ingestion by adding them to food products without affecting taste or flavor, and that are also highly safe and suitable for continuous intake.

[0007] The present invention aims to provide a PPAR activating composition that is suitable for ingestion by adding it to food, and is also 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 PPAR activating composition made from soybeans with further improved PPAR activating activity. [Means for solving the problem]

[0009] The inventors diligently conducted research to solve the aforementioned problems. In the course of this research, they unexpectedly discovered that sucrose fatty acid esters have the property of promoting the activation of PPARs, which was previously unknown. Sucrose fatty acid esters are suitable for ingestion by adding them to food products, etc., as they do not affect the taste or flavor, and furthermore, they are highly safe and suitable for continuous intake. Therefore, by including sucrose fatty acid esters as an active ingredient, an unprecedented and superior PPAR activating composition can be obtained. Furthermore, we discovered that adding sucrose fatty acid ester to soybean raw materials can significantly improve the PPAR activity of processed foods made from soybeans.

[0010] In other words, the gist of the present invention is as follows: [1] to [3]. [1] A PPAR activating composition characterized by containing a sucrose fatty acid ester as an active ingredient. [2] The PPAR activating composition according to [1], wherein the constituent fatty acid of the sucrose fatty acid ester is stearic acid. [3] A PPAR activating composition according to [1] or [2], which is a mixture of the sucrose fatty acid ester and soy flour. [Effects of the Invention]

[0011] The PPAR activating composition of the present invention contains sucrose fatty acid ester as an active ingredient, and therefore does not affect the taste or flavor, making it suitable for ingestion by adding it to food, and is also highly safe, making it suitable for continuous intake. [Modes for carrying out the invention]

[0012] Next, embodiments for carrying out the present invention will be described. However, the present invention is not limited to these embodiments.

[0013] As mentioned earlier, the PPAR activating composition of the present invention contains sucrose fatty acid ester as an active ingredient. Sucrose fatty acid esters can be obtained by reacting the hydroxyl group of sucrose with the carboxyl group of a fatty acid to form an ester bond. Since there are eight hydroxyl groups in one molecule of sucrose, and 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 constituent fatty acids for the above sucrose fatty acid esters include saturated and unsaturated fatty acids with 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 individually or in combination of two or more. Among these, stearic acid, a saturated fatty acid with 18 carbon atoms, is preferred.

[0014] In the PPAR activating composition of the present invention, the sucrose fatty acid esters described above are used individually or in combination of two or more. In particular, by using two or more different sucrose fatty acid esters in combination, a PPAR activating composition exhibiting performance more suitable for individual requirements can be obtained.

[0015] In the case of the PPAR activating composition of the present invention, for example in liquid form, the sucrose fatty acid ester content is preferably in the range of 0.01 to 20 mg / ml, and more preferably in the range of 0.01 to 10 mg / ml, from the viewpoint of the effectiveness of PPAR activity. Furthermore, the PPAR activating composition of the present invention is not particularly limited in form, such as solid, powder, or liquid, as long as it can be taken orally.

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

[0017] The composition for activating PPAR of the present invention obtained in this way can promote the activation of PPAR by continuously ingesting orally, thereby preventing and improving hyperlipidemia, inflammatory diseases (including skin inflammation), insulin resistance diseases (diabetes, arteriosclerosis, etc.), stroke, circadian rhythm disorder diseases, obesity, promoting the combustion of body fat, reducing blood triglycerides, suppressing fatty liver, improving endurance, promoting the differentiation of epidermal cells and melanocytes (whitening effect by inducing the differentiation of epidermal cells and suppressing the differentiation of melanocytes), promoting keratinization, and promoting the development and repair of the epidermis, etc., and can exert excellent effects.

[0018] In addition, the composition for activating PPAR of the present invention can be directly ingested orally as it is, such as a mixture of sucrose fatty acid ester and soybean powder as described above. However, it is also possible to ingest orally an extract of its active ingredient dissolved or dispersed in a suitable liquid carrier or mixed with a suitable powder carrier.

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

[0020] In addition, when formulating the composition for activating PPAR of the present invention, various components usually used in formulation can be arbitrarily used. Examples thereof include starch, dextrin, lactose, corn starch, inorganic salts, etc. These can be used alone or in combination of two or more.

[0021] Examples of dosage forms during the above formulation include ampoules, tablets, capsules, granules, fine granules, powders, drinkable preparations, etc.

[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 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 exert the PPAR activation effect and the preventive and improvement effects on various diseases related thereto (particularly, obesity and hyperlipidemia, etc.). In addition, not only in humans, but also in animals such as pets and livestock, the above effects can be obtained, and the dosage thereof is appropriately set according to conditions such as the difference in the organisms 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 foods and feeds.

Examples

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

[0025] <PPAR Activation Test> By the following method, an activation test of nuclear receptors (PPARα, PPARδ) was performed by 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 5Cells were seeded in 6-well plates to achieve a cell-per-well ratio and cultured for 1 day in Dulbecco's modified Eagle medium (supplemented with 10% fetal bovine serum (FBS) and 2 mol / l L-glutamine). Next, a chimeric protein expression plasmid (pGal4DBD / NR(LBD)) containing the DNA-binding domain of the Gal4 gene (Gal4-DBD) and the ligand-binding domains (NR-LBD) of various human nuclear receptors (PPARα, PPARδ), a reporter plasmid (pGal4-Luc) containing the Gal4 gene DNA response sequence and the firefly luciferase gene, and an internal standard plasmid (pGL4.75hRluc-CMV; Promega) with a constitutive expression promoter (CMV) linked upstream of the sea urchin luciferase gene were mixed in a weight ratio of 1:0.9:0.1 and dissolved in Opti-MEM medium (GIBCO). After adding the gene transfer reagent X-tremeGENE HP (Roche), the mixture was added to the cultured cells and cultured to introduce the genes. Next, the transgenic cells were detached using trypsin, washed, and then placed in a 96-well plate in a 1.6 × 10⁶ format. 4 The cells were re-seed to a cell / well ratio. At this time, the culture medium was replaced with Dulbecco's modified Eagle medium (phenol red-free, 10% activated carbon treated FBS, SIGMA) containing the test sample, and the cells were cultured for a further 48 hours. The cells were washed with phosphate-buffered saline (PBS), lysed using a dual luciferase assay system (Promega), and then a substrate solution containing luciferin was added. Firefly luciferase activity and sea urchin luciferase activity were measured using a plate reader (Felios AB-2350, ATTO). The PPAR activation test was performed only after confirming that no significant damage was observed to the cells (no significant decrease in sea urchin luciferase activity). The above procedure was performed using three wells per sample (including the negative control), and the mean value of the three wells was used as the 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 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. 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. The product obtained by adding dimethyl sulfoxide (DMSO) to dissolve it to a concentration of 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 the negative control (NC). Note that the evaluation of PPARα activity and PPARδ activity was expressed as the 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] The results in Table 1 above show that the activity of PPARα and PPARδ increases almost proportionally to the concentration of sucrose stearate. Furthermore, even at a small concentration of sucrose stearate (0.04 mg / ml), an increase in the activity of PPARα and PPARδ is observed.

[0030] [Example 2] <Measurement of PPAR activity of soy flour containing sucrose fatty acid esters> An ethanol extract of soy flour containing sucrose stearate was obtained, and this was used as a test sample. The PPAR activity of the test sample was measured in accordance with the method described in Example 1. The evaluation of PPARα activity and PPARδ activity was performed in the same manner as in Example 1. The test samples were prepared as follows: First, two types of sucrose stearate esters (Ryoto Sugar Ester S-370 and Ryoto Sugar Ester S-770, manufactured by Mitsubishi Chemical Foods Co., Ltd.) were mixed in a weight ratio of 1:1. Soybean flour was then added, and the mixture was dissolved in hot water with stirring. Homogenization was then performed to prepare soybean liquid. This soybean liquid was freeze-dried, and the resulting freeze-dried product was mixed with 10 times the volume of 70% ethanol solution and stirred at room temperature for 3 hours. After centrifugation, the supernatant was filtered through a 0.2 μm membrane filter and freeze-dried. This was then dissolved with dimethyl sulfoxide (DMSO) to a concentration of 400 mg / ml and used as the 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 soy 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 soy flour> An ethanol extract of soybean flour was obtained, and this was used as a test sample. The PPAR activity of the test sample was measured in accordance with the method described in Example 1. The evaluation of PPARα activity and PPARδ activity was performed in the same manner as in Example 1. The test samples were obtained using the same method as in Example 2, except that sucrose stearate was omitted. The final concentration of the test sample was adjusted to 2.0 mg / ml of soy 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] As shown in Table 2 above, the activity levels of PPARα and PPARδ in Example 2, in which soy flour was used in combination with sucrose stearate, were significantly improved compared to the activity levels of PPARα and PPARδ when the sucrose stearate concentration was 0.04 mg / ml, as shown in Table 1 above. In particular, a high synergistic effect was observed in the activity of PPARα in Example 2. Furthermore, while Comparative Example 1, which used only soy flour without sucrose stearate, showed a certain degree of high PPARα and PPARδ activity, Example 2 demonstrates that even a small amount of sucrose stearate (sucrose stearate concentration of 0.04 mg / ml) can increase the degree of PPARα and PPARδ activity when used in combination with soy flour.

[0034] Furthermore, the PPAR activity enhancement effect observed in Example 1 and Example 2 above can be considered to be the effect of enhancing PPAR activity when the PPAR activating composition of the present invention is orally ingested as a food product, which is one embodiment of the PPAR activating composition of the present invention. In addition, since the sucrose fatty acid ester, which is the active ingredient of the above composition, does not affect the taste or flavor, it is considered suitable for ingestion by adding it to food products. Moreover, based on the results of the above experiment, it can be expected that the PPAR activating composition of the present invention, when used in combination with soy flour as in Example 2, will yield a higher PPAR activity enhancement effect when orally ingested as food products. [Industrial applicability]

[0035] The PPAR activating composition of the present invention is suitable for ingestion by adding it to food, etc., because its active ingredient, sucrose fatty acid ester, does not affect the taste or flavor, and is also suitable for continuous intake due to its excellent safety. Therefore, the PPAR activating composition of the present invention can be applied as food and beverages, and can also be used as a supplement, pharmaceutical, food additive, etc., by being powdered, tableted, or encapsulated as needed.

Claims

1. A composition for activating peroxisome proliferator-responsive receptors α and δ, characterized by containing a sucrose fatty acid ester as an active ingredient and promoting the activation of peroxisome proliferator-responsive receptors α and δ.

2. The composition for activating peroxisome proliferator-responsive receptors α and δ according to Claim 1, comprising the sucrose fatty acid ester and soy flour.

3. The composition for activating peroxisome proliferator-responsive receptors α and δ according to claim 1 or 2, wherein the constituent fatty acid of the sucrose fatty acid ester is stearic acid.

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