Cell activators

Abstract

The present invention aims to discover a new function of a composition comprising a combination of nicotinamide mononucleotide (NMN) and a food additive reducing agent derived from shellfish and natural zeolite, and to provide a use based on the new function. [Solution] The cell activator contains nicotinamide mononucleotide (NMN) and a food additive reducing agent obtained by adding baked shell powder obtained by pulverizing baked shells and baked zeolite powder obtained by pulverizing baked natural zeolite to water, subjecting them to a heat treatment in the water, and drying them.

Description

[Technical field]

[0001] The present invention relates to a cell activator containing nicotinamide mononucleotide (NMN) and a reducing agent derived from natural materials, which is prepared by calcining shells and natural zeolite, heating them in water, or the like. [Background technology]

[0002] The sirtuin gene, also known as the anti-aging gene or longevity gene, is known to be activated by the coenzyme nicotinamide adenine dinucleotide (NAD). However, it has been reported that when the amount of NAD in the body decreases due to aging or some other cause, the sirtuin gene is not activated sufficiently, which causes aging. In recent years, nicotinamide mononucleotide (NMN) has been attracting attention. When NMN is taken into the body, it is converted into the coenzyme NAD, which activates the sirtuin gene. Therefore, by taking NMN, it is expected to compensate for the coenzyme NAD that has decreased due to aging, activate the sirtuin gene, and prevent aging.

[0003] In addition to the above-mentioned anti-aging functions, research is being conducted on new functions of compositions containing NMN. For example, Patent Document 1 discloses that a composition combining NMN and leucine has a muscle-building effect. Patent Document 2 discloses that a composition combining NMN and resveratrol has a blood uric acid reducing effect.

[0004] On the other hand, a reducing agent derived from natural materials is known, which is obtained by heating and sintering shells such as oyster shells and natural zeolite in an electric furnace under specified conditions, grinding them into a powder of 10 microns, and then putting both into water, subjecting them to a heat treatment in the water, and drying them (Patent Document 3). This reducing agent for food additives is manufactured and sold as a food ingredient by Wedge Co., Ltd. under the product name "Life Ceramics Powder." [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2022-146639 A [Patent Document 2] Patent No. 6545256 [Patent Document 3] Patent No. 4465687 Summary of the Invention [Problem to be solved by the invention]

[0006] There is a need to discover new functions of NMN that contribute to everyday health and beauty by combining NMN, which has anti-aging functions, with some other material, as disclosed in Patent Documents 1 and 2. It is expected that these new functions will be used to develop functional foods and pharmaceutical compositions.

[0007] Therefore, the present invention has been made in consideration of the above-mentioned points, and its object is to find a new function for a composition comprising a combination of NMN and a food additive reducing agent derived from shells and natural zeolite, and to provide uses based on that new function. [Means for solving the problem]

[0008] The inventors conducted various assays and repeated research on a composition that combines NMN with a food additive reducing agent derived from shellfish and natural zeolite. As a result, the inventors discovered that a composition that combines NMN with a food additive reducing agent has a specific function that is not found in either material alone. Based on this finding, the inventors have completed the present invention.

[0009] In order to solve the above problems, the cell activator of the present invention contains nicotinamide mononucleotide (NMN) and a food additive reducing agent obtained by adding baked shell powder obtained by pulverizing baked shells and baked zeolite powder obtained by pulverizing baked natural zeolite to water, subjecting them to heat treatment in water, and drying them.By making a composition in which NMN and the above-mentioned food additive reducing agent are blended, cells can be activated and their functions can be activated.

[0010] In addition, the cell activator of the present invention preferably activates hepatocytes or fibroblasts. This allows suitable cells to be selected as cells to be activated by the composition of the present invention, which contains NMN and a food additive reducing agent derived from shellfish and natural zeolite. The composition of the present invention can activate both hepatocytes and fibroblasts, thereby activating their functions.

[0011] Furthermore, in the cell activator of the present invention, the blending ratio of nicotinamide mononucleotide to the food additive reducing agent (mass of nicotinamide mononucleotide: mass of food additive reducing agent) is preferably 10:90 to 40:60. This allows the selection of a suitable blending ratio of components that has excellent cell activation effects in the composition of the present invention blended with NMN and the food additive reducing agent.

[0012] In addition, the liver function improver of the present invention contains the above-mentioned cell activator as an active ingredient. The composition according to the present invention can activate liver cells, thereby activating the function of liver cells and improving liver function.

[0013] The skin aging inhibitor of the present invention contains the above-mentioned cell activator as an active ingredient. The composition of the present invention activates fibroblasts, thereby activating the function of fibroblasts and thereby inhibiting or improving skin aging. Effect of the Invention

[0014] According to the present invention, a cell activator having the following excellent effects can be provided. (1) Hepatocytes and fibroblasts can be activated, thereby making it possible to activate the functions of the cells. (2) You can also enjoy the other functions that NMN and food additive reducing agents each possess. (3) Since all of these compounds are highly safe for the human body, they can be taken orally and used safely as functional foods. [Brief description of the drawings]

[0015] [Figure 1] 1 is a graph showing the cell activation effect of the composition of the present invention on hepatocytes in Example 1. Fig. 1(a) shows the test results when the composition of the present invention (Example 1) and only NMN (Comparative Example 1a) were added to hepatocytes, and Fig. 1(b) shows the test results when the composition of the present invention (Example 1) and only a reducing agent (Comparative Example 1b) were added to hepatocytes. [Diagram 2] 2 is a graph showing the cell activation effect of the composition of the present invention on fibroblasts in Example 2. Fig. 2(a) shows the test results when the composition of the present invention (Example 2) and only NMN (Comparative Example 2a) were added to fibroblasts, and Fig. 2(b) shows the test results when the composition of the present invention (Example 2) and only a reducing agent (Comparative Example 2b) were added to fibroblasts. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] The cell activator of the present invention will be described in detail below. The cell activator of the present invention contains nicotinamide mononucleotide (NMN) and a reducing agent for food additives derived from natural materials, which is prepared by calcining shells and natural zeolite, which will be described later, and by heating them in water.

[0017] In the present invention, cell activation refers to an increase in the metabolic activity of cells and an activation of the function of cells, and means that the metabolic activity of cells is improved and the function of cells is activated as compared to a control in which the cell activator of the present invention is not added or administered. Specifically, for example, in an MTT assay, the amount of formazan produced from MTT is increased as compared to a control in which the cell activator of the present invention is not added or administered, that is, the metabolic activity of cells that reduce MTT to formazan is increased.

[0018] The cells that are activated by the composition of the present invention are not particularly limited, but the effect on liver cells or fibroblasts is remarkable. In addition, as fibroblasts, dermal fibroblasts are preferable. Although the mechanism of action by which the composition of the present invention activates these cells has not been elucidated, the composition that combines NMN with a food additive reducing agent derived from shellfish and natural zeolite exhibits excellent cell activation effect.

[0019] Furthermore, the cell activating agent according to the present invention can be used as a liver function improving agent that activates liver cells, thereby activating the metabolism of liver cells and improving liver function. Also, the cell activating agent according to the present invention can be used as a skin aging inhibitor that activates skin fibroblasts, thereby improving the metabolic activity of the skin fibroblasts and suppressing oxidation reactions in the cells, thereby suppressing or improving skin aging.

[0020] (nicotinamide mononucleotide) The composition of the present invention contains nicotinamide mononucleotide (NMN). NMN has α-type and β-type optical isomers, and β-NMN, which is β-type, is preferably used in the present invention. In the present invention, this NMN is combined with a food additive reducing agent described below to improve the metabolic activity of liver cells and skin fibroblasts, and activate these cells. Furthermore, as described above, when NMN is taken into the body, it is converted into the coenzyme NAD, and therefore functions to activate the sirtuin gene. Therefore, by taking NMN, it is expected to have the effect of supplementing the coenzyme NAD that has decreased due to aging, etc., activating the sirtuin gene, and preventing aging.

[0021] (Food additive reducing agent) The composition of the present invention contains a food additive reducing agent obtained by adding baked shell powder obtained by crushing baked shells and baked zeolite powder obtained by crushing baked natural zeolite to water, heat-treating in water, and drying. In the present invention, by combining the above-mentioned NMN with this food additive reducing agent, it has the effect of improving the metabolic activity of liver cells and skin fibroblasts, and activating these cells. As the food additive reducing agent, the product name "Life Ceramics Powder" (product of Wedge Co., Ltd.), which is distributed as a health food material, is preferably used.

[0022] This food additive reducing agent is not particularly limited, but as an example, it can be produced by the method disclosed in Patent Document 3 (Japanese Patent No. 4465687). For example, shells such as oyster shells are placed in an electric furnace and heated and fired for 8 to 20 hours while gradually increasing the temperature in the range of 500°C to 850°C. After heating and firing, the fired shells are ground into powder to obtain fired shell powder. On the other hand, natural zeolite is also heated and fired for 5 to 10 hours using an electric furnace while gradually increasing the temperature in the range of 200°C to 800°C. After heating and firing, the fired natural zeolite is ground into powder to obtain fired zeolite powder. Then, a mixture of 85 to 98% fired shell powder and 2 to 15% fired zeolite powder is added to water, heated to a water temperature of 90°C, and ion-exchanged in water for 5 to 48 hours while maintaining this equilibrium temperature. Thereafter, the mixture is dried to obtain the food additive reducing agent.

[0023] More specifically, shells such as oyster shells are placed in an electric furnace and fired at an initial temperature of 550°C for 3 hours, then at 700°C for 5 hours, and then at 840°C for 8 hours. After firing, the fired shells are crushed into 10 micron powder to obtain fired shell powder. On the other hand, natural zeolite is also fired in an electric furnace at an initial temperature of 250°C for 2 hours, and then at 750°C for 5 hours. After firing, the fired natural zeolite is crushed into 10 micron powder to obtain fired zeolite powder. Then, a mixture of 90% fired shell powder and 10% fired zeolite powder is put into water, heated to a water temperature of 90°C, and ion-exchanged in water for 8 hours while maintaining this equilibrium temperature. After that, the mixture is dried until the water content is 3%, and then fired in an electric furnace at 840°C for 5 hours to obtain the food additive reducing agent. The calcination time and temperature of the shells and natural zeolite, the mixing ratio of the calcined shell powder and the calcined zeolite, and the heating temperature and reaction time in water can be appropriately changed depending on the type and condition of the shells and natural zeolite selected as materials.

[0024] In the composition of the present invention, the compounding ratio of NMN to the food additive reducing agent (mass of NMN: mass of food additive reducing agent) is preferably 10:90 to 40:60, more preferably 15:85 to 30:70, and even more preferably 20:80 to 30:70, from the viewpoint of exerting a cell activation effect. This activates and stimulates hepatocytes and skin fibroblasts.

[0025] In terms of the effects, the amount of NMN in the composition of the present invention is preferably 5-40% by mass, more preferably 10-30% by mass, and particularly preferably 15-30% by mass of the entire composition. In addition, in terms of the effects, the amount of the food additive reducing agent in the composition of the present invention is preferably 50-85% by mass, more preferably 55-80% by mass, and particularly preferably 55-70% by mass of the entire composition.

[0026] The cell activating composition according to the present invention can be produced according to a known method, specifically by mixing and dispersing the above-mentioned components.

[0027] In addition to the above-mentioned ingredients such as NMN and food additive reducing agents, the composition of the present invention can be prepared in various forms by conventional methods by appropriately mixing ingredients such as excipients, lubricants, dispersants, disintegrants, buffers, bulking agents, preservatives, flavors, or stabilizers that are commonly used in the manufacture of foods such as supplements and pharmaceuticals. In addition, in order to improve the bioavailability and stability of the active ingredients in the composition of the present invention, drug delivery systems including formulation techniques such as microcapsules, ultrafine powders, and inclusion using cyclodextrins can also be used.

[0028] Furthermore, the cell activator according to the present invention can be in any form, such as tablets, capsules (hard capsules and soft capsules), granules, suspensions, syrups, and other supplement forms, beverages such as soft drinks, fruit juices, and alcoholic beverages, sweets such as candy, gum, cookies, biscuits, and chocolate, bread, porridge, cereals, noodles, jellies, soups, dairy products, and seasonings. In addition, the composition according to the cell activator of the present invention can be variously combined with other functional ingredients, various amino acids, various vitamins, minerals, oligosaccharides, prebiotics, or other probiotics, within a range that does not affect the efficacy of the active ingredient of the present invention. The composition of the present invention is highly safe and is preferably used as a food composition, and food compositions include supplements, health foods, functional foods, and foods for specified health uses.

[0029] Furthermore, the daily intake of the cell activator according to the present invention will vary depending on the desired effect, age, sex, body weight, etc., but as an example, it is preferable for the NMN to be 20mg to 500mg and the food additive reducing agent to be 100mg to 6g, it is more preferable for the NMN to be 30mg to 300mg and the food additive reducing agent to be 100mg to 3g, and it is even more preferable for the NMN to be 50mg to 250mg and the food additive reducing agent to be 200mg to 2g.

[0030] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples. EXAMPLES

[0031] [Example 1] 1. Examination of cell activation of hepatocytes In this example, using HepG2 cells, a human hepatic cell line, the cell activation effect of the composition according to the present invention, which contains NMN and a food additive reducing agent derived from shellfish and natural zeolite, on hepatic cells was investigated.

[0032] [Preparation of sample solution] β-Nicotinamide Mononucleotide (β-NMN, product of Oriental Yeast Co., Ltd.) and a food additive reducing agent (product name: Life Ceramics Powder, product of Wedge Co., Ltd.) obtained by putting burnt shells and burnt natural zeolite into water, heating them in water, drying them, and then pulverizing them were mixed in a ratio of β-NMN:food additive reducing agent = 23:77. This was diluted with DMEM medium to prepare sample solutions of different concentrations as shown in Table 1 below. On the other hand, as comparative controls, sample solutions in which only β-NMN was diluted with DMEM medium and sample solutions in which only the food additive reducing agent was diluted with DMEM medium were prepared, as shown in Table 1 below.

[0033] [Table 1]

[0034] The cell activation effect of the composition of the present invention on hepatocytes was confirmed by MTT assay, which can measure the metabolic activity of cells. The metabolic activity of cells can be measured by adding MTT (3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) to the target cells and measuring the amount of formazan, a reduction product, by absorbance. MTT was a product of Dojindo Laboratories, Inc. In this test, HepG2 cells, a human liver-derived tumor cell line, were used as live cells, and the HepG2 cells were cultured in DMEM medium at 37°C, in the presence of 5% CO2, and under 100% saturated water vapor conditions.

[0035] First, HepG2 cells were plated in each well of a 96-well microplate at 10 3The wells were seeded with 100 μL of cells / well and cultured in a CO2 incubator for 24 hours. After culture, the supernatant was removed from each well, and the wells were divided into Example 1, Comparative Example 1a, and Comparative Example 1b groups, and 100 μL of sample solution corresponding to each group shown in Table 1 was added to each well. After culture in a CO2 incubator for 24 hours, the supernatant was removed, the medium was replaced with a new DMEM medium (100 μL / well) that did not contain the sample solution, and 10 μL of 0.5 mg / mL MTT was added to each well. After culture for 4 hours, the supernatant was removed, and 100 μL of solubilizing solution (containing 10% SDS in 0.1 M HCl) was added to each well, and the wells were left to stand overnight to solubilize the formazan crystals that had formed. The next day, the absorbance of the formazan solution at 560 nm was measured using a microplate reader (Tecan Corporation product), and the relative value was calculated as the absorbance ratio (% of control) when the absorbance of the formazan solution in the solvent control test plots (test plots No. 1, No. 1a, and No. 1b) containing only DMEM medium was taken as 100. The results are shown in Tables 2, 3, and Figure 1. Table 2 and Figure 1(a) show the absorbance ratio (% of control) based on the concentration of NMN added, and Table 3 and Figure 1(b) show the absorbance ratio (% of control) based on the concentration of a food additive reducing agent added.

[0036] [Table 2]

[0037] [Table 3]

[0038] From the results of FIG. 1 and Tables 2 and 3, when NMN alone (Comparative Example 1a) or the food additive reducing agent alone (Comparative Example 1b) was used, no significant change was observed in the absorbance ratio (%), i.e., the amount of formazan generated by the metabolism of liver cells. In contrast, by adding the composition of the present invention, which combines NMN and the food additive reducing agent, the amount of formazan increased in a concentration-dependent manner, and it was revealed that the metabolic activity of liver cells was improved and liver cells were activated. For example, in the composition prepared in this Example 1, the absorbance ratio was 125% at least when the concentration of NMN was 0.73 mg / mL and the concentration of the food additive reducing agent was 2.44 mg / mL, and the absorbance ratio reached 255% under the conditions of the concentration of NMN was 6.57 mg / mL and the concentration of the food additive reducing agent was 22 mg / mL. In addition, no change was observed in the number of liver cells in each test section, including the solvent control, and no tendency for the number of liver cells to increase by adding the composition of the present invention was observed. In this way, it has become clear that the combination of NMN with a food additive reducing agent produces an effect of improving and activating the metabolic activity of liver cells that is not seen with either component alone.

[0039] [Example 2] 2. Examination of cell activation of fibroblasts In this example, normal human skin fibroblasts were used to examine the cell activation effect on hepatocytes of the composition according to the present invention, which contains NMN and a food additive reducing agent derived from shellfish and natural zeolite.

[0040] The sample solution was prepared in the same manner as in Example 1 described above, and the sample solution containing only NMN was designated Comparative Example 2a, and the sample solution containing only the food additive reducing agent was designated Comparative Example 2b. The confirmation test of the cell activation effect on fibroblasts was performed in the same manner as in Example 1 described above, except that the cell line used was changed from HepG2 cells to normal human skin fibroblasts, and the relative value was calculated as the absorbance ratio (% of control) when the absorbance of the solvent control test group (test group No. 2, No. 2a, No. 2b) containing only DMEM medium was taken as 100. The results are shown in Table 4, Table 5, and Figure 2. Table 4 and Figure 2(a) show the absorbance ratio (% of control) based on the concentration of NMN added, and Table 5 and Figure 2(b) show the absorbance ratio (% of control) based on the concentration of the food additive reducing agent added.

[0041] [Table 4]

[0042] [Table 5]

[0043] The results in FIG. 2 and Tables 4 and 5 show that no significant change was observed when NMN alone (Comparative Example 2a) or food additive reducing agent alone (Comparative Example 2b) was added, and when NMM alone was added, the amount of formazan produced by the metabolism of fibroblasts was slightly increased, but the NMM addition rate in the test group where the absorbance ratio (%) increased was a high concentration of 19.7 mg / mL. In contrast, it was revealed that the amount of formazan increased significantly in a concentration-dependent manner by adding the composition of the present invention, which combines NMN and a food additive reducing agent, and the metabolic activity of fibroblasts was improved and fibroblasts were activated. For example, in the composition prepared in this Example 2, at least when the NMN concentration was 0.73 mg / mL and the food additive reducing agent concentration was 2.44 mg / mL, the absorbance ratio increased significantly to 148%, and under the conditions of an NMN concentration of 6.57 mg / mL and a food additive reducing agent concentration of 22 mg / mL, the absorbance ratio reached 248%. Furthermore, no change was observed in the number of fibroblast cells in each test section, including the solvent control, and no tendency for the number of fibroblast cells to increase due to the addition of the composition of the present invention was observed. Thus, it was revealed that the combination of NMN and a food additive reducing agent produces an effect of improving and activating the metabolic activity of fibroblasts that is not seen with each component alone.

[0044] The present invention is not limited to the above-described embodiments or examples, and various modified forms of design are also included in the technical scope within the scope that does not deviate from the gist of the invention described in the claims. [Industrial Applicability]

[0045] The cell activator of the present invention can activate cells by improving the metabolic activity of hepatocytes and fibroblasts, and can be effectively used in industries such as functional foods and pharmaceuticals.

Claims

1. A cell activator for hepatocytes, comprising: Nicotinamide mononucleotide (NMN), A food additive reducing agent is obtained by adding a calcined shell powder obtained by crushing a calcined shell and a calcined zeolite powder obtained by crushing a calcined natural zeolite into water, subjecting the powder to a heat treatment in water, and drying the resulting product. A hepatocyte activating agent, wherein the cell activation is an action of improving metabolic activity of hepatocytes.

2. The cell activation agent for hepatocytes described in claim 1, characterized in that the blending ratio of the nicotinamide mononucleotide to the food additive reducing agent (mass of nicotinamide mononucleotide:mass of food additive reducing agent) is 10:90 to 40:

60.

3. A liver function improver comprising the hepatocyte activator according to claim 1 or 2 as an active ingredient.

4. A cell activator for skin fibroblasts, comprising: Nicotinamide mononucleotide (NMN), A food additive reducing agent is obtained by adding a calcined shell powder obtained by crushing a calcined shell and a calcined zeolite powder obtained by crushing a calcined natural zeolite into water, subjecting the powder to a heat treatment in water, and drying the powder. A cell activating agent for skin fibroblasts, wherein the cell activation has the effect of improving metabolic activity of skin fibroblasts.

5. A cell activator for skin fibroblasts as described in Claim 4, characterized in that the mixing ratio of the nicotinamide mononucleotide to the food additive reducing agent (mass of nicotinamide mononucleotide:mass of food additive reducing agent) is 10:90 to 40:60.

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