Food composition for promoting NAD+ production containing bergenin

Abstract

To provide a composition containing bergenin that promotes the production of NAD+ and specifically promotes the recovery of NAD+ production, which declines in a living body with aging.SOLUTION: The present invention provides food compositions for promoting NAD+ production, capable of increasing the intracellular expression of NAMPT and NMNAT, enhancing a NAD+ salvage pathway, and promoting NAD+ production, as well as oral agents, food and beverage compositions, and anti-aging food compositions in a living body, containing the food compositions for promoting NAD+ production.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a food composition for promoting NAD+ production that contains bergenin. The present invention relates to a food composition for promoting NAD+ production that is highly safe and can be used as an internal medicine. [Background technology]

[0002] In recent years, while the average life expectancy of the world has continued to increase, especially in developed countries, there is a tendency for the gap between the average life expectancy and the healthy life expectancy to widen. Healthy life expectancy is defined as the period during which one can live without being restricted in daily life by health problems. Reducing the decline in functions that occurs with aging, such as vision, hearing, cognition, motor skills, immune function, and organ function, is highly desirable because it leads to an extension of healthy life expectancy. From this perspective, nicotinamide adenine dinucleotide (hereinafter referred to as NAD+, but sometimes referred to as NAD), an essential coenzyme for energy production in the body, has attracted attention. NAD+ is known to function as a coenzyme for dehydrogenases and is involved in glycolysis, the citric acid cycle, the electron transport system, etc. in the body. NAD+ is also used as a substrate for enzyme reactions catalyzed by sirtuins, poly ADP-ribose polymerases (PARPs), CD38, etc.

[0003] In mammals, the main biosynthesis of NAD+ proceeds via a salvage pathway consisting of two enzymatic steps with nicotinamide (NAM) as the main substrate. In the first step, NAM is converted to nicotinamide mononucleotide (NMN) by nicotinamide phosphoribosyltransferase (NAMPT). In the second step, NMN is synthesized to NAD+ by NMN adenylyltransferase (NMNAT). It is known that the amount of NAD+ in the body, including the skin, decreases with age (see, for example, Non-Patent Document 1). In addition, one of the factors that contribute to the amount of NAD+ in the body is the decline in the activity of NAMPT, an enzyme essential for maintaining the amount of intracellular NAD+, with age (see, for example, Non-Patent Document 2). On the other hand, it is known that increasing NAD+ production in the body, such as by enhancing the expression of the NAMPT gene, extends healthy lifespan (see, for example, Non-Patent Document 3). In addition, it is believed that promoting NAD+ production is effective in treating arteriosclerosis, heart disease, diabetes, neurodegenerative diseases, cognitive disorders, bone aging, stress, and the like. In this way, it is believed that promoting NAD+ production leads to the suppression of various physiological functional declines caused by aging. Generally, NMN and nicotinamide riboside are well known as compounds that promote NAD+ production (see, for example, Non-Patent Document 4). However, there has been a demand for ingredients that are safer, more stable, and industrially available at low cost.

[0004] Bergenin is a type of polyphenol that has been reported to have anti-inflammatory and antioxidant effects. The physiological effects of bergenin have been reported in various previous studies, including its anti-inflammatory effects in mice with colitis and its hepatoprotective effects in mice with liver damage. In particular, there have been many reports on its intestinal regulating effects, and Mallotus japonicus, a deciduous tree of the Euphorbiaceae family that is rich in bergenin, has long been used as a herbal medicine with an intestinal regulating effect. The water-soluble fraction of Mallotus japonicus bark contains 11% to 18% bergenin, and it has been reported that Mallotus japonicus extract has a bowel movement improving effect. While various health benefits of bergenin have been reported, there have been no reports on its NAD+ production effect. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] PLos ONE.Vol.7,e42357(2012) [Non-Patent Document 2] PLos ONE.Vol.12,e0170930(2017) [Non-Patent Document 3] Cell Metabolism.Vol.30, pp.329-342(2019) [Non-Patent Document 4] Front Cell Dev BiolVol.8, fcell.2020.00246(2020) Summary of the Invention [Problem to be solved by the invention]

[0006] To provide a composition containing bergenin that promotes the production of NAD+, and in particular promotes the recovery of the production of NAD+ that decreases in the living body with aging. [Means for solving the problem]

[0007] As a result of extensive research, the inventors have discovered that the use of bergenin and application to a living body can promote the recovery of intracellular NAD+, and have completed the following invention.

[0008] That is, according to one embodiment, the present invention provides a food composition for promoting NAD+ production, which contains bergenin. According to another embodiment, the present invention relates to an internal medicine, a food and beverage composition, and an in vivo anti-aging food composition, which contain the above-described food composition for promoting NAD+ production. Effect of the Invention

[0009] When bergenin is applied, it can significantly promote intracellular NAD+ production, which leads to effects such as cell activation, anti-aging, improved sleep quality, improved cognitive ability, improved bone density, and muscle maintenance. According to the present invention, it is possible to provide a food composition for promoting NAD+ production that contains bergenin. The food composition for promoting NAD+ production can be used as an internal medicine, a food and drink composition, and an anti-aging food composition in the body, and can be very useful for preventing aging in the body. [Brief description of the drawings]

[0010] [Figure 1] Figure 1 shows the increase in intracellular NAD+ in human hepatocytes cultured in the presence of bergenin. A shows the amount of intracellular NAD+ in human-derived HepG2 cells after 8 hours of incubation with 0-2 μM bergenin. B and C show the protein levels of intracellular NAMPT and NMNAT, respectively, after 6 hours of incubation with 0-2 μM bergenin, as determined by immunoblotting. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] Hereinafter, an embodiment of the present invention will be described with reference to the drawings. However, the present invention is not limited to the following specific embodiment, and can be modified as desired within the scope of the technical concept.

[0012] [First embodiment: Food composition for promoting NAD+ production] According to a first embodiment, the present invention relates to a food composition for promoting NAD+ production. The food composition for promoting NAD+ production contains bergenin.

[0013] Bergenin can be extracted from the bark, leaves, roots, etc. of Mallotus japonicus. Synthetic bergenin can also be used. In the present application, a commercially available synthetic product (manufactured by Tokyo Chemical Industry Co., Ltd.) is obtained and used.

[0014] It is also possible to use a Mallotus japonicus extract containing bergenin. In addition to bergenin, various polyphenols such as geraniin, rutin, and corilagin are known to be contained in Mallotus japonicus extract. Mallotus japonicus extract is readily available commercially, for example, Mallotus japonicus extract powder (polyphenol content of 14.0% by mass or more) (all manufactured by Nippon Powder Pharmaceutical Co., Ltd.).

[0015] In order to ensure that bergenin is present stably in the food composition for promoting NAD+ production, the amount of bergenin or the amount of bergenin contained in Mallotus japonicus extract is preferably 0.0001% by mass to 97% by mass, and more preferably 0.001% by mass to 90% by mass.

[0016] In the food composition for promoting NAD+ production according to the present embodiment, the production of NAD+ can be promoted by including bergenin. In addition, the food composition for promoting NAD+ production also has the anti-inflammatory and antioxidant effects that are the conventionally known effects of bergenin.

[0017] The food composition for promoting NAD+ production may contain optional ingredients, including but not limited to placenta extract, collagen peptides, nicotinamide, vitamin C derivatives, arbutin, ellagic acid, kojic acid, tranexamic acid, glycerin, 1,3-butylene glycol, and propylene glycol.

[0018] The collagen peptide contained in the food composition for promoting NAD+ production according to the present embodiment may be a hydrolysate of collagen derived from a living body. More specifically, collagen protein extracted from mammalian tissues such as cows and pigs or fish tissues, such as connective tissues such as skin, bone and tendon, or collagen protein heat-denatured gelatin may be obtained by hydrolysis or the like, or may be artificially synthesized.

[0019] The collagen peptide is preferably a peptide containing at least hydroxyproline (Hyp), and is preferably selected from dipeptides or tripeptides containing Hyp. More specifically, it may be one or a mixture of two or more selected from dipeptides or tripeptides of Ala-Hyp-Gly, Pro-Hyp-Gly, Pro-Hyp, Leu-Hyp, and Ala-Hyp. Among them, it is most preferable to contain the dipeptide Ala-Hyp. Ala-Hyp is preferred because it has a lower molecular weight than tripeptides and is easily absorbed into the body, and is also highly stable among these peptides.

[0020] The food composition for promoting NAD+ production can be obtained by mixing bergenin or Mallotus japonicus extract containing bergenin with the above-mentioned optional ingredients. The obtained food composition for promoting NAD+ production is a liquid or solid composition containing bergenin. The food composition for promoting NAD+ production can be stably stored in a refrigerator for about 1 to 3 years and can promote the production of NAD+ in living cells. In addition, the obtained composition can promote the production of NAD+ even in living cells that have aged and reduced NAD+ production, and therefore can be said to be an NAD+ recovery promoter.

[0021] The food composition for promoting NAD+ production can be added to internal medicines and food and beverage compositions. In particular, it can be added to internal medicines and food and beverage compositions to obtain anti-aging effects. In addition, since the promotion of NAD+ production can provide an anti-aging effect in vivo, a composition containing the food composition for promoting NAD+ production can be used as an anti-aging food composition in vivo.

[0022] Below, we will explain various compositions to which the NAD+ production-promoting food composition can be added.

[0023] [Second embodiment: oral preparation] According to a second embodiment of the present invention, there is provided an internal medicine, which includes the food composition for promoting NAD+ production according to the first embodiment. The internal medicine may be, for example, an oral administration medicine, and examples thereof include tea, capsules, tablets, pills, granules, fine granules, syrup, dry syrup, etc.

[0024] These internal preparations, including the food composition for promoting NAD+ production according to the first embodiment, are manufactured by conventional methods generally known in this field. The internal preparations may contain, for example, starch, lactose, sucrose, mannitol, carboxymethylcellulose, cornstarch, inorganic salts, etc. In preparation, binders, disintegrants, surfactants, lubricants, flow promoters, flavorings, colorants, flavorings, etc. may also be added. For example, when tablets or pills are used, they may be coated with sugar coatings such as sucrose, gelatin, hydroxypropylcellulose, or a film of a gastric or enteric substance, if desired. When the internal preparation is an oral preparation consisting of a liquid composition, it may be a pharmacologically acceptable emulsion, solution, suspension, syrup, etc., and for example, purified water, ethanol, etc. are used as carriers. In addition, auxiliary agents such as wetting agents and suspension agents, sweeteners, flavoring agents, preservatives, etc. may be added as desired.

[0025] The amount of the active ingredient of the food composition for promoting NAD+ production in the oral preparation is, for example, based on the mass of bergenin, preferably 2mg or more, preferably 4mg or more per day for an adult. The upper limit of the dosage is preferably 200mg or less, more preferably 100mg or less per day. Therefore, the food composition for promoting NAD+ production may be contained in the range according to each form so as to achieve the above-mentioned intake amount.

[0026] [Third embodiment: food and beverage composition] According to the third embodiment of the present invention, there is provided a food and drink composition, which includes the food and drink composition for promoting NAD+ production according to the first embodiment. The food and drink composition according to this embodiment can be blended with the food and drink composition for promoting NAD+ production according to the first embodiment together with any optional ingredients that are usually used in food and drink compositions. Examples of such food and drink compositions for humans or animals include breads, confectioneries, noodles, meat products, seafood products, processed grain products, processed vegetables, processed fruits, processed eggs, dairy products, powders, instant confectionery bases, edible oils and fats, soup bases, powdered drinks, seasonings, food additives, beverages, feeds, etc.

[0027] In addition, the food and drink composition includes, for example, nutritional supplements such as vitamins, nutritional supplement drinks, health foods for animals, foods for specified health uses, nutritional functional foods, health functional foods, etc., and can be commercialized in various commonly used forms, such as powders, granules, tablets, capsules, liquids, films, etc. When commercializing, excipients, binders, disintegrants, disintegration inhibitors, absorption promoters, adsorbents, lubricants, colorants, preservatives, fragrances, flavorings, sweeteners, etc. are mixed together with the food composition for promoting NAD+ production according to the first embodiment.

[0028] These food and drink compositions are produced by conventional methods generally known in the art.

[0029] The amount of the active ingredient of the food composition for promoting NAD+ production in such a food and drink composition may be added so as to obtain a desired intake amount based on the mass of bergenin within a range according to the form of the food and drink, and it is preferable to add it to the food and drink within the range of 0.0001% by mass to 97% by mass. For example, it is preferable to add it to liquid food and drink within the range of 0.0001% by mass to 1% by mass, to solid food and drink within the range of 0.5% by mass to 85% by mass, and to tablet food and drink within the range of 1.0% by mass to 97% by mass. More specifically, it is preferable to add it to supplements within the range of 5% by mass to 90% by mass based on the dry weight of Mallotus japonicus powder, although this is not limited thereto.

[0030] As long as the daily intake of Bergenin does not exceed the allowable amount, there is no particular limit to the number of times it can be taken, but it is usually recommended to take it in divided doses several times a day. EXAMPLES

[0031] The present invention will be described in more detail below with reference to examples. However, the present invention is not limited to the following examples.

[0032] (human hepatocyte culture) Human hepatocyte cell line HepG2 (RIKEN Cell Bank) was cultured in Dulbecco's modified Eagle's medium (DMEM, Nissui Pharmaceutical) containing 10% fetal bovine serum (Invitrogen), 100 ng / mL streptomycin, and 100 μg / mL penicillin at 37 °C and 5% CO. 2 The cells were cultured under the following conditions. The medium was changed every other day until the cultured cells reached approximately 50% confluence, and then every day. When the cells reached a density of 80-90%, they were passaged. In all tests, the medium was replaced with serum-free DMEM beforehand. After overnight incubation, the cells were cultured in the presence or absence of bergenin.

[0033] (Quantification of intracellular NAD+) The NAD+ / NADH ratio of HepG2 cells was determined using the Amplite® Colorimetric NAD+ / NADH Ratio Assay Kit (AAT Bioquest). Cells were washed twice with ice-cold phosphate-buffered saline (PBS) and resuspended in 10% HClO 4 After soaking in PBS, the cells were scraped off the culture plate. After transferring to a new tube, the cells were homogenized with an ultrasonic generator Sonicstar85 and left on ice for 10 min. After centrifugation at 16,000 × g for 10 min at 4 °C, the supernatant was added to the NAD+ / NADH reaction mixture and incubated overnight at 4 °C in the dark. After the reaction, the samples were stored as cytoplasmic fractions at -20 °C. The absorbance of the samples was measured at 460 nm using a microplate reader.

[0034] (Immunoblotting) HepG2 cells were lysed in a lysis solution consisting of 150 mM NaCl, 50 mM Tris (pH 7.0), 1 mM ethylenediamine-N,N,N',N'-tetraacetic acid (EDTA), 1% NP-40, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), and a protease inhibitor cocktail (Nacalai Tesque). Proteins in each cell lysate were separated by SDS-polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane using a semi-dry blotting apparatus (ATTO). After transfer, the membrane was blocked for 1 h with PBS containing 0.1% Tween-20 (PBST) containing 20% ​​Blocking One (Nacalai Tesque) to reduce nonspecific binding. The membrane was then incubated with primary antibodies overnight at 4°C, washed, and incubated with horseradish peroxidase-conjugated secondary antibodies for 1 h. Proteins were detected using chemiluminescence reagents (EZ west Lumi One, ATTO) and a fluorescence scanner (Image Quant LAS 4000 (Cytiva). The following primary antibodies were used: anti-β-actin (1:20,000, Santa Cruz Biotechnology, CA, USA), anti-NMNAT (1:4,000, Santa Cruz Biotechnology), and anti-NAMPT (1:4,000, Xavier Technology). Quantification of band intensity was performed using Image J software (National Institutes of Health, Maryland, USA).

[0035] Example 1: Increasing intracellular NAD+ in human hepatocytes To clarify whether bergenin enhances NAD+ production, human HepG2 cells were Cells were incubated with 0-2 μM bergenin. In Fig. 1A, after 8 h of incubation, the amount of intracellular NAD+ was measured using the Amplite® Colorimetric NAD+ / NADH Ratio Assay Kit (AAT Bioquest). The results are the mean ± standard error of triplicate results. In Fig. 1B and C, after 6 h of incubation, the protein levels of intracellular NAMPT and NMNAT were analyzed by immunoblotting. β-Actin was used as a loading control. The intensity of the bands obtained by immunoblotting was analyzed using Image J software. The results are the mean ± standard error of triplicate results. *p<0.05 and **p<0.01 represent the comparison with the bergenin-untreated control (ANOVA, Dunnett's multiple comparison test).

[0036] As shown in Figure 1A, the amount of intracellular NAD+ was significantly increased by adding 2 μM bergenin. This result indicated that bergenin enhances cellular NAD+ production.

[0037] Bergenin is thought to enhance NAD+ production by activating the NAD+ salvage pathway. Therefore, we investigated how bergenin affects the protein expression of NAMPT and NMNAT, two key enzymes in the NAD+ salvage pathway. The results in Figure 1B and C show that bergenin increased the expression of both NAMPT and NMNAT proteins, which are two important enzymes in the NAD+ salvage pathway. These results suggest that bergenin enhances the NAD+ salvage pathway by increasing the expression of NAMPT and NMNAT, thereby activating NAD+ production.

[0038] <Prescription Examples> [Table 1]

[0039] [Table 2] [Industrial Applicability]

[0040] The food composition for promoting NAD+ production of the present invention can provide an internal medicine and a food and drink composition containing the food composition for promoting NAD+ production. It is expected that the internal medicine will have an effect of preventing aging in the body.

Claims

1. A food composition for promoting NAD+ production, comprising 0.0001% by mass to 97% by mass of bergenin as an active ingredient.

2. 2. A food composition for promoting NAD+ production according to claim 1, which is used for cell activation.

3. 2. A food composition for promoting NAD+ production as described in claim 1, which is used to improve sleep quality.

4. The food composition for promoting NAD+ production according to claim 1, characterized in that it is used for improving cognitive ability.

5. 2. A food composition for promoting NAD+ production as described in claim 1, which is used to improve bone density.

6. A food composition for promoting NAD+ production as described in claim 1, characterized in that it is used for maintaining muscle strength.

7. An oral preparation comprising a food composition for promoting NAD+ production according to any one of claims 1 to 6, which contains bergenin in an amount of 0.0001% by mass to 1% by mass for liquid foods and beverages, 0.5% by mass to 85% by mass for solid foods and beverages, and 1.0% by mass to 97% by mass for tablet-type foods and beverages.

8. A food and beverage composition comprising the food and beverage composition for promoting NAD+ production described in any one of claims 1 to 6, which contains bergenin in an amount of 0.0001% by mass to 1% by mass for liquid foods and beverages, 0.5% by mass to 85% by mass for solid foods and beverages, and 1.0% by mass to 97% by mass for tablet-type foods and beverages.

9. 7. An anti-aging food composition for an in vivo organism, comprising a food composition for promoting NAD+ production according to any one of claims 1 to 6, which contains bergenin in an amount of 0.0001% by mass to 1% by mass for liquid foods and beverages, 0.5% by mass to 85% by mass for solid foods and beverages, and 1.0% by mass to 97% by mass for tablet-type foods and beverages.

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