Autophagy Activation Composition

Abstract

The object of the present invention is to provide a composition having novel autophagy-activating ability, as well as a method for activating autophagy. [Solution] By using a composition containing nicotinamide adenine dinucleotide or a nicotinamide adenine dinucleotide precursor, autophagy in vivo can be activated. Since autophagy has been linked to lifestyle-related diseases and age-related diseases, activating autophagy through this invention can contribute to extending healthy life expectancy.

Description

Technical Field

[0001] The present invention relates to a composition for activating autophagy containing nicotinamide adenine dinucleotide or a nicotinamide adenine dinucleotide precursor.

Background Art

[0002] Autophagy is a mechanism by which cells decompose and metabolize intracellular proteins and organelles, and is a mechanism observed in eukaryotes from yeast to humans. Autophagy contributes to maintaining the homeostasis of the living body by suppressing the accumulation of abnormal proteins in cells and recycling proteins when the nutritional environment deteriorates. In addition, autophagy has been reported to be involved in various diseases, and the relationship between the decrease in autophagy activity and lifestyle-related diseases and age-related diseases such as neurodegenerative diseases, immune diseases, inflammatory diseases, respiratory diseases, kidney diseases, infectious diseases, and diabetes is known.

[0003] As methods for activating autophagy in vivo, methods using ascorbic acid and cysteine (Patent Document 1), methods using D-tryptophan or D-aspartic acid (Patent Document 2), and methods using ergothioneine (Patent Document 3) have been reported. However, the ability to activate autophagy in vivo is still insufficient, and a composition having a more excellent autophagy activation ability has been demanded.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Patent Document 2

Patent Document 3

Non-Patent Documents

[0005] [Non-Patent Document 1] Autophagy. 452-460. (2007) [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] The object of the present invention is to provide a composition having novel autophagy-activating ability and a method for activating autophagy. [Means for solving the problem]

[0007] As a result of diligent research into the above-mentioned problems, the present inventors discovered that nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide precursor exhibits autophagy-activating ability, and thus completed the present invention.

[0008] In other words, the present invention relates to the following (1) to (7). (1) A composition for activating autophagy, comprising nicotinamide adenine dinucleotide or a nicotinamide adenine dinucleotide precursor. (2) The autophagy activating composition according to claim 1, wherein the nicotinamide adenine dinucleotide precursor is one or more selected from the group consisting of nicotinamide mononucleotide, nicotinamide riboside, nicotinamide, and nicotinic acid mononucleotide. (3) The composition described in (1) is an autophagy activating composition, which is a yeast extract containing nicotinamide adenine dinucleotide or a nicotinamide adenine dinucleotide precursor. (4) A method for activating autophagy, characterized by using any of the compositions described in (1) to (3) as an oral or topical preparation. (5) A food or beverage containing any of the compositions described in (1) to (3). (6) A pharmaceutical product comprising any of the compositions described in (1) to (3). (7) A cosmetic product containing any of the compositions described in (1) to (3). [Effects of the Invention]

[0009] According to the present invention, autophagy in vivo can be activated by using a composition containing nicotinamide adenine dinucleotide or a nicotinamide adenine dinucleotide precursor. Since autophagy has been linked to lifestyle-related diseases and age-related diseases, the present invention can extend healthy life expectancy. [Modes for carrying out the invention]

[0010] The composition of the present invention comprises nicotinamide adenine dinucleotide or a nicotinamide adenine dinucleotide precursor. Here, nicotinamide adenine dinucleotide refers to the compound indicated by CAS number 53-84-9 and its salts. Furthermore, the nicotinamide adenine dinucleotide precursor in the present invention refers to a compound that produces nicotinamide adenine dinucleotide through one or two metabolic steps in vivo, such as nicotinamide mononucleotide (CAS number: 1094-61-7), nicotinamide riboside (CAS number: 1341-23-7), nicotinamide (CAS number: 98-92-0), nicotinic acid mononucleotide (CAS number: 321-02-3), and nicotinic acid adenine dinucleotide (CAS number: 6450-77-7), as well as their salts. The composition of the present invention may also use a combination of nicotinamide adenine dinucleotide and a nicotinamide adenine dinucleotide precursor.

[0011] As the nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide precursor included in the composition of the present invention, generally available materials can be used. In addition, generally available materials containing nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide precursor may be used as the composition of the present invention. For example, "NMN faIne" (manufactured by Mitsubishi Corporation Life Sciences), a composition containing nicotinamide mononucleotide, may be used as the composition of the present invention.

[0012] Furthermore, the composition of the present invention may also use a yeast extract containing nicotinamide adenine dinucleotide or a nicotinamide adenine dinucleotide precursor. In this case, the composition of the present invention may be a yeast extract obtained by conventional methods from yeast containing nicotinamide adenine dinucleotide or a nicotinamide adenine dinucleotide precursor. The type of yeast is not particularly limited, but budding yeast (Saccharomyces cerevisiae), fission yeast (Schizosaccharomyces pombe), and Torula yeast (Schizobellindonella jadini) that have a history of being consumed are preferred.

[0013] The nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide precursor contained in the composition of the present invention can be quantified by general analytical methods, such as quantitative analysis using high-performance liquid chromatography.

[0014] The composition of the present invention possesses autophagy-activating ability. Autophagy-activating ability can be determined by measuring the autophagy flux according to the method described in previously published papers (e.g., Non-Patent Literature 1), for example, as follows: Cells stably expressing LC3 (tfLC3; RFP-GFP-LC3), in which the fluorescent proteins RFP and GFP are linked in tandem, are seeded in a 96-well plate and cultured in a 5% CO2 incubator for 24 hours, after which the test substance is added and cultured for another 24 hours. A test group without the test substance is used as a negative control, and the autophagy-activating ability of the test substance can be quantified and evaluated from the GFP / RFP value in the test substance-added group, when the ratio of fluorescence intensities of GFP and RFP in the negative control (GFP / RFP) is set to 1. For example, the smaller the GFP / RFP value compared to the negative control, the higher the autophagy activity, i.e., the higher the autophagy-activating ability.

[0015] The composition of the present invention can be used orally or externally. When used orally, the composition of the present invention can be used as a food or beverage or supplement, or as an orally administered pharmaceutical. When used as a food or beverage, supplement, or pharmaceutical, the composition of the present invention can be mixed with other materials, etc., as long as it does not inhibit the effects of the present invention. In addition, other substances can be added as long as they do not inhibit the effects of the present invention. Examples of substances that can be added include excipients that are commonly added to food and beverages, supplements, and pharmaceuticals, various vitamins, inorganic substances such as calcium, magnesium, iron, and zinc, organic acids such as lactic acid, citric acid, and malic acid, proteins, lactic acid bacteria extracts, plant extracts, and other organic substances, which can be added as appropriate. Acidulants, sweeteners, flavorings, and stabilizers that are commonly added to general foods can also be added. Furthermore, when the composition of the present invention is used as an oral pharmaceutical, the dosage form is not particularly limited and may be any of the following: tablets, powders, granules, pills, suspensions, emulsions, infusions, decoctions, capsules, syrups, liquids, elixirs, extracts, tinctures, fluid extracts, etc. These preparation methods can be conventional methods and are not limited in this invention.

[0016] Furthermore, when the composition of the present invention is used externally, it can be used by applying it to the body surface as a topical pharmaceutical or cosmetic. When the composition of the present invention is used as a topical pharmaceutical or cosmetic, it can be used as is or mixed with other materials. In addition, other substances can be added as long as they do not inhibit the effects of the present invention. Examples of substances that can be added include excipients commonly added to cosmetics and topical pharmaceuticals, various vitamins, inorganic substances such as calcium, magnesium, iron, and zinc, organic acids such as lactic acid, citric acid, and malic acid, proteins, lactic acid bacteria extracts, plant extracts, and other organic substances, which can be added as appropriate. When the composition of the present invention is used as a topical preparation, the dosage form is not particularly limited and may be a cream, liquid, or paste. Conventional methods can be used for these preparations, and there are no limitations in the present invention.

[0017] The dosage of the present invention is not particularly limited as long as it can increase the amount of nicotinamide adenine dinucleotide in the body, and can be set based on the general daily intake of nicotinamide adenine dinucleotide or whole nicotinamide adenine dinucleotide. The intake and frequency of administration to humans will vary depending on the form of administration, the age and weight of the recipient, etc., but for example, in the case of nicotinamide mononucleotide, which is a precursor of nicotinamide adenine dinucleotide, adults should take nicotinamide mononucleotide once to several times a day, preferably 100 mg or more, more preferably 200 mg or more, even more preferably 250 mg or more, and 500 mg or less per day. When using yeast extract containing nicotinamide mononucleotide, measure the nicotinamide mononucleotide content in the yeast extract and adjust the intake to match the dosage in this paragraph. In order to make the daily intake 500 mg or less, the content in the composition is preferably also 500 mg or less. Also, in the case of, for example, the form of tablets, the content per tablet of the composition of the present invention may be 250 mg, or, for example, the content per tablet may be 50 mg, and five tablets may be the composition of the present application. The same applies to granules, liquids, and the like.

[0018] The composition of the present invention can be used not only as a pharmaceutical but also as a food or drink or a cosmetic. In that case, it may be used such that the amount of nicotinamide adenine dinucleotide or a nicotinamide adenine dinucleotide precursor is within the range of the intake amount in the previous stage. Further, the composition of the present invention can be used for activating autophagy.

[0019] By using the composition of the present invention by the method described up to the previous stage, autophagy in the living body can be activated, and the homeostasis in the living body can be maintained.

Examples

[0020] Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited thereto.

[0021] <Sample> As a sample, generally available reagent grade nicotinamide adenine dinucleotide (manufacturer: FUJIFILM Wako Pure Chemical Corporation) was used.

[0022] <Method for measuring autophagy activity> According to the method described in Non-Patent Document 1, the autophagy activity was evaluated. Specifically, HeLa cells stably expressing tfLC3 (RFP-GFP-LC3) were seeded in a 96-well plate and cultured for 24 hours in D-MEM (manufacturer: all Sigma Aldrich) supplemented with 10% FBS and 2 mM L-glutamine in a 5% CO2 incubator. Subsequently, nicotinamide adenine dinucleotide was dissolved in 0.1% DMSO and added to a culture medium to a final concentration of 0.03 mM (Example 1), 0.3 mM (Example 2), or 1 mM (Example 3), or a culture medium containing only 0.1% DMSO (Comparative Example 1), and the cells were incubated for a further 24 hours. After culturing, the fluorescence intensity derived from RFP and GFP in each test group was measured, and the GFP / RFP ratio was calculated for Comparative Example 1 and Examples 1-3. Then, the GFP / RFP ratio for Examples 1-3 was set to 1. Two test groups were prepared on one plate for each test group, and a total of three plates (n=3) were used for the test.

[0023] <Evaluation of autophagy activity> The results of autophagy measurements are shown in Table 1. When the GFP / RFP value for Comparative Example 1 is set to 1, the GFP / RFP value decreases when autophagy is activated, and conversely, increases when autophagy is suppressed. The evaluation results showed that all three examples (1-3) exhibited lower GFP / RFP values ​​compared to Comparative Example 1. This indicates that nicotinamide adenine dinucleotide activates autophagy. Furthermore, autophagy activation was observed even at a low concentration of 0.03 mM, and the decrease in GFP / RFP values ​​was concentration-dependent, indicating that autophagy was more strongly activated. Furthermore, nicotinamide adenine dinucleotide precursors, which are converted to nicotinamide adenine dinucleotide through metabolism in the body, are thought to exhibit similar effects.

[0024] [Table 1]

[0025] From the above, it has been shown that the composition of the present invention, which contains nicotinamide adenine dinucleotide, has autophagy-activating ability. It is believed that it functions similarly in vivo and can activate autophagy. Furthermore, it is believed that nicotinamide adenine dinucleotide precursors that are converted to nicotinamide adenine dinucleotide through metabolism in vivo have a similar effect. Moreover, foods, pharmaceuticals, and cosmetics containing the composition of the present invention can be used as substances that have autophagy-activating ability.

Claims

1. A composition for activating autophagy, comprising nicotinamide adenine dinucleotide or a nicotinamide adenine dinucleotide precursor.

2. The autophagy activating composition according to claim 1, wherein the nicotinamide adenine dinucleotide precursor is one or more selected from the group consisting of nicotinamide mononucleotide, nicotinamide riboside, nicotinamide, and nicotinic acid mononucleotide.

3. A composition for activating autophagy, wherein the composition according to claim 1 is a yeast extract containing nicotinamide adenine dinucleotide or a nicotinamide adenine dinucleotide precursor.

4. A method for activating autophagy, characterized by using the composition described in any one of claims 1 to 3 as an oral or topical preparation.

5. Food and beverages comprising the composition according to any one of claims 1 to 3.

6. A pharmaceutical product comprising the composition described in any one of claims 1 to 3.

7. A cosmetic comprising the composition described in any one of claims 1 to 3.

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