ATP production promoting composition

Abstract

To provide an ATP production promoting composition containing NMN that exhibits enhanced effect in promoting ATP production under oxidative stress.SOLUTION: An ATP production promoting composition contains NMN and at least one selected from CoQ10, resveratrol, and MK-4.SELECTED DRAWING: Figure 4

Description

[Technical field]

[0001] The present invention relates to a composition for promoting ATP production. In particular, the present invention relates to an ATP production-promoting composition that synergistically improves ATP production by combining nicotinamide mononucleotide (NMN) with one or more of coenzyme Q10 (CoQ10), resveratrol, and menaquinone-4 (MK-4). [Background technology]

[0002] NMN (β-nicotinamide mononucleotide) is an intermediate metabolic product of the coenzyme NAD+, which is widely distributed in the body. In recent years, attention has been focused on the relationship between NAD+ and sirtuin (NAD+-dependent deacetylase), which is involved in the aging phenomenon. The amount of NAD+ and sirtuin activity in cells decrease with aging, but it is thought that the aging phenomenon can be suppressed by increasing intracellular NAD+. It is also known that sirtuins can be reactivated by replenishing the body with NAD+ intermediate metabolic products such as NMN. Therefore, the anti-aging effects of NMN are being considered for use in skin care products, cosmetics, food, and other applications. Furthermore, since NMN was added to the "non-drug list" in the 2020 revision of the food and drug classification, its use in health foods, supplements, etc. is expected to expand in the future.

[0003] NAD+ is an essential component in the body and is used in various reactions in the body, such as stress response, gene expression, DNA repair, etc. NAD+ is also used as a coenzyme in glycolysis and the citric acid cycle, and is involved in ATP production in the electron transport chain in mitochondria.

[0004] Patent Document 1 discloses an anti-aging agent containing NMN as an active ingredient, which is shown to reduce active oxygen in the body.

[0005] Patent Document 2 discloses a composition that contains NMN and protects cells from oxidative stress and mitochondrial stress, and indicates that the composition can be used to enhance cell resistance to DNA damage, oxidative stress, and mitochondrial dysfunction, and to improve DNA repair ability.

[0006] Patent Document 3 discloses an NMN-containing antioxidant composition that contains NMN, CoQ10, and vitamins, and describes that NMN can play a role in activating the body's energy metabolism and improving the body's oxidative stress response, and at the same time, due to the synergistic effects with CoQ10, vitamins, etc., it is possible to obtain excellent effects such as antioxidant properties, delayed aging, and improved metabolic rate.

[0007] It is known that NAD+ is involved in ATP production (Non-Patent Document 1), but it was not known that NMN works in concert with other food components to synergistically promote ATP production. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] International Publication WO2020 / 054795 [Patent Document 2] Special Publication No. 2021-524495 [Patent Document 3] JP 2022-54483 A [Non-patent literature]

[0009] [Non-Patent Document 1] Regulation of biological functions by vitamin B3-related molecules, Matsuoka et al., Bulletin of Chiba University of Science, Vol.13, 111-117 2020 Summary of the Invention [Problem to be solved by the invention]

[0010] An object of the present invention is to provide an NMN-containing ATP production-promoting composition that exhibits a superior ATP production-promoting effect under oxidative stress. [Means for solving the problem]

[0011] As a result of extensive research to solve the above problems, the present inventors discovered that combining NMN with CoQ10, resveratrol, or MK-4 works in concert to synergistically promote ATP production, and thus completed the present invention.

[0012] Therefore, the above problems can be solved by the following composition. (1) An ATP production-promoting composition comprising NMN and one or more of CoQ10, resveratrol, and MK-4. (2) The composition according to (1), in which CoQ10 is 1 to 500 mass %, resveratrol is 0.05 to 70 mass %, and MK-4 is 0.01 to 150 mass %, relative to 100 mass % of NMN. (3) The composition according to (1) or (2), which is for oral administration. (4) The composition according to any one of (1) to (3), which promotes ATP production by improving mitochondrial function. (5) The composition according to any one of (1) to (4), wherein the improvement in mitochondrial function is a protective effect against oxidative stress. (6) The composition according to any one of (1) to (5), which acts on brain nerve cells. Effect of the Invention

[0013] The present invention can provide an NMN-containing ATP production-promoting composition that exhibits a superior ATP production-promoting effect under oxidative stress.

[0014] When the NMN in the ATP production-promoting composition of the present invention works in coordination with the health food ingredients CoQ10, resveratrol, and MK-4 to synergistically promote ATP production, it is expected to promote higher energy production and suppress the decline in energy production that accompanies aging, thereby leading to the suppression of the aging phenomenon.

[0015] The main ingredient of the present invention, NMN, has a high market price, so by combining it with cheaper CoQ10, resveratrol, and MK-4, the amount of NMN used can be reduced, making it easier to use from a cost perspective, which is of great industrial significance. [Brief description of the drawings]

[0016] [Figure 1] Shows the effect of combining NMN and CoQ10. [Diagram 2] Shows the effect of combining NMN and resveratrol. [Diagram 3] The effect of combining NMN and MK-4 is shown. [Figure 4] Shows the effect of combining NMN with MK-4, CoQ10 or resveratrol. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] The compositions of the present invention contain NMN and one or more of CoQ10, resveratrol, and MK-4.

[0018] (NMN) The β-nicotinamide mononucleotide used in the present invention is not particularly limited, and various commercially available products can be used. Examples of commercially available products include products manufactured by Luoyang Huarong Biotechnology Co., Ltd. and SyncoZymes. The content of NMN in the ATP production promoting composition of the present invention is not particularly limited, but is preferably 1 to 80% by mass, more preferably 6 to 60% by mass. If it is less than 1% by mass, a large amount must be taken to achieve the desired effect, which is undesirable, and if it exceeds 80% by mass, the flowability of the powder becomes poor when handling it, which is undesirable as it reduces the production yield.

[0019] (CoQ10) Coenzyme Q10, also known as ubidecarenone or coenzyme Q10, is a type of coenzyme present in higher animals. Coenzyme Q10 is known to not only have biological activity as a coenzyme, but also to have effects such as energy production and antioxidant properties. The coenzyme Q10 used in the present invention can be obtained by known means, such as those extracted from a living body, those produced by a synthetic method, those produced by a fermentation method, etc., without any particular limitations. In addition, since coenzyme Q10 is converted to a reduced form when absorbed into a living body, the coenzyme Q10 used in the present invention may be a reduced form. The content of coenzyme Q10 in the ATP production promoting composition of the present invention is not particularly limited, but is preferably 1 to 50% by mass, and more preferably 2 to 20% by mass.

[0020] (Resveratrol) The resveratrol of the present invention is a type of stilbenoid (stilbene derivative) polyphenol, and has the systematic name 3,5,4'-trihydroxy-trans-stilbene. Resveratrol is known as an antioxidant contained in red grape skin and red wine. In the composition of the present invention, resveratrol obtained by known means, such as those extracted from plants or foods, or those produced by synthetic methods, can be used without particular limitation. The content of resveratrol in the ATP production promoting composition of the present invention is not particularly limited, but is preferably 0.1 to 50% by mass, and more preferably 0.3 to 30% by mass.

[0021] (MK-4) Menaquinone-4 (MK-4) is a form of vitamin K that is widely distributed in animal foods and is known to have high blood coagulation activity. In the composition of the present invention, those obtained by known means, such as those extracted from animal or plant tissues and those produced by synthetic methods, can be used without particular limitation. The content of MK-4 in the ATP production promoting composition of the present invention is not particularly limited, but is preferably 0.0001 to 10% by mass, and more preferably 0.0003 to 3% by mass.

[0022] The amounts of active ingredients in the ATP production-promoting composition of the present invention are preferably 1 to 500% by mass of CoQ10, 0.05 to 70% by mass of resveratrol, and 0.01 to 150% by mass of MK-4 relative to 100% by mass of NMN, and more preferably 2 to 50% by mass of CoQ10, 0.3 to 10% by mass of resveratrol, and 0.1 to 15% by mass of MK-4.

[0023] (Oxidative Stress) In this specification, oxidative stress refers to damage to cells and individuals caused by reactive oxygen species such as free radicals (superoxide anion, hydroxyl radical, hydrogen peroxide, singlet oxygen, etc.). Oxidative stress is closely related to the decline of biological functions due to the accumulation of abnormalities at the molecular level (protein, lipid, DNA, etc.) that occurs in the body, and is known to be involved in various diseases.

[0024] (Expected effects and uses from promoting ATP production under oxidative stress) Mitochondria contain many important biochemical systems, including the citric acid cycle, electron transport chain, ATP-producing enzymes, heme synthesis, fatty acid oxidation, and urea cycle, which are central to energy metabolism. It is known that mitochondrial reactive oxygen species production increases with age, and oxidative damage to mitochondria further increases the production of reactive oxygen species in mitochondria, accelerating the rate of damage. ATP produced in the electron transport chain is an important source of energy in the body, but when mitochondria are damaged by oxidative stress caused by aging, it is thought that this can have a negative effect on ATP production. It is expected that promoting ATP production even under oxidative stress will help suppress the decline in biological functions that occurs with aging.

[0025] (Expected effects and uses from effects on brain nerve cells) Dementia is caused by the dysfunction or decrease in the number of brain cells, but in reality it is often caused by the decrease in the number of brain cells due to neuronal death. Amyotrophic lateral sclerosis (ALS) is known as a pathology in which free radicals are clearly involved. It is expected that the action of the ATP production-promoting composition of the present invention on brain nerve cells will lead to the prevention of brain cell dysfunction and a decrease in brain cell number. EXAMPLES

[0026] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples.

[0027] (Examples 1 to 8) (Measurement of ATP production under oxidative stress in mouse hippocampal-derived neuronal cells, HT-22) Mouse hippocampal-derived neuronal cells, HT-22, were obtained from Sigma-Aldrich. 1.25 × 10 HT-22 cells were placed in a 96-well plate. 4 200μL of cells / mL were seeded and cultured at 37℃ in a 5% carbon dioxide atmosphere in DMEM medium (Sigma) containing fetal bovine serum (10%) and antibiotics (100 U / mL penicillin, 100μg / mL streptomycin) for 18 hours. After removing the medium, medium was prepared so that NMN had a final concentration of 0, 10, 100, 1000μM and CoQ10 had a final concentration of 0, 9, 18μM (Examples 1 and 2). Similarly, medium was prepared so that NMN had a final concentration of 0, 10, 100, 1000μM, resveratrol (Tokyo Chemical Industry Co., Ltd.) had a final concentration of 0, 1, 5, 10μM (Examples 3 to 5), or NMN had a final concentration of 0, 10, 100, 1000μM, and MK-4 had a final concentration of 0, 0.1, 1, 10μM (Examples 6 to 8). Each medium was added at 200 μL / well, and after 24 hours of culture, 20 μL of medium adjusted to 200 μM hydrogen peroxide was added to each well. After 1 hour of culture, the amount of intracellular ATP was quantified using the Intracellular ATP Measurement Kit ver.2 (Toyo B-Net Co., Ltd.). The amount of protein in the cells was quantified by the Lowry method. Wells to which neither the test substance nor hydrogen peroxide was added were used as controls, and the amount of ATP per unit protein was calculated from the quantified values ​​obtained, and a relative ratio was calculated with the control set at 1.

[0028] [Table 1]

[0029] (Examples 9 to 11) (Evaluation of mitochondrial oxidative stress under oxidative stress in mouse hippocampal-derived neuronal cells, HT-22) To evaluate mitochondrial oxidative stress, MitoSOX (Invitrogen) was dissolved in DMSO and then diluted with DMEM medium to prepare a concentration of 5 μM. HT-22 cells (3.2 x 10) in a 6-well plate 4 The cells were seeded at 200 μL per mL and incubated at 37°C in a 5% carbon dioxide atmosphere with fetal bovine serum (10%), antibiotics (100 DMEM medium (S The cells were cultured in igma for 18 hours. After removing the medium, 2 μL / well of medium prepared to have a final concentration of NMN of 10 or 100 μM, a final concentration of CoQ10 of 9 μM, a final concentration of MK-4 of 1 μM, and a final concentration of resveratrol of 1 μM was added. After 24 hours of culture, 200 μL of medium prepared to have 200 μM hydrogen peroxide was added to each well, and after 1 hour of culture, the cells were collected by trypsin treatment, washed with PBS, suspended in PBS, and transferred to a 96-well U-shaped plate. After centrifugation to remove the supernatant, 100 μL of the prepared staining reagent was added to each well and stained for 15 minutes at 37 ° C. under a 5% carbon dioxide atmosphere in the dark. The stained cells were collected and the amount of mitochondrial superoxide was measured under PE conditions using a flow cytometer (BD AccuriTM C6 Plus, BD Biosciences).

[0030] It was confirmed that the ATP production promoting composition of the present invention has a more excellent ATP production promoting effect than NMN alone due to the synergistic effect of NMN with CoQ10, resveratrol, or MK-4. In addition, since the amount of mitochondrial superoxide in cells is reduced by the ATP production promoting composition of the present invention, it is presumed that the ATP production promoting effect of the composition of the present invention is largely due to the improvement of mitochondrial function.

[0031] The composition of the present invention can provide an NMN-containing ATP production-promoting composition that has a superior ATP production-promoting effect under oxidative stress, and is therefore highly useful in foods, health foods, pharmaceuticals, etc.

Claims

1. A composition for promoting ATP production comprising NMN and one or more of CoQ10, resveratrol, and MK-4.

2. The composition according to claim 1, wherein CoQ10 is 1 to 500% by mass, resveratrol is 0.05 to 70% by mass, and MK-4 is 0.01 to 150% by mass, relative to 100% by mass of NMN.

3. 3. The composition according to claim 1 or 2, which is for oral administration.

4. The composition of claim 1, which promotes ATP production by improving mitochondrial function.

5. The composition of claim 1 , wherein the improvement in mitochondrial function is a protective effect against oxidative stress.

6. The composition according to claim 1, which acts on brain nerve cells.

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