Combination of NMN and euphoria longana lamarck

Abstract

To provide a combination of NMN and Euphoria longana Lamarck.SOLUTION: The present invention provides a combination of NMN and Euphoria longana Lamarck.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to the combination of NMN and Longan. [Background technology]

[0002] Today, many people suffer from symptoms of fatigue, frailty, and lifestyle-related diseases (diabetes, etc.) associated with aging of the whole body, including aging of the skin (sagging and wrinkles). One approach to improving such symptoms is to take ingredients that can improve one's constitution, such as nicotinamide adenine dinucleotide and / or its precursor, nicotinamide mononucleotide (NMN). Nicotinamide adenine dinucleotide is a coenzyme that is widely distributed in the body and is used in energy metabolism and is known to be involved in many signal transduction functions. It is believed that a deficiency of nicotinamide adenine dinucleotide causes a decrease in cell function, so it is expected that taking nicotinamide adenine dinucleotide and / or NMN will lead to an improvement in one's constitution.

[0003] In this way, in addition to directly ingesting nicotinamide adenine dinucleotide and / or NMN, it is also possible to improve one's constitution by increasing the amount of nicotinamide adenine dinucleotide and / or NMN in the body by activating factors related to the production of nicotinamide adenine dinucleotide and / or NMN. For example, nicotinamide phosphoribosyltransferase (NAMPT) is a commonly known factor related to the production of nicotinamide adenine dinucleotide and / or NMN. NAMPT is an enzyme involved in the production of NMN, and therefore NAMPT activators and NAMPT production promoters can be used as ingredients to promote the production of nicotinamide adenine dinucleotide and / or NMN.

[0004] In fact, research and development of health foods using nicotinamide adenine dinucleotide, NMN, and factors related to the production of NMN have been conducted for some time. For example, Patent Document 1 discloses a sirtuin activator containing NMN and black turmeric, and a health food composition containing this sirtuin activator (Patent Document 1).

[0005] Thus, various health foods using nicotinamide adenine dinucleotide, NMN, and factors related to its production have been discovered, but their effectiveness is not always sufficient, and there is still a demand for more effective NAMPT activators and NAMPT production promoters, especially those with excellent effects. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Patent Publication No. 2023-0059 Summary of the Invention

[0007] The present inventors have found that a combination of NMN and longan is useful for promoting the production of NAMPT. The present invention is based on this finding.

[0008] Thus, the present invention provides a combination of NMN and longan.

[0009] According to the present invention, the following inventions are provided. (1) A composition comprising NMN for use in combination with longan. (2) A composition comprising longan for use in combination with NMN. (3) A composition comprising NMN and longan. (4) The composition according to any one of (1) to (3) for whitening the skin. (5) A composition according to any one of (1) to (4) for promoting NAMPT production. (6) The composition according to any one of (1) to (5) for promoting production of nicotinamide adenine dinucleotide. (7) The composition according to any one of (1) to (6), wherein CA / CB is 0.2 to 20, where CA is the dry weight of NMN in the composition and CB is the dry weight of longan. (8) The composition according to any one of (1) to (8), which is a food composition. (9) Use of a combination of NMN and longan for the manufacture of a medicament for skin whitening. (10) Use of a combination of NMN and longan for the manufacture of a medicament for promoting the production of NAMPT. (11) Use of a combination of NMN and longan for the manufacture of a medicament for promoting the production of nicotinamide adenine dinucleotide. (12) The use according to any one of (9) to (11), wherein, in the pharmaceutical preparation, the dry weight of NMN is CA and the dry weight of longan is CB, the CA / CB is 0.2 to 20. (13) Use of a combination of NMN and longan for skin whitening. (14) The use of a combination of NMN and longan to promote the production of NAMPT. (15) Use of a combination of NMN and longan to promote the production of nicotinamide adenine dinucleotide. (16) The use according to any one of (13) to (15), wherein CA / CB is 0.2 to 20, where CA is the dry weight of NMN in the composition and CB is the dry weight of longan. (17) Combination of NMN and longan for skin whitening. (18) Combination of NMN and longan to promote the production of NAMPT. (19) The combination of NMN and longan to promote the production of nicotinamide adenine dinucleotide. (20) The combination according to any one of (17) to (19), used so that CA / CB is 0.2 to 20, where CA is the dry weight of NMN and CB is the dry weight of longan. (21) A method for skin whitening in a subject, comprising administering an effective amount of a combination of NMN and longan. (22) A method for promoting the production of NAMPT in a subject, comprising administering an effective amount of a combination of NMN and longan. (23) A method for promoting the production of nicotinamide adenine dinucleotide in a subject, comprising administering an effective amount of a combination of NMN and longan. (24) The method according to any one of (21) to (23), wherein the dry weight of NMN is CA and the dry weight of longan is CB, the CA / CB is administered so that the ratio is 0.2 to 20.

[0010] According to the present invention, a combination of NMN and longan is provided.By using the combination of the present invention, it is also possible to effectively promote the production of NAMPT.In addition, by using the combination of the present invention, it is also possible to effectively promote the production of nicotinamide adenine dinucleotide. [Brief description of the drawings]

[0011] [Figure 1] FIG. 1 shows the results of evaluating the amount of NAMPT contained in the culture medium collected 10 to 12 days after the start of culturing a skin model using a culture medium supplemented with ascorbic acid and / or β-NMN. [Diagram 2] FIG. 2 shows the results of evaluating the amount of NAMPT contained in the culture medium collected 10 to 12 days after the start of culturing the skin model using culture medium supplemented with β-NMN and / or longan cherries. [Diagram 3] FIG. 3 shows the results of evaluating the amount of melanin in a skin model 3 weeks after the start of culture using a culture medium supplemented with ascorbic acid and / or β-NMN. [Figure 4] FIG. 4 shows the results of evaluating the amount of melanin in a skin model 3 weeks after the start of culture using a culture medium containing β-NMN and / or longan cherries. [Diagram 5] FIG. 5 shows the results of evaluating the amount of NAMPT in a skin model 3 weeks after the start of culture using a culture medium supplemented with ascorbic acid and / or β-NMN. [Figure 6] FIG. 6 shows the results of evaluating the amount of NAMPT in a skin model 3 weeks after the start of culture using a culture medium containing β-NMN and / or longan cherries. [Figure 7] FIG. 7 shows the results of evaluating the amount of NAD+ in a skin model 3 weeks after the start of culture using a culture medium supplemented with ascorbic acid and / or β-NMN. [Figure 8] FIG. 8 shows the results of evaluating the amount of NAD+ in a skin model 3 weeks after the start of culture using a culture medium containing β-NMN and / or longan cherries. Description of the Invention

[0012] According to one embodiment of the present invention, there is provided NMN for use in combination with longan or a composition comprising longan. Such NMN may be a composition comprising NMN.

[0013] According to another embodiment of the present invention, there is provided longan for use in combination with NMN or a composition comprising NMN. Such longan may be a composition comprising longan.

[0014] According to yet another embodiment of the present invention, a composition comprising NMN and longan is provided.

[0015] In the present invention, "longan" refers to Euphoria longana Lamarck, a subtropical plant native to southern China, belonging to the genus Euphoria and the family Sapindaceae. The parts of longan used in the present invention are not limited to, but may include arils, seeds, etc., and are preferably arils. The arils of longan are also called longan cherries. Dried longan cherries are commonly used as herbal medicines, especially in China and Japan, and contain glucose, organic acids, vitamin A, vitamin B1, essential oil components such as α-camphene and α-pinene, and fat components such as myristic acid and myristicin, and have been reported to have a nutritious, sedative, and anti-insomnia effect. In the present invention, longan may be used in the form of a dried product of longan cherries, or may be extracted by an appropriate method and used as an extract.

[0016] The method for producing the extract of Longan in the present invention is not particularly limited, and can be produced according to the method commonly used in the art, for example, by extracting the raw material of the extract. The extraction method is not limited, but includes, for example, hot water extraction, ultrasonic extraction, filtration, reflux extraction, solvent (e.g., ethanol) extraction, etc. These may be carried out alone, or two or more methods may be used in combination. In addition, the extract may be further extracted one or more times by the same method to obtain a high-purity extract.

[0017] The type of solvent used for producing the Longan extract of the present invention is not particularly limited, and any solvent known in the art may be used as long as the extract has the desired effect of the present invention. Examples of such solvents include, but are not limited to, water, alcohols having 1 to 4 carbon atoms, ethyl acetate, acetone, chloroform, etc., and two or more of these may be used in combination.

[0018] In the present invention, "NMN" refers to nicotinamide mononucleotide (CAS registration number 1094-61-7), which is known as an intermediate metabolic product in the biosynthesis of the coenzyme nicotinamide adenine dinucleotide. There are still many unknowns regarding the mechanism of uptake and mechanism of action of NMN in the body, and various researches are still being conducted. For example, it is known that NMN has an effect on improving mitochondrial function decline associated with aging, diseases associated with aging, anti-aging effects, etc.

[0019] According to a preferred embodiment of the present invention, the composition of the present invention is a composition for skin whitening. In the present invention, "skin whitening" means increasing the lightness of skin whose lightness has been reduced due to an excess of pigments such as melanin, or maintaining the lightness of skin at a certain level. "Skin whitening" in the present invention may be achieved by, but is not limited to, inhibiting melanin production, and may be achieved, for example, by improving symptoms (such as age spots and freckles) that begin with an increase in melanin.

[0020] The composition of the present invention may be applied in vivo or in vitro.When the composition of the present invention is applied to a target, the target may include, but is not limited to, human beings, primates including chimpanzees, pet animals such as dogs and cats, livestock animals such as cows, horses, sheep and goats, and mammals such as rodents such as mice and rats.

[0021] According to a preferred embodiment of the present invention, the composition of the present invention is a composition for promoting NAMPT production. In the present invention, "NAMPT" is also called nicotinamide phosphoribosyltransferase, and refers to an enzyme that reversibly catalyzes the reaction of producing NMN from nicotinamide and 5-phosphoribosyl-1-pyrophosphate (PRPP). The promotion of NAMPT production in the present invention may be performed either in vivo or in vitro.

[0022] In addition, it is known that the increased production of NAMPT in the body activates physical functions and extends life span, exerting a so-called anti-aging effect (see Yoshida et al., 2019, Cell Metabolism 30, 329-342, etc.). Therefore, according to another preferred embodiment of the present invention, the composition of the present invention is a composition for anti-aging.

[0023] According to a preferred embodiment of the present invention, the composition of the present invention is a composition for promoting the production of nicotinamide adenine dinucleotide (CAS registration number 53-84-9). Nicotinamide adenine dinucleotide in the present invention may be either or both of an oxidized form (NAD+) and a reduced form (NADH), but is preferably NAD+. NAD+ in the present invention is also written as NAD. Promotion of the production of nicotinamide adenine dinucleotide in the present invention may be performed either in vivo or in vitro.

[0024] Generally, NAD+ is not permeable to cell membranes and has no transporters in mammals, including humans, so it cannot be directly absorbed or taken in as a nutrient. Therefore, NAD+ is biosynthesized in vivo using tryptophan, niacin, and other materials. The biosynthetic pathways of NAD+ include the de novo pathway (also called the kynurenine pathway) using tryptophan as a starting material, and the salvage pathway using vitamin B3 (niacin), etc., and the NAD+ biosynthesized in this way acts as an electron acceptor in the glycolysis and Krebs cycle in vivo. The NAD+ in the present invention is synonymous with the commonly known NAD+.

[0025] On the other hand, NADH is biosynthesized in the glycolysis and Krebs cycle in vivo and acts to generate ATP by oxidative phosphorylation in the electron transport system. NADH in the present invention is synonymous with the commonly known NADH.

[0026] The amounts of NMN and longan used in the composition of the present invention can be appropriately determined depending on the application, but preferably, when the dry weight of NMN in the composition is CA and the dry weight of longan is CB, CA / CB is 0.2 to 20, more preferably 0.4 to 10, and more preferably 0.66 to 6.66.

[0027] If NMN or longan is commercially available, it may be used as long as it provides a composition having the desired effect of the present invention.

[0028] The composition of the present invention may be either an oral composition or a topical composition.The NMN and longan in the present invention are preferably absorbed from the digestive tract by oral ingestion, and the active ingredients are believed to act in the body.Therefore, according to one preferred embodiment of the present invention, the composition of the present invention is an oral composition.

[0029] When the composition of the present invention is used as an oral composition, it can further contain known components (additives) that are used in oral compositions.Such components are not particularly limited, but include, for example, emulsifiers, hydrating agents, solvents, emollients, stabilizers, thickeners, preservatives, lubricants, chelating agents, fillers, excipients, powders, fragrances, flavorings, absorbents, dyes, opacifiers, antioxidants, preservatives, vitamins, amino acids, nutrients, minerals (electrolytes), flavorings such as synthetic flavorings and natural flavorings, pectinic acid and its salts, alginic acid and its salts, organic acids, pH regulators, glycerin, alcohol, carbonation agents used in carbonated drinks, etc.

[0030] The oral composition of the present invention may be any of liquid, semi-solid, and solid.

[0031] When the composition of the present invention is used as a topical composition, it can further contain known components (additives) that are used in topical compositions.Such components include, but are not limited to, emulsifiers, hydrating agents, solvents, emollients, stabilizers, thickeners, preservatives, lubricants, chelating agents, fillers, excipients, powders, fragrances, flavorings, absorbents, dyes, opacifiers, antioxidants, vitamins, amino acids, etc.

[0032] The topical compositions of the present invention may be liquid, semi-solid, or solid.

[0033] The composition of the present invention may be any of a pharmaceutical composition, a cosmetic composition, and a food composition, and is preferably a food composition. EXAMPLES

[0034] The present invention will be described in detail with reference to the following examples, but the present invention is not limited to these examples. Contents are expressed in mass % unless otherwise specified.

[0035] Preparation of drugs to be applied alone to the skin model As test substances, β-NMN (manufactured by Oriental Yeast Co., Ltd.) and Longan Niku Extract Powder (manufactured by Matsuura Pharmaceutical Co., Ltd.) were prepared so that their concentrations in the culture medium were 300 μg / mL, 100 μg / mL, and 30 μg / mL, respectively. As a control substance, L-(-)ascorbic acid (manufactured by Kanto Chemical Co., Ltd.) was prepared so that its concentration in the culture medium was 100 μg / mL.

[0036] Preparation of drugs to be applied in combination to the skin model Both β-NMN and Longan Niku extract powder were prepared so that their concentrations in the culture medium were 50μg / mL.In addition, both β-NMN and L-(-)ascorbic acid were prepared so that their concentrations in the culture medium were 50μg / mL.

[0037] Cultivation of skin models A melanocyte-containing three-dimensional skin model (MEL-300, Kurabo Co., Ltd.) was purchased and cultured in a 37°C 5% CO2 incubator according to standard methods. Four wells (n=4) were designed for each condition. EPI-100NMM113 was used as the culture medium, and bFGF, α-MSH, and KGF were added in appropriate amounts as melanin stimulating factors. The culture medium was replaced every other day, and the drug (test substance or control substance) was dissolved to a specified concentration each time the culture medium was replaced. The culture was performed so that each drug was applied from the bottom side of the skin model. The culture was continued for 3 weeks after the start of culture.

[0038] Evaluation of the effects of NMN and longan on skin models First, the amount of NAMPT secreted in the culture medium collected 10 to 12 days after the start of culture was measured using the CircuLex Human NAMPT / PBEF ELISA Kit (MBL).

[0039] Next, after the skin model had been cultured, it was washed with PBS (-) and then divided into two equal parts. One half was used to measure the amount of melanin in the skin model, and the other half was used to measure the amount of NAD+ and NAMPT in the skin model.

[0040] The amount of melanin in the skin model was measured according to the following procedure. The collected skin model was extracted by infiltration overnight at 60°C using 0.1N NaOH, and then centrifuged to remove the film of the skin model and the insoluble fraction as a precipitate. The absorbance of the solution fraction was measured at 405 nm, and the amount of melanin was calculated based on the measurement results. The melanin inhibition rate of each group was calculated using the following formula, with the amount of melanin in the control group set at 100%.

number

[0041] The amount of NAMPT in the skin model was measured by the following procedure. The collected skin model was crushed and extracted at 4°C using 250μL of T-PER (trademark) Tissue Protein Extraction Reagent buffer (manufactured by Thermo Scientific) to prepare lysate. The crushing and extraction was carried out by adding the skin model, one zirconia ball, and T-PER Buffer to a polypropylene tube, and using a crushing extractor (bead-type cell crusher Micro Smash MS-100R (manufactured by Tommy Seiko Co., Ltd.)) at 4°C, 2000 rpm, and 5 minutes. The total protein concentration was calculated using a portion of the lysate (20μL) by relative comparison with BSA (bovine serum albumin) using the standard method of BCA protein assay (measured at OD562nm). The amount of NAMPT protein contained in 100μL of lysate was measured using CircuLex Human NAMPT / PBEF ELISA Kit (MBL Co., Ltd.). The measurement results were corrected to values ​​per unit protein amount.

[0042] The amount of NAD+ in the skin model was measured by the following procedure. The collected lysate from the skin model was deproteinized by spin column treatment using 10kD Spin Columns (ab93349, Abcam), and the eluate of the low molecular weight fraction was collected. The amount of NAD+ contained in the collected low molecular weight fraction was measured using the NAD / NADH Assay Kit (Colorimetric) (ab65348, Abcam).

[0043] Statistical processing was performed according to the following procedure. Bonferroni multiple comparisons were used as a multiple comparison test to evaluate the melanin suppression rate, and Dunnett test was used to evaluate the amount of NAMPT and the amount of NAD+. In each case, it was judged whether or not there was a significant improvement compared to the control group. * was used when p<0.05, and ** when p<0.01. In addition, it was judged that there was a significant tendency when 0.05≦p<0.1, and the p value was indicated.

[0044] Based on the measurement results when the drug was applied alone, the theoretical values ​​when the drug was applied in combination were calculated, and the obtained values ​​were compared with the actual values ​​when the drug was applied in combination to determine whether there was a synergistic effect from the combination of drugs. First, β-NMN and longan nikko were evaluated in groups added at 300 μg / mL, 100 μg / mL, and 30 μg / mL (ascorbic acid was evaluated only at 100 μg / mL). After confirming the concentration dependence of the measurement results at the above three points, the measurement results were fitted to an approximation equation of a logarithmic function or a quadratic function, and the predicted values ​​(theoretical values) when β-NMN and longan nikko were added at 50 μg / mL each were calculated from the obtained approximation equation. In each test, the total value of the "theoretical value when 50μg / mL of β-NMN is added alone" and the "theoretical value when 50μg / mL of longan berry is added alone" was compared with the "actual value when 50μg / mL of β-NMN and 50μg / mL of longan berry are added together," and if the actual value was 10% or more better, it was evaluated that the synergistic effect of β-NMN and longan berry was exerted. For ascorbic acid, 1 / 2 of the measured value of 100μg / mL was set as the "theoretical value when 50μg / mL of ascorbic acid is added alone" and compared with the "actual value when 50μg / mL of ascorbic acid and 50μg / mL of β-NMN are added together."

[0045] The evaluation results of the secretion amount of NAMPT contained in the culture medium are shown in Figure 1 (comparison of the effect of the combined use of ascorbic acid and β-NMN) and Figure 2 (comparison of the effect of the combined use of β-NMN and longan berry). While the actual measured values ​​of ascorbic acid and β-NMN were lower than the theoretical values, the actual measured values ​​of β-NMN and longan berry were significantly higher than the theoretical values, thus demonstrating the synergistic effect of the combination of β-NMN and longan berry in promoting the secretion of NAMPT.

[0046] The evaluation results of the melanin suppression rate in the skin model are shown in Figure 3 (comparison of the effect of the combined use of ascorbic acid and β-NMN) and Figure 4 (comparison of the effect of the combined use of β-NMN and longan berry). The actual measured values ​​of both ascorbic acid and β-NMN, and β-NMN and longan berry, greatly exceeded the theoretical values, thus demonstrating the synergistic effect of the combination of ascorbic acid and β-NMN, and β-NMN and longan berry on the suppression of melanin.

[0047] The evaluation results of the amount of NAMPT in the skin model are shown in Figure 5 (comparison of the effect of the combined use of ascorbic acid and β-NMN) and Figure 6 (comparison of the effect of the combined use of β-NMN and longan berry). While the actual values ​​of ascorbic acid and β-NMN were lower than the theoretical values, the actual values ​​of β-NMN and longan berry were significantly higher than the theoretical values, thus indicating a synergistic effect of the combination of β-NMN and longan berry in promoting the production of NAMPT.

[0048] The evaluation results of the amount of NAD+ in the skin model are shown in Figure 7 (comparison of the effect of the combined use of ascorbic acid and β-NMN) and Figure 8 (comparison of the effect of the combined use of β-NMN and longan berry). While the actual values ​​of ascorbic acid and β-NMN were lower than the theoretical values, the actual values ​​of β-NMN and longan berry were significantly higher than the theoretical values, thus indicating a synergistic effect of the combination of β-NMN and longan berry in promoting the production of NAD+.

Claims

1. A composition comprising NMN for use in combination with longan.

2. A composition comprising longan for use in combination with NMN.

3. A composition comprising NMN and longan.

4. The composition according to any one of claims 1 to 3 for skin whitening.

5. The composition according to any one of claims 1 to 3, for promoting NAMPT production.

6. The composition according to any one of claims 1 to 3 for promoting production of nicotinamide adenine dinucleotide.

7. The composition according to any one of claims 1 to 3, wherein CA / CB is 0.2 to 20, where CA is the dry weight of NMN in the composition and CB is the dry weight of longan.

8. The composition according to any one of claims 1 to 3, which is a food composition.

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