Searchin activator

Abstract

To provide an excellent sirtuin activator.SOLUTION: An improved sirtuin activator can be provided by combining Kaempferia parviflora, a Kaempferia parviflora extract, or a polyalkoxyflavonoid compound with nicotinamide mononucleotide (NMN).SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a sirtuin activator.

Background Art

[0002] Due to the recent development of medical technology and the like, the average life expectancy of humans has increased, and it has become an era called the "100-year life era." On the other hand, in Japan, the declining birthrate and aging population are progressing, and it is said that the proportion of the elderly aged 65 or older will reach about 40% of the Japanese population by 2060. As a result, a decrease in the number of domestic workers is expected, and there is concern that it may have an adverse impact on social development (see Non-Patent Documents 1 to 3). That is, for future social development, it is an important issue to realize not only a long-lived society but also an extension of healthy life expectancy.

[0003] The average life expectancy refers to the period indicating how many years one can live from birth, and the healthy life expectancy refers to the period during which one can live without being restricted in daily life due to health problems. That is, the difference between the average life expectancy and the healthy life expectancy means the period during which there are restrictions in daily life, that is, the period of being unhealthy. Currently, the difference between the average life expectancy and the healthy life expectancy is 9.13 years for men and 12.68 years for women, and although it is decreasing year by year, there is still room for improvement.

[0004] As one means of extending the healthy life expectancy, prevention of age-related diseases is considered important (Non-Patent Document 2). And for the prevention of age-related diseases, activation of an enzyme called sirtuin (hereinafter referred to as "sirtuin enzyme") is one important means. This sirtuin enzyme is an NAD + -dependent deacetylase, and it has been reported that it is involved in suppression of neurodegeneration, improvement of glucose metabolism and lipid metabolism, anti-obesity, etc. through various transcription factors, and it is an important enzyme for aging suppression and metabolic control. Therefore, it is desired to provide an anti-aging material having an excellent sirtuin activation effect (Non-Patent Document 4).

[0005] By the way, black ginger (Kaempferia parviflora) is a plant of the genus Kaempferia in the Zingiberaceae family, and is also called black ginger, Thai ginseng, and krachai dum. The origin of black ginger is in Thailand and Laos in Southeast Asia, where it has been consumed daily since ancient times and is used for nourishing the body, enhancing energy, lowering blood sugar levels, recovering physical strength, and improving the cardiovascular and digestive systems. So far, it has been described in Non-Patent Document 5 and Patent Documents 1 and 2 that the black ginger extract has the function of activating sirtuin.

[0006] Black ginger is known to contain five characteristic component polymethoxyflavonoid compounds (apigenin-5,7,4’-trimethylether, quercetin-3,5,7,3’,4’-pentamethylether, chrysin dimethylether, 3,5,7,4’-tetramethoxyflavone, 3,5,7-trimethoxyflavone). Also, it has been reported that all of these five polymethoxyflavonoid compounds have sirtuin activation effects (see, for example, Non-Patent Documents 5 and 6 below).

[0007] NMN is a precursor of nicotinamide adenine dinucleotide (hereinafter referred to as "NAD+") and is also used to generate NAD+. There are reports that NAD+ is an important electron carrier present in all species and is involved in the sirtuin enzyme reaction. Also, it is described in Non-Patent Document 7 below that the production ability of NMN decreases with aging and NAD+ decreases with age.

Prior Art Documents

Patent Documents

[0008]

Patent Document 1

[0009] [Non-Patent Document 1] Healthcare Management Research Unit, Center for Policy Vision Research, The University of Tokyo. Report on the Results of the Project for Formulating and Utilizing Healthcare Management Evaluation Indicators in the Project for Promoting the Creation of Industries for Extending Healthy Life Expectancy in Fiscal 2015 [Non-Patent Document 2] Ministry of Health, Labour and Welfare. Report on the National Health and Nutrition Survey in 2015, Results of the Lifestyle Habits Survey, Part 3 [Non-Patent Document 3] Materials of the 11th Promotion Expert Committee for Health Japan 21 (Second) [Non-Patent Document 4] Rahman, S., & Islam, R. (2011). “Mammalian Sirt1: insights on its biological functions.” Cell Communication and Signaling, 9, 11. [Non-Patent Document 5] Nakata, A., Koike, Y., Matsui, H., Shimada, T., Aburada, M., & Yang, J. (2014). “Potent SIRT1 enzyme-stimulating and anti-glycation activities of polymethoxyflavonoids from Kaempferia parviflora.” Natural product communications, 9(9), 1291 - 1294. [Non-Patent Document 6] Zhang, M., Lu, P., Terada, T., Sui, M., Furuta, H., Iida, K., Katayama, Y., Lu, Y., Okamoto, K., Suzuki, M., Asakura, T., Shimizu, K., Hakuno, F., Takahashi, S., Shimada, N., Yang, J., Ishikawa, T., Tatsuzaki, J., Nagata, K. (2021). “Quercetin 3,5,7,3‘,4’,-pentamethyl ether from Kaempferia parviflora directly and effectively activates human SIRT1.” Communication Biology, 4, 209. [Non-Patent Document 7] Bonkowski, M., Sinclair, D. (2017). “Slowing ageing by design: The rise of NAD+ and sirtuin-activating compounds.” Nature Reviews Molecular Cell Biology, 17(11), 679-690. [Summary of the Invention] [Problems to be Solved by the Invention]

[0010] As described above, with the deepening of the aging society, it is desired to provide a composition having a more excellent sirtuin activation effect.

[0011] However, the techniques described in Patent Documents 1 and 2 and Non-Patent Documents 5 to 7 are all only reports using single plant extracts or plant-derived components, and there is no report on the case of combining them.

[0012] Therefore, in view of the above problems, an object of the present invention is to provide a composition having a more excellent sirtuin activation effect by combining materials. [Means for Solving the Problems]

[0013] The inventors of the present invention have conducted intensive studies on the above problems. As a result, they have found that by combining black turmeric or a black turmeric extract or a polyalkoxylated flavonoid compound with NMN, specifically by simultaneously containing NMN, a better sirtuin activation effect than when used alone can be obtained, and thus the present invention has been completed.

[0014] That is, the sirtuin activator according to one aspect of the present invention is characterized by containing at least one of black turmeric and an extract of black turmeric (Kaempferia parviflora) and nicotinamide mononucleotide (NMN).

[0015] In addition, the sirtuin activator according to another aspect of the present invention is characterized by containing a polyalkoxylated flavonoid compound represented by the following general formula (I) and nicotinamide mononucleotide (NMN).

Chemical formula

[0016] In addition, in this aspect, although not limited, the polyalkoxylated flavonoid compound is preferably a polymethoxylated flavonoid compound represented by the following general formula (II).

Chemical formula

[0017] Also, from this perspective, it is preferably something containing at least one of black ginger (Kaempferia parviflora) and a black ginger extract.

[0018] Also, from this perspective, although not limited, the compound represented by the above general formula (I) or (II) preferably contains quercetin-3,5,7,3’,4’-pentamethyl ether.

[0019] Also, a food or drink, health food, cosmetic, or pharmaceutical composition according to another aspect of the present invention contains the sirtuin activator described in any of the above.

Advantages of the Invention

[0020] As described above, the present invention can provide a composition having a more excellent sirtuin activation effect.

Brief Description of the Drawings

[0021]

Figure 1

Figure 2

Modes for Carrying Out the Invention

[0022] Hereinafter, embodiments of the present invention will be described. However, the present invention can be implemented in many different forms and is not limited only to the specific examples shown in the following embodiments and examples.

[0023] One embodiment of the present invention is a composition having a more excellent sirtuin activation effect. That is, it is characterized by containing an extract of black turmeric and NMN, or containing quercetin-3,5,7,3’,4’-pentamethyl ether and NMN. With these combinations, a very excellent sirtuin activation effect is expected.

[0024] In this embodiment, the black turmeric used can be the rhizome of Kaempferia parviflora, which is native to Southeast Asia such as Thailand and Laos, or cultivated in Okinawa. Also, fresh, dried or processed rhizomes of black turmeric can be used, and the underground part of black turmeric including the rhizome can be used as it is.

[0025] Also, the extraction method of the black turmeric extract is not particularly limited and can be carried out according to methods well known to those skilled in the art. As the extraction solvent, water or warm water, alcohol-based solvents, and other organic solvents such as acetone can be used. Examples of alcohol-based solvents include methanol, ethanol, propanol, isopropanol, butanol, isobutanol, etc. Examples of other organic solvents other than acetone include esters such as ethyl acetate; polyhydric alcohols such as ethylene glycol, propylene glycol, 1,3-butylene glycol; ethers such as diethyl ether, etc. These solvents can be used alone or in combination. The extraction solvent may be used in the form of a water-containing hydrophilic solvent by combining water and a hydrophilic organic solvent. When using a combination of solvents, the mixing ratio of the solvents can be set arbitrarily. Among these, it is preferable to use ethanol, and it is more preferable to use water-containing ethanol with an ethanol content of 30% to 95% (v / v).

[0026] The amount of the extraction solvent is preferably 2 to 100 parts by weight based on the dry weight of the black turmeric rhizome. The extraction temperature is preferably 4 to 90°C. The extraction time is preferably 30 minutes to 1 week. The extraction method can be carried out by any method such as stirring extraction, immersion extraction, countercurrent extraction, ultrasonic extraction, supercritical extraction, etc.

[0027] The black turmeric extract includes any of the filtrate obtained by filtering the obtained extract, or the concentrated solution obtained by concentrating the filtrate, the dried product obtained by drying the concentrated solution, or the crude or purified product thereof. The concentration method can be carried out by any method such as evaporation concentration or membrane concentration. The drying method can be carried out by any method such as vacuum drying, freeze drying, spray drying, etc. If necessary, excipients such as dextrin may be added. When purification is carried out, it can be carried out according to means known to those skilled in the art. For example, synthetic adsorption resins, activated carbon, ion exchange resins, gel filtration agents such as Sephadex and Biogel, column chromatography, recrystallization, etc. may be used alone or in combination.

[0028]

Chemical formula

[0029] In the above formula, R1 to R7 each independently represent a lower alkoxy group having 1 to 6 carbon atoms, a hydrogen atom or a hydroxyl group.

[0030] The lower alkoxy group having 1 to 6 carbon atoms in R1 to R7 in the general formula (I) refers to a group in which an alkyl group having 1 to 6 carbon atoms is ether-bonded to an oxygen atom. Examples of the lower alkoxy group having 1 to 6 carbon atoms include a methoxy group, an ethoxy group, a propoxy group, a butoxy group, a pentoxy group, a hexoxy group, or structural isomers thereof, and preferably a methoxy group. Preferred polyalkoxyflavonoid compounds are polymethoxyflavonoid compounds, and more preferred polymethoxyflavonoid compounds are compounds represented by the following general formula (II).

Chemical formula

[0031] Also, R 11 ~R 16 each independently represent a hydrogen atom, a hydroxyl group or methoxy, and R13 represents a hydrogen atom or a methyl group.

[0032] Specific examples of the compound represented by the general formula (II) are shown in Table 1 below. [Table 1]

[0033] The polyalkoxylated flavonoid compounds represented by the general formula (I) or (II) can be obtained by extraction and purification from black ginger or citrus fruits in addition to chemical synthesis. The synthesis method is not particularly limited, and conventionally known methods can be used. Further, the polyalkoxylated flavonoid compounds represented by the general formula (I) or (II) can also be combined with at least one of black ginger and black ginger extracts.

[0034] The citrus fruits can be selected from the group consisting of Citrus depressa, C. tachibana, C. leiocarpa, C. tardiva, C. succosa, shikainkan, C. kinokuni, C. erythrosa, C. sunki, C. deliciosa, C. nobilis, C. retuculata, C. tangerina belonging to the mandarin group, and C. hanayu, C. nippokoreana belonging to the yuzu group.

[0035] Also, as a method for purifying the polyalkoxylated flavonoid compounds, for example, synthetic adsorption resins, activated carbon, ion exchange resins, gel filtration agents such as Sephadex and Biogel, column chromatography, recrystallization, etc. may be used alone or in combination.

[0036] The structure of the polyalkoxylated flavonoid compounds obtained by purification can be identified according to means known to those skilled in the art. For example, proton nuclear magnetic resonance spectroscopy (1 H-NMR), carbon nuclear magnetic resonance spectroscopy ( 13 C-NMR), mass spectrometry (MS), elemental analysis, infrared spectroscopy (IR), ultraviolet spectroscopy (UV), melting point measurement, etc. may be used alone or in combination.

[0037] The present turmeric extract preferably contains a total amount of 5 types of polymethoxyflavonoid compounds (apigenin-5,7,4'-trimethyl ether, quercetin-3,5,7,3',4'-pentamethyl ether, chrysin dimethyl ether, 3,5,7,4'-tetramethoxyflavone, 3,5,7-trimethoxyflavone) of 4% by weight or more, more preferably in the range of 10% by weight or more and 50% by weight or less. By being contained within this range, the effects of the present composition are preferably exhibited.

[0038] In the present composition, the turmeric extract and NMN are not particularly limited, but when the weight of NMN is 1, the amount of the turmeric extract is preferably in the range of 0.01 or more and 20 or less, more preferably 0.05 or more and 10 or less, and still more preferably 0.1 or more and 5 or less.

[0039] Also, polyalkoxyflavonoid and NMN are not particularly limited, but when the weight of NMN is 1, the amount of polyalkoxyflavonoid is preferably in the range of 0.0003 or more and 0.6 or less, more preferably 0.0015 or more and 0.3 or less, and still more preferably 0.003 or more and 0.15 or less.

[0040] Also, the present composition can take the form of pharmaceuticals or foods, but each extract is individually used for food and has been confirmed not to cause great harm to users. Therefore, if it is a food, it can be easily obtained without a doctor's prescription, and can particularly take the form of a food with functional claims or a health food.

[0041] Furthermore, the form of the present composition is not limited to solids and can also be in the form of beverages. When the present composition is a food, examples of its form include desserts such as drinks, candies, jellies, gummies, etc. When it is a health food or a food with functional claims, examples of its form include, in addition to the above, forms such as tablets, hard capsules, soft capsules, granules, drinks, etc.

[0042] As described above, by combining at least any one of black ginger (Kaempferia parviflora), black ginger extract, and polyalkoxylated flavonoid compounds with NMN, an excellent sirtuin activator can be provided.

Examples

[0043] Here, actually, the composition according to the above-described embodiment was manufactured and its effects were confirmed. The following is a specific description, but the present invention is not limited to the following examples.

[0044] (Black ginger extract) Dried chips of the rhizomes of black ginger (Kaempferia parviflora) were pulverized by a mixer, 80% ethanol was added to the pulverized material, and after heating under reflux and filtration, an extract was obtained. The extraction residue was again heated under reflux with 80% ethanol and then filtered to obtain an extract. The first extract and the second extract were combined, concentrated under reduced pressure, and then an excipient was added and dried under reduced pressure to obtain a black ginger extract (80% ethanol extract).

[0045] (Quercetin-3,5,7,3’,4’-pentamethyl ether) The 80% ethanol extract of black ginger obtained above was partitioned with water and ethyl acetate, concentrated and dried, and 4.58 g of an ethyl acetate phase was obtained. The obtained ethyl acetate phase was adsorbed onto silica gel and then eluted with a methanol-chloroform mixed solvent, and the obtained fraction was further isolated and purified by reverse-phase high-performance liquid chromatography (ODS) to obtain Compound 1 (784 mg).

[0046] Then, the nuclear magnetic resonance spectrum of the obtained Compound 1 was acquired. Table 2 below shows the results of proton nuclear magnetic resonance spectroscopy ( 1 1H-NMR) and carbon nuclear magnetic resonance spectroscopy ( 13 13C-NMR). Note that the "measured value" in the table indicates the result measured by the inventors, and the "literature value" indicates the result described in "Simultaneous identification and quantitation of 11 flavonoid constituents in Kaempferia parviflora by gas chromatography. Journal of Chromatography A, 1143:227 - 233, 2007."

Table 2

[0047] In addition, Table 3 below shows the results of the physical property data of Compound 1 measured using the melting point measurement method, infrared spectroscopy (IR), ultraviolet spectroscopy (UV), and mass spectrometry (MS).

Table 3

[0048] From the chemical shift values, spectral shapes, coupling constants, coupling states, comparison with literature values, and physical property data such as the melting point in the nuclear magnetic resonance spectrum of Compound 1 ( 1 1H-NMR and 13 13C-NMR), Compound 1 was identified as quercetin-3,5,7,3’,4’-pentamethylether.

[0049] (Evaluation Test of Sirtuin Activation Effect) 2.5×10 5Monocyte macrophage cells (RAW264.7) derived from mouse ascites at a density of cells / mL (Dulbecco's MEM containing 10% fetal bovine serum (FBS)) were seeded in a cell culture dish and cultured overnight at 37°C in a 5% CO₂ incubator. After the culture, the medium was replaced with fresh medium and cultured for an additional 1 hour. Then, 100 μL of NMN at 25, 50, or 100 μg / mL was added and left standing at 37°C in a 5% CO₂ incubator for 6 hours. After that, the NMN-containing medium was removed, 200 μL of cell lysis buffer (50 mM HEPES buffer pH 8, 50 mM NaCl, 1% Triton X-100) was added, and the cells were collected using a cell scraper. The collected cells were disrupted with a homogenizer, and the disrupted cell lysate was centrifuged at 12,000 rpm, and three kinds of supernatants were collected.

[0050] The black fungus extract or quercetin-3,5,7,3’,4’-pentamethyl ether was added to each of the three kinds of cell supernatants obtained above, and the sirtuin activation effect was evaluated using the FLUOR DE LYS (registered trademark) SIRT activity assay kit. Also, the final concentration of the black fungus extract was adjusted to 50 μg / mL, and the final concentration of quercetin-3,5,7,3’,4’-pentamethyl ether was adjusted to 2 μg / mL.

[0051] (Test Result 1) As shown in Figure 1, when the black fungus extract and NMN were added simultaneously, a stronger sirtuin activation effect was shown than that of the black fungus extract or NMN alone. Also, similar sirtuin activation effects were confirmed at ratios of 1:2, 1:1, and 2:1 of the black fungus extract to NMN. That is, according to this result, it was confirmed that a synergistic sirtuin activation effect superior to that of the black fungus extract alone and NMN alone can be obtained when the black fungus extract and NMN are added simultaneously.

[0052] (Test Result 2) As shown in Fig. 2, when quercetin-3,5,7,3’,4’-pentamethyl ether and NMN were added simultaneously, a stronger activating effect on sirtuin was shown than when each was added alone. That is, according to these results, it was confirmed that an excellent synergistic sirtuin-activating effect can be obtained when quercetin-3,5,7,3’,4’-pentamethyl ether and NMN are added simultaneously, compared with the case of quercetin-3,5,7,3’,4’-pentamethyl ether alone and the case of NMN alone.

[0053] As described above, it was confirmed that a more excellent sirtuin-activating effect can be obtained by combining the black ginger extract or quercetin-3,5,7,3’,4’-pentamethyl ether with NMN.

Industrial Applicability

[0054] By combining black ginger (Kaempferia parviflora), black ginger extract, or polyalkoxylated flavonoid compounds with nicotinamide mononucleotide (NMN), a more excellent sirtuin activator can be provided.

Claims

1. An activator for sirtuin (excluding those containing giant salamander collagen peptide), characterized by containing at least one of black ginger (Kaempferia parviflora) and an extract of black ginger, and nicotinamide mononucleotide (NMN).

2. The sirtuin activator according to Claim 1, characterized by containing at least one of the above-mentioned black ginger and the extract of black ginger containing a polyalkoxylated flavonoid compound represented by the following general formula (I), and the nicotinamide mononucleotide (NMN). 【Chemical Formula 1】 (R1 to R7 in the formula each independently represent a hydrogen atom, a hydroxyl group, or a lower alkoxy group having 1 to 6 carbon atoms).

3. The sirtuin activator according to Claim 2, wherein the polyalkoxylated flavonoid compound is a polymethoxylated flavonoid compound represented by the following general formula (II). 【Chemical Formula 2】 (R11, R12, R14, R15 and R16 in the formula each independently represent a hydrogen atom, a hydroxyl group, or methoxy, and R13 represents a hydrogen atom or a methyl group).

4. The sirtuin activator according to Claim 2 or 3, wherein the compound represented by the general formula (I) or (II) contains quercetin-3,5,7,3',4'-pentamethyl ether.

5. A food or drink, health food, cosmetic, or pharmaceutical composition, characterized by containing the sirtuin activator according to any one of Claims 1 to 4.

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