NAD production promoter

Abstract

To provide a composition that promotes the recovery of nicotinamide adenine dinucleotide (NAD), which decreases in a living body with aging.SOLUTION: The present invention discloses an NAD production promoter containing placenta extract and collagen peptides, and external preparations for skin, cosmetics, internal agents, food and drink compositions, and quasi drugs containing the NAD production promoter.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an NAD production promoter, particularly to an NAD production promoter that is highly safe and can be used both as an external preparation and as an internal preparation. [Background technology]

[0002] Nicotinamide adenine dinucleotide (NAD) is known to function as a coenzyme for dehydrogenase in the body. NAD is particularly involved in the glycolysis and citric acid cycle in the body, and greatly influences energy production.

[0003] It is known that the activity of nicotinamide phosphoribosyltransferase (NAMPT), an enzyme essential for maintaining intracellular NAD levels, decreases with age (see, for example, Non-Patent Document 1), and that the amount of NAD in the skin also decreases with age (see, for example, Non-Patent Document 2).

[0004] On the other hand, it is known that NAD intermediate metabolic products such as nicotinamide riboside (hereinafter referred to as NR) improve the pathology of aging-related diseases (see, for example, Non-Patent Document 3), and it is also known that NR is found in large amounts in milk (see, for example, Non-Patent Document 4). [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] PLos ONE.Vol.12,e0170930(2017) [Non-Patent Document 2] PLos ONE.Vol.7,e42357(2012) [Non-Patent Document 3] Biochemistry Vol. 87, No. 2, pp. 239-244 (2015) [Non-Patent Document 4] J.Nutr.Vol.146,pp.957-63(2016) Summary of the Invention [Problem to be solved by the invention]

[0006] It is desirable to develop a substance that can promote NAD production, particularly by promoting the recovery of NAD production in aging individuals, thereby preventing aging. [Means for solving the problem]

[0007] The present inventors discovered that the combined application of placenta extract and collagen peptide to a living body can promote the recovery of intracellular NAD, and thus completed the present invention.

[0008] That is, according to one embodiment, the present invention provides an NAD production promoter comprising a placenta extract and a collagen peptide.

[0009] In the NAD production promoter, the collagen peptide preferably contains a dipeptide or tripeptide containing Hyp.

[0010] In the NAD production promoter, the collagen peptide preferably contains Ala-Hyp.

[0011] In the NAD production promoter, the placenta extract preferably contains 6 ppm or more of NR per dry weight.

[0012] In the NAD production promoter, the weight ratio of the placenta extract to the collagen peptide is preferably 2000:1 to 10:1.

[0013] According to another embodiment, the present invention relates to an external skin preparation, a cosmetic, an internal preparation, a food and beverage composition, or a quasi-drug, which contains the NAD production promoter described in any one of the above. Effect of the Invention

[0014] According to the present invention, it is possible to provide an NAD production promoter that can significantly promote intracellular NAD production. The NAD production promoter can be used as an external preparation for the skin and an internal preparation, and can be very useful for preventing aging of the body. [Brief description of the drawings]

[0015] [Figure 1] FIG. 1 is a graph showing the relative content of NR in milk and placenta extract. [Diagram 2] FIG. 2 is a graph showing the relative amount of NAD production when the concentration of placenta extract was changed and the concentration of collagen peptide was kept constant and an NAD production promoter was added to three-dimensionally cultured skin. [Diagram 3] FIG. 3 is a graph showing the relative amount of NAD production when collagen peptide alone, placenta extract alone, and an NAD production promoter were added to a cell culture medium. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] Hereinafter, an embodiment of the present invention will be described with reference to the drawings. However, the present invention is not limited to the embodiment described below.

[0017] [First embodiment: NAD production promoter] According to a first embodiment, the present invention relates to an NAD production promoter, which contains a placenta extract and a collagen peptide.

[0018] In this embodiment, the placenta extract refers to an extract derived from the placenta of a mammal. The mammal includes, but is not limited to, humans, cows, pigs, sheep, and horses. Preferably, the placenta extract is derived from the placenta of a pig. The placenta extract may be one that has been subjected to heat treatment, or may be a placenta extract that is not heated and is called "raw placenta".

[0019] It is preferable to use raw placenta that has been prepared so as not to damage useful active ingredients such as cytokines contained in the placenta (e.g., TGFβ1, EGF, FGF1, IGF) as much as possible. This "raw placenta" can be prepared, for example, as follows. The collected placenta is trimmed to remove unnecessary tissue fragments and blood, and then sterilized using a method such as low-temperature sterilization and frozen storage. Thereafter, the placenta extract is extracted by thawing (i.e., by freezing and thawing), and then solid-liquid separation is performed by centrifugation. The supernatant obtained by solid-liquid separation is called placenta extract stock. Raw placenta can be obtained by removing viruses and bacteria from this placenta extract stock. As a method for removing viruses and bacteria, it is preferable to use a method that does not use acid or heat, such as a filtration process using multiple filtration membranes. The raw placenta obtained in this way has the advantage that useful active ingredients such as cytokines remain in large amounts because no sterilization method using heat or acid is used in the preparation stage.

[0020] The placenta extract is preferably one obtained by removing high molecular weight proteins and their aggregates from the raw placenta. The high molecular weight proteins referred to here are proteins with a molecular weight of approximately more than 100 kilodaltons. This makes it difficult for the components to aggregate due to denatured high molecular weight proteins, thereby suppressing the denaturation and deterioration of NR and increasing the content of NR. In addition, the content of low molecular weight proteins and free amino acids can be increased, thereby increasing the absorption efficiency into the body. The low molecular weight proteins are proteins with a molecular weight of approximately 20 kilodaltons or less. In particular, those with a total nitrogen content of 0.01 to 0.1% when quantified are preferred.

[0021] As a method for removing high molecular weight protein and its aggregates, for example, filter filtration separation can be used.Specifically, the raw placenta can be subjected to molecular weight fractionation treatment using a filter membrane.As the filter membrane, for example, a microfiltration membrane with a pore size of 0.22 μm or less, or an ultrafiltration membrane with a molecular weight cutoff of 20 kilodaltons or less can be used.

[0022] By further purifying the raw placenta by this method, the blending ratio of water-soluble vitamins such as niacin and NAD intermediate metabolites can be increased. The NAD intermediate metabolites referred to here are NR, nicotinamide mononucleotide, nicotinic acid riboside, nicotinic acid mononucleotide, and nicotinic acid adenine dinucleotide. It is preferable that such placenta extract contains a large amount of NR, and it is most preferable that the NR amount per dry weight is 6 ppm or more. By making the NR amount in the placenta extract 6 ppm or more, it is possible to significantly promote the production of NAD in cells when it is taken orally or externally. The upper limit of the NR amount is not particularly limited in theory, but for example, the NR amount may be 6 ppm or more and about 1200 ppm or less.

[0023] In addition to the above, commercially available placenta extracts can also be used. Commercially available placenta extracts include those sold as placenta extracts, placenta extract powders, placenta bulk powders, etc., and are used as raw materials for pharmaceuticals and supplements.

[0024] The collagen protein contained in the NAD production promoter according to this embodiment may be a hydrolysate of collagen derived from a living organism.

[0025] More specifically, collagen protein extracted from connective tissues such as skin, bone, and tendon of mammals and fish such as cows and pigs, or collagen protein obtained by hydrolysis or the like from gelatin, which is a thermally denatured product of collagen protein, may be used, or artificially synthesized collagen protein may be used. When obtained from collagen protein, a collagen protein solution is applied to a collagenase enzyme immobilized column and enzymatic decomposition is performed by the column method. After passing through the column, the solution after the enzyme reaction is separated and filtered through a 0.22 to 0.45 μm filter, and the filtrate is freeze-dried to powder to obtain collagen tripeptide. The collagen tripeptide powder is then redissolved and each dipeptide- and tripeptide-containing fraction is purified by HPLC (gel filtration and ODS column). Furthermore, in order to obtain a single collagen tripeptide component, it can be purified using ion exchange chromatography and a carbon column. When obtained by artificial synthesis, artificial synthesis is performed by solid phase method using a peptide synthesis device. The peptide chain is extended from the C-terminus by the Fmoc method according to the program of the device used. To prevent side reactions, a carrier that is a support for solid-phase peptide synthesis is used, and an Fmoc amino derivative of ornithine in which the α-amino group and δ-amino group are protected with Fmoc and Boc in advance and β-alanine in which the β-amino group is protected with Fmoc is used. Fmoc-Orn(Boc) is bound to the carrier, the α-amino group is deprotected with piperidine, etc., and then the C-terminus of Fmoc-Orn(Boc) is coupled, the α-amino group of the second ornithine is deprotected with piperidine, etc., and the C-terminus of Fmoc-β-Ala is similarly coupled. Finally, all deprotection and carrier removal are performed with an acid such as trifluoroacetic acid, and the dipeptide and tripeptide are extracted with 0.01-2% trifluoroacetic acid water and lyophilized to produce the dipeptide and tripeptide, which can be obtained by purifying with HPLC (reverse flow column) or the like.

[0026] 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 more mixtures selected from dipeptides or tripeptides of Ala-Hyp-Gly, Pro-Hyp-Gly, Pro-Hyp, Leu-Hyp, Ala-Hyp. Among them, it is most preferable to contain the dipeptide of Ala-Hyp. Since Ala-Hyp has a lower molecular weight than tripeptides, it is preferably absorbed well into the body and has high stability among these peptides.

[0027] The placenta extract and the collagen peptide are preferably contained such that the weight ratio of the placenta extract to the collagen peptide is 2000:1 to 10:1, and more preferably 1000:1 to 100:1. By containing the placenta extract and the collagen peptide within the above ratio range, the collagen peptide can contribute to the stability of NR in the placenta extract and increase the production amount of NAD.

[0028] In the NAD production promoter according to this embodiment, by containing both the placenta extract and the collagen peptide, the production of NAD can be promoted. More specifically, it is considered that by suppressing the degradation of NR contained in the placenta extract by the collagen peptide, a decrease in the amount of NR, which is an intermediate metabolite of NAD, can be suppressed, and as a result, the amount of NAD produced in the living body can be promoted. In addition, the NAD production promoter also has a moisturizing effect, a whitening effect, an anti-wrinkle effect, an anti-inflammatory effect, a tyrosinase inhibitory effect, a wound healing promoting effect, a fibroblast proliferation promoting effect, a nerve cell activation effect, a blood pressure increase suppressing effect, an elastase inhibitory effect, which are the actions of the conventionally known placenta extract alone, and a promoting effect on the synthesis of in vivo collagen, which is the action of the collagen peptide alone.

[0029] In addition to placenta extract and collagen peptides, the NAD production promoter may include optional ingredients such as nicotinamide, vitamin C derivatives, arbutin, ellagic acid, kojic acid, tranexamic acid, glycerin, 1,3-butylene glycol, and propylene glycol.

[0030] The NAD production promoter can be obtained by mixing a placenta extract and a collagen peptide. The obtained NAD production promoter is a liquid composition containing a placenta extract and a collagen peptide. The NAD production promoter can be stably stored for about 1 to 3 years in a refrigerator 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 have reduced NAD production, so it can also be called an NAD recovery promoter.

[0031] The NAD production promoter can be added to internal medicines, food and drink compositions, skin care products, cosmetics, and quasi-drugs. In particular, it can be added to food and drink, cosmetics, etc., to obtain anti-aging effects. Various compositions to which the NAD production promoter is added are described below.

[0032] [Second embodiment: oral preparation] According to a second embodiment of the present invention, there is provided an internal medicine, which contains the NAD production promoter according to the first embodiment. The internal medicine may be, for example, an orally administered medicine, and examples of such medicine include tea, capsules, tablets, pills, granules, fine granules, syrup, dry syrup, and the like.

[0033] These oral preparations, including the NAD production promoter according to the first embodiment, are manufactured by conventional methods generally known in this field. The oral 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 oral preparations are used consisting of a liquid composition, they may be pharmacologically acceptable emulsions, solutions, suspensions, syrups, etc., and for example, purified water, ethanol, etc. are used as carriers. In addition, adjuvants such as wetting agents and suspensions, sweeteners, flavorings, preservatives, etc. may be added if desired.

[0034] The amount of the active ingredient of the NAD production promoter in the oral preparation is, for example, based on the dry weight of the placenta extract, preferably 50 mg or more, preferably 100 mg or more per day for an adult. The upper limit of the dosage is preferably 10,000 mg or less, more preferably 5,000 mg or less per day. Therefore, the NAD production promoter may be contained in the range according to each form so as to achieve the above-mentioned intake amount.

[0035] [Third embodiment: food and beverage composition] According to a third embodiment of the present invention, there is provided a food and drink composition, which contains the NAD production promoter according to the first embodiment. The food and drink composition according to this embodiment can contain the NAD production promoter according to the first embodiment together with any optional ingredient normally used in food and drink compositions. Examples of such food and drink compositions for humans or animals include breads, confectioneries, noodles, meat products, processed 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.

[0036] Furthermore, food and drink compositions include, for example, nutritional supplements such as vitamins, nutritional supplement drinks, health foods for animals, foods for specified health uses, foods with nutritional functions, foods with health functions, etc., and can be commercialized in various commonly used forms, for example, 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 NAD production promoter according to the first embodiment.

[0037] These food and drink compositions are produced by conventional methods generally known in the art.

[0038] The amount of the active ingredient of the NAD production promoter in such a food and drink composition may be added so as to obtain a desired intake amount based on the weight of the placenta extract and 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 0.1 to 20% by weight. For example, it is preferable to add it to liquid food and drink within the range of 0.1 to 20% by weight, to solid food and drink within the range of 0.1 to 20% by weight, and to tablet food and drink within the range of 0.1 to 20% by weight. More specifically, it is preferable to add it to supplements within the range of 0.1 to 20% by weight based on the dry weight of the placenta extract, but is not limited thereto. 。

[0039] [Fourth embodiment: skin topical preparation] According to a fourth embodiment, the present invention provides an external preparation for skin, which includes the NAD production promoter according to the first embodiment. More specifically, the external preparation for skin may be a cosmetic, and examples of such cosmetics include anti-aging cosmetics, cleansing cosmetics, hair cosmetics, bath cosmetics, and medical cosmetics, as well as skin care cosmetics such as pigments, lotions, beauty serums, milky lotions, creams, gels, packs, liposomes, liquid, clay, and solid powder cosmetics, aerosol cosmetics, and poultices, as well as make-up cosmetics such as base creams and foundations.

[0040] The cosmetic product according to the present embodiment may contain water, alcohol, surfactants (cationic, anionic, nonionic, amphoteric surfactants, etc.), moisturizers (glycerin, 1,3-butylene glycol, propylene glycol, amino acids, urea, pyrrolidone carboxylates, nucleic acids, monosaccharides, oligosaccharides, etc., and derivatives thereof, etc.), thickeners (polysaccharides, polyacrylates, carboxyvinyl polymers, polyvinylpyrrolidone, polyvinyl alcohol, chitin, chitosan, alginic acid, carrageenan, xanthan gum, methylcellulose, etc., and derivatives thereof, etc.), and / or other additives. Conductive agents, etc.), wax, petrolatum, saturated hydrocarbon fatty acids, unsaturated fatty acids, silicone oils and their derivatives, triglycerides such as tri(caprylic / capric acid)glyceryl, glyceryl trioctanoate, ester oils such as isopropyl stearate, natural oils and fats (olive oil, camellia oil, avocado oil, almond oil, cacao butter, evening primrose oil, grape seed oil, macadamia nut oil, eucalyptus oil, rosehip oil, squalane, orange roughy oil, lanolin, ceramide, etc.), preservatives (oxybenzoic acid derivatives, dehydroacetic acid salts, photosensitizers, sorbic acid, phenoxyethanol, etc. and their derivatives, etc.), disinfectants (sulfur, trichlorocarbanilide, salicylic acid, zinc pyrithione, hinokitiol, etc. and their derivatives, etc.), ultraviolet absorbers (para-aminobenzoic acid, methoxycinnamic acid, etc. and their derivatives, etc.), anti-inflammatory agents (allantoin, glycyrrhizic acid, etc. and their derivatives, etc.), antioxidants (tocopherol, BHA, BHT, etc. and their derivatives, etc.), chelating agents (edetic acid, hydroxyethanediphosphonic acid, etc. and and their derivatives), animal and plant extracts (Angelica angelica, aloe, angelica tree, Scutellaria baicalensis, Phellodendron bark, seaweed, Chinese quince, chamomile, licorice, kiwi, cucumber, mulberry, birch, angelica tree, garlic, peony, hops, horse chestnut, lavender, rosemary, eucalyptus, milk, various peptides, placenta, royal jelly, etc., and refined or fermented products containing these ingredients, etc.), pH adjusters (inorganic acids, inorganic salts, organic acids, organic salts, etc., and their derivatives, etc.), vitamins (vitamin A, vitamin B, vitamin C, vitamin D)In addition, the following may be blended within the scope of the present invention: titanium oxide, talc, mica, silica, zinc oxide, iron oxide, silicon, and powders obtained by processing these substances. Note that the components constituting the cosmetic product are not limited to those mentioned above, and any components that can be used in cosmetics may be freely selected.

[0041] In addition to the above ingredients, the poultice may contain bases (kaolin, bentonite, etc.) and gelling agents (polyacrylates, polyvinyl alcohol, etc.) within the range that achieves the object of the present invention.The bath agent may be prepared into powder or liquid types by appropriately blending sulfates, bicarbonates, borates, colorants, and moisturizing agents within the range that achieves the object of the present invention.

[0042] These cosmetic products are produced by conventional methods generally known in the art.

[0043] The amount of the active ingredient of the NAD production promoter in the cosmetic product may be added based on the weight of the placenta extract so as to achieve the aforementioned intake amount within the range according to the form of the cosmetic product, and it is preferable to add it to the cosmetic product within the range of 0.1 to 20% by weight. For example, it is preferable to add it to the cosmetic product within the range of 0.1 to 20% by weight for liquid cosmetics, 0.1 to 10% by weight for cream cosmetics, and 0.1 to 5% by weight for powder cosmetics. More specifically, it is not limited to the following, but it is preferable to add it to the cosmetic product within the range of 1 to 10% by weight for lotion and 1 to 5% by weight for cosmetic cream.

[0044] [Fifth embodiment: quasi-drug] According to a fifth embodiment of the present invention, there is provided a quasi-drug, which includes the NAD production promoter according to the first embodiment. The quasi-drug includes candies, gums, toothpastes, liquid dentifrices, gel dentifrices, antiperspirants, beverages, and the like.

[0045] Quasi-drugs containing an NAD production promoter are produced by conventional methods generally known in this field.

[0046] The amount of the active ingredient of the NAD production promoter in the quasi-drug may be added based on the weight of the placenta extract so as to be the intake amount described above within the range according to the form of the quasi-drug, and it is preferable to add it to the quasi-drug in the range of 0.1 to 20% by weight. For example, it is preferable to add it to a solid quasi-drug in the range of 0.1 to 20% by weight, a liquid quasi-drug in the range of 0.1 to 20% by weight, and a gel or paste quasi-drug in the range of 0.1 to 20% by weight. More specifically, it is preferable to add it to pet gum in the range of 0.02 to 5% by weight, and to toothpaste in the range of 0.02 to 1% by weight, although it is not limited thereto. EXAMPLES

[0047] 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.

[0048] <1: Measurement of NR content in placenta extract> NR in the sample solution was fluorescently derivatized and quantitatively analyzed by high performance liquid chromatography (HPLC). NR was derivatized according to the nicotinamide mononucleotide quantitative method (Formentini, L. et al., Biochem. Pharmacol. Vol. 77, pp. 1612-20(2009)) as follows.

[0049] The sample solutions used were placenta extract and milk. The placenta extract used was a stepwise filter-filtered porcine placenta extract. The stepwise filter-filter was first filtered using a microfiltration membrane with a pore size of 0.22 μm (manufactured by Sartorius), and then filtered using a centrifugal ultrafiltration membrane with a molecular weight cutoff of 10 kilodaltons (manufactured by Merck Millipore) to remove high molecular weight proteins and their aggregates. In addition, a commercially available milk product with a non-fat milk solid content of 8.3% or more and a milk fat content of 3.5% or more was used. 1N potassium hydroxide and 20% acetophenone were added to each sample solution, and the mixture was left to stand at 4°C for 15 minutes. Then, 88% formic acid was added, mixed well, and heated at 100°C for 5 minutes. This liquid was filtered through a membrane filter with a pore size of 0.45 μm to prepare a fluorescent derivatized sample solution. Separately, β-NR trifluoromethanesulfonate (Carbosynth) was dissolved in water at various concentrations and treated in the same manner as the sample solutions to prepare fluorescent derivatization standard solutions.

[0050] HPLC analysis was performed on the fluorescently derivatized sample solution and standard solution under the following conditions, a calibration curve was created using the peak area values ​​obtained from the standard solution, and the NR concentration was calculated from the peak area values ​​obtained from the sample solution. Figure 1 shows the relative values ​​to milk. Equipment: PU-2089Plus (JASCO) Detector: FP-2025Plus (JASCO) Excitation wavelength: 360 nm Fluorescence wavelength: 445nm Column: Inertsil ODS-3 (4.6 mm × 250 mm, 5 μm) (GL Science) Guard column: ODS-HG (4.0 mm x 10 mm, 5 μm) (Nomura Chemical) Flow rate: 1mL / min Mobile phase: Water (containing 0.05% trifluoroacetic acid):Acetonitrile (83:17)

[0051] As a result of the measurements, it was revealed that the placenta extract contains a higher concentration of NR than milk, which is known to be rich in NR (Non-Patent Document 4).

[0052] <2: Evaluation of NAD production promoter activity in human skin> The NAMPT inhibitor FK866 was added to the culture medium of 3D cultured skin (MatTek), and the NAD production-promoting activity of the NAD production promoter was measured in a state where NAD was depleted.

[0053] 5 nM FK866 was added to the culture medium of the 3D cultured skin, and NAD production promoter or PBS as a negative control was added to the upper side of the skin. 2 The cells were cultured at 37°C for 48 hours. The NAD production promoter was prepared so that the placenta extract was 1%, 5%, or 25% by volume in the culture medium, and the artificially synthesized dipeptide Ala-Hyp was 0.5 mM. The cultured skin cells were then eluted, and the total amount of NAD in the cells was measured using the NAD / NADH Assay Kit-WST (manufactured by DOJINDO). At the same time, the protein amount in the cell eluate was measured by the BCA method, and the amount of NAD relative to the amount of protein in each cell eluate was calculated. Figure 2 shows the relative values ​​to the control (PBS).

[0054] As shown in Figure 2, a concentration-dependent increase in intracellular NAD levels in 3D cultured skin was observed with placenta extract, suggesting that placenta extract has NAD production-promoting activity.

[0055] <3: Verification of synergistic effects of placenta extract and Ala-Hyp> We investigated the effect of pretreatment with Ala-Hyp (JP Patent Publication No. 2010-24200), a dipeptide that has been confirmed to promote collagen synthesis in human skin fibroblasts, on the intracellular NAD production promoting activity of placenta extract.

[0056] Normal human dermal fibroblasts (Kurabo) were placed in a 12-well plate at 5 × 10 4 Cells were seeded at 10 cells / well and incubated in 5% CO 2The cells were cultured at 37°C for 24 hours, and then replaced with DMEM / F12 medium (Thermo Fisher Scientific). Artificially synthesized Ala-Hyp was added to the cells at 0.5 mM, and after 24 hours of culture, the cells were replaced with DMEM / F12 medium containing 5 nM FK866, and placenta extract was added to the cells at 5% by volume. After further 24 hours of culture, the cells were eluted, and the total amount of NAD in the cells was measured using the NAD / NADH Assay Kit-WST. At the same time, the protein in the cell eluate was measured by the BCA method, and the amount of NAD relative to the protein in each cell eluate was calculated. Figure 3 shows the relative values ​​to untreated (control) cells.

[0057] In a monolayer culture test using fibroblasts, it was confirmed that the addition of placenta extract increased the amount of intracellular NAD, and the increase was further enhanced by pretreatment with Ala-Hyp.

[0058] <4: NR stability test> NR is an unstable component in aqueous solution and is known to easily decompose into nicotinamide and ribose (JP Patent Publication No. 2017-518306). Here, we examined the inhibitory effect of Ala-Hyp on the decomposition of NR.

[0059] β-NR trifluoromethanesulfonate was prepared to 200 μM to prepare the NR solution, and β-NR trifluoromethanesulfonate aqueous solution and Ala-Hyp aqueous solution were mixed at a ratio of 1:1 to prepare the NR+Ala-Hyp solution with final concentrations of 200 μM and 5 mM, respectively. Each solution was left at room temperature (RT), 50°C, and 75°C for 1 hour, and then filtered through a membrane filter with a pore size of 0.45 μm to prepare a sample solution. Separately, β-NR trifluoromethanesulfonate was dissolved in water at each concentration and filtered through a membrane filter with a pore size of 0.45 μm to prepare a standard solution. Each sample solution and standard solution were subjected to HPLC analysis under the following conditions, a calibration curve was created using the peak area value obtained from the standard solution, and the NR concentration was calculated from the peak area value obtained from the sample solution. Equipment: PU-2089Plus (JASCO) Detector: MD-2010Plus (JASCO) Measurement wavelength: 220nm Column: Inertsil ODS-3 (4.6 mm × 250 mm, 5 μm) (GL Science) Guard column: ODS-HG (4.0 mm x 10 mm, 5 μm) (Nomura Chemical) Flow rate: 1mL / min Mobile phase: Water (containing 0.05% trifluoroacetic acid)

[0060] As a result of the measurement, it was confirmed that the decomposition reaction of NR is promoted in a temperature-dependent manner. At that time, it was found that the addition of Ala-Hyp tended to suppress the decomposition of NR. The results are shown in Table 1 below. Table 1 suggests that Ala-Hyp may contribute to the stability of NR in aqueous solution.

[0061] [Table 1]

[0062] <Prescription Examples> [Table 2]

[0063] [Table 3]

[0064] [Table 4]

[0065] [Table 5]

Claims

1. Placenta extract, and a collagen peptide containing one or more dipeptides or tripeptides selected from Ala-Hyp-Gly, Pro-Hyp-Gly, Pro-Hyp, Leu-Hyp, and Ala-Hyp, An NAD production promoter, wherein the weight ratio of the placenta extract to the collagen peptide is 2000:1 to 10:

1.

2. The NAD production promoter according to claim 1 , wherein the collagen peptide comprises the dipeptide Ala-Hyp.

3. The NAD production promoter according to claim 1 or 2, wherein the placenta extract contains 6 ppm or more of NR per dry weight.

4. An external skin preparation comprising the NAD production promoter according to any one of claims 1 to 3.

5. A cosmetic comprising the NAD production promoter according to any one of claims 1 to 3.

6. An oral preparation comprising the NAD production promoter according to any one of claims 1 to 3.

7. A food or drink composition comprising the NAD production promoter according to any one of claims 1 to 3.

8. A quasi-drug comprising the NAD production promoter according to any one of claims 1 to 3.

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