JP2021187789A - Heat shock protein inducer, nitric oxide production promoter, anti-menopausal disorder agent, anti-aging agent, cosmetic preparation and food or beverage - Google Patents

Abstract

To provide a heat shock protein inducer having a high heat shock protein inducing ability.SOLUTION: A heat shock protein inducer contains at least one component of diosgenin, hirsutine and glycyrrhizin.SELECTED DRAWING: Figure 2

Description

The present invention relates to heat shock protein inducers, nitric oxide production promoters, anti-menopausal agents, anti-aging agents, cosmetics and foods or beverages.

NO (nitric oxide) activates guanylate cyclase when it diffuses into vascular smooth muscle cells surrounding vascular endothelial cells according to a concentration gradient. Activated guanylate cyclase promotes the reaction to produce cGMP from GTP. cGMP has a function of activating cGMP-dependent protein kinase (also called protein kinase G, G kinase, PKG), and the activated cGMP-dependent protein kinase ^{promotes the pumping of Ca 2+ to} the outside of the cell, etc. ^{By reducing the concentration of Ca 2+} in smooth muscle cells, it has the effect of relaxing vascular smooth muscle and dilating blood vessels.
In order to promote such an action, for example, a nitric oxide production promoter containing hirsutine as an active ingredient has been proposed (see, for example, Patent Document 1).

Further, in recent years, heat shock protein (Hsp), which is a protein that is expressed when cells are stressed by heat, ultraviolet rays, active oxygen, etc. and protects cells, has attracted attention, and a heat shock protein expression inducer has been proposed. (See, for example, Patent Document 2).

Japanese Unexamined Patent Publication No. 2018-158907 International Publication No. 2012/043808

Patent Document 2 relates to the induction of Hsp70 among various heat shock proteins, and is also required to induce other types of heat shock proteins, and further has an effect of promoting nitric oxide production in Patent Document 1. Further improvement is required.

An object of the present invention is to provide a heat shock protein inducer having a high heat shock protein inducing ability and a nitric oxide production promoter having a high nitric oxide production promoting ability, and these heat shock protein inducers or To provide an anti-menopausal disorder agent, an anti-aging agent, a cosmetic, a food or a beverage containing a nitric oxide production promoter.

As a result of diligent studies, the present inventors have found that the above object can be achieved by blending a predetermined substance with hirsutine, and have completed the present invention.

That is, according to the present invention, (1) a heat shock protein inducer containing at least one component of diosgenin, hirsutine and glycyrrhizin.
(2) The heat shock protein inducer according to (1), which contains three components of diosgenin, hirsutine and glycyrrhizin.
(3) The heat shock protein inducer according to claim 2, wherein the amount of diosgenin is 50% by weight or less, the amount of hirsutine is 50% by weight or less, and the amount of glycyrrhizin is 50% by weight or less.
(4) The heat shock protein inducer according to any one of (1) to (3), wherein the heat shock protein is at least one of gp96, Hsp32, Hsp47, Hsp70, and Hsp90.
(5) An anti-menopausal agent containing the heat shock protein inducer according to any one of (1) to (4).
(6) An anti-aging agent containing the heat shock protein inducer according to any one of (1) to (4).
(7) Cosmetics containing the heat shock protein inducer according to any one of (1) to (4).
(8) A food or beverage containing the heat shock protein inducer according to any one of (1) to (4).
(9) A nitric oxide production promoter containing at least one component of diosgenin or glycyrrhizin,
(10) The nitric oxide production promoter according to (9), which contains the three components of diosgenin, hirsutine and glycyrrhizin.
(11) The nitric oxide production promoter according to (10), wherein the amount of diosgenin is 50% by weight or less, the amount of hirsutine is 50% by weight or less, and the amount of glycyrrhizin is 50% by weight or less.
(12) An anti-menopausal agent containing the nitric oxide production promoter according to any one of (9) to (11).
(13) An anti-aging agent containing the nitric oxide production promoter according to any one of (9) to (11).
(14) A cosmetic product containing the nitric oxide production promoter according to any one of (9) to (11).
(15) A food or beverage containing the nitric oxide production promoter according to any one of (9) to (11).
Is provided.

According to the present invention, it is possible to provide a heat shock protein inducer having a high heat shock protein inducing ability and a nitric oxide production promoting agent having a high nitric oxide production promoting ability. In addition, anti-menopausal agents, anti-aging agents, cosmetics, foods or beverages containing the heat shock protein inducer or nitric oxide production promoter can be provided.

It is a graph which shows the nitric oxide production amount of the diosgenin, hirsutine, glycyrrhizin and the three-component compounded product in the human vascular smooth muscle cell obtained in the Example. It is a graph which shows the mRNA inducing ability of the nitric oxide synthase of diosgenin, hirsutine, glycyrrhizin and the tri-component combination in human vascular smooth muscle cells obtained in Example. It is a graph which shows the heat shock protein inducing ability (1 hour) of the diosgenin, hirsutine, glycyrrhizin and the three-component combination in the mouse nerve cell obtained in an Example. It is a graph which shows the heat shock protein inducing ability (4 hours) of the diosgenin, hirsutine, glycyrrhizin and the three-component combination in the mouse nerve cell obtained in an Example.

Hereinafter, the nitric oxide production promoter and the heat shock protein inducer of the present invention will be described. The nitric oxide production promoter and heat shock protein inducer of the present invention contain diosgenin, hirsutine and glycyrrhizin.

(Diosgenin)
Diosgenin is a kind of saponin (steroid saponin) having a steroid skeleton having a structure represented by the following structural formula (1).

Further, diosgenin may be a chemically synthesized product or may be derived from a plant. As a chemically synthesized product, a commercially available product can also be used. Further, the plant-derived plant is not particularly limited as a plant, but for example, a plant such as the genus Yam of the family Yam such as yam, dioscorea bulbifer, and yam, and the genus Trigonella of the subfamily Papilionoideae such as fenugreek. Can be extracted from. The extraction may be any part of the whole plant, for example fruits, flowers, seeds, leaves, branches, bark, stems, stems, thorns, or roots, but roots (especially lump roots). It is preferable to use it. In addition, raw plants may be used for extraction, or dried plants may be used. As a method for extracting glycyrrhizin from a plant, the same method as the method for extracting hirsutine from a plant, which will be described later, can be adopted.

Further, in the present invention, as the plant-derived diosgenin, those obtained as diosgenin glycosides may be used. Further, as long as the effect of the present invention is not impaired, a hydrogen atom, a hydroxyl group or the like in diosgenin may be substituted with a substituent, or a diosgenin derivative chemically modified with a modifying group may be used.

(Hirsutine)
Hirsutine is a kind of indole alkaloid based on the indole skeleton, and has a structure represented by the following structural formula (2).

Hirsutine may be a chemically synthesized product or may be derived from a plant. When hirsutine is derived from a plant, the type of plant is not particularly limited as long as it is a plant containing hirsutine, for example, Uncaria rhynchophylla (Miq.) Miq., Uncaria sinensis (Oliv.) Havil, Examples include Rhododendron anthopogoides, Eucommia ulmoides Oliv., Polygonum caspitatum, and Amsonia elliptica (Thunb.) Roem. Et Schult. Any part of the whole plant may be used for extraction, for example, fruits, flowers, seeds, leaves, branches, bark, stems, stems, thorns, or roots. In addition, raw plants may be used for extraction, or dried plants may be used.

The type of solvent used to extract hirstine from plants can be appropriately selected by those skilled in the art, for example, water, ethanol, methanol, acetone, ethyl acetate, glycerin, propylene glycol, 1,3-butylene glycol. , Dimethylsulfonate, acetonitrile, N, N-dimethylformamide, 2-propanol, 1,4-dioxane, hexane, chloroform, dichloromethane, or a mixed solvent of two or more solvents selected from these, water. , Ethanol, or a mixed solvent of water and ethanol is more preferable.

Water or a mixed solvent with water may contain a salt, and examples thereof include physiological saline and a buffer solution. The type of salt used for the buffer is not particularly limited, and examples thereof include citrate, malate, phosphate, acetate and carbonate.

The degree of extraction is not particularly limited, and it may be purified to pure hirsutine, or it may be crudely purified. In the present specification, those extracted from plants at any extraction stage leading to pure hirsutine are referred to as plant extracts. The plant extract may be an extract that does not remove the solvent, or may be made solid by removing the solvent. The method for removing the solvent from the extract is not particularly limited, and known methods such as vacuum distillation, freeze-drying, and spray-drying (spray drying) can be used. In the present invention, the hirsutine analog described in JP-A-2018-158907 may be used.

(Glycyrrhizin)
Glycyrrhizin is a triterpene glycoside having a structure represented by the following structural formula (3). Glycyrrhizin is also called glycyrrhizic acid.

Glycyrrhizin can be obtained from licorice as a raw material. As the licorice, for example, plants belonging to the genus Glycyrrhiza of the leguminous family such as Glycyrrhiza urarensis or Glycyrrhiza glabra and Glycyrrhiza inflata can be used. Any part of the whole plant may be used for extraction, for example, fruits, flowers, seeds, leaves, branches, bark, stems, stems, thorns, or roots. In addition, raw plants may be used for extraction, or dried plants may be used.
As a method for extracting glycyrrhizin from a plant, the same method as the above-mentioned method for extracting hirsutine from a plant can be adopted.

As the glycyrrhizin, a pharmacologically acceptable salt of glycyrrhizin can be used as long as the effect of the present invention is not impaired. Examples of such a salt of glycyrrhizin include alkali metal salts such as sodium salt, potassium salt and dipotassium salt, ammonium salt and the like.

(Combined product of three ingredients)
In the present invention, the compounding ratio of each component of the three-component compound containing diosgenin, hirsutine and glycyrrhizin is preferably 0 to 50%, 0 to 50%, 0 to 50%, preferably 1 to 50%, based on the weight. , 1-50%, 1-50%, more preferably 10-50%, 10-50%, 10-50%, particularly preferably 20-40%, 20-40%, 20-40%.

Here, for example, when an arbitrary substance is selected and used in combination with hirsutine, there are many combinations that inhibit each other's effects. The same applies to the case where an arbitrary substance is selected and used in combination with diosgenin, or the case where an arbitrary substance is selected and used in combination with glycyrrhizin. On the other hand, in the present invention, by blending the three components of diosgenin, hirsutine and glycyrrhizin, their effects are not inhibited from each other.

Further, in the present invention, it may be in a form containing at least one component of diosgenin, hirsutine and glycyrrhizin. That is, in addition to the three-component compounded product of diosgenin, hirsutine and glycyrrhizin, the one-component compounded product ((1) diosgenin one-component compounded product, (2) hirsutine one-component compounded product, (3) glycyrrhizin one-component compounded product), two-component compounded product. It may be a compounded product ((4) a two-component compounded product of diosgenin and hirsutine, (5) a two-component compounded product of diosgenin and glycyrrhizin, and (6) a two-component compounded product of hirsutine and glycyrrhizin).

Further, in the case of a two-component compounded product, the compounding ratio of the two components is based on the weight, and in any case, it is preferably 10:90 to 90:10, more preferably 20:80 to 80:20, and even more preferably. It is 30:70 to 70:30, most preferably 40:60 to 60:40.

(Use)
The three-component compound containing diosgenin, hirsutine and glycyrrhizin is excellent in nitric oxide producing ability and also excellent in heat shock protein inducing ability. Therefore, it has an effective effect on menopause and anti-aging regardless of gender. Further, the above-mentioned one-component compounded product and two-component compounded product have the same effect.

In addition, since the three-component combination product containing diosgenin, hirsutine and glycyrrhizin has an action effect on menopausal disorders and an anti-aging effect, it can be added to a drug for these purposes. As a pharmaceutical product, it can be used as either a preventive drug or a therapeutic drug. Further, the above-mentioned one-component compounded product and two-component compounded product can also be used for the same purpose.

When blended in a pharmaceutical product, the three-component compound may be used alone, or it may be mixed with an additive generally acceptable for the pharmaceutical product to form a pharmaceutical product. The dosage forms include tablets, granules, capsules, pills, powders, liquids, suspensions, emulsions, syrups, elixirs, extracts and other oral preparations, or injections. Dosage forms using parenteral agents such as liquids, suppositories, ointments, patches, paps, lotions, etc. can be mentioned, but there are no particular restrictions, and appropriate selection should be made according to the purpose of treatment or prevention. Can be done.

Further, in the case of tablets, granules, pills, capsules and powders, additives such as excipients, binders, disintegrants and lubricants can be contained. Examples of the excipient include starch, carboxymethyl cellulose, sucrose, dextrin, cornstarch and the like.

Binders include crystalline cellulose, crystalline cellulose / carmellose sodium, methylcellulose, hydroxypropylcellulose, low-substituted hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, carmellose sodium, ethylcellulose, Carboxymethylethyl cellulose, hydroxyethylcellulose, wheat starch, rice starch, corn starch, potato starch, dextrin, pregelatinized starch, partially pregelatinized starch, hydroxypropyl starch, purulan, polyvinylpyrrolidone, aminoalkylmethacrylate copolymer E, aminoalkylmethacrylate copolymer RS , Methacrylic acid copolymer L, methacrylic acid copolymer, polyvinyl acetal diethylaminoacetate, polyvinyl alcohol, arabic rubber, arabic rubber powder, agar, gelatin, white serrac, tragant, purified sucrose, macrogol.

Disintegrants include crystalline cellulose, methyl cellulose, low-substituted hydroxypropyl cellulose, carmellose, carmellose calcium, carmellose sodium, croscarmellose sodium, wheat starch, rice starch, corn starch, potato starch, partially pregelatinized starch, hydroxy. Examples include propyl starch, sodium carboxymethyl starch and tragant.

Lubricants include wheat starch, rice starch, corn starch, stearic acid, calcium stearate, magnesium stearate, hydrous silicon dioxide, light anhydrous silicic acid, synthetic aluminum silicate, dry aluminum hydroxide gel, talc, alumine metasilicate. Examples thereof include magnesium acid, calcium hydrogen phosphate, anhydrous calcium hydrogen phosphate, sucrose fatty acid ester, waxes, hydrogenated vegetable oil, and polyethylene glycol.

In the case of liquids, syrups, suspensions, emulsions, and elixirs, in addition to commonly used inert diluents such as water and vegetable oils, colorants, flavoring agents, flavoring agents, etc. are added. It may be contained as an agent.

Further, in the case of an injection, an additive such as a suspension, an emulsion, or a solubilizer at the time of use can be contained. In the case of ointments and suppositories, additives such as fat, fatty oil, lanolin, vaseline, paraffin, wax, resin, plastic, base, glycols, higher alcohol, water, emulsifier, suspending agent and the like are added. Can be contained as. Further, in the case of a poultice, glycerin, water, a water-soluble polymer, a water-absorbing polymer and the like can be contained as additives. Further, in the case of a lotion agent, a solvent, an emulsifier, a suspending agent and the like can be contained as additives.

In addition, a product containing three components can be added to cosmetics. Examples of cosmetics include lotion, milky lotion, face wash, cleansing, beauty essence, cream, foundation, eyebrow, mascara, eyeshadow, eyeliner, lipstick, gloss, teak, white powder, manicure and the like. Further, as the form of cosmetics, forms such as liquid, cream, solid, stick, and powder can be adopted.

Further, the three-component compounded product may be blended in foods, beverages and the like. Examples of foods include breads, noodles, confectionery, processed meat products, processed seafood products, frozen foods, jellies, ice creams, dairy products, various seasonings and the like. In addition to general foods, it can also be added to foods for specified health use, quasi-drugs, health foods, and supplements. Examples of the beverage include soft drinks, milk beverages, alcoholic beverages, tea, tea beverages, coffee, fruit juice beverages, carbonated beverages, mineral waters, fruit / vegetable beverages and the like.
Further, the food or beverage containing the three-component compound may be in the same form as the orally-administered preparation such as tablets, capsules and syrup.

In addition, when producing foods and beverages containing a three-component compound, sweeteners, colorants, preservatives, thickeners, stabilizers, as necessary within the range that does not interfere with the effects of the present invention. Additives such as gelling agents, antioxidants, color formers, bleaching agents, emulsifiers, leavening agents, acidulants, brighteners, fragrances; solvents; oils may be added. One type of these additives may be used alone, or two or more types may be used in combination.

The ratio of the three-component compound to be blended in the food or beverage can be appropriately adjusted according to the purpose of use, but the proportion of the three-component compound to be blended in the food or beverage is preferably 0. It is .0001 to 80% by weight, more preferably 0.003 to 50% by weight, still more preferably 0.005 to 30% by weight.

Hereinafter, the present invention will be described with reference to examples, but the present invention is not limited thereto. Unless otherwise specified, parts and% in this embodiment are based on weight.

(Examination of nitric oxide production promoting effect)
Concentration (μg / mL) of human aortic smooth muscle cells in the medium for each of diosgenin, hirsutine, glycyrrhizin and a three-component formulation (diosgenin, hirsutine and glycyrrhizin in a 1: 1: 1 weight ratio). We evaluated changes in nitric oxide production in human aortic smooth muscle cells with an increase in hirsutine. The production of nitric oxide in the human aortic smooth muscle cells is an oxidation metabolite of nitric oxide nitrite _- index relative fluorescence intensity (RFU) of DAN fluorescent probe which emits fluorescence by reacting with (NO ²⁾ Evaluated to.

As a specific procedure, human Aortic Smooth Muscle Cells (HAoSMC); PromoCell) was cultured using a special medium (smooth muscle cell growth medium 2; PromoCell). ^{5 × 10 4} cells were seeded on a 24-well flat bottom plate and cultured to 80% confluent. Then, the medium was replaced with a medium containing 0.1, 1, 10 μg / mL of diosgenin, hirsutine, glycyrrhizin and a three-component compound, respectively, or a control medium containing no of these components (diosgenin, hirsutine and glycyrrhizin), and after 1 hour. The culture supernatant was collected. Total NO production was measured according to the standard protocol of the OxiSelect In VitroNitric Oxide (Nitrite / Nitrate) Assay Kit (CELL BIOLABS). The measurement results are shown in FIG.

We also examined the study by measuring the amount of nitric oxide synthase (NO synthase) gene.

Specifically, human Aortic Smooth Muscle Cells (HAoSMC); PromoCell) was cultured using a special medium (smooth muscle cell growth medium 2; PromoCell). ^{5 × 10 4} cells were seeded on a 24-well flat bottom plate and cultured to 80% confluent. Then, the medium was replaced with a medium containing 1,10,100 μg / mL of diosgenin, hirsutine, glycyrrhizin and a three-component compound, respectively, or a control medium containing no of these components (diosgenin, hirsutine and glycyrrhizin), and the cells were cultured after 1 hour. Qing was recovered. Total RNA was extracted with the RNeasy Mini Kit (QUIAGEN), reverse transcription reaction was performed with the OneStep RT-PCR Kit (NEB), and then the NO synthase gene and the constantly expressed GAPDH gene were amplified. rice field. The primers used for amplification are as follows.
NO synthase
5'-ACCATATTCCCCCAGAGGAC-3' (SEQ ID NO: 1)
5'-GAAGAGCTCAGGGTCATTGC-3'(SEQ ID NO: 2)
GAPDH
5'-AGGCAAGTCAAGGTCAAGCAAG-3' (SEQ ID NO: 3)
5'-ATGTTTGTGATGGGCGTGAACC-3' (SEQ ID NO: 4)

The difference from GAPDH (DeltaCt) of each sample was calculated by quantitative PCR, and the difference from the DeltaCt value (DeltaDeltaCt value) of the control sample without ingredients was obtained. The results are shown in FIG. 2 by multiplying the amount of mRNA of the control sample as the amount of mRNA expressed.

As shown in FIGS. 1 and 2, the production of nitric oxide in smooth muscle cells is increased by each component. In particular, its action is strong on diosgenin and glycyrrhizin, and the three-component compound is also increased in a concentration-dependent manner.
In addition, the mRNA level was increased in glycyrrhizin and the three-component compound.

(Examination of heat shock protein inducing ability)
Mouse nerve cells were used in the experiment. As mouse neurons, Neuro-2a (purchased from JCRB cell bank) was used, and a medium containing 10% FBS and 1% non-essential amino acids added to a commercially available medium (trade name D-MEM, purchased from Wako). Was cultured at 37 ° C. under _{the condition of 5% CO 2.} Immediately before the experiment, cells were collected with a 0.25% Trypsin / EDTA solution, and the number of cells was calculated by Countess (Invitrogen) after a trypan blue staining treatment. Then, the seeds were sown on a 24-well plate or a 96-well plate, precultured for 24 hours, and then used for the experiment.

Diosgenin, hirsutine, glycyrrhizin and a three-component compound (diosgenin, hirsutine and glycyrrhizin in a weight ratio of 1: 1: 1) were added to 0.1, 10, 100 μg / mL using a culture medium, respectively. A sample was obtained. Each sample was then added to mouse neurons, respectively. Then, the mRNA levels of Hsp32, Hsp47, Hsp70, gp96 and Hsp90 at 1 hour and 4 hours after the sample was added to the mouse nerve cells were measured by a quantitative PCR method.

The specific procedure of the quantitative PCR method was as follows. Total RNA was extracted from mouse neurons to which the sample was added using Trizol reagent (ambion). The extracted RNA was reverse transcribed into cDNA according to the method of PrimeScript RT Master Mix (Takara) and amplified by SYBR Premix EX Taq II (Takara). As the PCR reaction solution, 50μL (25μL SYBR Green Mix ( 2x), 1μL cDNA, 2μL primer pair mix (5 pmol/μL each primer), 22μL H 2 O) using the PCR reaction liquid, the reaction to 95 ° C. A cycle of 30 seconds was performed for one cycle, and a cycle of 5 seconds at 95 ° C. and a cycle of 30 seconds at 60 ° C. was performed under the conditions of 50 cycles. For RT-PCR, primers of various predetermined targets and housekeeping genes (GAPDH) described below were used.
Hsp32
5'-TCCGATGGGTCCTTACACTC-3'(SEQ ID NO: 5)
5'-TAAGGAAGCCAGCCAAGAGA-3' (SEQ ID NO: 6)
Hsp47
5'-CCCTGAAAGTCCCAGATCAA-3' (SEQ ID NO: 7)
5'-GGGAGAGGTTGGGATAGAGC-3'(SEQ ID NO: 8)
Hsp70
5'-AGTGGTGGCCACTAATGGAG-3' (SEQ ID NO: 9)
5'-CAATCCTTGCTTGATGCTGA-3' (SEQ ID NO: 10)
Hsp90
5'-TGGACAGACAAACATGGAGAG-3' (SEQ ID NO: 11)
5'-AGACAGGAGCCAGTTTCAT -3'(SEQ ID NO: 12)
gp96
5'-TGGGAAGAGGTTCCAGAATG-3'(SEQ ID NO: 13)
5'-GTTGCCAGACCATCCGTACT -3'(SEQ ID NO: 14)
GAPDH
5'-AGGCAAGTCAAGGTCAAGCAAG-3' (SEQ ID NO: 3)
5'-ATGTTTGTGATGGGCGTGAACC-3' (SEQ ID NO: 4)

In addition, the results 1 hour after each sample was added to the mouse nerve cells are shown in FIG. 3, and the results 4 hours after each are shown in FIG. The heat shock proteins increased in each component are shown in Table 1. In addition, Table 2 shows the role of the heat shock protein whose expression was increased.

From the above, it was shown that the combination of the three components (diosgenin, hirsutine and glycyrrhizin) has the ability to produce nitric oxide and the ability to induce several heat shock proteins. From this, it can be said that the three-ingredient combination product has an effective effect on menopausal disorders and anti-aging, regardless of gender.

Claims (15)

A heat shock protein inducer containing at least one component of diosgenin, hirsutine and glycyrrhizin. The heat shock protein inducer according to claim 1, which contains three components of diosgenin, hirsutine and glycyrrhizin. The heat shock protein inducer according to claim 2, wherein the amount of diosgenin is 50% by weight or less, the amount of hirsutine is 50% by weight or less, and the amount of glycyrrhizin is 50% by weight or less. The heat shock protein inducer according to any one of claims 1 to 3, wherein the heat shock protein is at least one of gp96, Hsp32, Hsp47, Hsp70, and Hsp90. An anti-menopausal agent containing the heat shock protein inducer according to any one of claims 1 to 4. An anti-aging agent containing the heat shock protein inducer according to any one of claims 1 to 4. A cosmetic product containing the heat shock protein inducer according to any one of claims 1 to 4. A food or beverage containing the heat shock protein inducer according to any one of claims 1 to 4. A nitric oxide production promoter containing at least one component of diosgenin or glycyrrhizin. The nitric oxide production promoter according to claim 9, which comprises the three components of diosgenin, hirsutine and glycyrrhizin. The nitric oxide production promoter according to claim 10, wherein the amount of diosgenin is 50% by weight or less, the amount of hirsutine is 50% by weight or less, and the amount of glycyrrhizin is 50% by weight or less. An anti-menopausal agent containing the nitric oxide production promoter according to any one of claims 9 to 11. An anti-aging agent containing the nitric oxide production promoter according to any one of claims 9 to 11. A cosmetic product containing the nitric oxide production promoter according to any one of claims 9 to 11. A food or beverage containing the nitric oxide production promoter according to any one of claims 9 to 11.

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