JP2022111283A - Novel collagen reuse promoting effect agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a fragmented collagen metabolism promoter containing as an active ingredient a component having excellent fragmented collagen metabolism promoting action which is found by us, and to provide a wrinkle improving composition and a composition of the prevention/improvement of arthritis which contain the fragmented collagen metabolism promoter.

SOLUTION: An Endo180 activation component is used as an active ingredient of the fragmented collagen metabolism promoter of the present invention. Preferably, the Endo180 activation component is a plant-derived component, and the plant-derived component is one or more selected from the group consisting of stevia extract, rubusoside, stevioside, rebaudioside A, and rebaudioside C, preferably. Provided are a composition for a wrinkle improvement and the composition for arthritis prevention/improvement containing the aforementioned fragmented collagen metabolism enhancer, and also food and drinks for wrinkle improvement and food and drinks for the preventive/improving of arthritis which contains a wrinkle improving composition and arthritis preventing/improving composition respectively.

SELECTED DRAWING: None

COPYRIGHT: (C)2022,JPO&INPIT

Description

TECHNICAL FIELD The present invention relates to a fragmented collagen metabolism-promoting agent, and more particularly to a fragmented collagen metabolism-promoting agent containing an Endo180 activating ingredient as an active ingredient. Furthermore, the present invention also relates to a composition for improving wrinkles and a composition for preventing and improving arthritis containing a fragmented collagen metabolism-promoting agent.

Quantitative and qualitative changes in dermal collagen fiber structure have been confirmed in photoaging caused by long-term exposure to ultraviolet rays, and are thought to be involved in wrinkle formation. Homeostasis of collagen fiber structure is maintained by metabolic balance due to repeated synthesis and degradation of collagen fibers. In the degradation process, collagen is fragmented by matrix metalloproteases, and the fragmented collagen is incorporated into fibroblasts and degraded. The involvement of the protein collagen receptor "Endo180" is known (Non-Patent Document 1).

Endo180, also known as mannose receptor C2 (MRC2), urokinase plasminogen activator receptor-associated protein (uPARAP) and CD280, is a recycled endocytic receptor that binds ligands such as collagen. Then, it is taken up into cells and induced to be degraded in endosomes.

Endo180 has homology with the macrophage mannose receptor family, mannose receptor, phospholipase A2, DEC-205/MR6, and others. Endo180 is unique among the family of mannose receptors in that it is targeted from the plasma membrane to recycling endosomes, rather than to the late endosomal/lysosomal compartment, and it is a mannose receptor for cell migration and collagen for intracellular degradation. It functions in cell motility and extracellular matrix remodeling by promoting the uptake of As uPARAP, it forms a triple complex with urokinase-type plasmin activator (uPA) and urokinase-type plasmin activator receptor (uPAR), and participates in the generation of plasmin from plasminogen. Endo180 is localized in clathrin-covered pits on the cell surface, endosomes, etc., and is mainly expressed in fibroblasts, endothelial cells, and macrophages. As tissues, it is mainly expressed in lungs, blood vessels, bones and skin (see Patent Document 1).

On the other hand, although the dermis of the skin is also a tissue rich in collagen, there are few reports on the involvement of Endo180 in the remodeling of collagen fibers in the dermis. So far, it has been known that in photoaged skin, Endo180 expression is decreased in fibroblasts, which accompanies the accumulation of fragmented collagen in the dermal matrix (Non-Patent Document 2). It is also known that fragmented collagen induces oxidative stress in fibroblasts (Non-Patent Document 3). Furthermore, it is also known that IL-1α produced by keratinocytes is involved in the mechanism of UV-induced decrease in Endo180 expression (Non-Patent Document 4). Based on these findings, Endo180 is believed to be involved in the formation of photoaged skin (eg, formation of wrinkles) caused by long-term exposure to ultraviolet rays. As a mechanism, when the expression of Endo180 is decreased in fibroblasts, the uptake of fragmented collagen is suppressed, which inhibits regeneration of normal collagen fiber structure, and fragmented collagen accumulates around the cells. It is thought to induce dysfunction of fibroblasts.

Therefore, for example, if Endo180 expression can be promoted in fibroblasts, it will promote the reuse of fragmented collagen through metabolism and the regeneration of normal collagen fiber structure, thereby suppressing the formation of photoaged skin and the resulting wrinkles. is considered possible.

Japanese Patent Publication No. 2012-521217

J. Cell Sci., 2009, vol.122, pp.4042-4048 J. Dermatol. Sci., 2013, vol.70, pp.42-48 Am. J. Pathol., 2009, vo.174, pp.101-114 Abstracts of the 40th Annual Meeting of the Japanese Cosmetic Science Society, 2015, p.16

The present invention finds a component having an excellent fragmented collagen metabolism promoting action, a fragmented collagen metabolism promoting agent containing the fragmented collagen metabolism promoting agent as an active ingredient, and a wrinkle-improving composition and arthritis prevention containing the fragmented collagen metabolism promoting agent. - Aimed at providing an improvement composition.

In order to solve the above problems, the present inventors provide the following inventions.
[1] A fragmented collagen metabolism-promoting agent characterized by containing an Endo180 activating ingredient as an active ingredient.
[2] The fragmented collagen metabolism promoting agent according to [1], wherein the Endo180 activating component is a plant-derived component.
[3] The plant-derived component is one or more selected from the group consisting of stevia extract, rubusoside, stevioside, rebaudioside A and rebaudioside C [1] The fragmented collagen metabolism promoting agent according to [2].
[4] A wrinkle-improving composition comprising the fragmented collagen metabolism-promoting agent of [1] to [3].
[5] The wrinkle improvement according to [4], characterized by containing one or more components selected from the group consisting of collagen production promoters, blood flow promoters, collagen, proteins, peptides and amino acids. composition.
[6] [4] A wrinkle-improving food or drink comprising the wrinkle-improving composition of [5].
[7] A composition for preventing/ameliorating arthritis, comprising the fragmented collagen metabolism-promoting agent of [1] to [3].
[8] Contains one or more components selected from collagen production promoters, anti-inflammatory agents, blood flow promoters, collagen, proteins, peptides, amino acids, black ginger, glucosamine, N-acetylglucosamine, and chondroitin The composition for preventing/ameliorating arthritis according to [7], characterized by:
[9] A food or drink for preventing/ameliorating arthritis, comprising the composition for preventing/ameliorating arthritis according to [7][8].

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a fragmented collagen metabolism promoting agent having excellent effects. Furthermore, by containing the fragmented collagen metabolism promoting agent, it is possible to provide a composition for improving wrinkles and a composition for preventing/improving arthritis, which are excellent in promoting the metabolism of fragmented collagen.

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described below.
The fragmented collagen metabolism promoting agent of the present embodiment contains an Endo180 activating ingredient as an active ingredient. In addition, the composition for improving wrinkles and the composition for preventing/improving arthritis of the present embodiment contain the fragmented collagen metabolism promoting agent.

In this embodiment, the Endo180 activating ingredient is preferably a plant-derived ingredient. Such a plant-derived component is not particularly limited as long as it has an Endo180 activating action, but one selected from the group consisting of stevia extract, rubusoside, stevioside, rebaudioside A and rebaudioside C, or Two or more are preferred.

[Stevia extract]
In the present embodiment, the "stevia extract" includes an extract obtained from stevia as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or a crude purification thereof. either the product or the purified product.

Stevia (scientific name: Stevia rebaudiana) is a perennial herb belonging to the genus Stevia of the Asteraceae family distributed in the tropical to temperate regions of the Americas, and can be easily obtained from these regions. Stevia extracts are conventionally used as sweeteners and the like. Constituent parts of stevia that can be used as raw materials for extraction include, for example, leaves, branches, stems, flowers, buds, seeds, aerial parts, whole plants, and mixtures thereof, but are preferably used. is the leaf part.

The extract from the above plant can be obtained by drying the raw material for extraction, either as it is or pulverizing it using a crusher, and subjecting it to extraction with an extraction solvent. Drying may be performed in the sun or using a commonly used dryer. In addition, it may be used as an extraction raw material after being subjected to pretreatment such as degreasing with a non-polar solvent such as hexane. By performing pretreatment such as degreasing, the extraction treatment with a polar solvent can be efficiently performed.

As the extraction solvent, it is preferable to use a polar solvent, for example, water, a hydrophilic organic solvent, etc., which are used alone or in combination of two or more at room temperature or at a temperature below the boiling point of the solvent. is preferred.

Water that can be used as an extraction solvent includes pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, as well as those subjected to various treatments. Treatments applied to water include, for example, purification, heating, sterilization, filtration, ion exchange, osmotic adjustment, buffering, and the like. Therefore, water that can be used as an extraction solvent in the present embodiment includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered physiological saline, and the like.

Hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene. Examples include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

When using a mixture of two or more polar solvents as the extraction solvent, the mixture ratio can be adjusted as appropriate. For example, when a mixture of water and a lower aliphatic alcohol is used as the extraction solvent, the mixing ratio of water and the lower aliphatic alcohol is preferably 9: 1 to 1: 9 (volume ratio), More preferably, it is 7:3 to 2:8 (volume ratio). When using a mixture of water and lower aliphatic ketone, the mixing ratio of water and lower aliphatic ketone is preferably 9:1 to 2:8 (volume ratio). When using a mixed solution with a hydric alcohol, the mixing ratio of water and polyhydric alcohol is preferably 5:5 to 1:9 (volume ratio).

The extraction treatment is not particularly limited as long as the soluble components contained in the raw material for extraction can be eluted into the extraction solvent, and can be carried out according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent of 5 to 15 times the amount (mass ratio) of the extraction raw material, and the soluble components are extracted at room temperature or under reflux heating, and then filtered to remove the extraction residue. You can get the liquid. A paste-like concentrate is obtained by distilling off the solvent from the obtained extract, and a dried product is obtained by further drying the concentrate.

The stevia extract obtained as described above can be used as it is as an Endo180 activating component, but it is easier to use as a concentrated solution or dried product.

[Rubusoside, stevioside, rebaudioside A, rebaudioside C]
Rubusoside (13β-(β-D-glucopyranosyloxy) Kaur-16-en-18-acid β-D-glucopyranosyl), Stevioside (13-[(2-O-β-D-glucopyranosyl-β-D -glucopyranosyl)oxy]kaur-16-ene-18-acid β-D-glucopyranosyl), rebaudioside A (13-[(2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl -β-D-glucopyranosyl)oxy]kaur-16-ene-18-acid β-D-glucopyranosyl), and rebaudioside C (13-[[2-O-(6-deoxy-α-L-mannopyranosyl) )-3-O-(β-D-glucopyranosyl)-β-D-glucopyranosyl]oxy]kaur-16-en-18-acid β-D-glucopyranosyl) are both steviol obtainable from plant extracts It is a glycoside.

A plant extract containing rubusoside and a plant extract containing at least one of stevioside, rebaudioside A and rebaudioside C can be obtained by methods commonly used for plant extraction. Plants containing rubusoside include sweet tea (scientific name: Rubus suavissimus), and plants containing stevioside, rebaudioside A or rebaudioside C include the aforementioned stevia (scientific name: Stevia rebaudiana). but not limited to these.

Sweet tea (Rubus suavissimus) is a plant belonging to the genus Rubus of the family Rosaceae, grows naturally in southern China, and can be easily obtained from these regions. Tien tea extract is conventionally used as an antiallergic agent and the like. The part that can be used as an extraction raw material is not particularly limited, but the leaf part is suitable.

Tien tea extract from which rubusoside can be obtained can be obtained by using Tien tea as an extraction raw material and extracting it in the same manner as the stevia extract described above. As the stevia extract from which stevioside, rebaudioside A or rebaudioside C can be obtained, the stevia extract described above can be preferably used.

The method for isolating and purifying rubusoside, stevioside, rebaudioside A and rebaudioside C from the extract obtained as described above, the concentrate of the extract, or the dried product of the extract is particularly It is not limited and can be carried out by a conventional method. For example, the extract is dissolved in a developing solvent and subjected to column chromatography using a porous material such as silica gel or alumina, or a porous resin such as styrene-divinylbenzene copolymer or polymethacrylate to obtain rubusoside and stevioside. , a method of collecting a fraction containing rebaudioside A or rebaudioside C, and the like. In this case, the developing solvent may be appropriately selected according to the stationary phase used. For example, when the extract is separated by normal phase chromatography using silica gel as the stationary phase, the developing solvent is chloroform:methanol=95: 5 and the like. Furthermore, fractions containing rubusoside, stevioside, rebaudioside A or rebaudioside C obtained by column chromatography were subjected to reversed-phase silica gel chromatography using ODS, recrystallization, and liquid-liquid countercurrent extraction. Any organic compound purification means such as column chromatography using an ion exchange resin may be used for purification.

In addition, rubusoside, stevioside, rebaudioside A, or rebaudioside C used in this embodiment may be commercially available products.

[Fragmented Collagen Metabolism Promoter]
Since the stevia extract, rubusoside, stevioside, rebaudioside A and rebaudioside C obtained as described above have an excellent Endo180 activating action, they are effective ingredients of the fragmented collagen metabolism promoting agent. It is suitable as The fragmented collagen metabolism promoting agent of the present embodiment can be used in a wide range of applications such as pharmaceuticals, quasi-drugs, cosmetics, and food and drink.

Here, the Endo180 activating action of stevia extract, rubusoside, stevioside, rebaudioside A and rebaudioside C is exhibited based on the Endo180 mRNA expression promoting action and/or Endo180 production promoting action. However, the Endo180 activating action of stevia extract, rubusoside, stevioside, rebaudioside A and rebaudioside C is not limited to the Endo180 activating action exhibited based on the above action.

In addition, stevia extract, rubusoside, stevioside, rebaudioside A and rebaudioside C are Endo180 activators and Endo180 activators using their Endo180 activating action, Endo180 mRNA expression promoting action and Endo180 production promoting action, respectively. You may use it as an active ingredient of an Endo180 mRNA expression promoter or an Endo180 production promoter.

The fragmented collagen metabolism promoting agent of the present embodiment may consist of only the Endo180 activating component, or may be a formulation of the Endo180 activating component.

The fragmented collagen metabolism-promoting agent of the present embodiment can be prepared in the form of powder, granules, tablets, liquid, etc., according to a conventional method, using a pharmaceutically acceptable carrier such as dextrin, cyclodextrin, or any other adjuvant. It can be formulated into any dosage form. At this time, as auxiliary agents, for example, excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents and the like can be used. Fragmented collagen metabolism-promoting agents can be used by blending with other compositions (for example, food and drink for improving wrinkles, food and drink for preventing and improving arthritis, etc., which will be described later). It can be used as an agent or the like.

When the fragmented collagen metabolism promoting agent of the present embodiment is formulated, the content of the Endo180 activating component is not particularly limited, and can be appropriately set according to the purpose.

In the fragmented collagen metabolism promoting agent of the present embodiment, if necessary, other natural product-derived components having a fragmented collagen metabolism promoting effect may be blended together with the Endo180 activating component and used as active ingredients. can be done.

Methods for administering the fragmented collagen metabolism-promoting agent of the present embodiment to patients include transdermal administration, oral administration, and the like. Depending on the type of disease, a suitable method for prevention, treatment, etc., may be appropriately selected. Just do it. In addition, the dose of the fragmented collagen metabolism-promoting agent of the present embodiment may also be appropriately increased or decreased depending on the type and severity of disease, individual patient differences, administration method, administration period, and the like.

The fragmented collagen metabolism-promoting agent of the present embodiment has the Endo180 activating action of the active ingredient, the Endo180 activating component, particularly stevia extract, rubusoside, stevioside, rebaudioside A, and rebaudioside C. Activation of Endo180 in fibroblasts through mRNA expression promoting action or Endo180 production promoting action, thereby promoting uptake of fragmented collagen by endocytosis, promoting regeneration of normal collagen fiber structure, formation of wrinkles, etc. It is possible to prevent, treat or ameliorate skin photoaging symptoms such as In addition, the fragmented collagen metabolism-promoting agent of the present embodiment promotes the regeneration of normal collagen fiber structure, thereby treating rheumatoid arthritis, osteoarthritis, septic arthritis, gouty arthritis, traumatic arthritis, osteoarthritis, and the like. It can be used for purposes such as prevention, treatment or improvement of arthritis.

In addition to the uses described above, the fragmented collagen metabolism-promoting agent of the present embodiment can be used for all uses that are significant in exhibiting an Endo180 activating action, an Endo180 mRNA expression promoting action, or an Endo180 production promoting action. can be done. For example, the fragmented collagen metabolism promoting agent of the present embodiment, or the aforementioned Endo180 activating agent, Endo180 mRNA expression promoting agent, or Endo180 production promoting agent has an Endo180/uPARAP activating action, Endo180 mRNA expression promoting action, or Endo180 production Through stimulatory action, it can promote the production of plasmin from plasminogen.

Since the fragmented collagen metabolism-promoting agent of the present embodiment has an excellent fragmented collagen metabolism-promoting effect, it can be used in other compositions such as skin external preparations (for example, wrinkle-improving compositions, wrinkle-improving food and drink products, which will be described later). composition for preventing/improving arthritis, food and drink for preventing/improving arthritis). In this case, the Endo180 activating component, especially the stevia extract, rubusoside, stevioside, rebaudioside A or rebaudioside C may be blended as it is, or fragmented formulated from the Endo180 activating component. A collagen metabolism accelerator may be blended.

In addition, since the fragmented collagen metabolism-promoting agent of the present embodiment has an excellent fragmented collagen metabolism-promoting action, it can be suitably used as a reagent for research on these mechanisms of action.

The fragmented collagen metabolism-promoting agent of the present embodiment is preferably applied to humans. , dogs, cats, cows, pigs, monkeys, etc.).

[Composition for improving wrinkles, composition for preventing and improving arthritis]
Since the Endo180 activating component has an excellent fragmented collagen metabolism promoting effect, it is suitable for blending in a composition for improving wrinkles or a composition for preventing/improving arthritis. In this case, the Endo180 activating component, especially the stevia extract, rubusoside, stevioside, rebaudioside A or rebaudioside C may be blended as it is, or fragmented collagen formulated from the Endo180 activating component. A metabolism accelerator may be added.

By blending the above-mentioned Endo180 activating component or fragmented collagen metabolism promoting agent, the composition can be endowed with Endo180 activating action, Endo180 mRNA expression promoting action or Endo180 production promoting action, and is used for wrinkle improvement and arthritis prevention. • Can be particularly suitable for remediation applications.

The dosage form of the composition for improving wrinkles and the composition for preventing/improving arthritis according to the present embodiment is not particularly limited and can be appropriately selected depending on the intended purpose. mentioned.
In addition, the dosage form of the composition for improving wrinkles and the composition for preventing/improving arthritis according to the present embodiment is not particularly limited, and can be appropriately selected according to the purpose. Examples include tablets, powders, and capsules. , granules, extracts, syrups and the like; parenteral administration agents such as injections, drops, and suppositories; and topical agents such as ointments.

The wrinkle-improving composition according to the present embodiment can be particularly suitably used as an external preparation for skin. There are no restrictions on the category of topical agents for skin, and they include a wide range of transdermal skin cosmetics, quasi-drugs, pharmaceuticals, and the like.

When the wrinkle-improving composition and the arthritis-preventing/improving composition of the present embodiment are in the form of an orally administered agent, a parenterally administered agent, an externally administered agent, etc., the Endo180 activating ingredient, especially stevia extract, rubusoside , Stevioside, Rebaudioside A, and Rebaudioside C, as long as they do not interfere with the fragmented collagen metabolism-promoting action, main agents and auxiliary agents used in the production of usual oral administration agents, parenteral administration agents, external agents, etc. Or other ingredients such as astringents, bactericidal/antibacterial agents, whitening agents, UV absorbers, moisturizing agents, cell activators, anti-inflammatory agents, antioxidants/active oxygen removers, oils and fats, waxes, hydrocarbons , fatty acids, alcohols, esters, surfactants, fragrances and the like may be used in combination. Such combination may result in a more generalized product and may provide greater benefits than would normally be expected due to synergy between the other active ingredients used in combination.

In addition, the wrinkle-improving composition according to the present embodiment includes one or two selected from the group consisting of a collagen production promoter, a blood flow promoter, collagen, a protein, a peptide, and an amino acid, together with the fragmented collagen metabolism promoter. It is preferred to contain more than one species. These components can bring about a wrinkle-improving effect from a viewpoint different from the fragmented collagen metabolism-promoting effect. For example, by promoting the production of new collagen, the collagen production promoting agent cooperates with the fragmented collagen metabolism promoting action of the fragmented collagen metabolism promoting agent to promote the regeneration of normal collagen fiber structure and wrinkles. It is possible to more effectively prevent, treat or improve photoaging symptoms of the skin such as the formation of

In addition, the composition for preventing/improving arthritis according to the present embodiment contains, together with the fragmented collagen metabolism promoting agent, a collagen production promoting agent, an anti-inflammatory agent, a blood flow promoting agent, collagen, protein, peptide, amino acid, black ginger, It preferably contains one or more components selected from glucosamine, N-acetylglucosamine, and chondroitin. These components can bring about arthritis prevention/improvement effects from a different viewpoint from fragmented collagen metabolism promoting effects.

[Food and drink for improving wrinkles, food and drink for preventing and improving arthritis]
The wrinkle-improving composition provides the excellent fragmented collagen metabolism promoting action of the Endo180 activating component, especially stevia extract, rubusoside, stevioside, rebaudioside A, and rebaudioside C. It is suitable for blending in food and drink for improvement. Similarly, the composition for preventing/ameliorating arthritis is suitable for blending in foods and drinks for preventing/ameliorating arthritis. In this case, the above-mentioned Endo180 activating ingredients, especially stevia extract, rubusoside, stevioside, rebaudioside A or rebaudioside C, may be blended as they are, or these ingredients may be formulated to promote fragmented collagen metabolism. agents may be added.

Here, “food and drink” refers to those that are less likely to harm human health and are taken orally or through the gastrointestinal tract in normal social life. It is not limited to categories such as Therefore, the "wrinkle-improving food and drink" and the "arthritis-preventing and improving food and drink" in the present embodiment include orally ingested general foods, health foods (functional foods and drinks), health functional foods (for specified health food, food with nutrient function claims), quasi-drugs, pharmaceuticals, etc. The food and drink for improving wrinkles or the food and drink for preventing/improving arthritis according to the present embodiment contains the above-mentioned Endo180 activating ingredients, especially stevia extract, rubusoside, stevioside, rebaudioside A, and It is preferably a food or drink that can display the favorable effects of Baudioside C, and is a food with health claims (food for specified health uses, food with function claims, food with nutrient claims), quasi-drug, and drug. is particularly preferred.

When the wrinkle-improving composition or arthritis-preventing/improving composition is incorporated into food or drink, the amount of the active ingredient to be incorporated therein can be appropriately changed in consideration of the purpose of use, symptoms, gender, etc. Considering the general intake of the food and drink to which the extract is added, it is preferable that the intake of the extract per adult is about 1 to 1000 mg per day. When the food or drink to be added is a granular, tablet or capsule food or drink, the amount of the composition for improving wrinkles or the composition for preventing or improving arthritis is usually 0.1 with respect to the food or drink to be added. It is up to 100% by mass, preferably 5 to 100% by mass.

The food and drink for improving wrinkles or the food and drink for preventing and improving arthritis of the present embodiment may be blended with any food or drink that does not interfere with the activity of the Endo180 activating ingredient. It may be a dietary supplement containing the ingredients as a main ingredient.

When producing the food and drink for improving wrinkles or the food and drink for preventing and improving arthritis of the present embodiment, for example, sugars such as dextrin and starch; proteins such as gelatin, soybean protein and corn protein; alanine, glutamine, isoleucine and the like. polysaccharides such as cellulose and gum arabic; oils and fats such as soybean oil and medium-chain triglycerides.

The food and drink in which the wrinkle-improving composition or the arthritis-preventing/improving composition can be blended is not particularly limited, but specific examples thereof include soft drinks, carbonated drinks, nutritional drinks, fruit drinks, lactic acid drinks, Concentrated undiluted solutions and powders for preparation of these beverages); Frozen desserts such as ice cream, ice sherbet, and shaved ice; Confectionery such as chewing gum, candy, gum, chocolate, tablets, snacks, biscuits, jellies, jams, creams, and baked goods; Processed marine and livestock foods such as fish cakes, hams, and sausages; Dairy products such as processed milk and fermented milk Oils and fats such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, and dressings; Seasonings such as sauces and sauces; Soups, stews, salads, side dishes, pickles; Health and nutrition in various forms Supplementary foods; tablets, capsules, drinks and the like. When the wrinkle-improving composition or the arthritis-preventing/improving composition is blended into these foods and drinks, commonly used auxiliary raw materials and additives can be used in combination.

The present invention will be specifically described below with reference to test examples, but the present invention is not limited to the following examples. In the following test examples, the test samples were stevia extract (product name: "Jun Marumilon", manufactured by Maruzen Pharmaceutical Co., Ltd., sample 1), rubusoside (manufactured by Maruzen Pharmaceutical Co., Ltd., sample 2), stevioside (manufactured by Maruzen Pharmaceutical Co., Ltd., Sample 3), rebaudioside C (manufactured by Maruzen Pharmaceutical Co., Ltd., sample 4), and rebaudioside A (manufactured by Maruzen Pharmaceutical Co., Ltd., sample 5) were used.

[Test Example 1] Endo180 mRNA expression promoting effect test Endo180 mRNA expression promoting effect was tested for the natural product-derived components (Samples 1 to 5) as follows.

Human normal skin fibroblasts (NB1RGB) were cultured using 10% FBS-containing Dulbecco's MEM, and then the cells were collected by trypsinization. The recovered cells were diluted with FBS-free Dulbecco's MEM to a cell density of 1.0×10 ⁵ cells/mL, then seeded in 6-well microplates at 2 mL per well and cultured overnight.

After the culture was completed, the medium was removed, and 2 mL of test samples (Samples 1 to 5, see Table 1 below for sample concentrations) dissolved in FBS-free Dulbecco's MEM was added to each well and cultured for 24 hours. As a control, FBS-free Dulbecco's MEM to which no sample was added was used and cultured in the same manner. After culturing, the medium was removed, total RNA was extracted with ISOGEN II (manufactured by Nippon Gene, Cat. No. 311-07361), and the amount of each RNA was measured with a spectrophotometer so as to reach 100 ng/μL. Total RNA was prepared at .

Using this total RNA as a template, the expression level of mRNA was measured for Endo180 and GAPDH as an internal standard. Detection was performed by real-time 2-step RT-PCR reaction with TaKaRa SYBR PrimeScript ^™ RT-PCR Kit (Perfect Real Time) (code No. RR063A) using a real-time PCR device Smart Cycler (manufactured by Cepheid). The expression level of Endo180 mRNA was corrected with the value of GAPDH based on total RNA preparations prepared from cells cultured with "test sample added" and "test sample not added". From the obtained values, the Endo180 mRNA expression promotion rate (%) was calculated according to the following formula.

Endo180 mRNA expression promotion rate (%) = A/B x 100
Each term in the formula represents the following.
A: Correction value with addition of test sample B: Correction value with no addition of test sample Table 1 shows the results.

As shown in Table 1, stevia extract (sample 1), rubusoside (sample 2), stevioside (sample 3), rebaudioside C (sample 4), and rebaudioside A (sample 5) was also confirmed to have an excellent Endo180 mRNA expression promoting effect.

[Test Example 2] Endo180 production promoting action test The Endo180 production promoting action of the natural product-derived components (Samples 1 to 5) was tested as follows.

Human normal skin fibroblasts (NB1RGB) were cultured using 10% FBS-containing Dulbecco's MEM, and then the cells were collected by trypsinization. After diluting the recovered cells with FBS-free Dulbecco's MEM to a cell density of 1.0×10 ⁵ cells/mL, 100 μL of each well was seeded in a 96-well microplate and cultured overnight.

After the culture was completed, the medium was removed, and 100 μL of test samples (Samples 1 to 5, see Table 2 below for sample concentrations) dissolved in FBS-free Dulbecco's MEM were added to each well and cultured for 24 hours. As a control, FBS-free Dulbecco's MEM to which no sample was added was used and cultured in the same manner. After culturing, the amount of Endo180 present on the cell surface of each well was measured by ELISA. In addition, a standard curve was prepared using the Endo180 recombinant protein, and the amount of Endo180 produced was calculated. From the measurement results, the Endo180 production promotion rate (%) was calculated by the following formula.

Endo180 production promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Endo180 production amount with addition of test sample B: Endo180 production amount with no sample addition

Further, nuclear staining was performed using Hoechst 33342, and the amount of fluorescence was measured for cell counting. The amount of Endo180 produced per cell was calculated as a corrected value by dividing the amount of Endo180 produced using the calibration curve by the fluorescence wavelength (Ex: 350 nm/Em: 460 nm) of Hoechst 33342. Based on the obtained corrected values, the rate of promotion of Endo180 production (%) per cell was calculated according to the following formula.

Endo180 production promotion rate per cell (%) = C / D × 100
Each term in the formula represents the following.
C: Endo180 production amount per cell with test sample added D: Endo180 production amount per cell without test sample added Tables 2 and 3 show the results.

Stevia extract (Sample 1), Rubusoside (Sample 2), Stevioside (Sample 3), Rebaudioside C (Sample 4), and Rebaudioside A (Sample 5), as shown in Tables 2 and 3. was confirmed to have an excellent Endo180 production promoting effect.

[Formulation Example 1]
A milky lotion was prepared by a conventional method according to the following composition.
Stevia extract 0.01g
Jojoba oil 4.00g
1,3-butylene glycol 3.00 g
Arbutin 3.00g
Polyoxyethylene cetyl ether (20E.O.) 2.50g
2.00 g olive oil
2.00 g of squalane
Cetanol 2.00g
Glyceryl monostearate 2.00g
Polyoxyethylene sorbitan oleate (20E.O.) 2.00g
Methyl paraoxybenzoate 0.15g
Stearyl glycyrrhizinate 0.10g
Huangji extract 0.10g
Dipotassium glycyrrhizinate 0.10g
Ginkgo biloba extract 0.10g
Conchiolin 0.10g
Phellodendron bark extract 0.10g
Chamomile extract 0.10g
Perfume 0.05g
Remainder of purified water (total amount is 100 g)

[Formulation Example 2]
A cream having the following composition was produced by a conventional method.
Rubusoside 0.05g
Kujin extract 0.1g
Scutellaria root extract 0.1g
Liquid paraffin 5.0g
Bleached beeswax 4.0g
Squalane 10.0g
Cetanol 3.0g
Lanolin 2.0g
1.0 g of stearic acid
Polyoxyethylene sorbitan oleate (20E.O.) 1.5g
Glyceryl monostearate 3.0g
Oil-soluble licorice extract 0.1g
1,3-butylene glycol 6.0 g
Methyl paraoxybenzoate 1.5g
Perfume 0.1g
Remainder of purified water (total amount is 100 g)

[Formulation Example 3]
A beauty essence having the following composition was prepared by a conventional method.
Stevioside 0.01g
Chamomile extract 0.1g
carrot extract 0.1g
xanthan gum 0.3g
0.1 g of hydroxyethyl cellulose
Carboxyvinyl polymer 0.1 g
1,3-butylene glycol 4.0 g
Dipotassium glycyrrhizinate 0.1g
2.0 g of glycerin
Potassium hydroxide 0.25g
Perfume 0.01g
Preservative (methyl paraoxybenzoate) 0.15g
2.0 g of ethanol
Remainder of purified water (total amount is 100 g)

[Formulation Example 4]
A hair tonic having the following composition was produced by a conventional method.
Rebaudioside A 0.4g
Tocopherol acetate Appropriate amount Cephalatine 0.002g
Isopropylmethylphenol 0.1g
Sodium hyaluronate 0.15g
15.0 g of glycerin
Ethanol 15.0g
Perfume Appropriate amount Chelating agent (edetate sodium) Appropriate amount Preservative (hinokitiol) Appropriate amount Solubilizer (polyoxyethylene cetyl ether) Appropriate amount Purified water Balance

[Formulation Example 5]
A shampoo having the following composition was produced by a conventional method.
Rebaudioside C 0.5g
Marjoram extract 1.0g
Plum fruit part extract 0.2g
Coconut fatty acid methyl taurate sodium 10.0 g
Coconut fatty acid amidopropyl betaine 10.0g
Sodium polyoxyethylene alkyl ether sulfate 20.0 g
Coconut fatty acid diethanolamide 4.0 g
Propylene glycol 2.0 g
Fragrance Appropriate amount Purified water Remainder (Total amount is 100g)

[Formulation Example 6]
A tablet having the following composition was produced by a conventional method.
Stevia extract 5.0 mg
Dolomite (containing 20% calcium and 10% magnesium) 83.4mg
Casein phosphopeptide 16.7mg
Vitamin C 33.4mg
Maltitol 136.8mg
Collagen 12.7mg
Sucrose fatty acid ester 12.0mg

[Formulation Example 7]
An oral liquid preparation having the following composition was prepared by a conventional method.
<Composition in 1 ampoule (100 mL per bottle)>
Rebaudioside A 0.3% by mass
Hihatsu extract 0.15% by mass
Sorbit 12.0% by mass
Sodium benzoate 0.1% by mass
Perfume 1.0% by mass
Calcium sulfate 0.5% by mass
Remainder of purified water (100% by mass)

The fragmented collagen metabolism-promoting agent of the present invention can greatly contribute to the prevention, treatment, or amelioration of skin photoaging symptoms such as wrinkle formation; the prevention, treatment, or amelioration of various types of arthritis;

Claims (7)

An Endo180 activator comprising one or more selected from the group consisting of stevia extract, rubusoside, stevioside, rebaudioside A and rebaudioside C as an active ingredient. A wrinkle-improving composition based on Endo180 activation use, characterized in that it contains the Endo180 activator according to claim 1 . 3. The anti-wrinkle composition according to claim 2, comprising one or more components selected from the group consisting of collagen production promoters, blood flow promoters, collagen, proteins, peptides and amino acids. . 4. A wrinkle-improving food or drink based on Endo180 activating use, comprising the wrinkle-improving composition according to claim 2 or 3. A composition for preventing/ameliorating arthritis based on Endo180 activating use, comprising the Endo180 activating agent according to claim 1. Characterized by containing one or more components selected from collagen production promoters, anti-inflammatory agents, blood flow promoters, collagen, proteins, peptides, amino acids, black ginger, glucosamine, N-acetylglucosamine, and chondroitin. The composition for preventing/ameliorating arthritis according to claim 5. 7. A food or drink for preventing/improving arthritis based on Endo180 activation use, comprising the composition for preventing/improving arthritis according to claim 5 or 6.