Agents for promoting elastin production

Abstract

To discover a material having an action of promoting elastin production and provide an agent and the like for promoting elastin production.SOLUTION: An agent for promoting elastin production contains proteoglycan. An oral composition contains the agent.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an agent for promoting elastin production, for example, for use in humans, etc. [Background technology]

[0002] As the first line of defense against direct external environmental influences, the skin plays an important role in maintaining the internal environment of the body. While skin function never completely ceases, it gradually declines with age, exposure to ultraviolet light, and exposure to chemicals, resulting in the appearance of wrinkles, age spots, dullness, sagging skin, and other signs of aging. For example, the presence or absence of skin firmness is considered one indicator of health and aging. "Firmness" can be divided into two categories: firmness derived from the stratum corneum / epidermis and firmness derived from the dermis. Dermal firmness, in particular, refers to a state in which the skin has an elasticity that pushes back when pressed with a finger and quickly returns to its original state when the finger is released; physically, this is also known as viscoelasticity. While the detailed mechanism behind the decline in skin viscoelasticity is not fully understood, thinning of the skin is thought to be one of the contributing factors (Patent Document 1).

[0003] Elastin is a fiber that provides elasticity to skin tissue. Elastin production decreases with aging and other factors, and its degradation and denaturation are accelerated by ultraviolet light. A decrease in normal elastin reduces skin elasticity, which can lead to wrinkles and sagging. Therefore, if elastin production can be promoted, it is believed that wrinkles and sagging will be less likely to occur, and that symptoms of skin aging, such as loss of firmness and loss of elasticity, can be prevented and / or improved (Patent Document 1). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent 7144878 Summary of the Invention [Problem to be solved by the invention]

[0005] The problem to be solved by the present invention is to find naturally occurring substances (natural products) that have an elastin production-promoting effect, and to provide agents for promoting elastin production. [Means for solving the problem]

[0006] Therefore, the inventors of the present invention conducted extensive research to find substances that have the activity of promoting elastin production, and as a result discovered that elastin production can be achieved by adding proteoglycan to certain cells, etc. (for example, without adding elastin), and thus completed the present invention. [Item 1] An agent for promoting elastin production, containing proteoglycan. [Item 2] The agent according to [Item 1] for promoting extracellular matrix production. [Item 3] The agent described in [Item 2] for promoting collagen production and / or hyaluronic acid production. [Item 4] The agent according to [Item 2] or [Item 3] for improving skin elasticity. [Item 5] An oral composition containing the agent according to any one of [Item 1] to [Item 4]. [Effects of the Invention]

[0007] The present invention can provide a new oral composition for promoting elastin production, etc. DETAILED DESCRIPTION OF THE INVENTION

[0008] Hereinafter, an embodiment of the present invention will be described.

[0009] (proteoglycan) Proteoglycans (PGs) are glycoproteins composed of core proteins to which sugar chains called glycosaminoglycans (GAGs), such as chondroitin sulfate and dermatan sulfate, are covalently bonded. PGs are widely distributed throughout the body, including skin and cartilage, as one of the major components of the extracellular matrix. GAG chains have a long, unbranched, linear structure. They are negatively charged due to the large number of sulfate and carboxyl groups they contain, and the electrical repulsion between them results in an extended shape. PGs can also retain large amounts of water due to the water affinity of their sugars. The numerous GAG chains contained in PG flexibly retain water like a sponge, while also providing cartilage-specific functions such as elasticity and shock resistance.

[0010] The core protein of PG has the property of binding to various molecules in the matrix. In the case of cartilage PG, it has a binding domain for hyaluronic acid and link protein at the N-terminus, and it binds to these substances and can also associate with the same molecule. The C-terminus contains a lectin-like domain, EGF-like domain, etc., and it binds to various other molecules. This property allows PG to build a structure that is suited to each tissue.

[0011] Salmon nasal cartilage-derived PG is PG extracted from salmon nasal cartilage. Here, the salmon is, for example, a fish belonging to the genus Oncorhynchus, and preferably, salmon with the scientific name "Oncorhynchus keta" is selected from the viewpoint of efficiently regulating immune responses. The proteoglycan contained in the agent or composition of this embodiment is prepared, for example, by the method described in the publication (Japanese Patent No. 6317053). Furthermore, the content of proteoglycan in a composition (1 g) containing the composition of this embodiment is preferably 0.1 μg / g or more, more preferably 100 μg / g or more, and even more preferably 1 mg / g or more, from the viewpoint of effectively exerting elastin production, for example.

[0012] An example of a commercially available PG product is Proteoglycan F (manufactured by Ichimaru Pharcos).

[0013] (Promotes elastin production) Increasing the amount of elastin in the body (promoting elastin production) can increase the strength and elasticity of the dermis, ligaments, tendons, blood vessel walls, bones, cartilage, etc. Agents for elastin production are thought to be used to prevent and / or improve various disorders associated with elastin reduction (e.g., for treating or preventing vascular diseases such as arteriosclerosis caused by a lack of normal elastin), and to prevent and / or improve cosmetic problems (e.g., for preventing and / or improving wrinkles or sagging skin caused by elastin reduction due to aging, ultraviolet light, active oxygen, stress, etc., or for preventing and / or improving loss of skin elasticity or firmness).

[0014] (extracellular matrix) The extracellular matrix (ECM) is a complex of carbohydrates and proteins present in the basement membrane that lines tissues and in the intercellular spaces. Examples of structures formed by the ECM include the basement membrane, glial scar, and perineuronal net.

[0015] (collagen) Collagen is a major fibrous component that supports the structure of biological tissues such as skin, bones, and tendons, and is an essential protein found in large amounts in the body of animals.

[0016] (Hyaluronic acid) Hyaluronic acid is the polysaccharide that has the structure that the disaccharide unit of glucuronic acid and N-acetylglucosamine is connected.In addition, the salt of hyaluronic acid is not particularly limited, and it can be any salt that can be food or pharmacy acceptable, for example, sodium salt, potassium salt, calcium salt, zinc salt, magnesium salt, ammonium salt etc.The molecular weight of hyaluronic acid is not particularly limited.

[0017] (Promotes hyaluronic acid production) In the present invention, "promoting hyaluronic acid production" is not limited by its mechanism, as long as it primarily promotes the synthesis of hyaluronic acid in keratinocytes in the epidermis (particularly the cornified layer). Promoting hyaluronic acid production is preferably achieved, for example, by promoting the expression of genes encoding mammalian hyaluronic acid synthases HAS2 and HAS3. In the present invention, the hyaluronic acid production promoters of the present invention can be used not only as hyaluronic acid production promoters, but also as hyaluronic acid synthase expression promoters (e.g., hyaluronic acid gene expression promoters), particularly as promoters of HAS2 gene or HAS3 gene expression. The hyaluronic acid production promoters of the present invention can prevent functional decline in various disorders, diseases, and illnesses associated with a decrease in hyaluronic acid in the body, particularly in the skin. For example, they can promote skin moisture, improving skin firmness and elasticity while providing moisture, and can prevent or improve skin problems such as rough skin, wrinkles, and dryness, as well as joint disorders and joint damage.

[0018] (Improves skin elasticity) "Improving skin elasticity" not only means "suppressing the loss of skin elasticity (preventing the loss of skin elasticity, etc.)," ​​but also includes, for example, "improving the symptoms of loss of skin elasticity (increasing skin thickness, etc.) by administering a specified composition (e.g., orally) after a loss of skin elasticity has occurred."

[0019] (Oral composition) The oral composition according to the present invention is, for example, a food or drink (including functional foods, foods for specified health uses, supplements, etc.), a pharmaceutical product, or the like.

[0020] For example, when the oral composition is a food or beverage, the food or beverage may take the form of various foods and beverages such as breads, cakes, noodles, confectioneries, jellies, frozen foods, ice cream, dairy products, beverages, etc., as well as forms similar to those of the oral administration preparations described above (tablets, capsules, syrups, etc.) Foods in various forms can be prepared by using the active ingredient of the present invention alone or in appropriate combination with other food ingredients, solvents, softeners, oils, emulsifiers, preservatives, flavorings, stabilizers, colorants, antioxidants, moisturizers, thickeners, etc.

[0021] For example, when the oral composition is a pharmaceutical, the pharmaceutical is generally easy to formulate a convenient daily dosage regimen that can be adjusted according to the degree of pain, and the pharmaceutical form may be, for example, a solid form or a liquid form. Such solid forms include, for example, powders, tablets, pills, capsules, cachets, lozenges, suppositories, and dispersible granules. For example, in powders, the carrier is generally a finely divided solid that is mixed with a finely divided active ingredient. For example, in tablets, the active ingredient is generally mixed with a carrier having the necessary binding capacity in an appropriate ratio and compacted into the desired shape and size. Suitable carriers may include, but are not limited to, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low-melting wax, cocoa butter, etc. The pharmaceutical may also contain ingredients such as excipients, stabilizers, preservatives, binders, disintegrants, hydrocarbons, fatty acids, alcohols, esters, pH adjusters, and preservatives as needed, within the range that does not impair the desired effects.

[0022] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples. In the following examples, the unit % used to indicate the amount of each component added means % by mass. [Example]

[0023] Examples of the present invention will be described below. The samples used in the experiments are as follows: Proteoglycan: Proteoglycan F (proteoglycan content 20.0% or more, Ichimaru Falcos). In the following experiments, proteoglycan F was used to match the specified amount of proteoglycan. Normal human adult skin fibroblasts: fibroblasts derived from neonatal foreskin skin, Kurabo, KF-4009 D-MEM medium: Fujifilm Wako Pure Chemical Industries, 041-29775, low glucose (containing L-glutamine and phenol red)

[0024] [Experiment 1]: Evaluation of elastin production promoting ability This evaluation was carried out by confirming whether the addition of proteoglycan promotes elastin production in normal adult human skin fibroblasts.

[0025] (Experimental Method) 1. Cell Preparation D-MEM medium (Fujifilm Wako Pure Chemical Industries, product number 187-02705) containing 5% FBS and normal human adult dermal fibroblasts were added to the medium. After the addition, the medium was replaced with fresh medium and cultured under conditions of 5% CO2 and 37°C until the cells became 70-80% confluent. After the culture, the cells were washed with PBS and then cultured in "TrypLE" TM Normal human adult skin fibroblasts were detached from the flask using Select Enzyme (1x), no phenol red (Gibco, product number: 12563011) and subcultured.

[0026] After the subculture, normal human adult skin fibroblasts were cultured in a 24-well cell culture plate (containing D-MEM medium containing 5% FBS) at 4 × 10 4 The cells were seeded at 100 cells / well. The plate was cultured until 100% confluent, with the medium being replaced with fresh medium. After this culture, the medium in the plate was replaced with D-MEM medium containing 0.1% FBS. After this replacement, the plate was cultured for 24 hours under conditions of 5% CO2 and 37°C.

[0027] 2. Adding Sample to Cells After the 24-hour culture, the medium was replaced with the medium (culture conditions) shown in Table 1, and the cells were cultured under conditions of 5% CO2 and 37°C for 24 hours.

[0028] 3. Culture medium collection and ELISA evaluation After the culture in step 2, the supernatant of the culture medium was collected, and the amount of elastin in the supernatant was evaluated using a Human Elastin ELISA kit (CUSABIO, CSB-E09338h).

[0029] In Experiment 1, the control group was a group to which the proteoglycan was not added (0 μg / mL). This experiment was conducted with three experimental groups: the control group, a group to which 10 μg / mL of proteoglycan was added (PG10), and a group to which 100 μg / mL of proteoglycan was added (PG100). Three samples (n=3) were prepared for each of the three experimental groups, and Experiment 1 was conducted. The experimental results (elastin production amount) of Experiment 1 are as follows:

[0030] [Table 1]

[0031] [Experiment 2]: Evaluation of collagen production promoting ability This evaluation was carried out by confirming whether the addition of proteoglycan promotes collagen production in normal adult human skin fibroblasts.

[0032] (Experimental Method) 1. Cell Preparation D-MEM medium (Fujifilm Wako Pure Chemical Industries, product number 187-02705) containing 5% FBS and normal human adult dermal fibroblasts were added to the medium. After the addition, the medium was replaced with fresh medium and cultured under conditions of 5% CO2 and 37°C until the cells became 70-80% confluent. After the culture, the cells were washed with PBS and then cultured in "TrypLE" TMNormal human adult skin fibroblasts were detached from the flask using Select Enzyme (1x), no phenol red (Gibco, product number: 12563011) and subcultured.

[0033] After the subculture, normal human adult skin fibroblasts were cultured in a 24-well cell culture plate (containing D-MEM medium containing 5% FBS) at 4 × 10 4 The cells were seeded at 100 cells / well. The plate was cultured until 100% confluent, with the medium being replaced with fresh medium. After this culture, the medium in the plate was replaced with D-MEM medium containing 0.1% FBS. After this replacement, the plate was cultured for 24 hours under conditions of 5% CO2 and 37°C.

[0034] 2. Adding Sample to Cells After the 24-hour culture, the medium was replaced with the medium (culture conditions) shown in Table 1, and the cells were cultured under conditions of 5% CO2 and 37°C for 24 hours.

[0035] 3. Culture medium collection and ELISA evaluation After the culture in step 2, the supernatant of the culture medium was collected, and the concentration of type III collagen in the supernatant was evaluated using a Procollagen type I C-peptide (PIP) EIA Kit (MK101, Cosmo Bio).

[0036] In Experiment 2, the control group was a group to which no proteoglycan was added (0 μg / mL). This experiment was conducted with three experimental groups: the control group, a group to which 10 μg / mL of proteoglycan was added (PG10), and a group to which 100 μg / mL of proteoglycan was added (PG100). Three samples (n=3) were prepared for each of the three experimental groups, and Experiment 1 was conducted using these samples. The experimental results (elastin production amount) of Experiment 2 are as follows:

[0037] [Table 2]

[0038] [Experiment 3]: Evaluation of skin condition improvement effects in human monitor tests This monitoring study was conducted from February 2021 to April 2021 on 90 healthy men and women (30 people per group, 3 groups) aged 30 to 60 (average age 48.4 years) who were aware of sagging, dry, and / or rough skin.

[0039] (Test Method) The groups shown in Table 3 were instructed to consume one bottle of the test drink once a day between meals.

[0040] [Table 3]

[0041] (measuring skin viscoelasticity) The viscoelasticity of the subjects' skin (left cheek) was measured using a viscoelasticity measuring device (Cutometer, MPA580), and the relative values ​​were calculated compared to the values ​​before intake for each of the R5 and R7 groups, which were set at 100. The elasticity parameters (R5, R7) are as follows:

[0042] R5: Net elasticity. Ratio of UR to UE. UR is the height of elastic elongation at relaxation. UE is the height of elastic elongation when sucked into the probe. R7: Recovery elasticity, the ratio of UR to UF, the instantaneous return rate at which the elastic properties appear for the lifted skin.

[0043] The measurement results of the relative values ​​of R5 and R7 are shown in Table 4. Compared to the placebo group, Groups 1 and 2 had better viscoelasticity.

[0044] [Table 4]

[0045] (Measurement of transepidermal water loss) The transepidermal water loss of the subjects' skin (left cheek) was measured using a Tewameter (TM300, Courage-Khazaka), and the relative value was calculated compared to the value before intake for each group, which was set at 100. The measurement results are shown in Table 5. Compared to the placebo group, transepidermal water loss was lower in Groups 1 and 2.

[0046] [Table 5]

[0047] (Skin moisture measurement) The moisture content of the subjects' skin (left cheek) was measured using a Corneometer (CM825, Courage-Khazaka). The value before intake for each group was set at 100, and a relative value was calculated. The measurement results are shown in Table 6. Compared to the placebo group, Groups 1 and 2 had higher moisture content.

[0048] [Table 6]

[0049] The above has described the embodiments (including examples) of the present invention with reference to the drawings, but the specific configuration of the present invention is not limited to this, and even if there are design changes, etc., within the scope that does not deviate from the gist of the present invention, they are included in the present invention. [Industrial Applicability]

[0050] The present invention can provide an agent for promoting elastin production that contains proteoglycan.

Claims

1. An agent for promoting elastin production, containing proteoglycan extracted from salmon nasal cartilage with an acetic acid solution.

2. An agent for promoting elastin production and collagen production, comprising proteoglycan extracted from salmon nasal cartilage with an acetic acid solution. End

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