Composition for promoting hyaluronic acid production

Abstract

To provide a composition for promoting hyaluronic acid production.SOLUTION: A composition for promoting hyaluronic acid production comprising at least one selected from the group consisting of (A) at least one selected from the group consisting of acetic acid and propionic acid or a salt thereof, (B) a prebiotic agent for bacteria that produce component (A), and (C) a probiotic agent for bacteria that produce component (A).SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a composition for promoting hyaluronic acid production. [Background technology]

[0002] Research into aging has been progressing in recent years. While aging is a major factor in skin aging from a macroscopic perspective, other direct factors involved in skin aging include dryness, oxidation, and sunlight (ultraviolet rays). Specific phenomena that are known to contribute to skin aging include a decrease in mucopolysaccharides such as hyaluronic acid, cross-linking of collagen, and cell damage caused by ultraviolet rays.

[0003] Hyaluronic acid, in particular, has many functions, including retaining moisture in intercellular spaces, retaining cells by forming a jelly-like matrix within tissues, maintaining tissue lubrication and flexibility, resisting external forces such as mechanical damage, and preventing bacterial infection (see Non-Patent Document 1). For example, it is said that the amount of hyaluronic acid in the skin decreases with age, resulting in the appearance of skin aging such as fine wrinkles and dryness. Therefore, many cosmetics containing hyaluronic acid or collagen have been proposed as agents for improving such aging skin. However, these conventional cosmetics only exert a moisturizing effect on the skin surface and are not capable of essentially improving aging skin. Cosmetics containing various vitamins and herbal medicines as skin cell activators have also been proposed, but these still do not currently have the ability to improve or treat aging skin.

[0004] Furthermore, hyaluronic acid contained in synovial fluid coats the surface of articular cartilage, contributing to the smooth functioning of joints. While the concentration of hyaluronic acid in normal human synovial fluid is approximately 2.3 mg / ml, in patients with rheumatoid arthritis, the concentration drops to approximately 1.2 mg / ml, and the viscosity of the synovial fluid also drops significantly (see Non-Patent Document 2). It is also known that hyaluronic acid content decreases in septic arthritis, gouty arthritis, and other conditions, similar to rheumatoid arthritis (see Non-Patent Document 3). Increasing the amount of hyaluronic acid in synovial fluid is considered to improve lubrication, coat and protect articular cartilage, suppress pain, and improve or normalize pathological synovial fluid in these diseases. For example, it has been reported that joint injection therapy with sodium hyaluronate in patients with rheumatoid arthritis results in the above-mentioned improvements (see Non-Patent Document 4). Similarly, joint injection therapy with hyaluronic acid has also been reported to have the same ameliorative effects in patients with traumatic arthritis, osteoarthritis, and degenerative arthritis. (See Non-Patent Document 5).

[0005] In this way, methods such as applying cosmetics containing hyaluronic acid to the skin or injecting hyaluronic acid directly into joints have been used, but even if hyaluronic acid is given externally, it does not fundamentally improve function and sufficient effects cannot be expected. In particular, hyaluronic acid has the problem that it is hardly absorbed by the skin.

[0006] For this reason, rather than simply supplying hyaluronic acid itself from the outside, there is a desire to develop a substance that utilizes the self-healing ability that humans naturally possess and promotes the hyaluronic acid production ability of human skin fibroblasts themselves, thereby fundamentally improving the body's functions. Among these, various plant-derived hyaluronic acid production promoters have been developed in the hope of being safe and causing mild irritation to the skin (see, for example, Patent Documents 1 to 4).

[0007] In addition, it is known that during the healing process after a burn, the amount of hyaluronic acid in the granulation tissue increases significantly from the early stage when granulation tissue grows from below the necrotic tissue until the entire tissue is replaced by granulation tissue (see Non-Patent Document 6), and hyaluronic acid production promoters are also expected to be used as early treatments for burns. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] Japanese Patent Application Publication No. 11-209261 [Patent Document 2] Japanese Patent Application Laid-Open No. 2007-84448 [Patent Document 3] Japanese Patent Application Laid-Open No. 2003-55244 [Patent Document 4] Japanese Patent Application Laid-Open No. 2012-56919 [Non-patent literature]

[0009] [Non-Patent Document 1] "Bio Industry", vol.8, p.346(1991) [Non-patent document 2] "Arthritis Rheumatism", vol.10, p.357(1967) [Non-patent document 3] "Bonding Composition" (Kanbara Publishing), 481 pages, 1984 [Non-patent document 4] "Inflammation" (Japanese Society of Inflammation), Vol. 11, p. 16, 1991 [Non-patent document 5] "Connective Tissue and Disease" (Kodansha), 246 pages, 1980 [Non-patent document 6] "Connective Tissue and Disease" (Kodansha), 153 pages, 1980 Summary of the Invention

[0010] The present invention provides a composition for promoting hyaluronic acid production.

[0011] The present inventors have surprisingly found that a composition comprising specific components can promote the production of hyaluronic acid, and the present invention is based on these findings.

[0012] According to the present invention, the following inventions are provided. (1) (A) one or more selected from the group consisting of acetic acid and propionic acid or salts thereof; (B) a prebiotic agent for bacteria that produce (A), and (C) A probiotic agent for bacteria that produces (A). A composition for promoting hyaluronic acid production, comprising one or more selected from the group consisting of: (2) (A) one or more selected from the group consisting of acetic acid and propionic acid or salts thereof; (B) a prebiotic agent for bacteria that produce (A), and (C) A probiotic agent for bacteria that produces (A). A composition for improving skin viscoelasticity, comprising one or more selected from the group consisting of: (3) The composition described in (2) for promoting hyaluronic acid production. (4) The composition according to any one of (1) to (3), wherein (B) comprises or consists of one or more selected from the group consisting of fructooligosaccharides, galactooligosaccharides, indigestible dextrin, α-cyclodextrin, dextran, isomaltodextrin, inulin, fucose, and plants of the Brassicaceae family. (5) The composition according to any one of (1) to (4), wherein (C) comprises or consists of one or more bacteria selected from the group consisting of the genera Akkermansia, Anaerostipes, Anaerobutyricum, Faecalibacterium, Roseburia, Coprobacter, Bacteroidetes, Clostridium, Propionibacterium, and Veillonella. (6) The composition according to any one of (1) and (3) to (5), which is a composition for promoting hyaluronic acid production in fibroblasts. (7) The composition according to any one of (1) to (6), which is a food composition.

[0013] According to the present invention, a composition for promoting hyaluronic acid production is provided. Furthermore, according to the present invention, it is also possible to provide a composition for improving the viscoelasticity of the skin. [Brief explanation of the drawings]

[0014] [Figure 1] FIG. 1 shows the results of evaluating the effect of each component on the expression of mRNA of the COL1A1 gene in human fibroblasts. [Figure 2] FIG. 2 shows the results of evaluating the effect of each component on the expression of mRNA of the COL3A1 gene in human fibroblasts. [Figure 3] FIG. 3 shows the results of evaluating the effect of each component on the expression of mRNA of the COL7A1 gene in human fibroblasts. [Figure 4] FIG. 4 shows the results of evaluating the effect of each component on the expression of mRNA of the HAS2 gene in human fibroblasts. [Figure 5]FIG. 5 is a graph showing the correlation between the skin viscoelasticity (R2) on the subject's arm and the amount of propionic acid in the feces. [Figure 6] FIG. 6 is a graph showing the correlation between the viscoelasticity (R2) of the skin on the face of the subjects and the amount of propionic acid in the feces. [Figure 7] FIG. 7 is a graph showing the correlation between skin viscoelasticity (R7) on the arm of a subject and the amount of propionic acid in the feces. [Figure 8] FIG. 8 shows the correlation between skin viscoelasticity (R7) on the subject's arm and the amount of acetic acid in the feces. Specific Description of the Invention

[0015] According to one embodiment of the present invention, there is provided a composition for promoting hyaluronic acid production, which comprises (A) one or more selected from the group consisting of acetic acid and propionic acid or a salt thereof, (B) a prebiotic agent for bacteria that produce (A), and (C) one or more selected from the group consisting of a probiotic agent for bacteria that produce (A). Such a composition has an excellent hyaluronic acid production-promoting effect and can be used for preventing skin aging, maintaining skin firmness and elasticity, preventing and treating arthritis, and the like, and for the early treatment of burns.

[0016] In particular, the present inventors have shown that there is a correlation between the amount of acetic acid or propionic acid in a subject's feces and skin viscoelasticity (see the Reference Examples and Figures 5-8 described below). Therefore, according to another aspect of the present invention, a composition for improving skin viscoelasticity is provided, which comprises (A) one or more selected from the group consisting of acetic acid and propionic acid or a salt thereof, (B) a prebiotic agent for bacteria that produce (A), and (C) one or more selected from the group consisting of a probiotic agent for bacteria that produce (A). The improvement of skin viscoelasticity by this composition may be achieved by promoting hyaluronic acid production, or may be achieved without promoting hyaluronic acid production.

[0017] In general, the health state of skin is evaluated using various indices. For example, the presence or absence of a feeling of "firmness" of the skin is considered one of the indices for evaluating the health state and degree of aging. "Firmness" can be divided into two types: firmness derived from the stratum corneum / epidermis, and firmness derived from the dermis. In particular, firmness derived from the dermis is 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. In this invention, skin viscoelasticity refers to such a physical property of the skin.

[0018] The evaluation of skin viscoelasticity in the present invention is not particularly limited to specific methods, but may be based on the results of measurements using, for example, a CUTOMETER (trade name) (manufactured by Courage+Khazaka Electronic GmbH). This device draws the skin surface into a probe opening under negative pressure, measures the length of the skin drawn into the opening with a prism, then releases the suction and similarly measures the displacement length (return) when released, and uses these measurement results as parameters to determine skin viscoelasticity (see the product's instruction manual).

[0019] The evaluation of skin viscoelasticity in the present invention is not particularly limited, and may be based on parameters commonly used as indicators of skin viscoelasticity. Examples of such parameters include R2, the final return rate of the skin when suction is released relative to the maximum length stretched during suction (R2 = Ua / Uf), R5, the instantaneous return rate of the skin when suction is released relative to the instantaneous length stretched during suction (R5 = Ur / Ue), and R7, the instantaneous (instantaneous) contraction rate of the skin when suction is released relative to the length stretched during suction (R7 = Ur / Uf). In the formula, Uf, Ur, Ue, and Ua represent Uf: maximum skin stretch during suction, Ur: instantaneous (instantaneous: 0.1 seconds) skin return after suction is released, Ue: instantaneous (instantaneous: 0.1 seconds) skin stretch due to suction, and Ua: final return rate of the skin after suction is released, respectively. Skin viscoelasticity in the present invention may be evaluated based on one or more parameters selected from the group consisting of R2, R5, and R7.

[0020] In the present invention, the site for evaluating skin viscoelasticity may be any part of the skin, and examples include, but are not limited to, the skin of the face (e.g., forehead, cheek, etc.), forearm, upper arm, back, stomach, etc. Evaluation of skin viscoelasticity may be performed in a beauty salon, a cosmetics store, an esthetic salon, etc., and may be performed by a non-diagnostic method or a diagnostic aid method.

[0021] According to one embodiment of the present invention, the composition of the present invention comprises (A) one or more selected from the group consisting of acetic acid and propionic acid or a salt thereof.

[0022] The acetate salt in the present invention is not particularly limited, and examples thereof include sodium acetate and potassium acetate.

[0023] The propionate salt in the present invention is not particularly limited, and examples thereof include sodium propionate and potassium propionate.

[0024] According to one preferred embodiment of the present invention, the composition of the present invention comprises acetic acid, sodium acetate, propionic acid, or sodium propionate.

[0025] According to another embodiment of the present invention, the composition of the present invention comprises (B) a prebiotic agent for bacteria that produce (A).

[0026] In the present invention, (B) is not particularly limited as long as it is used to induce the growth or activity of the bacteria that produce (A), and examples thereof include growth promoters and metabolic activators for the bacteria that produce (A).

[0027] In the present invention, (B) may comprise or consist of a component or food material that has the effect of inducing the growth or activity of the bacterium that produces (A). Such components are preferably fructooligosaccharides, galactooligosaccharides, indigestible dextrin, α-cyclodextrin, dextran, isomaltodextrin, inulin, or fucose. Furthermore, such food materials are preferably plants of the Brassicaceae family, more preferably broccoli sprouts.

[0028] According to one preferred embodiment of the present invention, the composition of the present invention comprises (B) a prebiotic agent for bacteria that produce (A), wherein (B) comprises or consists of one or more selected from the group consisting of fructooligosaccharides, galactooligosaccharides, indigestible dextrin, α-cyclodextrin, dextran, isomaltodextrin, inulin, fucose, and plants of the Brassicaceae family.

[0029] According to another embodiment of the present invention, the composition of the present invention comprises (C) a probiotic agent for bacteria producing (A).

[0030] In the present invention, (C) comprises or consists of a bacterium that produces (A).

[0031] In the present invention, the bacterium that produces (A) is not particularly limited.

[0032] According to one preferred embodiment of the present invention, the bacterium producing (A) of the present invention is of the genus Akkermansia, more preferably Akkermansia muciniphila.

[0033] According to another preferred embodiment of the present invention, the bacterium producing (A) of the present invention is of the genus Anaerostipes, more preferably Anaerostipes caccae.

[0034] According to another preferred embodiment of the present invention, the bacterium producing (A) of the present invention is the genus Anaerobutyricum (Eubacterium), more preferably Anaerobutyricum hallii (Eubacterium hallii).

[0035] According to another preferred embodiment of the present invention, the bacterium producing (A) of the present invention is of the genus Faecalibacterium, more preferably Faecalibacterium prausnitzii.

[0036] According to another preferred embodiment of the present invention, the bacterium producing (A) of the present invention is of the genus Roseburia, more preferably Roseburia intestinalis.

[0037] According to another preferred embodiment of the present invention, the bacterium producing (A) of the present invention is a Coprobacter genus.

[0038] According to another preferred embodiment of the present invention, the bacterium producing (A) of the present invention is a Bacteroidetes genus.

[0039] According to another preferred embodiment of the present invention, the bacterium producing (A) of the present invention is of the genus Clostridium, more preferably Clostridium cluster IX.

[0040] According to another preferred embodiment of the present invention, the bacterium producing (A) of the present invention is of the genus Propionibacterium.

[0041] According to another preferred embodiment of the present invention, the bacterium producing (A) of the present invention is a Veillonella genus.

[0042] According to another preferred embodiment of the present invention, the composition of the present invention comprises (C) a probiotic agent for bacteria that produce (A), wherein (C) comprises or consists of one or more bacteria selected from the group consisting of the genera Akkermansia, Anaerostipes, Anaerobutyricum, Faecalibacterium, Roseburia, Coprobacter, Bacteroidetes, Clostridium, Propionibacterium, and Veillonella.

[0043] The compositions of the present invention may contain (A), (B), and (C) alone or in combination.

[0044] The composition of the present invention may be applied either in vivo or in vitro.

[0045] The cells to which the composition of the present invention is applied are not limited, and may be, for example, cells in a living body, cells obtained from a living body, passaged cells thereof, or established cell lines, and these cells may be differentiated and used as needed. The cells in the present invention are preferably fibroblasts.

[0046] When the composition of the present invention is applied to a subject, examples of the subject to which it can be applied include, but are not limited to, humans, primates including chimpanzees, pet animals such as dogs and cats, livestock animals such as cows, horses, sheep and goats, and mammals such as rodents including mice and rats, and preferably humans.

[0047] The composition of the present invention may be formulated as a food composition, although it is not particularly limited thereto. When the composition of the present invention is formulated as a food composition, known ingredients (additives) formulated in food compositions may be further formulated. Examples of such ingredients include sweeteners, high-sugar sweeteners (aspartame, sucralose, glycyrrhizin, saccharin, stevia, dulcin, trichlorosucrose, thaumatin, acesulfame potassium, etc.), flavorings, colorings, acidulants, antioxidants, emulsifiers, preservatives, stabilizers, etc.

[0048] The food composition of the present invention may be a health functional food such as a nutrient functional food, a food for specified health uses, or a food with functional claims, or may be manufactured as a therapeutic food (i.e., a food that serves a therapeutic purpose, or a food prepared based on a menu prepared by a nutritionist or the like in accordance with a doctor's dietary prescription), a dietary therapy food, a nursing care food, etc. [Example]

[0049] The present invention will be described in detail based on the following examples, but the present invention is not limited to these examples. Contents are expressed in mass % unless otherwise specified.

[0050] Test Example 1: Evaluation of the effect of each component on the production of hyaluronic acid, etc. in human fibroblasts Human fibroblasts (product name: "Skin fibroblasts", manufactured by Kurabo Industries, Ltd.) were dispersed in Dulbecco's Modification of Eagle's Medium (DMEM) containing 10% fetal bovine serum (FBS) at a density of 2.5 × 10 5The concentration of 1000 cells / mL was adjusted. 2 mL of this solution was seeded into each well of a 6-well plate (Corning Costar 3516). The human fibroblasts seeded in this 6-well plate were cultured in an incubator set at 37°C and 5% CO2. After 48 hours, the medium was removed and replaced with DMEM containing 1% antibiotic (product name: Antibiotic-Antimycotic Mixed Stock Solution (100x) (Stabilized), Nacalai Tesque), and the culture was continued for another 24 hours.

[0051] After 24 hours, the medium was removed and replaced with DMEM containing various components as listed in Table 1 below, and the cells were cultured for an additional 18 hours to prepare cells cultured under four different conditions (Examples 1 and 2 and Comparative Examples 1 and 2).

[0052] [Table 1]

[0053] The mRNA expression of COL1A1, COL3A1, COL7A1, and HAS2 genes was evaluated in each of the four cell types. First, mRNA for COL1A1, COL3A1, COL7A1, and HAS2 genes was extracted and purified from the four cell types using the RNeasy Mini Kit (QIAGEN) according to the provided protocol. Using the extracted RNA, semi-quantitative PCR of COL1A1, COL3A1, COL7A1, and HAS2 genes was performed (Light Cycler 480; Roche) using the TaqMan RNA-to-CT 1-step Kit (Thermo Fisher Scientific) according to the provided protocol. The expression levels of each gene were normalized using β-actin as an internal standard. The primers used were as follows (all from Thermo Fisher Scientific): COL1A1:Hs00164004_m1 COL3A1:Hs00943809_m1 COL7A1:Hs00164310_m1 HAS2:Hs00193435_m1 β-actin: Hs01060665_g1

[0054] The ratio of the expression level of COL1A1 mRNA obtained from each of the cell groups of Comparative Example 2, Example 1, and Example 2 to the expression level of COL1A1 mRNA obtained from the control (Comparative Example 1) is shown in Figure 1. Furthermore, the ratio of the expression level of COL3A1 mRNA obtained from each of the cell groups of Comparative Example 2, Example 1, and Example 2 to the expression level of COL3A1 mRNA obtained from the control (Comparative Example 1) is shown in Figure 2. Furthermore, the ratio of the expression level of COL7A1 mRNA obtained from each of the cell groups of Comparative Example 2, Example 1, and Example 2 to the expression level of COL7A1 mRNA obtained from the control (Comparative Example 1) is shown in Figure 3. Additionally, the ratio of the expression level of HAS2 mRNA obtained from each of the cell groups of Comparative Example 2, Example 1, and Example 2 to the expression level of HAS2 mRNA obtained from the control (Comparative Example 1) is shown in Figure 4.

[0055] These results indicated that IL-22 had little effect on the mRNA expression of the COL1A1, COL3A1, COL7A1, and HAS2 genes, whereas sodium acetate and sodium propionate significantly increased the mRNA expression of the HAS2 gene.

[0056] Reference example In this example, the correlation between skin viscoelasticity and the amount of short-chain fatty acids in feces was shown as follows.

[0057] Measurement of skin viscoelasticity The skin viscoelasticity (R2 or R7 on the face or arm) of the human subjects was measured by the following method.

[0058] Skin viscoelasticity was measured on the cheeks and inner upper arms of 20 healthy women aged 20 to 50 years using a measuring device (Cutometer MPA580 (Courage+Khazaka Electronic Measurements were performed using a suction device (manufactured by GE Healthcare GmbH) according to the following procedure. First, the subjects' face and inner upper arms, which were the measurement sites, were washed and then allowed to acclimate for 30 minutes in a constant temperature and humidity chamber (temperature 21±2°C, humidity 45%±5%) while remaining still. Next, measurements were performed three times using a φ2mm probe for 60 seconds, with 400mb suction pressure, and a 1-second suction followed by a 1-second pause, repeated 30 times. The average value was used as the measured value. Measurements on the subject's cheek were performed using an L-shaped scale at the intersection of the underside of the nose and the corner of the eye, approximately 3 cm below and 3 cm to the left of that point. Measurements on the subject's inner upper arm were performed starting from the sharp bone on the inner elbow, 1.5 cm inward and 5 cm toward the shoulder, and then 1.5 cm inward and 4.5 cm toward the shoulder from that point.

[0059] Measurement of short-chain fatty acids in feces The amounts of short-chain fatty acids (acetic acid and propionic acid) in the feces of the human subjects were measured by the following method.

[0060] Feces from 20 healthy women aged 20 to 50 were collected using a stool collection kit (product name: Feces Collection Kit (manufactured by Techno Suruga Lab Co., Ltd.)), and the amount of short-chain fatty acids (μmol / g) in the feces was quantified. The quantification was outsourced to a specialized third-party institution.

[0061] statistical processing We investigated the correlation between skin viscoelasticity and the amount of short-chain fatty acids in feces. When the correlation coefficient was calculated by plotting the skin viscoelasticity and the amount of short-chain fatty acids in feces on a scatter plot, it was found that there was a significant correlation or a correlation that tended to be significant between skin viscoelasticity and the amount of short-chain fatty acids in feces (Figure 5: p=0.05323, Figure 6: p=0.04442, Figure 7: p=0.05711, Figure 8: p=0.08162).

Claims

1. (A) one or more selected from the group consisting of acetic acid and propionic acid or salts thereof; (B) a prebiotic agent for bacteria that produce (A), and (C) A probiotic agent for bacteria producing (A). A composition for promoting hyaluronic acid production, comprising one or more selected from the group consisting of:

2. (A) one or more selected from the group consisting of acetic acid and propionic acid or salts thereof; (B) a prebiotic agent for bacteria that produce (A), and (C) A probiotic agent for bacteria producing (A). A composition for improving skin viscoelasticity, comprising one or more selected from the group consisting of:

3. 3. The composition according to claim 1 or 2, wherein (B) comprises or consists of one or more selected from the group consisting of fructooligosaccharides, galactooligosaccharides, indigestible dextrin, α-cyclodextrin, dextran, isomaltodextrin, inulin, fucose, and plants of the Brassicaceae family.

4. 3. The composition of claim 1 or 2, wherein (C) comprises or consists of one or more bacteria selected from the group consisting of the genera Akkermansia, Anaerostipes, Anaerobutyricum, Faecalibacterium, Roseburia, Coprobacter, Bacteroidetes, Clostridium, Propionibacterium, and Veillonella.

5. The composition according to claim 1, which is a composition for promoting hyaluronic acid production in fibroblasts.

6. 3. The composition of claim 1 or 2, which is a food composition.

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