Agents for adjusting biological functions, epidermal metabolism promoters, fat accumulation inhibitors, fat decomposition promoters, adiponectin production promoters, functional foods, cosmetics, and methods for producing agents for adjusting biological functions

Abstract

A biological function regulating agent comprises a fermented collagen peptide, in which the fermented collagen peptide has at least one activity selected from the group consisting of an epidermal metabolism promoting activity, a fat accumulation inhibiting activity, a fat decomposition promoting activity and an activity to control the amount of an adipocytokine in a living body.

Description

Technical Field

[0001] The present invention relates to a biological function regulator, an epidermal metabolism promoter, a fat accumulation inhibitor, a fat decomposition promoter, an adiponectin production promoter, a functional food, a cosmetic, and a method for producing a biological function regulator.

Background Art

[0002] International Publication No. 2017 / 014149 (Patent Document 1), a paper by Woo et al. (Non-Patent Document 1), and a presentation by Kobayashi et al. (Non-Patent Document 2) report that collagen peptide has an anti-obesity effect. Japanese Patent Application Laid-Open No. 2018-023326 (Patent Document 2) discloses that collagen peptide was obtained by fermenting collagen with lactic acid bacteria.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Patent Document 2

Non-Patent Documents

[0004]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0005] The above Patent Document 2 does not disclose obtaining collagen peptides by fermenting any of collagen, gelatin, and gelatin degradation products with koji. Furthermore, the collagen peptides reported to have an anti-obesity effect in the above Patent Document 1, the above Non-Patent Document 1, and the above Non-Patent Document 2 are not collagen peptides obtained by fermenting any of collagen, gelatin, and gelatin degradation products with koji (hereinafter, also referred to as "fermented collagen peptides"). That is, the effects of fermented collagen peptides on the living body have not yet been elucidated.

[0006] In view of the above circumstances, an object of the present invention is to provide a biological function regulator, an epidermis metabolism promoter, a fat accumulation inhibitor, a lipolysis promoter, an adiponectin production promoter, a functional food, a cosmetic, and a method for producing a biological function regulator, which contain a fermented collagen peptide having at least one action selected from the group consisting of an epidermis metabolism promoting action, a fat accumulation inhibiting action, a lipolysis promoting action, and an action of regulating the amount of adipocytokines in the living body.

Means for Solving the Problems

[0007] The present inventors have found that fermented collagen peptides obtained by fermenting raw materials containing collagen such as the skin, skin, bone, cartilage, and tendon of tetrapods, and the bone, skin, and scale of fish, or any of collagen, gelatin, and gelatin degradation products with koji have at least one action selected from the group consisting of an epidermis metabolism promoting action, a fat accumulation inhibiting action, a lipolysis promoting action, and an action of regulating the amount of adipocytokines in the living body, and have completed the present invention.

[0008] That is, the present invention has the following features. [1] The biological function regulator according to the present invention contains fermented collagen peptide, and the fermented collagen peptide has at least one action selected from the group consisting of an action of promoting epidermal metabolism, an action of suppressing fat accumulation, an action of promoting fat decomposition, and an action of regulating the amount of adipocytokines in the living body. [2] The fermented collagen peptide preferably contains a collagen peptide and at least one first compound selected from the group consisting of isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional. [3] The fermented collagen peptide preferably contains a collagen peptide and at least three first compounds selected from the group consisting of isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional. [4] The epidermal metabolism promoter according to the present invention contains the above biological function regulator. [5] The fat accumulation inhibitor according to the present invention contains the above biological function regulator. [6] The adiponectin production promoter according to the present invention contains the above biological function regulator. [7] The fat decomposition promoter according to the present invention contains the above biological function regulator. [8] The functional food according to the present invention contains the above biological function regulator. [9] The cosmetic according to the present invention contains the above biological function regulator.

[10] The method for producing a biological function regulator according to the present invention is a method for producing a biological function regulator containing fermented collagen peptide, and includes a step of preparing koji containing Aspergillus and a collagen raw material, and a step of obtaining a biological function regulator containing the fermented collagen peptide by fermenting the collagen raw material with the koji. The bacterial species of the Aspergillus is a bacterial species belonging to the genus Aspergillus, and the collagen raw material is at least one selected from the group consisting of the following Group 1 to Group 6, collagen extracted from at least one selected from the group, gelatin obtained by treating the collagen, and at least any one of gelatin degradation products obtained by hydrolyzing the gelatin. Group 1: The group consisting of cowhide, skin, bone, cartilage, and tendon Group 2: a group consisting of pig skin, skin, bone, cartilage and tendon Group 3: a group consisting of sheep skin, skin, bone, cartilage and tendon Group 4: a group consisting of chicken skin, skin, bone, cartilage and tendon Group 5: a group consisting of ostrich skin, skin, bone, cartilage and tendon Group 6: a group consisting of fish bone, skin and scale 〔11〕 The biological function regulator according to the present invention contains a fermented collagen peptide produced by fermenting a collagen raw material with koji.

Advantages of the Invention

[0009] According to the above, a biological function regulator containing a fermented collagen peptide having at least one action selected from the group consisting of an epidermal metabolism promoting action, a fat accumulation inhibiting action, a fat decomposition promoting action and an action of regulating the amount of adipocytokines in vivo, an epidermal metabolism promoter, a fat accumulation inhibitor, a fat decomposition promoter, an adiponectin production promoter, a functional food, a cosmetic and a method for producing a biological function regulator can be provided.

Brief Description of the Drawings

[0010]

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Embodiments for Carrying Out the Invention

[0011] Hereinafter, embodiments according to the present invention (hereinafter also referred to as "the present embodiments") will be described in more detail. Here, in this specification, the notation in the form of "A to B" means the upper and lower limits of the range (that is, A or more and B or less). When there is no unit description for A and there is a unit description only for B, the unit of A and the unit of B are the same.

[0012] In this specification, "biological function regulator", "epidermal metabolism promoter", "fat accumulation inhibitor", "lipolysis promoter", "adiponectin production promoter", and "fermented collagen peptide" may be in a solid state such as powder, or may be in a liquid state such as an aqueous solution dissolved in water. Further, in this specification, "fermented collagen peptide" means a peptide mixture obtained by fermenting a collagen raw material described later with koji. In this specification, "fermentation" means the entire process in which beneficial organic substances are produced from raw materials by the activity of koji mold contained in koji, and is distinguished from "putrefaction" in which non-beneficial organic substances are produced from raw materials by the activity of microorganisms.

[0013] In this specification, the term "gelatin" may be used when referring to the substance name, gelatin gel, and gelatin solution, respectively. Also, for the term "collagen peptide", similar to the above gelatin, it may be used when referring to the substance name and collagen peptide solution, respectively.

[0014] In this specification, the "collagen raw material" may be used to refer collectively to at least one "itself" selected from the group consisting of the following Group 1 to Group 6, "collagen" extracted from at least one selected from the group consisting of the following Group 1 to Group 6, "gelatin" obtained by treating the above collagen using a known method such as hot water extraction, and "gelatin hydrolyzate" obtained by hydrolyzing the above gelatin. Further, the "hydrolysis" of the above gelatin includes all of hydrolysis using an acid, hydrolysis using a base, hydrolysis using an enzyme, and hydrolysis using heat. Group 1: The group consisting of cowhide, skin, bone, cartilage, and tendon Group 2: The group consisting of pigskin, skin, bone, cartilage, and tendon Group 3: The group consisting of sheepskin, skin, bone, cartilage, and tendon Group 4: The group consisting of chicken skin, skin, bone, cartilage, and tendon Group 5: The group consisting of ostrich skin, skin, bone, cartilage, and tendon Group 6: The group consisting of fish bones, skin, and scales.

[0015] 〔Biological function regulator〕 The biological function regulator according to this embodiment contains fermented collagen peptide. The above fermented collagen peptide has at least one action selected from the group consisting of an action of promoting epidermal metabolism, an action of suppressing fat accumulation, an action of promoting fat decomposition, and an action of regulating the amount of adipocytokines in the living body. The biological function regulator having such characteristics can exert at least one action (hereinafter, also referred to as "biological function regulating action") selected from the group consisting of an action of promoting epidermal metabolism, an action of suppressing fat accumulation, an action of promoting fat decomposition, and an action of regulating the amount of adipocytokines in the living body on the living body.

[0016] <Fermented collagen peptide> The biological function regulator according to this embodiment contains fermented collagen peptide as described above. The fermented collagen peptide preferably contains a collagen peptide and at least one first compound selected from the group consisting of isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional. The fermented collagen peptide more preferably contains a collagen peptide and at least three first compounds selected from the group consisting of isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional. Thereby, at least one biological function regulating action selected from the group consisting of an epidermal metabolism promoting action, a fat accumulation suppressing action, a fat decomposition promoting action, and an action of regulating the amount of adipocytokines in the living body can be exhibited more sufficiently. Further, as will be described later, the biological function regulator according to this embodiment is produced by fermenting a collagen raw material with koji. That is, the biological function regulator according to this embodiment is a biological function regulator containing a fermented collagen peptide.

[0017] Here, the odor (so-called collagen odor) of the conventional collagen peptide is suppressed by the first compound in the fermented collagen peptide. Therefore, the biological function regulator can be used, for example, in an epidermal metabolism promoter, a fat accumulation inhibitor, a fat decomposition promoter, an adiponectin production promoter, a functional food, and a cosmetic described later, by simplifying the deodorization treatment or the like, or without performing the deodorization treatment or the like.

[0018] (Collagen peptide) The fermented collagen peptide preferably contains the collagen peptide as described above. This collagen peptide appears to be the same as the conventionally known collagen peptide. That is, the collagen peptide contained in the fermented collagen peptide may contain various peptides such as dipeptides, tripeptides, oligopeptides, and polypeptides obtained by performing conventionally known treatments on collagen or gelatin as a peptide mixture apparently. However, the fermented collagen peptide is obtained by fermenting a collagen raw material with koji. Therefore, the collagen peptide contained in the above fermented collagen peptide is obtained by fermenting a collagen raw material with koji and is produced together with the first compound described later.

[0019] 〈Weight-average molecular weight〉 The collagen peptide contained in the fermented collagen peptide preferably has a weight-average molecular weight of 20,000 or less. When the weight-average molecular weight of the above collagen peptide is 20,000 or less, the biological function regulator can be easily applied to each use of an epidermal metabolism promoter, a fat accumulation inhibitor, a fat decomposition promoter, an adiponectin production promoter, a functional food, and a cosmetic without performing additional treatment. The above collagen peptide more preferably has a weight-average molecular weight of 10,000 or less, and even more preferably 6,000 or less. The lower limit value of the weight-average molecular weight of the above collagen peptide is 76. When the weight-average molecular weight of the above collagen peptide is within the above range, the biological function regulator can sufficiently exert the above-described effects on the living body in each use of an epidermal metabolism promoter, a fat accumulation inhibitor, a fat decomposition promoter, an adiponectin production promoter, a functional food, or a cosmetic.

[0020] Here, the weight-average molecular weight of the above collagen peptide contained in the above biological function regulator can be determined by performing size exclusion chromatography (SEC) under the following measurement conditions. The inventors have confirmed that this measurement method is valid for the measurement of molecular weights exceeding 12,000. Apparatus: High Performance Liquid Chromatography (HPLC) (manufactured by Tosoh Corporation) Column: TSKGel (registered trademark) G2000SW XL Column temperature: 40 °C Eluent: 45% by mass acetonitrile (containing 0.1% by mass TFA) Flow rate: 1.0 mL / min Injection volume: 10 μL Detection: UV 214 nm Molecular weight markers: The following 5 types were used Cytochrom C Mw: 12000 Aprotinin Mw: 6500 Bacitracin Mw: 1450 Gly-Gly-Tyr-Arg Mw: 451 Gly-Gly-Gly Mw: 189

[0021] (First compound) The fermented collagen peptide preferably contains at least one first compound selected from the group consisting of isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional as described above, more preferably contains at least three of the above first compounds. In the above fermented collagen peptide, the first compound is considered to be produced together with the collagen peptide by fermenting the collagen raw material with koji. The first compound can function as a marker indicating that the above fermented collagen peptide is obtained by fermenting the collagen raw material with koji.

[0022] The fermented collagen peptide may contain, as the first compound, any one of isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional. The fermented collagen peptide may contain isovaleraldehyde and 1-octen-3-ol as the first compounds, may contain isovaleraldehyde and phenylacetaldehyde, may contain isovaleraldehyde and methional, may contain 1-octen-3-ol and phenylacetaldehyde, may contain 1-octen-3-ol and methional, or may contain phenylacetaldehyde and methional.

[0023] The fermented collagen peptide may contain, as the first compounds, isovaleraldehyde, 1-octen-3-ol, and phenylacetaldehyde; isovaleraldehyde, 1-octen-3-ol, and methional; isovaleraldehyde, phenylacetaldehyde, and methional; or 1-octen-3-ol, phenylacetaldehyde, and methional. The fermented collagen peptide may also contain the four types of first compounds (isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional). In these cases, the biological function regulator can more sufficiently exhibit at least one action selected from the group consisting of an action of promoting epidermal metabolism, an action of suppressing fat accumulation, an action of promoting fat decomposition, and an action of regulating the amount of adipocytokines in the living body on the living body.

[0024] (Isovaleraldehyde) Isovaleraldehyde is a compound also known as isovaleric aldehyde, 3-methylbutanal, or 3-methylbutyl aldehyde, and is a compound that has conventionally been used as a fragrance (food additive) and the like.

[0025] (1-Octen-3-ol) 1-Octen-3-ol is a kind of unsaturated alcohol and is a compound known to contribute to the aroma of matsutake mushrooms conventionally.

[0026] (Phenylacetaldehyde) Phenylacetaldehyde is a kind of aromatic aldehyde and is a compound that has been conventionally used as a raw material for blending fragrances and flavors, etc.

[0027] (Methional) Methional is a kind of organic sulfur compound and is a compound also called 3-methylthio-1-propanol. Methional is a compound known to be contained in soy sauce conventionally. Furthermore, methional is also known to have an effect of weakening the fishy smell of meat and fish. Na

[0028] (Content) The first compound is preferably contained in the above biological function regulator at 0.05 ppm or more as its total amount (the total of at least one or more). That is, the above biological function regulator preferably contains the above first compound at 0.05 ppm or more. Furthermore, the first compound is more preferably contained in the above biological function regulator at 0.4 ppm or more as its total amount. That is, the above biological function regulator more preferably contains the above first compound at 0.4 ppm or more. The lower limit of the content of the first compound in the above biological function regulator is not particularly limited, but it is preferably contained at 0.01 ppm or more as the total amount of the first compound. The upper limit of the content of the first compound is not particularly limited either, but it is preferably 5 ppm or less as the total amount of the first compound.

[0029] The qualitative and quantitative determination of the first compound contained in the above biological function regulator can be determined by the following procedure. First, a dry powder of the biological function regulator is obtained by the production method described below. Further, a measurement sample is obtained by dissolving 0.5 g of the above dry powder in 4.5 mL of RO water. Next, the above measurement sample is introduced into a gas chromatograph mass spectrometer (trade name: "7890A GC System", manufactured by Agilent Technologies, Inc., and trade name: "JMS-Q1050GC", manufactured by JEOL Ltd.). After vaporizing this, the components contained in the above measurement sample are separated for each compound by moving it to a column provided in the above analyzer using ultra-high purity helium as the carrier gas. Further, the above compound is detected by a detector provided in the above analyzer, and the qualitative determination of the first compound can be performed by comparing the data (spectrum data) obtained from the above detector with the standard data. At the same time, the quantification of the first compound can be performed based on the above spectrum data (peak area) obtained from the above detector.

[0030] 〔Epidermal metabolism promoter〕 The epidermal metabolism promoter according to this embodiment contains the above biological function regulator. The epidermal metabolism promoter can promote, for example, the excretion of melanin granules on the skin surface (in the epidermis) from the skin surface without stagnating as spots, freckles, etc. by the epidermal metabolism promoting action of the above biological function regulator. Specifically, the above epidermal metabolism promoter can enhance the expression level of at least one gene selected from the group consisting of transglutaminase 1 (TGM1), involucrin (Ivl), and keratin 10 (KRT10). These genes are known to contribute to the maturation and differentiation of each layer (stratum corneum, granular layer, spinous layer, and basal layer) that constitutes the epidermis, and thus it is considered that the metabolism (so-called turnover) in the epidermis is promoted by the enhanced expression of the above genes. Furthermore, based on the promotion of metabolism in the epidermis, there is a possibility of obtaining effects such as a skin moisturizing effect, a prevention of skin wrinkles, and / or an improvement of skin wrinkles.

[0031] The concentration of the biological function regulator in the above-mentioned epidermis metabolism promoter may be 0.01 to 100% by mass. Since the content of the first compound in the above-mentioned epidermis metabolism promoter is extremely small, the concentration of the collagen peptide in the above-mentioned biological function regulator is meant. Therefore, the concentration of the biological function regulator in the above-mentioned epidermis metabolism promoter can be determined by a conventionally known method for measuring the concentration of collagen peptide. For example, the concentration of the biological function regulator in the above-mentioned epidermis metabolism promoter can be determined by measuring the mass percentage of hydroxyproline in the collagen peptide by the chloramine T method. Furthermore, it is also possible to determine it by using an amino acid analyzer to measure the mass percentage of hydroxyproline in the collagen peptide.

[0032] Adipose accumulation inhibitor The adipose accumulation inhibitor according to this embodiment contains the above-mentioned biological function regulator. The adipose accumulation inhibitor can obtain an inhibitory effect on the accumulation of fat (so-called visceral fat) in the liver, intestinal tract, kidney, testis, etc. due to the adipose accumulation inhibitory action of the above-mentioned biological function regulator. Also, the adipose accumulation inhibitor is based on the action of regulating the amount of adipocytokines in the living body that the above-mentioned biological function regulator has, and Inhibition through the action of decreasing the blood concentration of leptin, which is known as a hormone, and the action of increasing the blood concentration of adiponectin, which is known to be inversely correlated with the amount of visceral fat in the body, an adipose accumulation inhibitory effect can be obtained. The above-mentioned leptin and adiponectin are known as proteins classified as adipocytokines secreted from adipocytes. That is, the adiponectin production promoter according to this embodiment contains the above-mentioned biological function regulator.

[0033] The concentration of the biological function regulator in the above-mentioned fat accumulation inhibitor may be 0.01 to 100% by mass. The concentration of the biological function regulator in the above-mentioned adiponectin production promoter can also be 0.01 to 100% by mass. Since the content of the first compound in the above-mentioned fat accumulation inhibitor and adiponectin production promoter is very small, it shall mean the concentration of collagen peptide in the above-mentioned biological function regulator. Therefore, the concentration of the biological function regulator in the above-mentioned fat accumulation inhibitor and adiponectin production promoter can be the same as the method for measuring the concentration of the biological function regulator in the above-mentioned epidermis metabolism promoter described above.

[0034] The lipolysis promoter according to this embodiment contains the above-mentioned biological function regulator. The above-mentioned lipolysis promoter can obtain the effect of promoting the decomposition of fat (so-called visceral fat) in the liver, intestinal tract, kidney, testis, etc. by the lipolysis promoting action of the above-mentioned biological function regulator. The concentration of the biological function regulator in the above-mentioned lipolysis promoter may be 0.01 to 100% by mass. Since the content of the first compound in the above-mentioned lipolysis promoter is very small, it shall mean the concentration of collagen peptide in the above-mentioned biological function regulator. Therefore, the concentration of the biological function regulator in the above-mentioned lipolysis promoter can be the same as the method for measuring the concentration of the biological function regulator in the above-mentioned epidermis metabolism promoter described above.

[0035] Here, the above-mentioned epidermis metabolism promoter, fat accumulation inhibitor, lipolysis promoter and adiponectin production promoter can be administered orally or parenterally in various forms as supplements, pharmaceuticals or quasi-drugs. As its form, when administered orally, for example, dosage forms such as tablets, granules, capsules, powders, liquids, suspension preparations, emulsion preparations, pastes, etc. can be used. Furthermore, the above-mentioned dosage forms can also be mixed with the functional foods described later.

[0036] ​​When the above epidermal metabolism promoter, fat accumulation inhibitor, fat decomposition promoter, and adiponectin production promoter are administered parenterally, they can be formulated into, for example, injectables into the body, injections, transdermal agents (coating agents, patches, and aerosol agents), suppositories, nasal drops, and inhalants. Preferred dosage forms of the above epidermal metabolism promoter, fat accumulation inhibitor, fat decomposition promoter, and adiponectin production promoter include tablets, granules, capsules, powders, liquids, and transdermal agents, etc.

[0037] The dosage of the above epidermal metabolism promoter, fat accumulation inhibitor, fat decomposition promoter, and adiponectin production promoter varies depending on the age, sex, weight, sensitivity difference, administration method, administration interval, type of formulation, etc. of the subject. When the above epidermal metabolism promoter, fat accumulation inhibitor, fat decomposition promoter, and adiponectin production promoter are administered orally, the dosage for each is preferably, for example, 0.0001 to 2500 mg / kg per day for adults, and more preferably 0.0001 to 500 mg / kg per day. When the dosage form of the above epidermal metabolism promoter, fat accumulation inhibitor, fat decomposition promoter, and adiponectin production promoter is, for example, a tablet, it can be a tablet containing 0.001 to 80% by mass of the epidermal metabolism promoter, fat accumulation inhibitor, fat decomposition promoter, or adiponectin production promoter per tablet. When it is, for example, a powder, it can be a powder containing 0.001 to 100% by mass of the epidermal metabolism promoter, fat accumulation inhibitor, fat decomposition promoter, or adiponectin production promoter. When the above epidermal metabolism promoter, fat accumulation inhibitor, fat decomposition promoter, and adiponectin production promoter are administered parenterally, the dosage can be appropriately determined with reference to the dosage for oral administration. The above epidermal metabolism promoter, fat accumulation inhibitor, fat decomposition promoter, and adiponectin production promoter can be administered in 1 to several divided doses per day, or can also be administered once every 1 to several days.

[0038] The above-mentioned epidermis metabolism promoter, fat accumulation inhibitor, fat decomposition promoter, and adiponectin production promoter can appropriately contain other active ingredients, carriers for pharmaceutical preparations, etc., as long as they do not adversely affect the effects of the present invention. Examples of other active ingredients include (±)-α-tocopherol disodium phosphate salt, heparin-like substances, allantoin, glycyrrhizic acid, glycyrrhetin, D-form amino acids, aminosilane compounds, tiliroside, yuzu extract, α-glucosyl hesperidin, mulberry leaf extract, mangosteen pericarp extract, α-mangostin, γ-mangostin, cocoa seed extract, cocoa husk extract, Korean ginseng extract, lychee polyphenol, extract of Psophocarpus, extract of Gynura procumbens (Lour.) Merr., lactic acid bacteria, grabrutin, arbutin extract, extract of dried kudzu flower, extract of Juncus effusus Extract of Tricholoma matsutake , crocetin extract, N-acetylglucosamine, anserine, raspberry ketone, etc. can be mentioned. Furthermore, examples of pharmaceutically acceptable carriers used when formulating into pharmaceutical preparations include diluents, binders (syrup, gum arabic, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone), excipients (lactose, sucrose, corn starch, potassium phosphate, sorbitol, glycine), lubricants (magnesium stearate, talc, polyethylene glycol, silica), disintegrants (potato starch) and wetting agents (sodium lauryl sulfate), etc.

[0039] [Functional Food] The functional food according to this embodiment contains the above-mentioned biological function regulator. Examples of the above-mentioned functional food include foods for specified health use and foods with functional claims. The above-mentioned functional food can exhibit at least one action selected from the group consisting of an epidermal metabolism promoting action, a fat accumulation suppressing action, a fat decomposition promoting action, and an action of regulating the amount of adipocytokine in the living body, which the above-mentioned biological function regulator has, for example, as a food for specified health use or a food with functional claims. The concentration of the biological function regulator in the above-mentioned functional food can be 0.01 to 100% by mass, for example, in the above-mentioned food for specified health use or food with functional claims. Since the content of the first compound is extremely small, the concentration of the biological function regulator in the above-mentioned functional food means the concentration of collagen peptide in the above-mentioned biological function regulator. Therefore, the concentration of the biological function regulator in the above-mentioned functional food can be the same as the method for measuring the concentration of the biological function regulator in the above-mentioned epidermal metabolism promoter described above.

[0040] 〔Cosmetics〕 The cosmetic according to this embodiment contains the above-mentioned biological function regulator. The above-mentioned cosmetic can be provided as a cosmetic having a whitening effect, a moisturizing effect, a wrinkle prevention and / or wrinkle improvement effect, etc., based on the epidermal metabolism promoting action that the above-mentioned biological function regulator has, for example. The concentration of the biological function regulator in the above-mentioned cosmetic may be 0.01 to 100% by mass. Since the content of the first compound is extremely small, the concentration of the biological function regulator in the above-mentioned cosmetic means the concentration of collagen peptide in the above-mentioned biological function regulator. Therefore, the concentration of the biological function regulator in the above-mentioned cosmetic can be the same as the method for measuring the concentration of the biological function regulator in the above-mentioned epidermal metabolism promoter described above.

[0041] 〔Method for producing biological function regulator〕 The method for manufacturing a biological function regulator according to this embodiment is a method for manufacturing a biological function regulator containing fermented collagen peptide. The method for manufacturing the biological function regulator includes a step of preparing koji containing Aspergillus and a collagen raw material (first step), and a step of obtaining a biological function regulator containing the fermented collagen peptide by fermenting the collagen raw material with the koji (second step).

[0042] In the method for manufacturing the biological function regulator, the strain of the Aspergillus is a strain belonging to the genus Aspergillus. The collagen raw material is at least one selected from the group consisting of the following first group to sixth group, collagen extracted from at least one selected from the group, gelatin obtained by treating the collagen, and at least any one of gelatin degradation products obtained by hydrolyzing the gelatin. First group: The group consisting of cowhide, skin, bone, cartilage and tendon Second group: The group consisting of pigskin, skin, bone, cartilage and tendon Third group: The group consisting of sheepskin, skin, bone, cartilage and tendon Fourth group: The group consisting of chicken skin, skin, bone, cartilage and tendon Fifth group: The group consisting of ostrich skin, skin, bone, cartilage and tendon Sixth group: The group consisting of fish bone, skin and scale.

[0043] The method for manufacturing a biological function regulator having such characteristics can manufacture a biological function regulator containing a fermented collagen peptide having at least one action selected from the group consisting of an epidermal metabolism promoting action, a fat accumulation suppressing action, a fat decomposition promoting action, and an adipocytokine amount adjusting action in vivo.

[0044] In the biological function regulator produced by the above manufacturing method, the reason why it can have at least one action selected from the group consisting of an epidermal metabolism promoting action, a fat accumulation inhibitory action, a fat decomposition promoting action, and an action of regulating the amount of adipocytokines in the living body is not clear in detail, but it is considered to be due to the following mechanism. That is, the above manufacturing method includes a step (second step) of obtaining a biological function regulator containing fermented collagen peptides by fermenting a collagen raw material with koji. The above koji is known to contain a variety of enzymes produced by the propagation of koji mold. Therefore, in the second step, these various enzymes may act to decompose or oxidize-reduce polypeptides in the collagen raw material and carbohydrates in the koji.

[0045] As a result, in the second step, when fermented collagen peptides are produced by the action of the above-described various enzymes, it is presumed that the fermented collagen peptides contain collagen peptides including dipeptides, tripeptides, oligopeptides, or polypeptides having at least one of the above-described epidermal metabolism promoting action, fat accumulation inhibitory action, fat decomposition promoting action, and action of regulating the amount of adipocytokines in the living body. It is also presumed that when fermented collagen peptides are produced by the action of the above-described various enzymes, the fermented collagen peptides contain a compound (non-peptide) having at least one of the above-described physiological activities. Thus, it is considered that a biological function regulator containing fermented collagen peptides having at least one action selected from the group consisting of an epidermal metabolism promoting action, a fat accumulation inhibitory action, a fat decomposition promoting action, and an action of regulating the amount of adipocytokines in the living body can be obtained by the above manufacturing method. Hereinafter, each step in the manufacturing method of the biological function regulator according to the present embodiment will be described.

[0046] <First Step> The first step is a step of preparing koji containing Aspergillus oryzae and a collagen raw material. The first step is carried out for the purpose of preparing each material (koji containing Aspergillus oryzae and collagen raw material) necessary for manufacturing the above-mentioned biological function regulator.

[0047] (Collagen raw material) The collagen raw material may be at least any one of "itself" selected from the group consisting of the following first to sixth groups as described above, "collagen" extracted from at least one selected from the group consisting of the following first to sixth groups, "gelatin" obtained by treating the above collagen using a known method such as hot water extraction, and "gelatin hydrolyzate" obtained by hydrolyzing the above gelatin. First group: The group consisting of cowhide, skin, bone, cartilage and tendon Second group: The group consisting of pigskin, skin, bone, cartilage and tendon Third group: The group consisting of sheepskin, skin, bone, cartilage and tendon Fourth group: The group consisting of chicken skin, skin, bone, cartilage and tendon Fifth group: The group consisting of ostrich skin, skin, bone, cartilage and tendon Sixth group: The group consisting of fish bones, skin and scales.

[0048] That is, in the first step, it is preferable to prepare at least one selected from the group consisting of the above first to sixth groups, at least one selected from the group consisting of the above collagen, the above gelatin and the above gelatin hydrolyzate as the above collagen raw material. In the first step, one kind of collagen raw material selected from these may be prepared, or two or more kinds of collagen raw materials may be prepared in combination. The above groups consisting of the first to sixth groups, the above collagen, the above gelatin and the above gelatin hydrolyzate can all be prepared by conventionally known methods.

[0049] Here, it is more preferable to obtain the gelatin by performing pretreatment by acid treatment or alkali treatment, hot water extraction, purification treatment, and sterilization treatment in this order on collagen extracted from at least one selected from the group consisting of the above-mentioned first to sixth groups. Thereby, gelatin with high safety for the human body and the like can be prepared, and thus the biological function regulator to be produced in this embodiment can be applied to each of the above-mentioned epidermal metabolism promoters, fat accumulation inhibitors, fat decomposition promoters, adiponectin production promoters, functional foods, and cosmetics. Furthermore, such gelatin is also excellent in economy. The above-mentioned pretreatment by acid treatment or alkali treatment, hot water extraction, purification treatment, and sterilization treatment can all be carried out by conventionally known methods.

[0050] The above-mentioned gelatin hydrolyzate can be obtained by performing any one of hydrolysis using a conventionally known acid, hydrolysis using a base, hydrolysis using an enzyme, and hydrolysis using heating on the above-mentioned gelatin. The weight average molecular weight of the gelatin hydrolyzate is not particularly limited, but is preferably, for example, 20,000 or less, and more preferably 10,000 or less. The lower limit value of the weight average molecular weight of the above-mentioned gelatin hydrolyzate is 76. The weight average molecular weight of the above-mentioned gelatin hydrolyzate can be determined by the same measurement method as the weight average molecular weight of the above-mentioned collagen peptide.

[0051] (Koji containing Aspergillus oryzae) The koji containing Aspergillus can be prepared by a conventionally known method as long as the koji that can obtain the effects of the present embodiment is selected by performing the second step described later. That is, it can be obtained by inoculating Aspergillus, which is the mother koji, onto coarse grains such as rice, barley, wheat, or soybeans, and then propagating it in the above-mentioned rice, barley, wheat, or coarse grains. It is preferable to inoculate the above Aspergillus in an amount of 0.01 to 1% by mass based on the above rice, barley, wheat, or coarse grains. In this specification, "coarse grains" includes all of bran, rice bran, okara, and defatted soybeans in addition to the above-mentioned soybeans. In the preparation of koji containing Aspergillus, it is preferable to prepare an environment in which Aspergillus easily propagates by providing a koji room for preventing the contamination of other bacteria, and perform necessary operations in the above koji room.

[0052] The bacterial species of the above Aspergillus is preferably a bacterial species belonging to the genus Aspergillus. The bacterial species of the above Aspergillus is more preferably at least one selected from the group consisting of Aspergillus sojae, Aspergillus oryzae, and Aspergillus luchuensis. Since these bacterial species have been confirmed to be safe for the human body, etc., the biological function regulator produced in the present embodiment can be easily applied to each of the above-mentioned epidermal metabolism promoters, fat accumulation inhibitors, fat decomposition promoters, adiponectin production promoters, functional foods, and cosmetics. In the first step, koji containing one selected from the group of these bacterial species may be prepared, or koji containing two or more selected from the group of the above bacterial species may be prepared.

[0053] <Second Step> The second step is a step of obtaining a biological function regulator containing the fermented collagen peptide by fermenting the collagen raw material with the koji. The second step is carried out for the purpose of obtaining the fermented collagen peptide contained in the biological function regulator. In the second step, for example, the collagen raw material and the koji are put into warm water, and these are cultured in warm water for a predetermined time to ferment the collagen raw material with the koji, whereby a biological function regulator containing the fermented collagen peptide can be obtained. The pH value during the culture is preferably from 2 to 10, more preferably from 5 to 8. If the pH value during the culture is less than 2 or exceeds 10, there is a risk that the reduction of the molecular weight of the collagen raw material, the reduction of the collagen odor, etc. may be insufficient.

[0054] Specifically, a dispersion liquid is prepared from 0.1 to 75% by mass of the collagen raw material, 0.1 to 20% by mass of the koji as the dry mass (dry weight), and 5 to 99.8% by mass of water, with a total of 100% by mass. After further adjusting the dispersion liquid to have a pH of 2 to 10, it is preferable to culture the dispersion liquid for 1 to 24 hours while maintaining the temperature of the dispersion liquid at 10 to 65 °C. Thus, a fermented product containing the fermented collagen peptide can first be obtained.

[0055] The above fermented product is also preferably obtained by the following method. That is, first, from the above koji with a dry mass (dry weight) of 0.1 to 40% by mass and 60 to 99.9% by mass of water, a dispersion in which their total is 100% by mass is prepared, and while maintaining the temperature of the above dispersion at 10 to 65°C, it is cultured for 1 to 24 hours, followed by rough filtration through a nylon mesh and filtration with diatomaceous earth and cellulose to obtain a koji extract. Next, from 0.1 to 75% by mass of the above collagen raw material and 0.1 to 99.9% by mass of the above koji extract, a dispersion in which their total is 100% by mass is prepared, and after further adjusting the above dispersion to have a pH of 2 to 10, it is cultured for 1 to 24 hours while maintaining the temperature of the above dispersion at 10 to 65°C. By this method as well, a fermented product containing fermented collagen peptides can be obtained. When a fermented product is obtained by applying this method, a biological function regulator containing fermented collagen peptides can be obtained without performing the separation treatment step described below on the fermented product. However, it does not exclude performing at least either the purification step or the deodorization step described below on the fermented product.

[0056] Here, the temperature of the warm water during culturing is preferably 15 to 60°C, more preferably 20 to 50°C. When the temperature of the warm water during culturing is below 10°C or exceeds 65°C, there is a possibility that the efficiency of fermentation by koji decreases, and fermented collagen peptides may not be sufficiently obtained.

[0057] Furthermore, the culturing time is preferably 2 to 18 hours, more preferably 4 to 8 hours. When the culturing time exceeds 24 hours, there is a risk of being economically inefficient. When the culturing time is less than 1 hour, there is a risk that the fermentation by koji is insufficient.

[0058] The content of the collagen raw material in the warm water during culturing is preferably 10 to 45% by mass, more preferably 20 to 40% by mass. When the content of the collagen raw material in the warm water during culturing is less than 0.1% by mass, there is a risk of being economically inefficient. When the content of the collagen raw material in the warm water during culturing exceeds 75% by mass, there is a risk that the operation becomes inefficient.

[0059] The content of koji in the dispersion liquid composed of the above koji, the above collagen raw material and water is preferably 1 to 15% by mass, more preferably 5 to 10% by mass, in terms of dry mass (dry weight). When the content of koji in warm water during cultivation is less than 0.1% by mass in terms of dry mass (dry weight), there is a risk that the fermentation by koji may be insufficient. When the content of koji in warm water during cultivation exceeds 20% by mass in terms of dry mass (dry weight), it may be economically inefficient. The content of koji in the above koji extract is preferably 2 to 25% by mass, more preferably 8 to 16% by mass. When the content of koji in the koji extract is less than 0.1% by mass in terms of dry mass (dry weight), there is a risk that the fermentation by koji may be insufficient. When the content of koji in the koji extract exceeds 40% by mass in terms of dry mass (dry weight), it may be economically inefficient.

[0060] In the second step, after obtaining the above fermented product by the above steps, the temperature is set to 75°C or higher according to the purpose, so that the action (activity) of koji mold can be inactivated, and thus the progress of the fermentation of the collagen raw material by koji can be stopped. Specifically, the weight average molecular weight of the collagen peptide in the above fermented product is measured, and it is confirmed that the molecular weight has been reduced compared to the collagen raw material, or when the cultivation time has elapsed for a predetermined time, for example, 24 hours, the temperature of the above fermented product is set to 75°C or higher to stop the progress of the fermentation of the collagen raw material by koji. The weight average molecular weight of the collagen peptide in the fermented product can be, for example, the same as the measurement method of the weight average molecular weight of the collagen peptide described above.

[0061] (Other steps) The second step preferably includes a separation treatment step for separating a biological function regulator containing fermented collagen peptide from the above-mentioned fermented product. This separation treatment step can apply conventionally known separation treatments. For example, through separation treatments such as rough filtration using a nylon mesh, centrifugation, and filter paper filtration using commercially available filter paper, the fermented collagen peptide can be separated from the above-mentioned fermented product. Thereby, preferably, a biological function regulator containing a fermented collagen peptide containing collagen peptide and at least one, more preferably three, first compounds selected from the group consisting of isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional can be obtained. Since the above-mentioned fermented product contains fermented collagen peptide, the fermented product itself can also be regarded as a biological function regulator.

[0062] Furthermore, the second step preferably also includes a purification step (purification process) of purifying the biological function regulator obtained by applying the above-mentioned separation treatment step or the above-mentioned fermented product for the purpose of increasing its transparency. In this purification step, conventionally known purification treatments can be applied, for example, purification treatment using diatomaceous earth or purification treatment by microfiltration can be performed. Furthermore, deodorization treatment (deodorization process) can also be performed by using activated carbon or the like.

[0063] The biological function regulator obtained as described above can be stored as it is in a solution state. Furthermore, for the biological function regulator in a solution state, a dry powder of the biological function regulator can be obtained by using conventionally known methods such as spraying or drum drying, and it can also be stored in that state. Furthermore, by using conventionally known pharmaceutical technologies for the dry powder of the above-mentioned biological function regulator, it is possible to obtain various dosage forms as described above.

[0064] <Function and Effect> From the above, the method for producing a biological function regulator according to the present embodiment includes fermented collagen peptide, and based on the above fermented collagen peptide, at least one biological function regulating action selected from the group consisting of an epidermal metabolism promoting action, a fat accumulation suppressing action, a fat decomposition promoting action, and an adipocytokine amount regulating action in vivo can be obtained.

Example

[0065] Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto. In the following description, Samples 1 to 5 and Samples 41 to 49 are biological function regulators of the examples, and Samples 101 to 104 are collagen peptides or gelatin of the comparative examples.

[0066] 〔Preparation of Samples〕 <Sample 1> (First Step) According to the following procedure, koji containing Aspergillus and a collagen raw material were prepared.

[0067] 〈Preparation of Koji Containing Aspergillus〉 As koji containing Aspergillus, barley bran koji inoculated with Aspergillus sojae (manufactured by Higuchi Matsunosuke Shoten Co., Ltd.) was prepared.

[0068] 〈Preparation of Collagen Raw Material〉 As a collagen raw material, gelatin derived from pig skin (trade name: "BCN-HL", manufactured by Nitta Gelatin Inc., weight average molecular weight: about 65,000) was prepared.

[0069] (Second Step) The fermented product containing fermented collagen peptides was obtained by fermenting the above collagen raw material with the above koji. First, a dispersion consisting of 5 g of the above collagen raw material, 1 g (dry weight) of the above barley bran koji, and 50 mL of RO water was prepared, and cultured for 5 hours while maintaining the temperature of the dispersion at 40°C. Then, the temperature of the dispersion was set to 75°C, and the koji mold in the above barley bran koji was inactivated by maintaining the dispersion at a temperature near 75°C for 10 minutes, thereby obtaining a fermented product containing fermented collagen peptides.

[0070] Next, the above fermented product was filtered using ADVANTEC FILTER PAPER No. 2 (manufactured by Toyo Roshi Kaisha, Ltd.) to obtain a biological function regulator for Sample 1.

[0071] The biological function regulator for Sample 1 is an aqueous solution. When its weight-average molecular weight was measured, it was confirmed that the molecular weight was reduced compared to the weight-average molecular weight of the above collagen raw material. Also, from the analysis using the above gas chromatograph-mass spectrometer, it was confirmed that the biological function regulator for Sample 1 contains isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional as the first compounds.

[0072] <Sample 2> In the second step, a dispersion consisting of 1 kg of the above collagen raw material, 200 g (dry weight) of the above barley bran koji, and 1500 mL of RO water was prepared, and cultured for 6 hours while maintaining the temperature of the dispersion at 40°C. Then, the temperature of the dispersion was set to 70°C, and the koji mold in the above barley bran koji was inactivated by maintaining the dispersion at a temperature near 70°C for 1 hour, thereby obtaining a fermented product containing fermented collagen peptides. Further, the above fermented product was filtered using ADVANTEC FILTER PAPER No. 5 (manufactured by Toyo Roshi Kaisha, Ltd.) and further purified by diatomaceous earth filtration. Otherwise, the biological function regulator for Sample 2 was obtained by the same method as for obtaining Sample 1.

[0073] Regarding the biological function regulator of Sample 2, it was made into a dry powder by using a spray dryer (manufactured by Okawara Seisakusho Co., Ltd.).

[0074] The biological function regulator of Sample 2 is a dry powder. When its weight-average molecular weight was measured, it was confirmed that it had a lower molecular weight compared to the weight-average molecular weight of the above collagen raw material. Also, from the analysis using the above-mentioned gas chromatograph-mass spectrometer, it was confirmed that the biological function regulator of Sample 2 contains isovaleraldehyde, phenylacetaldehyde, and methional as the first compound.

[0075] <Sample 3> (First step) According to the following procedure, koji containing Aspergillus oryzae and a collagen raw material were prepared.

[0076] 〈Preparation of koji containing Aspergillus oryzae〉 As koji containing Aspergillus oryzae, barley bran koji inoculated with Aspergillus sojae (manufactured by Higuchi Matsunosuke Shoten Co., Ltd.) was prepared.

[0077] 〈Preparation of collagen raw material〉 As the collagen raw material, gelatin derived from pig skin (trade name: "BCN-HL", manufactured by Nitta Gelatin Inc., weight-average molecular weight: about 65,000) was prepared.

[0078] (Second step) By following the steps below, the above collagen raw material was fermented with the above koji to obtain a fermented product containing fermented collagen peptides. First, a dispersion consisting of 13% by mass (dry weight) of the above barley bran koji and 87% by mass of RO water was prepared, and the dispersion was stirred for 1 hour while maintaining the temperature of the dispersion at 40°C. Then, a koji extract was obtained by performing rough filtration through a nylon mesh and filtration with diatomaceous earth and cellulose. Next, a dispersion consisting of 40% by mass of the above collagen raw material and 60% by mass of the above koji extract was prepared, and the dispersion was cultured for 6 hours while maintaining the temperature of the dispersion at 40°C. Then, the temperature of the dispersion was set to 80°C, and pasteurization was performed by maintaining the dispersion at a temperature near 80°C for 60 minutes. Moreover, a dry powder was obtained by using a spray dryer (manufactured by Okawara Seisakusho Co., Ltd.), and thus a biological function regulator of Sample 3 was obtained.

[0079] The biological function regulator of Sample 3 is a dry powder. When its weight-average molecular weight was measured, it was confirmed that the molecular weight was reduced compared to the weight-average molecular weight of the above collagen raw material. Also, from the analysis using the above gas chromatograph mass spectrometer, it was confirmed that the biological function regulator of Sample 3 contains isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional as the first compounds.

[0080] <Sample 4> A dispersion consisting of 40% by mass of the above collagen raw material and 60% by mass of the above koji extract was prepared, and after culturing the dispersion for 6 hours while maintaining the temperature of the dispersion at 40°C, the temperature of the dispersion was set to 60°C, and pasteurization was performed by maintaining the dispersion at a temperature near 60°C for 60 minutes. A biological function regulator of Sample 4 was obtained by the same method as obtaining Sample 3, except for the above.

[0081] The biological function regulator of Sample 4 is a dry powder. When its weight-average molecular weight was measured, it was confirmed that the molecular weight was reduced compared to the weight-average molecular weight of the above collagen raw material. Also, from the analysis using the above gas chromatograph mass spectrometer, it was confirmed that the biological function regulator of Sample 4 contains isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional as the first compounds.

[0082] <Sample 5> (First step) Aspergillus oryzae-containing koji and a collagen raw material were prepared according to the following procedure.

[0083] 〈Preparation of Aspergillus oryzae-containing koji〉 As the Aspergillus oryzae-containing koji, barley bran koji inoculated with Aspergillus sojae (manufactured by Higuchi Matsunosuke Shoten Co., Ltd.) was prepared.

[0084] 〈Preparation of collagen raw material〉 As the collagen raw material, gelatin derived from tilapia scales (manufactured by Nitta Gelatin Inc., weight average molecular weight: about 150,000) was prepared.

[0085] (Second step) According to the following procedure, the above collagen raw material was fermented with the above koji to obtain a fermented product containing fermented collagen peptide. First, a dispersion consisting of 10% by mass of the above collagen raw material, 2% by mass (dry weight) of the above barley bran koji, and 88% by mass of RO water was prepared, and the dispersion was cultured for 6 hours while maintaining the temperature of the dispersion at 40°C. Then, the temperature of the dispersion was set to 75°C, and the dispersion was maintained at a temperature near 75°C for 60 minutes to inactivate the koji mold in the above barley bran koji, thereby obtaining a fermented product containing fermented collagen peptide.

[0086] Next, the above fermented product was centrifuged at a centrifugal acceleration of 1610 G for 30 minutes, and the supernatant was obtained to obtain a biological function regulator of Sample 5.

[0087] The biological function regulator of Sample 5 is an aqueous solution. When its weight average molecular weight was measured, it was confirmed that the weight average molecular weight was lower than that of the above collagen raw material. In addition, from the analysis using the above-mentioned gas chromatograph-mass spectrometer, it was confirmed that the biological function regulator of Sample 5 contains isovaleraldehyde, 1-octen-3-ol, and phenylacetaldehyde as the first compound.

[0088] <Sample 101> The dry powder of collagen peptide (trade name: "Collapep PU", manufactured by Nitta Gelatin Inc., weight-average molecular weight: 630) was prepared as Sample 101. Sample 101 was confirmed to contain no first compound from the analysis using the above-described gas chromatograph mass spectrometer.

[0089] <Sample 102> The dry powder of gelatin derived from porcine skin (trade name: "BCN-HL", manufactured by Nitta Gelatin Inc., weight-average molecular weight: approximately 65,000) was prepared as Sample 102. Sample 102 does not contain collagen peptide and was confirmed to contain no first compound from the analysis using the above-described gas chromatograph mass spectrometer.

[0090] <Sample 103> The dry powder of collagen peptide (trade name: "CP Prototype", manufactured by Nitta Gelatin Inc., weight-average molecular weight: 500 - 1000) was prepared as Sample 103. Sample 103 was confirmed to contain no first compound from the analysis using the above-described gas chromatograph mass spectrometer.

[0091] <Sample 104> The dry powder of collagen peptide (trade name: "SCP-5200", manufactured by Nitta Gelatin Inc., weight-average molecular weight: 3000 - 6000) was prepared as Sample 104. Sample 104 was confirmed to contain no first compound from the analysis using the above-described gas chromatograph mass spectrometer.

[0092] [First Test] By administering the biological function regulator of Sample 2 to mice, it was tested whether the above biological function regulator has an effect of suppressing fat accumulation. Specifically, the first test was carried out according to the following procedure.

[0093] <Test Method> Thirty 5-week-old male C57BL / 6J mice were prepared by purchasing them from CLEA Japan, Inc. The above mice were divided into three groups (n = 10) of a low-fat diet intake group (hereinafter also referred to as the "L group"), a high-fat diet intake group (hereinafter also referred to as the "H group"), and a high-fat diet and sample 2 (5% by mass) intake group (hereinafter also referred to as the "FCP group"), and breeding (pair-feeding breeding) was carried out for 30 days by feeding each group with the corresponding feed. During the breeding, the feed intake and body weight of the mice were measured daily at a predetermined time. The composition of the feed (unit: mass%) given to the mice in each of the above groups is shown in Table 1. In addition, in Table 1, the composition of the feed used in the second test described later is also specified.

[0094]

Table 1

[0095] Next, after anesthetizing the above mice in each group with isoflurane, decapitation blood collection and dissection were performed Internal organs to obtain fat, serum, and liver. As the above visceral fat, mesenteric fat, perirenal fat, and peritesticular fat were excised. After measuring the fat amount (mass (g)) in each organ, the total Internal organs fat amount was determined by summing them up. Furthermore, the concentrations of leptin and adiponectin in the serum were quantified from the above serum. As the quantification method, the ELISA method was used, and the leptin concentration was determined according to the protocol of a mouse leptin measurement kit (catalog number: "MS333", manufactured by Morinaga Institute of Biological Science, Inc.). The adiponectin concentration was determined according to the protocol of Revis high molecular adiponectin-mouse / rat (catalog number: "634-13071", manufactured by Fujifilm Wako Pure Chemical Corporation).

[0096] For the liver, the following procedures were performed. First, 0.5 g of the above liver was homogenized with 2.5 mL of 1.15% by mass potassium chloride (KCl) solution and placed in a test tube cooled with ice. Further, the above test tube was centrifuged (gravitational acceleration 9830 G, 10 minutes, 4 °C), and the supernatant was used as a crude enzyme solution. Next, based on the methods described by Nepokroeff et al. (Methods Enzymol, 35: 37-44, 1975) and Kelley et al. (Biochem. J, 235: 87-90, 1986), the enzyme activity was measured (wavelength 340 nm) from the rate of decrease in NAPDH in the presence of 100 μM malonyl CoA and 25 μM acetyl CoA with respect to the above crude enzyme solution, and the strength of fatty acid synthase (FAS) activity was determined. Further, based on the method described by Markwell et al. (J Biol. Chem, 248: 3426-3432, 1973), the enzyme activity was measured (wavelength 412 nm) from the reaction rate of dithiobisnitrobenzoic acid (DTNB) in the presence of 2 mM palmitoyl CoA and 125 mM L-carnitine with respect to the above crude enzyme solution, and the strength of fatty acid degrading enzyme (carnitine palmitoyl transferase: CPT) activity was determined. The results are shown in FIGS. 1 to 9. Regarding the significant differences among the groups (L group, H group, and FCP group) in FIGS. 1 to 9, a Bonferrioni multiple comparison test was performed, and P < 0.05 was determined to be statistically significant.

[0097] <Discussion> FIG. 1 is a graph showing the change in body weight of mice in each group in the first test. According to FIG. 1, it is understood that the increase in body weight of the FCP group is significantly suppressed compared to the H group.

[0098] FIG. 2 is a graph showing the amount of intestinal membrane fat of mice in each group in the first test. FIG. 3 is a graph showing the amount of perirenal fat of mice in each group in the first test. FIG. 4 is a graph showing the amount of peritesticular fat of mice in each group in the first test. FIG. 5 is a graph showing the total amount of fat of mice in each group in the first test. Internal organsIt is a graph showing the amount of fat. According to FIGS. 2 to 5, it is evaluated that the fat accumulation in each organ of the FCP group is significantly suppressed compared to that of the H group. Thus, it is suggested that the FCP group can obtain an effect of suppressing fat accumulation.

[0099] FIG. 6 is a graph showing the leptin concentration in the serum of the mice in each group in the first test. FIG. 7 is a graph showing the adiponectin concentration in the serum of the mice in each group in the first test. According to FIGS. 6 to 7, it is evaluated that the blood concentration of leptin in the FCP group is significantly decreased compared to that of the H group, and the blood concentration of adiponectin in the FCP group is significantly increased compared to that of the H group. Thus, it is suggested that the FCP group can obtain an effect of suppressing fat accumulation through the action of regulating the amount of adipocytokines in the living body.

[0100] FIG. 8 is a graph showing the strength of FAS activity in the liver of the mice in each group in the first test. FIG. 9 is a graph showing the strength of CPT activity in the liver of the mice in each group in the first test. According to FIGS. 8 to 9, it is evaluated that the synthesis of fatty acids in the FCP group is significantly suppressed compared to that of the H group, and the decomposition of fatty acids in the FCP group is significantly promoted compared to that of the H group. Thus, it is suggested that the FCP group can obtain an effect of suppressing fat accumulation.

[0101] [Second Test] By administering the biological function regulator of Sample 2 to mice, a test was conducted on whether the biological function regulator has an effect of suppressing fat accumulation. Specifically, the second test was carried out according to the following procedure.

[0102] [Test Method] Forty 5-week-old male C57BL / 6J mice were prepared by purchasing them from CLEA Japan, Inc. The above mice were divided into five groups (n = 8): group L, group H, group FCP, high-fat diet and sample 101 (5% by mass) intake group (hereinafter also referred to as "CP group"), and high-fat diet and sample 102 (5% by mass) intake group (hereinafter also referred to as "GL group"), and breeding (pair-feeding breeding) was carried out for 30 days by feeding each group with the corresponding feed. During the breeding, the feed intake and body weight of the mice were measured daily at a predetermined time. The composition of the feed given to the mice in each of the above groups was as shown in Table 1 above.

[0103] Next, the above mice in each group were anesthetized with isoflurane, and then decapitated for blood collection and dissection Internal organs to obtain fat and serum. Then, in the same manner as in the first test described above, Internal organs the fat content and total Internal organs fat content in each organ were determined from the above fat. Furthermore, in the same manner as in the first test described above, the concentrations of leptin and adiponectin in the above serum were quantified. The results of the fat content and total Internal organs fat content (unit: g) in each organ are shown in Table 2. The results of the leptin concentration and adiponectin concentration in the serum are shown in FIGS. 10 to 11. Regarding the significant differences between each group (group L, group H, group GL, group CP, and group FCP) in FIGS. 10 to 11, a Bonferroni multiple comparison test was performed, and P < 0.05 was determined to be statistically significant.

[0104]

Table 2

[0105] <Discussion> According to Table 2, it is evaluated that the fat accumulation in each organ of the FCP group is suppressed compared to that of the H group. Furthermore, the fat accumulation in each organ of the FCP group is also evaluated to be suppressed compared to that of the CP group and the GL group. Thus, it is suggested that the FCP group can obtain an effect of suppressing fat accumulation.

[0106] Figure 10 is a graph showing the leptin concentration in the serum of mice in each group in the second test. Figure 11 is a graph showing the adiponectin concentration in the serum of mice in each group in the second test. According to Figures 10 to 11, it is evaluated that the blood concentration of leptin in the FCP group is decreased compared to that in the H group, and the blood concentration of adiponectin in the FCP group is increased compared to that in the H group. Furthermore, it is evaluated that the blood concentration of leptin in the FCP group is decreased compared to that in the CP group and the GL group, and the blood concentration of adiponectin in the FCP group is increased compared to that in the CP group and the GL group. Thus, it is suggested that in the FCP group, a fat accumulation inhibitory effect can be obtained based on the regulatory action of the amount of adipocytokines in vivo.

[0107] [Third Test] The biological function regulators of Sample 1 and Sample 2, and Samples 101 and 103 were added to adipocytes differentiated from mouse-derived preadipocytes (3T3-L1, passage number: 5 Passages, 9 PDL) to test whether the biological function regulators of Sample 1 and Sample 2 have a fat accumulation inhibitory effect. Specifically, the third test was performed according to the following procedure.

[0108] [Test Method] The above mouse-derived preadipocytes (manufactured by the Research Resource Bank, resource number JCRB9014, Lot. No. 01282009) were precultured with a culture medium (DMEM / F12 medium (subculture medium, catalog number: "11330-032", manufactured by Gibco, containing 10% by mass FBS, penicillin and streptomycin)). Then, from the precultured medium and the above cells, a cell suspension was prepared such that the cells were 3×10 4 cells / mL. Further, 5 mL of the cell suspension was seeded into each dish of a 60 mm dish (catalog number: Corning, Cat. No. 430166) (1.5×10 5The cells were seeded in a 6-well plate at a density of 1×10⁵ cells / well (or in a 12-well plate at a density of 5×10⁴ cells / well or in a 24-well plate at a density of 2×10⁴ cells / well or in a 96-well plate at a density of 1×10⁴ cells / well or in a cell / dish), and cultured for 2 days under the conditions of 37°C (5% CO₂ by volume). Next, the medium in each dish was replaced with a differentiation induction medium supplemented with isobutylmethylxanthine (IBMX) and dexamethasone attached to an adipogenesis assay kit (catalog number: "ECM950", manufactured by Millipore) so that the final concentrations were 0.5 mM and 1 μM, respectively, and cultured for an additional 2 days under the conditions of 37°C (5% CO₂ by volume). Thereafter, the differentiation induction medium was further replaced with a differentiation medium supplemented with insulin attached to an adipogenesis assay kit (catalog number: "ECM950", manufactured by Millipore) so that the final concentration was 10 μg / mL, and cultured for 2 days at 37°C (5% CO₂ by volume). After confirming that the cells in each dish had differentiated into adipocytes, the differentiation medium was replaced with the subculture medium. To the adipocytes in this subculture medium, a biological function regulator of Sample 1 and Sample 2, and Sample 101 and Sample 103 were added so that the final concentration was 0.1% by mass, a berberine chloride solution (catalog number: "027-11781", manufactured by Fujifilm Wako Pure Chemical Corporation) was added so that the final concentration was 2 μg / mL, and RO water was added to the remaining adipocytes in the subculture medium. The cells were cultured for 2 days under the conditions of 37°C (5% CO₂ by volume). Note that Sample 1, Sample 2, Sample 101, Sample 103, the berberine chloride solution, and RO water were added to 3 dishes each. Thereafter, the subculture medium was replaced with a new subculture medium, Sample 1, Sample 2, Sample 101, and Sample 103 were re-added so that the final concentration was 0.1% by mass, the berberine chloride solution was re-added so that the final concentration was 2 μg / mL, and RO water was re-added to the corresponding dishes.

[0109] Next, according to the protocol of the adipogenesis assay kit, the Oil Red O solution attached to the kit was added to each dish at room temperature. That is, the adipocytes in each medium added with Sample 1, Sample 2, Sample 101, Sample 103, berberine chloride solution, and RO water were washed twice with PBS (phosphate buffered saline), and then 1.25 mL of the above Oil Red O solution with a concentration of 36% by mass was added to each dish. Then, it was washed twice with 2.5 mL of isopropanol with a concentration of 60% by mass, and 625 μL of isopropanol with a concentration of 99% by mass was added to obtain an extract containing adipocytes and Oil Red O from each dish. 200 μL of the above extract was transferred to a 96-well plate, and the absorbance at OD520nm was measured with an absorbance meter (product name: "Synergy HTX", manufactured by Biotech Japan Co., Ltd.). The higher the absorbance at OD520nm, the more it is evaluated that fat accumulation has occurred in adipocytes. Here, the extract obtained from the dish added with RO water becomes the Blank. Also, the extract obtained from the dish added with the berberine chloride solution becomes the positive control.

[0110] The fat accumulation rate in adipocytes added with each sample, berberine chloride solution, or RO water was determined based on the following calculation formula. Fat accumulation rate (%) = [(measured value of OD520nm of each sample) / (measured value of OD520nm of Blank) × 100].

[0111] The results are shown in Table 3. Table 3 shows the average value (Ave) and standard deviation of the fat accumulation rates of three specimens for each sample and the like. Furthermore, in Table 3, by performing statistical processing on each measured value, a determination was also made regarding the significant difference as to whether fat accumulation was suppressed. For the above statistical processing, analysis processing software (product name: "STAT Mate V", manufactured by Atomus Co., Ltd.) was used. Regarding the above significant difference, it was determined by performing one-way analysis of variance (One-way Anova) and Tukey test with a confidence interval of 95% confidence rate as post hoc tests. In Table 3, p < 0.001 is represented by ***, p < 0.01 is represented by **, and p < 0.05 is represented by *.

[0112]

Table 3

[0113] <Investigation> According to Table 3, the biological function regulators of Sample 1 and Sample 2 are evaluated to have suppressed fat accumulation in adipocytes compared to Sample 101 and Sample 103.

[0114] 〔Fourth Test〕 By adding the biological function regulators of Samples 41 to 49, which will be described later, to adipocytes differentiated from mouse-derived preadipocytes (3T3-L1, passage number: 7 Passages, 13 PDL), it was tested whether the biological function regulators of Samples 41 to 49 have an inhibitory effect on fat accumulation. Specifically, the Fourth Test was performed according to the following procedure.

[0115] <Test Method> (Preparation of Samples 41 to 49) The biological function regulators of Samples 41 to 49 were prepared by the same method as for obtaining the biological function regulator of Sample 2, except that the conditions for fermenting the collagen raw material with the above koji in the second step were as shown in Table 4. Note that for Sample 43, the conditions for fermenting the collagen raw material with the above koji were the same as those for Sample 2. Table 4 also shows the weight-average molecular weight of the collagen peptides contained in each sample.

[0116]

Table 4

[0117] (Test Method) The Fourth Test was performed in the same manner as the Third Test, except that the passage number of mouse-derived preadipocytes (3T3-L1) was set to "7 Passages, 13 PDL" and the final concentrations of Samples 41 to 49 added to the adipocytes in the above dish were set to 0.2% by mass. The results are shown in Table 5.

[0118]

Table 5

[0119] <Investigation> According to Table 5, the biological function regulators of Samples 41 to 49 are all evaluated to have suppressed fat accumulation in adipocytes compared to the Blank. In the fourth test, no determination was made regarding the presence or absence of a significant difference in the suppression of fat accumulation.

[0120] 〔Fifth Test〕 By adding the biological function regulators of Sample 1 and Sample 2, and Sample 104 to human normal epidermal keratinocytes NHEK (NB) (catalog number: "KK4009", Lot. 05298, manufactured by Kurabo Industries Ltd.), it was tested whether the biological function regulators of Sample 1 and Sample 2 have an epidermal metabolism promoting effect. Specifically, the fifth test was performed according to the following procedure.

[0121] <Test Method> Human normal epidermal keratinocytes NHEK (NB) (manufactured by Kurabo Industries Ltd.) were pre-cultured with a culture medium (product name: "HuMedia KG2", manufactured by Kurabo Industries Ltd.). Then, from the pre-cultured medium and the above cells, a cell suspension with the above cells at 1.5×10 4 cells / mL was prepared, and 3×10 4 cells of the above cell suspension were seeded in each well of a 6-well plate (catalog number: "353046", manufactured by Falcon) and cultured for 4 days. Next, after confirming that the above cells had reached 90% confluence in the above plate, the medium in the petri dish was replaced with a test medium (product name: "HuMedia KB2", manufactured by Kurabo Industries Ltd.). Furthermore, the biological function regulators of Sample 1 and Sample 2, Sample 104, and RO water were added to each well of each plate so that the final concentration was 0.1% by mass, and the cells were cultured under the conditions of 37°C (5% CO2). Here, the cells in the above plate for measuring the gene expression level of keratin 10 (KRT10) described later were cultured for 24 hours, and involucrin (Ivl) and transglutaminase described later 1The cells in the plate for measuring the gene expression level of (TGM1) were cultured for 48 hours.

[0122] Thereafter, each well was washed twice with PBS, 1 mL of TRIzol reagent (Catalog number: "15506-026", manufactured by Thermo Fisher Scientific) was added to each well, and after 1 minute, a scraper was used to collect all the cells in each well into a 1.5 mL centrifuge tube. Then, total RNA was extracted from the above cells according to the TRIzol protocol, and after measuring the absorbance at 260 nm, it was adjusted to a concentration of 1 μg / mL of total RNA. The purity of the total RNA was calculated by A260 / A280, and those showing a value of 1.8 or more were used. cDNA was obtained by reverse transcribing the above total RNA using the High Capacity RNA to cDNA Kit (Catalog number: "4387406", manufactured by Life Technologies), and the cDNA was subjected to real-time RT-PCR.

[0123] In the above real-time RT-PCR, the mRNA amounts of KRT10 (primer: Hs01043114_gl, manufactured by Thermo Fisher Scientific), TGM1 (primer: Hs01070310_ml, manufactured by Thermo Fisher Scientific), and Ivl (primer: Hs00846307_sl, manufactured by Thermo Fisher Scientific) were measured for each sample as target genes. As the internal standard (reference gene), GAPDH (catalog number: "4352934E", manufactured by Thermo Fisher Scientific) was used. The calculation used the calibration curve method. As the primer and probe, FAM dye was used. The real-time RT-PCR was performed using an apparatus (product name: "Step One Plus", manufactured by Applied Biosystems), and a reagent kit (product name: "TaqMan (registered trademark) fast advanced master mix", catalog number: "4444556", manufactured by Applied Biosystems) was used. The PCR conditions were set to perform 40 cycles of initial denaturation (95°C, 20 seconds, 1 cycle), annealing (95°C, 1 second), and polymerization (60°C, 20 seconds). Here, the total RNA (cDNA) obtained from the well added with RO water becomes Blank. The results are shown in Tables 6 to 8.

[0124] Table 6 shows the relative values (average value (Ave) and standard deviation of 3 samples each) of the gene expression levels of KRT10 in Sample 1, Sample 2, and Sample 104 relative to Blank. Table 7 shows the relative values (average value (Ave) and standard deviation of 3 samples each) of the gene expression levels of TGM1 in Sample 2 and Sample 104 relative to Blank. Table 8 shows the relative values (average value (Ave) and standard deviation of 3 samples each) of the gene expression levels of Ivl in Sample 1, Sample 2, and Sample 104 relative to Blank. Furthermore, Tables 6 to 8 also show the significant differences regarding whether the expression of each gene determined by performing statistical processing on each measured value is enhanced or not. For the above statistical processing, analysis software (product name: "Excel", manufactured by Microsoft Corporation) was used. Regarding the above significant differences, Paired-T-test was performed for determination, and p < 0.001 was represented by ***, p < 0.01 was represented by **, and p < 0.05 was represented by *.

[0125]

Table 6

[0126]

Table 7

[0127]

Table 8

[0128] <Discussion> According to Tables 6 to 8, the biological function regulators of Sample 1 and Sample 2 can enhance the gene expression levels of KRT10, TGM1, and Ivl in human epidermal cells. Thus, it is suggested that the metabolism in the epidermis is promoted by the enhanced expression of the above genes.

[0129] 〔Sixth Test〕 By conducting the above-described fifth test using the biological function regulators of Samples 41 to 49 described above, it was tested whether the biological function regulators of Samples 41 to 49 have an epidermal metabolism promoting effect. The results are shown in Tables 9 to 11.

[0130] Table 9 shows the relative values (average values (Ave) and standard deviations of three specimens each) of the gene expression levels of KRT10 in Samples 41 to 49 with respect to Blank. Table 10 shows the relative values (average values (Ave) and standard deviations of three specimens each) of the gene expression levels of TGM1 in Samples 41 to 49 with respect to Blank. Table 11 shows the relative values (average values (Ave) and standard deviations of three specimens each) of the gene expression levels of Ivl in Samples 41 to 49 with respect to Blank. In the sixth test, no determination was made regarding the presence or absence of a significant difference in the epidermal metabolism promoting effect (enhanced expression of the above-described genes).

[0131]

Table 9

[0132]

Table 10

[0133]

Table 11

[0134] <Discussion> According to Tables 9 to 11, the biological function regulators of Samples 41 to 49 can enhance the expression levels of the genes of KRT10, TGM1, and Ivl in human epidermal cells. Thus, it is suggested that the metabolism in the epidermis is promoted by the enhanced expression of the above-described genes.

[0135] 〔Seventh Test〕 In the same manner as in the first test, a mouse was administered with the biological function regulator of Sample 2, and a test was conducted using the change in the body weight of the mouse as an index to determine whether the biological function regulator has an action of suppressing fat accumulation. However, for the FCP group, the concentration of Sample 2 to be ingested was set to half the amount of the first test (feed containing 2.5% by mass).

[0136] <Discussion> Figure 12 is a graph showing the change in the body weight of mice in each group in the seventh test. According to Figure 12, it is understood that the increase in body weight of the FCP group is significantly suppressed compared to the H group.

[0137] [Eighth Test] By administering the biological function regulators of Sample 3 and Sample 4 to mice made obese by glucose loading, a test was conducted to determine whether the biological function regulators have an action of suppressing fat accumulation. Specifically, the eighth test was carried out according to the following procedure.

[0138] <Test Method> Forty-eight 5-week-old male C57BL / 6J mice were prepared by purchasing them from CLEA Japan, Inc. The body weights of the above mice were intentionally increased by allowing them to ingest normal chow (AIN-93G purified diet) and water containing 15% by mass of fructose (manufactured by FUJIFILM Wako Pure Chemical Corporation) (hereinafter also referred to as "15% by mass fructose water") for 42 days. Next, the 48 mice with increased body weights were divided into six groups (n = 8): a normal group (Group A) that ingested the above normal chow and water so that the body weights were constant among the groups, a negative control group (Group B) that ingested the above normal chow and 15% by mass fructose water, an FCP-3 group (Group C) that ingested a diet in which 6% of the 20% by mass casein contained in the normal chow was replaced with the biological function regulator of Sample 3 and 15% by mass fructose water, an FCP-4 group (Group D) that ingested a diet in which 6% of the 20% by mass casein contained in the normal chow was replaced with the biological function regulator of Sample 4 and 15% by mass fructose water, a Collapep PU group (Group E) that ingested a diet in which 6% of the 20% by mass casein contained in the normal chow was replaced with Sample 101 and 15% by mass fructose water, and an SCP-5200 group (Group F) that ingested a diet in which 6% of the 20% by mass casein contained in the normal chow was replaced with Sample 104 and 15% by mass fructose water.

[0139] Thereafter, the above mice were bred (pair-feeding) by feeding the corresponding diet and water to the mice in each group for 35 days. During the breeding period, the feed intake and body weight of each mouse were measured daily at a predetermined time. Table 12 shows a list of the diets (unit: mass%) and moisture given to the mice in each group. In the item "Water / 15 mass% fructose water" in Table 12, "W" indicates that water was given, and "F" indicates that fructose water was given. Here, the composition of the normal diet (AIN-93G purified feed) is 20 mass% casein, 26.75 mass% corn starch, 10 mass% sucrose, 20 mass% corn oil, 13.2 mass% α-corn starch, 5 mass% cellulose, 0.25 mass% choline bitartrate, 3.5 mass% mineral mix (AIN-93), 1 mass% vitamin mix (AIN-93G), and 0.3 mass% L-cystine.

[0140] [Table 12]

[0141] Next, after anesthetizing the above mice in each group with isoflurane, blood was collected by decapitation and dissection was performed Internal organs to obtain fat, serum, and liver. As the visceral fat, mesenteric fat, perirenal fat, and peritesticular fat were excised. After measuring the fat content (mass (g)) in each organ, the total Internal organs fat content was determined by summing them up. Furthermore, from the above serum, the blood glucose level, insulin amount, and leptin concentration in the serum were quantified. The blood glucose level was measured using a Glutest sensor (manufactured by Sanko Chemical Laboratories, Inc.). The insulin amount was measured using Levis Insulin-Mouse T (manufactured by Fujifilm Wako Pure Chemical Corporation). The leptin concentration was determined according to the protocol of the Molinaga Mouse / Rat Leptin Measurement Kit (manufactured by Morinaga Institute of Biological Science, Inc.).

[0142] For the liver, the following procedure was performed. First, 0.5 g of the above liver was homogenized with 2.5 mL of a 1.15% by mass potassium chloride (KCl) solution and placed in an ice-cooled test tube. Further, the above test tube was centrifuged (gravitational acceleration 9830 G, 10 minutes, 4 °C), and the supernatant was used as the crude enzyme solution. Next, based on the methods described by Nepokroeff et al. (Methods Enzymol, 35: 37-44, 1975) and Kelley et al. (Biochem. J, 235: 87-90, 1986), the enzyme activity was measured (wavelength 340 nm) from the rate of decrease in NAPDH in the presence of 100 μM malonyl CoA and 25 μM acetyl CoA with respect to the above crude enzyme solution, and the strength of fatty acid synthase (FAS) activity was determined. Further, based on the method described by Markwell et al. (J Biol. Chem, 248: 3426-3432, 1973), the enzyme activity was measured (wavelength 412 nm) from the reaction rate of dithiobisnitrobenzoic acid (DTNB) in the presence of 2 mM palmitoyl CoA and 125 mM L-carnitine with respect to the above crude enzyme solution, and the strength of fatty acid degrading enzyme (carnitine palmitoyl transferase: CPT) activity was determined. The results are shown in FIGS. 13 to 22. Regarding the significant differences between each group (Group A to Group F) in FIGS. 13 to 22, a Tukey HSD multiple comparison test was performed, and P < 0.05 was determined to be statistically significant.

[0143] <Discussion> FIG. 13 is a graph showing the body weights of the mice in each group in the 8th test. According to FIG. 13, it is understood that Groups B, E, and F were unable to suppress the weight gain compared to Group A, while Groups C and D showed a significantly suppressed weight gain compared to Group A.

[0144] FIG. 14 is a graph showing the amount of intestinal membrane fat of the mice in each group in the 8th test. FIG. 15 is a graph showing the amount of perirenal fat of the mice in each group in the 8th test. FIG. 16 is a graph showing the amount of peritesticular fat of the mice in each group in the 8th test. FIG. 17 is a graph showing the total Internal organsIt is a graph showing the amount of fat. According to FIGS. 14 to 17, it is evaluated that the fat accumulation in each organ of Group C and Group D is significantly suppressed or tends to be suppressed compared to that of Group B, Group E, and Group F. Thus, it is suggested that Group C and Group D can obtain an effect of suppressing visceral fat accumulation under fructose loading.

[0145] FIG. 18 is a graph showing the blood glucose levels in the sera of the mice in each group in the 8th test. FIG. 19 is a graph showing the insulin amounts in the sera of the mice in each group in the 8th test. According to FIG. 18, it is suggested that the blood glucose levels of Group C and Group D are significantly decreased compared to Group B. Furthermore, according to FIG. 19, it is suggested that the insulin amounts of Group C and Group D are significantly decreased compared to Group B, Group E, and Group F.

[0146] FIG. 20 is a graph showing the leptin concentrations in the sera of the mice in each group in the 8th test. According to FIG. 20, it is suggested that Group C and Group D can obtain an effect of suppressing visceral fat accumulation even under fructose loading due to a decrease in the amount of leptin production in the living body.

[0147] FIG. 21 is a graph showing the strength of FAS activity in the livers of the mice in each group in the 8th test. According to FIG. 21, it is evaluated that the synthesis of fatty acids in Group C and Group D is significantly suppressed compared to that of Group B, Group E, and Group F. Thus, it is suggested that Group C and Group D can obtain an effect of suppressing visceral fat accumulation under fructose loading due to the suppression of fatty acid synthesis. Furthermore, FIG. 22 is a graph showing the strength of CPT activity in the livers of the mice in each group in the 8th test. According to FIG. 22, it is evaluated that the decomposition of fatty acids in Group C and Group D is significantly promoted compared to that of Group B, Group E, and Group F. Thus, Group C and Group Group D is suggested to obtain an effect of suppressing visceral fat accumulation under fructose loading due to the promotion of fatty acid decomposition.

[0148] [9th Test] The biological function regulator of Sample 3 was administered to Hos:HR-1 mice suitable for observing changes in the skin to test whether the biological function regulator has an effect of promoting skin metabolism. Specifically, the ninth test was performed according to the following procedure.

[0149] <Test method> Fifteen 7-week-old female Hos:HR-1 mice were prepared by purchasing them from Hoshino Laboratory Animal Breeding Co., Ltd., and the mice were bred until they reached 8 weeks of age. Thereafter, the mice were divided into three groups (n = 5): group X, which was allowed to ingest Lab MR Stock feed (manufactured by Nippon Nosan Kogyo Kabushiki Kaisha) and water as normal diet; group Y, which was allowed to ingest a purified feed for HR-AD and water; and group Z, which was allowed to ingest a purified feed for HR-AD supplemented with 5% by mass of the biological function regulator of Sample 3 and water. Thereafter, the mice in each group were bred for 8 weeks (56 days) by feeding them the corresponding feed and water. During the breeding period, the feed intake and body weight of each mouse were measured once a week, and photographs were taken. Furthermore, mice to be euthanized before the start of breeding and at the end of breeding were selected from the mice in each group, and the skin of the back and abdomen (lumbar region) of the euthanized mice was collected. Furthermore, HE-stained specimens were prepared for the above skin, and histopathological observations were performed, including measuring the thickness of the stratum corneum of the above skin. The thickness of the stratum corneum of the skin measured in each part of the back (nape and center of the back) and abdomen (lumbar region) of the above mice is shown in Table 13. The purified feed for HR-AD is a feed that can evaluate atopic dermatitis because atopic dermatitis (AD)-like symptoms are caused by a deficiency of polyunsaturated fatty acids (n-6 PUFAs).Its composition contains, as amino acids, 0.75% by mass of arginine, 0.60% by mass of histidine, 1.03% by mass of isoleucine, 2.02% by mass of leucine, 1.69% by mass of lysine, 0.69% by mass of methionine, 1.17% by mass of tyrosine, 0.62% by mass of alanine, 2.32% by mass of proline, 1.03% by mass of phenylalanine, 0.07% by mass of tryptophan, 1.27% by mass of valine, 0.08% by mass of cystine, 0.39% by mass of glycine, 0.87% by mass of threonine, 1.18% by mass of serine, 1.47% by mass of aspartic acid, and 4.74% by mass of glutamic acid. As vitamins, it contains 32157 IU of vitamin A, 4799 IU of vitamin D3, 160 mg of vitamin E, 5 mg of vitamin K, 868.2 mg of choline, 0.09 mg of folic acid, 320 mg of niacin, 0.8 mg of biotin, 13 mg of vitamin B1, 16.3 mg of vitamin B2, 52.7 mg of vitamin B6, 0.08 mg of vitamin B12, 129.6 mg of vitamin C, and 29.7 mg of pantothenic acid per 1 kg. As minerals, it contains 0.9% by mass of calcium, 0.33% by mass of chlorine, 0.02% by mass of magnesium, 0.77% by mass of phosphorus, 0.42% by mass of potassium, 0.2% by mass of sodium, 0.00% by mass of selenium, 0.22 mg / kg of iodine, 276.78 mg / kg of iron, 0.002% by mass of cobalt, 79.28 mg / kg of manganese, 122.52 mg / kg of zinc, and 21.50 mg / kg of copper. In Table 13, "Mean" means the median, and "S.D." means the standard deviation.

[0150]

Table 13

[0151] <Discussion> First, in Group Y, desquamation was observed on the skin of the back (the nape and the center of the back) and the abdomen (the waist), and a significant increase in the thickness of the stratum corneum was recognized. As a result, it was confirmed that the mice in Group Y were established as an atopic dermatitis-like model accompanied by skin dryness and epidermal hyperplasia. On the other hand, in Group Z, although there was no change in the skin dryness compared to Group Y, as shown in Table 13, the thickness of the stratum corneum in the nape was significantly low, and it was considered that the epidermal hyperplasia was suppressed based on the skin metabolism-promoting effect of the above biological function regulator.

[0152] Test 10 The biological function regulator of Sample 4 was administered to healthy men and women aged 20 to 70 years or younger to test whether the above biological function regulator has an effect of suppressing fat accumulation in humans. Specifically, Test 10 was carried out according to the following procedure.

[0153] <Test method> This test was carried out in consideration of the "Declaration of Helsinki (Tokyo Annotation Added Edition 2004)" and the "Ethical Guidelines for Epidemiological Research (Ministry of Education, Culture, Sports, Science and Technology and Ministry of Health, Labour and Welfare Notification No. 1 of 2004)", and in compliance with the test implementation plan. The protocol of this test was approved by the Ethics Review Committee (approval number: RCB2020-001-02) and registered with UMIN (UMIN000040736). In this test, after obtaining the consent of the subjects in writing, a randomized double-blind placebo-controlled parallel-group test was carried out. In this test, measurements were taken three times at the time of screening assignment, at the pre-intake (0 time) point, and at the point three months after the intake of the test food at 5.0 g / day in absolute dry weight from the above pre-intake point, and a one-week adjustment period was provided for each measurement period. Hereinafter, the specific test content will be described.

[0154] 1) Selection criteria for subjects According to the above protocol, the selection criteria were i) men and women aged 20 years or older and 70 years or younger, ii) healthy people with no abnormalities in the health check within one year prior to the start of the test, and iii) the initial visceral fat area was 80 cm 2The above-mentioned individuals were those who could maintain an intake rate of 80% or more over the next six months. The exclusion criteria were as follows: A) those with a history of severe diseases; B) those showing abnormalities in liver and kidney function test values during a health check; C) those with impaired cardiopulmonary function; D) those with food and drug allergies; E) those who had undergone gastrointestinal surgery; F) those whom the doctor determined to have chronic or acute infectious diseases; G) those participating in other clinical studies at the start of this trial; H) those engaging in intense sports and those on a diet; I) pregnant women; J) those whom other trial supervisors or responsible persons determined to be inappropriate.

[0155] 2) Allocation of Subjects Among the 109 individuals who consented to participate in this trial, 55 were included in the trial according to the selection criteria. The responsible trial doctor created a stratified random allocation table using random numbers while taking care to ensure that the visceral fat area at the time of screening was not unbalanced among groups, and sealed it. The above allocation table was provided only to the person in charge of test food management. The above 55 trial participants were distributed the test food described below. The subjects of blinding were all the staff of the trial implementation institutions, including the sponsor, the responsible trial doctor, the trial collaborators, and the person in charge of test food management, as well as the members of the ethics review committee. The above allocation table was opened by the person in charge of statistical analysis after the trial ended.

[0156] 3) Test Foods As test foods, Sample 101 (CP group) and Sample 4 (FCP group) were used. As the control food (PL group (placebo group)), maltodextrin (trade name: "Pine Dextrose ♯2", manufactured by Matsutani Chemical Industry Co., Ltd.) was used.

[0157] 4) Intake Method The above test foods and control food were placed in aluminum pouches and distributed so that the contents were not visible. Each food (5.0 g / day) was ingested by the trial participants (subjects) at their preferred timing for three months. When they forgot to ingest, they were instructed to drink one pack (5.0 g) within the day with a limit of one pack per day.

[0158] 5) Evaluation Items 5-1) Test Schedule Various measurements were performed before ingestion (0 time) and 3 months after ingestion (3 months), and the actual measured values at each time point and the amount of change from 0 time were used as outcomes.

[0159] 5-2) Measurement of visceral fat area For the visceral fat area, it was measured using Dualscan HDS-2000 (manufactured by Omron Corporation, medical device approval number: 22300BZX00104000). The above Dualscan HDS-2000 is a visceral fat measurement device that can calculate the visceral fat area simply and safely by the dual impedance method. By passing current from two paths, it can distinguish visceral fat and abdominal subcutaneous fat, has no risk of exposure, and can measure the visceral fat area non-invasively.

[0160] 5-3) Measurement of muscle mass The measurement of muscle mass was performed only on the subjects who consented to take face photos. The subjects were measured after being acclimated at room temperature for 20 minutes or more after men only washed their faces and women cleansed and washed their faces. The above measurement was performed using a Roboskin Analyzer (manufactured by Shibuya Kogyo Co., Ltd.). Specifically, photos of the front, right, and left of the face were taken, and pigmentation was evaluated by the algorithm of the Roboskin Analyzer.

[0161] 6) Statistical analysis For the statistical analysis, STATMATE V for Windows (registered trademark) (manufactured by Atoms Co., Ltd.) was used. All statistical analyses were performed as two-sided tests, and the significance level was set at 5% with a 95% confidence interval. For the within-group comparison before and after ingestion, the Wilcoxon Signed Ranks Test was performed. For the comparison between two groups, the FCP group and the PL group, and the CP group and the PL group, the Mann-Whitney U Test was performed. For the comparison between three groups, the FCP group and the PL group and the CP group, a one-way Anova test was performed, and further, the significance difference was calculated by performing a two-sided test of the Tukey test as a post hoc test. The change in the visceral fat area is shown in Table 14, and the results of muscle mass (evaluation of the increase or decrease in pigmentation) are shown in Table 15.

[0162]

Table 14

[0163]

Table 15

[0164] <Investigation> According to Table 14, in terms of visceral fat area, no significant difference was confirmed between the CP group and the PL group and 0time. However, a significant difference was confirmed between the FCP group and 0time. Furthermore, in men over 110 cm by hierarchical level 2 In the above groups, the FCP group had a significantly decreased visceral fat area compared to the PL group. In terms of muscle quality (evaluation of increase or decrease in pigmentation), the FCP group also significantly decreased both in terms of area and number compared to the PL group. At the same time, it also significantly decreased compared to the PL group at Δ(3 months - 0time). Also, according to Table 15, in terms of muscle quality (evaluation of increase or decrease in pigmentation), the CP group significantly decreased compared to the PL group at Δ(3 months - 0time).

[0165] Seven of the subjects were unable to come to the hospital due to unavoidable reasons at the measurement in the third month and dropped out. No adverse events occurred in the subjects other than the dropouts due to the intake of the test food. From the above, the 3 - month intake of 5.0 g / day of the biological function regulator of sample 4 is safe, and the intake of the above biological function regulator was suggested to decrease the visceral fat area in healthy men over 110 cm 2 of visceral fat area. In healthy men over 110 cm 2 The significant difference appeared in the subjects over 110 cm 2 because the visceral fat amount was larger compared to those under 110 cm. It is speculated that the intake of the above biological function regulator makes it easier for visceral fat to burn, and the difference is likely to appear in 3 months. Therefore, for the visceral fat area over 110 cm 2Even in the case of minors, it is considered possible to obtain similar effects by continuing to take the above-mentioned biological function regulator. In addition, it was suggested that the intake of the above-mentioned biological function regulator suppresses not only the visceral fat area but also skin pigmentation simultaneously.

[0166] [Test 11] By adding the biological function regulator of Sample 5 to adipocytes differentiated from mouse-derived preadipocytes (3T3-L1, passage number: 7 Passages, 13 PDL), it was tested whether the biological function regulator of Sample 5 has an effect of suppressing fat accumulation. Specifically, Test 11 was carried out according to the following procedure.

[0167] (Test method) Test 11 was carried out in the same manner as Test 3 above, except that the sample added to the above adipocytes was changed to Sample 5. The results are shown in Table 16.

[0168] [Table 16]

[0169] [Discussion] According to the results of this test and Test 3, it was confirmed that Sample 5 had an effect of suppressing fat accumulation, similar to Samples 1 and 2. From this, it was confirmed that similar effects can be obtained even with different raw materials.

[0170] [Summary] From the above, it is understood that the biological function regulators of Samples 1 to 5 and Samples 41 to 49 have an effect of suppressing fat accumulation and promoting epidermal metabolism. Furthermore, it is also suggested that Samples 1 to 5 and Samples 41 to 49 have an effect of regulating the amount of adipocytokines in the living body.

[0171] The embodiments and examples disclosed herein should be considered illustrative in all respects and not restrictive. The scope of the present invention is indicated by the scope of claims rather than the above description, and it is intended that all modifications within the meaning and scope equivalent to the scope of claims be included.

Claims

Claims 1. A fermented collagen peptide obtained by fermenting with koji containing Aspergillus sojae, wherein the fermented collagen peptide has at least one action selected from the group consisting of an action of promoting epidermal metabolism, an action of suppressing fat accumulation, an action of promoting fat decomposition, and an action of regulating the amount of adipocytokines in vivo; the fermented collagen peptide comprises a collagen peptide and at least one compound selected from the group consisting of isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional; A biological function regulator used as at least one selected from the group consisting of for promoting epidermal metabolism, for suppressing fat accumulation, for promoting fat decomposition, and for regulating the amount of adipocytokines in vivo. Claims 2. The fermented collagen peptide comprises a collagen peptide and at least three compounds selected from the group consisting of isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional, the biological function regulator according to claim 1. Claims 3. An epidermal metabolism promoter comprising the biological function regulator according to any one of claims 1 or 2. Claims 4. A fat accumulation inhibitor comprising the biological function regulator according to claim 1 or 2. Claims 5. An adiponectin production promoter comprising the biological function regulator according to claim 1 or 2. Claims 6. A fat decomposition promoter comprising the biological function regulator according to claim 1 or 2. Claims 7. Comprising the biological function regulator according to claim 1 or 2, A functional food used as at least one selected from the group consisting of for promoting epidermal metabolism, for suppressing fat accumulation, for promoting fat decomposition, and for regulating the amount of adipocytokines in vivo. Claims 8. Comprising the biological function regulator according to any one of claims 1 or 2, A cosmetic used as at least one selected from the group consisting of for promoting epidermal metabolism, for suppressing fat accumulation, for promoting fat decomposition, and for regulating the amount of adipocytokines in vivo. Claims 9. A method for producing a biological function regulator containing a fermented collagen peptide, comprising: a step of preparing koji containing koji mold and a collagen raw material; a step of obtaining a biological function regulator containing the fermented collagen peptide by fermenting the collagen raw material with the koji, wherein the koji is koji containing Aspergillus sojae, The collagen raw material is at least one selected from the group consisting of the following Group 1 to Group 6, collagen extracted from at least one selected from the group, gelatin obtained by treating the collagen, and at least any one of gelatin hydrolyzates obtained by hydrolyzing the gelatin. The fermented collagen peptide is a collagen peptide and contains at least one compound selected from the group consisting of isovaleraldehyde, 1-octen-3-ol, phenylacetaldehyde, and methional. The method for producing a biological function regulator is used as at least one selected from the group consisting of promoting epidermal metabolism, suppressing fat accumulation, promoting fat decomposition, and regulating the amount of adipocytokines in the living body. Group 1: The group consisting of cowhide, skin, bone, cartilage, and tendon Group 2: The group consisting of pigskin, skin, bone, cartilage, and tendon Group 3: The group consisting of sheepskin, skin, bone, cartilage, and tendon Group 4: The group consisting of chicken skin, skin, bone, cartilage, and tendon Group 5: The group consisting of ostrich skin, skin, bone, cartilage, and tendon Group 6: The group consisting of fish bone, skin, and scale

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