Fermented product derived from plant of genus elatostema involucratum and manufacturing method thereof

Abstract

To provide a novel cosmetic containing an advanced glycation endproduct decomposition agent that effectively enhances anti-glycation effects from both inside and outside the skin.SOLUTION: Provided are a fermented product of Elatostema involucratum propagule characterized by being produced by acting koji mold on Elatostema involucratum propagule, and a cosmetic containing the fermented product of Elatostema involucratum propagule.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a fermentation product derived from a plant of the genus Eleocharis, an agent for promoting the decomposition and production of advanced glycation end products and an agent for enhancing melatonin receptor gene expression comprising the fermentation product, a composition containing the fermentation product, a skin elasticity improving agent, a skin pigmentation improving agent, and a cosmetic comprising the composition, and a method for producing the fermentation product derived from a plant of the genus Eleocharis. [Background technology]

[0002] Elatostema involucratum (also known as Mizu, Mizuna, and Akamizu) is a perennial plant classified in the Elatostema genus of the Urticaceae family, and is distributed throughout Japan, growing naturally in moist mountain forests and along mountain streams. Plants of the Elatostema genus, including Elatostema involucratum, or extracts thereof are known to have the effect of inhibiting the production of advanced glycation end products (AGEs), which are formed by the reaction of protein and sugar, and to improve wrinkles and dullness through this effect (Patent Document 1). However, they do not have the effect of breaking down AGEs that have already been produced.

[0003] AGEs is a general term for products produced in the later stages of protein glycation reactions. In recent years, it has been reported that the accumulation of AGEs is involved in the onset of various age-related diseases such as diabetes, Alzheimer's disease, cancer, and hypertension (Non-Patent Document 1). AGEs are not only contained in food, but are also known to be produced in the body by ingesting foods that contain a lot of carbohydrates (Non-Patent Documents 1, 2, 3). The accumulation of AGEs is known to be involved not only in diseases but also in various skin aging symptoms such as yellowing of the skin, blemishes, and reduced elasticity (Non-Patent Document 4).

[0004] In addition to the above-mentioned plants of the genus Sanguinea, other known ingredients that inhibit the production of AGEs include, for example, a composition containing koji-fermented ginger and an extract from the peel of the legume Dialium indum (Patent Documents 2 and 3). However, like the effect of the above-mentioned plants of the genus Sanguinea, the effect of these ingredients in inhibiting glycation reactions is a preventive measure to inhibit the production of AGEs, and they have no effect on AGEs that have already been produced by glycation reactions.

[0005] On the other hand, extracts of banaba, Meigui, etc. are known to have the effect of decomposing generated AGEs (Patent Document 4). However, all of these effects have only been evaluated in vitro, and the actual in vivo effects on the skin have not been verified. In general, human skin has a high barrier function derived from the stratum corneum, so that the penetration of ingredients applied to the skin into the skin is significantly limited. Therefore, there has been a demand for the development of an ingredient that has excellent skin penetration and high anti-glycation activity. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Patent Publication 2011-195530 [Patent Document 2] Patent No. 7076062 [Patent Document 3] Patent No. 5412798 [Patent Document 4] Patent Publication No. 2015-96498 [Non-patent literature]

[0007] [Non-Patent Document 1] Yamagata Health Insurance Medical Research, No. 24, 39-45 (2021) [Non-Patent Document 2] Aichi Prefectural University School of Nursing Bulletin, Vol. 22, 9-16 (2016) [Non-Patent Document 3] Glycative Stress Research, Volume 7, Issue 1, 70-74 (2020) [Non-Patent Document 4] Journal of the Japanese Society of Cosmetic Technologists, Vol. 53, No. 2, 83-90 (2019) Summary of the Invention [Problem to be solved by the invention]

[0008] The object of the present invention is to develop a novel drug that has the effect of promoting the decomposition of and inhibiting the production of AGEs, and to provide a skin elasticity improving agent and a cosmetic composition containing the same as an active ingredient. [Means for solving the problem]

[0009] As a result of intensive research by the present inventors to achieve the above-mentioned object, it was found that a fermented product obtained by fermenting a plant of the genus Umbellifera with bacteria of the genus Aspergillus, bacteria of the genus Lactobacillus, or yeast has an effect of effectively breaking down AGEs from inside and outside the skin, in addition to the AGE production inhibitory effect that has been conventionally known for extracts of plants of the genus Umbellifera. It was also found that it is very effective to apply this fermented product as an active ingredient in cosmetics. Furthermore, it was found that the fermented product has an effect of enhancing the expression of the melatonin receptor gene.

[0010] Melatonin is a hormone that transmits time information, but also has the effect of removing free radicals and regulating sugar metabolism and bone metabolism. In recent years, it has been attracting attention as an anti-aging ingredient, and its effect of decomposing AGEs has also been reported (Comparative Physiology and Biochemistry, 2017, Vol. 34, No. 1, 2-11; 2021, Tokyo Medical and Dental University, Faculty of Liberal Arts, Research Bulletin, No. 51, 43-50). Melatonin secretion decreases with age. Melatonin receptors are present in the human skin cell membrane, and it is believed that when the expression level of melatonin receptors is increased, the sensitivity to melatonin under physiological conditions in the skin is enhanced. Based on the above new findings, the following present invention has been completed.

[0011] (1) A fermented product derived from an Elatostema plant, obtained by fermenting all or part of the plant body of the Elatostema plant using bacteria or fungi. (2) The fermented product derived from a plant of the genus Umbellifera described in (1), wherein the fermentation is performed by a fungus of the genus Aspergillus, a fungus of the genus Lactobacillus, or a yeast. (3) The fermented product derived from a plant of the genus Eleocharis according to (1) or (2), wherein the fermented product is a solid fermented product obtained by fermenting a solid matter of the plant body, and / or a liquid fermented product obtained by fermenting an extract of the plant body. (4) An AGEs decomposition promoter and AGEs production inhibitor comprising a fermentation product derived from a plant of the genus Polytrichum according to any one of (1) to (3). (5) A melatonin receptor gene expression enhancer comprising a fermented product derived from a plant of the genus Eleocharis according to any one of (1) to (3). (6) A composition comprising a fermented product derived from a plant of the genus Eleocharis as described in any one of (1) to (3) as an active ingredient. (7) A skin elasticity improving agent comprising the composition according to (6). (8) A skin pigmentation improving agent comprising the composition described in (6). (9) A cosmetic comprising the composition according to (6). (10) A method for producing a fermented product derived from a plant of the genus Elatior, comprising: a first inoculation step of inoculating all or a part of the plant body of a plant of the genus Elatior with bacteria or fungi; a first fermentation step of fermenting the plant body after the first inoculation step under culture conditions for the bacteria or fungi to obtain a solid fermented product; a second inoculation step of mixing a suspension and / or an extract of all or a part of the plant body of a plant of the genus Elatior with the solid fermented product; and a second fermentation step of fermenting the mixed liquid obtained after the second inoculation step under culture conditions for the bacteria to obtain a liquid fermented product. (11) The method according to (10), further comprising a plant drying step of drying the plant before the first inoculation step. (12) The method according to (10) or (11), further comprising a step of preparing a solid fermentation product by drying the liquid fermentation product obtained after the second fermentation step to obtain a solid fermentation product. Effect of the Invention

[0012] The fermentation product derived from a plant of the genus Eleocharis of the present invention can provide an agent for promoting the decomposition of AGEs and inhibiting their production, as well as an agent for enhancing the expression of a melatonin receptor gene.

[0013] According to the composition of the present invention containing the fermented product derived from a plant of the genus Eleocharis, it is possible to provide a skin elasticity improving agent, a skin pigmentation improving agent, and a cosmetic composition that have the effect of promoting the decomposition and inhibiting the production of AGEs.

[0014] According to the method for producing a fermentation product derived from a plant of the genus Ulmus of the present invention, it is possible to produce a fermentation product derived from a plant of the genus Ulmus that has the effect of promoting the decomposition of AGEs and the effect of inhibiting the production of AGEs. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] The present invention will be described in detail below. 1. Fermented products derived from plants of the genus Polytrichum 1-1. Overview A first aspect of the present invention is a fermentation product derived from a plant of the genus Eleocharis. The fermentation product derived from a plant of the genus Eleocharis of the present invention is characterized by having an effect of promoting the decomposition and inhibiting the production of advanced glycation end products, and an effect of enhancing the expression of a melatonin receptor gene. The fermentation product derived from a plant of the genus Eleocharis of the present invention can provide an inhibitor of promoting the decomposition and inhibiting the production of advanced glycation end products, and an agent for enhancing the expression of a melatonin receptor gene.

[0016] 1-2.Definition of Terms As used herein, the following terms are defined. As described above, the term "Elatostema plants" as used herein refers to plants classified in the genus Elatostema of the Urticaceae family. There is no restriction on the type of plant as long as it belongs to the same genus. For example, Elatostema involucratum, E. backeri, E. fengshanense, E. lineolatum, E. malipoense, E. pleiophlebium, E. pulchra, E. repens, and E. rugosum are examples of such plants, but any of them may be used.

[0017] In this specification, the term "plant body" refers to a plant body that constitutes a plant. It is not limited to the entire plant body, but also includes parts of the plant body such as plant cells, tissues, and organs. The parts of the plant body include shoots (including leaves, stems, terminal buds, and axillary buds), flowers, roots, rhizomes, fruits, seeds, and bulbils (propagules).

[0018] As used herein, the term "substance derived from a plant of the genus Lentinus" refers to a substance derived from a plant of the genus Lentinus. Specifically, the term refers to the whole or part of the plant of a plant of the genus Lentinus, and / or an extract obtained therefrom.

[0019] As used herein, "fermentation" refers to a process in which microorganisms metabolize organic matter anaerobically or aerobically to produce metabolic products that are beneficial to humans. Fermentation as used herein is primarily, but not limited to, aerobic fermentation.

[0020] In this specification, the term "fermented product" refers to a liquid and / or solid metabolic product obtained by fermenting a target substance. In addition to the metabolic product itself obtained after fermentation, it also includes substances obtained by processing the metabolic product. In this specification, the target substance corresponds to a product derived from a plant of the genus Elatior. As described below, in this specification, "fermented product derived from a plant of the genus Elatior" may be abbreviated to simply "fermented product."

[0021] In this specification, the term "solid fermentation product" refers to a solid metabolic product obtained by solid fermentation of a solid material derived from a plant of the genus Eleocharis, and a solid material obtained by removing liquid components such as water from a liquid fermentation product described below. The term "solid material" used here refers to a solid substance that can be recognized by the naked eye.

[0022] As used herein, the term "liquid fermentation product" refers to a liquid derived from a plant of the genus Polytrichum, preferably a liquid metabolic product obtained by liquid fermentation of an extract of the plant body, and a liquid obtained by suspending the solid fermentation product in a solvent and removing the main solid components.

[0023] In this specification, the term "extract" refers to a liquid containing components of the plant body of a plant of the genus Eleocharis, including a crushed liquid obtained by crushing the plant body, a squeezed liquid obtained by squeezing the plant body, a soaked liquid obtained by soaking the plant body in a solvent, and the like, as well as a filtrate obtained by filtering these liquids and a concentrated liquid obtained by concentrating them.

[0024] In this specification, the term "microorganism" refers to bacteria or fungi that perform the fermentation. Fungi include ascomycetes, imperfect fungi, and basidiomycetes. In this specification, "bacteria or fungi" are often collectively referred to simply as "fungi."

[0025] "Advanced glycation end products" (abbreviated as "AGEs" or "AGE" in this specification) are substances that are generated when glycation products, which are the combination of protein and sugar, are heated. AGEs are generated in the body when glycation products are heated by body temperature, as well as when glycation products are heated during cooking.

[0026] In this specification, the term "drying" refers to reducing water content by evaporation or sublimation.

[0027] As used herein, the term "moisture content" refers to the weight ratio of the water content to the dry weight of a target object (mainly the plant body of a plant of the genus Alpinia in this specification).

[0028] As used herein, the term "effective amount" refers to an amount of an active ingredient that is necessary for the active ingredient to exert its function and that causes little or no harmful side effects in the living body to which it is applied. This effective amount may vary depending on various conditions such as information about the subject, the route of administration, and the number of administrations.

[0029] As used herein, the term "subject" refers to a living body to which a drug or composition is applied. Examples of such a body include humans, livestock, racehorses, pets, and laboratory animals. Preferably, the body is a human (in this case, the body is specifically referred to as a "subject").

[0030] As used herein, "information on a subject" refers to various individual information on the living body to which the invention is applied, and in the case of a subject, includes the overall health condition, the progression and severity of a disease or injury if present, age, weight, sex, diet, drug sensitivity, the presence or absence of concomitant medication, and resistance to treatment.

[0031] As used herein, the term "skin elasticity improving agent" refers to a composition that improves or restores skin elasticity lost due to AGEs, or delays the progression of loss of skin elasticity due to aging.

[0032] As used herein, the term "cosmetic" refers to the main ingredient contained in a cosmetic product.

[0033] 1-3. Composition of fermented products derived from plants of the genus Polytrichum The fermented product derived from a plant of the genus Eleocharis of the present invention (often abbreviated as simply "fermented product" in this specification) comprises metabolic products obtained by fermenting a material derived from a plant of the genus Eleocharis with a microorganism.

[0034] In the fermented product of the present invention, the material derived from a plant of the genus Poria to be used for fermentation is not particularly limited as long as it is a plant of the genus Poria. It may be the whole plant or a part of it. In the case of a part of the plant, any part of the plant may be used, and it may be a shoot, a flower, a root, a rhizome, a fruit, a seed, a bulbil, or a combination thereof.

[0035] The microorganisms used for fermentation are not particularly limited, but preferred are bacteria of the genus Lactobacillus or Bacillus, ascomycetes of Saccharomyces cerevisiae or Schizosaccharomyces pombe, and imperfect fungi of the genus Aspergillus or Monascus, commonly known as koji mold.

[0036] The type of Lactobacillus is not particularly limited, but is preferably a species that is industrially used in lactic acid fermentation, etc. Examples include Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus curvatus, Lactobacillus sakei, Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus helveticus, Lactobacillus salivarius, Lactobacillus brevis, and Lactobacillus buchneri.

[0037] The types of Aspergillus and Monascus are not particularly limited, but are preferably those called black koji mold, white koji mold, yellow koji mold, red koji mold, etc. and used industrially. For example, specific examples of black koji mold include Aspergillus awamori (black koji mold), Aspergillus saitoi (A. saitoi), Aspergillus nakazawai (A. nakazawai), Aspergillus usamii (A. usamii), Aspergillus luchensis (A. luchensis), and Aspergillus niger (A. niger). Examples of white koji mold include Aspergillus kawachii (A. kawachii). Examples of yellow koji mold include Aspergillus oryzae (A. oryzae). Examples of Red Aspergillus include Monascus purpureus, Monascus rubber, Monascus pilosus, and Monascus pubigerus.

[0038] The state of the fermented product of the present invention may be any of a solid fermented product, a liquid fermented product, or a combination thereof. The solid fermented product may be a solid product containing the plant body of a plant derived from the genus Eleocharis, or may be a dry solid product obtained by drying the liquid fermented product.

[0039] The fermented product of the present invention may be either a single-stage fermented product or a multi-stage fermented product. In this specification, the term "single-stage fermented product" refers to a fermented product obtained by only a single fermentation step on a plant derived from the genus Eleocharis. In addition, in this specification, the term "multi-stage fermented product" refers to a fermented product obtained after multiple fermentation steps on a plant derived from the genus Eleocharis. The number of times referred to here is not important. For example, two, three, four, or five times may be mentioned, but is not limited thereto. The multi-stage fermented product may be a fermented product that has been subjected to the same fermentation step in each step, or may be a fermented product that has been subjected to different fermentation steps. Examples of fermented products that have undergone different fermentation processes include secondary fermented products obtained by subjecting a material derived from a plant of the genus Sanguinea to solid fermentation as a primary fermentation, followed by liquid fermentation of a suspension of the solid fermented product in a solvent or the like; fermented products obtained by fermenting with Lactobacillus bacteria and then further fermenting with Aspergillus bacteria; and fermented products obtained by adding new material derived from a plant of the genus Sanguinea in multiple fermentation processes and then fermenting the resulting mixture.

[0040] 1-4. Use of fermented products derived from plants of the genus Alpinia As shown in the Examples below, the inventors' research has revealed that the fermented product derived from a plant of the genus Eleocharis of the present invention has an AGEs decomposition promoting effect and an effect of enhancing melatonin receptor gene expression in addition to the AGEs production inhibiting effect known for plants or extracts of plants of the genus Eleocharis. Based on these research results, the fermented product derived from a plant of the genus Eleocharis of the present invention can be used as an AGEs production inhibitor, an AGEs decomposition promoter, and an agent for enhancing melatonin receptor gene expression.

[0041] Furthermore, by utilizing these effects, the composition can be used as a cosmetic material or a skin elasticity improving agent, either as it is or in combination with other components. These will be described in detail in the third aspect.

[0042] 2. Method for producing fermented product derived from plants of the genus Polytrichum 2-1. Overview A second aspect of the present invention is a method for producing a fermentation product derived from a plant of the genus Eleocharis. The production method of the present invention produces a multi-stage fermentation product among fermentation products derived from a plant of the genus Eleocharis. According to the production method of the present invention, a fermentation product having high effects of promoting the decomposition and inhibiting the production of advanced glycation end products, and enhancing the expression of the melatonin receptor gene can be produced.

[0043] 2-2. Method The production method of the present invention includes the first inoculation step, the first fermentation step, the second inoculation step, and the second fermentation step as essential steps, and includes the grass drying step and the solid fermentation product preparation step as optional steps. Each step will be specifically described below.

[0044] (1) Plant drying process The "plant drying step" is a step of drying all or part of the plant of the genus Poria prior to the first inoculation step. This step is an optional step that is carried out prior to the first inoculation step, and may be carried out as necessary. This step reduces the moisture content in the plant of the genus Poria, thereby enabling fermentation in the first fermentation step described below to be carried out efficiently.

[0045] The plant body of the plant of the genus Alpinia used in the present invention may be the whole plant body or a part thereof. In the case of a part thereof, the part may be any part of the plant body, such as an above-ground part such as a shoot, a root, a bulbil, or a combination thereof.

[0046] The method for drying the plant body in this step is not limited as long as it can reduce the moisture content in the plant body. Examples include sun drying, dehumidification drying, hot air drying, heat drying, reduced pressure drying, freeze drying, and combinations thereof.

[0047] This step may be performed until the moisture content of the plant mass reaches a level at which bacteria or fungi can efficiently ferment the plant mass in the first fermentation step described below, for example, but not limited to, until the moisture content reaches 70% or less, 60% or less, 50% or less, or 40% or less. Therefore, the treatment time of this step may be appropriately determined depending on the moisture content of the plant mass and / or the drying method.

[0048] (2) First inoculation process The "first inoculation step" is a step of inoculating bacteria or fungi into the plant body of a plant of the genus Polytrichum to be fermented.

[0049] The type of bacteria or fungi used for fermentation in this step is not limited. For example, bacteria such as Lactobacillus or Bacillus can be preferably used. Furthermore, fungi such as Aspergillus or Monascus can be preferably used.

[0050] When inoculating, it is desirable to sterilize the plant body to be used in advance to remove any unwanted bacteria adhering to the plant body. In addition, it is preferable to inoculate in a sterile environment.

[0051] The method of inoculation is not particularly limited as long as it is a method that brings the bacteria or fungus into contact with the plant body. For example, it may be a method of immersing the plant body in a bacterial or fungal culture solution, a method of applying or spraying the culture solution or spores to the plant body, a method of adding a seed fungus to the plant body, etc. In order to prepare the culture solution of the bacteria or fungus used in this step, it may be pre-cultured.

[0052] (3) First fermentation process The "first fermentation step" is a step of fermenting the plant body after the first inoculation step under culture conditions for the bacteria or fungus to obtain a solid fermentation product.

[0053] The purpose of this step is to carry out solid fermentation to ferment the plant body, i.e., the solid matter, derived from a plant of the genus Alpinia, and to obtain a solid fermentation product as a metabolic product thereof.

[0054] The culture conditions in this step may be optimal for the bacteria or fungi used in the fermentation, and in principle, the culture may be performed under the gas phase under the optimal conditions for the bacteria or fungi to be used in the fermentation. For example, when Aspergillus bacteria are inoculated, they may be cultured for about 1 to 7 days under aeration conditions at 30°C ± 2°C in an environment free from contamination by other bacteria. When Lactobacillus bacteria are inoculated, they may be cultured for about 1 to 3 days under aeration conditions at 37°C ± 2°C.

[0055] (4)Second inoculation process The "second inoculation step" is a step of mixing the liquid-derived material of a plant of the genus Alpinia with the solid fermentation product, which is the primary fermentation product obtained in the first fermentation step. The purpose of this step is to carry out secondary fermentation by liquid fermentation on the solid fermentation product obtained in the first fermentation step.

[0056] The liquid-derived material from a plant of the genus Sanguinea may be a suspension and / or an extract containing all or a part of the plant body of a plant of the genus Sanguinea. The amount of the liquid-derived material to be added is not particularly limited. For example, the weight ratio of the liquid-derived material to the solid fermentation product obtained in the first fermentation step may be 100% to 6000%, 200% to 5000%, 300% to 4000%, 400% to 3000%, 500% to 2000%, or 800% to 1000%. Since the secondary fermentation of the solid fermentation product, the primary fermentation product, is carried out under liquid, the liquid-derived material is further added so that the secondary fermentation proceeds appropriately.

[0057] The solid fermentate may be in the form of chunks, crushed pieces, granules, powder, or a combination thereof.

[0058] The solid fermentation product used in this step can be the solid fermentation product obtained in the first fermentation step, which is used as it is, and the bacteria or fungi attached to the solid fermentation product can be inoculated as seed bacteria. If necessary, new bacteria may be added. In this case, it is preferable that the newly added bacteria is of the same species, preferably the same strain, as the bacteria used in the first inoculation step. On the other hand, if the solid fermentation product obtained in the first fermentation step has been sterilized by heating or the like before this step, the solid fermentation product does not serve as seed bacteria, and new bacteria are inoculated. In this case, too, it is preferable that the newly added bacteria is of the same species, preferably the same strain, as the bacteria used in the first inoculation step.

[0059] (5) Second fermentation process The "second fermentation step" is a step in which the mixed liquid obtained after the second inoculation step is fermented under culture conditions for the bacteria to obtain a liquid fermentation product.

[0060] In this step, the solid fermentation product, which is the primary fermentation product obtained in the first fermentation step, is subjected to liquid fermentation as a secondary fermentation, with the aim of obtaining a liquid fermentation product as a metabolic product thereof.

[0061] The culture conditions in this step are basically the same as those in the first fermentation step, except that the liquid fermentation is used. The first fermentation step was carried out in a gas phase, but this step is carried out in a liquid phase. Although not limited thereto, the fermentation in the present invention is mainly aerobic fermentation, and in the case of aerobic fermentation, it is preferable to subject the mixed liquid to an aeration treatment such as stirring.

[0062] The culture liquid obtained after this step is a liquid fermented product that has undergone secondary fermentation by liquid fermentation after solid fermentation. This liquid fermented product is the fermented product derived from a plant of the genus Poria in the first embodiment as is. If necessary, the liquid fermented product may be filtered or centrifuged to remove unnecessary solid fermented product and impurities remaining in the liquid.

[0063] (6) Solid fermentation product preparation process The "solid fermentation product preparation step" is a step of drying the liquid fermentation product obtained after the second fermentation step to obtain a solid fermentation product.

[0064] In this step, the liquid fermentation product obtained in the second fermentation step as the secondary fermentation product is converted into a solid fermentation product. To convert the liquid fermentation product into a solid fermentation product, the water content can be reduced until only solid components remain. There is no limitation on the method for reducing the water content, but drying or centrifugation is generally convenient. When performing drying on the liquid fermentation product, there is no limitation on the drying method. For example, the drying method used in the plant drying step can also be used in this step. The freeze-drying method is particularly convenient for obtaining a solid fermentation product easily and relatively inexpensively without denaturing the components of the fermentation product.

[0065] Before carrying out this step, the liquid fermentation product obtained after the second fermentation step may be filtered or centrifuged in advance to remove unnecessary solid fermentation products and impurities remaining in the liquid.

[0066] The solid matter obtained after this step is a solid fermentation product that contains a secondary fermentation product that has undergone liquid fermentation after solid fermentation. This solid fermentation product is the fermentation product derived from a plant of the genus Polytrichum according to the first embodiment.

[0067] 3. Composition containing fermented product derived from a plant of the genus Alpinia 3-1. Overview The third aspect of the present invention is a composition. The composition of the present invention is characterized in that it contains the fermented product derived from a plant of the genus Eleocharis described in the first aspect as an active ingredient. This composition can be used as a skin elasticity improving agent, skin pigmentation improving agent, or cosmetic agent by utilizing the effect of inhibiting AGEs production, the effect of promoting AGEs decomposition, or the effect of enhancing melatonin receptor gene expression of the fermented product.

[0068] 3-2.Configuration 3-2-1. Constituent factors The composition of this embodiment contains an active ingredient as an essential component and a carrier and / or a solvent as optional components. Each of the components will be specifically described below.

[0069] (1) Active ingredient The composition comprises the fermented product according to the first aspect as an active ingredient. The composition may comprise two or more different fermented products. For example, the composition may comprise a fermented product fermented with Aspergillus and a fermented product fermented with Lactobacillus, or a fermented product obtained by a single fermentation and a fermented product obtained by a two-stage fermentation.

[0070] The content of the fermented product in the composition of the present invention varies depending on the type of fermented product, the formulation of the composition, the type of carrier or solvent, and the intended use, and may be appropriately determined taking into account each of the conditions.

[0071] The effective amount of the fermented product in the present composition and the application amount calculated based thereon may be determined based on information about each individual subject, etc.

[0072] (2) Solvent The composition may contain a pharma- ceutically acceptable solvent. The solvent is an optional component of the composition and may be added as necessary. "Pharmaceutically acceptable" means that the solvent is harmless or has low toxicity to the living body, can be generally used in the technical field of formulations, and can preferably be used in pharmaceutical compositions.

[0073] Examples of the solvent include water or an aqueous solution, or an organic solvent. Examples of the pharma- ceutically acceptable aqueous solution include physiological saline, an isotonic solution containing glucose or other auxiliary agents, a phosphate buffer, and a sodium acetate buffer. Examples of the auxiliary agent include D-sorbitol, D-mannose, D-mannitol, sodium chloride, and other low-concentration nonionic surfactants, polyoxyethylene sorbitan fatty acid esters, etc. Examples of the pharma-ceutically acceptable organic solvent include ethanol, butanol, etc.

[0074] (3) Carrier The composition may include a pharma- ceutically acceptable carrier, which is an optional component of the composition and may be added if desired.

[0075] Examples of the carrier include suspending agents, diluents, solubilizing agents, dispersing agents, surfactants, emulsifying agents, soothing agents, stabilizers, preservatives, antiseptics, antioxidants, buffers, and isotonic agents.

[0076] In addition to the above, if necessary, the composition may appropriately contain excipients, fillers, binders, disintegrants, absorption enhancers, bulking agents, moisturizing agents, humectants, adsorbents, disintegration inhibitors, coating agents, colorants, and the like that are commonly used in pharmaceuticals.

[0077] Such carriers are primarily used to facilitate the formation of dosage forms and maintain the dosage form and pharmaceutical effects, as well as to make the active ingredient, the fermented product, less susceptible to decomposition in the body, and may be used appropriately as needed.

[0078] 3-2-2. Dosage form The dosage form of the present composition is not particularly limited as long as it is a form that does not inactivate the fermented product derived from a plant of the genus Eleocharis, which is an active ingredient, and can exert the pharmacological effect of the active ingredient in the body after administration. The specific dosage form of the present composition may be appropriately selected depending on the administration method and / or prescription conditions. In general, the administration method can be broadly divided into parenteral administration and oral administration, and the present composition may be made into a dosage form suitable for each administration method.

[0079] Parenteral administration can be divided into systemic administration and local administration, and local administration can be further divided into intratissue administration, epidermal administration, transmucosal administration, and rectal administration. The composition may be in a dosage form suitable for each administration method. For example, dosage forms suitable for epidermal or transmucosal administration include liquids (including liniments, eye drops, nasal drops, and inhalants), suspensions (including emulsions and creams), powders (including nasal drops and inhalants), pastes, gels, ointments, plasters, etc. Dosage forms suitable for systemic or intratissue administration include injections, which are liquids. Dosage forms suitable for rectal administration include suppositories, etc.

[0080] In the case of oral administration, the dosage form includes solid agents (including tablets, pills, sublingual agents, capsules, and drops), granules, powders, powders, liquid agents (including oral solutions, suspensions, emulsions, and syrups), etc. The solid agents can be made into dosage forms coated with a coating known in the art, such as sugar-coated tablets, gelatin-coated tablets, enteric-coated tablets, film-coated tablets, double tablets, and multi-layer tablets, if necessary.

[0081] This composition can be applied for treating or preventing diseases, like the skin elasticity improving agent described below, or as a cosmetic. The application site varies depending on the target disease. In the case of this composition, although there is no limitation, epidermis is preferred in consideration of its effect. Therefore, although the administration method is not limited, epidermal administration is preferred.

[0082] The specific shape and size of each of the above dosage forms are not particularly limited as long as they are within the range of dosage forms known in the art.

[0083] 3-3.Applications The use of the composition is not limited, but it can be used as an external preparation for the skin based on the action and effect of the fermented product contained as an active ingredient. Examples of external preparations for the skin include skin elasticity improving agents, skin pigmentation improving agents, and cosmetics.

[0084] Cosmetics that can contain a cosmetic preparation consisting of the present composition include, but are not limited to, various types of cosmetics such as lotions, milky lotions, beauty serums, creams (cleansing creams, shaving creams, sun creams, sunscreen creams, eye creams, lip balms), lotions (sunscreen lotions, suntan lotions), packs, cosmetic soaps, foundations, face powders, powders, lipsticks, eyeliners, eyeshadows, mascaras, bath cosmetics, shampoos, conditioners, hair dyes, and hair cosmetics.

[0085] The amount of the present composition in a cosmetic product is not limited, but may be 0.001 to 100%, preferably 0.1 to 10%, of the total cosmetic ingredients.

[0086] The skin elasticity improving agent, skin pigmentation improving agent or cosmetic of the present invention can be produced by arbitrarily selecting and combining various components and additives suitable for the intended purpose from among those generally used in skin external preparations such as cosmetics. For example, there is no particular limitation as long as they are bases, drugs, etc. generally used in cosmetics, quasi-drugs and bath agents. EXAMPLES

[0087] The present invention will be described in more detail below with reference to examples, but the technical scope of the present invention is not limited to these examples.

[0088] Example 1: Preparation of fermented product derived from a plant of the genus Polytrichum of the present invention (the purpose) The fermented product of the present invention derived from a plant of the genus Polytrichum is prepared.

[0089] (material) In principle, the following procedures were carried out in a sterile environment unless otherwise specified. (1) Material Plants of the genus Alpinia Frozen coral bulbils were used. Fermentation bacteria Aspergillus oryzae (Shirayuri: Nihon Jozo Kogyo) and Aspergillus awamori (NBRC4033: National Institute of Technology and Evaluation) were used. Lactic acid bacteria: Lactobacillus plantarum (NBRC3074: National Institute of Technology and Evaluation) was used. Yeast: Saccharomyces cerevisiae (NBRC0216: National Institute of Technology and Evaluation) was used.

[0090] (method) (1) (Production Example 1) Preparation of Aspergillus oryzae spore suspension for inoculation The koji molds were inoculated onto potato dextrose agar plates (AS ONE Corporation) and cultured for one week at 30° C. to form plate spores. 15 mL of sterile saline was added to the plates to obtain spore suspensions for inoculation.

[0091] (2) (Production Example 2) Preparation of a single solid fermentation product of Uwabamisou with Aspergillus oryzae (Uwabamisou Aspergillus oryzae single solid fermentation product) 100 g of frozen coral bulbils were naturally thawed, then dried at 80°C in a ventilated dryer until the weight was reduced to half, then placed in a polypropylene bag for cultivation on a culture bed (ST-12P25, Shinano Poly Co., Ltd.) and sterilized in an autoclave. After cooling, 2.5 mL of the spore suspension was inoculated and cultured at 30°C for 2 to 4 days to obtain a solid fermentation product of coral bulbil.

[0092] (3) (Production Example 3) Preparation of two-stage (solid / liquid) fermentation liquid of Uwabamisou with Aspergillus oryzae (Uwabamisou Aspergillus oryzae two-stage solid / liquid fermentation liquid) A two-stage liquid fermentation liquid was prepared by the manufacturing method described in the second embodiment of the present specification. After thawing 460 g of the frozen Uwabamisou basket, 720 g of water was added and the mixture was ground until it became a paste. The entire amount of the obtained Uwabamisou basket paste was introduced into a jar fermenter (Takasugi Seisakusho), sterilized in an autoclave, and then cooled. To this was added 20 g of the solid fermentation product prepared in (2) Manufacturing Example 2 as a preculture product, and cultured at 30°C for 24 hours. After the culture was completed, the temperature was raised to 80°C and maintained for 3 hours, and then cooled to obtain a Uwabamisou two-stage solid / liquid Aspergillus fermentation liquid.

[0093] (4) (Production Example 4) Preparation of a two-stage (solid / liquid) fermentation extract of Uwabamisou with Aspergillus oryzae (Uwabamisou Aspergillus oryzae two-stage solid / liquid fermentation extract) A solid fermentation product of the two-stage liquid fermentation product was prepared by the production method described in the second aspect of the present specification. The same weight of ethanol was added to 500 g of the two-stage liquid fermentation product obtained in (3) Production Example 3 above, and extraction was carried out overnight. The obtained extract was centrifuged and further filtered under reduced pressure to obtain a filtrate. Ethanol and water were evaporated from the filtrate using an evaporator, and the obtained concentrated liquid was freeze-dried using a freeze-dryer to obtain a two-stage solid / liquid fermentation extract of Corylus orbiculatus.

[0094] (5) (Production Example 5) Preparation of a two-stage (solid / liquid) fermentation extract of Uwabamisou with Aspergillus oryzae (Uwabamisou Aspergillus oryzae two-stage solid / liquid fermentation BG extract) The basic procedure was the same as that of the extraction method in (4) Production Example 4 above, except that 1,3-butanediol (1,3-butylene glycol: BG) was used instead of ethanol extraction. The solid content was measured by the evaporation to dryness method, and the filtrate was diluted to a solid content of 0.5 mg / mL to obtain an extract of Aspergillus oryzae two-stage solid / liquid fermentation (BG extract of Aspergillus oryzae two-stage solid / liquid fermentation).

[0095] (6) (Comparative Manufacturing Example 1) Preparation of a non-fermented extract of Uwabamisima for comparison (Uwabamisima non-fermented extract) After thawing 460 g of frozen Uwabamisou basket, 720 g of water was added and the mixture was ground until it became a paste. The entire amount of the obtained Uwabamisou basket paste was sterilized in an autoclave and then cooled to obtain a hot water-treated Uwabamisou basket product. The same weight of ethanol was added to 500 g of the obtained hot water-treated Uwabamisou basket product and extracted overnight. The obtained extract was centrifuged and further filtered under reduced pressure to obtain a filtrate. Ethanol and water were evaporated using an evaporator, and the concentrated liquid was freeze-dried using a freeze dryer to obtain a non-fermented Uwabamisou basket extract.

[0096] (7) (Comparative Manufacturing Example 2) Preparation of a non-fermented extract of Uwabamisou extract for comparison (Uwabamisou non-fermented BG extract) The same weight of 1,3-butanediol was added to 500 g of the hot water treated product of Uwabamisou obtained in (6) Comparative Production Example 1, and extraction was carried out overnight. The obtained extract was centrifuged and further filtered under reduced pressure to obtain a filtrate. The solid content was then measured by the evaporation to dryness method, and the filtrate was diluted so that the solid content in the filtrate was 0.5 mg / mL to obtain a non-fermented Uwabamisou extract (non-fermented Uwabamisou BG extract).

[0097] (8) (Comparative Production Example 3) Preparation of Aspergillus oryzae preculture solution After thawing 40 g of frozen Aspergillus oryzae, 100 g of water was added and the mixture was ground to a paste. After autoclaving, the mixture was allowed to cool, and then 2.5 mL of the spore suspension prepared in (1) Production Example 1 was added. The mixture was cultured at 30°C for 48 hours with shaking to obtain a preculture solution of Aspergillus oryzae oryzae.

[0098] (9) (Comparative Production Example 4) Preparation of Aspergillus oryzae liquid fermentation solution After thawing 460 g of frozen Aspergillus oryzae, 620 g of water was added and the mixture was mashed into a paste. The entire amount of the paste was then placed in a jar fermenter (Takasugi Seisakusho) and sterilized in an autoclave, after which it was cooled. To this was added 140 g of the preculture prepared in (8) Comparative Production Example 3, and cultured at 30°C for 24 hours with stirring. After the culture was completed, the temperature was raised to 80°C and maintained for 3 hours, and then cooled to obtain Aspergillus oryzae liquid fermentation liquid.

[0099] (10) (Comparative Production Example 5) Preparation of Aspergillus oryzae Liquid Fermentation Extract (9) 500 g of the Aspergillus oryzae liquid fermentation liquid obtained in Comparative Production Example 4 was mixed with the same weight of ethanol and extracted overnight. The resulting extract was centrifuged and filtered under reduced pressure to obtain a filtrate. Then, ethanol and water were evaporated using an evaporator, and the resulting concentrated liquid was freeze-dried using a freeze dryer to obtain an Aspergillus oryzae liquid fermentation extract.

[0100] (11) (Comparative Production Example 6) Preparation of Aspergillus oryzae Liquid Fermentation Extract (Aspergillus oryzae Liquid Fermentation BG Extract) (9) 500 g of the Aspergillus oryzae liquid fermentation liquid obtained in Comparative Production Example 4 was mixed with the same weight of 1,3-butanediol and extracted overnight. The resulting extract was centrifuged and filtered under reduced pressure to obtain a filtrate. The solid content in the filtrate was then measured by the evaporation to dryness method, and the filtrate was diluted to a solid content of 0.5 mg / mL to obtain an Aspergillus oryzae liquid fermentation extract (Aspergillus oryzae liquid fermentation BG extract).

[0101] (12) (Comparative Production Example 7) Preparation of liquid fermentation solution of Uwabamisou lactic acid bacteria A loopful of Lactobacillus plantarum (NBRC3074) was inoculated into a sterilized lactic acid bacteria inoculation medium (Nissui Co., Ltd.) and cultured at 37° C. for 48 hours to prepare a preculture solution. Next, 460g of frozen Uwabamisou basket was thawed, 620g of water was added, and the mixture was ground into a paste. The entire amount of the Uwabamisou basket paste and 6g of yeast extract (Becton Dickinson) were placed in a jar fermenter (Takasugi Seisakusho), sterilized in an autoclave, and then cooled. 100g of the preculture was added and cultured at 37℃ for 48 hours with stirring. After the culture was completed, the temperature was raised to 80℃ and maintained for 3 hours, then cooled to obtain Uwabamisou lactic acid bacteria liquid fermentation liquid.

[0102] (13) (Comparative Production Example 8) Preparation of Liquid Fermentation Extract of Lactobacillus subtilis (12) 500 g of the liquid fermentation liquid of the lactic acid bacteria of Uwabamia obtained in Comparative Production Example 7 was added with the same weight of ethanol and extracted overnight. The resulting extract was centrifuged and filtered under reduced pressure to obtain a filtrate. Ethanol and water were then evaporated using an evaporator, and the resulting concentrated liquid was freeze-dried using a freeze-dryer to obtain a liquid fermentation extract of the lactic acid bacteria of Uwabamia.

[0103] (14) (Comparative Production Example 9) Preparation of Uwabamisou Lactic Acid Bacteria Liquid Fermentation Extract (Uwabamisou Lactic Acid Bacteria Liquid Fermentation BG Extract) (12) 500 g of the lactic acid bacteria of Uwabamisae liquid fermentation liquid obtained in Comparative Production Example 7 was added with the same weight of 1,3-butanediol and extracted overnight. The resulting extract was centrifuged and filtered under reduced pressure to obtain a filtrate. The solid content was then measured by an evaporation-to-dryness method, and the filtrate was diluted to a solid content of 0.5 mg / mL to obtain an extract of lactic acid bacteria of Uwabamisae liquid fermentation (BG extract of lactic acid bacteria of Uwabamisae liquid fermentation).

[0104] (15) (Comparative Production Example 10) Preparation of liquid fermentation solution of Uwabamisou yeast A loopful of Saccharomyces cerevisiae (NBRC0216) was inoculated into sterilized YM liquid medium (10 g glucose, 5 g peptone, 3 g yeast extract, and 3 g malt extract dissolved in 1 L of distilled water) and cultured with shaking at 30°C for 24 hours to prepare a preculture solution. Next, 460 g of frozen Uwabamisou basket was thawed, 620 g of water was added, and the mixture was ground into a paste. The entire amount of the Uwabamisou basket paste was placed in a jar fermenter (Takasugi Seisakusho), sterilized in an autoclave, and then cooled. 100 g of preculture was added to this, and cultured at 30°C for 24 hours with stirring. After the culture was completed, the temperature was raised to 80°C and maintained for 3 hours, and then cooled to obtain a liquid fermentation liquid of Uwabamisou yeast.

[0105] (16) (Comparative Production Example 11) Preparation of Liquid Fermentation Extract of Polygala gracilis Yeast (15) 500 g of the liquid fermentation liquid of the yeast of the Uwabamia obtained in Comparative Production Example 10 was added with the same weight of ethanol and extracted overnight. The obtained extract was centrifuged and filtered under reduced pressure to obtain a filtrate. Furthermore, ethanol and water were evaporated using an evaporator, and the obtained concentrated liquid was freeze-dried using a freeze-dryer to obtain a liquid fermentation extract of the yeast of the Uwabamia.

[0106] (17) (Comparative Production Example 12) Preparation of Uwabamisou Yeast Liquid Fermentation Extract (Uwabamisou Yeast Liquid Fermentation BG Extract) (15) 500 g of the yeast liquid fermentation liquid obtained in Comparative Production Example 10 was added with the same weight of 1,3-butanediol and extracted overnight. The resulting extract was centrifuged and filtered under reduced pressure to obtain a filtrate. The solid content was then measured by the evaporation to dryness method, and the filtrate was diluted to a solid content of 0.5 mg / mL to obtain a Uwabamisou yeast liquid fermentation extract (Uwabamisou yeast liquid fermentation BG extract).

[0107] The samples prepared in this example are summarized in Table 1.

[0108] [Table 1] TIFF2025008335000002.tif73164

[0109] <Example 2: Verification of the inhibitory effect of fermentation product derived from a plant of the genus Polytrichum commune on the formation of advanced glycation end products> (the purpose) The fermentation product derived from a plant of the genus Eleocharis of the present invention is verified to have an inhibitory effect on the production of advanced glycation end products (AGEs).

[0110] (method) In 250 mM phosphate buffer (pH 7.4), each powder of (4) the Aspergillus oryzae two-stage solid / liquid fermentation extract obtained in Production Example 4, (6) the Aspergillus oryzae non-fermented extract obtained in Comparative Production Example 1, (10) the Aspergillus oryzae liquid fermentation product obtained in Comparative Production Example 5, (13) the Aspergillus oryzae lactic acid bacteria liquid fermentation extract obtained in Comparative Production Example 8, and (16) the Aspergillus oryzae yeast fungus liquid fermentation extract obtained in Comparative Production Example 11 in Example 1 was dissolved to an arbitrary concentration to prepare each test group. Phosphate buffer was used as a control group. Furthermore, 200 μL of 2.5 mg glucose aqueous solution and 200 μL of 10 mg / mL BSA aqueous solution were added to each sample, and the mixture was reacted at 60° C. for 24 hours.

[0111] The reaction was stopped by adding 100 μL of trichloroacetic acid, and the mixture was then centrifuged to remove the supernatant. The precipitate was dissolved by adding 1000 μL of alkaline phosphate buffer. The resulting solution was measured at an excitation wavelength of 360 nM and a fluorescence wavelength of 460 nm. This fluorescence measurement is proportional to the amount of glycation products produced by the reaction of glucose with protein (BSA). The measurement value for each test group was divided by the measurement value for the control group to determine the inhibition rate of glycation product production.

[0112] (result) The results are shown in Table 2. When the non-fermented extract of Arctium arbutifolia obtained in Comparative Production Example 1 was added, a production inhibition rate of about 30% was observed at a final concentration of 0.02%, and about 50% at 0.04%. This does not contradict the results described in Patent Document 1, which shows that the plant body of Arctium arbutifolia plants has an AGE production inhibitory effect. On the other hand, when the Arctium arbutifolia Aspergillus two-stage solid / liquid fermentation extract of Arctium arbutifolia in Production Example 4 was added, a production inhibition rate of about 36-37% at a final concentration of 0.02%, and about 61-63% at 0.04%, was observed for fermentation with either A. oryzae or A. awamori Aspergillus oryzae, which were higher values ​​than those of Comparative Production Example 1, which used the non-fermented extract. Furthermore, when the liquid fermentation extract of Aspergillus oryzae obtained in Comparative Production Example 5 was added, the production inhibition rate was 32.1% at a final concentration of 0.02% and 57.9% at 0.04%, and when fermented with Aspergillus oryzae, a slightly higher production inhibition rate was observed compared to the non-fermented Comparative Production Example 1, even in the case of single fermentation. When the liquid fermentation extract of Aspergillus oryzae lactic acid bacteria obtained in Comparative Production Example 8 was added, the production inhibition rate was 30.2% at a final concentration of 0.02% and 52.4% at 0.04%, and when fermented with not only Aspergillus oryzae but also lactic acid bacteria, a slightly higher production inhibition rate was observed compared to the non-fermented Comparative Production Example 1. When the liquid fermentation extract of Aspergillus oryzae yeast fungus obtained in Comparative Production Example 11 was added, the production inhibition rate was 27.3% at a final concentration of 0.02%, which was slightly lower than that of Comparative Production Example 1, but the production inhibition rate was 53.3% at 0.04%, which was slightly higher than that of Comparative Production Example 1.

[0113] From this, it was found that when uwabamisou was fermented with koji mold, lactic acid bacteria, or yeast, it had a greater inhibitory effect on the formation of glycation products than non-fermented uwabamisou, and in particular, when two-stage fermentation was performed, it showed a significantly greater inhibitory effect on the formation of glycation products than non-fermented uwabamisou.

[0114] [Table 2]

[0115] <Example 3: Verification of the effect of fermentation product derived from a plant of the genus Polytrichum commune on promoting the decomposition of advanced glycation end products> (the purpose) The fermentation product derived from a plant of the genus Eleocharis of the present invention is verified to have an effect of promoting the decomposition of advanced glycation end products (AGEs).

[0116] (method) The AGEs decomposition activity was evaluated by measuring the cross-linking cleavage reaction of AGEs. Specifically, the method was performed as follows according to the method of Vasan et al. (Vasan S, et al., 1996, Nature, 382:275-278). 1-phenyl-1,2-propanedione (PPD) was used as a model substance for cross-linking in AGEs, and the Aspergillus oryzae two-stage solid / liquid fermentation extract obtained in Production Example 4, the Aspergillus oryzae non-fermentation extract obtained in Comparative Production Example 1, the Aspergillus oryzae liquid fermentation extract obtained in Comparative Production Example 5, the Aspergillus oryzae lactic acid bacteria liquid fermentation extract obtained in Comparative Production Example 8, or the Aspergillus oryzae yeast fungus liquid fermentation extract obtained in Comparative Production Example 11 were dissolved in phosphate buffer to any concentration and used as a test group for the decomposition reaction. Specifically, in each test group, the sample was mixed with PPD and incubated at 37°C for 20 hours. After incubation, 2 mol / L hydrochloric acid was added to the test group to stop the reaction, and the supernatant was collected after centrifugation. The amount of benzoic acid liberated by the decomposition of the PPD in the supernatant was measured by HPLC. The HPLC conditions were as follows: column: Inertsil ODS-3 HP (5 μm, 4.6 x 250 mm ID) (GL Sciences); eluent: water / acetonitrile / trifluoroacetic acid (volume ratio 85 / 15 / 0.05); flow rate: 0.6 mL / min; detection: UV230 nm; injection volume: 20 μL. The amount of benzoic acid originally contained in the extraction sample was also measured by HPLC under the same conditions.

[0117] The amount of benzoic acid in each test group before the reaction was subtracted from the amount of benzoic acid in the reaction solution in each test group after the reaction to calculate the molar amount (B) of benzoic acid newly generated by decomposition of PPD. Next, the molar amount (B) was divided by the molar amount (P) of PPD in the reaction solution in each test group before the reaction to obtain the value (B / P), which was defined as the AGEs cross-link cleavage rate.

[0118] (result) The results are shown in Table 3.

[0119] [Table 3]

[0120] The AGE cross-link cleavage rate in non-fermented Comparative Production Example 1 was 32.1%, whereas a higher AGE cross-link cleavage rate than non-fermented was confirmed in all of the test plots that underwent a fermentation process, namely, Production Example 4, Comparative Production Example 5, Comparative Production Example 8, and Comparative Production Example 11.

[0121] These results demonstrate that fermentation of Uwabamisō with koji mold, lactic acid bacteria, or yeast has a greater effect on promoting the decomposition of AGEs than non-fermented Uwabamisō.

[0122] <Example 4: Verification of the effect of fermented products derived from plants of the genus Eleocharis on enhancing the expression level of melatonin receptor mRNA> (the purpose) The fermented product derived from a plant of the genus Eleocharis of the present invention is verified in vitro to have an effect of enhancing the expression of the melatonin receptor gene.

[0123] (method) Human dermal fibroblasts (NB1RGB cells) were cultured until confluent in MEMα medium containing each of the Aspergillus oryzae two-stage solid / liquid fermentation extract obtained in Production Example 4, the Aspergillus oryzae non-fermented extract obtained in Comparative Production Example 1, the Aspergillus oryzae liquid fermentation extract obtained in Comparative Production Example 5, the Aspergillus oryzae lactic acid bacteria liquid fermentation extract obtained in Comparative Production Example 8, and the Aspergillus oryzae yeast fungus liquid fermentation extract obtained in Comparative Production Example 11. The control was MEMα medium not containing a sample derived from Aspergillus oryzae.

[0124] Total RNA contained in the cells after the culture was extracted using a FastGene RNA generation kit (Japan Genetics Co., Ltd.), and the amount of mRNA of the MTN1R1 (melatonin receptor) gene was measured by real-time RT-PCR. ReverTraAce qPCR RT Kit (Toyobo Co., Ltd.) was used for cDNA synthesis, and THUNDERBIRD SYBR qPCR Mix (Toyobo Co., Ltd.) was used for real-time PCR. In addition, the GAPDH gene, which is a housekeeping gene, was used as an internal standard. The primers for amplifying the MTN1R1 gene were the Fw primer shown in SEQ ID NO: 1 and the Rv primer shown in SEQ ID NO: 2. In addition, the primers for amplifying the GAPDH gene were the Fw primer shown in SEQ ID NO: 3 and the Rv primer shown in SEQ ID NO: 4. The real-time RT-PCR was performed according to a conventional method, and the MTN1R1 mRNA expression level was calculated as a ratio to the GAPDH mRNA expression level (100%). The expression rate of the MTN1R1 gene was calculated as a ratio to the mRNA expression level of the control.

[0125] (result) The results are shown in Table 4. When the non-fermented extract of Comparative Production Example 1 was added, almost no increase in the MTN1R1 mRNA expression rate was observed.

[0126] On the other hand, when the fermented Aspergillus oryzae two-stage solid / liquid fermentation extract of Production Example 4 was added at 0.005%, the MTN1R1 mRNA expression rate was increased to 194.7%, and when it was added at 0.01%, it was increased to 227.7%, more than double. When the Aspergillus oryzae liquid fermentation extract of Comparative Production Example 5 was added at 0.005%, it increased to 136.5%, and when it was added at 0.01%, it increased to 145.3%. When the Aspergillus oryzae liquid fermentation extract of Comparative Production Example 8 was added at 0.005%, it increased to 112.7%, and when it was added at 0.01%, it increased to 134.5%. When the Aspergillus oryzae liquid fermentation extract of Comparative Production Example 11 was added at 0.005%, it increased to 107.1%, and when it was added at 0.01%, it increased to 117.4%.

[0127] From the above results, it was confirmed that the fermentation of Uwabamisou with Aspergillus oryzae, Lactobacillus or yeast had an effect of enhancing the expression of melatonin receptors. This effect was confirmed to be particularly strong when Uwabamisou was fermented in two stages with bacteria.

[0128] [Table 4]

[0129] <Example 5: Verification of the decomposition effect of fermented matter derived from plants of the genus Polytrichum for advanced glycation end products in the skin> (the purpose) The fermented product derived from a plant of the genus Eleocharis of the present invention is verified in vivo to have an effect of enhancing the expression of the melatonin receptor gene.

[0130] (method) Four adult males and females with normal skin were used as subjects, and test areas of about 2 cm in diameter were set up on the inner side of the left and right forearms. The amount of AGEs in the skin of the test area was measured using an AGE Scanner (Diagnoptics Technologies BV). Before the measurement, the test area was washed, and the subjects were allowed to rest and acclimatize in the measurement room before the measurement was performed. A test lotion was prepared by adding 20% ​​of the Aspergillus oryzae two-stage solid / liquid fermentation extract of Production Example 5 to a lotion (containing carboxyvinyl polymer and paraben, adjusted to pH 6.5 with NaOH), and a control lotion solution without the fermentation extract was prepared and applied to the test area of ​​each subject twice a day, in the afternoon and at night. The test was continued for 4 weeks, and the amount of AGEs in the test area was measured after 1 week, 2 weeks, and 4 weeks. The results were calculated as a ratio with the AGEs measurement value at the start of the test taken as 100.

[0131] (result) The results are shown in Table 5. When the test lotion containing the Aspergillus oryzae two-stage solid / liquid fermentation extract of Production Example 5 was applied, a reduction in the amount of AGEs accumulated of approximately 10% was observed 4 weeks after the start of application. On the other hand, when the control lotion was applied, no clear reduction in the amount of AGEs accumulated was observed at any time after application. This demonstrated that fermentation products derived from the genus Aspergillus also exhibit an AGE-degrading effect on the skin in vivo.

[0132] [Table 5]

[0133] <Example 6: Verification of the skin elasticity improving effect of fermented matter derived from the genus Alpinia> (the purpose) The production and accumulation of AGEs leads to a decrease in skin elasticity, so we verified in vivo whether the fermented product derived from a plant of the genus Eleocharis of the present invention has an effect of improving skin elasticity.

[0134] (method) Seven adult males and females with normal skin were used as subjects, and the left and right facial cheeks were used as test areas. The elasticity of the skin in the test area was measured using Cutometer MPA580 (Courage+Khazaka), and among the measurement parameters obtained by this machine, R2 (skin elasticity) and R7 (recovery elasticity), which are the most commonly used skin elasticity parameters, were used. Measurements were performed after washing the facial cheeks and acclimating them to a test room environment (temperature 20±2 degrees, humidity 50±5%). The test lotion prepared in Example 5 was applied to one cheek, and the control lotion was applied to the other cheek. The application period was 2 weeks. The results were expressed as a ratio of the measured value at the start of the test to the measured value at 2 weeks after the start of the test, with the measured value at 100.

[0135] (result) The results are shown in Table 6. When the test lotion was applied, R2 increased by 5.7% and R7 increased by 19.6% after two weeks. On the other hand, when the control lotion was applied, R2 decreased by 0.9% and R7 increased by only 5.6%. This demonstrated that fermented products derived from the Alpinia genus have the effect of improving skin elasticity.

[0136] [Table 6]

[0137] <Example 7: Verification of the skin pigmentation improving effect of fermented products derived from the genus Alpinia> (the purpose) The production and accumulation of AGEs leads to increased skin pigmentation, so the effect of the fermented product derived from a plant of the genus Eleocharis of the present invention on improving skin pigmentation was verified using the skin of subjects.

[0138] (method) The left and right cheeks of five subjects were used as test groups, and the amount of pigmentation in the test groups was measured using API100 (Aram Huvis). A test lotion containing 20% ​​of the Aspergillus oryzae two-stage solid / liquid fermentation extract of Production Example 5 was applied to one cheek, and a control lotion without the fermentation extract was applied to the other cheek every day before going to bed. The test was performed for seven consecutive days, and the amount of pigmentation was measured using API100 on the fourth and seventh days after the test. The amount of pigmentation at the start of the test was set at 100, and the amount of pigmentation at each measurement was calculated as a ratio, which was then divided by the number of subjects to calculate the average value.

[0139] (result) The results are shown in Table 7. It was clear that the skin on the cheek to which the test lotion was applied showed an improvement in pigmentation from the fourth day after the start of the test, compared to the skin on the cheek to which the control lotion was applied.

[0140] [Table 7]

[0141] Example 8: Production of lotion containing fermented product from the genus Polytrichum and its effect (the purpose) A skin lotion containing the fermented product derived from a plant of the genus Eleocharis of the present invention is produced, and its effect on the skin is verified by a sensory test.

[0142] (method) A control lotion was prepared by mixing glycerin, 1,3-butanediol, ethanol, PEG6000, polyoxyethylene methyl glucoside (10.EO), paraoxybenzoic acid ester, dipotassium glycyrrhizinate, and polyoxyethylene hydrogenated castor oil, and a test lotion was prepared by further adding the two-stage solid / liquid fermentation extract of Aspergillus oryzae (Production Example 5) to the control lotion. Both lotions were stable without developing cloudiness for six months under conditions of 40°C and 75% RH. A sensory test was conducted on these lotions by a panel of 10 experts. Evaluation was conducted on a five-point scale for the following items.

[0143] (1) Skin moisturization: 1. Dry (1 point); 2. Slightly dry (2 points); 3. Normal (3 points); 4. Slightly moist (4 points); 5. Moist (5 points) (2) Skin smoothness: 1. Rough (1 point); 2. Slightly rough (2 points); 3. Normal (3 points); 4. Slightly smooth (4 points); 5. Smooth (5 points) (3) Sticky skin: 1. Sticky (1 point); 2. Slightly sticky (2 points); 3. Normal (3 points); 4. Slightly refreshing (4 points); 5. Refreshing (5 points)

[0144] (result) The average scores of the 10 panelists are shown in Table 8. As is clear from Table 8, the test lotion containing the Aspergillus oryzae two-stage solid / liquid fermentation extract showed superior moisturizing properties and skin smoothness to the control lotion.

[0145] [Table 8]

[0146] Example 9: Preparation of shampoo containing fermented product from the genus Polytrichum and its effect (the purpose) A shampoo containing the fermented product derived from a plant of the genus Polytrichum of the present invention is produced, and its effect on hair is verified by a sensory test.

[0147] (Methods and Results) A control shampoo was prepared by mixing PEG distearate, polysorbate 20, EDTA-2Na, TEA-cocoyl glutamate, cocamidopropyl betaine, cocamide DEA, PEG-12 dimethicone, citric acid, polysorbate 2, phenoxyethanol, and fragrance, and a test shampoo was prepared by further adding the Aspergillus oryzae two-stage solid / liquid fermentation extract of Production Example 5. A hair washing test was conducted on these shampoos by 10 expert panelists, and it was found that the test shampoo made hair feel smoother and more moisturized than the control shampoo.

Claims

1. A fermented product derived from an Elatostema plant, which is obtained by fermenting all or a part of the plant body of the Elatostema plant with bacteria or fungi.

2. The fermented product derived from a plant of the genus Eleocharis according to claim 1, wherein the fermented product is a solid fermented product obtained by fermenting a solid matter of the plant body and / or a liquid fermented product obtained by fermenting an extract of the plant body.

3. The fermented product derived from a plant of the genus Eleocharis described in claim 1 or 2, which is an inhibitor of the decomposition and production of advanced glycation end products and an enhancer of melatonin receptor gene expression.

4. A composition comprising the fermented product derived from a plant of the genus Polytrichum according to claim 1 as an active ingredient.

5. The composition according to claim 4, which is a skin elasticity improving agent, a skin pigmentation improving agent, or a cosmetic.

6. A method for producing a fermented product derived from a plant of the genus Polytrichum, comprising the steps of: a first inoculation step of inoculating all or a part of the plant body of a plant of the genus Polytrichum with bacteria or fungi; A first fermentation step in which the plant body after the first inoculation step is fermented under culture conditions for the bacteria or fungus to obtain a solid fermented product; A second inoculation step of mixing the solid fermentation product with a suspension and / or extract of all or a part of the plant body of a plant of the genus Eleocharis; and A second fermentation step in which the mixed liquid obtained after the second inoculation step is fermented under culture conditions for the bacteria or fungus to obtain a liquid fermented product. The manufacturing method comprising the steps of:

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