Skin elasticity improving agent comprising sea buckthorn fruit oil as active ingredient and use thereof

Abstract

To provide a skin elasticity improving agent.SOLUTION: A skin elasticity improving agent comprises Sea Buckthorn fruit oil, mainly derived from the pulp of Sea Buckthorn, as an active ingredient.SELECTED DRAWING: None

Description

[Technical field]

[0001] This specification relates to sea buckthorn fruit oil, which is an oil separated or extracted mainly from the pulp of sea buckthorn (also known as sea berry or Saji) (scientific name: Hippophae rhamnoides), and uses thereof. [Background technology]

[0002] The skin covers the whole body, separates the inside of the body from the outside environment, and plays an important role in maintaining homeostasis of the human body. The skin is roughly divided into three layers: the epidermis, the dermis, and the subcutaneous tissue. The epidermis is the outermost layer of the skin, and has the important function of preventing the evaporation of moisture from the body and the intrusion of foreign substances from the outside, and acting as a barrier to protect the human body from external environments such as ultraviolet rays, thereby protecting the inside of the human body. Much of the dermis is made up of extracellular matrix composed of fibrous proteins and polysaccharides. The main component of fibrous proteins is collagen, which contributes to maintaining the shape of the tissue and plays an important role in the firmness and elasticity of the skin. In addition, the collagen fibers that run throughout the skin in a mesh-like pattern are connected by elastin fibers, which are highly elastic.

[0003] Since the formation of wrinkles in the skin is related to the elasticity of the skin, which decreases with age, maintaining the elasticity of the skin is considered to be an important anti-aging factor. Collagen, the main component of fibrous proteins in the dermis, is decomposed by external factors such as ultraviolet rays and active oxygen, which causes the skin to lose elasticity, leading to a decrease in firmness and the formation of wrinkles and sagging.

[0004] Various ingredients and compositions that improve skin elasticity have also been proposed. Patent Document 1 describes that fucoidan derived from kelp improves the elasticity of skin with reduced elasticity. Patent Document 2 describes that astaxanthin improves skin elasticity. Patent Document 3 describes that tomato seed extract improves skin viscoelasticity.

[0005] On the other hand, sea buckthorn is a plant of the Elaeaceae family that grows in coastal areas, high mountains, deserts, and areas with extreme temperature differences in Europe, Mongolia, China, and Russia, and bears orange fruits. The fruits contain a lot of oil as well as juice. Therefore, after squeezing the fruits, the juice and oil (fruit oil) are often separated and used according to their purpose. The juice contains water-soluble components such as organic acids such as malic acid and ascorbic acid, amino acids such as aspartic acid, and polyphenols such as isorhamnetin. The oil contains fat-soluble components such as fatty acids, mainly palmitoleic acid, and carotenoids such as lutein, zeaxanthin, and beta-cryptoxanthin. In addition, the seeds that are removed during squeezing contain a lot of oil, but the main components are linoleic acid, alpha-linolenic acid, and oleic acid, which is a different composition from the fruit oil.

[0006] Regarding the function of sea buckthorn on the skin, it has been reported in Patent Document 4 that orally ingesting sea buckthorn juice has a moisturizing effect on the skin. In addition, palmitoleic acid, the main component of sea buckthorn oil, is known to promote collagen synthesis in cultured skin fibroblasts (Non-Patent Document 1). [Prior art documents] [Patent documents]

[0007] [Patent Document 1] JP 2003-313131 A [Patent Document 2] JP 2005-27589 A [Patent Document 3] JP 2022-149889 A [Patent Document 4] Patent No. 7131785 [Non-patent literature]

[0008] [Non-Patent Document 1] Song, IB., Gu, H., Han, HJ. et al. Omega-7 inhibits inflammation and promotes collagen synthesis through SIRT1 activation. Appl Biol Chem 61, 433-439 (2018). Summary of the Invention [Problem to be solved by the invention]

[0009] However, Patent Document 4 states that sea buckthorn juice has the effect of improving the skin barrier function and moisturizing the skin, but there is no description regarding skin elasticity, promotion of collagen synthesis gene expression, or inhibition of collagen decomposition enzyme gene expression. In addition, Non-Patent Document 1 only examines the collagen synthesis promotion effect of palmitoleic acid, and does not examine natural organic compounds other than palmitoleic acid. Furthermore, the collagen synthesis promotion effect is limited to cultured cells. In other words, there is no description regarding the relationship between sea buckthorn oil and human skin elasticity.

[0010] The present invention provides a skin elasticity improving agent containing sea buckthorn fruit oil, which is separated or extracted mainly from the flesh of sea buckthorn, as an active ingredient, and use thereof. [Means for solving the problem]

[0011] The present inventors have studied the efficacy of sea buckthorn oil in improving the elasticity of human skin when taken orally, because the efficacy of sea buckthorn oil in improving the elasticity of human skin has not yet been examined, and the degree of efficacy could not be predicted. As a result, they have found that sea buckthorn oil contributes to promoting the expression of collagen synthesis-related genes and inhibiting the expression of collagen degradation-related genes, and that oral intake of sea buckthorn oil contributes to improving the elasticity of human skin. Based on these findings, the present specification discloses the following means.

[0012] [1] A skin elasticity improving agent whose active ingredient is sea buckthorn fruit oil, which is derived mainly from the pulp of sea buckthorn. [2] The skin elasticity improving agent described in [1], which is a powder containing particles derived from the sea buckthorn fruit oil. [3] A skin elasticity improving agent as described in [1] or [2], containing palmitoleic acid in an amount of 10% by mass or more and 35% by mass or less and zeaxanthin in an amount of 0.010% by mass or more and 0.080% by mass or less relative to the total amount of the sea buckthorn fruit oil. [4] The skin elasticity improving agent according to any one of [1] to [3], which is for oral administration. [5] An expression promoter for the COL1A1 gene, which encodes collagen protein (Collagen Type I α1 chain), whose active ingredient is oil derived from sea buckthorn fruit. [6] An inhibitor of the expression of the MMP-1 gene, which encodes the collagen-degrading enzyme Matrix Metalloproteinase-1 (MMP-1), whose active ingredient is oil derived from sea buckthorn fruit. [Brief description of the drawings]

[0013] [Figure 1] FIG. 1 shows the results of an analysis of the promotion of expression of the COL1A1 gene encoding the collagen protein COL1A1 by sea buckthorn fruit oil. [Diagram 2] 1 shows the results of an analysis of the promotion of expression of the MMP-1 gene, which encodes MMP-1, a collagen decomposition enzyme, by sea buckthorn fruit oil. [Diagram 3] FIG. 1 shows the results of an analysis of the promotion of expression of the MMP-1 gene, which encodes MMP-1, a collagen decomposition enzyme, by carotenoids and palmitoleic acid contained in sea buckthorn fruit oil. [Figure 4] FIG. 1 shows the results of evaluating human skin softness following oral administration of a capsule of sea buckthorn fruit oil powder. [Diagram 5]FIG. 1 shows the results of an evaluation of the restorative power of human skin following oral administration of a capsule containing sea buckthorn fruit oil powder. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The disclosure of this specification relates to sea buckthorn fruit oil. The present inventors have found for the first time that sea buckthorn fruit oil directly contributes to promoting collagen synthesis and inhibiting collagen decomposition, and further contributes to improving skin elasticity when orally ingested. Sea buckthorn fruit oil contains palmitoleic acid as fat-soluble components, as well as carotenoids including zeaxanthin and lutein. It is believed that sea buckthorn fruit oil contributes to improving skin elasticity due to this composition.

[0015] The skin elasticity improving agent and the like will be described in detail below.

[0016] <Skin elasticity improver> The skin elasticity improving agent disclosed in the present specification contains sea buckthorn fruit oil as an active ingredient. Sea buckthorn is a plant of the Elaeaceae family, scientific name Hippophae rhamnoides, and grows in coastal areas, high mountains, deserts, and areas with extreme temperature differences in Europe, Mongolia, China, Russia, and other areas. In the present specification, sea buckthorn fruit oil is an oil made mainly from the flesh of sea buckthorn fruit. Sea buckthorn fruit oil does not use sea buckthorn seeds as a raw material, but may contain sea buckthorn peel as a raw material. Sea buckthorn fruit oil is preferably made only from the flesh of sea buckthorn fruit.

[0017] The method for obtaining sea buckthorn fruit oil is not particularly limited, and any known method can be used. For example, sea buckthorn fruit oil can be obtained by separating oil derived mainly from the pulp (hereinafter simply referred to as pulp-derived oil) from the juice (hereinafter simply referred to as fruit juice) of sea buckthorn fruit (including the skin, pulp, and seeds).

[0018] To separate the pulp-derived oil, the fruit may be squeezed to obtain juice without containing oil from the seeds, and then the peel residue and seeds may be removed to obtain a juice mainly derived from the pulp (hereinafter simply referred to as pulp juice), from which the pulp-derived oil may be separated. Note that the peel and seeds are usually easily separated from the juice by filtration, centrifugation, etc.

[0019] Alternatively, a raw material containing mainly the pulp from which at least the seeds have been removed may be obtained in advance, and the raw material may be squeezed to obtain a pulp juice, after which the pulp-derived oil may be separated from the pulp juice.

[0020] To obtain oil derived from the pulp of the fruit from the fruit juice or pulp juice, filtration, centrifugation, solvent extraction using a non-polar solvent that dissolves or disperses the sea buckthorn fruit oil such as n-hexane, supercritical carbon dioxide extraction, etc. can be used alone or in combination as appropriate. The obtained oil derived from the pulp can be used as it is as sea buckthorn fruit oil, or it can be purified by filtration or the like as necessary, or dried by vacuum drying or the like to obtain a powder.

[0021] The following is an example of a method for producing sea buckthorn fruit oil: After washing the fruit with cold water, the fruit is squeezed to extract the juice, the peel and seeds are removed as residue to obtain pulp juice, the pulp juice is centrifuged to obtain pulp-derived oil as a supernatant, and the pulp juice is filtered and then vacuum-dried to obtain sea buckthorn fruit oil as a powder.

[0022] Sea buckthorn fruit oil is commercially available as is the fruit juice and pulp used in the production of sea buckthorn fruit oil, and in some cases the fruit juice and pulp juice are also commercially available.

[0023] The sea buckthorn fruit may be dried in advance. It is also possible to obtain sea buckthorn fruit oil by appropriately obtaining oil derived from the pulp using the dried sea buckthorn fruit as a raw material.

[0024] After being obtained in the form of oil, the sea buckthorn fruit oil can be used as it is, or can be used as a paste by adding an appropriate excipient. After being obtained in the form of oil, the sea buckthorn fruit oil can be used as a solid or powder by vacuum drying, spray drying, or the like. In some cases, it is particularly effective to powderize the sea buckthorn fruit oil before use. When making the solid or powder, ingredients suitable for vacuum drying or spray drying, such as sugars, oils and fats, and antioxidants, can be added from various viewpoints.

[0025] For example, a method of powdering by spray drying can be adopted, which includes a sample liquid preparation step of preparing a sample liquid containing sea buckthorn fruit oil, sugars including at least processed starch and maltodextrin having a DE of more than 10 and not more than 20, and one or more emulsifiers selected from the group consisting of sucrose stearate having an HLB value of 15 to 16 and polyglycerol fatty acid esters having an HLB value of 11 to 16, the oil droplets having a volume-based median diameter of 100 nm to 400 nm, and a powdering step of spray-drying the sample liquid to obtain a powder. DE stands for dextrose equivalent and is an index of the saccharification rate of starch.

[0026] The sea buckthorn fruit oil can be obtained as a powder composition with improved cellular uptake and stability of oil and fat components and functional components compatible with the oil and fat components by preparing an oil-in-water emulsion of the above composition and spray-drying the emulsion as a sample liquid. The powder composition is also excellent in terms of packing properties when producing tablets and capsules, and in terms of water dispersibility. The powder composition is described in detail below.

[0027] The powder composition contains, for example, oil-derived particles containing sea buckthorn fruit oil as an oil-derived component. The powder composition may contain sugars and an emulsifier in addition to the oil-derived particles. More specifically, the powder contains sea buckthorn fruit oil as an oil-derived component, sugars containing at least processed starch and dextrin with a dextrose equivalent of more than 10 and not more than 20, and one or more emulsifiers selected from the group consisting of sucrose stearate esters with an HLB value of 15 to 16 and polyglycerin fatty acid esters with an HLB value of 11 to 16 (however, when only the polyglycerin fatty acid esters are used, the emulsifiers are contained in an amount of 6% by mass to 12% by mass based on the total amount of the oil-derived component, the sugars, and the emulsifier). The volume-based median diameter of the oil-derived component in this powder is 100 nm to 400 nm.

[0028] The median diameter of the oil droplets can be measured according to the method for measuring the average particle diameter of the oil and fat components in the powder composition already described. That is, it is measured using a laser diffraction / scattering type particle size distribution measuring device LA-960 (Horiba, Ltd.). Specifically, 1 g of oil-in-water emulsion is dissolved in 5 mL of deionized water, and 100 μl of the aqueous solution is dispersed in 20 mL of deionized water, which can be used as a sample to measure with the above device.

[0029] The sample liquid can be prepared, for example, by adding and dissolving sugars and an emulsifier in an aqueous medium heated to about 50°C or more and 70°C or less to prepare a first stock solution, adding the oil and fat components to the first stock solution cooled to a temperature at which the oil and fat components can be dispersed as oil droplets, for example, 40°C or less, or for example, 30°C or less, or for example, 25°C or more, to prepare a second stock solution, and homogenizing the second stock solution. Emulsification is possible while suppressing solidification and oxidation of the oil and fat components. A normal emulsification device can be used for homogenization to obtain an oil-in-water emulsion. In addition, the spray drying method is not particularly limited, and normal spray drying by heating, spray freeze drying, etc. can be used, but spray heat drying may be advantageous in terms of production efficiency. In addition, in the case of spray heat drying, a CL-8 spray drying device manufactured by Okawara Kakoki Co., Ltd. can be used.

[0030] The powder composition is considered to have a form of a core-shell type microcapsule (particle) having sugars and emulsifiers as a shell and an oil component as a core, or a matrix type microcapsule (particle) having an oil component as a dispersoid and sugars and emulsifiers as a matrix, and has a powder shape in a macroscopic sense. The composition of the powder composition and the median diameter of the oil component will be described below.

[0031] (oil and fat components) The oil and fat component contains edible liquid oils and fats that are liquid at least at room temperature. In this specification, room temperature refers to 15°C or higher and 25°C or lower. In terms of storage stability, it may be advantageous to contain such liquid oils and fats. Examples of liquid oils and fats include, but are not limited to, sea buckthorn fruit oil, as well as liquid oils and fats derived from plants such as linseed oil, olive oil, grape seed oil, rice oil, perilla oil, avocado oil, pumpkin seed oil, safflower oil, evening primrose oil, borage seed oil, turmeric oil, rapeseed oil, soybean oil, sesame oil, peanut oil, and rosehip oil, liquid oils and fats derived from marine organisms such as fish oil and krill oil, medium-chain triglycerides (MCT) containing medium-chain fatty acids with 6 to 12 carbon atoms as constituent fatty acids, triglycerides containing medium-chain fatty acids with 6 to 12 carbon atoms and long-chain fatty acids with 12 to 24 carbon atoms as constituent fatty acids, and mixed oils thereof. In the powder composition, one or more liquid oils can be used in combination. For example, the use of liquid oils derived from plants as the oil component may be preferable from the viewpoint of avoiding contamination of production equipment.

[0032] The oil and fat component may contain edible solid oils and fats that are solid at room temperature, such as solid oils and fats derived from plants, such as palm oil and coconut oil, as well as solid oils and fats derived from animals, such as lard and butter.

[0033] The oil and fat component may be a mixture of the liquid oil and fat and the solid oil and fat described above. In the oil and fat component, the liquid oil and fat is, for example, 50% by mass or more, for example, 60% by mass or more, for example, 65% by mass or more, for example, 70% by mass or more, for example, 75% by mass or more, for example, 80% by mass or more, for example, 85% by mass or more, for example, 90% by mass or more, for example, 95% by mass or more, for example, 97% by mass or more, for example, 98% by mass or more, for example, 99% by mass or more, for example, 100% by mass, based on the total amount of the oil and fat components. By including 50% by mass or more of the liquid oil and fat, it may be easier to obtain a suitable median size and / or it may contribute to the encapsulation state of the oil and fat components.

[0034] Furthermore, the solid fat is, for example, 30% by mass or less, for example, 25% by mass or less, for example, 20% by mass or less, for example, 15% by mass or less, for example, 10% by mass or less, for example, 5% by mass or less, for example, 1% by mass or less, relative to the total amount of the fat and oil components. If the amount of solid fat is too much, a suitable median size may not be obtained and / or the encapsulation state of the fat and oil components may be deteriorated.

[0035] The content of the oil and fat component in the powder composition is not particularly limited, and is appropriately set within a range that can obtain an oil and fat component with a predetermined median diameter.The content of the oil and fat component in the powder composition is, for example, 10% by mass or more, for example, 15% by mass or more, for example, 17% by mass or more, for example, 18% by mass or more, for example, 19% by mass or more, or for example, 20% by mass or more, based on the total amount of the oil and fat component, sugar, and emulsifier in the powder composition.The content is 30% by mass or less, for example, 25% by mass or less, for example, 23% by mass or less, for example, 22% by mass or less, or for example, 21% by mass or less. The range of the content of the oil and fat components can be appropriately selected from the above upper and lower limits, and is, for example, 15% by mass or more and 25% by mass or less, for example, 17% by mass or more and 23% by mass or less, for example, 18% by mass or more and 22% by mass or less, or for example, 19% by mass or more and 21% by mass or less.

[0036] The content of the functional ingredient in the powder composition is not particularly limited, and is appropriately set within the range of the oil and fat ingredient or as a part thereof.

[0037] (Sugars) The sugars may include at least modified starch and dextrin. The modified starch is starch modified enzymatically and / or chemically and / or physically, and is not particularly limited, but examples thereof include acetylated adipate crosslinked starch, acetylated phosphorylated crosslinked starch, acetylated oxidized starch, sodium octenyl succinate starch, starch acetate, oxidized starch, hydroxypropyl starch, hydroxypropyl phosphate crosslinked starch, phosphoric acid monoesterified phosphate crosslinked starch, phosphorylated starch, phosphate crosslinked starch, sodium starch glycolate, and sodium starch phosphate ester. The modified starch may be used alone or in combination of two or more of these, and the use of modified starch may contribute to the encapsulation and / or cellular uptake of fat and oil components. For example, the use of at least sodium octenyl succinate starch may be advantageous in that it contributes to emulsification.

[0038] Dextrin is a general term for carbohydrates that are polymerized by α-glucose through glycosidic bonds and are typically obtained, for example, by hydrolysis of starch or glycogen. As dextrin, one or more of these can be used in combination, but maltodextrins with a DE (Dextrose Equivalent, an index of the saccharification rate of starch) of more than 10 and not more than 20 may be advantageous. Maltodextrins are composed of D-glucose units of various lengths. In maltodextrin, D-glucose is linked by α1,4-glycosidic bonds, and is usually obtained as a mixture of glucose units with lengths ranging from 3 to 19.

[0039] Dextrins with a DE of 2 to 10 are sometimes simply called dextrins to distinguish them from maltodextrins. Cluster dextrins (typically highly branched dextrins made from amylopectin and the like, with high molecular weights) are also included. The DE of cluster dextrins may be about 3. Not using dextrins with a DE of 2 to 10 or cluster dextrins may be advantageous for powderization.

[0040] The sugars may include other sugars as appropriate in addition to modified starch and dextrin. Examples include monosaccharides, disaccharides, and oligosaccharides.

[0041] The content of sugar in the powder composition is not particularly limited, and is appropriately set within a range that can obtain a fat and oil component with a median diameter within a predetermined range. The content of sugar in the powder composition is, for example, 60% by mass or more, for example, 65% by mass or more, for example, 66% by mass or more, for example, 67% by mass or more, 68% by mass or more, for example, 69% by mass or more, for example, 70% by mass or more, for example, 71% by mass or more, for example, 72% by mass or more, for example, 73% by mass or more, for example, 74% by mass or more, for example, 75% by mass or more, for example, 76% by mass or more, based on the total amount of fat and oil components, sugars and emulsifiers in the powder composition. The content is, for example, 84% by mass or less, for example, 83% by mass or less, for example, 82% by mass or less, for example, 81% by mass or less, 80% by mass or less, for example, 79% by mass or less, for example, 78% by mass or less, 77% by mass or less, 76% by mass or less, for example, 75% by mass or less, and for example, 74% by mass or less. The range of the sugar content can be appropriately selected from the above upper and lower limits, and is, for example, 60% by mass or more and 84% by mass or less, for example, 70% by mass or more and 80% by mass or less, for example, 72% by mass or more and 77% by mass or less, and for example, 72% by mass or more and 75% by mass or less.

[0042] The content of modified starch and dextrin in the sugar is not particularly limited, but the total amount of modified starch and dextrin is, for example, 90% by mass or more, for example, 95% by mass or more, for example, 96% by mass or more, for example, 97% by mass or more, for example, 98% by mass or more, for example, 99% by mass or more, or for example, 100% of the total amount of sugar. The mass ratio of modified starch to dextrin in the sugar is not particularly limited, but, for example, modified starch:dextrin is 30:70 to 70:30, for example, 40:60 to 60:40, for example, 45:55 to 55:45, or for example, 50:50.

[0043] (emulsifier) As the emulsifier, at least one or more selected from the group consisting of sucrose stearate and polyglycerol fatty acid ester can be used. The emulsifier in the powder composition is appropriately selected so that the oil and fat components can be emulsified together with the sugar and encapsulated to a median size within a predetermined range.

[0044] (Sucrose stearate) Sucrose stearate, either alone or in combination with polyglycerol fatty acid ester and / or lysolecithin, may be advantageous in terms of the median size of the oil / fat component and / or cellular uptake.

[0045] From the viewpoint of improving emulsion stability, the HLB value of the sucrose stearate is, for example, 14 or more, or, for example, 15 or more. From the same viewpoint, the HLB value is, for example, 18 or less, or, for example, 17 or less, or, for example, 16 or less. Typically, the HLB value is 14 or more and 17 or less, or 15 or more and 16 or less.

[0046] The number of ester substitutions in the sucrose stearate is not particularly limited, but from the viewpoint of improving emulsion stability, the ratio of mono-substitutions (also referred to as monoesters) to the total mass of the sucrose fatty acid ester may be, for example, 70% by mass or more, for example, 75% by mass or more, for example, 80% by mass or more, for example, 85% by mass or more, for example, 90% by mass or more, for example, 95% by mass or more, or for example, 100% by mass (i.e., all sucrose fatty acid esters are monoesters). The content of monoesters in the sucrose stearate can be calculated by the following method.

[0047] (Measurement of monoester content in sucrose stearate) The ratio of the peak area at retention times of 17 to 22 minutes to the total peak area at retention times of 15 to 30 minutes in the high performance liquid chromatography shown below is calculated, and this is regarded as the content of the monoester.

[0048] <Sample preparation> Sucrose stearate is added to tetrahydrofuran in an amount of 1 mg / mL and dissolved to obtain a sample solution. <Measurement conditions> Equipment used: High performance liquid chromatography Prominence (Shimadzu Corporation) Detection method: Corona charged particle detection Column: Capcellpack C18UG120 4.6mm x 150mm (Shiseido Co., Ltd.) Column temperature: 40℃ Eluent: 0-20 min: water / methanol 50 / 50, 7 mM ammonium acetate; 20-30 min: methanol, 7 mM ammonium acetate Flow rate: 1mL / min Injection volume: 10μL

[0049] In addition, other sucrose fatty acid esters than sucrose stearate fatty acid ester can be used as appropriate. Examples of other sucrose fatty acid esters include sucrose laurate, sucrose myristic acid ester, sucrose palmitate, and sucrose oleate. As the other sucrose fatty acid esters, one or more types can be used in combination as appropriate.

[0050] As the sucrose stearate, for example, commercially available sucrose stearate can be used. Examples of the sucrose fatty acid ester include, but are not limited to, Ryoto (registered trademark) Sugar Ester S1570 (trade name, sucrose stearate, HLB value: about 15 (catalog value)), Ryoto (registered trademark) Sugar Ester S1670 (trade name, sucrose stearate, HLB value: about 16 (catalog value), DK Ester (registered trademark) F-160 (sucrose stearate, HLB channel: about 15 to 16, manufactured by Dai-ichi Kogyo Seiyaku Co., Ltd.). Other sucrose fatty acid esters that can be used include, for example, Ryoto (registered trademark) Sugar Ester P1570 (trade name, sucrose palmitate, HLB value: about 15 (catalog value), manufactured by Mitsubishi Chemical Foods Co., Ltd.), and Ryoto (registered trademark) Sugar Ester P1670 (trade name, sucrose palmitate, HLB value: about 16 (catalog value)) (all manufactured by Mitsubishi Chemical Foods Co., Ltd.). Further examples include DK Ester SS, F140, F110, F90, F70, F50, F-20W, F-10, FA-10E, Cosmelike, S-10, S-50, S-70, S-110, S-160, and S-190 (all trade names, manufactured by Dai-ichi Kogyo Seiyaku Co., Ltd.).

[0051] The content of sucrose stearate can be determined based on the total amount of the oil and fat components, sugars and emulsifiers in the powder composition. That is, the content of sucrose stearate is, for example, 0.1% by mass or more, for example, 0.5% by mass or more, for example, 0.7% by mass or more, for example, 0.8% by mass or more, for example, 1.0% by mass or more, for example, 1.2% by mass or more, for example, 1.3% by mass or more, for example, 1.4% by mass or more, or for example, 1.5% by mass or more of the total amount. The content is also 2.0 mass% or less, for example, 1.8 mass% or less, for example, 1.6 mass% or less, for example, 1.8 mass% or less, for example, 2.0 mass% or less, for example, 2.2 mass% or less, for example, 2.4 mass% or less, for example, 2.6 mass% or less, for example, 2.8 mass% or less, for example, 3.0 mass% or less, for example, 3.5 mass% or less, and for example, 4.0 mass% or less.

[0052] The range of the sucrose stearate content can be an appropriate combination of the above-mentioned lower and upper limits, and can be, for example, from 1.0% by mass to 4.0% by mass, for example, from 1.0% by mass to 3.0% by mass, for example, from 1.0% by mass to 2.0% by mass, for example, from 1.2% by mass to 1.8% by mass, or for example, from 1.4% by mass to 1.6% by mass.

[0053] In addition, when sucrose stearate is used alone as an emulsifier, or in combination with lysolecithin and / or polyglycerol fatty acid ester, the amount of use may be specified by the above lower and upper limits. For example, when used alone, a range of 1.0% by mass to 4.0% by mass may be preferable.

[0054] (Polyglycerol fatty acid ester) The polyglycerol fatty acid ester may be one or more of known polyglycerol fatty acid esters in appropriate combination. The polyglycerol fatty acid ester may be used alone or in combination with sucrose stearate and / or lysolecithin, which may be advantageous in terms of the median size, encapsulation, cellular uptake, appearance change, etc. of the oil and fat component.

[0055] From the viewpoint of further improving emulsion stability, the polyglycerol fatty acid ester has an HLB value of, for example, 10 or more, for example, 11 or more, for example, 12 or more, for example, 13 or more, for example, 14 or more, or for example, 15 or more. The HLB value is, for example, 20 or less, for example, 18 or less, or for example, 17 or less.

[0056] The HLB value of the polyglycerol fatty acid ester is preferably different from that of the sucrose fatty acid ester. By making the HLB value different, it may be advantageous for emulsion stability, encapsulation of the oil and fat component, median diameter and cellular uptake of the oil and fat component. The HLB value of the polyglyceric acid fatty acid ester may be preferably lower than the HLB value of the sucrose fatty acid ester by, for example, 1 to 4, or, for example, 1 to 3, or, for example, 1 to 2. From this viewpoint, the HLB value of the polyglycerol fatty acid ester is, for example, 11 to 13, or, for example, 12, etc.

[0057] Examples of polyglyceric acid fatty acid esters include esters of polyglycerol having an average degree of polymerization of preferably 6 to 20, more preferably 8 to 16, and even more preferably 8 to 10, and fatty acids selected from unsaturated fatty acids having 10 to 22 carbon atoms and saturated fatty acids having 8 to 22 carbon atoms. Note that the unsaturated fatty acids having 10 to 22 carbon atoms include linear unsaturated fatty acids having 10 to 22 carbon atoms and branched unsaturated fatty acids having 10 to 22 carbon atoms, and the saturated fatty acids having 8 to 22 carbon atoms include linear saturated fatty acids having 8 to 22 carbon atoms and branched saturated fatty acids having 8 to 22 carbon atoms. When the polymerization degree of the polyglycerol is within the above range, the emulsion stability is improved.

[0058] The polyglycerol fatty acid ester is preferably an ester having a fatty acid structure selected from an unsaturated fatty acid structure having 10 to 22 carbon atoms and a saturated fatty acid structure having 8 to 22 carbon atoms, and may be more preferably an ester of a fatty acid having at least one unsaturated fatty acid structure having 16 to 22 carbon atoms. Examples of fatty acids selected from unsaturated fatty acids having 10 to 22 carbon atoms include oleic acid and linoleic acid. Examples of fatty acids selected from saturated fatty acids having 8 to 22 carbon atoms include myristic acid.

[0059] The polyglycerol fatty acid ester may be a monoester or a diester, but may be preferably a monoester in some cases.

[0060] The polyglycerol fatty acid ester is not particularly limited, but may be, for example, decaglycerol laurate, decaglycerol myristic acid ester, decaglycerol palmitic acid ester, decaglycerol stearate, hexaglycerol monooleic acid ester, decaglycerol monooleic acid ester, decaglycerol linoleic acid ester, decaglycerol isostearate, hexaglycerol isostearate, decaglycerol monoisostearate, etc., and among them, it is preferable to use at least one selected from the group consisting of decaglycerol laurate, decaglycerol myristic acid ester, decaglycerol palmitic acid ester, decaglycerol stearate, polyglyceryl isostearate, and polyglyceryl oleate. Among these, decaglycerol monooleate (HLB value: 12), decaglycerol myristic acid ester (HLB value: 16), etc. may be advantageous.

[0061] As the polyglycerol fatty acid ester, commercially available products may be used, such as Poem J-0381V (manufactured by Riken Vitamin Co., Ltd.) and SY Glystar MM-750 (manufactured by Sakamoto Pharmaceutical Co., Ltd.).

[0062] The content of polyglycerol fatty acid ester can be determined based on the total amount of fat and oil components, sugars, and emulsifiers in the powder composition. For example, when used alone as an emulsifier, the content of polyglycerol fatty acid ester can be 6.0% by mass or more, for example, 7.0% by mass or more, for example, 8.0% by mass or more, or for example, 9.0% by mass or more. It can also be 12% by mass or less, for example, 11% by mass or less, or for example, 10% by mass or less. The range of polyglycerol fatty acid ester in such a case can be set by appropriately combining the above-mentioned lower and upper limits, and can be, for example, 6.0% by mass or more and 12% by mass or less, for example, 6.0% by mass or more and 11% by mass or less, for example, 7.0% by mass or more and 11% by mass or less, 8% by mass or more and 10% by mass or less, 9.0% by mass or more and 10% by mass or less, etc.

[0063] On the other hand, the content range of the polyglycerol fatty acid ester, when used in combination with sucrose stearate, can be, for example, from 1.0% by mass to 2.0% by mass, for example, from 1.2% by mass to 1.8% by mass, or for example, from 1.4% by mass to 1.6% by mass.

[0064] (Lysolecithin) The powder composition can use lysolecithin as an emulsifier in combination with sucrose stearate. Lysolecithin refers to a hydrolysate of lecithin, which is a lecithin whose main component is phospholipids, a major component of the biomembrane of animal cells, and whose alkyl chain is made single-chain by enzymatic treatment to increase its hydrophilicity. Here, lecithin is typically a mixture containing various phospholipids such as phosphatidylcholine (PC). More specifically, lysolecithin is a composition containing lysophosphatidylcholine in which one fatty acid of the phosphatidylcholine molecule has been lost by an enzyme such as phospholipase. Lysolecithin is effective in promoting emulsification of oil and fat components and adjusting the median diameter of the oil and fat components by combining it with sucrose stearate or by combining it with sucrose stearate and polyglycerol fatty acid ester. In this specification, lysolecithin includes so-called hydrogenated enzymatically decomposed lecithin, which has been subjected to hydrogenation treatment to convert the bound fatty acid into a saturated fatty acid to improve its oxidation stability.

[0065] The concentration of lysophosphatidylcholine contained in lysolecithin is, from the viewpoint of obtaining a stable oil-in-water emulsion and from the viewpoint of encapsulating the oil and fat components, for example, 18% by mass or more and 75% by mass or less, or, for example, 18% by mass or more and 65% by mass or less, or, for example, 18% by mass or more and 30% by mass or less.

[0066] The content of lysolecithin can be determined based on the total amount of the fat and oil components, sugars and emulsifiers of the powder composition. That is, the content of lysolecithin is, for example, 0.1% by mass or more, for example, 0.5% by mass or more, for example, 1% by mass or more, for example, 2% by mass or more, for example, 2.5% by mass or more, for example, 2.8% by mass or more, for example, 2.9% by mass or more of the total amount. The content is also, for example, 9% by mass or less, for example, 8% by mass or less, for example, 6% by mass or less, for example, 5% by mass or less, for example, 4.5% by mass or less, for example, 4% by mass or less, for example, 3.5% by mass or less, for example, 3.2% by mass or less, for example, 3.1% by mass or less. The range of the lysolecithin content can be an appropriate combination of the above-mentioned lower and upper limits, and can be, for example, 0.1% by mass or more and 5% by mass or less, 0.5% by mass or more and 4% by mass or less, 1% by mass or more and 4% by mass or less, or 2% by mass or more and 4% by mass or less, etc.

[0067] As the lysolecithin, for example, commercially available lysolecithin can be used, including, but not limited to, SLP-White Lyso (trade name, manufactured by Tsuji Oil Mills), SLP-White Lyso H (trade name, manufactured by Tsuji Oil Mills), Benecoat (registered trademark) BMI-40L (trade name, manufactured by Kao Corporation), and Resimal (registered trademark) EL (trade name, manufactured by Riken Vitamin Co., Ltd.).

[0068] In some cases, it may be advantageous to use only sucrose stearate as an emulsifier in the powder composition. In this case, the sucrose fatty acid ester is a sucrose fatty acid ester in which the fatty acid is stearic acid and / or the HLB value is 15 to 16, and the sucrose fatty acid ester may be, for example, 1% to 3% by mass, 1.2% to 1.7% by mass, etc., based on the total amount.

[0069] In some cases, it may be advantageous to use only polyglycerol fatty acid ester as an emulsifier for the powder composition. In this case, the polyglycerol fatty acid ester has an HLB value of 12 to 16, and the content of the polyglycerol fatty acid ester can be, for example, 5% to 10% by mass, or, for example, 6% to 10% by mass, or, for example, 7% to 10% by mass, or, for example, 8% to 10% by mass, based on the total amount.

[0070] As the emulsifier used in the powder composition, the use of sucrose stearate and lysolecithin may be advantageous in terms of the median size, encapsulation, and cellular uptake of the oil and fat component. In particular, sucrose stearate with an HLB value of 15 to 16 may be advantageous. In this case, the amount of lysolecithin may be, for example, 2% to 4% by mass, 2.5% to 3.5% by mass, and the amount of sucrose stearate may be, for example, 1% to 3% by mass, 1.2% to 1.7% by mass, relative to the total amount.

[0071] Furthermore, as an emulsifier used in the powder composition, it may be advantageous to combine sucrose stearate, lysolecithin and polyglycerin fatty acid ester. In this case, it may be advantageous for the sucrose stearate to have an HLB value of 15 to 16, and for the polyglycerin fatty acid ester to have an HLB value of 11 to 13. In this case, the sucrose stearate may be 1% by mass to 3% by mass, 1.2% by mass to 1.7% by mass, etc., and the polyglycerin fatty acid ester may be 1% by mass to 2% by mass, 1.2% by mass to 1.7% by mass, etc. The lysolecithin may be, for example, 2% by mass to 4% by mass, 2.5% by mass to 3.5% by mass, etc., relative to the total amount.

[0072] The total amount of emulsifier in the powder composition is not particularly limited, and can be, for example, 1% by mass or more and 10% by mass or less, or, for example, 1% by mass or more and 8% by mass or less, or, for example, 1.5% by mass or more and 7% by mass or less, relative to the total amount.

[0073] (Antioxidants) The powder composition may contain an antioxidant. The antioxidant is not particularly limited, and one or more known antioxidants may be appropriately used. For example, at least one compound selected from the group consisting of ascorbic acid, ascorbic acid derivatives, and salts thereof, vitamin E and its derivatives such as tocopherol, vitamin A, retinoic acid, retinol, retinol acetate, retinol palmitate, retinyl acetate, retinyl palmitate, tocopheryl retinoate, vitamin C and its derivatives, kinetin, sesamin, α-lipoic acid, coenzyme Q10, flavonoids, erythorbic acid, propyl gallate, BHT (di-n-butylhydroxytoluene), BHA (butylhydroxyanisole), tretinoin, polyphenols, SOD (superoxide dismutase), phytic acid, kouki extract, soybean extract, star anise extract, rosemary extract, etc. may be mentioned.

[0074] The content of the antioxidant is not particularly limited, but for example, from the viewpoint of encapsulation of the oil and fat components, it is from 0.1 mass % to 3 mass % relative to the total mass of the powder composition, for example, from 0.1 mass % to 2 mass %, and for example, from 0.1 mass % to 1 mass %.

[0075] (Other Ingredients) In addition to the above-mentioned components, the powder composition may contain one or more other optional components as necessary. The other components are not particularly limited as long as they can be used in, for example, pharmaceuticals, nutritional supplements, and / or food and beverages.

[0076] The powder composition may have the following characteristics. For example, the powder composition may have one or more of the following characteristics in addition to the characteristic related to the median size distribution of the oil and fat components. The median size and cellular uptake are preferable, and the powder composition may further have an encapsulation rate and / or water solubility.

[0077] (Average particle size of oil and fat components in powder composition) The average particle size of the oil and fat component contained in the powder composition can be defined as the volume-based median size. Such an average particle size is, for example, 100 nm or more and 400 nm or less. Such an average particle size may contribute to the cellular uptake of the oil and fat component. From the viewpoint of the cellular uptake of the oil and fat component and good transparency and stability of the powder composition, it may be, for example, 100 nm or more and 350 nm or less, or, for example, 100 nm or more and 300 nm or less, or, for example, 100 nm or more and 250 nm or less. In addition, in the volume-based particle size distribution of the oil and fat component, it may be preferable that 60 volume % or more of the particles are included within a range of ±50% of the volume-based median size.

[0078] The volume median diameter of the oil and fat component can be measured as the volume median diameter of oil droplets by dynamic light scattering using a laser diffraction / scattering type particle size distribution measuring device (LA-960 manufactured by Horiba, Ltd.). 1 g of the powder composition is dissolved in 5 mL of deionized water, and 100 μl of the aqueous solution is dispersed in 20 ml of deionized water, and the sample can be measured using the above device. The average particle diameter corresponds to the average particle diameter (volume median diameter) of oil droplets in an oil-in-water emulsion during the production process of the powder composition.

[0079] Sea buckthorn fruit oil contains many kinds of fatty acids. Examples include palmitoleic acid, palmitic acid, oleic acid, linoleic acid, α-linolenic acid, and stearic acid. Among them, palmitoleic acid is, for example, 10% by mass or more based on the total amount of sea buckthorn fruit oil. This is because 10% by mass or more is convenient for ensuring an effective oral intake of palmitoleic acid. For example, it is 15% by mass or more, for example, 20% by mass or more, for example, 25% by mass or more, and for example, 30% by mass or more. The upper limit is not particularly limited, but is, for example, 35% by mass or less, and for example, 30% by mass or less. The range of the content of palmitoleic acid can be set by appropriately combining the above-mentioned lower limit and upper limit, and is, for example, 10% by mass or more and 35% by mass or less, for example, 15% by mass or more and 30% by mass or less, and for example, 20% by mass or more and 30% by mass or less.

[0080] In addition to the fatty acids mentioned above, sea buckthorn fruit oil contains vitamin A, vitamin E, β-carotene, lutein, zeaxanthin, ursolic acid, oleanolic acid, etc.

[0081] Lutein contributes to improving the elasticity of the skin. The content of lutein is, for example, 0.002% by mass or more, for example, 0.003% by mass or more, for example, 0.004% by mass or more, for example, 0.005% by mass or more, or for example, 0.006% by mass or more. If the content of lutein is high, a higher improvement in skin elasticity can be expected. The upper limit is not particularly limited, but is, for example, 0.008% by mass or less, or, for example, 0.007% by mass or less. The range of the content of lutein can be set by appropriately combining the above-mentioned lower limit and upper limit, but is, for example, 0.002% by mass or more and 0.008% by mass or less, or, for example, 0.003% by mass or more and 0.007% by mass or less.

[0082] Zeaxanthin contributes to improving skin elasticity. The content of zeaxanthin in sea buckthorn fruit oil is, for example, 0.010% by mass or more, for example, 0.015% by mass or more, for example, 0.020% by mass or more, for example, 0.025% by mass or more, for example, 0.030% by mass or more, for example, 0.035% by mass or more, for example, 0.040% by mass or more, or for example, 0.050% by mass or more. If the amount of zeaxanthin is high, a higher improvement in skin elasticity can be expected. The upper limit is not particularly limited, but is, for example, 0.080% by mass or less, for example, 0.070% by mass or less, or for example, 0.060% by mass or less. The range of the zeaxanthin content can be set by appropriately combining the above-mentioned lower and upper limits, and is, for example, 0.010% by mass or more and 0.080% by mass or less, or, for example, 0.040% by mass or more and 0.070% by mass or less.

[0083] β-carotene contributes to improving skin elasticity. The content of β-carotene in sea buckthorn fruit oil is, for example, 0.010% by mass or more, for example, 0.015% by mass or more, for example, 0.020% by mass or more, and for example, 0.025% by mass or more. Higher amounts of β-carotene are expected to improve skin elasticity more. The upper limit is not particularly limited, but is, for example, 0.035% by mass or less, and for example, 0.030% by mass or less. The range of the content of β-carotene can be set by appropriately combining the above-mentioned lower limit and upper limit, but is, for example, 0.010% by mass or more and 0.030% by mass or less, or, for example, 0.015% by mass or more and 0.035% by mass or less.

[0084] Sea buckthorn fruit oil contains lutein, zeaxanthin, and beta-carotene, which contribute to improving skin elasticity, as well as palmitic acid, which also contributes to improving skin elasticity. Palmitoleic acid is a fatty acid that is a good medium for these carotenoids and is a fatty acid that is ubiquitously present in animals, including humans.

[0085] For this reason, sea buckthorn fruit oil contains multiple active ingredients for improving skin elasticity derived from carotenoids and fatty acids, and since its main component is fatty acid which contains an excellent dissolution medium and transmission medium for the active ingredients, it constitutes a useful skin elasticity improving composition in itself.

[0086] Among them, sea buckthorn fruit oil contains palmitoleic acid as one of the main fatty acids, and zeaxanthin, β-carotene, and lutein as the main carotenoids. All of these contribute to improving skin elasticity, and since they are contained as the main components, sea buckthorn fruit oil contributes greatly to improving skin elasticity. For this reason, sea buckthorn fruit oil is a useful active ingredient for skin elasticity improving agents. The skin elasticity improving agent contains carotenoids such as palmitoleic acid and zeaxanthin in the above-mentioned content or content range, and thus contributes greatly to improving skin elasticity.

[0087] Sea buckthorn fruit oil is an effective active ingredient for external use on the skin as well as for oral use. Since improvement of skin elasticity is a physical property expressed by skin tissue, even if the expression promotion of the gene encoding COL1A, a precursor protein of collagen-constituting proteins, and the expression suppression of the gene encoding the collagen-degrading enzyme MMP1 are observed in cell-level experiments, it is unclear whether skin elasticity is improved when orally administered. However, according to the present inventors, it has been found that the promotion and suppression of collagen-related gene expression at the cell level in sea buckthorn fruit oil are linked to the skin elasticity improvement effect by oral ingestion. Therefore, sea buckthorn fruit oil contributes to the improvement of skin elasticity whether it is externally applied on the skin or orally. In particular, sea buckthorn fruit oil is considered to be useful for the decrease in skin elasticity caused by ultraviolet irradiation, since it suppresses the expression of the gene encoding the collagen-degrading enzyme (MMP-1), which is enhanced by ultraviolet irradiation.

[0088] The skin elasticity improving agent is useful for improving the skin elasticity of animals such as humans. More specifically, it can improve the viscoelasticity of the skin. The viscoelasticity of the skin can be evaluated, for example, by the softness and shape recovery power of the skin. The skin elasticity improving agent improves the viscoelasticity of the skin, and further contributes to the prevention, improvement or treatment of wrinkles and sagging.

[0089] Skin softness and skin recovery can be measured by the method disclosed in the Examples below.

[0090] The skin elasticity improving agent can be used as a raw material for various foods and beverages to form foods and beverages (compositions) that can improve skin elasticity. The types of foods and beverages are not particularly limited. In addition to various ordinary foods and beverages, foods and beverages can take the form of health foods (so-called health functional foods, health supplements, nutritionally enhanced foods, nutritional supplements, etc.) and various forms of supplements (typically tablets, capsules). Capsules may be soft capsules or hard capsules.

[0091] In addition, the skin elasticity improving agent can be used as a raw material for cosmetics, medicines, and quasi-drugs to form a skin topical composition. The skin topical composition can take various known forms such as lotion, milky lotion, cream, gel, powder, paste, spray, etc. Those skilled in the art can appropriately select various components acceptable as cosmetics, medicines, and quasi-drugs according to these forms to prepare these topical agents.

[0092] The skin elasticity improving agent can be used as a raw material for a pharmaceutical intended for oral administration to constitute an oral pharmaceutical composition. Such pharmaceutical compositions can take various known forms, such as tablets, capsules, pills, powders, fine granules, granules, and liquids. Those skilled in the art can appropriately select various medicament-acceptable components according to these forms to prepare these oral pharmaceutical compositions. Oral pharmaceutical compositions are typically tablets or capsules (soft capsules and hard capsules).

[0093] The skin elasticity improving agent can also take the form of a pharmaceutical composition for local tissue administration, intradermal, subcutaneous, or intramuscular injections intended for parenteral administration. Those skilled in the art can appropriately select various medicament-acceptable components according to these forms to prepare these parenteral pharmaceutical compositions.

[0094] In these various compositions, the content of sea buckthorn fruit oil is appropriately determined. Based on the total amount of the composition, it is, for example, 1% by mass or more, for example, 10% by mass or more, for example, 15% by mass or more, or for example, 20% by mass or more. The upper limit is not particularly limited, but is, for example, 40% by mass or less, for example, 35% by mass or less, for example, 30% by mass or less, or for example, 25% by mass or less, based on the total amount of the composition. In the composition, the range of the content of sea buckthorn fruit oil can be set by appropriately combining these lower and upper limits, and can be, for example, 1% by mass or more and 40% by mass or less, for example, 10% by mass or more and 30% by mass or less, or for example, 15% by mass or more and 25% by mass or less.

[0095] The oral intake or dosage of Seabuckthorn fruit oil per day for adults can be appropriately determined according to the individual's condition, body weight, gender, age, activity of the material, intake or administration route, intake or administration schedule, dosage form, or other factors. The oral intake or dosage of Seabuckthorn fruit oil per day for adults can be, for example, 1 mg / kg body weight / day or more, and for example, 5 mg / kg body weight / day or more, and for example, 10 mg / kg body weight / day or more, and for example, 20 mg / kg body weight / day or more. The oral intake or dosage of Seabuckthorn fruit oil per day for adults can be, for example, 1000 mg / kg body weight / day or less, and for example, 500 mg / kg body weight / day or less, and for example, 250 mg / kg body weight / day or less, and for example, 100 mg / kg body weight / day or less. Also, the range of the oral intake or dosage of Seabuckthorn fruit oil per day for adults can be, for example, 1 to 1000 mg / kg body weight / day, and for example, 5 to 500 mg / kg body weight / day, and for example, 10 to 250 mg / kg body weight / day, and for example, 20 to 100 mg / kg body weight / day.

[0096] <COL1A1 gene expression promoter> The COL1A1 gene expression promoter disclosed herein contains Seabuckthorn fruit oil as an active ingredient. Here, the COL1A1 gene encodes the α1 chain (Collagen Type I α1) of type I collagen having a triple helix composed of two α1 chains and one α2 chain. Promoting the expression of the COL1A1 gene means promoting the synthesis of collagen that contributes to the elasticity of the skin. Here, the amino acid sequence of human COL1A1 encoded by the human COL1A1 gene and the nucleotide sequence of mRNA are accessible at the NIH site (https: / / www.ncbi.nlm.nih.gov) accession numbers NP_000079 and NM_000088, respectively (https: / / www.ncbi.nlm.nih.gov / protein / NP_000079.2) (https: / / www.ncbi.nlm.nih.gov / nuccore / NM_000088.4).

[0097] The gene expression promoter, like the skin elasticity improver, contains various forms of Sebaceous zone fruit oil as an active ingredient and can take various forms such as oil, paste, powder, etc.

[0098] <MMP-1 Gene Expression Inhibitor> The MMP-1 gene expression inhibitor disclosed in this specification contains Sebaceous zone fruit oil as an active ingredient. Here, the MMP-1 gene encodes Matrix Metalloproteinase-1, MMP-1, an enzyme involved in the degradation of collagen. The inhibitor of MMP-1 gene expression means the inhibition of the degradation of collagen that contributes to the elasticity of the skin. Here, the amino acid sequence of human MMP-1 encoded by the human MMP-1 gene and the nucleotide sequence of mRNA are accessible at the NIH site (https: / / www.ncbi.nlm.nih.gov) accession numbers NP_002412.1 and NM_002421.4 (https: / / www.ncbi.nlm.nih.gov / protein / NP_002412.1) and https: / / www.ncbi.nlm.nih.gov / nuccore / NM_002421.4), respectively.

[0099] The gene expression inhibitor, like the skin elasticity improver, contains various forms of Sebaceous zone fruit oil as an active ingredient and can take various forms such as oil, paste, powder, etc.

[0100] The above gene expression promoter and gene expression inhibitor, like the skin elasticity improver, can be used as raw materials for various compositions to constitute various compositions. These compositions can be manufactured, used, and administered in the same manner as the skin elasticity improver.

Example

[0101] Hereinafter, examples will be described as specific examples to more specifically explain the disclosure of this specification. The following examples are for explaining the disclosure of this specification and do not limit its scope.

Example

[0102] <Production of Sea Buckthorn Fruit Oil> Sea buckthorn fruit was washed with cold water, then squeezed, and the skin and seeds were removed. The juice was centrifuged to separate the oil and juice. The oil was filtered through a filter and then vacuum dried to obtain sea buckthorn fruit oil. The palmitoleic acid content of the obtained sea buckthorn fruit oil was 29.1% by mass, the lutein content was 0.0045% by mass, and the zeaxanthin content was 0.063% by mass. EXAMPLES

[0103] <Sea buckthorn fruit oil promotes collagen (COL1A1) gene expression> Normal human dermal fibroblasts were cultured in 5% FBS-containing DMEM medium at 5% CO2 and 37℃ until confluent. The medium was replaced, sea buckthorn fruit oil was added at 1 ppm, and the cells were cultured for 24 hours. RNA was extracted from the samples and collagen (COL1A1) mRNA expression was analyzed by real-time RT-PCR. Dimethyl sulfoxide (DMSO) was used instead of sea buckthorn fruit oil as a control. The results are shown in Figure 1.

[0104] As shown in Figure 1, the amount of mRNA encoding COL1A1 increased when sea buckthorn fruit oil was added to the medium, indicating that sea buckthorn fruit oil promotes the expression of genes related to collagen synthesis. EXAMPLES

[0105] <Inhibitory effect of sea buckthorn fruit oil on collagen decomposition enzyme (MMP-1) gene expression by UV irradiation> Normal human skin fibroblasts were cultured in 5% FBS-containing DMEM medium at 5% CO2 and 37°C until confluent. The medium was replaced and sea buckthorn fruit oil was added to a concentration of 1 ppm. After 24 hours of culture, the cells were washed with HBSS buffer and incubated at 100 mJ / cm 2 After UV-B irradiation, the cells were cultured for 24 hours in DMEM medium containing 0.5% FBS, and RNA was extracted from the cells, and gene expression of collagen degrading enzyme (MMP-1) was analyzed by real-time RT-PCR. As a control, dimethyl sulfoxide (DMSO) was used instead of sea buckthorn fruit oil. The results are shown in Figure 2.

[0106] As shown in Figure 2, it was found that the expression of the collagen-degrading enzyme MMP-1 after UV irradiation could be suppressed by adding sea buckthorn fruit oil to the culture medium. EXAMPLES

[0107] <Inhibitory effect of carotenoids and palmitoleic acid on collagen decomposition enzyme (MMP-1) gene expression by UV irradiation> Normal human skin fibroblasts were cultured in 5% FBS-containing DMEM medium at 5% CO2 and 37°C until confluent. The medium was replaced and sea buckthorn fruit oil was added so that the main carotenoids (zeaxanthin, β-carotene, and lutein) contained in 1 ppm of sea buckthorn fruit oil were each added at 0.63 ppb, and palmitoleic acid was added at 0.291 ppm. After 24 hours of culture, the cells were washed with HBSS buffer and incubated at 100 mJ / cm 2The cells were irradiated with 1000 ppb of UV-B. 0.63 ppb corresponds to the zeaxanthin content in the sea buckthorn fruit oil used in Example 2. 0.291 ppb corresponds to the palmitoleic acid content in the sea buckthorn fruit oil. After UV-B irradiation, RNA was extracted from the cells cultured for 24 hours in DMEM medium containing 0.5% FBS, and gene expression of collagen decomposition enzyme (MMP-1) was analyzed by real-time RT-PCR. As a control, dimethyl sulfoxide (DMSO) was used instead of sea buckthorn fruit oil. The results are shown in Figure 3. The letters in the graph indicate significant differences when different. (P<0.05, n=3)

[0108] As shown in Figure 3, β-carotene, lutein, zeaxanthin, and palmitoleic acid each suppressed MMP-1 gene expression against UV irradiation. These results indicate that sea buckthorn fruit oil is useful for suppressing MMP-1 gene expression. EXAMPLES

[0109] <Sea buckthorn fruit oil improves skin elasticity> [1] Production of test foods and comparison foods Sea buckthorn fruit oil was suspended in water, starch hydrolysate, modified starch, sucrose fatty acid ester, glycerin fatty acid ester, antioxidant, and enzymatically hydrolyzed lecithin, so that the sea buckthorn fruit oil was 20% by mass of the total solids.

[0110] Specifically, 150 kg of water was heated to 60°C, and modified starch (CAPSUL, Ingredion, 11.0 kg), starch hydrolysate (TK-16, Matsutani Chemical Industry, 11.0 kg), sucrose fatty acid ester (HLB value 16, sucrose stearate ester) (DK Ester F-160, Daiichi Kogyo Seiyaku, 0.45 kg), glycerin fatty acid ester (HLB value 12) (Poem J-0318V, Riken Vitamin, 0.45 kg), and enzymatically hydrolyzed lecithin (SLP-White Lyso, Tsuji Oil Mill, 0.90 kg) were added to the water and dissolved. After confirming that the emulsifiers and other ingredients had dissolved, the temperature of the liquid was lowered to 40°C, and then 6.0 kg of sea buckthorn fruit oil or soybean oil as oil components and 0.09 kg of an antioxidant (Aircoat CE, manufactured by Mitsubishi Chemical Corporation) were added to the solution of the emulsifiers and other ingredients.

[0111] The liquid was then treated for 60 minutes at 8,000 rpm using a homogenizer (TK Homomixer, manufactured by Primix Corporation) in a nitrogen-filled chamber, and the treated liquid was then further treated three times using a high-pressure homogenizer (Nano-Vita C-ES, manufactured by Yoshida Kikai Kogyo Co., Ltd.) at a treatment pressure of 100 MPa to obtain an oil-in-water emulsion.

[0112] The volume-based median diameter of the oil and fat components in the oil-in-water emulsion was measured using a laser diffraction / scattering particle size distribution analyzer “LA-960” (manufactured by Horiba, Ltd.) by dissolving 1 g of the oil-in-water emulsion in 5 mL of deionized water, diluting 100 μL of this solution with 20 mL of deionized water, and analyzing the diluted solution as a sample, which was 140 nm.

[0113] This oil-in-water emulsion was spray-dried using a spray dryer (CL-8 manufactured by Okawara Kakoki Co., Ltd.) at a tower temperature of 140°C and an atomizer rotation speed of 20 Hz to produce sea buckthorn fruit oil powder or soybean oil powder.

[0114] The obtained sea buckthorn fruit oil powder and soybean oil powder were filled into hard capsules in the proportions shown in Table 1 to produce test foods 1 and 2. Test food 1 contained 9 mg of sea buckthorn fruit oil per capsule, and test food 2 contained 27 mg of sea buckthorn fruit oil per capsule. The comparative food contained 27 mg of soybean oil per capsule.

[0115] [Table 1]

[0116] [2] A double-blind study on improving skin elasticity with capsules containing sea buckthorn fruit oil powder A double-blind, parallel-group comparative study was conducted to verify that oral intake of test foods 1, 2, and the comparison food for four weeks improves skin elasticity.

[0117] (1) Study design The randomized, placebo-controlled, double-blind, parallel-group comparative study was conducted over a four-week period from late January to early March 2023. This period is seasonally dry, and skin elasticity and recovery ability are typically reduced during this period (see comparative food intervention group).

[0118] (2) Allocation The 99 subjects were randomly assigned to three groups, with 33 subjects in each of the test food 1 intervention group, test food 2 intervention group, and comparison food intervention group. Table 2 shows the background of the subjects by group. No significant differences were observed in age or BMI among the three groups.

[0119] [Table 2]

[0120] (3) Intervention, intervention period The subjects were prevented from distinguishing between the three groups based on appearance, taste, or smell, and were asked to take four hard capsules per day with water at any time of their choice. Intervention period: 4 weeks

[0121] (4) Evaluation items and measurements The evaluation item was skin viscoelasticity, which was measured before and 4 weeks after intake using Cutometer dual (registered trademark) MPA 580 (Courage + Khazaka electronic GmbH, Germany). The measurement was performed by placing the measurement probe of the Cutometer on the left cheek of the subject, sucking the skin with negative pressure from the opening of the probe for a certain period of time, and then releasing it. The maximum change value during negative pressure suction was R0 (unit: mm), and the return value after releasing the negative pressure was R8 (unit: mm).

[0122] R0 indicates the hardness (softness) of the skin, and R8 indicates the ability of the skin to return to its original state; the higher the value of either index, the better the condition.

[0123] (5) Statistical methods All statistical analyses were two-sided, with a significance level set at 5%. SPSS Statistics version 23 or higher was used, with other validated statistical software used as necessary. All pairwise comparisons were with the comparison food intervention group.

[0124] (6) Results (6-1) Skin viscoelasticity measurement The mean, SD, Med, Min, Max, intergroup difference, SE, 95%CI-, 95%CI+, and statistical analysis results of skin viscoelasticity measurements are shown in Figures 4 and 5. Figure 4 shows the analysis results of flexibility (R0), and Figure 5 shows the analysis results of recovery force (R8).

[0125] As shown in Figure 4, regarding skin flexibility, after 4 weeks of intake, the initial maximum amplitude (R0) of the left cheek was significantly higher in the test food 2 intervention group compared to the comparison food intervention group (comparison food intervention group: 0.1726 ± 0.0473 mm, test food 1 intervention group: 0.2319 ± 0.1631 mm, difference between groups: 0.0560 mm [0.0114, 0.1006], P = 0.014). A tendency to higher values ​​was also confirmed in the test food 1 intervention group (P = 0.074).

[0126] Furthermore, as shown in Figure 5, with regard to the skin's recovery ability, after 4 weeks of intake, the total recovery (R8) of the first curve on the left cheek was significantly higher in the test food 1 intervention group (test food 1 intervention group: 0.1548 ± 0.1095 mm, difference between groups: 0.0398 mm [95% 0.0006, 0.0791], P = 0.047) and the test food 2 intervention group (comparison food intervention group: 0.1160 ± 0.0372 mm, test food 2 intervention group: 0.1704 ± 0.1514 mm, difference between groups: 0.0533 mm [0.0141, 0.0926], P = 0.008)) compared to the comparison food intervention group.

[0127] In addition, the average values ​​of R0 and R8 in the comparative food intervention group from before the start of intake to 4 weeks after intake were 0.2348 mm and 0.1726 mm for R0, and 0.1798 mm and 0.1160 mm for R8, respectively, and both indices decreased during the test period. From this, it was considered that in this study, the test food 1 intervention group and the test food 2 intervention group suppressed the decrease in the skin's flexibility and shape recovery ability during the test period. [Industrial Applicability]

[0128] The skin elasticity improving agents and the like disclosed in the present specification can be incorporated into foods, cosmetics, and pharmaceuticals and are industrially useful.

Claims

1. A skin elasticity improving agent whose active ingredient is sea buckthorn fruit oil, which is mainly derived from the flesh of sea buckthorn.

2. The skin elasticity improving agent according to claim 1, which is a powder containing particles derived from the sea buckthorn fruit oil.

3. 2. The skin elasticity improving agent according to claim 1, which contains palmitoleic acid in an amount of 10% by mass or more and 35% by mass or less and zeaxanthin in an amount of 0.010% by mass or more and 0.080% by mass or less relative to the total amount of the sea buckthorn fruit oil.

4. The skin elasticity improving agent according to claim 1, which is for oral administration.

5. An expression promoter for the COL1A1 gene that codes for collagen protein (Collagen Type I α1 chain), comprising oil derived from sea buckthorn fruit as an active ingredient.

6. An expression inhibitor of the MMP-1 gene, which encodes the collagen-degrading enzyme Matrix Metalloproteinase-1 (MMP-1), containing oil derived from sea buckthorn fruit as an active ingredient.

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