Skin elasticity improving agent containing sea buckthorn fruit oil as an active ingredient and its use

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 mainly separated or extracted from the pulp of sea buckthorn (also known as sea berry or saji, scientific name: Hippophae rhamnoides), and its uses.

Background Art

[0002] The skin covers the whole body, separates the internal and external environments, and plays an important role in maintaining the constancy 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, preventing the evaporation of body moisture and the invasion of foreign substances from the outside, serving as a barrier to protect the human body from external environments such as ultraviolet rays, and having a necessary function to protect the interior of the human body. Most of the dermis is occupied by an extracellular matrix composed of fibrous proteins, polysaccharides, etc. The main component of the fibrous protein is collagen, which contributes to maintaining the shape of the tissue and plays an important role in the firmness and elasticity of the skin. Also, the collagen fibers stretched in a network are joined together by elastic elastin fibers.

[0003] Since the elasticity of the skin, which decreases with aging, is involved in the formation of skin wrinkles, maintaining the elasticity of the skin is considered important as anti-aging. Collagen, which is the main component of the fibrous proteins in the dermis, is promoted to decompose by external factors such as ultraviolet rays and reactive oxygen species, whereby the skin loses its elasticity and is involved in the decrease of firmness, the formation of wrinkles and sagging.

[0004] Various components and compositions for improving the elasticity of the skin have also been proposed. Patent Document 1 describes that fucoidan derived from kelp improves the decreased elasticity of the skin. Patent Document 2 describes that astaxanthin improves the elasticity of the skin. Patent Document 3 describes that a tomato seed extract improves the viscoelasticity of the skin.

[0005] On the other hand, sea buckthorn is a plant belonging to the Elaeagnaceae family that grows in coastal areas, high mountains, deserts, and regions with extreme temperature fluctuations, including Europe, Mongolia, China, and Russia, and bears orange fruits. The fruit contains not only juice but also a large amount of oil. Therefore, after pressing the fruit, the juice and oil (fruit oil) are often separated and used separately depending on the 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 fatty acids mainly consisting of palmitoleic acid, and fat-soluble components such as carotenoids such as lutein, zeaxanthin, and β-cryptoxanthin. In addition, the seeds that are removed during pressing contain a large amount of oil, but the main components are linoleic acid, α-linolenic acid, and oleic acid, and the composition of the seeds is different from that of the fruit oil.

[0006] Regarding the skin's functions, Patent Document 4 reports that oral intake of sea buckthorn juice has a moisturizing effect on the skin. Furthermore, 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] Japanese Patent Publication No. 2003-313131 [Patent Document 2] Japanese Patent Publication No. 2005-27589 [Patent Document 3] Japanese Patent Publication No. 2022-149889 [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). [Overview of the project] [Problems that the invention aims to solve]

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

[0010] This specification provides a skin elasticity improving agent and its uses, which contains sea buckthorn fruit oil, mainly separated or extracted from the pulp of sea buckthorn, as an active ingredient. [Means for solving the problem]

[0011] The inventors of this invention investigated the efficacy of sea buckthorn oil on improving skin elasticity in humans, as its efficacy in improving skin elasticity had not yet been studied, and the degree of its efficacy could not be predicted. As a result, they found that sea buckthorn oil contributes to promoting the expression of collagen synthesis-related genes and suppressing the expression of collagen degradation-related genes, and that oral intake contributes to improving the elasticity of human skin. Based on these findings, this specification discloses the following methods.

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

[0013] [Figure 1] This figure shows the results of an analysis of the expression enhancement of the COL1A1 gene, which encodes the collagen protein COL1A1, by sea buckthorn fruit oil. [Figure 2] This figure shows the results of an analysis of the expression enhancement of the MMP-1 gene, which encodes the collagen-degrading enzyme MMP-1, by sea buckthorn fruit oil. [Figure 3] This figure shows the results of an analysis of the increased expression of the MMP-1 gene, which encodes the collagen-degrading enzyme MMP-1, by carotenoids and palmitoleic acid contained in sea buckthorn fruit oil. [Figure 4] This figure shows the results of evaluating the flexibility of human skin after oral administration of capsules containing sea buckthorn fruit oil powder. [Figure 5]It is a diagram showing the evaluation results of the restorative power of human skin by oral administration of a capsule preparation of sea buckthorn fruit oil powder.

Mode for Carrying Out the Invention

[0014] The disclosure of this specification relates to sea buckthorn fruit oil. According to the present inventors, it has been found for the first time that sea buckthorn fruit oil directly contributes to promoting the synthesis of collagen and suppressing the degradation of collagen, and further contributes to improving the elasticity of the skin even when orally ingested. Sea buckthorn fruit oil contains, as fat-soluble components, in addition to palmitoleic acid, carotenoids including zeaxanthin and lutein. Sea buckthorn fruit oil is considered to contribute to improving the elasticity of the skin due to such a composition.

[0015] Hereinafter, the skin elasticity improver and the like will be described in detail.

[0016] <Skin Elasticity Improver> The skin elasticity improver disclosed in this specification contains sea buckthorn fruit oil as an active ingredient. Sea buckthorn is a plant of the Elaeagnaceae family with the scientific name Hippophae rhamnoides, and it inhabits coastal areas, mountains, deserts, and regions with drastic temperature differences such as Europe, Mongolia, China, and Russia. In this specification, sea buckthorn fruit oil is an oil mainly made from the pulp of sea buckthorn fruit. Sea buckthorn fruit oil does not use the seeds of sea buckthorn as a raw material, but may contain the fruit skin of sea buckthorn as a raw material. Sea buckthorn fruit oil preferably uses only the pulp of sea buckthorn fruit as a raw material.

[0017] To obtain sea buckthorn fruit oil, known methods can be employed, but are not particularly limited. For example, sea buckthorn fruit oil can be obtained by separating the oil mainly derived from the fruit pulp (hereinafter simply referred to as "fruit pulp oil") from the juice (hereinafter simply referred to as "fruit juice") of sea buckthorn fruit (including the peel, pulp, and seeds).

[0018] To separate the oil derived from the fruit pulp, the fruit may be pressed to obtain juice, ensuring that the oil from the seeds is not included. After that, the peel residue and seeds may be removed to obtain juice mainly derived from the fruit pulp (hereinafter simply referred to as "pulp juice"), and the oil derived from the fruit pulp may be separated from this pulp juice. The peel and seeds can usually be easily separated from the juice by filtration or centrifugation.

[0019] Alternatively, a raw material mainly consisting of fruit pulp with at least the seeds removed may be obtained in advance, and after obtaining fruit pulp juice by pressing or other means, the oil derived from the fruit pulp may be separated from this fruit pulp juice.

[0020] To obtain oil derived from the pulp of fruit juice or pulp juice, filtration, centrifugation, solvent extraction using a nonpolar solvent that dissolves or disperses sea buckthorn fruit oil, such as n-hexane, and supercritical carbon dioxide extraction can be used individually or in appropriate combinations. The obtained pulp-derived oil can be used as is as sea buckthorn fruit oil, or it may be purified by filtration or other methods as needed, or dried by vacuum drying or other methods to obtain a powder.

[0021] One example of a method for producing sea buckthorn fruit oil is as follows: The fruit is washed with cold water, pressed to extract the juice, the peel and seeds are removed as residue to obtain the pulp and juice, the pulp and juice is centrifuged to obtain the pulp-derived oil as the supernatant, and then filtered and vacuum-dried to obtain sea buckthorn fruit oil in powder form.

[0022] Sea buckthorn fruit oil is commercially available on its own, and the fruit juices and pulp juices used in the production process of sea buckthorn fruit oil are also sometimes commercially available.

[0023] The sea buckthorn fruit may be dried beforehand. It is also possible to obtain sea buckthorn fruit oil by using dried sea buckthorn fruit as a raw material and obtaining the appropriate amount of oil derived from the fruit pulp.

[0024] Sea buckthorn fruit oil can be used as is after obtaining it in oil form, or it can be used as a paste by adding appropriate excipients. Furthermore, after obtaining sea buckthorn fruit oil as oil, it can be used as a solid or powder by vacuum drying, spray drying, etc. In some cases, using sea buckthorn fruit oil in powder form is particularly effective. When solidifying or powdering, components such as sugars, oils, and antioxidants suitable for vacuum drying or spray drying can be included from various perspectives.

[0025] For example, as a method for powdering by spray drying, a method can be employed that comprises a sample solution preparation step of preparing a sample solution containing sea buckthorn fruit oil, sugars containing at least modified starch and maltodextrin with an HLB value of more than 10 and 20 or less, and one or more emulsifiers selected from the group consisting of sucrose stearate ester with an HLB value of 15 or more and polyglycerin fatty acid ester with an HLB value of 11 or more and 16 or less, wherein the median diameter of the oil droplets based on volume is 100 nm or more and 400 nm or less, and a powdering step of spray drying the sample solution to obtain a powder. DE stands for Dextrose Equivalent, and is an indicator of the saccharification rate of starch.

[0026] Sea buckthorn fruit oil can be obtained as a powder composition with improved cellular uptake and stability of oily components and functional components compatible with oily components by preparing an oil-in-water emulsion of this composition and spray-drying it as a sample solution. Furthermore, it exhibits excellent filling properties when manufacturing tablets and capsules, as well as superior water dispersibility. The powder composition is described in detail below.

[0027] The powder composition contains, for example, oil-derived particles including sea buckthorn fruit oil, which is an oil component. In addition to the oil-derived particles, the powder composition may also contain sugars and emulsifiers. More specifically, it contains a powder containing sea buckthorn fruit oil, which is an oil component; sugars containing at least modified starch and dextrin with a dextrose equivalent of more than 10 and 20 or less; and one or more emulsifiers selected from the group consisting of sucrose stearate ester with an HLB value of 15 to 16 and polyglycerin fatty acid ester with an HLB value of 11 to 16 (however, if only the polyglycerin fatty acid ester is used, it shall be contained in an amount of 6% to 12% by mass relative to the total amount of the oil component, sugars and emulsifiers). Furthermore, the volume-based median diameter of the oil component in this powder is 100 nm to 400 nm.

[0028] The median diameter of the oil droplet can be measured in accordance with the method for measuring the average particle size of the oil components in the powder composition, as previously described. Specifically, it is measured using a laser diffraction / scattering particle size distribution analyzer LA-960 (Horiba, Ltd.). For the droplet, 1 g of oil-in-water emulsion is dissolved in 5 mL of deionized water, and 100 μl of this aqueous solution is dispersed in 20 mL of deionized water. This sample can then be measured using the above-mentioned apparatus.

[0029] The sample solution can be prepared, for example, by adding sugars and an emulsifier to an aqueous medium heated to approximately 50°C to 70°C to dissolve them and prepare a first stock solution; then, after the first stock solution has been cooled to a temperature at which the oil components can disperse as oil droplets, for example, 40°C or below, or for example, 30°C or below, or for example, 25°C or above, the oil components are added to prepare a second stock solution; and finally, the second stock solution is homogenized. This suppresses the solidification and oxidation of the oil components, enabling emulsification. For homogenization to obtain an oil-in-water emulsion, a conventional emulsification apparatus can be used. Furthermore, while there are no particular limitations on the spray drying method, conventional spray drying by heating, spray freeze-drying, etc., can be used, but spray heating drying may be advantageous from the viewpoint of production efficiency. In the case of spray heating drying, a spray drying apparatus such as the CL-8 manufactured by Okawara Chemical Machinery Co., Ltd. can be used.

[0030] The powder composition is thought to have the form of microcapsules (particles) of either a core-shell type, where sugars and emulsifiers form the shell and oil components form the core, or a matrix type, where oil components form the dispersed phase and sugars and emulsifiers form the matrix, and macroscopically it has the form of a powder. The composition of the powder composition and the median diameter of the oil components will be explained below.

[0031] (oil and fat components) The oil and fat components include at least edible liquid oils that are liquid at room temperature. In this specification, room temperature refers to a temperature between 15°C and 25°C. From the viewpoint of storage stability, it may be advantageous to include such liquid oils. Examples of liquid oils are not particularly limited, but include sea buckthorn fruit oil, as well as plant-derived liquid oils such as linseed oil, olive oil, grapeseed 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; marine-derived liquid oils such as fish oil and krill oil; medium-chain triglycerides (MCTs) whose constituent fatty acids are medium-chain fatty acids with 6 to 12 carbon atoms; triglycerides whose constituent fatty acids are medium-chain fatty acids with 6 to 12 carbon atoms and long-chain fatty acids with 12 to 24 carbon atoms; and mixed oils thereof. In powder compositions, one or more liquid oils and fats can be used in combination. As for the oil and fat component, using, for example, plant-derived liquid oils and fats may be preferable from the viewpoint of avoiding contamination of manufacturing equipment.

[0032] The oil and fat components may include edible solid fats that are solid at room temperature. Examples of such fats and fats include plant-derived solid fats such as palm oil and coconut oil, as well as animal-derived solid fats such as lard and butter.

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

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

[0035] The content of oil and fat components in the powder composition is not particularly limited and is set appropriately within a range in which oil and fat components with a median diameter within a predetermined range can be obtained. The content of oil and fat components 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, for example, 20% by mass or more, based on the total amount of oil and fat components, sugars and emulsifiers in the powder composition. Furthermore, 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, for example, 21% by mass or less. Furthermore, the range of oil and fat content can be appropriately selected from the above upper and lower limits, 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, and for example, 19% by mass or more and 21% by mass or less.

[0036] The content of functional components in the powder composition is not particularly limited and can be appropriately set within or as part of the range of oil and fat components.

[0037] (Sugars) The sugars may include at least modified starch and dextrin. Modified starch is starch that has been enzymatically and / or chemically and / or physically modified, and is not particularly limited, but examples include acetylated adipic acid cross-linked starch, acetylated phosphorylated cross-linked starch, acetylated oxidized starch, sodium octenyl succinate starch, starch acetate, oxidized starch, hydroxypropyl starch, hydroxypropyl phosphate cross-linked starch, phosphate monoesterified phosphate cross-linked starch, phosphorylated starch, phosphate cross-linked starch, sodium starch glycolate, and sodium starch phosphate ester. One or more of these can be used as modified starch, and the use of modified starch may contribute to the encapsulation and / or cell uptake of lipid components. For example, using at least sodium octenyl succinate starch may be advantageous as it can contribute to emulsification.

[0038] Dextrins are carbohydrates obtained by polymerizing α-glucose via glycosidic bonds, typically from the hydrolysis of starch or glycogen. While dextrins can be used individually or in combination of two or more types, maltodextrins with a DE (Dextrose Equivalent, an indicator of starch saccharification rate) of more than 10 and less than or equal to 20 may be advantageous. Maltodextrins are composed of D-glucose units of varying lengths. In maltodextrins, D-glucose is linked by α1,4-glycosidic bonds, and they are usually obtained as a mixture of glucose units ranging in length from 3 to 19.

[0039] Furthermore, 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, etc., with high molecular weights) are another example. The DE of cluster dextrins can be around 3. Avoiding dextrins with a DE of 2 to 10 or cluster dextrins may be advantageous for powder formation.

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

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

[0042] The content of modified starch and dextrin in the sugars is not particularly limited, but the total amount of modified starch and dextrin is, for example, 90% or more by mass, 95% or more by mass, 96% or more by mass, 97% or more by mass, 98% or more by mass, 99% or more by mass, or 100% by mass of the total amount of sugars. Furthermore, the mass ratio of modified starch to dextrin in the sugars is not particularly limited, but for example, the modified starch:dextrin ratio 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 an emulsifier, at least one or more selected from the group consisting of sucrose stearate esters and polyglycerol fatty acid esters can be used. The emulsifier in the powder composition is appropriately selected together with sugars to emulsify the oil and fat components and encapsulate the oil and fat components within a predetermined median diameter range.

[0044] (Sucrose stearate ester) Sucrose stearate may be advantageous, either alone or in combination with polyglycerol fatty acid esters and / or lysolecithin, in terms of median diameter and / or cellular uptake of lipid components.

[0045] From the viewpoint of improving emulsification stability, the HLB value of sucrose stearate should be, for example, 14 or higher, or 15 or higher. Similarly, the HLB value should be, for example, 18 or lower, 17 or lower, or 16 or lower. Typically, the HLB value is between 14 and 17, or between 15 and 16.

[0046] The number of ester substitutions in sucrose stearate is not particularly limited, but from the viewpoint of improving emulsification stability, the proportion of monosubstituted products (also called monoesters) to the total mass of sucrose fatty acid stearate 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 monoester content in sucrose stearate can be calculated by the following method.

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

[0048] <Sample Preparation> Sucrose stearate is added to tetrahydrofuran in an amount equal to 1 mg / mL and dissolved to obtain the 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] Other sucrose fatty acid esters besides sucrose stearate fatty acid ester can also be used as appropriate. Examples of other sucrose fatty acid esters include sucrose laurate ester, sucrose myristic acid ester, sucrose palmitate ester, and sucrose oleic acid ester. One or more of these other sucrose fatty acid esters can be used in appropriate combinations.

[0050] As the sucrose stearate ester, for example, commercially available sucrose stearate esters can be used. While not particularly limited, examples include Ryoto® Sugar Ester S1570 (product name, sucrose stearate ester, HLB value: approximately 15 (catalog value)), Ryoto® Sugar Ester S1670 (product name, sucrose stearate ester, HLB value: approximately 16 (catalog value)), and DK Ester® F-160 (sucrose stearate ester, HLB channel: approximately 15-16, manufactured by Daiichi Kogyo Seiyaku Co., Ltd.). Other sucrose fatty acid esters that can be used include, for example, Ryoto® Sugar Ester P1570 (product name, sucrose palmitate ester, HLB value: approximately 15 (catalog value), manufactured by Mitsubishi Chemical Foods Corporation), and Ryoto® Sugar Ester P1670 (product name, sucrose palmitate ester, HLB value: approximately 16 (catalog value)) (both manufactured by Mitsubishi Chemical Foods Corporation). Furthermore, 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 are product names, manufactured by Daiichi Kogyo Seiyaku Co., Ltd.).

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

[0052] The content range of sucrose stearate can be appropriately combined from the lower and upper limits mentioned above, but for example, it can be in the range of 1.0% by mass or more and 4.0% by mass or less, or for example, 1.0% by mass or more and 3.0% by mass or less, or for example, 1.0% by mass or more and 2.0% by mass or less, or for example, 1.2% by mass or more and 1.8% by mass or less, or for example, 1.4% by mass or more and 1.6% by mass or less.

[0053] Furthermore, when sucrose stearate is used alone as an emulsifier, or in combination with lysolecithin and / or polyglycerol fatty acid esters, the amount used can be specified by the above lower and upper limits. For example, when used alone, an amount of 1.0% by mass or more and 4.0% by mass or less may be preferred.

[0054] (Polyglycerin fatty acid ester) As the polyglycerol fatty acid ester, one or more known polyglycerol fatty acid esters can be used in appropriate combinations. When used alone or in combination with sucrose stearate ester and / or lysolecithin, these polyglycerol fatty acid esters may be advantageous in terms of median diameter, encapsulation, cell uptake, and appearance changes of the lipid components.

[0055] From the viewpoint of further improving emulsification stability, polyglycerin fatty acid esters have HLB values ​​that are, for example, 10 or higher, 11 or higher, 12 or higher, 13 or higher, 14 or higher, and 15 or higher. Alternatively, the HLB value may be 20 or lower, 18 or lower, or 17 or lower.

[0056] It is preferable that the HLB value of polyglycerin fatty acid esters differs from that of sucrose fatty acid esters. Different HLB values ​​may be advantageous for emulsification stability, encapsulation of lipid components, median diameter of lipid components, and cell uptake. It is preferable that the HLB value of polyglycerin fatty acid esters is lower than that of sucrose fatty acid esters, for example, by 1 to 4, 1 to 3, or 1 to 2. From this viewpoint, the HLB value of polyglycerin fatty acid esters is, for example, 11 to 13, or 12.

[0057] Examples of polyglycerin fatty acid esters include esters of polyglycerin 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. The unsaturated fatty acids having 10 to 22 carbon atoms include linear unsaturated fatty acids and branched unsaturated fatty acids, and the saturated fatty acids having 8 to 22 carbon atoms include linear saturated fatty acids and branched saturated fatty acids. Having the degree of polymerization of polyglycerin within the above range results in better emulsion stability.

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

[0059] Polyglycerol fatty acid esters may be monoesters or diesters, but monoesters are sometimes preferred.

[0060] While there are no particular limitations on the polyglycerin fatty acid esters, examples include decaglycerin laurate, decaglycerin myristic acid, decaglycerin palmitate, decaglycerin stearate, hexaglycerin monooleate, decaglycerin monooleate, decaglycerin linoleate, decaglycerin isostearate, hexaglycerin isostearate, decaglycerin monoisostearate, and the like. Among these, it is preferable that at least one is selected from the group consisting of decaglycerin laurate, decaglycerin myristic acid, decaglycerin palmitate, decaglycerin stearate, polyglyceryl isostearate, and polyglyceryl oleate. Among these, decaglycerin monooleate (HLB value: 12) and decaglycerin myristic acid (HLB value: 16) may be advantageous in some cases.

[0061] Commercially available polyglycerin fatty acid esters may be used, such as Poem J-0381V (manufactured by Riken Vitamin Co., Ltd.) and SY Glister MM-750 (manufactured by Sakamoto Pharmaceutical Co., Ltd.).

[0062] The content of polyglycerin fatty acid esters can be defined in relation to the total amount of oil components, sugars, and emulsifiers in the powder composition. For example, when used alone as an emulsifier, the content of polyglycerin fatty acid esters can be 6.0% by mass or more, or for example, 7.0% by mass or more, or for example, 8.0% by mass or more, or for example, 9.0% by mass or more. Alternatively, it can be 12% by mass or less, or for example, 11% by mass or less, or for example, 10% by mass or less. In such cases, the range of polyglycerin fatty acid esters can be set by appropriately combining the lower and upper limits described above, for example, 6.0% by mass or more and 12% by mass or less, or for example, 6.0% by mass or more and 11% by mass or less, or 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, or 9.0% by mass or more and 10% by mass or less.

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

[0064] (Lysolecithin) The powder composition can use lysolecithin in combination with sucrose stearate as an emulsifier. Lysolecithin is a hydrolyzed lecithin obtained by enzymatically treating lecithin, which is mainly composed of phospholipids that are major components of animal cell membranes, to unichain the alkyl chains and 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 oil and fat components when combined with sucrose stearate, or when combined with sucrose stearate and polyglycerol fatty acid esters. In this specification, lysolecithin includes so-called hydrogenated enzymatically hydrolyzed lecithin, which has been hydrogenated to improve oxidative stability by converting the bound fatty acids into saturated fatty acids.

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

[0066] The lysolecithin content may be defined in relation to the total amount of oil components, sugars, and emulsifiers in the powder composition. That is, the lysolecithin content may be, for example, 0.1% by mass or more of the total amount, or for example, 0.5% by mass or more, or for example, 1% by mass or more, or for example, 2% by mass or more, or for example, 2.5% by mass or more, or for example, 2.8% by mass or more, or for example, 2.9% by mass or more. Alternatively, the content may be 9% by mass or less, or for example, 8% by mass or less, or for example, 6% by mass or less, or for example, 5% by mass or less, or for example, 4.5% by mass or less, or for example, 4% by mass or less, or for example, 3.5% by mass or less, or for example, 3.2% by mass or less, or for example, 3.1% by mass or less. The lysolecithin content can be set by appropriately combining the lower and upper limits mentioned above, for example, 0.1% to 5% by mass, 0.5% to 4% by mass, 1% to 4% by mass, 2% to 4% by mass, etc.

[0067] As lysolecithin, for example, commercially available lysolecithin can be used. While not particularly limited, examples include SLP-White Lyso (trade name, manufactured by Tsuji Oil Co., Ltd.), SLP-White Lyso H (trade name, manufactured by Tsuji Oil Co., Ltd.), Benecoat® BMI-40L (trade name, manufactured by Kao Corporation), and Resimar® EL (trade name, manufactured by Riken Vitamin Co., Ltd.).

[0068] In some cases, it may be advantageous to use only sucrose stearate ester 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 has an HLB value of 15 or more and 16 or less, and the amount of sucrose fatty acid ester can be, for example, 1% to 3% by mass, 1.2% to 1.7% by mass, etc., relative to the total amount.

[0069] In some cases, it may be advantageous to use only polyglycerin fatty acid esters as emulsifiers for powder compositions. In this case, the polyglycerin fatty acid ester has an HLB value of 12 to 16, and the amount of polyglycerin fatty acid ester can be, for example, 5% to 10% by mass, 6% to 10% by mass, 7% to 10% by mass, 8% to 10% by mass, etc., relative to the total amount.

[0070] When using emulsifiers in powder compositions, sucrose stearate and lysolecithin may be advantageous in terms of median diameter, encapsulation, and cell uptake of oil components. In particular, sucrose stearate with an HLB value of 15 to 16 may be advantageous. In this case, lysolecithin can be present in amounts of, for example, 2% to 4% by mass or 2.5% to 3.5% by mass relative to the total amount, and sucrose stearate can be present in amounts of, for example, 1% to 3% by mass or 1.2% to 1.7% by mass.

[0071] Furthermore, it may be advantageous to combine sucrose stearate, lysolecithin, and polyglycerol fatty acid ester as emulsifiers used in the powder composition. In this case, it may be advantageous for the sucrose stearate to have an HLB value of 15 to 16, and for the polyglycerol fatty acid ester to have an HLB value of 11 to 13. In this case, the sucrose stearate can be in amounts of 1% to 3% by mass, 1.2% to 1.7% by mass, etc., and the polyglycerol fatty acid ester can be in amounts of 1% to 2% by mass, 1.2% to 1.7% by mass, etc. The lysolecithin can be in amounts of, for example, 2% to 4% by mass, 2.5% 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, but can be, for example, 1% by mass or more and 10% by mass or less, 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] (Antioxidant) The powder composition may contain an antioxidant. The antioxidant is not particularly limited, and one or more known antioxidants can be used as appropriate. Examples include at least one compound selected from the group consisting of ascorbic acid, ascorbic acid derivatives, and salts thereof, vitamin E such as tocopherol and its derivatives, vitamin A, retinoic acid, retinol, retinyl acetate, retinyl palmitate, retinyl acetate, retinyl palmitate, tocopheryl retinoic acid, vitamin C and its derivatives, kinetin, sesamin, alpha-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, rosehip extract, and rosemary extract.

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

[0075] (Other ingredients) In addition to the components described above, the powder composition may contain one or more other components as needed. The other components are not particularly limited, as long as they are suitable for use in 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 characteristics relating to the median diameter distribution of the oil components. Preferably, the characteristics are median diameter, cell uptakeability, and further, encapsulation rate and / or water solubility.

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

[0078] The volume-based median diameter of the oil component can be measured as the volume-based median diameter of the oil droplet using a laser diffraction / scattering particle size distribution analyzer (LA-960, Horiba, Ltd.) by dynamic light scattering. A sample prepared by dissolving 1 g of the powder composition in 5 mL of deionized water, and then dispersing 100 μl of this aqueous solution in 20 mL of deionized water, can be measured using the above-mentioned apparatus. This average particle diameter corresponds to the average particle diameter (volume-based median diameter) of the oil droplets in the oil droplet emulsion in water during the manufacturing process of the powder composition.

[0079] Sea buckthorn fruit oil contains various fatty acids. Examples include palmitoleic acid, palmitic acid, oleic acid, linoleic acid, alpha-linolenic acid, and stearic acid. In particular, palmitoleic acid is present in amounts of, for example, 10% by mass or more of the total amount of sea buckthorn fruit oil. This is because an amount of 10% by mass or more is convenient for ensuring an effective oral intake of palmitoleic acid. For example, it may be 15% by mass or more, 20% by mass or more, 25% by mass or more, or 30% by mass or more. There is no particular upper limit, but for example, it may be 35% by mass or less, or 30% by mass or less. The range of palmitoleic acid content can be set by appropriately combining the lower and upper limits mentioned above, 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, or 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, beta-carotene, lutein, zeaxanthin, ursolic acid, oleanolic acid, and other nutrients.

[0081] Lutein contributes to improving skin elasticity. The lutein content is, for example, 0.002% by mass or more, 0.003% by mass or more, 0.004% by mass or more, 0.005% by mass or more, and 0.006% by mass or more. A higher lutein content can be expected to lead to greater improvement in skin elasticity. There is no particular upper limit, but for example, it is 0.008% by mass or less, and for example, 0.007% by mass or less. The range of lutein content can be set by appropriately combining the lower and upper limits mentioned above, for example, 0.002% by mass or more and 0.008% by mass or less, and for example, 0.003% by mass or more and 0.007% by mass or less.

[0082] Zeaxanthin contributes to improving skin elasticity. The zeaxanthin content in sea buckthorn fruit oil is, for example, 0.010% by mass or more, 0.015% by mass or more, 0.020% by mass or more, 0.025% by mass or more, 0.030% by mass or more, 0.035% by mass or more, 0.040% by mass or more, and 0.050% by mass or more. Higher amounts of zeaxanthin can be expected to lead to greater improvement in skin elasticity. There is no particular upper limit, but for example, it is 0.080% by mass or less, 0.070% by mass or less, and 0.060% by mass or less. The range of zeaxanthin content can be set by appropriately combining the lower and upper limits mentioned above, 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] Beta-carotene contributes to improving skin elasticity. The beta-carotene content in sea buckthorn fruit oil is, for example, 0.010% by mass or more, 0.015% by mass or more, 0.020% by mass or more, and 0.025% by mass or more. A higher amount of beta-carotene can be expected to lead to greater improvement in skin elasticity. The upper limit is not particularly limited, but for example, it is 0.035% by mass or less, and for example, 0.030% by mass or less. The range of beta-carotene content can be set by appropriately combining the lower and upper limits mentioned above, for example, 0.010% by mass or more and 0.030% by mass or less, and 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 palmitoleic acid, which similarly 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 universally present in the bodies of animals, including humans.

[0085] Therefore, sea buckthorn fruit oil contains multiple active ingredients for improving skin elasticity derived from carotenoids and fatty acids, and its main component is fatty acids that contain excellent dissolving and delivery media for these active ingredients. As a result, it itself constitutes a useful skin elasticity improving composition.

[0086] In particular, sea buckthorn fruit oil contains palmitoleic acid as one of its main fatty acids, as well as zeaxanthin, beta-carotene, and lutein as its main carotenoids. All of these contribute to improving skin elasticity, and because they are present as major components, sea buckthorn fruit oil significantly contributes to improving skin elasticity. Therefore, sea buckthorn fruit oil is a useful active ingredient in skin elasticity improving agents. Skin elasticity improving agents contribute significantly to improving skin elasticity by containing palmitoleic acid and carotenoids such as zeaxanthin in the aforementioned amounts or within the specified range.

[0087] Sea buckthorn fruit oil is an effective active ingredient that can be used both topically and orally. Since skin elasticity improvement is a physical property expressed by skin tissue, even if cell-level experiments show promotion of the expression of the gene encoding COL1A, a precursor protein of collagen constituent proteins, and suppression of the expression of the gene encoding the collagen-degrading enzyme MMP1, it is unclear whether this improvement in skin elasticity occurs when ingested orally. However, according to the inventors, sea buckthorn fruit oil has been found to be linked to the promotion and suppression of collagen-related gene expression at the cellular level, resulting in improved skin elasticity when ingested orally. Therefore, sea buckthorn fruit oil contributes to improving skin elasticity whether used topically or orally. In particular, sea buckthorn fruit oil is considered useful for addressing skin elasticity loss caused by UV irradiation because it suppresses the expression of the gene encoding the collagen-degrading enzyme (MMP-1), which is enhanced by UV irradiation.

[0088] Skin elasticity improving agents are useful for improving the skin elasticity of animals such as humans. More specifically, they can improve the viscoelasticity of the skin. Skin viscoelasticity can be evaluated, for example, by the softness and shape recovery ability of the skin. By improving the viscoelasticity of the skin, skin elasticity improving agents also contribute to the prevention, improvement, or treatment of wrinkles and sagging.

[0089] Skin softness and skin resilience can be measured by the methods disclosed in the examples described later.

[0090] Skin elasticity improving agents can be used as ingredients in various foods and beverages to create foods and beverages (compositions) that can improve skin elasticity. The type of food and beverage is not particularly limited. Food and beverages can take the form of various ordinary foods and beverages, as well as health foods (so-called functional foods, health supplements, nutritionally fortified foods, nutritional supplements, etc.) and various forms of supplements (typically tablets, capsules). Capsules may be soft capsules or hard capsules.

[0091] Furthermore, skin elasticity improving agents can be used as raw materials for cosmetics, pharmaceuticals, and quasi-drugs to constitute topical skin compositions. Topical skin compositions can take various known forms such as lotions, emulsions, creams, gels, powders, pastes, and sprays. Those skilled in the art can appropriately select various components acceptable as cosmetics, pharmaceuticals, and quasi-drugs according to these forms and prepare these topical preparations.

[0092] Skin elasticity improving agents can be used as raw materials for pharmaceuticals intended for oral administration to constitute oral pharmaceutical compositions. Such pharmaceutical compositions can take various known forms, such as tablets, capsules, pills, powders, granules, liquids, etc. Those skilled in the art can appropriately select various pharmaceutically acceptable components according to these forms and prepare these oral pharmaceutical compositions. Typical oral pharmaceutical compositions are tablets or capsules (soft capsules and hard capsules).

[0093] Skin elasticity improving agents can also take the form of pharmaceutical compositions intended for parenteral administration, including local tissue administration, intradermal, subcutaneous, and intramuscular injections. Those skilled in the art can appropriately select various pharmacopoeia-acceptable components according to these forms and prepare these parenteral pharmaceutical compositions.

[0094] In these various compositions, the content of sea buckthorn fruit oil is determined as appropriate. Based on the total amount of the composition, for example, it may be 1% by mass or more, 10% by mass or more, 15% by mass or more, or 20% by mass or more. The upper limit is not particularly limited, but based on the total amount of the composition, for example, it may be 40% by mass or less, 35% by mass or less, 30% by mass or less, or 25% by mass or less. In the composition, the range of sea buckthorn fruit oil content can be set by appropriately combining these lower and upper limits, for example, 1% by mass or more and 40% by mass or less, 10% by mass or more and 30% by mass or less, or 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 in this specification 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. The promotion of COL1A gene expression means the promotion of 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 can be accessed at the NIH website (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. An inhibitor of MMP-1 gene expression means inhibition of the degradation of collagen that contributes to skin elasticity. 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] <Manufacturing of sea buckthorn fruit oil> After washing the sea buckthorn fruits with cold water, they were juiced, and the peel and seeds were removed. The juice was centrifuged to separate the oil from the fruit juice. The oil was filtered and then vacuum-dried to obtain sea buckthorn fruit oil. The obtained sea buckthorn fruit oil contained 29.1% by mass of palmitoleic acid, 0.0045% by mass of lutein, and 0.063% by mass of zeaxanthin. [Examples]

[0103] <Effect of sea buckthorn fruit oil on promoting collagen (COL1A1) gene expression> Normal human dermal fibroblasts were cultured in 5% FBS-containing DMEM medium at 5% CO2 and 37°C until confluence. After changing the medium and adding 1 ppm sea buckthorn fruit oil, 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 as the control instead of sea buckthorn fruit oil. 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 culture medium. This indicates that sea buckthorn fruit oil promotes the expression of genes related to collagen synthesis. [Examples]

[0105] <Effect of UV irradiation on the suppression of collagen-degrading enzyme (MMP-1) gene expression by sea buckthorn fruit oil> Normal human dermal fibroblasts were cultured in 5% FBS-containing DMEM medium at 5% CO2 and 37°C until confluence. After changing the medium and adding sea buckthorn fruit oil to a concentration of 1 ppm, the cells were cultured for 24 hours, washed with HBSS buffer, and treated with 100 mJ / cm³. 2 The cells were irradiated with UV-B light. After UV-B irradiation, RNA was extracted from cells cultured for 24 hours in DMEM medium containing 0.5% FBS, and the gene expression of collagenase (MMP-1) was analyzed by real-time RT-PCR. Dimethyl sulfoxide (DMSO) was used as the control instead of sea buckthorn fruit oil. The results are shown in Figure 2.

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

[0107] <Effect of UV irradiation on suppressing collagen-degrading enzyme (MMP-1) gene expression by carotenoids and palmitoleic acid> Normal human dermal fibroblasts were cultured in 5% FBS-containing DMEM medium at 5% CO2 and 37°C until confluence. After changing the medium, the main carotenoids (zeaxanthin, β-carotene, and lutein) contained in 1 ppm of sea buckthorn fruit oil were added at 0.63 ppb each, or palmitoleic acid was added at 0.291 ppm. After 24 hours of culture, the cells were washed with HBSS buffer and treated with 100 mJ / cm³. 2The cells were irradiated with UV-B light. Note that 0.63 ppb corresponds to the zeaxanthin content in the sea buckthorn fruit oil used in Example 2. Also, 0.291 ppb corresponds to the palmitoleic acid content in the sea buckthorn fruit oil. After UV-B irradiation, RNA was extracted from cells cultured for 24 hours in 0.5% FBS-containing DMEM medium, and the gene expression of collagenase (MMP-1) was analyzed by real-time RT-PCR. Dimethyl sulfoxide (DMSO) was used as the control instead of sea buckthorn fruit oil. The results are shown in Figure 3. The letters in the graph indicate statistically significant differences. (P<0.05, n=3)

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

[0109] <Skin elasticity improvement effect of sea buckthorn fruit oil> [1] Manufacturing of test foods and comparative 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, and the mixture was prepared so that sea buckthorn fruit oil constituted 20% by mass of the total solids.

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

[0111] Next, this liquid was treated in a nitrogen-filled chamber using a homomixer (TK Homomixer, manufactured by Primix Corporation) at a rotation speed of 8,000 rpm for 60 minutes. Then, this treated liquid was further treated in three passes using a high-pressure homogenizer (NanoVeta C-ES, manufactured by Yoshida Machinery Industry Co., Ltd.) at a processing pressure of 100 MPa to obtain an oil-in-water emulsion.

[0112] The volume-based median diameter of the oil component in an 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 above oil-in-water emulsion in 5 mL of deionized water, and then diluting 100 μL of this solution with 20 mL of deionized water. The resulting sample was analyzed and found to be 140 nm.

[0113] This oil-in-water emulsion was spray-dried using a spray drying apparatus (CL-8, manufactured by Okawara Chemical Machinery Co., Ltd.) at a tower temperature of 140°C and an atomizer rotation speed of 20Hz 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 according to the formulations shown in Table 1 to produce Test Food 1 and Test Food 2. Test Food 1 contains 9 mg of sea buckthorn fruit oil per capsule, and Test Food 2 contains 27 mg of sea buckthorn fruit oil per capsule. The control food contains 27 mg of soybean oil per capsule.

[0115] [Table 1]

[0116] [2] Double-blind study on improvement of skin elasticity using 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 and 2, and a control food for four weeks improved skin elasticity.

[0117] (1) Research design The study was conducted over a four-week period from late January to early March 2023 using a randomized, placebo-controlled, double-blind, parallel-group comparative study design. Note that this period is seasonally dry, and typically, skin elasticity and resilience tend to be reduced (see comparison food intervention group).

[0118] (2) Allocation The 99 participants were randomly assigned to three groups: Test Food 1 intervention group, Test Food 2 intervention group, and Control Food intervention group, each consisting of 33 participants. Table 2 shows the characteristics of the participants in each 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 instructed not to be able to distinguish between the three individuals based on their appearance, taste, or smell, and to take four hard capsules daily with water at any time of their choosing. • Intervention period: 4 weeks

[0121] (4) Evaluation items and measurements The evaluation item was skin viscoelasticity, measured before ingestion and 4 weeks after ingestion using a Cutometer dual® MPA 580 (Courage+Khazaka electronic GmbH, Germany). The measurement procedure involved placing the Cutometer's measuring probe on the subject's left cheek, applying negative pressure to the skin through the probe's opening for a certain period of time, and then releasing the pressure. The maximum change during negative pressure suction was defined as R0 (in mm), and the change after the negative pressure was released was defined as R8 (in mm).

[0122] R0 indicates the firmness (softness) of the skin, and R8 indicates the skin's ability to return to its original state. Higher values ​​for both indicators indicate better condition.

[0123] (5) Statistical methods All statistical analyses were performed using two-tailed tests, with a significance level of 5%. SPSS Statistics version 23 or later was used, and other validated statistical software was used as needed. All pairwise comparisons were performed against the control food intervention group.

[0124] (6) Results (6-1) Skin viscoelasticity measurement Figures 4 and 5 show the Mean, SD, Med, Min, Max, group differences and their SE, 95% CI-, 95% CI+, and statistical analysis results for skin viscoelasticity measurements. Figure 4 shows the analysis results for flexibility (R0), and Figure 5 shows the analysis results for resilience (R8).

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

[0126] Furthermore, as shown in Figure 5, regarding skin regeneration, after 4 weeks of ingestion, the total recovery (R8) of the first curve of the left cheek was significantly higher in the Test Food 1 intervention group (Test Food 1 intervention group: 0.1548±0.1095 mm, between-group difference: 0.0398 mm [95% 0.0006, 0.0791], P=0.047) and the Test Food 2 intervention group (Comparative food intervention group: 0.1160±0.0372 mm, Test Food 2 intervention group: 0.1704±0.1514 mm, between-group difference: 0.0533 mm [0.0141, 0.0926], P=0.008) compared to the control food intervention group.

[0127] Furthermore, the mean values ​​of R0 and R8 in the control food intervention groups from before 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 all indicators decreased during the study period. From this, it was concluded that in this study, the intervention groups of test food 1 and test food 2 suppressed the decrease in skin flexibility and shape recovery during the study period. [Industrial applicability]

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

Claims

1. A powder composition for improving skin elasticity for oral intake, Oil derived from sea buckthorn pulp, Sugars and, Emulsifier and, It contains, The oil contains, with respect to the total amount of the oil, palmitoleic acid in an amount of 10% to 35% by mass and zeaxanthin in an amount of 0.010% to 0.080% by mass. The composition comprises oil and fat particles, each having a volume-based median diameter of 100 nm or more and 400 nm or less, and is an orally ingestible powder composition for improving skin elasticity.

2. The skin elasticity improving powder composition according to claim 1, wherein the volume-based median diameter is 100 nm or more and 250 nm or less.

3. The skin elasticity improving powder composition according to claim 1, which has an effect of promoting the expression of the collagen-related gene COL1A1 and an effect of suppressing the expression of the MMP-1 gene.

4. A skin elasticity improving powder composition according to any one of claims 1 to 3, which significantly improves skin flexibility or resilience after four weeks of oral intake.

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