WO2015083630A1 - コラーゲンペプチド、エラスチンペプチドおよびプロテオグリカンを含む組成物 - Google Patents
コラーゲンペプチド、エラスチンペプチドおよびプロテオグリカンを含む組成物 Download PDFInfo
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- WO2015083630A1 WO2015083630A1 PCT/JP2014/081505 JP2014081505W WO2015083630A1 WO 2015083630 A1 WO2015083630 A1 WO 2015083630A1 JP 2014081505 W JP2014081505 W JP 2014081505W WO 2015083630 A1 WO2015083630 A1 WO 2015083630A1
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- peptide
- collagen
- proteoglycan
- elastin
- skin
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- 238000000746 purification Methods 0.000 description 1
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Classifications
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
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- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
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- C08L89/00—Compositions of proteins; Compositions of derivatives thereof
- C08L89/04—Products derived from waste materials, e.g. horn, hoof or hair
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- A—HUMAN NECESSITIES
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- A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
Definitions
- the present invention relates to a composition comprising a collagen peptide, an elastin peptide and a proteoglycan. More specifically, the present invention relates to a composition having an effect of improving skin beauty, wherein the weight of collagen peptide and the total weight of elastin peptide and proteoglycan are in a predetermined ratio.
- the skin is composed of the stratum corneum, epidermis, basement membrane and dermis from the outside.
- the dermis is the largest part of the area, and the cells are not packed as densely as the epidermis. Rather, the extracellular space is wide and filled with a network of macromolecules called “extracellular matrix”.
- This extracellular matrix is directly related to skin elasticity, elasticity, freshness, metabolism, etc.
- Skin aging symptoms such as wrinkles and sagging are the dermis whose main components are fibroblasts and extracellular matrix. It is known to occur in connective tissue fibers.
- Fibrous proteins such as collagen and elastin that make up the extracellular matrix and polysaccharides called acidic mucopolysaccharides such as hyaluronic acid and dermatan sulfate are produced by fibroblasts. Promoting contributes to preventing and improving wrinkles and sagging by giving the skin a gloss and gloss. From such a viewpoint, various beverages and cosmetics containing components having a fibroblast proliferation promoting action have been proposed.
- collagen has a bone strengthening action (patent document 1) that leads to prevention and improvement of osteoporosis, a metabolism promoting action of the living tissue that improves a decrease in function of the living tissue with aging (patent document 2), a skin metabolism promoting action, Physiological effects such as skin activation (Patent Document 3) and skin aging prevention (Patent Document 4) for the purpose of prevention and improvement of wrinkles have been found. Widely used as biofunctional materials.
- Non-patent Document 1 Collagen degradation products are known to be decomposed into amino acids, dipeptides or tripeptides in the process of digestion and absorption, and it has been shown that among these, dipeptides or tripeptides are effective.
- JP-A-9-255588 Japanese Patent Laid-Open No. 7-278012 JP-A-9-67262 JP 2005-314265 A Japanese Patent No. 4996155
- an object of the present invention is to provide a composition capable of exerting a useful skin cosmetic improvement effect even when the intake amount of collagen peptide is reduced.
- the present invention includes, but is not limited to, the following aspects of the invention.
- a composition comprising a collagen peptide, an elastin peptide and a proteoglycan, wherein the weight ratio of the collagen peptide to the total weight of the elastin peptide and the proteoglycan [(collagen peptide) :( elastin peptide + proteoglycan)] is 1 : The above composition, which is 10 to 1: 0.0000001.
- the composition according to (1) wherein the weight ratio [(collagen peptide) :( elastin peptide)] of the weight of collagen peptide to the weight of elastin peptide is 1: 5 to 1: 0.000001.
- composition according to (1) wherein the weight ratio [(collagen peptide) :( proteoglycan)] between the weight of the collagen peptide and the proteoglycan is 1: 0.2 to 1: 0.0000001.
- a skin improver comprising a collagen peptide, an elastin peptide and a proteoglycan, wherein the weight ratio of the weight of the collagen peptide to the total weight of the elastin peptide and the proteoglycan [(collagen peptide) :( elastin peptide + proteoglycan)]
- a skin-improving agent having a ratio of 1:10 to 1: 0.0000001.
- the skin improving agent according to (6) comprising a fibroblast proliferation promoting action.
- composition that effectively exhibits a skin beauty improving effect even with a low concentration of collagen peptide.
- composition of the present invention is also advantageous in that the amount of collagen peptide exhibiting an unfavorable taste can be suppressed.
- the composition which is one embodiment of the present invention includes a collagen peptide, an elastin peptide, and a proteoglycan as active ingredients.
- the composition of the present invention has an effect of promoting the growth of fibroblasts existing in living tissue, particularly in the dermal tissue of the skin, and thereby has an effect of improving skin beauty.
- the collagen peptide used in the present invention can be obtained by subjecting denatured collagen such as collagen or gelatin to a hydrolysis treatment with enzymes, acids, alkalis, etc., or artificially synthesized These may be used, and one or more of these may be used.
- denatured collagen such as collagen or gelatin
- enzymes, acids, alkalis, etc. or artificially synthesized These may be used, and one or more of these may be used.
- collagen and gelatin for example, those derived from animals such as cattle, pigs, chickens, and fish, especially collagen proteins extracted from connective tissues such as animal skin, bones and tendons, fish skin, and fish scales are used. Can do.
- the enzyme used for the preparation of the collagen peptide may be any enzyme that can cleave the peptide bond of collagen or gelatin.
- the acid for example, hydrochloric acid, sulfuric acid, nitric acid and the like can be used.
- alkali examples include sodium hydroxide and calcium hydroxide.
- an aqueous solution of a hydrolyzed collagen peptide may be used as it is, or a powdered powder by drying or the like may be used.
- collagen peptides of any molecular weight can be used, but for example, the molecular weight is 10,000 or less, for example, 100 to 7,000, preferably 100 to 5,000.
- the molecular weight of the collagen peptide can be measured by a known quantitative method such as HPLC or gel filtration.
- collagen peptides generally have lower absorbability in the body as the molecular weight increases. Absorbability in the body increases as the molecular weight decreases, but unfavorable tastes (bitterness, sardine, etc.) peculiar to peptides occur. End up.
- the combined use of elastin peptide and proteoglycan makes it possible to keep the amount of collagen peptide used low. Therefore, even when a collagen peptide having a relatively low molecular weight with good absorbability in the body is used, Undesirable taste problems are less likely to occur.
- collagen peptide either an extract or a purified product may be used, but a collagen peptide having a purity of 50% by weight or more, more preferably 70% by weight or more, further preferably 90% by weight or more is used.
- collagen peptide a commercially available product may be used.
- “collagen peptide SCP-5100” manufactured by Nitta Gelatin Co., Ltd.) or the like can be used.
- the collagen peptide content in the composition of the present invention is difficult to uniformly define depending on the type and content of the concomitant raw materials, but is 1% to 99.9% by weight, preferably 30% to 99.9% by weight. More preferably, it is 50 to 90% by weight. If the content of the collagen peptide is less than 1% by weight or more than 99.9% by weight, the desired effect of the present invention cannot be expected.
- the elastin peptide in the present invention means a water-soluble elastin peptide.
- elastin peptides include those extracted from animal biological tissues such as cattle, pigs, chickens, sheep, and fish, or by subjecting water-soluble or insoluble elastin to hydrolysis with enzymes, acids, alkalis, etc.
- the resulting decomposition product can be used, or an artificially synthesized product can be used, or one or more of these can be used.
- the molecular weight of the elastin peptide used in the present invention is not particularly limited, and elastin having any molecular weight can be used.
- Either an extract or a purified product may be used as the elastin peptide, but preferably, the purity is 50% by weight or more, more preferably 70% by weight or more, and further preferably 90% by weight or more.
- the elastin peptide a commercially available product may be used. For example, “Bitsu Elastin FI” (manufactured by Nihon Suisan Co., Ltd.), “Tuna Elastin HS-1” (manufactured by Hagoromo Foods Co., Ltd.), Sangyo Co., Ltd.) can be used.
- the content of the elastin peptide in the composition of the present invention varies depending on the type and content of the concomitant raw materials such as collagen peptide, but is 0.0000002% by weight to 10% by weight, preferably 0.000002% by weight to 1% by weight. More preferably, it is 0.000004 wt% to 0.2 wt%.
- proteoglycan used in the present invention is not particularly limited, and for example, those extracted from animal cartilage such as salmon, whale, salmon and ray can be used.
- a proteoglycan having a molecular weight of 10,000 to 10,000,000, preferably 100,000 to 1,000,000 can be used.
- proteoglycan either an extract or a purified product may be used, but preferably a proteoglycan having a purity of 5% by weight or more, more preferably a purity of 10% by weight or more, and further preferably a purity of 15% by weight or more.
- a commercially available proteoglycan may be used.
- the content of proteoglycan in the composition of the present invention varies depending on the type and content of the concomitant raw materials such as collagen peptides, but is 0.0000001% to 5% by weight, preferably 0.000001% to 0.5% by weight, More preferably, it is 0.000002 wt% to 0.1 wt%.
- the weight ratio [(collagen peptide) :( elastin peptide + proteoglycan)] of the weight of the collagen peptide in the composition to the total weight of the elastin peptide and proteoglycan is, for example, 1:10 to 1: 0. 0000001, preferably 1: 5 to 1: 0.000001, more preferably 1: 2 to 1: 0.00001.
- the present inventors have obtained the knowledge that the effect of improving the skin is higher by blending the collagen peptide in combination with the elastin peptide and proteoglycan as compared with the case where the same dose of the collagen peptide alone is used.
- the weight ratio [(collagen peptide) :( elastin peptide)] of the weight of collagen peptide and the weight of elastin peptide in the composition of the present invention is, for example, 1: 5 to 1: 0.000001, preferably Is from 1: 2 to 1: 0.00001, more preferably from 1: 1 to 1: 0.0001, and the weight ratio of the weight of collagen peptide to the weight of proteoglycan in the composition [(collagen peptide) :( proteoglycan )] Is, for example, 1: 0.2 to 1: 0.0000001, preferably 1: 0.1 to 1: 0.0000005, more preferably 1: 0.02 to 1: 0.000002. .
- the present inventors have also obtained, through experiments, an unexpected finding that a particularly high collagen production promoting effect is exhibited when an elastin peptide and proteoglycan are blended at a low ratio with respect to a collagen peptide. That is, when the same amount of collagen peptide is ingested, a higher skin improvement effect can be obtained when the weight ratio of elastin peptide and proteoglycan in the composition is low. Therefore, according to the composition of the present invention, the amount of the active ingredient can be kept very low, it is possible to provide a composition that is easy to ingest and is excellent in cost.
- additives can be blended in the composition of the present invention.
- Additives are not particularly limited, but those commonly used for oral ingestion are preferred, such as excipients, binders, disintegrants, lubricants, preservatives, corrigents, flavoring agents, coloring agents, fragrances, etc. Additives can be used, and materials with a known skin improvement effect can be used.
- composition according to the present invention can be combined with other beauty / health ingredients known to have a skin improvement effect.
- other beauty / health ingredients known to have a skin improvement effect.
- ceramide, lactic acid bacteria, vitamins (vitamin C etc.), minerals (calcium etc.), a plant extract, etc. are mentioned.
- composition according to the present invention may be blended with components known to have a fibroblast proliferation promoting action, a collagen production promoting action, and a skin aging preventing action exemplified below.
- the composition of the present invention is preferably an oral composition.
- food / beverage products, a pharmaceutical, a quasi-drug, etc. are contained, It is preferable that it is food / beverage products, and it is more preferable that it is a drink.
- foods and drinks include health foods, dietary supplements, foods for specified health use, supplements, dairy products, soft drinks, powders for dissolution at the time of use, and pet foods and drinks, livestock feeds, and the like.
- a powder for dissolution at the time of use is preferable.
- the powder for dissolution at the time of use can be further dissolved in coffee, tea, juice, yogurt, soup, etc., or mixed with food.
- the powder for use in daily use is particularly excellent in terms of stability during storage and ease of administration in that it can be taken after being dissolved or added to a favorite one.
- it can be used in the form of tablets and capsules.
- the forms of pharmaceuticals and quasi drugs are typically oral preparations such as granules, tablets, capsules and liquids.
- the dose of the composition of the present invention can be determined as appropriate based on the age, weight, health status, etc. of the subject.
- the daily intake of a human adult is 10 mg to 100,000 mg, preferably 500 mg. mg to 15,000 mg, more preferably 3,000 mg to 10,000 mg, which can be taken and administered in a single dose or divided into multiple doses. With the lowering of the collagen peptide dose, it has become possible to reduce the daily intake of the composition.
- composition according to the present invention can be used for the prevention and improvement of skin symptoms.
- skin moisturizing function and elasticity decrease, skin tension and gloss decrease, roughness, wrinkles, dullness, etc. Can be prevented or improved.
- the effect of improving skin is enhanced by the action of promoting the growth of fibroblasts present in living tissues, particularly the dermal tissue of the skin, and the action of promoting collagen production and the action of preventing skin aging associated with the action of promoting the proliferation of fibroblasts. It is known to be demonstrated. Therefore, as another aspect of the present invention, the composition has a fibroblast proliferation promoting action, a collagen production promoting action, or a skin aging preventing action.
- the fibroblast proliferation promoting action means an action of promoting the growth of fibroblasts present in living tissue, particularly skin dermal tissue, and can be evaluated by a known method.
- the active ingredients of the fibroblast proliferation promoting action are collagen peptides, elastin peptides and proteoglycans. These components may be in the form of a solid, paste or liquid, which may be used as it is as a skin improving agent having a fibroblast proliferation promoting action, but if necessary, a known additive having a fibroblast proliferation promoting action In combination, it can be contained by a conventional method to prepare a composition.
- the collagen production enhancing action refers to the action of enhancing the production of collagen in the living body, and the production of collagen is more preferably the production of collagen by fibroblasts in the dermal tissue.
- Collagen production can be evaluated, for example, by measuring the amount of type I collagen produced by a known method, and the expression level of the collagen synthesis gene (COL1a1 gene) in human skin fibroblasts can be determined by a known method. It can also be confirmed by evaluation.
- Skin aging prevention means prevention of skin aging symptoms, and skin aging symptoms are selected from the group consisting of wrinkles, spots, dullness, freckles, sagging, open pores, roughness and rough skin. It contains at least one symptom.
- the present inventors In combination with the collagen peptide, the elastin peptide, and the proteoglycan, the present inventors have further promoted the proliferation of human skin fibroblasts as compared to the case where the collagen peptide alone is used. It has been confirmed by experiments that the expression is also enhanced, and it has also been confirmed that the production of collagen is significantly promoted. Based on these findings, those skilled in the art have found that even if the amount of collagen peptide intake is lower than before, a combination of collagen peptide, elastin peptide and proteoglycan can provide a significant skin improvement effect. Of course you can understand if there is.
- the composition of the present invention can be produced according to a conventional method, but the active ingredients collagen peptide, elastin peptide and proteoglycan may be blended as they are, or may be blended in a solid, paste or liquid form. Good. Moreover, other components, such as a well-known component and additive provided with a skin improvement effect, are mix
- collagen peptides for example, collagen peptides, elastin peptides and proteoglycans, and if necessary, excipients such as glucose (dextrose), dextrin, lactose, starch or processed products thereof, cellulose powder, vitamins, minerals, fats and oils of animals, plants and fish, White (proteins derived from plants and animals, hydrolysates thereof), sugars, pigments, fragrances, antioxidants, surfactants, other food additives, various nutritional functional ingredients, powders of ceramide, casein, etc.
- excipients such as glucose (dextrose), dextrin, lactose, starch or processed products thereof, cellulose powder, vitamins, minerals, fats and oils of animals, plants and fish, White (proteins derived from plants and animals, hydrolysates thereof), sugars, pigments, fragrances, antioxidants, surfactants, other food additives, various nutritional functional ingredients, powders of ceramide, casein, etc.
- Rukoto can Mixed with edible materials such as extracts and processed into powders, granules, pellets, tablets, etc., processed into foods and drinks of the above examples by conventional methods, and mixed liquid products with gelatin and sodium alginate , Coated with a coating agent such as carboxymethyl cellulose to form capsules, or processed into beverage (drinks) forms Rukoto can.
- a coating agent such as carboxymethyl cellulose to form capsules, or processed into beverage (drinks) forms Rukoto can.
- it is preferably processed into a nutritional supplement or health food, and more preferably processed into a powder form for dissolution at the time of use.
- a skin improvement agent comprising a collagen peptide, an elastin peptide and a proteoglycan in a predetermined ratio
- the weight ratio [(collagen peptide) :( elastin peptide + proteoglycan)] of the weight of the collagen peptide in the skin improving agent and the total weight of the elastin peptide and proteoglycan is, for example, 1:10 to 1: 0.0000001.
- the ratio is preferably 1: 5 to 1: 0.000001, more preferably 1: 2 to 1: 0.00001.
- the weight ratio [(collagen peptide) :( elastin peptide)] between the weight of the collagen peptide and the weight of the elastin peptide is, for example, 1: 5 to 1: 0.000001, preferably 1: 2 to 1: It is 0.00001, more preferably 1: 1 to 1: 0.0001. Furthermore, the weight ratio [(collagen peptide) :( proteoglycan)] between the weight of the collagen peptide and the proteoglycan is, for example, 1: 0.2 to 1: 0.0000001, preferably 1: 0.1 to It is 1: 0.0000005, more preferably 1: 0.02 to 1: 0.000002.
- the active ingredient contained in the skin improving agent, other components, the form and dose of the skin improving agent, the action / effect, and the production method can apply the contents described for the composition.
- the “skin improving agent” as used in the present invention means a preparation for preventing or improving symptoms such as a decrease in skin moisturizing function and elasticity, a decrease in skin tension and gloss, roughness, wrinkles and dullness.
- prevention / improvement of various skin symptoms by the skin improving agent may be accompanied by one or more actions selected from a fibroblast proliferation promoting action, a collagen production promoting action, and a skin aging preventing action.
- Example 1 Examination of the proliferation promoting action of human skin fibroblasts The effect of the fibroblast proliferation promoter of the present invention on the proliferation of skin fibroblasts was examined by the following method.
- ⁇ Preparation of test sample> Collagen peptide, elastin peptide and proteoglycan are weighed, dissolved in sterilized water, filtered and sterilized with a membrane filter having a pore size of 0.2 ⁇ m, and then 100 times with DMEM medium containing 0.1% FBS so that the sample concentrations shown in Table 1 below are obtained. Samples 2-6 were prepared by dilution. As a control, Sample 1 in which sterilized water was diluted 100-fold with a DMEM medium containing 0.1% FBS was used. In addition, the density
- Collagen peptide Nitta Gelatin Co., Ltd., molecular weight 100-1800, purity 100% Elastin peptide: “Bitsu Elastin FI” (Nihon Suisan Co., Ltd.), 100% purity Proteoglycan: “Proteoglycan F” (manufactured by Ichimaru Falcos), molecular weight: about 450,000, purity 20%
- ⁇ Evaluation method> Adult-derived normal human dermal fibroblast NHDF strain (purchased from Kurabo Corp.) was inoculated into Dulbecco's modified MEM medium (DMEM, manufactured by Nissui) containing 10% bovine serum (10%), and 10, The cells were seeded at 000 cells / well and cultured overnight at 37 ° C. and 5% CO 2. The test sample was dissolved in water, diluted 100-fold with 0.1% FBS-containing DMEM medium as described above, and added to the cells. Cell viability was measured 24 hours after each test sample (200 ⁇ l) was added. The cell viability was calculated from the amount of formazan produced by reduction with intracellular dehydrogenase using MTS reagent (Promega).
- the cell viability of each test sample was determined as a value (%) when the cell viability of the sample to which only the DMEM medium containing 0.1% FBS containing 1% water was added was taken as 100%. Formazan was measured by ultraviolet absorption at 490 nm. The inventor previously examined the FBS concentration at which the cells do not grow or decrease, confirmed that this condition was satisfied when cultured in 0.1% FBS-containing DMEM medium, and set the serum concentration of the test medium to 0. By adjusting to 1%, it was decided that the fibroblast proliferation promoting effect of only the test sample could be quantitatively evaluated.
- Example 2 Examination of collagen production promoting action of human skin fibroblasts ⁇ Preparation of test sample> According to the method of Example 1, Samples 1 to 7 shown in Table 2 below were prepared.
- ⁇ Evaluation method> The NHDF strain was ingested into a DMEM medium containing 10% FBS, seeded at 125,000 cells / well in a 24-well plate, and cultured overnight at 37 ° C. and 5% CO 2. The test sample was dissolved in water, diluted 100-fold with 0.1% FBS-containing DMEM medium, and added to the cells. Collagen production was measured 72 hours after adding each test sample. Collagen Type I, Human, ELISA Kit, manufactured by Acer Corporation) was used to quantify the type I collagen secreted into the cell culture supernatant. Color was developed using a polyclonal antibody specific for type I atelocollagen and measured by ultraviolet absorption at 450 nm.
- the collagen production amount of each test sample was determined as a value (%) when the collagen production amount of a sample to which only 0.1% FBS-containing DMEM medium containing 1% water was added was defined as 100%.
- the effect on collagen synthesis was kept low by lowering the serum concentration in the culture solution as in Example 1.
- Example 3 Influence on expression of various genes in human skin fibroblasts ⁇ Preparation of test sample> According to the method of Example 1, samples 1 to 4 shown in Table 3 below were prepared.
- NHDF strain was inoculated in DMEM medium containing 10% FBS, seeded at a density of 10,000 cells / well in a 96-well plate, and cultured overnight at 37 ° C. and 5% CO 2 .
- the test sample was dissolved in water, diluted 100-fold with 0.1% FBS-containing DMEM medium, and added to the cells. 24 hours after adding each test sample, the expression of type I collagen synthesis gene (Col1a1) was quantified by quantitative real-time PCR. ⁇ -actin was used as a control housekeeping gene.
- the cells were washed with PBS, and cDNA was prepared according to the protocol of TaqMan Gene Expression Cell-to-CT Kit (Ambion).
- Powder for dissolution at time of use Powders for dissolution at time of use (6.5 g) were prepared by weighing collagen, elastin, proteoglycan and dextrin as shown in Table 4 below and mixing them uniformly. When the obtained powder was dissolved in water, fruit juice, or consomme soup, they all had good dispersibility and were suitable as beverages to be used at the time of use.
- Formulation Example 2 Capsule In the soft capsule skin composed of the following components, the above-mentioned powder for dissolution for use was filled by a conventional method to obtain a soft capsule.
- Formulation Example 3 Tablet A mixture having the composition shown below was granulated and molded by a conventional method to obtain a tablet.
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Abstract
Description
(1)コラーゲンペプチド、エラスチンペプチドおよびプロテオグリカンを含む組成物であって、コラーゲンペプチドの重量とエラスチンペプチドおよびプロテオグリカンの合計重量との重量比[(コラーゲンペプチド):(エラスチンペプチド+プロテオグリカン)]が、1:10~1:0.0000001である、上記組成物。
(2)コラーゲンペプチドの重量とエラスチンペプチドの重量との重量比[(コラーゲンペプチド):(エラスチンペプチド)]が、1:5~1:0.000001である、(1)に記載の組成物。
(3)コラーゲンペプチドの重量とプロテオグリカンの重量との重量比[(コラーゲンペプチド):(プロテオグリカン)]が、1:0.2~1:0.0000001である、(1)に記載の組成物。
(4)経口用である、(1)~(3)のいずれかに記載の組成物。
(5)飲食品である(1)~(3)のいずれかに記載の組成物。
(6)コラーゲンペプチド、エラスチンペプチドおよびプロテオグリカンを含む皮膚改善剤であって、コラーゲンペプチドの重量とエラスチンペプチドおよびプロテオグリカンの合計重量との重量比[(コラーゲンペプチド):(エラスチンペプチド+プロテオグリカン)]が、1:10~1:0.0000001である、皮膚改善剤。
(7)線維芽細胞増殖促進作用を備える、(6)に記載の皮膚改善剤。
(8)コラーゲン産生促進作用を備える、(6)または(7)に記載の皮膚改善剤。
(9)皮膚老化防止作用を備える、(6)~(8)のいずれか1項に記載の皮膚改善剤。
<有効成分>
まず、本発明の一態様である組成物は、コラーゲンペプチド、エラスチンペプチドおよびプロテオグリカンを有効成分として含むことを特徴とする。本発明の組成物は、生体組織、とりわけ皮膚の真皮組織中に存在する線維芽細胞の増殖を促進させる作用を有することにより、皮膚の美容改善効果を奏するものである。
本発明において用いるコラーゲンペプチドは、コラーゲン、あるいはゼラチン等の変性コラーゲンを、酵素、酸、アルカリ等による加水分解処理等を施すことにより得られたものを用いることができ、また人工的に合成したものを用いてもよく、これらの1種または2種以上を用いることもできる。コラーゲンやゼラチンとしては、例えば、牛、豚、鶏等の動物由来や魚由来のもの、特に、動物の皮膚、骨および腱等の結合組織や、魚皮、魚鱗から抽出したコラーゲンタンパク質を用いることができる。
本発明におけるエラスチンペプチドは、水溶性エラスチンペプチドを意味する。エラスチンペプチドとしては、例えば、牛、豚、鶏、羊、魚等の動物性生体組織から抽出したものや、水溶性あるいは不溶性のエラスチンに酵素、酸、アルカリ等で加水分解処理等を施すことにより得られた分解物を使用することができ、また人工的に合成したものを用いてもよく、これらの1種または2種以上を用いることもできる。
本発明において用いるプロテオグリカンは特に限定されないが、例えば、鮫や鯨、鮭、エイ等の動物軟骨から抽出したものを使用することができる。
本発明においては、組成物中のコラーゲンペプチドの重量とエラスチンペプチドおよびプロテオグリカンの合計重量との重量比[(コラーゲンペプチド):(エラスチンペプチド+プロテオグリカン)]は、例えば、1:10~1:0.0000001であり、好ましくは1:5~1:0.000001、より好ましくは1:2~1:0.00001である。本発明者らは、コラーゲンペプチドを、エラスチンペプチドおよびプロテオグリカンと組み合わせて配合することにより、同用量のコラーゲンペプチド単体を使用した場合と比較して、皮膚改善効果が高いという知見を得ている。したがって、上記範囲の重量比で有効成分を含む組成物を摂取した場合、同用量のコラーゲンペプチドを摂取した場合と比較して、より高い皮膚改善効果を与えることが期待できる。また、より低濃度のコラーゲンペプチドでも有効に皮膚改善効果を示すことから、上記範囲の重量比で有効成分を含む組成物の摂取により、コラーゲンペプチド特有の好ましくない食味を抑止できる。
本発明における組成物には、上記有効成分のほか、公知の添加物を配合することができる。添加物は特に限定されないが、経口摂取するために通常利用されるものが好ましく、例えば、賦形剤、結合剤、崩壊剤、潤沢剤、防腐剤、矯味剤、矯臭剤、着色剤、香料等の添加物を使用でき、皮膚改善作用が既知の素材を用いることができる。
本発明の組成物は、経口組成物であることが好ましい。経口組成物には、特に限定されないが、例えば、飲食品、医薬品、医薬部外品等が含まれ、飲食品であることが好ましく、飲料であることがより好ましい。飲食品としては、健康食品、栄養補助食品、特定保健用食品、サプリメント、乳製品、清涼飲料、用時溶解用粉末等が挙げられ、ペット用飲食品、家畜飼料等も含まれる。特に、用時溶解用粉末であることが好ましい。用時溶解用粉末は、さらにコーヒー、紅茶、ジュース、ヨーグルト、スープなどに溶解させたり、料理に混ぜて摂取することができるものである。用事溶解用粉末は、保存中の安定性や、好みのものに溶解又は添加して摂取できるという服用のしやすさの点で特に優れている。このほか、錠剤やカプセル剤の形態で用いることができる。医薬品、医薬部外品の形態は、典型的には、顆粒剤、錠剤、カプセル剤、液剤等の経口用製剤である。
本発明における組成物は、皮膚症状の予防・改善のために用いることができ、例えば、皮膚の保湿機能や弾力性の低下、肌の張りや艶の減少、荒れ、シワ、くすみ等の症状を予防または改善することができる。
本発明における組成物の製造は常法に従って行うことができるが、有効成分であるコラーゲンペプチド、エラスチンペプチドおよびプロテオグリカンはそのまま配合してよく、あるいは固体状、ペースト状または液体の形態で配合してもよい。また、必要に応じて皮膚改善作用を備える公知の成分や添加物等のその他の成分を配合する。有効成分やその他の成分の含有量や重量比は、上記した通りである。
本発明のさらなる態様としては、コラーゲンペプチド、エラスチンペプチドおよびプロテオグリカンを所定の比率で含む皮膚改善剤が挙げられる。皮膚改善剤中のコラーゲンペプチドの重量とエラスチンペプチドおよびプロテオグリカンの合計重量との重量比[(コラーゲンペプチド):(エラスチンペプチド+プロテオグリカン)]は、例えば、1:10~1:0.0000001であり、好ましくは1:5~1:0.000001、より好ましくは1:2~1:0.00001である。また、コラーゲンペプチドの重量とエラスチンペプチドの重量との重量比[(コラーゲンペプチド):(エラスチンペプチド)]は、例えば、1:5~1:0.000001であり、好ましくは1:2~1:0.00001、より好ましくは1:1~1:0.0001である。さらに、コラーゲンペプチドの重量とプロテオグリカンの重量との重量比[(コラーゲンペプチド):(プロテオグリカン)]は、例えば、1:0.2~1:0.0000001であり、好ましくは1:0.1~1:0.0000005、より好ましくは1:0.02~1:0.000002である。皮膚改善剤中に含まれる有効成分、その他の成分や、皮膚改善剤の形態および用量、作用・効果、製造方法は、組成物に関して記載した内容を適用することができる。
本発明の線維芽細胞増殖促進剤が皮膚線維芽細胞の増殖に及ぼす影響を以下の方法で調べた。
コラーゲンペプチド、エラスチンペプチドおよびプロテオグリカンを秤量し、滅菌水に溶解させ、孔径0.2 μmのメンブランフィルターでろ過滅菌後、下記表1に示す試料濃度となるよう、0.1%FBS含有DMEM培地で100倍希釈して試料2~6を調製した。対照として、滅菌水を0.1%FBS含有DMEM培地で100倍希釈した試料1を用いた。なお、各試験試料の濃度は、成分としての濃度で算出した。使用した原料の詳細を以下に示す。
コラーゲンペプチド:新田ゼラチン(株)製、分子量100-1800、純度100%
エラスチンペプチド:「美弾エラスチンFI」(日本水産(株)製)、純度100%
プロテオグリカン:「プロテオグリカンF」(一丸ファルコス(株)製)、分子量:約450,000、純度20%
成人由来正常ヒト皮膚線維芽細胞NHDF株(クラボウ(株)より購入)を10%ウシ血清(FBS)含有ダルベッコ変性MEM培地(DMEM、ニッスイ社製)中に接種し、96ウェルプレート中に10,000細胞/ウェルとなるように播種し、37℃、5%CO2の条件下で一晩培養した。試験試料は水に溶解後、上述の通り0.1%FBS含有DMEM培地で100倍希釈して細胞に添加した。各試験試料(200μl)を添加して24時間後に細胞生存率を測定した。細胞生存率はMTS試薬(Promega社製)を用いて細胞内脱水素酵素により還元され生成したホルマザン量から算出した。各試験試料の細胞生存率は1%の水を含む0.1%FBS含有DMEM培地のみを添加した試料の細胞生存率を100%とした場合の値(%)として求めた。ホルマザンは490nmにおける紫外吸収により測定した。なお、本発明者は、あらかじめ細胞が増殖も減少もしないFBS濃度を検討し、0.1%FBS含有DMEM培地で培養したときこの条件を満たすことを確認し、試験培地の血清濃度を0.1%に調製することで、試験試料のみの線維芽細胞増殖促進作用を定量的に評価できることにした。
評価結果を表1に示す。表1のデータから、コラーゲンペプチドにエラスチンペプチドを任意の割合で組み合わせると、コラーゲン単体よりも線維芽細胞の増殖促進効果を高めたが、コラーゲンペプチドおよびエラスチンペプチドにさらにプロテオグリカンを組み合わせた場合に、さらに増殖促進効果を高めることを確認した。また、試料5、6の結果から、コラーゲンペプチドに対するプロテオグリカンの配合比が非常に低い場合であっても、繊維芽細胞を顕著に増加させることが判明した。
NHDF株を10%FBS含有DMEM倍地中に摂取し、24ウェルプレート中に125,000細胞/ウェルとなるように播種し、37℃、5%CO2の条件下で一晩培養した。試験試料は水に溶解後、0.1%FBS含有DMEM培地で100倍希釈して細胞に添加した。各試験試料を添加して72時間後にコラーゲン産生量を測定した。コラーゲン産生量はCollagen Type I, Human, ELISA Kit,エーセル社製)を用いて細胞培養上清中に分泌されたI型コラーゲンを定量した。I型アテロコラーゲンに特異的なポリクローナル抗体を用いて発色させ、450nmにおける紫外吸収により測定した。各試験試料のコラーゲン産生量は、1%の水を含む0.1%FBS含有DMEM培地のみを添加した試料のコラーゲン産生量を100%とした場合の値(%)として求めた。細胞を培養する際、実施例1と同様に、培養液中の血清濃度を低くすることにより、コラーゲン合成への影響を低く抑えた。
評価結果を表2に示す。表2のデータから、コラーゲンペプチドにエラスチンペプチドおよびプロテオグリカンを組み合わせた場合に、コラーゲン単体よりも更にコラーゲンの産生促進効果を高めることを確認した。また、最も低い濃度のエラスチンペプチドおよびプロテオグリカンを含有する試料3のコラーゲン産生量が最も多かったことから、エラスチンペプチドおよびプロテオグリカンの濃度が低いほどコラーゲンの産生促進作用が強いことを見出した。
<試験試料の調製>
実施例1の方法に従って、下記表3に示す試料1~4を調製した。
NHDF株を、10%FBS含有DMEM培地中に接種し、96ウェルプレート中に10,000細胞/ウェルとなるように播種し、37℃、5%CO2の条件下で一晩培養した。試験試料は水に溶解後、0.1%FBS含有DMEM培地で100倍希釈して細胞に添加した。各試験試料を添加して24時間後にI型コラーゲン合成遺伝子(Col1a1)の発現を定量リアルタイムPCRにより定量した。コントロールのハウスキーピング遺伝子としてβ-actinを用いた。細胞をPBSで洗浄し、TaqMan Gene Expression Cell-to-CT Kit (Ambion社)のプロトコールに従いcDNAを作成した。このcDNAを鋳型として、TaqMan Fast Univ. Gene Expression Master Mix (Applied Biosystems社)を用いて、リアルタイムPCRを実施した。Real time PCRは、StepOnePlus Real Time PCR System (Applied Biosystems社)を用いた。解析は、ΔΔCT法を用い、試験試料無添加培養細胞の補正値を1とした比で表した。
評価結果を表3に示す。表3のデータから、コラーゲンペプチドにエラスチンペプチドおよびプロテオグリカンを組み合わせた場合に、コラーゲン単体よりもコラーゲン合成遺伝子の発現を高めることを確認した。
下記表4に示す配合量のコラーゲン、エラスチン、プロテオグリカン、デキストリンを秤量し、均一になるよう混合することで用時溶解用粉末(6.5 g)を製造した。得られた粉末を水、果汁、またはコンソメスープに溶解したところ、いずれも分散性はよく、用時溶解飲料として適切であった。
下記成分からなるソフトカプセル剤皮の中に、上記の用事溶解用粉末を常法により充填し、ソフトカプセルを得た。
ゼラチン 60.0%
グリセリン 30.0%
パラオキシ安息香酸メチル 0.15%
パラオキシ安息香酸プロピル 0.51%
水 適量
下記に示す配合の混合物を、常法により造粒ならびに成型し、錠剤を得た。
Claims (9)
- コラーゲンペプチド、エラスチンペプチドおよびプロテオグリカンを含む組成物であって、コラーゲンペプチドの重量とエラスチンペプチドおよびプロテオグリカンの合計重量との重量比[(コラーゲンペプチド):(エラスチンペプチド+プロテオグリカン)]が、1:10~1:0.0000001である、上記組成物。
- コラーゲンペプチドの重量とエラスチンペプチドの重量との重量比[(コラーゲンペプチド):(エラスチンペプチド)]が、1:5~1:0.000001である、請求項1に記載の組成物。
- コラーゲンペプチドの重量とプロテオグリカンの重量との重量比[(コラーゲンペプチド):(プロテオグリカン)]が、1:0.2~1:0.0000001である、請求項1に記載の組成物。
- 経口用である、請求項1~3のいずれか1項に記載の組成物。
- 飲食品である請求項1~3のいずれか1項に記載の組成物。
- コラーゲンペプチド、エラスチンペプチドおよびプロテオグリカンを含む皮膚改善剤であって、コラーゲンペプチドの重量とエラスチンペプチドおよびプロテオグリカンの合計重量との重量比[(コラーゲンペプチド):(エラスチンペプチド+プロテオグリカン)]が、1:10~1:0.0000001である、皮膚改善剤。
- 線維芽細胞増殖促進作用を備える、請求項6に記載の皮膚改善剤。
- コラーゲン産生促進作用を備える、請求項6または7に記載の皮膚改善剤。
- 皮膚老化防止作用を備える、請求項6~8のいずれか1項に記載の皮膚改善剤。
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CN201480066243.3A CN105792837B (zh) | 2013-12-04 | 2014-11-28 | 包含胶原蛋白肽、弹性蛋白肽及蛋白多糖的组合物 |
JP2015526437A JP5894341B2 (ja) | 2013-12-04 | 2014-11-28 | コラーゲンペプチド、エラスチンペプチドおよびプロテオグリカンを含む組成物 |
US15/100,785 US20160303021A1 (en) | 2013-12-04 | 2014-11-28 | Composition comprising collagen peptide, elastin peptide and proteoglycan |
HK16112667.9A HK1224223A1 (zh) | 2013-12-04 | 2016-11-03 | 包含膠原蛋白肽、彈性蛋白肽及蛋白多糖的組合物 |
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CN113115884A (zh) * | 2019-12-31 | 2021-07-16 | 中粮集团有限公司 | 复合肽固体饮料及其制备方法和应用 |
WO2024097704A1 (en) * | 2022-11-02 | 2024-05-10 | Zanda, Llc | Herbal formulation for improvement of skin growth |
KR20240107865A (ko) * | 2022-12-30 | 2024-07-09 | 충남대학교산학협력단 | 히알루론산, 엘라스틴 및 콜라겐을 함유하는 기능성 커피 및 이의 제조방법 |
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TW201609175A (zh) | 2016-03-16 |
JPWO2015083630A1 (ja) | 2017-03-16 |
US20160303021A1 (en) | 2016-10-20 |
TWI659746B (zh) | 2019-05-21 |
CN105792837B (zh) | 2020-08-25 |
HK1224223A1 (zh) | 2017-08-18 |
CN105792837A (zh) | 2016-07-20 |
JP5894341B2 (ja) | 2016-03-30 |
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