Elastin production enhancer and method for producing same

Abstract

To provide a novel application of components extracted from an ovary membrane of salmon.SOLUTION: There is provided an elastin production promoter containing a decomposed product by proteinase of an ovary membrane of salmon as an effective component. Further, there is provided a method for producing an elastin production promoter in which at least one kind selected from aroase (registered trademark, produced by Yakult Pharmaceutical Industry Co., Ltd.), papain, pancidase (registered trademark, produced by Yakult Pharmaceutical Industry Co., Ltd.) is added to the ovary membrane of salmon to perform enzymolysis, and a decomposed product by the enzymolysis is collected. It is preferable that the proteinase use aroase, papain and pancidase in combination.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an elastin production enhancer made from salmon ovarian membrane as a raw material, and a method for producing the same. [Background technology]

[0002] Conventionally, a rough skin improving agent containing a component extracted by treating salmon ovarian membrane with a protease has been disclosed in Patent Document 1.

[0003] Furthermore, Patent Document 2 discloses an agent for increasing IGF-1 levels that contains a component extracted by treating salmon ovarian membrane with a protease.

[0004] Furthermore, Patent Document 3 discloses an anti-aging agent that contains a component extracted by treating salmon ovarian membrane with a protease, and pharmacologically active components that can be added to foods and contain collagen, nucleic acid, elastin, hyaluronic acid, ceramide, and galactooligosaccharide. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Patent No. 3899116 [Patent Document 2] Patent No. 3946238 [Patent Document 3] Patent No. 6085137 Summary of the Invention [Problem to be solved by the invention]

[0006] However, there is hope for the development of many more applications for the ingredients extracted from salmon ovarian membrane.

[0007] The present invention has been made in consideration of the above-mentioned points, and has an object to provide a new use of components extracted from salmon ovarian membrane. [Means for solving the problem]

[0008] In order to achieve the above-mentioned object, one aspect of the present invention provides an elastin production promoter that is characterized by containing, as an active ingredient, a proteolytic enzyme digestion product of salmon ovarian membrane.

[0009] The elastin production-enhancing agent of the present invention can be orally ingested to promote the production of elastin in the body of a person who has orally ingested the elastin production-enhancing agent, and as a result, various physiologically active effects due to elastin can be expected.

[0010] In the elastin production-enhancing agent of the present invention, the protease is preferably a neutral protease.

[0011] Moreover, the protease is more preferably at least one selected from the group consisting of alloase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.), papain, and punchidase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.).

[0012] Furthermore, the protease is more preferably alloase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.), papain, or punchidase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.).

[0013] In addition, it is preferable that the elastin production promoter of the present invention is used for at least one application selected from the following: regeneration of damaged ligaments, protection of the epidermis against ultraviolet rays, suppression of vascular endothelial damage, inhibition of platelet aggregation, antithrombosis, improvement of blood flow, and suppression of breast sagging.

[0014] Another aspect of the present invention provides a method for producing an elastin production promoter, which comprises adding at least one enzyme selected from the group consisting of alloase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.), papain, and punchidase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.) to salmon ovarian membrane to effect enzymatic hydrolysis, and collecting a product of the enzymatic hydrolysis.

[0015] According to the method for producing an elastin production enhancer of the present invention, an elastin production enhancer having excellent elastin production-enhancing effects can be obtained with good productivity.

[0016] In the method for producing the elastin production-enhancing agent of the present invention, it is preferable to carry out the enzymatic degradation using a combination of Alloase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.), papain, and Panchidase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.).

[0017] Furthermore, from another aspect, the present invention provides a method for promoting elastin production, in which a proteolytic enzyme-derived hydrolyzate of salmon ovarian membrane is orally ingested to promote the production of elastin in the body of a person who orally ingests the product.

[0018] According to this method for promoting elastin production, by orally ingesting the protease-degraded product of salmon ovarian membrane according to the present invention as an active ingredient, it is possible to promote the production of elastin in the body of a person who orally ingests the elastin production promoter, and as a result, various physiological activity effects due to elastin can be expected.

[0019] In this method, the proteolytic enzyme is preferably a neutral protease.

[0020] Furthermore, the protease is preferably at least one selected from the group consisting of alloase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.), papain, and punchidase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.).

[0021] Here, the protease may be Alloase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.), papain, or Panchidase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.).

[0022] Furthermore, this method for promoting elastin production can be used for at least one application selected from the following: regeneration of damaged ligaments, protection of the epidermis against ultraviolet rays, suppression of vascular endothelial damage, inhibition of platelet aggregation, antithrombosis, improvement of blood flow, and suppression of breast sagging. Effect of the Invention

[0023] According to the present invention, it is possible to provide an elastin production-promoting agent that promotes the production of elastin in a person who takes it. [Brief description of the drawings]

[0024] [Figure 1] 1 is a flowchart showing a method for producing an elastin production-enhancing agent. [Diagram 2] 1 is a graph showing changes in plasma concentration of Pro-Gly, a type of elastin peptide, in an example. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] Hereinafter, one embodiment of the elastin production-enhancing agent according to the present invention will be described with reference to the drawings.

[0026] The elastin production-enhancing agent of the present invention contains a component extracted by treating the ovarian membrane of salmon (order Salmoniformes, family Salmonidae) with a protease (also referred to as a salmon ovarian membrane extract component).

[0027] As the salmon ovarian membrane, for example, a membranous muscle (ovarian skin) remaining when salmon roe is processed into salmon roe can be used. It is preferable to use raw salmon ovarian membrane that has not been processed by heating, drying, or the like.

[0028] As the proteolytic enzyme, one or more enzymes selected from various proteases can be used. Preferably, neutral proteases are used. Among them, Aroase (registered trademark), Papain, and Panchidase (registered trademark) are preferably used, and it is most preferable to use these three enzymes in combination.

[0029] Papain is a type of proteolytic enzyme extracted from the latex of papaya fruit, and is used for protein analysis and as a meat tenderizer, etc. It works well around neutral pH, but the optimum pH is 6.5 to 7.0.

[0030] Aroase (registered trademark) is a neutral protease sold by Yakult Pharmaceutical Co., Ltd. As Aroase (registered trademark), for example, "Aroase NP-10" (trade name, manufactured by Yakult Pharmaceutical Co., Ltd.) can be used.

[0031] Panchidase (registered trademark) is another neutral protease sold by Yakult Pharmaceutical Co., Ltd. As Panchidase (registered trademark), for example, "Panchidase NP-2" (trade name, manufactured by Yakult Pharmaceutical Co., Ltd.) can be used.

[0032] The salmon ovarian membrane extract can be obtained, for example, by the method shown in Fig. 1. That is, the ovarian membrane separated from salmon eggs is treated with a protease to extract a solution, that is, the product of the ovarian membrane decomposition by the enzyme is collected (step S01). The solution obtained in step S01 is filtered (step S02). The filtrate obtained in step S02 is dried (step S03).

[0033] Specifically, in step S01, the raw material salmon ovarian membrane is first thoroughly washed with water, and then water is added. For example, water may be added so that the mass ratio of raw ovarian membrane:water is 1:1 to 5.

[0034] Next, the mixed solution is heated to a temperature suitable for the enzymatic reaction of the protease, preferably to 40 to 60° C., and then the protease is added. For example, the protease is added in an amount of preferably 0.1 to 5.0 parts by weight, more preferably 0.2 to 2.0 parts by weight, based on 100 parts by weight of the total amount of the ovarian membrane.

[0035] When Aroase (registered trademark), papain, or punchidase (registered trademark) is used as the protease, Aroase (registered trademark) is preferably added at 0.05 to 0.50 parts by mass, punchidase (registered trademark) is preferably added at 0.10 to 1.00 parts by mass, and papain is preferably added at 0.05 to 0.50 parts by mass, relative to 100 parts by weight of the total amount of raw ovarian membrane. The mass ratio of Aroase (registered trademark), papain, and punchidase (registered trademark) is preferably 1:1:1 to 1:1:4.

[0036] Then, the mixture is subjected to an enzymatic decomposition reaction with the protease. When the protease contains, for example, Aroase (registered trademark), Papain, or Panchidase (registered trademark), this enzymatic decomposition reaction is preferably carried out for 2 to 6 hours at a temperature range of 45 to 50° C. The ovarian membrane is decomposed by this enzymatic decomposition reaction.

[0037] Next, the protease contained in the suspension obtained by the enzymatic decomposition reaction is inactivated. The protease can be inactivated, for example, by increasing the temperature of the suspension and maintaining the increased temperature. For example, the temperature of the suspension can be increased to 95°C and maintained at this temperature for about 30 minutes.

[0038] Activated carbon is added to the suspension in which the protease has been inactivated, and the suspension is deodorized, decolorized, and degreased. Activated carbon is preferably added in an amount of, for example, about 1 to 10% by mass relative to the raw ovarian membrane, and after the activated carbon is added, the suspension is preferably stirred for 1 to 3 hours.

[0039] In step S02, the suspension to which the activated carbon has been added is centrifuged. For example, a decanter may be used to centrifuge the suspension. Next, the supernatant obtained by the centrifugation is filtered through diatomaceous earth to obtain an extract from which solids in the suspension have been removed.

[0040] In step S03, the extract obtained in step S02 is concentrated. For example, a centrifugal thin film evaporator may be used to concentrate the extract. For example, a CONTROL may be used as the centrifugal thin film evaporator. The concentrated extract is preferably subjected to heat sterilization. Heat sterilization is preferably performed at 60 to 100°C for 20 to 60 minutes.

[0041] The heat-sterilized extract is dried. The method for drying the extract is not particularly limited, and can be, for example, spray drying, vacuum drying, freeze drying, drum drying, hot air drying, etc. The powder obtained by drying can be mixed with an excipient as necessary. For example, dextrin, starch, etc. can be mentioned. In addition, the powder may be passed through a sieve of, for example, 16 to 250 mesh so that the powder has a desired particle size or less. In this way, by passing the powder through a sieve, a powder of a predetermined particle size can be obtained, and for example, unintended substances mixed in the powder can be removed.

[0042] In addition, the elastin production promoter may contain other components in addition to the salmon ovarian membrane extract component, as long as the effects of the present invention are not inhibited. Examples of such components include ceramide, plasmalogen, krill oil, α-lipoic acid, coenzyme Q10, astaxanthin, β-carotene, fucoxanthin, hyaluronic acid, fucoidan, proteoglycan, glucosamine, collagen, placenta, liver powder, L-carnitine, ornithine, GABA, theanine, cystine, polyamine, royal jelly, soy isoflavone, resveratrol, hesperidin, melon extract, black pepper extract, long pepper extract, ginseng extract, artichoke leaf extract, amla extract, dokudami extract, ginger extract, cherry blossom extract, chamomile extract, lemon balm extract, echinacea extract, mimosa bark extract, vitamin B group, vitamin C, etc.

[0043] The form of the elastin production-enhancing agent of the present invention is not particularly limited, and may be in various forms such as tablets, chewable tablets, powders, capsules, granules, drinks, gels, syrups, liquid foods for tube-enteral nutrition, etc. The agent may also be in the form of foods and drinks such as health foods, functional foods, foods and drinks for specified health uses, foods and drinks for the sick, food additives therefor, pharmaceuticals, supplements, animal feed for livestock, racehorses, ornamental animals, pets, etc. The content of the salmon ovarian membrane extract in these products is preferably 0.33 to 100% by mass, more preferably 0.50 to 100% by mass.

[0044] The dosage of the elastin production-enhancing agent according to the present invention can be appropriately changed depending on the species, age, sex, etc. of the individual to which it is applied. For example, the dosage for humans is preferably an amount that allows an adult to ingest 0.1 to 10 g of salmon ovarian membrane extract per day, more preferably an amount that allows an adult to ingest 0.2 to 1.0 g. The daily dosage may be administered once, or may be administered in several divided doses. It is more preferable that the agent is administered orally. The subject of application is not limited to humans, and the agent can also be applied to animals such as dogs and cats.

[0045] The elastin production promoter of the present invention has the effect of promoting the production of elastin in the body of the person who takes it. Specifically, it has been confirmed that when the person takes the elastin promoter, the blood concentration of Pro-Gly, which is a type of elastin peptide, increases in the blood of the person who takes it. Pro-Gly is a dipeptide composed of proline and glycine, and is known to promote the production of elastin.

[0046] Elastin has been reported to be useful for: (1) regenerating damaged ligaments; (2) protecting the epidermis against ultraviolet rays; (3) suppressing damage to the vascular endothelium; (4) inhibiting platelet aggregation, having antithrombotic effects, and improving blood flow; and (5) preventing breast sagging.

[0047] Specifically, (1) regeneration of damaged ligaments is described, for example, in "Development of Tissue-Engineered Ligaments Elastin Promotes Regeneration of the Rabbit Medial Collateral Ligament" (Artif. Organs, 42(6):E102-E113, 2017). According to this literature, when rabbits with ruptured medial collateral ligaments were injected with porcine elastin for six weeks, collagen and elastin gene expression was confirmed, the protein content in the cells increased, and the elastic modulus of the medial collateral ligament also increased. This suggests that elastin can heal ligaments damaged by sports activities or injuries.

[0048] (2) The epidermis protective effect against UV rays is described, for example, in "Protective Effect of Bovine Elastin Peptides against Photoaging in Mice and Identification of Novel Antiphotoaging Peptides" (J. Agric. Food Chem., 66 (41), 2018). According to this document, when bovine elastin hydrolysate was orally administered to mice with UV-damaged skin, epidermal hyperplasia and apoptosis of fibroblasts were suppressed. Furthermore, it was revealed that mice orally administered the elastin hydrolysate had elastase inhibitory activity. This suggests that elastin hydrolysate can protect UV-damaged skin.

[0049] (3) The inhibition of vascular endothelial damage is described, for example, in "Prophylactic effects of elastin peptide derived from the bulbus arteriosus of fish on vascular dysfunction in spontaneously hypertensive rats" (Life Sciences 120, pp.48-53, 2014). According to this document, when spontaneously hypertensive rats were fed bonito-derived elastin peptide for five weeks, damage to the aortic endothelium was inhibited compared to a group that did not receive elastin peptide. This suggests that elastin peptides can prevent vascular dysfunction associated with hypertension.

[0050] (4) Inhibition of platelet aggregation, antithrombotic effects, and improvement of blood flow are described, for example, in "Elastin peptides prepared from piscine and mammalian elastic tissues inhibit and mammalian elastic tissues inhibit collagen-induced platelet aggregation and stimulate migration and proliferation of human skin fibroblasts" (J. Pept. Sci., 16(11):652-8, 2010). According to this document, when elastin peptides derived from bovine ligament, porcine aorta, and bonito bulbus arteriosus were applied to anticoagulated blood, platelet aggregation was suppressed. From this, elastin peptides can be expected to prevent cerebral infarction and myocardial infarction caused by thrombus formation, and improve blood flow.

[0051] (5) The effect of suppressing breast sagging is described, for example, in "The effect of bonito-derived elastin peptides in suppressing breast sagging and improving breast skin elasticity during dieting in healthy Japanese women (Functional Foods and Pharmacological Nutrition, 3(10), pp.131-138, 2016)." According to this document, when humans ingested bonito arterial bulb-derived elastin peptides for eight weeks, the height from the floor to the top of the bust, the angle from the underbust to the top, and the elasticity of the skin of the bust increased significantly compared to before ingestion. From this, it can be expected that humans who ingest elastin peptides will be able to maintain a beautiful bust line that does not sag.

[0052] Therefore, the elastin production-enhancing agent of the present invention can be used for various applications as described above. EXAMPLES

[0053] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples in any way.

[0054] [Example] (Production of elastin production promoter consisting of salmon ovarian membrane extract) 200 parts by mass of water was added to 100 parts by mass of raw ovarian membrane separated from salmon eggs and thoroughly washed. 0.1 parts by mass of papain, 0.1 parts by mass of "Aroase NP-10" (trade name, manufactured by Yakult Pharmaceutical Co., Ltd.), and 0.4 parts by mass of "Panthidase NP-2" (trade name, manufactured by Yakult Pharmaceutical Co., Ltd.) were added to this mixture relative to 100 parts by mass of raw ovarian membrane used as a raw material, and enzymatic decomposition was carried out at a temperature of 45 to 50°C for 4 hours. Next, the temperature of the mixture was raised to 95°C and maintained for 30 minutes to inactivate the enzyme.

[0055] Next, 3 parts by mass of activated carbon was added to the mixture, and the mixture was deodorized, decolorized, and degreased. The mixture was then centrifuged to collect the supernatant, which was then filtered through diatomaceous earth to remove solids.

[0056] The filtrate was concentrated using a centrifugal thin-film evaporator (CONTROL), sterilized at 90°C for 30 minutes, and then spray-dried to obtain a powder. This powder was passed through a 16-mesh sieve to remove coarse particles, obtaining a salmon ovarian membrane extract. This salmon ovarian membrane extract was used as an elastin production promoter in the following test examples. The salmon ovarian membrane extract is sold by Nihon Shokukin Kogyo Co., Ltd. under the names SOP (Salmon Ovary Peptide) and Salmon Placenta (both registered trademarks).

[0057] [Test Example] (Single Ingestion Test of Elastin Production Enhancer) (1) Test method subject The information about the subjects is as follows. Subjects were selected if they met the following condition (b) and did not meet the condition (c). (a) Number of subjects: 2 (b) Subject selection criteria: Healthy adult men and women (c) Exclusion criteria for subjects: Subjects undergoing or requiring treatment for a disease, and those who are pregnant or breastfeeding.

[0058] (2) Test food The subjects were provided with 2 g of a powdered elastin production enhancer as a test food.

[0059] (3) Study design The test was carried out in the following manner. 3-1) A single intake test of the test food was conducted by two subjects. 3-2) The test was started at 9:00 a.m. on the day of ingestion under the condition that subjects had not eaten or drunk anything since 9:00 p.m. on the day before ingestion (tap water, mineral water, etc. were allowed). The subjects orally ingested the test food (wrapped in an oblate and ingested the test food). 3-3) Blood sampling: Blood samples were taken five times in total: before ingestion of the test food, 30 minutes after ingestion, 1 hour, 4 hours, and 6 hours after ingestion. 3-4) Blood collection amount: 5mL / time

[0060] (4) Evaluation method The test evaluation involved measuring the concentration of a dipeptide (elastin peptide Pro-Gly) in plasma using LC / MS (Liquid Chromatography-Mass spectrometry) and evaluating the change in concentration before and after intake.

[0061] Figure 2 shows the changes in the Pro-Gly concentration in the subjects' plasma. As shown in Figure 2, when the subjects orally took the elastin production enhancer, the Pro-Gly concentration in the blood reached its highest level 30 minutes later.

[0062] One hour after oral ingestion, the concentration of Pro-Gly in the blood decreased, i.e., the concentration of Pro-Gly in the blood one hour after oral ingestion was lower than the concentration 30 minutes after oral ingestion, but higher than the concentration before oral ingestion.

[0063] Four hours after oral ingestion, the concentration of Pro-Gly in the blood decreased slightly, i.e., the concentration of Pro-Gly in the blood four hours after oral ingestion was slightly lower than the concentration one hour after oral ingestion, but higher than the concentration before oral ingestion.

[0064] Six hours after oral ingestion, the concentration of Pro-Gly in the blood decreased slightly, i.e., the concentration of Pro-Gly in the blood 6 hours after oral ingestion was slightly lower than the concentration 4 hours after oral ingestion, but higher than the concentration before oral ingestion.

[0065] In this way, the elastin production-enhancing agent enhances the production of elastin in the person who takes it, and as a result, it becomes possible to increase the Pro-Gly concentration in the subject's blood.

Claims

1. An elastin production promoter comprising as an active ingredient a protease digest of salmon ovarian membrane, the protease digesting the membrane using alloase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.), papain, and punchidase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.).

2. The elastin production-promoting agent according to claim 1, which is used for at least one purpose selected from the following: regeneration of damaged ligaments, protection of the epidermis against ultraviolet rays, inhibition of vascular endothelial damage, inhibition of platelet aggregation, antithrombosis, improvement of blood flow, and inhibition of breast sagging.

3. A method for producing an elastin production promoter, comprising the steps of adding alloase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.), papain, and punchidase (registered trademark, manufactured by Yakult Pharmaceutical Co., Ltd.) to salmon ovarian membrane to effect enzymatic hydrolysis, collecting a hydrolyzate produced by the enzymatic hydrolysis, and incorporating the hydrolyzate as an active ingredient.

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