Oral composition containing nattokinase

Abstract

To provide an oral composition that contains nattokinase and oil solution and shows an increase in the initial activity or post-storage activity of nattokinase.SOLUTION: An oral composition contains nattokinase and oil solution in combination with an amino acid, so that the oral composition shows an increase in the initial activity or post-storage activity of nattokinase.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an oral composition comprising nattokinase and an oil agent, in which the activity of nattokinase immediately after production (initial activity) and the activity of nattokinase after storage (post-storage activity) are improved. [Background technology]

[0002] Thrombosis is a pathological condition in which blood clots form in blood vessels, obstructing blood flow in the circulatory system, and is known to be a factor in the development of serious diseases such as cerebral infarction, myocardial infarction, and pulmonary infarction. Conventionally, methods for preventing or treating thrombosis have been used, such as administering drugs such as antiplatelet agents, anticoagulants, and thrombolytic agents to prevent thrombus formation or dissolve thrombus. However, these drugs are accompanied by side effects and require administration under the supervision of a physician, and therefore cannot be taken easily and on a daily basis.

[0003] Meanwhile, natto, a traditional Japanese food, has been reassessed as a health food since it was reported to contain nattokinase, which has thrombolytic activity. However, natto has a distinctive odor and stickiness, and many people do not eat natto. Therefore, to facilitate the intake of nattokinase, nattokinase has been formulated into food products such as capsules and tablets. Such nattokinase-containing foods are useful for self-medication because they can be easily taken daily without the need for medical supervision.

[0004] Various techniques for formulating nattokinase into oral compositions have been reported. For example, Patent Document 1 reports that a food composition containing a freeze-dried milk fermentation product, nattokinase, and a food-safe carrier is easy to ingest and provides beneficial effects derived from the milk fermentation product and nattokinase. Patent Document 2 also reports that a granular soybean processed food with an average particle size of 100 to 500 μm, obtained by granulating a powder mixture containing 40 to 80% by mass of soybean powder, 0.5 to 15% by mass of nattokinase powder, and 3 to 30% by mass of oligosaccharides, has no natto odor and exhibits good dispersibility in liquid and creamy foods. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] WO 02 / 76240 [Patent Document 2] Japanese Patent Application Laid-Open No. 2005-130727 Summary of the Invention [Problem to be solved by the invention]

[0006] The present inventors have conducted research to develop an oral composition containing nattokinase and an oil agent, and have found that the oral composition has a problem in that the activity of nattokinase decreases after storage (post-storage activity). Furthermore, in developing the oral composition, it is also important to improve the activity of nattokinase immediately after production (initial activity).

[0007] Therefore, an object of the present invention is to provide an oral composition containing nattokinase and an oil agent, which has improved initial activity or activity after storage of nattokinase. [Means for solving the problem]

[0008] The present inventors have conducted extensive research to solve the above problems and have found that the initial activity of nattokinase and the activity after storage can be improved by adding an amino acid to nattokinase and an oil agent in an oral composition. Based on this finding and through further research, the present invention has been completed.

[0009] That is, the present invention provides the following aspects. Item 1. An oral composition comprising (A) nattokinase, (B) an amino acid, and (C) an oil solution. Item 2. The oral composition according to Item 1, wherein the component (B) is at least one selected from the group consisting of arginine, aspartic acid, phenylalanine, lysine, isoleucine, glutamine, methionine, tyrosine, tryptophan, histidine, glutamic acid, glycine, and proline. Item 3. The oral composition according to Item 1 or 2, wherein the component (B) is at least one selected from the group consisting of arginine, aspartic acid, phenylalanine, lysine, isoleucine, glutamine, methionine, tyrosine, tryptophan, histidine, and glutamic acid. Item 4. The oral composition according to any one of Items 1 to 3, which is a food product. Item 5. A method for improving nattokinase activity in an oral composition containing nattokinase and an oil agent, comprising: The method, wherein the oral composition contains (A) nattokinase, (B) an amino acid, and (C) an oil. Item 6. An activity enhancer containing an amino acid as an active ingredient, which is used to enhance the activity of nattokinase in an oral composition containing nattokinase and an oil agent. [Effects of the Invention]

[0010] According to the present invention, an oral composition containing nattokinase and an oil agent has improved initial activity or activity after storage, making it possible to provide an oral composition that can effectively exert the action of nattokinase. DETAILED DESCRIPTION OF THE INVENTION

[0011] 1. Oral Composition The oral composition of the present invention is characterized by containing (A) nattokinase, (B) an amino acid, and (C) an oil solution. The oral composition of the present invention is described in detail below.

[0012] [(A) Nattokinase] The oral composition of the present invention contains nattokinase (sometimes referred to as component (A)). Nattokinase is an enzyme that has a fibrinolytic activity and is produced by Bacillus subtilis var. natto.

[0013] The nattokinase used in the present invention can be obtained by a known production method. Specific production methods for nattokinase include a method of culturing Bacillus subtilis natto, a method of obtaining it from a transformant incorporating a gene encoding nattokinase, and a method of synthesizing it by chemical synthesis. The nattokinase used in the present invention may be obtained by any production method, but from the viewpoint of reducing production costs, etc., nattokinase obtained by a method of culturing Bacillus subtilis natto is preferred.

[0014] The nattokinase used in the present invention may be a purified product, or may be in an unpurified state as long as it is edible. For example, when using nattokinase obtained by culturing Bacillus subtilis natto, it may be an extract of the culture of Bacillus subtilis natto. Furthermore, it may be nattokinase obtained by subjecting a culture of Bacillus subtilis natto to ion exchange chromatography, gel filtration chromatography, hydrophobic chromatography, or the like to purify the nattokinase, or it may be nattokinase obtained by subjecting a culture of Bacillus subtilis natto to a crude purification process such as solid-liquid separation as necessary, followed by removal of water or drying.

[0015] Nattokinase is commercially available as a powder product containing excipients, a crude product, a purified product, etc., and these commercially available products can also be used as nattokinase in the present invention.

[0016] In the oral composition of the present invention, the content of component (A) may be appropriately determined depending on the oral form, etc., and, for example, the amount of nattokinase per 1 g of the oral composition of the present invention is 100 FU or more, preferably 300 to 15,000 FU, more preferably 500 to 5,000 FU, and particularly preferably 800 to 1,500 FU.

[0017] In the present invention, the "FU" indicating the activity of nattokinase is a fibrinolytic activity unit according to the standards for natto culture extract foods published by the Japan Health and Nutrition Food Association on January 15, 2003.

[0018] [(B) Amino acid] The oral composition of the present invention contains an amino acid (sometimes referred to as component (B)) in addition to nattokinase. In the oral composition of the present invention, the coexistence of nattokinase and an amino acid in the presence of an oil agent makes it possible to improve the initial activity or activity after storage of nattokinase.

[0019] The amino acids used as component (B) may be in the L-, D-, or DL-form, but are preferably in the L-form. Furthermore, of the amino acids used as component (B), aspartic acid and glutamic acid may be in the form of a salt such as a sodium salt.

[0020] Of the components (B), preferred examples include arginine, aspartic acid, phenylalanine, lysine, isoleucine, glutamine, methionine, tyrosine, tryptophan, histidine, glutamic acid, glycine, and proline.

[0021] Furthermore, among the (B) components, arginine, aspartic acid, phenylalanine, lysine, isoleucine, glutamine, methionine, tyrosine, tryptophan, histidine, and glutamic acid can improve both the initial activity and the activity after storage of nattokinase, and are therefore preferably used as the (B) component in the present invention. Among these amino acids, arginine, aspartic acid, phenylalanine, and lysine are preferred from the viewpoint of effectively improving both the initial activity and the activity after storage of nattokinase in a balanced manner. In particular, when arginine is used as the (B) component, nattokinase The difference between the initial activity of nattokinase and the activity after storage is small, making it possible to effectively suppress the decrease in nattokinase activity due to storage.

[0022] As component (B), one type of amino acid may be selected and used alone, or two or more types of amino acids may be used in combination.

[0023] The content of component (B) in the oral composition of the present invention may be appropriately set depending on the formulation form of the oral composition, etc., and may be, for example, 0.01 to 30% by weight, preferably 0.1 to 15% by weight, and more preferably 1 to 5% by weight.

[0024] In the oral composition of the present invention, the ratio of component (B) to component (A) is not particularly limited, but examples thereof include 0.01 to 5 g, preferably 0.01 to 1 g, and more preferably 0.01 to 0.5 g of component (B) per 10,000 FU of component (A).

[0025] [(C) Oil] The oral composition of the present invention contains an oil agent (sometimes referred to as component (C)).

[0026] The type of oil is not particularly limited as long as it is edible, and examples thereof include vegetable oils, animal oils, waxes, higher fatty acids, higher alcohols, cholesterol, and the like.

[0027] Specific examples of vegetable oils include soybean oil, macadamia nut oil, olive oil, rice germ oil, safflower oil, cottonseed oil, palm oil, almond oil, avocado oil, camellia oil, persic oil, sesame oil, corn oil, castor oil, jojoba oil, carnauba wax, candelilla wax, cacao butter, kukui nut oil, shea butter, evening primrose oil, perilla oil, tea seed oil, rapeseed oil, palm kernel oil, palm oil, peanut oil, sunflower oil, grape seed oil, meadowhoo oil, and hydrogenated oils thereof.

[0028] Specific examples of animal oils include egg yolk oil, lanolin, beef tallow, lard, horse oil, mink oil, fish oil, and hardened oils thereof.

[0029] Specific examples of wax include beeswax, carnauba wax, and candelilla wax.

[0030] Examples of higher fatty acids include fatty acids having 8 to 30 carbon atoms, preferably 10 to 20 carbon atoms, and specific examples include lauric acid, myristic acid, palmitic acid, sebacic acid, oleic acid, stearic acid, linoleic acid, etc. These fatty acids may be used alone or in combination of two or more.

[0031] Examples of higher alcohols include alcohols having preferably 12 to 22 carbon atoms, and specific examples include lauryl alcohol, myristyl alcohol, palmityl alcohol, cetanol, oleyl alcohol, stearyl alcohol, isostearyl alcohol, cetostearyl alcohol, and behenyl alcohol. Examples of the fatty acids include those having 8 to 30 carbon atoms, preferably 10 to 20 carbon atoms, and specific examples thereof include lauric acid, myristic acid, palmitic acid, sebacic acid, oleic acid, stearic acid, and linoleic acid.

[0032] These components (C) may be used alone or in combination of two or more.

[0033] Of the components (C), vegetable oils and waxes are preferred.

[0034] The content of component (C) in the oral composition of the present invention may be appropriately set depending on the formulation form of the oral composition, etc., and may be, for example, 5 to 95% by weight, preferably 40 to 95% by weight, and more preferably 60 to 90% by weight.

[0035] [(D) Surfactant] In addition to the components described above, the oral composition of the present invention may contain a surfactant (sometimes referred to as component (D)) as needed.

[0036] The type of oil is not particularly limited as long as it is edible, and any of nonionic surfactants, anionic surfactants, cationic surfactants, and zwitterionic surfactants may be used, with nonionic surfactants being preferred.

[0037] Examples of nonionic surfactants include glycerin fatty acid esters, lecithin, lecithin derivatives, polyoxyethylene hydrogenated castor oil, polyoxyethylene alkyl ethers, polyglycerin fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene glycerin fatty acid esters, sorbitan fatty acid esters, polyoxyethylene sorbit fatty acid esters, and polyethylene glycol fatty acid esters.

[0038] Of the components (D), preferred are glycerin fatty acid esters, lecithin and derivatives thereof.

[0039] Examples of glycerin fatty acid esters include esters of glycerin and fatty acids having 10 to 20 carbon atoms. Examples of fatty acids constituting glycerin fatty acid esters include fatty acids having 10 to 20 carbon atoms, such as capric acid, lauric acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, and arachidonic acid. Among these fatty acids, fatty acids having 12 to 20 carbon atoms are preferred, and fatty acids having 14 to 20 carbon atoms are more preferred. Furthermore, the glycerin fatty acid ester may be any of monoesters, diesters, and triesters, with monoesters being preferred.

[0040] When the oral composition of the present invention contains component (D), its content may be appropriately determined depending on the formulation of the oral composition, etc., and may be, for example, 0.01 to 10% by weight, preferably 0.05 to 10% by weight, and more preferably 0.1 to 8% by weight.

[0041] [Other ingredients] The oral composition of the present invention may contain other nutritional components or pharmacological components in addition to the above-mentioned components. Such nutritional components and pharmacological components are not particularly limited as long as they are usable in foods and oral pharmaceuticals, and examples thereof include vitamins, amino acids other than those mentioned above, minerals, carbohydrates, fatty acids, flavorings, seasonings, plant extracts, antioxidants, hypoglycemic agents, anticholesterol agents, immunostimulants, etc. These components may be used alone or in combination of two or more. The content of these components is appropriately determined depending on the type of components used and the intended use of the oral composition.

[0042] Furthermore, in order to prepare the oral composition of the present invention into a desired formulation, it may contain, as necessary, bases, additives, etc. in addition to the above-mentioned components. Such bases and additives are not particularly limited as long as they are usable in foods and pharmaceuticals, and examples thereof include water, water-soluble polymers, lower alcohols, polyhydric alcohols, pH adjusters, buffers, antioxidants, preservatives, thickeners, chelating agents, etc. These may be used alone or in combination of two or more. The content of these bases and additives is appropriately determined depending on the types of components used and the intended use of the oral composition.

[0043] [Dosage form / preparation type] The dosage form of the oral composition of the present invention is not particularly limited as long as it can be orally ingested or administered, and may be any of solid, semi-solid, or liquid, and may be appropriately selected depending on the type and use of the oral composition.

[0044] The formulation of the oral composition of the present invention is not particularly limited as long as it can be orally ingested or administered, and specific examples include foods and oral medicines.

[0045] When the oral composition of the present invention is formulated into a food product, the aforementioned components may be prepared as desired, either directly or in combination with other food materials or additives. Examples of such foods and beverages include general foods and beverages, as well as foods for specified health uses, foods with nutrient function claims, foods with functional claims, and foods for patients. The form of these foods is not particularly limited, but specific examples include supplements such as capsules (soft capsules, hard capsules), tablets, granules, powders, and jellies; beverages such as energy drinks, fruit juice drinks, and carbonated drinks; and luxury items such as dumplings, ice cream, sherbet, gummies, and candies. Among these foods and beverages, supplements are preferred, capsules, tablets, and granules are more preferred, and capsules are even more preferred.

[0046] When the oral composition of the present invention is formulated into an oral pharmaceutical preparation, the aforementioned components may be prepared into a desired form either as is or in combination with other additive components. Specific examples of such oral pharmaceutical preparations include capsules (soft capsules, hard capsules), tablets, granules, powders, jellies, syrups, etc. Among these oral pharmaceutical preparations, capsules, tablets, granules, and powders are preferred, and capsules are more preferred.

[0047] [Manufacturing method] There are no particular limitations on the method for producing the oral composition of the present invention. In one preferred embodiment, component (C) is mixed under heating at 50°C or higher, preferably 50 to 80°C, and then allowed to cool to 35°C or lower, and either components (B) and (A) are added simultaneously, or component (C) is added after component (B) has been added; in another more preferred embodiment, component (C) is mixed under heating at 50°C or higher, preferably 50 to 80°C, and then allowed to cool to 31 to 35°C, at which point component (B) is added, and component (A) is then added when the temperature has cooled to 30°C or lower.

[0048] 2. Methods for improving nattokinase activity and nattokinase activity enhancers The method for improving activity of the present invention is a method for improving the activity of nattokinase in an oral composition containing nattokinase and an oil agent, and is characterized in that the oral composition contains (A) nattokinase, (B) at least one amino acid selected from the group consisting of arginine, aspartic acid, phenylalanine, lysine, isoleucine, glutamine, methionine, tyrosine, tryptophan, histidine, and glutamic acid, and (C) an oil agent.

[0049] Furthermore, the activity enhancer of the present invention is used to improve the activity of nattokinase in an oral composition containing nattokinase and an oil agent, and is characterized in that it contains, as an active ingredient, (B) at least one amino acid selected from the group consisting of arginine, aspartic acid, phenylalanine, lysine, isoleucine, glutamine, methionine, tyrosine, tryptophan, histidine, and glutamic acid.

[0050] The activity improving method and activity improver of the present invention make it possible to improve the initial activity or post-storage activity of nattokinase in an oral composition containing nattokinase and an oil agent. The types and amounts of ingredients used in the activity improving method and activity improver, as well as specific embodiments, are as described in the section "1. Oral Composition" above. [Example]

[0051] The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples.

[0052] Test Example 1 An oral composition was prepared having the composition shown in Table 1. Specifically, predetermined amounts of soybean oil, beeswax, and glycerin fatty acid ester were mixed and dissolved at 70°C, and then a predetermined amount of lecithin was added and mixed. The mixture was then allowed to cool while stirring, and when the temperature reached 35°C, a predetermined amount of amino acid was added and mixed. When the temperature reached 30°C, nattokinase powder was added and mixed for 10 minutes to prepare an oral composition.

[0053] 50 g of each of the obtained oral compositions was filled into a glass bottle and left to stand for 2 weeks at 35°C under light-shielded conditions. For each oral composition, the nattokinase activity immediately after preparation (initial activity) and after standing for 2 weeks (activity after storage) were measured by the following method.

[0054] <Method for measuring nattokinase activity> A fibrinogen solution was prepared by dissolving fibrinogen ("Fibrinogen Sigma F8630", Sigma-Aldrich) in 0.17 M borate buffer to a concentration of 0.375 wt %. Separately, a thrombin solution was prepared by dissolving thrombin ("Thrombin Sigma T6634", Sigma-Aldrich) in 0.17 M borate buffer to a concentration of 100 U / ml. Next, 10 ml of the fibrinogen solution and 20 μl of the thrombin solution were added to a petri dish, stirred, and then allowed to stand to form a fibrin plate.

[0055] An equal volume of purified water was added to each oral composition, mixed, and centrifuged to recover the aqueous layer. Next, 50 μl of the resulting aqueous layer was dropped onto the center of the fibrin plate and allowed to stand at 25°C for 3 hours. After leaving the plate for 3 hours, the fibrin plate was photographed with a digital camera, and the area of ​​the dissolved fibrin (dissolved area, mm ) was calculated using image analysis software (WinROOF2018, manufactured by Mitani Shoji Co., Ltd.). 2 ) was calculated. In addition, the ratio to Comparative Example 1 was also calculated according to the following formula: Calculated.

number

[0056] The results are shown in Table 1. When oral compositions containing nattokinase and an oil agent did not contain any amino acids, the activity of nattokinase after storage was low (Comparative Example 1). In contrast, oral compositions containing nattokinase and an oil agent together with isoleucine, glutamine, methionine, tyrosine, tryptophan, phenylalanine, arginine, lysine, histidine, sodium aspartate, sodium glutamate, glycine, or proline improved the initial activity value of nattokinase and the activity value after storage (Examples 1 to 13). Furthermore, oral compositions containing isoleucine, glutamine, methionine, tyrosine, tryptophan, phenylalanine, arginine, lysine, histidine, sodium aspartate, or sodium glutamate as an amino acid improved both the initial activity value of nattokinase and the activity value after storage (Examples 1 to 11). In particular, when arginine, aspartic acid, phenylalanine, or lysine was contained, the activity of nattokinase after storage was high (Examples 6 to 8 and 10), and especially when arginine was contained, the difference between the initial activity of nattokinase and the activity after storage was small (Example 7).

[0057] [Table 1]

Claims

[Claim 1] An oral composition comprising (A) nattokinase, (B) an amino acid, and (C) an oil solution.

Images