Oral compositions containing nattokinase

Abstract

An object of the present invention is to provide a technology for inhibiting the decline in nattokinase activity over time and providing excellent storage stability in an oral composition containing nattokinase. [Solution] An oral composition containing nattokinase together with black ginger extract, hesperidin, glycosyl hesperidin, α-linolenic acid, and / or rutin can inhibit the decrease in nattokinase activity during storage and has excellent storage stability.

Description

[Technical Field]

[0001] The present invention relates to an oral composition containing nattokinase and having improved storage stability. [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 rediscovered 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 foods containing nattokinase are useful for self-medication because they can be easily taken daily without the need for medical supervision.

[0004] Various oral compositions using nattokinase 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 health benefits derived from the milk fermentation product and nattokinase.

[0005] On the other hand, oral compositions containing nattokinase are required to have improved storage stability in order to suppress a decrease in fibrinolytic activity over time, but in the prior art, there has been no sufficient research into techniques for improving the storage stability of oral compositions containing nattokinase. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] WO 02 / 76240 Summary of the Invention [Problem to be solved by the invention]

[0007] An object of the present invention is to provide a technology for inhibiting the decline in nattokinase activity over time and providing excellent storage stability in an oral composition containing nattokinase. [Means for solving the problem]

[0008] The present inventors have conducted extensive research to solve the above-mentioned problems and have found that an oral composition containing nattokinase together with black ginger extract, hesperidin, glycosyl hesperidin, α-linolenic acid, and / or rutin can inhibit the activity of nattokinase from decreasing during storage and has excellent storage stability. The present invention was completed based on this finding and through further research.

[0009] That is, the present invention provides the following aspects. Item 1. An oral composition comprising (A) nattokinase and (B) at least one member selected from the group consisting of black ginger extract, hesperidin, transglycosylated hesperidin, α-linolenic acid, and rutin. Item 2. The oral composition according to Item 1, which is a food product. Item 3. A method for suppressing a decrease in nattokinase activity, comprising allowing (A) nattokinase to coexist with (B) at least one selected from the group consisting of black ginger extract, hesperidin, transglycosylated hesperidin, α-linolenic acid, and rutin. Item 4. An inhibitor of the decline in nattokinase activity, comprising as an active ingredient at least one selected from the group consisting of black ginger extract, hesperidin, glycosyl hesperidin, α-linolenic acid, and rutin. [Effects of the Invention]

[0010] According to the present invention, an oral composition containing nattokinase can be provided with excellent storage stability by suppressing the decrease in nattokinase activity over time, thereby making it possible to provide an oral composition that can exert an excellent fibrinolytic effect. DETAILED DESCRIPTION OF THE INVENTION

[0011] 1. Oral Composition The oral composition of the present invention is characterized by containing nattokinase (sometimes referred to as component (A)) and at least one member selected from the group consisting of black ginger extract, hesperidin, transglycosylated hesperidin, α-linolenic acid, and rutin (sometimes referred to as component (B)). The oral composition of the present invention is described in detail below.

[0012] [(A) Nattokinase] The oral composition of the present invention contains nattokinase, an enzyme having fibrinolytic activity 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 can be incorporated into an oral composition. For example, when using nattokinase obtained by culturing Bacillus subtilis natto, it may be an extract of the culture of Bacillus subtilis natto. Furthermore, nattokinase may be 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. Alternatively, nattokinase may be 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] The content of nattokinase in the oral composition of the present invention may be appropriately determined depending on the form of the oral composition, and may be, for example, 1 to 60,000 FU / g, preferably 200 to 20,000 FU / g, and more preferably 300 to 10,000 FU / g. Note that, in the present invention, the "FU" indicating the activity of nattokinase is a fibrinolytic activity unit according to the standards for Bacillus subtilis natto culture extract foods published by the Japan Health and Nutrition Food Association on January 15, 2003.

[0017] [(B) Black ginger extract, hesperidin, glycosyl hesperidin, α-linolenic acid, and / or rutin] The oral composition of the present invention contains black ginger extract, hesperidin, glycosyl hesperidin, α-linolenic acid, and / or rutin, which can inhibit the decline in nattokinase activity over time and provide excellent storage stability.

[0018] In the present invention, component (B) may be one selected from black ginger extract, hesperidin, glycosyl hesperidin, α-linolenic acid, and rutin and used alone, or two or more of these may be used in combination.

[0019] Among these components (B), from the viewpoint of more effectively suppressing the decrease in nattokinase activity due to storage, preferred are hesperidin, transglycosylated hesperidin, and α-linolenic acid, and more preferred are transglycosylated hesperidin and α-linolenic acid.

[0020] The composition and content of each component of component (B) will be described below.

[0021] (Black ginger extract) Black ginger extract is a component obtained by performing an extraction process using black ginger as an extraction raw material. Black ginger, also known as black ginger, is a plant (Kaempferia parviflora) of the Zingiberaceae family.

[0022] The part of black ginger used as the raw material for extraction is not particularly limited, but preferably the rhizome. The rhizome of black ginger to be subjected to extraction treatment may be subjected to treatment such as drying, cutting, or crushing as necessary to increase the extraction efficiency.

[0023] The extraction treatment may be any common extraction method used in the production of plant extracts, such as solvent extraction treatment, supercritical extraction treatment, steam distillation treatment, etc. Among these, solvent extraction treatment is preferred.

[0024] Examples of extraction solvents used in solvent extraction include water; lower alcohols such as methanol and ethanol; polyhydric alcohols such as propylene glycol and 1,3-butylene glycol; hydrophilic solvents such as acetone; and mixed solvents thereof. Among these extraction solvents, water, lower alcohols, and mixed solvents thereof are preferred, more preferably water, ethanol, and mixed solvents thereof, and even more preferably a mixed solvent of water and ethanol. In the mixed solvent of water and lower alcohol, the ratio of water to lower alcohol is not particularly limited, but may be, for example, about 5 to 90 wt %, preferably about 20 to 80 wt %, and more preferably about 40 to 80 wt %.

[0025] The solvent extraction treatment may be carried out by immersing the target part of black ginger in an extraction solvent and stirring as necessary, for example, by immersing the target part of black ginger in about 0.3 to 20 L, preferably about 0.5 to 1.5 L, of extraction solvent per 100 g of the target part of black ginger for about 0.5 to 24 hours. The temperature condition for the solvent extraction treatment is not particularly limited, but may be, for example, about 40 to 95°C.

[0026] After the extraction process, solids are removed by solid-liquid separation to obtain a black ginger extract (extract). The extract may be purified as needed by filtration or various adsorption processes, such as chromatography using columns packed with carriers such as polystyrene gel (e.g., polystyrene-divinylbenzene copolymer), ion exchange resin, or activated carbon. The extract may be used directly as a non-concentrated extract, or it may be concentrated to produce a soft extract or dried to produce an extract powder.

[0027] In the oral composition of the present invention, the content of the black ginger extract may be appropriately determined depending on the content of nattokinase, the form of the oral composition, etc., and may be, for example, 0.1 to 75 wt %, preferably 0.1 to 70 wt %, more preferably 0.2 to 40 wt %, and even more preferably 0.2 to 10 wt %, calculated as the dry weight of the black ginger extract.

[0028] In addition, in the oral composition of the present invention, the ratio of nattokinase to black ginger extract may be within the range corresponding to the content of each of the above-mentioned components, and for example, the ratio of black ginger extract is 0.25 to 150 mg, preferably 0.25 to 100 mg, more preferably 0.5 to 5 mg, in dry weight terms, per 100 FU of nattokinase.

[0029] (Hesperidin, transglycosylated hesperidin) Hesperidin is a compound in which hesperetin is the aglycone to which rutinose is bound.

[0030] Glycosyl hesperidin is a compound in which one or more glucose residues are α-1,4-linked to a glucose residue in the rutinose unit of hesperidin. In the glycosyl hesperidin, the number of glucoses added to hesperidin is not particularly limited, but may be, for example, 1 to 20, preferably 1 to 10, more preferably 1 to 5, even more preferably 1 to 3, and particularly preferably 1.

[0031] Glycosyl hesperidin can be obtained by, for example, reacting hesperidin with a glycosyltransferase in the presence of a glucose source. Examples of glucose sources include amylose, dextrin, cyclodextrin, maltooligosaccharide, liquefied starch, and gelatinized starch. Examples of glycosyltransferases include α-glycosidase (EC 3.2.1.20), cyclomaltodextrin glucanotransferase (EC 2.4.1.19), and α-amylase (EC 3.2.1.1), which can be selected appropriately depending on the type of glucose source. Specific production methods are described, for example, in the specification of Japanese Patent No. 3060227. Glycosyl hesperidin is commercially available under the trade names "αG Hesperidin H" and "αG Hesperidin PA-T" (both manufactured by Toyo Sugar Refining Co., Ltd.). In the present invention, commercially available glycosyltransperidin can be used.

[0032] In the oral composition of the present invention, the content of hesperidin and / or transglycosylated hesperidin may be appropriately determined depending on the content of nattokinase, the form of the oral composition, etc., and may be, for example, 0.2 to 75 wt %, preferably 0.2 to 70 wt %, more preferably 0.2 to 50 wt %, and even more preferably 0.2 to 10 wt % in terms of the total amount of hesperidin and / or transglycosylated hesperidin.

[0033] In the oral composition of the present invention, the ratio of nattokinase to hesperidin and / or transglycosylated hesperidin may be within the range corresponding to the content of each of the above-mentioned components, and may be, for example, 0.08 to 150 mg, preferably 0.1 to 100 mg, and more preferably 0.1 to 50 mg in total of hesperidin and / or transglycosylated hesperidin per 100 FU of nattokinase.

[0034] (α-linolenic acid) Alpha-linolenic acid is a type of omega-3 fatty acid, a polyunsaturated fatty acid with three double bonds.

[0035] In the present invention, refined α-linolenic acid may be used, but oils and fats containing α-linolenic acid may also be used. For example, vegetable oils such as perilla oil, linseed oil, perilla oil, chia seed oil, and sacha inchi oil are known to be rich in α-linolenic acid, and these vegetable oils may be used as α-linolenic acid.

[0036] In the oral composition of the present invention, the content of α-linolenic acid may be appropriately determined depending on the content of nattokinase, the form of the oral composition, etc., and may be, for example, 0.05 to 40 wt %, preferably 0.2 to 40 wt %, more preferably 0.2 to 25 wt %, and even more preferably 0.2 to 10 wt %.

[0037] In addition, in the oral composition of the present invention, the ratio of nattokinase to α-linolenic acid may be within the range corresponding to the content of each of the above-mentioned components, and for example, the ratio of α-linolenic acid per 100 FU of nattokinase is 0.0075 to 85 mg, preferably 0.05 to 55 mg, more preferably 0.3 to 50 mg, and even more preferably 0.3 to 28 mg.

[0038] (Rutin) Rutin is a type of quercetin glycoside. In the present invention, purified rutin may be used, or a plant extract containing rutin may also be used.

[0039] In the oral composition of the present invention, the content of rutin may be appropriately determined depending on the content of nattokinase, the form of the oral composition, etc., and may be, for example, 0.1 to 50 wt %, preferably 0.2 to 50 wt %, and more preferably 0.2 to 25 wt %.

[0040] In addition, in the oral composition of the present invention, the ratio of nattokinase to rutin may be within the range corresponding to the content of each of the above-mentioned components, and for example, 0.05 to 100 mg, preferably 0.05 to 75 mg, more preferably 0.05 to 50 mg of rutin per 100 FU of nattokinase.

[0041] [Other ingredients] The oral composition of the present invention may contain other nutritional components and pharmacological components in addition to the aforementioned components (A) and (B). 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, minerals, carbohydrates, vegetable oils and fats, fatty acids, flavorings, seasonings, plant extracts (other than black ginger extract), antioxidants, hypoglycemic agents, anticholesterol agents, and immunostimulants. These components may be used alone or in combination of two or more. The content of these components is appropriately determined depending on the types of additives 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 aforementioned components (A) and (B). Such bases and additives are not particularly limited as long as they are usable in foods and pharmaceuticals, and examples thereof include water, lower alcohols, higher alcohols, water-soluble polymers, surfactants, 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 (A) to (C) 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, nutritional supplements, functional foods, and foods for patients. The form of these foods and beverages is not particularly limited, but specific examples include supplements such as capsules (soft capsules and hard capsules), tablets, granules, powders, and jellies; beverages such as energy drinks, soft drinks, carbonated drinks, and lactic acid drinks; and luxury items such as gummies and candies. Among these foods and beverages, supplements are preferred, capsules, tablets, granules, and powders are more preferred, and capsules are particularly preferred, with soft capsules being particularly preferred.

[0046] When the oral composition of the present invention is formulated into an oral pharmaceutical preparation, the aforementioned components (A) to (C) 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, capsules are more preferred, and soft capsules are particularly preferred.

[0047] 2. Method for inhibiting the decline in nattokinase activity and inhibitor for the decline in nattokinase activity The method for suppressing a decrease in nattokinase activity of the present invention is characterized by causing (A) nattokinase to coexist with (B) at least one selected from the group consisting of black ginger extract, hesperidin, transglycosylated hesperidin, α-linolenic acid, and rutin.

[0048] Furthermore, the nattokinase activity decrease inhibitor of the present invention is characterized by containing as an active ingredient at least one selected from the group consisting of black ginger extract, hesperidin, glycosyl hesperidin, α-linolenic acid, and rutin.

[0049] In these methods and agents, the types and amounts of ingredients used, specific embodiments, etc. are as described in the section "1. Oral Composition" above. [Example]

[0050] The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples.

[0051] In the following examples, comparative examples, and formulation examples, the sources, compositions, etc. of the ingredients used are as follows: Nattokinase powder : "Natto Bacillus Culture Extract NSK-SD40" (manufactured by Japan Bioscience Research Institute Co., Ltd.) Black Ginger Extract : "Black Ginger Extract" (Maruzen Pharmaceutical Co., Ltd.), a dried powder of extract from black ginger rhizomes extracted with an aqueous ethanol solution Glycosylated hesperidin : "αG Hesperidin PA-T" (manufactured by Toyo Sugar Refining Co., Ltd.), enzyme-treated hesperidin in which glucose is transferred to hesperidin Vegetable oil containing α-linolenic acid : "Perilla oil" (Oryza Oil & Fat Chemical Co., Ltd.), containing 55% by weight of α-linolenic acid Rutin : "αG Rutin P" (manufactured by Toyo Sugar Refining Co., Ltd.)

[0052] Test Example 1 Oral compositions were prepared with the compositions shown in Table 1. 20 g of each obtained oral composition was filled into a transparent glass bottle (diameter 30 mm, height 65 mm; Maruemu screw tube No. 6) and stored at 40°C or 50°C under light-shielded conditions for 7 days. After storage, nattokinase activity was measured for each oral composition using gelatin-decomposing ability as an index by the following method.

[0053] A gelatin film (thickness 3 mm, area approximately 57 cm) was formed using an aqueous solution (containing 0.2% by weight of methylparaben) in which 20% by weight of gelatin was dissolved. 2) was prepared. 0.3 ml of each oral composition after storage was added to the gelatin coating, and the mixture was left to stand at 25°C for 12 hours. After that, the state of the gelatin coating in the area where the oral composition was added was observed, and nattokinase activity was evaluated according to the following criteria. <Criteria for determining nattokinase activity> 5: The gelatin coating is completely dissolved to the bottom (dissolved to a depth of 3 mm). 4: The gelatin coating has dissolved almost to the bottom (dissolved to a depth of 2.5 mm or more but less than 3 mm). 3: The gelatin coating has dissolved to about half its depth (dissolved to a depth of 1.5 mm or more but less than 2.5 mm). 2: The gelatin film has dissolved slightly (dissolved to a depth of 0.5 mm to less than 1.5 mm). 1: The gelatin film has dissolved to the extent that it has slightly dented (dissolved to a depth of less than 0.5 mm). 0: The gelatin film is not dissolved.

[0054] The results are shown in Table 1. When nattokinase was used alone, its activity significantly decreased after storage (Comparative Example 1). In contrast, when nattokinase was used in combination with black ginger extract, glycosyl hesperidin, α-linolenic acid-containing vegetable oil, or rutin, the decrease in nattokinase activity due to storage was effectively suppressed, and high storage stability was observed (Examples 1 to 7).

[0055] [Table 1]

[0056] Prescription example Soft capsules (containing 300 mg or 400 mg of inner liquid and 150 mg of outer shell per capsule) were produced using inner liquids with the compositions shown in Tables 2 to 5 and capsule shells with the compositions shown in Table 6. The obtained soft capsules were able to effectively prevent the activity of nattokinase from decreasing during storage.

[0057] [Table 2]

[0058] Table 3

[0059] Table 4

[0060] Table 5

[0061] Table 6

Claims

[Claim 1] An oral composition comprising (A) nattokinase and (B) at least one member selected from the group consisting of black ginger extract, hesperidin, transglycosylated hesperidin, α-linolenic acid, and rutin.

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