Composition containing β-caryophyllene

Abstract

The present invention aims to provide a BCP-containing composition that exhibits superior effects, such as improving the absorption rate and absorption efficiency of BCP by addressing its water-soluble characteristics, enhancing storage stability by addressing its susceptibility to oxidation, and reducing its distinctive odor. [Solution] (A) β-caryophyllene; (B) Krill oil; (C) Surfactants; and (D) Glycerin A composition containing the following:

Description

Technical Field

[0001] The present invention relates to a composition containing β-caryophyllene (hereinafter also referred to as BCP).

Background Art

[0002] BCP is a natural component contained in essential oils of plants such as spices and herbs, and is widely used as a fragrance that can also be used as a food. It is also known that BCP is a compound useful for maintaining or improving liver function (Patent Document 1). However, BCP has characteristics such as low water solubility, easy oxidation, and a unique odor, and it has been desired to address these characteristics when ingesting BCP.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] An object of the present invention is to provide a BCP-containing composition that addresses the above-described characteristics that are desired to be improved when ingesting BCP. Specifically, it is an object to provide a BCP-containing composition that exhibits more excellent effects, such as improving the absorption rate and absorption efficiency of BCP by addressing the characteristics related to water solubility, enhancing the storage stability by addressing the characteristic of being easily oxidized, and reducing the unique odor.

Means for Solving the Problems

[0005] As a result of diligent research, the inventors discovered that mixing BCP with krill oil, a surfactant, and glycerin forms an oil-in-water emulsion when mixed with an aqueous medium. Furthermore, they found that the self-emulsifying composition obtained by mixing BCP with krill oil, a surfactant, and glycerin exhibits high absorption rate and absorption efficiency of BCP, high storage stability, and reduced distinctive odor, thus completing the present invention.

[0006] In other words, the present invention is as follows. [1](A)β-caryophyllene; (B) Krill oil; (C) Surfactants; and (D) Glycerin A composition containing the following: [2] The composition according to [1], wherein the weight ratio (B:C:D) of (B) krill oil, (C) surfactant, and (D) glycerin is represented in the following ratio. B:C:D = 4~8:1~4:1~3 [3] The composition according to [1], wherein the weight ratio (B:C:D) of (B) krill oil, (C) surfactant, and (D) glycerin is represented in the following ratio. B:C:D = 4~5:3~4:2~3 [4] The composition according to any one of [1] to [3], wherein the krill oil contains 30% by mass or more of phospholipids and 22% by mass or more of ω3 fatty acids. [5] The composition according to any one of [1] to [3], wherein the krill oil contains 56% by mass or more of phospholipids and 27% by mass or more of ω3 fatty acids. [6] The composition according to any one of [1] to [5], wherein the surfactant is polysorbate 80. [7] The composition according to any one of [1] to [6], wherein (A) β-caryophyllene is contained in a concentration of 2.5% to 20% by mass. An oil-in-water emulsion composition comprising the composition described in any of [8][1] to [7] and an aqueous medium. [9] The oil-in-water emulsion composition according to [8], wherein the total amount of (B) krill oil, (C) surfactant, and (D) glycerin is contained in the oil-in-water emulsion composition at a concentration of 0.002% by mass to 33.3% by mass.

[10] The oil-in-water emulsion composition according to [8] or [9], wherein the weight-average particle size of the emulsion is 50 to 1000 nm. Food and beverages comprising the composition described in any of

[11] , [1], to [7] or the oil-in-water emulsion composition described in any of [8], to

[10] . A pharmaceutical product comprising the composition described in any of

[12] [1] to [7] or the oil-in-water emulsion composition described in any of [8] to

[10] . A cosmetic comprising the composition described in any of

[13] [1] to [7] or the oil-in-water emulsion composition described in any of [8] to

[10] . [Effects of the Invention]

[0007] The present invention addresses the characteristics of BCP, such as its low water solubility, susceptibility to oxidation, and distinctive odor, and provides a BCP-containing composition with high absorption rate and efficiency, high storage stability, and reduced distinctive odor. [Brief explanation of the drawing]

[0008] [Figure 1] Figure 1 is a graph showing the results of evaluating the elution and dispersibility of β-caryophyllene over time when using various self-emulsifying formulation samples (SEDDS / BCP) and β-caryophyllene stock solution prepared in Preparation Example 1. [Figure 2] Figure 2 is a graph showing the results of evaluating the elution and dispersibility of β-caryophyllene at 120 minutes after adding various self-emulsifying formulation samples and β-caryophyllene stock solution prepared in Preparation Example 1 to the test solution. [Figure 3]Figure 3 is a graph showing the results of evaluating the elution and dispersion properties of β-caryophyllene over time when using self-emulsifying formulation samples (Krill oil: Tween80: glycerin = 5:3:2) containing various concentrations of β-caryophyllene and the β-caryophyllene stock solution. [Figure 4] Figure 4 is a photograph showing the results of observing the self-emulsifying formulation sample suspension with an electron microscope. [Figure 5] Figure 5 is a graph showing the particle size distribution when the self-emulsifying formulation sample is dispersed in purified water. [Figure 6] Figure 6 is a graph showing the results of a stability test when the β-caryophyllene stock solution, the self-emulsifying formulation sample, and a self-emulsifying formulation control sample using Medium chain triglyceride (MCT) instead of Krill oil are each placed in an airtight container and stored at a temperature of 40°C for four weeks. [Figure 7] Figure 7 is a graph showing the results of evaluating the absorbability of β-caryophyllene when the β-caryophyllene stock solution and the self-emulsifying formulation sample are each orally administered to rats, using the amount of BCP in plasma as an index. [Figure 8] Figure 8 is a graph showing the results of evaluating the change in blood concentration of Eicosapentaenoic acid when the self-emulsifying formulation sample and purified water as a control are each orally administered to rats. [Figure 9] Figure 9 is a graph showing the results of evaluating the odor of the self-emulsifying formulation (SEDDS) sample, the Krill oil (KO / BCP) sample, and purified water as a control.

Mode for Carrying Out the Invention

[0009] <Composition Containing β-Caryophyllene> The first embodiment of the present invention is (A) β-caryophyllene; (B) Krill oil; (C) Surfactant; and (D) Glycerin A composition comprising

[0010] The composition of this embodiment can be used as a β-caryophyllene composition having self-emulsifying properties by containing at least the above four components (β-caryophyllene, krill oil, surfactant, and glycerin). The self-emulsifying property means the property of forming an oil-in-water emulsion when mixed with an aqueous medium.

[0011] (β-caryophyllene) β-Caryophyllene is a natural volatile component contained in essential oils obtained from plants such as cloves, which are plants of the Myrtaceae family, and is used as a flavor that can also be used in foods.

[0012] β-Caryophyllene is a compound represented by the following formula (1).

Chemical formula

[0013] β-Caryophyllene contains the following isomers and is not particularly limited, but (-)-β-caryophyllene is preferred.

[0014] (-)-β-caryophyllene: (1R,1β,4E,9α)-4,11,11-trimethyl-8-methylene-bicyclo[7.2.0]undeca-4-ene (+)-β-caryophyllene: (1S,4E,9R)-4,11,11-trimethyl-8-methylene-bicyclo[7.2.0]undeca-4-ene

[0015] β-Caryophyllene may be obtained by extraction and / or purification from plants, may be obtained synthetically, or may be a commercially available product. Commercially available products include, for example, those manufactured by Tokyo Chemical Industry Co., Ltd. and FUJIFILM Wako Pure Chemical Corporation.

[0016] β-caryophyllene may be in its free form, or a pharmaceutically acceptable salt or ester. It may also be in the form of the tel element.

[0017] Examples of pharmaceutically acceptable salts of β-caryophyllene include salts with inorganic or organic bases, or salts with basic amino acids. Examples of inorganic bases include alkali metal salts such as sodium and potassium; alkaline earth metal salts such as calcium and magnesium; and ammonium salts and aluminum salts. Examples of organic bases include primary amines such as ethanolamine; secondary amines such as diethylamine, diethanolamine, dicyclohexylamine, and N,N'-dibenzylethylenediamine; and tertiary amines such as trimethylamine, triethylamine, triethanolamine, pyridine, and picoline. Examples of basic amino acids include lysine, arginine, and ornithine.

[0018] Examples of β-caryophyllene esters include β-caryophyllene esters formed by the esterification of the methyl group in β-caryophyllene with a carboxylic acid. Specifically, examples include 14-acetoxy-β-caryophyllene, which is formed by the esterification of the methyl group with a carboxylic acid.

[0019] The plants containing β-caryophyllene mentioned above are not particularly limited, but examples include cloves (Syzygium aromaticum), hemp (Cannabis sativa), rosemary (Salvia rosmarinus), and hops (Humulus lupulus).

[0020] The extraction site is not particularly limited as long as it achieves the effects of the present invention, and may be the whole plant or any necessary part (e.g., flowers, flower heads, flower buds, buds, inflorescences, leaves, branches, branch leaves, rhizomes, root bark, roots, bark, fruits, fruit peels, legumes, seeds, etc.). The extraction site may be, for example, at least one selected from the group consisting of roots, stems, leaves, and flowers, preferably at least one selected from the group consisting of stems, leaves, and flowers, and more preferably at least one selected from the group consisting of stems and flowers.

[0021] The extraction method is not particularly limited as long as it achieves the effects of the present invention, and known methods such as continuous extraction, immersion extraction, and countercurrent extraction can be used, and ordinary extraction methods, purification methods, concentration methods, synthesis methods, drying and powdering methods, etc., can be employed. The extract may be the crude extract as is, or it may be a purified or concentrated extract.

[0022] For example, clove extract can be obtained by immersing the stems and flowers of cloves in water and / or an organic solvent and filtering off the residue; the extract from which the solvent has been removed; or these as fine powders; or by dissolving, dispersing, and diluting the above extract or solvent-removed product with a suitable solvent; commercially available products can also be used.

[0023] In this specification, when using plant extracts, more specifically, the extraction solvent may be water (including hot water), methanol, ethanol, isopropanol, alcohols such as ethylene glycol, 1,3-butylene glycol, and glycerin, esters such as ethyl acetate, ketones such as acetone and methyl ethyl ketone, nitriles such as acetonitrile, ethers such as diethyl ether and tetrahydrofuran, saturated hydrocarbons such as pentane, hexane, cyclopentane, and cyclohexane, aromatic hydrocarbons such as toluene, halogenated hydrocarbons such as dichloromethane and chloroform, and other organic solvents such as dimethylformamide and dimethyl sulfoxide (all of which may contain water), and may be one or any mixture of two of these solvents. Of these solvents, water, ethanol, 1,3-butylene glycol, or a mixture thereof is preferred. The extracts described herein can be obtained from various raw material companies, and they are usually sold with excipients, but are not limited to these.

[0024] For example, the extraction solvent for the clove extract is not limited as long as it achieves the effects of the present invention, but it is preferably water, ethanol, or aqueous ethanol, and more preferably aqueous ethanol.

[0025] The β-caryophyllene content is not particularly limited, but may be, for example, 0.01% by mass or more, 0.05% by mass or more, 0.1% by mass or more, 0.3% by mass or more, 0.5% by mass or more, 1% by mass or more, 2.5% by mass or more, 3% by mass or more, 5% by mass or more, 10% by mass or more, or 20% by mass or more, relative to the total amount of the composition, and may also be 95% by mass or less, 70% by mass or less, 50% by mass or less, 20% by mass or less, 10% by mass or less, 5% by mass or less, 3% by mass or less, or 1% by mass or less. When expressed as a concentration range, the above lower and upper limits can be arbitrarily combined. Preferably, it is 0.01 to 95% by mass, more preferably 0.05 to 70% by mass, even more preferably 0.1 to 50% by mass, and particularly preferably 2.5% to 20% by mass.

[0026] (Krill oil) Krill oil is an oil extracted from Antarctic krill. Typical components of krill oil are phospholipids and omega-3 fatty acids. It may also contain other components such as astaxanthin.

[0027] The concentration of phospholipids in krill oil is not particularly limited, but may be, for example, 30% by mass or more, 40% by mass or more, or 56% by mass or more, and there is no particular upper limit. The omega-3 fatty acids in krill oil are not particularly limited, but may be, for example, 22% by mass or more, or 27% by mass or more, and there is no particular upper limit. Furthermore, the concentrations of phospholipids and omega-3 fatty acids in krill oil may be combined to be phospholipids: 30% by mass or more and omega-3 fatty acids: 22% by mass or more, phospholipids: 40% by mass or more and omega-3 fatty acids: 22% by mass or more, or phospholipids: 56% by mass or more and omega-3 fatty acids: 27% by mass or more.

[0028] The types of omega-3 fatty acids contained in krill oil are not particularly limited, but examples include EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid).

[0029] By using krill oil, the storage stability of the composition of this embodiment can be improved.

[0030] (Surfactants) Surfactants can be divided into numerous groups, including anionic surfactants, cationic surfactants, amphoteric surfactants, and nonionic surfactants, and any group of surfactants may be used in the present invention. Examples of nonionic surfactants include fatty alcohols; glyceryl esters such as natural mono, di, and triglycerides; fatty acid esters of fats and other alcohols (e.g., propylene glycol, polyethylene glycol, sorbitan, and cholesterol); polyoxyethylene sorbitan fatty acid esters; or polyoxyethylene ethers. Among these, polyoxyl stearate 40, polyoxyethylene

[0105] polyoxypropylene[5] glycol, polysorbate, or sodium lauryl sulfate are preferred. As for polysorbates, polysorbate 40 with palmitic acid as the fatty acid, polysorbate 60 with stearic acid as the fatty acid, and polysorbate 80 (Tween 80) with oleic acid as the fatty acid are preferred, with polysorbate 80 being more preferred.

[0031] (Glycerin) Glycerin is a polyhydric alcohol widely used as a food additive, humectant, and lubricant.

[0032] The weight ratio (B:C:D) of (B) krill oil, (C) surfactant, and (D) glycerin in the composition is not particularly limited as long as it does not hinder the self-emulsifying ability of the composition, and for example, B:C:D can be 4-8:1-4:1-3. From the viewpoint of elution into aqueous media, B:C:D is preferably 4-5:3-4:2-3, and more preferably 5:3:2.

[0033] <Oil-in-Water Emulsion Composition> Another embodiment of the present invention is an oil-in-water emulsion composition comprising the above composition and an aqueous medium.

[0034] The aqueous medium is not particularly limited as long as it can form the oil-in-water emulsion composition according to this embodiment, but examples include water, lower alcohols (e.g., ethanol and propanol), and water is preferred. These aqueous media may also be used individually or in mixtures of two or more.

[0035] The oil-in-water emulsion composition may optionally contain additional components, as long as they exert the effects of BCP and / or krill oil.

[0036] In the oil-in-water emulsion composition, the total concentration of (B) krill oil, (C) surfactant, and (D) glycerin is not particularly limited, but may be, for example, 0.002% by mass or more, 0.01% by mass or more, 0.05% by mass or more, 0.1% by mass or more, 0.3% by mass or more, 0.5% by mass or more, 1% by mass or more, 2.5% by mass or more, 3% by mass or more, 5% by mass or more, 10% by mass or more, 20% by mass or more, and may be 95% by mass or less, 70% by mass or less, 50% by mass or less, 33.3% by mass or less, 20% by mass or less, 10% by mass or less, 5% by mass or less, 3% by mass or less, or 1% by mass or less. When expressed as a concentration range, the above lower and upper limits can be arbitrarily combined. For example, it may be 0.002 to 95% by mass, and preferably 0.002% by mass or more to 33.3% by mass.

[0037] The oil-in-water emulsion composition according to this embodiment is easily emulsified, so no treatment is required to form an emulsion. However, if necessary, an emulsifying treatment may be performed, or emulsification may be achieved by mixing at the time of use.

[0038] In oil-in-water emulsion compositions, the weight-average particle size of the emulsion is not particularly limited, but is, for example, 50 to 1000 nm. This particle size is preferable from the viewpoint of bioavailability. The weight-average particle size can be determined, for example, by measuring the particle size distribution and calculating the median diameter using a Zetasizer Nano ZS (Spectris Co., Ltd.).

[0039] The oil-in-water emulsion composition according to this embodiment can improve the absorption rate and absorption efficiency of BCP, thereby increasing the BCP concentration in the blood. Furthermore, its storage stability is improved, allowing it to remain as BCP without decomposition during storage. In addition, the distinctive odor of BCP can be reduced.

[0040] <Food and beverages> Another embodiment of the present invention is a composition containing the above β-caryophyllene or the above oil-in-water emulsion. This is a food and beverage containing a luciol composition.

[0041] The amount of the composition containing β-caryophyllene or the oil-in-water emulsion composition added to food and beverages is not particularly limited as long as it allows the effects of β-caryophyllene and / or krill oil to be exerted, and examples include ordinary foods and beverages (e.g., nutritional supplements, health supplements, nutritional preparations, etc.), health functional foods (i.e., nutrient functional foods, foods for specified health uses, and foods with functional claims), supplements, food additives, etc., and is not particularly limited.

[0042] The types and forms of these foods and beverages are not particularly limited, as long as they can exert the effects of β-caryophyllene and / or krill oil. For example, they may be in liquid, paste, or solid form, and examples include soft drinks, alcoholic beverages, stick pouches, and gummies.

[0043] Supplements may be solid supplements such as capsules, soft capsules, powders, granules, or tablets, or liquid supplements such as solutions, syrups, suspensions, or emulsions.

[0044] Furthermore, the food and beverage according to this embodiment may contain ingredients that are normally added during the manufacture of food and beverages, such as proteins, carbohydrates, fats, nutrients, seasonings, flavorings, and additives. For example, carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose, sucrose, and oligosaccharides; and polysaccharides such as dextrin and cyclodextrin, as well as sugar alcohols such as xylitol, sorbitol, and erythritol.

[0045] The food and beverage according to this embodiment is not particularly limited in terms of the target of intake, intake amount, timing of intake, frequency of intake, etc., as long as it can exert the effects of β-caryophyllene and / or krill oil. For example, the target of intake is typically a healthy person, but may also be a person who does not have a specific disease related to the effects of β-caryophyllene. The intake amount may be appropriately set within the range of 0.001 mg to 10000 mg of β-caryophyllene per day for a human (adult) weighing approximately 60 kg. The timing of intake may be before meals, between meals, after meals, before bedtime, etc. The above intake amount of food and beverage may be taken once a day, or divided into multiple doses per day.

[0046] <Pharmaceuticals> Another embodiment of the present invention is a pharmaceutical product comprising the above-mentioned β-caryophyllene composition or the above-mentioned oil-in-water emulsion composition.

[0047] There are no particular restrictions on the dosage form of the medicine, but examples include capsules, soft capsules, powders, granules, tablets, solutions, syrups, suspensions, emulsions, inhalants, topical preparations, ointments, creams, etc. There are no particular restrictions on the route of administration, but it may be administered orally or parenterally.

[0048] Pharmaceuticals may contain pharmacologically acceptable carriers, and examples of pharmacologically acceptable carriers include commonly used excipients, binders, disintegrants, lubricants, as well as various carriers, stabilizers, surfactants, plasticizers, solubilizers, reducing agents, buffers, sweeteners, bases, adsorbents, flavoring agents, suspending agents, antioxidants, glossing agents, coating agents, humectants, wetting agents, wetting modifiers, fillers, defoamers, cooling agents, colorants, flavoring agents, fragrances, sugar coating agents, isotonic agents, softeners, emulsifiers, viscosity modifiers, foaming agents, pH adjusters, diluents, dispersants, disintegration aids, disintegration extenders, fragrances, moisture-proofing agents, preservatives, preservatives, solvents, solubilizers, solvents, fluidizers, antistatic agents, bulking agents, humectants, and moisturizing agents.

[0049] The pharmaceutical product of this embodiment may contain other active ingredients as desired.

[0050] The content of the β-caryophyllene-containing composition or the oil-in-water emulsion composition in a pharmaceutical product is not particularly limited as long as it is an amount effective in exhibiting the pharmaceutical effect, and generally varies depending on the dosage form, but can be set within a range of, for example, about 0.01% by mass to about 99.9% by mass, so as to achieve the desired dose. The amount of the β-caryophyllene-containing composition or the oil-in-water emulsion composition relative to the total amount of the pharmaceutical product may be 1 ng / mL to 1 mg / mL, or for example, 1 ng / mg to 1 mg / mg.

[0051] The target population, dosage, timing of administration, frequency of administration, and duration of administration are not particularly limited as long as the active ingredient, β-caryophyllene, exerts its effects to be effective in treating and / or preventing specific diseases, and can be appropriately adjusted according to the age, sex, weight, symptoms, therapeutic effect, area of ​​treatment site, dosage form, and method of administration of the target population. It is preferable that the dosage does not interfere with the exertion of the effects of krill oil. For example, the target population may be those with diseases that can be treated and / or prevented by the effects of β-caryophyllene. The dosage may be appropriately set within the range of 0.001 mg to 10000 mg per day as β-caryophyllene for a human (adult) weighing approximately 60 kg. The timing of administration may be before meals, between meals, after meals, before bedtime, etc. The above dosage of medicine may be administered once a day or divided into multiple doses per day. The duration of administration may be, for example, one day or more and one year or less, or may be continued for a longer period.

[0052] <Cosmetics> Another embodiment of the present invention is a cosmetic product comprising the above-mentioned β-caryophyllene composition or the above-mentioned oil-in-water emulsion composition.

[0053] The dosage form of the cosmetic according to this embodiment is not particularly limited, but examples include solutions, suspensions, emulsions, topical preparations, ointments, creams, etc. For example, the effects of β-caryophyllene can be exerted by applying it to the skin, hair, oral cavity, etc.

[0054] The cosmetic according to this embodiment may contain any ingredients that are normally added during cosmetic manufacturing. Examples of such ingredients include oils, emulsifiers, emollients, lipids, humectants, moisturizers, binders, conditioning agents, emulsion stabilizers, preservatives, chelating agents, metal ion sequestering agents, abrasives, pH adjusters, surfactants, fragrances, colorants, and the like.

[0055] The cosmetic of this embodiment may contain other active ingredients as desired.

[0056] The content of a composition containing β-caryophyllene or the oil-in-water emulsion composition in a cosmetic product is not particularly limited as long as the cosmetic effects of β-caryophyllene are exerted. Generally, it varies depending on the dosage form, but can be set within a range of, for example, about 0.01% by mass to about 99.9% by mass, so as to achieve the desired application amount. The amount of the composition containing β-caryophyllene or the oil-in-water emulsion composition relative to the total amount of the cosmetic product may be 1 ng / mL to 1 mg / mL, or for example, 1 ng / mg to 1 mg / mg.

[0057] The target population, dosage, timing, frequency, and duration of application are not particularly limited as long as the active ingredient, β-caryophyllene, exerts its effects to be effective in treating and / or preventing specific diseases, and can be appropriately adjusted according to the target population's age, sex, weight, symptoms, cosmetic effects, application area, dosage form, and application method. It is preferable that the effects do not interfere with the exertion of krill oil. For example, the target population is typically healthy individuals, but may also be those who do not have specific diseases related to the effects of β-caryophyllene. The dosage for a human (adult) weighing approximately 60 kg is β-caryophyllene The dosage may be set appropriately within a range of 0.001 mg to 10,000 mg per day. The timing of application may be in the morning, noon, evening, or before bedtime. The above-mentioned amount of cosmetic may be administered once a day or divided into multiple applications per day. The application period may be, for example, one day or more and one year or less, or it may be applied continuously for a longer period. [Examples]

[0058] The present invention will be described more specifically below based on examples, but the present invention is not limited to the following examples.

[0059] <Preparation Example: Preparation of a β-caryophyllene-containing self-emulsifying formulation> A self-emulsifying drug delivery system (SEDDS) sample was obtained by uniformly mixing (A) β-caryophyllene (BCP, C0796 (product code), Tokyo Chemical Industry Co., Ltd.) (5% by mass) with (B) krill oil (KO, superba Boost, phospholipids: 56% by mass or more and ω3 fatty acids: 27% by mass or more) (C) Tween80; and (D) glycerin (GLY) in the following ratios, each heated to 40°C. Sample 1: Krill oil:Tween80:Glycerin = 4:4:2 Sample 2: Krill oil:Tween80:Glycerin = 4:3:3 Sample 3: Krill oil:Tween80:Glycerin = 5:3:2 Sample 4: Krill oil:Tween80:Glycerin = 5:2:3 Sample 5: Krill oil:Tween80:Glycerin = 6:3:1 Sample 6: Krill oil:Tween80:Glycerin = 6:2:2 Sample 7: Krill oil:Tween80:Glycerin = 7:2:1 Sample 8: Krill oil:Tween80:Glycerin = 7:1:2 Sample 9: Krill oil:Tween80:Glycerin = 8:1:1 Sample 10: Krill oil:Tween80:Glycerin = 8:0:2 Sample 11: Krill oil:Tween80:Glycerin = 10:0:0

[0060] <Example 1: Evaluation of the elution and dispersibility of β-caryophyllene> For the various self-emulsifying formulation samples and β-caryophyllene stock solution prepared in Preparation Example 1, 5 mg of β-caryophyllene was added to the test solution, and an elution test was performed using the paddle method under the following conditions. For sample collection as described below, samples were collected using a syringe fitted with a needle. Subsequently, an equal volume of acetonitrile was mixed in to suppress the precipitation of β-caryophyllene during the test, and the quantitative sample was prepared. β-caryophyllene in the quantitative sample was quantified using high-performance liquid chromatography (HPLC)-UV under the following analytical conditions.

[0061] (Dissolution test method) Equipment: NTR-6100A (Toyama Sangyo Co., Ltd.) Test solution: Purified water Test solution volume: 100 mL Stirring speed: 50rpm Temperature: 37℃ Sample collection: 5, 10, 15, 30, 60, 120 minutes (200 μL)

[0062] (HPLC-UV analysis conditions) Column used: Kinetex® 2.6μm, EVO C18 100A 2.1×50mm (GL Sciences Co., Ltd.) Detector: SPD-M10Avp UV-PDA Detector (Shimadzu Corporation) Pump: LC-10ADvp (Shimadzu Corporation) Mobile phase flow rate: 0.25mL / min Mobile phase: A: 5 mM ammonium acetate aqueous solution B: Acetonitrile A:B = 22:78 Column temperature: 40℃ Retention time: 2.3 minutes

[0063] The results showed that when using the stock solution of β-caryophyllene, or when using the sample containing only β-caryophyllene and krill oil (Sample 11 above), or the sample containing only β-caryophyllene, krill oil, and glycerin (Sample 10 above), β-caryophyllene hardly dissolved into the water even after time had passed (Figures 1 and 2). On the other hand, in all of the samples containing β-caryophyllene, krill oil, Tween80, and glycerin (Samples 1-9 above), β-caryophyllene dissolved and dispersed into the water over time. Among these, Samples 2-3 showed particularly excellent dissolution and dispersibility in water. Therefore, in the following experiment, the weight ratio of krill oil:Tween80:glycerin = 5:3:2, which is the same as in Sample 3 above, was used.

[0064] Furthermore, to investigate the effect of different β-caryophyllene concentrations on elution into water, self-emulsifying formulation samples were obtained by uniformly mixing (A) β-caryophyllene (2.5% by mass, 5% by mass, 10% by mass, or 20% by mass) with (B) krill oil; (C) Tween 80; and (D) glycerin, each heated to 40°C. When elution tests were performed using these samples with the method described above, it was shown that β-caryophyllene eluted into water regardless of the β-caryophyllene concentration (Figure 3).

[0065] <Example 2: Observation of emulsion droplet morphology> A self-emulsifying formulation sample suspension (20 mg / mL) was prepared using purified water as the solvent. 5 μL of this suspension was added to a Formvar membrane-attached mesh (Nisshin EM Co., Ltd.) that had been hydrophilized using an electron microscope hydrophilization treatment device (JEOL Ltd.). After standing for 10 minutes, the mesh was stained with a 25% EM stainer solution and observed using a transmission electron microscope. A Hitachi HT-7600 transmission electron microscope was used and operated at a voltage of 80 kV. As a result, emulsion formation was confirmed (Figure 4).

[0066] <Example 3: Particle size distribution measurement> The aforementioned self-emulsifying formulation sample was dispersed in purified water and diluted with purified water to a concentration of 2 μg / mL to prepare the measurement sample. The particle size distribution was measured using Zetasizer Nano ZS (Spectris Co., Ltd.), and the median diameter was calculated. The results showed a weight-average particle size of 186 nm and a polydispersity index (PDI) of 0.53 (Figure 5).

[0067] <Example 4: Stability Test> The stock solution of β-caryophyllene, a self-emulsifying formulation sample (SEDDS / BCP), and a self-emulsifying formulation control sample (MCT-SEDDS / BCP) using medium chain triglyceride (MCT) instead of krill oil were each placed in airtight containers and stored at 40°C for 4 weeks. The MCT-SEDDS / BCP was used in the same weight ratio as SEDDS / BCP (i.e., MCT:Tween80:glycerin = 5:3:2, BCP:5% by mass). After storage, each sample was dissolved in acetonitrile to prepare samples for quantification. The amount of β-caryophyllene contained in these samples was quantified using HPLC-UV under the same analytical conditions as in Example 1. As a result, the stock solution of β-caryophyllene and the self-emulsifying formulation control sample using MCT showed residual BCP A decrease in BCP was observed (Figure 6). On the other hand, the self-emulsifying formulation sample retained almost 100% of BCP even after 4 weeks. In other words, the self-emulsifying formulation sample was confirmed to exhibit excellent storage stability. It is thought that the antioxidant effect of astaxanthin contained in krill oil had a great effect on storage stability.

[0068] <Example 5: Evaluation of oral absorption of β-caryophyllene in rats> Male SD rats (SLC Japan Co., Ltd.) aged 6-8 weeks were administered orally via gastric tube to 6-8 week old rats (SLC Japan Co., Ltd.) using a gastric tube. Both the stock solution and a self-emulsifying formulation sample were suspended in purified water. At predetermined intervals (15, 30, 45, 1, 2, 4, 6, 12, and 24 hours), approximately 300 μL of blood was collected from the rat tail vein into a heparinized tube and centrifuged at 10,000 × g for 10 minutes. β-caryophyllene was extracted from the plasma obtained after centrifugation using the sample processing method described below to obtain a quantitative sample. β-caryophyllene in the quantitative sample was quantified using gas chromatography-mass spectrometry (GC-MS). Furthermore, to calculate bioavailability, β-caryophyllene was dissolved in DMSO and administered intravenously to rats at a dose of 10 mg / kg (body weight).

[0069] (Sample processing method) 100 μL of plasma sample was placed in a tube, and 75 μL of ethyl acetate solution containing 1,8-cineole (2 μg / mL) as an internal standard was added. The mixture was then shaken for 10 minutes. After shaking, the mixture was centrifuged at 10,000 × g for 10 minutes, and the resulting supernatant was used as the quantitative sample.

[0070] (GC-MS analysis conditions) Column used: HP-5 (length 30m x inner diameter 0.32mm, film thickness 0.25μm) (Agilent Technologies, Inc.) Detector: JMS-Q1000GC (JEOL Ltd.) Mobile phase flow rate: 1.5mL / min Mobile phase: Helium Oven temperature: 60℃ for 0-2 minutes 2-11.5 minutes: 60-250℃ 11.5-13.5 minutes: 250℃ Inlet temperature: 230℃ Injection volume: 1μL Retention time: β-caryophyllene: approximately 6 minutes (molecular weight: 204, quantitative ion: 79) 1,8-Cineole: Approximately 3 minutes (Molecular weight: 154, Quantitative ion: 93)

[0071] As a result, the maximum value of plasma β-caryophyllene (C) max The concentration was 0.3 ± 0.1 μg / mL when using the β-caryophyllene stock solution, compared to 1.5 ± 0.2 μg / mL when using the self-emulsifying formulation sample (Figure 7). Furthermore, the bioavailability (BA) was 1.5% when using the β-caryophyllene stock solution, compared to 7.7% when using the self-emulsifying formulation sample. In other words, it was shown that by using the self-emulsifying formulation sample, β-caryophyllene can be efficiently absorbed into the body when administered orally, and its concentration in plasma can be increased.

[0072] <Example 6: Evaluation of blood concentration profile of Eicosapentaenoic acid> As experimental animals, 6-8 week old SD male rats (Nippon SLC Co., Ltd.) were orally administered a self-emulsifying formulation sample suspended in purified water at a dose of 100 mg β-caryophyllene / kg (body weight) via a gastric tube. Since endogenous EPA is present in the blood, the difference between the rats and the control group was calculated. The control group was orally administered purified water instead of the self-emulsifying formulation sample. At predetermined times (15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, and 24 hours), approximately 300 μL of blood was collected from the rat tail vein into a heparinized tube and centrifuged at 10,000 × g for 10 minutes. The plasma obtained from the centrifuged samples was deproteinized according to the sample processing method described below to obtain quantitative samples. Eicosapentaenoic acid (EPA) in the quantitative samples was quantified using HPLC-UV under the following analytical conditions.

[0073] (Sample processing method) A 50 μL plasma sample was placed in a tube, and 150 μL of acetonitrile containing anthracene (100 ng / mL) as an internal standard was added. The tube was then cooled on ice for 10 minutes. After that, the mixture was centrifuged at 10,000 × g at 4°C for 10 minutes, and the resulting supernatant was filtered through a 0.2 μm filter to obtain the quantitative sample.

[0074] (HPLC-UV analysis conditions) Column used: Inertsil ODS-4 (particle size 3.0 μm, column size 4.6 × 150 mm) (GL Sciences Co., Ltd.) Detector: SPD-10Avp UV-VIS Detector (Shimadzu Corporation) Pump: LC-10ADvp (Shimadzu Corporation) Mobile phase flow rate: 1.0mL / min Mobile phase: A: 0.1% formic acid B: 0.1% acetonitrile A:B = 15:85 Column temperature: 40℃ Retention time: 4.7 minutes

[0075] As a result, the maximum value of plasma EPA (ΔC max The self-emulsifying formulation sample showed an increase of 2.9 ± 0.1 μg / mL compared to the control group (Figure 8). Furthermore, the area under the plasma EPA concentration change-time curve (ΔAUC) in the self-emulsifying formulation sample was also shown. 0-24h The concentration was 16.7 ± 3.4 μg·h / mL. In other words, by using a self-emulsifying formulation sample, it was possible to confirm the increase in plasma concentration due to EPA derived from krill oil administration when administered orally.

[0076] <Example 7: Odor Evaluation> Self-emulsifying formulation (SEDDS) samples and krill oil (KO / BCP) were each added at a ratio of 1 g (equivalent to the amount of krill oil) to 100 mL of purified water and stirred at 50 rpm. No samples were added to the control group. Odor measurements were taken 5 minutes after addition using XP-329III. RThis was done using (Shin-Cosmos Electric Co., Ltd.). As a result, it was confirmed that the odor level was reduced in the self-emulsifying formulation (SEDDS) sample compared to krill oil (KO / BCP) (Figure 9). In other words, it was shown that the amount of BCP present on the water surface decreased due to the dispersion of BCP in the water by SEDDS, and as a result, the amount of BCP diffused into the atmosphere decreased, leading to a reduction in odor.

Claims

1. (A) β-caryophyllene; (B) Krill oil; (C) Surfactants; and (D) Glycerin A composition containing the following:

2. The composition according to claim 1, wherein the weight ratio (B:C:D) of (B) krill oil, (C) surfactant, and (D) glycerin in the composition is expressed in the following ratio. B:C:D=4~8:1~4:1~3

3. The composition according to claim 1, wherein the weight ratio (B:C:D) of (B) krill oil, (C) surfactant, and (D) glycerin in the composition is expressed in the following ratio. B:C:D=4~5:3~4:2~3

4. The composition according to claim 1, wherein the krill oil contains 30% by mass or more of phospholipids and 22% by mass or more of ω3 fatty acids.

5. The composition according to claim 1, wherein the krill oil contains 56% by mass or more of phospholipids and 27% by mass or more of ω3 fatty acids.

6. The composition according to claim 1, wherein the surfactant is polysorbate 80.

7. The composition according to claim 1, wherein (A) β-caryophyllene is contained in the composition at a concentration of 2.5% to 20% by mass.

8. An oil-in-water emulsion composition comprising the composition described in claim 1 and an aqueous medium.

9. The oil-in-water emulsion composition according to claim 8, wherein the total amount of (B) krill oil, (C) surfactant, and (D) glycerin is contained in the oil-in-water emulsion composition at a concentration of 0.002% by mass to 33.3% by mass.

10. The oil-in-water emulsion composition according to claim 8, wherein the weight-average particle size of the emulsion is 50 to 1000 nm.

11. Food and beverages comprising the composition according to any one of claims 1 to 7 or the oil-in-water emulsion composition according to any one of claims 8 to 10.

12. A pharmaceutical product comprising the composition according to any one of claims 1 to 7 or the oil-in-water emulsion composition according to any one of claims 8 to 10.

13. A cosmetic comprising the composition according to any one of claims 1 to 7 or the oil-in-water emulsion composition according to any one of claims 8 to 10.

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