External or internal composition

Abstract

To provide a composition capable of improving a functionality of flavonoid, such as nobiletin.SOLUTION: Provided is a combination of flavonoid, such as nobiletin, and coumarin, such as auraptene, to improve a functionality, such as an anti-inflammatory effect, of the flavonoid.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an external or internal composition. More specifically, the present invention relates to an external or internal composition with improved functionality by flavonoids such as nobilitin.

Background Art

[0002] Fruits contain functional components such as carotenoids, flavonoids, limonoids, coumarins, and polyphenols. Such functional components are incorporated into health foods such as functional foods for the purpose of increasing the added value of foods.

[0003] Among such functional components, nobilitin, which is abundant in mandarins and oranges, is cited as an example of flavonoids. Nobilitin has been reported to have an effect of suppressing reactions related to carcinogenesis, arteriosclerosis, and inflammation, and furthermore, research on the prevention and improvement of anti-dementia has also been conducted (Non-Patent Document 1). In addition, auraptene contained in buntan and karatachi is cited as an example of coumarin. Auraptene has been mainly studied for its cancer-suppressing effect, and in addition, an effect of activating lipid metabolism and an effect of suppressing blood coagulation reaction have been reported (Non-Patent Document 1).

Prior Art Documents

Non-Patent Documents

[0004]

Non-Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] Although nobilitin has been reported to have functionality such as an anti-inflammatory effect, its action alone is not sufficient. In order to obtain sufficient action effects when incorporated into foods, a large amount of nobilitin is required, resulting in poor cost efficiency.

[0006] Therefore, an object of the present invention is to provide a composition capable of improving the functionality of flavonoids such as nobilitin.

Means for Solving the Problems

[0007] As a result of intensive studies by the present inventors, it has been found that the functionality of flavonoids such as nobilitin can be improved by combining them with coumarins such as auraptene. The present invention has been completed based on this finding through further studies.

[0008] That is, the present invention provides an invention in the following aspects. Item 1. The following formula (1):

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Advantages of the Invention

[0009] According to the present invention, a composition capable of improving the functionality of flavonoids such as nobilitin is provided.

Brief Description of the Drawings

[0010]

Figure 1

Modes for Carrying Out the Invention

[0011] The external or internal composition of the present invention is characterized by containing a predetermined flavonoid and coumarin. Hereinafter, the external or internal composition of the present invention will be described in detail.

[0012] Predetermined flavonoid (Compound 1) The predetermined flavonoid contained in the external or internal composition of the present invention is Compound 1 represented by the following formula (1).

[0013]

Chemical formula

[0014] In formula (1), R 1 ~R 7 each independently represents a hydrogen atom, a hydroxyl group, an alkyl group or an alkoxy group. Examples of the alkyl group include C1-6 (alkyl group having 1 to 6 carbon atoms) alkyl groups, and examples of the alkoxy group include C1-6 alkoxy groups. Preferred examples of R 1 ~R 7 include a hydrogen atom, a hydroxyl group, and an alkoxy group. More preferred examples include a hydrogen atom, a hydroxyl group, a C1-6 alkoxy group, preferably a hydrogen atom, a hydroxyl group, a C1-4 alkoxy group, and more preferably a hydrogen atom, a hydroxyl group, a C1-2 alkoxy group.

[0015] The above Compound 1 may be used alone or in combination of two or more.

[0016] Particularly preferred examples of the above Compound 1 include nobiletin in which R 1 ~R 7 are all methoxy groups.

[0017] In the present invention, the compounds of formula (I) also include salts. Such salts are not particularly limited as long as they are physiologically acceptable salts. For example, alkali metal salts such as sodium and potassium; alkaline earth metal salts such as calcium; magnesium salts; ammonium salts; salts with organic bases such as ethylenediamine, methylamine, dimethylamine, trimethylamine, dicyclohexylamine, tris(hydroxymethyl)aminomethane, N,N-bis(hydroxyethyl)piperazine, 2-amino-2-methyl-1-propanol, diethanolamine, ethanolamine, N-methylglucamine, L-glucamine, etc.; salts with basic amino acids such as lysine, δ-hydroxylysine, arginine, etc.; salts of mineral acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; salts with organic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, acetic acid, propionic acid, tartaric acid, fumaric acid, maleic acid, malic acid, oxalic acid, succinic acid, citric acid, benzoic acid, mandelic acid, cinnamic acid, lactic acid, glycolic acid, glucuronic acid, ascorbic acid, nicotinic acid, salicylic acid, etc.; or salts with acidic amino acids such as aspartic acid, glutamic acid, etc.

[0018] Predetermined coumarin (Compound 2) The predetermined coumarin contained in the external or internal composition of the present invention is Compound 2 represented by the following formula (2).

[0019]

Chemical formula

[0020] In formula (2), R 8represents a hydrogen atom, a hydroxyl group, an alkyl group, an alkoxy group, or an ester group-containing group. Examples of the alkyl group include C1-C6, preferably C1-C4, and more preferably C1-C2 alkyl groups. Examples of the alkoxy group include C1-C6, preferably C1-C4, and more preferably C1-C2 alkyl groups. Examples of the ester group-containing group include an alkyloxycarbonylalkyl group, and examples of the alkyloxycarbonylalkyl group include C1-C6 (preferably C1-C4, more preferably C1-C2) alkyloxycarbonyl C1-C6 (preferably C1-C4, more preferably C1-C2) alkyl groups.

[0021] In formula (2), R 9 represents a hydrogen atom, a hydroxyl group, an alkyl group, an alkoxy group, a carboxyl group, an aryl group or a coumarinyl group. Examples of the alkyl group include C1-C6, preferably C1-C4, and more preferably C1-C2 alkyl groups. Examples of the alkoxy group include C1-C6, preferably C1-C4, and more preferably C1-C2 alkyl groups. Examples of the aryl group include a phenyl group, a naphthyl group, etc., and preferably a phenyl group. Examples of the coumarinyl group include a 3'-coumarinyl group, a 4'-coumarinyl group, and preferably a 3'-coumarinyl group.

[0022] In formula (2), R 10 represents a hydrogen atom, a hydroxyl group, an alkyl group, an alkoxy group, a carboxyl group or a carboxyalkyl group. Examples of the alkyl group include C1-C6, preferably C1-C4, and more preferably C1-C2 alkyl groups. Examples of the alkoxy group include C1-C6, preferably C1-C4, and more preferably C1-C2 alkyl groups. Examples of the carboxyalkyl group include carboxy C1-C6 (preferably C1-C4, more preferably C1-C2) alkyl groups.

[0023] In formula (2), R 11represents a hydrogen atom, an alkyl group, or an olefinically unsaturated double bond-containing group. Examples of the alkyl group include C1-C15 alkyl groups. Examples of the olefinically unsaturated double bond-containing group include alkenyl groups [e.g., C2-C30, preferably C2-C16 alkenyl groups], alkadienyl groups [e.g., C4-C30, preferably C4-C16 alkadienyl groups such as 2,4-alkadienyl groups and 2,6-alkadienyl groups], alkatrienyl groups [e.g., C6-C30, preferably C6-C16 alkatrienyl groups], alkatetraenyl groups [e.g., C8-C30, preferably C8-C16 alkatetraenyl groups], alkapentaenyl groups [e.g., C10-C30, preferably C10-C16 alkapentaenyl groups], and the like.

[0024] In the present invention, preferred examples of Compound 2 include the compounds shown in the following (21)-(28).

[0025]

Chemical formula

[0026]

Chemical formula

[0027]

Chemical formula

[0028]

Chemical formula

[0029]

Chemical formula

[0030]

Chemical formula

[0031] [Chemistry]

[0032] [Chemistry]

[0033] The above compound 2 may be used alone or in combination of two or more.

[0034] Particularly preferred examples of the above compound 2 include auraptene represented by formula (21).

[0035] In the internal or external composition of the present invention, the mixing ratio of compound 1 and compound 2 is not particularly limited. For example, the amount of compound 2 mixed with 1 mol of compound 1 may be 0.1 to 10 mol. From the viewpoint of further improving the functionality such as anti-inflammatory properties by compound 1 more efficiently, the amount of compound 2 mixed with 1 mol of compound 1 is preferably 0.15 to 5 mol, more preferably 0.2 to 4 mol, still more preferably 0.25 to 2.5 mol, even more preferably 0.25 to 2 mol, still even more preferably 0.25 to 1.5 mol, and particularly preferably 0.25 to 1 mol.

[0036] Other components The internal or external composition of the present invention may consist only of the above-mentioned Compound 1 and Compound 2 as active ingredients, or may further contain additives and / or bases according to the dosage form and use. Such additives and bases are not particularly limited as long as they are pharmaceutically acceptable. For example, excipients, binders, disintegrants, lubricants, isotonic agents, plasticizers, dispersants, emulsifiers, solubilizers, wetting agents, stabilizers, suspending agents, adhesives, gelling agents, coating agents, brightening agents, water, fats and oils, waxes, hydrocarbons, fatty acids, higher alcohols, esters, water-soluble polymers, surfactants, metal soaps, lower alcohols, polyhydric alcohols, pH adjusters, buffers, antioxidants, ultraviolet protectants, preservatives, flavoring agents, fragrances, powders, thickeners, coloring agents, chelating agents, sweeteners, etc. may be mentioned. These additives may be used alone or in combination of two or more. Also, the content of these additives and bases is appropriately set according to the type of additives and bases used, the dosage form and use of the internal or external composition of the present invention, and the like.

[0037] In addition to the above-mentioned Compound 1 and Compound 2 as active ingredients, the internal or external composition of the present invention may optionally contain other nutritional components and pharmacological components. Such nutritional components and pharmacological components are not particularly limited as long as they are pharmaceutically acceptable. For example, antacids, stomachics, digestive agents, intestinal regulators, antispasmodics, mucosal repair agents, anti-inflammatory agents, astringents, antiemetics, antitussives, expectorants, anti-inflammatory enzyme agents, sedative hypnotics, antihistamines, caffeine-like substances, cardiotonic diuretics, antibacterial agents, vasoconstrictors, vasodilators, local anesthetics, crude drug extracts, vitamins, menthol-like substances, etc. may be mentioned. These nutritional components and pharmacological components may be used alone or in combination of two or more. Also, the content of these components is appropriately set according to the type of components used, the dosage form and use of the internal or external composition of the present invention, and the like.

[0038] Formulation form Regarding the dosage forms of the internal or external composition of the present invention, there is no particular limitation as long as oral administration is possible for the internal composition and transdermal or transmucosal administration is possible for the external composition.

[0039] For example, dosage forms of the internal composition include solid preparations such as powders, fine granules, granules (including dry syrups), tablets, pills, capsules (soft capsules, hard capsules), etc.; semi-solid preparations such as jelly preparations; and liquid preparations such as solutions, suspensions, syrups, etc. Further, dosage forms of the external composition include creams, lotions, gels, emulsions, solutions, patches, aerosols, ointments, packs, etc.

[0040] To prepare the internal or external composition of the present invention in these dosage forms, the above-mentioned Compound 1 and Compound 2, which are active ingredients, and additives, bases, and / or pharmacological components added as necessary may be used to formulate according to ordinary formulation techniques.

[0041] Use The use of the internal or external composition of the present invention is not particularly limited as long as it is for internal or external use.

[0042] In the case of the internal composition, for example, pharmaceuticals and food and drink products can be mentioned, and preferably food and drink products. Specific examples of food and drink products include health foods, functional foods, dietary supplements, foods for specified health use, etc. Further, specific forms of food and drink products include supplements (tablets, hard capsules, soft capsules, etc.), beverages (soft drinks, carbonated drinks, beauty drinks, nutritional drinks, fruit drinks, milk drinks, etc.), and concentrated stock solutions of the beverages and powders for preparation at the time of use, etc.

[0043] In the case of the external composition, preferably, pharmaceuticals, quasi-drugs, and cosmetics can be mentioned. Specific examples of these pharmaceuticals, quasi-drugs, and cosmetics include creams, lotions, gels, emulsions, packs, sprays, ointments, packs, cleansing agents, etc. Further, application sites include the skin, mucous membranes, hair, etc.

[0044] The internal or external composition of the present invention can improve the functionality of Compound 1, and thus can be used for the purpose of obtaining the effects based on the functionality of Compound 1. Preferably, the internal or external composition of the present invention can be used as an anti-inflammatory agent. Also, the internal or external composition of the present invention can also be used as an IL-6 production inhibitor, a TNF-α production inhibitor, and / or a NO production inhibitor.

[0045] Dosage and administration Regarding the dosage of the internal or external composition of the present invention, it is appropriately set according to the type of active ingredient, the age, sex, constitution, etc. of the administration subject. For example, as the dosage (oral dosage or intake amount) of the internal composition, for humans per day (in the case of a body weight of 60 kg), the amount of the above Compound 1 is, for example, about 0.1 to 500 mg. Since the internal or external composition of the present invention can improve the functionality of Compound 1, even a small amount that Compound 1 alone cannot fully exhibit functionality can effectively obtain the action effects based on the functionality. From such a perspective, preferable examples of the dosage of the internal composition are, for humans per day (in the case of a body weight of 60 kg), the amount of the above Compound 1, preferably about 0.1 to 100 mg, more preferably about 0.2 to 50 mg, and still more preferably about 0.5 to 20 mg. The internal or external composition of the present invention may be taken internally or applied externally 1 to 3 times a day, preferably 2 or 3 times.

Examples

[0046] Hereinafter, the present invention will be specifically described by way of examples, but the present invention is not limited to these examples.

[0047] (1) Test method (1-1) Cell culture Using the mouse macrophage cell line RAW264.7 cells, cultured in a 10% FBS-DMEM medium (containing 100 U / mL penicillin and 100 μg / mL streptomycin) in a carbon dioxide incubator under conditions of 37 °C, 5% CO2, and humidification. Before the test, the RAW264.7 cells adhered to the culture dish were detached with a cell scraper and prepared with 10% FBS-DMEM medium using a cell counter to obtain the desired number of cells.

[0048] (1-2) Test method RAW264.7 cells were seeded in a 96-well culture plate at a density of 2×10 4 cells / well. After pre-culture, as the test samples, dimethyl sulfoxide (DMSO) solutions of nobiretin and / or auraptene were added. For nobiretin and / or auraptene, they were added at the final concentrations shown in Tables 1-5, and for dimethyl sulfoxide, it was added at a final concentration of 0.1%. Then, the cells were cultured for 24 hours.

[0049] (1-3) Evaluation method for anti-inflammatory effect After the culture was completed, the production amounts of interleukin (IL)-6 and tumor necrosis factor (TNF-α) in the collected culture supernatant were quantified by the enzyme-linked immunosorbent assay (ELISA) to evaluate the anti-inflammatory effect on macrophages in a state of excessive inflammation. In addition, the production amount of nitric oxide (NO) was quantified by the Griess method. Taking the production amounts of IL-6, TNF-α, and NO in the control (Comparative Example 1) as 100% respectively, the relative amounts (%) in each test sample were derived. The smaller these relative amounts are, the better the effect can be evaluated.

[0050] In addition, based on the following formula, the ratio of the IL-6 production inhibition amount obtained in the examples to the IL-6 production inhibition amount equivalent to the additive effect (hereinafter referred to as "the effect of synergistically improving the IL-6 production inhibitory effect") was calculated.

[0051]

Number

[0052] Similarly, based on the following formula, the ratio of the TNF-α production inhibitory amount obtained in the examples to the TNF-α production inhibitory amount equivalent to the additive effect (hereinafter referred to as "the effect of synergistically improving the TNF-α production inhibitory effect") and the ratio of the NO production inhibitory amount obtained in the examples to the NO production inhibitory amount equivalent to the additive effect (hereinafter referred to as "the effect of synergistically improving the NO production inhibitory effect") were calculated.

[0053]

Number

[0054]

Number

[0055] The results regarding the IL-6 production inhibitory effect are shown in Tables 1 to 3, the results regarding the TNF-α production inhibitory effect are shown in Table 4, and the results regarding the NO production inhibitory effect are shown in Table 5.

[0056] Also, to confirm the toxicity of the test sample to RAW264.7 cells, after collecting the supernatant, the WST-8 solution was added to make it 10%, and it was incubated at 37°C for 40 minutes. Then, the cell viability was measured by measuring the absorbance at 40 nm, and the relative viability (%) was calculated with the average value in Comparative Example 1 as 100%. The results are shown in Figure 1.

[0057]

Table 1

[0058]

Table 2

[0059]

Table 3

[0060]

Table 4

[0061]

Table 5

[0062] Regarding the inhibitory effect on IL-6 production (Tables 1 to 3), nobiretin alone was not sufficient (Comparative Examples 2, 6, and 7), and auraptene did not show any or almost no effect depending on the concentration (Comparative Examples 3 and 4), and even when it showed an effect, it was not sufficient (Comparative Example 5). On the other hand, when nobiretin and auraptene were combined, the inhibitory effect on IL-6 production was improved (Examples 1 to 9).

[0063] In addition, the confirmed inhibitory effect on IL-6 production differed depending on the added concentration of each component (Tables 1, 2, and 3). In particular, when the ratio of auraptene to nobiretin was as shown in Examples 1, 2, 4, to 9, a particularly remarkable and excellent effect exceeding the additive effect of the effects of nobiretin and auraptene alone was obtained.

[0064] Furthermore, when comparing Example 1 and Example 2 with respect to the inhibitory effect on IL-6 production, the amount of IL-6 production inhibition was higher in Example 2, but the effect of synergistically improving the IL-6 production inhibitory action was significantly higher in Example 1. Also, when comparing Examples 4 to 6, the amount of IL-6 production inhibition increased in the order of Examples 4, 5, and 6, but for the effect of synergistically improving the IL-6 production inhibitory action, it was higher in the order of Examples 6, 5, and 4, and Example 4 was significantly higher. Furthermore, when comparing Examples 7 to 9, the amount of IL-6 production inhibition increased in the order of Examples 7, 8, and 9, but for the effect of synergistically improving the IL-6 production inhibitory action, it was higher in the order of Examples 9, 8, and 7 (Examples 7 and 8 were equivalent).

[0065] Regarding the TNF-α production inhibitory effect (Table 4), there is room for improvement with nobiretin alone (Comparative Example 2). Regarding auraptene, depending on the concentration, there was no effect at all (Comparative Examples 3 and 4), and even when an effect was shown, it was not sufficient (Comparative Example 5). On the other hand, when nobiretin and auraptene were combined, the TNF-α production inhibitory effect was improved (Examples 1 to 3).

[0066] Furthermore, when comparing Examples 1 to 3 regarding the TNF-α production inhibitory effect, when nobiretin and auraptene were combined at the ratios of Examples 1 and 2, a particularly remarkable effect exceeding the additive effect of the effects of nobiretin and auraptene alone was obtained. Also, when comparing Example 1 and Example 2, although the amount of TNF-α production inhibition was higher in Example 2, the ratio of the amount of TNF-α production inhibition (the effect of synergistically improving the TNF-α production inhibitory action) was higher in Example 1.

[0067] Regarding the NO production inhibitory effect (Table 5), there was almost no effect with nobiretin alone (Comparative Example 8). Regarding auraptene, depending on the concentration, there was no effect at all (Comparative Example 3), and even when an effect was shown, it was not sufficient (Comparative Example 4). On the other hand, when nobiretin and auraptene were combined, the NO production inhibitory effect was improved (Examples 10 and 11).

[0068] Furthermore, when comparing Examples 10 and 11 regarding the NO production inhibitory effect, not only was the amount of NO production inhibition higher in Example 10, but the effect of synergistically improving the NO production inhibitory action was also significantly higher in Example 10.

[0069] In addition, as shown in Figure 1, it was also confirmed that the test sample had no cytotoxicity.

Claims

1. An external or internal composition containing nobilitin and auraptene as active ingredients, containing 0.2 to 2.5 moles of auraptene per 1 mole of nobilitin, and used as an IL6 production inhibitor and / or an NO production inhibitor.

2. The composition according to claim 1, which is a food or drink.

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