Composition for alleviating frequent urination

Abstract

To provide a composition capable of alleviating frequent urination.SOLUTION: The present invention provides a composition containing oenothein B as an active ingredient.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a composition for improving frequent urination, which contains oenothein B as an active ingredient. [Background technology]

[0002] Frequent urination is a symptom of an increase in the number of times a day you urinate. Causes of frequent urination include overactive bladder, residual urine, polyuria, urinary tract infection, tumors, and psychological factors. In particular, overactive bladder is a disease that presents with urinary disorders such as urgency to urinate and urge incontinence, and since the symptoms of overactive bladder interfere with daily life and reduce the quality of life, it has been attracting attention in recent years with the aging society.

[0003] Currently, anticholinergic drugs are used in the drug treatment of overactive bladder. Anticholinergic drugs inhibit the action of acetylcholine, which promotes the contraction of the detrusor muscle of the bladder. However, anticholinergic drugs have many problems, such as side effects such as dry mouth and constipation, and attempts are being made to develop more effective methods for preventing and improving overactive bladder. For example, a preventive or improving agent for overactive bladder containing flavonoids selected from isorhamnetin, catechin, hesperetin, quercetin, luteolin, fisetin, chrysin, genistein, and their glycosides as active ingredients (Patent Document 1), and a preventive or improving agent for frequent urination containing at least one active ingredient selected from the group consisting of glucosamine, pyruvic acid, and salts thereof (Patent Document 2) are known. However, the compositions developed so far have not necessarily been effective enough, so there is a demand for the development of a new composition with excellent frequent urination improving effects. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2014-024809 A [Patent Document 2] JP 2016-153384 A Summary of the Invention [Problem to be solved by the invention]

[0005] An object of the present invention is to provide a composition capable of improving frequent urination.

[0006] As a result of extensive research to solve the above problems, the present inventors have surprisingly found that oenothein B improves overactive bladder and improves frequent urination. The present invention has been completed based on this finding.

[0007] The outline of the present invention is as follows. [1] A composition for improving frequent urination containing oenothein B as an active ingredient. [2] A composition for improving overactive bladder, containing oenothein B as an active ingredient. [3] A composition for improving frequent urination described in [1], characterized in that oenothein B is derived from willow herb. [4] A composition for improving overactive bladder described in [2], characterized in that oenothein B is derived from willow herb. Effect of the Invention

[0008] The composition of the present invention has an effect of improving overactive bladder, and therefore exerts an excellent effect of improving frequent urination. [Brief description of the drawings]

[0009] [Figure 1] FIG. 1 is a diagram showing relative values ​​of the expression level of the UPK2 gene in the examples and comparative examples. [Diagram 2] FIG. 2 is a diagram showing the relative expression levels of the UPK3A gene in the examples and comparative examples. [Diagram 3] FIG. 3 is a diagram showing relative values ​​of ROS production amounts in the examples and comparative examples. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] The present invention will be described in detail below. Note that the present invention is not limited to the following embodiments.

[0011] The composition of the present invention is characterized in that it contains oenothein B as an active ingredient. Oenothein B is a type of hydrolyzable tannin and is a compound represented by the following formula (1). Oenothein B is also called oenothein B or oenothein B.

[0012] [ka]

[0013] The oenothein B used in the present invention is not particularly limited as long as it is commonly used in the field of food and beverages, and examples thereof include synthetic products synthesized by methods commonly known to those skilled in the art, plants containing oenothein B, and processed products of said plants. From the viewpoint of safety in oral ingestion, it is preferable to use plants containing oenothein B and processed products of said plants.

[0014] Examples of plants containing othein B used in the present invention include willow herb, eucalyptus, pomegranate, etc. It is preferable to use willow herb or eucalyptus because they are rich in othein B, and it is particularly preferable to use willow herb from the viewpoint of improving frequent urination.

[0015] Examples of processed products of plants containing oenothein B include crushed products, squeezed juice, and extracts, but extracts are preferred because they are rich in oenothein B. The solvent used for extraction may be either a polar solvent or a non-polar solvent, and water, methanol, ethanol, acetone, ethyl acetate, diethyl ether, n-hexane, etc., or a mixture thereof may be used, but from the viewpoint of efficient extraction of oenothein B, it is preferable to use water, hydrous ethanol, or ethanol. The extraction method is not particularly limited as long as it is a method that is usually acceptable for extracting plant components, and examples include solid-liquid extraction methods such as heated extraction and supercritical fluid extraction. The squeezed juice or extract may be in liquid form, but can also be used as a paste or dry powder. When making it into a paste or dry powder, it may be produced using only the product itself, or it may be processed together with an excipient. In addition, the processed product obtained may be further subjected to chemical treatment, enzyme treatment, and purification treatment.

[0016] The content of oenothein B in the composition of the present invention can be measured, for example, by HPLC. For example, it can be measured by an HPLC analyzer equipped with an ultraviolet absorption detector, and the analytical column is CAPCELLPAK C18 UG120 5 μm (4.6×250 mm) manufactured by Osaka Soda Co., Ltd., and the mobile phase is a 0.05% aqueous phosphoric acid solution (mobile phase A) and an acetonitrile solution (mobile phase B). The column temperature can be 40° C., and the flow rate can be 1.0 mL / min. The gradient conditions can be as follows:

[0017] [Table A]

[0018] In addition to oenothein B, other components may be added to the composition of the present invention as necessary. Examples of other components include excipients, lubricants, dietary fibers such as soluble dietary fibers and insoluble dietary fibers, proteins, various vitamins and minerals, microorganisms such as algae and yeasts, etc. Furthermore, as necessary, sweeteners, acidulants, nutritional supplements, stabilizers, binders, gloss agents, thickeners, colorants, diluents, emulsifiers, food additives, seasonings, etc. that are commonly used in the food industry may be mentioned. The content of these other components may be appropriately selected depending on the form of the composition of the present invention.

[0019] As described in the examples below, the composition of the present invention has an effect of promoting uroplakin gene expression and an effect of inhibiting ROS production, and therefore can improve overactive bladder and improve frequent urination. In the present invention, improvement of frequent urination includes both improvement of daytime frequent urination and improvement of nocturnal frequent urination. In this specification, "nighttime" is synonymous with sleeping, and "daytime" refers to time other than sleeping. Since the composition of the present invention can also improve nocturnal frequent urination, it is expected that by reducing the number of times of frequent urination during sleep, it is possible to reduce midnight awakenings and improve the quality of sleep. In addition, in the present invention, "improvement" is used in a sense that includes prevention and treatment (including relief and cure).

[0020] The composition of the present invention has an effect of improving frequent urination, and is therefore used as a composition for improving frequent urination.

[0021] The composition of the present invention is not particularly limited as long as it can be distinguished from other products as a product in terms of its use for improving frequent urination, and for example, the scope of the present invention includes products that display the function of improving frequent urination on the main body, packaging, instructions, or advertising materials of the product of the present invention. For example, so-called health foods such as pharmaceuticals (including quasi-drugs), functional foods such as specified health foods, nutritional functional foods, and functional food whose efficacy has been approved by a designated organization, and feeds, etc. can be mentioned. In addition, even if it is a general food, those that are manufactured and sold with the suggestion of the use of improving frequent urination are included in the scope of the composition for improving frequent urination of the present invention.

[0022] Specifically, a composition for improving frequent urination can be advertised or promoted using phrases such as "relieves the bother of having to urinate," "alleviates the bother of having to urinate," "for those suffering from frequent urination," "prevents overactive bladder," "improves nocturia," "reduces the discomfort of waking up at night to go to the toilet," "reduces the number of times you go to the toilet at night," and the like, and such advertising or promotion can be considered as implementing the present invention.

[0023] The composition of the present invention is not particularly limited as long as it is in a form suitable for oral use, and examples thereof include tablets, capsules, powders, granules, liquids, granules, rods, plates, blocks, solids, rounds, pastes, creams, caplets, gels, chewables, and sticks. Among these, from the viewpoint of ease of administration, tablets, capsules, powders, granules, and liquids are preferred, and tablets, capsules, and granules are more preferred. Examples of oral compositions used as tablets, capsules, powders, granules, and liquids include supplements, food additives, container-packed beverages filled in PET bottles, cans, bottles, etc., and powdered beverages to be dissolved in water (hot water), milk, fruit juice, etc. and drunk.

[0024] The content of onotein B in the composition of the present invention may be appropriately determined within the range in which the effect is exhibited. From the viewpoint of improving frequent urination, the content is preferably 0.001% by mass or more, more preferably 0.01% by mass or more, and particularly preferably 0.1% by mass or more in the solid content. In addition, from the viewpoint of easy continuous oral intake in daily life, the content of onotein B is preferably 80% by mass or less, more preferably 60% by mass or less, even more preferably 40% by mass or less, and particularly preferably 20% by mass or less.

[0025] The amount of intake of the composition of the present invention is not particularly limited, but from the viewpoint of improving frequent urination, the amount of onotein B is preferably 0.01 mg or more per day, more preferably 0.1 mg or more, even more preferably 1 mg or more, and particularly preferably 10 mg or more. Also, from the viewpoint of easy continuous oral intake in daily life, the amount of onotein B is preferably 1000 mg or less per day, more preferably 500 mg or less, and particularly preferably 200 mg or less.

[0026] The composition of the present invention may be appropriately designed so that the daily intake of oenothein B is the above-mentioned amount, and may be in a form in which the above-mentioned amount is taken in one dose or in a form in which the above-mentioned amount is taken in multiple doses. That is, for example, the composition may be stored in one container or in 2 to 4 multiple containers as a daily amount.

[0027] The packaging form of the composition of the present invention is not particularly limited and can be appropriately selected depending on the dosage form, etc., and examples thereof include blister packs such as PTPs; strip packaging; heat seals; aluminum pouches; film packaging using plastics, synthetic resins, etc.; glass containers such as vials; and plastic containers such as ampoules. EXAMPLES

[0028] The present invention will be described in more detail below with reference to examples. However, the present invention is not limited to these examples, and the present invention can take various forms as long as the object of the present invention can be achieved.

[0029] <Test> Evaluation of the effect of promoting uroplakin gene expression In order to verify that the composition of the present invention has the effect of improving frequent urination by improving overactive bladder, the effect of promoting the expression of the uroplakin (hereinafter abbreviated as "UPK") gene, a protein expressed in the urothelium of the ureter, bladder, etc., was evaluated.

[0030] [Test substance] In the examples, oenothein B (manufactured by SIGMA) was used. For comparison, quercetin (manufactured by SIGMA), kaempferol (manufactured by FUJIFILM Wako Pure Chemical Industries, Ltd.), gallic acid (manufactured by SIGMA), and ellagic acid (manufactured by FUJIFILM Wako Pure Chemical Industries, Ltd.) were used.

[0031] [Cell culture] As a model cell of bladder epithelial cells, T24 (manufactured by JCRB), which is a cell derived from bladder transitional cell carcinoma, was used and cultured in 10% FBS-MEM medium. The above cells were seeded in a 96-well plate at 100 μL / well so that the number of cells was 8000 cells / well, and precultured for 24 hours at 37°C in a 5% by volume CO2 incubator. After removing the medium from each well, 100 μL / well of medium containing the test substance was added and cultured for 24 hours. The medium used was 0.5% DMSO-10% FBS-MEM medium. The concentration of the test substance in the medium was as shown in Table 1.

[0032] [Table 1]

[0033] [Measurement of UPK gene expression level] After removing the medium from each well, RNA was extracted using RNeasy Mini Kit (QIAGEN) and quantitative real-time PCR was performed using One Step TB Green PrimeScript RT-PCR Kit II (Perfect Real Time) (Takara). The gene expression levels of UPK2 and UPK3A were measured using UPK2 primers (QIAGEN) and UPK3A primers (Eurofins Genomics). In addition, the gene expression level of ACTB was measured using ACTB primers (QIAGEN) as an endogenous control.

[0034] [Evaluation of UPK gene expression] The gene expression levels of UPK2 and UPK3A were calculated relative to the value in the control group (Comparative Example 1) taken as 1. The results are shown in FIGS.

[0035] [result] As shown in Figures 1 and 2, the group administered with oenothein B (Example 1) showed increased gene expression levels of UPK2 and UPK3A compared to the control group (Comparative Example 1). Furthermore, the group administered with oenothein B (Example 1) showed increased gene expression levels of UPK2 and UPK3A compared to the groups administered with quercetin (Reference Example 1), kaempferol (Reference Example 2), gallic acid (Reference Example 3), and ellagic acid (Reference Example 4). Therefore, since the composition of the present invention shows an excellent effect of promoting uroplakin gene expression, it is expected to have an effect of improving frequent urination by improving overactive bladder.

[0036] <Test> Evaluation of ROS production inhibitory effect In order to verify that the composition of the present invention has an effect of improving frequent urination by improving overactive bladder, the ROS production inhibitory effect was evaluated.

[0037] [Test substance] In the examples, onothein B used was onothein B (manufactured by SIGMA) and a hydroethanolic extract of willow herb containing onothein B. The content of onothein B in the hydroethanolic extract of willow herb was measured by HPLC.

[0038] [Cell culture] As a model cell of bladder epithelial cells, T24 (manufactured by JCRB), which is a cell derived from bladder transitional cell carcinoma, was used and cultured in 10% FBS-MEM medium. The above cells were seeded in a 96-well plate at 100 μL / well so that the number of cells was 10,000 cells / well, and precultured for 18 hours at 37°C in a 5% CO2 incubator. After removing the medium from each well, 100 μL / well of medium containing the test substance was added and cultured for 24 hours. The medium used was 0.5% DMSO-10% FBS-MEM medium. The final concentration of the test substance in the medium was as shown in Table 2.

[0039] [Table 2]

[0040] [Measurement of ROS production] After removing the medium from each well, the cells were washed twice with PBS, and 50 mM H2O2 prepared in the medium was added to give oxidative stress. In the normal group (Comparative Example 3), medium was added instead of 50 mM H2O2 prepared in the medium, and oxidative stress was not given. The cells were cultured for 3 hours in a 37°C, 5% by volume CO2 incubator. After removing the medium from each well, the cells were washed once with PBS, and 10 μM DCFH-DA reagent (manufactured by SIGMA) was added, and the cells were cultured for 30 minutes in a 37°C, 5% by volume CO2 incubator. After removing the medium from each well, the cells were washed twice with PBS, and the ROS production amount was measured by measuring the fluorescence intensity derived from DCF (excitation wavelength 485 nm, fluorescence wavelength 530 nm) using a plate reader (manufactured by Molecular Devices).

[0041] [Evaluation of ROS production] The amount of ROS produced was calculated as a relative value when the value of the control group (Comparative Example 2) was set at 100. The results are shown in FIG.

[0042] [result] As shown in FIG. 3, the group administered with oenothein B (Example 2) and the group administered with the hydroethanolic extract of willow herb (Example 3) showed suppression of ROS production compared to the control group (Comparative Example 2). In addition, the group administered with oenothein B (Example 2) and the group administered with the hydroethanolic extract of willow herb (Example 3) showed the same effect of suppressing ROS production. Since the group administered with oenothein B (Example 2) and the group administered with the hydroethanolic extract of willow herb (Example 3) were administered with the same dose of oenothein B, it is considered that oenothein B is the active ingredient in the hydroethanolic extract of willow herb that has the effect of suppressing ROS production. Therefore, since the composition of the present invention shows an excellent effect of suppressing ROS production, it is expected to have an effect of improving frequent urination by improving overactive bladder.

[0043] <Production Example> Based on the results of the Examples, Production Examples of the present invention are given below.

[0044] [Production Example 1-4: Granules] According to the formulation in Table 3, oenothein B and other raw materials were mixed, and then fluidized bed granulation was performed using a granulator to produce the granules described in Production Example 1-4. The granules described in Production Example 1-4 may be taken by dissolving 3 g per day in 100 ml of water or the like, or may be taken as is without dissolving. All of the granules in Production Examples 1-4 can improve overactive bladder and are effective in improving frequent urination.

[0045] [Table 3]

[0046] [Production Example 5-8: Tablets] According to the formulation in Table 4, oenothein B was mixed with other raw materials, and then tableted using a rotary tablet press to produce tablets (250 mg) of Production Example 5-8. One to two tablets of the tablets described in Production Example 5-8 may be taken per day with water or the like. All tablets of Production Example 5-8 can improve overactive bladder and are effective in improving frequent urination.

[0047] [Table 4]

[0048] [Production Example 9-12: Hard Capsule] According to the formulation in Table 5, oenothein B was mixed with other raw materials and encapsulated with a film containing gelatin or hydroxypropyl cellulose to produce hard capsules (contents: 300 mg). One to two capsules per day can be taken with water. All of the hard capsules in Manufacturing Examples 9 to 12 can improve overactive bladder and are effective in improving frequent urination.

[0049] [Table 5]

[0050] [Manufacturing example 13-16: PET beverage] According to the formula in Table 6, a liquid formulation containing oenothein B and other ingredients was mixed and packed into a PET container to produce a PET beverage (500 ml). One bottle should be consumed per day. All of the PET beverages in Manufacturing Examples 13-16 can improve overactive bladder and are effective in improving frequent urination.

[0051] [Table 6] [Industrial Applicability]

[0052] The composition of the present invention is industrially useful since it can improve frequent urination and can be used as a health food or the like.

Claims

1. A composition for improving frequent urination, comprising oenothein B as an active ingredient.

2. A composition for improving overactive bladder, comprising oenothein B as an active ingredient.

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