Bladder relaxant for women and oral composition containing same

Abstract

To provide a tonic bladder relieving agent for women that causes no adverse effect, and can effectively exert a bladder smooth muscle contraction suppressing and muscarine receptor blocking effect, and a composition for oral ingestion containing the relieving agent.SOLUTION: A tonic bladder relieving agent for women having extract from saw palmetto fruit as an active ingredient, and a composition for oral ingestion containing the tonic bladder relieving agent for women.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a bladder relaxant for women and an orally ingestible composition containing the same. [Background technology]

[0002] Saw palmetto fruit extract is known as an extract from the fruit of Serenoa repens, a low shrub belonging to the palm family that grows wild in the southeastern United States. This extract is standardized in the United States Pharmacopeia (USP: The United States Pharmacopeial Convention) and the European Pharmacopoeia (EP: European Pharmacopoeia), and extraction methods include solvent extraction with ethanol or a mixture of n-hexane and methylpentane, and supercritical carbon dioxide extraction. In Japanese health foods, the supercritical carbon dioxide extraction method is used.

[0003] The extract is standardized to contain at least 80% fatty acids, including at least 23% lauric acid, and at least 0.2% total sterols, including at least 0.1% β-sitosterol. Due to its α1-adrenergic receptor blocking and 5α-reductase inhibitory effects, it is used worldwide as a pharmaceutical and health food for the treatment of benign prostatic hyperplasia and lower urinary tract symptoms in men (see, for example, Non-Patent Document 1).

[0004] On the other hand, for women, there are studies on the effectiveness of a mixture of echinacea and saw palmetto fruit extract and therapeutic drugs for idiopathic megacystis (see, for example, Non-Patent Document 2 and Patent Document 1).

[0005] Furthermore, based on a study of the effectiveness of isosamidins derived from Peucedanum japonicum in women with overactive bladder, they have been commercialized as a supplement for urinary disorders in the form of a mixture with saw palmetto fruit extract (see, for example, Non-Patent Document 3 and Patent Documents 2 and 3).

[0006] Cucurbita pepo seed extract is also said to be effective for benign prostatic hyperplasia, and although there has been little research on it in women, it is also used as a supplement for female urinary disorders in combination with saw palmetto fruit extract (see, for example, Non-Patent Document 4 and Patent Documents 4 and 5).

[0007] Lower urinary tract symptoms are also referred to as urination problems, depending on the individual's subjective opinion, and consist of storage symptoms, urination symptoms, and post-urination symptoms, and include frequent urination, nocturia, urgency, urinary incontinence, weak urinary stream, straining urination, feeling of residual urine, difficulty urinating, and bladder pain. The bladder, which is involved in urination, is multiply innervated by three types of nerves: parasympathetic, sympathetic, and somatic nerves. In overactive bladder, urine tends to be released more easily, so suppression of parasympathetic excitation, which promotes urination by contracting the detrusor muscle, is necessary. Acetylcholine is involved in parasympathetic excitation, and anticholinergic drugs that suppress the action of acetylcholine are used in treatment. Another method uses agonists that stimulate β3-adrenergic receptors in the sympathetic nervous system, which act inversely to the parasympathetic nervous system (see, for example, Non-Patent Document 5). Furthermore, vanilloid receptors (transient receptor potential vanilloid subtype 1; TRPV1) are nociceptors present in peripheral sensory nerve endings that sense pain and temperature. In recent years, TRPV1 expression has been confirmed in the bladder, and it has been revealed that nerve excitation in sensory afferents is closely related to smooth muscle contraction and is involved in urinary function.

[0008] A study in Japan on the relationship between urinary disorders and health found that approximately 78% of men and women aged 60 or older have some form of lower urinary tract symptoms, and in some cases, no lower urinary tract dysfunction or other pathological conditions are recognized, or the nature of the symptoms does not lead to medical consultation or treatment, which can cause urination problems and a decline in quality of life (QOL) (see, for example, non-patent document 6). [Prior art documents] [Patent documents]

[0009] [Patent Document 1] Special Publication No. 2011-512371 [Patent Document 2] Japanese Patent Application Laid-Open No. 2015-083564 [Patent Document 3] Japanese Patent Application Laid-Open No. 2015-10068 [Patent Document 4] Japanese Patent Application Laid-Open No. 2012-46459 [Patent Document 5] Japanese Patent Application Laid-Open No. 2005-343809 [Non-patent literature]

[0010] [Non-Patent Document 1] Consumer Affairs Agency (Contractor: Japan Health and Nutrition Food Association, Public Interest Incorporated Foundation): Food Functionality Evaluation Model Project: Functionality Evaluation Survey Report: 141-150. (2012) [Non-patent document 2] Timmermans LM, et al,:Determination of the activity of extracts of Echinaceae and Sabal in the treatment of idiopathic megabladder in women:Acta urogica Belgica:58(2):43-59.(1990) [Non-patent document 3] Shizuo Yamada: Extension of healthy life expectancy and the complementary medical benefits of functional foods: Pharmaceutical Sciences: 77(5):264-269. [Non-patent document 4] National Institute of Biomedical Innovation, Health and Nutrition Safety and Efficacy Information for "Health Foods" [Non-patent document 5] Guidelines for the treatment of female lower urinary tract symptoms: Japan Council for Quality Health Care [Non-patent document 6] Homma, Y., Kakizaki, H., Goto, M., et al.: Epidemiological studies on urination: Journal of the Japanese Society of Urination: 14(2):266-277. (2003) Summary of the Invention [Problem to be solved by the invention]

[0011] The effectiveness and mechanism of saw palmetto fruit extract for treating benign prostatic hyperplasia and lower urinary tract symptoms in men have been clarified. However, although saw palmetto fruit extract has been used in combination with other ingredients for women, its effectiveness in treating lower urinary tract symptoms in women alone has not been clarified. This is thought to be because saw palmetto fruit extract was originally marketed as a treatment for male hormones and benign prostatic hyperplasia, and its effectiveness in women has not been proven. Furthermore, because saw palmetto fruit extract acts on male hormones, it may disrupt the hormonal balance of fetuses and infants, and therefore its use is discouraged in pregnant and nursing women. Based on these facts, it is inferred that the effectiveness of saw palmetto fruit extract alone for treating lower urinary tract symptoms in women has not been clarified.

[0012] In addition, anticholinergic drugs are used for the pharmacological treatment of overactive bladder associated with lower urinary tract symptoms in women, but anticholinergic adverse events such as nausea, constipation, and cognitive decline are major problems, especially in the elderly. In other words, there are no health foods for lower urinary tract symptoms in women that are free from side effects and have proven mechanisms and effectiveness.

[0013] For these reasons, there is a need for an improving agent for women that has no side effects, does not require a prescription, and is effective for relatively mild lower urinary tract symptoms, etc., in patients other than those who do not require treatment.

[0014] The present invention has been made in consideration of the above, and aims to provide a bladder relaxant for women that has no side effects and can effectively inhibit bladder smooth muscle contraction and block muscarinic receptors, and an orally ingestible composition containing the same. [Means for solving the problem]

[0015] In order to solve the above problems, the present inventors have measured urinary function using a female rat urinary disorder model, measured the contractile and test effects of isolated female rat bladder smooth muscle, and measured the binding activity of muscarinic receptors and β3 receptors in the female rat bladder, and have found that saw palmetto fruit extract has the effect of improving frequent urination and relaxing bladder tone during the urine storage period in female rats, and have discovered a blocking effect on muscarinic receptors due to the inhibitory effect on bladder smooth muscle and acetylcholine contraction, and an inhibitory effect on TRPV1, thereby completing the present invention. That is, the present invention is as follows.

[0016] [1] Saw palmetto fruit extract only A bladder relaxant for women containing the active ingredient and A bladder relaxant for women, wherein the active ingredient is a component that is effective in inhibiting bladder smooth muscle contraction and blocking muscarinic receptors, and a component that exhibits an inhibitory effect on vanilloid receptors. . [2] Taking the active ingredient in an amount of 58 mg or more per day as lauric acid can improve the number of urinations during the day. [1] A bladder relaxant for women as described. [3] [1] An orally ingestible composition comprising the bladder relaxant for women described in the above. [Effects of the Invention]

[0017] According to the present invention, it is possible to provide a bladder relaxant for women that has no side effects and can effectively inhibit bladder smooth muscle contraction and block muscarinic receptors, and an orally ingestible composition containing the same. This makes it possible to provide an orally ingestible composition that improves unpleasant lower urinary tract symptoms such as overactive bladder, frequent urination, and incontinence in women caused by aging, stress, obesity, etc. Furthermore, it provides an effective alleviating agent for relatively mild lower urinary tract symptoms that has no side effects, does not require a prescription, and does not require medical treatment, which can lead to improvements in the concerns and quality of life of many women. [Brief explanation of the drawings]

[0018] [Figure 1]FIG. 1 shows changes in intravesical pressure in Example 1, where pre shows the relationship between time and intravesical pressure after a single oral administration of excipient, and post shows the relationship between time and intravesical pressure after a single oral administration of SPE (160 mg / kg). [Figure 2] 1A and 1B are graphs showing the SPE concentration dependency in Example 2, in which FIG. 1A shows the relationship between SPE concentration and contraction of the isolated rat bladder due to low-frequency (0.1 Hz) transmural electrical stimulation (EFS), FIG. 1B shows the relationship between SPE concentration and contraction of the isolated rat bladder due to low-frequency (0.1 Hz) and high-frequency (1 Hz) transmural electrical stimulation (EFS), and FIG. 1C shows the relationship between SPE concentration and contraction of the isolated rat bladder due to acetylcholine (10 μM). [Figure 3] FIG. 10 is a diagram showing the relationship between the SPE concentration and the specific binding amounts of [ 3 H]NMS and [ 125 I]CYP in Example 3. [Figure 4] FIG. 1 shows the relationship between SPE concentration and specific binding of [ 3 H]Resiniferatoxin (RTX) to HEK293VR11 cells. DETAILED DESCRIPTION OF THE INVENTION

[0019] [Bladder tone relaxant] The bladder relaxant of the present invention contains saw palmetto extract (hereinafter sometimes referred to as "SPE"). only It is a bladder relaxant for women, whose active ingredient is

[0020] Here, the above-mentioned "active ingredient" is preferably an ingredient that is effective in inhibiting bladder smooth muscle contraction and blocking muscarinic receptors. For example, as a specific lower urinary tract symptom, it is an ingredient that is effective in improving the number of urinations during the day. Saw palmetto fruit extract was first discovered by the present inventors as such an active ingredient.

[0021] In addition, the active ingredient teeth,In particular, saw palmetto fruit extract alone can exert sufficient inhibitory effects on bladder smooth muscle contraction and muscarinic receptor blocking effects.

[0022] As described above, the bladder relaxant of the present invention contains an extract of saw palmetto fruit. only As long as it contains 80% or more fatty acids derived from saw palmetto fruit, the extraction method and final form (liquid, solid, powder) are not important.

[0023] The bladder relaxant of the present invention is a saw palmetto fruit extract. only It is used in oral compositions for women that have a bladder relaxation effect and contain the active ingredient as the main functional ingredient, and is widely used in, for example, over-the-counter (OTC) medicines and foods (foods for specified health uses, foods with functional claims, supplements, health foods, and clear foods). In particular, taking the active ingredient in an amount of 58 mg or more per day as lauric acid can improve the number of urinations during the day.

[0024] [Oral Ingestion Composition] The orally ingestible composition of the present invention contains the bladder relaxant for women of the present invention described above. That is, the orally ingestible composition of the present invention can also be said to be an orally ingestible composition for women. The oral composition of the present invention is not limited to any particular form as long as it is taken orally, but as mentioned above, it can be used as an OTC drug, food (foods for specified health uses, foods with functional claims, foods with nutrient functions, supplements, health foods, and clear foods), animal feed, supplements, etc.

[0025] There are no particular limitations on the oral intake form, but it is taken as tablets, capsules, powders, or drinks in OTC drugs, health foods, functional food products, supplements, and health foods. Because it contains a particularly large amount of fatty acids and has a unique unpleasant taste and odor, it can be formulated and provided alone or as a mixture in capsules or seamless capsules coated with gelatin, starch, carrageenan, etc. The oral composition of the present invention is preferably taken for 12 weeks or more so that the amount of lauric acid contained as a standardized component is 58 mg or more per day.

[0026] Furthermore, the composition for oral ingestion of the present invention may contain food ingredients or food additives such as preservatives, antioxidants, coloring agents, flavorings, seasonings, sweeteners, acidulants, glycerin fatty acid esters, emulsifiers such as soybean lecithin and beeswax, binders, strengthening agents, other excipients such as crystalline cellulose, lubricants, sugars, polysaccharides, starch, reducing sugars such as reduced malt, vitamins, minerals, proteins and amino acids such as ornithine, GABA, taurine, and theanine, carotenoids such as lycopene and lutein, polyphenols such as catechin, anthocyanin, soy isoflavone, and quercetin. It is also possible to blend in flavonoids or extracts containing them, lactic acid bacteria, fungi such as yeast, medium-chain fatty acids, plant-derived extract oils such as safflower oil, olive oil, and sea buckthorn oil, fatty acids and phospholipids such as DHA-refined fish oil, EPA-refined fish oil, and krill oil, dietary fiber such as dextrin, extracts and dried products of mushrooms, algae, animal and plant origins, or animal and plant secretions such as royal jelly and propolis, health food ingredients such as glucosamine, proteoglycan, chondroitin, collagen, chitin, and chitosan, and various pharmaceutical ingredients such as herbal medicines.It is also possible to clearly blend in foods.

[0027] It is also expected to be effective to combine and formulate materials that have β3 adrenergic receptor activity, which was not found in saw palmetto fruit extract. [Example]

[0028] Next, the present invention will be specifically explained with reference to examples, but the present invention is not limited to these examples.

[0029] Example 1: Effects on urinary function in a female rat urinary disorder model Female SD rats (10-15 weeks old) were given 0.5% acetic acid intravesically, and urinary function parameters (voiding interval, voiding volume, voiding frequency, maximum voiding pressure, voiding threshold pressure, and basal pressure) were measured by continuous measurement of intravesical pressure and voiding volume using cystometry. SPE (160 mg / kg) was administered orally to the rats once, and urinary function was measured before and after administration, and urinary parameters were calculated. In this example, the SPE used was SAW PALMETTO LIPIDIC STEROLIC EXTRACT, a product of EUROMED, ​​S.A.

[0030] Intravesical instillation of 0.5% acetic acid shortened the urination interval and increased the urination frequency. A single oral administration of SPE (160 mg / kg) to this frequent urination model rat resulted in an increase in the urination interval (62.6%) and a decrease in urination frequency (36.2%), both of which were statistically significant (Figure 1, Table 1). Furthermore, the volume of urine voided per void increased by 26.1%. While there was no change in maximum micturition pressure, there was a tendency for the micturition threshold pressure and basal pressure to decrease (17.5% and 28.4%, respectively). These results suggest that SPE increases bladder capacity and prolongs the urination interval. The decrease in basal pressure is thought to alleviate bladder tone during the urine collection phase.

[0031] [Table 1]

[0032] Each value in Table 1 above represents the mean ± standard error (n=7). An asterisk (*) indicates a significant difference from the value of the rats administered the vehicle ( * P<0.05).

[0033] (Example 2) Contraction and relaxation of bladder smooth muscle isolated from female rats Bladders were removed from female rats (12-14 weeks old) under deep anesthesia with pentobarbital, cut longitudinally into four quarters, and then suspended in a Magnus tube filled with Krebs-Henseleit solution under a resting tension of 0.3 g. In this example, the SPE used was SPE-SC, a product of BGG Japan.

[0034] Transmural electrical field stimulation (EFS) was performed by suspending the specimen between a ring-shaped anode and a rod-shaped cathode at its center. The stimulation conditions were as follows: frequency 0.1 and 1.0 Hz, duration 0.5 msec, and voltage 10 V. A 1.0 Hz EFS was applied for 10 s at 20 min intervals (Model SEN3301, Nihon Kohden). SPE was dissolved in DMSO (dimethyl sulfoxide) (100 mg / mL). In EFS experiments, SPE or vehicle (DMSO) was administered cumulatively under 0.1 Hz EFS stimulation. For 1.0 Hz EFS stimulation, SPE or DMSO was pretreated for 10 min before EFS administration. In ACh stimulation experiments, 10 μM ACh was administered at 10 min intervals, followed by 10 min of SPE or DMSO pretreatment.

[0035] The contractile response to each stimulus was recorded via a tension transducer (Model t-7, NEC Sanei) and an amplifier (Model AS1202, NEC Sanei) (Model 8K21, NEC Sanei). Data were calculated based on the tension change induced by 0.1 Hz EFS stimulation or 10 μM ACh without SPE or DMSO treatment (100%).

[0036] SPE had no effect on contractions of isolated rat bladders induced by low-frequency (0.1 Hz) transmural electrical stimulation at a concentration of 0.01 mg / ml, but significantly inhibited them at 0.1 and 1.0 mg / ml (16.0% and 32.4%, respectively) (Fig. 2A and B). Similarly, it significantly inhibited bladder contractions induced by high-frequency (1 Hz) transmural electrical stimulation (74.2%). Furthermore, SPE (0.01-1.0 mg / ml) inhibited bladder contraction induced by acetylcholine (10 μM) in a concentration-dependent manner (Fig. 2C). The inhibitory effect of 1.0 mg / ml SPE (30.3%) was significant.

[0037] Example 3: Effects of muscarinic receptor and β3 receptor binding activity in the urinary bladder of female rats Receptor preparations were prepared from the homogenate of the excised bladder of female rats. Binding activity to muscarinic receptors and β3 receptors was evaluated using [ 3 H]N-methylscopolamine ([ 3 H]NMS) and [ 125 I]cyanopindolol([ 125 The radioreceptor assay was performed using [I]CYP. In the presence of various concentrations of SPE, the receptor preparation was [ 3 H]NMS (0.10 nM) or [ 125 The cells were incubated with [I]CYP (0.12 nM), and the specific binding amount of each labeled ligand was measured by high-speed suction filtration using a Brandel cell harvester. In this example, the SPE used was SPE-SC, a product of BGG Japan.

[0038] In cells expressing muscarinic receptor subtypes (M1-M5), 3 The effect of SPE on the specific binding of [H]NMS was investigated. In the presence and absence of atropine (1 μM) 3 The difference in the amount of H]NMS bound to the muscarinic receptor was defined as the specific binding. SPE (10-300 μg / ml) was 3 The specific binding of [H]NMS was inhibited in a concentration-dependent manner (black circle plot in Figure 3). The 50% inhibitory concentration (IC 50 The value was 40.9±5.6 μg / ml. On the other hand, the values ​​measured in the presence and absence of 10 μM propranolol were 125 SPE (10-300 μg / ml) showed almost no inhibitory effect on the specific binding of CYP, indicating that SPE binds to muscarinic receptors in the rat bladder but not to β-receptors (open circles in Figure 3).

[0039] SPE (3-100 μg / ml) inhibited the muscarinic receptor subtype M1-M5 expression in cells [ 3 The IC20 calculated from the concentration-dependent inhibition of the specific binding of [H]NMS. 50 From Table 2, SPE tended to show higher binding affinity to M2, M3, and M4 subtypes compared to M1 and M5 subtypes.

[0040] [Table 2]

[0041] Table 2 shows the IC50 values ​​for the inhibition of specific binding of [3H]NMS to the membranes of CHO-K1 cell lines expressing human muscarinic receptor subtypes M1 to M5 by saw palmetto extract. Each value represents the mean ± standard error (n = 5).

[0042] SPE significantly improved urinary frequency in female rats with acetic acid-induced urinary frequency. It also relaxed bladder tone during the urine collection period. It significantly inhibited contractions of the bladder smooth muscle induced by transmural electrical stimulation and acetylcholine in female rats. This effect was thought to be partly mediated by blocking muscarinic receptors.

[0043] In addition, in a 28-day repeated oral administration toxicity test using female rats, no deaths were observed at a dose of 1000 mg / kg / day, and no changes that could be considered to be the toxic effects of saw palmetto fruit extract were observed in general condition, FOB, body weight, intake amount, hematological tests, blood biochemistry tests, estradiol concentration, estrous cycle observation, urinalysis, organ weight, necropsy, or histopathological tests. In this example, the SPE used was SAW PALMETTO LIPIDIC STEROLIC EXTRACT, a product of EUROMED, ​​S.A.

[0044] (Example 4) SPE concentration and [ 3Relationship between specific binding of [H]Resiniferatoxin (RTX) and human clinical trials HEK293VR11 cells, in which rat TRPV1 was stably expressed, were used. HEK293VR11 cells (1.8×10 7 The cells were seeded on a 150 mm cell culture dish and cultured for 3-4 days. 2+ After washing with free PBS, the cells were treated with 0.5 mM EDTA-containing Ca 2+ The cells were detached from the dish with 10 mL of free PBS and collected. The collected cells were washed once with 10 mL of Buffer A (10 mM HEPES, 5 mM KCl, 5.8 mM NaCl, 2 mM MgCl2, 0.75 mM CaCl2, 12 mM glucose, 137 mM sucrose; pH 7.4) and then diluted to 5.0 × 10 4 The mixture was diluted with Buffer A to a concentration of 100 cells / mL and used as a receptor sample.

[0045] Measurement of TRPV1 3 The radioreceptor assay was performed using a radioactive RT-PCR assay with 3H-RTX as a labeled ligand. The receptor preparation was incubated in an incubation buffer (Buffer A supplement with 0.25 mg / mL BSA; pH 7.4) with 1000 kJ / mL of [ 3 The cells were incubated with [H]RTX (0.0625-0.5 nM) at 37°C for 60 minutes. After incubation, the cells were left on ice for 5 minutes, and then 50 μL of 2 mg / mL α1-acid glycoprotein was added and incubated on ice for an additional 10 minutes. The reaction mixture was then rapidly filtered onto glass fiber filters (Whatmann GF / B) using a Cell Harvester for Receptor Binding Assay (Brandel, USA). The filters were immediately washed with 50 mM NaCl. + / K +The filter paper was washed twice with 3 mL of phosphate buffer (pH 7.5). The filter paper was placed in a vial, and toluene scintillator (2,5-diphenyloxazole: 5 g, 1,4-Bis(5-phenyl-2-oxazol)benzene: 0.1 g, Triton X-100: 333 mL, toluene: 666 mL) was added. After leaving the filter paper at room temperature for approximately 6 hours, the radioactivity was measured using a liquid scintillation counter. In this case, 1 μM RTX was used as a displacer. The radioactivity measured in the absence of displacer was defined as total binding, and the radioactivity measured in the presence of displacer was defined as nonspecific binding. The difference between the two was defined as specific binding to the receptor. All measurements were performed in duplicate.

[0046] Calculation of receptor binding parameters Using HEK293VR11 cells, the effects of various drugs on HEK293VR11 cells were investigated. 3 The specific binding of H]RTX (0.13 nM) was measured, and the [ 3 The drug concentration that inhibits the specific binding of [H]RTX by 50%, i.e., IC 50 The inhibition constant (K i ) was calculated.

[0047]

number

[0048] SPE was tested at concentrations of 3, 10, 30, 100, and 300 μg / mL on HEK293VR11 cells. 3 The specific binding of [H]Resiniferatoxin (RTX) was inhibited in a concentration-dependent manner (Figure 4).

[0049] Seventy-six women aged 50 or older with relatively mild lower urinary tract symptoms, excluding patients who did not require a prescription or treatment, were given one test food per day for 12 weeks. The test food was a gelatin capsule containing saw palmetto fruit extract at a concentration of 58 mg or more as lauric acid, and the placebo food contained the same amount of glycerin fatty acid ester instead of saw palmetto fruit extract. The test food group showed a significant improvement in the number of urinations from waking up in the morning until going to bed (number of urinations during the day) compared to the placebo food group. No serious illnesses or other side effects were reported in this study. In this example, the SPE used was SPE-SC, a product of BGG Japan.

Claims

1. It is a bladder relaxant for women, whose only active ingredient is saw palmetto fruit extract. A bladder relaxant for women, wherein the active ingredient is a component that is effective in inhibiting bladder smooth muscle contraction and blocking muscarinic receptors, and a component that exhibits an inhibitory effect on vanilloid receptors.

2. 2. The bladder relaxant for women according to claim 1, which can improve the number of urinations during the day by taking the active ingredient in an amount of 58 mg or more per day in terms of lauric acid.

3. An oral composition for women comprising the bladder relaxant for women according to claim 1.

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