JP6550234B2 - TRPV4 activity inhibitor - Google Patents

Description

  The present invention relates to TRPV4 activity inhibitors and agents for preventing or improving overactive bladder.

  TRPV4 (Transient receptor potential cation channel subfamily V member 4) is one of the proteins constituting the temperature sensitive TRP channel. TRPV4 is expressed in a wide range of tissues such as kidney, lung, bladder, heart, skin, brain, gastrointestinal tract, and is considered to play different and broad physiological roles.

TRPV4 is particularly strongly expressed in the distal tubular epithelial cells of the kidney, and it has been suggested that the osmotic pressure and flow rate of urine can be sensed by the action of TRPV4 (see Non-patent Document 1). In addition, TRPV4 has been reported to be involved in bladder function (see Non-Patent Document 2). That is, TRPV4 exists in the bladder epithelial cells inside the bladder, and when urine accumulates and the bladder epithelial cells extend, calcium flows into the cells via the TRPV channel constituted by TRPV4 and the like. This influx of calcium releases ATP from the cells and the bladder bulges to the nerves.
In addition, it is reported that the effect of TRPV4 on the micturition interval is greater than that of other proteins that constitute TRP channels (see Non-Patent Document 3).
Therefore, inhibition of TRPV4 activity is expected to have a preventive or ameliorating effect on overactive bladder characterized by increased urgency and increased urination frequency.

So far, flavonoid compounds such as catechin are known to have an inhibitory action on TRPV4 activity (see Patent Document 1).
On the other hand, piperine is reported to be an agonist of TRPV1 and an agonist of vanilloid receptor (see Patent Documents 2 and 3). However, it is not known that piperine has an inhibitory action on TRPV4 activity and is useful for the prevention or improvement of overactive bladder.

JP 2014-24809 A International Publication No. 2011/060073 brochure WO 2008/040360 brochure

Tian W., Am. J. Physiol. Renal. Physiol., 2004, 287, p. F17-F24 Nilius B., Physiol. Rev., 2007, 87, p. 165-217 Gevaert T., J.A. Clin. Invest., 2007, 117, p. 3453-3462

An object of the present invention is to provide a TRPV4 activity inhibitor that inhibits TRPV4 activity.
Another object of the present invention is to provide a preventive or ameliorating agent for overactive bladder as a means of its administration utilizing its efficacy as a TRPV4 activity inhibitor.

  In view of the above problems, the present inventors diligently studied materials that inhibit TRPV4 activity. As a result, it was found that piperine has an effect of inhibiting TRPV4 activity. Thus, it has been found that this compound is useful for the prevention or improvement of overactive bladder. The present invention has been completed based on these findings.

The present invention relates to a TRPV4 activity inhibitor comprising piperine as an active ingredient.
The present invention also relates to an agent for preventing or improving overactive bladder, which comprises piperine as an active ingredient.

The TRPV4 activity inhibitor of the present invention can effectively inhibit TRPV4 activity.
The agent for preventing or improving overactive bladder of the present invention can also prevent or improve overactive bladder caused by activation of TRPV4 channel.

As used herein, “TRPV4” means “Transient receptor potential cation channel subfamily V member 4”. And this TRPV4 is a protein encoded by TRPV4 gene in humans.
TRPV4 is present in bladder epithelial cells inside the bladder. When urine accumulates and bladder epithelial cells expand, calcium is taken into cells via TRPV channels, whereby ATP is released from the cell surface and bladder swelling is transmitted to nerves. Therefore, by inhibiting the activity of TRPV4, overactive bladder can be prevented or ameliorated.

Here, "inhibition of TRPV4 activity" refers to suppressing the activity of the receptor TRPV4. Specifically, an activity expressed by binding a TRPV4 stimulant to TRPV4, for example, the ability to modulate ion flux (eg, ability to transport calcium ion, cation such as sodium ion from extracellular to intracellular, etc.) And means to suppress or inhibit the ability to control membrane potential (eg, the ability to generate current).
Further, in the present specification, "overactive bladder" is a symptom syndrome in which a feeling of urgency is essential, and usually refers to those with frequent urination and urge incontinence.

As used herein, "preventing" refers to preventing or delaying the onset of a disease or condition in an individual, or reducing the risk of developing an individual's disease or condition.
Moreover, in the present specification, “improvement” refers to amelioration of disease, symptom or condition, prevention or delay of deterioration of disease, symptom or condition, or reversal, prevention or delay of progression of disease, symptom or condition.
Furthermore, in the present specification, “non-therapeutic” is a concept that does not include medical practice, that is, treatment of the human body by treatment.

  The TRPV4 activity inhibitor of the present invention, and a preventive or ameliorating agent for overactive bladder are disclosed in piperine (1- [5- (1,3-benzodioxol-5-yl) -1-oxo-2 of the following structural formula , 4-pentadienyl] piperidine) as an active ingredient. In addition, piperine can be applied to inhibit TRPV4 activity and to prevent or improve overactive bladder.

  The compounds used in the present invention may be commercially available products or can be produced according to a conventional method (for example, G. Raman et al., Ind. Eng. Chem. Res., 2002, vol. 41). , P. 2966-2976). In addition, isomers such as geometric isomers, optical isomers, stereoisomers, etc. exist in the compound. Here, the compounds used in the present invention are not limited to the compounds of the above structural formula, but also include isomers of these compounds. Furthermore, the compound used in the present invention may be any isomer or a mixture of isomers.

As demonstrated in the examples below, piperine inhibits TRPV4 activity. Specifically, when a cell transformed with TRPV4 gene (TRPV4 expressing cell) is brought into contact with piperine in the presence of a TRPV4 stimulator (TRPV4 agonist, agonist), the intracellular cation amount by the TRPV4 stimulator And inhibit the activity of TRPV4. Thus, piperine is useful for the inhibition of TRPV4 activity.

  The form of the inhibitor of TRPV4 activity of the present invention and the agent for preventing or improving overactive bladder can be appropriately selected. For example, the active ingredient alone may be used as the TRPV4 activity inhibitor of the present invention, or as a preventive or ameliorating agent for overactive bladder. Alternatively, the TRPV4 activity inhibitor of the present invention containing the active ingredient and a pharmaceutically acceptable carrier, or an agent for preventing or improving overactive bladder may be used as a pharmaceutical composition. Alternatively, the cosmetic composition may contain the inhibitor of TRPV4 activity of the present invention, or the agent for preventing or improving overactive bladder.

  When preparing a pharmaceutical composition, it is usually prepared as a preparation containing the active ingredient and preferably a pharmaceutically acceptable carrier. A pharmaceutically acceptable carrier generally refers to an inert, non-toxic, solid, liquid, bulking agent, diluent or encapsulating material which does not react with the active ingredient, such as water And solvents such as ethanol, polyols (eg, propylene glycol, butylene glycol, glycerin, polyethylene glycol and the like), suitable mixtures thereof, solvents or dispersion media such as vegetable oils, and the like.

  The pharmaceutical composition may be orally, parenterally, for example, in the oral cavity, in the skin, subcutaneously, in the mucous membrane, intravenously, in artery, intramuscularly, intraperitoneally, vaginally, in the lung, Administered in the brain, in the eye, or in the nasal cavity. The oral administration preparations include tablets, granules, fine granules, powders, capsules, chewables, pellets, syrups, solutions, suspensions and inhalants. For parenteral administration, suppositories, holding enemas, drips, eye drops, nasal drops, pessaries, injections, mouthwashes, as well as ointments, creams, lotions, gels, controlled release patches And external preparations for skin such as patches. The pharmaceutical composition may be administered parenterally in the form of a sustained release subcutaneous implant, or in the form of a targeted delivery system (eg, monoclonal antibody, vector delivery, ion injection, polymer matrix, liposomes and microspheres) .

  The pharmaceutical composition may further contain conventional additives in the pharmaceutical field. Such additives include, for example, excipients, binders, disintegrants, lubricants, antioxidants, coloring agents, flavoring agents and the like, which can be used if necessary. In order to make a sustained release so as to be able to act for a long time, it can be coated with a known retarder or the like. As the excipient, for example, sodium carboxymethylcellulose, agar, light anhydrous silicic acid, gelatin, crystalline cellulose, sorbitol, talc, dextrin, starch, lactose, sucrose, glucose, magnesium aluminometasilicate, calcium hydrogen phosphate etc. It can be used. Examples of binders include gum arabic, sodium alginate, ethylcellulose, sodium caseinate, sodium carboxymethylcellulose, agar, purified water, gelatin, starch, tragacanth, lactose and the like. As the disintegrant, for example, carboxymethylcellulose, carboxymethylcellulose sodium, carboxymethylcellulose calcium, crystalline cellulose, starch, hydroxypropyl starch and the like can be mentioned. As the lubricant, for example, stearic acid, calcium stearate, magnesium stearate, talc, hydrogenated oil, sucrose fatty acid ester, waxes and the like can be mentioned. Examples of the antioxidant include tocopherol, gallic acid ester, dibutylhydroxytoluene (BHT), butylhydroxyanisole (BHA), ascorbic acid and the like. Other additives and drugs as required, such as antacid (sodium hydrogen carbonate, magnesium carbonate, precipitated calcium carbonate, synthetic hydrotalcite, etc.), gastric mucosa protective agent (synthetic aluminum silicate, sucralfate, sodium copper chlorophyllin etc.) ) May be added.

  When the cosmetic composition is prepared, the form can be appropriately selected, and solutions, emulsions, powders, water-oil two-layer systems, water-oil-powder three-layer systems, gels, tablets such as solids, aerosols, It can be in any form such as mist, capsule and sheet. In addition, the product form of the cosmetic composition is also optional, for example, cleanser, makeup remover, lotion, cosmetic solution, pack, milk, skin care cosmetic such as cream and sunscreen, foundation, makeup base, lipstick, eye Makeup cosmetics such as shadows, eyeliners, mascara, eyebrows, blushers and nail enamels, hair shampoos, hair rinses, hair conditioners, hair cosmetics such as hair dyes and hair restorers, soaps, body soaps, deodorant cosmetics and bath preparations And body cosmetics, oral cosmetics such as dentifrices and mouthwashes, and aromatic cosmetics such as perfumes. Also, the cosmetic may belong to either cosmetic or quasi-drug under the Japanese Pharmaceutical Affairs Law.

The cosmetic composition includes other components commonly used in cosmetics, quasi-drugs and pharmaceuticals, such as powder components, liquid fats and oils, solid fats and oils, waxes, hydrocarbons, higher fatty acids, higher alcohols, esters, silicones, anions Surfactant, Cationic surfactant, Amphoteric surfactant, Non-ionic surfactant, Moisturizer, Water-soluble polymer, Thickener, Coating agent, UV absorber, Sequestering agent, Lower alcohol, Polyhydric alcohol Sugar, amino acids, organic amines, polymer emulsions, pH adjusters, skin nutrients, vitamins, antioxidants, antioxidant aids, perfumes, water, etc., if necessary, and manufactured by a conventional method it can.
Examples of components that can be added to other cosmetic compositions include preservatives (eg, ethyl paraben, butyl paraben, etc.), anti-inflammatory agents (eg, glycyrrhizinic acid derivatives, glycyrrhetinic acid derivatives, salicylic acid derivatives, hinokitiol, zinc oxide, allantoin etc. ), Whitening agents (eg, ascorbic acid and derivatives thereof, placenta extract, yukinoshita extract, arbutin etc.), various extracts (eg, oubac, ouren, sikon, peony, cerebrum, birch, sage, loquat, carrot, aloe) , Zebra oasis, iris, grapes, yokinin, oats, lilies, saffron, sage, sage, oonilis, ononis, garlic, chili pepper, chili, seaweed etc., activators (eg royal jelly, photosensitizer, cholesterol derivative etc) , Blood circulation enhancer For example, nonyl acid warylamide, nicotinic acid benzyl ester, nicotinic acid β-butoxyethyl ester, capsaicin, zingerone, cantalistinqui, ictamol, tannic acid, α-borneol, nicotinic acid tocopherol, inositol hexanicotinate, cichrandelate, cinnarizine , Trazoline, acetylcholine, verapamil, cephalantin, γ-oryzanol etc., antiseborrhoeic agent (eg sulfur, thianthol etc.), anti-inflammatory agent (eg tranexamic acid, thiotaurine, hypotaurine etc) and bactericidal agent (eg triclosan, And cetyl pyridinium chloride, thymols, benzalkonium chloride and the like.

  The pharmaceutical composition and the cosmetic composition can be applied in the form of an oral composition, an external composition, an oral composition and the like.

  The TRPV4 activity inhibitor of the present invention and the preventive or ameliorating agent for overactive bladder can be used by being added to or formulated into foods, beverages, feeds, pet foods. Alternatively, food and drink labeled with the concept of treatment, prevention or amelioration of a disease or condition which can be treated, prevented or ameliorated by inhibition of TRPV4 activity, ie health food, functional food, food for sick persons and specific It can be used by adding to or blending in health food and the like. The above-mentioned health food, functional food, food for sick persons, food for specified health use, specifically, various kinds of fine granules, tablets, granules, powders, capsules, syrups, solutions, liquid foods, etc. It can be used as a pharmaceutical form. The food in the form of a preparation can be produced in the same manner as a pharmaceutical preparation, and the active ingredient and a food-acceptable carrier such as a suitable excipient (eg starch, processed starch, lactose, glucose, water etc.), etc. And can be manufactured using conventional means. Furthermore, liquid food compositions such as soups, juices, milk drinks, tea drinks, coffee drinks, cocoa drinks, jelly drinks, sports drinks, diet drinks, semi-solid food compositions such as puddings and yogurts, breads, The TRPV4 activity inhibitor of the present invention or the agent for preventing or improving overactive bladder is added to any noodles, cookies, chocolate, candy, confections such as gum, rice crackers, spreads such as sprinkles, butter, jam, etc. The food composition can be manufactured by mixing or blending.

The content of the active ingredient in the TRPV4 activity inhibitor of the present invention and the agent for preventing or improving overactive bladder can be appropriately determined.
For example, the content of the active ingredient is preferably 0.00001% by mass or more, more preferably 0.0001% by mass or more, and 2% by mass or less in the total amount of the TRPV4 activity inhibitor of the present invention and the agent for preventing or improving overactive bladder 0.2 mass% or less is more preferable, 0.00001 to 2 mass% is preferable, and 0.0001 to 0.2 mass% is more preferable.

  Subjects for administration or intake of the TRPV4 activity inhibitor of the present invention and the agent for preventing or improving overactive bladder are preferably warm-blooded vertebrates, more preferably mammals. In the present specification, mammals include, for example, humans and non-human mammals such as monkeys, mice, rats, rabbits, dogs, cats, cattle, horses, and pigs. The TRPV4 activity inhibitor of the present invention and the agent for preventing or improving overactive bladder are suitable for human administration.

  The compounds used in the present invention, the TRPV4 activity inhibitor of the present invention, and the agent for preventing or improving overactive bladder can be preferably applied to a subject who wants to prevent or improve overactive bladder. In addition, the compound used in the present invention, the TRPV4 activity inhibitor of the present invention, and the agent for preventing or improving overactive bladder can be preferably applied under conditions where overactive bladder is evoked. Furthermore, it is preferable to orally administer the compound used in the present invention, the TRPV4 activity inhibitor of the present invention, and the agent for preventing or improving overactive bladder.

In the method for inhibiting TRPV4 activity of the present invention and the method for preventing or improving overactive bladder, the effective amount of the active ingredient applied by administration or intake is the condition of the individual, body weight, sex, age, activity of material, administration Alternatively, it may be determined as appropriate according to the route of administration, administration or schedule of intake, formulation form or other factors. For example, the effective amount of the active ingredient is preferably 0.1 mg or more, more preferably 0.5 mg or more, preferably 20 mg or less, more preferably 5 mg or less, or preferably 0.1 to 20 mg per kg of body weight per day. Preferably it is 0.5-5 mg. The active ingredient may be taken / administered once to several times a day, or at any time and interval.
In addition, administration or intake of the active ingredient may be systemic administration or intake, or may be local administration or intake.

  With regard to the embodiments described above, the present invention further discloses the following TRPV4 activity inhibitors, agents for preventing or improving overactive bladder, uses, and methods.

A TRPV4 activity inhibitor or an agent for preventing or improving overactive bladder, comprising <1> piperine as an active ingredient.

<2> The TRPV4 activity inhibitor according to <1>, wherein the TRPV4 is human-derived TRPV4.
<3> The agent for preventing or improving overactive bladder according to <1>, which prevents or improves overactive bladder by inhibiting TRPV4 activity.
<4> The total content of the TRPV4 activity inhibitor or the agent for preventing or improving overactive bladder, the content of the active ingredient is 0.00001% by mass or more, preferably 0.0001% by mass or more, and 2% by mass or less, preferably The TRPV4 activity inhibitor according to any one of <1> to <3>, which is 0.2% by mass or less, or an agent for preventing or improving overactive bladder.

<5> Use of piperine as a TRPV4 activity inhibitor or a preventive or ameliorating agent for overactive bladder.
<6> Use of piperine for producing a TRPV4 activity inhibitor, or a preventive or ameliorating agent for overactive bladder.
A method of using <7> piperine as a TRPV4 activity inhibitor, or a preventive or ameliorating agent for overactive bladder.
<8> A method for inhibiting TRPV4 activity or a method for preventing or improving overactive bladder, which comprises applying piperine.
<9> The use or method according to any one of <5> to <8>, wherein the TRPV4 is human-derived TRPV4.
The use or the method as described in any one of said <5>-<9> which prevents or improves an overactive bladder by inhibiting <10> TRPV4 activity.
The method as described in any one of said <7>-<10> which applies <11> piperine to the human who desires prevention or improvement of an overactive bladder.
The method of any one of said <7>-<11> applied under the conditions with which <12> overactive bladder was evoked.
The content of piperine is 0.00001% by mass or more, preferably 0.0001% by mass or more, and 2% by mass or less, preferably 0.2% of the total amount of the TRPV 4 activity inhibitor or the agent for preventing or improving overactive bladder. The use or method according to any one of the above <5> to <12>, which is not more than mass%.

<14> Piperine used for a method for inhibiting TRPV4 activity or a method for preventing or improving overactive bladder.
<15> Use of piperine for the manufacture of a TRPV4 activity inhibitor, or a preventive or remedy for overactive bladder.
<16> Use of piperine for inhibition of TRPV4 activity, or non-therapeutic method for prevention or improvement of overactive bladder.
<17> The compound or use according to any one of <14> to <16>, wherein said TRPV4 is human-derived TRPV4.
The compound or use of any one of the above <14> to <17>, which prevents or improves overactive bladder by inhibiting <18> TRPV4 activity.
The compound or use of any one of the above <14> to <18>, wherein <19> piperine is applied to human in whom it is desired to prevent or improve overactive bladder.
The compound or use of any one of the above <14> to <19>, wherein <20> piperine is applied under the condition where overactive bladder is evoked.
The compound or use of any one of the above <14> to <20>, wherein <21> piperine is applied in the form of a pharmaceutical composition or a cosmetic composition.
The compound or use of any one of the above <14> to <20>, wherein <22> piperine is applied in the form of food, beverage, or feed.
The content of <23> piperine is 0.00001% by mass or more, preferably 0.0001% by mass or more, and 2% by mass or less, preferably 0.2% by mass or less, any one of the above <14> to <22> Use described in.

<24> A non-therapeutic method for inhibiting TRPV4 activity, or a non-therapeutic method for preventing or improving overactive bladder, which comprises applying an effective amount of piperine.
<25> The method according to <24>, wherein the TRPV4 is human-derived TRPV4.
The method as described in said <24> or <25> term which prevents or improves an overactive bladder by inhibiting <26> TRPV4 activity.
The method according to any one of the above <24> to <26>, wherein <27> piperine is applied to a human who desires to prevent or improve overactive bladder.
The method of any one of said <24>-<27> applied under the conditions with which <28> overactive bladder was evoked.
The effective amount of <29> piperine is 0.1 mg or more, preferably 0.5 mg or more, and preferably 20 mg or less, preferably 5 mg or less per kg of body weight per day, in the above <24> to <28> The method according to any one of the preceding claims.

  EXAMPLES The present invention will be described in more detail based on examples given below, but the invention is not meant to be limited by these.

Preparation human duodenal derived cells in Test Example 1 Human TRPV4 gene expression vector Human (HuTu-80 cells, American Type Culture Collection purchased from) obtained by reverse transcription of totalRNA extracted from cDNA as a template, published TRPV4 The polymerase chain reaction (PCR) was performed under the following conditions using a primer set consisting of oligonucleotides represented by the base sequences shown below, which were synthesized with reference to the gene sequence.

<Primer set>
Forward primer; 5'-CACCATGGCGGATTCCAGCGAAGGCCC-3 '(SEQ ID NO: 1)
Reverse primer; 5'-CTAGAGCGGGGCGTCATCAGTCC-3 '(SEQ ID NO: 2)

<PCR conditions>
a) PCR solution composition
cDNA (Template) 15 μL
5 × PrimeStar GXL Buffer 10 μL
dNTPs mixture (2.5mM) 4μL
1 μl PrimeStar GXL DNA Polymerase (Takara Bio)
Forward Primer (10 μM) 1 μL
Reverse Primer (10 μM) 1 μL
Water 18 μL
b) Temperature and cycle condition 95 ° C 2 min
↓
98 ° C 10sec 33 cycles
70 ° C 2 min

The obtained PCR product was purified using High Pure PCR Product Purification Kit (trade name, manufactured by Roche). A human TRPV4 gene expression vector was produced using the purified PCR product and pcDNA3.1 Directional TOPO Expression Kit (trade name, manufactured by Invitrogen Corporation).

Test Example 2 Preparation of Human TRPV4 Expressing Cells HEK293 cells (purchased from American Type Culture Collection) were cultured using DMEM / F12 medium (manufactured by Invitrogen) containing 10% fetal bovine serum. HEK 293 cells were seeded at 2 × 10 ⁵ cells / flask in a flask for T-25 cell culture. After 3 days of culture, the human TRPV4 gene expression vector (3 μg) prepared in Test Example 1 was transfected into cells using TransIT-293 (manufactured by Mirus) and cultured for 1 day. Separately, LacZ expression vector (3 μg) was transfected into cells using TransIT-293 (Mirus) as a control and cultured for 1 day. The cells are detached with Detachin (manufactured by Genlantis), and seeded at a cell density of 1.66 × 10 ⁴ cells / 90 μL / well in a 96-well Optical bottom plate (manufactured by Nunc) in DMEM / F12 medium containing 10% fetal bovine serum. The cells were further cultured for one day.

Example 1 Measurement of Intracellular Ca ²⁺ Influx Activity Measurement of intracellular Ca ²⁺ influx activity was performed using Calcium Kit II-fluo 4 (trade name, manufactured by DOJINDO). 90 μL / well of Loading buffer containing Fluo 4-AM was added to the cells prepared in Test Example 2 and incubated at 37 ° C. for 1 hour. Thereafter, the fluorescence intensity (excitation wavelength: 488 nm, fluorescence wavelength: 524 nm) was measured every two seconds using a fluorescence plate reader FDSS 3000 (trade name, manufactured by Hamamatsu Photonics K. K.) at 37 ° C. Thirty seconds after the start of measurement, TRPV4 agonist GSK1016790A (Sigma, 10 nM (final concentration 1 nM)), TRPV4 specific inhibitor HC067047 (Sigma, 100 μM (final concentration 10 μM)), and 20 μL / well of the sample solution (final concentration 10 times concentration) was added, and the change in fluorescence intensity was measured every 2 seconds up to 300 seconds.

<Sample>
GSK1016790A (Sigma, TRPV4 agonist) and HC067047 (Sigma, TRPV4 specific inhibitor) were dissolved in dimethyl sulfate (DMSO, DOJINDO) and used.
Piperine (manufactured by Sigma) was dissolved in a solvent (ethanol special grade, manufactured by Wako Pure Chemical Industries, Ltd.) and used.
Nutmeg ( Myristica fragrans ) extract used as a comparative example: Nutmeg Glycid SP-77543 (manufactured by San-Ei Gen FFI).

The TRPV4 activity rate (%) of the sample was calculated by the following equation.

TRPV4 activation rate (%) = (F0-FmeanC2) / (FmeanC1-FmeanC2) * 100


Fmean C1: Average value of fluorescence intensity of wells added with GSK1016790A (final concentration 10 μM)
Fmean C2: Average value of fluorescence intensity of wells added with only DMSO
F0: fluorescence intensity of the well to which the sample is added

<Test method / Result>
TRPV4 activity is evaluated at 50 to 100 seconds after the start of measurement, TRPV4 activation rate is 100% when TRPV4 agonist GSK1016790A (final concentration 1 nM) is added, and TRPV4 activity when DMSO as control is added Conversion rate was 0%.
The results are shown in Table 1.

The activation rate to the TRPV4 agonist GSK1016790A when piperine was added was assayed by Dunnet. As a result, as shown in Table 1, TRPV4 activity was significantly inhibited (p <0.01) in the presence of the TRPV4 agonist, regardless of the piperine concentration.
On the other hand, application of the nutmeg extract reported to contain alkaloid (Journal of Pain Research, 2013, vol. 6, p. 611-615) did not inhibit TRPV4 activity.

  As described above, application of piperine significantly inhibits TRPV4 activity. This indicates that piperine is effective in inhibiting TRPV4 activity. Furthermore, piperine having an action of inhibiting TRPV4 activity has been shown to be effective for the prevention or improvement of overactive bladder.

Claims (3)

A TRPV 4 activity inhibitor comprising piperine as an active ingredient (except for pharmaceutical compositions for preventing or treating urination disorder and health functional food, which comprises an extract of Indian long pepper ( Piper Longum L.) as an active ingredient) . A pharmaceutical agent for preventing or treating urination disorder, which comprises piperine as an active ingredient, an agent for preventing or improving urinary urgency associated with overactive bladder (but containing an extract of Indian long pepper ( Piper Longum L.) as an active ingredient) Chemical compositions and healthy functional foods). A food and drink composition for preventing or improving urinary urgency associated with overactive bladder, comprising piperine as an active ingredient (but containing, as an active ingredient, an extract of Indian long pepper ( Piper Longum L.) as an active ingredient, or Excluding therapeutic pharmaceutical compositions and health functional foods).