JP2017052747A - Uroplakin expression-promoting agents - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide Uroplakin expression promoting agents that accelerate the expression of Uroplakin in bladder epithelial cells as well as agents for prevention or improvement of vesicoureteral reflux as a means for administration thereof.SOLUTION: The invention provides a Uroplakin expression promoting agent comprising at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid and a salt thereof and cumaric acid and a salt thereof as an active component. The invention also provides agents for preventing or improving vesicoureteral reflux.SELECTED DRAWING: None

Description

  The present invention relates to an uroplakin expression promoter.

  Cystoureteral reflux disease is a phenomenon in which urine collected in the bladder flows back to the ureter or renal pelvis / renal parenchyma due to the structure or abnormality of the vesicoureteral transition. It is the most common cause. In fact, 26-50% of children's urinary tract infections were found to have vesicoureteral reflux, 70% at 1 year of age, 40-50% at 1-2 years, and 5.2% at adults Frequency (Non-Patent Document 1). The presence of reflux can cause pyelonephritis due to ascending urinary tract infection, which can lead to renal failure due to repetition. Also, urine that has flowed back into the upper urinary tract during urination falls into the bladder after urination and becomes residual urine, so that bacteria grow and cystitis develops, causing bladder pain.

  Uroplakin (hereinafter referred to as UPK) in the present specification is a membrane protein specifically expressed in the urothelium. There are four types of UPKs known so far, UPK1A, UPK1B, UPK2 and UPK3A, and one of the causes of vesicoureteral reflux is suggested to involve UPK3A and UPK2. It has been reported that UPK3A and UPK2 knockout mice actually develop vesicoureteral reflux disease (see Non-Patent Document 2). In addition, UPK1A is known to form a heterodimer with UPK2 and UPK1B with UPK3A, respectively. In a UPK2 or UPK3A knockout mouse exhibiting vesicoureteral reflux, It has been reported that the expression level of UPK1A and UPK1B in the apical membrane decreases (see Non-Patent Document 3).

  In the treatment of vesicoureteral reflux disease, conservative therapy that waits for spontaneous disappearance is taken in mild cases, but when it does not disappear, reflux prevention is performed. However, surgical treatment is highly invasive, and a non-invasive method for preventing or improving vesicoureteral reflux is desired.

NEW Urology, 2007, 2nd revised edition, p. 135 Journal of Cell Biology, 2000, vol. 151, p. 961-971 Journal of Cell Biology, 2004, vol. 167, p. 1195-1204

The anti-reflux technique performed in the treatment of vesicoureteral reflux that does not disappear spontaneously is highly invasive, and there is no non-invasive and effective treatment.
On the other hand, if the decrease in the epithelial barrier function of the urinary bladder considered to be a cause of vesicoureteral reflux can be suppressed, it can be a causative therapy for vesicoureteral reflux.

An object of the present invention is to provide an agent for preventing or improving vesicoureteral reflux.
Moreover, this invention makes it a subject to provide the food-drinks composition for prevention or improvement of vesicoureteral reflux disease.
Another object of the present invention is to provide a UPK expression promoter that promotes the expression of UPK in bladder epithelial cells.
Another object of the present invention is to provide a food and drink composition for promoting UPK expression that promotes the expression of UPK in bladder epithelial cells.

Another object of the present invention is to provide a non-therapeutic prevention or improvement method for preventing or improving vesicoureteral reflux disease.
Another object of the present invention is to provide a UPK expression promoting method for promoting UPK expression in bladder epithelial cells.

In view of the above problems, the present inventors have conducted intensive studies on substances that can suppress the decrease in epithelial barrier function of the bladder that causes vesicoureteral reflux disease. As a result, it was found that rosmarinic acid, ferulic acid, and coumaric acid have an action of promoting the expression of the UPK gene and are useful for the prevention or improvement of vesicoureteral reflux disease.
The present invention has been completed based on these findings.

The present invention relates to a preventive or ameliorating agent for vesicoureteral reflux, comprising as an active ingredient at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof. About.
The present invention also relates to prevention or prevention of vesicoureteral reflux, comprising as an active ingredient at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof. It is related with the food-drinks composition for improvement.
The present invention also relates to a UPK expression promoter comprising as an active ingredient at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof.
The present invention also relates to a UPK expression promoting food or drink composition comprising as an active ingredient at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof. About.

The present invention also provides a non-therapeutic vesicoureteral reflux disease in which at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof is administered or ingested. Relates to a method for preventing or improving
The present invention also relates to a non-therapeutic method for promoting UPK expression by administering or ingesting at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof. .

The agent for preventing or improving vesicoureteral reflux and the food or beverage composition for preventing or improving vesicoureteral reflux can prevent or improve vesicoureteral reflux.
Moreover, the UPK expression promoting agent and the food / beverage product composition for promoting UPK expression of the present invention can promote the expression of UPK in bladder epithelial cells.

As used herein, “prevention” refers to preventing or delaying the onset of a disease or symptom in an individual, or reducing the risk of developing an individual's disease or symptom.
In the present specification, “improvement” refers to improvement 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.

The UPK expression promoter of the present invention is effective for promoting the expression of any of UPK1A, UPK1B, UPK2 and UPK3A. In particular, the UPK expression promoter of the present invention is effective for promoting the expression of the genes encoding the various UPKs.
The amino acid sequence information of human UPK1A and the base sequence information of the gene encoding it (hereinafter also referred to as “UPK1A gene”) are NCBI Reference Sequence: NP_001268372.1, NM_001281443.1, NP_008931.1, NM_007000. It is registered as 3 and is available from NCBI.
The amino acid sequence information of human UPK1B and the base sequence information of the gene encoding it (hereinafter also referred to as “UPK1B gene”) are registered as NCBI Reference Sequences: NP_008883.2 and NM_006952.3, respectively. It is available.
The amino acid sequence information of human UPK2 and the base sequence information of the gene encoding it (hereinafter also referred to as “UPK2 gene”) are registered as NCBI Reference Sequences: NP_006751.1 and NM_006760.3, respectively. More available.
The amino acid sequence information of human UPK3A and the base sequence information of the gene encoding it (hereinafter also referred to as “UPK3A gene”) are NCBI Reference Sequence: NP_001161046.1, NM_001167574.1, NP_008884.1, NM_006953. It is registered as 3 and is available from NCBI.
In addition, these isoforms, variants, or homologs can be targeted for expression promotion of the present invention.

The preventive or ameliorating agent for vesicoureteral reflux of the present invention (hereinafter also referred to as “preventing or improving agent of the present invention”), and the UPK expression promoter include rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumarin. The active ingredient is at least one compound selected from the group consisting of acids or salts thereof. The food / beverage composition for preventing or improving vesicoureteral reflux and the food / beverage composition for promoting UPK expression of the present invention comprise rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof. It contains at least one compound selected from the group as an active ingredient.
In the present invention, rosmarinic acid, ferulic acid and coumaric acid may each be used in the form of a salt. The salt is not particularly limited as long as it is a pharmaceutically acceptable salt, for example, a salt with an alkali metal such as sodium and potassium, a salt with an alkaline earth metal such as calcium and magnesium, ammonium and triethylamine, etc. And salts with amines. In the present invention, the salt is preferably a salt with an alkali metal or an alkaline earth metal, more preferably a sodium salt or a calcium salt, and even more preferably a sodium salt.
The preventive or ameliorating agent, UPK expression promoting agent, food / beverage composition for preventing or improving vesicoureteral reflux, and the food / beverage composition for promoting UPK expression include, as active ingredients, rosmarinic acid or a salt thereof, ferla Any one compound of an acid or a salt thereof and coumaric acid or a salt thereof may be used alone, or two or more compounds may be mixed and used.

There is no particular limitation on the production method of rosmarinic acid or its salt, ferulic acid or its salt, and coumaric acid or its salt, and it can be obtained by ordinary organic chemical synthesis. Alternatively, it can be obtained by extraction or purification from a natural product-derived material. Moreover, what is marketed as a reagent can also be used.
Rosmarinic acid or a salt thereof can be produced, for example, by suspension culture of Lamiaceae plant cells with reference to the method described in JP-B-1-29558.
Ferulic acid or a salt thereof can be produced, for example, by treating a plant material with an enzyme preparation with reference to the method described in JP-A-2014-57529.
Coumaric acid or a salt thereof can be produced using, for example, a coumaric acid-producing transformed microorganism with glucose as a carbon source with reference to the method described in JP-A No. 2014-117272.

As described above, it has been reported that UPK3A or UPK2 knockout mice develop vesicoureteral reflux disease. In addition, UPK1A is known to form a heterodimer with UPK2 and UPK1B with UPK3A, respectively. In a UPK2 or UPK3A knockout mouse exhibiting vesicoureteral reflux, It has been reported that the expression level of UPK1A and UPK1B in the apical membrane decreases.
From these reports, it is considered that urinary bladder reflux can be prevented or improved by promoting the expression of UPK.

As shown in the examples described later, rosmarinic acid, ferulic acid, and coumaric acid have the effect of promoting UPK gene expression in bladder epithelial cells. Rosmarinic acid is metabolized in vivo to ferulic acid and coumaric acid (Eur. J Nutr., 2005, vol44, p.1-9, Life Science, 2004, vol75, p.165-178) . Therefore, these compounds can be used to promote UPK expression and can be used to prevent or ameliorate vesicoureteral reflux.
The use may be a therapeutic use (ie medical practice) or a non-therapeutic use (non-medical practice). Moreover, the subject of the use can be a human, a non-human animal, or a specimen derived therefrom. The “non-therapeutic” is a concept that does not include a medical action, that is, a treatment action on the human body by treatment.

The preventive or ameliorating agent of the present invention is one specific embodiment of the above-described use, and can be applied to both therapeutic uses (medical uses) and non-therapeutic uses (non-medical uses). Specifically, the preventive or improving agent of the present invention can be added to various foods, foods, feeds, pet foods, etc., in addition to being usable as pharmaceuticals, quasi drugs and the like.
Further, the UPK expression promoter is one of the specific modes of use described above, and can be applied to both therapeutic uses (medical uses) and non-therapeutic uses (non-medical uses). Specifically, it can be used as a pharmaceutical product, quasi-drug, and the like, and a UPK expression promoter can be added to various foods, foods, feeds, pet foods, and the like.
The preventive or ameliorating agent and UPK expression promoting agent of the present invention can be applied to humans and animals in liquid, solid, emulsion, paste, gel, powder (powder), granule, pellet, stick, etc. Various dosage forms that can be applied can be taken.
In addition, the preventive or ameliorating agent and UPK expression promoter of the present invention may be composed only of the above-mentioned compounds that are active ingredients, or contain other ingredients within a range that does not affect the effect. There may be. Examples of the other components include the following additives.

When the preventive or ameliorating agent of the present invention and the UPK expression promoter are applied to pharmaceuticals and quasi-drugs, they can be prepared into various dosage forms by containing an effective amount of the compound and, if necessary, additives. . For example, as an oral medicine such as tablets, coated tablets, capsules, granules, powders, syrups, enteric solvents, troches, drinks, etc., or non-injectables such as injections, suppositories, transdermal absorption agents, external preparations, etc. It can be prepared as an oral medicine. Of these forms, the preferred form is oral medicine.
What is necessary is just to manufacture in accordance with a conventional method using an additive, in preparing in various dosage forms. As the additive, a commonly used additive can be used. Examples of additives include pharmaceutically acceptable excipients, liquid carriers, oily carriers, stabilizers, wetting agents, emulsifiers, binders, isotonic agents, disintegrants, lubricants, bulking agents, Surfactant, dispersant, suspending agent, diluent, osmotic pressure adjusting agent, pH adjusting agent, preservative, antioxidant, coloring agent, ultraviolet absorber, humectant, thickener, brightener, buffering agent, Preservatives, flavoring agents, fragrances, film agents, flavoring agents, bacteria inhibitors and the like can be mentioned.

When the preventive or ameliorating agent of the present invention, UPK expression promoter, or the above-mentioned active ingredient is blended and applied to foods and drinks, feeds, pet foods, etc., and is used as food / beverage product compositions, feed compositions, pet food compositions, etc. It can be provided by being formed into a form suitable for food or beverage, for example, granular, granular, tablet, capsule, paste or the like. Furthermore, in addition to general food and drink, the food and drink composition is based on the concept of prevention or improvement of vesicoureteral reflux disease or promotion of UPK expression, and if necessary, beauty food, food for the sick, It can be set as the food-drinks composition in the form of functional food-drinks, such as a nutritive functional food, the food for specified health, or a functional display food.
Examples of blending into foods and beverages include wheat flour processed foods, rice processed foods, confectionery, beverages, dairy products, seasonings, meat storage processed foods, processed fishery products, cooking oils and the like. In addition, tablets, tablets, and other tablet foods, concentrated liquid foods, natural liquid foods, semi-digested nutritional foods, ingredient nutritional foods, drink nutritional foods, etc. Good.
Examples of the feed composition include small animal feed used for rabbits, rats, mice and the like, pet food used for dogs, cats, small birds, squirrels and the like.
These food and beverage composition, feed composition, pet food composition and the like contain the preventive or ameliorating agent of the present invention, the UPK expression promoter of the present invention, or the above-mentioned active ingredient, and food ingredients such as, for example, Sweeteners, coloring agents, antioxidants, vitamins, fragrances, minerals and other additives, proteins, lipids, carbohydrates, carbohydrates, dietary fibers, etc. can be appropriately combined and prepared according to conventional methods.

The compounding amount or content of the active ingredient in the preventive or ameliorating agent, UPK expression promoter, and food / beverage product composition of the present invention can be appropriately determined depending on the form of use.
For example, in the case of oral solid preparations such as tablets, coated tablets, granules, powders, capsules, and oral liquid preparations such as oral liquids and syrups, the solid content concentration (solid content conversion) is 0.01% by mass. The above is preferable, 0.1 mass% or more is more preferable, 1 mass% or more is further preferable, 100 mass% or less is preferable, 90 mass% or less is more preferable, and 70 mass% or less is further preferable. Or 0.01-100 mass% is preferable, 0.1-90 mass% is more preferable, 1-70 mass% is further more preferable.
When the preventive or ameliorating agent of the present invention and the UPK expression promoter are blended in foods and drinks, pet foods, etc., the blending amount or content of the active ingredient is preferably 0.001% by mass or more as a solid content concentration, 0 0.01 mass% or more is more preferable, 0.1 mass% or more is more preferable, 50 mass% or less is preferable, 10 mass% or less is more preferable, and 3 mass% or less is further more preferable. Or 0.001-50 mass% is preferable, 0.01-10 mass% is more preferable, 0.1-3 mass% is further more preferable.

  Administration or intake of the preventive or ameliorating agent, UPK expression promoting agent, and food / beverage product composition of the present invention can be appropriately selected and determined according to the individual's condition, body weight, sex, age, or other factors. For example, the daily administration or intake of an adult (60 kg) is preferably 0.01 mg or more, more preferably 1 mg or more, still more preferably 10 mg or more, more preferably 100 mg or more, in terms of solid content of the compound as the active ingredient. Is more preferably 80000 mg or less, more preferably 40000 mg or less, further preferably 15000 mg or less, and even more preferably 5000 mg or less. Or 0.01-80000 mg is preferable, 1-40000 mg is more preferable, 10-15000 mg is further more preferable, and 100-5000 mg is still more preferable. In addition, the preventive or ameliorating agent, UPK expression promoting agent, and food / beverage product composition of the present invention can be taken or administered once a day to several times a day, or at an arbitrary period and interval.

  Although it does not specifically limit as an ingestion or administration object of the said pharmaceutical, a quasi-drug, or a food-drinks composition, The human and mammal other than a human aiming at prevention, improvement, and treatment of vesicoureteral reflux disease are preferable. In addition, the subject of ingestion or administration includes humans and non-human mammals with symptoms of vesicoureteral reflux disease, humans and non-human mammals that may be at risk thereof, and humans who are expected to prevent the disease / symptoms. And mammals other than humans are also included.

  The present invention further discloses the following preventive or ameliorating agent, UPK expression promoter, food / beverage product composition, and method for the above-described embodiments.

<1> An agent for preventing or improving vesicoureteral reflux, comprising as an active ingredient at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof.
<2> For prevention or improvement of vesicoureteral reflux, comprising as an active ingredient at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof Food and beverage composition.
<3> A UPK expression promoter comprising as an active ingredient at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof.
<4> A UPK expression promoting food or drink composition comprising as an active ingredient at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof.

<5> The UPK expression promoter according to <3> or <4>, or the UPK expression promoting food or drink, wherein the UPK is at least one selected from the group consisting of UPK1A, UPK1B, UPK2 and UPK3A Product composition.
<6> The content or content of the active ingredient relative to the total amount of the agent or food / beverage product composition is 0.01% by mass or more, preferably 0.1% by mass or more, more preferably 1% by mass in terms of solid matter. The prophylactic or ameliorating agent for vesicoureteral reflux according to any one of <1> to <5>, which is 100% by mass or less, preferably 90% by mass or less, more preferably 70% by mass or less. , UPK expression promoter, or food and beverage composition.
<7> The compounding amount or content of the active ingredient relative to the total amount of the agent or the food / beverage product composition is 0.001% by mass or more, preferably 0.01% by mass or more, more preferably 0.1% in terms of solid matter. Prevention of vesicoureteral reflux according to any one of the above <1> to <5>, which is not less than 50% by mass and not more than 50% by mass, preferably not more than 10% by mass, more preferably not more than 3% by mass. An improver, UPK expression promoter, or food or beverage composition.
<8> Prevention or amelioration of vesicoureteral reflux is due to promotion of UPK expression, preferably promotion of UPK expression in bladder epithelial cells, more preferably promotion of UPK gene expression in bladder epithelial cells. <1>, <2>, <6> and <7> The preventive or ameliorating agent for vesicoureteral reflux or the food or drink composition for preventing or improving vesicoureteral reflux object.

<9> Non-therapeutic prevention of vesicoureteral reflux disease by administering or ingesting at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof Or improvement method.
<10> Prevention or amelioration of vesicoureteral reflux is due to promotion of UPK expression, preferably promotion of UPK expression in bladder epithelial cells, more preferably promotion of UPK gene expression in bladder epithelial cells. The method according to <9>.
<11> A non-therapeutic UPK expression promoting method comprising administering or ingesting at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof.
<12> The method according to <11>, wherein the UPK is at least one selected from the group consisting of UPK1A, UPK1B, UPK2, and UPK3A.
<13> The daily intake of the at least one compound is 0.01 mg or more, preferably 1 mg or more, more preferably 10 mg or more, still more preferably 100 mg or more, 80000 mg or less, preferably 40000 mg or less in terms of solid matter. More preferably, it is 15000 mg or less, More preferably, it is 5000 mg or less, The method of any one of said <9>-<12>.

<14> At least one selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof as a preventive or ameliorating agent for vesicoureteral reflux or a UPK expression promoter Use of the compound.
<15> selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof for the production of a preventive or ameliorating agent for vesicoureteral reflux or a UPK expression promoter Use of at least one compound.
<16> Rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and at least one compound selected from the group consisting of coumaric acid or a salt thereof, preventing or improving vesicoureteral reflux, or promoting UPK expression Method to use as an agent.
<17> A method for preventing or ameliorating vesicoureteral reflux, or at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof, or UPK Expression promoting method.
<18> The use or method according to any one of <14> to <17>, wherein the UPK is at least one selected from the group consisting of UPK1A, UPK1B, UPK2, and UPK3A.
<19> The use according to any one of <14> and <16> to <18>, wherein the at least one compound is applied to a subject who desires prevention, amelioration, or treatment of vesicoureteral reflux disease Or method.
<20> The use or method according to any one of <14> and <16> to <19>, wherein the at least one compound is administered or ingested.
<21> Any one of the above <14> and <16> to <20>, wherein the at least one compound is divided into once to several times a day, or can be taken or administered at an arbitrary period and interval. Use or method as described.
<22> The daily intake of the at least one compound is 0.01 mg or more, preferably 1 mg or more, more preferably 10 mg or more, still more preferably 100 mg or more and 80000 mg or less, preferably 40000 mg in terms of solid matter. The use or method according to any one of <14> and <16> to <21>, more preferably 15000 mg or less, and still more preferably 5000 mg or less.
<23> The content or content of the at least one compound relative to the total amount of the agent is 0.01% by mass or more, preferably 0.1% by mass or more, more preferably 1% by mass or more, in terms of solid matter. The use or method according to any one of <14> to <16> and <18> to <22>, which is 100% by mass or less, preferably 90% by mass or less, and more preferably 70% by mass or less.
<24> The blending amount or content of the at least one compound with respect to the total amount of the agent is 0.001% by mass or more, preferably 0.01% by mass or more, more preferably 0.1% by mass in terms of solid matter. The use or method according to any one of <14> to <16> and <18> to <22>, which is 50% by mass or less, preferably 10% by mass or less, and more preferably 3% by mass or less. .
<25> The use or method according to any one of <14> and <16> to <24>, wherein the at least one compound is applied to bladder epithelial cells.
<26> The use or method according to any one of <14> to <25>, wherein the at least one compound is used non-therapeutically.

<27> At least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof, used for a method for treating vesicoureteral reflux disease.
<28> At least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof for a non-therapeutic method for treating vesicoureteral reflux use.
<29> selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof for the manufacture of a preventive or therapeutic agent for vesicoureteral reflux or a UPK expression promoter. Use of at least one compound.
<30> The use according to <29>, wherein the UPK is at least one selected from the group consisting of UPK1A, UPK1B, UPK2 and UPK3A.
<31> The compound or use according to any one of <27> to <30>, wherein the at least one compound is applied in the form of a pharmaceutical composition.
<32> The compound or use according to any one of <27> to <30>, wherein the at least one compound is applied in the form of food or beverage.
<33> The compound or use according to <32> above, wherein the form of the food or beverage is a food for a sick person, a food with a nutritional function, or a food for specified health use.
<34> The compound or use according to any one of <27> to <33>, wherein prevention or improvement of vesicoureteral reflux disease is due to UPK expression promotion.
<35> The compound or use according to any one of the above <27> to <34>, which is based on promotion of UPK expression in bladder epithelial cells, preferably promotion of UPK gene expression in bladder epithelial cells.
The blending amount or content of the at least one compound relative to the total amount of <36> agent is 0.01% by mass or more, preferably 0.1% by mass or more, more preferably 1% by mass or more, 100 in terms of solid matter. The compound or use according to any one of the above <27> to <35>, which is not more than mass%, preferably not more than 90 mass%, more preferably not more than 70 mass%.
The blending amount or content of the at least one compound with respect to the total amount of <37> agent is 0.001% by mass or more, preferably 0.01% by mass or more, more preferably 0.1% by mass or more in terms of solid matter. 50% by mass or less, preferably 10% by mass or less, more preferably 3% by mass or less, or the compound or use according to any one of <27> to <35>.
<38> The daily intake of the at least one compound is 0.01 mg or more, preferably 1 mg or more, more preferably 10 mg or more, still more preferably 100 mg or more and 80000 mg or less, preferably in terms of solid matter. The compound or use according to any one of <27> to <37>, which is administered or ingested as 40000 mg or less, more preferably 15000 mg or less, and still more preferably 5000 mg or less.

  Hereinafter, although the present invention is explained in detail based on an example, the present invention is not limited to this.

(Example 1) Examination of UPK gene expression change using human bladder epithelial cell line <Used cells>
The human bladder epithelial cell line HT-1376 (ATCC CRL-1472) was used. Detailed information of HT-1376 is shown in Table 1.

<Medium used>
MEM Earle's (manufactured by Invitrogen) to which 10% FCS (fetal calf serum), sodium pyruvate (0.055 g / 500 mL) and L-glutamine (0.146 g / 500 mL) were added was used.

In 12well plates were seeded with HT-1376 so that ^{5.0 × 10 4 cells / well,} 37 ℃ above medium, under 5% CO _2, and cultured for 48 hours. When 70% confluent, ferulic acid, coumaric acid or dimethyl sulfoxide (manufactured by Wako Pure Chemical Industries, Ltd.) was added to the medium as a control solvent, followed by further culturing for 96 hours. After the culture, RNA was extracted using RNeasy mini kit (manufactured by Qiagen).
From 100 μg of the extracted RNA, cDNA was synthesized using High Capacity RNA to cDNA kit (manufactured by Applied Biosystems) and subjected to Real-time PCR to quantify the expression levels of UPK3A gene, UPK1B gene, and UPK2 gene. UPK3A (Hs00199590_m1), UPK1B (Hs0104175_m1), and UPK2 (Hs03988771_g1) were used as Taqman Gene Expression Assay (Applied Biosystems) probes that specifically detect the target gene. The expression level of the UPK3A, UPK1B, and UPK2 genes was calculated as a relative value with respect to the expression level of each gene when the control solvent was added, assuming that the expression level of each gene when the control solvent was added was 1. The results are shown in Tables 2-4. In addition, the significant difference test was performed by the Dunnet method.

As shown in Table 2, the expression level of UPK3A gene tended to increase with the addition of ferulic acid or coumaric acid. In particular, the addition of 1000 μM ferulic acid or coumaric acid significantly increased the expression level of the UPK3A gene compared to the control.
Moreover, as shown in Table 3, the expression level of the UPK1B gene tended to increase with the addition of ferulic acid or coumaric acid. In particular, the addition of 1000 μM ferulic acid significantly increased the expression level of the UPK1B gene compared to the control.
Furthermore, as shown in Table 4, the expression level of UPK2 gene tended to increase with the addition of ferulic acid or coumaric acid. In particular, the addition of 1000 μM ferulic acid significantly increased the expression level of the UPK2 gene compared to the control.

As described above, UPK expression is significantly promoted by applying any one compound of ferulic acid and coumaric acid. This indicates that ferulic acid and coumaric acid are effective in promoting UPK expression. Furthermore, these compounds having an action of promoting the expression of UPK have been shown to be effective in preventing or ameliorating vesicoureteral reflux disease.
Rosmarinic acid has been reported to be metabolized in vivo to ferulic acid and coumaric acid (Eur. J Nutr., 2005, vol44, p.1-9, Life Science, 2004, vol75, p.165-178). Therefore, ferulic acid, coumaric acid that promotes UPK expression, and rosmarinic acid that is metabolized in vivo to ferulic acid and coumaric acid, UPK expression promoting agent, preventive or ameliorating vesicoureteral reflux disease, UPK expression promoting It can be set as the active ingredient of the food / beverage composition and the food / beverage composition for prevention or improvement of vesicoureteral reflux.

(Example 2)
SHR (42 weeks old, female, obtained from Japan SLC) was used as a rat used in the study on the effect of oral administration of rosmarinic acid on rat bladder UPK expression.
The control group includes a control diet (5% corn oil, 20% casein, 66.5% potato starch, 4% cellulose, 1% vitamin (trade name: vitamin mixture AIN-76, obtained from Oriental Bioservice)), A 3.5% mineral (trade name: mineral mixed AIN-76, obtained from Oriental Bioservice) was orally ingested for 8 weeks. Treatment group includes rosmarinic acid-containing food (5% corn oil, 20% casein, 66% potato starch, 4% cellulose, 1% vitamin, 3.5% mineral, 0.5% rosmarinic acid (Carbosyth)) Ingested for 8 weeks.
Eight weeks after ingestion, the animals were euthanized under isoflurane anesthesia and the bladder was removed.

RNA was extracted from the excised bladder using RNeasy mini kit (manufactured by Qiagen). The bladder was crushed using Tissue Lyser II (QIAGEN) in an RNA extraction solution. From 100 μg of the extracted RNA, cDNA was synthesized using High Capacity RNA to cDNA kit (Applied Biosystems) and subjected to Real-time PCR to quantify the expression levels of UPK1A gene, UPK1B gene, UPK2 gene and UPK3A gene.
Taqman Gene Expression Assay (Applied Biosystems) probes that specifically detect the gene of interest were UPK1A (Rn01480068_m1), UPK1B (Rn01447456_m1), UPK2 (Rn01418556_g1), and UPK3A82 (R101418556_g1), UPK3A (R01014856_g1), UPK3A (R101418556_82) The expression levels of the UPK1A gene, UPK1B gene, UPK2 gene, and UPK3A gene were set to 1 for the expression level of each gene in the control group, and the treatment group calculated a relative value with respect to the expression level of each gene. The results are shown in Table 5. The significant difference test was performed by Student's T test.

  As shown in Table 5, in the treatment group ingesting the rosmarinic acid-containing food, the expression levels of UPK1A gene, UPK1B gene, UPK2 gene, and UPK3A gene were significantly increased as compared with the control group ingesting the control food. did.

As described above, the oral intake of rosmarinic acid significantly promotes the expression of UPK in the bladder. This indicates that rosmarinic acid and its metabolites ferulic acid and coumaric acid are effective in promoting the expression of UPK. Furthermore, these compounds having an action of promoting the expression of UPK have been shown to be effective in preventing or ameliorating vesicoureteral reflux disease.
Therefore, rosmarinic acid, ferulic acid, and coumaric acid that promote UPK expression are UPK expression promoting agents, preventive or ameliorating agents for bladder ureteral reflux, food / beverage product compositions for promoting UPK expression, and prevention of vesicoureteral reflux Or it can be set as the active ingredient of the food-drinks composition for improvement.

Claims (6)

  An agent for preventing or improving vesicoureteral reflux, comprising as an active ingredient at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof.   Food / beverage composition for prevention or amelioration of vesicoureteral reflux disease comprising, as an active ingredient, at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof object.   A uroplakin expression promoter comprising as an active ingredient at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof.   A food and drink composition for promoting uroplakin expression comprising as an active ingredient at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof.   Non-therapeutic method for preventing or ameliorating vesicoureteral reflux disease by administering or ingesting at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof . A non-therapeutic method for promoting uroplakin expression, comprising administering or ingesting at least one compound selected from the group consisting of rosmarinic acid or a salt thereof, ferulic acid or a salt thereof, and coumaric acid or a salt thereof.