JP2019214618A - Uroplakin expression promoter - Google Patents

Abstract

To provide an uroplakin expression promoter which promotes uroplakin expression in a bladder epithelial cell, and a preventive or an improver of frequent urination, urinary urgency, urinary incontinence or bladder pain which makes good use of efficacy of the uroplakin expression promoter or as its administering means, and a preventive or an improver of vesicoureteral reflux.SOLUTION: There are provided an uroplakin expression promoter having glucosamine, pyruvic acid and at least one type from the group consisting of salt thereof as an active ingredient, a preventive or an improver of frequent urination, urinary urgency, urinary incontinence or bladder pain, and a preventive or an improver of vesicoureteral reflux.SELECTED DRAWING: None

Description

  The present invention relates to a uroplakin expression promoter.

It is known that dysuria such as frequent urination, urgency, urinary incontinence, etc. greatly impairs daily life and deteriorates quality of life. In particular, in Japan, a super-aging society, it is estimated that about half of those aged 40 and over have frequent urinary symptoms.
As one of the causes of dysuria such as frequent urination, urgency, urinary incontinence, etc., it has been suggested that the epithelial barrier function of the bladder deteriorates. In patients with detrusor overactivity (indicating overactive bladder) and interstitial cystitis patients having dysuria as described above, uroplakin (hereinafter referred to as “UPK”) suggested to be associated with the epithelial barrier function of the bladder has been suggested. 3A) has been reported to be reduced (see Non-Patent Document 1). In addition, it has been reported that in UPK2 / UPK3A double knockout mice, penetration of urine into bladder tissue is enhanced (see Non-Patent Document 2). Further, it has been reported that knockout mice of UPK2 or UPK3A induce pollakiuria (see Non-Patent Document 3).

Vesicoureteral reflux is a phenomenon in which urine stored in the bladder flows back to the ureter or the renal pelvis / renal parenchyma due to the structure and abnormalities of the vesicoureteral and subsequent parts. The presence of reflux can cause recurrent pyelonephritis due to ascending urinary tract infection. Also, urine that has flowed back into the upper urinary tract during urination, descends into the bladder after urination, and becomes residual urine. Therefore, bacteria grow and cystitis develops, causing bladder pain.
One of the causes of vesicoureteral reflux has been suggested to involve UPK3A. In fact, it has been reported that vesicoureteral reflux occurs in UPK3A knockout mice (see Non-Patent Document 4).

Currently, anticholinergic drugs are used for drug treatment of overactive bladder. Anticholinergics inhibit the action of acetylcholine, which promotes contraction of the detrusor muscle of the bladder. However, anticholinergic drugs have many problems such as no clear symptom-improving effect for some patients, and symptoms such as dry mouth and constipation as side effects, and more effective prevention and improvement methods are desired. I have.
In addition, antidepressants and antihistamines have been used for drug treatment of interstitial bladder. However, since the effectiveness of antidepressants and antihistamines is limited, more effective prevention and improvement methods are desired.
In addition, for treatment of vesicoureteral reflux, conservative therapy that waits for spontaneous elimination is taken in mild cases, whereas antireflux surgery is performed when it does not disappear. However, surgical treatment is highly invasive, and a non-invasive prevention / improvement method is desired.

BJU INTERNATIONAL, 2007, vol. 39, p. 1506-1516 The Journal of Biological Chemistry, 2012, vol. 287, p. 11011-11017 Neurourology and Urodynamics, 2009, vol. 28, p. 1028-1033 Journal of Cell Biology, 2000, vol. 151, p. 961-971

Anticholinergic drugs used to treat overactive bladder inhibit the release of acetylcholine from binding to receptors in bladder smooth muscle, but this is a so-called symptomatic treatment, and a sufficient treatment from the point of effect Is hard to say. Anticholinergics also have the side effects described above. Antidepressants and antihistamines used in the treatment of interstitial cystitis inhibit neurotransmission and histamine release, but this is also a so-called symptomatic treatment, and it is hard to say that it is an effective treatment in terms of effectiveness . Furthermore, antireflux surgery performed in the treatment of cystoureteric reflux that does not spontaneously disappear is highly invasive, and there is no noninvasive and effective treatment.
In contrast, overactive bladder, interstitial cystitis, if the epithelial barrier function of the bladder considered to be a cause of vesicoureteral reflux can be suppressed, overactive bladder, interstitial cystitis, It can be a causal treatment for vesicoureteral reflux.

An object of the present invention is to provide a UPK expression promoter that promotes the expression of UPK in bladder epithelial cells.
The present invention also provides a preventive or ameliorating agent for urinary frequency, urinary urgency, urinary incontinence or bladder pain, and a preventive or ameliorating vesicoureteral reflux, which utilizes the efficacy of the UPK expression promoter and as a means of administration thereof The task is to provide agents.

Another object of the present invention is to provide a UPK expression promoting method for promoting UPK expression in bladder epithelial cells.
Another object of the present invention is to provide a non-therapeutic method for preventing or improving pollakiuria, urgency, urinary incontinence, bladder pain or vesicoureteral reflux.

In view of the above problems, the present inventors have conducted intensive studies on substances capable of suppressing a decrease in the epithelial barrier function of the bladder that causes overactive bladder, interstitial cystitis, and vesicoureteral reflux. As a result, glucosamine, pyruvic acid and their salts have the effect of promoting the expression of the UPK gene, and are useful for preventing or improving pollakiuria, urgency, urinary incontinence, bladder pain and vesicoureteral reflux Was found.
The present invention has been completed based on these findings.

The present invention relates to a UPK expression promoter comprising at least one active ingredient selected from the group consisting of glucosamine, pyruvic acid and salts thereof.
The present invention also relates to a preventive or ameliorating agent for pollakiuria, urinary urgency, urinary incontinence or bladder pain, comprising at least one active ingredient from the group consisting of glucosamine, pyruvic acid and salts thereof.
The present invention also relates to a preventive or ameliorating agent for vesicoureteral reflux, comprising at least one active ingredient selected from the group consisting of glucosamine, pyruvic acid and salts thereof.

The present invention also relates to a non-therapeutic UPK expression promoting method comprising administering or ingesting at least one member selected from the group consisting of glucosamine, pyruvic acid and salts thereof.
The present invention also relates to a method for preventing or ameliorating non-therapeutic pollakisuria, urgency, urinary incontinence or bladder pain, comprising administering or ingesting at least one member selected from the group consisting of glucosamine, pyruvic acid and salts thereof.
The present invention also relates to a non-therapeutic method for preventing or ameliorating vesicoureteral reflux, which comprises administering or ingesting at least one member selected from the group consisting of glucosamine, pyruvic acid and salts thereof.

The UPK expression promoting agent of the present invention can promote UPK expression in bladder epithelial cells.
Further, the preventive or ameliorating agent for pollakiuria, urgency, urinary incontinence or bladder pain, and the preventive or ameliorating agent for vesicoureteral reflux of the present invention include pollakiuria, urgency, urinary incontinence, bladder pain or bladder Ureteral reflux can be prevented or ameliorated.

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.

“UPK” in the present specification is a membrane protein specifically expressed in urothelium. Up to now, UPK1A, UPK1B, UPK2 and UPK3A are known as UPK1A. Among them, the present invention is particularly effective for promoting the expression of UPK2 and UPK3A.
The nucleotide sequence information and amino acid sequence information of human UPK2 and UPK3A are registered as NCBI Reference Sequence: NM_006760.3, NP_006751.1 and NCBI Reference Sequence: NM_001167574.1, NP_001161046.1, respectively, and are available from NCBI. is there. In addition, these variants or homologs can be targets for the expression promotion of the present invention.

The UPK expression promoter of the present invention, an agent for preventing or improving urinary frequency, urinary urgency, urinary incontinence or bladder pain, and an agent for preventing or ameliorating vesicoureteral reflux (hereinafter, urinary frequency, urgency, urinary incontinence or The agent for preventing or ameliorating bladder pain and the agent for preventing or ameliorating vesicoureteral reflux are also referred to as “the agent for preventing or ameliorating the present invention”) at least one of the group consisting of glucosamine, pyruvic acid and salts thereof. One type is defined as an active ingredient.
The salt of glucosamine used in the present invention can be appropriately selected. Specific examples include glucosamine hydrochloride and glucosamine sulfate. The pyruvic acid salt used in the present invention can be appropriately selected. Specific examples include sodium pyruvate, potassium pyruvate and calcium pyruvate.
For the UPK expression promoter of the present invention and the preventive or ameliorating agent of the present invention, any of glucosamine, pyruvic acid and salts thereof may be used alone, or two or more thereof may be used in combination.

The method for producing glucosamine, pyruvic acid and salts thereof is not particularly limited, and 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.
Glucosamine and its salts can be produced, for example, by hydrolyzing chitin with concentrated hydrochloric acid with reference to the method described in Fine Chemical (CMC Publishing, 2008, p. 84).
Pyruvic acid and salts thereof are produced, for example, by culturing glucose under aerobic conditions with pyruvic acid-producing bacteria belonging to the genus Microbacterium with reference to the method described in JP-A-2006-254863. can do.

As described above, it has been reported that the expression level of UPK3A is decreased in patients with detrusor overactivity or interstitial cystitis having dysuria. It has been reported that knockout mice of UPK2 or UPK3A induce pollakiuria. Furthermore, it is reported that vesicoureteral reflux develops in UPK3A knockout mice.
From these reports, it is thought that by promoting UPK expression, frequent urination, urgency, urinary incontinence and bladder pain can be prevented or ameliorated. In addition, it is considered that vesicoureteral reflux can be prevented or improved by promoting the expression of UPK.

As will be shown in Examples below, glucosamine, pyruvic acid and salts thereof have an action of promoting UPK gene expression in bladder epithelial cells. Thus, these compounds can be used to prevent or ameliorate pollakiuria, urgency, urinary incontinence and bladder pain. These compounds can also 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 UPK expression promoting agent of the present invention is one of the specific embodiments of the above-mentioned use, and can be applied to both therapeutic uses (medical uses) and non-therapeutic uses (non-medical uses). Specifically, it can be used as a drug, a quasi-drug, and the like, and the UPK expression promoter of the present invention can be blended in various foods and drinks, feeds, pet foods, and the like.
Further, the preventive or ameliorating agent of the present invention is one of the specific embodiments of the above-mentioned use, and can be applied to any of the therapeutic use (medical use) and the non-therapeutic use (non-medical use). 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.
The UPK expression promoting agent and the preventive or ameliorating agent of the present invention are applicable to humans and animals, such as liquid, solid, emulsion, paste, gel, powder (powder), granule, pellet, and stick. Various dosage forms can be taken.
Further, the UPK expression promoting agent and the preventive or ameliorating agent of the present invention may be composed of only the above-mentioned compound which is an active ingredient, or may contain other components as long as the effect is not affected. You may. Examples of the other components include the following additives.

When the UPK expression promoter of the present invention and the preventive or ameliorating agent of the present invention are applied to pharmaceuticals and quasi-drugs, the compound should be contained in an effective amount, and if necessary, additives may be added to prepare various dosage forms. Can be. For example, tablets, coated tablets, capsules, granules, powders, syrups, enteric agents, troches, drinks and other oral medicines, or injections, suppositories, transdermal absorbers, external preparations and the like It can be prepared as an oral medicament. Of these forms, the preferred form is oral medicine.
In order to prepare various dosage forms, it may be produced according to a conventional method using additives. As the additive, a commonly used additive can be used. Examples of additives include pharmaceutically acceptable excipients (sorbitol, glucose, lactose, dextrin, sugars such as starch, inorganic substances such as calcium carbonate, crystalline cellulose, distilled water, sesame oil, corn oil, olive oil, rapeseed oil, and the like). ), Liquid carriers (distilled water, physiological saline, aqueous glucose solution, alcohols such as ethanol, propylene glycol, polyethylene glycol, etc.), oily carriers (various animal and vegetable oils, white petrolatum, paraffin, waxes, etc.), stabilizers , Wetting agents, emulsifiers, binders, isotonic agents, disintegrants, lubricants, bulking agents, surfactants, dispersants, suspending agents, diluents, osmotic pressure adjusting agents, pH adjusting agents, preservatives, Examples include antioxidants, coloring agents, ultraviolet absorbers, humectants, thickeners, brighteners, buffers, preservatives, flavoring agents, fragrances, coating agents, odor correctors, and bacteria inhibitors.

When the UPK expression promoter of the present invention and the preventive or ameliorating agent of the present invention are formulated and applied to foods and drinks, feeds, pet foods and the like, forms suitable for food or drink, for example, granules, granules, tablets, capsules, It can be provided in the form of a paste or the like. Furthermore, the food or drink is a concept of preventing or improving overactive bladder, frequent urination, urgency, or urge urinary incontinence, and a beauty food, a food for the sick, and a nutritional function indicating the concept as necessary. It can be in the form of food or a functional food or drink such as food for specified health use.
Examples of blending into foods and drinks include bread, processed flour such as noodles, porridge, cooked rice and other processed rice, biscuits, cakes, jelly, chocolate, rice crackers, confectionery such as ice cream, Tofu, processed soybean foods such as processed foods, soft drinks, fruit drinks, milk drinks, beverages such as carbonated drinks, dairy products such as yogurt, cheese, butter, milk, soy sauce, sauces, miso, mayonnaise, dressing, etc. Examples include seasonings, meat storage such as ham, bacon, sausage, processed meat food, processed fishery products such as canned hampan, chikuwa and fish, cooking oil, and frying oil. In addition, tablet foods such as tablets (tablets) and capsules, thick liquid foods, natural liquid foods, semi-digestive nutritional foods, component nutritional foods, drink nutritional foods and other oral enteral nutritional foods, functional foods, etc. Good.
Examples of the feed include feed for small animals used for rabbits, rats, mice and the like, pet food used for dogs, cats, small birds, squirrels and the like.
These foods and drinks, feeds, pet foods, and the like contain the UPK expression promoter of the present invention or the preventive or ameliorating agent of the present invention, and may contain food ingredients such as sweeteners, coloring agents, antioxidants, and vitamins. It can be prepared according to a conventional method by appropriately combining additives such as foods, flavors, and minerals, proteins, lipids, carbohydrates, carbohydrates, and dietary fibers.

The amount of the active ingredient in the UPK expression promoter of the present invention and the preventive or ameliorating agent 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 and capsules, oral liquid preparations such as internal liquids and syrups, the solid content concentration (solid content conversion) is 0.001% by mass. The above is preferred, the content is more preferably 0.01% by mass or more, preferably 100% by mass or less, and more preferably 95% by mass or less. Alternatively, 0.001 to 100% by mass is preferable, and 0.01 to 95% by mass is more preferable.
When the UPK expression promoter of the present invention and the preventive or ameliorating agent of the present invention are blended in foods and drinks, pet foods, and the like, the amount of the active ingredient is preferably 0.001% by mass or more as a solid content concentration, and 0.1% by mass. The content is more preferably 01% by mass or more, preferably 50% by mass or less, and more preferably 10% by mass or less. Alternatively, 0.001 to 50% by mass is preferable, and 0.01 to 10% by mass is more preferable.

  The administration or intake of the UPK expression promoter of the present invention and the prophylactic or ameliorating agent of the present invention can be appropriately selected and determined according to the condition, weight, sex, age or other factors of the individual. For example, the daily administration or intake of an adult (60 kg) is preferably 0.001 mg or more, more preferably 1 mg or more, still more preferably 100 mg or more, and more preferably 80,000 mg or less, in terms of solid content of the compound as the active ingredient. Is preferably 40000 mg or less, and more preferably 15000 mg or less. Alternatively, 0.001 to 80000 mg is preferable, 1 to 40000 mg is more preferable, and 100 to 15000 mg is further preferable. Further, the UPK expression promoter of the present invention and the prophylactic or ameliorating agent of the present invention can be ingested / administered once to several times a day, or at any time and interval.

  Although not particularly limited as a target for ingestion or administration of the above-mentioned medicines, quasi-drugs or foods, overactive bladder, and its main symptoms, urinary frequency, urgency, urgency incontinence, etc. Desired humans and non-human mammals are preferred. In addition, the subjects to be ingested or administered are humans and non-human mammals with overactive bladder symptoms, humans and non-human mammals at risk of such symptoms, and humans and humans expecting prevention of their diseases and symptoms. Other mammals are also included.

  With respect to the above-mentioned embodiment, the present invention further discloses the following UPK expression promoter, preventive or ameliorating agent, and method.

<1> A UPK expression promoter comprising at least one active ingredient selected from the group consisting of glucosamine, pyruvic acid and salts thereof.
<2> A preventive or ameliorating agent for pollakiuria, urinary urgency, urinary incontinence or bladder pain, comprising as an active ingredient at least one member selected from the group consisting of glucosamine, pyruvic acid and salts thereof.
<3> The preventive or ameliorating agent according to <2>, wherein the prevention or amelioration of urinary frequency, urgency, urinary incontinence or bladder pain is due to improvement of overactive bladder or cystitis.
<4> An agent for preventing or ameliorating vesicoureteral reflux, comprising at least one active ingredient selected from the group consisting of glucosamine, pyruvic acid and salts thereof.

<5> A non-therapeutic UPK expression promoting method comprising administering or ingesting at least one member from the group consisting of glucosamine, pyruvic acid and salts thereof.
<6> A method for preventing or ameliorating non-therapeutic pollakisuria, urgency, urinary incontinence or bladder pain, comprising administering or ingesting at least one member from the group consisting of glucosamine, pyruvic acid and salts thereof.
<7> The method according to <6>, wherein the prevention or improvement of pollakiuria, urgency, urinary incontinence or bladder pain is due to improvement of overactive bladder or cystitis.
<8> A non-therapeutic method for preventing or ameliorating vesicoureteral reflux, which comprises administering or ingesting at least one member selected from the group consisting of glucosamine, pyruvic acid and salts thereof.
<9> For humans and non-human mammals with overactive bladder symptoms, humans and non-human mammals at risk of such symptoms, and humans and non-human mammals expecting prevention of their diseases and symptoms The method according to any one of the above <5> to <8>.

  EXAMPLES Hereinafter, although this invention is demonstrated further in detail based on an Example, this invention is not limited to this.

Examination of changes in UPK gene expression using human bladder epithelial cell line <Cells used>
HT-1376 (ATCC), a human bladder epithelial cell line, was used. Table 1 shows detailed information of HT-1376.

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

HT-1376 was seeded on a 12-well plate at 5.0 × 10 ⁴ cells / well, and cultured in the above medium at 37 ° C. under 5% CO ₂ for 48 hours. At 70% confluence, glucosamine hydrochloride, pyruvic acid or DEPC treated water (RNase-free Water, Nippon Gene) as a control was added to the medium, and the cells were further cultured for 96 hours. After the culture, RNA was extracted using an RNeasy mini kit (manufactured by Qiagen).
From 100 μg of the extracted RNA, cDNA was synthesized using a High Capacity RNA to cDNA kit (manufactured by Applied Biosystems) and subjected to Real-time PCR to quantify the expression levels of the UPK2 gene and UPK3A gene. The expression level of the UPK2 gene and UPK3A gene was calculated as a relative value to the expression level of each gene when the control was added, with the expression level of each gene when the control was added as 1. The results are shown in Table 2. The significance test was performed by the Dunnet method.

  As shown in Table 2, the addition of glucosamine hydrochloride significantly increased the expression level of the UPK2 gene as compared to the control. In addition, the addition of pyruvate significantly increased the expression level of the UPK3A gene as compared to the control.

  As described above, the expression of UPK is significantly promoted by applying any one of glucosamine, pyruvic acid and salts thereof. This indicates that glucosamine, pyruvic acid and salts thereof 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 improving pollakiuria, urgency, urinary incontinence, bladder pain, and vesicoureteral reflux overactive bladder. .

Claims (7)

  An uroplakin expression promoter comprising at least one selected from the group consisting of pyruvic acid and salts thereof as an active ingredient.   An agent for preventing or improving pollakiuria, urgency, urinary incontinence or bladder pain, comprising at least one selected from the group consisting of pyruvic acid and salts thereof as an active ingredient.   The preventive or ameliorating agent according to claim 2, wherein the prevention or improvement of frequent urination, urgency, urinary incontinence or bladder pain is due to improvement of overactive bladder or cystitis.   An agent for preventing or improving vesicoureteral reflux, comprising as an active ingredient at least one selected from the group consisting of pyruvic acid and salts thereof.   A food or drink composition for preventing or improving pollakiuria, urinary urgency, urinary incontinence or bladder pain, comprising at least one selected from the group consisting of pyruvic acid and salts thereof as an active ingredient.   The food and drink composition according to claim 5, wherein the prevention or improvement of frequent urination, urgency, urinary incontinence or bladder pain is due to improvement of overactive bladder or cystitis.   A food and drink composition for preventing or improving vesicoureteral reflux, comprising as an active ingredient at least one selected from the group consisting of pyruvic acid and salts thereof.