JP2007204366A - Therapeutic agent for urinary disturbance - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a therapeutic agent for two phenomena of pollakisuria and difficulty of urination. <P>SOLUTION: The therapeutic agent for urinary disturbance comprises a compound which suppresses the GIRK (G-protein conjugate type inner rectifying K ion) channel activated current while showing no suppressive effect on the glycine receptor activated current or NMDA (N-methyl-D-aspartic acid) receptor activated current. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a therapeutic agent for dysuria. The present invention relates to a therapeutic agent for dysuria for the treatment of dysuria caused by cerebrovascular disorders.

    WHO cites dysuria as well as Alzheimer's disease and osteoporosis as three major diseases to be overcome in the 21st century. This is because these three diseases for which there are still no excellent therapeutic agents are expected to continue to increase, particularly in developed countries.

  On the other hand, cerebrovascular disorders, one of the causes of dysuria, are not only stroke types that suddenly cause hemiplegia, speech impairment, and disturbance of consciousness, but also slowly, such as gait, swallowing, and intelligence. There are also types, asymptomatic types. With recent advances in diagnostic technology, it has been revealed that the incidence of slow and asymptomatic types is higher than previously thought. Urinary disturbances can occur in any of these three types, and dysuria may be the only symptom. In addition, cerebral infarction, one of the cerebrovascular disorders, has been increasing due to aging and changes in lifestyle habits.

  As a sequelae of cerebral infarction, urination disorder called neurogenic bladder occurs as frequently as unilateral hemiplegia, which is a typical sequelae.

  For urination disorders due to cerebrovascular disorders, (1) urinary urgency, urge urinary incontinence and frequent urinary incontinence that can not be tolerated if urination occurs (hereinafter referred to as “overactive bladder”), and (2) when urinating There are two dysuria functions, dysuria that cannot be urinated (hereinafter referred to as “difficulty urinating”). It is thought that (1) is due to bladder spasm, and (2) is due to bladder paralysis or increased urethral resistance. These disorders often occur in the same patient and cause great mental damage to the patient.

  Overactive bladder has been attracting attention in recent years, such as WHO promoting an international conference. In Japan, there are an estimated 8.1 million patients with overactive bladder. Several drugs have been developed for this overactive bladder.

As therapeutic agents for overactive bladder, for example, smooth muscle relaxants and anticholinergics are used. Anticholinergic drugs prevent bladder contraction and are effective for incontinence, but have the side effects of increased intraocular pressure and dry throat.
Akaike, N. & Harata, N. (1994) .Nystatin perforated patch recording and its applications to analyze of intracellular mechanisms.Jpn.J.Physiol., 44, 433-473. N. Akaike, Tetsuya Shirasaki. (1992). Nerve cell experiment method. (Ed.) Okabe Susumu. Biopharmaceutical experiment course 14. Organ function measurement method I. Yodogawa Shoten, p3-29 Koizumi, J., Yoshida, Y., Nakazawa, T. & Onodera G. (1986). Experimental studies of ischemic rats in which recirculation can be introduced in the ischemic area. Jpn. J. Stroke, 8, 1-8. Yaksh, TL, Durant, PA & Brent, CR (1986) .Micturition in rats: a chronic model for study of bladder function and effect of anesthetics. Am. J. Physiol. 251, R1177-1185.

  On the other hand, cholinergic drugs and the like are also used for difficulty in urinating, but there are many cases where difficulty in urination and overactive bladder often occur in the same patient, and no effective therapeutic agent is currently known. For this reason, development of an effective therapeutic agent for dysuria is desired. Therefore, the present invention is to provide an effective therapeutic agent for urinary dysuria, particularly dysuria due to cerebrovascular disorder.

  That is, the present invention suppresses GIRK channel activation current and urinates containing a compound that does not have an inhibitory action on glycine receptor activation current and N-methyl-D-aspartate (NMDA) receptor activation current. It is a remedy for disabilities.

  The compound is preferably at least one compound selected from the group consisting of cloperastine or its hydrochloride, or phendizoate, calamiphen hydrochloride, caramifen ethanedisulfonate, eprazinone hydrochloride, tipepidine hibenzate, and isoaminyl citrate. .

  The therapeutic agent for dysuria is preferably a therapeutic agent for dysuria caused by cerebrovascular disorder.

  From another aspect, the present invention suppresses three currents: G protein-coupled inward rectifier potassium ion channel (GIRK channel) activation current, glycine receptor activation current, and NMDA receptor activation current. This is a screening method for a dysuria drug by evaluating the presence or absence of an action to be performed.

  The therapeutic agent for dysuria represented by cloperastine in the present invention has an effect of improving urethral resistance, decreased urinary flow rate and decreased bladder capacity, and overactive bladder which is dysuria after cerebrovascular disorder It is effective in improving two symptoms of difficulty in urinating.

Conventionally, a therapeutic agent having a high effect on dysuria based on cerebrovascular disorder has not been known. The therapeutic agent for dysuria represented by cloperastine in the present invention improves an increase in urethral resistance, a decrease in urinary flow rate, and a decrease in bladder capacity. Therefore, as a therapeutic agent for neurogenic bladder caused by cerebrovascular disorder in particular. Promising.

  The therapeutic agent for dysuria of the present invention (1) suppresses GIRK channel activation current, (2) does not have an effect of suppressing glycine receptor activation current, and (3) suppresses NMDA receptor activation current It is a therapeutic agent for dysuria, comprising a compound defined by three characteristics that it has no action, particularly a compound marketed as a non-narcotic central antitussive.

The GIRK channel activation current is a membrane current resulting from the flow of potassium ions across the cell membrane due to the activation of the GIRK channel. This channel is conjugated to various neurotransmitter receptors such as serotonin (5-HT) and noradrenaline, and can be activated by, for example, serotonin and noradrenaline which stimulate 5-HT _1A receptor and adrenaline α ₂ receptor, respectively.

  In the present invention, the glycine receptor activation current is a membrane current resulting from the flow of chloride ions across the cell membrane due to activation of the glycine receptor. Thus, the glycine receptor activation current can be induced by glycine.

In the present invention, the aspartate current is an inward membrane current that flows when aspartate activates the NMDA receptor, which is one of excitatory amino acid receptors.

  In the primary screening of the therapeutic agent for dysuria in the present invention, GIRK channel activation current, glycine receptor activation current, and NMDA receptor activation current are preferably measured by the patch clamp method described in Non-Patent Document 1. This is done by measuring. There are two types of patch clamp methods: nystatin perforated patch clamp method and whole cell patch clamp method.

  An example of an apparatus used in the patch clamp method is shown in FIG. FIG. 1 is a schematic diagram showing an example of a system for measuring a membrane current in the present invention. In FIG. 1, 1 is a neuron, 2 is a glass electrode for patch clamping, 3 is a Y tube through which a test solution flows, 4 is a test tube outlet of the Y tube, 5 is a suction pump for exchanging the test solution in the tube, 6 is a culture dish, 7 is a reflux liquid inlet, 8 is a reflux liquid outlet, 9 is an amplifier, and 10 is a ground wire.

  FIG. 2 is an enlarged view of the main part of FIG. 1, 11 is a Y-tube, 12 is an ejection port, 13 is a neuron, and 14 is a recording electrode for measuring membrane current

  A method for isolating a neuron used for measuring a membrane current, particularly a single neuron of a preferred raphe nucleus and central gray matter is known per se. For example, the method proposed by Akaike et al. In Non-Patent Document 2 is adopted. be able to. In this method, for example, a thin slice of brain can be sliced from a rat brain stem and isolated by enzymatic and mechanical treatment.

  As a method for measuring the membrane current by the patch clamp method using the above apparatus, a hole patch clamp method and a nystatin perforated patch clamp method are known. In the hole patch clamp method, the recording electrode is brought into contact with the acutely isolated raphe nucleus single neuron described above to form a gigaohm seal by applying a negative pressure inside the electrode, and then applying a negative pressure to the cell membrane at the tip of the electrode. The membrane current by extracellular administration of serotonin or the like can be recorded by breaking and penetrating the electrode and the cell. In the nystatin perforated patch clamp method, as described in Non-Patent Document 1, when a recording electrode is filled with nystatin to form a gigaohm seal, the nystatin in the electrode forms a perforation in the cell membrane, and the intracellular environment is controlled. The membrane current can be recorded while maintaining it.

  A batch clamp amplifier is preferably used for recording the membrane potential and membrane current. The obtained current waveform is observed with an oscilloscope or the like, analyzed with a personal computer, and recorded on a recording medium such as a flexible disk, CD-R / RW, DVD, or MO if necessary. If necessary, the membrane current and membrane potential are recorded on a magnetic tape and drawn by a recorder. It is preferable to use a patch clamp analysis system for the analysis.

  Judgment whether or not to suppress the activation current of the GIRK channel or the like is expressed, for example, by expressing the current amplitude as an average value ± standard error and statistically comparing between the presence and absence of the drug. When the risk factor p <0.05, the difference is considered significant.

  Examples of compounds that suppress the GIRK channel activation current and do not suppress the glycine receptor activation current and the NMDA receptor activation current include, There are fans, oxerazine, pentoxyberine, eprazinone, benproperin, guaifenesin, their hydrochlorides, phosphates, citrates or phendizoates. These may be used as a mixture.

  Among these compounds, cloperastine or its hydrochloride, or phendizoate, calamiphen hydrochloride, caramifen ethanedisulfonate, eprazinone hydrochloride, tipepidine hibenzate, and isoaminyl citrate are preferred, and cloperastine is most preferred.

  Cloperastine that can be exemplified as a particularly preferred compound in the present invention is a compound represented by the following structural formula (1) (1- [2-[(4-chlorophenyl) phenylmethoxy] ethyl] piperidine), having a boiling point of 424 ° C. and a molecular weight. 330 is known as a central antitussive with mild antihistamine action.

  Caramifen, which can be exemplified as a preferred compound in the present invention, is a compound represented by the following structural formula (2), and is also known as an antiparkinsonian drug.

  Tipepidine, which can be exemplified as a preferred compound in the present invention, is a compound represented by the following structural formula (3) and is also known as an analgesic.

  All of the compounds exemplified above, including cloperastine, are commercially available as non-narcotic central antitussives and can be easily purchased from antitussives manufacturing or sales companies.

The therapeutic agent for dysuria of the present invention is administered orally (including sublingual administration) or parenterally. Examples of such drug forms include tablets, capsules, fine granules, pills, troches, infusions, injections, suppositories, ointments, patches and the like.

  As an infusion for administration of a compound into a living body, physiological saline as a main component, and other water-soluble additives and chemicals blended as necessary can be used. Additives added to such water include alkali metal ions such as potassium and magnesium, lactic acid, various amino acids, fats, glucose, fragrance, sugars and other nutrients such as vitamins A, B, C, Examples include vitamins such as D, phosphate ions, chloride ions, hormones, plasma proteins such as albumin, polymer polysaccharides such as dextrin and hydroxyethyl starch, and the like.

The concentration of the compound in such an aqueous solution is preferably in the range of 10 ⁻⁷ M to 10 ⁻⁵ M.

  The therapeutic agent containing the compound of the present invention can also be administered to a living body as a solid agent. Examples of solid agents include powders, fine granules, granules, microcapsules, tablets and the like. Among such solid preparations, it is preferable that the tablet is preferably easy to swallow.

  Known fillers such as oligosaccharides can be used as fillers and binders for forming tablets together with the compound. The tablet preferably has a diameter of 2 to 10 mm and a thickness of 1 to 5 mm. The therapeutic agent containing the compound may be mixed with other therapeutic agents.

  Various additives usually used can be blended in the solid agent. Examples of such additives include stabilizers, surfactants, solubilizers, plasticizers, sweeteners, antioxidants, flavoring agents, coloring agents, preservatives, inorganic fillers, and the like. .

  As the surfactant, anionic surfactants such as higher fatty acid soap, alkyl sulfate ester salt, polyoxyethylene alkyl ether sulfate salt, acyl N-methyl taurate salt, alkyl ether phosphate ester salt, N-acyl amino acid salt; Cationic surfactants such as alkyltrimethylammonium chloride, dialkyldimethylammonium chloride, benzalkonium chloride, alkyldimethylaminoacetic acid betaine, alkylamidodimethylaminoacetic acid betaine, 2-alkyl-N-carboxy-N-hydroxyimidazolinium betaine Nonionic surfactants such as amphoteric surfactants, polyoxyethylene types, polyhydric alcohol ester types, and ethylene oxide / propylene oxide block copolymers are available, but are not limited thereto.

  As an inorganic filler mix | blended for the objectives, such as improvement of swallowing property, talc, mica, titanium dioxide etc. can be mentioned, for example.

  Examples of the stabilizer include adipic acid and ascorbic acid. Examples of the solubilizer include surfactants such as sucrose fatty acid ester and stearyl alcohol, asparagine, arginine and the like. Examples of the sweetener include aspartame, amateur, licorice, fennel and the like.

Examples of the suspending agent include carboxyvinyl polymer, the antioxidant includes ascorbic acid, the flavoring agent includes sugar flavor, and the pH adjusting agent includes sodium citrate.

  The therapeutic agent for dysuria of the present invention is usually administered into the body in a range of 1 to 40 mg, preferably 10 to 20 mg, 3 times a day.

  The mechanism by which the therapeutic agent for dysuria of the present invention, particularly cloperastin (hereinafter sometimes collectively referred to as “cloperastin”) suppresses dysuria together with overactive bladder is not clear, but dextromelt is the same antitussive agent. There are interesting differences when compared to fans. Dextromethorphan suppresses overactive bladder but does not suppress or rather exacerbates difficulty urinating. Cloperastine, on the other hand, suppresses dysuria together with overactive bladder. Both dextromethorphan and cloperastine strongly inhibit GIRK channel activation current, whereas dextromethorphan also inhibits NDMA receptor activation current and glycine receptor activation current. On the other hand, cloperastine does not suppress NDMA receptor activation current and glycine receptor activation current.

  With respect to overactive bladder, cloperastine has an improvement effect on overactive bladder when 24 hours or more have passed after cerebral infarction. Also, dextromethorphan worsens urethral resistance and urinary flow rate with increasing dose, whereas cloperastine restores to control levels.

  When dextromethorphan and cloperastine are compared with respect to residual urine, residual urine is observed after dextromethorphan administration, but no residual urine is observed after cloperastine administration.

  EXAMPLES Next, although an Example is given and this invention is demonstrated in more detail, this invention is not restrict | limited at all to these Examples, unless the summary is exceeded.

Example 1
Changes in bladder function after cerebral infarction were analyzed using cerebral infarction model rats with middle cerebral artery (MCA) occlusion. In the analysis, a single cystometry method was adopted.

  The model rat used in the test was a Sprague-Dawley male rat weighing 250-280 g purchased from Kudo Co., Ltd. and bred in a laboratory animal facility at Kumamoto University under a room temperature of 22 ± 2 ° C. During the breeding, solid feed (CE-2) and drinking water (tap water) were freely ingested.

The reagents used in the experiments were purchased from the following.
Dextromelt fan, Cloperastine: Glycine manufactured by SIGMA, L-Aspartaic acid: Pentobarbital manufactured by Wako Pure Chemical Industries, Ltd .: Dainippon Pharmaceutical halothane: Takeda

  Anesthetized with halothane when measuring intravesical pressure on the day of infarct formation, and with pentobabital (45 mg / kg) when measuring one day after infarction. Following anesthesia, a cerebral infarction with a left middle cerebral artery embolism was prepared in rats according to the method of Koizumi.

  In the experiment, the rats were fixed in a Ballman cage as shown in FIG. 3A, and an intravesical pressure curve (CMG) was recorded without anesthesia. CMG recording was performed according to the method of Yaksh et al. As shown in FIG. 3B, the exposed end of the bladder fistula was connected to a 5 cc syringe 23 via a three-way cock 21 and a polyethylene tube 22 (PE-50, Becton Dickinson). The syringe barrel 23 was set in an infusion pump 24 (Model A, Nihon Kohden), and physiological saline kept at 37 ° C. was injected into the bladder at a rate of 0.126 ml / min. The other end of the three-way cock 21 was connected to a pressure transducer 26 (DX-360, Nihon Kohden) via a polyethylene tube 25 (PE-50, Becton Dickinson) for CMG measurement. The electric signal of the pressure transducer 26 was amplified by a biological amplifier 27 (AP-641G, Nihon Kohden) and drawn as CMG by a pen recorder 28 (RM-6100, Nihon Kohden). In order to prevent the influence of physiological saline remaining in the bladder due to an increase in urethral resistance, the physiological saline in the bladder was removed every time CMG was recorded.

  Cloperastine was administered to cerebral infarction model rats by dissolving cloperastine in physiological saline and injecting it intravenously. A single administration volume was 0.1 to 0.12 ml.

As evaluation items regarding dysuria, bladder capacity, maximum bladder pressure, urination latency, urination threshold, bladder compliance, urinary flow rate, urethral resistance, and residual urine volume were examined. Each measured value was calculated based on FIG.

The measurement results 24 hours after the infarction are shown in FIGS. FIG. 5 shows the measurement result of the maximum intravesical pressure, FIG. 6 shows the urination threshold pressure, FIG. 7 shows the urinary flow rate, FIG. 8 shows the urethral resistance, FIG. 9 shows the bladder compliance indicating the ease of extension of the bladder, FIG. 11 is a graph showing the measurement result of the urination latency. Cloperastine 5 mg / kg significantly increased bladder capacity, micturition latency and urinary flow rate, and significantly decreased urethral resistance.

(Comparative Example 1)
An infarction was formed in the same manner as in Example 1, and the evaluation items related to urination disorder were measured for model rats 24 hours after the infarction formation without administration of cloperastine. The measurement results are shown in FIGS.

(Comparative Example 2)
In Example 1, it carried out like Example 1 except administering the physiological saline which does not add cloperastine. The results are shown in FIGS.

(Example 2)
The model mice were examined for bladder capacity, maximum intravesical pressure, urination latency, urination threshold pressure, bladder compliance, urinary flow rate, and urethral resistance after 1.5-3 hours of infarction. The results are shown in FIGS. 12 shows the measurement result of the maximum intravesical pressure, FIG. 13 shows the urination threshold pressure, FIG. 14 shows the urinary flow rate, FIG. 15 shows the urethral resistance, FIG. 16 shows the bladder compliance showing the ease of extension of the bladder, FIG. 18 is a graph showing the measurement result of the urination latency. The effect of cloperastine was not as pronounced as after 24 hours of cerebral infarction.

(Comparative Example 3)
An infarct was formed in the same manner as in Example 2, and the evaluation items relating to dysuria were measured on model rats 1.5 to 3 hours after the infarction without administration of cloperastine. The measurement results are shown in FIGS.

(Comparative Example 4)
In Example 2, it carried out like Example 2 except administering the physiological saline which does not add cloperastine. The results are shown in FIGS.

(Example 3)
In measurement of GIRK channel activation current, glycine receptor activation current, and NMDA receptor activation current, acute isolation of rat raphe nucleus and central gray single neuron was first performed. Isolation of single central neurons was based on the method of Akaike et al. The method is abbreviated below. The experimental animals used were Wistar rats 10-16 days old (purchased from Kudo Co., Ltd.) without discrimination between males and females. The rats were lightly anesthetized with Nembutal injection, decapitated, and the brainstem was quickly removed. Next, a brain slice was prepared with a thickness of 400 μm using a micro slicer (DTK-1000, Dosaka EM).

  The obtained brain slices were left for 30-60 minutes at room temperature in Krebs solution in which 95% oxygen and 5% carbon dioxide were bubbled. Thereafter, the enzyme treatment was carried out at 31 ° C. for 30-40 minutes in a Pronace (1 mg / 9 ml) solution and then at 31 ° C. for 20 minutes in a thermolysin (1 mg / lOml) solution. After leaving the enzyme in the Krebs solution for about 60 minutes, use the microfeather scalpel or No. 11 scalpel under the stereomicroscope (MS-5, Leica) and the raphe nucleus region or caudal ventrolateral central gray matter Was cut out.

  The sliced brain thin section was transferred to a 35 mm culture dish (Primaria3801, Becton Dickinson) filled with Krebs solution, and the brain diameter was observed with an inverted microscope (DM-IL, Leica). Dispersed mechanically using a 500 μm glass pipette, raphe nuclei and caudal ventrolateral central gray matter single neurons as shown in FIG. 19 were obtained. All these operations were performed at room temperature.

  The apparatus shown in FIG. 1 was used to measure the GIRK channel activation current, glycine receptor activation current, and NMDA receptor activation current. The drug was dissolved in HEPES buffered extracellular solution and administered by the Y-tube method. One end of the Y-tube was led to a chemical reservoir, and the other end was connected to a suction pump via a solenoid valve and a decompression bottle. Administration of the chemical solution is performed by opening and closing the solenoid valve, and when the solenoid valve opens, the chemical solution is drawn into the Y-tube, and the chemical solution is ejected from the tip due to the water pressure difference between the chemical solution level and the tip of the Y-tube. .

  The tip of the Y-tube is approximately 200 μm in the horizontal direction from the center of the cell body of the cells to be recorded using a three-dimensional micromanipulator and about 50 μm in height from the bottom of the culture dish. The nerve cells were placed at the center of the flow (FIG. 2).

  Membrane current was measured in a patch clamp whole cell recording format. After confirming that the neurons isolated under the inverted microscope (TMD, Nikon) were attached to the bottom of the culture dish, applying a rectangular wave voltage pulse to the recording electrode, the three-dimensional hydraulic micromanipulator (MHW-3, Narishige), the electrode tip was brought into light contact with the cell surface, and a gigaohm seal was formed by suction. When measuring with the nystatin perforated patch clamp method, after waiting for about 1-10 minutes after forming the gigaohm seal, nystatin forms a small ion-permeable pore in the cell membrane directly under the recording electrode, and a whole-cell recording format is obtained. It was. In the case of measuring by the conventional whole cell clamp method, after forming the gigaohm seal, the inside of the recording electrode was further subjected to a negative pressure, thereby destroying the cell membrane immediately below the recording electrode and obtaining the whole cell recording format.

  A batch clamp amplifier (Axopatch 1D, Axon Instruments) was used for recording the membrane current. The obtained current waveform was observed with an oscilloscope (V-225, Hitachi) and recorded on a personal computer (DOS / V specification) online. At the same time, it was also stored on a video tape (HV-FR30, Aiwa) via a PCM processor (PCM Data Recording System Model RP-880, NF Electronic Institute). The records stored on the computer were drawn with a laser printer, and the records stored on the videotape were drawn with a thermal array coder (RTA-1200, Nibon Kohdcn).

  The recording glass electrode (recording electrode) was produced from a glass tube having an outer diameter of 1.5 mm and an inner diameter of 0.9 mm using an electrode production device (PP-83, Narishige). A tip having an inner diameter of about 1 μm and a DC resistance of about 6 MΩ when the electrode was filled with internal liquid was used.

For recording by the nystatin perforated patch clamp method, an in-electrode solution having the following composition was used. 75 mM KCl, 60 mM K-gluconate, 10 mM HEPES, nystatin 100-200 μg / ml, pH 7.2. For recording by the whole cell patch clamp method, an in-electrode solution having the following composition was used. 70 mM KCl, 70 mM K-gluconate, 4 mM EGTA, 5 mM MgCl ₂ , 4 mM Na ₂ -ATP, 0.3 mM GTPγS, 10 mM HEPES, pH 7.0.

The nystatin stock solution was prepared as follows. First, 10 mg of nystatin per ml of methanol was added and well dispersed using an ultrasonic cleaner. Then, 1N HCl was added while stirring the solution with a stirrer to dissolve nystatin well. Finally, 1N NaOH was added dropwise to adjust the pH to about 7.0.

(Reference Example 1)
Glycine receptor activation current measurement test by the method described in Example 3 using neurons isolated acutely from the ventrolateral part of the PAG region of one of the urination centers as neurons and using 10 ⁻⁴ M glycine as a drug solution Went. 10 ⁻⁴ M glycine caused a rapidly activated inward current at a holding potential of −60 mV. (Fig. 20)

Example 4
In Reference Example 1, the procedure was the same as Reference Example 1 except that 3 × 10 ⁻⁴ M cloperastine was pretreated for 30 seconds in advance before administration of glycine, and glycine and cloperastine were immediately administered simultaneously. Cloperastine did not suppress this glycine receptor activation current. (FIGS. 20 and 21)

(Comparative Example 5)
The same procedure as in Example 4 was carried out except that dextromethorphan was used in place of cloperastine and 3 × 10 ⁻⁵ M was used as the glycine concentration. The results are shown in FIG. Dextromethorphan suppressed glycine activation current.

(Reference Example 2)
The dorsal raphe nucleus was used as a neuron. Using 3 × 10 ⁻⁴ M aspartic acid as a drug solution, a measurement test of aspartate-induced NMDA receptor activation current was performed by the method described in Example 3. Aspartate current was recorded in the absence of Mg ^{2+ in the} presence of 10 ⁻⁶ M glycine. Pre-treatment with 10 ⁻⁶ M glycine for 30 seconds at a holding potential of −60 mV followed by administration of aspartic acid and glycine recorded a fast-acting inward current. (Fig. 23)

(Example 5)
In Reference Example 2, the same procedure as in Reference Example 2 was performed, except that pretreatment with 10 ⁻⁶ M glycine and 10 ⁻⁵ M cloperastin was performed for 30 seconds before administration of aspartic acid. The obtained results are shown in FIG. Cloperastine did not inhibit aspartate-induced NMDA receptor activation current.

(Comparative Example 6)
One example of the result of the inhibitory action of dextromethorphan on the NMDA receptor activation current is shown in FIG. In the experiment shown in this figure, the NMDA receptor activation current was expressed by NMDA itself under the condition of a solution containing glycine 10 ⁻⁶ M. The concentration of NMDA used is 5 × 10 ⁻⁵ M.

(Reference Example 3)
(GIRK channel activation current)
The same procedure as in Example 3 was performed except that serotonin was administered in an extracellular solution containing 20 mM K ⁺ and 1.2 mM Mg ²⁺ . 10 ⁻⁹ M to 3 × 10 ⁻⁶ M serotonin generated a GIRK channel activation current at a holding potential of −80 mV. (Fig. 25)

(Example 6)
(Effect of cloperastine on serotonin-induced GIRK channel activation current)
In Reference Example 3, the same procedure as in Reference Example 3 was performed, except that 3 × 10 ⁻⁷ M to 10 ⁻⁵ M of cloperastin was pretreated for 1 minute before serotonin was administered, and serotonin and cloperastine were immediately administered simultaneously. The effect was examined. 10 ⁻⁵ M cloperastine inhibited GIRK channel activation current coupled with 5-HT _1A receptor by almost 100%. (Fig. 26)

(Comparative Example 7)
The same procedure as in Example 6 was performed except that dextromethorphan was used instead of cloperastine. The results are shown in FIG. 10 ⁻⁶ M to 3 × 10 ⁻⁵ M dextromelt fans suppressed GIRK channel currents coupled with serotonin receptors.

Dextromethorphan and Cloperastine inhibits 5-HT1A receptor activation current, the activation of the inwardly rectifying K ion channel of this effect 5-HT _1A receptor conjugated G protein-coupled to the body (GIRK channel) By suppressing the current. The operation will be described with reference to FIG. FIG. 28 is a graph showing the results of measuring the GIRK channel activation current. That is, under the condition where GTP-γ-S is circulated into cells, administration of 5-HT continuously activates the βγ subunit of G protein, and as shown in FIG. 28, the inward GIRK channel The activation current flows continuously. It can be seen that this current continues even after 5-HT is removed, and is persistent. As shown in FIG. 28, under this state, the current is not suppressed by spiperone (Spi) that blocks 5-HT _1A receptor and 5-HT ₂ receptor, but dextromelolphan (DM) and cloperastine (Clo ) Is suppressed. That is, this experimental result shows that dextromethorphan and spiperone suppressed the activation current of the GIRK channel conjugated to this receptor, not the 5-HT _1A receptor.

  The present invention relates to a therapeutic agent for dysuria. The present invention relates to a therapeutic agent for dysuria for the treatment of dysuria caused by cerebrovascular disorders.

FIG. 1 is a schematic view showing an example of an apparatus used in the patch clamp method. FIG. 2 is an enlarged view of a main part of an apparatus used in the patch clamp method of FIG. FIG. 3A is a photograph showing a method of fixing a rat in an example. FIG. 3B is a schematic diagram showing a rat cystometrogram method performed in the examples. FIG. 4 is a diagram showing measurement positions for calculating each evaluation item in the intravesical pressure curve. FIG. 5 is a graph showing the measurement result of the maximum intravesical pressure. FIG. 6 is a graph showing measurement results of the urination threshold pressure. FIG. 7 is a graph showing the measurement result of the urine flow rate. FIG. 8 is a graph showing measurement results of urethral resistance. FIG. 9 is a graph showing measurement results of bladder compliance. FIG. 10 is a graph showing measurement results of bladder capacity. FIG. 11 is a graph showing a measurement result of urination latency. FIG. 12 is a graph showing the measurement result of the maximum intravesical pressure. FIG. 13 is a graph showing measurement results of the urination threshold pressure. FIG. 14 is a graph showing the measurement result of the urine flow rate. FIG. 15 is a graph showing measurement results of urethral resistance. FIG. 16 is a graph showing measurement results of bladder compliance. FIG. 17 is a graph showing measurement results of bladder capacity. FIG. 18 is a graph showing a measurement result of urination latency. FIG. 19 is an example of a micrograph of a raphe nucleus used in the example of the present invention. FIG. 20 is a diagram showing measurement results of glycine activation current. FIG. 21 is a diagram showing measurement results of glycine activation current. FIG. 22 is a diagram showing measurement results of glycine activation current. FIG. 23 is a diagram showing a measurement result of the NMDA activation current. FIG. 24 is a diagram showing the measurement results of the NMDA activation current. FIG. 25 is a diagram showing a measurement result of the GIRK channel current. FIG. 26 is a diagram showing a measurement result of the GIRK channel current. FIG. 27 is a diagram showing a measurement result of the GIRK channel current. FIG. 28 is a diagram showing a measurement result of the GIRK channel current.

Claims (4)

  G protein-coupled inward rectifier potassium ion channel (GIRK channel) activation current is suppressed, glycine receptor activation current and N-methyl-D-aspartate (NMDA) receptor activation current are suppressed A therapeutic agent for dysuria, which contains a compound that does not have an action to act.   The above compound is at least one compound selected from the group consisting of cloperastine or its hydrochloride, or phendizoate, calamiphen hydrochloride, caramifen ethanedisulfonate, eprazinone hydrochloride, tipepidine hibenzate, and isoaminyl citrate. The therapeutic agent for dysuria according to claim 1   The therapeutic agent for dysuria according to claim 1 or 2, wherein the therapeutic agent for dysuria is a therapeutic agent for dysuria caused by cerebrovascular disorder. Urination by evaluating the presence or absence of an action that suppresses three currents: G protein-coupled inward rectifier potassium ion channel (GIRK channel) activation current, glycine receptor activation current, and NMDA receptor activation current Screening method for drugs for treating disorders.

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