JPH10245336A - Inhibitor to na/ca-exchange system - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a composition inhibiting Na<+> /Ca<2+> -exchange system and useful for the treatment and prevention of ischemic heart diseases, brain diseases, and kindney diseases, and cell protection in e.g. thrombolytic therapy, by including benzyloxyphenoxyaniline as the active ingredient. SOLUTION: This inhibitor, a composition inhibiting Na<+> /Ca<2+> -exchange system, contains as the active ingredient, 2-(4-benzyloxyphenoxy)aniline shown by the formula or its salt (e.g. a salt with an inorganic acid each as hydrochloric acid, hydrobromic acid, or sulfuric acid, or a salt with sulfonic acid such as methanesulfonic acid or p-toluenesulfonic acid), and can be used orally or parenterally using, as appropriate well known support or diluent in a suitable pharmaceutical composition form of solid preparation such as tables, pills, capsules, granules, or dry syrup, or injection liquid. The solid preparation can be prepared using various additives such as vehicle, disintegrating agent, binder, lubricant, and coating base, by the agitation-granulation method, the fluidized- bed granulation method, or the crushing-granulation method.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a Na ⁺ / Ca ²⁺ exchange inhibitor. More specifically, useful treatment and prevention for ischemic heart disease such as myocardial infarction, ischemic brain disease such as cerebral infarction or ischemic renal disease, or when performing thrombolytic therapy, angioplasty, coronary artery bypass surgery, and organ transplantation Cell protection.

[0002]

2. Description of the Related Art In ischemic diseases such as myocardial infarction and cerebral infarction, blood circulation such as thrombolytic therapy or angioplasty is intended to protect cells in the ischemic area and reduce the infarct size and improve the prognosis. Therapies are widely practiced and their usefulness has been highly evaluated. However, on the other hand, so-called ischemia / reperfusion injury, in which cell injury is rather exacerbated by reperfusion, has become a problem, and a countermeasure is desired. Sufficient effects on perfusion injury have not been obtained (Japanese clinical practice, Vol. 52, pp. 1080-1087, 1994).

[0003]

SUMMARY OF THE INVENTION The object of the present invention is
⁺ / Ca ²⁺ exchange system, and thus useful treatment and prevention for ischemic heart disease such as myocardial infarction, ischemic brain disease such as cerebral infarction or ischemic renal disease, or thrombolytic therapy, angioplasty, It is intended to help protect cells during coronary artery bypass surgery and organ transplantation.

[0004]

As one of the most important mechanisms of ischemia / reperfusion injury, excessive accumulation of intracellular Ca ²⁺ via a Na ⁺ / Ca ²⁺ exchange system (so-called Ca ²⁺ overload). Lord) theory is considered promising [Journal of Molecular Cellular Cardiology, (J. Mol.
Cell. Cadiol. ) Volume 27, page 53-No. 6
Page 3, 1995. ]. During ischemia / reperfusion, Na ⁺ /
The activation of the H ⁺ exchange system and the suppression of Na ⁺ -K ⁺ ATPase increase the intracellular Na ⁺ concentration.
The a ⁺ / Ca ²⁺ exchange system is activated, and Ca ²⁺ excessively flows into cells. Ca ²⁺ is involved in various cellular responses, and its intracellular concentration is usually controlled within a certain range by various mechanisms, but excessive Ca ²⁺ causes contracture of cells and Ca ²⁺ Induced proteases and lipases cause cell membrane damage by activation, and further promote cell damage by reducing ATP production ability, and in severe cases, lead to cell death [circulation, (Circula)
Tion), Vol. 75, V15-V24, 1987
Year. )]. Therefore, suppressing Ca ²⁺ overload during ischemia reperfusion by inhibiting the Na ⁺ / Ca ²⁺ exchange system is considered to be effective in treating or preventing cell damage in various ischemic diseases. .

[0005] The present inventors based on the above idea, Na ⁺
As a result of diligent search for compounds having a / Ca ²⁺ exchange inhibitory effect, it was found that 2- (4-benzyloxyphenoxy) aniline and its salts satisfy the above object, and the present invention has been completed.

That is, the present invention provides

[0007]

[Formula 2]

Na ⁺ containing 2- (4-benzyloxyphenoxy) aniline and a salt thereof as an active ingredient
/ Ca ²⁺ exchange inhibitor.

In the present invention, the active ingredient represented by the formula (1)
The 2- (4-benzyloxyphenoxy) aniline represented by the formula (I) is a known compound. Chem. So
c. Chem. Commun. Volume 20, page 949.
The manufacturing method was also disclosed in 1980.

The salt may be any pharmaceutically acceptable salt, for example, an inorganic acid salt such as hydrochloride, hydrobromide or sulfate, or a methanesulfonate or p-toluenesulfonate. There are sulfonates.

[0011]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The dose of the compound which is the active ingredient of the medicament of the present invention varies depending on the symptoms, but the daily dose for an adult is usually 0.1 to 1000 m for oral administration.
g / human, once a day or divided into several times a day.

In the case of injection administration, the administration route can be appropriately selected from, for example, subcutaneous, intraperitoneal, intravenous and intramuscular injection. The dose is 0.1-100 mg / Kg per dose.

The Na ⁺ / Ca ²⁺ exchange system inhibitor of the present invention comprises
It can be used orally or parenterally by preparing it into an appropriate pharmaceutical composition (solid preparation such as tablets, pills, capsules, granules, dry syrups or injections) using an appropriately known carrier, diluent or the like. .

In order to produce a solid preparation, various additives such as excipients, disintegrants, binders, lubricants, and coating bases are used, and agitation granulation, fluidized bed granulation, crushing granulation, and the like. It can be manufactured by the method.

Examples of the excipient include mannitol, xylitol, sorbitol, glucose, sucrose, lactose, crystalline cellulose and the like.

Examples of the disintegrant include low-substituted hydroxypropylcellulose, carboxymethylcellulose, carboxymethylcellulose calcium, sodium carboxymethylcellulose and the like.

Examples of the binder include methylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, gelatin, gum arabic, ethylcellulose and the like.

Examples of the lubricant include stearic acid, magnesium stearate, calcium stearate and the like.

In addition, if necessary, an antioxidant, a coating agent, a coloring agent, a flavoring agent, a surfactant, a plasticizer and the like can be added.

In the case of administration by injection, the compound can be produced by conventional means, for example, by dissolving the compound which is the active ingredient of the present invention with alcohol, and adding physiological saline thereto to adjust the total amount.

[0021]

Industrial ^{Applicability The} present invention inhibits the Na ⁺ / Ca ²⁺ exchange system, and is thus useful for the treatment and prevention of ischemic heart disease such as myocardial infarction, ischemic brain disease such as cerebral infarction or ischemic kidney disease. It is also useful for thrombolytic therapy, angioplasty, coronary artery bypass surgery, and cell protection during organ transplantation.

[0022]

The present invention will be described below in more detail with reference to Test Examples and Examples.

Test Example 1 Inhibition of Na ⁺ / Ca ²⁺ exchange system using myocardial endoplasmic reticulum A method described in the literature (LR Jones, Methods Enzymol., 1988, 15).
7, pp85.), And membrane vesicles prepared from the isolated dog ventricular muscle were used.

The measurement of Na ⁺ / Ca ²⁺ exchange activity using membrane vesicles is carried out by the method described in the literature (KD Philipson, et al., J. Biol.
Chem., 1980, 255, pp6880.). First, membrane vesicles were treated with a sodium-containing solution [160 mM sodium chloride, 20 mM
Tris-HCl (pH 7.4)] with a protein concentration of 1.5 mg /
and suspended for 1 hour to load Na ⁺ into the membrane vesicles. Next, add 2.5 μl of the membrane vesicles to [
⁴⁵ Ca] -calcium chloride solution [20 μM [ ⁴⁵ Ca] -calcium chloride, 160 mM potassium chloride, 20 mM Mops-T
ris (pH 7.4)], and after 10 seconds, an ice-cooled lanthanum chloride solution [10 mM lanthanum chloride, 160 mM potassium chloride, 20 mM Mops-Tris (pH 7.4)] 900
μl was added, and membrane vesicles were collected on a nitrocellulose filter by suction filtration, and washed three times with 900 μl of a lanthanum chloride solution. The concentration of Ca ²⁺ incorporated into the membrane vesicles was determined by measuring ⁴⁵ Ca radioactivity with a liquid scintillator. In addition, the incorporation of Ca ²⁺ into membrane vesicles independent of Na ⁺ / Ca ²⁺ exchange activity is based on a potassium-containing solution [160 mM potassium chloride, 20 mM T
ris-hydrochloric acid (pH 7.4)] and the same operation was performed.

The test compound was a dimethyl sulfoxide solution, and its inhibitory effect was evaluated by comparing with the vehicle-treated group. The IC ₅₀ value was calculated from the dose inhibition curve of each test compound using the least squares method. Table 1 shows the results.

[0026]

[Table 1]

Test Example 2 Increased intracellular Ca ²⁺ concentration via Na ⁺ / Ca ²⁺ exchange system
The inhibitory effect on the method described in the literature (J. Sadosima, et al., J. Biol. Chem., 199
1, Vol 267, pp1051.), And cardiomyocytes were isolated and cultured from neonatal rats (SLC). That is, ventricular muscle was excised from a newborn rat, and after mins, trypsin was added to tissue slices.
A 1% solution was added and cardiomyocytes were isolated.

After preparing cardiomyocytes exhibiting automatic pulsation at 1 × 10 ⁵ cells / cm ² in a 24-well plastic petri dish, the method described in the literature (S. Wakabayashi, et al., Biochem. Biophys. A
cta, 1981, Vol 642, pp 158.).
First, a Ca ²⁺ -free solution [135 mM sodium chloride, 5 mM potassium chloride, 5.5 mM glucose, 2 mM ethylene glycol bis (2-aminoethyl ether) tetraacetic acid (EGT
A), 10 mM HEPES-Tris (pH 7.4)]
After loading the cells with Na ⁺ by perfusing at 10 ° C. for 10 minutes, a Na ⁺ -free solution [135 mM choline chloride, 5 mM
Substitution with potassium chloride, 5.5 mM glucose, 1 mM [ ⁴⁵ Ca] -calcium chloride, 10 mM HEPES-Tris (pH 7.4)] and perfusion at 37 ° C. for 1 minute to induce intracellular C
a ²⁺ was incorporated. The intracellular Ca ²⁺ concentration was determined by lysing the cells with a 0.1N aqueous sodium hydroxide solution and measuring ⁴⁵ Ca radioactivity with a liquid scintillator.

The Ca ²⁺ -free solution and the Na ⁺ -free solution were perfused in the presence of the test compound to measure the concentration of Ca ²⁺ taken up into the cells, and the concentration was suppressed by comparison with the solvent-treated group (control). The effect was evaluated. The IC ₅₀ value was calculated from the dose inhibition curve of each test compound using the least squares method. Table 2 shows the results.

[0030]

[Table 2]

Example 1 Formulation (in 1 tablet) Compound of the present invention 50 mg Lactose 40 mg Constarch 49.75 mg Crystalline cellulose 17 mg Carmellose calcium 17 mg Hydroxypropylcellulose 5.25 mg Magnesium stearate 1 mg Total 180 mg Compound of the present invention, lactose, constarch, Microcrystalline cellulose and carmellose calcium are mixed uniformly, and 1
A 0% aqueous solution of hydroxypropylcellulose was added, and the mixture was kneaded and dried. The granules were sieved with a 30M sieve to obtain uniform granules. Magnesium stearate was added, and the mixture was tabletted to give tablets.

Example 2 Formulation (in 1 tablet) Compound of the present invention 50 mg Lactose 40 mg Constarch 49.75 mg Crystalline cellulose 17 mg Carmellose calcium 17 mg Hydroxypropyl cellulose 5.25 mg Magnesium stearate 1 mg Total 180 mg Compound of the present invention, lactose, constarch, Microcrystalline cellulose and carmellose calcium are mixed uniformly, and 1
0% hydroxypropylcellulose ethanol solution was added, kneaded and dried, and the granules were sieved with a 30M sieve.
Make uniform granules, add magnesium stearate,
Tablets were made into tablets.

Example 3 Formulation (1 tablet) Compound of the present invention 50 mg Lactose 45 mg Constarch 44.75 mg Crystalline cellulose 17 mg Carmellose calcium 17 mg Hydroxypropyl cellulose 5.25 mg Magnesium stearate 1 mg Total 180 mg Compound of the present invention (CAM), lactose , Constarch, microcrystalline cellulose, carmellose calcium,
To this was added a 10% aqueous solution of hydroxypropylcellulose, kneaded and dried, and the granules were sieved with a 30M sieve.
Make uniform granules, add magnesium stearate,
Tablets were made into tablets.

Example 4 Formulation (per tablet) Compound of the present invention 50 mg Lactose 45 mg Constarch 44.75 mg Crystalline cellulose 17 mg Carmellose calcium 17 mg Hydroxypropyl cellulose 5.25 mg Magnesium stearate 1 mg Total 180 mg Compound of the present invention, lactose, constarch, crystal Cellulose and carmellose calcium are mixed uniformly, and 1
A 0% aqueous solution of hydroxypropylcellulose was added, and the mixture was kneaded and dried. The granules were sieved with a 30M sieve to obtain uniform granules. Magnesium stearate was added, and the mixture was tabletted to give tablets.

Example 5 Compound of the present invention 100 mg Lactose 500 mg Sucrose 235 mg Constarch 100 mg Carmellose sodium 10 mg Hydroxypropylcellulose 50 mg Magnesium stearate 5 mg Total 1000 mg The compound of the present invention, lactose, constarch, crystalline cellulose and carmellose calcium were uniformly mixed. Then, hydroxypropylcellulose was added, water was used as a granulating solvent, and the above mixture was subjected to fluid granulation to obtain a composition for a dry syrup.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Akiko Hamano 3- 24-1, Takada, Toshima-ku, Tokyo Taisho Pharmaceutical Co., Ltd. (72) Inventor Kazuki Tomizawa 3- 24-1, Takada, Toshima-ku, Tokyo Taisho Pharmaceutical Co., Ltd.

Claims (1)

[Claims] (1) Formula (1) A Na ⁺ / Ca ²⁺ exchange system inhibitor comprising 2- (4-benzyloxyphenoxy) aniline and a salt thereof represented by the following formula: