JP2003252794A - Apoptosis inhibitor containing aldose reductase inhibitor as active ingredient - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an aldose reductase inhibitor which prevents and/or treats apoptosis-associated diseases. <P>SOLUTION: The aldose reductase inhibitor depresses apoptosis and is effective for prevention and/or treatment of apoptosis-associated diseases, for example, dementia such as senile dementia of Alzheimer type, etc., cerebrovascular disease, neurodegenerative disease, AIDS, AIDS-related disease, HIV, HTLV- related diseases such as diseases, adult T cell leukemia, leukemic reticuloendotheliosis, myelopathia, respiratory disorder, arthropathy, uveitis, etc., collagenosis such as systemic lupus erythematosus, chronic rheumatism, etc., and ulcerous colitis, etc. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a therapeutic and / or preventive agent for apoptosis-related diseases, which contains an aldose reductase inhibitor as an active ingredient.

[0002]

BACKGROUND OF THE INVENTION AND PRIOR ART Apoptosis is characterized by rapid cell shrinkage accompanied by changes in the intranuclear structure. Chromatin loses its normal reticulated structure, aggregates at the edge of the nuclear membrane, and the entire cytoplasm including the nucleus It is a shrinking cell death. Apoptosis is thought to be deeply involved in pathological conditions such as viral infection and carcinogenesis, as well as physiological functions such as tissue formation during development, homeostasis maintenance of the living body, and aging. For example, in AIDS pathology, HIV infection induces apoptosis of CD4-positive T cells, and in cancer cells of malignant tumor patients, apoptosis is promoted by the action of cytokines from cells surrounding the cancer cells. There is a report. Therefore, having an apoptosis inhibitory action is considered to be effective for treating and preventing diseases in apoptosis-related diseases.

Examples of diseases associated with apoptosis include dementia such as Alzheimer-type senile dementia, cerebrovascular disorders, neurodegenerative diseases, AIDS, AIDS-related diseases (A
RC), adult T cell leukemia, hairy cell leukemia, myelopathy,
HIV, HTLV such as respiratory disorders, arthrosis, uveitis
Related diseases and systemic lupus erythematosus, collagen diseases such as rheumatoid arthritis, ulcerative colitis, Sjogren's syndrome, primary biliary cirrhosis, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, myasthenia gravis, insulin Dependent type (I
Type) autoimmune diseases such as diabetes, myelodysplastic syndrome, periodic thrombocytopenia, aplastic anemia, idiopathic thrombocytopenia, various diseases associated with thrombocytopenia such as generalized intravascular coagulation, C type, Liver diseases such as type A, B or F viral or drug-induced hepatitis and cirrhosis, adult respiratory distress syndrome,
Examples include benign prostatic hyperplasia, uterine fibroids, bronchial asthma, various congenital malformations, nephritis, senile cataract, chronic fatigue syndrome, muscular dystrophy and the like.

On the other hand, the aldose reductase is an enzyme that reduces aldoses in the body such as glucose and galactose to corresponding polyols such as sorbitol and galactitol. In hyperglycemic conditions such as in diabetic patients, sorbitol and galactitol produced by the excessive action of this enzyme are accumulated in the lens, peripheral nerves, kidneys, etc., and as a result, retinopathy, diabetic cataract, peripheral neuropathy,
Complications such as renal disorders occur. Aldose reductase inhibitors are known to be effective in treating and preventing complications of chronic diabetes by inhibiting aldose reductase.

The following are known examples of aldose reductase inhibitors.

Japanese Patent Application Laid-Open No. 57-40478 discloses a compound represented by the general formula (A):

[0007]

[Chemical 4]

The compound represented by has an aldose reductase inhibitory action and is caused by aldose reductase in complications of chronic diabetes such as cardiovascular disorder, renal disorder, retinopathy, diabetic cataract, neuropathy and infectious disease. Neuropathies such as neuralgia known as complications, retinopathy, diabetic cataract,
It is described to be useful for the prevention and treatment of renal disorders such as renal tubular disease.

In addition, due to the fact that pericytes are lost in diabetic retinopathy, SNK-860 [(() which has an aldose reductase inhibitory action using bovine retinal microvascular pericytes cultured under high sugar concentration conditions. 2S, 4S) -6-Fluoro-2 ′, 5′-dioxospiro [3,4-dihydro-2H-1-benzopyran-4,4′-imidazolidine] -2-carboxamide] was investigated for its effect and the result was It has been reported that the compound has an inhibitory effect on the high sugar-induced apoptosis in the cells. [Ex
p. Eye. Res., 71, 309-315 (2000)]. However, this is performed under high sugar concentration conditions, and an experiment under a stimulus different from the high sugar concentration is not known. Of course, there is no report that an aldose reductase inhibitor suppresses apoptosis under stimulating conditions different from high sugar concentration and in cells other than the above cells.

[Problems to be Solved by the Invention] A drug for treating diseases associated with apoptosis by inhibiting apoptosis by an aldose reductase inhibitor has not yet been realized.

[0010]

Means for Solving the Problems As a result of various studies on an apoptosis inhibitor, the present inventors have found that an aldose reductase inhibitor achieves this object, and completed the present invention.

That is, the present invention relates to a therapeutic and / or preventive agent for apoptosis-related diseases, which contains an aldose reductase inhibitor as an active ingredient.

Specific examples of the apoptosis-related disease in the present invention include dementia such as Alzheimer-type senile dementia, cerebrovascular disorder, neurodegenerative disease, AIDS, AIDS.
HI for related diseases (ARC), adult T cell leukemia, hairy cell leukemia, myelopathy, respiratory disorders, arthrosis, uveitis, etc.
V, HTLV-related diseases, systemic lupus erythematosus, collagen diseases such as rheumatoid arthritis, ulcerative colitis, Sjogren's syndrome, primary biliary cirrhosis, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, myasthenia gravis Disease, autoimmune diseases such as insulin-dependent (type I) diabetes, myelodysplastic syndrome, periodic thrombocytopenia, aplastic anemia, idiopathic thrombocytopenia, thrombocytopenia such as generalized intravascular coagulation Associated diseases, viral diseases such as C, A, B or F, liver diseases such as drug-induced hepatitis and cirrhosis, adult respiratory distress syndrome, benign prostatic hyperplasia, uterine fibroids, bronchial asthma, various congenital malformations , Nephritis, senile cataract, chronic fatigue syndrome, muscular dystrophy.

The apoptosis-related diseases in the present invention do not include diabetic complications, specifically diabetic retinopathy, diabetic cataract, diabetic peripheral neuropathy and diabetic nephropathy.

The aldose reductase inhibitor used in the present invention is represented by the general formula (I)

[0015]

[Chemical 5]

(In the formula, R ¹ and R ² are (1) R ² and R ² are each independently, and each of the following (i) to
A phenyl group substituted or unsubstituted by one or more groups selected from (x): (i) halogen atom, (ii) trifluoromethyl, (ii
i) hydroxyl group, (iv) nitro, (v) carboxyl group, (v
i) an amino optionally substituted with C1-4 alkyl,
(Vii) C1-4 alkyl, C1-4 alkoxy, or C1-4 alkylthio, (viii) phenyl, (ix)
Heterocycle containing at least one of nitrogen, oxygen and sulfur atoms: (The heterocycle is (a) halogen atom, (b) trifluoromethyl, (c) phenyl, (d) nitro, (e)
Hydroxyl group, (f) carboxyl group, (g) amino optionally substituted with C1-4 alkyl, (h) C1-4 alkyl, (j) C1-4 alkoxy, and (k) C1-
It may be substituted with one or more groups selected from 4 alkylthio. ), (X) a hydroxyl group, phenyl and C1-4 alkyl substituted with one or more groups selected from the heterocycles described in (ix) above; or (2) R ¹
Represents a hydrogen atom and R ² is a group represented by the following (i) to (vi): (i) C1-4 alkyl substituted with one or more C1-4 alkyl, or unsubstituted C4-7 cycloalkyl Or C4-7 cycloalkenyl, (ii) anthryl or naphthyl, (iii) a phenyl group substituted with one or more groups selected from the following (a) to (k), or an unsubstituted phenyl group: (a ) Halogen atom, (b) trifluoromethyl, (c) hydroxyl group, (d) nitro, (e) carboxyl group, (f) amino optionally substituted with C1-4 alkyl, (g) C1-4 alkyl , C1-4 alkoxy, or C1-4 alkylthio, (h) phenyl,
(J) Heterocycle containing at least one of nitrogen, oxygen and sulfur atoms: (wherein the heterocycle is (aa) halogen atom, (bb)
Trifluoromethyl, (cc) phenyl, (dd) nitro,
(Ee) hydroxyl group, (ff) carboxyl group, (gg) C1-4
Amino optionally substituted with alkyl, (hh) C1-
4-alkyl, (jj) C1-4alkoxy, and (kk)
It may be substituted with one or more groups selected from C1-4 alkylthio. ), And (k) a hydroxyl group, phenyl, and 1 selected from the heterocycles described in (j) above.
C1-4 alkyl substituted with one or more groups, (iv) substituted with one or more groups selected from the following (a) to (k), or unsubstituted nitrogen, oxygen and sulfur atoms A heterocycle containing at least one: (a) a halogen atom, (b) trifluoromethyl,
(C) phenyl, (d) nitro, (e) hydroxyl group, (f)
Carboxyl group, (g) C1-4 alkyl optionally substituted amino, (h) C1-4 alkyl, C1-4
Alkoxy or C1-4 alkylthio, (j) oxo, and (k) hydroxyl group, phenyl and the above (ii)
C1-4 alkyl substituted with one or more groups selected from the heterocycles described in (j) in i), (v) formula

[0017]

[Chemical 6]

(Wherein R ⁴ represents a hydrogen atom or C1-4 alkyl), or (vi) the formula

[0019]

[Chemical 7]

Represents a group represented by: or
(3) R ¹ and R ² together represent tetramethylene or pentamethylene, and R ³ is (1) a hydrogen atom,
(2) C1-12 alkyl, (3) C7-13 aralkyl, or (4) substituted with one or more C1-4 alkyl, or unsubstituted C4-7 cycloalkyl, or C4- 7 cycloalkenyl, or (3) phenyl substituted or unsubstituted with one or more groups selected from the following (i) to (x): (i) halogen atom, (ii) trifluoromethyl, (Ii
i) hydroxyl group, (iv) nitro, (v) carboxyl group, (v
i) an amino optionally substituted with C1-4 alkyl,
(Vii) C1-4 alkyl, C1-4 alkoxy, or C1-4 alkylthio, (viii) phenyl, (ix)
Heterocycle containing at least one of nitrogen, oxygen and sulfur atoms: (The heterocycle is (a) halogen atom, (b) trifluoromethyl, (c) phenyl, (d) nitro, (e)
Hydroxyl group, (f) carboxyl group, (g) amino optionally substituted with C1-4 alkyl, (h) C1-4 alkyl, (j) C1-4 alkoxy, and (k) C1-
It may be substituted with one or more groups selected from 4 alkylthio. ) And (x) a hydroxyl group, phenyl and C1-4 alkyl substituted with one or more groups selected from the heterocycles described in (ix) above. )
The rhodanin derivative represented by or a non-toxic salt thereof is preferable.

The compound represented by the general formula (I) can be converted into a non-toxic salt by a known method.

In the present specification, the non-toxic salts include alkali metal salts, alkaline earth metal salts, ammonium salts, amine salts,
Examples thereof include acid addition salts.

The salt is preferably non-toxic and water-soluble. Suitable salts include salts of alkali metals (potassium, sodium, etc.), salts of alkaline earth metals (calcium, magnesium, etc.), ammonium salts, pharmaceutically acceptable organic amines (tetramethylammonium, triethylamine, methylamine). , Dimethylamine, cyclopentylamine, benzylamine, phenethylamine, piperidine, monoethanolamine, diethanolamine, tris (hydroxymethyl) aminomethane, lysine, arginine, N-methyl-D-glucamine, etc.).

The acid addition salt is preferably non-toxic and water-soluble. Suitable acid addition salts include inorganic acid salts such as, for example, hydrochlorides, hydrobromides, hydroiodides, sulfates, phosphates, nitrates, or acetates, lactates, tartrates, benzoates. Organic acid salts such as acid salts, citrates, methane sulfonates, ethane sulfonates, benzene sulfonates, toluene sulfonates, isethionates, glucuronates, and gluconates.

Further, the compound of the present invention represented by the general formula (I) or a non-toxic salt thereof can be converted into a solvate by a known method.

The solvates are preferably non-toxic and water-soluble. Suitable solvates include, for example, solvates such as water and alcoholic solvents (eg ethanol).

In the present invention, all isomers are included unless otherwise specified. For example, alkyl, alkoxy and alkylthio include straight and branched ones. Furthermore, isomers (E, Z, cis, trans isomers) in double bonds, rings and condensed rings, isomers due to the presence of asymmetric carbon (R, S isomers, α isomers, β isomers, enantiomers, diastereomers) ), An optically active substance having optical activity (D, L, d, l form), a polar substance (high polar substance, low polar substance) by chromatographic separation, an equilibrium compound, a mixture of these at arbitrary ratios, and a racemic mixture , All are included in the present invention.

In the present invention, more preferred compounds are
(E, E) -5- (2-Methyl-3-phenyl-2-propenylidene) -4-oxo-2-thioxo-3-thiazolidineacetic acid (Compound (1); generic name: epalrestat).

The compound represented by the general formula (I) is
It can be produced by the method described in JP-A-57-40478.

[0030]

[Pharmacological activity of the present compound] The present compound represented by the general formula (I) has an apoptosis inhibitory action as described below, and is therefore useful for the treatment and / or prevention of the above-mentioned apoptosis-related diseases.

[0031]

[Toxicity] The toxicity of this compound was sufficiently low, and it was confirmed that the compound was sufficiently safe for use as a drug. For example, in oral administration using rats, the LD ₅₀ value of compound (1) was 5600 mg / kg.

[0032]

[Application to Pharmaceuticals] Since the aldose reductase inhibitor used in the present invention has an apoptosis-suppressing action, for example, the immune function and brain function involved in apoptosis are decreased or promoted, and circulatory functions such as arteriosclerosis and myocardial infarction. Obstacle,
It is used for the treatment and / or prevention of liver dysfunction, malignant tumor, blood cell dysfunction and the like. In particular,
Dementia such as Alzheimer-type senile dementia, cerebrovascular disorder,
Neurodegenerative diseases, AIDS, AIDS-related diseases, adult T-cell leukemia, hairy cell leukemia, myelopathy, respiratory disorders, arthritis, HIV such as uveitis, HTLV-related diseases, systemic lupus erythematosus, rheumatoid arthritis, etc. Collagen disease, ulcerative colitis, Sjogren's syndrome, primary biliary cirrhosis,
Idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, myasthenia gravis, autoimmune diseases such as insulin-dependent (type I) diabetes, myelodysplastic syndrome, periodic thrombocytopenia, aplastic anemia, Various diseases associated with thrombocytopenia such as idiopathic thrombocytopenia and generalized intravascular coagulation, liver diseases such as C-type, A-type, B-type or F-type viral or drug-induced hepatitis and cirrhosis, adults It is used for the treatment and / or prevention of respiratory distress syndrome, benign prostatic hyperplasia, uterine myoma, bronchial asthma, various congenital malformations, nephritis, senile cataract, chronic fatigue syndrome, muscular dystrophy and the like.

The aldose reductase inhibitor used in the present invention comprises: 1) complementation and / or enhancement of the preventive and / or therapeutic effect of the inhibitor on diseases related to apoptosis, 2) improvement of the kinetics / absorption of the inhibitor, dose And / or 3) it may be administered as a concomitant drug in combination with other drugs in order to reduce the side effects of the inhibitor.

The combination drug of the aldose reductase inhibitor and the other drug may be administered in the form of a preparation in which both components are mixed in one preparation, or in the form of separate preparations for administration. . When administered as separate formulations, simultaneous administration and administration with a time difference are included. In addition, the administration by the time difference may be performed by first administering the aldose reductase inhibitor and then administering the other drug, or by administering the other drug first and subsequently administering the aldose reductase inhibitor. Regardless, the respective administration methods may be the same or different.

The disease which exerts a preventive and / or therapeutic effect by the above-mentioned combination drug is not particularly limited, and it is a disease which complements and / or enhances the preventive and / or therapeutic effect of the apoptosis-related disease of the aldose reductase inhibitor. Good.

For example, other drugs for complementing and / or enhancing the preventive and / or therapeutic effect of aldose reductase inhibitor on dementia such as Alzheimer-type senile dementia include, for example, acetylcholinesterase inhibitor and nicotine. Receptor modulators, cerebral circulation metabolism improvers, monoamine oxidase inhibitors, vitamin E and the like can be mentioned.

[0037] For example, other agents for complementing and / or enhancing the preventive and / or therapeutic effect of aldose reductase inhibitor on cerebrovascular disorder include, for example, β
Blocking agents, antithrombotic agents, cerebral circulation metabolism improving agents, etc.

[0038] For example, other agents for complementing and / or enhancing the preventive and / or therapeutic effect of an aldose reductase inhibitor on neurodegenerative diseases include, for example,
Antiepileptic drugs, acetylcholinesterase inhibitors, astrocyte function improving agents, neurotrophic factors and the like can be mentioned.

Examples of the acetylcholinesterase inhibitor include donevezil hydrochloride, TAK-147 and the like.

Examples of the cerebral circulatory metabolism improving agents include cerebral vasodilators, cerebral metabolism activating agents, blood property improving agents, and the like. Specifically, idebenone, calcium fopantenate, amantadine hydrochloride, meclofenoki hydrochloride. Sart, dihydroergotoxin mesylate, pyrithioxine hydrochloride,
Examples include γ-aminobutyric acid, bifemelane hydrochloride, lisuride maleate, inderoxazine hydrochloride, nicergoline, propentofylline and the like.

As the monoamine oxidase inhibitor,
Safradine hydrochloride may be mentioned.

Examples of β-blockers include atenolol, nadolol, nipradilol, pindolol, bisoprolol fumarate, timolol maleate, popindolol malonate, acebutolol hydrochloride, alprenolol hydrochloride, indenolol hydrochloride, oxprenolol hydrochloride, carte hydrochloride. Orol, Ceriprolol Hydrochloride, Chilisolol Hydrochloride, Bucumolol Hydrochloride, Bufetrol Hydrochloride,
Examples thereof include bupranolol hydrochloride, propranolol hydrochloride, betaxolol hydrochloride, metoprolol tartrate, and penbutolol sulfate.

Examples of antithrombotic agents include heparin, oral anticoagulants (warfarin), synthetic antithrombin agents (gavexate mesylate, nafamostat mesylate, argatroban, etc.), antiplatelet agents (aspirin, dipyridamole, ticlopine hydrochloride, Beraprost sodium, cilostazol, ozagrel sodium, etc.), thrombolytic agents (urokinase, tisokinase, alteplase, etc.),
Factor Xa inhibitors and Factor VIIa inhibitors can be mentioned.

As the antiepileptic drug, phenobarbital, mehobarbital, metalbital, primidone,
Phenytoin, ethotoin, trimetadione, ethosuximide, acetylphenetride, carbamazepine, acetazolamide, diazepam, sodium valproate and the like can be mentioned.

ON as an astrocyte function improving agent
O-2506 is mentioned.

Examples of the neurotrophic factor include ABS-205.

The weight ratio of the compound represented by the general formula (I) to the other drug is not particularly limited.

The other drugs may be administered in combination of any two or more kinds.

Further, other drugs that complement and / or enhance the preventive and / or therapeutic effect of apoptosis-related diseases by aldose reductase inhibitors have been found to date based on the above-mentioned mechanism. Not only that, but also those found in the future are included.

In order to use the combination agent of the aldose reductase inhibitor used in the present invention and another drug for the above purpose, usually,
It is administered systemically or locally, orally or parenterally.

The dose depends on age, body weight, symptoms, therapeutic effect,
Although it varies depending on the administration method, treatment time, etc., it is usually 1 dose per adult per dose in the range of 1 mg to 1000 mg.
Orally administered once to several times a day, or once per adult, in the range of 1 mg to 100 mg, parenterally administered once to several times a day (preferably intravenous administration), Alternatively, it is continuously administered intravenously within the range of 1 hour to 24 hours per day.

Of course, as described above, since the dose varies depending on various conditions, a dose smaller than the above dose may be sufficient in some cases, or a dose exceeding the range may be necessary.

When administering a combination of an aldose reductase inhibitor and another drug, a solid composition for oral administration,
It is used as a liquid composition and other compositions, as an injection for parenteral administration, an external preparation, a suppository, an eye drop, an inhalant and the like.

Solid compositions for oral administration include tablets,
Pills, capsules, powders, granules and the like are included.

Capsules include hard capsules and soft capsules.

In such solid compositions, the one or more active substances comprises at least one inert diluent such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone, Mixed with magnesium aluminometasilicate. The composition comprises, according to a conventional method, additives other than an inert diluent, for example, a lubricant such as magnesium stearate, a disintegrant such as calcium fibrin glycolate, a stabilizer such as lactose, glutamic acid or asparagine. It may contain a solubilizing agent such as an acid. If necessary, the tablets or pills may be coated with a film of a gastric or enteric substance such as sucrose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose phthalate, or may be coated with two or more layers. . Also included are capsules of absorbable material such as gelatin.

Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, syrups, elixirs and the like. In such a liquid composition, one or more active substances are contained in a commonly used inert diluent (eg, purified water, ethanol). The composition may contain, in addition to an inert diluent, auxiliary agents such as a wetting agent and a suspending agent, a sweetening agent, a flavoring agent, an aromatic agent, and a preservative.

Other compositions for oral administration include spray formulations which contain one or more active substances and are formulated in a manner known per se. This composition contains, in addition to an inert diluent, a stabilizer such as sodium bisulfite and a buffer that provides isotonicity, for example, isotonic agents such as sodium chloride, sodium citrate or citric acid. May be. The manufacturing method of the spray agent is, for example, U.S. Pat.
It is described in detail in No. 95,355.

The injections for parenteral administration according to the present invention include sterile aqueous and / or non-aqueous solutions, suspensions and emulsions. Examples of the aqueous solution and suspension include distilled water for injection and physiological saline.
Examples of the water-insoluble solution and suspension include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, Polysorbate 80 (registered trademark) and the like. Further, aseptic aqueous and non-aqueous solutions, suspensions and emulsions may be mixed and used. Such compositions may further contain adjuvants such as preservatives, wetting agents, emulsifiers, dispersants, stabilizers (eg lactose), solubilizers (eg glutamic acid, aspartic acid). . These are sterilized by filtration through a bacteria-retaining filter, blending of a bactericide, or irradiation. They can also be used by producing a sterile solid composition, for example, by dissolving in sterilized or sterile distilled water for injection or other solvent before use of a lyophilized product.

Dosage forms of eye drops for parenteral administration include eye drops, suspension type eye drops, emulsion type eye drops, reconstituted eye drops and eye ointment.

These eye drops are manufactured according to known methods. For example, in the case of eye drops, a tonicity agent (sodium chloride, concentrated glycerin, etc.), a buffering agent (sodium phosphate, sodium acetate, etc.), a surfactant (Polysorbate 80 (trade name), polyoxyl stearate 40) , Polyoxyethylene hydrogenated castor oil, etc.), stabilizers (sodium citrate, sodium edetate, etc.), preservatives (benzalkonium chloride, paraben, etc.) and the like are appropriately selected and prepared. These are sterilized in the final step or prepared by an aseptic method.

Inhalants for parenteral administration include aerosols, powders for inhalation or liquids for inhalation, which are dissolved or suspended in water or other appropriate medium at the time of use. It may be used.

These inhalants are manufactured according to known methods.

For example, in the case of inhalation liquids, preservatives (benzalkonium chloride, paraben, etc.), coloring agents, buffering agents (sodium phosphate, sodium acetate, etc.), isotonic agents (sodium chloride, concentrated, etc.) Glycerin and the like), a thickener (carboxyvinyl polymer and the like), an absorption promoter and the like are appropriately selected and prepared.

In the case of powder for inhalation, lubricants (stearic acid and salts thereof, etc.), binders (starch, dextrin etc.), excipients (lactose, cellulose etc.), colorants, preservatives (chloride) (Benzalkonium, paraben, etc.), an absorption promoter, etc. are appropriately selected and prepared as necessary.

A sprayer (atomizer, nebulizer) is usually used when administering the liquid preparation for inhalation, and an inhaler administration device for powder medicine is usually used when administering the powder preparation for inhalation.

Other compositions for parenteral administration include external solutions, ointments, coatings, suppositories for rectal administration, which contain one or more active substances and are prescribed by a conventional method. Includes pessaries and the like for vaginal administration.

[0068]

[Examples] It was confirmed by the following experiments that the aldose reductase inhibitor compound has an apoptosis inhibitory action.

Compound (1) used as a test drug
Is (E, E) -5- (2-methyl-3-phenyl-2
-Propenylidene) -4-oxo-2-thioxo-3-
Thiazolidine acetic acid.Example 1: Apoptosis of serum-free culture of IMR-32 cells
Sith (1) Preparation of IMR-32 cells and drug treatment IMR-32 cells were treated with 10% fetal bovine serum (hereinafter, FB
Abbreviated as S) containing Dulbecco's modified Eagle medium (DME
3 x 10 in M)⁵Suspended to cells / mL,
3x10 in 96 well flat bottom plate^Fourcells / we
11/100 μL was added (vehicle group). Well
Separately, 3x10 was added to serum-free DMEM. ⁵cell
Suspend the cells at s / mL and add 96-well flat bottom plate.
3x10 on the board^FourAddition of cells / well / 100μL
(Serum-free group). In addition, the final serum-free DMEM
Test compound (1) was added to a concentration of 30 μM,
3x10 cells⁵Suspend at cells / mL
3x10 in a 96-well flat bottom plate^Fourcells /
well / 100 μL was added (compound (1) group).

After seeding the cells, the cells were cultured for 9 to 96 hours, and TUNEL staining (5) described below was performed for the detection of apoptotic cells. Example 2: High glutamate in rat dorsal root ganglion cells
Apoptosis upon stimulation (2) Schwann cell preparation using rat dorsal root ganglia and drug treatment From neonatal Wistar rats, dorsal root ganglia (hereinafter,
Abbreviated as DRG. ), 0.25% per 5 animals
Treat with collagenase (3 mL) at 37 ° C for 30 minutes,
1 μL DNase I-containing trypsin / EDTA (3
The cells were dispersed by suspending the cells at 37 ° C for 15 minutes. Then, an equal amount of DMEM / 10% F
The cells were washed by inactivating trypsin with BS medium. D
Suspended in MEM / 10% FBS, 1.5x10 ⁵ cells
3 × 10 ⁴ cells / well / 200 μL was seeded on an ls / mL polylysine-coated 96-well flat bottom plate, and cultured at 37 ° C. for 1 day. Next, the culture solution was removed by suction, and DMEM / 10% FBS (vehicle)
Group), and a culture medium (compound (1) group) in which DMEM / 10% FBS was added to a final concentration of 25 mM glutamic acid-added culture solution (high-glutamic acid group) and a final concentration of 30 μM was added to compound (1). did. 24 hours to 7
After culturing for 2 hours, (5) TUNEL staining described below was performed for the detection of apoptotic cells. Example 3: A when PC-12 cells are stimulated with high glutamate
Apoptosis (3) Preparation and drug treated PC-12 cells cells PC-12 cells, were suspended so as to be 3x10 ⁵ cells / mL in 5% FBS and 10% equine serum-containing DMEM, 3x10 ⁴ to 96-well flat-bottom plates cell
s / well / 100 μL was added (vehicle
group). Separately, the cells were suspended in DMEM containing 5% FBS and 10% horse serum at 3 × 10 ⁵ cells / mL, and the cells were suspended in a 96-well flat bottom plate at 3 × 10 ⁴ cells.
Cells / well / 100 μL were added, and finally glutamic acid was added so that the final concentration was 25 mM (high glutamic acid group). Separately, 5% FBS and 10
Compound (1) was added to DMEM containing 10% horse serum to a final concentration of 30 μM, and the cells were added to 3 × 10 ⁵ cells.
/ ML, and add 3 × 10 ⁴ cells / well / 100 μL to a 96-well flat bottom plate,
Finally, glutamic acid was added so that the final concentration was 25 mM (compound (1) group).

After inoculation of the cells, the cells were cultured for 9 to 96 hours, and the following (5) TUNEL staining was carried out to detect apoptotic cells. Example 4: Apoptosis of HL-60 cells upon TNF-α stimulation
(4) Preparation of HL-60 cells 3 ml of HL-60 cells was added to RPMI medium containing 10% FBS.
Suspend at 10 ⁵ cells / mL and add 3 × 10 ⁴ cells / well / 1 to a 96-well flat bottom plate.
00 μL was added (vehicle group). Similarly, 10% FBS-containing RPMI medium was added to 3 × 10 ⁵ cells
The cells are suspended to give a final concentration of 1 ng / mL, and 3x10 ⁴ cells / well / 100 µL are added to a 96-well flat bottom plate.
-Α was added (TNF-α group). Apart from this, 10
Compound (1) was added to RPMI medium containing% FBS so that the final concentration was 30 μM, and 3 × 10 ⁵ cells / mL was added.
Cells in a 96-well flat bottom plate
x10 ⁴ cells / well / 100 μL was added.
After that, human T was further adjusted to a final concentration of 1 ng / mL.
NF-α was added (compound (1) group).

After culturing for 9 hours, (5) TUNEL staining described below was carried out for the detection of apoptotic cells. (5) TUNEL staining method To detect and quantify apoptotic cells, TUNEL
Staining was used.

That is, after each culture time of the above (1) to (4), centrifugation was performed at 1500 rpm for 5 minutes, and the supernatant was removed with a micropipette so as not to aspirate the cells. Then -8
Freeze the cells at 0 ° C for 15 minutes, and before thawing,
The cells were fixed with μL of cell fixative and centrifuged at 1500 rpm for 5 minutes to remove the supernatant. Then, treat with 100 μL of cell membrane permeate, immediately centrifuge at 1500 rpm for 5 minutes, and then on ice.
After leaving for 10 minutes, the supernatant was removed. Then, the plate was washed twice with 200 μL of PBS (−), and after removing the supernatant, 50 μL of T
The dT solution was added and the mixture was allowed to stand in a 37 ° C CO2 incubator. Wash twice with 200 μL PBS (-), 1
Add 00 μL of H2O2 and let stand for 5 minutes. 200 μL
Wash with PBS (-) 3 times, 100 μL of Perox
Idase-conjugated-anti-TdT antibody was added and the mixture was allowed to stand in a 37 ° C CO2 incubator.
Then, wash 5 times with 200 μL of PBS (−),
L of Peroxidase substrate was added, and after adequate color development, the reaction was stopped with 100 μL of HCl solution. Substrate color development using an immunoplate reader, 490 nm-650 n
It was measured in m. The obtained results are shown as OD values. (1) Results of Example 1 Compound (1) suppressed apoptosis induced in IMR-32 cells 9 hours and 48 hours after the start of serum-free culture. The results are shown in FIGS. 1 and 2. (2) Results of Example 2 48 hours after the start of stimulation with high glutamate, compound (1) suppressed apoptosis induced in rat DRG cells. The results are shown in Fig. 3. (3) Results of Example 3 Compound (1) suppressed the apoptosis induced in PC-12 cells 24 hours after the start of stimulation with high glutamate. The results are shown in Fig. 4. (4) Results of Example 4 9 hours after the start of TNF-α stimulation, compound (1) suppressed the apoptosis induced in HL-60 cells.
Results are shown in FIG. Formulation Example Formulation Example 1 The following components were admixed in a conventional method and punched out to give 100 tablets each containing 50 mg of active ingredient.・ (E, E) -5- (2-Methyl-3-phenyl-2-propenylidene) -4-oxo-2-thioxo-3-thiazolidineacetic acid ...... 5.0 g ・ Carboxymethylcellulose calcium (disintegrant) ...... 0.2 g Magnesium stearate (lubricant) ...... 0.1 g Microcrystalline cellulose ...... 4.7 g Formulation example 2 The following components were mixed by a conventional method, the solution was sterilized by a conventional method, and each 5 ml was filled in an ampoule. Lyophilization was carried out by a conventional method to obtain 100 ampules containing 20 mg of active ingredient in 1 ampule.・ (E, E) -5- (2-Methyl-3-phenyl-2-propenylidene) -4-oxo-2-thioxo-3-thiazolidineacetic acid ...... 2.0 g ・ Mannitol ...... 20 g ・ Distilled water ...... 500 ml

[Brief description of drawings]

FIG. 1 shows apoptosis induced when IMR-32 cells were serum-free cultured for 9 hours, and the inhibitory effect of compound (1) on the apoptosis.

FIG. 2 shows apoptosis induced when IMR-32 cells were serum-free cultured for 48 hours, and the inhibitory effect of compound (1) on the apoptosis.

FIG. 3 shows apoptosis induced when rat DRG cells were stimulated with high glutamate for 48 hours, and the inhibitory effect of compound (1) on the apoptosis.

FIG. 4 shows apoptosis induced when PC-12 cells were stimulated with high glutamate for 24 hours, and the inhibitory effect of compound (1) on the apoptosis.

FIG. 5 shows apoptosis induced when HL-60 cells were stimulated with TNF-α for 9 hours, and the inhibitory effect of compound (1) on the apoptosis.

Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 7/00 A61P 7/00 7/06 7/06 11/00 11/00 11/06 11/06 13/12 13 / 12 15/00 15/00 19/02 19/02 21/00 21/00 25/00 25/00 25/28 25/28 27/12 27/12 29/00 101 29/00 101 31/12 31 / 12 31/18 31/18 35/02 35/02 37/02 37/02 43/00 111 43/00 111 C07D 277/36 C07D 277/36 (72) Inventor Atsuto Inoue Shimamoto-cho, Mishima-gun, Osaka Prefecture Sakurai 3-chome No. 1-1 Ono Pharmaceutical Co., Ltd. Minase Research Institute F-term (reference) 4C033 AD01 AD02 AD16 AD20 4C084 AA17 ZA012 ZA152 ZA162 ZA332 ZA512 ZA552 ZA592 ZA662 ZA752 ZA812 ZB152 ZB12 ZB152 ZB152 ZB152 ZB152 ZB152 ZB152 ZB152 ZB272 ZB272 ZB272 ZB272 ZB272 ZB272 ZB272 ZB272 ZB272 ZB152 BC82 MA01 MA04 ZA01 ZA15 ZA16 ZA33 ZA51 ZA54 ZA55 ZA59 ZA66 ZA75 ZA81 ZA94 ZA96 ZB07 ZB15 ZB27 ZB33 ZC20 ZC35 ZC55

Claims (4)

[Claims] 1. A therapeutic and / or preventive agent for apoptosis-related diseases, which comprises an aldose reductase inhibitor as an active ingredient. 2. An aldose reductase inhibitor has the general formula (I): (In the formula, R ¹ and R ² are (1) R ² and R ² are each independently substituted with one or more groups selected from the following (i) to (x), or Unsubstituted phenyl group: (i) halogen atom, (ii) trifluoromethyl, (ii
i) hydroxyl group, (iv) nitro, (v) carboxyl group, (v
i) an amino optionally substituted with C1-4 alkyl,
(Vii) C1-4 alkyl, C1-4 alkoxy, or C1-4 alkylthio, (viii) phenyl, (ix)
Heterocycle containing at least one of nitrogen, oxygen and sulfur atoms: (The heterocycle is (a) halogen atom, (b) trifluoromethyl, (c) phenyl, (d) nitro, (e)
Hydroxyl group, (f) carboxyl group, (g) amino optionally substituted with C1-4 alkyl, (h) C1-4 alkyl, (j) C1-4 alkoxy, and (k) C1-
It may be substituted with one or more groups selected from 4 alkylthio. ), (X) a hydroxyl group, phenyl and C1-4 alkyl substituted with one or more groups selected from the heterocycles described in (ix) above; or (2) R ¹
Represents a hydrogen atom and R ² is a group represented by the following (i) to (vi): (i) C1-4 alkyl substituted with one or more C1-4 alkyl, or unsubstituted C4-7 cycloalkyl Or C4-7 cycloalkenyl, (ii) anthryl or naphthyl, (iii) a phenyl group substituted or unsubstituted with one or more groups selected from the following (a) to (k): (a ) Halogen atom, (b) trifluoromethyl,
(C) hydroxyl group, (d) nitro, (e) carboxyl group,
(F) amino optionally substituted with C1-4 alkyl, (g) C1-4 alkyl, C1-4 alkoxy, or C1-4 alkylthio, (h) phenyl, (j)
Heterocycle containing at least one of nitrogen, oxygen and sulfur atoms: (The heterocycle is (aa) halogen atom, (bb) trifluoromethyl, (cc) phenyl, (dd) nitro, (ee)
Hydroxyl group, (ff) carboxyl group, (gg) C1-4 alkyl optionally substituted amino, (hh) C1-4 alkyl, (jj) C1-4 alkoxy, and (kk) C1
It may be substituted with one or more groups selected from 4 alkylthio. ), And (k) a hydroxyl group, phenyl and C1-4 alkyl substituted with one or more groups selected from the heterocycles described in (j) above, (iv) (a) below.
A heterocycle substituted with one or more groups selected from (k) or containing at least one of unsubstituted nitrogen, oxygen and sulfur atoms: (a) halogen atom, (b)
Trifluoromethyl, (c) phenyl, (d) nitro,
(E) hydroxyl group, (f) carboxyl group, (g) C1-4
Amino optionally substituted with alkyl, (h) C1-
One or more groups selected from 4-alkyl, C1-4 alkoxy, or C1-4 alkylthio, (j) oxo, and (k) hydroxyl group, phenyl, and the heterocycle described in (j) in (iii) above. C1-4 alkyl substituted with, (v) formula (In the formula, R ⁴ represents a hydrogen atom or C1-4 alkyl), or (vi) a compound represented by the formula: Or represents (3) R ¹ and R ²
Together represent tetramethylene or pentamethylene, and R ³ is (1) a hydrogen atom, (2) C 1-12.
Or (3) C7-13 aralkyl, or (4) one or more C1-4 alkyl-substituted or unsubstituted C4-7 cycloalkyl, or C4-7 cycloalkenyl, or ( 3) Phenyl substituted or unsubstituted by one or more groups selected from the following (i) to (x): (i) halogen atom, (ii) trifluoromethyl, (iii) hydroxyl group, ( i
v) nitro, (v) carboxyl group, (vi) C1-4 alkyl optionally substituted alkyl, (vii) C1-4
A heterocycle containing at least one of alkyl, C1-4 alkoxy, or C1-4 alkylthio, (viii) phenyl, (ix) nitrogen, oxygen and sulfur atoms: (wherein the heterocycle is (a) halogen atom, (b) ) Trifluoromethyl,
(C) phenyl, (d) nitro, (e) hydroxyl group, (f)
Carboxyl group, (g) C1-4 alkyl optionally substituted amino, (h) C1-4 alkyl, (j) C
It may be substituted with one or more groups selected from 1-4 alkoxy and (k) C1-4 alkylthio. ), And (x) hydroxyl group, phenyl and the above (i
C1-4 alkyl substituted with one or more groups selected from the heterocycles described under x). The therapeutic and / or prophylactic agent according to claim 1, which is a rhodanin derivative represented by the formula (4) or a non-toxic salt thereof. 3. The aldose reductase inhibitor is (E,
E) -5- (2-Methyl-3-phenyl-2-propenylidene) -4-oxo-2-thioxo-3-thiazolidineacetic acid, The therapeutic and / or prophylactic agent according to claim 2. 4. The apoptosis-related disease is dementia such as Alzheimer-type senile dementia, cerebrovascular disorder, neurodegenerative disease, AIDS, AIDS-related disease, adult T-cell leukemia,
HIV such as hairy cell leukemia, myelopathy, respiratory disorders, arthrosis, uveitis, HTLV-related diseases and systemic lupus erythematosus, collagen diseases such as rheumatoid arthritis, ulcerative colitis,
Sjogren's syndrome, primary biliary cirrhosis, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, myasthenia gravis, autoimmune diseases such as insulin-dependent (type I) diabetes, myelodysplastic syndrome, periodicity Various diseases associated with thrombocytopenia such as thrombocytopenia, aplastic anemia, idiopathic thrombocytopenia, generalized intravascular coagulation, C type, A type, B type or F
In liver diseases such as viral and drug-induced hepatitis and liver cirrhosis, adult respiratory distress syndrome, benign prostatic hyperplasia, uterine fibroids, bronchial asthma, various congenital malformations, nephritis, senile cataract, chronic fatigue syndrome, muscular dystrophy The therapeutic and / or prophylactic agent according to claim 1.