JPH08239322A - Medicine composition containing pyrrolidinone derivative - Google Patents

Abstract

PURPOSE: To obtain a highly safe therapeutic medicine containing a specific pyrrolidinone derivative as an active ingredient, having an interleukin-1β production-inhibiting action and a tumor-necrotizing factor α release-inhibiting action, and effective for diseases such as chronic articular rheumatism and ichorrhemia. CONSTITUTION: An interleukin-1β production-inhibiting medicine and a tumor- necrotizing factor α release-inhibiting medicine contains each a pyrrolidinone derivative of the formula (R is phenyl or pyridyl) or its pharmacologically acceptable salt as an active ingredient. The pyrrolidinone derivative is low in toxicity and useful for preventing or treating diseases such as chronic articular rheumatism, rheumatic polyarthritis, ichorrhemia, systemic erythematoides, systemic scleroderma, Behcet disease, etc., The pyrrolidinone derivative includes 1-cinnamoyl-2-pyrrolidinone. The administration amount of the pyrrolidinone derivative for the expression of its activity is usually 5mg/d to 6g/d for an adult. The compound is preferably orally, intraarticularly or intravenously administered.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an interleukin-1β (hereinafter referred to as “IL-1β”) production inhibitor containing a pyrrolidinone derivative as an active ingredient, and tumor necrosis factor α (hereinafter referred to as “TNFα”) release. Regarding inhibitors.

[0002]

2. Description of the Related Art IL-1β is a protein mainly produced by immunocompetent cells such as macrophages and neutrophils, and is an important factor of immune response. Furthermore, as a factor that plays a central role during inflammation, hematopoiesis, endocrine,
It is known as a factor related to many biological reactions such as nerves. For example, in recent years, the relationship with inflammatory diseases such as rheumatoid arthritis has been clarified, IL-1β is detected in the synovium of patients with rheumatoid arthritis, and IL in synovial fluid is detected.
It has also been reported that -1β concentration correlates with local inflammatory findings.

Further, the following general formula (I)

Embedded image [Wherein, R represents a phenyl group which is unsubstituted or substituted by a lower alkoxy group or a nitro group, or a pyridyl group] is a plant growth regulator,
It is known as a methane fermentation accelerator, a cerebral circulatory metabolism improving agent and the like (Japanese Patent Laid-Open Nos. 61-229801 and 61-61).
230798, JP-A-2-225458, etc.)
However, its IL-1β production inhibitory action and TNF release inhibitory action have not been known so far.

[0004]

At present, steroid agents and non-steroidal anti-inflammatory agents are used for the treatment of inflammatory diseases such as rheumatoid arthritis. Steroids remarkably improve various symptoms in various inflammatory diseases, but with prolonged administration, resistance develops, and there are sometimes serious side effects such as digestive disorders, skin disorders and nephritis. Is a problem. On the other hand, non-steroidal anti-inflammatory agents temporarily suppress inflammatory symptoms, but do not fundamentally treat inflammatory diseases. Under such circumstances, some compounds having an inhibitory action on IL-1β production have been proposed as therapeutic agents for inflammatory diseases, but no sufficiently satisfactory compounds have been obtained. So IL-
A compound that strongly suppresses the production of 1β and has high safety has been demanded.

[0005]

Means for Solving the Problems As a result of earnest studies, the present inventors have found that the above-mentioned pyrrolidinone derivative has an excellent IL-1β production inhibitory action and a TNFα release inhibitory action, and a therapeutic agent for rheumatoid arthritis, The present invention has been completed by discovering that it is useful as a therapeutic agent for sepsis.

That is, the present invention provides the following general formula (I)

Embedded image [Wherein R represents a phenyl group which is unsubstituted or substituted by a lower alkoxy group or a nitro group, or a pyridyl group] or a pyrrolidinone derivative or a pharmaceutically acceptable salt thereof is contained as an active ingredient. , IL-1β
The present invention relates to a production inhibitor and a TNFα release inhibitor, which are useful as therapeutic agents for rheumatoid arthritis and sepsis.

In the phenyl group substituted with a lower alkoxy group represented by R in the above general formula, the lower alkoxy group is derived from a straight chain or branched chain alkyl having 1 to 4 carbon atoms, for example, a methoxy group. , Ethoxy group,
An n-propoxy group, an i-propoxy group, an n-butoxy group, an i-butoxy group, a tert-butoxy group and the like are included.

Examples of the pyrrolidinone derivative represented by the general formula (I) include the compounds shown below. 1-cinnamoyl-2-pyrrolidinone (hereinafter referred to as "compound 1"), 1- (o-methoxy-cinnamoyl) -2-pyrrolidinone (hereinafter referred to as "compound 2"), 1- (m-methoxy-cinnamoyl)- 2-pyrrolidinone (hereinafter referred to as "compound 3"), 1- (p-methoxy-cinnamoyl) -2-pyrrolidinone (hereinafter, referred to as "compound 3")
"Compound 4"), 1- (3,4-dimethoxy-cinnamoyl) -2-pyrrolidinone (hereinafter "Compound 5")
, 1- (3,4,5-tricinnamoyl) -2-
Pyrrolidinone (hereinafter referred to as "Compound 6"), 1- (o
-Nitro-cinnamoyl) -2-pyrrolidinone (hereinafter,
"Compound 7"), 1- (m-nitro-cinnamoyl) -2-pyrrolidinone (hereinafter "compound 8"), 1- (p-nitro-cinnamoyl) -2-pyrrolidinone (hereinafter "compound 9") , 1- (2,4-
Dinitro-cinnamoyl) -2-pyrrolidinone "hereinafter,
"Compound 10"), 1- (2,4,6-trinitro-cinnamoyl) -2-pyrrolidinone (hereinafter referred to as "Compound 11"), 1- (3- (2-pyridyl) -acryloyl) -2- Pyrrolidinone (hereinafter referred to as "Compound 12"), 1- (3- (3-pyridyl) -acryloyl)
-2-pyrrolidinone (hereinafter referred to as "compound 13"),
1- (3- (4-pyridyl) -acryloyl) -2-pyrrolidinone (hereinafter referred to as "compound 14"). The structures of these pyrrolidinone derivatives are shown below.

[0009]

[Table 1]

The pyrrolidinone derivative represented by the above general formula (I) can be used as desired after being converted into a salt with a pharmacologically acceptable acid, and IL-1β production inhibition containing these salts is suppressed. Agents and TNFα release inhibitors are also included in the present invention. Examples of the pharmaceutically acceptable acid include salts with mineral acids such as hydrochloric acid, sulfuric acid, hydrobromic acid and phosphoric acid; organic sulfonic acids such as methanesulfonic acid, benzenesulfonic acid and p-toluenesulfonic acid; Acetic acid, propionic acid, oxalic acid, succinic acid, maleic acid,
Examples thereof include salts with organic carboxylic acids such as fumaric acid, lactic acid, tartaric acid, malic acid and citric acid.

The pyrrolidinone derivative represented by the general formula (I) having the IL-1β production inhibitory action and the TNFα release inhibitory action of the present invention can be produced by various methods depending on the kind. Further, the salt of the pyrrolidinone derivative represented by the general formula (I) can be produced by an ordinary salt forming reaction. This pyrrolidinone derivative has excellent IL-1β production inhibitory action and is low in toxicity, therefore, rheumatoid arthritis, rheumatoid polyarthritis,
Sepsis, systemic lupus erythematosus, systemic scleroderma, Behcet's disease, periarteritis nodosa, ulcerative colitis, active chronic hepatitis, glomerulonephritis, osteoarthritis, gout, atherosclerosis, psoriasis, atopic skin I for inflammation, osteoporosis, etc.
It is useful for prevention or treatment of diseases associated with L-1β.

Further, this pyrrolidinone derivative is TNFα
Having a release-suppressing action, it is useful for the prevention or treatment of diseases in which TNFα is involved in the progression of pathological conditions. These diseases include, for example, the above-mentioned ILs such as rheumatoid arthritis, rheumatoid polyarthritis, sepsis and atopic dermatitis.
Diseases that also involve -1β, as well as acquired immune deficiency syndrome (AIDS) and the like can be mentioned.

The dose of the pyrrolidinone derivative for effective expression of the activity is usually 5 mg to 6 g, preferably 10 mg to 300 mg per adult per day.
Examples of the administration method of this compound include oral administration, intravenous administration, subcutaneous administration, intramuscular administration, rectal administration, intraarticular administration and the like, but oral administration, intraarticular administration and intravenous administration are preferred. The pyrrolidinone derivative can be prepared for administration using any conventional formulation method. Examples of the orally administered agent include tablets, granules, powders, hard capsules, soft capsules, oral liquid preparations, and the like.

Tablets and capsules used for oral administration include binders such as dextrin, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone and macrogol; excipients such as lactose, corn starch, Calcium phosphate, magnesium aluminometasilicate and the like; lubricants such as calcium stearate, talc and the like; disintegrating agents such as carboxymethyl cellulose, crystalline cellulose and the like may be contained.
Also, these formulations may be coated by methods well known in the art.

Liquid oral preparations may be aqueous or oily suspensions, emulsions, solutions, syrups, elixirs, etc. or are reconstituted with water or other suitable vehicle before use. It may be a dry product.
Such liquid formulations include commonly used additives such as suspending agents such as sorbit syrup, carboxymethyl cellulose, gelatin, hydroxyethyl cellulose, aluminum stearate gel, hydrogenated edible oils and fats; emulsifiers such as lecithin, Monooleic acid glyceride, gum arabic and the like; water-insoluble vehicle such as palm oil, propylene glycol, ethyl alcohol and the like; preservatives such as ethyl p-hydroxybenzoate, sorbic acid and the like may be contained. Examples of the dosage form for parenteral administration include injections and suppositories. The injection is prepared by a conventional method by adding a pH adjusting agent, a buffering agent, a stabilizing agent, a preservative, a solubilizing agent, etc., if necessary.

The present invention will be specifically described below with reference to examples. The present invention is not limited to these examples.

[0017]

【Example】

Example 1 (Measurement of IL-1β production inhibitory action) The IL-1β production inhibitory action of the pyrrolidinone derivative of the present invention was measured according to the following method. In RPMI 1640 medium (manufactured by Bio Whitker), 10 v / v% fetal bovine serum, 2 mM glutamine, 50 μM 2-mercaptoethanol, 60 μg / ml penicillin and 100 μm.
g / ml streptomycin was added and mixed, to which
Human peripheral blood-derived THP-1 cells (ATCC TIB2
02) is added and suspended. This suspension of THP-1 cells is subcultured at 37 ° C. under an atmosphere of a mixed gas of 95% oxygen and 5% carbon dioxide. A part of this subcultured suspension is centrifuged at 1200 rpm for 3 minutes at room temperature to recover the subcultured THP-1 cells.

Then, in RPMI 1640 medium, 2 v / v% fetal bovine serum, 2 mM glutamine, 50 μM 2-mercaptoethanol, 60 μg / ml penicillin and 10% were added.
0 μg / ml streptomycin is added and mixed, and the above-mentioned subcultured and collected THP-1 cells are resuspended to a final concentration of 3 × 10 ⁶ cells / ml. This cell resuspension is dispensed into a 24-well cell culture plate in a volume of 0.5 ml. Next, 2.5 μl of DMSO solution in which the pyrrolidinone derivative of the present invention is dissolved in each specific concentration is added to each well. Oxygen the plate 9
Under an atmosphere of a mixed gas of 5% and 5% carbon dioxide, 3
Incubate for 1 hour at 7 ° C. Then, 12-o-tetradecanoylphorbol-13-acetate (hereinafter referred to as "PM
A ”) to a final concentration of 2 μg / ml and polyinosinic acid to a final concentration of 200 μg / ml, respectively, to each well. Further, this plate is cultured at 37 ° C. for 24 hours in an atmosphere of a mixed gas of 95% oxygen and 5% carbon dioxide. After culturing, the supernatant is collected from each well by a Gilson Pipetman, and the amount of IL-1β produced in the supernatant is measured by an enzyme immunoassay kit of Caiman Chemical.

IL-1 without addition of pyrrolidinone derivative
The β production amount is 100, and the concentration of the compound of the present invention that inhibits the IL-1β production amount by 50% is IC ₅₀ , and the results are shown below in μM.

[Table 2]

Example 2 (Measurement of TNFα Release Inhibitory Action) The TNFα release inhibitory action of the pyrrolidinone derivative of the present invention was measured according to the following method. In RPMI 1640 medium (manufactured by Bio Whitker), 10 v / v% fetal bovine serum, 2 mM glutamine, 50 μM 2-mercaptoethanol, 60 μg / ml penicillin and 100 μm.
g / ml streptomycin was added and mixed, and human peripheral blood-derived THP-1 cells (ATCC TIB20
2) is added and suspended. This suspension of THP-1 cells is subcultured at 37 ° C. under an atmosphere of a mixed gas of 95% oxygen and 5% carbon dioxide. A portion of this subcultured suspension is centrifuged at 1200 rpm for 3 minutes at room temperature to recover the subcultured THP-1 cells.

Then in RPMI 1640 medium 2 v / v% fetal bovine serum, 2 mM glutamine, 50 μM 2-mercaptoethanol, 60 μg / ml penicillin and 10%.
0 μg / ml streptomycin is added and mixed, and the above-mentioned subcultured and collected THP-1 cells are resuspended to a final concentration of 3 × 10 ⁶ cells / ml. This cell resuspension is dispensed into a 24-well cell culture plate in a volume of 0.5 ml, and cultured at 37 ° C. for 1 hour.
Next, 2.5 μl of DMSO solution in which the pyrrolidinone derivative of the present invention is dissolved in each specific concentration is added to each well. The plate is incubated at 37 ° C. for 1 hour in an atmosphere of a mixed gas of 95% oxygen and 5% carbon dioxide. Then PMA is added to each well to a final concentration of 2 μg / ml and polyinosinic acid to a final concentration of 200 μg / ml, respectively. Further, this plate is cultured at 37 ° C. for 24 hours in an atmosphere of a mixed gas of 95% oxygen and 5% carbon dioxide. After culturing, the supernatant is collected from each well by a Gilson Pipetman, and the amount of TNFα released in the supernatant is measured using a human TNFα ELISA kit from Zenzyme.

TNFα without addition of pyrrolidinone derivative
The amount of liberation is 100, and the concentration of the compound of the present invention that inhibits the amount of TNFα liberation by 50% is IC _50. The results are shown below in μM.

[Table 3]

Example 3 (Examination of presence or absence of cytotoxicity) The presence or absence of cytotoxicity of the pyrrolidinone derivative was examined according to the following method. In RPMI 1640 medium (manufactured by Bio Whitker), 10 v / v% fetal bovine serum, 2 mM glutamine, 50 μM 2-mercaptoethanol, 60
μg / ml penicillin and 100 μg / ml streptomycin were added and mixed, to which T derived from human peripheral blood was added.
HP-1 cells (ATCC TIB202) are added and suspended. This suspension of THP-1 cells at 37 ° C.
The culture is maintained under an atmosphere of a mixed gas of 95% oxygen and 5% carbon dioxide, and subcultured. A portion of this subcultured suspension is centrifuged at 1200 rpm for 3 minutes at room temperature to recover the subcultured THP-1 cells.

Then, in RPMI 1640 medium, 2 v / v% fetal bovine serum, 2 mM glutamine, 50 μM 2-mercaptoethanol, 60 μg / ml penicillin and 10% were added.
0 μg / ml streptomycin is added and mixed, and the above-mentioned subcultured and collected THP-1 cells are resuspended to a final concentration of 1 × 10 ⁶ cells / ml. This cell resuspension is dispensed into a 24-well cell culture plate in a volume of 1 ml. Next, 5 μl of DMSO solution in which the pyrrolidinone derivative of the present invention is dissolved in each specific concentration is added to each well. The plate was placed in an atmosphere of a mixed gas of 95% oxygen and 5% carbon dioxide at 37 ° C. for 2
Incubate for 4 hours. After the culture, 100 μl of Alamar Blue (manufactured by Biosource) is added to each well, and further cultured at 37 ° C. for 3 hours in an atmosphere of a mixed gas of 95% oxygen and 5% carbon dioxide. After culturing, the supernatant was collected from each well with a Pipetman manufactured by Gilson Company, and the difference in absorbance between the supernatant and 570 nm and 600 nm was measured. 57 without the pyrrolidinone derivative tested
Based on the difference in absorbance between 0 nm and 600 nm, the one in which the difference in absorbance upon addition of the pyrrolidinone derivative of the present invention is significantly reduced is considered to be cytotoxic. According to this test, no cytotoxicity was observed in the pyrrolidinone derivative of the present invention at a concentration of up to 50 μM.

Formulation Example 1 (Tablets) Each tablet was formulated to have the following composition. Compound 9 20 mg Magnesium silicate 20 mg Lactose 98.5 mg Hydroxypropylcellulose 7.5 mg Magnesium stearate 1 mg Hydrogenated vegetable hydrogenated oil 3 mg That is, compound 9, magnesium silicate and lactose were mixed and mixed with hydroxypropylcellulose The mixture was kneaded with the liquid, then granulated to an appropriate particle size, dried and sized, and then magnesium stearate and hydrogenated vegetable hydrogenated oil were added and mixed to obtain uniform granules. Then, a tablet having a diameter of 7.0 mm, a weight of 150 mg and a hardness of 6 kg was prepared with a rotary tableting machine.

Formulation Example 2 (Granule) Compound 9 10 mg Magnesium oxide 40 mg Calcium hydrogen phosphate 38 mg Lactose 10 mg Hydroxypropyl cellulose 20 mg After mixing the respective raw materials except hydroxypropyl cellulose in the above formulation example, hydroxypropyl cellulose was added thereto. The alcohol solution contained was added and kneaded, granulated by an extrusion granulator, and dried to obtain granules. The granules were sized, passed through a 12-mesh screen, and left on the 48-mesh screen to give granules.

Formulation Example 3 (Syrup) Compound 9 1.000 g Sucrose 30.000 g 70 w / v% D-sorbitol 25.000 g Ethyl paraoxybenzoate 0.030 g Propyl paraoxybenzoate 0.015 g Flavor 0.200 g Glycerin 0.150 g 96% Ethanol 0.500 g Purified water Suitable amount Total amount 100 ml Sucrose, D-sorbitol, ethyl paraoxybenzoate,
Propyl paraoxybenzoate and compound 9 are dissolved in 60 g of purified water (warm water). After cooling, a solution of glycerin and 96% ethanol in which the flavor is dissolved is added. Purified water is then added to this mixture to make 100 ml.

Formulation Example 4 (Injection Solution) Hydrochloride of Compound 12 10.0 mg Sodium Chloride 81.0 mg Sodium Bicarbonate 8.40 mg Water for Injection Appropriate amount 10.0 ml Sodium bicarbonate, sodium chloride and hydrochloride of Compound 12 are added to water for injection. It was dissolved and the total volume was adjusted to 10.0 ml to prepare an injection solution.

Formulation Example 5 (Suppository) Compound 9 2 g Macrogol 4000 20 g Glycerin 78 g Compound 9 is added to glycerin and dissolved. Macrogol 4000 is added to this, heated and dissolved, and then poured into a suppository mold and cooled and solidified to produce 1.5 g of each suppository.

[0030]

[Effect] The pyrrolidinone derivative of the present invention has an IL-1β production inhibitory action and a TNFα release inhibitory action,
It is useful as a therapeutic agent for diseases associated with IL-1β and / or TNFα such as rheumatoid arthritis and sepsis.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location A61K 31/44 ACV A61K 31/44 ACV ADZ ADZ // C07D 207/27 C07D 207/27 Z 401 / 06 207 401/06 207 (72) Inventor Shigeo Hatanaka 5-3-1 Tsurugaoka, Oi-cho, Iruma-gun, Saitama Nisshin Flour Milling Co., Ltd. Pharmaceutical Research Laboratory (72) Kenichi Momose Nihombashi Koami, Chuo-ku, Tokyo 19-12 Machi Nisshin Flour Milling Co., Ltd.

Claims (3)

[Claims] 1. A compound of the general formula (I) [Wherein R represents an unsubstituted or substituted phenyl group substituted with a lower alkoxy group or a nitro group, or a pyridyl group] or a pyrrolidinone derivative or a pharmaceutically acceptable salt thereof is contained as an active ingredient. A pharmaceutical composition which is an interleukin-1β production inhibitor and a tumor necrosis factor α release inhibitor. 2. A therapeutic agent for rheumatoid arthritis.
The described pharmaceutical composition. 3. The pharmaceutical composition according to claim 1, which is a therapeutic agent for sepsis.