JPH093019A - Amide derivative and pharmaceutical preparation containing the same - Google Patents

Abstract

PURPOSE: To obtain a new amide derivative having proliferation-inhibitory effect on several mesodermic cells such as smooth muscle cells, nephromesangial cells and fibroblasts, thus useful for treating the reconstriction after percutaneous coronary dilative operation, and inflammatory and detail proliferative fibrosclerosis such as chronic glomerulonephritis. CONSTITUTION: This new compound is expressed by formula I [R1 -R3 are each H, an alkyl, an alkoxyl or an aryl(oxy); Ar is an aryl; (n) is 0 or 1], e.g. 2-(2,5- dimethoxycinnamoylamino)thiazole. The compound of formula I is obtained by reaction of a compound of formula II with a carboxylic acid activator such as thionyl chloride, phosphorus pentachloride or carbodiimide to form a carboxyl-reactive derivative, which is then reacted with a compound of the formula Ar-NH2 .

Description

Detailed Description of the Invention

[0001]

INDUSTRIAL APPLICABILITY The present invention has an inhibitory effect on the proliferation of several mesodermal cells such as smooth muscle cells, renal mesangial cells, fibroblasts, restenosis after PTCA, and chronic glomerulonephritis. The present invention relates to an amide derivative capable of effectively treating inflammatory and cell proliferative fibrosclerosis, and a pharmaceutical preparation containing the same.

[0002]

2. Description of the Related Art It is known that the onset of pathological conditions in angina, myocardial infarction and the like is largely caused by coronary arteriosclerosis which precedes it. The narrowing of the lumen and the loss of elasticity of blood vessels caused by arteriosclerosis lead to nutritional and oxygen deficiency in myocardial tissue and induce the above-mentioned pathological conditions. The narrowing of the vascular lumen is caused by the accumulation of foamed macrophages and cholesterol in the inner wall, migration of vascular medial smooth muscle cells into the intima, and cell fibrous intimal thickening caused by proliferation in the intima. It is said to be the cause. Recently, percutaneous coronary dilation (Percutaneous Transluminal Corona Dilatation) has been used as a method for surgically treating arteriosclerotic blood vessels with stenosis.
ry Angioplasty (PTCA) is becoming popular. In the PTCA operation, a balloon catheter is remotely inserted from the femoral artery or the like, the balloon is inflated at the stenosis, and the blood vessel is physically expanded.

However, the biggest problem with this treatment method is that stenosis occurs again in 30 to 50% of the treated cases within 3 to 6 months after the treatment (Spencer B. King; Am. J. Candiol, 198
7,60 (3), 1B). No cholesterol deposition was observed in this restenosis, but rather most of it was composed of smooth muscle cells and the intercellular matrix produced by these cells.
So-called cell fibrous intimal thickening. Therefore, PTCA
As a method for preventing post-restenosis and eventually treating arteriosclerosis, it is effective to suppress migration and proliferation of smooth muscle cells generated in the lumen of blood vessels. At present, as such conventional techniques, JP-A-6-135829 and JP-A-6-305966 have been reported, but the appearance of an active substance that more strongly suppresses the proliferation of smooth muscle cells has been reported. Strongly desired.

Similarly, in chronic nephritis, it is known that proliferation of mesangium cells and proliferation of intercellular matrix in the glomerulus cause renal sclerosis and cause renal function decline. In addition, in liver fibrosis, stellate cells of mesenchymal cells and abnormal production of collagen are pathological conditions such as lung fibrosis, abnormal proliferation of fibroblasts and inflammation of intercellular matrix that occur after inflammation in peritoneal thickening that occurs during peritoneal dialysis. It is said to be the cause of. Therefore, there is a strong demand for the development of a drug that can effectively treat the abnormal proliferation of these cells and the proliferation of intercellular matrix.

[0005]

Therefore, the present invention is based on the PT
Provided are a compound which is useful as an agent for preventing restenosis after CA surgery, an agent for preventing restenosis after transplantation of an autologous blood vessel and an artificial blood vessel, and a therapeutic agent and a preventive agent for arteriosclerosis, and an agent for preventing vascular wall thickening containing the same The purpose is to In addition, it is also provided as a compound effective for treating fibrosis or fibrous sclerosis caused by inflammation-induced cell proliferation and intercellular matrix hyperplasia.

[0006]

Means for Solving the Problems As a result of diligent investigations on the pharmacological activity of the novel amide derivatives, the present inventors surprisingly found that the amide compound of the present invention was PDGF.
The present invention has been completed by finding out that the proliferation effect of cultured smooth muscle cells induced by erythrocyte is specifically suppressed. The present invention is as follows.

An amide derivative represented by the following general formula (1).

[0008]

Embedded image

(In the formula 1, R ₁ , R ₂ and R ₃ are the same or different and each represents hydrogen, an alkyl group, an alkoxy group, an aryl group or an aryloxy group, and Ar is an aryl group or n.
Represents an integer of 0 or 1. )

The present invention is also a pharmaceutical preparation containing the above amide derivative. Further, the present invention is a vascular wall thickening preventive agent comprising the above amide derivative.

As used herein, the term "alkyl" means a straight or branched chain group, which includes a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group and a sec- group.
Examples thereof include, but are not limited to, a butyl group, an isobutyl group, a tert-butyl group, a pentyl group, an isopentyl group, a hexyl group, a heptyl group, an octyl group, a nonyl group, and a decyl group.

As used herein, the term "alkoxy" means-
OR ₄ (R ₄ is an alkyl group) means a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a butoxy group, a sec-butoxy group, an isobutoxy group, a tert-butoxy group and the like. However, the present invention is not limited to these.

As used herein, the term "aryl" means a substituted or unsubstituted carbocyclic or heterocyclic aromatic group (substituents are halogeno group, nitro group, cyano group, alkyl group,
An alkoxy group and a halogen-substituted alkyl group), which includes a phenyl group, 1- or 2-
It includes, but is not limited to, naphthyl group, 2-, 3- or 4-pyridyl group, 2- or 3-furyl group, 2-4- or 5-thiazolyl group and the like.

In the present specification, "aryloxy" means -OR ₅ (R ₅ is an aryl group), and includes phenoxy group, 1-naphthoxy group, 2-naphthoxy group and the like. It is not limited to.

The term "halogen" as used herein means a group derived from a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

The term "halogen-substituted alkyl" as used herein refers to the above alkyl group substituted with one or more halogens, including chloromethyl group, trifluoromethyl group, 2,2-difluoroethyl group. Groups and the like, but are not limited thereto.

The compounds of the present invention are all novel compounds which have not been published in the literature, and the compound represented by the general formula (1) is, for example, a carboxylic acid derivative represented by the following general formula (2):
It can be produced by reacting a carboxylic acid activator to give a reactive derivative at the carboxyl group, and then reacting it with an amine derivative represented by the following general formula (3).

[0018]

Embedded image

(In the formula 2, R ₁ , R ₂ , R ₃ and n have the same meaning as in the general formula (1).)

[0020]

Embedded image

(In the formula 3, Ar has the same meaning as in the general formula (1).)

In the reaction between the carboxylic acid derivative (2) and the carboxylic acid activator, the carboxylic acid activator is
For example, thionyl chloride, phosphorus pentachloride, chloroformate (methyl chloroformate, ethyl chloroformate), oxalyl chloride, carbodiimides (eg, N, N'-dicyclohexylcarbodiimide (DCC), 1-ethyl-3-
(3-Dimethylaminopropyl) carbodiimide (WS
C)) etc., but carbodiimides and N-hydroxybenzotriazole, 4-dimethylaminopyridine or hydroxysuccinimide may be used in combination. This reaction is usually carried out by halogenated hydrocarbons such as methylene chloride and chloroform, ethers such as tetrahydrofuran (THF), dioxane, dimethyl ether, diethyl ether and isopropyl ether,
It is carried out in the presence of N, N-dimethylformamide, N, N-dimethylacetamide or a mixed solvent thereof. The reaction temperature is usually -10 ° C to 50 ° C.

In this reaction, when thionyl chloride, oxalyl chloride or phosphorus pentachloride is used as the carboxylic acid activator, an acid halide is obtained as a reactive derivative, and chloroformic acid ester is used as the carboxylic acid activator. In that case, a mixed acid anhydride as a reactive derivative is obtained, and when a carbodiimide is used as a carboxylic acid activator, an active ester is obtained as a reactive derivative.

When the reactive derivative at the carboxyl group of the carboxylic acid derivative (2) is reacted with the amine derivative (3), when the reactive derivative is an acid halide, for example, in a solvent such as methylene chloride, tetrahydrofuran or acetone, It is carried out under anhydrous or hydrous conditions in the presence of a deoxidizing agent (pyridine, triethylamine, potassium carbonate, potassium hydrogen carbonate, sodium hydrogen carbonate, etc.). The reaction temperature is -50 ° C to 100 ° C, preferably -10 ° C to 30 ° C. When the reactive derivative is an active ester or a mixed acid anhydride, it can be carried out in a solvent similar to the solvent used in the reaction of the carboxylic acid derivative (3) with the carboxylic acid activator. In this case, the reaction temperature is usually 0 to 30 ° C., and the reaction time is usually 1 to 5 hours. The amide derivative (1) thus produced can be isolated and collected by a separation / purification means known per se (eg, chromatography, recrystallization) and the like.

The amide derivative of the present invention may optionally be a pharmaceutically acceptable salt thereof, for example, an alkali metal salt such as sodium salt or potassium salt, an alkaline earth metal salt such as calcium salt or magnesium salt. , Ammonium salts, salts with organic bases such as triethylamine salt, pyridine salt, tromethamine salt, dicyclohexylamine salt, formate salt, toluenesulfonate salt, trifluoroacetate salt, hydrochloride salt, sulfate salt, nitrate salt and the like, arginine,
Examples thereof include amino acid salts such as lysine and aspartic acid.

The amide derivative of the present invention can be administered orally or parenterally (for example, intravenously, intramuscularly, subcutaneously) as a vascular wall thickening preventive agent. The dose of the active ingredient compound of the present invention will vary depending on the age, body weight and symptoms of the patient, but is usually about 0.1 to 1000 mg / kg per day,
Preferably, 1-100 mg / kg is administered in 1 to 3 divided doses.

The compound of the present invention is used as an active ingredient or one of the active ingredients, alone or in combination with a pharmaceutical carrier, according to known formulation techniques, such as tablets, powders, capsules, granules, syrups, solutions, suspensions, injections, Any formulation suitable for administration of eye drops or suppositories can be adopted. Specific formulation carriers include starch, sucrose, lactose, methyl cellulose, carboxymethyl cellulose, crystalline cellulose, sodium alginate, calcium hydrogen phosphate, magnesium aluminometasilicate, anhydrous silicic acid,
And excipients such as synthetic aluminum silicate, binders such as hydroxypropylcellulose, hydroxypropylmethylcellulose, gelatin and polyvinylpyrrolidone, disintegrants such as carboxymethylcellulose calcium, crosslinked sodium carboxymethylcellulose and crosslinked polyvinylpyrrolidone, magnesium stearate and Lubricants such as talc, cellulose acetate phthalate, hydroxypropylmethyl cellulose acetate succinate, coating agents such as methacrylic acid and methyl methacrylate methyl copolymer, dissolution aids such as polyethylene glycol, sodium lauryl sulfate, lecithin, sorbitan monoole. Ate, polyoxyethylene cetyl ether, sucrose fatty acid ester, polyoxyethylene hydrogenated castor oil and guar Emulsifiers monostearate, etc., chelating agents such as EDTA, buffering agents, humectants, preservatives, it may be mentioned groups such as cocoa butter and Uitebuzoru W35.

[0028]

The present invention will be described in more detail with reference to examples and test examples, but the present invention should not be limited to these examples and test examples.

Example 1 2- (2,5-dimethoxycinnamoylamino) thiazo
Synthesis of 2-aminothiazole (1.20 g, 12 mmol) in N, N
To a solution of dimethylformamide (20 ml) 2,5-dimethoxycinnamic acid (2.08 g, 10 mmol) is added.
Further, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1.91 g, 10 mmol) and dimethylaminopyridine (122 mg, 1 mmol) were added,
The mixture was stirred at room temperature for 24 hours. Water was added to the reaction solution and extracted with ethyl acetate. The ethyl acetate layer was washed with water and saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue obtained was subjected to silica gel chromatography, and the elution fraction of 1% methanol-chloroform was used. The structure of the compound is represented by the following formula (4), and the target compound (1.
34 g, 46%) was obtained.

Mp: 220.0-221.0 ° C. ¹ H-NMR (400 MHz, DMSO ₄ ) δ (ppm):
3.79 (3H, s), 3.85 (3H, s), 7.00
(1H, d, J = 15.93Hz), 7.01 (1H, d,
J = 2.75), 7.02 (1H, s), 7.11 (1
H, d, J = 2.75Hz), 7.20 (1H, d, J =
3.66Hz), 7.49 (1H, d, J = 3.66Hz),
7.88 (1H, d, J = 15.93Hz) MS (FAB): 292 (M + 1)

[0031]

Embedded image

Example 2 Synthesis of 4'-cyano-4-phenylbenzanilide 4-biphenylcarbochloride (1.08 g, 5 mmo)
Dimethylaminopyridine (733 mg, 6 mmol) and 4-cyanoaniline (591 mg, 5 mmol) were added to a solution of l) in methylene chloride (30 ml), and the mixture was stirred at room temperature for 12 hours. The reaction solution was diluted with methylene chloride, washed with 1N aqueous hydrochloric acid solution, saturated aqueous sodium hydrogen carbonate solution and saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. A colorless solid (recrystallized: ethyl acetate-hexane) having the following properties is shown in the formula (5) below from the elution fraction of 1% methanol-chloroform.
The target compound (990 mg, 66%) was obtained.

Mp: 215.0-216.0 ° C. ¹ H-NMR (400 MHz, CDCl ₃ ) δ (ppm):
7.41 (1H, t, J = 7.80Hz), 7.45 (2
H, t, J = 7.80 Hz), 7.62 (2H, d, J =
7.80Hz), 7.64 (2H, d, J = 8.40Hz),
7.72 (2H, d, J = 8.10Hz), 7.81 (2
H, d, J = 8.40 Hz), 7.93 (2H, d, J =
8.10 Hz), 8.03 (1H, bs) MS (FAB): 299 (M + 1)

[0034]

[Chemical 6]

(Example 3) 2 ', 5'-dimethoxy-4-phenylbenzanilide
According to the method of Synthesis Example 1 above, a target compound having the structure shown in the following formula (6) and having the following properties was produced.

Mp: 139.0-140.0 ° C. ¹ H-NMR (400 MHz, CDCl ₃ ) δ (ppm):
3.82 (3H, s), 3.90 (3H, s), 6.61
(1H, d, J = 8.80Hz), 6.84 (1H, d, J
= 8.80Hz), 7.40 (1H, t, J = 7.30H
z), 7.47 (2H, J = 7.30Hz), 7.63 (2
H, d, J = 7.30 Hz), 7.72 (2H, d, J =
8.60Hz), 7.97 (2H, d, J = 8.60Hz),
8.31 (1H, s), 8.62 (1H, bs) MS (FAB): 334 (M + 1)

[0037]

Embedded image

(Example 4) 2- (2,5-dimethoxycinnamoylamino) -6-
Synthesis of Methyl Pyridine According to the method of Example 2, a target compound having the structure shown below in the formula (7) and having the following properties was produced.

¹ H-NMR (400 MHz, CDCl ₃ )
δ (ppm): 2.46 (3H, s), 3.78 (3H,
s), 3.82 (3H, s), 6.68 (1H, d, J =
15.6 Hz), 6.83-6.91 (3H, m), 7.00
(1H, s), 7.62 (1H, dd, J = 8.4, 7.
6 Hz), 7.95 (1 HdJ = 15.6 Hz), 8.1
7 (1H, d, J = 8.4Hz), 8.53 (1H, bs) MA (FAB): 299 (M + 1)

[0040]

Embedded image

(Example 5) 2- (2,4-dimethoxycinnamoylamino) -6-
Synthesis of Methyl Pyridine According to the method of Example 2, a target compound having the structure shown below in the formula (8) and having the following properties was produced.

Mp: 138-143 ° C. ¹ H-NMR (400 MHz, CDCl ₃ ) δ (ppm):
2.47 (3H, s), 3.84 (3H, s), 3.86
(3H, s), 6.46 (1H, s), 6.51 (1H,
d, J = 8.4Hz), 6.61 (1H, d, J = 15.8H)
z), 6.89 (1H, d, J = 7.6Hz), 7.41 (1
H, d, J = 8.4 Hz), 7.61 (1H, dd, J =
8.0, 7.6 Hz), 7.90 (1H, d, J = 15.8H
z), 8.16 (1H, d, J = 8.0Hz), 8.19 (1
H, s) MS (FAB): 299 (M + 1)

[0043]

Embedded image

(Example 6) 2- (3,5-dimethoxycinnamoylamino) -6-
Synthesis of Methyl Pyridine According to the method of Example 1, a target compound having the structure shown in the following formula (9) and having the following properties was produced.

Mp: 136-138 ° C. ¹ H-NMR (400 MHz, CDCl ₃ ) δ (ppm):
2.46 (3H, s), 3.79 (3H, s), 6.48
(1H, d, J = 15.4Hz), 6.48 (1H, s),
6.63 (1H, s), 6.92 (1H, d, J = 7.6H
z), 7.63 (1H, dd, J = 8.0, 7.6Hz),
7.67 (1H, d, J = 15.4Hz), 8.16 (1
H, d, J = 8.0 Hz), 8.69 (1 H, s) MS (FAB): 299 (M + 1)

[0046]

Embedded image

Example 7 Combination of 2- (4-biphenylcarbonylamino) pyridine
According to the method of Synthesis Example 1, a target compound having the structure shown in the following formula (10) and having the following properties was produced.

Mp: 164-165 ° C. ¹ H-NMR (400 MHz, CDCl ₃ ) δ (ppm):
7.02-7.06 (1H, m), 7.37-7.41 (1
H, m), 7.44-7.49 (2H, m), 7.61-
7.63 (2H, m), 7.70 (2H, d, J = 8.8
0Hz), 7.73-7.77 (1H, m), 8.00 (2
H, d, J = 8.80 Hz), 8.21-8.23 (1H,
m), 8.42 (1H, d, J = 8.40Hz), 9.02
(1H, s) MS (FAB): 275 (M + 1)

[0049]

Embedded image

(Example 8) 2- (4-biphenylcarbonylamino) -4,6-di
Synthesis of Methyl Pyridine According to the method of Example 1, a target compound having the structure shown in the following formula (11) and having the following properties was produced.

Mp: 156-159 ° C. ¹ H-NMR (400 MHz, CDCl ₃ ) δ (ppm):
2.38 (3H, s), 2.43 (3H, s), 6.78
(1H, s), 7.40 (1H, m), 7.47 (2H,
m), 7.63 (2H, m), 7.71 (2H, d, J =
8.6Hz), 8.00 (2H, d, J = 8.6Hz), 8.
07 (1H, s), 8.60 (1H, s) MS (FAB): 303 (M + 1)

[0052]

[Chemical 12]

Example 9 2- (5- (3,4,5- (trimethoxy) phenyl)-
Penta-2,4-dienoyl) amino-6-methylpyri
Synthesis of Zin According to the method of Example 1, a target compound having the structure shown in the following formula (12) and the following properties was produced.

Mp: 169-171 ° C. ¹ H-NMR (400 MHz, CDCl ₃ ) δ (ppm):
2.46 (3H, s), 3.88 (6H, s), 3.90
(3H, s), 6.07-8.15 (9H, m), 8.2
7 (1H, bs) MS (FAB): 377 (M + 1)

[0055]

Embedded image

Test Example Proliferation Inhibitory Action of Cultured Smooth Muscle Cells The median smooth muscle layer was taken out from the thoracic aorta of a 6-week-old Wistar male rat (manufactured by Charles River Japan) and 1 mm ²
After being cut into pieces, they were attached to a 25 cm ³ culture flask (made by Corning) and Dulbecco modifi containing 10% serum was added.
The cells were cultured in an ed eagle medium (hereinafter abbreviated as DMEM: manufactured by Nissui Co., Ltd.) in an incubator under conditions of 37 ° C. and 95% O ₂ + 5% CO ₂ for _{2 to} 3 weeks. Elongated and dividing cells from the section were collected as primary culture smooth muscle cells. The primary cultured smooth muscle cells were DME containing 10% fetal bovine serum (manufactured by Gibco) in a Petri dish (manufactured by Corning) with a diameter of 9 cm.
Cultured in M and confluent 3-4 days 3
It was subcultured in double dose. The test was performed using cells during which this operation was repeated 4 to 8 times, that is, between passage numbers 5 and 9. The cultured smooth muscle cells were plated in a 24-well plate (Falcon) with 8 × 10 ⁴ smooth muscle cells / well / 700 μl D
Seeding was done at the ratio of MEM. After overnight, it was made serum-free and cultured in an incubator for 2 days. Under this condition, the cultured smooth muscle cells are in the G ₀ phase (rest phase) and do not divide.

The hydroxamic acid derivative used in the test was dim
After dissolving in ethylsulfoxide (DMSO), 4% bovine se
First, it was diluted 100 times with DMEM containing rum albumin, and further 20 times with DMEM. That is 200
A 0-fold diluted test solution was added to cells under the above conditions together with a growth stimulating factor. The growth factors used were 10% fetal bovine serum, 10 ng / ml platelet growth factor (PDGF), 5 ng / ml fibroblast growth factor (FGF), 5 ng / ml epidermal growth factor (EGF), 20 ng / ml insulin-like. Growth factor-1
(IGF-1) was used respectively. [3H] -methyl thymidine (manufactured by Amersham) was added 18 hours after stimulation at a rate of 0.5 μCi / ml / well, and the medium was removed 6 hours later.
The cells contained 1 ml of phosphate buffer (Ca ²⁺ , Mg ²⁺ -fre
After washing twice with e), elution was carried out with 500 μl of Tris-HCl buffer containing 0.1% SDS. After thorough stirring, 100 μl of 500 μl was soaked in filter paper and air dried. The filter paper was washed with a 5% trichloroacetic acid (containing 100 mM sodium pyrophosphate) solution 3 times for 15 minutes each and twice for 15 minutes each with ethanol at 4 ° C., and then air-dried.
Submerged at the bottom of the vial containing ml. This was measured with a liquid scintillation counter (made by Packard) for 5 minutes. Table 1 shows the 50% smooth muscle cell growth inhibitory activity concentration (IC ₅₀ ) of the drug stimulated with 10% fetal bovine serum and 10 ng / ml platelet growth factor (PDGF).

[0058]

[Table 1]

(Acute toxicity) An acute toxicity test was carried out by oral and intravenous administration using male ICR mice (5 weeks old), and as a result, the LD ₅₀ value of the amide derivative of the present invention was 320 mg / kg or more. Yes, higher safety was confirmed compared to efficacy.

[0060]

INDUSTRIAL APPLICABILITY The novel amide derivative and the pharmaceutical preparation containing the same according to the present invention have an inhibitory effect on the proliferation of several mesodermal cells such as renal mesangium cells and fibroblasts typified by smooth muscle cells. Therefore, it is useful as a drug capable of effectively treating inflammatory and cell proliferative fibrosclerosis represented by restenosis after PTCA, chronic glomerulonephritis and the like.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical indication C07C 231/02 9547-4H C07C 231/02 233/02 9547-4H 233/02 233/64 9547- 4H 233/64 233/88 9547-4H 233/88 255/60 9357-4H 255/60 C07D 213/75 C07D 213/75 277/46 277/46

Claims (2)

[Claims] 1. An amide derivative represented by the following general formula (1). Embedded image (In the formula 1, R ₁ , R ₂ and R ₃ are the same or different and each represents hydrogen, an alkyl group, an alkoxy group, an aryl group or an aryloxy group, Ar is an aryl group, n is 0 or 1
Indicates an integer. ) 2. A pharmaceutical preparation comprising the amide derivative according to claim 1.