JPH08165239A - Anti-arteriosclerotic agent - Google Patents

Abstract

PURPOSE: To provide the subject agent useful for preventing or treating arteriosclerosis in order to suppress blood vessel thickening or angiemphraxis often developed after various preventive measures or atheromatous arteriosclerotic lesions involving blood vessel thickening or angiemphraxis as the basic diseases of ischemic organopathy. CONSTITUTION: This anti-arteriosclerotic agent contains, as active ingredient, a pyridinecarboxyimidamide compound of the formula (X is H or NH2 ; Y is 2-chlorophenyl or nitroxyl) or a pharmaceutically permissiblc salt thereof, e.g. 5-amino-N-cyano-N'-[2-(2-chlorophenyl)-ethyl]3-pyridinecarboxyimidamide. This compound of the formula is obtained, for example, by the following process: a cyanopyridine compound is reacted with an alkoxide anion to form an imitate compound, which is then reacted with cyanamide into an N-cyanop pyridinecarboxyimidate, which is then reacted with various amine compounds. This compound of the formula can also be used as an inner membrane thickening inhibitor, inhibitor against reconstriction after surgical operation, lipid metabolism improver or serum lipid depressant.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an antiarteriosclerotic agent, and more particularly to an antiarteriosclerotic agent containing a specific pyridinecarboximidamide compound or a pharmaceutically acceptable salt thereof as an active ingredient. Is.

[0002]

2. Description of the Related Art Myocardial infarction,
Atherosclerosis, which is known as a basic condition of ischemic diseases such as cerebral infarction, renal infarction, and gangrene of the lower limbs, is characterized by remarkable lipid accumulation at the lesion site and intimal thickening mainly composed of vascular smooth muscle cells. , That is, a complex lesion with severe luminal stenosis. It is known that similar lesions (thickening and obstruction of blood vessels due to the proliferation of vascular smooth muscle cells, etc.) often occur after angioplasty, arterial bypass surgery, and organ transplantation, which have been frequently used in recent years. And these are understood as accelerated arteriosclerotic lesions (intimal thickening lesions). Although the mechanism of the development and progression of atherosclerotic lesions or intimal thickening lesions that occur after angioplasty, etc. has not been completely clarified, the common mechanism for both lesions is vascular endothelium. Has been widely accepted as a "damage reaction theory" in which injury to the lung triggers the subsequent abnormal proliferation of vascular smooth muscle cells (Nature, 362).
Vol. 801, 1993). Also, as clinical risk factors for the establishment of intimal hypertrophic lesions formed after atherosclerosis or angioplasty, hypertension, hyperlipidemia, obesity, diabetes, smoking, stress, etc. are mentioned. Among these, hypertension and hyperlipidemia, that is, hypertriglyceridemia and hypercholesterolemia are emphasized. Epidemiological survey on cholesterol (JAMA, 248, 1465, 19
1982) showed that serum cholesterol was 20
Above 0 mg / dl, the incidence of arteriosclerosis-based diseases such as ischemic heart disease and cerebrovascular accident sharply increased, whereas HDL-cholesterol was negatively correlated with the incidence of ischemic heart disease. Indicates. On the other hand, there is controversy as to whether hypertriglyceridemia is an independent risk factor for the development of arteriosclerosis, but at least in postmenopausal women or in the presence of hypercholesterolemia / diabetes, the development of arteriosclerosis occurs. Is known to increase
(Primary Hyperlipidemia Study Group, 1987 Research Report, 1
989; Diabetologia, 32, 300 pages, 1989). More recently, it has become clear that blood triglyceride levels are high in patients with vascular thickening (restenosis) after percutaneous coronary angioplasty (2nd International Symposi
um on Multiple Risk Factors in Cardiovascular Dise
ase, Abstract 72 p. 1992), hypertriglyceridemia is also being recognized as a risk factor for arteriosclerosis. Under these circumstances, agents that reduce risk factors for arteriosclerosis or agents that directly act on arteriosclerotic lesions and improve intimal thickening are considered to be effective in preventing and treating arteriosclerosis. However, despite various studies on drugs that prevent and treat arteriosclerotic lesions including intimal thickening (eg, angiotensin converting enzyme inhibitor cilazapril (restenosis after human percutaneous coronary angioplasty). , Circulation, 85, 1542, 1992), C
a antagonist nicardipine (human coronary atherosclerosis, Circulatio
n, 82, p. 1940, 1990), no effective drug is known yet. An object of the present invention is to provide a drug effective in preventing or treating arteriosclerosis.

[0003]

Means for Solving the Problems As a result of intensive research aimed at achieving the above object, the present inventor has found that a specific pyridine carboxy compound known as a compound having a vasodilatory action and a hypotensive action based on the potassium channel opening action. The present invention was completed as a result of further research conducted by obtaining new knowledge that imidazole compounds have the effects of suppressing intimal thickening and lowering serum triglyceride as described above and suppressing arteriosclerosis. Came to. That is, the drug according to the present invention is an anti-arteriosclerotic agent containing a pyridinecarboxicidamide compound represented by the following general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient, and in another aspect, an inner membrane. It is also a thickening inhibitor, a postoperative (percutaneous coronary angioplasty or bypass) post-restraint inhibitor, a lipid metabolism improving agent, and a serum triglyceride lowering agent.

Embedded image (In the formula, X represents hydrogen or an amino group, and Y represents a 2-chlorophenyl group or a nitroxyl group.)

[Detailed Description of the Invention] Pyridine Carboximidamide Compound The pyridine carboximidamide compound represented by the formula (I) used in the present invention is disclosed in JP-B-6-62567 and WO93-15057. , And is useful as a vasodilator or an antihypertensive agent. Typical preferred specific examples of the compound represented by the formula (I) used for the anti-atherogenic agent of the present invention include the following three (compounds 1 to 3) pyridinecarboximidamide compounds. 1 and 3 are more preferable, and compound 1 is particularly preferable. -Compound 1 5-Amino-N-cyano-N '-[2- (2-chlorophenyl) ethyl] -3-pyridinecarboximidamide-Compound 2 5-Amino-N-cyano-N'-(2-nitroxy Ethyl) -3-pyridinecarboximidamide Compound 3 N-cyano-N ′-[2- (2-chlorophenyl) ethyl] -3-pyridinecarboximidamide The compound represented by the formula (I) as described above is It can be manufactured by any purposeful method, for example, the above-mentioned Japanese Patent Publication No. 6-62567.
It can be produced by the two methods described in Japanese Patent Publication No. WO 93-15057. These methods can be summarized as follows. One method is to react an alkoxide anion on a cyanopyridine compound to produce an imidate compound, which is reacted with cyanamide to convert it into N-cyano-pyridinecarboximidate, which is then reacted with various amine compounds. Is a method of obtaining the desired (N-cyano-N′-substituted) pyridinecarboximidamide compound. Another method is to react the corresponding amide compound with a thiocarbonylating reagent (P ₂ S ₅ , Lawesson's reagent, etc.) to convert it into a thioamide compound, which is then treated with cyanamide in the presence of phosphorus oxychloride and a tertiary amine. It is a method of obtaining the desired (N-cyano-N′-substituted) pyridinecarboximidamide compound by allowing it to act (Patent Publication 6).
No. 62,567, pages 11-14). The compound represented by the general formula (I) used as an active ingredient in the present invention has a basic nitrogen atom and can form an acid addition salt. Examples of the acid to form an acid addition salt include hydrochloric acid,
Inorganic acids such as sulfuric acid, nitric acid, phosphoric acid, or acetic acid, propionic acid, maleic acid, oleic acid, palmitic acid, citric acid, succinic acid, tartaric acid, fumaric acid, glutamic acid,
Examples thereof include organic acids such as pantothenic acid, methanesulfonic acid, toluenesulfonic acid and laurylsulfonic acid. Needless to say, when the acid addition salt is used as a medicine, the acid must be pharmaceutically acceptable.

Antiarteriosclerotic Agent The antiarteriosclerotic agent of the present invention can be administered orally or parenterally (intramuscularly, subcutaneously, intravenously, transdermally) or as a sublingual tablet, suppository and the like. When the compound of the present invention is orally administered as a medicine, it is administered as a tablet, granule, powder or capsule, and when parenterally administered, it is administered as an injection or suspension. To produce these preparations, excipients, binders, disintegrants, lubricants, stabilizers and the like can be added. As an excipient, for example, lactose,
Starch, crystalline cellulose, mannitol, maltose, calcium hydrogen phosphate, light anhydrous silicic acid, calcium carbonate, etc., as a binder, for example, starch, polyvinylpyrrolidone, hydroxypropyl cellulose, ethyl cellulose, carboxymethyl cellulose, gum arabic, etc. As, for example, starch, carboxymethyl cellulose, etc., as a lubricant, for example, magnesium stearate, talc, hardened oil, etc., as a stabilizer,
Examples thereof include lactose, mannitol, maltose, polysorbates, and polyoxyethylene hydrogenated castor oil. Tablets, granules, capsules, using these ingredients,
It can be manufactured into a dosage form such as an injection. When this anti-arteriosclerotic agent is applied to a human, it is preferably administered by intravenous, intramuscular or oral administration. The method of administration and the dose or therapeutically effective amount of the agent of the present invention will vary depending on the circumstances of the individual patient to be treated, for example, weight, sex, sensitivity, administration time, the drug used in combination, the patient or the degree of the disease. Needless to say, the appropriate dose and the number of doses under certain conditions must be determined by a dose determination test by a specialist based on the above guideline, but usually, the dose per adult per day is the active ingredient. about
0.01 to 10 mg / kg (single or divided dose), generally about 0.5 mg, 1 mg, 2 mg, 5 mg on average
Or administration of 10 mg is repeated several times over 2-3 times. The anti-atherosclerotic agent of the present invention is a thickening / stenosis of blood vessels / stenosis / occlusion caused by the proliferation of vascular smooth muscle cells in atherosclerotic lesions, which is a basic lesion of ischemic organ disease, and frequent thickening / stenosis of blood vessels after various operations.・ Prevention of arteriosclerotic lesions to induce reduction of risk factors of occlusion and to directly suppress thickening, stenosis and occlusion of blood vessels caused by atherosclerotic lesions or proliferation of vascular smooth muscle cells after various operations. Alternatively, it can be used as a therapeutic agent. The anti-atherosclerotic agent of the present invention directly affects atherosclerotic lesions with organic stenosis, which is a basic disease of ischemic diseases (angina pectoris, myocardial infarction, etc.) (that is, vascular smooth muscle cell As a drug that suppresses proliferation) or indirectly (ie, through reduction of risk factors such as hyperlipidemia and hypertension), or, for example, various angioplasty procedures (eg, percutaneous coronary angioplasty) Surgery, bypass surgery), transplantation of organs, etc., directly or after vascular obstruction caused by proliferation of vascular smooth muscle cells (eg restenosis after percutaneous coronary angioplasty) As a drug that indirectly suppresses,
Alternatively, a prophylactic and / or therapeutic agent for vascular thickening and blockage (ie, a therapeutic agent for other diseases caused by proliferation of vascular smooth muscle cells)
Is useful as Therefore, the anti-atherosclerotic agent according to the present invention is, in other aspects, an intimal thickening inhibitor, a postoperative (percutaneous coronary angioplasty or bypass angioplasty) inhibitor of restenosis, a lipid metabolism improving agent and serum. It can be said that it is also a sex fat lowering agent.

[0006]

[Examples] The drug efficacy, acute toxicity, and formulation examples of the compound of the present invention are described below. Test Example 1 : Effect on proliferation of cultured vascular smooth muscle cells [Method] Vascular smooth muscle cells were prepared from rat thoracic aorta by the explant method, and Dulbecco's modified Eagle minimal medium containing 10% fetal calf serum (FCS) ( It was cultured in a 96-well plate using DMEM) as a medium. After culturing in DMEM containing 0.5% FCS for 48 hours, the medium was exchanged with DMEM containing 10% FCS, the test drug and 3H-thymidine 0.5 μCi / well were added, and the cells were further cultured for 24 hours. After completion of the culture, the vascular smooth muscle cells were collected with trypsin-EDTA and the radioactivity contained in the cells was measured. As a result, the radioactivity of the test drug was expressed in% when the radioactivity of the vehicle addition group in the presence of 10% FCS was 100% and the radioactivity of the vehicle addition group in the presence of 0.5% FCS was 0%. [Results] Table 1 shows the effects of Compounds 1 to 3 on the proliferation of vascular smooth muscle cells. Table 1 Drug concentration (μM) Proliferation of vascular smooth muscle cells (%) Compound 1 10 98.0 ± 5.5 100 40.1 ± 7.1 ** 1000 -15.0 ± 6.6 ** Compound 2 10 93.6 ± 20.5 100 81.4 ± 16.0 1000 46.1 ± 11.7 * Compound 3 10 49.7 ± 10.7 * 100 8.9 ± 9.2 ** 1000 -20.1 ± 0.6 ** * p <0.05, ** p <0.01 Comparison with vehicle group As shown in Table 1, compounds 1 to 3 are vascular smooth muscle cells. Suppressed the growth of.

Test Example 2 : Inhibitory Effect on Intimal Thickening after Rat Carotid Endothelial Injury by Balloon Catheter [Method] Atherosclerotic lesions and postoperative intimal thickening lesions such as angioplasty are caused by vascular smooth muscle cells. An animal model, which is accompanied by abnormal growth, but well reflects such a pathological condition, that is, a balloon catheter endothelial injury model was used. Powell's method (J. Cardiovasc.Pharmacol., 16
Vol., P. S42, 1990). 1
1-week-old male Wistar rats were used. The neck of the rat was incised under anesthesia with sodium pentobarbital, and a balloon catheter (Fogherty, 2F) was inserted from the left external carotid artery to the origin of the common carotid artery. After inflating the balloon with physiological saline to the extent that the artery was lightly dilated, the endothelium was injured by pulling the catheter to the external carotid artery. After repeating this operation three times, the catheter was removed and the external carotid artery was ligated. 14 days later, thoracotomy was performed under anesthesia with diethyl ether, and heparin was added from the left ventricle (10 U / m
l) After perfusion with physiological saline and then with 4% paraformaldehyde solution, the left common carotid artery was isolated. The excised artery was fixed in a buffered paraformaldehyde solution for a further 48 hours or more. The fixed carotid artery was stained with hematoxylin and eosin and then imaged under a microscope (Ze
iss) to measure the area of the media and intima,
The media area ratio was calculated. The numerical values of the results are shown as the average value ± standard error. The test drug was orally administered daily from 6 days before the surgery to 14 days after the surgery, or an osmotic pump containing the drug (A
lza) was continuously administered by implanting it intraperitoneally 2 days before the operation. The intimal thickening inhibitory activity of the drug was determined by the following formula. Intimal thickening inhibition rate (%) = (1-T / C) x 100 T: Intima / media ratio of drug-administered rat blood vessels C: Intimal / media ratio of vehicle rat blood vessels [Results] It is as shown in Table 2. Table 2 Drug dose Number of intima / media area ratio (inhibition rate,%) Vehicle 11 1.25 ± 0.11 Compound 12 mg / kg / day, po 8 0.96 ± 0.10 (23.2) Vehicle 11 1.26 ± 0.14 Compound 1 5 mg / kg / day, po 13 0.72 ± 0.06 ** (43.0) Medium 13 1.08 ± 0.08 Compound 1 1 mg / kg / day, ip 14 1.06 ± 0.09 (2.0) 3 mg / kg / day, ip 10 0.68 ± 0.10 ** (37.0) Medium 14 1.25 ± 0.08 Enalapril 30 mg / kg / day, ip 4 0.44 ± 0.21 ** (64.8) Medium 12 1.10 ± 0.08 Verapamil 45 mg / kg / day, ip 9 1.33 ± 0.20 (-20.7) Hydralazine 2 mg / kg / day, ip 15 1.08 ± 0.07 (2.0 ) Vehicle 12 1.12 ± 0.05 Nifedipine 3 mg / kg / day, ip 10 1.00 ± 0.10 (11.0) Cromakalim 3 mg / kg / day, ip 12 0.74 ± 0.06 ** (34.3) ** p <0.01 Comparison with vehicle group As is clear from Table 1, Compound 1 was found to increase body weight after oral administration of 5 mg / kg / day and continuous intraperitoneal administration of 3 mg / kg / day. The intimal thickening induced by endothelial injury was significantly suppressed without any effect. At this time, the positive control enalapril (angiotensin converting enzyme inhibitor) also suppressed the intimal thickening. On the other hand, Ca antagonists (verapamil, nifedipine) and hydralazine, which are known as antihypertensive agents, did not suppress the intimal thickening at doses showing the same degree of hypotension as Compound 1.

Test Example 3 : Action on lipid metabolism in normal rats [Method] SD type (5-week-old) rats were orally administered with the test drug for 10 consecutive days, and blood was collected 4 hours after the final administration. After blood was centrifuged to obtain serum, lipid parameters in the serum were measured by an enzymatic method. [Results] Table 3 shows serum lipid levels in normal rats. Table 3 Compound 1 Actions on lipid metabolism in normal rats Drug dose Example Number Total cholesterol Total triglyceride (mg / dl) (mg / dl) Vehicle 12 69.2 ± 1.9 91.0 ± 6.8 Clofib 160 mg / kg 11 58.7 ± 3.7 ** 61.3 ± 6.8 ** ^rate (84.8) (67.4) Compound 13 mg / kg 13 75.8 ± 1.9 53.1 ± 5.0 ** (109.5) (58.4) The value in parentheses indicates the ratio (%) to the value of the medium group. * p <0.05, ** p <0.01 Comparison with vehicle group Compound 1 showed a tendency to decrease serum total cholesterol without affecting body weight gain in normal rats, and significantly decreased serum total triglyceride in a dose-dependent manner. I'm sorry. In addition, Compound 1 significantly increased HDL cholesterol (vehicle group 30.4 mg / dl vs. Compound 1 37.4 mg / dl),
As a result, the arteriosclerosis index was decreased as compared with the control group (Compound 1 was 1.0 while vehicle group was 1.3).

Test Example 4 Effect on Lipid Metabolism of Rats Loaded with High Fat Diet Method: SD (5-week-old) rats were fed a high fat diet containing 0.75% cholesterol and 7.5% lard for 10 days. The test drug was orally administered every day for 10 days from the start of high fat diet loading, and blood was collected 4 hours after the final administration. After blood was centrifuged to obtain serum, lipid parameters in the serum were measured by an enzymatic method. [Results] Table 4 shows serum lipid levels in rats fed a high fat diet. Table 4 Effects of Compound 1 on lipid metabolism in rats fed a high fat diet Drug doses Total cholesterol Total triglyceride (mg / dl) (mg / dl) Vehicle 8 242.2 ± 19.7 183.6 ± 23.4 Clofib 160 mg / kg 8 158.6 ± 17.0 ** 97.2 ± 24.2 ** ^Latte (65.5) (52.9) Compound 1 1 mg / kg 7 224.3 ± 18.4 105.5 ± 15.4 * (92.6) (57.4) 3 mg / kg 8 223.3 ± 13.7 82.1 ± 7.7 ** (92.2 ) (44.7) 10 mg / kg 8 220.9 ± 11.1 70.6 ± 6.2 ** (91.2) (38.4) The value in parentheses indicates the ratio (%) to the value of the medium group. * p <0.05, ** p <0.01 Comparison with vehicle group Compound 1 showed a tendency to decrease serum total cholesterol without affecting food intake and body weight gain in high fat diet-loaded rats, and dosed serum total triglyceride It was significantly decreased in a dependent manner.

Test Example 5 : Effect on the lipid metabolism of genetically hyperlipidemic rats [Method] Zucker rat which is a model animal for type IV hyperlipidemia
(14 weeks old), the test drug was orally administered every day for 14 days, and blood was collected 4 hours after the final administration. After blood was centrifuged to obtain serum, lipid parameters in the serum were measured by an enzymatic method. [Results] Table 5 shows serum lipid levels in Zucker rats. Table 5 Effects of Compound 1 on lipid metabolism in genetically hyperlipidemic rats Drug dose Number of samples Total cholesterol Total triglyceride (mg / dl) (mg / dl) Vehicle 5 143.9 ± 32.3 845.0 ± 265.4 Compound 1 1 mg / kg 6 151.6 ± 3.4 695.8 ± 184.3 10 mg / kg 4 142.2 ± 5.2 305.3 ± 24.8 * * p <0.05, Comparison with vehicle group Compound 1 dose-dependently decreased serum total triglyceride without affecting body weight gain Let

Test Example 6 : Acute Toxicity [Method] 50, 25 in 5 week-old SD male rats of 3 groups per group.
A single oral dose of 0, 500 mg / kg of Compound 1 was administered. [Results] All animals died in the 250 and 500 mg / kg groups, whereas no deaths were observed in the 50 mg / kg group. In general observation after administration, redness of limbs and auricles was observed at all doses, which is considered to be due to the vasodilatory action which is the pharmacological action of Compound 1. No abnormalities were observed by gross necropsy in both dead and surviving cases. Also, in this test
LD50 by Probit method could not be calculated, but LD50
Is considered to be in the range of 50-250 mg / kg.

The formulation examples of the present invention are shown below. Formulation example 1 Compound of the present invention 2.0 mg Lactose 80.2 mg Crystalline cellulose 31.2 mg CMC sodium 6.0 mg Magnesium stearate 0.6 mg Total 120.0 mg The above ingredients are mixed and pressed to give tablets.

Formulation Example 2 Compound of the present invention 2.0 mg Lactose 81.8 mg Corn starch 55.85 mg Magnesium stearate 0.35 mg Total 140.0 mg The above ingredients are mixed and filled into capsules to give capsules.

[0014]

As described above, the anti-arteriosclerotic agent according to the present invention is effective for thickening blood vessels, which often occurs after various operations.
It is useful as a prophylactic or therapeutic agent for arteriosclerosis for suppressing atherosclerotic lesions accompanied by thickening / occlusion, which is a basic disease of obstruction or ischemic organ disease.

Claims (6)

[Claims] 1. An anti-atherosclerotic agent containing a pyridinecarboximidamide compound represented by the following general formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient. Embedded image (In the formula, X represents hydrogen or an amino group, and Y represents a 2-chlorophenyl group or a nitroxyl group.) 2. X is an amino group or hydrogen bonded to the 5-position of the pyridine ring, and Y is a 2-chlorophenyl group.
The anti-arteriosclerotic agent according to claim 1. 3. An intimal thickening inhibitor comprising the pyridinecarboximidamide compound represented by the general formula (I) according to claim 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient. . 4. A percutaneous coronary angioplasty containing the pyridinecarboximidamide compound represented by the general formula (I) according to claim 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient. Inhibitor for restenosis after surgery or bypass surgery. 5. A lipid metabolism-improving agent containing the pyridinecarboximidamide compound represented by the general formula (I) according to claim 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient. 6. A serum triglyceride-lowering composition containing the pyridinecarboximidamide compound represented by the general formula (I) according to claim 1 or 2 or a pharmaceutically acceptable salt thereof as an active ingredient. Agent.