JP3074043B2 - Atherosclerosis treatment / prevention agent - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an agent for treating and preventing arteriosclerosis.

[0002]

2. Description of the Related Art Atherosclerosis is closely related to stroke, myocardial infarction, hypertension, etc., which are the leading causes of death in Japanese people, and great efforts have been made to develop therapeutic and preventive drugs therefor. Arteriosclerosis is a general term for lesions showing localized thickening and hardening of the arterial wall, and is used in the present application in the broadest meaning including atherosclerosis, medial sclerosis, and arteriosclerosis.

[0003] Conventionally, anti-lipidemic drugs which lower blood lipids, particularly cholesterol, have been mainly used for the treatment and prevention of arteriosclerosis, but they are not effective enough.

[0004]

At present, there is no known drug effective for treating or preventing arteriosclerosis, and a promising new drug is eagerly needed.

[0005]

There are various theories regarding the etiology of arteriosclerosis. Atherosclerosis, which forms the core of atherosclerotic lesions, is caused by low density lipoprotein (hereinafter abbreviated as LDL) in plasma. ) Especially acetyl LDL or oxidized L
Penetration of so-called denatured LDL such as DL into the artery wall;
It is almost theorized that macrophage and their uptake by smooth muscle cells migrating to the intima are factors (Ayumi of Medicine 147, No. 9).
761 1988).

[0006] As the etiology of arteriosclerosis has been elucidated in this way, a new method of developing a therapeutic / prophylactic agent for arteriosclerosis has been found. That is, various compounds are added to LDL, and 2,2′-azobis (2-amidinopropane) dihydrochloride (hereinafter abbreviated as AAPH), which is a water-soluble radical initiator, or 2, which is a lipophilic radical initiator, 2
Oxidation with '-azobis (2,4-dimethylvaleronitrile) (hereinafter abbreviated as AMVN) to produce peroxide of phosphatidylcholine (hereinafter abbreviated as PC) and cholesterol ester (hereinafter abbreviated as CE) (hereinafter abbreviated as CE) Each PC-OO
H, abbreviated as CE-OOH), and the L of each compound was measured.
A new arteriosclerotic drug can be developed by examining the effects during DL. The present inventors have conducted intensive studies using this method, and have found that a compound having the following general formula can be an excellent therapeutic / prophylactic agent for arteriosclerosis, and completed the present invention. Therefore, an object of the present invention is to provide an excellent agent for treating and preventing arteriosclerosis.

The compound of the present invention has the following general formula:

Embedded image Is a chroman compound represented by Of these, R ¹ is preferably a methyl group, and R ² is preferably

Embedded image Or R ¹ is a methyl group, R ² is

Embedded image Wherein R ¹ is more preferably a methyl group,
R ² is-(CH ₂ ) ₃ -OH or a compound in which R ¹ and R ² are each represented by a methyl group. These compounds can be obtained by purchasing a commercially available product, or can be obtained by a known synthesis method. Experimental examples showing the efficacy of the present compound as a therapeutic drug for arteriosclerosis are specifically described below.

Experimental method 1) When AAPH was used as a radical initiator: 0.25 mg / ml LDL (100 μl EDTA)
M) (containing 8 mM, pH 7.4) and 2 mM of 2,
40 μl of an ethanol solution of 2,5,7,8-pentamethyl-6-chromanol (hereinafter abbreviated as PMC) or probucol and 80 μl of a 300 mM aqueous AAPH solution were added, and the final concentration was 10 μM of PMC or probucol, 3 mM of AAPH.
And The reaction was initiated by placing this solution in a 37 ° C. water bath and incubating under air. Then, 300 μl was sampled at each time. LDL used was separated from human blood by ultracentrifugation.

2) When AMVN is used as a radical initiator: 0.25 mg / ml LDL (100 μl EDTA
M) (containing 8 mM (pH 7.4)) and 2 mM of PM
40 μl of an ethanol solution of C and 20 μl of an ethanol solution of 800 mM AMVN were added to give a final concentration of 10 μM of PMC, A
MVN was 2 mM. The reaction was initiated by placing this solution in a 37 ° C. water bath and incubating under air.
Then, 300 μl was sampled at each time.

3) When copper ion is used as a radical initiator: 0.25 mg / ml of LDL (excluding EDTA) as a protein mass is adjusted to 8 ml (pH 7.4), and 20 μl of 2 mM PMC or ethanol solution of probucol is added thereto. The final concentration was 5 μM. This solution was placed in a 37 ° C. water bath and incubated under air. After 50 minutes, 2 mM
20 μl of a CuCl ₂ solution was added (final concentration 5 μM) to initiate oxidation. Then, 300 μl was sampled at each time, and the reaction was stopped by adding 3 μl of a 1 mM EDTA solution, followed by analysis.

Analysis method Chloroform / methanol = 2 /
1 solution was added to 600 μl, shaken with a vortex mixer for 1 minute, and then centrifuged at 12,000 rpm for 3 minutes with a centrifuge, and the lower layer was injected into 50 μl HPLC, and PC-OO
The concentrations of H and CE-OOH were measured.

HPLC conditions PC-OOH column: Supelco (trademark) LC-Si 2 cm + Supelco
(Trademark) LC-Si 15cm Mobile phase: MeOH / t-BuOH / 40mM phosphate = 60/30/10 Flow rate: 1.0ml / min Detection: UV 234nm Retention time: 5.6min

CE-OOH column: Supelco (trademark) LC-8 2 cm + Supelco (trademark) LC8 25 cm Mobile phase: MeOH / t-BuOH 95/5 Flow rate: 1.0 ml / min Detection: UV 234 nm Retention time: 8.2 min (20: 4CE-00H) 9.2min (18: 2CE-OOH)

Results Tables 1 to 3 show the formation of PC-OOH and CE-OOH when AAPH, AMVN and copper ions were used as radical initiators.

[Table 1]

[Table 2]

[Table 3] From Table 1, when AAPH is used as the radical initiator, P
It is clear that the addition of MC suppresses the production of PC-OOH and CE-OOH more than when nothing is added and when probucol is added. From Table 2, it is clear that even when AMVN is used as the radical initiator, the addition of PMC suppresses the production of PC-OOH and CE-OOH more than the case where nothing is added.

According to Table 3, when the copper ion was used as the radical initiator, the production of PC-OOH was suppressed when PMC was added as compared with the case where no PMC was added, and the suppressing power of PMC was superior to the case where probucol was added. . The ability to inhibit the production of CE-OOH was also superior for PMC than for probucol. From the above, it is clear that PMC suppresses the oxidation of LDL, and its action is stronger than probucol, which is currently used as an antihyperlipidemic drug. Oxidized LD
Since L is taken into macrophages and is known to cause arteriosclerosis, PMC is considered to be a potent therapeutic and preventive agent for arteriosclerosis. Further, Compound 1
Tables 4 and 5 show the production of PC-OOH and CE-OOH when AAPH and copper ions were used as the radical initiators for Nos. 5 to 5, respectively.

Embedded image

[Table 4]

[Table 5] From Tables 4 and 5, all compounds were PC-O
It is clear that it suppresses the production of OH and CE-OOH, and is considered to be useful as an agent for treating or preventing arteriosclerosis. Among them, the action of compound 1 was strong.

When the compound of the present invention is administered as a therapeutic or prophylactic agent for arteriosclerosis, it may be orally administered as tablets, granules, powders, capsules, etc., or may be administered as a suppository, injection, or external use. It may be administered parenterally as an agent.
The dosage varies significantly depending on the severity of symptoms, age, etc.
Usually, about 0.1 mg to 5000 mg, preferably 2 mg to 1000 mg, is administered once a day or several times a day for an adult.

At the time of formulation, it is produced by a conventional method using a usual formulation aid. That is, when an oral solid preparation is produced, lactose, mannitol, cornstar,
H, excipients such as crystalline cellulose, gum arabic, hydroxypropyl cellulose, a binder such as polyvinylpyrrolidone, a lubricant such as stearic acid, and, if necessary, a coloring agent or a flavoring agent, It can be manufactured by usual formulation means. When preparing an injection, a solubilizing agent, a stabilizing agent, a buffering agent and the like may be added to the main drug, if necessary, to give a subcutaneous, intramuscular, or intravenous injection by a usual method.

Embedded image

Embedded image

Claims (3)

(57) [Claims] 1. A compound of the general formula A therapeutic / prophylactic agent for arteriosclerosis containing a chroman compound represented by the formula: 2. The therapeutic or preventive agent for arteriosclerosis according to claim 1, wherein R ¹ is a methyl group and R ² is a methyl group. 3. R ¹ is a methyl group and R ² is — (CH ₂ ) ₃ —.
The therapeutic / prophylactic agent for arteriosclerosis according to claim 1, which is OH.