JPS6241500B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to an acylated indole derivative, a method for producing the same, and a platelet aggregation inhibitor containing the same as an active ingredient. The novel compound provided by the present invention, an acylated indole derivative, has excellent platelet aggregation inhibiting activity, and therefore can be used to prevent occlusion in the cardiovascular system, to prevent and treat postoperative thrombosis, and to improve blood vessel function after surgery. It is an extremely useful compound for the prevention and treatment of thromboembolism, atherosclerosis, arteriosclerosis, etc., and for the prevention and treatment of myocardial infarction and recurrence after stroke. Furthermore, the novel acylated indole derivatives provided by the present invention are expected to have excellent anti-inflammatory and fibrinolytic activity, and are extremely useful compounds. The novel acylated indole derivative provided by the present invention has the following formula [] [In the formula, R ₁ represents a hydrogen atom, a lower alkyl group or a lower acyl group, R ₂ represents a methyl group, and R ₃ and R ₄ represent a hydrogen atom. φ represents an unsubstituted aryl group. R ₅ represents a hydrogen atom or a lower alkyl group having 2 to 5 carbon atoms. ] It is expressed as . US Pat. No. 3,557,142 describes acylated indole derivatives having anti-inflammatory effects with low toxicity, and these acylated indole derivatives have a carboxyl group or an ester group as an N-substituent. Also, British Patent No.
Publication No. 959203 describes acylated indole derivatives having an antiallergic effect, and such acylated indole derivatives have an aminoalkyl group as an N-substituent. As mentioned above, the acylated indole derivative of the present invention is
The acylated indole derivative provided by the present invention, which has an alcoholic hydroxyl group, ether group, or acyloxy group in the N-substituent, is clearly different from that described in the above-mentioned publications, etc., and is The acylated indole derivative of the present invention, which is a completely unknown new compound, has a unique pharmacological action of extremely excellent platelet aggregation inhibiting action, and is also known as a compound that has anti-inflammatory action and fibrinolytic action. Therefore, it is expected to be extremely useful as a medicine. In the acylated indole derivative represented by the above formula [] provided by the present invention, R ₁ is a hydrogen atom, a lower alkyl group, or

[Preparation of PRP/drugs and flocculants]

(1) Preparation of PRP (platelet-rich plasma) Citrated blood (3.8%
1 volume of sodium citrate to 9 volumes of blood) was collected. The obtained citrate blood was centrifuged at 1000 rpm for 10 minutes at room temperature, and the supernatant (PRP) was separated. The obtained PRP was stored at room temperature and used as soon as possible, and was not used after 4 hours after conditioning. (2) Preparation of drug The test drug is generally dissolved in dimethyl sulfoxide to a concentration of 100 mg/ml, diluted with physiological saline, and prepared at 2500 μg/ml, 1000 μg/ml, and 1000 μg/ml.
750μg/ml, 500μg/ml, 250μg/ml, 100
1 ml of each μg/ml solution was prepared. However, a compound having a free carboxyl group was dissolved in 0.1M NaHCO ₃ to obtain a 10 mg/ml sodium salt solution, and similarly diluted with physiological saline to prepare a test drug solution. Platelet aggregation inhibition test result: 1000μg/ml
If the drug completely suppresses platelet aggregation even at a final concentration of 10 μg/ml, dilute the drug solution with physiological saline (75 μg/ml, 50 μg/ml). , 25μg/ml, 10
Platelet inhibition tests were also performed on each dilution (μg/ml). (3) Adjustment of flocculant Arachidonic acid (99% pure) manufactured by Sigma
Prepare 3.3mM arachidonic acid sodium salt solution by dissolving in _0.1M NaHCO3. This was diluted with physiological saline to 1mM and used in the experiment. The 3.3mM solution was stored in the refrigerator as a stock mother solution, and the 1mM solution was newly prepared from this mother solution each time it was used. [Platelet aggregation inhibition test] (1) Blank platelet aggregation 25μ
Platelets were agglutinated by adding 25μ of physiological saline and a 25μ agglutinant solution, and the aggregation curve was recorded for 3 minutes using a Priceton aggregometer. The maximum aggregation degree in this platelet aggregation was defined as the maximum aggregation degree of the blank. (2) Adjustment of coagulation strength of arachidonic acid sodium salt Add 25μ of ibuprofen solution (250μg/ml or 100μg/ml) to 200μ of PRP,
Pre-incubation was carried out in an aggregometer cuvette while stirring with a stirrer for 2 minutes in the same manner as in (1) above, and then 25μ
Add 1mM sodium arachidonic acid solution to the above
A platelet aggregation curve was recorded in the same manner as in (1), the maximum degree of aggregation was calculated from the curve, and the aggregation inhibition rate was calculated using the following formula. Inhibition rate (%) = 100 - maximum aggregation degree of ibuprofen-added system / maximum aggregation degree of blank x 100 At this time, the inhibition rate was 50% or more in the system with ibuprofen added at 250 μg/ml, and the inhibition rate was over 50% in the system added with ibuprofen at 100 μg/ml. It was confirmed that the inhibition rate was below 50%, and if it was not, the sodium arachidonic acid salt solution was not at an appropriate concentration, so the concentration was readjusted and the test was performed again to match the conditions. (3) Platelet aggregation inhibition test Add 25μ of the test drug solution to 200μ of PRP, pre-incubate at 37℃ for 2 minutes in the same manner as in (2) above, and then add arachidonic acid with the concentration adjusted in (2) above. 25μ of sodium solution was added, an aggregation curve was recorded for 3 minutes, the maximum degree of platelet aggregation within that time was measured, and the inhibition rate was calculated using the following formula. The lowest concentration of drug at which the inhibition rate exceeded 50% was expressed as the IC ₅₀ value. Inhibition rate (%) = 100 - (maximum aggregation degree of test drug added system/maximum aggregation degree of blank x 100) The results are shown in Table 1. In addition, an acute toxicity test was conducted using mice by oral administration of the test drug. The _LD50 value is
It was over 3000mg/Kg.

[Table] Example 5 Preparation of tablets Tablets were manufactured, each having the following composition. Active ingredient 200mg Lactose 280mg Gym starch 80 mg Polyvinylpyrrolidone 11mg Magnesium stearate 5mg 576mg The active ingredient, lactose and Gyote starch were mixed, evenly moistened with a 20% ethanol solution of polyvinylpyrrolidone, and passed through a 20mm mesh sieve. It was dried at 45°C and passed through a 1.5 mm mesh sieve again. The granules thus obtained were mixed with magnesium stearate and compressed into tablets. As an active ingredient, typically 3-benzoyl-1
-(1-hydroxy-2-propyl)indole was used. Example 6 Formulation of Capsules Hard gelatin capsules were prepared, one capsule containing the following composition: Active Ingredients 200mg Microcrystalline Cellulose 195mg Amorphous Silicic Acid 5mg 400mg The active ingredient in finely powdered form, microcrystalline cellulose and unpressed amorphous silicic acid were thoroughly mixed and packed into hard gelatin capsules. As an active ingredient, typically 3-benzoyl-1
-(1-hydroxy-2-propyl)indole was used. Example 7 Preparation of ampoule An ampoule containing the following composition in one ampoule (5 ml volume) was manufactured. Active ingredient 200 mg Polyethylene glycol 600 200 mg Distilled water Total volume 5.0 ml Polyethylene glycol and active ingredient were dissolved in water under nitrogen, which was boiled, cooled under nitrogen and distilled. The solution was made up to the given volume with pretreated water and filtered under sterile conditions. This production is carried out under diffused light. Filling was carried out in a nitrogen stream and sterilization was carried out at 121°C for 20 minutes. In addition, 3-benzoyl-1- as an active ingredient
(1-hydroxy-2-propyl)indole was used.

Claims (1)

[Claims] 1. The following formula [] [In the formula, R ₁ represents a hydrogen atom, a lower alkyl group or a lower acyl group, R ₂ represents a methyl group, and R ₃ and R ₄ represent a hydrogen atom. φ represents an unsubstituted aryl group. R ₅ represents a hydrogen atom or a lower alkyl group having 2 to 5 carbon atoms. ] An acylated indole derivative represented by 2. The acylated indole derivative according to claim 1, wherein φ is a phenyl group. 3. The acylated indole derivative according to claim 1 or 2, wherein R ₅ is a hydrogen atom or an iso-propyl group. 4 The following formula [] [In the formula, R ₁ represents a hydrogen atom or a lower alkyl group, R ₂ represents a methyl group, and R ₃ and R ₄ represent a hydrogen atom.
φ represents an unsubstituted aryl group. R ₅ represents a hydrogen atom or a lower alkyl group having 2 to 5 carbon atoms. ] A platelet aggregation inhibitor comprising an effective amount of an acylated indole derivative represented by the following formula and a pharmaceutically acceptable carrier.