JPS6368550A - Novel dopamine derivative and blood platelet aggregation-inhibiting agent and 5-lipoxygenaseinhibitor containing said derivative as active ingredient - Google Patents

Abstract

NEW MATERIAL:A compound expressed by the formula (R is undecyloyl, stearoyl, oleoyl, linoleoyl or linolenoyl). EXAMPLE:N-Undecyloyl dopamine. USE:A blood platelet aggregation-inhibiting agent and 5-lipoxygenase inhibitor. Useful for remedy for cerebral infarction, cerebral thrombosis, arteriosclerosis, stenocardia, bronchial asthma and allergic disorder. Low in toxicity and high in safety. PREPARATION:A fatty acid carboxylic acid expressed by the formula RCOOH is reacted with dopamine hydrochloride in a solvent such as THF, etc., in the presence of a base (e.g. triethyl amine) and a peptide synthetic reagent (e.g. isobutyl chlorocarbonate) at 0 deg.C - room temperature to provide a compound expressed by the formula.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a novel dopamine derivative having platelet aggregation inhibiting activity and 5-lipoxygenase inhibiting activity.

[Prior art and problems] In recent years, with changes in dietary habits and aging of the population in Japan, the rapid increase in thrombotic diseases such as myocardial infarction and cerebral thrombosis has become a major social problem.

Therefore, therapeutic agents for this thrombotic disease have been investigated and developed from all aspects of their pharmacological action.

Furthermore, the number of patients with allergic diseases such as bronchial asthma is increasing, and drugs having anti-allergic effects are being searched for and developed.

[Means for Solving the Problems] The present inventors have developed a compound that has a platelet aggregation inhibiting effect that is effective in the treatment of thrombotic diseases, and a compound that has a 5-lipoxygenase inhibitory effect that is effective in the treatment of allergic diseases. As a result of intensive research, they discovered a compound represented by the following formula and completed the present invention.

That is, the compound of the present invention is a novel dopamine derivative (hereinafter referred to as a compound of the formula) represented by the following formula (wherein R represents an undecyl group, a stearoyl group, an oleoyl group, a lyloyl group, or a lynoyl group). , a platelet aggregation inhibitor and a 5-lipoxygenase inhibitor containing the derivative as an active ingredient.

In the formula, the structural formula of the undecyl group that is R is c o (
c Ht:)e CH3, the structural formula of the stearoyl group is -GO(CHz)+sCHs, and the structural formula of the oleoyl group is -GO(CH?-CH= CH-(CHt)
7c H3, the structural formula of the lyluoyl group is ~CO(CH*)
t-CH-CH-CHt-CH=CH-(CHz)-C
Ht, the structural formula of lilnoyl group is 10 0 (CH*)t
-CH-CH-CHz-CH= CH-CHt-CH-
CH-CHt CHs.

The compound of the formula can be obtained, for example, by reacting an aliphatic carboxylic acid represented by RCOOH (R has the same meaning as above) with dopamine hydrochloride in an organic solvent in the presence of a base and a peptide synthesis reagent. Can be done.

Examples of the organic solvent used include ethers such as tetrahydrofuran, diethyl ether, 1,2-dimethoxyethane, and diethylene glycol dimethyl ether, and specific examples of the base include triethylamine. Examples of bebutide synthesis reagents include isobutyl chlorocarbonate. The appropriate reaction temperature is from 0°C to room temperature, and after the reaction is complete, the compound of the formula can be purified by a combination of commonly used purification techniques such as extraction, drying, solvent removal, column chromatography, and recrystallization. can.

Next, it will be explained with reference to experimental examples that the compound of the formula of the present invention has platelet aggregation inhibiting action and 5-lipoxygenase.

[Experimental Example 1] Preparation of Platelet-Rich Plasma Nine volumes of arterial blood were collected from the femoral artery of a rabbit's lower limb into a polypropylene syringe containing one volume of 3.8% sodium citrate. The collected blood was heated to 801°C at room temperature.
) rpm for 10 minutes, and the supernatant was collected as platelet-rich plasma.

PRP). 3,000rp111 of this residue
．． Centrifuge for 15 minutes and transfer the supernatant to platelet-poor plasma (plate,
It was obtained as let-poor-plasma, P P P ). PRP was diluted using this PPP, and the platelet count was adjusted to 2.5 to 3.2×IO'/mm3.

■Measurement of platelet aggregation ability with arachidonic acid To PRPo, 4Id obtained as above, 507J of a solution of the compound of the formula obtained in Production Examples 1 to 5 described below was added and incubated at 37°C for 2 minutes. Arachidonic acid solution (final concentration 4
0μ97d) was added to induce aggregation, and the agglutination ability was measured. As a control, instead of adding the compound of formula 2
% ethanol-physiological saline was used. Measurement is PAYT
ONAGGREGATION MODULE
EL 600B, PAYTONASSOCIAT
ES) by turbidimetry, the transmittance at the time of maximum aggregation was measured, and the aggregation inhibition rate was determined when the control was set as 100%. The results are shown in Table 1.

Table 1 [Experimental Example 2] Measurement of aggregation ability using collagen Table 2 shows the results of measurements carried out in the same manner as above except that a collagen solution (final concentration 20 μ97 yd) was added instead of the arachidonic acid solution.

Table 2 [Experimental Example 3 - 5-Lipoxygenase Inhibition Effect RBL-1 cultured cells were prepared at 2 x 10'' cells/-, centrifuged at 400 x G for 10 minutes, and the residue was separated into 1 m
The cells were resuspended in a pH 7.0 phosphate buffer containing MEDTA and 0.1% gelatin to give 5 x 107 cells/-. This floating liquid was treated with ultrasonic waves to give 10,000
The supernatant obtained by centrifugation at xG for 10 minutes was used as a 5-lipoxygenase enzyme preparation.

The total volume of the reaction was 0.67, 29mM phosphate buffer, 1.
5mM calcium chloride, 10u1. Indomethacin, [
Acetone solutions of I40 core arachidonic acid, the enzyme preparation obtained as described above, and the compounds obtained in the production examples at each concentration were placed in test tubes and reacted at 37° C. for 5 minutes. After the reaction was completed, 1.2-J of acetone and 2N formic acid + 007J were added, and 1.8-J of chloroform was added. Extracted twice with d. After concentrating this extract, silica gel thin layer chromatography (developing solvent: ethyl acetate: isooctane: acetic acid: water = l I
:'5:2:10) to separate the part corresponding to the 5-lipoxygenase product [+'C]-5-HE'rE, and its radioactivity was measured using a liquid scintillation counter. . When treated in the same manner as above using an acetone solution that does not contain the compound of formula
]-5-HETE was taken as 100%, and the 50% inhibitory concentration (rc, .) of each compound was determined. The result is the third
Shown in the table.

Table 3 From these results, it was confirmed that the compound of the formula of the present invention has excellent platelet aggregation inhibiting activity and 5-lipoxygenase inhibiting activity.

Next, an acute toxicity test of the compound of the formula of the present invention was conducted using ddY male mice, and LD. All values were 1000 mg/kg or more when administered orally.

Thus, the compounds of the formula of the present invention have low toxicity and high safety.

Furthermore, in consideration of the above experimental results, the effective dose of the platelet aggregation inhibitor and 5-lipoxygenase inhibitor of the present invention that is recognized to be appropriate is the usual adult daily dose of the compound of the formula: 5-3 (1+9, I by oral administration)
It seems preferable to administer θ~100u in several doses.

Compounds of the formula of the invention can be prepared in a suitable solvent used in the formulation;
Using excipients, adjuvants, etc., preparations such as solutions, powders, granules, tablets, enteric-coated preparations, and capsules can be prepared according to conventional methods for manufacturing preparations.

In the formulation, it can also be combined with other medically active ingredients.

For oral administration, tablets, pills, capsules, powders,
It can be formulated into granules, etc.

In addition to the above-mentioned excipients, this type of preparation may contain, for example, lubricants such as magnesium stearate, sodium lauryl sulfate, and talc, binders such as dextrin, crystalline cellulose, polyvinylpyrrolidone, gum arabic, corn starch, and gelatin. Disintegrants such as sodium cellulose gelcholate, calcium cellulose gelcholate, potato starch, and carboxymethyl cellulose, and fluidity promoters such as light silicic anhydride can be used.

The drug of the present invention can also be administered as a suspension, emulsion, syrup, or elixir, and these various drug layers may contain flavoring agents, coloring agents, and the like.

The compound of the formula of the present invention has a platelet aggregation inhibitory effect and is useful for treating cerebral infarction, cerebral thrombosis, arteriosclerosis, angina pectoris, and the like.

Furthermore, since it has a 5-lipoxygenase inhibitory effect, it is also useful for treating allergic diseases such as bronchial asthma, allergic rhinitis, and atopic dermatitis.

[Example] Next, an example of manufacturing a compound of the formula of the present invention will be shown, but the present invention is not limited thereto in any way.

Production Example 1 3.729 grams of undecylic acid was dissolved in 50 grams of tetrahydrofuran under water cooling, and 2.79 grams of triethylamine was added thereto. Next, isobutyl chlorocarbonate 2.63-
was gradually added and stirred for about 15 minutes. Furthermore, 3.06 g of dopamine hydrochloride and 5.58 g of triethylamine were added to 50 g of tetrahydrofuran. Gradually add the ingredients added to d,
The mixture was stirred for about 1 hour under water cooling and further stirred at room temperature overnight. This was extracted twice with ethyl acetate, washed successively with 4% aqueous sodium hydrogen carbonate solution, water, 2% hydrochloric acid, water, and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed. The residue was subjected to silica gel column chromatography (chloroform-methanol) and recrystallized from a mixed solvent of benzene and hexane to obtain 2.369 of N-undecyl dopamine.

Melting point = 75~77℃ Infrared absorption spectrum ν, 2”x (near 1-'
:3320.2900,2840,1520°1353
．． 850,807 Proton nuclear magnetic resonance spectrum (δppm in CDC13): 0.86 (3H,t, J = 6.0 Hz).

1.24 (14H, s).

1.4-1.7 (2H, m).

2.16 (2H, t, J=7.0Hz).

2.65 (2H, t, J = 6.6 Hz).

3.44 (2H, dt, J = 6.6 Hz).

5.96 (IH, t, J = 6.6 Hz).

6.52 (IH, dd, J = 8.2 Hz).

6.74 (I H,d, J = 2.0 Hz)
.

6.80 (IH, d, J=8.0Hz) Mass spectrum: M/Z 321 (M”), 186. 13
6 Production Example 2 2.859 d of stearic acid was dissolved in 507 d of tetrahydrofuran under water cooling, and 1.39 d of triethylamine was added thereto. Next, isobutyl chlorocarbonate 1.32-
was gradually added and stirred for about 15 minutes. Furthermore, 3 g of dopamine hydrochloride, s and 2.79 g of triethylamine in tetrahydrofuran 25R1 were gradually added,
The mixture was stirred for about 1 hour under water cooling and further stirred at room temperature overnight. This was extracted twice with ethyl acetate, washed successively with 4% aqueous sodium hydrogen carbonate solution, water, 2% hydrochloric acid, water, and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed. The residue was subjected to silica gel column chromatography (chloroform-methanol) and recrystallized from a mixed solvent of benzene and hexane to obtain 1.139 of N-stearoyldopamine.

Melting point: 96. O~98.5℃ Infrared absorption spectrum::Hc71! -1:3270
．． 2985, 2835.1520°1365.858,
805 Proton nuclear magnetic resonance spectrum (δppm in CD30D): 0.89(3)i,t,J=6.0Hz).

1.29 (28H, s), 1.5 (2H, s).

2.13 (2H, bt, J=7.0Hz).

2.60 (2H, bt, J = 7.5 Hz).

3.32 (IH, t, J=7.5Hz).

6.49 (IH, dd, J=8.0, 2.2Hz).

6.63 (LH, d, J=2.2Hz).

6.67 (IH, d, J = 8.0H2) mass spectrum; M/Z 419 (M”), 284. 13
6 Production Example 3 5.649 oleic acid was mixed with 5.649 oleic acid in tetrahydrofuran under water cooling.
01111 and triethylamine 2.79
- was added. Next, isobutyl chlorocarbonate 2.6
37 was gradually added and stirred for about 15 minutes. In addition, 3.06 g of dopamine hydrochloride and 5.58 g of triethylamine
was added to 50 Id of tetrahydrofuran, and the mixture was stirred for about 1 hour under water cooling, and then stirred overnight at room temperature. This was extracted twice with ethyl acetate, washed successively with 4% aqueous sodium hydrogen carbonate solution, water, 2% hydrochloric acid, water, and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed. The residue was subjected to silica gel column chromatography (chloroform-methanol) and recrystallized from a mixed solvent of benzene and hexane to obtain 3.539 of N-oleoyldopamine.

Melting point: 50.5-52.0°C Infrared absorption spectrum t-1;333
0.2920, 2850, 1640°1530.852
,808 Proton nuclear magnetic resonance spectrum (δppm in CDC13): 0.84 (3H, t, J=6.0Hz).

1.26 (20H, s).

1.53 (2H, bt, J = 7.0 Hz).

1.97 (4H, bd, J = 5.0 Hz).

2.11 (2H, t, J=7.0Hz).

2.60 (2H, bt, J = 6.0 Hz).

3.35 (2H, bdt, J=6.6Hz).

5, 3 3 (2H, t, J =
5. 0 Hz).

6.12 (IH, bt, J=6.0Hz).

6.49 (IH, dd, J=8.0, 15.0Hz).

6.71 (IH, d, J = 15.0Hz) 6.78 (I
H, d, J = 8.0 Hz) Mass spectrum: M/Z 417 (M”), 282. 136 Production example 4 Linoleic acid 5.619 was dissolved in tetrahydrofuran 5 under water cooling.
07, and 2.7977 ml of triethylamine was added thereto. Next, isobutyl chlorocarbonate 2.63-
was gradually added and stirred for about 15 minutes. Further, 3.06 g of dopamine hydrochloride and 5.58) d of triethylamine in 507 d of tetrahydrofuran were gradually added, and the mixture was stirred for about 1 hour under water cooling and then overnight at room temperature. This was extracted twice with ethyl acetate, washed successively with 4% aqueous sodium hydrogen carbonate solution, water, 2% hydrochloric acid, water, and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed. The residue was subjected to silica gel column chromatography (chloroform-methanol) and recrystallized from a mixed solvent of benzene and hexane to obtain N-liluoyldopamine 2.079.

Properties: Colorless powder infrared absorption spectrum ν: 2SC711-': 3360
．． 2920, 2850. +640°1520.860
,820 Proton nuclear magnetic resonance spectrum (δppm in CDCl5): 0.88 (3H,t, J = 6.0 Hz).

1.27 (14H, s).

1.55 (2H, bt, J=6.0Hz).

2.05 (4H, bd, J = 6.0 Hz).

2.17 (2H, t, J=6.0Hz).

2.67 (2H, t, J=6.0Hz).

2.76 (2H, t, J=6.0Hz).

3.46 (2H, dt, J = 6.0.6.0H
z).

5.34 (4H, m).

5.86 (IH, t, J = 6.0 Hz).

6.53 (IH, dd, J=8.0, 2.0Hz).

6.74 (LH, d, J=2.0Hz).

6.81 (IH, d, J = 8.0 Hz) Mass spectrum: M/Z 415 (M”), 280. 136 Production example 5 Rirunic acid 5.579 was dissolved in tetrahydrofuran 50 under water cooling.
7, and 2.797 g of triethylamine was added thereto. Next, 2.637 g of isobutyl chlorocarbonate was gradually added and stirred for about 15 minutes. Further, 3.069 g of dopamine hydrochloride and 5.587 g of triethylamine in 507 d of tetrahydrofuran were gradually added, and the mixture was stirred for about 1 hour under water cooling and then overnight at room temperature. This was extracted twice with ethyl acetate, washed successively with 4% aqueous sodium hydrogen carbonate solution, water, 2% hydrochloric acid, water, and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed. The residue was subjected to silica gel column chromatography (chloroform-methanol).
and recrystallized from a mixed solvent of methanol and water to obtain N
- 2.569 liters of lilnoyldopamine were obtained.

Infrared absorption spectrum νwa:c7!1-':3320
．． 2930.2850.1'640゜1525.865
,810 Proton nuclear magnetic resonance spectrum (δppm in CDCl5): 0.96 (3H,t, J = 7.5 Hz).

1.25 (8H, s).

1. 5 5 (2H, bt, J = 7
．． 5 Hz).

2.07 (2H, t, J=6.0Hz).

2.14 (2H, t, J=7.5Hz).

2.65 (2H, t, J = 6.5 Hz).

2.79 (4H, t, J=5.0Hz).

3.44 (2H, dt, J=6.5, 6.5Hz).

5.34 (6H, ++).

5.94 (11-1, t, J = 6.5Hz).

6.51 (IH, dd, J=8.0, 2.0Hz).

6.73 (IH, d, J = 2.0 Hz).

6, s 1 (IH, d, J = 8.0 Hz) mass spectrum: M/Z 413 (M”), 278. 136 Next, examples of the use of the compound of the formula of the present invention are shown.

Example 1 0.5 g of the compound obtained in Production Example 1 was made into a fine powder, mixed with 98.5 g of lactose and 1 g of magnesium stearate, and this mixture was compressed using a single-shot slug tablet machine to form tablets with a diameter of 20+++a+, A slug tablet weighing 2.3g is made, crushed with an oscillator, sized, and sieved to give 20~
A good granule of 50 mesh particles was obtained.

This granule contains 1 g of Compound 5II obtained in Production Example 1 in 1 g, and the dose is 2 to 20 g divided into three doses per day depending on the symptoms.

Example 2 0.5 g of the compound obtained in Production Example 2 was made into a fine powder, mixed with 98.5 g of lactose and Ig of magnesium stearate, and this mixture was compressed using a single-shot slug tablet machine to form tablets with a diameter of 2011 mm and 111 mm by weight. Make 2.3g slug tablets, crush them with an oscillator, size them, and sieve them for 20~
A good granule of 50 mesh particles was obtained.

Each gram of this granule contains 5 mg of the compound obtained in Production Example 2, and the dose is 2 to 20 g divided into three doses per day depending on the symptoms.

Example 3 Microcrystalline cellulose 93 was added to 2.5 g of the compound obtained in Production Example 3.
g1 3 g of sodium cellulose gelcholate and 1.5 g of magnesium stearate were added and mixed, and this mixture was compressed using a single-shot tablet machine to produce tablets with a diameter of 911 and a weight of 200 mg.

Each tablet contains 5 mg of the compound obtained in Example 1, but the dosage is 2 to 20 tablets divided into three doses per day depending on the symptoms.

Example 4 Microcrystalline cellulose 93 was added to 2.5 g of the compound obtained in Production Example 4.
3 g of sodium g5 cellulose gelcholate and 1.5 g of magnesium stearate were added and mixed, and the mixture was compressed using a single-shot tablet machine to produce tablets with a diameter of 911 and a weight of 200 mg.

Each tablet contains 5 mg of the compound obtained in Example 2, but the dosage is 2 to 20 tablets divided into three doses per day depending on the symptoms.

Example 5 10g of the compound obtained in Production Example 5 was made into a fine powder, and this was mixed with lactose 8
6.5 g, 0.5 g of light anhydrous silicic acid, and 3 g of microcrystalline cellulose were mixed, and 10 (1+ g each) of the mixture was filled into hard capsules to obtain capsules.

This capsule contains 10 mg of the compound obtained in Production Example 5 per 1 capsule, but 1-1 mg per day is added depending on the symptoms.
Take 0 capsules in 3 doses.

Example 6 10g of the compound obtained in Production Example 2 was made into a fine powder, and this was mixed with lactose 8
6.5 g, 0.5 g of light anhydrous silicic acid, and 3 g of microcrystalline cellulose were mixed, and 100 mg each of the mixture was filled into hard capsules to obtain capsules.

This capsule contains 10n+g of the compound obtained in Production Example 2 in 1 capsule, but 1-1-g per day is added depending on the symptoms.
Take 10 capsules in 3 divided doses.

Example 7 Compound 19 obtained in Production Example 4 was dissolved in 15o1iIi polysorbate 80, 4.85C of sterile physiological saline heated to 60°C was added and shaken well, and this was aseptically produced into a vial. The compound obtained in Example 4 was distributed to contain 3ff1g and sealed to prepare an injection.

This injection should be shaken before use.
Administer intravenously for ~150d.

Example 8 Compound 19 obtained in Production Example 5 was dissolved in 1507 polysorbate 80, 4.85Q of sterile physiological saline heated to 60°C was added and shaken well, and this was aseptically placed in a vial. The compound obtained in step 5 was distributed to contain 3 mg and sealed to produce an injection.

This injection should be shaken before use.
~150 - Administer intravenously.

Claims (3)

[Claims] (1) A novel dopamine derivative represented by the following formula ▲ Numerical formula, chemical formula, table, etc.▼ (In the formula, R represents an undecyl group, a stearoyl group, an oleoyl group, a linoleoyl group, or a linolenoyl group.) (2) The active ingredient is a new dopamine derivative represented by the following formula ▲ Numerical formulas, chemical formulas, tables, etc. A platelet aggregation inhibitor. (3) The active ingredient is a new dopamine derivative represented by the following formula ▲ Numerical formulas, chemical formulas, tables, etc. 5-
Lipoxygenase inhibitor.