JPH0710775B2 - Angiotensin I converting enzyme inhibitor - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to angiotensin I converting enzyme inhibitors.

Angiotensin I is the amino acid 10
Angiotensinogen, a decapeptide consisting of individual molecules, that exists in serum and has a molecular weight of about 100,000.
Is produced by the enzyme renin, which is present in the kidney. Furthermore, this Angiotensin I
It is converted into active angiotensin II by the action of the converting enzyme. This angiotensin II is an octapeptide and has a strong vasoconstrictor action,
It is said to be the main body of renal pressor substance. In addition, angiotensin I converting enzyme is a kinin (ki
It is also known to decompose and inactivate nin).
Therefore, the angiotensin I-converting enzyme inhibitor is effective in treating hypertension in that it inhibits the production of pressor substances and the decomposition of antihypertensive substances.

Therefore, the present inventors have conducted extensive studies in order to develop a drug useful for treating hypertension, and as a result, found that the compound represented by the general formula has an excellent angiotensin I converting enzyme inhibitory action, Based on this, the present invention has been completed.

That is, the present invention has the general formula (In the formula, R is hydrogen, a lower alkylcarbonyl group,
Alternatively, it represents an aromatic carbonyl group. ) Is an angiotensin I-converting enzyme inhibitor (hereinafter referred to as the agent of the present invention) containing an unsaturated fatty acid derivative represented by the following formula and a pharmacologically acceptable salt thereof as an active ingredient.

[Examples] 9-Hydroxy-10,12,15-octadecatrienoic acid Among the compounds represented by the above general formula, the compound in which R is hydrogen is 9-hydroxy-10,12,15-octadecatriene. Lycii Radici, which is an acid, for example, the Chinese medicine that is distributed in Chinese herbal medicines such as Jiin Shiho-to and Seishin-Renko
s Cortex). The skeleton is the dried root bark of the wolfberry (Lycium chinense MILLER) of the Solanaceae family, or a closely related plant.

9-Hydroxy-10,12,15-octadecatrienoic acid is
For example, the skeleton is extracted with chloroform, a mixed solvent of chloroform-methanol or acetone, the solvent is removed from the extract, the residue is dissolved in hydrated methanol, and this is extracted with n-hexane, petroleum ether, etc. After doing
The solvent is removed from the water-containing methanol layer, and the residue is subjected to general chromatography using silica gel and / or
Alternatively, it can be obtained by subjecting it to high performance liquid chromatography.

A specific example of the production of this 9-hydroxy-10,12,15-octadecatrienoic acid is shown below.

Concrete Example 1 10 kg of powdered earth and bone skin was added to and mixed with chloroform 100, extracted at room temperature, and the solvent was removed from the extract to obtain 76 g of chloroform extract. After dissolving this chloroform extract in 90% (v / v) methanol-water mixed solvent 1, transfer to a separating funnel, add n-hexane 1 and shake and stir, then take the lower layer and remove the solvent. 90% removed
18 g of (v / v) methanol-water extract was obtained. This 90% (v /
v) Methanol-water extract was subjected to column chromatography using silica gel as an adsorbent, and n-hexane,
Eluted with n-hexane-benzene, n-hexane-ether, ether-ethyl acetate, ethyl acetate-methanol, and fractionated with ether-ethyl acetate (1: 1) (about 2 g) by high performance liquid chromatography. Column, μ-Bondapak C
₁₈ (diameter 8 mm, length 30 cm); mobile phase, acetonitrile: 1% acetic acid (3: 2); flow rate, 3 ml / min] and colorless oily substance 0.1
got g.

This colorless oily substance has the following physicochemical properties and is described in the literature [HWS Chan and G. Levett Lipids 12 99 (197
7)] and the physicochemical properties of 9-hydroxy-10,12,15-octadecatrienoic acid.

Infrared absorption spectrum 3596,1708,1462,1410 Proton nuclear magnetic resonance spectrum (δppm in CDCl ₃ ) 0.98 (3H, t, J = 7Hz), 2.08 (2H, m), 2.34 (2H, t, J = 7Hz), 2.93 (2H) , dd, J = 7,7Hz), 4.17 (1H, m), 5.35 (3H, m), 5.68 (1H, dd, J = 15,7Hz), 5.98 (1H, dd, J = 11,11Hz), 6.50 (1H, dd, J = 11,15Hz) 9-acetoxy-10,12,15-octadecatrienoic acid Among the compounds represented by the above general formula, the compound in which R is an acetyl group is 9-acetoxy-10 , 12,15-Octadecatrienoic acid. To obtain this compound, the compound obtained in the above-mentioned specific example 1 is acetylated by a conventional acetylation reaction, for example, reaction with acetic anhydride-pyridine. Can be obtained.

A specific example of the production of 9-acetoxy-10,12,15-octadecatrienoic acid is shown below.

Specific Example 2 30 mg of 9-hydroxy-10,12,15-octadecatrienoic acid obtained in Specific Example 1 was dissolved in 1 ml of pyridine, and acetic anhydride 0.5
ml was added and left overnight at room temperature. 30 ml of ice water in this reaction solution
Was added and the mixture was extracted with 20 ml of ether, and the ether layer was removed of the solvent under reduced pressure to obtain 40 mg of a reaction product. This reaction product was collected by preparative medium pressure liquid chromatography [column, Kusano Kagaku Kikai Co., Ltd. CIG prepack column (diameter 15 mm, length 30 c
m); packing material, silica gel (crushed to 10 μm); mobile phase, n-
Hexane: ethyl acetate (5: 1); flow rate, 2 ml / min] to obtain 20 mg of 9-acetoxy-10,12,15-octadecadienoic acid as a colorless oily substance.

9-Benzoyloxy-10,12,15-octadecatrienoic acid Among the compounds represented by the above general formula, the compound in which R is a benzoyl group is 9-benzoyloxy-10,12,15-octadecatrienoic acid. In order to obtain this compound, the compound obtained in the above-mentioned specific example 1 can be obtained by benzoylating it by a commonly used benzoylation reaction, for example, a reaction with benzoyl chloride-triethylamine.

A specific example of the production of this 9-benzoyloxy-10,12,15-octadecatrienoic acid is shown below.

Example 3 20 mg of 9-hydroxy-10,12,15-octadecatrienoic acid obtained in Example 1 was dissolved in 4 ml of triethylamine, a small amount of dimethylamitopyridine and 0.5 ml of benzoyl chloride were added, and the mixture was stirred at room temperature under a nitrogen stream. It was stirred for 15 hours. Ice water (20 ml) was added to the reaction solution, the mixture was extracted with ether (20 ml), and the solvent was removed from the ether layer under reduced pressure to obtain a reaction product (35 mg).
This reaction product was collected by preparative medium pressure liquid chromatography (column, Kusano Kagaku Kikai Co., Ltd. CIG pre-pack column (diameter
15 mm, length 30 cm); packing material, silica gel (10 μm crushed); mobile phase, n-hexane: ethyl acetate (5: 1); flow rate, 2
ml / min] to obtain 17 mg of 9-benzoyloxy-10,12,15-octadecatrienoic acid as a colorless oily substance.

Next, the fact that the compound represented by the general formula, which is the active ingredient of the drug of the present invention, has an angiotensin I converting enzyme inhibitory action will be described with reference to experimental examples.

Experimental example 2 g of Rabituto guar cent powder (manufactured by Sigma) 50
Dissolve in 30 ml of mM phosphate buffer (pH 8.3), and add 40 at 34,000G.
Centrifuge for 10 minutes and add the supernatant to a dialysis tube.
After dialysis with mM phosphate buffer solution 3, it was further diluted 2-fold with 50 mM phosphate buffer solution to obtain an angiotensin I converting enzyme solution.

A sample containing the compound obtained in Example 1 was placed in a test tube (25 μm), and 50 μm of hypryl histidyl leucine (final concentration 5 mM) was added as a substrate to the test tube. (PH 8.3) 50 μl was added and incubated at 37 ° C for 10 minutes, then the enzyme solution obtained as above
100 μm was added, and the mixture was reacted at 37 ° C. for 60 minutes. After that, 100 μ of 1N hydrochloric acid was added to stop the reaction, and then 25 μ of benzoylalanine (1 mg / ml) was added as an internal standard, and 1 ml was added.
Ethyl acetate was added, and the amount of hypuric acid extracted in ethyl acetate was measured by high performance liquid chromatography [column, μ-Bondapak C ₁₈ (diameter 4 mm, length 30 cm); mobile phase, acetonitrile: methanol: 1% Acetic acid (1: 1: 8); Flow rate, 1m
l / min; detection, ultraviolet ray (254 nm)], and used as the enzyme activity.

With respect to this result, the inhibition rate was calculated by the following formula.

C: Hypuric acid peak area when compound not obtained in specific example (corrected by internal standard) S: Hypuric acid peak area when compound obtained in specific example was added (corrected by internal standard) IC _{When 50} was determined, it was 0.9 mM, and an obvious angiotensin I converting enzyme inhibitory action was recognized.

Next, an acute toxicity test of the compounds obtained in Specific Examples 1 to 3, which are active ingredients of the drug of the present invention, was carried out using ddY mice, and both were dead cases by oral and intraperitoneal administration of 2 g / kg. It was a long time ago.

As described above, the compound represented by the general formula has extremely low toxicity and high safety.

In addition, the compound represented by the general formula can also be used as a commonly used pharmaceutical salt such as sodium, potassium, calcium, aluminum, ammonium, diethylamine, triethanolamine, etc., in order to achieve the intended effect. .

Next, the dose and formulation of the drug of the present invention will be described.

The compound represented by the general formula can be administered to animals and humans as it is or together with a conventional pharmaceutical carrier. The dosage form is not particularly limited and may be appropriately selected and used as necessary. Oral preparations such as tablets, capsules and granules,
Parenteral agents such as injections and suppositories can be mentioned.

In order to exert the intended effect as an oral agent, it depends on the age, weight and degree of disease of the patient, but usually 0.5 to 1.0 g / day is used as the weight of the compound represented by the general formula in adults.
It seems appropriate to take it in divided doses.

In the present invention, oral preparations such as tablets, capsules and granules are manufactured according to a conventional method. Tablets are produced by mixing the compound represented by the general formula with a pharmaceutical excipient such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic, etc. It is manufactured by mixing with an inert formulation filler or a diluent and filling it into a hard gelatin capsule, a soft gelatin capsule or the like. Syrups and elixirs are
A sweetener such as sucrose, a preservative such as methyl and propylparabens, a coloring agent, a seasoning, a compound represented by the general formula:
It is manufactured by mixing with fragrance and auxiliary agents.

In order to exert a desired effect as a parenteral agent, it depends on the patient's age, body weight, and degree of disease, but usually, in adults, the daily dose of the compound represented by the general formula is from 50 to 300 mg per day. Injection, subcutaneous injection, and intramuscular injection seem appropriate.

This parenteral preparation is produced by a conventional method, and distilled water for injection, physiological saline, dextrose aqueous solution, propylene glycol or the like can be generally used as a diluent. Further, if necessary, a bactericide, a preservative, and a stabilizer may be added. Further, from the viewpoint of stability, this parenteral preparation may be filled in a capsule or the like and then frozen, the water content may be removed by an ordinary freeze-drying technique, and a liquid preparation may be re-prepared from the freeze-dried product immediately before use.

Other parenteral agents include liquid preparations for external use, coating agents such as ointments, suppositories for rectal administration, and the like, and they are manufactured by a conventional method.

Next, the present invention will be specifically described with reference to examples, but the present invention is not limited thereto.

Example 1 200 g of the drug obtained in Example 1 was dissolved in 150 ml of polysorbate 80, and sterilized physiological saline 4.85 heated to 60 ° C. was added to the solution.
Was thoroughly shaken, and the mixture was aseptically distributed into vials so that the drug obtained in Example 1 contained 300 mg, and the mixture was sealed to produce an injection.

This injection is shaken at the time of use and the daily dose is 1.5 ~ depending on the symptoms.
Administer 7.5 ml intravenously.

Example 2 100 g of the drug obtained in Example 2 was mixed with 20 g of silicic acid anhydride, to which 75 g of corn starch was added and further mixed. To this mixture was added 100 ml of 10% hydroxypropylcellulose / ethanol solution, and the mixture was kneaded in the usual manner, extruded, dried and sieved to obtain granules of 20 to 50 mesh particles.

This granule is to be taken 3 times a day in a dose of 0.4 to 0.6 g (corresponding to 0.2 to 0.3 g as the weight of the drug obtained in Example 2) according to the symptoms.

Example 3 20 g of the drug obtained in Example 3 was mixed with 20 g of silicic acid anhydride, and 10 g of microcrystalline cellulose and 0.5 g of magnesium stearate were added to the mixture.
g and lactose 49.5 g were added and mixed, and this mixture was tabletted by a single-shot tableting machine to produce a tablet having a diameter of 7 mm and a weight of 100 mg.

One tablet of the present invention contains 100 mg of the drug obtained in Example 3.
This tablet is taken 2-3 times at a time, three times a day.

Example 4 170 mg of the drug obtained in Example 2 was mixed with 100 mg of lactose and filled in a No. 0 gelatin capsule to obtain a capsule.

This capsule is taken 1 to 2 capsules 3 times a day depending on the symptoms.

 ─────────────────────────────────────────────────── --- Continuation of the front page (72) Inventor Hiroshi Mitsuhashi 1-3-5 Minami-Kashiwa, Kashiwa-shi, Chiba Condominium Minami-Kashiwa 606 Examiner, Shoei Hoshino

Claims (1)

[Claims] 1. A general formula (In the formula, R is hydrogen, a lower alkylcarbonyl group,
Alternatively, it represents an aromatic carbonyl group. ) An angiotensin I-converting enzyme inhibitor containing an unsaturated fatty acid derivative represented by the following formula and a pharmacologically acceptable salt thereof as an active ingredient.