JP2811608B2 - Phosphonamide derivatives and endothelin converting enzyme inhibitors - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel phosphonamide derivative and an endothelin converting enzyme inhibitor using the same.
Agent .

[0002]

2. Description of the Related Art Endothelin (ET-1), a peptide consisting of 21 amino acid residues isolated from a culture supernatant of porcine aortic endothelial cells, endothelium-independently regulates various arteries such as coronary arteries and pulmonary arteries. It is known that the vasoactive substance is an extremely powerful and persistent vasoactive substance having an EC ₅₀ of about 4 × 10 ⁻¹⁰ M for coronary artery contraction. E
Since T-1 has a strong vascular smooth muscle contracting action and a blood pressure increasing action, ET-1 is considered to be deeply involved in the onset of hypertension.

Processing of ET-1 precursor in vivo
Produces big endothelin (big ET-1).
The vasoconstrictor activity of big ET-1 is about 1 / that of ET-1.
The blood pressure is 100 when big ET-1 is administered to a living body.
Is increased, big ET-1 is converted to endothelin
Under the action of an enzyme (ECE), ET-1 is produced, and ET-1 is produced.
It is considered that the blood pressure increases due to the action of -1. big E
Before administering T-1 to a living body, a metal protease inhibitor
Administering phosphoramidon, known as
Since the rise in blood pressure is suppressed, phosphoramidon is
ET-1 to inhibit the action of endothelin converting enzyme
Production is suppressed, and thus the rise in blood pressure is suppressed
(European Journal of Pharmacology,18
Five, 103-106 (1990)).

[0004]

SUMMARY OF THE INVENTION Action to inhibit endothelin converting enzyme activity (endothelin converting enzyme inhibitory activity)
To provide a novel substance which is strong and has an excellent blood pressure lowering action.

[0005]

Means for Solving the Problems The present invention provides the following formula (1)

[0006]

Embedded image

Wherein A represents an amino acid residue selected from the group consisting of leucine, isoleucine and phenylalanine; Trp represents a tryptophan residue;
May be the same or different and represent a lower alkyl group, a lower alkoxy group, or a halogen atom, X represents a hydrogen atom or an alkali metal, Y represents a hydrogen atom, an alkali metal or a lower alkyl group, and m represents is an integer of 0 to 3, n is used phosphonic acid amide derivatives and their characterized in that it is represented by a is) integer from 1 to 4
An endothelin converting enzyme inhibitor is provided .

In the above formula (1), R is preferably a lower alkyl group having 1 to 4 carbon atoms (particularly methyl and ethyl), a lower alkoxy group having 1 to 4 carbon atoms (particularly methoxy and ethoxy). Or a halogen atom such as chlorine or fluorine. As the alkali metal represented by X and Y, sodium and potassium are preferable. As the lower alkyl group represented by Y, a lower alkyl group having 1 to 4 carbon atoms, particularly methyl, ethyl and isopropyl is preferable. m is preferably 0 or 1, and particularly preferably 0 (that is, the phenyl group in Formula (1) has no substituent).

[0009] Among the phosphonamide derivatives of the present invention,
Particularly preferred compounds are exemplified below. (A) Ph-CH ₂ -P (O) (ONa) -Leu-Trp-ONa (B) Ph- (CH ₂ ) ₂ -P (O) (ONa) -Leu-Trp-ONa (C) Ph- (CH ₂ ) ₂ -P (O) (ONa) -Ile-Trp-ONa (D) Ph- (CH ₂ ) ₂ -P (O) (ONa) -Ile-Trp-OCH ₃ (E) Ph- ( CH ₂ ) ₂ -P (O) (ONa) -Phe-Trp-ONa (F) Ph- (CH ₂ ) ₄ -P (O) (ONa) -Leu-Trp-ONa (G) Ph- (CH ₂ ) ₄ -P (O) (ONa) -Ile-Trp-ONa (H) Ph- (CH ₂ ) ₂ -P (O) (OH) -Leu-Trp-OH (I) Ph- (CH ₂ ) ₂ -P (O) (OH) -Ile-Trp-OH

In the above formulas, Ph represents a phenyl group, Leu represents an L-leucine residue, and Ile represents L-leucine.
-Represents an isoleucine residue, Phe represents an L-phenylalanine residue, and Trp represents an L-tryptophan residue.

Among the above compounds, the compounds (A), (B) and (F) are common in that they contain a phosphoramidone and a Leu-Trp group, but the phosphoramidone has a glycosyl group. In contrast, the compounds of the present invention differ in having a phenylalkylene group, and the compounds of the present invention are completely different from phosphoramidone.

Next, the endothelin converting enzyme inhibitory activity of the compound of the present invention will be described. The endothelin converting enzyme inhibitory activity was measured as follows.

[Extraction of endothelin converting enzyme]
A 0-260 g male Wistar rat was bled by jugular vein transection, and the lungs were removed. Bronchi were excised from the rat lung in ice-cold 10 mM phosphate buffered saline (pH 7.2), and then in a 10 mM Tris-HCl buffer (pH 7.4), a Teflon homogenizer at 1000 rpm for three times. After crushing by moving, the residue was removed by centrifugation at 1800 × g for 20 minutes. Subsequently, the mixture was centrifuged at 30,000 × g for 90 minutes to obtain a membrane fraction as a precipitate. This precipitate was suspended in a 10 mM Tris-HCl buffer (pH 7.4) containing 0.5% Triton X-100, and then homogenized with a Teflon homogenizer at 1000 rpm and twice up and down to solubilize. This was centrifuged at 30,000 × g for 90 minutes, and the supernatant was used as a membrane extraction fraction. This fraction was subjected to anion exchange column chromatography (DE52, Whatman), and eluted with a concentration gradient of 0-0.5M sodium chloride.
The endothelin-converting activity of each of the obtained fractions was detected by the measurement method described below, and the fractions having the endothelin-converting activity were pooled.

[Endothelin converting enzyme activity] Enzyme activity measurement 10 μl (10 pmol) of pig big ET-1 as a substrate and 0.1 M Tris-HCl buffer (pH 7.0) were
Put into a reaction tube to a final volume of 125 μl,
100 μl of the fraction having endothelin conversion activity obtained as described above was added to start the reaction. 37 ° C, 3
After incubation for 1 hour, the reaction was stopped by placing in boiling water for 1 minute, and centrifuged at 1500 × g for 10 minutes to obtain a supernatant. The amount of ET-1 produced in the supernatant was
Measured by T-1 specific radioimmunoassay.

2. Enzyme inhibitory activity The endothelin converting enzyme inhibitory activity was determined by measuring the endothelin converting enzyme activity in the presence and absence of a sample, and expressed as the inhibition rate. That is, at the beginning of the enzyme activity measurement, a sample dissolved in a 0.1 M Tris-HCl buffer (pH 7.0) was added to a reaction test tube to a final concentration of 10 ⁻⁴ M, and an enzyme reaction was performed.

The inhibition rate was determined by measuring the ET-1 in the reaction solution containing the sample.
The amount (a), the amount of ET-1 in the reaction solution containing no sample (b), and the background of the test tube in which 0.1 M Tris-HCl buffer (pH 7.0) was added instead of the enzyme solution. E
It was determined from the T-1 amount (c) by the following equation (A).

Inhibition rate = [(ba) / (bc)] × 100 (%) (A)

The inhibitory rates of the phosphonamide derivatives obtained in the following Examples and Comparative Examples against the endothelin converting enzyme activity of the rat lung membrane fraction were as shown in Table 1. Table 1 also shows the inhibitory rates of phosphoramidon obtained in the same manner.

[0019]

As is clear from the results in Table 1, the compounds of the present invention, which are obtained in each of the examples, exhibit endothelin converting enzyme inhibitory activity equivalent to that of phosphoramidone, but are outside the scope of the present invention. The compound obtained in Comparative Example 1 has remarkably weak endothelin converting enzyme inhibitory activity.

The phosphonamide derivative of the present invention has a strong inhibitory activity on endothelin converting enzyme activity, and has essential hypertension, acute heart failure, renal failure, cerebral vasospasm (after subarachnoid hemorrhage), It has pharmacological effects on preeclampsia, coronary spasm, myocardial infarction, diabetes, asthmatic attacks, Raynaud's syndrome, etc.

REFERENCE EXAMPLE 1 a) Dibenzyl 4-phenylbutylphosphonate (R1a) Under a nitrogen atmosphere, 6.15 g of dibenzyl phosphite (2
5 mmol) of dimethylformamide (DMF) (40
ml) solution was cooled to -15 ° C, 1.10 g (27.5 mmol) of 60% sodium hydride was added, and the mixture was stirred at 0 ° C or lower for 1.5 hours. Next, a solution of 5.0 g (25 mmol) of 1-bromo-4-phenylbutane in DMF (5 ml) was added dropwise so that the temperature of the reaction solution did not rise to 0 ° C. or higher. The mixture was stirred at room temperature overnight, and the reaction solution was concentrated under reduced pressure. The residue was dissolved in ether, washed with water and dried over anhydrous sodium sulfate. From this, the solvent was distilled off to obtain a colorless oil. This was purified by silica gel chromatography to give the title compound (6.90 g, yield 70.0%) as a colorless oil.

B) Dibenzyl 2-phenethylphosphonate (R1b) 1.31 g (5.0 mmol) of dibenzyl phosphite
And 0.93 g of 1-bromo-2-phenylethane (5.
0 mmol), and synthesized in the same manner as in the synthesis of compound (R1a) to give 1.11 g of the title compound (yield; 60.7%).
%).

C) Dibenzylbenzylphosphonate (R1c) Same as the synthesis of compound (R1a) from 2.64 g (10.0 mmol) of dibenzylphosphite and 1.26 g (10.0 mmol) of benzyl bromide. To give 0.93 g (yield; 26%) of the title compound.

Reference Example 2 a) Benzyl 4-phenylbutylphosphonochloride (R2a) A solution of 3.0 g (7.6 mmol) of compound (R1a) in carbon tetrachloride (4.6 ml) was cooled with ice. Under phosphorus pentachloride 1.6
6 g (8.0 mmol) was added and stirred for 30 minutes. The reaction temperature was raised to 70 ° C at a rate of 10 ° C / 15 minutes,
Stirred at 30 ° C. for 30 minutes. The reaction solution was placed under reduced pressure (1 mmH
g) The mixture was kept at 70 ° C. for 2 hours, and the solvent and by-products (phosphorus oxychloride, benzyl chloride) were distilled off under reduced pressure. This was used for the next reaction without further purification.

B) Benzyl 2-phenethylphosphonochloride (R2b) Compound (R1b) was synthesized from 2.10 g (5.73 mmol) of compound (R1b) in the same manner as in the synthesis of compound (R2a).

C) Benzyl Benzylphosphonochloride (R2c) Compound (R1c) was synthesized from 0.886 g (2.51 mmol) of compound (R1c) in the same manner as in the synthesis of compound (R2a).

Example 1 a) Nt-butoxycarbonyl-L-phenylalanyl-L-tryptophan benzyl ester (1a) 5.31 g (20.0 mmol) of Nt-butoxycarbonyl-L-phenylalanine 6.81 g (20.6 mmol) of L-tryptophan benzyl ester hydrochloride and 2.70 g of 1-hydroxybenzotriazole
(20.0 mmol) in dry tetrahydrofuran (T
HF) (50 ml) and stirred at -15 ° C for 15 minutes. To this suspension was added 1-ethyl-3- (3′-dimethylaminopropyl) carbodiimide hydrochloride (WSC.
4.17 g (21.75 mmol) of HCl) and 1.6 ml (22 mmol) of triethylamine were added, and the mixture was stirred at room temperature overnight. The solvent was distilled off, and the residue was dissolved in ethyl acetate. The organic layer was washed with 1N-HCl, water and saturated aqueous sodium bicarbonate in that order, dried over anhydrous sodium sulfate and concentrated, and the obtained residue was purified by column chromatography to give 8.15 g of the title compound (yield; 75.3). %).

1 H NMR (CDCl ₃ ) δ; 1.3
5 (9H, s), 2.99 (2H, d, J =
6 Hz), 3.15-3.35 (2H, m), 4.25
-4.95 (2H, m), 5.00-5.15 (2H,
m), 6.25-6.67 (2H, m), 7.00-
7.36 (15H, m), 7.85-8.00 (1H,
m)

B) L-Phenylalanyl-L-tryptophan benzyl ester hydrochloride (1b) 8.00 g (14.8 mmol) of compound (1a) was added to 90
% Formic acid (133 ml) and stirred at room temperature for 3 hours. The reaction solution was concentrated, and 1.4N hydrochloric acid-dioxane (1
2.5 ml) and stirred for 15 minutes. Ether (120 ml) was added to this solution, and the precipitated crystals were collected by filtration, washed with ether, and dried under reduced pressure at room temperature to obtain 6.24 g (yield: 88.3%) of the title compound.

1 H NMR (CD ₃ OD) δ; 2.8
8-3.35 (4H, m), 4.03-4.88 (2
H, m), 5.07 (2H, s), 7.00
-7.54 (15H, m)

C) N- (O-benzyl-P-2-phenethylphosphonyl) -L-phenylalanyl-L-tryptophan benzyl ester (1c) 956 mg (2.0 mmol) of compound (1b) was dried in methylene chloride. (8 ml), and triethylamine (0.84 ml, 6.0 mmol) was added thereto under cooling. Under ice-cooling and stirring, a solution (2.8 mmol) of the compound (R2b) in dry methylene chloride (5 ml) was added dropwise in about 15 minutes, and the mixture was stirred overnight. The reaction solution was washed with water, and the solvent was distilled off. The residue was dissolved in ethyl acetate, 1N hydrochloric acid, water, saturated aqueous sodium bicarbonate,
Washed with water and saturated saline in this order. The organic layer was dried over anhydrous sodium sulfate, and the solvent was distilled off. The residue was purified by silica gel column chromatography to give the title compound (411 m).
g (yield; 29.4%).

1H NMR (CDCl ₃ ) δ; 1.2
−2.0 (2H, m), 2.3-3.4 (6H,
m), 3.8-4.2 (6H, m), 6.4-8.0
(28H, m)

D) N- (2-phenethylphosphonyl)
-L-phenylalanyl-L-tryptophan disodium salt (1d) 82 mg (0.12 mmol) of compound (1c), ethanol (6.5 ml), water (6.5 ml), 19.5% sodium bicarbonate
mg (0.23 mmol) and a mixture of 13.5 mg of 10% palladium-carbon were stirred overnight at room temperature under a hydrogen atmosphere. After the catalyst was filtered off, the solvent was distilled off. The residue was dissolved in water and freeze-dried to obtain 71 mg (quantitative) of the title compound.

1H NMR (D ₂ O) δ: 1.1-
1.3 (2H, m), 2.2-2.4 (2H, m),
2.5-2.9 (2H, m), 3.1-3.3 (2
H, m), 3.8-4.0 (1H, m), 4.4-
4.6 (1H, m), 7.0-7.4 (15H, m)

Example 2 a) Nt-butoxycarbonyl-L-leucyl-L
-Tryptophan benzyl ester (2a) Nt-butoxycarbonyl-L-leucine 2.49 g
(10.0 mmol) and 3.31 g (10.0 mmol) of L-tryptophan benzyl ester hydrochloride,
Synthesized in the same manner as in the synthesis of compound (1a), 4.77 g (yield; 94%) of the title compound was obtained.

1 H NMR (CDCl ₃ ) δ; 0.8
5 (6H, d, J = 6 Hz), 1.0-1.8 (3H,
m), 1.40 (9H, s), 3.30 (2H, d, J
= 5 Hz), 3.8-4.2 (1H, m), 4.
5-5.2 (2H, m), 5.05 (2H, s),
6.48 (1H, d, J = 8 Hz), 6.7-
8.2 (11H, m)

B) L-leucyl-L-tryptophan benzyl ester hydrochloride (2b) Compound (2a) was synthesized from 4.56 g (9.0 mmol) of compound (2a) in the same manner as in the synthesis of compound (1b) to give the title compound. 3.89 g (yield; 97%) was obtained.

1 H NMR (DMSO-d ₆ ) δ;
0.6-2.0 (9H, m), 2.8-3.5 (2
H, m), 3.6-4.0 (1H, m), 4.4-
4.8 (1H, m), 5.02 (2H, s),
6.6-7.7 (10H, m), 8.28 (3H,
s), 9.18 (1H, d, J = 8 Hz), 1
0.99 (1H, s)

C) N- (O-benzyl-P-benzylphosphonyl) -L-leucyl-L-tryptophan benzyl ester (2c) 310 mg (0.7 mmol) of compound (2b) and 1.0 mg of compound (R2c) From the compound (1c) in the same manner as in the synthesis of the compound (1c).
4%).

1H NMR (CDCl ₃ ) δ; 0.7
3 (6H, d, J = 5 Hz), 1.0-1.7 (3H,
m), 2.6-3.4 (4H, m), 3.0-
4.0 (1H, m), 4.4-5.1 (5H, m),
6.6-8.5 (23H, m),

D) N-benzylphosphonyl-L-leucyl-L-tryptophan disodium salt (2d) From 62 mg (0.1 mmol) of compound (2c), it was synthesized in the same manner as in the synthesis of compound (1d). , Title compound 4
3 mg (yield; 88%) were obtained.

1H NMR (D ₂ O) δ; 0.74
(6H, t, J = 5 Hz), 0.8-1.6 (3H,
m), 2.74 (2H, d, J = 19 Hz), 2.9-
3.7 (3H, m), 4.3-4.6 (1H, m),
6.8-7.9 (10H, m)

Example 3 a) N- (O-benzyl-P-2-phenethylphosphonyl) -L-leucyl-L-tryptophan benzyl ester (3a) 310 mg (0.7 mmol) of compound (2b) and Compound (R2b) was synthesized from 1.0 mmol by the same method as in the synthesis of compound (1c) to give 70 mg of the title compound (yield;
15%).

1H NMR (CDCl ₃ ) δ; 0.8
1 (6H, d, J = 5 Hz), 1.0-2.2 (5H,
m), 2.5-3.0 (2H, m), 3.0-
4.0 (3H, m), 4.4-5.1 (5H, m),
6.5-7.7 (22H, m), 8.32 (1H,
s)

B) N- (2-phenethylphosphonyl)
-L-leucyl-L-tryptophan disodium salt (3b) A compound (3a) was synthesized from 74 mg (0.11 mmol) in the same manner as in the synthesis of compound (1d), and 55 mg of the title compound (yield; 95%) ) Got.

1H NMR (D ₂ O) δ; 0.4-
1.0 (6H, m), 1.0-2.0 (5H, m),
2.4-2.9 (2H, m), 2.9-3.7 (3H,
m), 4.3-4.6 (1H, m), 6.8-7.9
(10H, m)

Example 4 a) Nt-butoxycarbonyl-L-isoleucyl-L-tryptophan benzyl ester (4a) Nt-butoxycarbonyl-L-isoleucine 2.3
Compound (1a) was synthesized from 1 g (10 mmol) and 3.31 g (10 mmol) of L-tryptophan benzyl ester hydrochloride in the same manner as in the synthesis of compound (1a) to obtain 3.58 g (yield; 70%) of the title compound. Was.

1 H NMR (CDCl ₃ ) δ; 0.8
3 (6H, d, J = 7 Hz), 1.0-1.9 (3H,
m), 1.42 (9H, s), 3.29 (2H, d, J
= 5 Hz), 3.7-4.0 (1H, m), 4.7
−5.2 (2H, m), 5.04 (2H, s),
6.30 (1H, d, J = 8 Hz), 6.7-8.
2 (11H, m)

B) L-isoleucyl-L-tryptophan benzyl ester hydrochloride (4b) The compound (4a) was synthesized from 3.30 g (6.5 mmol) of compound (4a) in the same manner as in the synthesis of compound (1b) to give the title compound 2.55 g (yield; 88%) was obtained.

1 H NMR (DMSO-d ₆ ) δ;
0.6-2.0 (9H, m), 2.6-3.8 (3
H, m), 4.3-4.7 (1H, m), 5.00
(2H, s), 6.6-7.6 (10H, m), 8.
27 (3H, s), 9.07 (1H, d, J = 8H
z), 10.99 (1H, s)

C) N- (O-benzyl-P-2-phenethylphosphonyl) -L-isoleucyl-L-tryptophan benzyl ester (4c) 400 mg (1.0 mmol) of compound (4b) and compound (R2b) 1 Of the title compound (yield; 98 mg) in a similar manner to that of compound (1c).
21%).

1H NMR (CDCl ₃ ) δ; 0.5
-2.2 (11H, m), 2.5-3.8 (5H,
m), 4.5-5.1 (1H, m), 5.01 (2
H, s), 5.04 (2H, s) 6.3.
7.6 (22H, m), 7.8-8.1 (1H, m)

D) N- (2-phenethylphosphonyl)
-L-isoleucyl-L-tryptophan disodium salt (4d) Compound (4c) was synthesized from 98 mg (0.15 mmol) of compound (4c) in the same manner as in the synthesis of compound (1d), to give 76 mg of the title compound (yield; 96%). ) Got.

1H NMR (D ₂ O) δ; 0.3-
1.0 (8H, m), 1.0-2.0 (3H, m),
2.4-2.9 (2H, m), 2.9-3.8 (3
H, m), 4.3-4.7 (1H, m), 6.8-
7.8 (10H, m)

Example 5 a) N-benzyloxycarbonyl-L-isoleucyl-L-tryptophan methyl ester (5a) N-benzyloxycarbonyl-L-isoleucine
30 g (20.0 mmol) and 5.09 g (20.0 mmol) of L-tryptophan methyl ester hydrochloride were synthesized in the same manner as for compound (1a) to give 7.14 g of the title compound (yield: 75.9). %).

1 H NMR (CDCl ₃ ) δ; 0.8
2-0.89 (6H, m), 1.04-1.64 (2
H, m), 1.79-1.80 (1H, m), 3.2
9-3.34 (2H, m), 3.66 (3H, s),
4.04-4.06 (1H, m), 4.89-4.93
(1H, m), 5.05 (2H, s), 5.34 (1
H, brs), 6.43 (1H, m), 6.9
3-8.05 (11H, m)

B) L-isoleucyl-L-tryptophan methyl ester (5b) 7.14 g (15.2 mmol) of compound (5a) was dissolved in methanol (50 ml), and 10% palladium carbon (710 mg) was added thereto. Stirred under a hydrogen atmosphere overnight. The catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure to give 3.06 g of the title compound (yield; 63.5%).
I got

1 H NMR (CD ₃ OD) δ; 0.8
0-1.68 (9H, m), 3.11-3.33 (3
H, m), 3.66 (3H, s), 4.7
4-4.85 (1H, m), 6.97-7.62 (6
H, m)

C) N- (O-benzyl-P-2-phenethylphosphonyl) -L-isoleucyl-L-tryptophan methyl ester (5c) 663 mg (2.0 mmol) of compound (5b) and compound (R2b) 2 From 2.8 mmol, the title compound was synthesized in the same manner as in the synthesis of compound (1c) to obtain 258 mg (yield; 21.9%) of the title compound.

1H NMR (CDCl ₃ ) δ; 0.6
-2.2 (11H, m), 2.6-3.0 (2H,
m), 3.2-3.35 (2H, m), 3.4-3.
8 (4H, m), 4.5-5.1 (3H, m),
7.4-8.2 (18H, m)

D) N- (2-phenethylphosphonyl)
-L-isoleucyl-L-tryptophan methyl ester sodium salt << (1S, 2S) -1-[(s)-
2- (3-Indolyl) -1-methoxycarbonylethylcarbamoyl] -2-methylbutylaminophenylethylphosphinic acid sodium salt> (5d) From compound (5c) 136 mg (0.23 mmol),
Synthesized in the same manner as in the synthesis of compound (1d), 114 mg (yield; 95.0%) of the title compound was obtained.

1H NMR (D ₂ O) δ; 0.7-
1.3 (8H, m), 1.4-1.8 (3H, m),
2.5-2.7 (2H, m), 3.2-3.4 (2H,
m), 3.4-3.6 (4H, m), 4.6-4.8
(1H, m), 7.0-7.6 (10H, m)

Example 6 a) N- (O-benzyl-P-4-phenylbutylphosphonyl) -L-leucyl-L-tryptophan benzyl ester (6a) 2350 mg (5.3 mmol) of compound (2b) The compound (R2a) was synthesized from 7.5 mmol in the same manner as in the synthesis of the compound (1c) to obtain 463 mg (yield; 13.7%) of the title compound.

1H NMR (CDCl ₃ ) δ; 0.6
−1.0 (6H, m), 1.2−2.1 (9H,
m), 2.4-2.6 (2H, m), 3.15-
3.35 (2H, m), 3.6-4.0 (1H, m),
4.65-5.25 (5H, m), 6.8-7.6
(22H, m), 8.1-8.4 (1H, m)

B) N- (4-phenylbutylphosphonyl) -L-leucyl-L-tryptophan disodium salt (6b) From compound (6a) 232 mg (0.35 mmol),
Synthesized in the same manner as in the synthesis of compound (1d), 181 mg (yield; 92.7%) of the title compound was obtained.

1H NMR (CD ₃ OD) δ; 0.8
1-0.90 (6H, m), 1.47-1.66 (9
H, m), 2.51-2.61 (2H, m), 3.1
7-3.58 (3H, m), 4.56-4.59 (1
H, m), 6.93-7.63 (10H, m)

Example 7 a) N- (O-benzyl-P-4-phenylbutylphosphonyl) -L-isoleucyl-L-tryptophan
Benzyl ester (7a) The compound (4c) was synthesized from 2110 mg (4.8 mmol) of the compound (4b) and 6.8 mmol of the compound (R2a) in the same manner as in the synthesis of the compound (1c).
(Yield; 30.3%).

1H NMR (CDCl ₃ ) δ; 0.6
−0.8 (6H, m), 0.8-1.75 (9
H, m), 2.3-2.55 (2H, m), 3.0
-3.3 (2H, m), 3.45-3.7 (1H,
m), 4.55-5.05 (5H, m), 6.7-
7.55 (22H, m), 8.1-8.4 (1H,
m)

B) N- (4-phenylbutylphosphonyl) -L-isoleucyl-L-tryptophan disodium salt (7b) From compound (7a) (232 mg, 0.35 mmol),
Synthesized in the same manner as in the synthesis of compound (1d), 209 mg (yield; 99.0%) of the title compound was obtained.

1H NMR (CD ₃ OD) δ; 0.6
3-1.25 (8H, m), 1.44-1.67 (7
H, m), 2.52-2.61 (2H, m), 3.1
4-3.53 (3H, m), 4.61-4.63 (1
H, m), 6.94-7.63 (10H, m)

Comparative Example 1 a) Nt-butoxycarbonyl-L-alanyl-L
-Tryptophan benzyl ester (1a ratio) 3.78 g of Nt-butoxycarbonyl-L-alanine
(20.0 mmol) and 6.81 g (20.6 mmol) of L-tryptophan benzyl ester hydrochloride,
The title compound 1 was synthesized in the same manner as in the compound (1a).
0.74 g (quantitative) was obtained.

1 H NMR (CDCl ₃ ) δ; 1.2
7 (3H, d, J = 7 Hz), 1.40 (9H,
s), 3.31 (2H, d, J = 5 Hz), 4.1
(1H, brs), 4.92-4.96 (1H, m),
5.07 (2H, s), 6.55 (1H, d, J =
8Hz), 6.81 (1H, s), 7.07-7.5
1 (10H, m), 8.08 (1H, brs)

B) L-alanyl-L-tryptophan benzyl ester hydrochloride (comparative 1b) From compound (comparative 1a) 10.8 g (20.0 mmol), compound (1b) was synthesized in the same manner as compound (1b). 6.29 g (yield; 78.3%) of the title compound was obtained.

1H NMR (DMSO-d ₆ ) δ;
1.34 (3H, d, J = 6 Hz), 3.11-3.2
6 (2H, m), 3.87-3.88 (1H, m),
4.59-4.65 (1H, m), 6.97-7.51
(10H, m), 8.31 (3H, brs), 9.10
(1H, d, J = 7 Hz), 11.03 (1H, s)

C) N- (O-benzyl-P-2-phenethylphosphonyl) -L-alanyl-L-tryptophan benzyl ester (1c ratio) 804 mg (2.0 mmol) of compound (1b) and compound (R2b) ) The compound (1c) was synthesized from 2.8 mmol in the same manner as in the synthesis of the compound (1c) to obtain 288 mg (yield; 23.1%) of the title compound.

1H NMR (CDCl ₃ ) δ; 1.2
4 (3H, dd, J = 7, 16 Hz), 1.8-2.0
(2H, m), 2.7-2.9 (2H, m), 3.2
-3.4 (2H, m), 3.7-3.9 (1H,
m), 4.7-5.2 (5H, m), 6.7-7.9
(23H, m)

D) N- (2-phenethylphosphonyl)
-L-alanyl-L-tryptophan disodium salt (ratio 1d) From compound (ratio 1c) 146 mg (0.23 mmol), synthesized in the same manner as in the synthesis of compound (1d), 117 mg of the title compound (quantitative) I got

1H NMR (D ₂ O) δ; 1.14
(3H, d, J = 7 Hz), 1.6-1.8 (2H,
m), 2.5-2.7 (2H, m), 3.1-
3.4 (2H, m), 3.6-3.7 (1H, m),
4.4-4.6 (1H, m), 7.1-7.7 (1
0H, m)

[0080]

Industrial Applicability The compound of the present invention has a strong inhibitory activity on endothelin converting enzyme, has an excellent blood pressure lowering action, and has an active ingredient (endothelin altering agent) for a medicine against various vascular diseases.
It is a substance useful as an enzyme inhibitor .

──────────────────────────────────────────────────続 き Continuation of the front page (51) Int.Cl. ⁶ identification code FI // C07K 101: 02 (58) Investigated field (Int.Cl. ⁶ , DB name) C07K 5/062 A61K 31/66 ABN A61K 31/66 ABU A61K 31/66 ACV C07K 5/065 C07K 101: 02 CA (STN) REGISTRY (STN) WPI (DIALOG)

Claims (2)

(57) [Claims] [Claim 1] The following formula (1) (Wherein A represents an amino acid residue selected from the group consisting of leucine, isoleucine and phenylalanine;
rp represents a tryptophan residue, R may be the same or different and represents a lower alkyl group, a lower alkoxy group, or a halogen atom, X represents a hydrogen atom or an alkali metal, Y represents a hydrogen atom, an alkali metal Or a lower alkyl group, m is an integer of 0 to 3, and n is an integer of 1 to 4). 2. A phosphonic acid represented by the above formula (1)
An endothelin converting enzyme inhibitor comprising a amide derivative.