JPS6137790A - Phosphorus-containing peptide derivative - Google Patents

Abstract

NEW MATERIAL:A compound shown by the formula I {R<1> is lower alkyl; X is COOH, hydroxymethyl, -CO2R<2>[R<2> is lower alkyl, (substituted) aryl, or aralkyl], -CH2OR<2>, or -CH2OCOR<3>[R<3> is H, lower alkyl, (substituted)aryl, or aralkyl)}, and its salt. EXAMPLE:(1R)-2-( 4-Hydroxyphenyl )-1-{N-[(1S,2S)-methoxycarbonyl-2-methylbutyl] carbamoyl-L-tyrosyl}aminoethylphosphonic acid. USE:An inhibitor against an enzyme capable of converting angiotensin, or an antihypertensive. PREPARATION:A compound shown by the formula II(X' is X or protected COOH or amino) is condensed with a compound shown by the formula III (Y is phenolic OH-protecting group; Z is lower alkyl) to give a compound shown by the formula IV, and the protecting groups Y and Z are eliminated.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a novel phospho-containing peptide having angiotensin-converting enzyme inhibiting action and, in turn, antihypertensive action, and salts thereof.

BACKGROUND ART Until now, as phosphopeptide-containing compounds having angiotensin-converting enzyme inhibitory action, the compounds represented by the following formula and derivatives thereof are typically known: CB, D, Thorsett et al. , P
roc, Natl, Acad.

Sci, IJs 8, 79. 2176 (19
82) 3.

Furthermore, there is a series of applications filed by Honde Kōnin regarding phosphopeptides that have strong angiotensin-converting enzyme inhibitory effects and antihypertensive effects (Japanese Patent Laid-Open No. 57-156498).
．． 58-31992. 58-85896.
58-85897゜59-51294. Same 59
-51295).

Problems to be Solved by the Invention Based on these findings, we further investigated the synthesis of novel phosphopeptides, and found that the general formula [1
) has been found to exhibit excellent angiotensin-converting enzyme inhibitory activity and antihypertensive activity.
The present invention has now been completed.

Means for Solving the Problems The present invention provides compounds of the formula CI] [wherein R' is lower alkyl]. X is a carboxyl group, a hydroxymethyl group, -CDJ" (in the formula, R2
is lower alkyl, unsubstituted or substituted aryl, or aralkyl), -Ct120R2 (wherein R2
is as defined above) or -Cl (20COR3 (wherein R3 is hydrogen, lower alkyl, unsubstituted or substituted aryl, or aralkyl)) (hereinafter referred to as (referred to as compound [I]. The same applies to compounds with other formula numbers) and its salts.

Next, the present invention will be explained in more detail.

First, the range of compounds included in formula CI] is clearly specified. In the definition of R1 in formula [I], lower alkyl means a straight chain or branched alkyl having 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl.

1-propyl, 1-methylpropyl, 2-methylpropyl, n-butyl, l-butyl, t-butyl.

, n-pentyl, neopentyl, hexyl and the like. Preferably l-propyl, 1-methylpropyl, 2
-Methylpropyl, etc.

In the definition of R2 and Ro, lower alkyl means a straight chain or branched alkyl having 1 to 6 carbon atoms, and the same examples as above are exemplified, but preferred examples include methyl, ethyl, propyl, etc. . In the definition of R2 and R3, aryl is phenyl, naphthyl, biphenyl, etc., alkyl having 1 to 3 carbon atoms, hydroxy, alkoxy having 1 to 3 carbon atoms, and halogen (chlorine, bromine, etc.).

It may be substituted with nitro, carboxyl, amino group, etc. In the definition of R1, aralkyl means that the aryl part is buenyl, naphthyl, biphenyl, etc., and the alkyl part has 1 to 3 carbon atoms.

Compound [I] can form a base addition salt.

Such salts include ammonium salts.

Alkali metal salts such as lithium, sodium and potassium salts; alkaline earth metal salts such as calcium and magnesium salts; salts with organic bases such as triethylamine, morpholine, piperidine and dicyclohexylamine; and with amino acids such as arginine and lysine. Contains salt, etc.

Although non-toxic pharmaceutically acceptable salts are preferred, other salts are also useful in isolating and purifying the product.

These salts can be made by conventional methods. For example, compound [I] is reacted with one or more equivalents of a suitable base or acid relative to compound [I] in a solvent such as water or an alcohol, and the solvent is removed by vacuum or freeze-drying; Compound [I
] can also be formed by replacing the cation of the salt with another cation.

In the formula [I, I, the carbon atoms bonded to R1 and -cH2 are asymmetric carbon atoms. According to the invention,
By selecting the starting materials and intermediates used, racemic,
Enantiomeric and diastereomeric products can be obtained. In addition, when diastereomeric mixtures are obtained, they can be separated by conventional chromatography or fractional crystallization methods. In the present invention, the asymmetric carbon configuration has pharmacological activity in both R and S configurations, but it is more preferable that in the formula [N, the phosphorus atom is A compound is selected in which the carbon to which R' is bonded has the R-configuration, the carbon not bonded to the other phosphorus atom has the S-configuration, and the carbon to which R' is bonded has the S-configuration.

Next, a method for producing compound CI) will be explained.

Compound [I] is synthesized by the following steps.

(In each of the above formulas, Y is a protecting group for a phenolic hydroxyl group.

Z is lower alkyl. X' has the same meaning as X, or when X contains a carboxyl group or an amino group, it means an X in which these groups are protected. ) [Step-1] Compound [III], which is the starting compound of the present invention, can be synthesized from the compound described in JP-A-59-51294 by the present applicant and 4-nitrophenyl chloroformate ( (See Reference Example 1).

The condensation of compound [11) and CI[] is carried out by the condensation of compound [■
] and [II[] to proceed. Ethyl acetate and tetrahydrofuran are used as solvents.

Dioxane, chloroform, dichloromethane.

Acetone, N,N-dimethylformamide, pyridine, etc. may be used alone or in combination.

The reaction is usually carried out at 0°C to room temperature and is completed in 1 to 15 hours.

When a carboxyl group and an amino group are present in X of compound [II), it is desirable to carry out the condensation reaction while these groups are protected. For example, in the case of a carboxyl group, it can be condensed with a methyl ester and then deprotected by alkaline hydrolysis. In the case of an amino group, it can be condensed while protected with a t-butoxycarbonyl group and selectively deprotected by acid treatment such as hydrogen chloride/ethyl acetate.

[Step-2] Compound (IF is a compound in which Y is benzyl, trityl, benzyloxycarbonyl, t-butyl, t-butoxycarbonyl, and Z is lower alkyl such as methyl or ethyl) [
IV] by treatment with a solution of hydrogen bromide in acetic acid or trifluoroacetic acid. The reaction is usually completed in 3 to 15 hours at room temperature.

Next, examples, reference examples, and experimental examples of the present invention will be shown.

Example 1 (IR)-2-(4-benzyloxyfer)-1-[
N-(4-nitrophenyloxycarbonyl)-〇-benzyl-monotyrosyl]aminoethylphosphonic acid diethyl ester (m a ) 2345 mg (3 mmol)
A solution of 1,742 mg (12 m mo+) of L-isoleucine methyl ester in ethyl acetate (<10 ml) was added dropwise to a suspension of 1,742 mg (12 m mo+) in ethyl acetate (20 ml), and the mixture was stirred overnight at room temperature. Add 30ml of ethyl acetate to the homogeneous reaction solution,
This solution was washed with an IN sodium hydroxide solution until the yellow color of 4-ditrophenol disappeared, and then washed with a 5% aqueous citric acid solution and a saturated saline solution. After drying over anhydrous sulfuric acid (IJ), the solvent was removed under reduced pressure to give a glass-like rVa
2009 mg (85%) was obtained.

Compound rVa552■, anisole 0.5ml 25%
It was dissolved in 50+1 hydrogen bromide/acetic acid solution and stirred at room temperature for 6 hours. Volatile substances were removed under reduced pressure, and the residue was triturated with ethyl ether to obtain a gummy substance. This compound was reprecipitated from methanol-chloroform-ethyl ether and dried under reduced pressure to obtain a pale yellow powder.

A solution of this powder in methanol (2 ml) was heated under reflux for about 10 minutes, the solvent was removed, and the residue was lyophilized from water to obtain 219 mg (57%) of Ia in the form of a pale yellow powder.

'H-NMR (C port, 00-CDCj!3)
6 7.2-6.5 (m, 8H).

3.67 (s, 3) 1), 3J -2,35 (m, 4
H), 2.0-1.0 (m, 3) 1), i, 0-
0.6 (m, 6H).

[α] = −54,8” (c =0.50.
Methanol) Example 2 A solution of 945 ml (1.2 mmol) of compound IVa in methanol (15 ml) was ice-cooled, and 1.8 ml (1.8 mmol) of IN sodium hydroxide aqueous solution was added thereto for 3 hours and then overnight at room temperature. Stirred. 5% to reaction mixture
The mixture was acidified with 20 ml of citric acid aqueous solution and extracted with ethyl acetate (2×40 ml). The ethyl acetate layer was washed with water, dried over IJ sulfate, and then the solvent was removed under reduced pressure to obtain 857 mg of a pale yellow glassy substance.

This compound was treated in the same manner as in Example 1(ii) to obtain 1b in the form of a white powder.

'H-NMR (C port 300 + CDCR6)
δ 7.3-6.4 (m, 8H).

3.4-3.2 (m, 4H), 2.0-1.0 (m,
3゛H), 1.0-0.4 (m, 6H) [α] = -69.3° (c ~0.49. Methanol) Example 3 Xyphenyl)ethylphosphonic acid He) Same as Example 1 By the method described above, white powder Ic was obtained from L-inleucinol and compound 111a.

'l(-NMR(C[1301) + C[1CL
) δ 7.2-6.5 (m, 8) 1).

3.8-2.4 (n+, 7H), 1.7-1.0 (+
n, ]), 1.0 to 0.6 (m, 6) 1) [α] = -71.5″' (c = 0.40. Methanol) Reference Example 1 (IRII-(○-Benzylene) tyrosyl)amino-
2-(4-benzyloxyphenyl)ethylphosphonic acid diethyl ester hydrochloride 5225mg (8m mo
2177 mg of 4-nitrophenyl chloroformate (10.8 mm
ol), pyridine 1.52 ml (18,8 m m
ol) was added and stirred at room temperature for 4.5 hours. To the reaction solution were added 70ml of ethyl acetate and 20++1 ml of 1.5% citric acid aqueous solution for two-phase partitioning, and the organic layer was collected. This was washed with a 5% aqueous citric acid solution and then with saturated saline (2×20 ml each).

It was dried with anhydrous sodium sulfate.

The solvent was removed under reduced pressure, 20 ml of ethyl ether was added, and when the mixture was left to stand, white crystals precipitated. The crystals were separated by filtration, washed with ethyl ether, dried under reduced pressure, and
5620 mg (90%) was obtained.

Melting point 119; 5-120.5°C 'II-NMR (CDCji!3) δ 8.09 (d,
2H), 7.5-6.75 (m, 20tl), 4
．． 97 +4.94 (2s, 4fl), 4.9~
4.3 (m.

2H), 4.07 (m, 4tl), 3.4-2.6
(m, 48), 1.31 +1, 23 (2t, 6
H, J=7Hz) [α] −-51,8″(c ~0
．． 50. Chloroform) Experimental Example The angiotensin converting enzyme inhibitory activity of the compounds obtained according to the present invention was determined by the method of D., W. Cushman et al., CBiochem, Pharmaco 1. , 20.1
63T (1!171)). That is,
Synthetic substrate N by rabbit lung angiotensin converting enzyme
The inhibitory effect of the test compound on the hydrolysis process of -benzoyl-glycyl-histidyl-histidyl-leucine was measured. An aqueous solution containing a certain amount of enzyme, substrate, and test compound was shaken at 37° C. for 30 minutes, and the amount of N-benzoyl-glycine produced was quantified by absorbance measurement (228 nm).

The concentration of the test compound was varied, and the compound concentration (ICso) that inhibited the enzyme activity by 50% was determined.

The results are shown in Table 1.

Table 1 Compounds with angiotensin-converting enzyme inhibitory activity of Compound I [
I] and its salts have angiotensin-converting enzyme inhibitory action and, in turn, antihypertensive action.

Patent applicant (102) Kyowa Hakko Kogyo Co., Ltd. 212/

Claims (5)

[Claims] (1) Formula [I] ▲There are mathematical formulas, chemical formulas, tables, etc.▼[I] [In the formula, R^1 is lower alkyl. X is a carboxyl group, a hydroxymethyl group, -CO_2R^2 (in the formula,
R^2 is lower alkyl, unsubstituted or substituted aryl, or aralkyl), -CH_2OR^2 (wherein R^2 is as defined above) or -CH_2OC
OR^3, where R^3 is hydrogen, lower alkyl, unsubstituted or substituted aryl, or aralkyl. ] A phosphopeptide-containing derivative and a salt thereof. (2) The phosphopeptide-containing derivative and its salt according to claim 1, wherein in formula [I], R^1 is 1-methylpropyl. (3) The phosphopeptide-containing derivative and its salt according to claim 1, wherein in formula [I], X is a carboxyl group, a hydroxymethyl group, or a lower alkoxycarbonyl group. (4) The phosphopeptide-containing derivative and its salt according to claim 1, wherein in formula [I], the carbon to which R^1 and ▲ are represented by mathematical formulas, chemical formulas, tables, etc. are optically active. (5) In formula [I], there are mathematical formulas, chemical formulas, tables, etc. Of the two carbons to which ▼ is bonded, the carbon that is also bonded to the phosphorus atom has an R-configuration, and the other phosphorus atom has an R-configuration. The carbon not bonded to the S-configuration, and also the R^1
5. The phosphopeptide derivative and its salt according to claim 4, wherein the carbon bonded to is in S-configuration.