JPS624286A - Novel amino acid derivative - Google Patents

Abstract

NEW MATERIAL:A compound expressed by formula I (X is lower alkoxycarbonyl or hydroxymethyl; Y is H or OH; His is L-histidyl; n is 0 or 1; Z is -O- or -NH-; R is 1-7C alkyl) and acid addition salt thereof. EXAMPLE:Isopropyl (2RS, 3S)-3-{N-[2-(1-naphthylmethyl)-3-(2S-methoxycarbonyl pyrrolid inocarbonyl)propionyl]-L-histidyl}amino-2-hydroxy-5-methylh-exanoa te. USE:A remedy for hypertension, having inhibitory action on human renin and orally administrable. PREPARATION:A compound expressed by formula II is condensed with a compound expressed by formula III [C(L) is L-configuration; C* is S configuration] using diphenylphosphoric acid azide.

Description

Detailed Description of the Invention (1. Title of the Invention [Industrial Application Field] The object of the present invention is to obtain a compound having a general formula (wherein X is a lower alkoxycarbonyl group or a hydroxymethyl group, and Y is a lower alkoxycarbonyl group or a hydroxymethyl group) A hydrogen atom or a hydroxyl group, His is L
- histidyl group, n is 0 or 1, Z is 10- or -NH-, R is a straight or branched alkyl group having 1 to 7 carbon atoms) An object of the present invention is to provide novel amino acid derivatives and their pharmacologically acceptable acid addition salts. In more detail,
To provide a novel amino acid derivative represented by the above general formula and a pharmacologically acceptable acid addition salt thereof, which has a human renin inhibitory effect and is useful as an orally administrable therapeutic agent for hypertension. It is.

[Conventional technology]

Renin is a proteolytic enzyme released from paraglobular cells of the kidney. This substance reacts with the renin substrate present in the α2 globulin fraction of plasma, and angiotensin I (ang
iotensin I). The generated angiotensin I is converted to angiotensin II by angiotensin converting enzyme.
) is converted to This angiotensin ■ has a vasoconstrictive effect, and also acts on the adrenal cortex, and induces aldosterone (aldo-sterone), which affects sodium and water metabolism.
One of the factors that causes hypertension.

Compounds that inhibit the reaction between renin and renin substrates and suppress the production of angiotensin 2 are attracting attention as antihypertensive agents with a new mechanism of action, and their development is strongly desired.

Until now, the reaction between lane and renin substrate has been inhibited, i.e.
Many peptide derivatives are known as compounds that inhibit renin activity (Japanese Patent Publication No. 1983-
No. 39149, Japanese Patent Publication No. 59-110661
No., 1982-155345, 1984-2278
No. 51, European Patent Publication No. 707029, No. 7
(No. 7028, No. 81783) Among these patent applications, especially Japanese Patent Publication No. 1983
-155345 has the general formula (A in the formula is a hydrogen atom, a phenyl group, a 10.11-dihydro-5-dibenzo[a, d] cycloheptenyl group, and B is one rho, -CH=CH -1-C)+
2-, etc., and p and q
may be the same or different, each being 0 or 1 to 3
is an integer, D is a hydrogen atom, a lower alkyl group, a lower alkenyl group, a phenyl group, a phenylalkyl group, etc., E is a phenyl group, a cyclohexyl group, an isopropyl group, etc., and His is an L-histidyl group. , r is an amino acid residue, for example, L-leucyl group, L-isoleucyl group, L-leucyl-phenylalanyl group, L-
phenylalanyl-L-phenylalanyl group, L-alanyl-L-phenylalanyl group, etc., and G is a protecting group at the C-terminus of the amino acid, such as an amino group, an alkylamino group, an allylalkylamino group, an alkoxy group, etc. )
Discloses a peptide derivative represented by:

Also, in Japanese Patent Publication No. 59-227851,
General formula (where R'CD- is an acyl group of an N-substituted amino acid, such as benzyloxycarbonyl group, t-butoxycarbonyl group, α-naphthoxyacetyl group, phenylphthyryl group, 3-(3-nitro- Amino acids whose amine groups are substituted with 2-pyridinesulfenyl)thiopropionyl groups, 9-fluorenylmethyloxycarbonyl groups, etc., such as phenylalanyl groups, L-triftphyll groups,
Tyrosyl group, L-cystyl group, 3-(1-naphthyl)-
-alanyl group, L-phenylglycyl group, ■-amino-4-phenylbutyryl group, 1,2゜3.4-tetrahydro-β-carboline-5-L-carbo group, or formyl group, and R3 is Discloses a peptide derivative represented by a hydrogen atom, an alkyl group, or a substituted alkyl group having a hydroxyl group, a mercapto group, an ami7 group, a carbamoyl group, a formyl group, an aryl group, or a heterocyclic group as a substituent. are doing.

[Problem that the invention seeks to solve]

Most of the compounds disclosed in the above-mentioned patent applications are polypeptides, which are difficult to synthesize, and are difficult to synthesize using proteolytic enzymes in the body, such as chymotrypsis 7 (chymotrypsis 7).
rypsin), and cannot be expected to exert its pharmacological effects upon oral administration.

The peptide derivative represented by the general formula (II) is a tri- or tetra-peptide derivative, and although it is relatively easy to manufacture compared to other polypeptide derivatives, this compound is also disclosed in the above-mentioned patent application etc. Like polypeptide derivatives, it is unstable to proteolytic enzymes, and it is difficult to expect that it will exhibit its medicinal efficacy when administered orally.

Furthermore, although the compound represented by formula (I[) is easier to produce than other peptide derivatives, this compound is also susceptible to proteolytic enzymes like the polypeptide derivatives disclosed in the aforementioned patent applications. On the other hand, it is unstable and it is difficult to expect that it will exhibit its medicinal efficacy when administered orally.

As a result of various studies to solve these problems, the present inventors have found that the amino acid derivatives represented by the general formula (I) and their pharmacologically acceptable acid addition salts can be produced relatively easily. It has been found that it has a strong renin activity inhibiting effect, has low toxicity, can be administered orally, and can solve the above-mentioned problems.

(2) Structure of the invention [Means and effects for solving the problems] The amino acid derivatives of the present invention represented by the general formula (I) and their pharmacologically acceptable acid addition salts are human renin-sheep renin. It exhibits a strong renin activity inhibitory effect in substrate systems and human high-renin plasma, and is stable against proteolytic enzymes such as chymotrypsin and pepsin. Additionally, this product exhibits a clear hypotensive effect when injected orally or intravenously in monkeys with high renin status.

This means that the amino acid derivatives of the present invention represented by the general formula (I) and their pharmacologically acceptable acid addition salts have a strong renin activity inhibiting effect, and can be administered orally with low toxicity. It has been shown to be useful as a therapeutic agent.

The amino acid derivative represented by the general formula (I) of the present invention is a compound represented by the general formula (in which X and Y have the same meanings as above), and the general formula has the same meanings as above. ) using diphenylphosphoric azide.

The general formula (rV
The carboxylic acid represented by ) can be produced by a method described in the literature or a method similar thereto. For example, 1-naphthaldehyde and diethyl succinate are reacted to obtain a compound represented by the formula, which is hydrolyzed with sodium hydroxide to form a dicarboxylic acid, and then ring-closed with acetic anhydride to obtain the compound represented by the formula. The succinic anhydride derivative shown is obtained. This succinic anhydride derivative of formula (■) is reacted with a cyclic amine represented by the general formula (X and Y in the formula have the same meanings as above). It can be produced by obtaining a compound represented by (having the same meaning as ) and hydrogenating this in the presence of palladium on carbon.

Furthermore, the compound represented by the above general formula (IV) in which X is a hydroxymethyl group, the compound represented by the above general formula (IV) in which X is a lower alkoxycarbonyl group is treated with a suitable reducing agent. , for example, by reduction using sodium borohydroxide.

The compound of the general formula (V) used as the other starting material in the production method of the present invention can be prepared by protecting the amine group with an appropriate protecting group and reacting histidine methyl ester with hydrazine in a methanol solution. ) has the same meaning as ).

After reacting this compound with isoamyl nitrite, the general formula (C, n, Z and R in the formula have the same meanings as above)
It can be produced by reacting with a compound represented by and then removing the protecting group.

The compound represented by the general formula (X I ) can be produced by a method described in the literature or a method similar thereto.

For example, a compound of general formula (XI) in which n is 0 is described in The Journal of Organic Chemistry [J, Org, 'C'4+em,] Vol. 45, 2.
3-amino-2-hydroxy- by a method similar to that described on pages 288-2290 ('1980).
It is produced by obtaining 5-methylhexanoic acid and esterifying or amidating it by a conventional method.

Further, the compound of general formula (XI) in which n is 1 can be obtained by esterifying statin or N-(tert-butyloxycarbonyl)statin by a conventional method. is produced by amidation.

The reaction between the compound represented by the general formula (rV) of the present invention and the compound represented by the general formula (V) can be carried out according to a conventional method. To carry out this reaction suitably, the general formula (I'
A compound represented by V) and an equimolar amount of the general formula (V
) and the compound represented by N.

Dissolved in N-dimethylformamide, stirred and cooled with water, and added equimolar amounts of diphenyl phosphoryl azide and the amino acid derivative represented by the general formula (I) of the present invention, including the asymmetric carbon of the histidine moiety. There are 6 asymmetric carbon atoms, and various isomers exist depending on the configuration of substituents at each asymmetric carbon 15.

The configuration of substituents at these asymmetric carbon atoms is expressed by the general formula (
Although it affects the renin inhibitory activity of the compound represented by I), the present invention does not particularly limit the configuration of the asymmetric carbon atoms other than the one-histidine moiety regarding these isomers.

In the amino acid derivative represented by the general formula (I) of the present invention, the configuration of the carbon atom substituted with the amino group in the compound moiety represented by the general formula (XI) is preferably the S configuration, The configuration of the carbon atom on which the hydroxyl group is substituted has a relatively small effect on the activity, and may be either the S configuration or the R configuration, or a mixture thereof.

The steric structure of the carboxylic acid represented by the general formula (IV) also affects the renin inhibitory activity of the compound represented by the general formula (I) of the present invention to some extent. In this acyl part, α−
Strong inhibitory activity is observed in either the S configuration or the R configuration of the carbon atom substituted with the naphthylmethyl group, but a mixture thereof may be used.

The optically active starting materials used in the production of such optically active compounds are produced by optical resolution by conventional methods or by using optically active compounds.

For example, a compound represented by the general formula (XI) in which the carbon atom substituted with the amino group has the S configuration is produced by using L-leucine or statin.

The compound represented by the general formula (I) of the present invention can be converted into a pharmacologically acceptable acid addition salt according to a conventional method,
Examples of these salts include hydrochloride, sulfonate, p-)reninsulfonate, acetate, and citrate. These acid addition salts also have a strong renin activity inhibiting effect, are stable to proteolytic enzymes, and clearly lower blood pressure in monkeys with high renin status when administered orally.

The amino acid derivative represented by general formula (I) of the present invention and its pharmacologically acceptable acid addition salt can be made into a pharmaceutical composition according to a conventional method. Examples of such pharmaceutical compositions include tablets, capsules, granules, and injections.

The amino acid derivatives and pharmacologically acceptable acid addition salts represented by the general formula (I) have a strong renin activity inhibitory effect, with a 50% inhibitory activity value in the human renin-sheep renin substrate system or in human high-renin plasma. (ICso) is 1.
2 Xl0-7 to 2.3 Xl0-8 molar concentration and 2
．． At a molar concentration of 9X10-' to 1.5X10-8, it clearly lowers the blood pressure of common marmosets in a high renin state, and has low toxicity. When a pharmaceutical composition containing the amino acid derivative represented by general formula (I) or a pharmacologically acceptable acid addition salt thereof is used for treatment, the dosage is determined based on the degree of disease, sex, age, and weight of the patient. However, oral administration is approximately 5 mg per day for adults.
~5000 mg, parenterally 1 mg per day~
It can be administered within the range of 100,100O.

〔Example〕

In order to further explain the present invention in detail, reference examples and examples are given below. Note that the melting points of the compounds in each Reference Example and Examples are uncorrected. In addition, the NMR spectra of each compound were measured using a JEOL JNM-GX270 high-resolution nuclear magnetic resonance apparatus. The mass spectrum was measured by the FAB method using a JEOL JMS-DX300 mass spectrometer. Thin layer chromatography was performed using Merck's precoated plate silica gel (precoa).
ted plates 5ilica gel)6
Column chromatography was performed using Merck's Kieselgel 60 (23
0-400 mesh). The developing solvent for thin layer chromatography is chloroform/methanol/
Using two types, the lower layer of a mixture of water = 8/3/1 and the mixture of chloroform/methanol = 5/1, the Rf value (R
f+ and Rf2) were calculated.

Reference example 1 32.3 g of ethyl succinate and 29 g of 1-naphthaldehyde
．． Dissolve 0 g in absolute ethanol 320 TlLi2,
After adding 10.7 g of 50% sodium hydride (oil-based) while cooling with water, the mixture was heated under reflux for 0.5 hour. Add 230 mA of 1N sodium hydroxide aqueous solution to this solution,
Heat to reflux for an hour. The solvent is distilled off under reduced pressure, water is added to the residue, the neutral part is extracted and removed with ether, the aqueous layer is acidified with concentrated hydrochloric acid, and extracted with ether. After washing the ether layer with saturated brine, it is dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, benzene was added to the residue, and the precipitated crystals were collected by filtration to obtain 26.5 g of 2-(1-naphthylmethylene)succinic acid as yellow crystals.

Add 260 llL12 of acetic anhydride to 24.5 g of 2-(1-naphthylmethylene)succinic acid and heat at 60°C for 1 hour. The solvent was distilled off under reduced pressure, a 1/1 mixture of benzene/hexane was added to the residue, and the precipitated crystals were collected by filtration to give 2-(1-naphthylmethylene)succinic anhydride 1 as orange-yellow crystals.
Obtain 6.0g.

1. Dissolve 348 mg of -proline methyl ester hydrochloride in 15 d of methylene chloride, and add 0.0 ml of triethylamine to this solution.
．． Add 29 mN and 500 mg of 2-(1-naphthylmethylene)succinic anhydride and stir at room temperature for 2 hours. The reaction solution was washed with dilute hydrochloric acid and saturated saline, and dried over magnesium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel flash column chromatography (elution solvent: chloroform/methanol = 15/1) to obtain a white powder of 2-(1-naphthylmethylene)-3-( 2S-methoxycarbonylpyrrolidinocarbonyl)propionic acid 66
Obtain 1 mg. Dissolve 600 mg of this propionic acid in 25 methanol and add 1.20 mg of 10% palladium on carbon.
mg and hydrogenated at normal pressure. After removing the catalyst by filtration, the solvent was distilled off under real pressure to obtain 2-(1-naphthylmethyl) as a white powder.
543 mg of -3-(2S-methquinecarbonylpyrrolidinocarbonyl)propionic acid are obtained.

Rf: 0.65 Melting point: 77-83°C IR (KBr): vco 1740.1640
Cm-'Reference Example 2 The following carboxylic acid was synthesized in the same manner as in Reference Example 1.

Pionic acid Rf: Q, 52 Melting point: 79-85°C IR (KBr): vco 1735. 1620
cm-'2-(1-naphthylmethyl)-3-(2R
-Methoxycarbo Rf: 0.65 Melting point: 62-65°C IR (KBr): vco 1735. 1640
cm-' Reference Example 3 2-(1-naphthylmethyl)-3-(23-methoxycarbonylpyrrolidinocarbonyl)propionic acid 80 mg
and boron hydroxide) Dissolve 82 mg of IJ in 2 parts of tert-butyl alcohol, 1 part of dry methanol + 1 part of dry methanol.
Add 1i2 dropwise and stir at room temperature for 30 minutes. After distilling off the solvent under reduced pressure, 1N hydrochloric acid was added to the residue to adjust the pH to 1~
2 and extracted with ethyl acetate. After washing with water, dry with anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain 72m of 2-(1-naphthylmethyl)-3-(2S-hydroxymethylpyrrolidinocarbonyl)propionic acid as a white powder.
get g.

Rfl: 0.62 Melting point: 63-67°C IR (KBr): vco 1710. 1600
cm-' Reference example 4 (2R5,3S)-3-amino-2-hydroxy-5-
N-carbobenzoxy-I5-leucinal methylhexanoate 2.81g
A solution of 3.43 g of sodium bisulfite in water was added to the mixture, and the mixture was stirred for 14 hours while cooling with water. Add a solution of 1.41 g of potassium cyanide to 50 g of water and 200 d of ethyl acetate to this reaction solution.
and stirred at room temperature for 4 hours. After washing the ethyl acetate layer with saturated brine, it is dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain 2.54 g of 3-carbobenzoxyamino-2-hydroxy-5-methylhexanenitrile as a colorless oil.

A mixture of 20 parts of dioxane and 20 parts of concentrated hydrochloric acid was added to 500 mg of cononitrile, and the mixture was heated under reflux for 12 hours. The solvent was distilled off under reduced pressure, and the remaining crystals were purified by cation column chromatography (elution solvent: 2N aqueous ammonia).
Colorless crystals of (2R3,3S)-3-amino-2-hydroxy-5-methylhexanoic acid 254 mg (2R:2
(A mixture with a S ratio of about 7:3).

Melting point: 137-140℃ IR (KBr): vco 1570 c+r
+-'NMR (020) δ: 0.8-1.0 (m, 6H), 1.2-1.
4 (m, 2H).

1.55~1.8 (+n, LH), 3.0~3t4
(m, LH).

3.89 (d, 0.7H, J=3.3Hz), 4.
00(d.

0.3H, J23.3Hz) MS: MH", 162 Reference example 5 (2R3,3S)-3-amino-2-hydroxy-5-
4.0g of methylhexanoic acid in Impropatool 50mj
2, hydrogen chloride gas is blown into the solution while stirring under water cooling, 100 d of dry benzene is added, and the mixture is heated under reflux for 10 minutes while removing the water produced. The reaction solution was concentrated to dryness under reduced pressure to obtain a white powder of (2R3,3S)-3-amino-
5.7 g of isopropyl 2-hydroxy-5-methylhexanoate hydrochloride are obtained.

IR (KBr): vco 1725 cm-'
NMR (02[1) δ: 0.8-1.1 (m, 6H), 1.29 (d
, 6H, J=6.6Hz), L5~2.0(+n,
3H), 3.6-3. '75 (m, IH), 4
．． 3-4.7 (m, LH), 5. Q ~5.2(m
, LH) Reference Example 6 The following ester was synthesized in the same manner as in Reference Example 5.

White powder IR (KBr): vco 174Q cm-
'NMR(D2[]) δ: 0.85~1.0(m, 6)1), 1.4~
1.9 (m, 3H).

3.65-3.8 (m, IH), 3.83 (s,
3H).

4,45-4.7 (m, LH) Reference example 7 (2R3,3S)-3-amino-2-hydroxy-5-
3.22g of methylhexanoic acid and 3.0g of triethylamine
8d was dissolved in 30 d of water, a dioxane 30 mA solution of 5.41 g' of 2-(tert-butyloxycarbonyloxyimino)-2-7enylacetonitrile was added, and the mixture was stirred at room temperature for 16 hours. Add 100 mI2 of water to the reaction solution.
After adding and removing the neutral part by extraction with ethyl acetate, the aqueous layer is made acidic by adding an aqueous citric acid solution and extracted with ethyl acetate. The ethyl acetate layer was washed with saturated brine and then dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain 5.10 g of (2gS, 3S)-3-tert-butyloxycarbonylamino-2-hydroxy-5-methylhexanoic acid as a pale yellow oil.

IR (KBr): vco 1710. 1675
cm-'NMR (CDCl2) δ: 0.8-1.0 (m, 6tO, 1,2-1
．． 85 (m, 12H).

3.95-4.4 (m, 2H), 4.8-5.
0 (br, IH).

9.4-10.4 (br, LH) Reference example 8 (2R3,3S)-3-tert-butyloxycarbonylamino-2-hydroxy-5-methylhexanoic acid 2
61 mg.

Dissolve 0.12 me of isoamylamine and 176 mg of 1-hydroxybenzotriazole in a mixture of N,N-dimethylformamide 2- and 2 ml of tetrahydrofuran, and add 206 mg of dicyclohexylcarbodiimide while stirring under water cooling.
mg and stirred for 16 hours. After cooling the reaction solution with ice and filtering off insoluble matter, the solvent was distilled off under reduced pressure. Dissolve the residue in ethyl acetate, add citric acid aqueous solution, 5%
After sequentially washing with an aqueous sodium hydrogen carbonate solution and a saturated saline solution, it is dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel flash column chromatography (elution solvent: chloroform).
185 mg of (2R3,3S)-3-tert-butyloxycarbonylamino-2-hydroxy-5-methylhexanoylisoamylamide is obtained in the form of a white powder. (
IR(KBr) :'Co 1700. 1635
cm-') 180 mg of the above amide was mixed with 20 mg of methanol.
Dissolve in ntf2, add 8 parts of 2N hydrochloric acid, and heat at 60°C for 1 hour. The solvent was distilled off under reduced pressure to obtain a white powder (2
138 mg of R3,3S)-3-amino-2-hydroxy-5-methylhexanoylisoamylamide hydrochloride are obtained. (IR (KBr) Niji Co 1640 cm
-':]Reference Example 9 The following amine was synthesized in the same manner as in Reference Example 8.

Reference example 10 Isopropyl cuxi-5-methylhexanoate 2p-L
- Suspend 10.0 g of histidine methyl ester dihydrochloride in 200 mg of dry chloroform, add 18.4 mA of triethylamine and 10.2 g of 4-methoxybenzyloxycarbonyl azide under cooling, and stir at 0°C for 16 hours. The solvent was distilled off under reduced pressure, a 5% aqueous sodium bicarbonate solution was added to the residue, extracted with ethyl acetate, washed with water, and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the residue was subjected to silica gel flash column chromatography (elution solvent: chloroform/methanol = 1
0/1) to obtain 11.0 g of yellow oily N-(4-methoxybenzyloxycarbonyl)-L-histidine methyl ester. 10.9 g of this ester was dissolved in 112 mN of methanol, and 9.9 g of hydrazine 1 permanent was dissolved in 112 mN of methanol.
Add mj2 and stir at room temperature for 4 hours. The solvent was distilled off under reduced pressure, the residue was washed with ethanol, and then evaporated under reduced pressure for 40 min.
Dry at below ℃ to obtain N-(4-methoxybenzyloxycarbonyl)-L-histidine hydrazide 4 in the form of white powder.
．． Obtain 9g.

485 mg of this hydrazide was suspended in 5 rnl of N,N-dimethylformamide, and mixed with 5.1N dry hydrogen chloride/0.9 N,N-dimethylformamide with stirring at -20°C.
0d followed by 0.23n+j2 of isoamyl nitrite. After confirming the disappearance of hydrazide, the temperature of the reaction solution was lowered to -30°C, neutralized with 0.67 mA of triethylamine, and N-(4-methoxybenzyloxycarbonyl)-
Prepare a cold solution of histidine azide. Separately (2R3
,3S)-3-Amino-2-hydroxy-5-methylhexanoic acid isopropyl hydrochloride (350 mg) and triethylamine (0.46 mA) in dry N,N-dimethylformamide (8 mA) was cooled with water, and the above cold azide solution was added dropwise. ,1
Stir for 6 hours. The solvent was distilled off under reduced pressure, and the residue was
% hydrogen carbonate) is added and extracted with ethyl acetate, then washed with saturated brine and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel flash column chromatography (elution solvent: chloroform/methanol = 20/1).
325 mg of isopropyl N-(4-methoxybenzyloxycarbonyl)-sehistidyl-(2R3,3S)-3-amino-2-hydroxy-5-methylhexanoate in the form of a white powder is obtained. 320 mg of this isopropyl hexanoate is dissolved in 46 mN of isopropyl alcohol, 300 mg of p-)reninsulfonic acid (anhydride) and 48 mg of 10% palladium on carbon are added, and the mixture is hydrogenated at normal pressure. After removing the catalyst by filtration, the solvent was distilled off under reduced pressure to obtain (2R3,3S)-3-(L-histidyl)amino-2 as a white powder.
-Isopropyl-hydroxy-5-methylhexanoate 2
513 mg of p-toluenesulfonate.

Rf: 0.09 IR(KBr): νco 1720. 1680
cm-'Reference Example 11 The following amino acid derivative was synthesized in the same manner as in Reference Example 10.

White powder Rfl: 0.15 IR (KBr): vco 158Q cm-'
Dihydrochloride white powder Rf: 0.16 IR (KBr): vco 155Q c+y
r'White powder RL: 0.29 IR(KBr): L/CO1680Cm-'Example 1 2-(1-naphthylmethyl)-3-(2S-methoxycarbonylpyrrolidinocarbonyl)propionic acid 346
mg and 3-L-histidylamino-2-hydroxy-5
- Dissolve 2p-toluenesulfone in isopropyl methylhexanoate, add 0 diphenyl phosphate azide under stirring under water cooling,
Add 24mj2 and triethylamine 0.43-
Leave as is - stir overnight. The solvent was distilled off under reduced pressure, a 5% aqueous sodium hydrogen carbonate solution was added to the residue, and the mixture was extracted with ethyl acetate. After washing with water, it is dried over anhydrous magnesium sulfate, and the solvent is distilled off under reduced pressure. The residue was purified by silica gel column chromatography (elution solvent: chloroform/methanol = 30/1) to obtain a white powder (2RS.
)-3- (N- C2- (1-naphthylmethyl)-
Isopropyl 3-(2S-methoxycarbonylpyrrolidinocarbonyl)propionyl]-histidyl)amino-2-hydroxy-5-methylhexanoate 117m
get g.

Melting point: 90-94°C Rfl: 0.68 Rf2: 0.65 MS: MH”, 692 Example 2 Propyl 2-(1-naphthylmethyl)-3-(4-hydroxy-
17 mg of 2S-methoxycarbonylpyrrolidinocarbonyl)propionic acid and isopropyl 3-histidylamino-2-hydroxy-5-methylhexanoate 2p-
) 28 mg of renin sulfonate was dissolved in 2 ml of N,N-dimethylformamide, and 0.012 ml of diphenylphosphoryl azide and 0.01 ml of triethylamine were added under stirring while cooling with water. Add 021 fart and stir overnight. The solvent was distilled off under reduced pressure, a 5% aqueous sodium hydrogen carbonate solution was added to the residue, and the mixture was extracted with ethyl acetate. After washing with water, it is dried over anhydrous magnesium sulfate, and the solvent is distilled off under reduced pressure. The residue was subjected to preparative silica gel thin layer chromatography (developing solvent: chloroform/methanol = 5/1) to isolate and purify the portion corresponding to Rf2 of 0.63, yielding white powder (2RS, 3S)-3. - (N-C2- (
1-naphthylmethyl)-3-(4-hydroxy-2S-
methoxycarbonylpyrrolidinocarbonyl)propionyl] -L- histidyl)amino-2-hydroxy-
28 mg of isopropyl 5-methylhexanoate is obtained.

Melting point: 105-108°C Rf+: o. 65 Rf2: 0.63 MS: MH”, 708 Example 3 31 mg of 2-(1-naphthylmethyl)-3-(2-methoxycarbonylpyrrolidinocarbonyl)propionic acid and 3-(L-histidyl)amino-2- Hydroxy-5-
40 mg of methylhexanoate dihydrochloride was suspended in 3 parts of N,N-dimethylformamide, and 0.022 mff1 of diphenylphosphoric acid azide and 0.038 mff of triethylamine were added to the suspension while stirring under water cooling, and the mixture was stirred for 16 hours.

The solvent was distilled off under reduced pressure, and a 5% aqueous sodium bicarbonate solution was added to the residue, extracted with ethyl acetate, washed with saturated brine, and dried over anhydrous magnesium sulfate.

The solvent was distilled off under reduced pressure, and the residue was purified by preparative silica gel thin layer chromatography (developing solvent: chloroform/methanol = 5/1) to isolate and purify the portion corresponding to Rf2 of 0.48, which gave a white powder. (2R3,3S)-3
-(N-C2-(1-naphthylmethyl)-3-(2S-
methoxycarbonylpyrrolidinocarbonyl)propionyl]-cy-histidyl)amino-2-hydroxy-5
-13 mg of methylhexanoate are obtained.

Melting point: 102-107°C Rt: 0.60 tu2: 0.48 M5: MH", 664 Example 4 2-(1-naphthylmethyl)-3-(2-methoxycarbonylpyrrolidinocarbonyl)propionic acid 37 mg and 3-(L-histidyl)amino-2-hydroxy-5-
Methylhexanoylisoamylamide dihydrochloride 44mg
was suspended in 3nd N,N-dimethylformamide, 0.03 ml of diphenylphosphoric azide and 0.05 d of triethylamine were added while stirring under water cooling, and the mixture was stirred for 16 hours. The solvent was distilled off under reduced pressure, and a 5% aqueous sodium bicarbonate solution was added to the residue, extracted with ethyl acetate, washed with water, and dried over anhydrous magnesium sulfate.

The solvent was distilled off under reduced pressure, and the residue was subjected to preparative silica gel thin layer chromatography (developing solvent: chloroform/methanol = 5/1) to isolate and purify the portion corresponding to Rf2 of 0.51, resulting in a white powder. (2R3,3S)-
3-(N-C2-(1-naphthylmethyl)-3-(2S
-methoxycarbonylpyrrolidinocarbonyl)propionyl]=shi-histidyl)amino-2-hydroxy-5
-13 mg of methylhexanoylisoamylamide are obtained.

Melting point = 103-108°C Rf: 0.65 Rf2: 0.51 M5: MH”, 719 Example 5 2-(1-naphthylmethyl)-3-(2S-methoxycarbonylpyrrolidinocarbonyl)propionic acid 20 mg
and L-histidyl-statyl isoamylamide 2p-
40 mg of toluene sulfonate was dissolved in 2 d of N,N-dimethylformamide, and 0.014 IrLi2 of diphenylphosphoric acid azide and 0.025 of triethylamine were added to the mixture while stirring under water cooling, and the mixture was stirred overnight. The solvent was distilled off under reduced pressure, a 5% aqueous sodium hydrogen carbonate solution was added to the residue, and the mixture was extracted with ethyl acetate. After washing with water, it is dried over anhydrous magnesium sulfate, and the solvent is distilled off under reduced pressure. The residue was subjected to preparative silica gel thin layer chromatography (developing solvent: chloroform/methanol = 5/1) to obtain Rf
The part corresponding to 0.66 was isolated and purified to obtain white powdery N-C2-(1-naphthylmethyl)-3-(2S-methoxycarbonylpyrrolidino)carbonyl]-L-histidyl rustatyl. 15 mg of isoamylamide are obtained.

Melting point = 80-85°C Rf: 0.68 Rf2: 0.66 M5: MH”, 733 Example 6 2-(1-naphthylmethyl)-3-(2S-hydroxymethylpyrrolidinocarbonyl)propionic acid 21 mg 40 mg of isopropyl 1-histidylamino-2-hydroxy-5-methylhexanoate was dissolved in 2 mN of N,N-dimethylformamide, and 0.016 g of diphenyl phosphoryl azide was dissolved under stirring under water cooling.
Add mN and 0.028 ml of triethylamine and stir overnight. The solvent was distilled off under reduced pressure, a 5% aqueous sodium hydrogen carbonate solution was added to the residue, and the mixture was extracted with ethyl acetate. After washing with water, dry with anhydrous magnesium sulfate,
The solvent is distilled off under reduced pressure. The residue was subjected to preparative silica gel thin layer chromatography (developing solvent: chloroform/methanol = 5/1) to isolate and purify the portion corresponding to Rf2 of 0.59, and white powder (2R3,3S) was obtained.
-3-(N-C2-(1-naphthylmethyl)-3-(2
S-hydroxymethylpyrrolidinocarbonyl)propionyl]-histidyl)amino-2-hydroxy-5
-16 mg of isopropyl methylhexanoate are obtained.

Melting point 2 86-92°C Rf, + 0.60 Rf2: 0.59 M5: MH”, 664 Example 7 2-(1-naphthylmethyl)-3-(2R-methoxycarbonylpyrrolidinocarbonyl)propionic acid 20 mg
and 39 mg of isopropyl 3-L-histidylamino-2-hydroxy-5-methylhexanoate 2p-toluenesulfonate were mixed with 5 mA of N,N-dimethylformamide.
Diphenyl phosphoryl azide 0.0
Add 0.025 mA of 14- and triethylamine and stir overnight. The solvent was distilled off under reduced pressure, a 5% aqueous sodium hydrogen carbonate solution was added to the residue, and the mixture was extracted with ethyl acetate. After washing with water, dry with anhydrous magnesium sulfate,
The solvent is distilled off under reduced pressure. The residue was subjected to bleparative silica gel thin layer chromatography (developing solvent: chloroform/methanol = 5/1) to isolate and purify the portion corresponding to Rf2 of 0.56, and white powder (2R3,3S) was purified.
-3-(N-C2-(1-naphthylmethyl)-3-(2
R-methoxycarbonylpyrrolidinocarbonyl)propionyl]-histidyl)aminol 2-hydroxy-
Melting point to obtain 7 mg of isopropyl 5-methylhexanoate: 99-103°C Rfl: 0.62 Rf2 + 0.56 M5 + MH”, 692 Example 8 Inhibition of renin activity with human renin-sheep renin substrate pH 7,4 125 mM pyrophosphate buffer 20
0 reverse and 20mM as angiotensin converting enzyme inhibitor
Noshiichi 25d aqueous solution of phenylalanyl-L-alanyl-L-proline, 50% partially purified sheep renin substrate containing 2000ng angiotensin I equivalent/d, deionized water 150%
Reverse and dimethyl sulfoxide solution of the compound of the present invention 50d
or dimethyl sulfoxide 50 as a control group.
20-30 ng angiotensin I/in solution of d.
Add 25 replicates of purified human renin at ml/hr and incubate at 37°C.
(incubate for 15 minutes in a water bath)
e) After that, place the reaction solution in a 100°C water bath for 5 minutes to stop the reaction.

After cooling, 200 d was collected and the amount of angiotensin produced by the addition of renin was measured by radioimmunoassay.
(radioimmuno assay) method,
Inhibitory activity was determined using the following formula.

Inhibitory Activity (%) - Control The 50% inhibitory activity molar concentration (ICso') was determined from the inhibitory activity determined by the above formula of the compound of the present invention, and the results are shown below.

Compound name IC5o value (mol) Compound name
IC50 value (mol) Example 9 Inhibitory effect on renin activity in human high-renin plasma 14 mM BDTA 2 Na in 500 volumes of human high-renin plasma
and 0.3% neomycin sulfate at pH 7.0.
5M phosphate buffer
er) 350m, 50ml of an aqueous solution of 20mM phenylalanyl-mono-alanyl-proline as an angiotensin-converting enzyme inhibitor, and 100 doses of a dimethyl sulfoxide solution of the compound of the present invention or 100 d of dimethyl sulfoxide as a control group. . Of this reaction solution, 200 copies were placed in a 4°C water bath.
Incubate the remaining 800 ml in a 37°C water bath for 60 minutes.

2004 is taken from the reaction solution incubated at 37°C, immediately cooled on ice, and the amount of angiotensin is quantified by radioimmunoassay along with the reaction solution incubated in an ice bath.

Plasma renin activity is calculated by subtracting the amount of angiotensin II in the reaction solution incubated at 4°C from the amount of angiotensin II in the reaction solution incubated at 37°C. Inhibitory activity (%) was determined by the following formula.

Inhibitory Activity (%) - Control The 50% inhibitory activity molar concentration (ICso) was determined from the inhibitory activity determined by the above formula of the compound of the present invention, and the results are shown below.

Compound name IC5o value (mol) Sufunal 4
SoT':, Lua (r: Compound name IC5o value (mol) Example 1
0 Blood pressure lowering effect on common marmosets Clinically conducted by Hofbauer et al.
AND EXPERIMENTAL HYPERTHEASE:
/ [(CIin, Snake xp, Hypertens,)
Volume A5, No. 7-8, Pages 1237-1247, 198
The common marmoset (comm
Furosemide 15. mg/kg/day every other day 3
Administer twice to create an artificially high renin state. Blood pressure was measured consciously at the third molar after discontinuation of furosemide administration.

Measurement method: Male common marmosets weighing 390 g and 365 g are consciously fixed on a small monkey fixing table, and the arterial pressure is measured using a non-invasive blood pressure measuring device. The compound of the present invention is dissolved in dilute hydrochloric acid and administered orally using a gastric tube. The results of the 100 mg/kg administration example are shown below.

100 mg/kg oral administration (2 cases) mean blood pressure (
mmHg) Control 94.0 1 hour after administration 77.5 2 hours 74.0 3 hours 70.0 5 hours 92.2 7 hours 101.5 [Effects of the invention] The compound represented by the general formula (I) of the present invention Amino acid derivatives and their pharmacologically acceptable acid addition salts are human renin-
50% in renin activity inhibition test using sheep renin substrate system
Inhibitory activity value (ICso) is 1.2 x 10-7 to 2.3
X 10-8 molar concentration, and also showed a 50% inhibitory activity value (
ICso) is 2.9 x 10-7 to 1.5 x 1
It exhibits a strong renin activity inhibitory effect at a concentration of 0-8 molar,
and has low toxicity. Furthermore, the amino acid derivatives represented by the general formula (I) of the present invention and their pharmacologically acceptable acid addition salts are stable against proteolytic enzymes such as chymotrypsin and pepsin, and are stable against oral administration. can lower blood pressure in monkeys with high renin status.

Therefore, the amino acid derivatives represented by general formula (I) of the present invention and their pharmacologically acceptable acid addition salts are useful as orally administrable antihypertensive agents.

Claims (1)

[Claims] 1) General formula ▲ Numerical formula, chemical formula, table, etc. ▼ (In the formula, X is a lower alkoxycarbonyl group or a hydroxymethyl group, Y is a hydrogen atom or a hydroxyl group, and His is L- is a histidyl group, n is 0 or 1, Z is -O- or -NH-, and R has 1 carbon number.
~7 linear or branched alkyl groups) and pharmacologically acceptable acid addition salts thereof. 2) The claim set forth in claim 1 expressed by the general formula ▲ includes mathematical formulas, chemical formulas, tables, etc. ▼ (X, Y, His, n, Z and R in the formula have the same meanings as above) Amino acid derivatives and their pharmacologically acceptable acid addition salts. 3) General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (C^* in the formula indicates S configuration, X, Y, His, n,
3. Amino acid derivatives and pharmacologically acceptable acid addition salts thereof according to claim 2, wherein Z and R have the same meanings as above. 4) Claim 3 expressed by the general formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (X, Y, His, C^*, Z and R in the formula have the same meanings as above) The described amino acid derivatives and their pharmacologically acceptable acid addition salts. 5) Claim 3 represented by the general formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (X, Y, His, C^*, Z and R in the formula have the same meanings as above) The described amino acid derivatives and their pharmacologically acceptable acid addition salts. 6) Formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (His and C^* in the formula have the same meaning as above)
The amino acid derivative according to claim 4 represented by: and a pharmacologically acceptable acid addition salt thereof. 7) Formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (His and C^* in the formula have the same meaning as above)
The amino acid derivative according to claim 4 represented by: and a pharmacologically acceptable acid addition salt thereof. 8) Formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (His and C^* in the formula have the same meaning as above)
The amino acid derivative according to claim 4 represented by: and a pharmacologically acceptable acid addition salt thereof. 9) Formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (His and C^* in the formula have the same meaning as above)
The amino acid derivative according to claim 4 represented by: and a pharmacologically acceptable acid addition salt thereof. 10) Formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (His and C^* in the formula have the same meanings as above)
The amino acid derivative according to claim 4 represented by: and a pharmacologically acceptable acid addition salt thereof. 11) Formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (His and C^* in the formula have the same meaning as above)
The amino acid derivative according to claim 5, represented by: and a pharmacologically acceptable acid addition salt thereof.