JPH0532601A - Intermediate of substituted peptide compound - Google Patents

Abstract

PURPOSE: To provide a new substituted peptide compd. and a new intermediate for preparing of its salts useful for suppression agent of blood pressure.

CONSTITUTION: The compd. represented by formula I [where R is H, Prot, a lower alkyl, cycloalkyl, the group represented by formula II ((m) is 0-4), -(CH₂)₂N(H)-Prot, etc.; R₁ is H, a lower alkyl, halo-substd. alkyl, the group represented by formula III, -(CH₂)_r-S-Prot ((r) is 1-4), -(CH₂)_r-O-Prot, etc., provided that R₁ is H when R is exept H (Prot is easily eliminable group); R₂ is the group represented by formula IV ((m) is 0-4), etc.; R₃ is H, an alkyl, halo-sustd. alkyl, the group represented by formula IV, etc.], e.g. (±)-1-[N-[3-(benzoylamino)-2-oxo-4-phenylbutyl]-N-methyl-glycyl]-L-proline- monohydrochloride. The compd. represented by formula I is obtained by coupling an aminoketone-carboxylic acid represented by formula V with a compd. represented by HX [X is the group represented by formula VI (R₆ is H, an alkyl, etc.; R₇ is H, an alkyl, OH, etc.)] in the presence of coupling agent.

COPYRIGHT: (C)1993,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an intermediate of a substituted peptide compound, and more particularly to an intermediate for producing a novel substituted peptide compound having pharmacological activity and salts thereof.

[0002]

The intermediates according to the present invention have the formula:

[Chemical 12] Embedded image By using such a compound [II], a substituted peptide compound represented by the following formula [I] and salts thereof can be produced:

[Chemical 13] [Where X is

[Chemical 14]

[Chemical 15] Alkyl), azido, amino,

[Chemical 16] 1- or 2-naphthyl represented by,-(CH ₂ ) m-
(Cycloalkyl),

[Chemical 17] 1- or 2-naphthyloxy represented by
(Lower alkyl),

[Chemical 18] 1- or 2-naphthylthio represented by: R ₈ is keto,
halogen,

[Chemical 19] 1- or 2-naphthyloxy represented by
(Lower alkyl),

[Chemical 20] 1- or 2-naphthylthio represented by: R ₉ is keto or

[Chemical 21] R ₁₀ is halogen or —Y—R ₁₆ ; R ₁₁ , R ′ ₁₁ , R ₁₂ and R ′ ₁₂ are each independently hydrogen or lower alkyl,
Alternatively, R ′ ₁₁ , R ₁₂ and R ′ ₁₂ are hydrogen and R ₁₁ respectively.
Is

[Chemical formula 22] R ₁₃ is hydrogen, lower alkyl having 1 to 4 carbons, 1 to 1 carbons
4 lower alkoxy, alkylthio having 1 to 4 carbon atoms, chloro, bromo, fluoro, trifluoromethyl, hydroxy, phenyl, phenoxy, phenylthio or phenylmethyl; R ₁₄ is hydrogen, lower alkyl of 1 to 4 carbon atoms, carbon atoms 1 to 4 lower alkoxy, 1 to 4 carbon lower alkylthio, chloro, bromo, fluoro, trifluoromethyl or hydroxy; m is 0, 1, 2, 3 or 4; p is 1, 2 or 3 (however, When either R ₁₃ or R ₁₄ is hydrogen, methyl, methoxy, chloro or fluoro, p is an integer greater than 1); R ₁₅ is hydrogen or lower alkyl having 1 to 4 carbons; Y is oxygen or sulfur. R ₁₆ is lower alkyl having 1 to 4 carbon atoms or

[Chemical formula 23] Alternatively, an unsubstituted 5- or 6-membered cyclic group formed by combining two R ₁₆ or a lower alkyl substituent having 1 to 4 ring-forming carbon atoms having 1 to 4 carbon atoms or di (lower (carbon Substituted 5- or 6-membered cyclic group having an alkyl) substituent of the formulas 1 to ₄ ; R ₄ is hydrogen, lower alkyl,

[Chemical formula 24] R ₅ is hydrogen, lower alkyl,

[Chemical 25] r is an integer from 1 to 4; R ₁₉ is lower alkyl, benzyl or phenethyl; R ₂₀ is hydrogen, lower alkyl, benzyl or phenethyl; R is hydrogen, lower alkyl, cycloalkyl,

[Chemical formula 26] _{, - (CH 2) 2 -NH} 2, - (CH 2) 3 -NH 2, - (CH 2) 4
_{-NH 2, - (CH 2)} 2 -OH, - (CH 2) 3 -OH, - (C
_{H 2) 4 -OH, - (} CH 2) 2 -SH, - (CH 2) 3 -SH , or _{- (CH 2) 4 -SH;} R 1 is hydrogen, lower alkyl, halo-substituted (lower) alkyl,

[Chemical 27] (However, when R is a group other than hydrogen, R ₁ is hydrogen); R ₂
Is

[Chemical 28] R ₃ is hydrogen, lower alkyl,

[Chemical 29]

[Chemical 30] (m, R ₁₄ , p and r are as defined above); R ₁₇ is hydrogen, lower alkyl, cycloalkyl or phenyl, R ₁₈ is hydrogen, lower alkyl, lower alkoxy or phenyl, or R ₁₇ and R ₁₈ are combined to form — (CH ₂ ) ₂ ,
_{- (CH 2) 3 -,} - CH = CH- or

[Chemical 31] R ₂₁ and R ₂₂ each independently represent hydrogen or lower alkyl; R ₂₃ represents lower alkyl].

In the formula [I], the terms used to define various symbols have the following meanings. Lower alkyl has 7 carbon atoms
Lower alkyl, which includes straight chain or branched groups not exceeding 4 and does not exceed 4 carbon atoms, is most preferable, and methyl and ethyl are most preferable. Similarly, lower alkoxy and lower alkylthio include lower alkyl as described above attached to oxygen or sulfur.

Cycloalkyl includes saturated cyclic groups of 3 to 7 carbon atoms, of which cyclopentyl and cyclohexyl are most preferred.

Halogen includes chloro, bromo and fluoro.

Halo-substituted (lower) alkyl means a lower alkyl group as defined above in which one or more hydrogen has been replaced by chloro, bromo or fluoro, such as trifluoromethyl (which is preferred), pentafluoroethyl,
Includes 2,2,2-trichloroethyl, chloromethyl, bromomethyl and the like.

[0007]

[Chemical 32] Represents that each alkylene bridge can be bonded to all ring-forming carbon atoms.

Next, a method for producing the compound [I] will be described. The compound [I] has the formula:

[Chemical 33] [Wherein R, R ₁ , R ₂ and R ₃ have the same meanings as defined above] and an aminoketonecarboxylic acid (intermediate of the present invention) [II],
formula:

[Chemical 34] [In the formula, R _{6 in} X represents a protecting group which can be easily removed], or an amino acid (or imino acid) ester [II
[I] can also be obtained by a coupling treatment in the presence of a coupling agent such as dicyclohexylcarbodiimide. The compound [I] in which R ₆ is hydrogen can be obtained by removing the R ₆ protecting group.

The aminoketonecarboxylic acid intermediate [II] is
formula:

[Chemical 35] [Wherein R, R ₂ and R ₃ have the same meanings as defined above] and an aminoketone [IV]:

[Chemical 36] [In the formula, prot represents an easily protecting ester protecting group such as t-butyl. halo and R ₁ are as defined above]
By reacting a haloacetic acid ester [V] represented by

[Chemical 37] [Wherein R ₁ , R ₂ , R ₃ and prot have the same meanings as described above], and the ester protecting group can be removed by preparing an ester [VI].

The above aminoketone [IV] can be produced as follows. formula:

[Chemical 38] [Wherein R ₄₀ represents a protecting group such as benzyloxycarbonyl. R has the same meaning as above] for converting a carboxyalkylamine [VII] into, for example, its acid chloride, and then with this formula:

[Chemical Formula 39] [Wherein R ₂ and R ₃ have the same meanings as above] are reacted with an oxazolone [VIII] to give the following formula:

[Chemical 40] [Wherein R, R ₂ , R ₃ and R ₄₀ have the same meanings as described above] are obtained. The R ₄₀ protecting group of this compound [IX] is
The aminoketone [IV] can be obtained by elimination by a reduction method such as hydrogenation.

An aminoketone in which R is a group other than hydrogen
[IV] has the formula:

[Chemical 41] [In the formula, halo represents halogen (preferably chloro or bromo). R ₂ and R ₃ have the same meaning as above] and a ketone [X] and a formula:

[Chemical 42] [Wherein R is as defined above] can be obtained by reacting a substituted amine.

In the various reactions described above, R, R
Any one or all of ₁ , R ₃ and R ₅

[Chemical 43] If, the hydroxy, amino, imidazolyl, mercapto or guanidinyl functional groups in these groups should be protected during the reaction. Suitable protecting groups include benzyloxycarbonyl, t-butoxycarbonyl, benzyl,
Includes benzhydryl, trityl, etc., and nitro (for guanidinyl). The protecting group is removed by, for example, hydrogenation, treatment with acid or other known method to achieve reaction completion.

Compound [I] and its pharmacologically acceptable salts are hypotensive agents. These compounds are decapeptides angiotensin I angiotensin II
It is useful in inhibiting the conversion to and thus reducing or ameliorating angiotensin-related hypertension. Angiotensin I is produced by the action of the enzyme renin on angiotensinogen and blood pseudoglobulin. Angiotensin I is angiotensin converting enzyme (ACE)
To angiotensin II. Angiotensin II is a blood pressure-increasing active substance,
Relevant as a causative agent of various types of hypertension (eg humans). Compound [I] intervenes in the angiotensinogen → (renine) → angiotensin I → angiotensin II system by suppressing angiotensin converting enzyme and reducing or blocking the production of angiotensin II, a blood pressure-increasing active substance. Thus, the compound [I]
By administering a composition containing one (or a mixture), angiotensin-dependent hypertension in a hypertensive mammal (eg, human) can be reduced. To lower blood pressure, about 0.1-100 mg (preferably about 1-5)
It is appropriate to administer the dose of 0 mg) / kg (body weight) / day once a day, preferably in 2 to 4 divided doses. The active substance is preferably administered orally, but it may also be administered parenterally, such as subcutaneously, intramuscularly, intravenously or intraperitoneally.

[0014] (Enkephalinase) inhibitory activity and useful as an analgesic. Therefore, the pain of mammals can be alleviated by administering a composition containing one or more mixtures of the compound [I] or a pharmaceutically acceptable salt thereof. Active compound about 0.
1 to 100 mg (preferably about 1 to 50 mg) / kg (body weight) / day is used as a standard, and the desired analgesic activity is exhibited by administering this once daily, preferably in 2 to 4 divided doses. Forget The composition is preferably administered orally, but may be administered parenterally, such as subcutaneously.

[0015]

EXAMPLES Next, the production method of the preferred compound of the present invention will be specifically described with reference to examples. LH-20 is pharmaci
a Sephadex chromatography gel, commercially available from Fine Chemicals. The temperature (° C.) in the examples may be simply expressed as “°”.

Reference Example 1 Preparation of 1- (bromoacetyl) -L-proline.1,1-dimethylethyl ester: -L-proline dimethylethyl ester (34.2 g, 0.2) in methylene chloride (250 ml). Mole) cooling (-10
°) solution, diisopropylethylamine (38.3 ml,
0.22 mol) and bromoacetyl chloride (16.5 ml,
0.2 mol) is added dropwise over 20 minutes while maintaining the temperature between -10 ° and -5 °. The dark reaction mixture is stirred overnight (-10 ° to room temperature) and concentrated under reduced pressure. The oily residue was redissolved in ethyl acetate and washed with saturated sodium bicarbonate (2
Washed with 10% potassium bicarbonate (2 times) and water (2 times), dried (Na ₂ SO ₄ ) and concentrated to a dark oily residue (28.
0g). Flash chromatography (LPS-
1 Silica gel, 10% ethyl acetate / methylene chloride) gives 11.0 g of 1- (bromoacetyl) -L-proline.1,1-dimethylethyl ester as a pale yellow oil.

Example 1 Preparation of (±) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -L-proline monohydrochloride: -a ) Preparation of [3- (benzoylamino) -2-oxo-4-phenylbutyl] methylcarbamic acid phenylmethyl ester: -N-methyl-N-[(phenylmethoxy) carbonyl] glycine (2.23 g, 10 mmol) Dissolve in 30 ml of tetrahydrofuran and cool in an ice bath. Oxalyl chloride (1m
1, 11.5 mmol) followed by 2 drops of dimethylformamide. After stirring for 30 minutes in an ice bath, the mixture is stirred at room temperature for 1 hour. To this is added 0.25 ml oxalyl chloride. The mixture is evaporated, redissolved in 15 ml tetrahydrofuran and stirred in an ice bath. 2-phenyl-4-
(Phenylmethyl) -5 (4H) -oxazolone (3.1 g,
A solution of 12.4 mmol) in 15 ml of tetrahydrofuran is added to the above solution and stirred in an ice bath. Triethylamine (1.4 ml, 10 mmol) is added and the solution is stirred at room temperature overnight. The precipitated triethylamine hydrochloride is filtered off. Tetrahydrofuran was removed from the residue, and then redissolved in pyridine (5 ml), p-dimethylaminopyridine
(20 mg) is added. After stirring for 3 hours at room temperature, acetic acid (5 ml) is added and the reaction mixture is kept at 105 ° for 30 minutes. Then the reaction mixture is evaporated down, the residue is dissolved in ethyl acetate and washed with aqueous sodium bicarbonate solution and water. After trituration with ethyl acetate / hexane, 2.2 g of homogeneous [3- (benzoylamino) -2-oxo-4-phenylbutyl] methylcarbamic acid phenylmethyl ester are obtained. mp
140-141 °.

B) (±) -N- [3- (methylamino) -2-
Oxo-1- (phenylmethyl) propyl] benzamide
Preparation of Hydrochloride: Ethanol (50 ml) containing 1N-hydrochloric acid (2 ml), [3-
(Benzoylamino) -2-oxo-4-phenylbutyl] methylcarbamic acid phenylmethyl ester (0.5
g) Dissolve. Palladium / carbon catalyst (10%, 100 mg)
And hydrogenation is continued overnight. The reaction mixture is then filtered, evaporated, dissolved in water, lyophilized to 300 mg.
(±) -N- [3- (methylamino) -2-oxo-1-
(Phenylmethyl) propyl] benzamide hydrochloride is obtained as a homogeneous white powder.

C) Preparation of (±) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -L-proline.1,1-dimethylethyl ester : (±) -N- [3- in dimethylformamide (20 ml)
(Methylamino) -2-oxo-1- (phenylmethyl) propyl] benzamide · hydrochloride (1.65 g, 5 mmol), 1- (bromoacetyl) -L-proline · 1,1 of Reference Example 1
-Dimethylethyl ester (2.9 g, 10 mmol) and diisopropylethylamine (1.74 ml, 10 mmol)
The reaction mixture of is stirred at room temperature under nitrogen overnight. The reaction mixture is partitioned between water / ethyl acetate and the aqueous layer is extracted again with ethyl acetate. Dry the ethyl acetate extract (Na ₂ SO ₄ ).
And concentrated to a yellow oily residue (4.0 g). Flash chromatography (150 g of Merck silica gel 6
0), 0.7 g of (±) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -L-proline.1,1-dimethyl The ethyl ester is obtained as a yellow foam.

D) Preparation of (±) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -L-proline monohydrochloride: -above The ester product of c) (0.4 g, 0.79 mmol) is treated with 2N hydrochloric acid / acetic acid (5 ml). After stirring for 45 minutes at room temperature, the reaction mixture was concentrated under reduced pressure and the resulting oily residue was triturated with ether (3 times) to give 0.28 g.
(±) -1- [N- [3- (Benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -L-proline monohydrochloride is obtained as a white solid. mp125-1
30 °. Rf = 0.73 (silica gel, n-butanol /
Acetic acid / water = 3: 1: 1). Elemental analysis (as C ₂₅ H ₂₉ N ₃ O ₅ · HCl · 0.23H ₂ O) Calculated value: C, 61.01, H, 6.24, N, 8.54 Measured value: C, 61.01, H, 6.10, N, 8.36

N- [N- [3- (benzoylamino) -2-
Oxo-4-phenylbutyl] -N-methylglycyl]-
Preparation of N-cyclohexylglycine monohydrochloride: -a) (±) -N- [3- (benzoylamino) -2-oxo-
Preparation of 4-phenylbutyl] -N-methylglycine.1,1-dimethylethyl ester :-( ±) -N- [3-prepared as described in Example 1 (b) in dimethylformamide (20 ml). (Methylamino) -2-
Oxo-1- (phenylmethyl) propyl] benzamide
To a solution of (5.0 g, 15 mmol), bromoacetic acid.1,1-
Dimethyl ethyl ester (13.8 g, 3.15 ml, 1
9.5 mmol) and diisopropylethylamine (2.
5 g, 3.4 ml, 19.5 mmol) are added. After stirring overnight at room temperature, the reaction mixture was poured into water (100 ml) and washed with ethyl acetate.
Extract with (3 times). The combined ethyl acetate extracts were washed with saturated sodium bicarbonate (2 times), 10% potassium bicarbonate (2 times), and water (2 times), dried (Na ₂ SO ₄ ) and concentrated to a yellow oil. , Which was dried under high vacuum to give a dry foam which gave 5.4 g of (±) -N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycine. 1,1-Dimethylethyl ester is obtained.

B) (±) -N- [3- (benzoylamino)-
Preparation of 2-oxo-4-phenylbutyl] -N-methylglycine monohydrochloride:-2% of the ester product of (a) above (4.51 g, 11 mmol)
Treat with N-hydrochloric acid / acetic acid (20 ml). After stirring at room temperature for 2.5 hours, the reaction mixture was concentrated under reduced pressure and the oily residue was triturated with ether to give 3.3 g of (±) -N- [3- (benzoylamino) -2-oxo-4-phenyl. Butyl]-
N-methylglycine monohydrochloride is obtained as an off-white solid. c) N- [N- [3- (benzoylamino) -2-oxo-
Preparation of 4-phenylbutyl] -N-methylglycyl] -N-cyclohexylglycine.1,1-dimethylethyl ester :-( ±) -N- [3- in distilled tetrahydrofuran (50 ml).
(Benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycine.hydrochloride (1.0 g, 2.6 mmol) was added to a solution of N-cyclohexylglycine.1,1-dimethylethyl ester (0 0.55 g, 2.6 mmol), hydroxybenzotriazole hydrate (0.39 g, 2.6 m)
Mol) and dicyclohexylcarbodiimide (0.55
g, 2.6 mmol) is added. The reaction mixture was stirred overnight,
The precipitated dicyclohexylurea is filtered off and the filtrate is concentrated. The residue was dissolved in ethyl acetate (50 ml), saturated potassium bicarbonate (2 times), 10% potassium bicarbonate (2 times), and water.
Washed (twice), dried (Na ₂ SO ₄ ) and oily residue (1.5 g).
Concentrate to. By flash chromatography (100 g Merck silica gel 60) 0.49 g N- [N-
[3- (Benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -N-cyclohexylglycine.1,1-dimethylethyl ester is obtained as a foam.

D) N- [N- [3- (benzoylamino)-
Preparation of 2-oxo-4-phenylbutyl] -N-methylglycyl] -N-cyclohexylglycine monohydrochloride:
The ester product of (c) above (0.48 g, 0.87 mmol)
Is treated with 2N hydrochloric acid / acetic acid (10 ml). After stirring at room temperature for 2 hours, the reaction mixture was concentrated under reduced pressure and the oily residue was triturated with ether overnight to give 0.32 g of N- [N-
(3- (Benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -N-cyclohexylglycine monohydrochloride is obtained as an off-white solid, mp1.
31-145 °, Rf = 0.36 (silica gel, n-butanol / acetic acid / water = 4: 1: 1). Elemental analysis (as C ₂₈ H ₃₅ N ₃ O ₅ · HCl · 0.7H ₂ O) Calculated value: C, 61.95, H, 6.95, N, 7.74, Cl,
6.53 Found: C, 61.95, H, 6.74, N, 7.71, Cl,
6.23

Example 3 (S) -7-[[[(±) -3- (benzoylamino) -2-oxo-4-phenylbutyl] methylamino] acetyl]-
Preparation of 1,4-dithia-7-azaspiro [4.4] nonane-8-carboxylic acid methyl ester :-( ±) -prepared as described in Example 2 (b) in distilled tetrahydrofuran (50 ml). To a solution of N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycine monohydrochloride (1.0 g, 2.5 mmol),
(S) -1,4-Dithia-7-azaspiro [4.4] nonane-
8-Carboxylic acid methyl ester monohydrochloride (0.66
g, 2.5 mmol), dicyclohexylcarbodiimide
Add (0.54 g, 2.5 mmol), hydrocylobenzobenzotriazole hydrate (0.39 g, 2.5 mmol) and diisopropylethylamine (0.9 ml, 5 mmol). The reaction mixture is stirred overnight, the precipitated dicyclohexylurea is filtered off and the filtrate is concentrated. The residue is ethyl acetate (100 ml)
, Saturated sodium bicarbonate (twice) and water (twice)
Wash with water, dry (Na ₂ SO ₄ ) and concentrate to a yellow oily residue (1.3 g). By flash chromatography (Merck silica gel, 25% ethyl acetate / methylene chloride, 1% methanol / methylene chloride) 0.53 g of (S) -7-
[[[(±) -3- (benzoylamino) -2-oxo-4-
Phenylbutyl] methylamino] acetyl] -1,4-dithia-azaspiro [4.4] nonane-8-carboxylic acid methyl ester is obtained as a white foam. mp 60-62 °, Rf
= 0.52 (silica gel, 5% methanol / methylene chloride). Elemental analysis (as C ₂₈ H ₃₃ N ₃ O ₅ S ₂ · 0.33H ₂ O) Calculated: C, 59.87, H, 6.04, N, 7.48, S,
11.42 Found: C, 59.87, H, 5.94, N, 7.56, S,
11.36

Example 4 (S) -7-[[[(±) -3- (benzoylamino) -2-oxo-4-phenylbutyl] methylamino] acetyl]-
Preparation of 1,4-dithia-7-azaspiro [4.4] nonane-8-carboxylic acid methyl ester monohydrochloride: -Methyl ester product of Example 3 (0.26 g, 0.46
(mmol) was treated with 2N hydrochloric acid / acetic acid until homogeneous (2 minutes), concentrated under reduced pressure, and the oily residue was triturated with ether (twice) to give 0.26 g of (S) -7-[[ [(±) -3- (benzoylamino) -2-oxo-4-phenylbutyl] methylamino] acetyl] -1,4-dithia-7-azaspiro [4.4] nonane-8-carboxylic acid methyl ester The monohydrochloride salt is obtained as a white solid. mp 79 to 85 °, Rf =
0.53 (silica gel, 5% methanol / methylene chloride). Elemental analysis (as C ₂₈ H ₃₃ N ₃ O ₅ S ₂ · HCl · 0.56H ₂ O) Calculated value: C, 55.84, H, 5.88, N, 6.98, S,
10.64, Cl, 5.88 Found: C, 55.84, H, 5.95, N, 6.78, S,
10.43, Cl, 5.66

Example 5 (4S) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -4-
Production of (4-fluorophenoxy) -L-proline methyl ester monohydrochloride: -a) Production of (4S) -4- (fluorophenoxy) -L-proline methyl ester monohydrochloride:-In methanol (4S) -4- (fluorophenoxy) -L
Thionyl chloride (8.09 ml, 11 mmol) was added to a suspension of proline (2.5 g, 11 mmol) at −30 ° under an argon atmosphere. The reaction mixture was stirred at -20 ° for 2 hours,
It is then stirred at room temperature for 16 hours. The solvent was removed under reduced pressure, the residue redissolved in methylene chloride (150 ml) and washed with 1N sodium carbonate (2 times) and water (2 times). Dry (MgSO ₄ )
Then, excess hydrochloric acid / methanol is added and the solvent is removed under reduced pressure. Ether was added to give a light brown solid (2.6g).
Recrystallize from methanol / ether to give 1.49 g (4
S) -4- (Fluorophenoxy) -L-proline methyl ester monohydrochloride is obtained as a light brown solid. mp14
7 to 148 °, [α] ²⁰ D = + 6.96 ° (C = 1.5
5, methanol).

B) (4S) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -4- (4-fluorophenoxy) -L-proline. Preparation of methyl ester :-( ±) -N- [3- (benzoylamino) prepared as described in Example 2 (b) in distilled tetrahydrofuran (20 ml).
2-oxo-4-phenylbutyl] -N-methylglycine monohydrochloride (1.17 g, 3 mmol) in a solution of (4
S) -4- (fluorophenoxy) -L-proline methyl ester monohydrochloride (0.82 g, 3 mmol), hydroxybenzotriazole hydrate (0.46 g, 3 mmol) and dicyclohexylcarbodiimide (0 .62g, 3m
Mol). The reaction mixture is stirred overnight, the precipitated dicyclohexylurea is filtered off and the filtrate is concentrated. The residue was dissolved in ethyl acetate (50 ml) and saturated sodium bicarbonate
Wash with (twice) and water (twice), dry (Na ₂ SO ₄ ) and concentrate to an oily residue (1.1 g). By flash chromatography (200 g Merck silica gel 60, 3% methanol / chloroform) 0.15 g (4S) -1-
[N- [3- (Benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -4- (4-fluorophenoxy) -L-proline methyl ester is obtained as a foam.

C) (4S) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -4- (4-fluorophenoxy) -2-proline. Preparation of methyl ester monohydrochloride: -Methyl ester product of (b) above (0.15 g, 0.26 m
(Mol) was treated with 2N hydrochloric acid / acetic acid until homogenous (2 minutes), concentrated under reduced pressure, and the oily residue was triturated with ether.
(2 times) to give 0.14 g of (4S) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-
Methylglycyl] -4- (4-fluorophenoxy) -L
-Proline methyl ester monohydrochloride is obtained as an off-white solid. mp105-125 °, Rf = 0.27
(Silica gel, 5% methanol / chloroform). Elemental analysis (as C ₃₂ H ₃₄ FO ₆ · HCl) Calculated value: C, 62.79, H, 5.76, N, 6.86, F,
3.10, Cl, 5.79 Found: C, 62.78, H, 5.73, N, 6.87, F,
2.83, Cl, 5.33

Example 6 (4S) -1- [N-[(S) -3- (benzoylamino) -2]
-Oxo-4-phenylbutyl] -L-alanyl] -4-
Preparation of (4-fluorophenoxy) -L-proline monohydrochloride: -a) (S) -N- [3- (benzoylamino) -2-oxo-
Preparation of 4-phenylbutyl] -L-alanine.1,1-dimethylethyl ester: (S) -N- [3-chloro-2-oxo-1- (phenylmethyl) propyl in dimethylformamide (80 ml) ] To a stirred solution of benzamide (10.0 g, 33.1 mmol),
L-alanine 1,1-dimethylethyl ester hydrochloride (6.0 g, 33.1 mmol), sodium bicarbonate (6.
1 g, 72 mmol) and sodium iodide (4.9 g, 3
3.1 mmol) is added. The resulting solution is stirred at room temperature overnight, poured into ether and washed with water (twice) and 10% sodium bicarbonate. The ether solution is extracted with 1N-hydrochloric acid (3 times), the combined extracts are made basic with solid sodium bicarbonate and extracted with ethyl acetate (4 times). The organic extracts were combined, dried (MgSO _4), to give a pale yellow solid 8.9g and concentrated. Part of this material was recrystallized from ethyl acetate,
(S) -N- [3- (benzoylamino) -2-oxo-4
-Phenylbutyl] -L-alanine.1,1-dimethylethyl ester is obtained as a white solid. mp106.5 ~ 11
0 °.

B) (S) -N- [3- (benzoylamino)-
Preparation of 2-oxo-4-phenylbutyl] -L-alanine monohydrochloride: -1.4 N-hydrochloric acid / acetic acid (39 ml) ester product of (a) above (2.95 g, 5.4 mmol) The solution of 1) is stirred at room temperature for 2 hours. The resulting white precipitate was collected, rinsed with ether and dried to give 2.27 g of (S) -N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanine monohydrochloride. Get the salt. mp 208-209 ° (decomposition), [α] D = -71 ° (C = 0.38%, methanol).
Rf = 0.51 (silica gel, chloroform / methanol / acetic acid = 4: 1: 1). Elemental analysis _{(C 20 H 22 N 2 O} 4 · HCl ) Calculated value: C, 61.64, H, 5.93 , N, 7.17, Cl,
9.07 Found: C, 61.33, H, 5.97, N, 7.17, Cl,
8.79

C) (S) -N- [N- (benzoylamino)-
Preparation of 2-oxo-4-phenylbutyl] -N-[(phenylmethoxy) carbonyl] -L-alanine :-( S) -N- [3- (benzoylamino) -2-oxo-4
-Phenylbutyl] -L-alanine monohydrochloride (2.0
g, 5.1 mmol), benzyl chloroformate (730 μl,
To a mixture of 5.1 mmol), water (7 ml) and dioxane (7 ml) was added triethylamine (2.1 ml, 15 mmol) 25
Add in °. The resulting mixture was stirred at 25 ° for 3 hours,
Then pour into 5% aqueous sodium bicarbonate and wash with ether. The aqueous layer was acidified (HCl) and extracted with ethyl acetate (3
Times) The extracts were dried (MgSO _4), to give a colorless oil and concentrated. Trituration with ether gives a white granular solid which is collected and discarded. The mother liquor was concentrated under reduced pressure and 1.75 g of (S) -N- [3- (benzoylamino) -2-
Oxo-4-phenylbutyl] -N-[(phenylmethoxy) carbonyl] -L-alanine is obtained as a white glass.

D) (4S) -1- [N-[(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-
[(Phenylmethoxy) carbonyl] -L-alanyl] -4
Preparation of-(4-fluorophenoxy) -L-proline phenylmethyl ester :-( S) -N in tetrahydrofuran (7 ml)
-[N- (benzoylamino) -2-oxo-4-phenylbutyl] -N-[(phenylmethoxy) carbonyl] -L
-Alanyl (300 mg, 0.62 mmol), (4S) -4-
(4-Fluorophenoxy) -L-proline phenylmethyl ester p-toluenesulfonate (300 mg,
0.62 mmol), triethylamine (90 μl, 0.62
mmole), dicyclohexylcarbodiimide (130 mg,
A mixture of 0.62 mmol) and hydroxybenzotriazole hydrate (90 mg, 0.62 mmol) is stirred at 25 ° for 20 hours. Then the mixture is filtered and diluted with ethyl acetate. The resulting solution was washed successively with 1N hydrochloric acid and 10% aqueous sodium bicarbonate solution, dried (MgSO ₄ ),
Filter and concentrate to 500 mg of (4S) -1- [N-[(S)-
3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-[(phenylmethoxy) carbonyl] -L-alanyl] -4- (4-fluorophenoxy) -L-proline phenylmethyl ester Obtained as a yellow oil.

E) (4S) -1- [N-[(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-
Preparation of alanyl] -4- (4-fluorophenoxy) -L-proline monohydrochloride: -Ester product of (d) above (500 mg, 0.6 mmol), palladium on carbon catalyst (10%, 100 mg) , Absolute ethanol (15 ml) and 1.0 N aqueous hydrochloric acid (800 μl, 0.
8 mmol) of the mixture is hydrogenated at 1 atm and 25 ° for 17 hours, then filtered and concentrated. The residue is HP-2
At 0, [0.01 N-aqueous hydrochloric acid / methanol = 9: 1]
To [0.01 N-aqueous hydrochloric acid / methanol = 1: 1]. Predetermined product (TL
Fractions containing C) are collected and concentrated. Dissolve the residue in the minimum amount of methanol. Ether was added to form a white precipitate, which was collected and dried under reduced pressure to give 200 mg of (4S) -1.
-[N-[(S) -3- (benzoylamino) -2-oxo-
4-Phenylbutyl] -L-alanyl] -4- (4-fluorophenoxy) -L-proline monohydrochloride is obtained.
mp152-153 ° (decomposition), [α] ²⁵ D = −50 ° (C
= 0.5, methanol). Rf = 0.75 (silica gel,
Chloroform / methanol / acetic acid = 4: 1: 1). Elemental analysis (as C ₃₁ H ₃₂ FN ₃ O ₆ · HCl · 1.5H ₂ O) Calculated value: C, 59.57, H, 5.80, N, 6.72, Cl,
5.67 Found: C, 59.68, H, 5.56, N, 6.67, Cl,
5.99

Example 7 [1 (S), 4R] -1- [N-3- (benzoylamino) -2
-Oxo-4-phenylbutyl] -L-alanyl] -4-
Preparation of phenyl-L-proline monohydrochloride: -a) (S) -N- [N- (benzoylamino) -2-oxo-
Preparation of 4-phenylbutyl] -N-[(phenylmethoxy) carbonyl] -L-alanyl succinimide ester:
-(S) -N- [N- (benzoylamino) -2, prepared as described in Example 6 (c), in tetrahydrofuran (5 ml).
-Oxo-4-phenylbutyl] -N-[(phenylmethoxy) carbonyl] -L-alanine (800 mg, 1.6 mmol), dicyclohexylcarbodiimide (340 mg, 1.
6 mmol) and N-hydroxysuccinimide (190 mmol)
g, 1.6 mmol) is stirred at 25 ° for 18 hours. After this time, filter and concentrate to give 950 mg of (S) -N-
[N- (benzoylamino) -2-oxo-4-phenylbutyl] -N-[(phenylmethoxy) carbonyl] -L-
Alanyl succinimide ester is obtained.

[1 (S), 4R] -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-
[(Phenylmethoxy) carbonyl] -L-alanyl] -4
Preparation of phenyl-L-proline: (S) -N- [N- (benzoylamino) -2-oxo-4-phenylbutyl] -N in dimethylformamide (5 ml)
A solution of-[(phenylmethoxy) carbonyl] -L-alanine succinimide ester (950 mg, 1.6 mmol) was added to (4R) -4-phenyl-L-proline hydrochloride (3
75 mg, 1.7 mmol) and triethylamine (40 μ)
l, 3.2 mmol) is added. The resulting mixture is stirred at 25 ° for 24 hours, then poured into excess 1N hydrochloric acid and extracted with ethyl acetate (3 times). The combined extracts were dried (MgSO ₄₎
, Filtered and concentrated to give 1.1 g of [1 (S), 4R] -1-
[N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-[(phenylmethoxy) carbonyl]-
L-alanyl] -4-phenyl-L-proline is obtained.

C) [1 (S), 4R] -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L
-Production of alanyl] -4-phenyl-L-proline monohydrochloride: -Product of (b) above (1.0 g, 1.5 mmol), ethanol
(20 ml), water (5 ml), 1.0 N-hydrochloric acid (1.5 ml, 1.
5 mmol) and palladium / carbon catalyst (10%, 100 mmol)
g) is hydrogenated at 1 atm and 25 ° for 18 hours, then filtered and concentrated. The residue is [0.0] on HP-20.
Chromatography using a linear gradient of 1N-aqueous hydrochloric acid: methanol = 4: 60 to 10:90]. Fractions containing the desired product (TLC) are collected and concentrated. Dissolve the residue in the minimum amount of methanol. Ether was added and the resulting white precipitate was collected and dried to give 300 mg of [1 (S), 4R].
-1- [N- [3- (benzoylamino) -2-oxo-4
-Phenylbutyl] -L-alanyl] -4-phenyl-L
-Proline monohydrochloride is obtained. mp160 ~ 162 °
(Decomposition), [α] ²⁵ D = −57 ° (C = 1.5, methanol), Rf = 0.8 (silica gel, chloroform / methanol / acetic acid = 4: 1: 1). Elemental analysis (as C ₃₁ H ₃₃ N ₃ O ₅ · HCl · 1.35H ₂ O) Calculated value: C, 63.27, H, 6.29, N, 7.14, Cl,
6.02 found: C, 63.27, H, 6.17, N, 7.19, Cl,
5.97

Example 8 [1 (s), 4s] -1- [N- [3- (benzoylamino) -2]
-Oxo-4-phenylbutyl] -L-alanyl] -4-
Production of phenyl-L-proline monohydrochloride: In the same manner as in Example 7 except that (4s) -4-phenyl-L-proline.hydrochloride was used in Example 7 (b), [1
(s), 4s] -1- [N- [3- (Benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl-4-phenyl-L-proline monohydrochloride is obtained. mp138-
143 °, [α] D = -61 ° (c = 0.3%, methanol). Rf = 0.84 (silica gel; chloroform / methanol / acetic acid = 4: 1: 1). Elemental analysis (as C ₃₁ H ₃₃ N ₃ O ₅ · HCl · 2.13H ₂ O) Calculated value: C, 61.80, H, 6.35, N, 6.98, Cl,
5.88 Found: C, 61.80, H, 6.06, N, 7.05, Cl,
5.65

Example 9 [1 (s), 4R] -1- [N- [3- (benzoylamino) -2]
-Oxo-4-phenylbutyl] -L-alanyl] -4-
Preparation of cyclohexyl-L-proline monohydrochloride: In Example 7 (b), (4R) -4-cyclohexyl-
[1 (s), 4R] -1- [N- [3- (benzoylamino)) was prepared in the same manner as in Example 7 except that L-proline hydrochloride was used.
-2-Oxo-4-phenylbutyl] -L-alanyl]-
4-Cyclohexyl-L-proline monohydrochloride is obtained. mp 140-154 ° (decomposition), [α] D = -83 ° (c
= 0.36%, methanol). Rf = 0.84 (silica gel, chloroform / methanol / acetic acid 4: 1: 1). Elemental analysis (as C ₃₁ H ₃₉ N ₃ O ₅ .HCl.0.79H ₂ O) Calculated value: C, 63.71, H, 7.17, N, 7.19, Cl,
6.06 Found: C, 63.71, H, 7.21, N, 7.05, Cl,
5.82

Example 10 [1 (s), 4s] -1- [N- [3- (benzoylamino) -2]
-Oxo-4-phenylbutyl] -L-alanyl] -4-
Preparation of cyclohexyl-L-proline monohydrochloride: -in Example 7 (b), (4s) -4-cyclohexyl-L
-[1 (s), 4s] -1- [N- [3- (benzoylamino) -in the same manner as in Example 7 except that proline hydrochloride was used.
2-oxo-4-phenylbutyl] -L-alanyl] -4
-Cyclohexyl-L-proline monohydrochloride is obtained.
mp139-141 ° (decomposition), [α] D = -80 ° (c =
0.2%, methanol). Rf = 0.83 (silica gel,
Chloroform / methanol / acetic acid = 4: 1: 1). Elemental analysis (as C ₃₁ H ₃₉ N ₃ O ₅ · HCl · 1.54H ₂ O) Calculated value: C, 62.28, H, 7.26, N, 7.03, Cl,
5.93 Found: C, 62.28, H, 7.01, N, 7.02, Cl,
6.16

Example 11 (s) -7-[(s) -2-[[3- (benzoylamino) -2-]
Preparation of oxo-4-phenylbutyl] amino] -1-oxopropyl] -1,4-dithia-7-azaspiro [4.4] nonane-8-carboxylic acid monohydrochloride: -Example 7 (b) In place of the L-proline reactant
(s) -1,4-dithia-7-azaspiro [4.4] nonane-
(S) -7-[(s) -2-[[3- (benzoylamino)) in the same manner as in Example 7 except that 8-carboxylic acid / hydrochloride was used.
-2-oxo-4-phenylbutyl] amino] -1-oxopropyl] -1,4-dithia-7-azaspiro [4.4]
Nonane-8-carboxylic acid monohydrochloride is obtained. mp17
0-172 °, [α] ²⁵ D = -26 ° (c = 1.4%, methanol). Rf = 0.78 (silica gel, chloroform /
Methanol / acetic acid = 6: 1: 1). Elemental analysis (as C ₂₇ H ₃₁ N ₃ O ₅ S ₂ · HCl · 0.77H ₂ O) Calculated value: C, 54.77, H, 5.71, N, 7.10, S,
10.83, Cl, 5.99 Found: C, 54.77, H, 5.70, N, 6.94, S,
10.82, Cl, 6.07

Example 12 [1 (s), 5s] -1- [N- [3- (benzoylamino) -2]
-Oxo-4-phenylbutyl] -L-alanyl] -4,
5-dihydro-3-phenyl-1H-pyrazole-5-
Preparation of Carboxylic Acid Monohydrochloride: In Example 7 (b), Instead of the L-Proline Reactant
The title compound is obtained in the same manner as in Example 7 except that (s) -4,5-dihydro-3-phenyl-1H-pyrazole-5-carboxylic acid is used.

Example 13 (s) -2- [N-[(s) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -1,2,
3,4-Tetrahydro-3-isoquinolinecarboxylic acid
Preparation of monohydrochloride: -In Example 7 (b), (s) -instead of the proline reactant.
(S) -in the same manner as in Example 7 except that 1,2,3,4-tetrahydro-3-isoquinolinecarboxylic acid hydrochloride was used.
2- [N-[(s) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -1,2,3,4
-Tetrahydro-3-isoquinolinecarboxylic acid monohydrochloride is obtained.

Example 14 1- [N-[(s) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -2- (2-hydroxyphenyl) -4 (R ) -Preparation of thiazolidinecarboxylic acid monohydrochloride: -In Example 7 (b), instead of the proline reactant, 2-
After removing the hydroxy protecting group, 1- [N-[(s) -3 was prepared in the same manner as in Example 7 except that [(2-phenylmethoxy) phenyl] -4 (R) -thiazolidinecarboxylic acid / hydrochloride was used.
-(Benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -2- (2-hydroxyphenyl)-
4 (R) -thiazolidinecarboxylic acid monohydrochloride is obtained.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI Technical display location C07D 231/14 6701-4C 307/54 7729-4C 333/24 7729-4C 495/10 7329-4C // C07D 205/04 7019-4C 207/16 7019-4C 207/22 7019-4C 277/06 7019-4C (C07D 495/10 207: 00 339: 00) (72) Inventor Eritsk M. Gordon United States New Jersey, Pennington, Raining Avenue No. 126

Claims (5)

[Claims] 1. The formula: A compound represented by the formula: wherein R is hydrogen, Prot, lower alkyl, cycloalkyl, R ₁ is hydrogen, lower alkyl, halo-substituted (lower) alkyl, embedded image (However, when R is a group other than hydrogen, R ₁ is hydrogen); Pro
t is an easily removable group; R ₂ is R ₃ is hydrogen, lower alkyl, [Chemical 6] R ₁₄ is hydrogen, lower alkyl having 1 to 4 carbons, 1 to 4 carbons
4 lower alkoxy, C1-4 lower alkylthio, chloro, bromo, fluoro, trifluoromethyl or hydroxy; m is 0, 1, 2, 3 or 4; p is 1, 2
Or 3 (provided that R ₁₄ is hydrogen, methyl, methoxy, chloro or fluoro, p is an integer greater than 1); r
Represents an integer of 1 to 4]. 2. R ₂ is R ₃ is The compound of claim 1 wherein m is 0, 1 or 2; R ₁₄ is hydrogen, methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy; R is methyl; R ₁ is hydrogen. 3. The compound according to claim 2, wherein R ₂ is phenyl and R ₃ is benzyl. 4. R ₂ is R ₃ is The compound according to claim 1, wherein m is 0, 1 or 2; R ₁₄ is hydrogen, methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy; a group in which R is easily removable; R ₁ is methyl. 5. R ₂ is phenyl; R ₃ is benzyl; R is The compound according to claim 4, which is