JPH09124691A - Peptide compound and medicinal composition containing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new peptide compound, having a specific amino acid sequence, capable of manifesting excellent inhibiting actions of the chymase group on mammals including humans and useful for prevention and treatment, etc., of diseases caused by angiotensin II and diseases caused by the chymases. SOLUTION: This new peptide (salt) is represented by formula I X is represented by formula II [ring B is an aromatic ring; Z1 and Z2 are each H, OH, a group represented COR<8> (R<8> is OH, an alkoxy, amino, etc.)], etc.; R1 is a (substituted)alkyl, an alkoxy, an aryl, a heteroacyl, etc.; R2 and R3 are each H, a (substituted)alkyl, a cycloalkyl, an aryl, etc.; R4 and R5 are each H, an alkyl, etc.; Y is a cycloalkyl, an aryl or a heteroaryl; (m) is 0 or 1-3; A is carbonyl or sulfonyl} and is useful for prevention, treatment, etc., of diseases caused by angiotensin II, diseases, etc., caused by chymases. The compound is obtained by reacting a compound, represented by formula IV and induced from an aldehyde represented by formula III (Rp is a protecting group) with a dipeptide represented by formula V, then amidating the C-terminal and further acylating the N-terminal thereof.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel peptide compound, a pharmacologically acceptable salt thereof, a pharmaceutical composition thereof and a pharmaceutical use thereof. In particular, the present invention relates to a peptide compound useful in pharmacology, diagnosis, and prevention / treatment of diseases, a pharmacologically acceptable salt thereof, and a pharmaceutical composition containing the same. The present invention also relates to intermediates required for the synthesis of the above peptide compounds.

[0002]

2. Description of the Related Art Angiotensin II has physiological activities such as blood pressure increase by strongly contracting blood vessels and stimulation of secretion of aldosterone having a sodium retention effect from adrenal cortex, hypertension, cardiac hypertrophy, myocardial infarction, It is considered to be a causative agent or risk factor for diseases such as arteriosclerosis, diabetic and non-diabetic renal diseases, and vascular restenosis after PTCA (percutaneous transluminal coronary angioplasty). This angiotensin II is produced by cleaving 2 amino acid residues from angiotensin I, which is a peptide consisting of 10 amino acids existing in the living body, and angiotensin converting enzyme (ACE) is involved in the cleavage. It is known that many ACE inhibitors have been developed as prophylactic / therapeutic agents for the above diseases.

[0003] In recent years, the action of chymase groups including human heart chymase, human mast cell chymase and human skin chymase, which are one of the subfamilies of serine proteases, has been receiving attention. Chymase has been shown to be involved in the process of angiotensin II production that is not based on ACE in the above-mentioned conversion of angiotensin I to angiotensin II (Okunishi et al., Jp.
n. J. Pharmacol. 1993, 62, p207 etc.) and many other physiologically active substances such as extracellular matrix, cytokines, substance P, VIP (Vasoactive Intestinal Polypeptide) and apoprotein B as substrates. It is also known to be involved in the activation of other proteases such as collagenase (Miyazaki
Et al., History of Medicine 1995, 172, p559).

[0004] Therefore, chymase inhibitors are expected to be inhibitors of angiotensin II action by suppressing the production of angiotensin II independent of ACE, and as preventive and therapeutic agents for various diseases caused by chymase. And a patent application for a chymase inhibitor based on these ideas has already been filed (WO93 / 25574).

By the way, the above-mentioned patent WO93 / 25574 discloses an inhibitor compound of a chymase group including human heart chymase, which has a structure similar to that of the compound (I) of the present invention. But,
These compounds also inhibit other enzymes such as chymotrypsin, which has a substrate similar to that of chymase, and have insufficient selectivity for chymase group including human heart chymase, and chymase inhibitory activity is not always satisfactory. Absent.

[0006]

DISCLOSURE OF THE INVENTION An object of the present invention is to provide a novel compound having a chymase inhibitory activity excellent in selectivity and / or a high chymase inhibitory activity, and a pharmaceutical composition containing the same. Is.

[0007]

Means for Solving the Problems As a result of intensive studies aimed at achieving the above-mentioned object, the present inventors have synthesized various novel compounds, and other enzymes such as chymotrypsin whose substrate is close to that of chymase. The present invention has been made to find a compound which inhibits chymase group including human cardiac chymase with high selectivity and / or a compound having high chymase inhibitory activity without inhibiting the enzyme.

That is, the present invention provides a compound of the formula (I)

[0009]

Embedded image

[Wherein X is

[0011]

Embedded image

Or

[0013]

Embedded image

(Wherein Z ₁ and Z ₂ are the same or different and are hydrogen, a hydroxyl group, a group represented by --COR ₈ or --COR
_An alkyl optionally substituted with a group represented by ₈ ,
Or a group represented by —SO ₂ R ₈ , R ₆ and R
₇ are the same or different and each is hydrogen, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkyl substituted with a group represented by the group or -COR ₈ represented by -COR _8, Alkenyl, alkynyl, heterocycle or heterocycle alkyl, provided that
When R ₆ and R ₇ do not represent hydrogen at the same time, and either or both of R ₆ and R ₇ are a heterocycle substituted with a group represented by —COR ₈ , the heteroatom of the heterocycle is R _6, not bonded to the carbon atom bonded to the R _7, or R ₆ and R ₇ together -COR
_{4 to 8} carbon atoms which may be substituted with a group represented by ₈
6 methylene chain may be shown, and B ring is an aromatic ring)
Represents a group represented by, R ₁ is alkyl optionally substituted with a group represented by —COR ₈ , cycloalkyl, cycloalkylalkyl, alkoxy, aryl,
Arylalkyl, heteroaryl, heterocycle or heterocycle alkyl is shown, and R ₂ and R ₃ are the same or different and are each hydrogen, or alkyl optionally substituted with a group represented by —COR ₈ , cycloalkyl,
Represents cycloalkylalkyl, aryl or arylalkyl, or R ₂ and R ₃ together are --CO
C 4 optionally substituted with a group represented by R ₈
~ 6 methylene chain may be represented, R ₄ represents hydrogen or alkyl, R ₅ represents hydrogen or alkyl, or X is

[0015]

Embedded image

In the case of, the number of carbon atoms is 2 to 4 together with R _6.
R ₈ may be a hydroxyl group, alkoxy, amino, alkylamino, dialkylamino, aryloxy or arylalkoxy, Y is cycloalkyl, aryl or heteroaryl, m
Represents an integer of 0 or 1 to 3, and A represents carbonyl or sulfonyl] (hereinafter, also referred to as compound (I)) or a pharmaceutically acceptable salt thereof.

The invention also relates to formula (II) useful for the synthesis of compounds of formula (I)

[0018]

Embedded image

[Wherein X is

[0020]

Embedded image

Or

[0022]

Embedded image

(In the formula, Z ₁ and Z ₂ are the same or different and each is hydrogen, a hydroxyl group, a group represented by --COR ₈ , or --COR
_An alkyl optionally substituted with a group represented by ₈ ,
Or a group represented by —SO ₂ R ₈ , R ₆ and R
₇ are the same or different and each is hydrogen, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, alkyl substituted with a group represented by the group or -COR ₈ represented by -COR _8, Alkenyl, alkynyl, heterocycle or heterocycle alkyl, provided that
When R ₆ and R ₇ do not represent hydrogen at the same time, and either or both of R ₆ and R ₇ are a heterocycle substituted with a group represented by —COR ₈ , the heteroatom of the heterocycle is R _6, not bonded to the carbon atom bonded to the R _7, or R ₆ and R ₇ together -COR
_{4 to 8} carbon atoms which may be substituted with a group represented by ₈
6 methylene chain may be shown, and B ring is an aromatic ring)
And R ₂ and R ₃ are the same or different and each is hydrogen, or an alkyl, cycloalkyl, cycloalkylalkyl, aryl or arylalkyl which may be substituted with a group represented by —COR _8. Or R ₂ and R ₃ together may represent a methylene chain having 4 to 6 carbon atoms, which may be substituted with a group represented by —COR ₈ , and R ₄ represents hydrogen or alkyl. R ₅ is hydrogen or alkyl, or X is

[0024]

Embedded image

In the case of, the number of carbon atoms is 2 to 4 together with R _6.
R ₈ represents a hydroxyl group, alkoxy, amino, alkylamino, dialkylamino, aryloxy or arylalkoxy, Rp represents an amino-protecting group, and Y represents cycloalkyl, aryl or hetero. Which represents aryl, m represents 0 or an integer of 1 to 3, and A represents carbonyl or sulfonyl] or a pharmacologically acceptable salt thereof.

Further, the present invention relates to a pharmaceutical composition containing the compound (I) or a pharmacologically acceptable salt thereof, and a pharmaceutical use thereof, especially a chymase inhibitor.

[0027]

BEST MODE FOR CARRYING OUT THE INVENTION

The symbols used in this specification are explained below. The alkyl in R _{1 to} R ₇ , Z ₁ and Z ₂ preferably has 1 to 6 carbon atoms and may be linear or branched, for example, methyl, ethyl, n-propyl,
Isopropyl, n-butyl, isobutyl, sec-butyl,
tert-butyl, n-pentyl, isopentyl, n-hexyl and the like can be mentioned.

Cycloalkyl in R _{1 to} R ₃ , R ₆ , R ₇ and Y preferably has 3 to 7 carbon atoms and includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.

The cycloalkylalkyl in R _{1 to} R ₃ , R ₆ and R ₇ has the same cycloalkyl moiety as described above, and the alkyl moiety preferably has 1 to 1 carbon atoms.
3 may be linear or branched, and examples thereof include cyclopropylmethyl, 2-cyclobutylethyl, 3-cyclopentylpropyl, cyclohexylmethyl, 2-cyclohexylethyl, cycloheptylmethyl and the like.

Alkoxy for R ₁ and R ₈ is
The alkyl part is the same as above, for example, methoxy,
Ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy,
Examples include n-pentoxy, isopetoxy, n-hexoxy and the like.

Aryl in R _{1 to} R ₃ , R ₆ , R ₇ and Y is preferably a phenyl, naphthyl or ortho-fused bicyclic group having 8 to 10 ring atoms,
Examples thereof include those in which at least one ring is an aromatic ring (for example, indenyl, etc.).

The arylalkyl in R _{1 to} R ₃ , R ₆ and R ₇ has the same aryl part as described above, and the alkyl part preferably has 1 to 3 carbon atoms and is linear or branched. However, for example, benzyl, phenethyl,
3-phenylpropyl, 1-naphthylmethyl, 2-naphthylmethyl, 2- (1-naphthyl) ethyl, 2- (2-
Naphthyl) ethyl, 3- (1-naphthyl) propyl, 3
-(2-naphthyl) propyl and the like can be mentioned.

The heteroaryl in R ₁ , R ₆ , R ₇ and Y is preferably a 5- or 6-membered ring group having carbon and 1 to 4 heteroatoms (oxygen, sulfur or nitrogen),
Or an ortho-fused bicyclic heteroaryl having 8 to 10 ring atoms derived therefrom, in particular a benz derivative, or a derivative thereof fused with a propenylene, trimethylene or tetramethylene group, and a stable N-- Examples thereof include oxides. For example, pyrrolyl, furyl, thienyl, oxazolyl, isoxazolyl, imidazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl, tetrazolyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl, pyridyl, pyranyl, pyrazinyl, pyrimidinyl, pyridazinyl, 1,2,4-triazinyl, 1,2,3-triazinyl, 1,3,5-triazinyl, 1,2,5-oxathiazinyl, 1,2,6-oxathiazinyl, benzoxazolyl, benzothiazolyl, benzimidazolyl, thianaphthenyl, Isothianaphthenyl, benzofuranyl,
Examples thereof include isobenzofuranyl, chromenil, isoindolyl, indolyl, indazolyl, isoquinolyl, quinolyl, phthalazinyl, quinoxalinyl, quinazolinyl, cinnolinyl and benzoxazinyl.

The heteroarylalkyl in R ₆ and R ₇ has the same heteroaryl part as described above, and the alkyl part preferably has 1 to 3 carbon atoms and may be linear or branched. 2-pyrrolylmethyl, 2
-Pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, 2-thienylmethyl, 2- (2-pyridyl) ethyl, 2- (3-pyridyl) ethyl, 2- (4-pyridyl) ethyl, 3- (2 -Pyrrolyl) propyl and the like.

The heterocycle represented by R ₁ , R ₆ and R ₇ means carbon and 1 to 4 heteroatoms (oxygen,
A 4- to 6-membered ring group having (sulfur or nitrogen), for example, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, oxothiomorpholinyl, dioxothiomorpholinyl, tetrahydropyranyl, dioxacyclohexyl, etc. Can be mentioned.

The heterocycle alkyl in R ₁ , R ₆ and R ₇ has the same heterocycle part as described above, and the alkyl part preferably has 1 to 3 carbon atoms and may be linear or branched. Well, for example azetidinoethyl,
Examples thereof include pyrrolidinopropyl, piperidinomethyl, piperazinoethyl, morpholinopropyl, thiomorpholinomethyl, dioxacyclohexylmethyl and the like.

Alkenyl for R ₆ and R ₇ is
It preferably has 3 to 6 carbon atoms and may be linear or branched, and examples thereof include 2-propenyl, 3-butenyl, 4-pentenyl and 5-hexenyl.

Alkynyl for R ₆ and R ₇ is
It preferably has 3 to 6 carbon atoms and may be linear or branched, and examples thereof include 2-propynyl, 3-butynyl, 4-pentynyl, and 5-hexynyl.

Alkylamino in R ₈ is preferably an alkyl moiety having 1 to 6 carbon atoms, which may be linear or branched and includes, for example, methylamino, ethylamino, n.
-Propylamino, isopropylamino, n-butylamino, isobutylamino, sec-butylamino, tert-butylamino, n-pentylamino, isopentylamino, n-hexylamino and the like.

The dialkylamino in R ₈ is preferably an alkyl part having 1 to 6 carbon atoms and may be linear or branched, and examples thereof include dimethylamino, diethylamino, di-n-propylamino, diisopropylamino, Examples thereof include di-n-butylamino, diisobutylamino, disec-butylamino, di-n-pentylamino, di-isopentylamino, di-n-hexylamino and the like.

The aryloxy in R ₈ is preferably a phenyl, naphthyl, or ortho-fused bicyclic group having 8 to 10 ring atoms, at least one ring of which is an aromatic ring. Some examples (such as indenyl) and the like can be mentioned.

Arylalkoxy in R ₈ is preferably a phenyl, naphthyl, or ortho-fused bicyclic group having 8 to 10 ring atoms.
Examples thereof include those in which at least one ring is an aromatic ring (eg, indenyl, etc.), and the alkyl portion thereof preferably has 1 to 6 carbon atoms and may be linear or branched, for example, benzyloxy, 2 -Phenylethoxy, 3-phenylpropoxy, 1-naphthylmethoxy, 2-naphthylmethoxy, 2- (1-naphthyl) ethoxy, 2- (2-naphthyl) ethoxy and the like can be mentioned.

Examples of the B ring include those represented by the above-mentioned aryl and heteroaryl.

Examples of the amino group-protecting group for Rp include a carbamate-type protecting group, an acyl-type protecting group and an alkyl-type protecting group. Examples of the carbamate-type protecting group include a benzyloxycarbonyl group (Z) and a tertiary-type protecting group. Secondary butyloxycarbonyl group (Boc), isobonyloxycarbonyl group (Iboc), p-biphenylisopropyloxycarbonyl group (Bpoc), 9-fluorenylmethyloxycarbonyl group (Fmoc) and the like, and acyl-type protecting groups. Examples thereof include formyl group, p-toluenesulfonyl group (Tos) and the like, and examples of alkyl type protecting group include trityl group (Trt) and the like.

In the compound (I) of the present invention, X is preferably a compound of formula (I).

[0047]

Embedded image

(Wherein Z ₁ and Z ₂ are the same or different and are hydrogen, a hydroxyl group, a group represented by --COR ₈ or --COR
_An alkyl optionally substituted with a group represented by ₈ ,
Or a group represented by —SO ₂ R ₈ ), R ₁ is alkyl, R ₂ and R ₃ are the same or different and are hydrogen or a group represented by —COR _8. R ₄ represents hydrogen, R ₅ represents hydrogen or alkyl, R ₈ represents a hydroxyl group, alkoxy or arylalkoxy, and Y represents an optionally substituted alkyl.
Is aryl, m is 1, and A is carbonyl or sulfonyl.

Among the above compounds, at least one of Z ₁ and Z ₂ is a group represented by --COR ₈ or --CO.
Particularly preferred is a compound showing alkyl substituted by a group represented by R _8.

Further, among the compounds of the present invention, as a preferable compound, X in the formula (I) is

[0051]

Embedded image

[0052] (shown wherein hydrogen R _6, R ₇ are the same or different, aryl, arylalkyl, an alkyl substituted with a group represented by the group or -COR ₈ represented by -COR _8, where R ₆ and R ₇ do not represent hydrogen at the same time), R ₁ represents alkyl, and R ₂ and R ₃ are the same or different and are hydrogen or --CO.
R ₈ represents alkyl which may be substituted with a group represented by R ₈ , R ₄ represents hydrogen, R ₅ represents hydrogen or alkyl, or R ₆ together represents a methylene chain having 3 carbon atoms. R ₈ represents a hydroxyl group, alkoxy or arylalkoxy, Y represents aryl, and m is 1
And A is carbonyl or sulfonyl.

Of the above compounds, at least one of R ₆ and R ₇ is a group represented by —COR ₈ or —COR.
Especially preferred are compounds exhibiting alkyl substituted with a group represented by ₈ .

The present invention also includes all optically active isomers and diastereomeric mixtures based on the asymmetric carbon which compound (I) has. These can be separated individually by a method known per se. Compound (I) can also exist as a solvate (eg, ketone solvate, hydrate, etc.). Therefore, the present invention includes any of the above stereoisomers, optical isomers, solvates, and any mixture thereof.

When compound (I) is an acidic compound, pharmacologically acceptable salts thereof include alkali metal salts (eg salts with lithium, sodium, potassium etc.) and alkaline earth metal salts (eg calcium). , A salt with magnesium, etc.), an aluminum salt, an ammonium salt, a salt with an organic base (for example, a salt with triethylamine, morpholine, piperidine, triethanolamine, etc.) and the like. When compound (I) is a basic compound, its pharmacologically acceptable salts include inorganic acid addition salts (eg, salts with hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, etc.) , Organic acid addition salts (eg, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, formic acid, acetic acid, trifluoroacetic acid, oxalic acid, citric acid, malonic acid, fumaric acid, glutaric acid, adipic acid, maleic acid ,
Salts with tartaric acid, succinic acid, mandelic acid, malic acid, etc.),
Salts with amino acids (eg, salts with glutamic acid, aspartic acid, and the like) and the like can be mentioned.

Next, a method for producing the compound (I) of the present invention will be described. The main route for producing the compound (I) of the present invention is shown in the following scheme 1.

[0057]

Embedded image

(Step a) L- having an amino group protected by a protecting group Rp prepared by a method known per se.
Alternatively, using D-amino acid as a starting material, Fehrentz (Syn
The aldehyde compound (1) obtained according to the method of thesis 1983, 678) is synthesized. Next, the aldehyde compound (1)
Hallinan and Fried (Tetrahedron Lett. 1984, 25, 2
301) and Thairivongs et al. (J. Med. Chem. 1986, 29, 208).
0) by reacting with ethyl bromodifluoroacetate in the presence of zinc dust, or by Lang and Schaub (Tetrahedron
Lett. 1988, 29. 2943) and reacted with ethyl chlorodifluoroacetate in the presence of zinc dust, or according to the method of Hoover (US Pat. No. 4,855303) ethyl bromodifluoroacetate, zinc dust and The ester compound (2) is synthesized by reacting with titanium tetrachloride.

(Step b) protecting group R of ester compound (2)
The amine compound is obtained by deprotecting p by a method known per se (hydrogenation reaction, acid decomposition, or hydrolysis).
Combine (3).

(Step c) The amine compound (3) is subjected to a condensation reaction with a carboxylic acid represented by the formula (4) (a synthetic method will be described later) or a reactive derivative thereof to give the compound (5).
Are synthesized. The amounts of the compound (3) and the carboxylic acid (4) or its reactive derivative to be charged may be usually equimolar, but if necessary, one of them is used in an amount of 1.1 to 3 times that of the other. When the carboxylic acid (4) is used as it is, the reaction is carried out in the presence of a condensing agent. As the condensing agent, 2-chloro-
4,6-Dimethoxy-1,3,5-triazine, O-benzotriazol-1-yl-N, N, N, N'-tetramethyluronium hexafluorophosphate, benzotriazol-1-yl-oxy-tri Pyrrolidinophosphonium hexafluorophosphate, benzotriazol-1-yl-oxy-tris (dimethylamino)
Phosphonium hexafluorophosphate, N, N-dicyclohexylcarbodiimide, N, N-diisopropylcarbodiimide, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide methiozide, 1-ethyl-3- (3-dimethylaminopropyl) Carbodiimide hydrochloride, N-cyclohexyl-N '-(2-morpholinoethyl) carbodiimide meth-p-toluenesulfonate, etc. are used. As the carboxylic acid (4), those converted into a reactive derivative such as an acid anhydride, an active ester or an acid halide by a conventional method may be used. As the acid anhydride, for example, an anhydride with pivalic acid, an anhydride with carbonic acid isobutyl ester, or the like is used. As the active ester, for example, p-nitrophenyl ester, 2,4,5-trichlorophenyl ester, N-hydroxysuccinimide ester, N-hydroxyphthalimide ester or the like is used. As the acid halide, for example, carboxylic acid chloride, carboxylic acid bromide and the like are used. As the reaction solvent, in any case, N, N-dimethylformamide, N, N-dimethylacetamide, dimethyl sulfoxide, hexamethylphosphoric triamide, pyridine, dioxane, tetrahydrofuran, acetonitrile, chloroform, methylene chloride, dimethoxyethane. , Benzene, ethyl acetate, or a mixed solvent thereof is used. Preferred solvents include N, N-dimethylformamide, methylene chloride, tetrahydrofuran, acetonitrile. Usually, the reaction temperature is about 0-100 ° C, and the reaction time is several hours to 5 days. In the above reaction, N-methylmorpholine,
A basic reaction aid such as 4-dimethylaminopyridine or triethylamine can be used.

(Step d) by reacting the compound (5) with the amine compound (6) in a protic polar solvent, preferably ethanol or methanol, or
(5) is hydrolyzed by a conventional method under basic conditions such as sodium hydroxide and potassium hydroxide, and the obtained carboxylic acid alkali salt and amine compound (6) are used as a reaction auxiliary agent such as 1-hydroxybenzotriazole. In the presence of the compound (II), which is an intermediate of the present invention, by reacting with the condensing agent mentioned in step c. The reaction temperature and reaction time may be the same as in step c.

(Step e) Compound (7) is synthesized by deprotecting the protecting group Rp for the amino group of compound (II) in the same manner as in step b.

(Step f) Compound (7) and carboxylic acid or sulfonic acid represented by formula (8), or their reactive derivative (9) (wherein B is halogen, R'A'O- Or R ″ O—, R′A′O— represents a group capable of becoming an acid anhydride, and R ″ O— represents a group capable of becoming an active ester), thereby subjecting the compound (10 ) Is synthesized. As the method, the same method as in step c can be adopted.

(Step g) The compound (I) of the present invention is synthesized by oxidizing the hydroxyl group of the compound (10). Examples of the suitable oxidation method include hypervalent iodine such as periodinane such as 1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3 (1H) -one. The method of using the reagent in dichloromethane at room temperature is adopted. Other suitable methods include a method using excess dimethyl sulfoxide and water-soluble carbodiimide with dichloroacetic acid as a catalyst in an inert solvent such as toluene at room temperature, a method using oxalyl chloride, dimethyl sulfoxide, and alkaline permanganese. Method using potassium acid aqueous solution, method using pyridine sulfur trioxide complex, and chromium (IV) oxide in methylene chloride
A method using a pyridine complex or the like is adopted. Incidentally, -C
When synthesizing the compound (I) of the present invention having a group represented by OR ₈ , particularly when synthesizing a compound in which R ₈ is a hydroxyl group, the group represented by —COR ₈ is substituted by the substitution in each reaction described above. In consideration of the stability of the group, each reaction is carried out in the state of a benzyl ester group, a tert-butyl ester group, a methyl ester group, etc., which are included as specific examples, and if necessary, by a method known per se at the final stage. It is preferable to convert to various R ₈ . Compound (10) can also be synthesized by the method shown in Scheme 2 below.

[0065]

Embedded image

(Step h) The above amine compound (3)
The compound (12) is synthesized by subjecting the carboxylic acid of (11) (a synthesis method is described later) or its reactive derivative to a condensation reaction. As the method, the same method as in step c of scheme 1 can be adopted. (Step i) Compound (10) is synthesized by reacting compound (12) with amine of compound (6) in the same manner as in step d of Scheme 1. Compound (10) can also be synthesized by the method shown in Scheme 3 below.

[0067]

Embedded image

(Step j) The above ester compound (2) is added to
Amine compound (6)
Compound (13) is synthesized by reacting with. (Step k) The compound (14) is synthesized by deprotecting the protecting group Rp for the amino group of the compound (13) in the same manner as in the step b of Scheme 1. (Step l) Compound (14) is reacted with carboxylic acid (11) or a reactive derivative thereof in the same manner as in step c of Scheme 1 to synthesize compound (10). The compound (I) of the present invention can also be synthesized from the compound (II) by the method shown in the following scheme 4.

[0069]

Embedded image

(Step m) Compound (II) is synthesized by the same oxidation method as in Step g of Scheme 1 to synthesize compound (15). (Step n) Compound (16) is synthesized by deprotecting amino-protecting group Rp of compound (15) in the same manner as in step b of scheme 1. (Step o) Compound (16) of the present invention is subjected to condensation reaction with carboxylic acid or sulfonic acid (8) similar to that in step f of Scheme 1 or a reactive derivative (9) thereof to give compound (I) of the present invention. To synthesize. The points to be noted when synthesizing the compound (I) of the present invention having a group represented by —COR ₈ are as described above.

Compound (II) can be further synthesized by the method shown in Scheme 5.

[0072]

Embedded image

(Step p) The above amine compound (14) and R
Compound (18) is synthesized from p-protected proline (17) using a method similar to step c in Scheme 1. (Step q) The amine group (19) is synthesized by deprotecting the amino group-protecting group Rp of the compound (18) in the same manner as in Scheme 1, step b. (Step r) By subjecting the amine compound (19) and a carboxylic acid represented by the formula (20) (obtained in an intermediate step of the synthesis of the carboxylic acid (4) described later) or a reactive derivative thereof to a condensation reaction , Compound (II) is synthesized. As the method, the same method as in step c of scheme 1 can be adopted.

The carboxylic acid (4) and the carboxylic acid (11) are
It synthesizes as follows. First, the amino-protecting group Rp or R ₁ -A- is replaced with an amino acid by the method described above.

[0075]

Embedded image

It is introduced into the amino group of. Then, the carboxyl group of this compound is condensed with the amino group of proline in which the carboxyl group is protected by a protecting group known per se, and then the protecting group of the carboxyl group is removed by a method known per se to synthesize. As the condensation method, the same method as in step c of scheme 1 is adopted.

Compound (I) of the present invention thus synthesized
Can be collected in any purity by appropriately performing known separation and purification means such as concentration, extraction, chromatography, reprecipitation, and recrystallization.

The pharmaceutically acceptable salt of the compound (I) can also be produced by a known method. further,
Various isomers of the compound (I) can also be produced by a known method.

The compound (I) of the present invention and a pharmacologically acceptable salt thereof have an excellent inhibitory effect on the chymase group on mammals (eg, humans, dogs, cats, etc.).
Therefore, the compound (I) of the present invention and a pharmacologically acceptable salt thereof are useful as inhibitors of a chymase group including human heart chymase, and diseases thought to involve angiotensin II (for example, hypertension and heart). Hypertrophy, myocardial infarction, arteriosclerosis, diabetic and non-diabetic renal disease, PT
It is useful for the prevention / treatment of various diseases caused by chymase including the prevention / treatment of vascular restenosis after CA).

When the compound (I) of the present invention and a pharmacologically acceptable salt thereof are used as a medicine, a pharmacologically acceptable carrier or the like is used, and granules, tablets, capsules, injections, ointments, creams, aerosols are used. The pharmaceutical composition can be administered orally or parenterally in the above manner.
An effective amount of compound (I) or a pharmacologically acceptable salt thereof is blended in the above preparation.

The dose of the compound (I) and a pharmacologically acceptable salt thereof varies depending on the administration route, symptoms of the patient, body weight, age and the like, and can be appropriately set according to the purpose of administration. Usually, when orally administered to an adult, it is preferable to administer 0.01 to 1000 mg / kg body weight / day, preferably 0.05 to 500 mg / kg body weight / day, divided into 1 to several times a day.

[0082]

EXAMPLES The present invention will be described more specifically with reference to Reference Examples and Examples, but the present invention is not limited to these. In addition, ¹ H-NMR was measured at 200 MHz or 500 MHz. The chemical shift of ¹ H-NMR is tetramethylsilane (TMS) as an internal standard.
And the relative delta (δ) value was expressed in parts per million (ppm). The coupling constant indicates trivial multiplicity in Hertz (Hz), s (singlet), d (doublet), t (triplet), q (quartet),
It was expressed as m (multiplet), dd (doublet of doublets), brs (broad singlet), and the like.

Example 1 N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-benzylamine (Step 1) 4 (S) -amino-2,2-difluoro-3 (R) -hydroxy-5- Synthesis of phenylvaleric acid ethyl ester hydrochloride 4 (S)-[N- (tert-butyloxycarbonyl)] amino-2,2-difluoro described in Thairivongs et al. (J. Med. Chem. 1986, 29, 2080). -3 (R) -hydroxy-5-phenylvaleric acid ethyl ester 580 mg
(1.55 mmol) at 0 ° C. in a nitrogen atmosphere at 4N
Hydrochloric acid (dioxane solution 3.9 ml) was added. After stirring at room temperature for 16 hours, the solvent was evaporated under reduced pressure, and the residue was evaporated.
Tellurium was added to collect the precipitated powder, and 470 mg of the target compound was obtained as a white powder. Yield 98% mp. 161.
0-171.0 ° C

(Step 2) 4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,
Synthesis of 2-difluoro-3 (R) -hydroxy-5-phenylvaleric acid ethyl ester N- (tert-butyloxycarbonyl) -L-valyl-L-proline 95 mg (0.30 m under a nitrogen atmosphere.
mol) in a dichloromethane solution (2 ml), triethylamine 42 μl, benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate 133 mg, and the target compound 9 of step 1
0 mg (0.30 mmol) was added. After stirring at room temperature for 16 hours, the reaction product was diluted with 20 ml of dichloromethane, 20 ml of 1N hydrochloric acid aqueous solution × 2, saturated aqueous sodium bicarbonate solution 20.
After washing with water and saturated saline, the mixture was dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was subjected to flash column chromatography (hexane: ethyl acetate = 3:
Purification in 2) yielded 140 mg of the desired compound as a colorless amorphous. Yield 85% ¹ H-NMR (CDCl ₃ ): δ7.24 (6H, m), 5.0-5.5 (2H,
m), 4.0-4.7 (6H, m), 3.3-4.0 (2H, m), 2.8-3.3 (2H,
m), 1.7-2.5 (5H, m), 1.42 (9H, s), 1.29 (3H, t, J =
7.1 Hz), 0.9-1.1 (6 H, m) IR (KBr): 3250, 2900, 1700 cm ^-1

(Step 3) 4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,
Synthesis of 2-difluoro-3 (R) -hydroxy-5-phenylvaleric acid sodium salt 650 mg (1.14 mm) of the target compound of step 2 at room temperature
1N aqueous sodium hydroxide solution (1.14 ml) was added to a tetrahydrofuran solution (2 ml) of ol) and stirred for 2 hours. Tetrahydrofuran was distilled off under reduced pressure, and the remaining aqueous layer was freeze-dried to obtain 660 mg of the target compound as a colorless amorphous substance. Yield: 100% ¹ H-NMR (CDCl ₃ ): δ 7.40 (1 H, d, J = 8.9 Hz), 7.1
-7.35 (5H, m), 6.72 (2H, m), 4.41 (1H, m), 4.11 (1
H, brq, J = 8.2 Hz), 3.99 (1H, t, J = 8.2 Hz), 3.5-3.8
(3H, m), 2.70 (2H, m), 1.7-2.1 (5H, m), 1.35 (9H,
s), 0.87 (6H, m) IR (KBr): 3250, 2900,1630, 1500, 1420 cm ^-1

(Step 4) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3 (R) -hydroxy-5-phenylvaleryl] -N-benzylamine 100 mg of the target compound of step 3 in a nitrogen stream at room temperature
Dichloromethane solution (0.18 mmol) (1 ml)
55 mg of 1-hydroxybenzotriazole (0.
41 mmol), dicyclohexylcarbodiimide 47
mg (0.23 mmol) and benzylamine 28 m
g (0.27 mmol) was added, and the mixture was stirred for 16 hours.
The reaction product was diluted with 10 ml of dichloromethane, washed with 10 ml of a 1N hydrochloric acid aqueous solution, 10 ml of a saturated aqueous sodium hydrogen carbonate solution and 10 ml of a saturated saline solution, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by flash column chromatography (hexane: ethyl acetate = 1: 1) to obtain 60 mg of the target compound as a colorless amorphous substance. Yield 54% ¹ H-NMR (CDCl ₃ ): 7.53 (1H, br), 7.1-7.4 (10H,
m), 6.45 (1H, d, J = 7.9 Hz), 5.14 (1H, d, J = 8.6 Hz),
5.06 (1H, d, J = 8.0 Hz), 4.57 (1H, dd, J = 14.4, 6.2
Hz), 4.38 (1H, dd, J = 14.4, 5.5 Hz), 4.23 (1H, dd, J
= 8.6, 6.3 Hz), 4.18 (1H, dd, J = 8.5, 4.1 Hz), 4.13
(1H, m), 3.92 (1H, brq), 3.72 (1H, m), 3.55 (1H,
m), 3.02 (1H, dd, J = 13.8, 8.4 Hz), 2.90 (1H, dd, J =
13.8, 7.1 Hz), 2.10 (1H, m), 2.03 (1H, m), 1.93 (2
H, m), 1.75-1.90 (1H, m), 1.44 (9H, s), 0.9-1.0 (6
H, m) IR (KBr): 3250, 2900, 1680, 1620 cm ^-1 MS (SIMS) m / z 631 (MH ⁺ )

(Step 5) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-benzylamine 30 mg (0.
05 mmol) in dichloromethane (0.5 ml)
To the above, 40 mg (0.10 mmol) of Dess-Martin reagent was added and stirred for 16 hours. The reaction product was diluted with 10 ml of ether, 10 ml of saturated aqueous sodium hydrogen carbonate containing 2.5 g of sodium thiosulfate was added, and the mixture was further stirred for 2 hours. The separated organic layers were collected, saturated aqueous sodium hydrogen carbonate solution 10 ml, water 10 m.
l, washed with 10 ml of saturated saline and dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure to obtain 28 mg of the title compound as a colorless amorphous substance. Yield 93% ¹ H-NMR (DMSO-d ₆ ): δ 9.65 (1 H, t, J = 6.0 Hz), 8.
48 (1H, d, J = 7.1 Hz), 7.35-7.25 (10H, m), 6.72 (1
H, d, J = 8.5 Hz), 4.98 (1H, ddd, J = 8.6, 7.1. 3.8 H
z), 4.36 (2H, m), 4.34 (1H, m), 3.97 (1H, m), 3.67
(1H, m), 3.53 (1H, m), 3.15 (1H, dd, J = 14.4, 3.9 H
z), 2.81 (1H, dd, J = 14.4, 8.9 Hz), 1.99 (1H, m), 1.
88 (1H, m), 1.84 (2H, m), 1.75 (1H, m), 1.36 (9H,
s), 0.95-0.80 (6H, m) IR (KBr): 3425, 3250, 2950, 1750, 1700, 1635, 14
95, 1430, 1365, 1160cm ^-1 MS (SIMS) m / z 629 (MH ⁺ )

(Example 2) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-benzyl-N-methylamine (Step 1) N- [4 (S)-[N- [N- (tert-butyl Oxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3 (R) -hydroxy-5-phenylvaleryl] -N-benzyl-N-methylamine Using benzylmethylamine instead of benzylamine, shown in step 4 of Example 1. Was synthesized by the method described above. For purification, flash column chromatography (hexane: ethyl acetate = 1: 2) was used to obtain the target compound as a colorless amorphous substance. Yield 37% ¹ H-NMR (CDCl ₃ ): δ7.1-7.4 (10H, m), 5.29 (1H,
d, J = 9.4 Hz), 4.5-4.7 (2H, m), 4.0-4.4 (3H, m), 3.
3-3.8 (3H, m), 3.04 (3H, t, J = 2.1 Hz), 2.9-3.0 (2H,
m), 2.33 (1H, m), 1.7-2.1 (5H, m), 1.42 (9H, s),
0.8-1.1 (6H, m)

(Step 2) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-benzyl-N-methylamine Synthesized from the compound obtained in Step 1 according to the method shown in Step 5 of Example 1, The title compound was obtained as a yellow amorphous. Yield 79% ¹ H-NMR (DMSO-d ₆ ): δ 8.48 (1H), 7.2-7.4 (10H,
m), 6.74 (1H), 5.00 (1H), 4.65 (1H, d, J = 14.8 H
z), 4.46 (1H, d, J = 14.8 Hz), 4.37 (1H, dd, J = 8.2,
4.0 Hz), 3.97 (1H, m), 3.65 (1H, m), 3.53 (1H, m),
3.14 (1H, m), 2.91 (3H, s), 2.89 (1H, dd, J = 14.4,
9.0 Hz), 2.91 (3H, s), 2.89 (1H, dd, J = 14.4, 9.0 H
z), 1.6-2.1 (5H, m), 1.36 (9H, s), 0.8-0.9 (6H, m) IR (KBr): 3250, 2950, 1650, 1160, 690 cm ^-1 MS (CI) m / z 643 (MH ⁺ )

(Example 3) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-phenethylamine (Step 1) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L -Valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3 (R) -hydroxy-5-phenylvaleryl] -N-phenethylamine It was synthesized by the method shown in step 4 of Example 1 using phenethylamine instead of benzylamine. For purification, flash column chromatography (hexane: ethyl acetate = 1: 2) was used to obtain the target compound as a colorless amorphous substance. Yield 41% ¹ H-NMR (CDCl ₃ ): δ 7.1-7.4 (12H, m), 6.2 (1H,
d, J = 7.4 Hz), 5.0-5.2 (2H, m), 4.0-4.3 (3H, m), 3.6
5-3.90 (2H, m), 3.4-3.65 (2H, m), 2.8-3.1 (4H, m),
1.7-2.3 (5H, m), 1.45 (9H, s), 0.8-1.1 (6H, m) IR (KBr): 3300, 2900, 1680, 1620, 1160, 690 cm ^-1

(Step 2) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-phenethylamine The compound obtained in Step 1 was synthesized according to the method shown in Step 5 of Example 1, and the title compound was yellow amorphous. Got as. Yield 95% ¹ H-NMR (DMSO-d ₆ ): δ9.18 (1 H, t, J = 5.6 Hz), 8.
46 (1H, d, J = 7.1 Hz), 7.15-7.35 (10H, m), 6.72 (1
H, d, J = 8.4 Hz), 4.92 (1H, m), 4.35 (1H, dd, J = 8.3,
4.1 Hz), 3.96 (1H, t, J = 8.0 Hz), 3.65 (1H, m), 3.5
2 (1H, m), 3.37 (2H, dd, J = 14.1, 6.3 Hz), 3.13 (1H,
dd, J = 14.5, 4.3 Hz), 2.7-2.82 (3H, m), 1.6-2.1 (5
H, m), 1.36 (9H, s), 0.81-0.87 (6H, m) IR (KBr): 3250, 2950, 1680, 1620, 1160, 690 cm ^-1 MS (CI) m / z 643 (MH ⁺ )

Example 4 N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] glycine methyl ester (Step 1) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L- Valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3 (R) -hydroxy-5-phenylvaleryl] glycine methyl ester It was synthesized by the method shown in Step 4 of Example 1 using glycine methyl ester instead of benzylamine. For purification, flash column chromatography (hexane: ethyl acetate = 1: 3) was used to obtain the target compound as a colorless amorphous substance. Yield 74% ¹ H-NMR (CDCl ₃ ): δ 8.90 (1H, br), 7.92 (1H, d,
J = 6.9 Hz), 7.1-7.3 (5H, m), 5.97 (1H, d, J = 11.0 H
z), 5.05 (1H, d, J = 9.5 Hz), 4.50 (1H, dd, J = 17.9,
8.1 Hz), 4.1-4.3 (3H, m), 3.6-3.8 (3H, m), 3.74 (3
H, s), 3.0-3.3 (2H, m), 2.15-2.4 (1H, m), 1.9-2.15
(4H, m), 1.43 (9H, s), 1.01 (3H, d, J = 6.7 Hz), 0.9
3 (3H, d, J = 6.7 Hz) IR (KBr): 3300, 2950, 1740, 1690, 1620, 1160, 70
0 cm ^-1

(Step 2) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] glycine methyl ester The compound obtained in Step 1 was synthesized according to the method shown in Step 5 of Example 1 to give the title compound as a colorless amorphous. Obtained. Yield 73% ¹ H-NMR (DMSO-d ₆ ): δ9.53 (1 H, t, J = 5.8 Hz), 8.
44 (1H, d, J = 7.1 Hz), 7.15-7.30 (5H, m), 6.72 (1H,
d, J = 8.5 Hz), 4.92 (1H, br), 4.34 (1H, m), 3.8-4.0
(3H, m), 3.55-3.70 (1H, m), 3.63 (3H, s), 3.51 (1
H, m), 3.15 (1H, dd, J = 14.7, 4.2 Hz), 2.80 m (1H, d
d, J = 14.7, 9.1 Hz), 2.00 (1H, m), 1.75-1.95 (3H,
m), 1.65-1.75 (2H, m), 1.35 (9H, s), 0.81-0.86 (6H,
m) IR (KBr): 3300, 2950, 1750, 1690, 1620, 1510, 14
40, 1160, 900 cm ^-1 MS (SIMS) m / z 611 (MH ⁺ )

(Example 5) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] glycine benzyl ester (Step 1) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L- Valyl-L-prolyl] amino]
-2,2-Difluoro-3 (R) -hydroxy-5-phenylvaleryl] glycine benzyl ester was synthesized by the method shown in Step 4 of Example 1 using glycine benzyl ester instead of benzylamine. For purification, flash column chromatography (hexane: ethyl acetate = 1: 1) was used to obtain the target compound as a colorless amorphous substance. Yield 85% ¹ H-NMR (CDCl ₃ ): δ8.91 (1H, br), 7.86 (1H, d,
J = 6.9 Hz), 7.1-7.4 (10H, m), 5.98 (1H, d, J = 11.0 H
z), 5.20 (1H, d, J = 7.5 Hz), 5.10 (1H, d, J = 7.5 Hz),
5.03 (1H, br), 4.54 (1H, dd, J = 9.7, 8.0 Hz), 4.0-
4.3 (3H, m), 3.6-3.9 (4H, m), 3.0-3.3 (2H, m), 2.2
5 (1H, br), 1.94 (4H, m), 1.43 (9H, s), 1.01 (3H,
d, J = 6.7 Hz), 0.93 (3H, d, J = 6.7 Hz) IR (KBr): 3300, 2970, 1700, 1630, 1170, 700 cm ^-1

(Step 2) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] glycine benzyl ester The compound obtained in Step 1 was synthesized according to the method shown in Step 5 of Example 1 to give the title compound as a colorless amorphous substance. Obtained. Yield 93% ¹ H-NMR (CDCl ₃ ): δ8.73 (1H, br), 7.71 (1H, d,
J = 6.7 Hz), 7.15-7.37 (11H, m), 5.18 (2H, s), 5.08
(1H, d, J = 9.3 Hz), 4.31-4.35 (2H, m), 4.23 (1H, dd,
J = 9.1, 6.5 Hz), 3.97 (1H, dd, J = 17.9, 5.0 Hz), 3.6
9 (1H, brq), 3.61 (1H, m), 3.34 (1H, dd, J = 14.4, 4.
8 Hz), 3.19 (1H, dd, J = 14.4, 9.7 Hz), 2.09 (1H, m),
1.78-1.93 (4H, m), 1.43 (9H, s), 0.91 (3H, d, J =
6.7 Hz), 0.87 (3H, d, J = 6.7 Hz) IR (KBr): 3350, 3000, 1750, 1700, 1630, 1170, 70
0 cm ^-1 MS (SIMS) m / z 687 (MH ⁺ )

Example 6 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro- Synthesis of 3-oxo-5-phenylvaleryl] amino] propionic acid benzyl ester (Step 1) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl] -L-prolyl] amino] -2,2-difluoro-3 (R) -hydroxy-
Synthesis of 5-phenylvaleryl] amino] propionic acid benzyl ester [beta] -alanine benzyl ester was used in place of benzylamine, and was synthesized by the method shown in step 4 of Example 1. For purification, flash column chromatography (hexane: ethyl acetate = 1: 2) was used to obtain the target compound as a colorless amorphous substance. Yield 65% ¹ H-NMR (CDCl ₃ ): δ7.82 (1H, br), 7.1-7.5 (10
H, m), 5.37 (1H, d, J = 8.8 Hz), 5.15 (1H, d, J = 8.2 H
z), 5.08 (1H, d, J = 12.6 Hz), 5.00 (1H, d, J = 12.6 H
z), 4.1-4.3 (3H, m), 3.3-4.0 (4H, m), 2.9-3.2 (2H,
m), 2.67 (2H, t, J = 6.2 Hz), 2.12 (2H, m), 1.88 (3
H, m), 1.46 (9H, s), 0.98 (3H, d, J = 6.8 Hz), 0.92
(3H, d, J = 6.8 Hz) IR (KBr): 3300, 3000, 1690, 1630, 1170, 700 cm ^-1

(Step 2) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro-3 Synthesis of -oxo-5-phenylvaleryl] amino] propionic acid benzyl ester The compound obtained in Step 1 was synthesized according to the method shown in Step 5 of Example 1 to obtain the title compound as a colorless amorphous. Yield 81% ¹ H-NMR (DMSO-d ₆ ): δ9.14 (1 H, t, J = 5.4 Hz), 8.4
6 (1H, d, J = 7.0Hz), 7.2-7.4 (10H, m), 6.67 (1H, d,
J = 8.4Hz), 5.08 (2H, s), 4.91 (1H, m), 4.34 (1H, d
d, J = 4.1, 4.2Hz), 3.96 (1H, t, J = 8.1 Hz), 3.66 (1H,
brq), 3.51 (1H, brq), 3.42 (2H, brq), 3.15 (1H, d
d, J = 14.4, 4.4Hz), 2.82 (1H, dd, J = 14.4, 9.0Hz),
2.61 (2H, t, J = 7.1Hz), 1.99 (1H, m), 1.80-1.90 (3H,
m), 1.72 (1H, m), 1.36 (9H, s), 0.81-0.85 (6H, m) IR (KBr): 3300, 3000, 1690, 1630, 1170, 700 cm ^-1

(Example 7) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -L-proline benzyl ester (Step 1) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl)] -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3 (R) -hydroxy-5-phenylvaleryl] -L-proline benzyl ester As shown in step 4 of Example 1, using L-proline benzyl ester instead of benzylamine. It was synthesized by the method. For purification, flash column chromatography (hexane: ethyl acetate = 2: 3) was used to obtain the target compound as a colorless amorphous substance. Yield 51% ¹ H-NMR (CDCl ₃ ): δ7.1-7.4 (10H, m), 5.26 (1H,
d, J = 9.3 Hz), 5.19 (1H, d, J = 12.3 Hz), 5.08 (1H,
d, J = 12.3 Hz), 4.4-4.6 (2H, m), 4.31 (2H, m), 4.0-
4.2 (1H, m), 3.5-4.0 (4H, m), 2.94 (2H, m), 1.7-2.4
(9H, m), 1.42 (9H, s), 1.00 (3H, d, J = 6.8 Hz), 0.9
3 (3H, d, J = 6.8 Hz) IR (KBr): 3300, 2970, 1740, 1650, 1440, 1170, 70
0 cm ^-1

(Step 2) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -L-proline benzyl ester The compound obtained in Step 1 was synthesized according to the method shown in Step 5 of Example 1 to give the title compound. Obtained as a yellow amorphous. Yield 100% ¹ H-NMR (DMSO-d ₆ ): δ 8.39 (1 H, d, J = 7.0 Hz),
7.2-7.4 (10H, m), 6.68 (1H, br), 5.15 (1H, d, J = 12.
6 Hz), 5.11 (1H, d, J = 12.6 Hz), 5.01 (1H, m), 4.49
(1H, m), 4.36 (1H, m), 3.97 (1H, t, J = 8.1 Hz), 3.4-
3.7 (4H, m), 3.13 (1H, dd J = 14.5, 4.5 Hz), 2.84 (1
H, dd, 14.5, 8.8 Hz), 1.6-2.2 (9H, m), 1.36 (9H,
s), 0.86 (3H, d, J = 6.4 Hz), 0.83 (3H, d, J = 6.4 Hz) IR (KBr): 3300, 2970, 1750, 1650, 1440, 1170, 70
0 cm ^-1 MS (SIMS) m / z 727 (MH ⁺ )

(Example 8) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -L-phenylalanine benzyl ester (Step 1) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl)] -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3 (R) -hydroxy-5-phenylvaleryl] -L-phenylalanine benzyl ester As shown in Step 4 of Example 1 using L-phenylalanine benzyl ester instead of benzylamine. It was synthesized by the method. For purification, flash column chromatography (hexane: ethyl acetate = 1: 1) was used to obtain the target compound as a colorless amorphous substance. Yield 5
0% ¹ H-NMR (CDCl ₃ ): δ8.85 (1H, d, J = 7.0 Hz), 7.79
(1H, d, J = 6.8 Hz), 7.1-7.4 (15H, m), 6.05 (1H, d,
J = 11.0 Hz), 5.19 (1H, d, J = 12.2 Hz), 5.09 (1H, d,
J = 12.2 Hz), 4.8-5.1 (2H, m), 4.1-4.3 (3 H, m), 3.5-
3.9 (3H, m), 3.0-3.4 (4H, m), 2.27 (1H, br), 1.96
(4H, m), 1.37 (9H, s), 0.99 (3H, d, J = 6.7 Hz), 0.88
(3H, d, J = 6.7 Hz) IR (KBr): 3300, 2970, 1700, 1630, 1170, 700 cm ^-1

(Step 2) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -L-phenylalanine benzyl ester The title compound was synthesized from the compound obtained in Step 1 according to the method shown in Step 5 of Example 1. Obtained as a colorless amorphous. Yield 93% ¹ H-NMR (DMSO-d ₆ ): δ 9.62 (1 H, d, J = 7.7 Hz), 8.3
9 (1H, d, J = 7.0Hz), 7.1-7.4 (15H, m), 6.68 (1H, d,
J = 8.5Hz), 5.10 (2H, s), 4.93 (1H, m), 4.61 (1H, d
d, J = 6.3, 7.5 Hz), 4.35 (1H, m), 3.96 (1H, t, J = 8.2
Hz), 3.65 (1H, m), 3.50 (1H, m), 3.0-3.2 (3H, m),
2.70 (1H, dd, J = 14.4, 9.3 Hz), 1.70-2.10 (5H, m),
1.36 (9H, s), 0.81-0.85 (6H, m) IR (KBr): 3300, 2970, 1690, 1630, 1170, 700 cm ^-1 MS (SIMS) m / z 777 (MH ⁺ )

(Example 9) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -D-phenylalanine benzyl ester (Step 1) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3 (R) -hydroxy-5-phenylvaleryl] -D-phenylalanine benzyl ester As shown in Step 4 of Example 1, using D-phenylalanine benzyl ester instead of benzylamine. It was synthesized by the method. For purification, flash column chromatography (hexane: ethyl acetate = 1: 1) was used to obtain the target compound as a colorless amorphous substance. Yield 7
7% ¹ H-NMR (CDCl ₃ ): δ8.01 (1H, d, J = 8.5 Hz), 7.0
-7.5 (16H, m), 6.13 (1H, d, J = 7.3 Hz), 5.42 (1H, d,
J = 9.2 Hz), 5.25 (1H, d, J = 12.2 Hz), 5.18 (1H, d, J
= 12.2 Hz), 5.10 (1H, br), 4.93 (1H, m), 4.0-4.4 (2
H, m), 3.71 (1H, br), 3.57 (2H, br), 2.8-3.4 (4H,
m), 1.8-2.4 (5H, m), 1.46 (9H, s), 0.8-1.0 (6H, m) IR (KBr): 3350, 1680, 1620, 1160, 700 cm ^-1

(Step 2) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -D-phenylalanine benzyl ester The compound obtained in Step 1 was synthesized according to the method shown in Step 5 of Example 1 to give the title compound. Obtained as a yellow amorphous. Yield 95% ¹ H-NMR (DMSO-d ₆ ): δ9.58 (1 H, d, J = 7.5 Hz), 8.
43 (1H, d, J = 7.0 Hz), 7.1-7.4 (15H, m), 6.72 (1H,
d, J = 8.4 Hz), 5.10 (1H, d, J = 12.6 Hz), 5.07 (1H, d,
J = 12.6 Hz), 4.86 (1H, br), 4.60 (1H, m), 4.33 (1H,
m), 3.95 (1H, m), 3.64 (1H, m), 3.50 (1H, m), 3.0-
3.2 (3H, m), 2.72 (1H, dd, J = 14.5, 9.3 Hz), 1.7-2.
1 (5H, m), 1.34 (9H, s), 0.80-0.85 (6H, m) IR (KBr): 3250, 2950, 1690, 1620, 1490, 1420, 11
60, 700 cm ^-1 MS (SIMS) m / z 777 (MH ⁺ )

Example 10 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro- Synthesis of 3-oxo-5-phenylvaleryl] amino] -3-phenylpropionic acid benzyl ester (Step 1) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl)] -L-valyl-L-prolyl] amino] -2,2-difluoro-3 (R) -hydroxy-
Synthesis of 5-phenylvaleryl] amino] -3-phenylpropionic acid benzyl ester Synthesized by the method shown in step 4 of Example 1 using 3-amino-3-phenylpropionic acid benzyl ester instead of benzylamine. . For purification, flash column chromatography (hexane: ethyl acetate = 1: 1) was used, and the target compound Rf = 0.50 (hexane: ethyl acetate = 1: 2) (compound A) and Rf =
0.43 (hexane: ethyl acetate = 1: 2) (Compound B) was obtained as a colorless amorphous substance. Yield 3
6% (Compound A), 42% (Compound B) (Compound A) ¹ H-NMR (CDCl ₃ ): δ8.77 (1H, d, J
= 9.2 Hz), 7.61 (1H, d, J = 7.0 Hz), 7.1-7.5 (15H, m),
6.08 (1H, d, J = 10.7 Hz), 5.49 (1H, t, J = 8.6 Hz),
5.09 (1H, d, J = 12.2 Hz), 4.93 (1H, d, J = 12.2 Hz),
4.22 (2H, m), 4.08 (1H, m), 3.6-3.9 (3H, m), 2.9-3.
3 (3H, m), 2.79 (1H, dd, J = 14.2, 2.9 Hz), 2.26 (1
H, m), 1.8-2.2 (4H, m), 1.43 (9H, s), 0.8-1.1 (6H,
m) IR (KBr): 3350, 2950, 1680, 1620, 1160, 690 cm ^-1 (Compound B) ¹ H-NMR (CDCl ₃ ): δ8.42 (1H, d, J
= 8.0 Hz), 7.1-7.5 (15H, m), 6.93 (1H, m), 5.83 (1H,
d, J = 7.3 Hz), 5.4-5.5 (2H, m), 5.09 (2H, s), 4.0-
4.3 (3H, m), 3.4-4.0 (2H, m), 3.1-3.4 (2H, m), 2.85
-3.1 (2H, m), 2.76 (1H, dd, J = 12.6, 7.0 Hz), 1.5-
2.3 (5H, m), 1.31 (9H, s), 0.8-1.1 (6H, m) IR (KBr): 3300, 2900, 1680, 1620, 1160, 690 cm ^-1

(Step 2) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro-3 Synthesis of -oxo-5-phenylvaleryl] amino] -3-phenylpropionic acid benzyl ester From compound A obtained in Step 1, Step 5 in Example 1
The title compound was obtained as a yellow amorphous by synthesizing according to the method shown in. Yield 86% ¹ H-NMR (DMSO-d ₆ ): δ9.63 (1 H, d, J = 8.3 Hz),
8.45 (1H, d, J = 9.6 Hz), 7.20-7.34 (15H, m), 6.73
(1H, d, J = 8.3 Hz), 5.30 (1H, dd, J = 15.3, 8.1 Hz),
5.02 (2H, s), 4.90 (1H, m), 4.33 (1H, m), 3.95 (1
H, m), 3.65 (1H, m), 3.49 (1H, m), 3.0-3.15 (2H,
m), 2.94 (1H, m), 2.76 (1H, dd, J = 14.5, 9.3 Hz), 1.
6-2.0 (5H, m), 1.34 (9H, s), 0.8-0.9 (6H, m) IR (KBr): 3300, 2950, 1680, 1620, 1160, 690 cm ^-1 MS (SIMS) m / z 777 (MH ⁺ )

Example 11 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro- Synthesis of 3-oxo-5-phenylvaleryl] amino] -3-phenylpropionic acid benzyl ester (epimer of 3-phenyl group of the title compound of Example 10) Compound B obtained in Step 1 of Example 10 The compound was synthesized according to the method shown in Step 5 of Example 1 to give the title compound as a yellow amorphous substance. Yield 88% ¹ H-NMR (DMSO-d ₆ ): δ 9.69 (1 H, d, J = 8.3 Hz), 8.
45 (1H, d, J = 7.0 Hz), 7.1-7.4 (15H, m), 6.70 (1H,
d, J = 8.7 Hz), 5.29 (1H, dd, J = 15.0, 8.3 Hz), 5.06
(1H, d, J = 3.5 Hz), 5.04 (1H, d, J = 3.5 Hz), 4.95 (1
H, m), 4.34 (1H, dd, J = 8.4, 4.0 Hz), 3.94 (1H, t, J
= 8.2 Hz), 3.64 (1H, m), 3.47 (1H, m), 3.07 (1H, dd,
J = 14.5, 4.2 Hz), 3.04 (1H, dd, J = 16.0, 8.7 Hz), 2.
94 (1H, dd, J = 16.0, 6.5 Hz), 2.73 (1H, dd, J = 14.7,
9.0 Hz), 1.5-2.0 (5H, m), 1.35 (9H, s), 0.8-0.9 (6
H, m) IR (KBr): 3250, 2950, 1690, 1610, 1160, 690 cm ^-1 MS (SIMS) m / z 777 (MH ⁺ )

(Example 12) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] glycine A solution of the title compound of Example 5 in tetrahydrofuran (1 m
To l), 10 mg of palladium black was added, and the mixture was stirred under a hydrogen atmosphere for 5 hours at room temperature. The reaction solution was filtered through Celite, and the filtrate was concentrated under reduced pressure. Ether-hexane was added to the residue, and the precipitated powder was collected to obtain 39 mg of the title compound as a white powder. Yield 90% ¹ H-NMR (CDCl ₃ ): δ8.41 (1H, br), 7.65 (1H, d,
J = 6.4 Hz), 7.1-7.4 (6H, m), 5.15 (1H, d, J = 9.0 H
z), 3.6-4.6 (6H, m), 3.32 (1H, dd, J = 14.2, 5.1 Hz),
3.17 (1H, dd, J = 14.2, 9.0 Hz), 1.6-2.1 (5H, m), 1.
42 (9H, S), 0.88 (6H, m) IR (KBr): 3300, 3000, 1690, 1630, 1160, 700cm ^-1 MS (SIMS) m / z 597 (MH ⁺ )

Example 13 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro- Synthesis of 3-oxo-5-phenylvaleryl] amino] propionic acid From the compound obtained in Example 6, the title compound was obtained as a white powder by the method shown in Example 12. Yield 80% ¹ H-NMR (DMSO-d ₆ ): δ 12.0-12.8 (1H, br), 9.10
(1H, brs), 8.46 (1H, d, J = 6.9 Hz), 7.1-7.3 (5H, m),
6.72 (1H, d, J = 8.0 Hz), 4.92 (1H, br), 4.34 (1H, b
r), 3.96 (1H, t, J = 8.1 Hz), 3.65 (1H, m), 3.52 (1H,
m), 3.2-3.5 (2H, m), 3.14 (1H, dd, J = 14.3, 4.1 H
z), 2.80 (1H, dd, J = 14.3, 9.0 Hz), 2.45 (2H, t, J =
7.3 Hz), 1.95-2.05 (1H, m), 1.8-1.95 (3H, m), 1.72
(1H, m), 1.35 (9H, s), 0.81-0.86 (6H, m) IR (KBr): 3300, 2980, 1710, 1690, 1530, 1170, 70
0 cm ^-1 MS (SIMS) m / z 611 (MH ⁺ )

(Example 14) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -L-proline The title compound was obtained as a white powder from the compound obtained in Example 7 by the method shown in Example 12. . Yield 83% ¹ H-NMR (DMSO-d ₆ ): δ 12.0-13.5 (1H, br), 8.56
(1H, br), 7.1-7.4 (5H, m), 6.73 (1H, d, J = 7.9 Hz),
4.93 (1H, m), 4.33 (2H, m), 3.94 (1H, m), 3.0-3.8
(5H, m), 2.7-3.0 (1H, m), 1.5-2.3 (9H, m), 1.36 (9
H, s), 0.85 (6H, m) IR (KBr): 3300, 2970, 1710, 1660, 1450, 1170, 7
00 cm ^-1 MS (SIMS) m / z 637 (MH ⁺ )

Example 15 N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -L-phenylalanine The title compound was obtained as a white powder from the compound obtained in Example 8 by the method shown in Example 12. . Yield 85% ¹ H-NMR (DMSO-d ₆ ): δ 11.5-13.5 (1H, br), 9.40
(1H, d, J = 8.0 Hz), 8.49 (1H, d, J = 6.0 Hz), 7.23 (1
0H, br), 6.71 (1H, d, J = 8.1 Hz), 4.84 (1H, br), 4.4
2 (1H, m), 4.36 (1H, m), 3.95 (1H, t, J = 8.1 Hz), 3.
61 (1H, m), 3.52 (1H, m), 3.0-3.5 (3H, m), 2.67 (1
H, dd, J = 14.3, 9.4 Hz), 1.6-2.1 (5H, m), 1.34 (9H,
s), 0.84 (6H, br) IR (KBr): 3300, 2970, 1690, 1630, 1160, 700 cm ^-1

Example 16 N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -D-phenylalanine The title compound was obtained as a white powder from the compound obtained in Example 9 by the method shown in Example 12. . Yield 88% ¹ H-NMR (DMSO-d ₆ ): δ 12.0-14.0 (1H, br), 9.35
(1H, d, J = 7.9 Hz), 8.43 (1H, d, J = 7.0 Hz), 7.1-7.4
(10H, m), 6.73 (1H, d, J = 8.3 Hz), 4.88 (1H, m), 4.
46 (1H, m), 4.35 (1H, m), 3.96 (1H, t, J = 8.0 Hz),
3.0-3.8 (5H, m), 2.72 (1H, dd, J = 14.5, 9.2 Hz), 1.6
-2.1 (5H, m), 1.35 (9H, s), 0.8-1.0 (6H, m) IR (KBr): 3250, 2190, 1690, 1620, 1510, 1160cm
^-1

Example 17 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro- Synthesis of 3-oxo-5-phenylvaleryl] amino] -3-phenylpropionic acid The title compound was obtained as a white powder from the compound obtained in Example 10 by the method shown in Example 12. Yield 74% ¹ H-NMR (DMSO-d ₆ ): δ 12.0-12.5 (1H, br), 9.61
(1H, d, J = 7.7 Hz), 8.46 (1H, d, J = 6.7 Hz), 7.1-7.4
(10H, m), 6.72 (1H, d, J = 8.4 Hz), 5.23 (1H, q, J =
7.3 Hz), 4.89 (1H, m), 4.33 (1H, q, J = 3.9 Hz), 3.95
(1H, t, J = 8.0 Hz), 3.63 (1H, m), 3.50 (1H, m), 3.1
0 (1H, dd, J = 14.5, 3.9 Hz), 2.87 (1H, dd, J = 14.4,
8.2 Hz), 2.73-2.78 (2H, m), 1.6-2.1 (5H, m), 1.35
(9H, s), 0.8-0.9 (6H, m) IR (KBr): 3250, 2950, 1680, 1500, 1160, 690 cm ^-1 MS (SIMS) m / z 687 (MH ⁺ )

Example 18 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro- 3-oxo-5-phenylvaleryl] amino] -3-phenylpropionic acid (epimer of 3-phenyl group of the title compound of Example 17) The method shown in Example 12 from the compound obtained in Example 11 The title compound was obtained as a white powder. Yield 76% ¹ H-NMR (DMSO-d ₆ ): δ 12.0-12.7 (1H, br), 9.64
(1H, d, J = 8.0 Hz), 8.43 (1H, d, J = 7.0 Hz), 7.1-7.4
(10H, m), 6.70 (1H, d, J = 8.6 Hz), 5.22 (1H, m), 4.
94 (1H, m), 4.34 (1H, q, J = 4.2 Hz), 3.95 (1H, t, J =
8.2 Hz), 3.65 (1H, dd, J = 8.7, 7.6 Hz), 3.50 (1H,
m), 3.07 (1H, dd, J = 14.6, 4.1 Hz), 2.88 (1H, dd, J =
14.6, 8.4 Hz), 2.7-2.85 (2H, m), 1.50-2.05 (5H, m),
1.35 (9H, s), 0.80-0.90 (6H, m) IR (KBr): 3250, 2950, 1680, 1620, 1510, 1160, 69
0 cm ^-1 MS (SIMS) m / z 687 (MH ⁺ )

Example 19 N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-benzylamine (Step 1) 4 (R) -amino-2,2-difluoro-3 (S) -hydroxy-5- Synthesis of phenylvaleric acid ethyl ester hydrochloride According to the method of Thairivongs et al. N
-(Tert-Butyloxycarbonyl)] amino-
2.0 g (5.4 mmol) of 2,2-difluoro-3 (S) -hydroxy-5-phenylvaleric acid ethyl ester
To the above, 4N hydrochloric acid (dioxane solution 13.5 ml) was added at 0 ° C. under a nitrogen atmosphere. After stirring at room temperature for 16 hours, the solvent was distilled off under reduced pressure, ether was added to the residue, and the precipitated powder was collected to give 1.64 of the target compound as a white powder.
g was obtained. Yield 99% ¹ H-NMR (CDCl ₃ ): δ8.12 (3H, br), 7.2-7.5 (5H,
m), 4.22 (2H, q, J = 7.1 Hz), 3.94 (1H, m), 3.60 (1
H, br), 3.12 (1H, dd, J = 13.5, 4,5 Hz), 2.89 (1H, d
d, J = 13.4, 10.2 Hz), 1.17 (3H, t, J = 7.1 Hz) IR (KBr): 3150, 2850, 1760, 1495, 1100, 1060, 75
5, 700 cm ^-1 [α] _D ²⁵ = ＋36.5 ° (CHCl ₃ , c = 0.97)

(Step 2) 4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,
Synthesis of 2-difluoro-3 (S) -hydroxy-5-phenylvaleric acid ethyl ester According to the method of Step 2 of Example 1, N- (tert-butyloxycarbonyl) -L-valyl-L-proline,
Using the target compound of Step 1, the target compound was obtained as a colorless amorphous. Yield 81% ¹ H-NMR (CDCl ₃ ): δ7.2-7.4 (5H, m), 7.02 (1H,
d, J = 8.2 Hz), 5.21 (1H, d, J = 9.2 Hz), 4.4-4.6 (2H,
m), 4.30 (2H, q, J = 7.1 Hz), 4.2-4.4 (1H, m), 3.9-4.
2 (1H, m), 3.4-3.9 (2H, m), 2.9-3.2 (2H, m), 1.8-2.
3 (5H, m), 1.43 (9H, s), 1.31 (3H, t, J = 7.1 Hz),
0.8-1.1 (6H, m) IR (KBr): 3300, 2950, 1760, 1640,
1500, 1160, 1090 cm ^-1

(Step 3) 4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,
Synthesis of 2-difluoro-3 (S) -hydroxy-5-phenylvaleric acid sodium salt According to the method of Step 3 of Example 1, the target compound of Step 2 was used to obtain the target compound as colorless amorphous. Yield: 100% ¹ H-NMR (DMSO-d ₆ ): δ 7.1-7.4 (5H, m), 6.6-6.8
(2H, br), 4.2-4.35 (1H, m), 4.07 (2H, m), 3.5-3.8
(3H, m), 2.72 (2H, d), 1.6-2.3 (5H, m), 1.38 (9H,
s), 0.94 (3H, d, J = 6.6 Hz), 0.87 (3H, d, J = 6.6 Hz) IR (KBr): 3300, 2950, 1630, 1160 cm ^-1

(Step 4) N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3 (S) -hydroxy-5-phenylvaleryl] -N-benzylamine According to the method of Step 4 of Example 1, the target compound of Step 3 was used and the target compound was colorless amorphous. Got as. Yield 32% ¹ H-NMR (CDCl ₃ ): 7.49 (1H, brt), 7.16-7.46 (10
H, m), 6.72 (1H, d, J = 8.7 Hz), 5.10 (1H, d, J = 9.4 H
z), 4.59 (1H, dd, J = 14.5, 6.2 Hz), 4.36 (1H, dd, J =
14.5, 5.4Hz), 4.21-4.32 (4H, m), 4.10 (1H, m), 4.13
(1H, m), 3.71 (1H, m), 3.52 (1H, m), 2.96 (2H, d,
J = 8.0 Hz), 1.8-2.0 (5H, m), 1.42 (9H, s), 0.84-0.96
(6H, m) IR (KBr): 3300, 2950, 1680, 1625, 1160, 695 cm
^-1

(Step 5) N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-benzylamine 100 mg of the target compound of step 4 in a nitrogen stream at room temperature
Dichloromethane solution (0.16 mmol) (1 ml)
268mg (0.63mmol) of Dess-Martin reagent
Was added and stirred for 16 hours. The reaction product is 10 ml of ether
The mixture was diluted with, and 3 ml of saturated aqueous sodium hydrogen carbonate containing 0.75 g of sodium thiosulfate was added, and the mixture was further stirred for 2 hours. The separated organic layers were collected, saturated aqueous sodium hydrogen carbonate solution 10 ml, water 10 m.
1, washed with 10 ml of saturated saline and dried over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, the obtained residue was dissolved again in 0.8 ml of chloroform, and molecular sieves were added to it at room temperature. After stirring for 2 hours, the mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain 73 mg of the title compound as a colorless amorphous substance. Yield 70% ¹ H-NMR (DMSO-d ₆ ): δ 9.64 (1 H, t, J = 6.0 Hz), 8.
43 (1H, d, J = 8.4 Hz), 7.1-7.4 (10H, m), 6.68 (1H,
d, J = 8.4 Hz), 5.06 (1H, m), 4.41 (1H, dd, J = 15.0,
6.2 Hz), 4.30 (1H, dd, J = 15.0, 6.0 Hz), 4.17 (1H, d
d, J = 4.5, 3.7 Hz), 3.95 (1H, t, J = 8.0 Hz), 3.61 (1
H, dd, J = 16.4, 7.1 Hz), 3.44 (1H, m), 3.17 (1H, dd,
J = 14.0, 3.8 Hz), 2.68 (1H, dd, J = 14.0, 10.5 Hz),
1.6-1.9 (5H, m), 1.33 (9H, s), 0.78-0.89 (6H, m) IR (KBr): 3300, 2950, 1680, 1620, 1440, 1160, 70
0 cm ^-1 MS (SIMS) m / z 629 (MH ⁺ )

Example 20 N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-benzyl-N-methylamine (Step 1) N- [4 (R)-[N- [N- (tert-butyl Oxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3 (S) -hydroxy-5-phenylvaleryl] -N-benzyl-N-methylamine Using benzylmethylamine in place of benzylamine, shown in step 4 of Example 19. Was synthesized by the method described above. For purification, flash column chromatography (hexane: ethyl acetate = 1: 1) was used to obtain the target compound as a colorless amorphous substance. Yield 30% ¹ H-NMR (DMSO-d ₆ ): δ8.43 (1 H, d, J = 8.6 Hz), 7.
18-7.39 (10H, m), 6.69 (1H, d, J = 8.6 Hz), 5.01 (1
H, m), 4.5-4.6 (2H, m), 4.27 (1H, m), 3.93 (1H, t,
J = 8.1 Hz), 3.60 (1H, dd, J = 16.1, 7.2 Hz), 3.42 (1H,
m), 3.17 (1H, dd, J = 10.2, 3.5 Hz), 2.93 (3H, s),
2.78 (1H, m), 1.6-2.0 (5H, m), 1.35 (9H, s), 0.78-
0.86 (6H, m) IR (KBr): 3300, 2950, 1680, 1650, 1160, 695 cm
^-1

(Step 2) N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-benzyl-N-methylamine Synthesized according to the method shown in Step 5 of Example 19 to obtain the title compound as a yellow amorphous substance. . Yield 8
9% ¹ H-NMR (CDCl ₃ ): δ7.1-7.4 (10H, m), 7.05 (1H,
d, J = 8.6 Hz), 5.28 (1H, d, J = 8.6 Hz), 4.5-4.7 (3H,
m), 4.15-4.4 (3H, m), 4.05 (1H, m), 3.3-3.8 (2H,
m), 3.03 (3H, s), 2.80-3.10 (2H, m), 1.8-2.3 (5H,
m), 1.43 (9H, s), 0.86-1.0 (6H, m) IR (KBr): 3300, 2900, 1640, 1160, 695 cm ^-1 MS (SIMS) m / z 643 (MH ⁺ )

Example 21 N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-phenethylamine (Step 1) N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L -Valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3 (S) -hydroxy-5-phenylvaleryl] -N-phenethylamine It was synthesized by the method shown in Step 4 of Example 19 using phenethylamine instead of benzylamine.
For purification, flash column chromatography (hexane: ethyl acetate = 1: 1) was used to obtain the target compound as a colorless amorphous substance. Yield 27% ¹ H-NMR (CDCl ₃ ): δ7.1-7.4 (11H, m), 6.70 (1H,
d, J = 8.6 Hz), 5.14 (1H, d, J = 8.9 Hz), 3.95-4.4 (5
H, m), 3.70 (1H, m), 3.45-3.65 (3H, m), 2.96 (2H,
d, J = 7.7 Hz), 2.85 (2H, t, J = 7.2 Hz), 1.8-2.1 (5H,
m), 1.43 (9H, s), 0.97 (3H, d, J = 6.7 Hz), 0.89 (3H,
d, J = 6.7 Hz) IR (KBr): 3300, 2950, 1680, 1620, 1160, 695 cm ^-1

(Step 2) N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-phenethylamine By the method shown in Step 5 of Example 19, the title compound was obtained as a colorless amorphous. Yield 5
8% ¹ H-NMR (DMSO-d ₆ ): δ9.16 (1 H, t, J = 5.6 Hz), 8.
41 (1H, d, J = 8.5 Hz), 7.15-7.32 (10H, m), 6.72 (1
H, d, J = 8.4 Hz), 5.06 (1H, m), 4.19 (1H, dd, J = 7.9,
4.7 Hz), 3.96 (1H, t, J = 8.0 Hz), 3.63 (1H, dd, J = 1
6.7, 7.4 Hz), 3.43 (2H, m), 3.33 (1H, m), 3.16 (1H,
dd, J = 14.0, 3.8 Hz), 2.78 (2H, t, J = 7.6 Hz), 2.65
(1H, dd, J = 14.0, 10.6 Hz), 1.61-1.92 (5H, m), 1.33
(9 H, s), 0.86 (3H, d, J = 6.6 Hz), 0.81 (3H, d, J =
6.6 Hz) IR (KBr): 3300, 2950, 1670, 1620, 1160, 695 cm ^-1 MS (SIMS) m / z 643 (MH ⁺ )

Example 22 N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] glycine benzyl ester (Step 1) N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L- Valyl-L-prolyl] amino]
-2,2-Difluoro-3 (S) -hydroxy-5-phenylvaleryl] glycine benzyl ester was synthesized by the method shown in Step 4 of Example 19 using glycine benzyl ester instead of benzylamine. For purification, flash column chromatography (hexane: ethyl acetate = 1: 2) was used to obtain the target compound as a colorless amorphous substance. Yield 47% ¹ H-NMR (CDCl ₃ ): δ7.97 (1H, br), 7.13-7.5 (10
H, m), 6.83 (1H, d, J = 8.6 Hz), 5.22 (1H, d, J = 12.3
Hz), 5.19 (1H, d, J = 12.3 Hz), 5.15 (1H, d, J = 9.3 H
z), 4.49 (1H, q, J = 17.9 Hz), 4.29 (1H, dd, J = 3.7,
4.1 Hz), 4.25 (1 H, m), 4.0-4.2 (3H, m), 3.69 (1H,
m), 3.53 (1H, m), 2.97 (2H, d, J = 7.8 Hz), 1.8-2.0
(5H, m), 1.43 (9 H, s), 0.87-0.99 (6H, m) IR (KBr): 3300, 2950, 1750, 1690, 1630, 1160, 70
0 cm ^-1

(Step 2) N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] glycine benzyl ester Step 5 in Example 19 from the compound obtained in Step 1
The title compound was obtained as a yellow amorphous by synthesizing according to the method shown in. Yield 85% ¹ H-NMR (DMSO-d ₆ ): δ9.54 (1 H, t, J = 5.9 Hz), 8.
41 (1H, d, J = 8.5 Hz), 7.20-7.40 (10H, m), 6.74 (1
H, d, J = 8.4 Hz), 5.16 (2H, s), 5.07 (1H, m), 4.21
(1H, dd, J = 8.1, 4.5 Hz), 4.06 (1H, dd, J = 17.4, 5.8
Hz), 3.97 (2H, dd, J = 16.9, 6.0 Hz), 3.59-3.65 (1H,
m), 3.42-3.47 (1H, m), 3.19 (1H, dd, J = 14.0, 3.8 H
z), 2.68 (1H, dd, J = 14.0, 10.6 Hz), 1.6-1.95 (5H,
m), 1.35 (9H, s), 0.81-0.87 (6H, m) IR (KBr): 3300, 2950, 1750, 1690, 1630, 1160, 70
0 cm ^-1 MS (SIMS) m / z 687 (MH ⁺ )

Example 23 3- [N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro- Synthesis of 3-oxo-5-phenylvaleryl] amino] propionic acid benzyl ester (Step 1) 3- [N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl] -L-prolyl] amino] -2,2-difluoro-3 (S) -hydroxy-
Synthesis of 5-phenylvaleryl] amino] propionic acid benzyl ester It was synthesized by the method shown in Step 4 of Example 19 using β-alanine benzyl ester instead of benzylamine. For purification, flash column chromatography (hexane: ethyl acetate = 1: 2) was used to obtain the target compound as a colorless amorphous substance. Yield 47% ¹ H-NMR (CDCl ₃ ): δ7.47 (1H, brt), 7.20-7.40
(10H, m), 6.77 (1H, d, J = 8.6 Hz), 5.28 (1H, d, J = 9.
4 Hz), 5.13 (2H, S), 4.38 (1H, dd, J = 7.8, 4.4 Hz),
4.22-4.31 (2H, m), 4.03-4.08 (2H, m), 3.72 (1H, m),
3.50-3.61 (3H, m), 2.96 (2H, d, J = 7.8 Hz), 2.65 (1
H, d, J = 6.3 Hz), 2.63 (1H, d, J = 6.3 Hz), 1.86-2.08
(5H, m), 1.43 (9H, s), 0.89-1.0 (6H, m) IR (KBr): 3300, 2970, 1690, 1630, 1170, 700 cm ^-1

(Step 2) 3- [N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro-3 Synthesis of -oxo-5-phenylvaleryl] amino] propionic acid benzyl ester The compound was synthesized according to the method shown in Step 5 of Example 19 to give the title compound as a yellow amorphous substance. Yield 9
1% ¹ H-NMR (DMSO-d ₆ ): δ9.15 (1H, t, J = 5.5Hz), 8.4
3 (1H, d, J = 8.4Hz), 7.13-7.37 (10H, m), 6.73 (1H,
d, J = 8.4Hz), 5.09 (2H, s), 5.06 (1H, m), 4.19 (1H,
dd, J = 4.5, 8.1Hz), 3.96 (1H, t, J = 8.0 Hz), 3.62 (1
H, m), 3.3-3.5 (3H, m), 3.18 (1H, dd, J = 14.1, 3.9H
z), 2.67 (1H, dd, J = 13.9, 10.6 Hz), 2.63 (2H, t, J =
7.3Hz), 1.6-1.95 (5H, m), 1.34 (9H, s), 0.8-0.9 (6
H, m) IR (KBr): 3300, 2950, 1690, 1630, 1170, 700 cm ^-1 MS (SIMS) m / z 701 (MH ⁺ )

Example 24 N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] glycine The compound obtained in Example 22 was used for synthesis according to the method shown in Example 12 to obtain the title compound as a white powder. . Yield 60% ¹ H-NMR (DMSO-d ₆ ): δ 12.84 (1H, br), 9.43 (1H,
t, J = 5.7 Hz), 8.56 (1H, br), 7.18-7.27 (5H, m), 6.7
6 (1H, d, J = 8.4 Hz), 5.03 (1H, m), 4.22 (1H, dd, J =
7.7, 4.3 Hz), 3.95 (1H, t, J = 8.1 Hz), 3.82 (2H, b
r), 3.61 (1H, dd, J = 16.4, 7.5 Hz), 3.44 (1H, m), 3.1
9 (1H, dd, J = 14.0, 3.8 Hz), 2.67 (1H, dd, J = 13.9,
10.8 Hz), 1.6-1.95 (5H, m), 1.35 (9H, s), 0.80-0.87
(6H, m) IR (KBr): 3300, 2950, 1750, 1680, 1160 cm ^-1 MS (SIMS) m / z 597 (MH ⁺ )

Example 25 3- [N- [4 (R)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro- Synthesis of 3-oxo-5-phenylvaleryl] amino] propionic acid The compound obtained in Example 23 was synthesized according to the method shown in Example 12 to obtain the title compound as a white powder. Yield 60% ¹ H-NMR (DMSO-d ₆ ): δ 12.25 (1H, br), 9.10 (1H,
t, J = 5.0 Hz), 8.42 (1H, d, J = 8.5 Hz), 7.18-7.30 (5
H, m), 6.74 (1H, d, J = 8.2 Hz), 5.05 (1H, m), 4.19
(1H, dd, J = 7.8, 4.6 Hz), 3.96 (1H, t, J = 7.9 Hz), 3.
62 (1H, dd, J = 16.3, 7.1 Hz), 3.35-3.46 (3H, m), 3.1
8 (1H, dd, J = 14.1, 3.8 Hz), 2.67 (1H, dd, J = 13.8,
10.7 Hz), 2.47 (2H, t, J = 7.5 Hz), 1.6-2.0 (5H, m),
1.35 (9H, s), 0.81-0.88 (6H, m) IR (KBr): 3300, 2950, 1690, 1630, 1160cm ^-1 MS (SIMS) m / z 611 (MH ⁺ )

The compounds obtained in the above Examples 1 to 25 are summarized in Tables 1 and 2. However, for the compounds of Examples 4 to 9, 12 to 16 and 22 to 25 in Table 1 and Table 2, R ₅ and X are represented as —N (R ₅ ) —X.
Also, Gly, β-Ala, Phe, Pro, Bzl are
Glycine, β-alanine, phenylalanine,
Indicates proline and benzyl.

[0130]

[Table 1]

[0131]

[Table 2]

Example 26 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro- Synthesis of 3-oxo-5-phenylvaleryl] amino] benzoic acid benzyl ester (Step 1) 3- [N- [4 (S)-[N- (tert-butyloxycarbonyl) amino] -2,2- Difluoro-3
(R) -Hydroxy-5-phenylvaleryl] amino]
Synthesis of benzoic acid benzyl ester 4 (S) -described in step 1 of Example 1 at room temperature in a nitrogen stream.
It was synthesized from [N- (tert-butyloxycarbonyl)] amino-2,2-difluoro-3 (R) -hydroxy-5-phenylvaleric acid ethyl ester according to the method shown in Step 3 of Example 1 above. (S)-[N- (te
rt-Butyloxycarbonyl) amino] -2,2-difluoro-3 (R) -hydroxy-5-phenylvaleric acid sodium salt 3.39 g (9.24 mmol) in dichloromethane solution (20 ml), 1-hydroxybenzotriazole. 2.87 g (21.2 mmol), 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride 2.66 g (13.9 mmol), and 3
-Aminobenzoic acid benzyl ester 3.15 g (13.
9 mmol) was added and the mixture was stirred for 96 hours. The reaction product was diluted with 15 ml of ethyl acetate and 50 ml of dichloromethane, washed with 100 ml of water, 100 ml of 1N hydrochloric acid aqueous solution, 100 ml of saturated sodium bicarbonate aqueous solution and 100 ml of saturated saline solution, and dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. did. The residue was purified by flash column chromatography (chloroform: methanol = 100: 1) to obtain 3.2 g of the target compound as a colorless amorphous substance. Yield 62% ¹ H-NMR (CDCl ₃ ): 10.66 (1H, br), 8.40 (1H, S),
8.00 (1H, d, J = 8.2 Hz), 7.78 (1H, d, J = 7.8 Hz), 7.
19-7.55 (11H, m), 6.36 (1H, d, J = 9.3 Hz), 6.17 (1H,
br), 5.37 (2H, s), 4.02-4.10 (2H, m), 2.72-2.88 (2
H, m), 1.25 (9H, S) IR (KBr): 3350, 1680, 1280, 1090, 750, 700 cm ^-1

(Step 2) 3- [N- [4 (S) -amino-2,2-difluoro-
Synthesis of 3 (R) -hydroxy-5-phenylvaleryl] amino] benzoic acid benzyl ester hydrochloride 2.13 g (3.84 mmo) of the compound obtained in step 1
To l) was added 4N hydrochloric acid (dioxane solution 9.6 ml) at 0 ° C. under a nitrogen atmosphere. After stirring at room temperature for 1 hour, the solvent was evaporated under reduced pressure, ether and hexane were added to the residue, and the precipitated powder was collected to obtain 2.03 g of the desired compound as a white powder. Yield 100% ¹ H-NMR (DMSO-d ₆ ): 11.02 (1H, s), 8.36 (1H, S),
8.25 (3H, br), 7.99 (1H, d, J = 8.1 Hz), 7.81 (1H,
d, J = 7.8 Hz), 7.27-7.57 (11H, m), 5.38 (2H, s), 4.1
2 (1H, m), 3.67 (1H, br), 3.19 (1H, dd, J = 13.6, 4.5
Hz), 2.95 (1H, dd, J = 13.6, 10.4 Hz) IR (KBr): 3200, 1700, 1280, 1100, 750, 700 cm ^-1

(Step 3) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro-3 (R) -hydroxy-
Synthesis of 5-phenylvaleryl] amino] benzoic acid benzyl ester N- (tert-butyloxycarbonyl) -L-valyl-L-proline 1.01 g (3.23) under nitrogen atmosphere.
mmol) in a dichloromethane solution (10 ml), triethylamine (430 μl), benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate 1.43 g (3.23 mmo)
l) was added, followed by 1.44 g of the compound obtained in step 2.
(2.93 mmol) and 430 μl of triethylamine were added, and the mixture was stirred at room temperature for 64 hours. The reaction product was diluted with 50 ml of dichloromethane, 50 ml of water, 50 ml of 1N hydrochloric acid aqueous solution, 50 ml of saturated aqueous sodium bicarbonate solution, and 50% saturated saline solution.
After washing with ml, the extract was dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by flash column chromatography (hexane: ethyl acetate = 1: 1) to obtain 1.70 g of the objective compound as colorless amorphous. Yield 77% ¹ H-NMR (CDCl ₃ ): 9.21 (1H, s), 8.30 (1H, t, J =
1.7 Hz), 7.98 (1H, d, J = 8.0 Hz), 7.87 (1H, d, J = 8.0
Hz), 7.15-7.48 (11H, m), 6.95 (1H, d, J = 8.0 Hz),
5.36 (2H, s), 5.21 (1H, d, J = 6.0 Hz), 5.07 (1H, br
d, J = 7.9 Hz), 4.34 (1H, m), 4.08-4.29 (2H, m), 3.74
(1H, m), 3.58 (1H, m), 2.93-3.10 (2H, m), 1.70-2.2
0 (5H, m), 1.43 (9H, s), 0.91-1.02 (6H, m) IR (KBr): 3300, 2950, 1700, 1280, 1060, 750, 700
cm ^-1

(Step 4) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro-3 Synthesis of -oxo-5-phenylvaleryl] amino] benzoic acid benzyl ester The compound obtained in Step 3 was synthesized according to the method shown in Step 5 of Example 1. For purification, flash column chromatography (hexane: ethyl acetate = 4:
Using 5) the title compound was obtained as a colorless amorphous. Yield 95% ¹ H-NMR (DMSO-d ₆ ): 11.05 (1H, s), 8.57 (1H, d,
J = 7.8 Hz), 8.39 (1H, br), 7.89 (1H, d, J = 8.2 Hz),
7.79 (1H, d, J = 6.6 Hz), 7.10-7.51 (11H, m), 6.74 (1
H, d, J = 8.7 Hz), 5.36 (2H, s), 5.09 (1H, m), 4.32
(1H, m), 3.95 (1H, m), 3.61 (1H, m), 3.44 (1H, m),
3.13-3.22 (1H, m), 2.72 (1H, dd, J = 10.4, 14.0 Hz),
1.60-2.00 (5H, m), 1.34 (9H, s), 0.73-0.84 (6H, m) IR (KBr): 3300, 2950, 1700, 1620, 1280, 750, 700
cm ^-1

Example 27 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino] -2,2-difluoro- Synthesis of 3-oxo-5-phenylvaleryl] amino] benzoic acid The compound obtained in Example 26 was synthesized according to the method shown in Example 12 to obtain the title compound as a white powder. Yield 88% ¹ H-NMR (DMSO-d ₆ ): 12.80-13.20 (1H, br), 11.01
(1H, s), 8.62 (1H, d, J = 6.1 Hz), 8.30 (1H, s), 7.8
4 (1H, d, J = 8.0 Hz), 7.74 (1H, d, J = 7.6 Hz), 7.47
(1H, t, J = 7.8 Hz), 7.19-7.28 (5H, m), 6.72 (1H, d,
J = 8.5 Hz), 4.98 (1H, m), 4.34 (1H, m), 3.95 (1H,
m), 3.63 (1H, m), 3.49 (1H, m), 3.16 (1H, dd, J = 14.
2, 4.4 Hz), 2.89 (1H, dd, J = 14.2, 8.5 Hz), 1.77-2.0
0 (5H, m), 1.35 (9H, s), 0.78-0.85 (6H, m) IR (KBr): 3300, 2950, 1690, 1160, 760 cm ^-1 MS (SIMS) m / z 659 (MH ⁺ )

Example 28 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -β-benzyl-L-aspartyl-L-prolyl] amino] -2, 2-difluoro-3
Synthesis of -oxo-5-phenylvaleryl] amino] benzoic acid benzyl ester (Step 1) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-prolyl] Amino] -2,
Synthesis of 2-difluoro-3 (R) -hydroxy-5-phenylvaleryl] amino] benzoic acid benzyl ester N- (tert-butyloxycarbonyl) -L-proline 400 mg (1.86 mmol) of dichloromethane under nitrogen atmosphere. To the solution (15 ml) was added 250 μl of triethylamine, 823 mg (1.86 mmol) of benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate, followed by 250 μl of triethylamine and the purpose of step 2 of Example 26. Compound 400 mg (1.86 mmol) was added. After stirring at room temperature for 40 hours, the reaction product was diluted with 40 ml of dichloromethane, 50 ml of water, 50 ml of 1N hydrochloric acid aqueous solution, 50 ml of saturated aqueous sodium hydrogen carbonate solution, and 50 m of saturated saline.
After washing with l, it was dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by flash column chromatography (hexane: ethyl acetate = 3: 2) to obtain 897 mg of the target compound as a colorless amorphous substance. Yield 81% ¹ H-NMR (DMSO-d ₆ ): 10.67 (1H, s), 8.36 (1H,
s), 7.97 (1H, d, J = 7.9 Hz), 7.78 (1H, d, J = 6.8 Hz),
7.71 (1H, m), 7.22-7.55 (10H, m), 6.48 (1H, d, J =
7.1 Hz), 5.36 (2H, s), 4.27-4.37 (1H, m), 3.99-4.20
(2H, m), 3.19-3.32 (2H, m), 2.73-2.89 (2H, m), 1.6
8-1.90 (4H, m), 1.30 (9H, s) IR (KBr): 3250, 1700, 1660, 1280, 1100, 750, 700
cm ^-1

(Step 2) 3- [N- [4 (S)-[N- (L-prolyl) amino] -2,2-difluoro-3 (R) -hydroxy-5
Synthesis of phenylvaleryl] amino] benzoic acid benzyl ester hydrochloride 847 mg (1.30 mmo of the compound obtained in Step 1
To l), 4N hydrochloric acid (3.3 ml of dioxane solution) was added at 0 ° C. under a nitrogen atmosphere. After stirring at room temperature for 1 hour, the solvent was evaporated under reduced pressure, ether and hexane were added to the residue, and the precipitated powder was collected to obtain 660 mg of the desired compound as a white powder. Yield 87% ¹ H-NMR (DMSO-d ₆ ): 10.69 (1H, s), 9.78 (1H, b
r), 8.62 (1H, d, J = 9.2 Hz), 8.42 (1H, br), 8.37 (1
H, s), 7.99 (1H, d, J = 8.9 Hz), 7.79 (1H, d, J = 7.8 H)
z), 7.19-7.56 (11H, m), 6.65 (1H, d, J = 6.3 Hz), 5.3
7 (2H, s), 4.41 (1H, dd, J = 15.7, 7.7 Hz), 4.16 (2H,
br), 3.15 (2H, m), 2.91 (1H, dd, J = 13.5, 7.3 Hz),
2.79 (1H, dd, J = 13.5, 7.7 Hz), 2.22 (1H, m), 1.83
(3H, m) IR (KBr): 3200, 1700, 1540, 1280, 1100, 750, 700
cm ^-1

(Step 3) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -β-benzyl-L-aspartyl-L-prolyl] amino] -2,2 -Difluoro-3
(R) -Hydroxy-5-phenylvaleryl] amino]
Synthesis of benzoic acid benzyl ester N- (tert-butyloxycarbonyl) -L-aspartic acid β-benzyl ester 12 under nitrogen atmosphere
A solution of 1 mg (0.37 mmol) in dichloromethane (2
50 ml of triethylamine, 165 mg of benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate (0.
37 mmol), followed by triethylamine 50 μ
1 and 200 mg (0.3
4 mmol) was added. After stirring at room temperature for 64 hours, the reaction product was diluted with 20 ml of dichloromethane, and 20 ml of water was added.
After washing with 20 ml of 1N hydrochloric acid aqueous solution, 20 ml of saturated aqueous sodium hydrogen carbonate solution and 20 ml of saturated saline, the organic layer was dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to flash column chromatography (hexane: ethyl acetate = 2:
Purification in 3) yielded 250 mg of the desired compound as a colorless amorphous. Yield 86% ¹ H-NMR (CDCl ₃ ): 9.29 (1H, s), 8.30 (1H, s), 8.
02 (1H, d, J = 8.1 Hz), 7.88 (1H, d, J = 7.8 Hz), 7.16
-7.46 (16H, m), 6.97 (1H, d, J = 8.8 Hz), 5.36 (2H,
s), 5.23 (1H, d, J = 12.1 Hz), 5.16 (1H, d, J = 9.6 H
z), 5.10 (1H, d, J = 12.1 Hz), 4.79 (1H, m), 4.41 (1
H, dd, J = 8.2, 3.3 Hz), 4.26-4.39 (2H, m), 4.00-4.1
8 (1H, m), 3.65 (2H, m), 2.98-3.11 (3H, m), 2.78 (1
H, dd, J = 16.5, 3.8 Hz), 1.80-2.10 (4H, m), 1.43 (9
H, s) IR (KBr): 3300, 2950, 1700, 1640, 1280, 1160, 75
0, 700 cm ^-1

(Step 4) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -β-benzyl-L-aspartyl-L-prolyl] amino] -2,2 -Difluoro-3
Synthesis of -oxo-5-phenylvaleryl] amino] benzoic acid benzyl ester The compound obtained in Step 3 was synthesized according to the method shown in Step 5 of Example 1. For purification, flash column chromatography (hexane: ethyl acetate = 2:
Using 3) the title compound was obtained as a colorless amorphous. Yield 90% ¹ H-NMR (DMSO-d ₆ ): 11.05 (1 H, s), 8.32-8.40 (2
H, m), 7.91 (1H, m), 7.77 (1H, m), 7.17-7.78 (16H,
m), 5.34 (2H, s), 5.05 (2H, s), 4.90-5.10 (1H, m),
4.60 (1H, m), 4.17-4.27 (2H, m), 3.51 (2H, m), 3.19
(1H, m), 2.67-2.86 (2H, m), 2.55 (1H, m), 1.50-2.0
0 (4H, m), 1.34 (9H, s) IR (KBr): 3300, 1700, 1630, 1280, 750, 690 cm ^-1

Example 29 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-aspartyl-L-prolyl] amino] -2,2-difluoro- 3-oxo-5
Synthesis of Phenylvaleryl] amino] benzoic Acid By the method described in Example 12, using the compound obtained in Example 28, the title compound was obtained as a white powder. Yield 7
6% ¹ H-NMR (DMSO-d ₆ ): 12.0-13.3 (2H, br), 10.99
(1H, s), 8.31 (2H, br), 7.86 (1H, d, J = 7.8 Hz), 7.7
4 (1H, d, J = 7.6 Hz), 7.48 (1H, t, J = 7.9 Hz), 7.18-
7.30 (6H, m), 4.97 (1H, m), 4.52 (1H, m), 4.27 (1H,
m), 3.55 (2H, m), 3.20 (1H, dd, J = 14.2, 4.2 Hz),
2.85 (1H, dd, J = 14.0, 9.7 Hz), 2.62 (1H, dd, J = 16.
2, 4.5 Hz), 2.43 (1H, dd, J = 16.2, 8.2 Hz), 1.90 (1
H, m), 1.74 (1H, m), 1.60-1.70 (2H, m), 1.34 (9H,
s) IR (KBr): 3300, 2950, 1700, 1630, 1160, 750 cm
^-1 MS (SIMS) m / z 675 (MH ⁺ )

Example 30 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -γ-benzyl-L-glutamyl-L-prolyl] amino] -2, 2-difluoro-3-
Synthesis of oxo-5-phenylvaleryl] amino] benzoic acid benzyl ester (Step 1) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -γ-benzyl-L -Glutamyl-L-prolyl] amino] -2,2-difluoro-3
(R) -Hydroxy-5-phenylvaleryl] amino]
Synthesis of benzoic acid benzyl ester N- (tert-butyloxycarbonyl) -L-aspartic acid N- (t
ert-Butyloxycarbonyl) -L-glutamic acid γ-benzyl ester, and was synthesized by the method shown in Step 3 of Example 28. For purification, flash column chromatography (hexane: ethyl acetate =
2: 3) was used to obtain the target compound as a colorless amorphous. Yield 76% ¹ H-NMR (CDCl ₃ ): 9.20 (1H, br), 8.32 (1H, br),
7.99 (1H, d, J = 8.7 Hz), 7.87 (1H, d, J = 7.2 Hz), 7.
14-7.44 (16H, m), 6.97 (1H, d, J = 8.1 Hz), 5.35 (2H,
s), 5.32 (1H, d, J = 7.9 Hz), 5.12 (2H, s), 4.94 (1
H, m), 4.10-4.62 (4H, m), 3.61 (2H, m), 2.93-3.10
(2H, m), 2.51 (2H, m), 1.70-2.20 (6H, m), 1.43 (9
H, s) IR (KBr): 3300, 1700, 1280, 1160, 750, 700 cm ^-1

(Step 2) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -γ-benzyl-L-glutamyl-L-prolyl] amino] -2,2 -Difluoro-3-
Synthesis of oxo-5-phenylvaleryl] amino] benzoic acid benzyl ester The target compound obtained in Step 1 was synthesized according to the method shown in Step 5 of Example 1. For purification, flash column chromatography (hexane: ethyl acetate =
2: 3) was used to give the title compound as a colorless amorphous. Yield 75% ¹ H-NMR (DMSO-d ₆ ): 11.04 (1H, s), 8.56 (1H, d,)
J = 7.2 Hz), 8.33 (1H, br), 7.93 (1H, d, J = 8.0 Hz),
7.79 (1H, d, J = 7.8 Hz), 7.16-7.52 (16H, m), 6.97 (1
H, d, J = 8.1 Hz), 5.35 (2H, s), 5.07 (2H, s), 4.97
(1H, m), 4.29 (1H, m), 4.19 (1H, m), 3.45 (2H, m),
3.17 (1H, dd, J = 14.4, 5.0 Hz), 2.85 (1H, dd, J = 14.
4, 8.9 Hz), 2.40 (2H, m), 1.60-2.00 (6H, m), 1.34
(9H, s) IR (KBr): 3300, 1700, 1630, 1160, 750, 700 cm ^-1

(Example 31) 3- [N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-glutamyl-L-prolyl] amino] -2,2-difluoro- 3-oxo-5
Synthesis of Phenylvaleryl] amino] benzoic Acid By the method described in Example 12, using the compound obtained in Example 30, the title compound was obtained as a white powder. Yield 7
4% ¹ H-NMR (DMSO-d ₆ ): 12.2-13.2 (2H, br), 10.99
(1H, s), 8.58 (1H, d, J = 7.2 Hz), 8.31 (1H, s), 7.85
(1H, d, J = 8.2 Hz), 7.74 (1H, d, J = 7.8 Hz), 7.47
(1H, t, J = 7.9 Hz), 7.19-7.29 (5H, m), 6.95 (1H, d,
J = 8.0 Hz), 4.97 (1H, m), 4.31 (1H, m), 4.19 (1H,
m), 3.54 (2H, m), 3.19 (1H, dd, J = 14.2, 4.5 Hz), 2.8
6 (1H, dd, J = 14.2, 8.9 Hz), 2.27 (1H, t, J = 7.0 H
z), 1.50-2.00 (6H, m), 1.35 (9H, s) IR (KBr): 3300, 1700, 1630, 1160 cm ^-1 MS (SIMS) m / z 689 (MH ⁺ )

Example 32 3- [N- [4 (S)-[N- [N- (phenylsulfonyl) -L-glutamyl-L-prolyl] amino]-
Synthesis of 2,2-difluoro-3-oxo-5-phenylvaleryl] amino] benzoic acid (Step 1) 3- [N- [4 (S)-[N- [L-γ-benzylglutamyl-L- Synthesis of prolyl] amino] -2,2-difluoro-3 (R) -hydroxy-5-phenylvaleryl] amino] benzoic acid benzyl ester hydrochloride 300 mg (0.34) of the target compound in Step 1 of Example 30
(4 mmol) (dioxane solution 1.3 ml) was added to the (mmol) at 0 ° C. under a nitrogen atmosphere. After stirring at room temperature for 16 hours, the solvent was evaporated under reduced pressure, ether was added to the residue, and the precipitated powder was collected to obtain 260 mg of the desired compound as a white powder. Yield 94% ¹ H-NMR (DMSO-d ₆ ): 10.63 (1H, s), 8.36 (1H,
s), 8.26 (3H, br), 8.02 (1H, d, J = 9.1 Hz), 7.96 (1
H, d, J = 8.6 Hz), 7.78 (1H, m), 7.53-7.15 (15H, m),
6.49 (1H, d, J = 7.3 Hz), 5.36 (2H, s), 5.11 (2H,
s), 4.53 (11H, m), 4.30 (1H, m), 4.18 (1H, t, J = 6.1
Hz), 4.05 (1H, m), 3.60 (1H, m), 3.43 (1H, m), 2.8
5 (1H, m), 2.75-2.50 (3H, m), 2.10-1.65 (6H, m) IR (KBr): 3250, 1720, 1640, 1280, 750, 700 cm ^-1

(Step 2) 3- [N- [4 (S)-[N- [N- (phenylsulfonyl) -γ-benzyl-L-glutamyl-L-prolyl] amino] -2,2-difluoro- Synthesis of 3 (R) -hydroxy-5-phenylvaleryl] amino] benzoic acid benzyl ester 250 mg of the target compound obtained in Step 1 under nitrogen atmosphere
A solution of (0.30 mmol) in dichloromethane (2 ml)
Triethylamine 91 μl (0.65 mmol) and benzenesulfonyl chloride 60 mg (0.34 mmo)
1) was added, and the mixture was stirred at room temperature for 16 hours. The reaction solution was diluted with 30 ml of dichloromethane, and the aqueous solution of 1N hydrochloric acid was 30 m.
1, washed with 30 ml of saturated aqueous sodium hydrogen carbonate solution and 30 ml of saturated saline solution, and dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by flash column chromatography (hexane: ethyl acetate = 1: 2), 20
0 mg of the desired compound was obtained as a colorless amorphous. Yield 71% ¹ H-NMR (CDCl ₃ ): 9.08 (1H, s), 8.25 (1H, s), 7.
97 (1H, d, J = 7.0 Hz), 7.78 (1H, d, J = 7.9 Hz), 7.80
-7.76 (2H, m), 7.50-7.11 (19H, m), 6.80 (1H, d, J =
8.2 Hz), 5.83 (1H, d, J = 9.7 Hz), 5.36 (2H, s), 5.13
(1H, d, J = 12.2 Hz), 5.07 (1H, d, J = 12.2 Hz), 4.73
(1H, d, J = 7.3 Hz), 4.18-4.02 (3H, m), 3.88 (1H,
m), 3.47 (1H, m), 3.10-2.94 (3H, m), 2.68 (1H, m),
2.46 (1H, m), 1.96-1.40 (6H, m) IR (KBr): 3300, 1710, 1280, 1160, 750, 695 cm ^-1

(Step 3) 3- [N- [4 (S)-[N- [N- (phenylsulfonyl) -γ-benzyl-L-glutamyl-L-prolyl] amino] -2,2-difluoro- 3-oxo-5
Synthesis of phenylvaleryl] amino] benzoic acid benzyl ester 186 mg (0.2 mmo of the target compound obtained in Step 2)
l) was synthesized according to the method shown in Step 5 of Example 1 to obtain 158 m of the target compound as colorless amorphous.
g was obtained. Yield 85% ¹ H-NMR (DMSO-d ₆ ): 10.99 (1H, s), 8.49 (1H, d,)
J = 7.3 Hz), 8.32 (1H, s), 8.08 (1H, d, J = 9.2 Hz),
7.90 (1H, m), 7.78 (1H, d, J = 6.4 Hz), 7.72-7.68 (2
H, m), 7.60-7.10 (19H, m), 5.35 (2H, s), 5.06 (1H,
d, J = 12.5 Hz), 5.02 (1H, d, J = 12.5 Hz), 4.92 (1H,
m), 3.99 (1H, m), 3.82-3.73 (1H, m), 3.19-3.10 (2
H, m), 2.79 (1H, dd, J = 14.2, 8.9 Hz), 2.36 (2H, m),
1.90-1.45 (6H, m), IR (KBr): 3250, 1720, 1280, 1160, 750, 690 cm ^-1

(Step 4) 3- [N- [4 (S)-[N- [N- (phenylsulfonyl) -L-glutamyl-L-prolyl] amino]-
Synthesis of 2,2-difluoro-3-oxo-5-phenylvaleryl] amino] benzoic acid 145 mg (0.16 mm) of the target compound obtained in Step 3
ol) according to the method described in Example 12 to give 88 mg of the title compound as a colorless amorphous. Yield 77% ¹ H-NMR (DMSO-d ₆ ): 13.5-12.0 (2H, br), 10.92
(1H, s), 8.47 (1H, d, J = 7.3 Hz), 8.29 (1H, s), 8.02
(1H, d, J = 9.3 Hz), 7.82 (1H, d, J = 8.6 Hz), 7.76-7.
69 (3H, m), 7.62-7.47 (4H, m), 7.29-7.14 (5H, m),
4.93 (1H, m), 4.01 (1H, m), 3.97 (1H, m), 3.47 (2H,
m), 3.15 (1H, dd, J = 14.1, 4.7 Hz), 2.81 (1H, dd, J
= 14.1, 8.9 Hz), 2.24 (2H, t, J = 7.4 Hz), 1.90-1.50
(6H, m) IR (KBr): 3250, 1700, 1630, 1160, 750 cm ^-1 MS (SIMS) m / z 729 (MH ⁺ )

The compounds obtained in Examples 26 to 32 are summarized in Table 3. In addition, Compounds 33 to 54 produced by the same method as in the above Examples are also shown in Tables 3 and 4. However, Examples 35 to 3 in Table 3 and Table 4
For 7 of the compound, R _5, X is expressed as -N (R ₅₎ -X. Ala, Asp, Glu and Bzl are
They represent alanine, aspartic acid, glutamic acid and benzyl, respectively.

[0150]

[Table 3]

[0151]

[Table 4]

Reference Example N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-methylamine (Step 1) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl)- L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3 (R) -hydroxy-5-phenylvaleryl] -N-methylamine Synthesized by the method shown in Step 4 of Example 1 using methylamine instead of benzylamine. . For purification, flash column chromatography (hexane:
The target compound was obtained as a colorless amorphous using ethyl acetate = 2: 3). Yield 95% IR (KBr): 3300, 2950, 1680, 1630, 1160 cm ^-1

(Step 2) N- [4 (S)-[N- [N- (tert-butyloxycarbonyl) -L-valyl-L-prolyl] amino]
Synthesis of -2,2-difluoro-3-oxo-5-phenylvaleryl] -N-methylamine The compound obtained in Step 1 was synthesized according to the method shown in Step 5 of Example 1, and the title compound was white. Obtained as an amorphous. Yield 60% ¹ H-NMR (DMSO-d ₆ ): δ 9.08 (1H, br), 8.52 (1H,
d, J = 6.8 Hz), 7.26-7.37 (5H, m), 6.80 (1H, d, J = 8.7
Hz), 4.98 (1H, br), 4.2-4.5 (2H, m), 3.8-4.1 (1H,
m), 3.65-3.8 (1H, m), 3.4-3.65 (1H, m), 3.22 (1H, d
d, J = 14.7, 4.5 Hz), 2.88 (1H, dd, J = 14.7, 8.9 Hz),
2.75 (3H, m), 1.6-2.2 (5H, m), 1.42 (9H, s), 0.93
(3H, d, J = 6.7 Hz), 0.89 (3H, d, J = 6.3 Hz) IR (KBr): 3300, 2950, 1680, 1620, 1500, 1160 cm
^-1 MS (SIMS) m / z 553 (MH ⁺ )

The following activities were evaluated for the compounds obtained in the examples. Experimental Example 1: Inhibitory activity of the compound of the present invention on human cardiac chymase The inhibitory activity of the compound (I) of the present invention was measured by the inhibitory activity on the amidase activity of human cardiac chymase, and its effectiveness was evaluated as follows.

Synthetic substrate succinyl-at a final concentration of 2.5 mM
Alanyl-alanyl-prolyl-phenylalanine-p
The inhibitory activity was quantified by the change in the residual activity fraction by the compound of the present invention in the specified concentration series (<× 1, <× 10, <× 100 times equivalent) against 5 nM chymase in the presence of -nitroanilide. Inhibition potency was analyzed by Easson-Stedman plot (Proc. Roy.
Soc. B. 1936, 121, p141). The apparent inhibition constant (Kiapp) obtained from this analysis,
The inhibitory activity was evaluated by the inhibitory constant (Ki) calculated from the final concentration of the reaction solution substrate and the separately determined Km value. The initial rate of the enzymatic reaction was quantified by spectrophotometrically detecting the amount of p-nitroaniline produced by hydrolysis of the substrate by measuring the increase in absorbance at 405 nm. The chymase inhibitory activity of the compound of the present invention is calculated as an activity remaining fraction in the presence of the inhibitor with respect to the enzyme activity in the absence of the inhibitor, and the measured value is less than the initial rate guaranteed absorbance at the substrate concentration used for the enzyme. After incorporation, analysis was performed.

The reaction solution composition was 0.1% Triton-.
The composition including X100 is Na ₂ B ₄ O ₇ (100 mM)
-KH ₂ PO ₄ (50mM) in a buffer (pH 9.
0) 120 μl of the compound of the present invention dissolved in 20 μl of 10% dimethyl sulfoxide (DMSO), 20 μl of bovine serum albumin 10% dissolved in the same buffer, 20 μl
Substrate dissolved in DMSO and 20 μl of chymase were added to make a total volume of 200 μl. From the absorbance immediately after addition of the enzyme, the increase in absorbance was recorded as a progressive curve at exactly equal time intervals. From the above data, if necessary, the difference in the absorbance immediately after addition from the absorbance at the end of the reaction, the residual activity of the inhibitor-added sample relative to the control without the inhibitor was quantified and analyzed, or for a certain period of time. The reaction rates of the control and inhibitor-added samples were calculated with a step size (≧ 20 minutes), the rate calculation was shifted every 10 to 30 minutes, and the same reaction rate was averaged over the entire reaction time. The inhibitory activity was analyzed by a method of quantifying the residual activity fraction.

In addition, the inhibitory activity against bovine chymotrypsin was measured using N-methoxysuccinyl-arginyl-prolyl-tyrosine-p-nitroanilide as a substrate and 20 mM CaCl ₂ and 0.1% as a buffer.
0.1 M Tris-hydrochloric acid (pH 8.0) containing Triton-X100 was used, and otherwise the composition and method were the same as described above. Since bovine chymotrypsin and human chymotrypsin have high homology, it is presumed that similar results are obtained with human chymotrypsin.

Table 5 shows the results of the above-described activity measurement test for the representative compound (I) of the present invention.

[0159]

[Table 5]

From the above results, it is understood that the compound (I) of the present invention inhibits human cardiac chymase much more potently than the inhibition of bovine chymotrypsin.

Formulation Example 1: Tablet (1) Compound (I) of the present invention 10 mg (2) Fine granules for direct compression No. 209 (manufactured by Fuji Chemical Co., Ltd.) 46.6 mg Magnesium aluminometasilicate 20% Corn starch 30% Lactose 50% (3) Crystalline cellulose 24.0 mg (4) Carboxymethyl cellulose / calcium 4.0 mg (5) Magnesium stearate 0. 4 mg (1), (3) and (4) are all passed through a 100 mesh sieve in advance. These (1), (3), (4) and (2)
Each of them was dried to reduce the water content to a constant value, and then mixed in the above weight ratio using a mixer. (5) was added to the mixed powder which was made uniform in quality and mixed for a short time (30 seconds), and the mixed powder was tableted (pestle: 6.3 mmφ, 6.0 mmR),
One tablet was 85 mg. This tablet is a gastric-soluble film coating agent that is usually used as needed (eg, polyvinyl acetal diethylaminoacetate).
It may also be coated with an edible coloring agent.

Formulation Example 2: Capsule (1) Compound (I) of the present invention 50 g (2) Lactose 935 g (3) Magnesium stearate 15 g The above components were weighed and mixed uniformly, and the mixed powder was hard gelatin capsules. Was filled with 200 mg each.

Production Example 3: Injection (1) Hydrochloride of the compound (I) of the present invention 5 mg (2) Sucrose 100 mg (3) Physiological saline 10 ml The above mixed solution was filtered through a membrane filter and then sterilized again. The filtrate was aseptically dispensed into a vial, filled with nitrogen gas, and then sealed to give an intravenous injection.

[0164]

The peptide compound of the present invention and the pharmacologically acceptable salt thereof are suitable for mammals including humans.
It has an excellent inhibitory effect on the chymase group and / or has high selectivity for the chymase group. Therefore, it is useful as a chymase inhibitor, and is useful for the prevention / treatment of various diseases caused by chymase including various diseases caused by angiotensin II.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical indication location A61K 38/55 ADP A61K 37/64 ADP AEQ AEQ (72) Inventor Yoshihisa Inoue Invited Otani, Hirakata, Osaka 2-25-1 Company Midori Cross Central Research Institute (72) Inventor Hajime Fukuyama Osaka Prefecture Hirakata City Invited Otani 2-25-1 Company Midori Cross Central Research Institute (72) Inventor Nakajima Masahide Osaka Prefecture Hirakata City Invited 2-25-1 Otani Co., Ltd. in the Midori Cross Central Research Institute (72) Inventor Mitsuaki Imada Osaka Prefecture Hirakata Invited 2-25-1 Otani Incorporated Central Research Institute (72) Inventor Hideki Okunishi 5-11-5 Tsukiwa, Otsu City, Shiga Prefecture (72) Inventor Miya ▲ Saki ▼ Mizuo 4-5 Tomooka, Nagaokakyo, Kyoto Prefecture

Claims (8)

[Claims] 1. A compound of the formula (I) [In the formula, X is Or (Wherein, Z _1, Z ₂ independently represent hydrogen, hydroxyl, a group represented by -COR _8, alkyl optionally substituted with a group represented by -COR ₈ or -S,
Represents a group represented by O ₂ R ₈ , R ₆ and R ₇ are the same or different and are hydrogen, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, -C
Alkyl, alkenyl, alkynyl substituted with a group represented by OR ₈ or a group represented by —COR ₈ ;
Represents a heterocycle or heterocycle alkyl,
However, R ₆ and R ₇ do not represent hydrogen at the same time, and R ₆ and R 7
_When either one or both of ₇ is a heterocycle substituted with a group represented by —COR ₈ , the heteroatom of the heterocycle does not bond to a carbon atom bonded to R ₆ or R ₇ , or R ₆ and R ₇ together-
C 4 may represent a methylene chain having 4 to 6 carbon atoms which may be substituted with a group represented by COR ₈ , and ring B represents a group represented by an aromatic ring), and R ₁ represents —COR ₈ Represents an alkyl, cycloalkyl, cycloalkylalkyl, alkoxy, aryl, arylalkyl, heteroaryl, heterocycle or heterocyclealkyl which may be substituted with a group represented by R ₂ , R ₃ are the same or different. Hydrogen, or -COR
_An alkyl optionally substituted with a group represented by ₈ ,
Cycloalkyl, cycloalkylalkyl, aryl or arylalkyl is shown, or R ₂ and R ₃ together form a methylene chain having 4 to 6 carbon atoms which may be substituted with a group represented by —COR _8. R ₄ may be hydrogen or alkyl, R ₅ may be hydrogen or alkyl, or X may be In the case of, R ₆ may represent a methylene chain having 2 to 4 carbon atoms together with R _6, and R ₈ is a hydroxyl group, alkoxy, amino, alkylamino,
Dialkylamino, aryloxy or arylalkoxy, Y represents cycloalkyl, aryl or heteroaryl, m represents an integer of 0 or 1 to 3, and A represents carbonyl or sulfonyl] Alternatively, a pharmacologically acceptable salt thereof. 2. In the formula (I), X is (Wherein, Z _1, Z ₂ independently represent hydrogen, hydroxyl, a group represented by -COR _8, alkyl optionally substituted with a group represented by -COR ₈ or -S,
O ₂ R ₈ represents a group), R ₁ represents alkyl, R ₂ and R ₃ are the same or different and are hydrogen or —COR ₈
Represents an alkyl which may be substituted with a group represented by R ₄ , R ₄ represents hydrogen, R ₅ represents hydrogen or alkyl, R ₈ represents a hydroxyl group, alkoxy or arylalkoxy, Y represents aryl, The peptide compound according to claim 1, wherein m represents 1, and A represents carbonyl or sulfonyl, or a pharmacologically acceptable salt thereof. 3. At least one of Z ₁ and Z ₂ is --CO
The peptide compound according to claim 2, which is alkyl substituted with a group represented by R ₈ or a group represented by —COR ₈ , or a pharmacologically acceptable salt thereof. 4. In the formula (I), X is (Wherein, R _6, R ₇ represent the same or different and each is hydrogen, aryl, arylalkyl, an alkyl substituted with a group represented by the group or -COR ₈ represented by -COR _8, where R _6, R ₇ does not represent hydrogen at the same time), R ₁ represents alkyl, R ₂ and R ₃ are the same or different and are hydrogen or —COR ₈
Represents an optionally substituted alkyl group, R ₄ represents hydrogen, R ₅ represents hydrogen or alkyl, or R ₆ represents a methylene chain having 3 carbon atoms. Often, R ₈ represents a hydroxyl group, alkoxy or arylalkoxy, Y represents aryl, m represents 1, and A represents carbonyl or sulfonyl, and the peptide compound according to claim 1 or a pharmacologically acceptable thereof. salt. 5. At least one of R ₆ and R ₇ is —COR.
Group or a peptide compound or a pharmacologically acceptable salt thereof according to claim 4 represents an alkyl substituted with a group represented by -COR ₈ is represented by _8. 6. Formula (II): [In the formula, X is Or (Wherein, Z _1, Z ₂ independently represent hydrogen, hydroxyl, a group represented by -COR _8, alkyl optionally substituted with a group represented by -COR ₈ or -S,
Represents a group represented by O ₂ R ₈ , R ₆ and R ₇ are the same or different and are hydrogen, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, -C
Alkyl, alkenyl, alkynyl substituted with a group represented by OR ₈ or a group represented by —COR ₈ ;
Represents a heterocycle or heterocycle alkyl,
However, R ₆ and R ₇ do not represent hydrogen at the same time, and R ₆ and R 7
_When either one or both of ₇ is a heterocycle substituted with a group represented by —COR ₈ , the heteroatom of the heterocycle does not bond to a carbon atom bonded to R ₆ or R ₇ , or R ₆ and R ₇ together-
C 4 may represent a methylene chain having 4 to 6 carbon atoms which may be substituted with a group represented by COR ₈ , B ring represents a group represented by), and R ₂ and R ₃ represent Identical or different hydrogen, or -COR
_An alkyl optionally substituted with a group represented by ₈ ,
Cycloalkyl, cycloalkylalkyl, aryl or arylalkyl is shown, or R ₂ and R ₃ together form a methylene chain having 4 to 6 carbon atoms which may be substituted with a group represented by —COR _8. R ₄ may be hydrogen or alkyl, R ₅ may be hydrogen or alkyl, or X may be In the case of, R ₆ may represent a methylene chain having 2 to 4 carbon atoms together with R _6, and R ₈ is a hydroxyl group, alkoxy, amino, alkylamino,
Represents a dialkylamino, aryloxy or arylalkoxy, Rp represents a protecting group for an amino group, Y represents cycloalkyl, aryl or heteroaryl, m represents an integer of 0 or 1 to 3, and A represents carbonyl or And a pharmacologically acceptable salt thereof. 7. A pharmaceutical composition comprising the peptide compound according to claim 1 or 2, or a pharmacologically acceptable salt thereof. 8. The pharmaceutical composition according to claim 4, which is a chymase inhibitor.