JPH05201970A - New proline derivative - Google Patents

Abstract

(57) [Summary] (Modified) [Structure] General formula [A represents -O-, -NH-, -CONH- or a single bond;
X is -S -, - SO -, - SO 2 -, - O- or -NH
-Is; Y and Z are the same or different and each is H, halogen, a lower alkyl group which may be substituted with F, an amino group, a nitro group,
A hydroxyl group, a lower alkoxy group, and a saturated or unsaturated 5- and 6-membered ring in which Y and Z are combined; R ² is H, a halogen or a lower alkoxy group; n is an integer of 1 to 6; 0
Represents an integer of ~ 3]. [Effects] Proline derivatives have a strong inhibitory activity specifically on prolyl endopeptidase, suppress the degradation and inactivation of TRH, substance P, neurotensin, vasopressin, etc. It can be used as an amnestic drug for child defense and / or treatment.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a novel proline derivative having prolyl endopeptidase inhibitory activity and is used in the field of medicine.

[0002]

2. Description of the Related Art With the advent of an aging society, the problem of medical care for the elderly has been emphasized, and senile dementia has become a serious social problem. In order to cope with this, various developments of new medicines have been made. However, there are many ambiguous expressions such as cerebral circulation improving agents, cerebral metabolism activating agents or cerebral function improving agents due to their mechanism of action in conventional therapeutic agents for amnesia, dementia, etc. Although it is effective in improving peripheral symptoms, its effect is not always clear for core symptoms of dementia such as memory impairment and disorientation, and the development of a drug that brings more reliable action is desired. There is.

On the other hand, prolyl endopeptidase (Pr
oryl endopeptidase; EC, 3.
4.21.26) is an enzyme that acts on a peptide containing proline, and is known to specifically cleave the carboxyl group side of proline. This enzyme acts on neurotransmitters such as thyrotropin-releasing hormone (TRH), substance P, and neurotensin, as well as learning,
It is also known to act on vasopressin, which is said to be involved in memory processes, to decompose and inactivate them.

From these findings, if compounds having an inhibitory activity specific to the present enzyme can be obtained, these compounds suppress the decomposition and inactivation of vasopressin and the like,
As a drug that directly acts on the core symptoms of dementia, it is expected to be applicable to the prevention and treatment of amnesia or dementia (Biochemistry, 55 , 831 (1983): Jpn Jpn Pharmacol. 89 , 24.
3 (1987): J. Pharmacobio-Dy
n. , 10 , 730 (1987)), and TRH,
By suppressing the degradation and inactivation of substance P, hormones such as neurotensin, and neurotransmitters, it is expected to be effective in improving the symptoms of diseases caused by the degradation and inactivation of these substances. To be done. Recently
In vitro and in vivo experiments, it has been shown that beta-amyloid protein plays an essentially important role in the development of Alzheimer's disease by exhibiting a neurotoxic effect. Hypothesis that prolyl endopeptidase is a beta amyloid excision enzyme from amyloid precursor protein (FEBS Lett., 260, 131
-134 (1990)), or experimental fact that the neurotoxic effect of beta amyloid protein is suppressed by substance P (Proc. Natl. Aca.
d. Sci. USA, 88, 7247-7251 (19
From 91)), it is considered that a prolyl endopeptidase inhibitor can be an effective therapeutic agent for Alzheimer's disease. Various developments relating to prolyl endopeptidase inhibitors have been made in the past. For example, JP-A-62-14
Various proline derivatives are described and disclosed in, for example, 8467, JP-A-64-42475, JP-A-64-6263, JP-A-1-230578 or JP-A-2-28149. There is.

[0005]

Therefore, the present inventors have found that
Based on the above findings, as a result of intensive research to find a compound having an amino acid, particularly a proline residue, as a fragment and specifically and strongly inhibiting the action of prolyl endopeptidase, the result of the above general formula [1] It was found that the proline derivative shown has a specific and strong prolyl endopeptidase inhibitory activity, and completed the present invention.

[0006]

That is, the present invention provides the following general formula [1]

[Chemical 5] [In the formula, A is -O-, -NH-, -CONH- or a single bond, and B is

[Chemical 6] - _{(CH 2) K} - or -NHCHR ¹ - a, (however, k
Is an integer of 1 to 3, R ¹ is a hydrogen atom or a lower alkyl group), and W is

[Chemical 7] Or CH ₃ — (wherein R ² represents a hydrogen atom, a halogen atom or a lower alkoxy group), X is —S—, —S.
_{O -, - SO 2 -,} - O- or -NH- and R is

[Chemical 8] Or a lower alkyl group (wherein 1 is an integer of 0 to 3,
Y and Z are the same or different and each is a hydrogen atom, a halogen atom,
It means a lower alkyl group which may be substituted with a fluorine atom, an amino group, a nitro group, a hydroxyl group or a lower alkoxy group.
Y and Z may be combined to form a saturated or unsaturated 5- and 6-membered ring), and n is an integer of 1 to 6]. Another object of the present invention is to provide a pharmaceutical composition useful as a prolyl endopeptidase inhibitor containing the novel proline derivative [1] as an active ingredient.

The definitions of various substituents used in the present specification are as follows. The "lower alkyl group" means a straight or branched hydrocarbon chain having 1 to 5 carbon atoms, and specifically, methyl group, ethyl group, propyl group, isopropyl group, butyl group, sec-butyl group. , Tert-
Examples thereof include a butyl group and a pentyl group. "Lower alkoxy group"
Is an alkoxy group having 1 to 5 carbon atoms, specifically, methoxy group, ethoxy group, propoxy group, isopropoxy group, butoxy group, sec-butoxy group, tert-
Butoxy group and the like. The "halogen atom" is chlorine, bromine, fluorine or iodine. The “alkyl group which may be substituted with a fluorine atom” means the above-mentioned C 1 to C 5
Is a lower alkyl group, which may be substituted with a fluorine atom, specifically, a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, a sec-butyl group,
A tert-butyl group, a pentyl group, a difluoromethyl group, a difluoroethyl group, a trifluoromethyl group, a trifluoroethyl group and the like. "Y and Z together are a saturated or unsaturated five- or six-membered ring" means a furan ring, an oxolane ring, a 1,3-dioxolane ring, a thiophene ring, a pyrrole ring, a pyrrolidine ring, an oxane ring, a pyridine ring. , Benzene ring and the like. The novel proline derivative represented by the general formula [1] according to the present invention can be produced, for example, according to the reaction steps shown below.

[0008]

[Chemical 9]

[Chemical 10] Each of the above reaction steps will be further described. In addition,
The symbols A, B, W, X, R, k and n have the same meanings as described above.

Reaction (A) The compound represented by the general formula [3] is reacted with a sulfur ylide derived from trimethylsulfonium iodide or trimethylsulfoxonium iodide in the presence of a strong base. To obtain the epoxide represented by. This reaction uses, for example, n-butyllithium or sodium hydride-dimethylsulfoxide in an anhydrous inert solvent such as tetrahydrofuran, 1,4-dioxane, hexane to give the sulfaylide from the above sulfonium salt and then This is achieved by reacting with compound [2]. The reaction temperature is -70 ° C to reflux temperature, preferably -10 ° C to room temperature. Also,
The compound [3] can also be obtained by subjecting the compound [2] to an olefin by subjecting it to a Wittig reaction and then epoxidizing it with a peracid. Specifically, for example,
The olefin is obtained by forming the corresponding ylide from methyltriphenylphosphonium halide with n-butyllithium or the like in an inert solvent such as tetrahydrofuran or diethyl ether, and then reacting it with compound [2]. The reaction temperature is appropriately from -70 ° C to the reflux temperature. This olefin is used in a solvent such as methylene chloride, benzene, hexane, and methanol at -20 ° C to reflux temperature, preferably 0 ° C to room temperature, using an organic catalyst such as m-chloroperbenzoic acid or hydrogen peroxide solution. The desired compound [3] can be obtained by carrying out the epoxidation reaction.

Reaction (B) The compound represented by the general formula [3] can be prepared by reacting the compound represented by the general formula HX-R (X) in the presence or absence of a base in a suitable solvent.
= S, O, NH) and the compound [4]
The alcohol body represented by the general formula [5] is obtained. Specifically, for example, the reaction with HS-R in which X is a sulfur atom is carried out in the presence of a tertiary amine such as triethylamine or N-methylmorpholine in the presence of methanol,
It is carried out in a solvent such as 1,4-dioxane or N, N-dimethylformamide. HO- in which X is an oxygen atom
In the case of reaction with R, either with sodium methoxide in methanol, or 1,4-dioxane, N, the HO-R with a solvent sodium hydride such as N- dimethylformamide anion ^- O-R After that, the reaction with [3] is performed. The reaction with H ₂ N—R where X is NH is
It is carried out in a solvent such as methanol or 1,4-dioxane. The reaction temperature is room temperature to reflux temperature in each case.

Reaction (C) General formula [5] or

The t-butoxycarbonyl group (Boc group) which is the amino-protecting group of the intermediate represented by [9] is removed according to a known method, and this is subjected to a condensation reaction with the compound represented by the formula [6] to obtain a compound [7] and [1
0] is obtained. Removal of the Boc group, which is an amino protecting group, can be carried out by using the formula [5]

The intermediate represented by [9] is prepared by a known method using hydrobromic acid / acetic acid, hydrochloric acid / dioxane, formic acid, hydrochloric acid / acetic acid, trifluoroacetic acid or the like at −30 ° C.
It can be removed by acid treatment at a temperature of from 70 to 70 ° C, preferably from 0 to 30 ° C. Next, the B removal obtained in this way
The oc form is subjected to a condensation reaction with the compound [6] by a conventional method to obtain the amino acid derivative [7] or [10]. For this peptide formation reaction, a method known per se can be adopted. As a method which can be usually used, N, N′-dicyclohexylcarbodiimide (DCC), water-soluble carbodiimide hydrochloride (1-ethyl-3- (3-dimethylaminopropyl))
Examples thereof include a method using carbodiimide hydrochloride (EDC · HCl) as a condensing agent, an active ester method, a mixed acid anhydride method, and the like. The reaction is carried out in an inert solvent at 0 ° C to under heating. Suitable solvents include chloroform, diethyl ether,
N, N-dimethylformamide, ethyl acetate, dichloromethane, tetrahydrofuran or the like can be used.
The active ester method is carried out by reacting the above compound [6] with p-nitrophenol, thiophenol, p-nitrothiophenol, N-hydroxysuccinimide, etc. in the presence of DCC in an inert solvent to give an active ester (for example, , An ester with N-hydroxysuccinimide), which is isolated or is not isolated, and is further reacted with the de-Boc derivative at 0 ° C. to 40 ° C. in an inert solvent to form a peptide bond. Is. The mixed acid anhydride method is carried out in the presence of a tertiary amine (for example, pyridine or triethylamine) in an inert solvent in the presence of the above compound [6].
And acid halides (eg pivaloyl chloride, tosyl chloride, oxalyl chloride) or acid derivatives (eg ethyl chloroformate, isobutyl chloroformate).
A mixed acid anhydride is obtained by reacting at 0 ° C to 40 ° C, and the mixed acid anhydride is further reacted with the de-Boc derivative at 0 ° C to 40 ° C to form a peptide bond. Further, the DCC method is carried out in the presence or absence of the tertiary amine such as triethylamine in an inert solvent, or a suitable additive (for example, 1-hydroxybenzotriazole (HOBt), N-hydroxy-5-). Norbornene-2,3-dicarboxylic acid imide (HON
Under the conditions of addition or non-addition of B)), a desired peptide bond is formed by reacting the de-Boc derivative with [6] using DCC, EDC.HCl or the like as a condensing agent.

Reaction (D) The final object [1] is obtained by oxidizing the alcohol compound represented by the general formula [7], [8] or [11] with an appropriate oxidizing agent. This reaction is carried out in an inert solvent such as benzene, methylene chloride or N, N-dimethylformamide at 0 ° C to reflux temperature, preferably 0 ° C to room temperature in the presence or absence of molecular sieves. Underneath, pyridinium chlorochromate or pyridinium dichromate is used, or in an inert solvent such as methylene chloride, from -80 ° C to room temperature, preferably -8.
Pyridine, trifluoroacetic acid, at 0 ° C. to 0 ° C., in the presence of oxalyl chloride and triethylamine, using dimethyl sulfoxide, or in the presence or absence of an inert solvent (for example, benzene) at 0 ° C. to room temperature, In the presence of dimethyl sulfoxide, DCC is used, or in the presence or absence of an inert solvent (eg, benzene) -1
This is accomplished by using the sulfur trioxide-pyridine complex in the presence of a tertiary amine such as triethylamine and dimethylsulfoxide at 0 ° C to room temperature.

Reaction (E) Of the compounds represented by the general formula [7], a compound in which X is a sulfur atom is oxidized with a peracid such as m-chloroperbenzoic acid to give the compound represented by the general formula [8]. To obtain a sulfoxide form and a sulfone form. Specifically, in an inert solvent such as methylene chloride, chloroform, benzene, -2
By using 1 equivalent or 2 equivalents of m-chloroperbenzoic acid at 0 ° C to reflux temperature, preferably 0 ° C to room temperature, a sulfoxide compound or a sulfone compound can be obtained.

Reaction (F) The compound represented by the general formula [5] in which X is NH is subjected to an amino protecting group R ^{3 by} a method known in peptide chemistry.
Introduce the compound

[9] is obtained. Although various kinds of amino protecting groups R ³ can be considered, the following reaction (

[9] → [10]) must be stable against acid treatment for de-Boc, and examples thereof include acyl-type protecting groups such as formyl group and trifluoroacetyl group, and urethane-type protecting groups. -Fluorenylmethyloxycarbonyl group, methylsulfonylethyloxycarbonyl group and the like. The introduction of these protecting groups can be achieved by a known method (“Basics and Experiments of Peptide Synthesis”, Izumiya et al., Maruzen). For example, the introduction of a trifluoroacetyl group is carried out by reacting with trifluoroacetic acid ethyl ester in a solvent such as methanol in the presence of a tertiary amine such as triethylamine at 0 ° C. to room temperature, preferably room temperature. In this reaction, R ³ may be introduced after protecting the free hydroxyl group with an appropriate protecting group, if necessary.

Reaction (G) The compound [11] is obtained by removing the amino protecting group R ³ of the compound represented by the general formula [10]. The method for removing the amino protecting group R ³ may vary depending on the kind of the protecting group, but a method known in peptide chemistry can be used, and mainly achieved by catalytic reduction or base treatment in consideration of the above reaction (F). To be done. For example, the removal of the trifluoroacetyl group is carried out in a solvent such as methanol using potassium carbonate, sodium carbonate, ammonia or the like as a base at 0 ° C. to reflux temperature, preferably room temperature.

Next, in the above, compound [7] and compound [10] are respectively compound [5] or compound

It is prepared by subjecting [9] to a condensation reaction with compound [6].
You may perform in steps as follows. That is, B is −
(CH ₂ ) _k −,

[Chemical 11] In other cases, as shown in the reaction (K), the compound represented by the general formula R ⁵ −
A compound represented by the general formula [17] is prepared by subjecting a compound represented by B-COOH (R ⁵ is a suitable amino protecting group) to a compound [5] in the same manner as in the case of the above reaction (C).
A compound represented by is obtained. Then, as shown in the reaction (L), the amino protecting group R ⁵ of the compound [17] is removed by a known method, and then the compound of the general formula W- (CH ₂ ) _n -A-H is used.
(When A is O, NH, CONH) or the general formula W-
Target compound [7] by reacting a compound represented by (CH ₂ ) _n —COOH (when A is a single bond)
Can be obtained. In the former case, for example, W-
(CH ₂ ) _n -A-H in a suitable solvent such as 1,4-dioxane or tetrahydrofuran in the presence of a tertiary amine such as triethylamine at -20 ° C to room temperature,
Reacts with phosgene, trichloromethyl, chloroformate, carbonyldiimidazole, etc., and then the compound [1
This can be achieved by reacting with the deprotected form of [7]. Further, when A is -NH- _{are, W- (CH 2) n -N} =
C = O in a suitable solvent such as dichloromethane, chloroform, N, N-dimethylformamide in the presence or absence of a tertiary amine such as triethylamine at -20 ° C.
To room temperature at room temperature to react with the deprotected form of compound [17]. If necessary, it may be carried out after protecting the free hydroxyl group of compound [17] with an appropriate protecting group. When A in the latter is a single bond, deprotection of the compound [17] using the method described in the above reaction (C), by the condensation reaction of the W- (CH _{2) n} -COOH To be achieved. On the other hand, B is-(CH
₂ ) _k- ,

[Chemical 12] In the case of, as shown in the reaction (M), the compound represented by the general formula R ⁶ —O
A compound represented by —CO—B—COOH (wherein R ⁶ is a suitable carboxyl protecting group) is subjected to a condensation reaction with compound [5] in the same manner as in the case of the above reaction (C) to give compound [1
8] is obtained. Then, as shown in the reaction (N), after removing the carboxyl protecting group R ⁶ of the present compound [18] by a conventional method, the reaction with the compound represented by the general formula W- (CH ₂ ) _n -AH is performed. Target compound by performing [7]
Can be obtained. Specifically W- _{(CH 2)} n -O
A conventional esterification reaction using H, W- (C
H ₂ ) _n- NH ₂ is used for the amide bond formation reaction described in the above reaction (C) or deprotection of compound [18] (R
⁶ ) The free carboxylic acid obtained in ⁶ ) is derivatized to acid chloride, active ester, etc. by a conventional method, and then W- (CH ₂ ) _n -N
This is achieved by carrying out a reaction with H ₂ . If necessary, it may be carried out after protecting the free hydroxyl group of the compound [18] with an appropriate protecting group. Also with respect to compounds Preparation of [10], B is - _{(CH 2)} k -,

[Chemical 13] Otherwise, the compound

From [9], a compound represented by the general formula [19] can be prepared in the same manner as in the above reaction (K), and then the reaction (L) can be carried out in the same manner as above to prepare the target compound [10]. it can. Also, B is-(CH
₂ ) _k- ,

[Chemical 14] In case of

Compound [10] can be prepared by subjecting [9] to the above reaction (M) to give compound [20], and then performing reaction (N) in the same manner as above.

The following general formula [1]

[Chemical 15] An optically active sulfinyl compound (X = SO)
Can be manufactured according to the steps shown below.

[0018]

[Chemical 16] Each of the above reaction steps will be further described. Here, the symbols A, B, W, R and n are the same as above, and the reactions (C), (D), (K), (L), (M) and (N) are as described above. Is.

Reaction (H) The compound represented by the general formula [12] is prepared by the method described in the literature (P. Pitchen et. Al., J. Am. Ch.
em. Soc. , 106 , 8188-8193 (198).
4); H. Zhao et. al. , Tetrah
edron, 43 , 5135-5144 (1987), etc.) to give a compound of general formula [13]
To obtain an optically active sulfoxide.
This reaction is carried out at 0 ° C. in the presence of titanium tetraisopropoxide, optically active diethyl tartrate and water in a solvent such as methylene chloride or 1,2-dichloroethane.
Hereafter, preferably at −40 ° C. to −20 ° C. t−
It is carried out using butyl hydroperoxide or cumene hydroperoxide.

Reaction (J) The optically active sulfoxide of the general formula [13] is treated with a base to give the corresponding carbanion, which is condensed with the ester or aldehyde of the general formula [14] or [15]. And the general formula [1] or [1
6] to obtain an optically active sulfinyl compound. This reaction is carried out, for example, by using an optically active sulfoxide [13] in an inert organic solvent such as THF and 1,4-dioxane at a reaction temperature of −78 ° C. to room temperature, preferably 0 ° C. or lower, n-butyllithium, The corresponding carbanion is formed using a base such as lithium diisopropylamide, and then ester [14] or aldehyde [15] is added to carry out the condensation reaction at -78 ° C to room temperature, preferably at -78 ° C to -20 ° C. It is achieved by Compounds used as synthetic raw materials, N-protected prolinal [2], HX-R [4], Q-COOH
_{^{[6], R 5 -B-COOH, W-}} (CH 2) n -A-
_{H, W- (CH 2) n} -A-COOH, CH 3 -S-R
Both [12] and an ester of proline are available as a substance known per se, or can be easily derived and synthesized from a known precursor by a known method. Further, when synthesizing the final target compound represented by the general formula [1], a protecting group may be introduced at an appropriate stage as necessary and the protecting group may be deleted at an appropriate step.
Isolation and purification of the thus obtained compound represented by the general formula [1] from the reaction mixture can be carried out in the field of synthetic organic chemistry. It can be carried out by using any commonly used means, and for example, it can be isolated and purified by a method such as column chromatography, solvent extraction, recrystallization and the like. Isolation and purification may be carried out for each reaction or after completion of some reactions. The above-mentioned series of compounds each have 1 to 3 asymmetric centers in the molecule, but in the present invention, the configuration of each asymmetric center is either R or S, or a mixture thereof. May be. Each optically active substance can be obtained by using an optically active compound as a starting material or by purifying the obtained diastereomeric mixture by a method such as column chromatography or recrystallization.

When the compound of the present invention is used as a medicine, it is usually administered systemically or locally, orally or parenterally. The dose varies depending on age, weight, symptoms, therapeutic effect, administration method, etc., but is usually once per adult per dose.
Orally administered once to several times a day in the range of 100 mg to 100 mg, or 0.2 mg to 20 m once per adult
It is administered parenterally once to several times a day in the range of g. The compound of the present invention is used in the form of solid composition for oral administration, liquid composition or injection for parenteral administration, suppository and the like. Solid compositions for oral administration include tablets, pills, capsules, powders, granules and the like. In such solid compositions one or more active substances are used in admixture with at least one inert diluent,
If necessary, an excipient, a binder, a lubricant, a disintegrant, a solubilizing agent, a stabilizer and the like may be contained. Tablets or pills
If necessary, it may be coated with a film of a gastric or enteric substance. Capsules include hard capsules and soft capsules. Liquid compositions for oral administration include solutions, emulsions, suspensions, syrups and elixirs. In such a liquid composition, a commonly used inert diluent is contained, and other than that,
It may also contain auxiliaries such as wetting agents and suspending agents, sweetening agents, flavoring agents, aromatic agents, and preservatives. Injections for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions and emulsions. In such an injection, one or more active substances are mixed with at least one inert aqueous diluent or an inert non-aqueous diluent, and if necessary, further preserved. Auxiliary agents such as agents, wetting agents, emulsifiers, dispersants, stabilizers and solubilizing agents may be contained. These are usually sterilized by filtration (bacterial retention filter, etc.), blending of a bactericide or gamma irradiation, or after these treatments, a solid composition is prepared by a method such as lyophilization, and sterilized immediately before use. Used by adding water or a sterile diluent for injection.

[0022]

EXAMPLES The present invention will be described in more detail below with reference to examples. The abbreviations used in the examples have the following meanings. DMF N, N-dimethylformamide DMSO dimethylsulfoxide THF tetrahydrofuran HOBt 1-hydroxybenzotriazole DCC N, N′-dicyclohexylcarbodiimide ¹ H NMR proton nuclear magnetic resonance spectrum CI-MS chemical ionization mass spectrometry spectrum EI-MS electron impact ionization mass spectrometry Spectrum

Example 1 (2S) -2- (4-chlorophenoxyacetyl) -1
-[N- (3-phenylpropionyl) -L-prolyl] pyrrolidine (Compound 1) a) (2S) -1- (tert-butoxycarbonyl)
-2- (1,2-epoxyethyl) pyrrolidine sodium hydride (NaH content 60%, 24.0 g)
DMSO (200 ml) was added to and the mixture was stirred at 70 ° C. for 1 hour. The reaction solution was returned to room temperature, THF (200 ml) was added, the temperature was adjusted to -5 ° C, and a DMSO solution (400 ml) of trimethylsulfonium iodide (122.5 g) was added dropwise. After stirring for 1 minute, TH of N- (tert-butoxycarbonyl) -L-prolinal (60.0 g) was added.
The F (200 ml) solution was added quickly and then stirred at 0 ° C. for 1 hour. The reaction solution was poured into ice water and extracted with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give the title compound as a crude product (54.9 g). This was used in the following reaction without purification.

B) (2S) -1- (tert-butoxycarbonyl) -2- [2- (4-chlorophenoxy)-
1-hydroxyethyl] pyrrolidine (2S) -1- (tert-butoxycarbonyl) -2
-(1,2-epoxyethyl) pyrrolidine (1.50
4-chlorophenol (1.90 g) and a 1 M sodium methoxide methanol solution (7.1 ml) were added to a methanol solution (5 ml) of g), and the mixture was refluxed at 70 ° C. for 19 hours. The reaction solution was poured into ice water and extracted with ethyl acetate. The organic layer was washed successively with 2N sodium hydroxide, saturated ammonium chloride and saturated brine, dried over anhydrous magnesium sulfate and concentrated to give the title compound as a crude product (1.95 g).

C) N- (3-phenylpropionyl)-
L-proline L-proline benzyl ester hydrochloride (20.0
Triethylamine (11.5 ml), 3-phenylpropionic acid (13.7 g) and DCC (18.8 g) were added to a methylene chloride solution (250 ml) of g) under ice cooling, and the mixture was stirred for 2 hours. After stirring at room temperature for 14 hours, the insoluble material was filtered off and the filtrate was concentrated. Dissolve the residue in ethyl acetate and add 1N
The extract was washed successively with hydrochloric acid, saturated aqueous sodium hydrogen carbonate and saturated brine, dried over anhydrous magnesium sulfate and concentrated. Silica gel column chromatography of the residue (eluent: hexane-ethyl acetate)
Purified in. Obtained N- (3-phenylpropionyl) -L-proline benzyl ester (26.1
g) 10% in 1% aqueous methanol solution (200 ml)
Pd-C (3.0 g) was added, and 2.
Stir for 5 hours. After removing the catalyst by filtration, the filtrate was concentrated to give the title compound (17.4 g).

D) (2S) -2- [2- (4-chlorophenoxy) -1-hydroxyethyl] -1- [N- (3
-Phenylpropionyl) -L-prolyl] pyrrolidine (2S) -1- (tert-butoxycarbonyl) -2
-[2- (4-chlorophenoxy) -1-hydroxyethyl] pyrrolidine (1.93 g) was added to 4N hydrochloric acid-1,4-.
It was dissolved in dioxane (20 ml) and stirred at room temperature for 45 minutes. The residue obtained by concentrating the reaction solution to dryness was added with methylene chloride (2
0 ml) and DMF (5 ml), N-methylmorpholine (0.7 ml), N- (3-phenylpropionyl) -L-proline (1.50 g) and HOBt (91).
5 mg) was added. The reaction solution was ice-cooled, water-soluble carbodiimide hydrochloride (1.30 g) was added, and the mixture was stirred for 2 hours. After stirring at room temperature for 14 hours, the reaction solution was concentrated. The residue was dissolved in ethyl acetate, washed successively with 1N hydrochloric acid, saturated aqueous sodium hydrogen carbonate and saturated brine, dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel column chromatography (eluent: chloroform-methanol) to give the title compound (1.
61 g) was obtained.

E) (2S) -2- (4-chlorophenynoxyacetyl) -1- [N- (3-phenylpropionyl) -L-prolyl] pyrrolidine (2S) -2- [2- (4- Chlorophenoxy) -1-
Hydroxyethyl] -1- [N- (3-phenylpropionyl) -L-prolyl] pyrrolidine (1.59 g) in DMSO (10 ml) was added to sulfur trioxide-pyridine complex (2.69 g) in DMSO (10 ml). Was added dropwise, and the mixture was stirred at room temperature for 1 hour. The reaction solution was poured into ice water and extracted with ethyl acetate. The organic layer was 1N HCl, saturated aqueous sodium hydrogen carbonate,
The extract was washed successively with saturated brine, dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel column chromatography (eluent: chloroform-methanol) to give the title compound (0.85 g).

Example (2S) -2- (4-hydroxyphenoxyacetyl)
-1- [N- (3-phenylpropionyl) -L-prolyl] pyrrolidine (Compound 2) a) (2S) -2- {2- [4- (benzyloxy) phenoxy] -1-hydroxyethyl} -1 -(Tert
-Butoxycarbonyl) pyrrolidine In the same manner as in Example 1, b), (2S) -1-
The title compound (1.72 g) was obtained from (tert-butoxycarbonyl) -2- (1,2-epoxyethyl) pyrrolidine (1.50 g) and 4- (benzyloxy) phenol (2.96 g).

B) (2S) -2- {2- [4- (benzyloxy) phenoxy] -1-hydroxyethyl} -1
-[N- (3-phenylpropionyl) L-prolyl]
Pyrrolidine In the same manner as in d) of Example 1, (2S) -2-
{2- [4- (benzyloxy) phenoxy] -1-hydroxyethyl} -1- (tert-butoxycarbonyl) pyrrolidine (1.71 g) and N- (3-phenylpropionyl) -L-proline (1.10 g) ) And a crude product of the title compound (1.82 g) was obtained.

C) (2S) -2- [4- (benzyloxy) phenoxyacetyl] -1- [N- (3-phenylpropionyl) -L-prolyl] pyrrolidine (2S) -2- {2- [4 Pyridine (0.3 ml) in DMSO solution (30 ml) of-(benzyloxy) phenoxy] -1-hydroxyethyl} -1- [N- (3-phenylpropionyl) -L-prolyl] pyrrolidine (1.81 g), Trifluoroacetic acid (0.15 ml) and DCC (1.2 g) were added, and the mixture was stirred at room temperature for 4 hours. The reaction solution was poured into ice water and extracted with ethyl acetate. 1 extract
It was washed successively with N hydrochloric acid, saturated aqueous sodium hydrogen carbonate and saturated brine, dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel column chromatography (eluent: chloroform-methanol) to give the title compound (1.45 g).

D) (2S) -2- (4-hydroxyphenoxyacetyl) -1- [N- (3-phenylpropionyl) -L-prolyl] pyrrolidine (2S) -2- [4- (benzyloxy) phenoxy Acetyl] -1- [N- (3-phenylpropionyl)-
Pd-Black (300 mg) was added to a mixed solution of L-prolyl] pyrrolidine (1.44 g) in menol (50 ml) -water (30 ml) -acetic acid (20 ml), and the mixture was stirred under a hydrogen atmosphere at room temperature for 4 hours. After concentrating the reaction mixture, the concentrate was poured into saturated aqueous sodium hydrogen carbonate and extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium hydrogen carbonate and saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The residue was purified by silica gel column chromatography (eluent: chloroform-methanol) to give the title compound (0.71 g).

Example 3 (2S) -1- [N- (4-chlorobenzylaminocarbonyl) -L-prolyl] -2- (4-chlorophenoxyacetyl) pyrrolidine (Compound 3) a) (2S) -1 -[N- (tert-Butoxycarbonyl) -L-prolyl] -2- [2- (4-chlorophenoxy) -1-hydroxyethyl] pyrrolidine In the same manner as in d) of Example 1, (2S) -1-
(Tert-Butoxycarbonyl) -2- [2- (4-
The title compound (2.13 g) was obtained from chlorophenoxy) -1-hydroxyethyl] pyrrolidine (2.80 g) and N- (tert-butoxycarbonyl) -L-proline (1.94 g).

B) (2S) -1- [N- (4-chlorobenzylaminocarbonyl) -L-prolyl] -2- [2
-(4-Chlorophenoxy) -1-hydroxyethyl]
Pyrrolidine trichloromethyl chloroformate (0.35 ml)
In THF solution (40 ml) under ice-cooling, 4-chlorobenzylamine (0.71 ml) and triethylamine (0.
A THF solution (20 ml) of 81 ml) was added dropwise, and the mixture was stirred at room temperature for 1.5 hours. (2S) -1- [N- (tert
-Butoxycarbonyl) -L-prolyl] -2- [2-
(4-Chlorophenoxy) -1-hydroxyethyl] pyrrolidine (2.13 g) was treated with 4N hydrochloric acid-1,4-dioxane (20 ml) to obtain (2S) -2- [2- (4.
-Chlorophenoxy) -1-hydroxyethyl] -1-
A methylene chloride suspension (20 ml) of (L-prolyl) pyrrolidine hydrochloride and triethylamine (0.81 ml) was added dropwise, and the mixture was stirred at room temperature for 1 hour. The reaction solution was poured into ice water and extracted with ethyl acetate. The organic layer was washed with 1N hydrochloric acid, saturated aqueous sodium hydrogen carbonate and saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The residue was purified by silica gel column chromatography (eluent: chloroform-methanol) to give the title compound (2.09 g).

C) (2S) -1- [N- (4-chlorobenzylaminocarbonyl) -L-prolyl] -2- (4
-Chlorophenoxyacetyl) pyrrolidine In the same manner as in the case of e) of Example 1, (2S)-[N-
(4-chlorobenzylaminocarbonyl) -L-prolyl] -2- [2- (4-chlorophenoxy) -1-hydroxyethyl] pyrrolidine (2.08 g) was added to sulfur trioxide-pyridine complex (3.27 g). Oxidation with used gave the title compound (1.20 g).

Example 4 (2S) -1- [N- (4-Fluorobenzylaminocarbonyl) -L-prolyl] -2- (4-fluorophenoxyacetyl) pyrrolidine (Compound 4) a) (2S) -1 -(Tert-butoxycarbonyl)
-2- [2- (4-Fluorophenoxy) -1-hydroxyethyl] pyrrolidine In the same manner as in Example 1, b), (2S) -1-
The title compound (1.9) was obtained from (tert-butoxycarbonyl) -2- (1,2-epoxyethyl) pyrrolidine (2.00 g) and 4-fluorophenol (2.21 g).
1 g) was obtained.

B) (2S) -1- [N- (tert-butoxycarbonyl) -L-prolyl] -2- [2- (4
-Fluorophenoxy) -1-hydroxyethyl] pyrrolidine In the same manner as in d) of Example 1, (2S) -1-
(Tert-Butoxycarbonyl) -2- [2- (4-
The title compound (1.85 g) was obtained from fluorophenoxy) -1-hydroxyethyl] pyrrolidine (1.90 g) and N- (tert-butoxycarbonyl) -L-proline (1.38 g).

C) (2S) -1- [N- (4-fluorobenzylaminocarbonyl) -L-prolyl] -2-
[2- (4-Fluorophenoxy) -1-hydroxyethyl] pyrrolidine In the same manner as in b) of Example 3, 4-fluorobenzylamine (0.54 ml) and trichloromethyl chloroformate (0.28 ml). And (2S) -1-
The title compound (1.38 g) was obtained from [N- (tert-butoxycarbonyl) -L-prolyl] -2- [2- (4-fluorophenoxy) -1-hydroxyethyl] pyrrolidine (1.85 g). ..

D) (2S) -1- [N- (4-fluorobenzylaminocarbonyl) -L-prolyl] -2-
(4-Fluorophenoxyacetyl) pyrrolidine In the same manner as in (e) of Example 1, (2S) -1-
[N- (4-fluorobenzylaminocarbonyl) -L
-Prolyl] -2- [2- (4-fluorophenoxy)
-1-Hydroxyethyl] pyrrolidine (1.36 g) was oxidized with sulfur trioxide-pyridine complex (1.83 g) to obtain the title compound (1.15 g).

Example 5 (2S) -1- [N- (benzylaminocanibonyl)-
L-valyl] -2- (phenoxyacetyl) pyrrolidine (Compound 5) a) (2S) -1- (tert-butoxycarbonyl)
-2- (1-Hydroxy-2-phenoxyethyl) pyrrolidine In the same manner as in Example 1, b), (2S) -1-
The title compound (2.20 g) was obtained from (tert-butoxycarbonyl) -2- (1,2-epoxyethyl) pyrrolidine (3.00 g) and phenol (3.00 g).

B) (2S) -1- [N- (tert-butoxycarbonyl) -L-valyl] -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine Same as in d) of Example 1. Then, (2S) -1-
(Tert-Butoxycarbonyl) -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (2.00
The title compound (1.92 g) was obtained from g) and N- (tert-butoxycarbonyl) -L-valine (1.55 g).

C) (2S) -1- [N- (benzylaminocarbonyl) -L-valyl] -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (2S) -1- [N- (tert- Butoxycarbonyl) -L-valyl] -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (1.90 g) was dissolved in 4N hydrochloric acid 1,4-dioxane (12 ml), and the mixture was stirred at room temperature for 40%.
Stir for minutes. The reaction solution was concentrated to dryness. A solution of the residue in methylene chloride (30 ml) was added to triethylamine (0.5 ml) and benzyl isocyanate (0.58) under ice cooling.
ml) was sequentially added dropwise. After stirring for 2 hours, the reaction solution was poured into ice water and extracted with ethyl acetate. The organic layer was washed successively with 1N hydrochloric acid, saturated aqueous sodium hydrogen carbonate and saturated brine, dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel column chromatography (eluent: chloroform-methanol) to give the title compound (1.55 g).

D) (2S) -1- [N- (benzylaminocarbonyl) -L-valyl] -2- (phenoxyacetyl) pyrrolidine (2S) -1 in the same manner as in e) of Example 1. −
[N- (benzylaminocarbonyl) -L-valyl]-
2- (1-Hydroxy-2-phenoxyethyl) pyrrolidine (1.54 g) was oxidized with sulfur trioxide-pyridine complex (2.23 g) to give the title compound (819m
g) was obtained.

Example 6 (2S) -1- [N- (4-chlorobenzylaminocarbonyl) -L-prolyl] -2- (phenoxyacetyl) pyrrolidine (Compound 6) a) (2S) -1- [N -(Tert-Butoxycarbonyl) -L-prolyl] -2- (1-hydroxy-2-
Phenoxyethyl) pyrrolidine In the same manner as in d) of Example 1, (2S) -1-
(Tert-Butoxycarbonyl) -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (1.93
The title compound (2.10 g) was obtained from g) and N- (tert-butoxycarbonyl) -L-proline (1.36 g).

B) (2S) -1- [N- (4-chlorobenzylaminocarbonyl) -L-prolyl] -2- (1
-Hydroxy-2-phenoxyethyl) pyrrolidine In the same manner as in b) of Example 3, 4-chlorobenzylamine (0.71 g), trichloromethyl chloroformate (0.3 ml) and (2S) -1-. [N-,
(Tert-Butoxycarbonyl) -L-prolyl]-
The title compound (1.13 g) was obtained from 2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (2.00 g).

C) (2S) -1- [N- (4-chlorobenzylaminocarbonyl) -L-prolyl] -2- (phenoxyacetyl) pyrrolidine In the same manner as in e) of Example 1, (2S) ) -1-
[N- (4-chlorobenzylaminocarbonyl) -L-
Prolyl] -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (1.10 g) was oxidized using sulfur trioxide-pyridine complex (1.85 g) to obtain the title compound (0.75 g).

Example 7 (2S) -1- (N-octanoyl-L-prolyl)-
2- (phenoxyacetyl) pyrrolidine (Compound 7) a) N-octanoyl-L-proline benzyl ester L-proline benzyl ester (7.37 g) in a methylene chloride solution (80 ml) was cooled with ice and triethylamine (4.3 ml). ), Octanoic acid (4.8 ml) and D
CC (6.30 g) was added successively and the mixture was stirred for 1 hour. After stirring at room temperature for 15 hours, the insoluble material was filtered off. The filtrate was diluted with chloroform, washed successively with 1N hydrochloric acid, saturated aqueous sodium hydrogen carbonate and saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The residue was purified by silica gel column chromatography (eluent: n-hexane-ethyl acetate) to give the title compound (8.0
9 g) was obtained.

B) N-octanoyl-L-proline N-octanoyl-L-proline benzyl ester (7.04 g) in 1% aqueous methanol solution (70 ml).
10% Pd-C (0.71 g) was added to and the mixture was stirred at a high temperature under a hydrogen atmosphere for 1.5 hours. After removing the catalyst by filtration, the filtrate was concentrated to give the title compound (5.08 g).

C) (2S) -2- (1-hydroxy m2)
-Phenoxyethyl) -1- (N-octanoyl-L-
Prolyl) pyrrolidine In the same manner as in d) of Example 1, (2S) -1-
(Tert-Butoxycarbonyl) -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (1.50
g) and N-octanoyl-L-proline (1.18 g)
The title compound (1.16 g) was obtained.

D) (2S) -1- (N-octanoyl-
L-prolyl) -2- (phenoxyacetyl) pyrrolidine In the same manner as in e) of Example 1, (2S) -2-
(1-hydroxy-2-phenoxyethyl) -1- (N
-Octanoyl-L-prolyl) pyrrolidine (1.11
g) was oxidized with sulfur trioxide-pyridine complex (2.70 g) to give the title compound (0.78 g).

Example 8 (2S) -1- [N- (benzylaminocarbonyl)-
L-prolyl] -2- (benzyloxyacetyl) pyrrolidine (Compound 8) a) (2S) -2- (2-benzyloxy-1-hydroxyethyl) pyrrolidine benzyl alcohol (2.0 ml) in DMF solution (10
ml) with sodium hydride (60% dispersi
on in oil, 378 mg) was added, and the mixture was stirred at room temperature for 1 hour. A DMF solution (10 ml) of (2S) -1- (tert-butoxycarbonyl) -2- (1,2-epoxyethyl) pyrrolidine (2.00 g) was added dropwise and stirred for 4 hours. Pour the reaction solution into saturated ammonium chloride,
It was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The compound (640 mg) obtained by purifying the residue by silica gel column chromatography (eluent: hexane-ethyl acetate) was dissolved in 1,4-dioxane-water (1: 1, 30 ml),
Add barium hydroxide octahydrate (800 mg) to 120
Reflux at 4 ° C. for 4 hours. After removing the insoluble matter by filtration, the pH of the filtrate is adjusted to 1
It was adjusted to 5 to 6 with M phosphoric acid and washed with ether. Then, the aqueous layer was adjusted to pH ~ 8 with 1N sodium hydroxide and extracted with chloroform. The chloroform extract was dried over anhydrous magnesium sulfate and concentrated to give crude crystals of the title compound (2
97 mg) was obtained.

B) (2S) -1- [N- (tert-butoxycarbonyl) -L-prolyl] -2- (2-benzyloxy-1-hydroxyethyl) pyrrolidine As in Example 1 d). Then, (2S) -2-
(2-Benzyloxy-1-hydroxyethyl) pyrrolidine (297 mg) and N- (tert-butoxycarbonyl) -L-proline (318 mg) were dehydrated and condensed to obtain the title compound (274 mg).

C) (2S) -1- [N- (benzylaminocarbonyl) -L-prolyl] -2- (2-benzyloxy-1-hydroxyethyl) pyrrolidine As in c) of Example 5. Then, (2S) -1-
The title compound (243 mg) was obtained from [N- (tert-butoxycarbonyl) -L-prolyl] -2- (2-benzyloxy-1-hydroxyethyl) pyrrolidine (274 mg) and benzyl isocyanate (89 μl).

D) (2S) -1- [N- (benzylaminocarbonyl) -L-prolyl] -2- (benzyloxyacetyl) pyrrolidine In the same manner as in Example 1, e), (2S)- 1-
[N- (benzylaminocarbonyl) -L-prolyl]
-2- (2-benzyloxy-1-hydroxyethyl)
Pyrrolidine (233 mg) was oxidized with sulfur trioxide-pyridine complex (330 mg) to give the title compound (106
mg) was obtained.

Example 9 (2S) -1- [N- (benzylaminocarbonyl)-
L-prolyl] -2-{[(3,4-methylenedioxy) phenoxy] acetyl} pyrrolidine (Compound 9) a) (2S) -1- (tert-butoxycarbonyl)
-2- {1-hydroxy-2-[(3,4-methylenedioxy) phenoxy] ethyl} pyrrolidine In the same manner as in Example 1, b), (2S) -1-
(Tert-Butoxycarbonyl) -2- (1,2-epoxyethyl) pyrrolidine (2.00 g) and (3,4-
The title compound (2.41 g) was obtained from methylenedioxy) phenol (2.72 g).

B) (2S) -1- [N- (tert-butoxycarbonyl) -L-prolyl] -2- {1-hydroxy-2-[(3,4-methylenedioxy) phenoxyethyl} pyrrolidine Implementation Similarly to the case of d) of Example 1, (2S) -1-
(Tert-Butoxycarbonyl) -2- {1-hydroxy-2-[(3,4-methylenedioxy) foxy]
Ethyl} pyrrolidine (2.38 g) and N- (tert-
The title compound (2.07 g) was obtained from butoxycarbonyl) -L-proline (1.60 g).

C) (2S) -1- [N- (benzylaminocarbonyl) -L-prolyl] -2- {1-hydroxy-2-[(3,4-methylenedioxy) phenoxy]
Ethyl} pyrrolidine In the same manner as in the case of c) of Example 5, (2S) -1-
[N- (tert-butoxycarbonyl) -L-prolyl] -2- {1-hydroxy-2-[(3,4-methylenedioxy) phenoxy] ethyl} pyrrolidine (2.0
The title compound (2.06 g) was obtained from 7 g) and benzyl isocyanate (0.63 ml).

D) (2S) -1- [N- (benzylaminocarbonyl) -L-prolyl] -2-{[(3,4-
Methylenedioxy) phenoxy] acetyl} pyrrolidine In the same manner as in the case of e) in Example 1, (2S) -1-
[N- (benzylaminocarbonyl) -L-prolyl]
-2- {1-hydroxy-2-[(3,4-methylenedioxy) phenoxy] ethyl} pyrrolidine (2.05
g) was oxidized with sulfur trioxide-pyridine complex (2.71 g) to give the title compound (1.12 g).

Example 10 (2S) -1- [2- (benzylaminocarbonyl) benzoyl] -2- (phenoxyacetyl) pyrrolidine (Compound 10) a) 2- (Benzylaminocarbonyl) benzoic acid Phthalic anhydride (5 0.000 g) in methylene chloride (50
benzylamine (3.70 g) was added to (ml) and the mixture was stirred at room temperature for 15 minutes. The precipitated product was collected by filtration to give the title compound (9.09 g).

B) (2S) -1- [2- (benzylaminocarbonyl) benzoyl] -2- (1-hydroxy-
2-phenoxyethyl) pyrrolidine In the same manner as in d) of Example 1, (2S) -1-
(Tert-Butoxycarbonyl) -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (1.27
The title compound (1.10 g) was obtained from g) and 2- (benzylaminocarbonyl) benzoic acid (0.76 g).

C) (2S) -1- [2- (benzylaminocarbonyl) benzoyl] -2- (phenoxyacetyl) pyrrolidine (2S) -1-in the same manner as in Example 1, e).
[2- (benzylaminocarbonyl) benzoyl] -2
-(1-Hydroxy-2-phenoxyethyl) pyrrolidine (1.09 g) was mixed with sulfur trioxide-pyridine complex (1.
Oxidation with 56 g) gave the title compound (0.79 g).

Example 1 (2S) -2- (phenoxyacetyl) -1- {2-
[(Phenylacetyl) amino] benzoyl} pyrrolidine (Compound 11) a) 2-[(Phenylacetyl) amino] benzoic acid 2-Aminobenzoic acid (3.00 g) in methylene chloride suspension (90 ml) in triethylamine (3 .3 ml)
And phenylacetic acid chloride (2.90 ml) were added dropwise,
Stir at room temperature for 4 hours. The reaction solution was poured into water, adjusted to pH 11 with 1N sodium hydroxide, and then extracted. The aqueous layer was adjusted to pH 3 with concentrated hydrochloric acid and then extracted with chloroform. The chloroform layer was washed with saturated brine, dried over anhydrous magnesium sulfate and concentrated. The residue was crystallized from hexane to give the title compound (1.56 g).

B) (2S) -2- (1-hydroxy-2)
-Phenoxyethyl) -1- {2-[(phenylacetyl) amino] benzoyl} pyrrolidine In the same manner as in d) of Example 1, (2S) -1-
(Tert-Butoxycarbonyl) -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (1.89
g), 2-[(phenylacetyl) amino] benzoic acid (1.30 g) and the title compound (315 mg) were obtained.

C) (2S) -2- (phenoxyacetyl) -1- {2-[(phenylacetyl) amino] benzoyl} pyrrolidine In the same manner as in Example 1, e), (2S) -2-
(1-Hydroxy-2-phenoxyethyl) -1- {2
-[(Phenylacetyl) amino] benzoyl} pyrrolidine (315 mg) was mixed with sulfur trioxide-pyridine complex (4
50 mg) to give the title compound (164 mg)
Got The physicochemical properties of these compounds 1 to 11 are shown in Table 1.
Through Table 3.

[0064]

[Table 1]

[0065]

[Table 2]

[0066]

[Table 3] Further, it is needless to say that the present invention is not limited to these examples. For example, the compounds 12 to 2 shown in Table 4 can be used.
7 also belongs to the present invention.

[0067]

[Table 4]

Next, a test was conducted on the prolyl endopeptidase inhibitory activity and the inhibitory activity against various proteolytic enzymes of the proline derivative represented by the general formula [1] according to the present invention in an in vitro system. Test Example 1 Prolyl endopeptidase inhibitory activity 0.1 M potassium-sodium phosphate buffer (pH 7.
0) 2675 μl, 100 μl of 0.1 M potassium-sodium phosphate buffer solution (pH 7.0) of the compound of the present invention
And a solution of prolyl endopeptidase extracted from rat brain in 25 mM sodium phosphate buffer (123 units / l, pH 6.8, 1 mM dithiothreitol and 0.
Contains 5 mM EDTA. J. Neurochem. ，
35 , 527 (1980)) 10
0 μl of the mixture was preincubated at 30 ° C. for 30 minutes. To this, 125 μl of a 0.1 M potassium phosphate-sodium phosphate buffer solution (pH 7.0) of 0.2 mM 7- (N-succinyl-glycyl-prolyl) -4-methylcoumarinamide (manufactured by Peptide Institute, Inc.) In addition, the mixture was incubated at 30 ° C for 1 hour. The reaction solution was immersed in ice (0 ° C.) to stop the reaction, and 10 minutes later, the fluorescence intensity (a at excitation wavelength 370 nm and fluorescence wavelength 440 nm) (a
₁ ) was measured. At the same time, in the above system, an experiment using 25 mM sodium phosphate buffer (containing pH 6.8, 1 mM dithiothreitol and 0.5 mM EDTA) instead of the prolyl endopeptidase solution, and instead of the compound solution of the present invention Experiments were carried out using a 0.1 M potassium-sodium phosphate buffer (pH 7.0), and the fluorescence intensities a ₂ and a ₃ were measured (protein nucleic acid enzyme, 2
9 , 127 (1984)). The prolyl endopeptidase inhibition rate was calculated by the formula 1, and the concentration required for 50% inhibition (IC ₅₀ ) was determined using a semilogarithmic graph. The test results are shown in Table 5.

[0069]

[Equation 1]

[0070]

[Table 5] As is clear from the results of this test, it was confirmed that the compound of the present invention has an excellent inhibitory activity against prolyl endopeptidase.

Test Example 2 Inhibitory activity against various proteolytic enzymes The compounds of the present invention were tested for specificity of the inhibitory activity against various proteolytic enzymes at a predetermined concentration. It was found to specifically inhibit peptidases.

[0072]

[Table 6] The method for measuring the inhibitory activity of various proteolytic enzymes excluding prolyl endopeptidase and the method for calculating the inhibition rate are as follows.

Measurement of Trypsin Inhibitory Activity In this test, 50 mM Tris-HCl buffer (pH 8.0) was used as the measurement buffer. Same buffer 8
0.02 μM of 50 μl, 50 μl of the same buffer solution of the compound of the present invention and trypsin (derived from bovine pancreas, manufactured by Sigma)
To a mixed solution of 50 μl of the same buffer solution, 50 μl of 200 μM same-buffer solution of 7- (prolyl-phenylalanyl-arginyl) -4-methylcoumarinamide (manufactured by Peptide Institute Co., Ltd.) was added, and the mixture was added at 30 ° C. at 1 ° C. Incubated for hours. The reaction solution was immersed in ice (0 ° C.) to stop the reaction, and 1 hour later, the fluorescence intensity (b ₁ ) at an excitation wavelength of 370 nm and a fluorescence wavelength of 440 nm was measured. At the same time, in the above system, an experiment using the same buffer solution instead of the trypsin solution, and an experiment using the same buffer solution instead of the compound solution of the present invention,
Similarly, the fluorescence intensities (b ₂ ) and (b ₃ ) were measured.

Measurement of chymotrypsin inhibitory activity 50 mM Tris-HCl buffer (pH
8.0) as an enzyme solution and a 0.2 μM buffer solution of chymotrypsin (derived from bovine pancreas, manufactured by Sigma) as a substrate solution, 7- (N-succinyl-leucyl-leucyl-).
Using a 200 μM buffer solution of valyl-tyrosyl) -4-methylcoumarinamide (manufactured by Peptide Research Co., Ltd.) in the same manner as above, the fluorescence intensities c ₁ and c were obtained.
₂ and c ₃ were measured.

Measurement of Leucine Aminopeptidase Inhibitory Activity 50 mM Tris-HCl buffer (pH
8.0) as a enzyme solution, a 0.2 μM buffer solution of leucine aminopeptidase (derived from pig kidney, manufactured by Sigma) as a substrate solution, and as a substrate solution, 7-leucyl-4-methylcoumarinamide (Peptide Research Institute, Inc.) (Manufactured by Mitsui Chemicals Co., Ltd.), and the fluorescence intensities d ₁ , d ₂ and d ₃ were measured in exactly the same manner as above.

Measurement of Elastase Inhibitory Activity 1 mM Tris-HCl buffer (pH
8.5) as an enzyme solution and elastase (derived from porcine pancreas,
Sigma's 0.2 μM same buffer solution is used as a substrate solution of 7- (N-succinyl-alanyl-prolyl-alanyl) -4-methylcoumarinamide (manufactured by Peptide Institute Co., Ltd.) 200 μM same buffer Using the solution, the fluorescence intensities e ₁ , e ₂ and e ₃ were measured in exactly the same manner as above.

Measurement of Cathepsin B Inhibitory Activity 100 mM sodium phosphate buffer solution (pH 6.0; containing 1.33 mM EDTA · Na ₂ ) was used as a measurement buffer, and cathepsin B (derived from bovine spleen as an enzyme solution,
Sigma's 0.02 μM buffer solution was used as a substrate solution of 7- (N-benzyloxycarbonyl-phenylalanyl-arginyl) -4-methylcoumarinamide (manufactured by Peptide Research Co., Ltd.) at 200 μM. Using a buffer solution, the fluorescence intensities f ₁ and f were measured in the same manner as above.
₂ and f ₃ were measured. Using the fluorescence intensities x ₁ , x ₂ and x ₃ (x represents b, c, d, e and f) thus measured, the inhibition rate against various proteolytic enzymes was calculated by
Calculated by

[0078]

[Equation 2]

[0079]

INDUSTRIAL APPLICABILITY The novel proline derivative represented by the above general formula [1] according to the present invention has very strong inhibitory activity against prolyl endopeptidase, but it has trypsin, chymotrypsin, leucine aminopeptidase, elastase, It has been observed that it has no effect on proteases such as cathepsin B, and thus hormones in the brain containing proline residues, neurotransmitters such as TR
It is considered to be a compound that specifically suppresses the decomposition and inactivation of H, substance P, neurotensin, vasopressin and the like. Therefore, the compound of the present invention can be expected to make an effective contribution to the improvement of symptoms of various diseases mediated by hormones and neurotransmitters, and as an anti-dementia drug or anti-amnestic agent that directly acts on the core symptoms of dementia, Alzheimer's disease. It can be used for the prevention and / or treatment of dementia and amnesia, including.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification number Reference number within the agency FI Technical indication location C12N 9/99 (72) Inventor Kunio Iwata 23 Furuki, Hadano City, Kanagawa Japan Tobacco Inc. Safety (72) Inventor Ituro Uchida 6-2 Umegaoka, Midori-ku, Yokohama-shi, Kanagawa Japan Tobacco Inc. Pharmaceutical Research Institute

Claims (2)

[Claims] 1. A general formula [1]: [In the formula, A is -O-, -NH-, -CONH- or a single bond, and B is - _{(CH 2) K} - or -NHCHR ¹ - a, (however, k
Is an integer of 1 to 3, R ¹ is a hydrogen atom or a lower alkyl group), and W is Or CH ₃ — (wherein R ² represents a hydrogen atom, a halogen atom or a lower alkoxy group), X is —S—, —S.
_{O -, - SO 2 -,} - O- or -NH- and, R represents ## STR4 ## Or a lower alkyl group (wherein 1 is an integer of 0 to 3,
Y and Z are the same or different and each is a hydrogen atom, a halogen atom,
It means a lower alkyl group which may be substituted with a fluorine atom, an amino group, a nitro group, a hydroxyl group or a lower alkoxy group.
Further, Y and Z may combine with each other to form a saturated or unsaturated 5- and 6-membered ring, and n represents an integer of 1 to 6). 2. (2S) -2- (4-chlorophenoxyacetyl) -1- [N- (3-phenylpropionyl)-
L-prolyl] pyrrolidine, (2S) -2- (4-hydroxyphenoxyacetyl) -1- [N- (3-phenylpropionyl) -L-prolyl] pyrrolidine, (2
S) -1- [N- (4-chlorobenzylaminocarbonyl) -L-prolyl] -2- (4-chlorophenoxyacetyl) pyrrolidine, (2S) -1- [N- (4-fluorobenzylaminocarbonyl) -L-prolyl] -2
-(4-fluorophenoxyacetyl) pyrrolidine,
(2S) -1- [N- (benzylaminocarbonyl)-
L-valyl] -2- (phenoxyacetyl) pyrrolidine, (2S) -1- [N- (4-chlorobenzylaminocarbonyl) -L-prolyl] -2- (phenoxyacetyl) pyrrolidine, (2S) -1- (N-Octanoyl-L-prolyl) -2- (phenoxyacetyl) pyrrolidine, (2S) -1- [N- (benzylaminocarbonyl) -L-prolyl] -2- (benzyloxyacetyl) pyrrolidine, (2S) -1- [N- (benzylaminocarbonyl) -L-prolyl] -2-{[(3,4-
Methylenedioxy) phenoxy] acetyl} pyrrolidine, (2S) -1- [2- (benzylaminocarbonyl) benzoyl] -2- (phenoxyacetyl) pyrrolidine and (2S) -2- (phenoxyacetyl) -1-
The novel proline derivative and a salt thereof according to claim 1, which are selected from {2-[(phenylacetyl) amino] benzoyl} pyrrolidine.