JPH05186498A - Proline derivative - Google Patents

Abstract

(57) [Summary] (Modified) [Structure] General formula [A is —O—, —CR ¹ — or —NR ² — (R ¹ is H or a heterocycle, R ² is a lower alkoxycarbonyl lower alkyl group); W is substituted with a halogen, a lower alkyl group or a lower alkoxy group. Optionally a phenyl group, an adamantyl group, a lower alkyl group or a heterocycle; U is -O-, -S
-, - NH- or -CHR ⁵ - ^{(R 5} is H or a lower alkoxycarbonyl group); X is -S -, - SO -, - SO
_2- , -O- or -NH-; D is a benzene ring or a heterocycle; Y and Z are the same or different and are 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]. [Effect] This proline derivative has a strong inhibitory activity specifically on prolyl endopeptidase, suppresses the degradation and inactivation of TRH, substance P, neurotensin, pasopressin, etc., and anti-dementia including Alzheimer's disease. It can be used as a drug and amnestic drug for preventive treatment.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a proline compound having a 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 this enzyme can be obtained, these compounds suppress the decomposition and inactivation of vasopressin and the like, and directly act on the core symptoms of dementia. As a drug, it is expected to be applicable to 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 JP-A-8467, JP-A-84-42475, 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 4] [In the formula, A is —O—, —CHR ¹ — or —NR ² — (wherein R ¹ is a hydrogen atom or a heterocycle, and R ² is a lower alkoxycarbonyl lower alkyl group); B
Is

[Chemical 5] Or —NR ³ —CHR ⁴ — (wherein R ³ and R ^{4 are}
Are the same or different and are a hydrogen atom or a lower alkyl group); W is a phenyl group which may be substituted with a halogen atom, a lower alkyl group or a lower alkoxy group, an adamantyl group, a lower alkyl group or a heterocycle; U
Is —O—, —S—, —NH— or —CHR ⁵ — (wherein R ⁵ is a hydrogen atom or a lower alkoxycarbonyl group); X is —S—, —SO—, —SO ₂ —. , -O
-Or -NH-; R is

[Chemical 6] (Where D is a benzene ring or a heterocycle, and Y and Z
Are the same or different and are a lower alkyl group, an amino group, a nitro group, a hydroxyl group or a lower alkoxy group which may be substituted with a hydrogen atom, a halogen atom or a fluorine atom). However, when U is —CH ₂ —, at least W is an adamantyl group or a heterocycle, or A is —CHR ¹ — or —NR ² — (wherein R ¹ is a heterocycle, R is ² is a lower alkoxycarbonyl lower alkyl group), B is a thiazolidine, or R means any of a heterocycle]]. Another object is to provide a pharmaceutical composition containing the above proline derivative [1] as an active ingredient and useful as a prolyl endopeptidase inhibitor.

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. "Lower alkoxycarbonyl group"
Is an alkoxycarbonyl group having 2 to 6 carbon atoms, specifically, methoxycarbonyl group, ethoxycarbonyl group, propoxycarbonyl group, isopropoxycarbonyl group, butoxycarbonyl group, sec-butoxycarbonyl group, tert-butoxycarbonyl group. Group etc. The “lower alkoxycarbonyl lower alkyl group” is an alkyl group having 1 to 5 carbon atoms substituted with a lower alkoxycarbonyl group as described above, and specifically, a methoxycarbonylmethyl group or a 2- (methoxycarbonyl) ethyl group. , 3- (methoxycarbonyl) propyl group, ethoxycarbonylmethyl group, 2- (ethoxycarbonyl) ethyl group, 3- (ethoxycarbonyl) propyl group, propoxycarbonylmethyl group, 2- (propoxycarbonyl) ethyl group, 3- ( Propoxycarbonyl) propyl group and the like. "Halogen atom" means chlorine,
Bromine, fluorine and iodine. The "phenyl group which may be substituted with a halogen atom, a lower alkyl group or a lower alkoxy group" is a phenyl group which may be substituted with 1 to 2 halogen atoms, a lower alkyl group or an alkoxy group as described above. , Specifically, 2-chlorophenyl group, 3-chlorophenyl group, 4-chlorophenyl group, 2-fluorophenyl group, 3-fluorophenyl group, 4-fluorophenyl group, 2-bromophenyl group,
3-bromophenyl group, 4-bromophenyl group, 2-iodophenyl group, 3-iodophenyl group, 4-iodophenyl group, 2-methylphenyl group, 3-methylphenyl group, 4-methylphenyl group, 2- Ethylphenyl group, 3
-Ethylphenyl group, 4-ethylphenyl group, 2-propylphenyl group, 3-propylphenyl group, 4-propylphenyl group, 2-isopropylphenyl group, 3-isopropylphenyl group, 4-isopropylphenyl group, 2
-Butylphenyl group, 3-butylphenyl group, 4-butylphenyl group, 2-methoxyphenyl group, 3-methoxyphenyl group, 4-methoxyphenyl group, 2-ethoxyphenyl group, 3-ethoxyphenyl group, 4-ethoxy Phenyl group, 3,4-dichlorophenyl group, 3,4-difluorophenyl group, 3,4-dibromophenyl group, 3,4
-Diiodophenyl group, 3,4-dimethylphenyl group,
3,4-diethylphenyl group, 3,4-dipropylphenyl group, 3,4-diisopropylphenyl group, 3,4-
Dibutylphenyl group, 3,4-dimethoxyphenyl group,
3,4-dimethoxyphenyl group, 3,4-dipropoxyphenyl group, 3,4-diisopropoxyphenyl group,
3,4-dibutoxyphenyl group and the like. "Heterocycle"
Means a 4- to 7-membered ring containing one or more heteroatoms, and specifically, pyrrole, furan, thiophene, pyrazole, isoxazole, imidazole, oxazole, thiazole, pyridine, pyrimidine, pyrazine, azetidine, pyrrolidine. , Tetrahydrofuran, piperidine, piperazine, morpholine, homopiperidine and the like. The proline compound 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 7]

[0009]

[Chemical 8] Each of the above reaction steps will be further described. In addition,
The symbols A, B, W, U, X and R have the same meanings as described above.

Reaction (A) The compound represented by the general formula [2] is reacted with sulfaylide derived from trimethylsulfonium iodide or trimethylsulfoxonium iodide in the presence of a strong base to give the compound represented by the general formula [3]. To obtain the epoxide represented by. In this reaction, for example, n-butyllithium or sodium hydride-dimethyl sulfoxide is used in an inert solvent such as tetrahydrofuran, 1,4-dioxane, or hexane to give a sulfaylide from the above sulfonium salt, and then the compound [ It is achieved by reacting with 2]. The reaction temperature is -70 ° C to reflux temperature, preferably -10 ° C to room temperature. Where R
⁶ is an amino-protecting group such as benzyloxycarbonyl group, tert-butoxycarbonyl group, 9-fluorenylmethyloxycarbonyl group, etc., and may be any amino-protecting group as long as it does not interfere with the reaction. The compound [3] can also be obtained by subjecting the compound [2] to a Wittig reaction to give an olefin and then epoxidizing it with a peracid. Specifically, for example, in an inert solvent such as tetrahydrofuran or diethyl ether, n
-The olefin is obtained by generating the corresponding ylide from methyltriphenylphosphonium halide using -butyllithium or the like, and then reacting it with the compound [2]. The reaction temperature is appropriately from -70 ° C to the reflux temperature.
This olefin is converted into methylene chloride, benzene, hexane,
The objective is to carry out the epoxidation reaction in an organic peracid such as m-chloroperbenzoic acid or hydrogen peroxide solution in a solvent such as methanol at a temperature of -20 ° C to reflux temperature, preferably 0 ° C to room temperature. The compound [3] can be obtained.

Reaction (B) The compound represented by the general formula [3] is 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, anion HO-R with a solvent sodium hydride such as N- dimethylformamide ^{▲ - ▼} O After setting -R [3]
React with. Reaction with H ₂ N-R X is NH, methanol, it is carried out in a solvent such as 1,4-dioxane. The reaction temperature is room temperature to reflux temperature in each case.

Reaction (C) The amino protecting group R ⁶ of the intermediate represented by the general formula [5] is removed according to a known method, and this is condensed with the compound represented by the formula [6] to give the compound [7]. ] Is obtained. Similarly, the general formula

The compound [10] can be obtained from the compound represented by [9]. When the amino-protecting group R ⁶ is, for example, a tert-butoxycarbonyl group (Boc group), its removal is carried out by the formula [5] or

The intermediate represented by [9] is converted to hydrobromic acid / acetic acid, hydrochloric acid / 1, by a known method.
It can be removed by acid treatment using 4-dioxane, formic acid, hydrochloric acid / acetic acid, trifluoroacetic acid or the like at -30 ° C to 70 ° C, preferably 0 ° C to 30 ° C. Next, the deprotected product thus obtained is converted into a compound [6] by a conventional method.
Amino acid derivative [7] or [1] by condensation reaction with
0] is obtained. Here, Q is W-UA-CO-B-
Means a group. For this peptide formation reaction, a method known per se can be adopted. N,
N'-dicyclohexylcarbodiimide (DCC), water-soluble carbodiimide hydrochloride (1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC)
・ HCl)) as a condensing agent, active ester method,
A mixed acid anhydride method and the like can be mentioned. The reaction is carried out in an inert solvent,
It is performed at 0 ° C to under heating. As a suitable solvent, 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 without isolation, and the above deprotected product and an inert solvent at 0 ° C.
The reaction is carried out at 40 to 40 ° C. to form a peptide bond. The mixed acid anhydride method is carried out in the presence of a tertiary amine (eg, pyridine, triethylamine) in an inert solvent,
The above compound [6] is reacted with an acid halide (eg, pivaloyl chloride, tosyl chloride, oxalyl chloride) or an acid derivative (eg, ethyl chloroformate, isobutyl chloroformate) at 0 ° C to 40 ° C to form a mixed acid. It is an anhydride, and this mixed acid anhydride is reacted with the above-mentioned deprotected form 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 (H
ONB)) is added or not added, and a desired peptide bond is formed by reacting [6] with the above deprotected form using DCC, EDC.HCl or the like as a condensing agent.

Reaction (D) By oxidizing the alcohol compound represented by the general formula [7] obtained in the above reaction (C) with an appropriate oxidizing agent,
The final target compound [1] is obtained. Similarly, the final target compound [1] can be obtained by oxidizing the compound [8] or the compound [11] described below. This reaction is carried out, for example, in an inert solvent such as benzene, methylene chloride, N, N-dimethylformamide or the like at 0 ° C. to reflux temperature, preferably 0 ° C. to room temperature, in the presence or absence of molecular sieve, pyridinium. Use chlorochromate or pyridinium dichromate,
Alternatively, dimethyl sulfoxide is used in the presence of oxalyl chloride and triethylamine in an inert solvent such as methylene chloride at −80 ° C. to room temperature, preferably −80 ° C. to 0 ° C., or in an inert solvent (for example, benzene). DCC is used in the presence of pyridine, trifluoroacetic acid, and dimethylsulfoxide in the presence or absence of coexistence or in the absence of coexistence, or at −10 ° C. in the presence or absence of an inert solvent (for example, benzene). To room temperature at room temperature in the presence of a tertiary amine such as triethylamine and dimethylsulfoxide, using a sulfur trioxide-pyridine complex.

Reaction (E) Of the compounds represented by the general formula [7], a compound in which X is a sulfur atom is oxidized by using a peracid such as m-chloroperbenzoic acid to obtain 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 ^{8 by} a method known in peptide chemistry.
Introduce the compound

[9] is obtained. Although various kinds of amino protecting groups R ⁸ can be considered, for example, R 8
^When a Boc group is used as ⁶ , the next reaction (

[9] →
It must be stable against acid treatment for de-Boc conversion in [10], and examples thereof include formyl group and trifluoroacetyl group which are acyl type protecting groups, and 9-fluorenyl which is urethane type protecting group. A methyloxycarbonyl group,
A methylsulfonylethyloxycarbonyl group and the like are preferable. 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 ⁸ 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 ⁸ from the compound represented by the general formula [10]. The method for removing the amino-protecting group R ⁸ varies depending on the kind of the protecting group, but a method known in peptide chemistry can be used. 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.

In the above description, each compound [5]
Or compound

An example of preparing compound [7] or compound [10] by subjecting [9] to condensation reaction with compound [6] has been shown, but this reaction may be carried out stepwise as follows. That is, as shown in the reaction (K), the general formula R ⁷ -B
A compound represented by —COOH (R ⁷ is a suitable amino-protecting group) is converted into a compound [5] in the same manner as in the above reaction (C).
A compound represented by the general formula [17] is obtained by performing a condensation reaction with Next, as shown in the reaction (L), the amino protecting group R ⁷ of the compound [17] is removed by a known method, and then the compound of the general formula W-U-A-H (A is -O-, -N) is used.
R ² - when) or the general formula W-U-A-COOH ( A is -CHR ¹ - it is possible to obtain a desired compound [7] by reaction with a compound represented by the case) of the.
A is -O in the former -, - NR ² - in the case of, for example, a W-U-A-H 1,4- dioxane, in a suitable solvent such as tetrahydrofuran, the presence of a tertiary amine such as triethylamine The reaction with phosgene, trichloromethyl chloroformate, carbonyldiimidazole and the like at -20 ° C to room temperature under the following conditions, and then the compound [1
This can be achieved by reacting with the deprotected form of [7]. When A is -NH-, W-U-N = C = O is added to a suitable solvent such as dichloromethane, chloroform, N, N-dimethylformamide in the presence of a tertiary amine such as triethylamine or It can also be achieved by reacting with a deprotected form of compound [17] in the absence of the compound at −20 ° C. to room temperature. 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 —CHR ¹ , the deprotected form of compound [17] can be converted into W—U—A—COOH or the corresponding acid chloride W—U by the method described in the above reaction (C). It is achieved by carrying out a condensation reaction with -A-COCl. Also with regard to the method for preparing the compound [10], the compound

From [9], in the same manner as in the above reaction (K), the general formula [1
The desired compound [10] can be prepared by carrying out the reaction (L) in the same manner as described above after the compound represented by 9] is prepared.

Further, the following general formula [1] (where X is-
SO-)

[Chemical 9] The optically active sulfinyl compound represented by can be produced according to the steps shown below.

[0019]

[Chemical 10] Each of the above reaction steps will be further described. Here, the symbols A, B, W, U and R are the same as above, and the reactions (C), (D), (K) and (L) are as described above.

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, te
It is carried out using rt-butyl hydroperoxide or cumene hydroperoxide.

Reaction (J) The optically active sulfoxide represented by the general formula [13] is treated with a base to give a corresponding carbanion, which is represented by the general formula [14] (R ⁹ represents a carboxyl protecting group). To an optically active sulfinyl compound represented by the general formula [1] or [16], respectively, by condensation with an aldehyde represented by the general formula [15] derived from the ester or compound [14] by a known method. I will get it. In this reaction, for example, optically active sulfoxide [13] is added to n-butyllithium at a reaction temperature of −78 ° C. to room temperature, preferably 0 ° C. or lower in an inert organic solvent such as THF or 1,4-dioxane. The corresponding carbanion is formed using a base such as lithium diisopropylamide and then the ester [14] or aldehyde [1]
5] is added and the condensation reaction is carried out at −78 ° C. to room temperature, preferably −78 ° C. to −20 ° C. Compounds used as synthetic raw material, N protected prolinal (2), HX-R [4], Q-COOH [6], ^{R 7} -
B-COOH, W-U-A-H, W-U-A-COO
H, CH ₃ —S—R [12] and ester of proline [18] are either available as known substances per se, or easily derived and synthesized from known precursors by known methods. can do. Further, when synthesizing the final target compound represented by the general formula [1], a protecting group is introduced at an appropriate stage as necessary, and an appropriate step (preferably the final step reaction (D) or reaction (J)) is performed. In the above), the protecting group may be removed.

Isolation and purification of the thus obtained compound represented by the general formula [1] from the reaction mixture are carried out by using any means commonly used in the field of synthetic organic chemistry. It can be isolated and purified by methods 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. To use the compound of the present invention as a medicine, usually,
It is administered systemically or locally, orally or parenterally. The dose varies depending on age, body weight, symptoms, therapeutic effect, administration method, etc., but is usually 1 mg to 10 per adult per dose.
It is orally administered once to several times daily in the range of 0 mg, or parenterally once to several times daily in the range of 0.2 mg to 20 mg per adult. 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 excipients, binders, lubricants, disintegrants, You may contain a solubilizing agent, a stabilizer, etc. The tablets or pills may optionally be coated with a film of gastric or enteric substance. Capsules include hard capsules and soft capsules. As a liquid composition for oral administration, a solution,
Emulsions, suspensions, syrups and elixirs are included. Such a liquid composition contains an inert diluent which is generally used, and additionally, an auxiliary agent such as a wetting agent and a suspending agent, a sweetening agent, a flavoring agent, an aromatic agent, a preservative. May be contained. 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 inert non-aqueous diluent and used, and if necessary, further preserved. Agents, wetting agents, emulsifiers,
It may contain an auxiliary agent such as a dispersant, a stabilizer and a solubilizing agent. 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. water,
Alternatively, it is used after adding a sterile injectable diluent.

[0023]

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- (phenoxyacetyl) -1- [N-
(Phenoxyacetyl) -L-prolyl] pyrrolidine (Compound 1) a) (2S) -1- (tert-butoxycarbonyl)
-2- (1,2-epoxyethyl) pyrrolidine sodium hydride (NaH content 60%, 1.55 g)
DMSO (20 ml) was added to and the mixture was stirred at 70 ° C. for 1 hour. The reaction solution was returned to room temperature, THF (20 ml) was added, the temperature was adjusted to -5 ° C, and a DMSO (30 ml) solution of trimethylsulfonium iodide (7.90 g) was added dropwise over 3 minutes. After stirring for 1 minute, tert-butoxycarbonyl-L-prolinal (5.13 g) in THF
The (15 ml) solution was added quickly and then stirred at room temperature for 1 hour. The reaction solution was poured into ice water (300 ml) and extracted with chloroform. The extract was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography (eluent: hexane-ethyl acetate) to obtain two diastereomers of the title compound, 3.12 g of low polar compound and 1.28 g of high polar compound (both epoxy compounds). It has not been decided about the three-dimensional part). In the following reaction, a low polar substance was used.

B) (2S) -1- (tert-butoxycarbonyl) -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (2S) -1- (tert-butoxycarbonyl) -2
-(1,2-epoxyethyl) pyrrolidine (1.50
g) in methanol (30 ml) in phenol (1.
50 g) and sodium methoxide (380 mg) were added, and the mixture was stirred and refluxed for 16 hours. The reaction mixture was concentrated, the residue was dissolved in ether, washed successively with 10% citric 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: hexane-ethyl acetate) to give the title compound (1.10 g).

C) (2S) -1- [N- (tert-butoxycarbonyl) -L-prolyl] -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (2S) -1- (tert-butoxycarbonyl) ) -2
-(1-Hydroxy-2-phenoxyethyl) pyrrolidine (1.93 g) was added to 4N hydrochloric acid-1,4-dioxane (3
1 ml) and stirred at room temperature for 0.5 hours. The residue obtained by concentrating and drying the reaction solution was dissolved in DMF (15 ml), and N- (tert-butoxycarbonyl) -L-proline (1.36 g) and N-methylmorpholine (0.69).
ml) and HOBt (1.02 g) were added. Reaction liquid
After cooling to 25 ° C., DCC (1.30 g) was added, and the reaction solution was stirred at −25 ° C. to 0 ° C. for 3 hours and then at room temperature for 16 hours. The precipitated dicyclohexylurea was filtered off, and the filtrate was concentrated. The residue was dissolved in ethyl acetate, washed successively 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 (2.10 g).

D) (2S) -2- (1-hydroxy-2)
-Phenoxyethyl) -1- [N- (phenoxyacetyl) -L-prolyl] pyrrolidine (2S) -1- [N- (tert-butoxycarbonyl) -L-prolyl] -2- (1-hydroxy-2-) Phenoxyethyl) pyrrolidine (996 mg) was dissolved in 4N hydrochloric acid-1,4-dioxane (12 ml) and 40
Stir for minutes. The reaction solution was concentrated and dried, the resulting residue was dissolved in methylene chloride (10 ml), triethylamine (0.68 ml) was added under ice cooling, and then phenoxyacetyl chloride (0.34 ml) was added dropwise. Under ice cooling 0.5
After stirring for 1.5 hours at room temperature and then for 1.5 hours, the reaction solution was poured into ice water and extracted with methylene chloride. The organic layer was washed successively 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 (1.05 g).

E) (2S) -2- (phenoxyacetyl) -1- [N- (phenoxyacetyl) -L-prolyl] pyrrolidine (2S) -2- (1-hydroxy-2-phenoxyethyl) -1- [N- (phenoxyacetyl) -L-prolyl] pyrrolidine (998 mg) was dissolved in DMSO (6 ml) and benzene (3 ml) to give pyridine (0.18 m).
1) and trifluoroacetic acid (0.09 ml) were added. After cooling the mixture with ice, DCC (941 mg) was added and the mixture was stirred at room temperature for 2.5 hours.
Stir for hours. The precipitated dicyclohexylurea was filtered off, and the filtrate was poured into ice-cooled 1N hydrochloric acid and extracted with ethyl acetate. The organic layer was washed successively with saturated aqueous sodium hydrogen carbonate and saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The residue was subjected to silica gel column chromatography (eluent: chloroform-
Purification with methanol) gave the title compound (759 mg).

Example 2 (2S) -2- (4-methoxyphenoxyacetyl)-
1- [N- (phenoxyacetyl) -L-prolyl] pyrrolidine (Compound 2) a) (2S) -1- (tert-butoxycarbonyl)
-2- [1-Hydroxy-2- (4-methoxyphenoxy) ethyl] pyrrolidine Prepared in Example 1 a) (2S) -1- (tert-
Butoxycarbonyl) -2- (1,2-epoxyethyl) pyrrolidine (11.70 g) in methanol (50 m
1) 4-methoxyphenol (13.60 g) and 1M sodium methoxide-methanol solution (54.9).
ml) was added and the mixture was stirred at 70 ° C. for 18 hours. The reaction solution was poured into saturated ammonium chloride water and extracted with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated. The residue was purified by silica gel column chromatography (eluent: hexane-ethyl acetate) to give the title compound (14.20 g).

B) (2S) -1- [N- (tert-butoxycarbonyl) -L-prolyl] -2- [1-hydroxy-2- (4-methoxyphenoxy) ethyl] pyrrolidine (2S) -1- (Tert-butoxycarbonyl) -2
-[1-hydroxy-2- (4-methoxyphenoxy)
Ethyl] pyrrolidine (1.57 g) was added to 4N hydrochloric acid-1,4
-Dissolved in dioxane (23 ml) and stirred at room temperature for 1 hour. The reaction solution was concentrated and dried, and the resulting residue was added to DMF (15
ml), N- (tert-butoxycarbonyl) -L-proline (1.00 g), N-methylmorpholine (0.51 ml), HOBt (0.75 g) and D.
Using CC (0.96 g), the condensation reaction was performed in the same manner as in the case of c) of Example 1 to give the title compound (1.41).
g) was obtained.

C) (2S) -2- [1-hydroxy-2
-(4-Methoxyphenoxy) ethyl] -1- [N-
(Phenoxyacetyl) -L-prolyl] pyrrolidine (2S) -1- [N- (tert-butoxycarbonyl) -L-prolyl] -2- [1-hydroxy-2-
(4-Methoxyphenoxy) ethyl] pyrrolidine (65
8 mg) was dissolved in 4N hydrochloric acid-1,4-dioxane (8 ml), and the mixture was stirred at room temperature for 0.5 hr. The residue obtained by concentrating and drying the reaction solution was dissolved in methylene chloride (10 ml), triethylamine (0.42 ml) was added under ice cooling, and then phenoxyacetyl chloride (0.21 ml).
Was dripped. After stirring under ice-cooling for 0.5 hours and then at room temperature for 2 hours, the reaction mixture was treated in the same manner as in Example 1, d) to obtain the title compound (531 mg).

D) (2S) -2- (4-methoxyphenoxyacetyl) -1- [N- (phenoxyacetyl)-
L-prolyl] pyrrolidine (2S) -2- [1-hydroxy-2- (4-methoxyphenoxy) ethyl] -1- [N- (phenoxyacetyl) -L-prolyl] pyrrolidine (478 mg) was DM
Dissolve in SO (3.5 ml) and benzene (1.5 ml)
And triethylamine (0.7 ml) were added. The mixture was ice-cooled, sulfur trioxide-pyridine complex (730 mg) was added little by little, and triethylamine (0.25 ml) was added to keep the reaction solution neutral. After stirring for 1 hour at 5 to 10 ° C, 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 (40
5 mg) was obtained.

Example 3 (2S) -2- (4-methoxyphenoxyacetyl)-
1- [N- (N-phenylglycyl) -L-prolyl]
Pyrrolidine (Compound 3) a) N- (tert-butoxycarbonyl) -N-phenylglycine N-phenylglycine (5.00 g) was dissolved in 1,4-dioxane-water (2: 1, 90 ml) and cooled under ice. 1N
Sodium hydroxide (33.1 ml) and di-tert-
Butyldicarbonate (7.95 g) was added, and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated, the residue was dissolved in 10% citric acid, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel column chromatography (eluent: chloroform-methanol-acetic acid) to give the title compound (8.30 g).

B) (2S) -1- {N- [N- (ter
t-butoxycarbonyl) -N-phenylglycyl]-
L-Prolyl} -2- [1-hydroxy-2- (4-methoxyphenoxy) ethyl] pyrrolidine (2S) -1- [N- (te
rt-Butoxycarbonyl) -L-prolyl] -2-
[1-Hydroxy-2- (4-methoxyphenoxy) ethyl] pyrrolidine (633 mg) was added to 4N hydrochloric acid-1,4-
It was dissolved in dioxane (6 ml) and stirred at room temperature for 0.5 hours. The reaction solution was concentrated and dried, and the residue and N- (te
rt-Butoxycarbonyl) -N-phenylglycine (340 mg) was dissolved in DMF (10 ml), N-methylmorpholine (0.15 ml) and HOBt (219).
mg) was added. The reaction solution was ice-cooled, water-soluble carbodiimide hydrochloride (263 mg) was added, and the mixture was stirred for 3 hours and then at room temperature overnight. The reaction solution was poured into ice water and extracted with ethyl acetate. The organic layer was washed successively with 10% citric acid and saturated aqueous sodium hydrogen carbonate,
The extract was dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel column chromatography (eluent: chloroform-methanol) to give the title compound (693 mg).
Got

C) (2S) -2- [1-hydroxy-2
-(4-Methoxyphenoxy) ethyl] -1- [N-
(N-Phenylglycyl) -L-prolyl] pyrrolidine (2S) -1- {N- [N- (tert-butoxycarbonyl) -N-phenylglycyl] -L-prolyl}-
2- [1-hydroxy-2- (4-methoxyphenoxy) ethyl] pyrrolidine (656 mg) was added to 4N hydrochloric acid-
It was dissolved in 1,4-dioxane (5 ml) and stirred at room temperature for 0.5 hours. The residue obtained by concentrating the reaction solution was dissolved in saturated aqueous sodium hydrogen carbonate and extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate and concentrated to give the title compound (518 mg).

D) (2S) -2- (4-methoxyphenoxyacetyl) -1- [N- (N-phenylglycyl)
-L-prolyl] pyrrolidine (2S) -2- [1-hydroxy-2- (4-methoxyphenoxy) ethyl] -1- [N- (N-phenylglycyl) -L-prolyl] pyrrolidine (488 mg) D
MSO (3 ml), benzene (1.5 ml), triethylamine (0.73 ml + 0.15 ml) and sulfur trioxide-pyridine complex (778 mg) were reacted and treated in the same manner as in the case of Example 2 d). A compound (288 mg) was obtained.

Example 4 (2S) -2- (4-methoxyphenoxyacetyl)-
1- {N-[(phenylthio) acetyl] -L-prolyl} pyrrolidine (Compound 4) a) (2S) -2- [1-hydroxy-2- (4-methoxyphenoxy) ethyl] -1- {N- [(Phenylthio) acetyl] -L-prolyl} pyrrolidine (2S) -1- [N- (te
rt-Butoxycarbonyl) -L-prolyl] -2-
[1-Hydroxy-2- (4-methoxyphenoxy) ethyl] pyrrolidine (622 mg) was added to 4N hydrochloric acid-1,4-.
It was dissolved in dioxane (6 ml) and stirred at room temperature for 0.5 hours. The reaction solution was concentrated and dried, and the resulting residue was dissolved in methylene chloride (15 ml) and DMF (5 ml) and cooled under ice-cooling to N.
-Methylmorpholine (0.14 ml), (phenylthio) acetic acid (210 mg), HOBt (190 mg) and water-soluble carbodiimide hydrochloride (242 mg) were added, and the mixture was added to 0.2.
Stir for 5 hours. Then, after stirring at room temperature overnight, the reaction mixture was treated in the same manner as in Example 3, b) to give the title compound (6
64 mg) was obtained.

B) (2S) -2- (4-methoxyphenoxyacetyl) -1- {N-[(phenylthio) acetyl] -L-prolyl} pyrrolidine (2S) -2- [1-hydroxy-2- ( 4-Methoxyphenoxy) ethyl] -1- {N-[(phenylthio)
Acetyl] -L-prolyl} pyrrolidine (620 mg)
Was treated with DMSO (10 ml), benzene (5 ml), triethylamine (0.90 ml) and sulfur trioxide-pyridine complex (960 mg) in the same manner as in the case of d) of Example 2 to treat the title compound ( 510m
g) was obtained.

Example 5 (2S) -1- {N-[(1-adamantyl) methoxycarbonyl] -L-prolyl} -2- (4-methoxyphenoxyacetyl) pyrrolidine (Compound 5) a) N-[( 1-adamantyl) methoxycarbonyl]
-L-proline benzyl ester trichloromethyl chloroformate (0.36 ml)
In THF solution (40 ml) under ice-cooling, 1-adamantanemethanol (1.00 g) and triethylamine (0.
A THF solution (20 ml) containing 84 ml) was added dropwise.
After stirring at room temperature for 1.5 hours, a solution of L-proline benzyl ester (1.45 g) in methylene chloride (15 m
1) was added, and then triethylamine (1.7 ml) was added dropwise. After stirring at room temperature for 20 minutes, the reaction solution was poured into water and extracted with ethyl acetate. The organic layer was washed successively with 1N hydrochloric acid and saturated aqueous sodium hydrogen carbonate, dried over anhydrous magnesium silicate and concentrated.
The residue was subjected to silica gel column chromatography (eluent:
Purification by hexane-ethyl acetate) gave the title compound (2.10 g).

B) N-[(1-adamantyl) methoxycarbonyl] -L-proline N-[(1-adamantyl) methoxycarbonyl] -L
-Proline benzyl ester (1.98 g) in methanol (60 ml)
10% Pd-C (0.20 g) was added to and the mixture was stirred at room temperature under a hydrogen atmosphere for 1.5 hours. After removing the catalyst by filtration, the filtrate was concentrated to give the title compound (1.53 g).

C) (2S) -1- {N-[(1-adamantyl) methoxycarbonyl] -L-prolyl} -2-
[1-Hydroxy-2- (4-methoxyphenoxy) ethyl] pyrrolidine Prepared in Example 2 a) (2S) -1- (tert-
Butoxycarbonyl) -2- [1-hydroxy-2-
(4-Methoxyphenoxy) ethyl] pyrrolidine (1.
40 g) to 4N hydrochloric acid-1,4-dioxane (40 ml)
And was stirred at room temperature for 0.5 hour, and then the reaction solution was concentrated to dryness. The residue is treated with N-in the same manner as in Example 3, b).
The title compound (1.56 g) was obtained by subjecting it to a dehydration condensation reaction with [(1-adamantyl) methoxycarbonyl] -L-proline (1.16 g).

D) (2S) -1- {N-[(1-adamantyl) methoxycarbonyl] -L-prolyl} -2-
(4-Methoxyphenoxyacetyl) pyrrolidine (2S) -1- {N-[(1-adamantyl) methoxycarbonyl] -L-prolyl} -2- [1-hydroxy-2- (4-methoxyphenoxy) ethyl] pyrrolidine (1.40 g) was oxidized with a sulfur trioxide-pyridine complex in the same manner as in Example 2 d) to obtain the title compound (0.98 g).

Example 6 (2S) -1- [N- (benzylaminocarbonyl)-
L-thioprolyl] -2- (phenoxyacetyl) pyrrolidine (Compound 6) a) (2S) -1- [N- (tert-butoxycarbonyl) -L-thioprolyl] -2- (1-hydroxy-
2-Phenoxyethyl) pyrrolidine (2S) -1- (tert-) prepared in Example 1 b).
Butoxycarbonyl) -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (3.00 g) was dissolved in 4N hydrochloric acid-1,4-dioxane (30 ml), stirred at room temperature for 0.5 hours, and then the reaction solution was concentrated. Dried up. The residue was treated with N- (tert-butoxycarbonyl) -L-thioproline (2.50 g) as in Example 3b).
The title compound (2.49 g) was obtained by dehydration condensation with.

B) (2S) -1- [N- (benzylaminocarbonyl) -L-thioprolyl] -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (2S) -1- [N- (tert- Butoxycarbonyl) -L-thioprolyl] -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (2.47 g) was dissolved in 4N hydrochloric acid-1,4-dioxane (15 ml) and stirred at room temperature for 0.5 hours. rear,
The reaction solution was concentrated to dryness. Methylene chloride solution of the residue (50
ml) under ice-cooling, triethylamine (0.90 ml)
And benzyl isocyanate (0.80 ml) were added dropwise, and the mixture was stirred for 1.5 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 (1.95 g).

C) (2S) -1- [N- (benzylaminocarbonyl) -L-thioprolyl] -2- (phenoxyacetyl) pyrrolidine (2S) -1- [N- (benzylaminocarbonyl)-
L-thioprolyl] -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (1.80 g) was prepared according to Example 2
It was oxidized in the same manner as in the case of d) above using a sulfur trioxide-pyridine complex to obtain the title compound (1.40 g).

Example 7 (2S) -1- {N- [N-benzyl-N- (ethoxycarbonylmethyl) aminocarbonyl] -L-prolyl} -2- (phenoxyacetyl) pyrrolidine (Compound 7) a) ( 2S) -1- {N- [N-benzyl-N- (ethoxycarbonylmethyl) aminocarbonyl] -L-prolyl} -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine In the case of Example 5 a) Similarly to, trichloromethyl chloroformate (0.37 ml), N-benzylglycine ethyl ester (1.2 ml) and Example 1.
(2S) -1- [N- (tert-butoxycarbonyl) -L-prolyl] -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine prepared in step c) above was treated with hydrochloric acid to obtain (2S). The title compound (2.12 g) was obtained using 2- (1-hydroxy-2-phenoxyethyl) -1- (L-prolyl) pyrrolidine hydrochloride (1.75 g).

B) (2S) -1- {N- [N-benzyl-N- (ethoxycarbonylmethyl) aminocarbonyl] -L-prolyl} -2- (phenoxyacetyl) pyrrolidine (2S) -1- {N -[N-benzyl-N-
(Ethoxycarbonylmethyl) aminocarbonyl] -L
-Prolyl} -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine (2.00 g), Example 2d).
In the same manner as in (2), the title compound (1.25 g) was obtained by oxidation with a sulfur trioxide-pyridine complex.

Example 8 (2S) -1- [N-[[(S) -α- (ethoxycarbonyl) benzylamino] carbonyl] -L-prolyl] -2- (phenoxyacetyle) pyrrolidine (Compound 8) a) (2S) -1- [N-[[(S) -α- (ethoxycarbonyl) benzylamino] carbonyl] -L-prolyl] -2- (1-hydroxy-2-phenoxyethyl) pyrrolidine Example 5 In the same manner as in the case a), trichloromethyl chloroformate (0.38 ml), L-phenylglycine ethyl ester hydrochloride (1.37 g) and (2S) -2- (1-hydroxy-2-phenoxyethyl). ) -1- (L-Prolyl) pyrrolidine hydrochloride (1.8
0 g) to give the title compound (1.65 g).

B) (2S) -1- [N-[[(S) -α
-(Ethoxycarbonyl) benzylamino] carbonyl] -L-prolyl] -2- (phenoxyacetyl) pyrrolidine (2S) -1- [N-[[(S) -α- (ethoxycarbonyl) benzylamino] carbonyl]- L-prolyl] -2- (1-hydroxy-2-phenoxyethyl)
Pyrrolidine (1.63 g) was oxidized with a sulfur trioxide-pyridine complex in the same manner as in the case of d) of Example 2 to obtain the title compound (0.95 g).

Example 9 (2S) -1- [N- (benzylaminocarbonyl)-
L-prolyl] -2-[(2-pyridyloxy) acetyl] pyrrolidine (Compound 9) a) (2S) -1- (tert-butoxycarbonyl)
-2- [1-hydroxy-2- (2-pyridyloxy)
Ethyl] pyrrolidine In the same manner as in Example 2 a), (2S) -1-
A crude product of the title compound from (tert-butoxycarbonyl) -2- (1,2-epoxyethyl) pyrrolidine (1.52 g) and 2-pyridinol (1.35 g) (1.
60 g) was obtained.

B) (2S) -1- [N- (tert-butoxycarbonyl) -L-prolyl] -2- [1-hydroxy-2- (2-pyridyloxy)
Ethyl] pyrrolidine (2S) -1- (tert-butoxycarbonyl) -2
-[1-Hydroxy-2- (2-pyridyloxy) ethyl] pyrrolidine (1.54 g) was dissolved in 4N hydrochloric acid-1,4-dioxane (40 ml), and the mixture was stirred at room temperature for 0.5 hr. The residue obtained by concentrating and drying the reaction solution is subjected to a condensation reaction with N- (tert-butoxycarbonyl) -L-proline (1.10 g) in the same manner as in b) of Example 3. Gave the title compound (0.83 g).

C) of (2S) -1- [N- (benzylaminocarbonyl) -L-prolyl] -2- [1-hydroxy-2- (2-pyridyloxy) ethyl] pyrrolidine of Example 6, b). Similarly to the case, (2S) -1-
The title compound (670 mg) was obtained from [N- (tert-butoxycarbonyl) -L-prolyl] -2- [1-hydroxy-2- (2-pyridyloxy) ethyl] pyrrolidine (830 mg).

D) (2S) -1- [N- (benzylaminocarbonyl) -L-prolyl] -2-[(2-pyridyloxy) acetyl] pyrrolidine (2S) -1- [N- (benzylaminocarbonyl) ) −
L-prolyl] -2- [1-hydroxy-2- (2-pyridyloxy) ethyl] pyrrolidine (600 mg),
Oxidation with sulfur trioxide-pyridine complex in the same manner as in Example 2 d) gave the title compound (560 mg).
Got

Example 10 (2S) -2- (phenoxyacetyl) -1- {N-
[3- (1-Piperidinyl) propionyl] -L-prolyl} pyrrolidine (Compound 10) a) (2S) -2- (1-hydroxy-2-phenoxyethyl) -1- {N- [3- (1- Piperidinyl) propionyl] -L-prolyl} pyrrolidine (2S) -1- [N- (te
rt-Butoxycarbonyl) -L-prolyl] -2-
(1-Hydroxy-2-phenoxyethyl) pyrrolidine (1.46 g) was added to 4N hydrochloric acid-1,4-dioxane (10
ml) and left for 15 minutes at room temperature. The residue obtained by concentrating and drying the reaction solution was treated with 1-piperidinepropionic acid (0.63 g) in the same manner as in Example 3, b).
By subjecting it to a condensation reaction with the title compound (1.40
g) was obtained.

B) (2S) -2- (phenoxyacetyl) -1- {N- [3- (1-piperidinyl) propionyl] -L-prolyl} pyrrolidine (2S) -2- (1-hydroxy-2-) Phenoxyethyl) -1- {N- [3- (1-piperidinyl) propionyl] -L-prolyl} pyrrolidine (1.40 g)
Oxidation with sulfur trioxide-pyridine complex in the same manner as in Example 2 d) gave the title compound (0.30 g).
Got

Example 11 (2S) -2- (phenoxyacetyl) -1- {N-
[3-phenyl-2 (S)-(1-piperidinyl) bropionyl] -L-prolyl} pyrrolidine (Compound 11) a) (2S) -3-phenyl-2- (1-piperidinyl) propionic acid tert-butyl ester Potassium carbonate (5.38 g) and 1,5-dibromopentane (2.98 g) were added to a DMF solution (50 ml) of L-phenylalanine tert-butyl ester hydrochloride (3.05 g), and the mixture was added at 80 ° C. for 2 hours and then at room temperature. In 2
Stir for 2 hours. Dilute the reaction solution with diethyl ether,
The extract was washed successively with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (eluent: methylene chloride-methanol) to give the title compound (1.24 g).

B) (2S) -3-phenyl-2- (1-
Piperidinyl) propionic acid hydrochloride (2S) -3-phenyl-2- (1-piperidinyl) propionic acid tert-butyl ester (800 m
g) was dissolved in trifluoroacetic acid (5 ml) and stirred at room temperature for 14
Stir for hours. After concentrating the reaction solution, the residue was treated with 4N hydrochloric acid-1,
It was dissolved in 4-dioxane (3 ml) and concentrated. Diethyl ether was added to the residue, and the precipitated crystals were collected by filtration to give the title compound (524 mg).

C) (2S) -2- (1-hydroxy-2)
-Phenoxyethyl) -1- {N- [3-phenyl-2
(S)-(1-Piperidinyl) propionyl] -L-prolyl} pyrrolidine Prepared in (2S) -1- [N- (tert) prepared in c) of Example 1 in the same manner as in b) of Example 3. -Butoxycarbonyl) -L-prolyl] -2- (1-hydroxy-
2-phenoxyethyl) pyrrolidine (751 mg) was added to 4
The residue obtained by treatment with N hydrochloric acid was (2S) -3-phenyl-2.
-(1-Piperidinyl) propionohydrochloride (500 m
g) by subjecting it to a condensation reaction with the title compound (818
mg) was obtained.

D) (2S) -2- (phenoxyacetyl) -1- {N- [3-phenyl-2 (S)-(1-piperidinyl) propionyl] -L-prolyl} pyrrolidine (2S) -2- (1-Hydroxy-2-phenoxyethyl) -1- {N- [3-phenyl-2 (S)-(1-piperidinyl) propionyl] -L-prolyl} pyrrolidine (818 mg) was added to Example 2d). Oxidation using a sulfur trioxide-pyridine complex was carried out in the same manner as in the above to give the title compound (582 mg). These compounds 1 to 11
The physicochemical properties of are shown in Tables 1 to 3.

[0060]

[Table 1]

[0061]

[Table 2]

[0062]

[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.

[0063]

[Table 4]

Then, the proline derivative of the above-mentioned general formula [1] according to the present invention was tested for its prolyl endopeptidase inhibitory activity in vitro and its inhibitory activity against various proteolytic enzymes. 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 / 1, pH 6.8, 1 mM dithiothreitol and 0.
Contains 5 mM EDTA. J. Neurohem. ，
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 Co., Ltd.). Was added and 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 An experiment using 0.1 M potassium-sodium phosphate buffer (pH 7.0) was performed, and
The fluorescence intensities a ₂ and a ₃ were measured (protein nucleic acid enzyme,
29 , 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 the table.

[0065]

[Equation 1]

[0066]

[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 inhibitory activity against various proteolytic enzymes at a predetermined concentration. It was found to specifically inhibit peptidases.

[0068]

[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), 0.2 μM of chymotrypsin (derived from bovine pancreas, manufactured by Sigma) as an enzyme solution, and 7- (N-succinyl-leucyl-leucyl-valyl-tyrosyl) -4- as a substrate solution. Using a 200 μM buffer solution of methylcoumarinamide (manufactured by Peptide Laboratories, Inc.) 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
A solution of elastase (derived from porcine pancreas, manufactured by Sigma) at 0.2 μM in the same solution as an enzyme solution was used as a substrate solution, and 7- (N-succinyl-alanyl-prolyl-) was used as a substrate solution.
Using a 200 μM buffer solution of alanyl) -4-methylcoumarinamide (manufactured by Peptide Research Co., Ltd.), 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 (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

[0074]

[Equation 2]

[0075]

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 (72) Inventor Katsuo Toide 23 Furuki, Hadano City, Kanagawa Prefecture Japan Tobacco Inc. Safety Research Institute (72) Inventor Itaro Uchida 6-2 Umegaoka, Midori-ku, Yokohama, Japan Tobacco Inc. Pharmaceutical Research Institute

Claims (1)

[Claims] 1. A general formula [1]: [In the formula, A is —O—, —CHR ¹ — or —NR ² — (wherein R ¹ is a hydrogen atom or a heterocycle, and R ² is a lower alkoxycarbonyl lower alkyl group); B
Is Or —NR ³ —CHR ⁴ — (wherein R ³ and R ^{4 are}
Are the same or different and are a hydrogen atom or a lower alkyl group); W is a phenyl group which may be substituted with a halogen atom, a lower alkyl group or a lower alkoxy group, an adamantyl group, a lower alkyl group or a heterocycle; U
Is —O—, —S—, —NH— or —CHR ⁵ — (wherein R ⁵ is a hydrogen atom or a lower alkoxycarbonyl group); X is —S—, —SO—, —SO ₂ —. , -O
-Or -NH-; R is (Where D is a benzene ring or a heterocycle, and Y and Z
Are the same or different and are a lower alkyl group, an amino group, a nitro group, a hydroxyl group or a lower alkoxy group which may be substituted with a hydrogen atom, a halogen atom or a fluorine atom). However, when U is —CH ₂ —, at least W is an adamantyl group or a heterocycle, or A is —CHR ¹ — or —NR ² — (wherein R ¹ is a heterocycle, R is ² is a lower alkoxycarbonyl lower alkyl group), B is thiazolidine, or R means either a heterocycle].