JPH0776211B2 - Pyroglutamic acid derivative - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a pyroglutamic acid derivative useful as a drug.

More specifically, the present invention has a prolyl endopeptidase (hereinafter referred to as PEP) inhibitory activity, and has a general formula useful as a therapeutic agent for amnesia. (In the formula, R is a 5-membered nitrogen-containing saturated heterocyclic group which may contain a sulfur atom, X is a single bond or -CO-, and Y is -O- or -NH-. , N is an integer of 1 to 2) and a pyroglutamic acid derivative and a pharmaceutically acceptable salt thereof.

[Conventional technology]

With the aging of the population, the problem of geriatric medicine is being emphasized. Among them, senile dementia is a serious social problem, and therefore, the immediate development of an effective therapeutic agent is desired.

Hitherto, as therapeutic agents for amnesia and dementia, there have been used cerebral circulation improving agents due to cerebral vasodilatory action, cerebral metabolism activating agents due to cerebral cell oxygen consumption enhancing action and the like. However, although these agents are effective for dementia due to cerebrovascular disorder, it has been considered difficult to be surely effective for dementia due to other causes.

PEP is known as an enzyme that acts on a physiologically active peptide containing proline and a synthetic substrate to specifically cleave the carboxyl side of proline. This enzyme is associated with memory vasopressin (Vasopressin) and thyrotropin releasing hormone (Thyrotropin Releasing Hormone, TR
H) and the like are decomposed, various studies have been conducted on the relationship between the inhibitory activity of this enzyme and the anti-amnestic effect, and as a result,
It has been suggested that a PEP inhibitor may be a therapeutic agent for dementia and amnesia (Biochemistry, 55, No. 8, page 831, 1983.
Year).

[Problems to be Solved by the Invention]

Since the cerebral circulation improving agent and the cerebral metabolism activating agent which have been conventionally used as a therapeutic agent for amnesia and dementia are not so effective, development of a therapeutic agent for amnesia with a new action has been desired.

As a result of investigations to find out a therapeutic agent for amnesia having an action different from that of conventional therapeutic agents, the present inventors have found that a certain type of pyroglutamic acid derivative exhibits a strong PEP inhibitory activity and can achieve the object.

The present invention is based on these findings.

[Means for Solving the Problems]

The pyroglutamic acid derivative represented by the above general formula (I) of the present invention has a strong PEP inhibitory activity and low toxicity, and is useful as a therapeutic agent for amnesia.

In the above formula (I) of the present invention, R is a 5-membered nitrogen-containing saturated heterocyclic group which may contain sulfur, such as 4-thiazolidinyl, 2-pyrrolidinyl and 5-oxo-2-pyrrolidinyl group. Can be given.

The compound of the general formula (I) of the present invention is a novel compound and can be produced as follows. For example,
General formula (Wherein R has the same meaning as described above), a carboxylic acid represented by the general formula (A in the formula is a hydroxyl group or an amino group, and X and n
Has the same meaning as described above).

General formula (Wherein X, R and n have the same meanings as described above), the compound represented by the general formula (II) (Wherein B is a halogen atom, X and n have the same meanings as described above), and a compound such as
Reacting in the presence of sodium hydrogen carbonate, sodium hydroxide, or alternatively the general formula A compound represented by the formula (wherein X and n have the same meanings as described above) and a carboxylic acid represented by the general formula R-COOH (VI) (wherein R has the same meanings as described above) It can be produced by reacting a reactive functional derivative in the presence of a base such as sodium hydrogen carbonate or sodium hydroxide.

In producing the compound of the general formula (I) of the present invention, when the compound represented by the general formula (II) is reacted with the compound represented by the general formula (III), the reaction is carried out in the presence of a condensing agent. As such a condensing agent, a condensing agent generally used in peptide synthesis, such as N, N'-dicyclohexylcarbodiimide, is used.

When the compound represented by the general formula (II) or (VI) used in the method for producing the compound of the general formula (I) or (Ia) of the present invention is required to protect the amino group, The desired compound (I) is prepared by protecting the amino group with a suitable amino-protecting group, for example, t-butoxycarbonyl group, followed by reaction, and then removing the amino-protecting group.
Or get (Ia).

Examples of the reactive functional derivative of the compound of the general formula (VI) used in the method for producing the compound of the general formula (Ia) of the present invention include active ester, acid anhydride, mixed acid anhydride and the like.

The compound represented by the general formula (I) of the present invention can be converted into a pharmaceutically acceptable acid addition salt according to a conventional method,
Examples of these salts include hydrochloride, sulfonate, p-toluenesulfonate, tartrate, and fumarate.

The compound of the general formula (I) of the present invention has 1 to 2 asymmetric carbons including a pyroglutamic acid moiety, but in the present invention, the configuration of the substituent on each asymmetric carbon is either R or S. However, it may be a mixture thereof. Each optically active compound can be obtained by using an optically active compound as a starting material and condensing sterically.

The compound of general formula (I) of the present invention can be made into various pharmaceutical preparations according to a conventional method. That is, if necessary, pharmaceutical additives such as an excipient, a disintegrant, a condensing agent, a lubricant, etc. are added, and various preparations are prepared according to a conventional method, for example, tablets, powders, granules, capsules, etc. Can be

When the compound of the general formula (I) of the present invention is used as a therapeutic agent for amnesia, its dose is appropriately determined according to the age, weight, sex, degree of symptoms of the patient, etc. 50 to 1000 mg for parenteral, 1 to 50 for parenteral administration
Used in the 0 mg range.

〔The invention's effect〕

The compound of the general formula (I) of the present invention is a bovine brain using N-carbobenzooxy-L-glycyl-L-prolyl-β-naphthylamide (hereinafter referred to as Z-Gly-Pro-β-NA) as a substrate. in inhibitory activity measuring test on derived prolyl endopeptidase, generally at 6 × 10 ^-4 ~8 × 10 ^-4 molar concentration 50
Shows% inhibitory activity.

Preferably, N-{(R)-(-)-thiazolidine-4
-Carbonyl} -phenacyl pyroglutamate-hydrochloride having an IC ₅₀ value of 6.5 x 10 ^-4 mol. As described above, the compound of the general formula (I) of the present invention has a strong PEP inhibitory activity and low toxicity, and is therefore a safe and excellent compound useful as a therapeutic agent for amnesia.

〔Example〕

The following reference examples and examples are given to describe the present invention in more detail. The melting points of the compounds in Reference Examples and Examples are all uncorrected.

Reference Example 1 (R)-(−)-Nt-butoxycarbonyl-thiazolidine-4-carboxylic acid (R)-(−)-thiazolidine-4-carboxylic acid 13.3 g
And triethylamine 14 ml with dioxane 50 ml and water
It was dissolved in 50 ml of a mixed solvent, 24 g of di-t-butyl-dicarbonate was added under ice cooling, and the mixture was stirred at room temperature for 20 hours. 100 ml of water was added to the reaction solution, which was washed with ethyl acetate, and 10% aqueous citric acid solution was added under ice-cooling until the aqueous layer reached pH 2. The mixture was extracted with ethyl acetate, the ethyl acetate layer was washed with saturated brine and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain 21.6 g (93%) of the desired product.

IR (KBr): ν _co 1745,1630cm ^-1 NMR (CDCl ₃ ) δ: 1.48 (s, 9H), 3.30 (s, 2H), 4.35 ~ 4.95 (m, 3H), 10.10 (br-s, 1H) Reference Example 2 (S) -N-benzyloxycarbonyl-pyroglutamic acid N-hydroxysuccinimide ester Carbobenzoxy-L-pyroglutamic acid (8.2 g) was dried with N,
It was dissolved in 60 ml of N-dimethylformamide, 3.6 g of N-hydroxysuccinimide and 6.5 g of N, N'-dicyclohexylcarbodiimide were added, and the mixture was stirred at room temperature overnight.
After completion of the reaction, the solvent was distilled off under reduced pressure, ethyl acetate was added, and the insoluble material was filtered off. The filtrate was evaporated under reduced pressure and washed with ether to obtain 10.4 g of the desired product.

Melting point: 131-134 ° C IR (KBr): ν _co 1820,1785,1735cm ^-1 NMR (CDCl ₃ ) δ: 2.30-2.85 (br-m, 4H), 2.87 (s, 4H), 4.98 (dd, 1H) ), 5.32 (dd, 2H), 7.30 to 7.50 (m, 5H) Elemental analysis value: (as C ₁₇ H ₁₆ N ₂ O ₇ ) C% H% N% Calculated value 56.67 4.48 7.77 Measured value 56.83 4.61 7.84 Reference example 3 Using (R)-(−)-Nt-butoxycarbonyl-thiazolidine-4-carboxylic acid or (S) -Nt-butoxycarbonyl-proline in place of carbobenzoxy-L-pyroglutamic acid, The following compounds were produced by the same method as in Reference Example 2.

(R)-(−)-Nt-butoxycarbonyl-thiazolidine-4-carboxylic acid N-hydroxysuccinimide ester IR (KBr): ν _co 1820, 1785, 1745, 1695 cm ⁻¹ NMR (CDCl ₃ ). δ: 1.49 (s, 9H), 2.84 (s, 4H), 3.30 to 3.60 (m, 2H), 4.40 to 4.70 (m, 2H), 4.80 to 5.00 (m, 1H) (S) -Nt- Butoxycarbonyl-proline N-hydroxysuccinimide ester IR (KBr): ν _co 1820,1790,1750, 1700 cm ^-1 NMR (CDCl ₃ ) δ: 1.48 (s, 9H), 1.85 to 2.15 (m, 2H) , 2.25 ~ 2.50 (m, 2H), 2.83 (s, 4H), 3.35 ~ 3.65 (m, 2H), 4.55 (dd, 1H) Reference Example 4 Benzyl pyroglutamate Pyroglutamate 2.0 g dried N, N-dimethylformamide Dissolve in 10 ml, add 1.3 g of sodium hydrogen carbonate,
The mixture was stirred at 50 ° C for 1 hour. To the reaction solution was added benzyl chloride (1.78 ml) at room temperature, and the mixture was stirred at 60 ° C overnight. After completion of the reaction, the solvent was distilled off under reduced pressure, water was added to the residue, the mixture was extracted with ethyl acetate, washed with water and saturated saline, and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain an oily substance, ether was added, and the precipitated crystals were collected by filtration. Further, it was recrystallized from ethanol-hexane to obtain 2.35 g of the desired product.

Melting point: 58 to 63 ° C IR (KBr): ν _co 1735,1695 cm ^-1 NMR (CDCl ₃ ) δ: 2.20 to 2.60 (m, 4H), 4.25 to 4.35 (m, 1H), 5.20 (s, 2H) , 5.80 to 5.95 (br-s, 1H), 7.30 to 7.45 (m, 5H) Elemental analysis value: (as C ₁₂ H ₁₃ O ₃ N) C% H% N% Calculated value 65.75 5.94 6.39 Measured value 65.77 6.02 6.52 Reference Example 5 The following compound was produced in the same manner as in Reference Example 4 using phenethyl bromide instead of benzyl chloride.

Phenethyl Pyroglutamate Melting point: 75-76 ° C IR (KBr): ν _co 1735,1680 cm ^-1 NMR (CDCl ₃ ) δ: 2.05-2.50 (m, 4H), 3.00 (t, 2H), 4.15-4.25 (m , 1H), 4.39 (dt, 2H), 6.18 (s, 1H), 7.10 to 7.40 (m, 5H) Example 1 benzyl N-{(R)-(-)-thiazolidine-4-carbonyl} -pyroglutamate Hydrochloride (Compound A) Benzyl pyroglutamate (1.5 g) was dissolved in dry benzene (45 ml), and sodium hydride (oil-based) (300 mg) was added under ice-cooling stirring, and the stirring was continued for a while. (R)-in the above suspension
(-)-N-t-butoxycarbonyl-thiazolidine-
4-carboxylic acid N-hydroxysuccinimide ester
2.26 g was added and the mixture was stirred at room temperature for 14 hours. The reaction solution was washed with water and dried over anhydrous magnesium sulfate to give 3.31 as an oil.
got g. The oily matter was purified by silica gel column chromatography (eluting solvent: chloroform), and N-
{(R)-(-)-Nt-Butosicarbonyl-thiazolidine-4-carbonyl} -benzyl pyroglutamate
1.94 g was obtained.

Rf value: 0.36 (developing solvent: chloroform / ethanol = 50/1) IR (neat): ν _co 1750,1700 cm ^-1 MS: MH ⁺ , 435 NMR (CDCl ₃ ) δ: 1.41,1.46 (s, s , 9H isomer), 2.00 ~ 2.90 (m, 4H), 2.95 ~ 3.10 (m, 1H), 3.45 ~ 3.65 (m, 1H), 4.40 ~ 4.75 (m, 2H), 4.77, 4.89 (dd, d, 1H), 5.05 to 5.30 (m, 2H), 5.65 to 5.90 (m, 1H), 7.30 to 7.45 (m, 5H) Dissolve 160 mg of the above benzyl ester compound in dry ethyl acetate, and dry under ice cooling to dry hydrogen chloride. Gas was introduced and stirred for 75 minutes. The reaction solution was distilled off under reduced pressure to obtain 162 mg of the desired product.
Got

IR (KBr): ν _co 1750,1700 cm ^-1 MS: MH ⁺ -HCl, 335 NMR (CDCl ₃ ) δ: 2.00 ~ 2.80 (m, 4H), 3.10 ~ 3.35 (m, 1H), 3.60 ~ 3.75 ( m, 1H), 4.40 to 4.50 (m, 1H), 4.60 to 4.70 (m, 1H), 4.80 to 5.10 (m, 1H), 5.10 to 5.30 (m, 2H), 5.50 to 5.65 (m, 1H), 7.30 to 7.45 (m, 5H) Example 2 The following compounds were produced in the same manner as in Example 1 using phenethyl pyroglutamate instead of benzyl pyroglutamate.

N-{(R)-(−)-thiazolidine-4-carbonyl} -phenethyl pyroglutamate hydrochloride (Compound B) IR (KBr): ν _co 1745,1700 cm ⁻¹ MS: MH ⁺ —HCl, 349 NMR (CDCl ₃ ) δ: 1.85 to 2.00 (m, 1H), 2.10 to 2.70 (m, 3H), 2.80 to 3.30 (m, 3H), 3.55 to 3.75 (m, 1H), 4.30 to 4.95 (m, 5H) , 5.40 to 5.65 (m, 1H), 7.15 to 7.40 (m, 5H) Example 3 (R)-(-)-Nt-butoxycarbonyl-thiazolidine-4-carboxylic acid N-hydroxysuccinimide ester Instead of (S) -Nt-butoxycarbonyl-proline N-hydroxysuccinimide ester, the following compound was produced in the same manner as in Example 1.

N-{(S) -Prolyl} -benzyl pyroglutamate hydrochloride (Compound C) IR (KBr): ν _co 1750,1700 cm ^-1 MS: MH ⁺ -HCl, 317 NMR (CDCl ₃ ) δ: 1.70 ~ 2.80 (m, 8H), 3.48 (br-s, 2H), 4.70 to 5.40 (m, 2H), 5.16 (q, 2H), 7.30 to 7.45 (m, 5H) Example 4 N-{(R)- (-)-Thiazolidine-4-carbonyl} -pyroglutamic acid benzylamide hydrochloride (Compound D) N-{(R) -Nt-butoxycarbonyl-thiazolidine-4-carbonyl} -benzyl pyroglutamate 72
Dissolve 0 mg in 60 ml ethanol and use 10% palladium-carbon.
1.46 g was added, and the mixture was stirred at room temperature for 24 hours. After the reaction,
The reaction solution was filtered, the filtrate was evaporated under reduced pressure, and N-{(R)
470 mg of -Nt-butoxycarbonyl-thiazolidine-4-carbonyl} -pyroglutamic acid was obtained.

IR (neat): ν _co 1740,1700 cm ^-1 NMR (DMSO) δ: 1.32,1.41 (s, s, 9H isomer), 1.90 to 2.10 (m, 1H), 2.30 to 3.20 (m, 5H), 4.30 to 4.70 (m, 3H), 5.55 to 5.75 (m, 1H) The above-mentioned pyroglutamic acid compound (470 mg) was dissolved in dry acetonitrile (20 ml), and benzylamine (160 ml), 1-hydroxybenzotriazole monohydrate (210 mg) and N, N ' -A solution of 290 mg of dicyclocarbodiimide in dry acetonitrile 1
0 ml was added, and the mixture was stirred at room temperature for 24 hours. Methylene chloride was added to the reaction solution to precipitate an insoluble matter, which was filtered. The filtrate was evaporated under reduced pressure, the residue was purified by silica gel column chromatography (eluting solvent: chloroform), and N-{(R) was used.
471 mg of -Nt-butoxycarbonyl-thiazolidine-4-carbonyl} -pyroglutamic acid benzylamide was obtained.

Rf value: 0.50 (developing solvent: chloroform / ethanol = 20/1) IR (KBr): ν _co 1740,1690 cm ^-1 MS: MH ⁺ -HCl, 434 NMR (CDCl ₃ ) δ: 1.30 to 1.50 (m , 9H), 2.10 ~ 2.65 (m, 3H), 2.80 ~ 3.10 (m, 2H), 3.30 ~ 3.40 (m, 1H), 4.30 ~ 4.80 (m, 5H), 5.65 ~ 5.80 (m, 1H), 6.25 -6.40 (br-s, 1H), 7.20-7.40 (m, 5H) 210 mg of the above benzylamide was dissolved in 10 ml of dry ethyl acetate, dry hydrogen chloride gas was introduced under ice cooling, and the mixture was stirred for 85 minutes. The solvent was distilled off under reduced pressure, and N-{(R)-(-)-
Thiazolidine-4-carbonyl} -pyroglutamic acid benzylamide hydrochloride (191 mg) was obtained.

IR (KBr): ν _co 1750,1690,1670 cm ^-1 MS: MH ⁺ -HCl, 334 NMR (DMSO) δ: 1.80 ~ 2.05 (m, 1H), 2.30 ~ 2.75 (m, 3H), 2.80 ~ 4.00 (M, 4H), 4.20 to 4.50 (m, 4H), 4.70 to 4.90 (m, 1H), 5.25 to 5.45 (m, 1H), 7.20 to 7.45 (m, 5H), 8.85 to 9.00 (m, 1H) Example 5 Substituting phenethylamine for benzylamine,
The following compounds were produced by the same method as in Example 4.

N-{(R)-(-)-thiazolidine-4-carbonyl} -pyroglutamic acid phenethylamide hydrochloride (Compound E) IR (KBr): ν _co 1740,1690,1660 cm ^-1 MS: MH ⁺ -HCl , 348 NMR (DMSO) δ: 1.65 to 1.90 (m, 1H), 2.20 to 2.40 (m, 1H), 2.45 to 2.80 (m, 4H), 2.85 to 4.10 (m, 4H), 4.25 to 4.45 (m, 2H), 4.60 to 4.75 (m, 1H), 5.25 to 5.40 (m, 1H), 7.15 to 7.40 (m, 5H), 8.35 to 8.55 (m, 1H) Example 6 N-{(R)-(- ) -Thiazolidine-4-carbonyl} -phenacyl pyroglutamate hydrochloride (Compound F) N-{(R) -Nt-obtained by the method described in Example 4.
Butoxycarbonyl-thiazolidine-4-carbonyl}
-792 mg of pyroglutamic acid was dissolved in 20 ml of dry N, N-dimethylformamide, 250 mg of sodium hydrogen carbonate was added, and the mixture was stirred at room temperature for 1 hour. Phenacyl bromide (500 mg) was added to the reaction solution, and the mixture was stirred at 50 ° C for 19 hours. The solvent was distilled off under reduced pressure, ethyl acetate was added, and the mixture was washed with water. After drying over anhydrous magnesium sulfate, the ethyl acetate layer was evaporated under reduced pressure, and the residue was subjected to silica gel chromatography (elution solvent:
Chloroform), N-{(R)-(-)-N
300 mg of phenacyl-t-butoxycarbonyl-thiazolidine-4-carbonyl} -pyroglutamate was obtained.

Rf value: 0.47 (developing solvent: chloroform / ethanol = 50/1) IR (KBr): ν _co 1745,1700 cm ^-1 MS: MH ⁺ , 463 NMR (CDCl ₃ ) δ: 1.39,1.47 (s, s , 9H, isomer), 2.30 ~ 3.20 (m, 5H), 3.45 ~ 3.65 (dd, 1H), 4.45 ~ 4.75 (m, 2H), 4.85 ~ 5.10 (m, 1H), 5.15 ~ 5.90 (m, 3H ), 7.50 (t, 2H), 7.63 (t, 1H), 7.88 (d, 2H) 280 mg of the above phenacyl ester was added to 10 ml of dry ethyl acetate.
Was dissolved in, and dry hydrogen chloride gas was introduced under ice cooling, and the mixture was stirred for 45 minutes. The solvent was distilled off under reduced pressure, and N-{(R)-
232 mg of (-)-thiazolidine-4-carbonyl} -phenacyl pyroglutamate hydrochloride was obtained.

IR (KBr): ν _co 1750,1695 cm ^-1 MS: MH ⁺ , 363 NMR (DMSO) δ: 2.30 ~ 2.89 (m, 3H), 3.05 ~ 3.15 (m, 1), 3.30 ~ 3.65 (m, 2H ), 4.25 ~ 4.45 (m, 2H), 4.90 ~ 5.10 (m, 1H), 5.30 ~ 5.45 (m, 1H), 5.50 ~ 5.57 (m, 2H), 7.50 ~ 7.80 (m, 3H), 7.90 ~ 8.10 (M, 2H) Example 7 N-{(L) -2-oxo-pyrrolidine-5-carbonyl} -benzyl pyroglutamate (Compound G) Benzyl pyroglutamate (5.2 g) was dissolved in dry benzene (100 ml), and the mixture was cooled with ice. 1.0 g of sodium hydride (oiliness) was added, and the mixture was stirred at room temperature for 1 hour. (L) -N in the reaction mixture
-Benzyloxycarbonyl-pyroglutamic acid N-hydroxysuccinimide ester (8.9 g) was added and the mixture was stirred overnight. After completion of the reaction, the reaction solution was washed with water, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (eluting solvent: chloroform), and 5.7 g of N-{(L) -N-benzyloxycarbonyl-2-oxo-pyrrolidine-5-carbonyl} -benzyl pyroglutamate (diastereomer) A,
2.9 g: diastereomer B, 2.9 g) was obtained.

Diastereomer AIR (KBr): ν _co 1790,1740,1710, 1700 cm ^-1 NMR (CDCl ₃ ) δ: 1.90 to 2.80 (br-s, 8H), 4.73 (dt, 1H), 5.05 to 5.30 ( m, 2H), 5.24 (dd, 2H), 5.70 to 5.80 (m, 1H), 7.30 to 7.50 (m, 10H) Diastereomer B IR (neat): ν _co 1790,1750,1710 cm ^-1 NMR ( CDCl ₃ ) δ: 1.80 to 2.90 (br-m, 8H), 4.82 (dd, 1H), 5.00 to 5.40 (m, 4H), 5.75 to 5.85 (m, 1H), 7.25 to 7.45 (m, 5H) Above Benzyl ester (diastereomer A) (2.2 g) was dissolved in ethanol (100 ml) to obtain 10% palladium-carbon (280 mg).
Was added, and the mixture was stirred under a hydrogen stream at room temperature for 6 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was evaporated under reduced pressure. The residue was recrystallized from methanol-ethyl acetate to give 600 mg of N-
{(L) -2-oxo-pyrrolidine-5-carbonyl}
-Pyroglutamic acid (diastereomer C) was obtained.

Diastereomer C Melting point: 198-203 ° C (decomposition) IR (KBr): ν _co 1740,1700,1660, 1640cm ^-1 NMR (DMSO) δ: 1.80-2.20 (m, 4H), 2.20-2.70 (m, 4H), 4.63 (dd, 1H), 5.05 (dd, 1H), 7.83 (s, 1H), 12.70 to 13.30 (m, 1H) Elemental analysis value: (as C ₁₀ H ₁₂ O ₅ N ₂ ) C% H % N% Calculated value 50.00 5.04 11.66 Measured value 47.81 5.12 11.03 Diastereomer D was produced from diastereomer B in the same manner as above.

Diastereomer D Melting point: 185 to 192 ° C (decomposition) IR (KBr): ν _co 1730,1700,1650 cm ^-1 NMR (DMSO) δ: 1.80 to 2.20 (m, 4H), 2.20 to 2.80 (m, 4H ), 4.60 (dd, 1H), 5.01 (dd, 1H), 7.91 (s, 1H), 12.80 to 13.20 (br-s, 1H) Elemental analysis value: (as C ₁₀ H ₁₂ O ₅ N ₂ ) C% H% N% Calculated value 50.00 5.04 11.66 Measured value 49.25 5.30 11.18 Above pyroglutamic acid compound (diastereomer C)
200 mg was dissolved in 20 ml of dry N, N-dimethylformamide,
34 mg of 60% sodium hydride (oil) was added under ice cooling, and the mixture was stirred at room temperature for 1 hour. Benzyl bromide 0.11 in the reaction solution
ml was added, and the mixture was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, ethyl acetate was added to the residue, washed with water, and dried over anhydrous magnesium sulfate. The ethyl acetate layer was evaporated under reduced pressure, and the residue was recrystallized from ethanol-ether to obtain 85 mg of the desired product (diastereomer E).

Diastereomer E Rf value: 0.59 (developing solvent: chloroform / ethanol = 5/1) Melting point: 33 to 36 ° C MS: MH ⁺ , 331 IR (KBr): ν _co 1735,1700 cm ^-1 NMR (CDCl ₃ ) Δ: 2.05 ~ 2.80 (m, 8H), 4.85 (dd, 1H), 5.12 (dd, 1H), 5.19 (dd, 2H), 6.34 (s, 1H), 7.30 ~ 7.50 (m, 5H) diastereo Diastereomer F was prepared from Mar D in the same manner as above.

Diastereomer F Rf value: 0.64 (developing solvent: chloroform / ethanol = 5/1) Melting point: 88 to 93 ° C IR (KBr): ν _co 1750,1710,1690 cm ^-1 MS: MH ⁺ , 331 NMR ( CDCl ₃ ) δ: 2.00 to 2.80 (m, 8H), 4.80 (dd, 1H), 5.08 (dd, 1H), 5.21 (dd, 2H), 5.79 (s, 1H), 7.30 to 7.45 (m, 5H) elemental analysis: (C ₁₇ H ₁₈ O ₅ as _{N 2) C% H% N} % calculated 61.95 5.60 8.31 Found 61.81 5.49 8.48 example 8 N - {(L) -2- oxo - pyrrolidine-5-carbonyl } -Phenethyl pyroglutamate (Compound H) N-{(L) -2-oxo-obtained by the method described in Example 7.
Pyrrolidine-5-carbonyl} -pyroglutamic acid (diastereomer D) (200 mg) was dissolved in dry N, N-dimethylformamide (15 ml), and 60% sodium hydride (oil) (34 mg) was added under ice-cooling at room temperature for 1 hour. It was stirred. Furthermore, 0.13 ml of phenethyl bromide was added, and the mixture was stirred overnight at room temperature. After completion of the reaction, the solvent was distilled off under reduced pressure, ethyl acetate was added and the mixture was washed with water, and then the ethyl acetate layer was distilled off under reduced pressure.
The residue was purified by silica gel column chromatography (eluting solvent: chloroform / ethanol = 50/1) to obtain 98 mg of the desired product.

Rf value: 0.68 (developing solvent: chloroform / ethanol = 5/1) IR (neat): ν _co 1740,1700 cm ^-1 MS: MH ⁺ , 345 NMR (CDCl ₃ ) δ: 1.90 to 2.70 (m, 8H ), 2.97 (t, 2H), 4.42 (t, 2H), 4.71 (dd, 1H), 5.03 (dd, 1H), 5.77 (s, 1H), 7.20 to 7.40 (m, 5H) Example 9 PEP inhibition Activity measurement experiment Using Z-Gly-Pro-β-NA as a substrate, the inhibitory activity against bovine brain-derived PEP was measured.

(Measurement method) 20 mM containing 10 mM EDTA and 10 mM 2-mercaptoethanol
Tris-HCl buffer (20 mM-Tris HCl Buffer, pH = 7.0)
100 μl of PEP (about 0.14 u / ml) in 0.7 ml and each concentration (0, 1
0 ^-9 to 10 ^-4 M) was added to the test compound solution (100 µl), and preincubation (Preincubat) was performed at 37 ° C for 5 minutes.
ion). Then, each concentration (5.0, 2.5, 1.25, 0.625, 0.3125 m) dissolved in 100 μl of 40% dioxane was added to this.
M) substrate was added, and incubation was again carried out at 37 ° C. for 15 minutes to allow the enzymatic reaction to proceed. Stop the reaction with 25% trichloroacetic acid, centrifuge at 3000 rpm for 10 minutes, collect 0.5 ml of the supernatant, add 0.5 ml of 0.1% nitrous acid, and after 3 minutes 0.05%. N- (1-naphthyl)
Ethylenediamine dihydrochloride ethanol solution was added. After the mixture was left at 37 ° C for 25 minutes, the absorbance at 570 nm was measured, the oxygen activity at each concentration was calculated by the following formula, and the 50% inhibitory concentration (IC ₅₀ value) was calculated from each activity value. .

Enzyme activity unit (μmol / min / ml) = ΔOD × 0.42 × dilution rate (result) Compound IC ₅₀ value Compound F 650 μM Compound G 790 μM (Geostereomer E)

Claims (1)

[Claims] 1. A general formula (In the formula, R is a 5-membered nitrogen-containing saturated heterocyclic group which may contain a sulfur atom, X is a single bond or -CO-, and Y is -O- or -NH-. , N is an integer of 1 to 2) and a pyroglutamic acid derivative and a pharmacologically acceptable salt thereof.